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PROCEEDINGS OF INTERNATIONAL CONFERENCE ON APPROPRIATE TECHNOLOGY DEVELOPMENT (ICATDev) 2015

The First International Conference on Appropriate Technology Development “Appropriate Technology for Sustainable Agriculture Development”

October 5-7, 2015 | Bandung, Indonesia

Organized by: Indonesian Institute of Sciences Center for Appropriate Technology Development

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PROCEEDINGS OF INTERNATIONAL CONFERENCE ON APPROPRIATE TECHNOLOGY DEVELOPMENT (ICATDev) 2015 First Edition: 2016 A catalogue record for this publication is available from British Library.

Disclaimer Every effort has been made in preparing this book to provide accurate and up-to-date information which is in accord with accepted standards and practice at the time of publication. Nevertheless, the authors, editors and publishers can make no warranties that the information contained herein is totally free from error. The authors, editors and publishers therefore disclaim all liability for direct or consequential damages resulting from the use of material contained in this book. Readers are strongly advised to pay careful attention to information provided by the book.

Copyright Copyright ©: 2016 by Indonesian Institute of Sciences, Center for Appropriate Technology Development.

Cover design and layout Wawan Agustina

Organized by: Indonesian Institute of Sciences Center for Appropriate Technology Development Address: K.S. Tubun Street, No. 5 Subang, West Java, Indonesia Telp. +62 260411478; +62 260412878. Fax. +62 260411239, Post code 41213

Published in March, 2016 by Sciemcee Publishing, London. LP22772, 20-22 Wenlock Road London, United Kingdom N1 7GU The proceedings with all papers are available at www.sciemcee.org.

ISBN 978-0-9935191-1-6

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PREFACE The First International Conference on Appropriate Technology Development was held on the Aston Tropicana Hotel in Bandung, Indonesia during October 5th-7th, 2016. The theme of this conference was “Appropriate Technology for Sustainable Agriculture Development”. This conference was organized by Centre for Appropriate Technology Development, Indonesian Institute of Sciences and was a part of Science and Technology Festival that held by the Deputyship of Engineering SciencesIndonesian Institute of Sciences. The aims of this conference are to develop an international network of appropriate technology researchers and practitioners committed in assisting rural communities and small and medium enterprises. Then, dissemination appropriate technology suitable for income generation through sustainable agriculture, and understanding the impact and implications of national policies for making recommendations for the extension of appropriate technology on rural income generation under the sustainable agriculture development framework. About 40 scientific participants had many fruitful discussions and exchanges that contributed to the success of the conference. Participants from some countries made the conference truly international in scope. The 29 selected papers were presented during the conference formed the heart and provided many opportunities for discussion. The papers were split almost equally between the three main conference areas, i.e. appropriate technology in agricultural technology, appropriate technology in food and feed technology, and appropriate technology in economic, environment, energy, and management. There were two plenary lectures in the conference. The First, Prof. Dr. Suwit Laohasiriwong, Professor in agricultural science Institute for Dispute Resolution, Khon Kaen University Thailand, was presented lecture entitled “Sustainable agriculture development through appropriate technology”. The Second is Dr. Ir. Akmadi Abbas, M.Eng.Sc, Vice Chairman of Indonesian Institute of Sciences, presented lecture entitled “Appropriate Technology Development in Indonesia”. These Proceedings provide the permanent record of what was presented. They indicate the state of development at the time of writing of all aspects of this important topic and will be invaluable to all workers in the appropriate technology field for that reason. Finally, it is appropriate that we record our thanks to our fellow members of the Organizing Committee and the Scientific Committee for their work in securing a substantial input of papers in encouraging participation from those.

General Chair

Wawan Agustina

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COMMITTEE Steering Committee : 1.

Dr. Ir. Yoyon Ahmudiarto, M.Sc. IPM. (Center for Appropriate Technology DevelopmentIndonesian Institute of Sciences, Indonesia)

2.

Dr. Rachmini Saparita (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia)

3.

Dr. Savitri Dyah W.I.K.R (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia)

4.

Dr. Rislima F. Sitompul (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia)

5.

Dr. Ir. Akmadi Abbas, M.Eng.Sc (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia) Dr. Pramono Nugroho (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia)

6. 7.

Ir. Doddy A. Darmajana, M.Si (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia)

Scientific Committee : 1.

Dr. Suharwadji K. Sentana (Research Center for Physics-Indonesian Institute of Sciences, Indonesia)

2.

Dr. Ainia Herminiati (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia)

3.

Prof. Alhussein Al Awaadh (Agricultural Engineering, King Saud University, Saudi Arabia)

4.

Dr. Noorul Hassan Zardari (Faculty of Civil Engineering Institute of Environmental and Water Resources Management (IPASA), Universiti Teknologi Malaysia, Malaysia)

5.

Prof. Shamim Ahmad (Agricultural Economics and Business Management Aligarh Muslim University, India)

6.

Dr. Muhammad Syarhabil Ahmad (Bioprocess Engineering, University Malaysia Perlis, Malaysia)

7.

Mohd Khairi Mohd Zambri, M.Eng (Universiti Teknikal Malaysia Melaka, Malaysia)

8. 9.

Dr. Norrizah Jaafar Sidik (Biology Universiti Teknologi MARA, Shah Alam, Selangor, Malaysia) Dr. Susan Kowalewski (Department of Business, D'Youville College, USA)

10. Dr. Haznan Abimanyu (Research Centre for Chemistrty- Indonesian Institute of Sciences, Indonesia) 11. Dr. Yanni Sudiyani (Research Centre for Chemistrty- Indonesian Institute of Sciences, Indonesia) 12. Norhazwani Abd Malek (Mechanical Engineering Universiti Tenaga Nasional Centre for Advanced Computational Engineering, Malaysia) 13. Prof. Benyamin Lakitan (Sriwijaya University, Indonesia) 14. Dr. Arie Dipareza Syafei (Environmental Engineering, Institut Teknologi Sepuluh Nopember, Indonesia) 15. Prof. Nurpilihan Bafdal (Padjajaran University, Bandung, Indonesia) 16. Prof. Dr. Ade M. Kramadibrata (Head of Indonesian Agricultural Engineerring Society, Branch of West Java, Indonesia)

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General Chair : 1. Wawan Agustina (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia) Organizing Committee: 1. Yusuf Andriana (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia) 2.

Febtri Wijayanti (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia)

3.

Moeso Andrianto (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia)

4.

Fahriansyah (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia)

5.

Seri Intan Kuala (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia)

6.

Ari Rahayuningtyas (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia)

7.

Carolina (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia)

8.

Diki Nanang Surahman (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia)

9.

Karlina Gusmarani (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia)

10. Tedy Mutaqin (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia) 11. R. Cecep Erwan Ardiyansyah (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia) 12. Cahya Edi Wahyu Anggara (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia) 13. EkoKuncoroPramono (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia) 14. Nok Afifah (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia) 15. Lia Ratnawati (Center for Appropriate Technology Development-Indonesian Institute of Sciences, Indonesia) KEYNOTE SPEAKERS 1.

Prof. Suwit Laohasiriwong, Ph.D Institute for Dispute Resolution, Khon Kaen University Thailand

2.

Dr. Ir. Akmadi Abbas, M.Eng.Sc. Vice Chairman of Indonesian Institute of Sciences

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TABLE OF CONTENT PREFACE .............................................................................................................................................. iii COMMITTEE ......................................................................................................................................... iv TABLE OF CONTENT ............................................................................................................................ v KEYNOTE PRESENTATION ................................................................................................................ 1 1.

Achieving Sustainability for Agricultural Development through Appropriate Technology Implementation: A dilemma of Thai farmers Prof. Suwit Laohasiriwong, Ph.D ...................................................................................................... 1

2.

The Development and Implementation of Appropriate Technology for Sustainable Agriculture Development Dr. Ir. Akmadi Abbas, M.Eng.Sc. ...................................................................................................... 7

AGRICULTURAL TECHNOLOGY ...................................................................................................... 15 3.

Utilization of Household Scale Tissue Culture Technique for Aloe Vera Seed Multiplication and its Irrigation Modulus Nurhaidar Rahman, Aida Wulansari, R. Ismu Tribowo, Wawan Agustina ..................................... 16

4.

Design of Pitcher System Irrigation as a Substitution of Imported Drip System Irrigation for Cultivating Horticulture and Food Crops R. Ismu Tribowo .............................................................................................................................. 21

5.

Design of Hammer Crusher to Support Small-Medium Enterprises (SMEs) In Mineral-Based Fertilizer Processing Kusno Isnugroho, David C Birawidha, Yusup Hendronursito ......................................................... 27

6.

An Appropriate Technology of Composting for Supporting Sustainable Agriculture Adi Mulyanto .................................................................................................................................... 33

7.

Runoff Harvesting as One of Appropriate Technology in Integrated Dry Land Farming Nurpilihan Bafdal, Sophia Dwiratna NP ........................................................................................ 39

8.

Pilot Scale Technology for Production Organic Biofertilizer Powder Starter to Support Sustainable Agriculture Development Nur Laili, Sarjiya Antonius, Yayuk Kartika, Dwi Agustiyani .......................................................... 43

9.

Influence of urea granulated Zeolite and Nitrification Inhibitors on Growth of Maize (Zea mays L. Var. B8) Oslan Jumadi, Ratna Dewi), Andi Takdir Makkulawu, R. Neni Iriany, Yusminah Hal), Hartono, St. Fatmah Hiola, Kazuyuki Inubushi .............................................................................. 48

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FOOD AND FEED TECHNOLOGY .................................................................................................. 53 10. Carcass Quality, Blood Profile and Organ Histopathology of Sheep Fed Organic Additive Contaning Probiotic and Micromineral Enriched Yeast Ade Erma Suryani, Lusty Istiqomah, Ahmad Sofyan, Awistaros Angger Sakti andMohammad Faiz Karimy ............................................................................................................ 54 11. Influence of Flouring Method on Characteristic of Tacca Flour: Phytochemical, Chemical and Resistant Starch Analysis Miftakhussolikhah, Dini Ariani, Tri Wiyono .................................................................................... 60 12. Making and Characterization of Carrageenan Single Edible Film Doddy A.D, Enny S, Nok A, Novita I, Qistia H.E ........................................................................... 65 13. The Development of Traditional Food-Based Military Ration Packed in Cans Kurniadi, A. Nurhikmat, M. Angwar, A. Susanto, Tri Wiyono, A. S. Praharasti ............................. 70 14. Screening of α-glucosidase Inhibitor-Producing Lactic Acid Bacteria from Ganoderma lucidum as Functional Food Candidate for Diabetic Rifa Nurhayati, Andri Frediansyah, Fitriana Rahmawati, Endah Retnaningrum and Langkah Sembiring ........................................................................................................................... 76 15. Sari Tempe Formulation from Local Soybean (Glycine max) and It’s Sensoris and Nutritional Characteristics Muhamad Kurniadi, Martina Andriani, Mukhamad Angwar, Yuniar Khasanaha, Deviy Novitasary Sukamta .......................................................................................................................... 82 16. Pineapple Peel as a Potential Source of Dietary Fiber Rima Kumalasari, Ainia Herminiati, R. Cecep Erwan Andriansyah .............................................. 89 17. Natural Antioxidant Activities of “Tanduk Rusa” Fern (Paltycerium coronarium) Ade Chandra Iwansyah, Dewi Desnilasari, Ismi S Hanifah ............................................................. 96 18. Potential of Suweg Starch HMT Modification as a Source of Resistant Starch Type III Raden Cecep Erwan Andriansyah ................................................................................................. 100 19. From Local Wisdom: Preliminary Antibacterial Activity of “Tanduk Rusa” Fern (Platycerium coronarium) Dewi Desnilasari, Ade Chandra Iwansyah, Ria Fauziah .............................................................. 105 20. New Development of Phytase Enzyme through modification of Substrate and Fermentation Technology Atit Kanti and I Made Sudiana ...................................................................................................... 109

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ECONOMY, ENERGY, ENVIRONMENT, MANAGEMENT .............................................................. 116 21. Interface between Food Security, Energy Sustainability and Water Accessibility Leuserina Garniati, Radisti Praptiwi1, Novieta Herdeani Sari, Yoyon Ahmudiarto, Jito Sugardjito, Alan Owen .................................................................................................................. 117 22. Preliminary Study of Agribusiness Development Based on Aloe vera (Case Study in Micro, Small and Medium Enterprises : Sari Kumetap Subang) Nurhaidar Rahman, Wawan Agustina, R. Ismu Tribowo, Cahya Edi W. A., R. Cecep Erwan and A. Wulansari ........................................................................................................................... 127 23. Potential Development of Arengapinnata, Merr Based On Local Knowledge (Case Study of Rejang Lebong) Eki Karsani Apriliyadi, Diki Nanang Surahman, Hendarwin M Astro .......................................... 134 24. Development of the Main Agroindustry Potential in Rejang Lebong District, Bengkulu Eki Karsani Apriliyadi, Diki Nanang Surahman, Hendarwin M Astro .......................................... 139 25. Yield Risk Assessment in Nutrient Film Technique for Pakcoy (Brassica rapa L.) Hydroponic Growing System Using FMEA and AHP Approach: A Case Study Yusuf Andriana, Eko Kuncoro Pramono, Cahya Edi Wahyu Anggara, Aidil Haryanto, Ignatius Fajar Apriyanto ................................................................................................................ 144 26. Nanotechnology Use of Activated Carbon Coal to Decreasethe Paramters of Peat Water in Jambi Muhammad Naswir......................................................................................................................... 151 27. Utilization of Lignocelluloses from Agricultural Waste as Raw Material for Producing Bioethanol Sriharti, Wawan Agustina, Takiyah Salim dan Lia Ratnawati ...................................................... 159 28. Influence of Local Wisdom to Prevent Disappearance of Cebong Lake in Sembungan Village Wonosobo District Putri Nurfahmia, Rizal Faozi Malika, Ratih Paniti Saria, Afid Nurkholisa .................................. 165 29. Sanitation Condition and Potential Recovery of Nutrients in Urban Area of Sub District Kiaracondong, Bandung City, Indonesia Neni Sintawardani, J. Tri Astuti, Dewi Nilawati, Ken Ushijima . ................................................ 169 30. The Potency of Cashew Kernels Processing Activity to Encourage Community Venture in Southwest SumbaRegency in East Nusa Tenggara Province Febtri Wijayanti, Fithria Novianti, Carolina ................................................................................. 177

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Achieving Sustainability for Agricultural Development through Appropriate Technology Implementation: A dilemma of Thai farmers Prof. Dr. Suwit Laohasiriwong, Ph.D Institute for Dispute Resolution, Khon Kaen University Thailand

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AGRICULTURAL TECHNOLOGY

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Utilization of Household Scale Tissue Culture Technique for Aloe Vera Seed Multiplication and its Irrigation Modulus Nurhaidar Rahman1, Aida Wulansari2, R. Ismu Tribowo1, Wawan Agustina1 1)

Center of Appropriate Technology Development - Indonesian Institute of Sciences Jl. K.S. Tubun No.5, Subang, West Java 41213. Telp. 0260-411478 Fax. 0260-411239 Email : [email protected] 2) Research Center of Biotechnology - Indonesian Institute of Sciences Cibinong Science Center, Jl Raya Bogor Km 46, Cibinong-Bogor , West Java 16911 Telp. 021- 8754587 Fax. 021- 8754588, Email : [email protected]

Abstract: The aloevera type that is being developed in Subang regency is from barbadensis type that contain substance for human body need, like vitamin of A, B1, B2, B6, B12, E and C. This plant purportedly can also heal for example diabetes and heart diseases. The development of aloevera agriculture to be conducted by farmers needs quite a lot of seeds. For that purpose a household scale tissue culture technology is to be applied to produce the seed acquired. The principle of tissue culture technique is multiplying the plants by using part of the vegetative plants and artificial media to be conducted in a sterile site. In general, the tissue culture is conducted at laboratory scale. By modification of equipments and materials and optimizing the environment for plants growth, the tissue culture can be conducted in smaller scale (household). The tissue culture technique can produce uniformed seeds i.e. seed’s characteristics similar to the parent plants and a lot of seed may be produced in a short time. It takes time about 1-2 months for first shoot growth whereas the conventional way takes about 1 year. It doesn’t need wide place and doesn’t depend on season, which means that it can be executed during the year. Sterilization method utilizes solution 30% Clorox shows very low level contamination (0 – 5%). In MS media that contain 1 mg/l of BAP, the amount of saplings shoot reaches more than 15 times during 1 month. The shoot's rooting has properly grown in MS media without hormone. A Greenhouse for plants acclimatization of tissue culture will need the use of net of agronet type with closeness of 70% and attached in 2 layers. The highest of irrigation modulus for aloevera plants occurred in August is as high as 0.18 liters/second/hectare with maximum of irrigation interval is as high as 31 days. Keywords: Household, Irrigation Modulus, Seed, Tissue Culture

Tissue culture is a plant multiplying technique by isolating the plant part such as leaf, shoot bud, and let grow its parts in artificial media in rich aseptic nutrition and grow regulator substance in translucent closed container until the plants part can multiply itself and regenerated become complete plants. The main principle is multiplying the plants by using the part of vegetative plants, utilizing the artificial media and conducted in a sterile site [7].

The development of aloevera agriculture cultivation by the farmers needs quite a lot of seeds. For that purpose the tissue culture technology in household scale will be applied to produce large amounts of seeds [5][11]. The aloevera can be sold in the form of plants, but also can be processed and produced to become several kinds of food, beverage, herb medicine and cosmetics [4]. The variety of aloevera that is now being developed in activity location is Barbadensis variety that contains A, B1, B2, B6, B12, E, and C vitamins. This plant purportedly can also heal diabetes and heart disease [3][5].

Method of tissue culture is developed to help multiply plants, especially for the plants which are difficult to be breed generatively. The seed that come up from the tissue culture have some excellences, for example: have characteristics similar to the parent plants, can be multiplied in big number and do not require wide place, able to produce number of seeds within short time, the seed is healthy and its quality are more guaranteed, the seed growing speed is quicker compared to the conventional multiplying [2][7].

Modulus of irrigation is to denote water requirement for plants viability (in this case aloevera) but does not include the efficiency factor due to water waste in the irrigation system such as evaporation and percolation. Modulus of irrigation for a plant will differ from other plants, and so goes the same with the kind of soil media that grow plants especially soil texture [9]. A farm land used to cultivate several kinds of plants will need handling of water irrigation management to fulfill the amount of water required for growth of several kinds of cultivated plants [6].

1. Introduction 1.1 Backgrounds

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1.2 The Purposes Multiplying the availability of aloevera seed by using the technique of household scale tissue culture and counting the modulus of irrigation requirement during its cultivation. 2. Methodology Tissue culture: the materials to be used include materials for media making, for explants sterilization (the tissue that will be cultivated), for planting (inoculation) and for acclimatization. Whereas the equipment to be used includes the equipments for making of tissue culture media, explants preparation (initiation), planting (inoculation), incubation and acclimatization [3][7]. Modulus of Irrigation: analysis of irrigation water requirement at plants level (irrigation modulus) consist of crops calendar determination, get data of climatology, get level of water requirement for plants, knowing naturally water supply, and get level of irrigation water requirement which is expressed in term of irrigation modulus with unit of liter/second/hectare. Analysis of irrigation water interval (which is continuance from analysis of irrigation modulus that cannot be dissociated) need calculation of average depletion rate of ground water content and calculate totalize availability of soil water content that is readily used [8][9]. Activity Location :The activity is conducted in The Center of Appropriate Technology Development Indonesian Institute of Sciences (Pusbang TTGLIPI) Subang and in The Medium Small Enterprise (UKM) Sari Kumetap which address is at Jalan Cagak, View Garden Residential Block D24 No.11 Palasari village, Sub district of Ciater, district of Subang, West Java Province which is around 20 km from Subang city on the way to Bandung. 3. Result and Discussion

Media of tissue culture which is used is the media with complete nutrition, to do so the explants can grow properly become completely plants. The media which are used must be in a state of sterile, free from microorganism either bacterium or mold. If the media is not sterile, then microbes will grow so it will bother the explants growth. The step of media sterilization uses autoclave or high pressured pan at high temperature will kill all bacterium and mold that possible exist in the media. The sterilization process not only for the grow media, but also for all of the equipments which are used in explants cultivation. The sterilization of plants tool like pinset, knife hilt and petri can be conducted along with the growth media sterilization. Sterile distilled water also must be prepared for rinsing at the moment of explants sterilization (Figure 1).

Figure 1: Growth media sterilization and plants equipments. (Source: Imelda et.al. Research Center of Biotechnology - Indonesian Institute of Sciences) A. Autoclave / pan B. Inner Autoclave C. Grow media and plants equipments are ready to be sterilized with pressure as high as 1 atmosphere and temperature as high as 121oC. 3.1.2 Explants Sterilization Phases

3.1 Tissue Culture The technique of tissue culture is a technique to multiply plants in vegetative way which needs an aseptic condition. It means that from grow media which is used till plants planting and its growth are conducted at microorganism free condition. The activity step conducted include making growth media and its sterilization, prepare explants in the kind of aloevera saplings, explants planting at media and plantlet acclimatization at soil media [3][7]. 3.1.1 Media Making Sterilization

conducted related to process of nutrition absorption and media condensation. The required range of optimum pH is + 5.8 for almost all plants types. This media is then poured into culture bottle, closed and sterilized using autoclave or pressured pan.

and

Planting

Tool

Grow media which is commonly used consist of macro and micro substances, amino acid, vitamin, sugar and grow hormone with certain comparison and condenser in the form of seaweed. The measurement of pH media is quite important to be

The explants which are used are the aloevera saplings that have an age of 2-3 months and have shoots height between 10-15 cm (Figure. 2A and 2B). The explants which are selected must be healthy saplings and have a nice growth. Hereinafter the saplings are cleared from dirt and soaked and shaken in liquid soap solution during 10 -15 minutes. Outmost leafs are eliminated. After soaking with liquid soap solution, the shoots are washed with flowing water and soaked and shaken in 30% Clorox solution for 45 minute. The step is then conducted in laminar flow. The shoots which are soaked in solution Clorox are picked out, and then cleaned 3 times with sterile distilled water. Hereinafter the shoots are cut till height of 2-3 cm, and planted in MS media without hormone (Figure 2C).

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The observation to the level of contamination of explants from sterilization, indicate that the sterilization method which is used has been optimal. Percentage of contaminable explants are between 0 – 5%.

attached 2 layers. The distance between layers is 0 up to 1 meter high. The effect of the distance range between layer and the usage of the net with 70% of closeness to the growth of tissue culture plants has not been observed. The usage of the net applied depends on the experience in the development activity of horticulture plants in the Greenhouse in The Center for Appropriate Technology Development - Indonesian Institute of Sciences in Subang [6].

Figure : 2. The aloevera explants which are used for tissue culture (Source: Imelda et.al. Research Center of Biotechnology - Indonesian Institute of Sciences). A. Aloevera parent plants that have produced saplings B. Aloevera saplings shoot which are used as explants C. Explants which are already sterilized and planted in MS media. 3.1.3 Growth Phase and Shoot Multiplying After sterilization phase, it is continued with observation to shoot growth in vitro planted. At previous research, it has been obtained the optimal media for growth and multiplication of shoot in vitro [2]. In MS media that contain 1 mg/l of BAP, the amount of the saplings shoot rattan can reach more than 15 times during 1 month. The shoot growth made improvement from first week plant till sixth week. This condition indicates that the hormone and nutrition application in plant media have already met the plants need (Figure 3). The shoot's rooting has properly grown at MS media without hormone.

Figure 3: Graph of relation between shoot numbers against plant age.(Source: Imelda et.al. Research Center of Biotechnology - Indonesian Institute of Sciences).

3.2 Acclimatization Acclimatization is an important phase, because at this phase plants (plantlet) will be adapted to live in the field, so it can become normal plants. At this phase the correctness is needed, because this phase is critical phase and often caused plantlet death. Plantlet from tissue culture will adapt either through morphology or physiology to be able to live in the field, need correctness and good knowledge to succeed. The level of plants adaptation to environment outside the culture bottle is weak. Plantlet death in general caused by high respiration of the plantlet that causative to wilt plantlet and die [5]. Greenhouseat figure 4 is needed for acclimatization of plants from tissue culture before planted at plants farm. The roof of the Greenhouse uses net from agronet type with 70% of closeness that

Figure 4: Greenhouse side view (Source: Tribowo, Pusbang TTG-LIPI)

Figure 5: Drips irrigation system (Source: Tribowo, Pusbang TTG-LIPI)

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For plants irrigation in the Greenhouse (acclimatization) can use drips irrigation system as can be seen at Fig. 5. If plants go on living in the Greenhouse after a period of acclimatization, it can be continued its cultivation in hydroponic way by utilizing drips irrigation system [8].

from aloevera plants can be seen in Table 3. The highest modulus of irrigation of aloevera plants occurred in August is as high as 0.18 liter/ second/ hectare. If the irrigation modulus is obtained, then the calculation of irrigation interval must be made by assessing the average of depletion rate of soil water content and then to be calculated to total readily availability of soil water content to be used. For silty clay loam texture, its water content level at wilting point is 18% and its water content level at field capacity is 34%. With assumption for the upper layer soil (0 s/d 15 cm) has 18% of water content at wilting point and 34% of water content at field capacity. At depth of 15 s/d 75 cm where plants root is still found, 16% of water content at wilting point is taken and 30% of water content at field capacity is taken [1]. To do so the level of soil water content at every deepness in the range of 15 - 25 35 cm are assumed to be as seen in Table 4.

3.3 Modulus of Irrigation after Acclimatization To make sure that the plants grow optimal, it also must be supported by optimal water availability that can be absorbed by plants through plants roots. Therefore, the analysis/calculation of crop water requirement or what is often called irrigation modulus must be conducted [9] [10]. Based on the situation of the precipitation and evapotranspiration on the farm land to be cultivated, a crop calendar is made to be synchronized with the crop pattern of the plants that will be planted (Table 1). By looking at the crop calendar and number of plants factor (Kc) (Table 2), and also considering the potential evapotranspiration data, precipitation and potential effective from precipitation itself, the crop water requirement will be able to be determined (irrigation modulus). Hereinafter irrigation modulus

The maximum irrigation interval (ni max) from Aloevera plants can be seen in Table 6 that is as high as 31 day that occurred on August

Table 1. Design of aloevera crop calendar Area fraction

0% 100 %

No Tis sue cult ure

Dec Seed cult ivat ion

Jan

Feb

Ma

Ap

Ma

Jun

Jul Early har vest

up till 3 years Harvest every week up till 3 years old of plants age

Table 2. Crop Calendar and Crop Factor Number

Table 3. Modulus of Irrigation (qo) of Aloevera Plants Period No Dec Jan Feb Ma Ap qo l/d/ha

Ma -

Jun -

Jul -

Aug .18

Sep .07

Oct -

Table 4. The amount of soil water content in accordance with its depth Soil profile cm 0 – 15 15 – 40 40 – 75 Explanation:

OWP V%

OFC V%

AM V%

18 16 16

34 30 30

16 14 14

OWP : moisture content at wilting point, AM : available moisture. Si : initial water supply Source: Developed from field observation result

AM mm

Oi V%

Si mm

24 18 24 35 24 15 49 30 108 39 OFC: moisture content at field capacity, Oi : initial water content

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Table 6. Maximum Irrigation interval of aloevera plants Period ni max. Aloevera

day

Jan -

Feb -

Ma -

4. Conclusion Using the technique of tissue culture uniformed seed can be produced i.e. seed‟s characteristics similar to the parent plant. The seed amount that is produced is many in a short time, which is about 1 – 2 months for first shoot growth compared to the conventional way that takes about 1 year. It does not need wide place and does not depend on season, so it can be executed during the year. Sterilization method utilizes solution 30% Clorox shows very low level contamination (0 – 5%). In the MS media containing 1 mg/l of BAP, the amount of the saplings shoot can reach more than 15 times during 1 month. The shoot growth makes improvement from first week plant till sixth week. This condition indicates that the hormone and nutrition application in plant media have already met to the plants need. The shoot's rooting has properly grown at MS media without hormone. The roof of the Greenhouse for plants acclimatization of tissue culture uses net of agronet type with closeness of 70% and attached in 2 layers. The highest of irrigation modulus for aloevera plants occurred in August is as high as 0.18 liters/second/hectare with maximum of irrigation interval is as high as 31 days. Acknowledgment We are very grateful to The Head and Staff of The Center of Appropriate Technology Development Indonesian Institute of Sciences (PPTTG-LIPI) in Subang for their support during research and development activities. We also appreciate very much all colleagues of researcher staff like Angga, Cecep, Wawan and technician colleagues like Herwanto, Sukwati, Jaja, Dedi and many others for all their cooperation and assistance. References [1]. Dijkerman, J.C., 1981, Field Description, Morphology and Sampling of Soils, Agricultural -University, Wageningen, The Netherland, 4/4 – 4/5. [2]. Imelda, M., A. Wulansari, L. Sari, F. Erlyandari, 2006, Peningkatan Kadar Aloin Lidah Buaya melalui Embriogenesis dan Mutagenesis, Laporan Teknik Kegiatan Penelitian Pusat Penelitian Bioteknologi LIPI. Cibinong, hal: 31-40. [3]. Rahman, N. dan A. Rahayuningtyas, 2013, Penerapan Teknik Kultur Jaringan Dalam Rangka Penyediaan Bibit Singkong Jenis Darul Hidayah Dalam Upaya Peningkatan Mutu Produk Olahan Singkong, Prosiding Seminar

Ap -

Ma -

Jun -

Jul -

Au 31

Sep 84

Oc -

Nasional dan Workshop Peningkatan Pemanfaatan Inovasi dalam Menanggulangi Kemiskinan, Bandung, 30 September - 1 Oktober 2013. [4]. Rahman, N., dkk., 2014, Pengembangan Usaha Agribisnis Berbasis Lidah Buaya di Usaha Kecil Mikro Sari Kumetap di Kabupaten Subang, Proposal kegiatan penerapan ilmu pengetahuan dan Teknologi di daerah (Iptekda) lipi Tahun 2015. Subang, 34 pp. [5]. Rahman, N. dan A. Rahayuningtyas, 2014, Upaya Pengenalan Teknik Kultur Jaringan dalam Pembibitan Singkong Skala Rumah Tangga (Studi Kasus : Petani dan Ibu Rumah Tangga di Lingkungan Pusbang TTG), Prosiding Konferensi & Seminar Nasional Teknologi Tepat Guna Tahun 2014. Peranan Teknologi Tepat Guna untuk Meningkatkan Daya Saing Bangsa, Pusat Pengembangan Teknologi Tepat Guna- Lembaga Ilmu Pengetahuan Indonesia, Subang, hal. 397- 404. [6]. Rahman, N. dkk., 2014, Urban Farming dan Tinjauan Teknologi Managemen Air dan Tanah Sebagai Salah Satu Pendukungnya, Prosiding Konferensi & Seminar Nasional Teknologi Tepat Guna 2014, P2TTG-LIPI – PNPM – PISEW, Bandung, hal. 420 – 433. [7]. Sriyanti dan Daisy, P., 1994, Teknik Kultur Jaringan, Kanisius, Yogyakarta. [8]. Tribowo, R.I., 1998, Panduan Teknis Perancangan Irigasi Sistem Tetes/Drip, Balai Pengembangan Teknologi Tepat Guna, Puslitbang Fisika Terapan, Lembaga Ilmu Pengetahuan Indonesia, Subang, 28 pp. [9]. Tribowo, R.I. dan D.A. Darmayana, 2002, Analisis modulus dan interval irigasi dalam perancangan sistem irigasi bertekanan untuk tanaman hortikultura, Balai Pengembangan TTG – LIPI, Subang, 13 pp. [10]. Tribowo R.I. dan Carolina, 2000, Analisis perancangan sistem irigasi kendi untuk tanaman lidah buaya sebagai tanaman utama dan tanaman hortikultura sebagai tanaman sela, Balai Pengembangan TTG – LIPI, Subang, 15 pp. [11]. Sandra, W, 2012, Salah Persepsi Tentang Kultur Jaringan Tanaman Skala Rumah. www.facebook.com/notes/edhi-sandra/salahpersepsi-tentang-kultur-jaringantanaman-skalarumah-tangga (29 Oktober 2013).

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Design of Pitcher System Irrigation as a Substitution of Imported Drip System Irrigation for Cultivating Horticulture and Food Crops R. Ismu Tribowo Center for Appropriate Technology Development – Indonesian Institute of Sciences K.S.Tubun No.5 Subang-West Java-Indonesia, 41213 Tlp. 0260-411478, Fax. 0260-411239 e-mail : [email protected]

Abstract : Irrigation of drip system has been pertained rareness its use in Indonesia. To lessen dependable at imported equipments then a substitution of system irrigation must be made for example is pitcher system irrigation with its technical capability that not fails against imported equipment. The highest modulus of irrigation for pepper plants is occurred on August, where required water as high as 0.32 litres/second/hectare. With 95% of efficiency, then the water irrigation requirement that required from the pitcher is 0.34 litres/second/hectare. Water source is taken from the artesian drill well. The area of the cultivating farm is 2 hectares that consist of 20 units of 1000 m2 each of farm field. From 1000 m2 of farm field, 36 of beds were made with high 40 cm, wide 100 cm and length 2000 cm. Distance between beds is 40 cm. Every bed is attached with one lateral pipe. Along the lateral pipe existed distribution hose pipe to fill water into the pitcher. The average of the pitcher's water stream flow is 0.6 litres/hour. The pitcher is put down in the ground limited to the pitcher neck and the water level in the pitcher relative fixed during irrigation operational that is 1 up till 2 cm below surface of the pitcher neck by utilizing mariote tube principle. The irrigation interval is once in a day that is at morning or evening. The maximum of the duration time of the irrigation water delivery for pepper plant is 103 minute, tomato 103 minute, grape 78 minute, banana 75 minute, pineapple 10 minute, paddy 90 minute and maize 71 minute. The mariote tube can use fiberglass container with its volume of 5.000 litres. For every 1.000 m2 of farm field can use 1 container unit or more, depend on the cultivated plants. The investment cost of the pitcher system irrigation can save its cost more than 40% compared to the imported drip system irrigation. Keywords : design, food crops, horticulture, import substitution, pitcher system irrigation 1. Introduction 1.1 Background Irrigation of drip system has been pertained rareness in its use in Indonesia. This rare because of its equipments from irrigation of drip system it self is not available, with other word if we want to use its drip system, then the equipments must be imported (Figure. 1). In other hand its investment also relatively quite expensive. To lessen dependable at imported equipments then must made a substitution of system irrigation for example is pitcher system irrigation (Figure 2) with its technical capability that not fail against imported equipment. Like at drip system irrigation, pitcher system irrigation can be used at horticulture plants like fruits and vegetable, can be also used for cultivating of food plants like maize and dry land paddy.

Figure 1. Imported dripper along with distribution hosepipe

Figure 2. Two (2) units of examples pitcher for plants irrigation Conceptually, Perhimpunan Meteorologi Pertanian Indonesia (PERHIMPI), Perhimpunan Agronomi Indonesia (PERAGI) and Perhimpunan Ekonomi Pertanian Indonesia (PERHEPI) (1996) has publicized “Gerakan Hemat Air" (GHA) or "Water Saving Movement” as the effort of water resources utilization efficiently [1] especially in its connected with the anticipation and prevention of dryness that continued with Pencanangan Gerakan Hemat Energy dan Air(Announcement of Energy and water saving movement) by President of R.I. Susilo Bambang Yudoyono in 10 Augusts 2008 at the cross square of South Monas, North of Jakarta [7].

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Proceeding of International Conference, 2015 “Water Saving” in Water Saving Movement interpreted as the effort to avoid loses at all phase of water flows on the surface of the earth. In consequence, GHA are addressed to improve water availability, delivering and usage efficiency, and depress losing and its usage wasting [2]. Some technologies and irrigation management with highly efficient in utilization of existing water sources for especially agriculture purpose which during the time is developed by Center of Appropriate Technology Development - Indonesian Institute of Sciences, for example is sprinkler irrigation, drip, and pitcher irrigation with its mariote tube. Alternative to anticipate that above and at the same time also in order to pro active joining the save water movement activity that announced by President R.I. since 1994, in agriculture area is its utilization of the highly efficient irrigating technologies in irrigation water usage [3]. The technology of pitcher system Irrigation is known have level of significant highly efficient water usage compared to opened channel irrigation or gravitation is alternative that can be implemented. For the application alternative of appropriate of technology for economical irrigation water purpose, required data of farm climate and plants agronomy that cultivated, soil structure and texture data, quality and quantity of water source and the specification of the equipment that used. Management of operational and maintenance of irrigation system that used is a part of the activity of its irrigation system for economical irrigation achievement and also efficient either in arrangement of irrigation time or in utilization of limited water source.

the tube has fixed pressure and although the water level in tube is lowered, the surface water level in every pitcher is always constant and equal [3]. The illustration of the work principle of the Mariote Tube for pitcher system irrigation can be seen at Figure 5.

Figure 3. Irrigation pitcher

1.2 Purposes Designing the pitcher system irrigation with local materials as a substitution of the imported drip system irrigation by using some of its imported drip system irrigation designed as references.

2. Methodology

Figure 4. Form and specification of irrigation pitcher (Source : Setiawan B.I.)

Design of pitcher system irrigation covers crops calendar and crops pattern design, calculation of crop water requirement at plants level (modulus of irrigation), calculation of maximum of irrigation interval, calculation of maximum of average water stream flow of every pitcher, and making of time schedule of irrigation rotation when necessary. Materials of the pitcher are designed specially by mixing clay, sand and sawdust. The best Mixture is that produce pitcher's permeability equal to soil permeability. Form and specification of irrigation pitcher can be seen at Figure. 3 and 4. The work principle of the Mariote Tube is used for giving irrigation water of pitcher system. The pitcher is immersed in the ground till limited to the pitcher's neck. Each pitcher is connected by using plastic pipe to deliver water from the tube. Water that flow from

Figure 5. Mariote Vacuum Tube for pitcher system irrigation

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Activity Location Design Study is conducted for farm field of ex UPT - Balai Bahan Olahan Kimia - Indonesian Institute of Sciences, Purwotani Village, Sub District of Tanjung Bintang, District of South of Lampung, Province of Lampung. At farm for cultivating of horticulture plants for the area of 2 hectares has farm inclination from direction of South to North about 2,7 %, whereas from direction of West to East about1,5 %. Methodology 2.1 Irrigation Water Requirement at Plant Level (Modulus of Irrigation). The calculation of modulus of irrigation [4] use equation: qo fld = 100/ea x qo ltr/sec/ha (1) qo = IR (mm/day) x 0.116 (ltr/sec/ha) (2) (mm/day) IR = WR – WS (3) WS = R.Eff.pot. x a x t (4) R.eff.pot. = R x Pot.Eff. (5) WR = (Etm x a1 x t) + (S x a2) (6) Etm = Kc x Eto (7)

Explanation of symbols qo : modulus of irrigation qo fld : water requirement at the emitter level ea : field irrigation efficiency WS : natural water supply

R.Eff.pot. : Pot. Eff. : R a t WR Etm a1 a2 S Kc Eto

potential effective rainfall percent amount of rainfall that absorbed by the soil : average monthly rainfall : area fraction : time fraction : water requirement : Maximum evapotranspiration : area fraction to Etm : fraction area to irrigation water requirement : water supply : crop factor : evapotranspiration

2.1.1 Determination of Pattern and Calendar of Crops

From crops calendar (Table 1.) and number of crops factor (Kc) can be known the water requirement that needed [5]. Pattern and calendar of crop for pepper plants is used due to pepper is the crop of vegetable horticulture with high irrigation modulus. 2.1.2 Irrigation Modulus (qo)

Where this is the amount of irrigation water requirement at plants level with unit of litre/second/hectare like at Table 2.

Table 1. Crop calendar for pepper included period fraction ( a: area , t: time )

Explanation : Kc : crops factor; Spec.req: water requirement needed

Table 2. Irrigation modulus and water requirement at the pitcher level

Explanation :

 ea

: Assumption of estimation of pitche system irrigation water at level of plants farm that due to lose water because of evaporation during water delivery, and lose water because of unequally water distribution in root zone.  qo fld. : irrigation water requirement at pitcher level

The highest modulus of irrigation for pepper plants is occurred on August, where required water as high as 0.32 litres/second/hectare or 2.8 mm/day. With 95% of efficiency, then the water irrigation requirement that required from the pitcher is 0.34 litres/second/hectare or 2.95 mm/day. 2.2 Maximum Interval of Irrigation The calculation of maximum interval of irrigation [4] use equation: ni max. = TRAM/ qd TRAM = AM x P qd = Etm - R eff.pot. + qp - qc Explanation of symbols ni max.

:

(8) (9) (10)

maximum interval of irrigation

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TRAM AM P

: : :

qd qp qc

: : :

totally soil water content availability soil water content that can be used depletion fraction of soil water content average of depletion rate percolation rate capilary rate

The determination of the optimal irrigation interval is very necessary especially at peak dry season where the debit of irrigation water that given reach its peak as well. The result of the irrigation interval calculation of every month period presented at Table 3. From the number of the irrigation interval got that on August has occurred the shortest interval that is 16 day. Accordingly the design of the irrigation system is based on 16 day irrigation interval or smaller. Table 3. Maximum interval of irrigation

2.3 Maximum of Duration Time of Pitcher Irrigation The calculation of maximum the duration time irrigation of every pitcher require data from irrigation modulus and pitcher specification it self. Moreover require also map of its irrigation scheme layout. From the layout of imported drip irrigation scheme (Figure 6), at the same farm the pitcher irrigation scheme layout is made and it can be seen at Figure 7. Water source is taken from the artesian drill well. The area of the cultivating farm is 2 hectares that consist of 20 units of 1000 m2 each of farm field (Figure 8) From 1000 m2 of farm field, 36 of beds were made with high 40 cm, wide 100 cm and length 2000 cm. Distance between beds is 40 cm. Every bed is attached with one lateral pipe. For weed control and evaporation reduction from ground surface, then every bed is given black silver plastic mulch cover. Along the lateral pipe existed distribution hose pipe to fill water into the pitcher. In the design, pitcher with average of pitcher's water stream flow of 0.6 litres/hour (the testing result in the laboratory of Water and Soil Management of Center of Appropriate Technology Development Indonesian Institute of Sciences in Subang) is used, where the pitcher is put down in the ground limited to the pitcher neck and the water level in the pitcher relative fixed during irrigation operational that is 1 up till 2 cm below surface of the pitcher neck by utilizing mariote tube principle.

Figure. 6 Map of drip system irrigation scheme layout for area of 20,000 m2. The highest modulus of irrigation for pepper plants is occurred on August, where required water as high as 0.32 litres/second/hectare or 2.8 mm/ day (Table 2.). With assumption of water utilization efficiency as high as 95%, then the irrigation water requirement that released by the pitcher is 2.95 mm/day [6]. From Table 3. (irrigation interval) it has been designed that the irrigation interval is once in a day that is at morning or evening. Accordingly the depth of the irrigation water that is given for every operational in such condition is 2.95 mm.

Figure 7. Map of pitcher system irrigation scheme layout for area of 20,000 m2 With assumption of soil wetness percentage due to water infiltration by pitcher as emitter is 100% and the area of wet soil is 0.35 m2/pitcher (distance plants 0.5 m x 0.7 m, one pitcher is put down between two plants, so it's can be told that one pitcher unit serves one plants) then the water volume at wet soil because of infiltration of every pitcher is: 0.35 m2 x 2.95 mms = 1.03 litres. Maximum of the duration time of irrigation water delivery (if the average of water seepage rate of every pitcher as the emitter is 0.6 litres/hour) for pepper plant is 1.03 litres: 0.6 litres/hour = 1 hour 43 minute. The duration time of irrigation water delivery for other time and plants can be seen at Table 4.

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Table 4. The duration time of irrigation water delivery at pitcher level

Note:must be considered also the duration of filling up of water to the container and the mariote vacuum tube

2.4 Pitcher Irrigation for Food Crops This system pitcher Irrigation can be also used to irrigate food crops cultivation such as maize and dry land paddy. The calculation of water requirement and the duration time of irrigation water delivery can refer to the calculation for pepper cultivation previously. The Irrigation Modulus and the duration time of irrigation water delivery through the pitcher's seepage for maize and dry land paddy can be seen at Table 5. Table 5. Irrigation Modulus (qo) and the duration time of irrigation water delivery through the pitcher's seepage for maize and dry land paddy.

attached with one lateral pipe. For weed control and evaporation reduction from ground surface, then every bed is given black silver plastic mulch cover. Along the lateral pipe existed distribution hose pipe to fill water into the pitcher. In the design, pitcher with average of pitcher's water stream flow of 0.6 litres/hour, where the pitcher is put down in the ground limited to the pitcher neck and the water level in the pitcher relative fixed during irrigation operational that is 1 up till 2 cm below surface of the pitcher neck by utilizing mariote tube principle. The irrigation interval is once in a day that is at morning or evening. Accordingly the depth of the irrigation water that is given for every operational in such condition is 2.95 mm. Maximum of the duration time of irrigation water delivery (if the average of water seepage rate of every pitcher as the emitter is 0.6 litres/hour) for pepper plant is 103 minute, tomato 103 minute, grape 78 minute, banana 75 minute, pineapple 10 minute, paddy 90 minute and maize 71 minute.

2.5 Capacity of Mariote Tube Volume The mariote tube can use fiberglass container with its volume of 5,000 litres. For every farm of 1.000 m2 can use 1 container unit or more, depend on the cultivated crops. 2.6 Analysis of Engineering Economy Based on references of [4] in Break Even Point Analysis in Design of Drip System Irrigation for Farm Field of Multicrops of Horticulture Crops and [6] in Analysis of Engineering Economy of Utilization of Pitcher System Irrigation, in rough calculation, investment cost of pitcher system irrigation can save more than 40% compared to imported drip system irrigation. More detail calculation can be conducted at next step.

3. Conclusion and Suggestion The highest modulus of irrigation for pepper plants is occurred on August, where required water as high as 0.32 litres/second/hectare or 2.8 mm/day. With 95% of efficiency, then the water irrigation requirement that required from the pitcher is 0.34 litres/second/hectare or 2.95 mm/day.Water source is taken from the artesian drill well. The area of the cultivating farm is 2 hectares that consist of 20 units of 1000 m2 each of farm field. From 1000 m2 of farm field, 36 of beds were made with high 40 cm, wide 100 cm and length 2000 cm. Distance between beds is 40 cm. Every bed is

Figure 8. Pitcher system irrigation layout for farm field of 1000 m2 (not on scale) The irrigation interval is once in a day that is at morning or evening. Accordingly the depth of the irrigation water that is given for every operational in such condition is 2.95 mm. Maximum of the duration time of irrigation water delivery (if the average of water seepage rate of every pitcher as the emitter is 0.6 litres/hour) for pepper plant is 103 minute, tomato 103 minute, grape 78 minute, banana 75 minute, pineapple 10 minute, paddy 90 minute and maize 71 minute. The mariote tube can use fiberglass container with its volume of 5,000 litres. For every farm of 1.000 m2 can use 1 container unit or more, depend on the cultivated crops. Investment cost of pitcher system

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irrigation can save more than 40% compared to imported drip system irrigation. More detail calculation can be conducted at next step. More detail calculation is suggested for irrigation time for area of 20.000 m2 of farm field. The calculation depends on the cultivated crops. And so do for making rotation schedule of water delivery also depend on the cultivated crops.

[7]. http://www.setneg.go.id/1 Februari 2012, “Upacara Pencanangan Gerakan Hemat Energi dan Air” oleh Presiden R.I. Susilo Bambang Yudoyono, di Lapangan Silang Monas, 10 Agustus 2008. May 14, 2013.

Acknowledgment Thank you we say to The Head of Center of Appropriate Technology Development - Indonesian Institute of Sciences and staff to the all its amenity and aid and also friends like Elok W.H., Taufik, Aden and friends from Bogor Agriculture Institute like Budi I.S., and Saleh, also to those all that cannot we mention one by one to the all its good aid and cooperation so this activity can be accomplished properly.

References [1]. Anonim, “Pemantapan Gerakan Hemat Air untuk Mengoptimalkan Pemanfaatan Sumber Daya Air”, Seminar Nasional Gerakan Hemat Air, Jakarta, 1996. [2]. Baharsjah, J.S., “Optimasi Permanfaatan Air Irigasi di Tingkat Usaha Tani Sebagai Implementasi Gerakan Hemat Air”, Makalah Seminar Nasional Himpunan Ahli Teknik Tanah dan Air kerja sama dengan Komite Nasional Indonesia untuk ICID, Bekasi, 1997, 26 pp. [3]. Setiawan, B. I. , “Sistem Irigasi Kendi Untuk Tanaman Sayuran Di Daerah Kering”, Laporan Riset Unggulan Terpadu IV, Fakultas Teknologi Pertanian, Institut Pertanian Bogor, Bogor, 1998, 125 pp. [4]. Tribowo R.I., “Perancangan Sistem Irigasi Tetes Ulir Plastik sebagai Substitusi Sistem Tetes Impor untuk Budidaya Tanaman Hortikultura dan Pangan”, Prosiding Seminar Nasional Ilmu Pengetahuan Teknik “Teknologi untuk Mendukung Pembangunan Nasional”, Pusat Penelitian Elektronika dan Telekomunikasi – Lembaga Ilmu Pengetahuan Indonesia, Bandung, 2012, Hal. 116 – 122. [5]. Doorenbos, J., et.al., “Crop Water Requirements”, FAO Irrigation and Design Paper No. 24, Rome, 1984, 144 pp. [6]. Tribowo R.I., “Analisis Pemanfaatan Kendi Sebagai Emiter Irigasi Budidaya Tanaman Jagung di Lahan Kering”, Makalah pada Seminar Nasional Peningkatan Pemanfaatan Inovasi dalam Menanggulangi Kemiskinan pada 16 Agustus 2013, Lembaga Ilmu Pengetahuan Indonesia, Bandung. 2013, Hal. 55 - 63. 1st International Conference on Appropriate Technology Development 2015, Bandung, October 5th-7th, 2015 | 26

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Design of Hammer Crusher to Support Small-Medium Enterprises (SMEs) In Mineral-Based Fertilizer Processing Kusno Isnugroho, David C Birawidha, Yusup Hendronursito* Technical Implementation Unit of Mineral Processing, Indonesian Institute of Sciences, Lampung,35364,Indonesia Tel.: +62-721-350052; fax: +62-721-350054. E-mail address: [email protected]; [email protected]; [email protected] Abstract : Dolomite and zeolite are industrial minerals that can be used as an alternative to mineral-based fertilizers. The existence of zeolite and dolomiteare scattered in various regions in Indonesia. This condition is an opportunity for small and medium industries (SMEs) to undertake theprocessing of these minerals as raw materials of mineral-based fertilizers. SMEs can perform the processing of these minerals in the preparationstage of the crushing the materials. This research was conducted to design and preliminary test of hammer crusher to support SMEs in mineralbasedfertilizer processing. Hammer crusher used is a double-shaft hammer crusher. The theoretical capacity is 1,000 kg/hour of raw material(dolomite and zeolite). The hammer crusher real capacity was 825-900 kg / hour of raw materials at 2,900 rpm, and 685-750 kg / hour of raw at1,450 rpm. At 1,450 and 2,900 rpm, obtained the efficiency of hammer crusher to process dolomite was 82.11% and 88.54%, for zeoliteefficiency was 88.79% and 94.81%. Keywords:crusher , design ,double -shaft, dolomite, hammer, zeolite. 1. Introduction

μ

Fertilization is one way that can be used to increase agricultural output. So far, there are two types of fertilizers, which are a single fertilizer and compound fertilizer. Single fertilizer consists of one type of nutrients, such as urea (nutrient nitrogen), TSP (nutrient phosphorus) and KCl containing nutrient potassium. Compound fertilizer is a fertilizer containing more than one kind of nutrients, such as NPK contains nitrogen, phosphorus and potassium. Compound fertilizers can be obtained from mineral resources / minerals industry so called alternative mineral-based fertilizers. The advantages of mineral-based fertilizers are the mineral raw materials that are easily obtainable, the manufacturing process is easy, and the cost of production is much cheaper compared to the manufacture of chemical-based fertilizers. With some of the advantages, this fertilizer can be used as an alternative, especially when fertilizer demand continues to increase (Adang,2011).

T1 T2 M V S A P T fs d

Nomenclature Q P V t n1 n2 D1 D2 L C β R r α1,2

capcity, ton/hour bulk density, kg/cm3 volume, m3 time, hour speed of the driver, rpm speed of the follower, rpm diameter of driver, m diameter of the follower, m total length of the belt, m distance between centre of the two pulley, m belt contact angle, radian radius of the large pulley,mm radius of the smaller pulley, mm angle of wrap for the pulley, deg

coefficient of friction between the belt and the pulley the tension in the tight side of belt, N the tension in the slack side of belt, N mass per unit length of belt linear velocity of belt the maximum permissible belt stress, MN/m2 area of belt electropower, watt torque,Nm shear stress, Nm/mm2 axle diameter, mm

Dolomite and zeolite are industrial minerals that can be used as an alternative to mineral-based fertilizers. Dolomite is a carbonate mineral is rich in elements CaO and MgO. Dolomite is a double bond between the carbonate of calcium and magnesium, wherein the double compound is calcite (CaCO3) and magnesite (MgCO3) or MgCa (CO3). Dolomite is widely used in agriculture, building materials or in the industry. In nature, these minerals generally always occur together with limestone, quartz, pyrite and clay. Dolomite can be used to neutralize acid soil and to withstand the acidity caused by the use of urea. Dolomite could neutralize the acidity of the soil through the ion, calcium and magnesium cations remove hydrogen ions in the soil. By giving dolomite, soil pH will be increased. A zeolite mineral is a crystalline substance with a structure characterized by a framework of linked tetrahedra, each consisting of four O atoms surrounding a cation. This framework contains open cavities in the form of channels and cages. These are usually occupied by H2O molecules and extra framework cations that are commonly exchangeable. The channels are large enough to allow the passage

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of guest species. In the hydrated phases, dehydration occurs at temperatures mostly below about 400°C and is largely reversible. (Douglas, et al,1997). Based on their high ion-exchange capacity and water retentivity, natural zeolites have been used extensively in Japan as amendments for sandy soils, and small tonnages have been exported to Taiwan for this purpose (Minato,1968). Indonesia has a lot of reserves of zeolite and dolomite. The existence of zeolite and dolomite are scattered in various regions in Indonesia. This condition is an opportunity for small and medium industries (SMEs) to undertake the processing of these minerals as raw materials of mineral-based fertilizers. SMEs can perform the processing of these minerals in the preparation stage of the process of crushing the materials. A crusher is a device that is designed to reduce large solid chunks of raw material into smaller chunks. Crushers are commonly classified by the degree to which they‟re fragment, the starting material with primary crushers that do not have much fineness, intermediate crushers having more significant fineness and grinders reducing it to a fine power. A crusher can be considered as primary, secondary or fine crusher depending on the size reduction factor (Gupta, 2011). During the past decades, impact crushers have become widely used machine for comminution operations because of their high size-reduction ratio, easy modification of the product size distribution and good “cubic” shape of the product (Nikolov,2004). Hammer crusher is one kind of impact crusher, operating principle of the hammer crusher is the hammers are pivoted so that they can move out of the path of oversize material, or tramp metal, entering the crushing chamber. Pivoted hammers exert less force than they would if rigidly attached, so they tend to be used on smaller impact crushers or for crushing soft material. The exit from the mill is perforated, sa that material which is not broken to the required size is retained and swept up again by the rotor for further impacting (Wills,2006). During this time, SMEs minerals have limitations in the choice of means of existing production processes. Crushers such as jaw crusher, cone crusher, hammer crusher and others still obtained by imports. This condition causes a huge investment costs. In addition, the limited spare parts becomes problem in the production process. Provision requires a long time due to indent the process, so that the production process must be stopped when damaged. Sometimes, equipments available are not in accordance with the characteristics of local raw materials. The problem that often occurs in the crushing is the clogging of the sieves.The production process will be influenced by the selection of the right equipments, the purpose of this research is to

design and preliminary tests on hammer crusher can be used in SMEs in accordance with the characteristics of local raw materials. Through this research are expected availability of hammer crusher are made using local components, and economically can be obtained by SMEs. 2. Methods 2.1 Design of hammer crusher The process begins with sketching prototypes, followed by calculation of design parameter, then manufactured of main components and supporting components of jaw crusher and hammer mill. A bending and shearing machine (HACO) used for metalworking. Welding process performed by SMAW method using a welding machine (Black RHINO, MMA 500), manufactured of hammer crusher hammers carried by the metal casting process using induction furnace (INDUCTOTHERM). All stages of the process is done in workshop and laboratories of technical implementation unit of mineral processing, Indonesian Institute of Sciences. Calculation of the theoretical capacity is done by using the equation (Warji,et al,2010) Qt= (1) Hammer mill shaft speed calculation is done by using the equation (Gupta,1980) n2= (2) Determination of length of the belt is given by: (Gupta,1980) L = 2C + 1.57(D2+D1)+ (3) The belt contact angle is given by equation: (Gupta,1980) Sin-1β = (4) The angles of wrap for pulleys are given by: (Holowenko,et al,2004) α1,2 = 180 ± Sin-1 (5) The belt tension can be use equation: (Holowenko,et al,2004) T2 = (6) and T1 = SA (7) Power, torque and shaft diameter calculated based on the equation (Warji,et al,2010) P=

; Tmax =

;d=

(8)

The maincomponent of hammercrusher are the hammers, therefore it needs attention in the design. The hammers are subjected to shear force at the point of fixation, centrifugal force due to rotation, bending force due to striking of the material. When a sudden impact is observed by the blow bars due to input feed striking over, it experiences an impact

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load. The effect of impact loads differs appreciably from that of the static loads as with a suddenly applied load, both the magnitude of the stresses produced and resistance properties of materials are affected, hammer can be made using different section like I section, T section, S section, cylindrical bars, rectangular bars etc. The shape of the hammers decides the impacting capacity as well as the strenght of the crusher (Gupta,2011). 2.2 Preliminary test methods Preliminary test conducted to determine the performance of the hammer crusher. Particle analysis carried out on the resulting product. The particle size analysis is the method used to determine the particle size distribution or the grain size distribution of rock/ore materials. In practice, close size control of feed to mineral processing equipment is required in order to reduce the size effect and make the relative motion of the particles separation dependent (Adepoju et al, 2001). The screening of the crushed rock samples was carried out in a set of sieve using the Laboratory Sieve Shaker. The sieve was arranged in the order of decreasing aperture: 1700, 850, 425, 250, and 180 µm by placing the sieve that has the largest opening at the top and the least opening at the bottom. A tight fitting pan or receiver was placed below the bottom sieve to receive the finest grained which is referred to as undersize. The crushed sample was placed on the top sieve and a lid was used to cover it to prevent escape of the rock sample during the process. The set of the sieve was then placed in a sieve shaker which vibrates the sieve for proper screening. This operation was carried out on each of the rock sample for five minutes. This was achieved by using the automatic control timer of the sieve shaker. After the screening analysis, the retained sample on each sieve was measured on weigh balance and recorded to the corresponding sieve opening size (Olaleye, 2010). 3. Results and Discussion

ofdolomiteand zeolite. From the experiments, obtained a real hammer crusher capacity is 825-900 kg / hour of raw materials (dolomite and zeolite) at 2,900 rpm, and 685-750 kg / hour of raw materials (dolomite and zeolite) at 1,450 rpm. The efficiency gained from the comparison of the number of products produced with the amount of product lost. Of the 21 times repetition shows that the efficiency of hammer crusher is beginning to look steady on repetition of 10 and so on, as it is presented in Figure 3. After in stable condition, at 1,450 and 2,900 rpm obtained the highest efficiency of hammer crusher to process dolomite is 82.11% and 88.54%, for zeolite efficiency is 88.79% and 94.81%. Table 1. Specifications of Hammer Mill Parameter Length of crusher Wide of crusher Height of crusher Length of belt Belt contact angle Tension in the slack side of belt Tension in the tight side of belt Diameter of main shaft Power (electromotor)

Value 1200 600 600 1700 2.91 520.36 1154.41 127 15

Unit mm mm mm mm radian N N mm HP

Figure 1. Double-shaft hammer crusher

Hammer crusher used is a double-shaft hammer crusher. Specification of hammer crusher is presented in tables 1. The design of hammer crusher is presented in Figures 1.Crusher capacity is 1 ton per hour of raw material. The maximum size of the feed material is 80 mm. The hammers and liners are made using manganese steel, composition of the hammers are presented in Table 2. There are 12 pieces of hammers. Each hammer has 2 kilogram of weight, the dimensions of the hammers are presented in Figure 2. Framework of crusher made using U-channel steel (UNP 150), and 10 mm then 3 mm Mild Steel used for crusher chamber and hopper. The pulleys made of gray cast iron. Preliminary testshowed, that the double-shaft hammer crushercanbe usedforthe processing

Figure 2. The Hammers

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Table 2. Composition of The Hammers Elements C Si Mn Elements P S Cr Mo Ni

% Weight 1.78 0.134 12.97 % Weight 0.0966 <0.0002 2.91 0.0065 0.0624

Elements Al Co Cu Elements Fe Zn Se Sn Pb

% Weight 0.00097 0.0262 0.0857 % Weight 81.6 0.0049 0.01240 0.00097 0.0156

Table 3. Particle size distribution of (1450 rpm) Sieve size range (µm)

Sieve fraction Wt (%)

+1700 -1700+850 -850+425 -425+250 -250+180 -180+150 -150

2.07 12.66 10.35 15.87 12.41 35.17 11.46

Dolomite

Nonimal Cumulative aperture size (µm) Undersize Oversize (%) (%) 1700 97.93 2.07 850 85.27 14.73 425 74.92 25.08 250 59.05 40.95 180 46.64 53.36 150 11.46 88.54 0.00 100.00

Table 4. Particle size distribution of Dolomite (2900 rpm) Sieve size range

Sieve fraction

(µm)

Wt (%)

+1700 -1700+850 -850+425 -425+250 -250+180 -180+150 -150

0.21 10.73 8.63 11.57 10.73 45.24 12.88

Nonimal aperture size (µm) 1700 850 425 250 180 150

Cumulative

Undersize (%) 99.79 89.06 80.43 68.86 58.12 12.88 0.00

Oversize (%) 0.21 10.94 19.57 31.14 41.88 87.12 100.00

Table 5. Particle size distribution of Zeolite (1450 rpm) Figure 3. Efficiency of product at 21 repetition charts at various speed rotation From Table 3-6 of particle size distribution, it appears that the number of particles dolomite most established on the size of -180 + 150 μm, amounted to 45.24% by weight at 2900 rpm. For zeolite with the same rotation (2900 rpm) obtained finer size than dolomite, indicated from more particles that pass on the size of -150 µm.Despite of dolomite and zeolite have same hardness value where is 3.5 mosh, the result in mineral processing can be different. From this experiment, chance to achieve particle size of -150 µm for zeolite is much easier than dolomite, with proper and improving of the machine itself. According table 2 until 5, at least 60% of particle distribution in the size of -250+180 µm and above. And the rest still have to be crushed again if it wants to be use for agriculture need. Present day, specification of particle size of mineral-based fertilizer processing is -250+180 µm above. The different rotation speed that is show in this experiment indicated that with higher rotation speed, particle where in size of -425+250 µm got more chance to be crush to become finess. It has shown in Table 4 and 6, with the decreasing of weight percentage in the size range of -425+250 µm.

Sieve size range

Sieve fraction

(µm)

Wt (%)

+1700 -1700+850 -850+425 -425+250 -250+180 -180+150 -150

4.61 17.52 14.27 4.23 15.84 13.85 29.67

Nonimal aperture size (µm) 1700 850 425 250 180 150

Cumulative

Undersize (%) 95.39 77.87 63.59 59.36 43.52 29.67 0.00

Oversize (%) 4.61 22.13 36.41 40.64 56.48 70.33 100.00

Table 6. Particle size distribution of Zeolite (2900 rpm) Sieve size range

Sieve fraction

(µm)

Wt (%)

Nonimal aperture size (µm)

+1700 1700+850 -850+425 -425+250 -250+180 -180+150 -150

2.07 13.93

1700 850

12.65 4.13 12.20 16.78 38.24

425 250 180 150

Cumulative

Undersize (%) 97.93 84.00

Oversize (%) 2.07 16.00

71.35 67.22 55.02 38.24 0.00

28.65 32.78 44.98 61.76 100.00

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In Figure 4 and 5, be repeated 21 times to obtain a preliminary test on the particle size distribution of 180 + 150 μm and -150 lm. Based on the test can be seen that at 2900 rpm have more finer size than at 1450 rpm for both zeolite and dolomite. Zeolite is more easily destroyed than the dolomite to achieve finer particle size. The terrific speed of the hammer produces kinetic energy that is dissipated in the material, causing it to disintegrate. According Rahmadian (2012), the increase rotation of machine affect the result of milling, the faster rotation will increased the results. Higher efficiency can be found at high rotation because material will get high chance to be pounded many times before left the crusher zone. Also double rotation design give more chance to crush material because this design give more dynamic pounded than single rotation, material that enter the range of collision the hammer will get the chance to be thrown and get hit by another hammer many times before it out from crusher zone. The increasing of speed can prolonged the collision of hammer with material before it out the crusher zone.

4. Conclusion Double shaft hammer crusher can be used to crush dolomite and zeolite as raw material for the manufacture of mineral-based fertilizers. The theoretical capacity is 1,000 kg/hour of raw material (dolomite and zeolite). The hammer crusher real capacity was 825-900 kg / hour of raw materials at 2,900 rpm, and 685-750 kg / hour of raw at 1,450 rpm. At 1,450 and 2,900 rpm, obtained the efficiency of hammer crusher to process dolomite was 82.11% and 88.54%, for zeolite efficiency was 88.79% and 94.81%. Acknowledgements Authors acknowledge financial support and facilities from tehnical implementation unit of mineral processing, Indonesian Institue of Sciences. References [1] Adang S.,2011.Mineral- based compound fertilizer as alternative.http://esdm.go.id/berita/56artikel/4133-pupuk-majemuk-berbasis-mineralsebagai-alternatif.html., accessed on 5 June 2015. [2] Adepoju S O.,Olaleye B M., 2001. Assessment of gravity concentration of silica sand from Itakpe iron ore tailings by tabling.Nigerian Journal of Engineering Management 2(2),5155.

Figure 4. Particle size distribution of dolomite chart

[3] Douglas S C., Alberti A.,Armbruster T.,Altiori G.,Colella C., PassagliaE.,et al.1997. Recommended nomenclature for zeolite mineral: Report of the subcommittee on zeolites of the international mineralogical association, commision on new minerals and mineral names. The Canadian Mineralogist Vol 35,1571-1606. [4] Gupta D.,2011.Design and analysis of a horizontal shaft impact crusher. National institute of technology, chapter ,p2. [5] Hall A.,Holowenko A.,Laughlin H.,2004.Theory and Problem of Machine Design . tata Mc Gaw- Hill Publishing Company ltd. New Delhi. 4th Edition Pp6-89. [6] Khurmi RS.,Gupta JK.,1980.A textbook on Machine Design.chapter 17,p647 [7] Minato H., 1968. Characteristics and usesof natural zeolites. Koatsugasu 5,536-547. [8] Nikolov S.,2004. Modelling and simulation ofparticle breakage in impact crushers.International jounal of mineral processing 74S,S219-S225.

Fig 5. Particle size distribution of zeolite chart 1st International Conference on Appropriate Technology Development 2015, Bandung, October 5th-7th, 2015 | 31

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[9] Rahmadian O.,2012. Test performance of hammer mill with corncob feed. Journal of Lampung agricultural engineering .1(1)p:11-16. [10] Olaleye B M., 2010.Influence of some rock strength properties on jaw crusher performance in granite quarry.Mining Science and Technology Vol 20 No.2,204-208. [11] Arji.,Kuncoro S.,Asmara S.,Rahmawati H.,2010.Design and manufacturing hammer mill of cassava.Agriculture Engineering Journal.Vol 8 No2,p:59-66. [12] Will A B.,Munn TN.,2006. Mineral processing technology.An introduction to the practical aspects of ore treatment and mineral. Chapter 6,p136.

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An Appropriate Technology of Composting for Supporting Sustainable Agriculture Adi Mulyanto Institute for Environmental Technology Agency for the Assessment and Application of Technology Telphone: +62-021-75791377; fax: +62-021-75791377. E-mail address:[email protected] Abstract : The conditions of agricultural land in Indonesia are currently classified unhealthy with organic matter content of less than 2%. Healthy soil contains organic materials more than 5%. Soil as a medium for the production of plants must have a healthy condition. To achieve a healthy soil condition it must be done the principle of nutrients recycling. Ideally all the organic material must be returned to the soil, so the soil is able to provide food for the plant. As a result, soil fertility will occur. To accelerate the availability of nutrients in the soil, the organic material should be mineralized through composting process. The uses of compost have several advantages, among others, are able to fertilize the soil and at the same time conserve and nourish the soil ecosystem and avoid the possibility of environmental pollution. To facilit ate and ease of composting process, then made equipment that support the production of compost. This paper will convey the equipment that has been produced to support the production of compost made from organic municipal solid wastes such as shredder, turning machine, screener and mixer. The resulting compost is used as a soil conditioner on agricultural land to grow vegetables. Keywords: healthy soil; composting; shredder; turning machine; screener; mixer 1.

Introduction

Composting process is already known since ancient times and has been done by farmers traditionally. At the time of the introduction of inorganic fertilizers, the habit of making compost becomes unattractive. But at this moment after emerging environmental issues and food security, composting for use as a soil conditioner began to be considered. The use of compost as part of environmentally friendly agricultural activities needs to be promoted or be reminded again in line increasing number of negative environmental impacts that occur as a result of agricultural activities that rely on agricultural chemicals (Sutanto, 2002). Two functions are performed by composting, they are turning nuisancecausing waste product into safe handled material and resulting the valuable commodity, therefore composting is an environmentally beneficial process (Rynk, et al. 1992). Compost is the result of decomposition of organic material through biological processes with the help of decomposing organisms. Decomposition process can take place in aerobic (with air support) or anaerobic (without air support) (Epstein, 1997). The process of composting produces heat. The heat can remove moisture and kill pathogens as well as weed seeds (Rynk, et al. 1992). The main function of the compost is to help improve the soil properties in term of physical, chemical and biological. Physically compost can loosen soil; application compost in soil will increase the number of cavities so that soils become loose. While chemical properties capable addressed by compost application are to increase the cation exchange capacity (CEC) on soil and can

increase the ability of soil to store water (water holding capacity). As for the improvement of the biological properties, compost can increase the population of microorganisms in the soil (Simamora and Salundik, 2006). The superiority of compost contents complete macro and micro nutrient. Macro nutrients contained in the compost among others, N, P, K, Ca, Mg, and S, while the micro-element content among others, Fe, Mn, Zn, Cl, Cu, Mo, Na and B (Stoffella and Kahn, 2001). Rynk, et al. (1992) stated that compost is an ecologically sound product. Manufacture of equipment composting in private companies have the background of the need of compost as soil conditioner for agricultural land owned by the company. At the beginning the company bought compost from other parties. Because the needs of the compost are gradually increasing, the company thought to produce it themselves. With the Western Java Environmental Management Project of the World Bank, the composting capacity should be increased, that is up to 5 tons per day. Therefore, it would require the equipment to produce it. The success of the manufacture of equipment for composting such as shredder, turning machine, screener and a mixer, the company had the idea to produce the equipment and sold. Interested buyers come from the local governments and other private companies. Local Governments who buy domestic waste management equipment for compost production generally have not proper decisions to the placement of the equipment. They placed the equipment in the landfill area. Local Governments build facilities for waste

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treatment activities at considerable cost, but in general the facility is not active or difficult to run. Compost production has a long chain of the management of municipal solid waste. Ranging from problem solving in aesthetic and environmental hygiene; global warming; residues minimization of waste dumped in landfills and at the same time to nourish and conserve the soil ecosystem by applying compost to agricultural land (Sutanto, 2002). Household compost production scale is difficult to do, as well as the production of compost in the landfill area. So that the most suitable of producing compost derived from organic municipal solid waste in Indonesia is done communally. However, composting of solid organic waste from the market should be done with a larger scale since the amount is much higher. 2.

Materials and Methods

Composting is done aerobically with open windrow system. All organic material eventually decomposes both natural and by human intervention. Natural decomposition process can lengthen over period of months or even years. Factors that affect the composting process (Ministry of Agriculture and Food, 1998) were the basic considerations for the design of equipment. The factors are carbon to nitrogen ratio (C/N ratio); surface area and particle size; aeration; porosity; moisture content; temperature; pH of materials; nutrients and toxic substances. Surface area and particle size influences the composting process for microbial activity happens at the interface of raw material surfaces and air. Particle size largely determines the amount of space between the materials (porosity). Enough pores will allow air and water dispersed more evenly in the stack. To increase the surface area can performed by reducing the particle size of the material, which optimal particle size of 2-10 cm. Large particle size would hinder aeration and microbial performance so the maturation process will take longer. The increasing contact between microbes and the material causing decomposition process will also be faster (Jeris and Regan, 1993). To achieve the smaller particle size, the material necessary to be shredded. Therefore, it is necessary manufacture thrasher or shredder. Factors C/N ratio, aeration, porosity, moisture content, and temperature can be set at the optimum condition if done the turning of the raw material compost pile. On a small scale, compost heaps able to turned manually. But the large scale required for turning machine. Turning activity can be used as a means of mixing. C/N ratio should be evenly distributed throughout the compost pile material. Rapid decomposition can occur under conditions of sufficient oxygen. Aeration will naturally occur

when the temperature increases. Warm air will come out and cooler air will enter into the compost pile. Aeration is determined by the porosity, particle size and moisture content of the material (humidity). If aeration is blocked, anaerobic process will occurred and will produce foul-smelling ammonia. Aeration can be enhanced by turning or injecting air into the compost heap (Jeris and Regan, 1993). Porosity is the spaces between the particles of compost that contains water or air. Air will supply oxygen to the composting process. If the spaces filled with a lot of water, then the oxygen supply is reduced and the composting process will be disrupted. The porosity of compost pile will reduce because of compacting (Ministry of Agriculture and Food, 1998). Turning the heap can increase the porosity. Humidity plays an important role in the metabolism of microbes and indirectly affects the supply of oxygen. Decomposing organisms can utilize organic material when organic materials are soluble in water. Humidity 40-60% is the optimum range for microbial metabolism. If the humidity below 40%, microbial activity will decline. If the humidity is greater than 60%, then the nutrients will be leached and the air volume is reduced, resulting in microbial activity will decrease and there will be anaerobic fermentation. Therefore, keeping the water content so that the humidity is ideal for composting is very important. (Jeris and Regan, 1993). The heat generated from microbial activity. An increase in the temperature has a straight relationship with the oxygen consumption. The higher the temperature, the more the consumption of oxygen and the sooner will the process of decomposition. Temperatures between 30º-60ºC showed rapid decomposition activity. While temperatures higher than 60 °C will kill most microbes and only thermophilic microbial which can survive. High temperatures will also kill the plant pathogenic microbes and seeds of weeds. When the temperature has reached 70ºC, then immediately turns the pile to reduce the temperature (ibid). The composting process can occur in the pH range 5.5 - 9. The composting process will cause changes in the pH of the composted material. For example, the process of acid leaching temporarily or locally will cause a decrease in pH (acidification), while the production of ammonia from compounds containing nitrogen will increase the pH in the early phases of composting. pH levels of mature compost is usually near neutral (Epstein, 1997). Contain enough nutrients in the form of phosphorus and potassium is essential to be added to a compost pile. In general, materials that contain both elements are cow manure and chicken droppings (Ministry of Agriculture and Food, 1998). Some organic materials may contain substances that are harmful to microbial life. Heavy metals such as Mg, Cu, Zn, Ni, Cr and Pb are some materials that belong to this category. Heavy metals

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are not biodegradable and will remain there. The heavy metals can come from contaminated organic material or other garbage. Water also can be a medium to contaminate the compost materials with heavy metals. Hazardous pollutants can come from waste batteries, paint, and others. These heavy metals can affect the work of microbes in breaking down organic matter (Paulin and O'malley, 2008).

be generated. The size reduction can be achieved because of pounding action. At the end of the blades row mounted a type of fan to accelerate the release of garbage that has been shredded. Several types of shredders and their specification can be seen in table 1.

The data used for the design of the equipment taken from the literatures (Green, 2008; Reynolds, 1982; Foust, et al 1980). 3.

Results and Discussion

3.1 Shredder Shredder serves as a tool to reduce the particle size of the material to be composted (Figure 1). This tool can also be used to crush mature compost produced. This device consists of hopper to put organic waste and the blade that also functions as a screw to bring the shredded garbage out of the equipment. The blades welded to the axle that rotates at a speed of 1500 rpm. Installations of the blades have an angle of 10 degrees so that the function of the screw can

Figure 1. Shredder

Table 1. Specification of shredders.

3.2 Turning machine For small scale (hamlet/RT and neighbourhood/RW) turning of the compost pile is done manually, while for large scale (village and market) need to use turning machine to ease the job. Small-scale compost heaps is usually equipped with air tunnel made from wood or bamboo to enhance aeration system (Figure 2). Illustration of aeration enhancement mechanisms can be described as follows: The inside of the compost pile has a hot

temperature. This causes the air trapped inside will flow naturally 4 upward, as a result the air pressure in the stack is reduced so that fresh air will enter into the compost pile. This process can be described as the chimney effect. To form a neat compost pile, need to be made rectangular mould made of wood. Compost pile is prepared from below, and then step by step the mould lifted upwards.

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Figure 2. Tunnel For large scale, the fulfillment of the adequacy of oxygen should be carried out through the process of turning compost pile using the machine. First generation turning machine was made by utilizing the junk (Figure 3). Driving machine to drive the turning machine utilizes diesel engine. For steering, clutch and gearshift utilized the used tractors. To turn the compost used electromotor power which has rotation of 1440 rotations per minute (rpm) and connected with gear box to reduce the rotation of up to about 360 rpm. Electromotor has a power of 10 hp. This turning machine has a weakness in terms of capacity because it is only capable of turning the compost pile as high as the maximum of 60 cm. With the demands of compost production capacity of 5 tons per day, then the turning system needs to be replaced with a larger capacity. To increase the capacity of turning, then we made the second generation turning machine (Figure 4).

Figure 4. Turning machine (recondition forklift) Second generation turning machine was constructed based on reconditioned forklift. At the fork mounted bucket equipped with a hydraulic system. The bucket rests on a hinge at one side. Operation of this machine is the same as the operation of the forklift. The bucket lifts the compost materials from the windrow and spills them down again. This process cause mixing the materials and improve the mixture into a loose windrow. By turning the materials, compaction process can be avoided and oxygen requirement can be fulfilled. The most important part to be maintained is the machine itself and the hydraulic systems. Advantages of this second generation turning machine are able to produce compost with a greater capacity, more resilient, and easier operation than the first-generation one. The turning machine can also be used for transporting, stockpiling and loading raw materials and finished compost products with the maximum purpose of facilitating the movement of materials through the compost facility. The size of the compost material that cannot be reduced is a disadvantage of this second generation turning machine.

Figure 3. Turning machine Table 2. Specification of turning machine.

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3.3 Screener Screener has a conical shape with different diameters at both ends and classified as rotating or trommel screener. The basic design of the trommel screen consists of diameter, length and inclination of cones. The diameter specifies the amount of surface area to which the compost is contacted and the length and inclination of the screen specifies the retention time within the screen. The cones has 2 sizes of diameter, one end has a diameter of 165 cm and the other end 50 cm (Figure 5). The length of the cones is 300 cm. The compost will be screened entered from the small diameter section. Different diameter makes inclination, so that the material will be able to flow as it rotates. Rotary screen rotates on its axis at a speed of about 17 rpm. With such speeds, the material does not come round to the top of the filter, so that the effect of the wind to blow away the compost can be minimized. Driving machine used is a diesel engine. But electromotor can also be used depending on the choice. Between the engine and gear box connected with a belt and pulleys, while the gear box and axle of screener connected by a chain and sprockets. The screener is not made cylindrical, but made eight squares to create a collision for effective screening. This tool is made with the intention to produce compost which is relatively uniform size and suitable for use in largescale composting. When the mature compost has a size greater than 10 mm, the compost needs to be reduced in size using a shredder or be returned again as a bulking agent in the compost pile. Shredded compost is then filtered. On a small scale, this equipment should not necessary be there. To homogenize the compost produced only require a simple screening tool, in the form of woven wire sieve with holes of a maximum of 10 mm equipped with a frame that is placed at the edges.

Table 3. Specification of compost screener.

3.4 Compost Mixer For applications to agricultural land, sometimes compost needs to be mixed with inorganic fertilizer to make balanced fertilizer. The type of this equipment is double ribbon mixer (Figure 6). Compost and fertilizer to be mixed is inserted through the door on top of the mixer. The resulting mixture was removed from the bottom by opening the lid that is located at the bottom of the mixer. This equipment works in a batch system where the measured volume or weight of materials such as compost and inorganic fertilizers enter into the mixer in accordance with a prescribed mixed ratio. The materials are then blended for a period of about 5 minutes. Materials in the mixer move from left and right sides toward the middle and vice versa to create turbulent conditions.

Figure 6. Compost mixer

Table 4. Specification of compost mixer

Conclusions

Figure 5. Compost screener.

Agricultural land in Indonesia has been classified as a sick soil. The soil remediation must be done immediately, one of them by way of adding organic matter in the form of compost derived from municipal solid waste. This step can solve problems that involve many aspects, among others, are healthy agricultural lands, reducing the amount of greenhouse gases in the form of methane gas, and extend the lifespan of landfills. Government needs to

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continuously provide the initial facilities and guidance to the communal municipal solid waste management in Indonesia. Equipment that has been donated is generally placed in a location that is not proper, namely in the landfill. As a result the equipment is not used because it was unable to process all existing waste. Selection of the appropriate composting equipment based on its capacity is very important. Acknowledgements The author would like to thank CV. Agro Duta, Bandung which has facilitated and accommodate all thought that the expectations of “an idea into a transaction" can be realized. Reference [1] Epstein, E., 1997. The Science of Composting. Technomic Publishing Inc. Pensylvania. [2] Foust, A.S., Wenzel, L.A., Clump, C.W., Maus, L., Andersen, L.B., 1980. Principles of unit operations. John Wiley & Sons, New York. [3] Green, D.W. ed., 2008. Perry‟s chemical engineers‟ handbook. 8th edition. McGrawHill, New York. [4] Jeris, J. S. and R. W. Regan., 1993. Controlling Environmental Parameter for Optimum Composting. Compost Science 14(1):10-15. [5] Ministry of Agriculture and Food, 1998. B.C. Agricultural Composting Handbook. Composting Factsheet, British Columbia. [6] Paulin. B. and O'malley, P., 2008. Compost Production and Use in Horticulture. Department of Agriculture and Food. Government of Western Australia. [7] Reynolds, T.D., 1982. Unit operations and processes in environmental engineering. Brooks/Cole Engineering Division, Monterey, California. [8] Rynk, et al., (1992). On-farm composting handbook. Northeast Regional Agricultural Engineering Service, Ithaca, NY. [9] Simamora, S. dan Salundik. 2006. Meningkatkan Kualitas Kompos. Agromedia. Jakarta. [10] Stoffella, P. J. and Kahn, B.A., 2001. Compost Utilization in Horticultural Croping System. Lewis Publishers. Washington D. C. [11] Sutanto, R., 2002. Penerapan pertanian organic: Pemasyarakatan dan pengembangannya. Penerbit Kanisius, Yogyakarta.

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Runoff Harvesting as One of Appropriate Technology in Integrated Dry Land Farming Nurpilihan Bafdal1, Sophia Dwiratna NP2 1)

2)

Professor on Soil and Water Engineering Laboratory of Padjadjaran University Ph.D Student on soil and water engineering concentration of Padjadjaran University Jln. Raya Bandung – Sumedang Km. 21 Jatinangor 40563 Email : [email protected]; [email protected]

Abstract: Increased productivity of dry land farming in Indonesia is still limited by the lack of water supply, especially during the dry season; while most regions in Indonesia have high rainfall (over 1500 mm per year). The fact indicates most of the rain that falls on the land surface lost as runoff. The runoff management in dry land farming through runoff harvesting makes it possible to exploit this potential to meet the crop water requirements in the dry season. This research aims to increase productivity and carrying capacity of dry land through appropriate runoff harvesting technology as a source of irrigation water in the area of dry land agriculture. The method used in this research is descriptive analysis method and field observations. The result shows that the slope of the land in the catchment area with coconut + seasonal crop patterns positively correlated to runoff occurs, but not so in the land with a single or mix of seasonal crop which is more affected by the condition of the plant cover. The application of runoff harvesting technology on dry land farming be able to irrigate land planted with maize cultivation area at least 44% of the total catchment area. Runoff harvesting can use as one of alternative appropriate technology for solve the problem lack of water in dry land farming. Keywords: Apropriate technology, Runoff Harvesting, Integrated Dry Land Farming, Irrigation 1. Introduction The high annual rainfall in Indonesia is more than 1500 mm per year; it impacts on the amount of abundant runoff, especially on sloping land. Without special treatment such as runoff harvesting technology, the runoff becomes uncontrollable and can not be used to meet the crop water requirements, especially in the dry season. Runoff harvesting technology in dry land is the technology of collecting runoff from the land surface to be used as an alternative source of irrigation water, espescially during dry season (Prinz and Malik, 2002) Dry land farming has a high potential if it managed properly; one of the lack of dry land farming is the water requirements of plants rely on water from rainfall (rainfed agriculture). This situation is more noticeably or more significant during the dry season; to overcome this, the management of irrigation water is needed so that plants can grow optimally. The characteristics of dry land farming are: (a) low productivity because it relies on rainfall; (b) the plants grown are corn, upland rice, cassava, sweet potatoes, beans with corn as a major crop; (c) mixed cropping as a strategy in anticipation of crop failure; (d) technology is based on a traditional low input farming; (e) the limitation of land use due to the labor constraints (Bafdal, 2013). Reviewing of dry land farmers in Jatinangor that depending their life on dry land farming but it has low productivity, it is need technological breakthroughs, especially to meet the crop water requirements, especially in the dry season. One of the alternatives that can be studied is the application of appropriate runoff harvesting technology to meet the crop water requirements,

especially in the dry season. This research aims to increase productivity and land use of dry land through appropriate runoff harvesting technology as a source of irrigation water in the area of dry land agriculture. 2. Methods The research was conducted with survey and descriptive analytic method for analyzing the potential precipitation on the location of research and field observation method for calculating the potential runoff. Runoff measurements in the field is done by using the runoff plots on four plots with the different cropping pattern and land slope as can be seen in Table 1. Table 1. Cropping patterns, slope and catchment area of runoff plots were used Crop Pattern Coconut + seasonal crop (CSC) Coconut + seasonal crop (CSC) Single Seasonal crop (SSC) Mix Seasonal crop (MSC)

Slope (%) 21 % 23 % 18 % 22 %

Catchment area (m2) 88 66 66 66

The parameters observed in the field are volume of runoff on plots and high rainfall that occurs every runoff measurement. Runoff coefficient values calculated by the equation: C = RO/P (Crithley, 1991) ............................... (1) where C is the runoff coefficient values; RO is the runoff (mm) and P is the depth of rainfall that occurred (mm). To calculate the potential for runoff that can be harvested in the catchment area used the equation:

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V = R x C x A (Lancaster, 2006) ..................... (2) Where: V = the volume of runoff that could be harvested (m3); P is the annual rainfall (m) and A is the catchment area (m2). 3. Results 3.1 Analysis of Regional Rainfall An understanding of the pattern and distribution of rainfall in the location of research became the basis for determining the required supplementary irrigation period is also the basis in determining the potential of runoff that can be harvested in an agricultural area. Analysis of the rainfall region calculated by the method of Thiessen. Figure 1 below shows the rainfall in the research location 97% is determined by rainfall data from Pedca station and 3 percent was determined by rainfall data from Jatiroke Station.

Figure 2. Distribution of Monthly Rainfall in Jatinangor 3.2 Rainfall - Runoff Analysis of the Several Crop Patterns and Slope Actual rainfall and runoff in the field measured during the growing season. Results of field observations indicate that the surface runoff is strongly influenced by rainfall. Figure 3 below the surface runoff that occurs in all the plots were tested increased along with the high rainfall that occurred. Besides influenced by rainfall, the amount of runoff that occurs on farms is also affected by the slope and cropping patterns that are selected. Table 2 below shows the surface runoff in some the cropping pattern and land slope as a result of field observations. Table 2. Surface Runoff in Several the cropping patterns and slope Slope (%) Coconut + seasonal crop (CSC) 21 % Coconut + seasonal crop (CSC) 23 % Single Seasonal crop (SSC) 18 % Mix Seasonal crop (MSC) 22 % Crop Pattern

Figure 1. Thiessen polygon map of the study site Based on the analysis of rainfall, Jatinangor research center has annual rainfall of 1879.69 mm. Based on the amount of annual rainfall, then the location of the study are included in the category of dry land with wet climates where rainfall> 1,500 mm / year (Dwiratna, 2010). Meanwhile, based Oldeman climate classification known that Jatinangor region has five of wet months (NDJFM) where monthly rainfall> 200mm; four dry month (JJAS) where monthly rainfall <100 mm and three humid months (monthly rainfall between 100 -200 mm per month), as can be seen in Figure 2. The condition of rainfall distribution in Jatinangor caused farmers can only cultivate once a year with intercropping pattern between sweet potato + corn + cassava.

Runoff Runoff (mm) coefficient 195.89 0.129 223.26 0.120 251.18 0.152 138.26 0.093

Based on Table 2 is known that in dry land farming with cropping patterns coconut + seasonal crop (CSC), surface runoff is affected by the land slope. The greater slope getting greater the runoff that occurs. But it is different with the runoff on single seasonal crop patterns (SSC) and the mix seasonal crop (MSC). Runoff that occurs on single seasonal crop is greater than the mix seasonal crop despite of having values lower slope. This proves that the runoff is also determined by the crop canopy, which is in the dense crop canopy (MSC) less runoff that occurs, and vice versa. Observations of the rainfall and runoff actual in the field are done to calculate the coefficient of runoff from the catchment area. Runoff coefficient value calculated each time the field observations using equation 1. Table 2 also shows that the smallest runoff coefficient value occurs in mix seasonal cropping pattern, while the largest coefficient value occurs on land with a single seasonal crop pattern.

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Figure 3. The Actual of Rainfall and Runoff Surface in the Field Pontential Runoff that Could be Harvested from the Catchment Area Potential runoff is calculated after the runoff coefficient is known. The amount of water that could be harvested is calculated using the equation 2. The table below shows the results of the calculation of the potential runoff in plots that were attempted. If it is assumed the crop water requirements amount of corn during the growth period is 400 mm (120

days cultived of corn + 4mm/day potential evapotranspiration and corn Kc is 0.83), then at least runoff harvested on a plot with a mix of seasonal crop can be used to irrigate 28.85 m2 of land that planted with maize. The larger the catchment area is used, the greater area that can be irrigated. Therefore, the runoff water harvesting technology can be used as an effective technology in increasing the productivity of dryland farming.

Table 3. Potential Runoff That Can Be Harvested

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4. Conclusions  The slope of the land in the catchment area with coconut + seasonal crop patterns positively correlated to runoff that occurs, but not so in the land with a single or mix of seasonal crop which is more affected by the condition of the plant cover.  The appropriate runoff harvesting technology in dryland farming able to irrigate the land at least 44% of the catchment area.  Runoff harvesting can use as one of alternative the appropriate technology for solve the problem lack of water in dry land farming References [1] Critchley, Will, Siegert, Claus. 1991. Water Harvesting. A Manual for the Design and Construction of Water Harvesting Schemes for Plant Production. Food And Agriculture Organization Of The United Nations - Rome [2] Dwiratna, Sophia. 2010. Monthly Rainfall Stochastic Model and Its Application in Schedule and Planting Pattern Determination of Dryland Farming in Bandung regency. Research Report, University of Padjadjaran. Bandung [3] Bafdal, Nurpilihan; Dwiratna NP, Sophia,; Amaru, Kharistya. 2013. Assess the applicability Run off Management Integrated Farming to Improve Productivity and Capability Dryland. Research Report, University of Padjadjaran. Bandung [4] Lancaster, Brad. 2006. Rainwater Harvesting for Drylands and Beyond, Vol.1.Rainsource Press. Tucson. Arizona [5] Prinz, Dieter and Dr. Amir H. Malik. 2002. Runoff Farming. Institute of Water Resources Management, Hydraulic and Rural Engineering: Germany

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Pilot Scale Technology for Production Organic Biofertilizer Powder Starter to Support Sustainable Agriculture Development Nur Laili, Sarjiya Antonius, Yayuk Kartika, Dwi Agustiyani Research Center for Biology, Indonesian Institute of Sciences (LIPI) Jl. Raya Jakarta-Bogor Km.46, Cibinong 16911, Indonesia Email: [email protected] Abstract: Application of organic biofertilizer is rising due to enhancement in nutrient uptake efficiency and society demands for more green technologies for sustainable agriculture development. The aims of this research were to production of organic biofertilizer powder starter and develop it’s technology that can adopted by industries. We improve a technology to produce biofertilizer powder starter, including bacterial fermentation in pilot-scale fermentor, cells harvesting with continuous centrifuge, freeze-drying, mixing carrier and packaging. We fermented three bacteria which is have the best potency for plant growth promoting, including Bacillus sp. 140B as biocontrol, Brevundimonas sp. AA2 with the highest growth hormone production, and Brevundimonas sp. Piko as phosphate solubilizer. To maintenance and enhance the viability and stability of the cells during freezedrying process, we added combination of skim milk and trehalose as cryoprotectant agents. The cryoprotectants have a good protection for the viability of bacteria, were shown by stability of bacterial population. Carrier materials were used to make powder starter, including maltodextrin and maize starch. Keywords: biofertilizer; carrier material; cryoprotectant agent; powder starter production 1. Introduction Environmental issues in agriculture and agroforestry practices are forcing the farmers to develop methods of cultivation that have a lower impact on the environment. The reduced use of chemical fertilizers with increased application of organic biofertilizers is considered a compulsory route to alleviate the pressure on the environment derived from agriculture practices. The increasing need for environtmentally agriculture practices is driving the use of organic biofertilizer. Biofertilizers are substances which contain beneficial living microorganisms, when applied to seed, plant surfaces, or soil, colonize the rhizosphere or the interior of the plant, enhance the solubility of soil nutrients, promote growth and having potential to increase the yield of crops (Malusa et al., 2012; Kumar, 2014). Biofertilizer is a product which is largely used today in various crops cultivation to replacement of the synthetic fertilizer. The interest in the application of biofertilizer is rising due to enhancement in nutrient uptake efficiency and society demands for more green technologies in sustainable agriculture (Malusa et al., 2007; Adesemoye et al., 2008). Our biofertilizer product consist of potential microorganisms as plant growth promoting rhizobacteria (PGPR), including Bacillus sp., Pseudomonas sp., Brevundimonas sp., Bulkholderia sp., and Brucellaceae sp. Those bacteria have excellent multi activities, include growth hormone production, phosphate solubilization, biocontrol, nitrogen fixation, and also protease activity which is can producing organic acids. Generally Bacillus and Pseudomonas are involved asphosphate solubilizing bacteria plays a major role in the solubilization and uptake of native and applied soil P (Krishnaveni,

2010). Phosphate is essential for early establishment and better growth of plants and responsible for biological rescue capability of solubilizing the insoluble inorganic phosphorus of soil. Nitrogen fixing bacteria also have important role to stimulate the density and length of root hairs, increase the growth through hormonal production, increase biomass, and increase survival rate (Kumar, 2014). The aims of this research were to production of organic biofertilizer powder starter and develop it‟s technology that can adopted by industries. The availability of this starter is expected to increase production of organic biofertilizer. We tried to develop a technology to produce powder starter, include fermentation of bacteria, cell harvesting using continuous centrifuge, freezing process using freeze-dryer, mixing with suitable carrier and packaging. Powder starter have any benefits than liquid starter, include effectivity of handling, packaging and distribution to users, good stability and viability, and long lasting storage. Freeze-drying is a commonly used technique for the production of dried powder starter of organic biofertilizer. Freezing is a complex and glassy system, which is cells are exposed to damage from the process conditions such as very low freezing temperatures and dehydration (Jalali, et al., 2011; Dave et al., 2012). Cells are first frozen to below the critical temperature of the formulation, and then dried by sublimation under high vacuum in two stages, include primary drying, during which unbound water is removed and secondary drying, during which the bound water is removed. To avoid the damage of cell wall or cell death, the presence of cryoprotectants in the drying medium needed. It is important to increase the viability after drying and optimize the production process. After freeze-

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drying, cell will mixed with suitable carrier. Suitable carrier should be cheap, easily used, mixable, packageable, available, and must be non-toxic either to the bacteria or to the plant. Variety of materials used as carriers has been shown to improve the survival and biological effectiveness of inoculants by protecting bacteria from biotic and abiotic stresses (El Fattah et al., 2013).

2.3 Carrier mixing

2. Methodology

2.4 Enumeration of viable cells

2.1 Fermentation and cell harvesting Selected strains, consist of Bacillus sp. 140B, Brevundimonas sp. AA2 and Brevundimonas sp. Piko were subcultured in 50 ml nutrient broth (NB) medium for 24 hours. Then, the culture was inoculated into 1 L NB medium and incubated on rotary shaker for 24 h. After incubation, the culture transferred into 20 L NB in fermentor for 24 to 48 h until stationary phase was reached (Figure 1a). The agitation of fermentor was set at 100 to 150 rpm, the temperature was set at 30 C and the pH controlled at 7.0. Sampling was conducted every 6 hours after inoculation and analyzed for optical density using spectrophotometer at  = 436 nm and bacterial plate counts. When the stationary phase was reached, the culture was harvested using continuous centrifuge at 21000 rpm (Figure 1b). Aftter centrifugation, the pellet cell was collected and transferred into sterile petri dish.

a

Freeze-dried cells were mixed with carrier in sterile condition at three different concentration. We used two types of carrier, include maltodextrin and maize starch. Maltodextrin was sterilized in oven at 80 C for three times, while maize starch was sterilized by autoclaving.

The viability of cells were performed through total plate count technique using nutrient agar (NA) medium. The numbers of viable cell were counted at cell harvesting, before frozen at -80 C, before and after freeze-drying process, freeze-dried cells during storage and after mixed with carrier. Bacteria colonies on NA medium were enumerated and the mean number of bacteria was expresses as CFU/ml and CFU/g. 3. Results and Discussion The fermentation of Bacillus sp.140B and Brevundimonas sp. AA2 was stopped after 48 h, while Brevundimonas sp. Piko was stopped after 30 h from inoculation. Based on growth curve of those bacteria, stationary phase of Bacillus sp. 140B was reached after 42 h, Brevundimonas sp. AA2 after 36 h, and Brevundimonas sp. Piko after 30 h from inoculation (Figure 2). After stationary phase was reached, we harvested the culture from fermentor using continuous centrifuge. Then, the pellet cells were added cryoprotectant agents, consist of skim milk and trehalose. The pellet cell total and it‟s concentrations after centrifugation and added cryoprotectant agents were shown in Table 1.

b

Figure 1. Culture fermentation in fermentor (a); cell harvesting use continuous centrifuge (b) 2.2 Freeze drying Freeze-drying process using method from Savini et al., (2010). The pellet cells were mixed with cryoprotectant agent in a ratio 1:5 (1 ml of cryoprotectant for 5 g of concentrated cells). Cryoprotectant agents were prepared in phosphate buffer saline (PBS) solution at the concentration 20% (w/v) and sterilized with autoclave. We used combination of two cryoprotectant agents, consist of skim milk and trehalose. The suspension was frozen at -80 C for 24 h. Then, sample was dried in a Telstar Lyoquest freeze-dryer with a condenser temperature of -50 C and a chamber pressure P < 1.0 mbar for 24 to 48 h.

a

b

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freeze-drying. While the live cells of Brevundimonas sp. Piko couldn‟t counted before freeze-drying, but the microbial population after freeze-drying shown a high population. After two months storage, the freeze-dried cells of those bacteria shown a good viability by stable microbial population, even Brevundimonas sp. Piko have enhancement 1 log scale.

Figure 2. Growth curve of Bacillus sp. 140B (a); Brevundimonas sp. AA2 (b); and Brevundimonas sp. Piko (c) After frozen at -80 C for 24 h, the cell pellet already for freezing process in freeze-dryer. The viability of the cells were measured using total plate count method before and after freeze-drying process, also during storage for two months later at 4 C. The obtained freeze-dried cells and it‟s concentration were shown in Table 2. Based on data presented in table 1 and 2, it‟s give a variety result. The live cells of Bacillus sp. 140-B declined less than 2 log scale after frozen at -80 C, but the concentration increased 1 log scale after freeze drying. The live cells of Brevundimonas sp. AA2 shown a different result, it‟s increased 1 log scale after frozen and declined 1 log scale after

According to the result of freeze-drying process, it‟s shown that the cryoprotectant consist of skim milk and trehalose gave a good protection for the cells during frozen, freeze-drying process and storage. This is in agreement with findings of previous research by Zayed et al. (2004) reported that combination of skim milk with trehalose greatly enhanced the survival of Lactobacillus salivarius upon freeze-drying and storage. Freeze-drying can destroy membrane structure and change cell viability, but the availability of skim milk and trehalose as cryoprotectant increase the membrane intactness and fluidity (Li et al., 2011). After we obtained the freeze-dried cells, we mixed it with suitable carrier, including maltodextrin and maize starch. We tested three different concentration of bacteria (0.01, 0.05 and 0.1 g bacteria/100 g carrier) and measured the viability of bacteria after carrier mixing and during storage for 1 week at 4 C and 25 C. Total counts of bacteria cell in carrier were shown in Table 3 and 4.

Table 1. The pellet cell total and it‟s concentrations of starter bacteria Bacteria isolates

Volume (L)

Cell pellet total (G)

Bacillus sp. 140-B

19 L

Brevundimonas sp. AA2 Brevundimonas sp. Piko

Concentration (CFU/ml) After Centrifugation

After added cryoprotectants

24.478 G

50x 1012

77 x 1012

21 L

69.972 G

34 x 1011

66 x 1012

17 L

44.571 G

~

~

Table 2. Freeze-dried cell total and it‟s viability of bacteria after freeze-drying and two months storage at 4 C Bacteria isolates

Freeze-dried cell total (G)

Bacillus sp. 140-B

Concentration (CFU/g) freeze-

During storage months (4 C)

Before freezedrying

After drying

4.86

74x 1010

236 x 1011

92 x1011

Brevundimonas sp. AA2

20.82

69 x 1013

86 x 1011

268 x 1011

Brevundimonas sp. Piko

11.55

~

143 x 1011

83 x 1012

for

2

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Table 3. Population and viability of bacteria in maltodextrin carrier Bacteria isolates Bacillus sp. 140-B

Concentration 0.01 0.05 0.1 0.01

Brevundimonas sp. AA2

Brevundimonas sp. Piko

0.05 0.1 0.01 0.05 0.1

Bacterial population (CFU/g) After carrier mixing Storage at 4 C Storage at 25 C Not detected 200 x 106 Not detected Not detected Not detected Not detected 8 Not detected Not detected 35 x 10 8

Not detected

44 x 10 9

45 x 10

236 x 10 9

Not detected

6

Not detected

6

231 x 107 Not detected Not detected Not detected

30 x 10 Not detected Not detected Not detected

31 x 10 Not detected 9

60 x 10 Not detected

Table 4. Population and viability of bacteria in maize starch carrier Bacteria isolates Bacillus sp. 140-B

Brevundimonas sp. AA2

Brevundimonas sp. Piko

0.01 0.05 0.1 0.01

Bacterial population (CFU/g) After carrier mixing 70 x 106 Not detected Not detected 8 44 x 10

0.05

196 x 10

0.1 0.01 0.05 0.1

75 x 10 Not detected Not detected Not detected

Concentration

Results presented in Table 3 and 4 show that carrier mixing process was not optimally. There was any problem for carrier mixing technique or method and caused the live cells of bacteria in carrier declined. There was also shown that total counts of bacterial population not stable. According to this results we need to select better carrier material, effectiveness of carrier mixing technique and try to make combination of carrier materials to optimize and maintenance of bacteria cell‟s viability for long period of storage. 4. Conclusion We improve a technology to produce biofertilizer powder starter, including bacterial fermentation in fermentor controlling temperature, pH and agitation, cells harvesting with continuous centrifuge, freezedrying process, mixing carrier and packaging. Cryoprotectant agents, consist of combination skim milk and trehalose were added in freeze-drying process to give protection to bacteria cells and increased it‟s viability during process. Viability of bacteria was measured using total plate count method at cell harvesting, before frozen at -80 C, before and after freeze-drying process, freeze-dried cells during storage and after mixed with carrier. Combination of skim milk and trehalose as cryoprotectant agent gave a good effect for the cells,

7 7

it was shown by stability of bacterial population during frozen, freeze drying and storage. Carrier materials were tested in this research, including maltodextrin and maize starch, but it‟s didn‟t show a good result, so we must have try to select carrier material and effectiveness of carrier mixing technique. Acknowledgment This research has been performed in the Agriculture Microbiology Laboratory, Microbiology Division, Research Center for Biology, Indonesian Institute of Sciences (LIPI) and financially supported by Commercial Product Program, Flagship Program from Deputy for Natural Sciences, LIPI 2015. References [1]

Adesemoye, A.O., H.A. Torbert & J.W. Kloepper. 2008. Enhanced plant nutrient use efficiency with PGPR and AMF in an integrated nutrient management system. Canadian Journal of Microbiology, 54(10): 876-886.

[2]

Dave, R., R. Patel, J. Patel & H. Chauhan. 2010. Effect of cryoprotectant on lyophilisation of doxorubin-HCl loaded

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chitosan nanoparticles. International Journal of Pharmacy and Life Science, 3(6): 17691772. [3]

El Fattah, D.A.A., W.E. Eweda, M.S. Zayed & M.K. Hassanein. 2013. Effect of carrier materials. Sterilization method, and storage temperature on survival and biological activities of Azotobacter chroococcum inoculant. Annals of Agricultural Science, 58(2): 111-118.

[4]

Jalali, M., D. Abedi, J. Varshosaz, M. Najjarzadeh, M. Mirlohi & N. Tavakoli. 2012. Stability evaluation of freeze-dried Lactobacillus paracasei subsp. Tolerance and Lactobacillus delbrueckii subsp. Bulgaricus in oral capsules. Research in Pharmaceutical Sciences, 7(1): 31-36.

[5]

Krishnaveni, M.S. (2010). Studies on phosphate solubilizing bacteria (PSB) in rhizosphere and non-rhizosphere soils in different varieties of foxtail millet (Setariatalica). International Journal Agricultural and Food Science Technology, 1(1): 23-39.

[6]

Kumar, V. 2014. Characterization, bioformulation development and shelf-life studies of locally isolated biofertilizer strains. Octa Journal of Environmental Research, 2(1): 32-37.

[7]

Li, B., F. Tian, X. Liu, J. Zhao, H. Zhang & W. Chen. 2011. Effect of cryoprotectant on viability of Lcatobacillus reuteri CICC6226. Applied Microbiology & Biotechnology, 92: 609-616.

[8]

Malusa, E., L. Sus-Paszt & E. Zurawicz. 2007. The effect of a mycorrhiza-bacteria substrate and foliar fertilization on growth response and rhizosphere pH of three strawberry cultivars. International Journal of Fruit Science, 6(4): 25-41.

[9]

Malusa, E., L. Sus-Paszt & J. Ciesielska. 2012. Technologies for beneficial microorganisms inocula as biofertilizers. The Scientific World Journal, 1-12.

[10]

Savini, M., C. Cecchini, M.C. Verdenelli, S. Silvi, C. Orpianesi & A. Cresci. 2010. Pilot-

scale production and viability analysis of freeze-dried probiotic bacteria using different protective agents. Nutrients, 2; 330-339. [11]

Zayed, G. & Y.H. Roos. 2004. Influence of trehalose and moisture content on survival of Lactobacillus salivarius subjected to freezedrying and storage. Process Biochemistry, 39: 1081-1086.

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Influence of urea granulated Zeolite and Nitrification Inhibitors on Growth of Maize (Zea mays L. Var. B8) 1)

Oslan Jumadi* , Ratna Dewi 1), Andi Takdir Makkulawu2), R. Neni Iriany2), Yusminah Hala1), Hartono1), St. Fatmah Hiola1), Kazuyuki Inubushi3) 1)

Department of Biology, Faculty of Mathematics and Natural Sciences,State University of Makassar. Jl Daeng. Tata Raya, Makassar 90224. Indonesia 2) Indonesian Cereals Research Institute (ICERI). Jl. Dr. Ratulangi, Maros. 90514. Indonesia 3) Graduate School of Horticulture, Chiba University. Matsudo, Chiba 271-8510. Japan

Abstract : The aim of research is to determine the influence of urea granulated zeolite and nitrification inhibitor on the growth maize (Zea mays L. Var. B8). The layout of study was using completely randomized design with with 3 replications. Treatments were namely urea granule, urea granulated zeolite, urea granulated zeolite neem, urea granulated zeolite dicyandiamide, urea granulated dicyandiamide, urea granulated neem and a control. Fertilization was done in split, 8 days and 29 days after planting with total nitrogen application was 150 kgN ha1. The parameters growth were include height of plant stem (cm), leaf chlorophyll content ( weight of 5 corncobs (gram) and plant dry weight (kg). The data of growth were analysed by using the F-test analysis to find differences. The results of study show that urea granulated with zeolite and nitrification inhibitors affected the growth of maize (Zea mays L. Var. B8). Keywords: Urea, zeolite, neem, dicyandiamide, nitogen, growth of maize(Zea Mays L.) 1. Introduction Maize is one of the world's most important cereal crops besides rice andwheat. Corn plant has long been known by the people of Indonesia and SouthSulawesi province is one of the centers of national maize development located ineastern Indonesia. Corn is also a valuable economic commodity because of hisposition as the main source of carbohydrate after rice. Corn is widely used by thecommunity as a staple food, vegetables, processed into snacks, as feed forlivestock, and many other benefits. Therefore, the demand of corn is increasingfrom year to year and the efforts have been made by Indonesian farmers toincrease the productivity of the corn crop by maintenance of soil fertility. The common practice by farmer to maintenance soil fertility is applyingfertilizer either organic or synthetic one. Provision of fertilizer on land can bedone either before or after planting. The regular fertilizers has used by farmers isurea as nitrogen source. According Sharma and Prasad (1996) nitrogen is anelement of main nutrients for plant growth which in generaly indispensable for theformation or growth of vegetative parts of the plant, such as leaves, stems, androots, as well as flowering and fruiting plants. However, urea is a volatile fertilizer and easily leached by water as nitrate (NO3-) so that addition treatment isnecessary to prevent that process. One way to do is through the granulationprocess, namely the formation of powder particles become larger (multiparticle).

Another way to increase the efficiency of nitrogen contained in the urea is to add material to slow release such as zeolite into the urea fertilizer. According to Treacy and Newsam (1988) zeolites are aluminosilicate minerals that have a tri dimentional structure that hollow and aisle, hence its have spending surface area that lead binding nitrogen. Another, way to improve the efficiency of fertilizer beside use of zeolite can also be done by inhibiting the nitrification process. Inhibition of nitrification can be utilized by addition of materials nitrification inhibitors such AM (2-amino 4-chloro 6-methyl pyrimidine), ST (2-sulfanilamide thiazone), DCS (N-2, 5-dichlorophenyl succsinamic acid) dan ASU (1-amino 2 thiourea). However, the common kind of nitrification inhibitor that available in market are DCD (dicyandiamide), nitrapiryn dan CCC (wax-coated calcium carbide) (Mosier, 1996). Considering the price of synthetic nitrification inhibitor is expensive, invention of nitrification inhibitors that easy to find in Indonesia and cheap is necessary. One of organic material that has capacity to inhibit the nitrification process is seeds of neem (Azadirachta indica) (Hala et al, 2014). This is consistent with the results of Akhtar (2000): Majumdar et al (2004) which showed that the neem seeds serve as a nitrification inhibitor. The working principle of nitrification inhibitors is to inhibit the process of conversion of ammonium to nitrate and can improve the efficiency of nitrogen use by plant. Inhibition of the conversion of ammonium to nitrate in the soil will also provide an opportunity for the plant to absorb more ammonium as well as nitrate for growth (Malla et al, 2005).

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Based on the description outlined above it is clear that applying urea in cooperated with the zeolite and nitrification inhibitor in granular type seems has potential to reduce application of nitrogen fertilizer (urea) and would lead efficiency, reducing cost production and also less pollution to air and water Malla et. al, 2005; Jumadi et al, 2008). However, rare research has been done to assess the effect of combination zeolite as slow release and neem to maize growth. Hence, the aims of current studies are to determine the effect of urea granulation with zeolite and nitrification inhibitor (neem) tothe growth of maize plants. 2. Research Methods The field experiment location was in Balitsereal Indonesian CerealsResearch Institute (ICERI) Maros district, South Sulawesi province, Indonesia(4°59'11.3"S 119°34'34"E). The variety of Maize (Zea mays L.) was Var. B8 andthe treatments consist of control (K), urea (U), urea zeolite (UZ), neem urea (UN),urea dicyandiamide (UD), neem zeolite urea (UZN), and urea zeolitedicyandiamide (UZD) with three replicates. Fertilizers were granulated usinginclined pan granulator which contained around 45-47% nitrogen. Landpreparation followed the practices of local farmers to allow a better analysis of thedata on a regional scale. The Fertilization was done in split time, first fertilizationwas performed at 8 days after seeding with fertilizer amount as 100 kg N/ha aswell as addition KCl as much as 100 kg/ha. The second fertilization was done 29days after planting with the same amount of fertilizer at first and also superphosphate (SP3) as much as 100 kg/ha. Plants were watered regularly andharvesting at 92 days after planting. Harvesting is done by first picking fruit corn,peeling skin, then give the number on each corn. Meanwhile, corn stalks atharvest to plant dry weight measurement. The growth rate parameters wereobserved in this study including plant height (cm), leaf chlorophyll content (%),five cob weight (grams), and plant dry weight (grams). Means and standarddeviations of the data were calculated.

Means were compared and subjected to thetest Significant Difference Distance Duncan (P< 0.05) using SPSS software(Ver.21.0 for windows, SPSS Inc., Chicago, USA). 3. Result And Discussion 3.1 Plant height and chlorophyl content The plant height was observed every week during plantation time. Table 1shows that the average data of corn growth, Treatment control (K) has lowest andsignificantly different from the other treatments. At 50 day after plantationshowed an average growth of control (K) significantly different from the six othertreatments (U, UZN, UZD, UD, UZ, and UN). Well know that less nutrient in soilwill affected the growth of plant, particularly the amount of nitrogen where is oneof important element need to plant.Nitrogen is a nutrient that can help thevegetative growth of plants, including highgrowth plants. Well growth of plantindicates the absorption of nutrients in the soil to the fullest. According Sitompuland Guritno (1995) nitrogen is a major nutrient for plant growth, which in generalis indispensable for the formation or growth of vegetative parts of the plant, suchas leaves, stems, and roots. In Table 1 shows that the growth of the corn plant height measured from 8day after plantation has the same relative growth for all treatment up to day 29.The increase in plant height starts to seems different from the measurements madeon day of 36. This difference continues to increase until 50 day. Presumably thatwere caused by the absorption of nutrients by the roots, where the fertilizer isgiven in the form of urea was not directly absorbed by the corn plant. Thetransformation of applying fertilizer was first being recast into a new ammoniumform that can then be absorbed by plants and by nitrification that form into nitrate.Therefore, addition of nitrification inhibotir and zeolite as slow release of nitrogenwill lead retention amount of ammonium as well as nitrate in soil then plant willabsorb nitrogen efficiently.

Table 1. Average data growth of corn plant height (cm) every week.

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Jumadi et al (2008) have also pointed that nitrogen required by the plantthroughout the growing corn. At the beginning of the accumulation of N in plantgrowth is relatively slow and after 4-week-old crop N accumulation takes placevery quickly. At the time of flowering (male flowers appear) maize crop has beenabsorbing N as much as 50 % of all its needs. Sharma and Prasad (1996)suggested that urea is a dry material in the form of granules or bulk, urea-N is rapidly hydrolyzed to NH4+. This fertilizer is often used for direct application in amixture of fertilizers and nitrogen in solution. The application of nitrogen in theform of a urea-N, and approximately 66 % of urea-N hydrolyzed into ammonia-N. Effect of fertilizer to chlorophyll content can be see clearly at table 2 and3. In Table 2 show that the content of chlorophyll in Cob leaf plant significantlyless in control, while no differences in U, UZ, UZD, UD and UN, but UZN haslower chlorophyll content than UZ. Chlorophyll is the main component in theprocess of photosynthesis that provide essential nutrition for enhance the growthrate of plant. In Table 3 shows that treatment of control was content lesschlorophyll flag leaf and significantly different from the other treatments (U,UZN, UZD, UD, UZ, and UN), while plants have leaf chlorophyll content flagwas highest at UZ treatment, but not significantly different from the flag leafchlorophyll content of corn plants treated with U, UZN, UZD, and UD. It is appearing that the form of urea granulation treatment with zeoliteand nitrification inhibitors has positive affect to chlorophyll content of plants,both leaf chlorophyll content and leaf flag cob corn crop. These results also wellexplained that it might be due to the difference in nutrients in soil, especiallynitrogen absorbed by each plant corn. Table 2. Average Data Cob Leaf Chlorophyll Content of Plants (%) Treatment

Chlorophyll Content (%)

K U UZ UZN UZD UD UN

39.6867a 52.4893bc 54.8200c 46.9733b 49.9200bc 49.7667bc 48.5400bc

Different letters refer to significant differences Duncan test (P < 0.05).

Increased levels of chlorophyll show that inorganic nitrogen fertilizer (urea) that is given can be absorbed by plant roots and used to form more chlorophyll. This is as disclosed Sitompul and Guritno (1995), nitrogen is one of the main

constituent component of leaf chlorophyll which is about 60%, and act as enzymes and membrane proteins. Fathan et al (1998) prompted nitrogen in the body of plants found in the form of inorganic joining to elements C, H, and O to form amino acids, enzymes, nucleic acids, and chlorophyll. Thus increasing the rate of photosynthesis and more produce asimiliat substance. Table 3. Average Data Flag Leaf Chlorophyll Content of Plants (%) Treatment

Chlorophyll Content (%)

K U UZ UZN UZD UD UN

38.2400a 52.4667bc 53.6400c 47.0533bc 50.0667bc 49.2000bc 45.7267b

Different letters refer to significant differences Duncan test (P < 0.05).

3.2 Maize Cobs Weight Urea granulation with zeolite and nitrification inhibitors affects the cropyields of corn. In table 4 shown that the difference between the five cobs of corn plants treated with control to other treatments. Weight of five cobs at control treatment had lowest among all treatments. The low weight of five cobs of corn in the control treatment that can be due to the lack of nitrogen was absorbed by plants. This is also correlated with Buckman and Brady (1982) that nitrogen fertilizer has pronounced in plants to increase growth rate and gives a green color in the leaves, hence maize enhance the cobs (fruit) as well as increase the protein content. Further, state by Mosier et al (2000), that the lack of nitrogen in the plants will have symptoms of yellow color in the leaves, seeds and decrease of fruit weight. Table 4. Average Data Five Weight Corn Cobs (gram) Treatments Weight (gram) K U UZ UZN UZD UD UN

754.0000a 1420.6667c 1270.6667b 1302.6667bc 1394.0000bc 1406.6667c 1374.0000bc

Different letters refer to significant differences Duncan test (P < 0.05).

According to Sharma and Prasad (1996), long and weight of maize cobs is produced by plants by the capacity of existing photosynthesis in plants.

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Nitrogen is a very important nutrient in plant growth. Nitrogen is used to give a dark green color in the leaves as a component of chlorophyll, stimulating rapid growth, and increased plant height, number of tillers, leaf size, and the protein content in the seed (Prasad and Power, 1995). In addition, another factor that plays a role in the process of photosynthesis is the sunlight received by plants. The process of photosynthesis can take place optimally since the reception of light by plants takes place optimally. Beside of nitrogen, plants also need the element potassium and phosphorus for growth well. Potassium in plant physiological processes and functions controlling cell metabolism, increase resistance to disease. Potassium nutrient deficiencies cause weak of stalk, the transport of nutrients, impaired respiration and photosynthesis, which in turn reduces the production, while the role of phosphorus for plants is accelerating the maturation of plants, play a role in the formation of flowers and seeds, offsetting the effect of excess nitrogen, helps the development of roots and root hairs, as well as strengthening the stem. 3.3 Dry weight maize Corn plant dry weight data obtained after the corn crop dried. Data dry weight of corn plants can be seen in Table 5 below: Table 5. Average Data Dry Weight Maize (Kg) Treatment Weight (Kg) K U UZ UZN UZD UD UN

0.6867a 1.2567b 1.0200ab 1.1000b 1.1400b 1.2033b 1.3333b

Different letters refer to significant differences Duncan test (P < 0.05)

Plant growth is marked by the increasing size and weight of dried plants. This increase is caused by the increase in plant organs such as plant height, leaf number, and leaf area, as a result of plant metabolism is influenced by environmental factors such as sun around the plant, temperature, water and plant nutrients. The appearance of a plant is the result of an interaction between genetic and environmental factors. Each plant varieties have different capabilities in terms of utilizing the means to grow and the ability to adapt to the surrounding environment, so that the potential outcome is also different. The amount of the dry weight of the plant is closely related to organ leaves on the plant, because the leaf photosynthesis process occurs in which the results

will affect the photosynthesis of the plant dry weight. Based on the research conducted, the treatment given very significant effect on plant dry weight of corn crops have been harvested with the greatest amount of the dry weight of corn plants treated U, UZN, UZD, UD, and the UN. As explained earlier that nitrogen levels affect the number of leaves and leaf area of plants, thus indirectly maximize photosynthesis. Photosynthesis results are then accumulated into a corn plant dry weight. According Martajaya (2002), when the plants get enough N then the leaves will grow bigger and expand its surface. Wider leaf surface allows it to absorb a lot of sunlight so that the process of photosynthesis also takes place more quickly, consequently fotosintan formed will accumulate on the dry weight of the plant. 4. Conclussion We conclude that applying of urea granulation with zeolite and nitrification inhibitors enhance growth of maize (Zea mays L. Var. B8). Bibliography [1] Akhtar M. 2000. Nematical potential of the neem tree Azardirachta indica (A.Juss). Integ. Pest Manag. Rev. 5: 57 – 66. [2] Abdillah Agung. 2008. Pengaruh Zeolit dan Piupik K Terhadap Ketersediaan dan Serapan K Tanaman Padi di Lahan Pasir Pantai Kulonprogo. Skipsi. Fakultas Pertanian Universitas Sebelas Maret. Surakarta. [3] Bangun, B.A.B., Jasmani,G. dan Ferry, E.S. 2014. Respon Pertumbuhan dan Produksi Sawi (Brassica sinensis L.) By Giving Mineral Zeolite and Nitrogen. Jurnal Online Agroekoteknologi. ISSN No. 2337-6597 Vol.2, No.3 : 12271230. [4] Buckman, H.O. dan Brady, N.C. 1982. ilmu Tanah. (Terjemahan: Soegiman). Jakarta: Bharata Karya Aksara. [5] Fathan, R. M., Raharjo, A.K., dan Makarim. 1998. Hara tanaman jagung. Bogor: Puslitbangtan. [6] Hala Y, Jumadi O, Muis A, Hartati, Inubushi K. 2014. Development of Urea Coated with Neem (Azadirachta indica) to Increase Fertilizer Efficiency and Reduce Greenhouse Gases Emission. J.Teknology, UTM, Malaysia. [7] Jumadi O. Hala Y, Inubushi K. 2005. Production and emission of nitrous oxide and responsible microorganisms in upland acid soil in Indonesia. Soil Science and Plant Nutrient. 51 (5): 693-696. [8] Jumadi O, Hala Y, Muis A, Ali A, Palennari M, Yagi K, Inubushi K. 2008a. Influences of chemical fertilizers and a nitrification inhibitor

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[9]

[10]

[11]

[12]

[13]

[14]

[15]

[16]

on greenhouse gas fluxes in a Corn (Zea mays L.) field in Indonesia. Microbes Environ. 23: 29-34. Majumdar D, Pathak H, Kumar S, Jain MC.2002. Nitrous oxide emission from a sandy loam inceptisol under irrigated wheat in India as influenced by different nitrification inhibitors. Agri. Ecosys. Environ. 91: 283-293. Majumdar D, Pandya, Arora A, Dhara S. 2004. Potential use of Karanjin (3Methoxy Furano2‟,3‟,7,8 Flavone). Arch. Agron and Soil Sci. 55: 455 – 465. Malla G, Bhatia A, Pathak H, Prasad S, Jain N, Singh J. 2005. Mitigating nitrous oxide and methane emission from soil in rice-wheat system of the IndoGangetic plain with nitrification and urease inhibitors. Chemosphere 58: 141147. Mosier AR and Kroeze C. 2000. Potential impact on the global atmospheric N2O budget of the increased nitrogen input required to meet future global food demands. ChemosphereGlobal Change Sci. 2:465-473. Prasad R and Power JF 1995. Nitrification inhibitor for agriculture, health and the environment. Adv. Agron 54:233-281. Sitompul, S.M. dan B. Guritno. 1995. Analisis Pertumbuhan Tanaman. Yogyakarta: UGM press. Sharma SN and Prasad R. 1996. Use of nitrification inhibitor (neem and DCD) to increase N efficiency in maize-wheat cropping system. Fertilizer research 44:169 – 175. Treacy MMJ and Newsam JM. 1988. Two new three dimensional twelve ring zeolite frameworks of which zeolite beta is a disordered intergrowth. Nature 332, 249-251.

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FOOD and FEED TECHNOLOGY

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Carcass Quality, Blood Profile and Organ Histopathology of Sheep Fed Organic Additive Contaning Probiotic and Micromineral Enriched Yeast Ade Erma Suryani, Lusty Istiqomah, Ahmad Sofyan, Awistaros Angger Sakti andMohammad Faiz Karimy Division of Feed and Animal Nutrition, Research Unit for Processes Development and Chemical Engineering (BPPTK) – LIPI Jl. Jogja-Wonosari Km. 31,5, Gading, Playen, Gunungkidul, Yogyakarta 55861 email: [email protected] Abstract:The aim of the study was to evaluate the effect of combination of probiotics (Pediococcus acidilactici)and micro mineral enriched yeast (MEY) administration on internal organ and carcass quality andhaematological profile and liver histopathology of local sheep. The experiment was conducted using sixlocal sheep (body weight 13 •} 0.8 kg) were arranged in a complex randomized design with 2 treatment 3replication and each replication consisted of one sheep. Sheep was randomly placed in individual cagesand given one of two treatments were P0 (control, basal diet without ProMEY), P1 (basal diet+ProMEY).Basal diet consisted of forage and concentrate (60:40 in dry matter basis and given twice a day.Concentrate feed was given after animal consumed forages in the morning. Feed additive of ProMEY (15g/d/head) was added into concentrate. Water were provided ad libitum all over the experimental period.At 10 weeks of feeding trials, one sheep was randomly selected from each treatment and slaughtered toevaluate the carcass quality. Analysis of hemoglobin (Hb), packed cell volume (PCV), leucocyte,thrombocyte, basophil, neutrophil, and eosinophil, were analyzed by microhematocrit. Histopathologyprofile was evaluated by lesion degree of organ. The result indicated a significanly different betwencontrol treatment (P0) and ProMEY treatment (P1) on performance of liver. In addition, sheep fed byProMEY administration produced higher number of erythrocytes (26%), hemoglobin (25%), PCV (15%),lymphocyte (11.5%) and monocyte (50%) than control. Histopathologal profile result showed that therewas no alteration changes in liver. It could be concluded that combination of Probiotics and MicromineralEnriched Yeast could improved the health status of sheep. Keywords : probiotics, micro mineral, carcass quality, haematological. 1. Introduction Small ruminants has economic and ecological values that are important in farming systems throughout the developing world (Davendra., 2005). Improvement of feed management is one important factor in the development of small ruminant livestock. Animal feed supply consists of continuity in quality and also quantity still becomes major challenge for Indonesian farmers. Diet composition and probiotics administration are known could affect the performance of ruminant. Probiotics administration could increase feed intake. Several studies about probiotics mentioned that the probiotics increased dry matter intake and improve the ability of the fiber degradation. Due to the improvement of cellulolytic bacteria in rumen (Khalid et al. 2011). Alternatives of feed additive such as dietary acidifiers, essential oils, probiotics and prebiotics had been introduced to improved animals performance (Mohammed et al., 2013). Probiotics are beneficial live microorganisms that can be added to feed or water as single and mixed cultures (Todorov et al., 2007). In sheep, administration of probiotics in feed can directly affect in preventing and combating the pathological conditions that arise due to an imbalance in the digestive tract. The administration of probiotics and prebiotics help in correcting the imbalance of bacteria, provides energy and help in rehydration to

reduce recovery time from stress or disease treatment in sheep (Nicole et al., 2010). In addition, administration of probiotics affect feed consumption, daily gain and absorption of nutrients (Chiofalo et al., 2004) and thus reduce the mortality and accelerated weaning of young animals (Khalid et al., 2011). Minerals were required in relatively small quantities compared with other feed substances; however its deficiency was very influential on livestock performance (Herdian et al., 2013). Effect of many trace minerals absorption and dietary factors that affect bioavailability of mineral differ greatly between ruminants. In ruminant, microbial digestion in the rumen and reticulum precedes mammalian digestion in the abomasum and small intestine (Spears, 2003). A number of studies showed that the administration of minerals in organics form increased its bioavailability and improved the performance of livestock. The administration of probiotics in feed intended to create a balance of beneficial microorganisms in the process of degradation of nutrient components in rumen. Enzymatic activity against degradation of fiber component could be increased if the production of fiber-digesting enzyme increased. Therefore the availability of the required elements as a factor of

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microbial growth becomes very important. Some micromineral especially Zn played an important role in encouraging the enzyme activity in the rumen and it became required element in the rumen media as growth factors of microbe (Krisnan et al., 2009). Minerals in organic compounds form were easily utilized by the body and non toxic. Organic minerals could be synthesized by microorganisms help through biofermentation process. Mineral requirements for animals were highly dependent on the level of productivity (Lee, 1996). Mineral were important element that required in small amount by all animals. These minerals performed essential functions in the body and directly involved in their productive performance (Teixeira et al., 2013). Therefore, the availability of the required elements as microbial growth factor becomes very important. From this approach, it appeared an idea to combine the administration of probiotics with micro mineral enriched with yeast and its influence on internal organ and carcass quality, haematological profile and liver histopathology of local sheep. 2. Methods and materials 2.1 Probiotic and organic mineral preparation Probiotics consisted of lactic acid bacteria (Pediococcus acidilactici). The LAB was isolated from rumen. Isolates were cultivated in deMann Rogosa Sharpe Broth / MRSB medium for 18 h at 37 °C and dried cultured prepared by Spray Drying Method as previously described by Sofyan et al. (2015). The dried culture colonies was evaluated by spread plate method and prepared to reach 109 cfu/g of minimum colonies. Preparation of micromineral enriched yeast (MEY) was performed using fermentation processes using yeast S. cerevisiae ATCC 9763 as inoculum. Yeast were inoculated into medium contained of Fe, Mn, Cu, Co, Zn, and I (Sofyan et al., 2015). One kg of cassava flour meal supplemented with mineral solution contained FeCl2.4H2O (0.177 g), MnCl2.4H2O (7.129 g), CuSO4.5H2O (9.810 g), ZnSO4.7H2O (12.646 g), CoCl2.6H2O (0.192 g), and KI (0.217 g) was used as fermentation medium. Then, the inoculated medium was incubated for 7 days in facultative fermentation condition MEY was harvested and dried by oven at 55°C (up to 24-48 h, DM 10%), then ground and sieved to obtain a homogenous particle size of 1 mm. 2.2 Animal and feed Six local sheep were adapted by basal diet feed and medicated by antibiotics for eliminate the parasites. The experiment was conducted using six local sheep (body weight 13 •} 0.8 kg) were arranged in a complex randomized design with 2 treatment 3 replication and each replication consisted of one sheep. Sheep was randomly placed in individual

cages and given one of two treatments were P0 (control, basal diest without ProMEY), P1 (basal diet+ProMEY). Basal diet was formulated according to nutrient requirement for sheep (NRC, 2007) consisted of forage and concentrate (60:40 in DM basis) and given twice a day. Corn stover (milk dough stage) was chopped as forage diet. Concentrate was composed from rice bran (31.0%), pollard (66.67%), urea (0.35%), calcium carbonate (1.0%), and molasses (0.5%). Nutrient composition of basal diet was shown in Table 1. Concentrate feed was given after animal consumed forages in the morning. Feed additive of ProMEY (15 g/d/head) was added into concentrate. Water were provided ad libitum all over the experimental period. Table 1. Composition and nutrient content (DM basis) on the basal diets used in the experiment Nutrients Total Digestible Nutrients (TDN) Crude Protein (CP) Ether Extract (EE) Crude Fiber (CF) Calcium (Ca) Phosporus (total P) Sodium (Na) Cloride (Cl) Potassium (K) Sulfur (S) Cobalt (Co) Cuprum (Cu) Magnesium (Mg) Selenium (Se) Zinc (Zn) Iron (Fe) Manganese (Mn)

Unit (%) (%) (%) (%) (%) (%) (%) (%) (%) (%) (ppm) (ppm) (ppm) (ppm) (ppm) (ppm) (ppm)

Content 62.91 11.28 3.55 21.17 0.36 0.34 0.01 0.02 0.46 0.05 0.03 1.62 0.18 0.11 0.16 0.26 0.01

2.3 Evaluation of internal organ and carcass quality At 10 weeks of feeding trials, one sheep was randomly selected from each treatment and slaughtered to evaluate the carcass quality. Carcass part of longissimus dorsi were ground for the further analysis. Meat cholesterol content was analyzed using colorimetric methods according to Liebermann- Burchad reaction as described by AlBalaa et al. (2014), while the fatty acids composition was analyzed using gas chromatography (Kišidayova et al. 2014). Measurement of pH and moisture content was referred to AOAC (2012). Cooking loss was determined according to Kondjoyan et al. (2013). Analysis of water holding capacity analysis was determined mehods as described by Li et al. (2013). Meat colour was measured by capturing the picture using digital camera (Exlim Casio 12.0 Mpixels) and interpreted descriptively.

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2.4 Haematological analysis.

and

histopathological

Blood samples from jugular vein were individually collected into 10 ml lithium heparin vacuum tubes (Agazzi et al., 2014) consisted of 10 uL EDTA (ethylene diamanine tetraactic acid). Analysis of hemoglobin (Hb), peak cell volume (PCV), leucocyte, thrombocyte, basophil, eosinophil, neutrophil were analyzed by microhematocrit methods according to Bain (2014). One mL of blood and prolonged during 30 min to 1 hour centrifugation in graduated glass tubes with a constants internal rotation. A small volume of blood is was taken by capillary into an ungraduated capillary and leaved about 5 mm unfilled. The end of the tubes distant from the column of blood was sealed by heat. Centrifugation was conducted for 10 minutes at high speed 15.000 xg to separate column of the blood into red cells, buffer coat and plasma. Tissue from liver was fixed in formalin solution (10%). Sample was stained with hematoxylineosin

(HE) according to Elliot (2012). Histopatology profile was evaluated by lesion degree of organ as described by Povoa et al. (2014). The stained sections were visualized and recorded using the digital camera connected to electronic microscope with 200 X magnification. 2.5 Data analysis The effect of treatments on sheep internal organ and carcass quality, physical and chemical characteristic of carcass haematological profile and liver histopatology were evaluated by descriptive analysis. 3. Results and discussion 3.1 Internal organ and carcass quality Performance of sheep in control treatment (P0) and ProMEY treatment (P1) showed that there were significantly different among treatments (Table 2). Physical and chemical characteristic of carcass from sheep could be seen in Table 3.

Table 2. Performance, internal organ and carcass percentage of sheep administered by probiotic (Pro) and micromineral enriched yeast (MEY)

Level of oleic acid observed for the ProMEY treatments higher than others. could be attributed to the fatty acid transfer from blood to muscle involving stearoyl-CoA desaturase. Ferreira et al. (2014) revealed that stearoyl-CoA desaturase contributed to synthesis of fatty acids. This result was supported by Santillo et al. (2012) that meat lamb receiving milk replacer consisted of probiotic Lactobacillus and Bifidocateria had 15% lower concentration of cholesterol than control. The similar result also found in profile of polyunsaturted fatty acids. Supplementation of probiotic increased fatty acid composition linoleic (18:2) and oleic acid (18:1). In recent study, some of fatty acid could not

be detected, it might be related to either the lower content or destructed compound during preparation. Beside chemical properties, meat colour was necessary to be evaluated (Figure 4). Meat colour could be associated by intramuscular fat and marbling score (Li et al. 2013) and indicated of myoglobin (Roldan et al. 2013). Meat taken from longisimuss dorsi from sheep fed by ProMEY showed moreredness color than control. Although it should be further analysis, redness color of meat related to profile of blood. Sheep treated by ProMEY had higher numerically value of red blood cell than control (Table 4). In other animal, Zheng et

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al. (2014) reported that meat of broiler supplemented by probiotic produced better meat color parameters consisted of lightness, redness and yellowness although the mechanism of probiotic (Enterococcus faecium) effect was still unclear

described. Redness color of raw meat is became important parameter to be considered in meat quality evaluation and implied the economical value of meat.

Table 3. Physical and chemical characteristic of carcass meat from sheep administered by probiotic (Pro)and micromineral enriched yeast (MEY).

Figure 1. Texture of longisimus dorsi from sheep administered by probiotic (Pro) and micromineralenriched yeast (MEY) (A: Control; B: ProMEY) 3.2 Haematological histopathology

profile

and

liver

Health status of sheep was distinguished by evaluation of blood profile and liver histopathology. Blood profile consisted of erythrocytes, leukocytes, hemoglobin, packed cell volume/PCV, total plasma protein, neutrophil, eosinophil, basophils, lymphocytes, and monocytes. This blood profile of sheep from both treatments were described in Table 5 and liver histopathology was visualized on Figure 2.

Figure 2. Histopathologal profile of liver from sheep administered by probiotic (Pro) and micromineralenriched yeast (MEY) Histopathologal profile result showed that there was no alteration changes in liver. In addition, sheep fed by diet containing administration of organic additive ProMEY higher than control in number of erythrocytes (26%), hemoglobin (25%), PCV (15%), lymphocyte (11,5%) and monocyte (50%). However, the P1 produced lower in leukocytes (eosinophil) and similar in TPP (neutrophil) than control. Šimpraga et al. (2013) reported that blood profile in sheep consisted of erythrocytes, leukocytes, hemoglobin in lowering limit were 7.81x106/uL, 3.02x103/uL and 7.81 g/dL respectively. In addition, blood parameters were affected by environmental condition including nutrient composition of feed. Leukocyte was seen to similar, while erythrocyte and hemoglobin were higher than control. It was indicate that feed additive of ProMEY had positive effect to support health status of sheep. Moreover, Agazzi et al. (2014) reported that calf 6.33 vs 7.00 x 106/uL erythrocytes, leukocytes 9.42 vs 8.53 x 103/uL,monocytes 17.0 vs 17.4%, lymphocytes 38.3 vs 40.9%, basophils 0.16 vs 0.21%, neutrophils 42.7 vs 38.8%, the different result betwen calf with administration specific probiotics in basal diet than calf with fed basal diet. The results showed similar response of biochemical parameters with this study. It seems that administration of ProMEY indicated improvement of immunity parameters. In contrary, Gordon et al. (2013) reported that chelated mineral consisted of Cu, Zn, Mn and Co produced no different effect on

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digestibility. Improvement of bioavailability of Selenium in organic form supported its functions as an immunomodulator and antioxidant in selenoproteins such as glutathione peroxidase. Dietary probiotic from lactic acid bacteria enhance digestive process and immune function. Probiotic could stimulate lymphocite responsisiveness in sustaining cell-mediated and humoral immunity of lambs. Immune-regulatory function enhancement

was indiacted by increasing number of interleukin10 as an anti-inflamantory cytokine (Harden et al. 2013). Intramuscular fatty acid composition was affacted by biohidrogenation in rumen. Although the histopatological organ of liver was similar in both treatments, based on the performance parameter of treated sheep fed by ProMEY resulted better health status than control.

Table 4. Blood profile of sheep administered by probiotic (Pro) and micromineral enriched yeast (MEY)

References

[4] Ferreira, E.M., Pires, A.V., Susin, I., Gentil, R.S., Parente, M.O.M., Nolli, C.P., Meneghini, R.C.M., Mendes, C.Q., and Ribeiro, C. V. D. M. (2014). Growth, feed intake, carcass characteristics, and meat fatty acid profile of lambs fed soybean oil partially replaced by fish oil blend. Animal Feed Science and Technology, 187(3-4), 9-18.

[1] Agazzi, A., E. Tirloni, S. Stella, S. Maroccolo, B. Ripamonti, C. Bersani, J. M. Caputo, V. Dellorto, N. Rota, and G. Savoini (2014), Effect of species-specific probiotics addition to milk replacer on calf health and performance during the first month of life. Ann. Anim. Sci. 14 (1) : 101-115

[5] Harden, L. M., Rummel, C., Luheshi, G. N., Poole, S., Gerstberger, R., & Roth, J. (2013).Interleukin-10 modulates the synthesis of inflammatory mediators in the sensorycircumventricular organs: implications for the regulation of fever and sickness behaviors. JNeuroinflammation, 10, 22.

[2] Chifalo, V., L, Liotta and B. Chiofalo. 2004. Effect of administration of lactobacilli on body growth and on the metabolic profile in growing Maltese goat kids. Reproduction Nutrition Development., 44 : 449-457.

[6] Herdian H., A. Sofyan., A.A. Sakti., H. Julendra., M. F. Karimy., A.E. Suryani., E. Damayanti dan L. istiqomah. 2013. Media Peternakan, Desember 2013, pp. 203-208. ISSN 0126-0472 EISSN 2087-4634. DOI: 10.5398/medpet.2013.36.3.203

4. Conclusion Administration of probiotics (Pro) (15 g/head/day) contained Pediococcus acidilactici (1x109 cfu/g) that combined with micromineral enriched yeast (ProMEY) seems to increase precentage of carcass quality and improve the health status of sheep.

[3] Davendra C. 2005. Small Ruminant in Asia; Contribution to food security, property alleviation and opportunities for productivity enhancement. Proceeding of international workshop on small ruminant production and development in Shout East Asia. MEKARN. Nong Lam, HCMC, Vietnam.

[7] Khalid, M. F.., Shuhzad, M. A., Sarwar, M., Rehman, A.U., Sharif, M. and N. Mukhtar. 2011. Probiotics and Lamb Performance: A Review. African Journal of Agricultural Research. 6 (23): 5198-5203. [8] Krisnan R., B. Haryanto., and K.G. Wiryawan. 2009. The Effect of Combined Probiotics with

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Catalyst Supplementation on Digestion and Characteristic in Priangan Sheep. JITV 14 (4): 262-269.

[18] Todorov, N., I. Krachusnov, A. Alexandrov and D. Djuvinov. 2007. Guide on Animal Nutrition. Matkom, Sofia.

[9] Lee R. McDowell. 1996. Feeding Minerals to Cattle on Pasture. Animal Feed Science Technology 60 (1996) 247-271

[19] Zheng, A., Luo, J., Meng, K., Li, J., Zhang, S., Li, K., Liu, G., Cai, H., Bryden, W.L. and Yao, B. (2014). Proteome changes underpin improved meat quality and yield of chickens (Gallusgallus) fed the probiotic Enterococcus faecium. BMC Genomics, 15(1), 116

[10] Li, X., Ekerljung, M., Lundstrom, K., and Lunden, A. (2013). Association of polymorphisms at DGAT1, leptin, SCD1, CAPN1 and CAST genes with color, marbling and water holding capacity in meat from beef cattle populations in Sweden. Meat science, 94(2), 153-158. [11] Mohammed HH, El-Sayed BM and Ali MA. 2013. Effect of Commercial Feed Additives on Performance, Economic Efficiency, Blood Metabolites and Some Maintenance Behaviour in Goats. Journal of Veterinary Science and Medical Diagnostic 2:2. doi: http://dx.doi.org/10.4172/2325-9590.1000112 [12] Nicolae Corcionivoschi, Dan Drinceanu, Ioan Mircea Pop, Deirdre Stack, Lavinia Ster, Callin Julean and Billy Bourke. 2010. The Effect of Probiotics on Animal Health. Scientific Papers: Animal Science and Biotechnologies. 43 (1) [13] Roldan, M., Antequera, T., Martin, A., Mayoral, A. I., & Ruiz, J. (2013). Effect of different temperature–time combinations on physicochemical, microbiological, textural and structural features of sous-vide cooked lamb loins. Meat science, 93(3), 572-578. [14] Santillo, A., Annicchiarico, G., Caroprese, M., Marino, R., Sevi, A., and Albenzio, M. (2012). Probiotics in milk replacer influence lamb immune function and meat quality. Animal, 6(2), 339-345. [15] Sofyan A, Sakti AA, Karimy MF, Julendra H, Istiqomah L, Herdian H, Damayanti E, Suryani AE. 2015. Effectivity of probiotic, micromineral enriched yeast and their combination with neem (Azadirachta indica) leaves containing tannin on fermentability and digestibility of king grass (Pennisetum hybrid) assessed by in vitro gas production technique. JITV. (Under Review) [16] Spears, J.W. 2003. Trace Mineral Bioavailability in Ruminants. J. Nutr. 133:1506S-1509S. [17] Teixeira et al. 2013. Mineral Requirement for Growth of wool and Hair Lambs. Revista Brasileira de Zootecnia Vol. 42. No. 5 p : 347353. ISSN 1806-9290.

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Influence of Flouring Method on Characteristic of Tacca Flour: Phytochemical, Chemical and Resistant Starch Analysis Miftakhussolikhah, Dini Ariani, Tri Wiyono UPT. BPPTK of Indonesian Institute of Science Jl. Yogya-Wonosari km 31,5 Gading, Playen, Gunungkidul, DIY, Indonesia e-mail: [email protected] Abstract: Tacca (Tacca leontopetaloides) is a tuber that has high carbohydrate content, but itspotential has not been widely used and studied. Tacca sp. grows scatteredly in high-salinityareas such as southern coast of Java. Commonly, Tacca flour is utilized as raw material offood product e.g. noodle, and furthermore in recent studies it can be used aspharmaceutical excipient. The goal of this study is to compare the characteristics of Taccaflour by differentiating the flouring method. Tacca flours were made by slicing (chippedtacca flour) and grating method (pressed tacca flour). Phytochemical, chemicalcomposition and resistant starch content of both were analyzed and compared each other.The results showed that there were no significant difference of water, fat and carbohydratecontent between chipped and pressed Tacca flour as well as its phytochemical content. Anumber of terpenoid TLC spot qualitatively apeared thicker on pressed flour rather thanchipped one. Ash, protein, amylose, and resistant starch content of chipped Tacca flourwas higher than that of pressed Tacca flour. Keywords: tacca, sodium metabisulfite, starch, flour, noodle, phytochemical 1. Introduction Tacca (Tacca leontopetaloides) or jalawure or Polynesian tacca is a plant that grows wildly in coastal areas with high-salinity e.g. at southern coast of West Java and open location with high sunlight exposure like grassland and savannas. It can be reproduced vegetatively (by bulbs) or generatively (from seed). Commonly, tacca utilization as food is still limited as raw material for traditional cakes. Based on tacca potential as carbohydrate source, chemical properties of arrowroot flour need to be determined, so it can be developed as functional food. Processing tuber into flour could increase tuber shelf life and economic value (Richana and Sunarti, 2004). There were two flouring method that usually apllied, chipping method and pressing method. Pressing method could optimize time and economic aspect during flour drying, but reduce flour yield. The aim of this paper is to compare two methods to prepare tacca flour and to determine their characteristics. Many studies have reveal the pharmacological benefits of phytochemical compounds contained in tacca. Some researches have shown that taccalin is a cytotoxic compound found in Tacca chantrier, T. paxiana, T. plantainea, T. Subflabelata. Meanwhile the existence of taccalin in T. leontopetaloides is not mentioned explicitly (Jiang et al. 2014). Nevertheless, Borokini & Ayodele (2012) mentioned that the results of phytochemical screening on T. leontopetaloides suggests that environmental factors (difference of growing place) greatly influences the content of phytochemicals in tacca, e.g. variation content of alkaloids and cardiac lycosides. The same plant growing at different

places turn out different phytochemical content (Borokini and Ayodele 2012). In addition, another important compounds in T. leontopetaloides are flavonoids and saponins (Jiang et al. 2014). Lately, a serious concern to flavonoids have been made because of its antioxidant activity. Flavonoids are antioxidants that are widely used in the pharmaceutical world. Meanwhile the saponin compounds have the ability to affect the absorption of fats such as cholesterol and lipid metabolism in the liver (Francis et al. 2002). Test on animals found a higher lipid levels in animal feces after being given the tacca (Ndouyang et al. 2001). Thus, tacca potentially as a source of food low-fat diet or as a nutraceutical starting materials. 2. Material and Methods 2.1 Material The main material used in this study was tacca tuber from southern coast of Garut, West Java. Chemical reagen was got from local chemical ditributors. 2.2 Flouring Process Tacca was processed into flour by pressing and chipping method (Miftakhussolikhah et.al. 2014) and also be processed into tacca starch. Steps of flouring methode are: (1)pressing the grated, cleaned tuber wrapped in a cloth to dewater followed by drying,grinding and screening (Figure 1); (2) chipping the cleaned tuber followed by drying,grinding and screening (Figure 2). 2.3 Chemical Characterization The flours were characterized for proximate composition (AOAC, 1995). Carbohydrate content was estimated by mathematical calculation.

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Amylose and phenolic content was determined by spectrophotometry. Resistant starch was determined according Goni et.al. (1996) and starch content was conducted according to Nelson-Somogyi method.

specific include Dragendorf (alkaloid), LiebermannBurchard (steroids), anisaldehid (these Terpenoids), vanilin-sulfuric acid (saponin), iodine (detection of double bonds), ammonia-sitroborat (flavonoids). The spot visually observed under the rays of the visible, UV 254 and 366 (Wagner and Bladt 1996). 3. Result and Discussion 3.1 Chemical Comparison of pressed chipped tacca flour and starch

Figure 1. Flow diagram of pressing method to produce tacca flour

and

The chemical composition of pressed tacca flour, chipped tacca flour and tacca starch are presented in Table 1. Water content of pressed tacca flour (8.26%) was greater than that of made by chipping method (5.05%). Water content of tacca starch (14.44%) was greater than that of chipped tacca flour and pressed tacca flour. Water levels may affect shelf life of flour during storage and flour quality. Flour which has high water content is more easily damaged because microbes were easier to grow. Based on Indonesian National Standard (SNI) of flour, water content of cassava flour, corn starch and wheat flour should be less than 12%, 13% and 14.5% respectively (BSN, 1992). In this study, water content in pressed tacca flour, chipped tacca flour and tacca starch were 8.26%, 5.05% and 14.44% respectively. Therefore tacca flour and tacca starch meet the requirement of SNI. Table 1. Chemical composition pressed tacca flour, chipped tacca flour, and tacca starch Chemical composition

Pressed Chipped Tacca starch tacca flour tacca flour Water content (%) 8.26 5.05 14.44 Ash content (%) 0.78 1.82 0.11 Fat content (%) 0.15 0.46 0.54 Protein content (%) 1.86 5.80 0.19 Carbohydrate content (%) 89.11 87.27 84.72 Amylose content (%) 25.85 21.92 31.08 Starch content (%) 76.37 69.77 76.71 Phenolic content (mg/g) 0.3248 0.4106 0.0266 Resistant starch (mg/ml) 0.2037 0.2285 0.1036

Figure 2. Flow diagram of chipping method to produce tacca flour 2.4 Phytochemical Screening The extraction process was done by the method of multilevel maceration. The solvents used were petroleum ether, diethyl ether, and ethanol 70% respectively. The extracts derived from each solvent were then concentrated by vaporizing over waterbath. Next, extract to be spotted onto TLC plate ± 5 μl. Elution was performed with two types of eluent systems namely toluen: ethyl acetate (95:5) (system A) and ethyl acetate: methanol: water (100:13.5:10) (system B). Qualitative detection of compounds was done by spraying the reagent

Ash content of chipped tacca flour was higher than that of pressed tacca flour and tacca starch. This is due to the minerals dissolved into the water during compression process. While the ash content in Tacca starch are very low . Protein content of chipped tacca flour was higher than that of pressed tacca flour and tacca starch. This was due to tissue damage during grating and compression can cause partial discharge of water-soluble proteins. While in the starch process making, water-soluble proteins dissolve to the water. Protein content in pressed tacca flour was lower than chipped tacca flour, protein content of wheat flour in the INS is 7%.

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Amylose content of tacca starch was higher than that of pressed tacca flour and chipped tacca flour. Amylose plays a role in starch gelatinization process. Carbohydrate content of pressed tacca flour was higher than that of chipped tacca flour and tacca starch. Starch content in tacca starch was higher than that in pressed tacca flour and chipped tacca flour. Acoording to ISO 3729: 2008 about sago flour, starch content in sago flour should be more than 60%, so the starch content of tacca starch and tacca flour met the requirement. Total phenolic content of pressed tacca flour were higher than chipped tacca flour and tacca starch. The value of phenolic content of tacca starch was not different from the phenolic content of purple sweet potato varieties in range of 1.120 to 2.779 mg / 100 g(Ginting et al. 2011). Total phenolic content associated with antioxidant capacity of amaterial. Resistant starch of chipped tacca flour was higher than that of pressed tacca flour andtacca starch. According Panlasigui et.al. (1991), starch granules that rich in amylose have agreater ability to crystallize due to more intensive hydrogen bonding. Consequently, thestarch has better gelatinization at cooking time so digested more slowly. Asp & Bjorck(1992) also states that the resistant starch was higher as the higher amylose content.Resistant starch also affected positively by protein and fat content.

that there were no significant differences on the composition of phytochemicals compounds contained in the Tacca chipped-flour as well as pressed-flour as shown in Table 2. Both types of flour do not contain flavonoids that are indicated by the absence of yellow spots on the TLC post spraying KOH or NH3-sitroborate. Flavonoids in alkaline (NaOH or KOH spraying or with ammonia evaporation) will be yellow. Yellow color will be more intense with spraying of sitroborate (Mulyani and Laksana 2011). Whereas the testing of the alkaloids by spraying of dragendorf reagent it is known that these two types of flour contain alkaloids indicated by the presence of red/pink spots. Alkaloid is a secondary metabolite in the form of cyclic compound containing secondary or tertiary amine (nitrogen), so it is alkaline and tend to be soluble in polar solvents. Alkaloid has many variations ranging from simple structures (e.g. piperidin) until a complex structure (e.g. tubokurarin, poison curare in Africa) (Katavic 2005). There are currently over 27,000 compounds of alkaloids with a wide range of pharmacological ability, but less than 200 of them are commercially exploited as an antiviral, anti rheumatic, antimalarial, analgesic, antitussive, anti asthma, diuretic, aprodisiaka and so on (Amirkia and Heinrich 2014).

3.2 Phytochemical contents From the observations of thin layer chromatography by spraying the specific regaent generally known Table 2. Phytochemical contents of Tacca leontopetaloides flour

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The same also applies to the saponins, steroids, terpenoids. Three of these compounds are contained in both types of flour. The process of pressing didn't cause the significant loss of phytochemicals compounds. Steroids are compounds with steroids or sterane nucleus that is combined forming a four carbon rings (three hexagonal and one pentagonal) as shown in Figure 3. Steroid compounds are abundant in nature, spread in plants as well as animals and humans. Cholesterol is one example of steroid that is present in the human body. (Bhawani et.al. 2010).

Reference [1] Amirkia, Vafa, and Michael Heinrich, 2014, “Alkaloids as Drug Leads - A Predictive Structural and Biodiversity-Based Analysis.” Phytochemistry Letters 10: xlviii – liii. http://dx.doi.org/10.1016/j.phytol.2014.06.015. [2] AOAC, 1995, Official Methods of Analysis of AOAC International. Arlington, Virginia, USA. [3] Asp, N. G. and Björk, I., 1992, Resistant Starch, Trends Food Sci. Technol., 3: 111-114. [4] Bhawani, S A, O Sulaiman, R Hashim, and M N Mohamad, 2010, “Thin-Layer Chromatographic Analysis of Steroids : A Review.” Tropical Journal ofPharmaceutical Research 9(3): 301–13.

Figure 3. Main structure of steroids Some researcher showed that steroid is a cholesterollowering agent. Brown (2011) proposed three mechanism of how phytosterol reduces the cholesterol level, i.e phytosterol-cholesterol competition in the intestine, inhibition of protein mediated sterol transport, and interference with cholesterol-esterase enzyme. Thus, it is estimated tacca steroids also could be utilized as cholesterollowering agent in functional food. Conclusion This research revealed that there were no significant difference of water, fat and carbohydrate content in chipped and pressed Tacca flour as well as its phytochemical content. Pressing process reduces the terpenoid content but not significant. Ash, protein, amylose, and resistant starch content of chipped Tacca flour was higher than the pressed one.

[5] Borokini, T I, and A E Ayodele, 2012, “Phytochemical Screening of Tacca Leontopetaloides ( L .) Kuntze Collected from Four Geographical Locations in.” International Journal of Modern Botany 2(4): 97–102. [6] Brown, Andrew W, 2011, “Mechanisms of Decreased Cholesterol Absorption Mediated byPhytosterols in the Intestinal Lumen.” University of Nebraska. [7] BSN, 1992, Standar Nasional Indonesia: Tepung Singkong (SNI 01-2997-1992), Badan Standar Nasional Republik Indonesia, Jakarta. [8] Francis, George, Zohar Kerem, Harinder P S Makkar, and Klaus Becker, 2002, “The Biological Action of Saponins in Animal Systems: A Review.” The British journal ofnutrition 88(6): 587–605. http://www.ncbi.nlm.nih.gov/pubmed/1249308 1 (November 14, 2014).

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[9] Ginting, E., Utomo, J.S., Yulifianti, R., Jusuf., 2011. Potensi Ubijalar Ungu sebagai Pangan Fungsional. Iptek Tanaman Pangan 6(1) [10] Goni, L., L. Gracia-Diz,mas.d” and F. SauraCalixto., 1996, Analysis of Resistant Starch : Method of Food Product, J. Food Chem, 56 (4): 445-449. [11] Jiang, Jin-He, Hong-Mei Yang, Yi-Liang Wang, and Ye-Gao Chen, 2014, “Phytochemical and Pharmacological Studies of the Genus Tacca : A Review.” Tropical Journal of Pharmaceutical Research 13(4): 635–48. [12] Katavic, Peter L.. 2005, “Chemical Investigations of the Alkaloids From the Plants of the Family Elaeocarpaceae.” Faculty of Science, Griffith University. [13] Miftakhussolikhah, Dini Ariani, Cici Darsih, Mukhamad Angwar, Haryadi, 2014, Development of Flour Preparation Method Derived from Local Arrowroot (Maranta arundinacea)of Yogyakarta, Proceeding The 5th ASIAHORCs InternationalSymposium, Bali, 26-28 November 2013, ISBN: 978-979-799806-6 [14] Mulyani, Sri, and Toga Laksana, 2011, “Analisis Flavonoid Dan Tannin Dengan Metoda Mikroskopi- Mikrokimiawi.” Majalah Obat Tradisional 16(3): 109–14. [15] Ndouyang, Cj et al, 2014, “In Vivo Assessment of the Nutritional and Subchronic Toxicity of Tacca Leontopetaloides (L.) Tubers.” Scholarly Journal of Agricultural Science 4(1): 5–13. [16] Panlasigui LN, Thompson LU, Juliano BO, Perez CM, Yiu SH, Greenberg GR, 1991, Rice varieties with similar amylose content differ in starch digestibility and glycemic response in humans. Am J Clin Nutr 54:871–877. [17] Richana, N., Sunarti, T.C., 2004. Karakaterisasi Sifat Fisikokimia Tepung Umbi dan Tepung Pati dari Umbi Ganyong, Suweg, Ubikelapa, dan Gembili. Jurnal Pascapanen 1: 29-37 [18] Wagner and Bladt, 1996, Plant Drug Analysis. 2nd ed. Munich: Springer.

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Making and Characterization of Carrageenan Single Edible Film Doddy A.D, Enny S, Nok A, Novita I, Qistia H.E Researcher of Appropriate Technology Development Center, Indonesian Institute of Sciences Jl. K.S. Tubun No. 5 Subang, West java, Indonesia. Abstract: Edible film or edible coating is a hydrocolloid-based packaging material such as carrageenan. The purpose of this study is to make edible film and to get the characterization of carrageenan-basedsingle edible film. The carrageenan was used as raw materials with glycerol as plasticizer. The objective of this work was to characterize carrageenan-based edible films supplemented with glycerol by evaluating their mechanical and water vapour barrier properties. The experimental design used randomized block design with two factors, namely the carrageenan and glycerol concentration of carrageenan and the concentration of glycerol.Each factor consists of 3 levels. Carrageenan concentration level is 1%, 1.5% and 2% w/v. For glycerol concentration level is 0%, 0.5% and 1% v/v. The result show that the higher the concentration of carrageenan, the greater the tensile strength and the smaller the elongation of edible film is. Conversly, the higher the glycerol concentration, the smaller the tensile strength and the bigger the elongation of edible film is. Based on the analysis of variance, the concentration of carrageenan had no significant effect on the value of the mechanical and barrier properties. In addition, the concentration of glycerol did not significantly affect on the tensile strength and the permeability of edible film, but give the real impact tothe elongationvalue. The highest tensile strength was resulted at the carrageenan concentration of 2%. The highest elongation at break was resulted at the glycerol concentration of 1%. The lowest water vapor transmission rate was resulted on carrageenan concentrations of1%. Based on mechanical properties, the best composition ofsingle edibel film wascarrageenan:glycerol (2% w/v : 1% v/v). Keywords: edible film, carrageenan, mechanical properties. 1. Introduction Conventional packaging materials derived from petroleum and synthetic polymers that are non biodegradable negative impact on the environment has began to be abandoned (Maulina, 2012). The growing accumulation of synthetic plastic wastes together with the difficulty of recycling the majority of packaging has stimulated food and packaging industries to explore for new biodegradable packaging materials (Tharanathan, 2003). In recent years, there has been growing interest in edible films and coatings, which offer several advantages over synthetic materials, such as being biodegradable and environmentally friendly (Salmieri & Lacroix, 2006).Biodegradable edible films or edible film is a thin layer that integrates with foodstuffs, edible and can be decomposed by microorganisms. An edible coating is a thin layer of edible material formed as a coating on a food product, while an edible film is a preformed, thin layer, made of edible material, which once formed can be placed on or between food components (McHugh, 2000). Edible coating or film is defined as a continuous thin layer made of materials that can be consumed.It can be used on top or among the food products (fruits). It serves as a mass transfer barrier (water vapor, O2, CO2) or as a food additives material carriersuch as antimicrobial agents. There are three groups of base materialcoating, namely hydrocolloids (proteins and polysaccharides), lipids (fatty acids, waxes) and composite (mixture of hydrocolloids and lipids). The formed properties of coatings and films are

dependent on the characteristics of the base materials. Hydrocolloid coatings have good mechanical properties, but they have a poor barrier capacity against moisture transfer. This problem is not found in lipids due to their hydrophobic properties, especially those with high melting points such as beeswax and carnauba wax (Falguera et al., 2011). Hydrocolloids are the most widely investigated biopolymers in the field of edible coating and edible film (Falguera et al., 2011). Some of these are: starch and cellulose derivatives, pullulan, chitosan, alginate, pectin, carrageenan and konjac flour. Carrageenan is a natural carbohydrate (polysaccharide) obtained from edible red seaweeds. It has hydrophilic properties and serves as a thickener, stabilizer, coagulant, emulsifier, can form a gel, and can interact with chemicals in foods (Prajapati et al, 2014). However, the nature of edible packaging films, which is rigid and brittle, causes limitations in food applications. Therefore, to overcome film‟s brittleness and also to increase the workability and flexibility of these films, various types of plasticizers have been widely used. The primary role of a plasticizer is to enhance film fexibility and decrease brittleness. The addition of plasticizer leads to a decrease in intermolecular forces along polymer chains which improves the flexibility and alsomakes it easier for film to be peeled off from the glass plate. Glycerol being one of the most preferred and most studied (Ghasemlou et al , 2011).

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In this context, the objective of this work was to characterize carrageenan-based edible film supplemented with glycerol. This study evaluated their mechanical and water vapour barrier properties as affected by plasticizer. This study is a preliminary study to the research and characterization of composite edible film. 2. Materials and Methods 2.1 Material The research tools are glassware, petri dish, magnetic stirrer, desiccator, micrometers, Instron UTM, and SEM (Scanning Electron Microscope).The materials are carrageenan, glycerol, silica gel, tween 80, and distilled water. 2.2 Preparation of film



The single edible film based carrageenan was prepared using the method described by Tamaelaand Lewerissa (2007) that has been modified.The edibel film was prepared by dissolving carrageenan into distilled water while stirring and heated to a temperature of 70C. The concentrations of carrageenan are varied, namely: 1%, 1.5%, and 2% (w/v). Then glycerol with a concentration of 0%, 0.5% and 1% (v/v) was added to 0.2 % (w/v) Tween 80. The solution was then heated and stirred for 10 minutes at a temperature 70-80° C. Air bubbles contained in the film solution must be removed. A total of 85 mL of the film then poured on acrylic plate 20x20 cm2 subsequent drying in an oven at a temperature of 50°C for 24 hours. The film is removed and stored in a room with a temperature of 25°C for one week to be analysed. Each preparation was performed in three repetitions. 2.3 Characterization of Edibel Film Thickness Thickness of the films was measured using a handheld micrometer (Mitutoyo) by 0.001 mm precisious. Five thickness measurements were taken at random positions on each film and the mean was calculated. The mean value for the thickness was used in calculating the water vapor permeability and tensile strength.  Tensile Strength and Elongation Test Tensile strength and percentage elongation at break of the film were determined using an Instron Universal Testing Machine. Edible film was conditioned at room temperature (25 oC). Three sample strips of the film were cut from the conditioned films. The samples were mounted between the grips of the machine and test was done by clamping the end of the sampleuntil the sample broke. Value of maximum force to break the film can be seen on the the display. Tensile strengthwas calculated by dividing the maximum load by the initial cross-sectional area of the sample. Cross-

sectional area was obtained by multiplying the initial length of the sample with the initial thickness of the sample. The percentage elongation is measured based on the elongation of the film when the film broke. It was determined by dividing the extension at rupture of the film by the initial length of the film multiplied by 100. The tensile strength and percentage elongation were calculated by the following equations:

𝐹𝑚𝑎𝑥 … … … … … … … … … … … … … (1) 𝐴 𝐿 𝐸𝐵 = × 100% … … … … … … … … . (2) 𝐿0 𝜏=

where: : tensile strength (Mpa) 𝐹𝑚𝑎𝑥 : tensile force (N) A : cross-sectional area (mm2) EB : elongation at break (%) L : regastrain at break (mm) L0 : initial length (mm)  The Water Vapor Transmission Rate Test Water Vapor Transmission Rate (WVTR) was determined by gravimetrically according to the ASTM-E96-99. The petri dish with a diameter of 5 cm and containing silica gel was sealed with the film sample on the top of a dish. Then dish was put in a desiccator with a controlling condition at 60-80 % RH and temperature of 29 oC. RH inside the cell is always lower than the outside of the cell, water vapor transmission was determined by weighing the petri dish in regular time intervals for 24 h. Then be obtained slope (change in weight of the dish every time) is calculated. WVTR can be calculated using equation Shih, et al (2011) as follows:

𝑊𝑉𝑇𝑅 =

∆𝑊. ∆𝑡 … … … … … … … . . (3) 𝐴

Where: W : weight gain (g) t : time (day) A : film area (m2)

 The morphology test The morphology test was determined by using SEM (Scanning Electron Microscope).The film sample was affixed to a set holder with a double tape and then coated with gold metal in a vacuum. The sample wasplaced in SEM, then the topography image was observed by a 500 to 1000 timesof magnification. 3. Results and Discussion The carrageenan solution was prepared by dissolving carrageenan into distilled water.When carrageenan solutions were cast on a surface and dried, they leaved a film that possessed specific

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plasticity, transparent and odorless characteristics (Fig. 1). Plasticizer was added to make plasticity characteristic on edible film. To flatten the mixture, it was to add the emulsifiers as a homogenizer.

that it is more elastic (Rodirguez et al. 2006) and is relatively cheap. Howeverconcentrations of plasticizer can lead to decrease functional properties of edible films such as resistance to moisture and mechanical properties as well as the increased solubility of the film (Glicksman, 1984). Tween 80 was used as the edible film emulsifier. This material has polar and non-polar groups so it can be a liaison between the hydrocolloids (carrageenan). The use of emulsifiers can alter the surface tension between the coating and the coated material on edibel coating (Debeaufort and Voilley 2009). 3.1 Characteristics Edibel Film

Figure1. Carrageenan-based edible film Plasticizers are additives that are used to change the flexibility and the mechanical properties of the film from the film matrix (Dangaran et al. 2009). Plasticizer used in this study is glycerol, because it has hydroxyl groups that can form hydrogen bonds that can degrade the polymer intermolecular force so

0 0.5 1

0.030 0.039 0.045

Carrageenan

Single

The objective of the edible film production is to get packaging material that has mechanical properties and barrier properties to the product environment. Thecharacterization (mechanical and barrier properties) of single edible film on various carrageenan and glycerolconcentrations are presented in Table 1.

Table 1. Characteristics of a single edible film of carrageenan Carrageenan Glycerol Thickness Tensile concentration concentration (mm) strength (% b/v) (v/v) (MPa) 0.027 1 0 21.80a 0.031 1 0.5 21.96a 0.031 1 1 26.59a 0.023 1.5 0 29.91a 0.031 1.5 0.5 42.98a 0.032 1.5 1 41.61a 2 2 2

of

32.95a 44.32a 42.60a

4.5a 8.4a 19.4b

Water vapor transmission rate (g.m-2.24 hour -1) 48.60a 49.96a 50.46a

2.8a 7.5a 18.4b

52.86a 50.15a 52.97a

3.5a 4.1a 13.3b

51.19a 49.18a 51.39a

Elongation (%)

* Different letters indicate the test results were significantly different (P <0.05)

 The thickness The thickness of the edible films is very important for the physical properties of the films. The average thickness values for all carrageenan-based films were from 0.027 to 0.045 mm.The greater carrageenan and glycerol concentration, the higher the film thickness is.  Mechanical properties Tensile stress is defined as the maximum voltage to be able to withstand the pull of the film. The elongation at break indicates the ability the length of edible film. Table 1 shows the tensile stress and elongation at break for the carrgrenan-based edible films. The higher the tensile strength and the elongation, the better the quality of the film is. As a

result the film is not easily broken when used to package the product. The greater the concentration of carrageenan, the higher tensile strength value and the lower the elongation value is.The highest tensile strength value of carrageenan edible film is at the carrageenan concentration of 2% with a value above 32 MPa. The value of this tensile strength is in accordance with standards that can be applied to edibel the film, which is 10-100 MPa (Krochta and Johnston, 1997). The greater the concentration of carrageenan, the bigger the polymer matrix will be and the better the intramolecular interaction is. The concentration of glycerol for the best tensile strength value and elongation at break is 1% (v/v) in

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the carrageenan film edibel. This value is consistent with the statement of Krochta and Johnston (1997) that the characteristic of standard edibel film has a value of 10-50% elongation.Based on Duncan test tensile strength values of carrageenan edible film is not significantly different (P> 0.05) for each treatment. Meanwhile, the elongation values were significantly different (P <0.05) at glycerol concentration of 1% (v / v). Tensile strength of the films moderately decreased as the glycerol concentration increased, converselythe elongation break increased (Table 1). Glycerol was added to decrease the matrix stiffness and to increase mobility so that it is more flexible(Yoshida, 2009).Thus, addition of glycerol to carrageenan film increased its extensibility, but reduced the mechanical strength. Polar groups (-OH) along plasticizer chains are believed to develop polymer-plasticizer hydrogen bonds replacing the polymer-polymer interactions in the biopolymer flms. Incorporation of glycerol molecules into carrageenan films probably established glycerolcarrageenan hydrogen bonds replacing some of the carrageenan-carrageenan hydrogen bonds. As a result, direct interactions between the molecular chains were reduced, and the chain segmental mobility was increased, causing the mechanical strengthof the flms to be decreased and the extensibility enhanced (Yang, 2000).  The barrier properties Edibel film barrier properties consist of water vapor permeability, gas permeability (O2 and CO2), volatile permeability and permeability of the solute. The tested barrier properties were permeability to water vapor because of the hydrophilic nature of the polysaccharide-based films have a limited inhibition of the ability of water vapor barrier (Lacroix, 2009). Tabel 2 shows the water vapor transmission rate value at each film.Based on Duncan test, the values of WVTR are not significantly different (P> 0.05) for each treatment. The value of WVTR is high because the ediblefilm hydrophilic nature of the material. One of themajor purposes of edible films is to block moisture transferbetween the food and the surrounding atmosphere, therefore theWVP should be as low as possible. Table 1shows that the film WVTR increased with increasing glycerol in the fillm.Generally, water vapour transmission through ahydrophilic fillm depends on both difusivity and solubilityof water molecules in the film matrix. An increase in the interchain spacing due toinclusion of glycerol molecules between the polymerchains may promote water vapour difusivity throughthe film and hence accelerate the water vapour transmission. The high hydrophilicity of glycerol moleculeswhich is favourable to the adsorption of water

moleculescould also contribute to the increase in the film WVTR (Yang, 2000). The best composition of carrageenan single film is: glycerol (2% w/v: 1% v/v) because it has the best elongation and tensile strength. This is in accordance with Tamela and Lewerisa (2007) that the best composition of carrageenanedibel film is (2% w/v: 1% w/v).  The morphology test Scanning electron microscopy (SEM) image of the carrageenan-based edible film is shown in Figure 2.

Figure 2.Surface morphological of carrageenanedible film From this image shows that the edibel film surface was not smooth, heterogeneous and porous. The heterogeneous surface was due to air bubbles formed among its constituent materials (Wicaksono, 2012). The bubble was formed as a result of temperature instability resulting upper surface of the ediblefilm getting dried faster so it would evaporate water molecules trapped among layers inediblefilm. Debeaufort and Volly (2009) stated that the solvent evaporation would be faster at higher temperatures and the film will dry faster. Heterogeneous film surface could be correlated with a high water vapor film transmission rate. Water vapor transmission rate of single edible film was larger than the composite as a single film surface is more heterogeneous than composites. This is consistent with Yoshida (2009) that the heterogeneity of the matrix can form an empty space or cracks that may affect permeability. 4. Conclusions and Recommendations 4.1 Conclusion Single edibel film can be made from local raw materials which are abundant in Indonesia;it is carrageenan (from seaweed).The higher the concentration of carrageenan, the bigger the tensile strength and the smaller the elongation of edible film is.Conversly, the higher the glycerol concentration, the smaller the tensile strength and the bigger the elongation of edible film is. The highest tensile strength is resulted at the carrageenan concentration

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of 2%. The highest elongation at break is resulted at the glycerol concentration of 1%. As the nature of water vapor permeability, the lowest value on carrageenan concentrations is at 1%. Based on the analysis of variance, the concentration of carrageenan had no significant effect on the value of the mechanical and barrier properties. The concentration of glycerol did not significantly affect the tensile strength and the permeability of edible film, but give the real impact tothe elongationvalue. Based on the elongation value and the tensil strength, the best carrageenan single ediblefilm has the composition of carrageenan : glycerol (2% w/v: 1% v/v). This film has the thickness of 0.045 mm, the tensile strength of 42.60 MPa, the elongation at break of 13.3%, and the water vapour transmission rate of 51.40 g/m2/24 hour. 4.2 Recommendations The outcome of edible film was not homogeneous so that itneedsfurther study in making edible filmcomposite by emulsifier. The mixing process of the edible film is better used homogenizer in order to get homogeneous film solution. In addition further tests need to be done to analyze the structure and thermal tests on edibel films.

[6]

KrochtaJM and Johnston CDM. 1997. Edible and biodegradable polymer films:challenges and opportunities. Food Technology 51(2):6174.

[7]

Lacroix M. 2009. Mechanical and permeability properties of edible film and coatings forfood and pharmaceutical applications. In: Embuscado ME. Huber CH, editors. EdibleFilms and Coatings for Food Applications. USA (US): Springer. pp 347366. Maulina J. 2012. A study of edible film from pulp cacao. Majalah Polimer Indonesia 15 (1):28–36 McHugh T H. 2000. Protein-lipid interactions in edible films and coatings. Nahrung 44: 148-151. Prajapati, Pankaj M. Maheriya, Girish K. Jani, Himanshu K. Solanki. 2014. Carrageenan: A natural seaweed polysaccharide and its applications.Carbohydrate Polymers 105: 97– 112 Rodriguez M. Oses J. Ziani K.and Mate JI. 2006. Combined effect of plasticizers and surfactants on the physical properties of starch based edible films. Food Research International 39:840-846. Salmieri S. and Lacroix M. 2006. Physicochemical properties of alginate/polycaprolactone-based films containing essential oils. Journal of Agricultural and Food Chemistry 54: 10205– 10214. Shih FF. Daigle KW. Champagne ET. 2011. Effect of rice wax on water vapour permeability and sorption properties of edible pullulan films. Food Chemistry 127 (2011): 118–121 Tamaela P.And Lewerissa S. 2007.Karakteristikedible filmdari karaginan.Ichthyos 7(1):27-30. Tharanathan R N. 2003. Biodegradable films and composite coatings: Past, present and future. Trends in Food Science and Technology 14: 71–78. Yang Y. Paulson AT. 2000. Mechanical and water vapour barrier properties of edible gellan films. Food Research International 33: 563-570 Yoshida CMP. Junior ENO. Franco TT. 2009. Chitosan tailor-made films: the effect of additives on barrier and mechanical properties. PackagingTechnology Science 22:161-170

[8]

[9]

[10]

[11]

Acknowledgements The authors would like to express depp thanks to Materials Development and Manufacturing Engineering Research Program in 2015 and Appropriate Technology Development Center – LIPI for kindly sponsored this works.

[12]

References [1]

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[3]

[4]

[5]

[ASTM] American Society for Testing Material. 1999. Annual Book of ASTM Standards. Philadelpia (US). Dangaran K. Tomasula PM. Qi P. 2009. Structure and function of protein-based edible film and coatings. In: Embuscado ME. Huber CH, editors. Edible Films and Coatings for Food Applications. USA (US): Springer. pp 25-56. Debeaufort F and Voilley A. 2009.Lipidbased edible films and coatings. In: Embuscado ME. Huber CH,editors. Edible Films and Coatings for Food Applications. USA (US): Springer. pp 135-168. Falguera V. Quintero JP. Jimenez A. MunozJA. and Ibarz A. 2011. Edible films and coatings: Structures, active functions and trends in their use. Trends in Food Science &Technology 22: 292-303. Ghasemlou M. Khodaiyan F. and Oromiehie A. 2011. Physical, mechanical, barrier and thermal properties of polyol-plasticized biodegradable edible film made from kefiran. Carbohydrate Polymers 84: 477–483.

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The Development of Traditional Food-Based Military Ration Packed in Cans M. Kurniadi, A. Nurhikmat, M. Angwar, A. Susanto, Tri Wiyono, A. S. Praharasti UPT.Balai Pengembangan Proses danTeknologi Kimia-LIPI Yogyakarta Desa Gading,Kec. Playen,Gunungkidul,Yogyakarta Phone. (+62) 274-392570, Email: [email protected] Abstract: Military ration is one kind of foods that has been prescribed dose of nutrition according to their needs. Food for military personnel should remain stable, durable, safe, practical and does not ignore the comfort aspect. The aim of this study is to make the traditional foods-based military rations packaged in cans and to find out its sterilizing value (Fo). The study was conducted in two stages. The first stage is to formulate the traditional foods and package them in cans. The traditional foods’ formulations contain of 4 formulas of nasi uduk and 5 formulas of fried rice. The type of can which is used is the Pop-end models with the size of 301x205. The second stage is the process of canning the food, sterilization Fo value measurement, the process of canning, chemical testing, microbiological and sensory testing. The data of the testing result is processed statistically and descriptively. The results shows that the ration in cans with the highest energy content is formula NG-3 (fried rice + chicken + beef sausage) with 479.128 Calories, and formula NG-1 (rice fried + egg + chicken) with 433.894 Calories, the value of Fo sterilization from the NG-3 formula and NG-1 respectively 11.97 and 11.11 minutes. The organoleptic test shows that panelists prefer to NG-1 than NG-3. Keywords: Military rations, traditional foods, cans, Fo sterilization value 1. Introduction In the development of the armed forces, one of the aspects that influence the success of a military operation is the logistics supply to army personnel. (Ditbekangad, 2011) One of them is the food nutrition. Food supply for military personnel should remain stable/durable, safe, practical and have the comfort aspect when they are consumed. Amount Dietary Allowance (ADA) of Indonesian soldiers which is reported by Setyowati (2008) states that as long as troops are in the barracks, the soldiers' needs of nutrition and energy have been met from the organization of foods in the hostel kitchen for 2772 Cal/day plus 219 Cal/day from the cafeteria that meets 74.3% - 88.6% ADA. One of the military occupational healths is the soldiers' nutrition in the field that covers the nutritional needs of soldiers in training, nutrition soldiers in combat. To achieve the optimal health status, some nutrition is absolutely necessary needed and they can be obtained from foods in accordance with the recommended amount in each day. The souldiers‟ energy sufficiency, Tharion et.al (2005) are ranging from 3109 Cal to 7131 Cal per day. To overcome the saturation of the main food menu then the US military has been utilized Pizza as the distinctive food to be used as the supplementary food (snack) for the troops on the battlefield. The food is made to last for 3 years (Gizmag, 2014). Indonesia has a lot of traditional food that deserves to be developed as foods for soldiers in the battlefield such as yellow rice, nasi uduk, nasi gudeg and others.

Foodstuffs can be preserved by heat sterilization. However, every food has its specific heat transfer, which is influencing the effectivity of sterilization. The critical factors which should be considered as it is can affect the thermal process namely: (a) material of the product (pH, consistency/ viscosity of the material, the shape/ size of materials, water activity, percent solids, the ratio of solids/ liquids, changes in the formula, the particle size, type of thickener, type of preservative added, etc.), packaging (type and dimensions, the method of filling into the packaging), (b) the retort (types of retorts, media type heaters, the position of the container in the retort, stacks of container, setting the cans, the possibility of nesting) (Kusnandar et. al, 2013). Thermal processes such as sterilization involve the transfer of heat by conduction and convection. By conduction, heat transfer occurs directly from the particles without passing through any other medium other than the product itself. Moreover, in the convection, the heat transfer occurs through the liquid medium surrounding the fish. The effectivity of sterilizaton is estimated by the F value which is calculated from the temperature of the process and the Z value of microbes. The Fo value is the time (in minutes) at 250 ° F is required to destroy a certain number of microbes that have the Z value which is equal to 18 ° F. Resistance or resilience of cells and spores of microorganisms to heat different among microorganisms. In general, microorganisms are more resistant to heating at neutral or near-neutral pH. Thermal Death Time (TDT) at temperatures of 121 ° C has been used as reference sterilization and expressed as Fo.

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To Clostridium botulinum Fo its value from 2.45 to 2.8 minutes (Urbain, 1971 in Soeparno (2005). To determine the TDT or Fo, it is used the equations presented by Lewis (1987), {(T 121 / 10 )}

log L 

T  121 or L  10 10

…................... (1)

Yogyakarta. Empty cans with the size of 301 x 205 made by PT.Cometa Can Tangerang, chemicals for chemical analysis and test rancidity proximate analysis (TBA), microbiology material for microbiological analysis TPC. 2.2 Tools

The aim of this research is to make the traditional foods-based ration in the cans and to find out the strearilization value (Fo) of the canning process.

Tools used include canning line system such as an electric autoclave, exhauster, and tin cover automatic machines. Chemical and microbiological analysis tools such as analytical balance, tools laboratory glassware, Kyeldahl analyzer, soxhlet, Fo-meter (Ellab), oven, pH meter, incubator, petri dishes and a set of sensory test equipment.

2. Methodology

2.3 Foods formulation

2.1 Materials

The formulas of 4 nasi uduk and 5 fried rice are made by composition rice: side dish (70:30). Net weight of cans of size 300 x 205 is 160 grams so the composition is 112 grams of rice and side dishes 48 grams. The detail formulation of nasi uduk and fried rice are presented in Table 1 and Table 2.

Where Fo can be measured with the equation of:

Fo   Ldt …………….................................. (2)

Materials used in this study are rice, fried chicken, chicken eggs, beef sausage, shredded beef, onion, garlic, butter, cooking oil, salt, coconut milk obtained from Wonosari Market, GunungKidul Table 1. Formulation of the nasi uduk

Table 2. Formulation of fried rice

2.4 The canning process The canning process includes the following stages: cooking, exhausting for 5 minutes, automatic sealing, sterilization at 121 ° C for 15 minutes using alectric autoclave, cooling by water, and quarantine for 14 days (Winarno, 1994). 2.5 The determination of the Fo Sterilization Value Fo value measurement. The aim is to determine the value of the sterilization process of canning. It is conducted in the stage of the sterilization process at a temperature of 121oC for 15 minutes, using the

Ellab Fo-meter, 4-channel, metal sensor with a diameter of 1 mm and an accuracy of 0.01. The data is processed and analized mathematically. 2.6 The Measurement Parameter Testing parameters measured were physical, chemical, microbiological and sensory testing. Physical test is conducted by observing the visually odor, color, taste, aroma, texture. The test chemicals includes the proximate analysis of the water content, ash content, fat content, protein content, and carbohydrate content (AOAC, 1995), the degree of rancidity with TBA Method (Apriyantono, 1989),

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and pH. The microbiological test is the determination of TPC (SNI 01-2891-1992) (BSNRI, 1992). The organoleptic test is conducted by the scoring method (Kartika et.al, 1990). 2.7 Data Processing The data of physical, chemical and microbiological testing were analyzed descriptively. Fo the sterilization value, the measurement of data was processed by the method of the graph. Sensory test data were graphically analyzed. 3. Result and Discussion

rice in cans that have been saved for 6 months, it shows that the quality is still good and normal, but the texture is a bit hardening. 3.2 The proximate analysis The military's main food must contain nutritional value and high energy. The nutrient content result shows that the protein content is generally quite high (3.46 to 7.49%), with the fried rice formula of NG-1 (fried rice + chicken + chicken eggs) at least it is as high as 7.40%, the carbohydrate content of fried rice is average higher than nasi uduk. The proximate analysis result is presented in Table 3.

3.1 The physical testing The result of the visual observation on the color, flavor, smell and after-taste to nasi uduk and fried Table 3. Results of the proximate analysis of nasi uduk and fried rice in cans

For comparison, it is also conducted the testing of the proximate to the military rations in cans (T2SP), plastic packaging (T2P), pouch packaging (T2PJ)

and aluminum foil packaging (FD3). TNI ration proximate analysis results are presented in Table 4 below.

Table 4. Results of the proximate analysis of the TNI Rations

3.3 The composition of energy The average composition of energy in a canned fried rice is greater than canned nasi uduk. With the can sized of 301 x 205, the food weight net is 160 grams. The energy composition of the total of fried rice with the formula of NG-3 (fried rice + chicken +

beef sausage) is 479.128 calories, followed by the next formula of NG-1 (rice fried + egg + chicken) is 433.894 calories. The need of the energy of the TNI combat forces per day is 3000 Cal, so for a day need is ± 7 cans of 160 grams each.

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3.4 The test of rancidity (TBA Numbers) To determine whether the quality of canned foods is still good or is damaged, one of which is to test the rancidity (TBA Numbers). Rancidity test results on nasi uduk and fried rice packed in cans that has been stored for 1 month showed the absence of signs of

damage due to the occurrence of rancidity. The numbers of TBA which was detected between 0.0487 to 0.1533 is still far from the threshold of rancidity as it is 0.9000. Rancidity test result is presented in Table 5.

Table 5. The result of the micobiology, pH and rancidity (TBA Numbers) test

3.5 The Test of Microbiology (TPC numbers) Besides applying the chemical means, to determine whether the quality of canned food is still good or is damaged, one of which is by the microbiological testing (TPC numbers) and identifying the presence of the C. botulinum pathogenic bacteria. The result of the microbiological tests on nasi uduk and fried rice packed in can that has been stored for 1 month showd the absence of any signs of deterioration due to thr microorganisms. The TPC numbers show between 1.9 x 102 s / d 8.5 x 103. The formula of NG-3- is the lowest and NU4 is as the highest. C.botulinum pathogenic bacteria was not detected, it

means that the sterilization runs with sufficient heat as needed. 3.6 The Sensory Testing The organoleptic test results on two types of fried rice processed packed in the can is presented in Figure 1 below. Fifty one point six percent (51,6%) of the panelist prefer to NG-1 formula while the rest of them prefer to NG-3. The preferency values ranged from 3 (netral), 4 (like) and 5 (really like), resulting average value of 3,68 (NG-1) and 3,35 (NG-3).

Figure 1.Panelists preferency value of organoleptic test

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3.7 Fo Test (Sterilization Value) Fo measurement is important to know the efficiency value of commercial sterilization at 121°C for 15 minutes. Not all food commodities have similar condition. Fo value measurement results for nasi uduk and fried rice are different. Nasi uduk has a greater Fo value than the fried rice, means that for a sterilization of nasi uduk rice which contains coconut milk requires greater heat than fried rice which has a lower water level. The measurement with Fo meter produces the following Fo chart, Figure 2-4.

contains a lot of fat has a lower specific heat than the one contains less fat, so that for the increase of the temperature requires less heat energy (Forrest et al, 1975). 4. Conclusion According to the results of the research and discussion above, it can be concluded that: (1) Military rations in cans with the highest energy content are NG-3 formula (479.128 Cal), and NG-1 formula (433.894 Cal), (2) The sterilization value of Fo of NG-3 and NG-1 formula are 11.97 and 11.11 minutes respectively, (3) The organoleptic test results show that panelists prefer to NG-1 than NG3. Acknowledgement This research was funded by Program Riset Unggulan LIPI 2015. Reference [1].

AOAC, 1995, Official Method of Analysis, 14 th. Edition. Published by The AOAC Inc.Virginia. USA

[2].

Apriyantono, A. et.al.,1989, Analisis Pangan, Pusbangtepa IPB Bogor. BSN-RI, 1992, Standar Nasional Indonesia: Cara Uji Makanan dan Minuman, Jakarta: BSNRI.

[3].

Ditbekangad, 2011, Perbekalan Militer, [Online] http://www.ditbekangad.mil.id/ index.php?option=com_content&view=article &id=72&Itemid=63/ accessed on 29 Nopember 2011.

[4].

Forest, J.C., Aberle, E.D., Hendrick, H.B., and Merkel, R.A., 1975, Principles of Meat Sciences, W.H. Freeman and Co, San Fransisco

[5].

Gizmag, 2014, Pizza Khusus Untuk Pasukan Militer Awet Sampai 3 Tahun. [Online] http://lifestyle.liputan6.com/read/2051586/piz za-khusus-untuk-pasukan-militerawetsampai3-tahun#sthash.gX7F8qyE.dpuf. Accessed on 19 May 2014.

[6].

Kartika, Bambang, 1990, Pedoman Uji Inderawi Bahan Pangan, PAU Pangan dan Gizi. Yogyakarta.

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Kusnandar, F., Hariyadi, P., dan Wulandari, 2013, Parameter Kecukupan Proses Termal. Teknologi Pangan UNIMUS.

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Lewis, M.J., 1987, Physical Properties of Foods and Food Processing System, Ellis Horwoods Ltd, Chichester, England.

[9].

Setyowati, Ratih Dewi, 2008, Sistem Penyelenggaraan Makanan, Tingkat Konsumsi, Status Gizi Serta Ketahanan Fisik

Figure 2. Nasi Uduk + Chicken Egg + Beef Floss (Fo 31,77 Min)

Figure 3. Fo Fried Rice + Chicken Egg + Chicken Meat, NG-1, (11,11 Min)

Figure 4. Fo Fried Rice + Chicken Egg + Beef Sausage, NG-3, (11,97 Min). From Figure 3 and 4 above show that Fo for formula NG-1 (Fried Rice + Chicken Egg + Chicken Meat) is 11.11 minutes and for NG-3 (Fried Rice + Chicken Egg + Beef Sausage) is 11.97 minutes. The rate and magnitude of the temperature increase for fried rice depend on the temperature differences between the heat source with the processed fried rice and the duration of use of the heat. A meat which

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Siswa Pusat Pendidikan Zeni Kodiklat TNI AD Bogor Jawa Barat. Fak. Pertanian, IPB. [10]. Soeparno, 1992, Ilmu dan Teknologi Daging, Gadjah Mada University Press, Yogyakarta. [11]. Tharion, William J et al., 2005, “Energy Requirements of Military Personnel.” Appetite 44(1):47–65. http://www.sciencedirect.com/science/article/ pii/ S019566630400128X (December 24, 2014). [12]. Winarno, F.G., 1994, Sterilisasi Komersial Produk Pangan, Gramedia Pustaka Utama, Jakarta.

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Screening of α-glucosidase Inhibitor-Producing Lactic Acid Bacteria from Ganoderma lucidum as Functional Food Candidate for Diabetic Rifa Nurhayati1), Andri Frediansyah1), Fitriana Rahmawati2), Endah Retnaningrum2) and Langkah Sembiring2) 1)

Technical Implementation unit for Development of Chemical Engineering Processes 2) Faculty of Biology, Gadjah Mada University Email: [email protected]; [email protected]

Abstract: Ganoderma lucidum has widely known as herbal medicine for diabetes. Recently, alphaglucosidase inhibitors (AGI) have also been reported for their ability to reduce blood sugar level in type 2 diabetes. The enzyme inhibitors can be derived from bacteria that live in Ganoderma lucidum as their host. The aims of this study were to isolate and conduct AGI-producing endophytic lactic acid bacteria screening from Ganoderma lucidum. Fourteen isolates were identified as lactic acid bacteria. The crude extracts were examined to measure their alphaglucosidase inhibition activity. The results showed that the crude extract from L17 isolate had the highest AGI activity out of 14 isolates. According to Line-Weaverberg plot, L17 crude extract was competitive inhibitor and identified as Lactococcus sp. In conclusion, L17 isolate from Ganoderma lucidum has potential to produce AGI that could be used as functional food alternative for diabetes. Keywords: alpha-glucosidase inhibitor, Ganoderma lucidum, lactic acid bacteria 1. Introduction Ganoderma lucidum or commonly called lingzhi, a white rod basidiomycetes, is well known as antitumor through immunomodulation and antiangiogenesis (Bob et al., 2007; Zengtao Xu et al., 2011). It also has various component which have been recognized as antidiabetic therapy (Pan et al., 2013; Ma et al., 2015) such as polysaccharides, triterpenoids, glycoprotein and protein (Ma et.al., 2015; Fatmawati et.al., 2009). Aqueous extract of Ganoderma lucidum decreased the blood sugar level of alloxan diabetic Wistar rats (Mohammed et al., 2007). Various strategy have been applied for type 2 diabetes therapy, such as stimulation of insulin secretion, enhancement of insulin activity in target tissue, oral consumption of hypoglycemic agent such as biguanids and sulfonilurea, as well as inhibition of starch degradation by α-glucosidase. Α glucosidase converts oligosaccharides and disaccharides to glucose located in the small intestine epithellium (Sudha et al., 2011). Inhibition of α-glucosidase is able to slow down carbohydrate metabolism and absorption, thereby lowering blood glucose levels, a common strategy to diabetic theraphy. According to Loranza (2012), current synthetic α-glucosidase inhibitors clinically used are Acarbose and mignitol but it has limited ability, non specific, with side effects such as indigestion, bloating, nausea, abdominal discomfort and diarrhea. Beside chemical synthesis, Acarbose can also be obtained from aerobic gram-positive bacteria, member of genus Actinoplanes (Wehmeier & Piepersberg, 2004).

Strobel & Daisy (2003) mentioned several bacteria living associated with plant or fungi are able to produce bioactive compounds similar to the hosts. To intensify bioactive compounds production without directly extracting from the host, its indigenous microbe can be isolated and utilized (Simarmata et al., 2007). Research on bioactive compound-producing bacteria similar to the hosts could be an alternative solution to overcome drugs dependence and prevent medicinal plants excessive exploitation. Recently, there are several researches on lactic acid bacteria (LAB) producing αglucosidase inhibitor. One of them reported that LAB filtrate hydrolyzing skim milk was able to inactivate α-glucosidase (Ramchandran, 2008). However, research on αglucosidase inhibitor LAB isolated from Ganoderma lucidum has not been reported. Therefore, a research on inhibitor-producing LAB from Ganoderma lucidum was needed to obtain alternative, particularly to prevent drug dependence T2DM treatment as well as to prevent medicinal plants excessive exploitation. This present research aimed to isolate LAB from Ganoderma lucidum and to measure their ability to produce α-glucosidase inhibitor secondary metabolite. 2. Methods 2.1 Isolation and purification of lactic acid bacteria Source of isolate was 4 days – ripened lingzhi. Sample was washed and its surface was sterilized using Coobs and Franco method (2003) by soaking in 70% alcohol solution for 1 minute, then in 1% natrium hypochlorite for 5 minutes, and in 70% alcohol again for 1 minute before finally rinsed in

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sterile distilled water. LAB was isolated using Adebayo & Onilude method (2008) with slight modification. Approximately 10 g of sample was mashed and put into 90 ml 0.85% NaCl solution, and diluted in serial dilution up to 10-3. For each dilution, 1 ml was put into sterile petridish, added with 1520 ml MRS medium and 1% CaCO3. Inoculum was then incubated at 37°C for 48-72 hours. Suspected LAB formed clear zone around colonies. Different colonies with various color and size were then streaked across a quadrant on MRS agar medium, incubated at 37°C for 72 hours. Streaking was repeated to yield a pure culture. Isolates were selected for LAB by key characteristics identification, such as negative result for catalase test, gram-positive, non-motile and nonspore-forming. 2.2 Culture growth of α-glucosidase inhibitorproducing LAB isolate Bacteria isolate was grown according to Chen et al. (2004) in broth medium consisted of 0.1% soluble starch, 0.5% peptone and 0.15% yeast extract at pH 7. Culture was incubated for 5 days with 120 rpm agitation in room temperature. Cells were separated using centrifugation at 4000 rpm for 20 minutes, then supernatant was taken for α-glucosidase inhibition activity measurement. 2.3 Measurement of α-glucosidase inhibition activity of selected isolate Inhibition activity against α-glucosidase was measured using combined method (Bachhawat et al., 2011; Shaheen et al., 2006) with slight modification, based on substrate breakdown by the enzyme into stained product whose absorbance was measured during certain time. α-glucosidase (SIGMA) was diluted into 0.1 M phosphate buffer pH 7 at 0.75 unit/ml. Substrate was 20 mM p-nitrophenyl-alphaD-glucopyranoside (Calbiochem) diluted into similar buffer solution. Mixture consisted of 35 µl of substrate, 68.6 µl of 0.1 M phosphate buffer pH 7 and 4.2 µl of sample was put into microplate (Brand Plates). After allowed to react in 37°C incubation for 5 minutes, 35 µl of enzyme solution was subsequently added and incubated for 15 minutes at 37°C. Reaction was stopped using 70µl of 200 mM Na2CO3 solution. Absorbance of nitrophenol produced then measured using Elisa reader (Thermo Scientific) at λ 400 nm, with 1% acarbose solution (Fluka) as reference. Inhibition activity against αglucosidase was measured using following equation: % Inhibition =

(Ab - Ac) - (As - Ac) x100 % (Ab - Ac)

Where: Ab = blank absorbance; As = sample absorbance; Ac = control absorbance (Pujiyanto et al., 2012)

2.4 Measurement of α-glucosidase inhibition activity Kinetic inhibition of selected isolate with highest activity was then measured, with and without sample presence at various substrate concentrations of 1mM, 2 mM, 5 mM, 10 mM and 15 mM. Measurement was conducted by Lineweaver-Burk plot analysis to obtain MichaelisMenten kinetics constant (Elya et al., 2012), calculated based on nonlinear regression equation y= a+bx, with 1/[S] as xaxis and 1/v as y-axis (Murray et al., 2009). 2.5 Identification of α-glucosidase inhibitorproducingLAB isolate Selected isolate with inhibition activity αglucosidase was further characterized based on Putri et al. (2012) using commercial kit API 50 CHL (Biomerieux, France). Twenty four hours old isolate was streaked on MRS medium to obtain considerable amount of pure culture before suspended into API 50 CHL medium and homogenized. Isolate suspension in the medium then dropped on API 50 CHL strips containing 49 kinds of carbohydrate as substrate, which then incubated at 37 °C for 48 hours. Isolate ability to ferment substrate was observed after 24 and 48 hours. Color changing from dark blue to yellow was considered as positive result. 3. Results and Discussion 3.1 Isolation and purification of acid-producing bacteria There were 20 colonies of acid producing bacteria obtained, indicated by clear zone around each colony on MRS + 1% CaCO3 medium. Isolates obtained from Ganoderma lucidum had similar morphology of milky white circular with predominantly coccus cellular shape. Acidproducing bacteria then underwent initial screening for LAB identification based on 4 key characteristics mentioned in Bergey‟s Manual Determination of Bacteriology: gram staining, catalase test, endospore staining and motility (Holt et al., 1999). Among 20 isolates, there were 14 isolates identified as lactic acid bacteria, indicated by catalase-negative, grampositive, negative endospore and negative motility. Screening results are presented in Table 1. According to Bergey‟s Manual Determination of Bacteriology, 14 isolates obtained from Ganoderma lucidum were categorized as LAB. Isolation of LAB from various organism sources, plants or animals, have been reported by various researches. This present study was able to obtain 14 LAB isolates from Ganoderma lucidum, indicated that Ganoderma lucidum was also LAB habitat. To date, isolation of lactic acid bacteria from its Basidiomycete has never been done before.

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Table 1. LAB isolate screening results

3.2 Measurement of α-glucosidase inhibition activity of LAB isolate Measurement of α-glucosidase inhibition activity was conducted on 14 LAB isolates previously obtained from Ganoderma lucidum to determine inhibition percentage of active compounds of each isolate. Results showed that 14 tested isolates were able to inhibit αglucosidase activity. Inhibition percentage values against α-glucosidase are presented in Table 2. Based on data in Table 2, L17 was the highest inhibition activity (6.28%), while acarbose activity as reference was 11.04%. The inhibition activity from L17 was lower than acarbose because the extract (supernatant) was not pure compare acarbose. Beside that, the intracellular extract was not measured. Chen et al. (2014) mentioned that αglucosidase inhibitors produced by LAB were both intracellular and extracellular.

Research on α-glucosidase inhibition producing bacteria is plenty. Pujiyanto & Ferniah (2010) obtained PR-3, endophytic bacteria from bitter gourd proven to inhibit α-glucosidase activity. Another research was following in 2012, when Pujiyanto and others were able to obtain 5 Actinomycetes isolate from various medicinal plants often used for diabetes treatment. These all indicated that bacteria living and associate with the host have potential to produce similar metabolites due to mutualism relation. To date, various researches have been directed on the quest of LAB new strains with ability to produce α-glucosidase inhibitor as probiotic. Ramchandran (2009) studied the ability of L. casei and L. acidophilus to inhibit α-glucosidase and their potential as probiotic. Similar research was conducted by Chen et al. (2013) selected 11 LAB strains which had probiotic potential as well as able to inhibit α-glucosidase activity.

In human‟s digestion system, carbohydrate is metabolized into simple sugar, monosaccharide and disaccharide, from oral consumption to small intestine. Abnormality occurs in hyperglycemia patient, particularly inside the body of those living with T2DM, where glucose level is high due to cellular inability to absorb it. This condition triggers disruption on various metabolism reactions and detrimental to the organs function (Guyton & Hall, 2007). Since carbohydrate breakdown into glucose in small intestine is catalyzed by α-glucosidase, enzyme inhibition is expected to reduce glucose accumulation in circular blood. Inhibitor compounds produced by LAB isolate is expected to be an alternative treatment of hyperglycemia and T2DM in particular. 1st International Conference on Appropriate Technology Development 2015, Bandung, October 5th-7th, 2015 | 78

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Table 2. Percentage of α-glucosidase inhibition by LAB isolated from Ganoderma lucidum

3.3 Kinetic measurement inhibition activity

of

α-glucosidase

The kinetic was measured to determine inhibition type of bioactive compounds in L17 against αglucosidase, whether competitive or non-competitive inhibition. Inhibition type was determined based on data analysis through Lineweaver-Burk plot to calculate Michaelis-Menten kinetics constant (Elya et al., 2012) using regression equation of y = a + bx, with 1/[S] as x-axis and 1/A as y-axis (Murray et al., 2009). Based on Lineweaver-Burk plot (Figure 1.), when 1/[S] close to 0, maximum reaction velocity (Vmax) was not affected by inhibitor. Therefore, at high substrate concentration, Vmax of system with inhibitor was similar or close to Vmax without inhibitor. The results indicated the inhibition was competitive since Vmax was not affected, Km values increased instead (Murray et al., 2009) Calculation on Michaelis-Menten kinetic constant resulted constant Vmax for both system, with and without inhibitor, and Km was higher on inhibitor system compare to non-inhibitor system. Calculation results are presented in Table 3. Based on Lineweaver-Burk plot and Michaelis-Menten kinetic constant calculation, constant Vmax and increased Km were obtained. Secondary metabolite produced had no effect on enzyme-substrate complex breakdown reaction into product, thus Vmax was constant. In the other hand, Km reflected affinity or binding rate of enzyme to substrate, higher Km indicated lower affinity. Analysis on Km in this study showed that the metabolite produced by L17 was competitive inhibitor. Competitive inhibitor is the the type of inhibitor which compete with substrate to bind to active site of enzyme, and unable to changed by enzyme once bound (Lehninger, 1990).

Competitive inhibition occurs due to the similar chemical structure between inhibitor and substrate, known as substrate analog (Murray et al., 2009). Characteristics of the inhibition are reversible, can be overcome by increasing substrate concentration. Inhibitor can be reversibly bound to enzyme to form enzyme-inhibitor (EI) complex instead of EnzymeSubstart (ES). However, inhibitor can not be catalyzed by enzyme into product (Lengninger, 1998). Several bacteria living and associate in host organism are able to produce various metabolites similar to the host (Tan & Zou, 2001). Ganoderma Lucidum reportedly contain high polysaccharides, with those extracted from body, spore, and mycelium of lingzhi known as Ganoderma Lucidum polysaccharides (GL-PSs) (Bao et al. 2001), triterpenoids and glycoproteins (Ma et al., 2015) whose among bioactivity are anti-inflammation, antihyperglycemia, anti-tumor and immunemodulator.

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Figure 1. Lineweaver-Burk Plot of L17 α-glucosidase inhibition kinetic measurement Table 3. Parameters Michaelis-Menten kinetics

3.4 Identification of selected α-glucosidase inhibitor-producing LAB isolate Identification was carried out using profile matching between LAB isolate L17 with several LAB references. Isolate L17 shape was identified as coccus, in pairs (diplococci), short-chained, without tetrad, able to grow at 30°C with optimum temperature at 37°C, gram-positive, catalasenegative, did not form endospore and non-motile. L17 isolate was known to be able to ferment lactose and mannitol (API 50 CHL) but unable to do so toward raffinose. Further examination was conducted through literature search using Bergey‟s Manual Determinative of Bacteriology and genus Lactococcus as reference. Characters of Lactococcus used for profile matching were: facultative anaerobic, circular in-pairs cellular shape (diplococci), short-chained, non endosporeforming, gram-positive, catalase-negative and able to grow at 30°C. The result confirmed description from Bergey‟s Manual Determinative of Bacteriology profile matching that L17 isolate was estimated to be Lactococcus. 4. Conclusion According to the results of present study, there were 14 lactic acid bacteria isolates isolated from Ganoderma lucidum, all of which able to inhibit αglucosidase activity. LAB isolate L17 was able to

produce compounds with highest inhibition activity, classified as competitive inhibition. The compound had potential to be developed into alternative antidiabetic treatment agent. Cellular identification by profile matching indicated that L17 isolate was Lactococcus. References [1] Adebayo, B. C. & Onilude, A. A. 2008. Screening of Lactic Acid Bacteria Strain Isolated from Some Nigerian Fermented Foods for EPS Production. World Applied Sciences Journal. Vol 4(5): 741-747. [2] Bachhawat, A., J, Mohamed, J. S. S. & Thirumurugan, K. 2011. Screening of fifteen indian ayurvedic plants for alpha-glucosidase inhibitory activity and enzyme kinetics. International Journal of Pharmacy and Pharmaceutical Sciences. Vol 3. [3] Bao, X., Fang, C. L. J. & Li, X. 2001. Structural and immunological studies of a major polysaccharides from Ganoderma lucidum (Fr.) Karst. Carbohydrats. 332:67-74. [4] Chen, H., Yan, X. Lin, W., Zheng, L. & Zhang, W. 2004. A new method for screening αglucosidase inhibitors and application to marine microorganisms. Pharmaceutical Biology. 42: 416-421.

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[5] Chen, Y. S., Yanagida, F. & Shinohara, T. 2005. Isolation and identification of lactic acid bacteria from soil using an enrichment procedure. Letter in Applied Microbiology. 40: 195200 [6] Coombs J. T. & Franco, C.M. 2003. Isolation and identification of Actinobacteria from surface– sterilized wheat roots. Applied Environmental Microbiology. 69: 5603-5608. [7] Guyton, Arthur C., and Hall, John E. 2007. Buku Ajar Fisiologi Kedokteran Edisi 11. EGC. Jakarta. [8] Lehninger. 1990. Dasar-dasar Biokimia. Jilid 1. Erlangga. Jakarta. [9] Loranza, B. 2012. Uji Penghambatan Aktivitas Enzim Alfa-Glukosidase dan Identifikasi Golongan Senyawa Kimia dari Fraksi Teraktif Daun Buni (Antidesma bunius L.). Skripsi. FMIPA UI. [10] Murray, R. K., Daryl K. G. & Victor W. R. 2009. Harper‟s Biochemistry. 27th. Penerbit Buku Kedokteran EGC. Jakarta [11] Pujiyanto, S. & Ferniah, R. S. 2010. Aktivitas Inhibitor Alpha-Glukosidase Bakteri PR-3 yang Diisolasi dari Tanaman Pare (Momordica charantina). BIOMA. 12 (01): 1-5. [12] Pujiyanto, S., Lestari, Y., Suwanto, A., Budiarti, S. & Darusman, L. 2012. Alphaglucosidase inhibition activity and characterization of endophytic actinomycetes isolated from some Indonesian diabetic medicinel plant. International Journal of Pharmaceutical sciences. Vol 4. [13] Putri, W. D. R., Haryadi, Marseno, D. W. & Cahyanto, M. N. 2012. Isolasi dan karakterisasi bakteri asam laktat amilolitik selama fermentasi growol, makanan tradisional indonesia. Jurnal Teknologi Pertanian Vol. 13 (1): 52-60. [14] Ramchandran, L, & Shah, N. P. 2008. Proteolytic profiles and angiotensin1converting enzyme and alpha-glucosidase inhibitory activities of selected lactic acid bacteria. Jurnal of Food Science 73(2): M75M81. [15] Shaheen, F., Ahmad, M. Khan, S. N., Hussain, S. S., Anjum, S., Tashkhodjaev, B., Turgunov, K., Sultankhodzhaev, M. N., Choudhary, M. I. & Rahman, A. 2006. New α-Glucosidase Inhibitors and Antibacterial Compounds from Myrtus communis L. European journal of organic chemistry. 2371–2377 [16] Shinde, J., Taldone T., Barletta M., Kunaparaju N., Bo H. & Kuamr S. 2008. Alpha glucosidase inhibitory activity of Syzygium cumini (Linn) Skeel seed kemel in vitri and in

[17]

[18]

[19]

[20]

[21]

[22]

GotoKakizaki (GK) rats. Carbohydrate Research. 343: 1278-1281. Simarmata, R., Sylvia, L. & Harmastini. 2007. Isolasi mikrobia Endofitik dari Tanaman Obat sambung Nyawa (Gynura procumbens) dan Analisis Potensinya sebagai Antimikrobia. Berkala Penelitian Hayati: 13 (85–90). Strobel G, & Daisy, B. 2003. Bioprospecting for microbial endophytes and their natural products. Microbiol and Moleculer Biology Review. 67:491-502. Strobel, G., Daisy B. & Castillo, U. 2005. The Biological Promise of Microbial Endophyte and Their Natural Product. Plant Pathology Journal 4(2): 161-176 Sudha, P., Zinjarde, S. S., Bhargava, S. Y., & Kumar, A. R. 2011. Potent a-amylase inhibitory activity of Indian Ayurvedic medicinal plant. BMC. Complementary and Alternative Medicine 11:5,2 Sugiwati, S., Setiasih, S. & Afifah, E. 2009. Antihyperglycemic activity of the mahkota dewa [Phaleria macrocarpa (scheff.) boerl.] leaf extracts as an alpha-glucosidase inhibitor. Makara Kesehatan Journal. 13 (2): 74-78 Wehmeier, U. F. & Piepersberg, W. 2004. Biotechnology and mollecular biology of the αglucosidase inhibitor acarbose. J Appl Biotechnol 63:613-625.

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Sari Tempe Formulation from Local Soybean (Glycine max) and It’s Sensoris and Nutritional Characteristics Muhamad Kurniadia*, Martina Andrianib, Mukhamad Angwara, Yuniar Khasanaha, Deviy Novitasary Sukamtab a) Technical Implementation unit for Development of Chemical Engineering Processes-LIPI Yogyakarta Desa Gading,Kec. Playen,Gunungkidul,Yogyakarta, 55861 Tel.:+62-274-392570; fax: +62-274-391168. b) Jurusan Studi Ilmu dan Teknologi Pangan Fakultas Pertanian, Universitas Sebelas Maret Surakarta, Jl.Ir.Sutami 36 A, Surakarta 57126 E-mail address: [email protected] Abstract : Tempe as the fermentation product from soybean has a high nutritional content such as protein, vitamin B and antioxidant. Potential products of tempe from local soybean (Anjasmoro variety) is sari tempe. Objective of this study was to determine the best formulation in the manufacture of sari tempe and its influence on sensory and chemical characteristics. The experimental design used for sensory testing was completely randomized design (CRD), which consists of two factors, variation of the ratio of water addition and tempe (1: 3, 1: 5 and 1: 7) and variations in concentrations of CMC (0.05%; 0.10% and 0.15%). Each treatment was carried out two times replicates sample and 3 repetitions analysis. Data were analyzed statistically by one-way ANOVA, if there is a real difference then continued with Ducan Multiple Range Test (DMRT) at α = 0.05. The results showed that sari tempe from local soybean with the ratio of water addition at 1: 3 and CMC concentration of 0,10%. Ratio of the water addition 1: 3, 1: 5 and 1: 7 affect the color and flavor, while CMC concentration at 0.05%, 0.10% and 0.15% did not affect the viscosity. The processing of sari tempe effect on the content of folate and vitamin B12. Keywords:sari tempe, CMC, sensory characteristics, Vitamin B 1. Introduction Tempe is a fermented product that has been known in Indonesia and consumed by almost all levels of society in Indonesia, especially in Java and Bali. Generally, tempe is made from soy, but also can be made from other materials such as velvet bean, Leucaena leucocephala, okara and the others beans. National Standardization Agency of Indonesia (BSN) (2009), define soybean tempe is a fermented product obtained from soya beans by using Rhizopus sp., Compact solid form, white slightly grayish and distinctive smell tempe. Fungi that play a role in the fermentation is R. oligopsorus and other fungi such as R. stolonifer and R. arrhizus (Kasmidjo, 1990). Vitamin B in Tempe, including vitamin B1 (thiamine), B2 (riboflavin), pantotent acid, folic acid, nicotinic acid (niacin), vitamin B6 (piroksidin) and B12 (Astawan, 1991). During fermentation, activity of vitamin B12 increased to 33 times, up about 8-47 times for riboflavin, 4-14 times for pyridoxine, 2-5 times for niacin, 4-5 times for folic acid, 2 times for pantothenic acid (Steinkraus, 1983). During fermentation, ribloflavin and biotin produced by Rhizopus oligosporus, while the folate biosynthesis by Rhizopus (Kasmidjo, 1990). Sari tempe is food products diversification of tempe (Astuti et al., 2006). Sari tempe is a second generation product of tempe besides flour, baby porridge, biscuits, ice cream, burgers, sausages and other products that has not a distinctive shape tempe

(Hapsari, 2013). This product has a high nutritional, so it can be developed into a functional beverage with a protein content of 2.5-3.0%. The process of making sari tempe is not difficult, so this product has the potential to be developed at the level of SME‟s. Sari tempe has an advantage that practically consume, suitable for lactose intolerance and non cholesterol. Manufacturing process of sari tempe using minimal heat so the nutrition can be maintained. However, sari tempe produced is usually easy to the instability caused by the emulsion due to reduced soybean lecithin during the process of fermentation (Susilowati and Aspiyanto, 2004), so the sari tempe-making in this study is necessary to add stabilizer in order to produce good quality. Carboxymethyl cellulose (CMC) is a stabilizer that serves to prevent deposition (Astawan, 2009). The combination of the addition of water and concentration of stabilizers will produce good sari tempe (Hapsari and Saihullah, 2013). Thus, it is necessary to do research on the formulation of making sari tempe from local soybean which is expected to create sari tempe which good quality in nutritional and sensory characteristics. The purpose of this research is to know the best formulation sari tempe and its influence on sensory and nutritional characteristics of sari tempe from local soybean.

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2. Material and Methode 2.1 Material Local soybean (Anjasmoro, var) from Wonosari, Yogyakarta, yeast "Raprima", plastic bags used in making tempe. Sugar "Gulaku", CMC "Point", and a plastic cup used in making sari tempe. Solvent used in analysis: HCl, H2SO4, NaOH, H3BO3, BSA (Bovine Serum Albumin), reagent Lowry A, reagent Lowry B, Na2CO3, sodium K-Tartat, CuSO4, petroleum ether, acetic acid, Pentane sulphonic.acid, Hexane sulphonic acid.sodium salt, methanol 25%. 2.2 Instrumentation Instrumentation used in this research are analytical balance (Ohaus), UV-Vis spectrophotometer Dynamica Holo RB-10, soxhlet, desiccator, analytical balance (Ohaus), oven (Memmert), mufle furnace, ultrasonic, HPLC with μ column Bondapa CN waters, UV detector. 2.3 Methodology This research was devided two stages, tempe making and sari tempe formulation and it‟s characterization (sensory and nutritional).

2.3.1.Tempe The basic method used to make tempe refers to Kasmidjo (1999) and Cahyadi (2006) to be modified. Soybean soaking with water (ratio 1: 3) for 24 hours, then drained. Soybean boiled for 30 minutes, stripping the skins. Soybeans that have been peeled and soaked again (soaking II) for 24 hours, washed and steamed for 60 minutes. Soybean cooled (40 °C) and inoculation using yeast “Raprima” (2 g/kg). After inoculation, soybean wrapped in plastic bags and incubated at room temperature for 36 hours. 2.3.2 . Sari Tempe Methods for Sari tempe making refers to Surya (2011) and modified. Tempe is cut to the shape of cubes ± 2 × 2 cm and carried steam for 10 minutes. Tempe extracted with water, filtrate obtained was diluted with water with a predetermined formula (Table 1). The filtrate was heated to a temperature of 90 ° C for 5 minutes and allowed to cool. When the temperature 50°C, add sugar and CMC (carboxymethylcellulose). Sari tempe is packed into a plastic cup and pasteurized at a temperature of 63oC for 30 minutes.

Table 1.Sari tempe formulation

2.3.3. Sensory evaluation Sensory evaluation using Test Scoring methods with 20 untrained panelists (Kartika, 1988). The parameters observed were colour, aroma, taste, aftertaste, consistency, and overall. The best results of sensory evaluation will be performed in nutritional evaluation.

(Sudarmadji et al., 2010), fat (Soxhlet) (Sudarmadji et al., 2010), soluble protein (Lowry Follin) (Apriyantono et al., 1989), total solids (Oven) (SNI, 1995 ), the levels of Vitamin B6 (HPLC) (Rooche, 1992), folic acid (Rooche, 1992) and vitamin B12 (Rooche, 1992). 3. Result and Discussion 3.1 Sensory characteristics

2.3.4. Nutritional evaluation Nutritional evaluation includes: water content (Thermogravimetri) (Sudarmadji et al., 2010), ash (Thermogravimetri) (Sudarmadji et al., 2010), carbohydrates (by Difference) (Sudarmadji et al., 2010), total protein (Semi- Micro-Kjeldahl)

Colour Sari tempe has bright yellow colour. The yellow color is due to the content of isoflavone. Flavon comes from flavus which means it is yellow (yellow), which is the colour of the majority of the flavonoids. Thus, the yellow color can be used as an

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early indicator of the presence of isoflavone compounds (Abdullah, 2009). Sensory test showed that preference for colour ranged from 2.85 (dislike) to 3.70 (like) (Figure 1). Variation ratio of water and CMC showed significant effect (p> 0.05). Panelist preference on colour F1 and F3 significantly different with F9 and F1 showed the highest.

Figure 1. Preference on sari tempe colour Note: 1 = Very Dislike, 2 = Dislike, 3 = Slightly Likes, 4 = Like, 5 = Very Likes Colour of sari tempe (F1) more interesting, bright yellow and not too muddy. Sari tempe with much water tend to be less preferred by the panelists. Sari tempe produced more pale yellow and cloudy, which is caused by the high amount of water and CMC were added. According Cahyawati (2011), a high water content in the product will cause the product looks bright because water has light-reflecting properties. Meanwhile, according to Hapsari and Saihullah M (2013), states that the concentration of the addition of carboxymethyl cellulose (CMC), which causes increasing turbidity. Aroma Panelist preference to aroma ranged from 2.85 (dislike) to 3.35 (slighty like) (Figure 2). Assessment A panelist on the parameters showed no significant difference (p> 0.05). This shows the variation ratio of the addition of water (1: 3, 1: 5 and 1: 7) as well as variations in the concentration of CMC did not affect acceptance of panelists for aroma parameters.

Figure 2. Preference on sari tempe aroma Note: 1 = Very Dislike, 2 = Dislike, 3 = Slightly Likes, 4 = Like, 5 = Very Likes

Sari tempe has a distinctive aroma of fresh tempe, which is like a mushroom aroma coming from the mycelium fungus aroma mingled with the scent of free amino acids and aroma caused by the decomposition of fat during fermentation (Astawan, 2004). Meanwhile, CMC concentration had no effect. This is consistent with the theory Tranggono (1990), which states that CMC has no taste, no smell and is a substance with a white or slightly yellowish color. Aroma may also be affected by soybean cultivars were used. According to Ginting et al (2009), Anjasmoro (var) have unpleasant intensity, so it is suitable for raw materials processed products based on soy (soy milk, tempe, tofu, sari tempe). Taste Panelist preference to aroma ranged from 2.75 (dislike) to 3.45 (slightly like) (Figure 3) and there is a significant difference (p> 0.05). Variation of the water addition (1: 3, 1: 5 and 1: 7) and CMC (0.05%, 0.10% and 0.15%) affect the panelists preference. Sari tempe (F9) deliver the highest value to the parameters.

Figure 3. Preference on sari tempe taste Note: 1 = Very Dislike, 2 = Dislike, 3 = Slightly Likes, 4 = Like, 5 = Very Likes Sari tempe has a unique taste, an unpleasant taste caused lipoxygenase enzyme activity. The presence of an unpleasant taste is a major factor in soy-based products are less acceptable (Kasmidjo, 1990). Sari tempe with greater addition of water (F7 dan F9) tend to be preferred by the panelists compared to lower (F1, F3, F5 and F6). Water addition can reduce the intensity of unpleasant taste. Formula F2 (ratio 1: 3) has a quite high and not significantly different with F9. This is due to there are some panelists who prefer sari tempe with stronger flavor of tempe. While the CMC does not affect the taste of sari tempe. This is in accordance with Tranggono (1990), CMC does not have a sense, is a substance with a white or slightly yellowish, odorless and smooth-shaped granules or powders that are hygroscopic. Aftertaste Panelist preference to aroma ranged from 2.8 (dislike) to 3.25 (slightly like) (Figure 4). Assessment panelists for the ninth aftertaste parameters soybean extract formulation showed no

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significant difference (p > 0.05). Variation ratio of the addition of water (1: 3, 1: 5 and 1: 7) as well as variations in the addition of CMC (0.05%, 0.10% and 0.15%) did not affect the panelists acceptance of the parameter aftertaste.

Figure 4. Preference on sari tempe aftertaste Note: 1 = Very Dislike, 2 = Dislike, 3 = Slightly Likes, 4 = Like, 5 = Very Likes Figure 4 shows that sari tempe (F7 and F9) have the highest value. According to the panelists, sari tempe produced have bitter aftertaste. Anjani (2013) states, that the bitter taste caused by a variety of soy compounds such as maltol, acids, palmitic, stearic, oleic, linoleic and linolenic. CMC does not effect of the parameters panelists aftertaste, because CMC has no sense (Tranggono, 1990). Formula Determination of Selected Local Soybean Tempe Sari Determination of the formula was selected based on sensory characteristics on each formulation. Sensory evaluation is very important, because food products with high quality can be eaten if not good or not appetizing because it is organoleptic properties (Soekarto, 1990). Based on sensory evaluation, can be concluded that the formulation F2 is the best formulations based on sensory testing. 3.2 Nutritional characteristics Results of the analysis carried out in selected sari tempe formula (F2) (ratio of water: tempe; 1: 3; CMC 0.10%) shown in Table 2 . Table 2. Nutritional characteristics of selected formula sari tempe Component Wet Basis (bb) Water content (%) 90,08 Ash (%) 0,08 Fat (%) 0,22 Protein (%) 1,41 Carbohydrate (%) 8,21 Dissolve protein (%) 2,74 Vitamin B6 (ppm) 20,66

Water content

Dry Basis (bk) 0,81 2,19 14,21 82,79 27,60 -

Water content of sari tempe (F2) of 90.08% (Table 2), not much different from Surya (2011), that sari tempe with honey (1: 8) in cans was 90.27%. Water addition to sari tempe will produce sari tempe with a higher moisture content. Differences ingredients such as sweeteners and stabilizers are added to increase the amount of solids, which can affect the amount of solids in water content. CMC has the properties can bind water, where the water molecules trapped in the gel structure formed by CMC, thereby increasing the total solids and water content down (Fardiaz, 1986). Thus, the larger the addition of CMC, the amount of water that is absorbed more and more so that the tendency of the water content in the solution is getting low. Ash Ash content of sari tempe was 0.81% (bk), is greater than the sari tempe produced by Surya (2011), 0.62% (db). Differences ash content can be influenced to the addition of water to sari tempe, the amount of additives added to the formulation. According Andarwulan et al (2011), the effect of processing on the material may affect the availability of minerals for the body. Minerals content from soybean extract can be donated by the Co in vitamin B12 derived from soybean. Winarno (2002) stated, vitamin B12 is a vitamin that is very complex molecule, which in addition to containing the elements of N also contains an atom of cobalt (Co) are bound similar to the hemoglobin bound iron or magnesium in chlorophyll. Fat Content Fat content in selected sari tempe was 2.19%, lower than tempe. The processing of soybean extract which involves heat causing damage and the amount of fat decreases. This is in accordance with the opinion of Palupi et al (2007), the rate of fat breakdown varies depending on the temperature used and the processing time. The higher the temperature used, the breakdown of fat will increase. Fat in soybean juice is considered better because the form of fatty acid that has sufficient solubility in water compared with other types of fat. Fatty acids in the soybean extract is indicated in the form of unsaturated fatty acids. Because of the small amount of fat and it is easily soluble in water then causes the appearance is not the same as soy milk and cow's milk (Abdullah, 2009). Protein and Dissolved Protein Protein content of ari tempe (F2) as 14.21% (db), whereas the soluble protein content of 27.60% (db). The addition of water extraction can reduce total protein sari tempe, but increased of soluble protein and cause the protein content in the raffinate (dregs) less. Protein molecule has several group consisting of N or O atoms are not paired. N atoms in the peptide chain so as to attract the negatively charged

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H atoms from positively charged water. The bound water molecules can bind with other water molecules, as it has an O atom with unpaired electrons. In addition, the protein content of sari tempe may be influenced by cooking. In the processing of sari tempe involves heat, such as boiling the extract, and pasteurization. These processes can reduce the protein. According to Winarno et al (2008), the treatment uses heat to food proteins can lead to damage such as occurs clotting protein. The occurrence of protein denaturation will cause reduced solubility. The inner layer of protein molecules that are hydrophobic would turn out, while the hydrophilic exterior will be folded into. As a result, the protein will clump and settle. Temperatures begin the denaturation of proteins occurs mostly around 70-75oC (Ophardt, 2003). Carbohydrate Carbohydrate content of sari tempe (F2) was 82.79% (db). The carbohydrate content is also influenced by the use of sugar as a sweetener. According to Winarno (2008), cane sugar as sweeteners is a disaccharide carbohydrate species are often used in the manufacture of food or drink. Vitamin B6 Vitamin B6 in the soybean extract amounted to 20.66 ppm. Vitamin B6 in the soybean extract can be influenced by the materials and processing. Tempe as the raw material in sari tempe making contributed most vitamin B6. This is because the levels of vitamin B6 tempe increased during fermentation, as a result of metabolism of mold, especially with isolates Rhyzopus oligosporus (Keuth and Bisping, 1993). Pyridoxine activity increased by 4-14 times in the process of fermentation (Astawan, 2004). Furthermore, the processing there is the process of adding water extraction and some processes involving heat, such as steaming, boiling and pasteurization. The combination of heating and addition of water in large quantities able to reduce the content of vitamin B6. Bognar (1993) states the total loss during cooking on a small amount of water will lower 211% of cooking with large amounts of water. Folate and Vitamin B12 Tempe contains many B vitamins, such as vitamin B6, folate and vitamin B12. Analysis of the content of vitamin B6, B12 and folate on the sari tempe using local soybean are shown in Table 3 Table 3. Vitamin B6, B12 and Folate on sari tempe (F2) from local soybean Vitamin Folat Vitamin B12

Tempe (ppm)

Sari Tempe (ppm)

5188,59

817,80

23,32

12,72

During the fermentation of soybeans into tempeh, folic acid content has increased 4-5 times (Steinkraus, 1983). While based on research results Vault (2013), the fermentation process can increase folic acid levels of 1.6 times. Based on Table 3, it can be seen that the levels of folic acid from local soybean (Anjasmoro, var) 5188.585 ppm , while the levels of folic acid selected from sari tempe (F2) 817.8 ppm. Decreased levels of folic acid from sari tempe as much as 6.3 time. Decreased levels of folic acid due to the addition of water, where water added can reduce the nutritional value contained. Reactivity and solubility of folic acid to water can cause loss of folic acid levels during the cooking process. A decrease in folic acid content can also be accelerated by exposure to oxygen and light during processing. Folic acid is very sensitive to sun light, storage at room temperature and normal cooking can cause a lot of folic acid is lost (Winarno, 2008). The processing of sari tempe involving a fairly high heat such as steaming tempe (for 10 minutes) and boiling the sari tempe (90 ° C for 5 min) could be expected to trigger a loss of folic acid content. In addition, folic acid found in soybean and sari tempe is a natural folic acid, which the FAO (2001) states that natural folate have a lower stability than synthetic folic acid or folic acid fortification available results, because it can retain its biological activity. Natural folate instability generated by the destruction of its biological activity when stored, processed and prepared. Vitamin B12 is highly complex, containing one atom of cobalt. Sianokobalamina (C63H88O14Pco) is the most active form of vitamin B12. Cyanocobalamin have the nature of a water-soluble, resistant to heat, inactive to light, harsh acids or lye and only a little lost by way of normal ripening (Winarno, 2008). Tempe is a potential source of vitamin B12 from food vegetable. Vitamin B12 is produced by bacterial contaminants such as Klebsiella pneumoniae (Naidoo, 1985). The fermentation process soybeans into soybean vitamin B12 can increase activity to 33 times (Steinkraus, 1983). Based on Table 3, it can be seen that the amount of vitamin B12 in tempe 23.32 ppm (bb) or 64.24 µg / g (bk). Vitamin B12 on tempe decreased after processed into sari tempe. The content of vitamin B12 in sari tempe (F2) as 12.72 ppm or 35.04 µg / g (bk), decrease 1.8 time . A decrease in vitamin B12 is relatively small, this is because vitamin B12 is only a little lost by way of normal ripening (Winarno, 2008). Besides the cooking, a decrease in vitamin B12 disebakan by the addition of water extraction, thus lowering the nutritional value contained.

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4. Conclusion Based on the research that has been done, it can be concluded as follows: 1. Sari tempe formula from local soybean (Anjasmoro, var) chosen based on the sensory characteristic, with the ratio of water : tempe (1: 3) and 0,10% CMC. 2. Ratio of water addition (1: 3, 1: 5 and 1: 7) affect the colour and taste of sari tempe, while CMC (0.05%, 0.10% and 0.15%) did not affect the viscosity. 3. Nutritional characteristics sari tempe were: water content (90.08%), ash ( 0.81%, db), protein (14.21%, db), soluble protein (27.60%, db), fat (2, 19%, db), carbohydrate (82.79%, db) and vitamin B6 (20.66 ppm).

[6] Badan Standardisasi Nasional. 2009. SNI 013922-1995 tentang Standar Mutu Tempe Kedelai. Badan Standarisasi Nasional Indonesia, Jakarta. [7] Bognar, A. 1993. Studies On The Influence Of Cooking On The Vitamin B6 Content Of Food. Journal Bioavability‟93. p. 346-351. [8] Cahyadi, W. 2006. Kedelai Khasiat dan Teknologi. Bumi Aksara. Bandung. [9] FAO/WHO. 2001. Chapter 12: Iodine. In: Human Vitamin and Mineral Requirements. Report of a joint FAO/WHO expert consultation Bangkok, Thailand, Food and Nutrition Division, FAO Rome, p. 181-194. [10] Fardiaz, S. 1986. Mikrobiologi Pangan. PT Gramedia Pustaka. Jakarta

4. The processing of sari tempe influence the nutritional characteristics of the product. Analysis of vitamin B shows, processing soybean extract lowers folate content of tempeh tempeh into juice by 6.3 time and the amount of vitamin B12 down 1.6 time.

[11] Hapsari, T. Palupi dan M. Saihullah. 2013. Pembuatan Susu Tempe Kajian Pengaruh Lama Fermentasi Tempe dan Penggunaan Carboxymethyl cellulose (CMC). Jurnal Teknologi Pangan Vol. 5, No. 1, Juni 2013 hal 2-3.

Acknowledgements

[12] Kartika, B., P. Hastuti dan W. Supartono, 1988. Pedoman Uji Inderawi Bahan Pangan. PAU Pangan dan Gizi, Universitas Gadjah Mada, Yogyakarta.

Thanks submitted to the Indonesian science agencies on research funding, and the technicians for the cooperation and assistance during the study References [1] Abdullah, Karim. 2009. Pembuatan Susu dan Tepung Tempe Sebagai Bahan Olahan Alternatif Tempe. Skripsi S1. Fakultas Matematika dan Ilmu Pengetahuan Alam. Institut Teknologi Bandung. [2] Andarwulan, N., F. Kusnandar, dan D. Herawati. 2011. Analisis Pangan. PT Dian. Rakyat. Jakarta Anjani, N. 2013. Pengaruh Bahan Penstabil (CMC, agar, pektin) terhadap Karakteristik Fisik, Kimia, dan Tingkat Penerimaan Sari Tempe. Skripsi S1 Fakultas Teknologi Pertanian. Universitas Katolik Soegijapranata. Semarang. [3] Astawan, M dan Wahyuni, A. 1991. Teknologi Pengolahan Pangan Nabati Tepat. Guna. Akademi Prassindo. Jakarta. [4] Astawan, M. 2004. Tetap Sehat dengan Produk Makanan Olahan. Tiga Serangkai. Solo. [5] Astuti, VD, Prihatini D, Rahadiyanto E, Satwiko PA. 2006. Susu Tempe Berpotensi Menjadi Pangan Fungsional. Makalah Pertemuan Ilmiah Tahunan, Perhimpunan Mikrobiologi Indonesia, 26-27 Agustus 2006, Solo.

[13] Kasmidjo, R.B., 1990. TEMPE : Mikrobiologi dan Kimia Pengolahan serta Pemanfaatannya. PAU Pangan dan Gizi UGM. Yogyakarta. [14] Keuth, S., Bisping, B. 1993. Formation of vitamins by pure cultures of tempe moulds and bacteria during the tempe solid substrate fermentation. Journal of Applied Bacteriology 75, pp. 427-434. [15] Khasanah, Yuniar. 2013. Pengaruh Asupan Tempe Terhadap Status Folat Pada Tikus (Sprague dawley). Tesis. Program Studi Ilmu dan Teknologi Pangan. UGM, Yogyakarta. [16] Nugroho, A.; Purwadi; dan L. E. Radiati. 2014. Kombinasi Carboxy Methyl Cellulose (CMC) dengan Gel Lidah Buaya (Aloe barbandensis Miller) Sebagai Thickening Agent Terhadap Kualitas Es Krim Kefir Ditinjau dari Total Padatan, pH, Total Plate Count dan Mutu Organoleptik. Artikel Ilmiah. Univeritas Brawijaya Malang. [17] Ophardt, C. E. 2003. Protein and Its Properties. Marcel Dekker Inc., New York. [18] Palupi, N.S., F.R. Zakaria dan E. Prangdimurti. 2007. Pengaruh pengolahan terhadap nilai gizi pangan. Topik 8. Modul e-learning ENBP. Departemen Ilmu dan Teknologi Pangan, Fateta – IPB. Bogor.

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[19] Rooche. 1992. Analytical Methods for Vitamin in Food/Pharma Premixes. New York: Open University Press, Inc. [20] Soekarto. 1990. Penilaian Organoleptik Untuk Industri Pangan dan Hasil Pertanian. Bhatara Aksara. Jakarta. [21] Steinkraus, K.H., 1983. Indonesian Tempeh and Related Fermentation. dalam : Handbook of Indigenous Fermented Foods, ed. K.H., Steinkraus dkk. Marcel-Dekker Inc., NY. Hal 1-94. [22] Sudarmadji. S, Bambang H dan Suhardi. 2010. Analisa Bahan Makanan dan Pertanian. Lyberty. Yogyakarta. [23] Surya, Reggie. 2011. Produksi Sari Tempe dalam Kaleng sebagai Upaya Diversifikasi Pangan berbasis Tempe. Skripsi Faklutas Teknologi Pertanian. Institut Pertanian Bogor. Bogor. [24] Susilowati, A. dan Aspiyanto. 2004. Alternatif Pati Jagung Termodifikasi Sebagai Pengental dan Penstabil serta Pengaruhnya Terhadap Kualitas Susu Tempe Secara Hidrolisis Enzimatik. Lembaga Ilmu Pengetahuan Indonesia. Jakarta. [25] Syarief, R. 1999. Wacana Tempe Indonesia. Universitas Katolik Widya Mandala Press. Surabaya. [26] Tranggono, dkk. 1990. Bahan Tambahan Makanan. PAU Pangan dan Gizi Universitas Gadjah Mada. Yogjakarta. [27] Winarno . F. G. 2008. Kimia Pangan dan Gizi Edisi Terbaru. M-Brio Press. Bogor.

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Pineapple Peel as a Potential Source of Dietary Fiber Rima Kumalasari, Ainia Herminiati, R. Cecep Erwan Andriansyah a)

Centre for Appropriate Technology Development – Indonesian Institute of Sciences Jl. KS Tubun No. 5 Subang, West Java 41213, Indonesia. Tel: +62260-411239; fax: +62260-411239 E-mail address:[email protected]

Abstract: Pineapple peel is a wasteobtained from pineapple diversification product that still can be utilized into a product that has economic value. Potential of functional food products of pineapple peel, such as: concentrated fiber, functional drink of dietary fiber, and nata de pina. These products are composed by cellulose that contains a lot of fiber, has low calorie, and resistant to digestion and absorption in the small intestine by fermentation process in the large intestine. Consumption of dietary fiber of 30 gram per day has been shown to inhibit the carcinogenic compounds that cause of colon cancer, to lose 5-10% weight in obese patients, lower blood glucose in patients with type 1 diabetes mellitus, generate better impact on HbA1c in patients with type 2 diabetes mellitus, reduce systolic and diastolic blood pressure in hypertensive patients significantly, reduce blood cholesterol significantly in patients with lipid disorders.Dietary fiber from pineapple peel can be used as a functional food that are beneficial to the health of the human gut, because it can provide a positive effect on health when consumed regularly at the effective amount. Keywords: dietary fiber; functional drink; pineapple peel; pineapple value-added 1. Introduction Pineapple (Ananas comosusL. Merr) is a tropical fruit that can grow well in Indonesia. Pineapple fruit has a mixture of sweet, sour, and fresh depending on species or varieties, with 53% edible part of the fruit. According to the results of the National Strategic Excellence Research (2002), the acidity of the pineapple came from vitamin C and citric acid with a high pH value of3. Susanto andBudi (1998) stated that pineapple has a high and complete nutritional contents, such as: protein (0.40 g); calories (52.00 cal); fat (0.20 g); carbohydrates (16.00 g); phosphorus (11.00 mg); iron (0.30 mg); vitamin A (130 SI); vitamin B1 (0.08 mg); vitamin C (24.00 mg); and water (85.30 g). Huang et al. (2014) suggested that 70% of the pineapple production is freshly consumed and 30% is manufactured into processed products. Product diversification from pineapple that aims to provide value-added and increase its shelf life, such asin dodol, jam, chips, and pineapple sauce are utilizing pineapple fruit meat. In addition, pineapple syrup and juice are processed through the extraction of pineapple fruit meat. Pineapple peel is a by-product of the pineapple-based production that has not been utilized optimally. According to Nurhayati et al. (2014), 25-35% of pineapple peel is wasted, depending on pineapple fruit varieties, level of maturity and stripping techniques. Pineapple peel contains 4.481 kcal/kg ofenergy and 17.53% of carbohydrates (Wijana et al., 1991); sucrose, fructose, glucose (Krueger et al., 1992); 19.8% of cellulose and 11.7% of hemicellulose (Bardiya et al., 1996); and 20-30% of the fibers contained in

pineapple peel is considered as neutral detergent fiber (Emaga et al., 2011). The by-product of pineapple fruit processing still contains nutrients that will be degraded by microbes and susceptible to decay if not properly handled, also can cause water and air pollutions. Solid and liquid waste treatment installationsrequire considerable investment costs, thus further processing and utilization of by-products will reduce costs, provide value-added products and environmentally friendly. Pineapple peel can be processed as raw material of by-product that provides economic value-added, used as nata fermentation media (Kusumanto, 2013; Majesty et al., 2015), cider from mill juice of pineapple peel (Yulita, 1989), feed for broiler chicken (Ulya, 2014), alternative feed for ruminants (Prima, 2012), and substrate raw material of bioethanol production (Setyawati and Rahman, 2010). High fiber contentin pineapple peel (20-30%) as neutral detergent fiber (NDF), has the potential as a dietary fiber that contributes to the nutrient movement in the digestive tract and increase stool volume (Linder, 1992). According to Kusharto (2006), dietary fiber intake of 20-35 g per day can be obtained by eating fruits and vegetables (O'Shea et al., 2012). Shortage of dietary fiber can cause constipation and in the long term can lead to colon cancer (Linder, 1992). It is recognized by the public, with increased consumption of functional food and a healthy lifestyle (O'Shea et al., 2012). Based on this background, the potential of pineapple peel as a source of dietary fiber needs to

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be studied, which aims to provide an overview of product value-addedfor SMEs that produce pineapple productsand increase the public knowledge regarding to the importance of the dietary fiber consumption to support human health. 2. Methods The research steps were conducted as follows: (1) Surveying the potential of pineapple peel on pineapple fruit-based SMEs in Subang Regency, West Java Province. (2) Study literature of pineapple peel potential as a source of dietary fiber

3. Results and Discussion 3.1 Chemical characteristics of pineapple peel as food The chemical composition of pineapple peel has been studied by Ban-Koffi and Han (1990); Wijana et al. (1991); Bardiya et al. (1996); and Kumalasari et al. (2010) and the results showed that pineapple peel contains insoluble dietary fiber (cellulose, hemicellulose and lignin), also contains sugars that can be used as a fermentation medium for celluloseformingbacteria to produce insoluble dietary fiber. Detailed data is presented in Table 1.

Table 1. Chemical compositions of pineapple peel Parameter Water Ash Fat Protein Carbohydrate Wet fiber pH Total solids Total N Reducing sugar Volatile compounds Cellulose Hemicellulose Lignin

Pineapple peel Pineapple peel Pineapple dry basisa) weight basisb) peelc) 92.20% 0.60% 0.48% 0.02% 4.10% 10.54% 1.66% 7.80oBrix 13.65% 89.4% 14.0% 20.2% 1.5% -

Pineapple peel pulpd) 85.28% 0.45% 0.07% 2.60% 5.36% 2.76 9.43oBrix 0.42% 3.04% -

Note : a)Ban-koffi dan Han, 1990; b)Wijana et al., 1991; c) Bardiya et al., 1996; d) Kumalasari et al., 2010.

3.2 Pineapple peel potential as a dietary fiber National pineapple production in 2007 reached 2.237.858 tons (www.bps.go.id, 2009). Furthermore, Nastiti et al. (2013) stated that the national pineapple production reached 702 tons per year. Table 2 presents the biggest 5 pineapple fruit-producing areas in Indonesia. Table 2. Provinces of pineapple fruit producer in Indonesia No

1 2 3 4 5

Province

North Sumatra South Sumatra Lampung West Java East Java

Pineapple plantation area (Ha) 340 763 484 1.767 3.013

Production of pineapple per year (ton) 32.175 72.265 45.896 167.439 285.504

Source: Nastiti et al., 2013

One of the pineapple production centers in West Java Province is Subang Regency with pineapple fruit production at 254.012 tons in 2007. Over the past 4 years (2004-2009), Center for Appropriate Technology Development, Indonesian Institute of Sciences (LIPI) has developed a pineapple-based

agro-industrial products and have been socialized in four SMEs in Subang. Solid waste such as pineapple peel is one of the problems inpineapple-processed business. Yulita (1989) had developed a cider baverage from mill juice of pineapple peel. Sriharti and Salim (2007) had utilized pineapple peel as a compost. Furthermore, Kumalasari et al.(2010) utilized pineapple peel to be processedintodietary fiber functional drink. As long as fresh pineapple fruit is used by pineapple processing industries, then there is alwaysa potential of fresh pineapple peel that can be utilized. If this fresh pineapple waste is converted into the dry ingredients with a water content of 24%, then there is a potential of 143000 tons per year of dried pineapple waste (Poerwanto, 2005). The composition of pineapple waste is 40% in the forms of peel stripping, heart or core of fruit, and crown (Buckle, 1989; Abdullah, 2007), with 5% ofpineapple peel part (Noto, 2010) According to Ayala-Zavala et al. (2010), a total of pineapple waste in pineapple fruit canning industryreached 48%, including: 9.1% of heart/core; 13.5% of peel; 14.9% of crown; and 14.5% of pulp. Furthermore, Nurhayati et al. (2014) stated that 25-

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35% of pineapple peel is produced from the pineapple fruit, but depending on the pineapple varieties, level of maturity and stripping techniques. Pineapple peel still contains nutrients and potentially useful to produce new products (Upadhyay et al., 2010). The results showed that the pineapple peel still contains carbohydrates and sugar (Wijana et al., 1991; Krueger et al., 1992) which can be utilized as a fermentation medium for the acid-and-ethanolforming bacteria (Noto, 2010; Setyawati and Rahman, 2010), cellulose-forming bacteria such asActinobacillus sp (Nastiti et al., 2013) and Acetobacter xylinum (Kumalasari et al., 2010). Pineapple peel also contains bromelain enzyme that serves as a meat tenderizer (Larrauri et al., 1997). 3.3 Pineapple products

fiber-based

functional

food

Pineapple peel can be created into functional food by utilizing the dietary fiber contained in the pineapple peel. According tothe FAO study, FOSHU, and FUFOSE, the provisions of the functional food are: (1) using materials that meet the quality standards and safety requirements as well as standards and other requirements specified; (2) has health benefit that is assessed from functional food components; (3) served and consumed as food or beverages; and (4) has the sensory characteristics such as appearance, color, texture or consistency and taste that are acceptable to consumers. Functional food that can be produced from pineapple peel are:  Concentrated fiber The making ofconcentrated fiber from the pineapple peel and pulp aims to utilize the dietary fibercontained in the pineapple peel and pulp. Concentrated fiber can be used as an enrichment material in cookies and bakery products. The process of concentrated fiber making from the pineapple peel and pulp conducted by Martinez et al. (2012) was washing the pineapple peel and pulp twice using warm water (300C)with water and materials ratio of 1:1 (v/w), drying using a tunnel dryer at 600C for 12 hours, and then pulverized into220-640 µm particle size. Study conducted by Martinez et al. (2012 found that the concentrated fiber from pineapple peel has 9.36% of moisture content; 4.17% of protein; 4.53% of ash; 1.33% of fat; and 14.58% of carbohydrate. Concentrated fiber from pineapplepeel contains 75.8% of total dietary fiber (dry weight), 75.2% of water insoluble dietary fiber/IDF, and 0.6% of soluble dietary fiber/SDF. According to ViudaMartos et al.(2012), the value of total dietary fiber in concentratedfiber from pineapple peel is higher than pomegranate peel concentrate at72.7% (dry weight), lemon peel concentrate at 70.4% (dry weight)

(Ubando-Rivera et al., 2005), mango peel concentrate at 70% and guava peel concentrate at 69% (Martinez et al., 2012), but lower than passion fruit peel concentrate (81.5%), while the IDF in the pineapple peel concentrate is still higher than the mango peel concentrate at 41.5%, passion fruit concentrate at 46.0%, and guava concentrate at 57.7% (Martinez et al., 2012).  Dietary fiber functional drink Pineapple peel can be used as fermentation media for the production of crushed celullose used as the base material of dietary fiber functional drink with the addition of 2% mung bean sprouts extract as a nitrogen source, addition of sugar up to 9°brix as a carbon source, fermentation time of 4 (four) days or 96 hours, resulting in 14.15% of product yield and 6.96% of fiber content (Kumalasari et al., 2010). The production stepsare: mill juice making from pineapple peel, mung bean sprouts extract making as a nitrogen source in the fermentation process, starter adaptation by observing the effect of urea and mung bean sprouts on the growth of Acetobacter xylinum, formulation and production of dietary fiber beverage. According to Kumalasari et al. (2010), dietary fiber functional drink can be accepted by the panelists with assessment scores of “slightly like” to “like”, shelf life of 6 (six) weeks, and meet the quality requirements of juice based on SNI 01-3719-1995.  Nata de pina Nata de pina product which is composed of cellulose that is processed from the pineapple peel extract byAcetobacter xylinum can be used as an alternative dietary fiber. In order to makeAcetobacter xylinum grows optimally, it needs sugar, nitrogen, vitamins, and minerals additions (Rosario, 1982). The incubation process is conducted by placing the fermentation pan at rest for 14 days to obtain an intactnata layers. Sugar contained in the pineapple peel can be converted into cellulose (nata) through the fermentation process. Nata is a bacterial cellulose in the solid form, transparent white in color with 95% water content, slightly chewy in texture and has a firm consistency (Suryati, 1979). Nata is produced from the fermentation process by utilizing the activity of Acetobacter xylinum sp. bacteria through the conversion of media containing simple sugars and nitrogen at the appropriate pH (Suryati, 1979). According to Thiaman and Kenneth (1975; in Kumalasari, 2002),Acetobacter xylinumcan convert 19% of sugar to cellulose when grown in medium containing glucose and medium pH around 3.5-5. Kumalasari research results (2002) innata de pina product showed a yield of 46.30%,22.80 mm thickness, fiber content of 0.92%, texture of 0.0093 mm/gr.det and organoleptic assessment such as

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creamy white color, chewy texture, and preferred by the panelists. 3.4 Dietary fiber as a health-supporting agent Increasing public awareness of food products that have health benefit results in massive production and circulation of functional food products in the market, one of them is claimed to possess high fiber content. Dietary fiber can not be digested and has no nutritional value, but it is believed to have an important function in maintaining body's health and preventing disease. According to Hardinsyah and Tambunan (2004),dietary reference intakes (DRI)of fiber for adults is 19-30 grams per day, while for children is 10-14 grams per day. Dietary fiber isnecessary for the body to facilitate the nutrient movement in the digestive tract. Shortage of dietary fiber causes constipation and in the long term can lead to colon cancer (Linder, 1992). According to Waspodo (2001),during 1996-2001 in the endoscopy room of Cipto Mangunkusumo Hospital (RSCM), there were 224 cases of colon cancer,among them 50 patients with an average age of 53.8 years. Furthermore, the RSCM data indicate that during 1998-2005, there were 216 cases of intestinal problems in Indonesia with 7.95% of developing colorectal cancer (Andra, 2007). The main influence of dietary fiber consumption occurs in the large intestine. Dietary fiber that enters the large intestine will interact with the microflora, mucosal cells, and intestinal muscles. According to Linder (1992), dietary fiber can soften the stool, thus reducing pressure on the wall of the colon and accelerate feces release. Dietary fiber such as cellulose can not be digested completely by the intestinal microflora, thus it will contribute to the feces formation by increasing the proportion of stool released and influence the feces density (Gallaher, 2000). High consumption of fiber will increase feces density and shorten the transit time (Tensiska, 2008). According to Buddington (2000), dietary fiber that can be fermented has the benefit to increase the density of lactic acid bacteria and suppress the growth of Enterobactericeae such as Salmonella in the gut, while the dietary fiber that is difficult to be fermented by intestinal microflora has a good laxative effect. Study conducted by Kumalasari et al. (2010) suggested that dietary fiber functional drink has proven its effectiveness to increase the number of goblet cells in intestinal mucosal villi of constipated rat, improve the necrosis condition of constipated rat colon, and suppress mice weight gain. Figure 1 shows the mechanism of pineapple peel dietary fiber to prevent colon cancer.

According to Salmeron et al. (1997), complex carbohydrates and dietary fiber contents contribute to the low-glycemic index values that are beneficial for people with diabetes and reduce the risk of diabetes. Similar to the the study results, Chandalia et al. (2000) suggested that a high intake of soluble dietary fiber, can improve glycemic control, decrease hyperinsulinemia, and lower plasma lipid concentrations in patients with type 2 diabetes mellitus.In addition, Ryttig (2005) mentioned that the high consumption of fiber has been shown to lose 5-10% weight in obese patients, lower blood glucose in patients with type 1 diabetes mellitus, generate better impact on HbA1c in patients with type 2 diabetes mellitus, reduce systolic and diastolic blood pressure in hypertensive patients significantly, reduce blood cholesterol significantly in patients with lipid disorders. Hernawati (2012) mentioned that the dietary fiber has a role in reducing blood lipid levels to prevent hypercholesterolemia. Furthermore, Muchtadi (2013) suggested that dietary fiber may affect the digestion and absorption of lipids in the intestine, also affect the metabolism of bile acids. This suggests that dietary fiber is able to bind or entrap excess fat and release itfrom the body through feces. Soluble dietary fiber can lower cholesterol by binding to bile acids in the small intestine that causes increased fecal bile acid excretion and synthesis of primary bile acids, and increased bile acid pool (Wolever et al., 1997). Anderson et al. (1999) stated thatsoluble dietary fiber affecting bile acid metabolism through its ability to bind bile acids in the gastrointestinal tract, thus the formation of micelles becomes distracted and bile acids reabsosption is decreased. This causes the excretion of dietary fiber complex and bile acids in stool increases. Furthermore, Bouhnik et al. (1999) explained that high consumption of soluble dietary fiber will produce short chain fatty acids (SCFA), H2, CO2, and bacterial biomass in the feces increased. Study by Huang et al. (2014) showed that dietary fiber intake derived from pineapple peel of 2.5% of the daily diet of animal models can improve feces ecosystem function and reduce toxic compounds excreted by the intestinal microflora. This suggests that dietary fiber from pineapple peel can be used as a functional food that are beneficial to the health of the human gut, similar to the suggestion of the American Dietetic Association (ADA) to consume natural food that will give a positive effect on health when consumed regularly at the effective amount.

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Dietary fiber of pineapple peel

Insoluble dietary fiber

Soluble dietary fiber

Contains :  Cellulose  Hemicellulose  Lignin

Examples of product :  Concentrated fiber  Dietary fiber functional drink



Nata de pina

Mechanism ofdietary fiberin inhibiting carcinogens

DF can increase the water content in the colon, then lowering the concentration of carcinogenic compounds and finally they becoming ineffective

DFaffecting the intestinal microflora, thus carcinogenic compounds are no longer produced

DFincreases the rate of materials that pass through the large intestine, thus carcinogenic compounds will not be in contact with intestinal mucosa cells in a sufficient period of time to cause carcinogenic

Figure 1 Functional food mechanism of pineapple speel dietary fiber to prevent colon cancer 4. Conclusion Pineapple peel can be used as an alternative source of dietary fiber that supports human health. Product diversification from pineapple peel can increase the value-added and economic value. Dietary fiber from pineapple peel can be used as a functional food that are beneficial to the health of human gut. To comply with the provisions of health benefits, it is necessary to test the effectiveness of pineapple fiber-based functional food products using experimental animals. Therefore, it can be claimed that the product can contribute to improve health status as disease prevention product.

[3]

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Natural Antioxidant Activities of “Tanduk Rusa” Fern (Paltycerium coronarium) Ade Chandra Iwansyah1, Dewi Desnilasari1, Ismi S Hanifah2 1)

Indonesian Institute of Sciences, Center for Appropriate Technology Development, Subang, West Java, Indonesia 2) Politeknik Negeri Bandung, Department of Chemical Engineering, Bandung, West Java, Indonesia E-mail: [email protected] or [email protected]

Abstract:The aim of the study was to determine the potential antioxidant activity in “tanduk rusa” fern (Platycerium coronarium) extract. Evaluation of antioxidant activity of the extract of Platycerium coronarium were studied using Folin-Ciocalteu assay and DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging assay respectively. The results of total phenolics showed that extract of Platycerium coronarium with decoction for 12 hour (PCD) contained the highest of total phenolics. DPPH assay showed that IC50 of PCD (507 µg/mL) was lowest. Based on these values, PCD is found to have the highest antioxidant activity. This study provides preliminary information about antioxidant activities of Platycerium coronarium. Keywords: Antioxidant, Platycerium coronarium, “tanduk rusa” fern 1. Introduction Natural products have been considered as a source of potential medicines from the beginning of 21st century. The extracting process of the phytochemical components of the plants grows rapidly in this decade (Mandal et al. 2007). Currently, there is great interest in finding antioxidants from natural sources to minimize oxidative damage to cells. Oxidative stress has been linked to major chronic health problems like cancer, heart diseases, food deterioration, cancer, aging, atherosclerosis, ischemic injury, inflammation and neurodegenerative health conditions such as Parkinson‟s and Alzheimer‟s diseases. Oxidative damage is caused by free radicals and reactive oxygen species, mostly generated endogenously. Free radicals are atoms or groups of atoms that have at least one unpaired electron, which make them highly unstable and reactive. Living organisms accumulate free radicals through both normal metabolic processes and exogenous sources. Although free radicals have beneficial effect during energy production and as antibacterial, excessively high levels of free radicals cause damage to cellular proteins, membrane lipids and nucleic acids, and eventually cell death (Agbo et al. 2014). A study explained about the specific categorizations of the plants as the medical plant has many kind of antioxidants compounds, such as phenolics and flavonoids. Phenolic compounds are a class of low molecular weight secondary plant metabolites. Most of these compounds are able to scavenge free radicals such as those produced during cell metabolism (reactive oxygen species (ROS) or free radicals such as hydrogen peroxide, hydroxyl radical and singlet oxygen) that can lead to oxidative stress. Special attention has been paid to plants because they are very rich sources of phenolic compounds (Gouveia et al. 2012). Some types of plants have

been used by hereditary for variety uses as traditional medicines. One of them is Platycerium coronarium (family Polypodiaceae), the local name of “tanduk rusa”. This fern not only has been used by generations of Indonesia people as decorative plants, but also for healing fever, inflammation of the outside uterus, irregular menstruation, ulcer, and abscesses (Tim CoData Indonesia 2000). Mostly Indonesian people consume these plants by brewing them using hot water or boiling (decoction). Information about the potential of antioxidant and bioactive compounds of Platycerium coronarium is still limited and scarce. Related to antioxidant that derived from plants for medicinal purposes, the extraction process is the basic stages in acquiring these compounds and an important stage in the separation and evaluating the bioactive components of variety plants. Generally, the extraction procedure used aprotic solvents (polar, dipolar) and nonapriotic (non-polar) in extracting the components of polyphenols of plants (HurtadoFernandez et al. 2010).Other fern in the same family in Polypodiaceae, Pityrogramma calomelanos have been reported contains secondary metabolite, i.e, 2',6'-dihydroxy-4'-methoxy-dihidrokalkon, kaempferol and quercetin (Suyatno 2010). Therefore, this study aims to determine the potential antioxidant activity of “tanduk rusa” fern (Platycerium coronarium) using a variety of extraction methods and solvents. 2. Materials and Methods 2.1 Materials The “Tanduk rusa” fern (Platycerium coronarium), distilled water, gallic acid, ascorbic acid, DPPH (1,1-diphenyl-2-picrylhydrazyl) was purchased from Sigma-Aldrich, sodium carbonate (Na2CO3), FolinCiocalteu, methanol and ethanol obtained from BrataChemBandung, boric acid (H3BO3), H2SO4 and NaOH.

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2.2 Methods 2.2.1 Preparation of samples The “Tanduk rusa” fern was rinsed to remove debris, dried and cut into small pieces. Macerate 100-200 grams of “Tanduk rusa” fern in distilled water or ethanol or other variable about 1 L for 24 hours (PC). Then, it got further extraction using a variety of solvents and methods as follows: water solvent (reflux for 6 hours (PCR), decoction (PCD), maceration for 48 hours (PCM1), and maceration for 24 hours (PCM2)); solvent variable (PCMet); ethanol 95% (PCEt1) and ethanol 80% (PCEt2). The extract was filtered using a 0.45 µm filter paper and ready for analysis. 2.2.2 Analysis Nutritional composition Nutritional composition was determined by AOAC (1990) viz., moisture content using direct heating method (oven); ash content using furnace method; protein content (nitrogen) semi micro-Kjedhal; fat content using soxhlet method; crude fiber and carbohydrates using LuffSchrool Method. The energy was calculated using Attwater factor (Where: 1g of carbohydrate = 4 kcal; 1g of fat = 9 kcal and 1 g of protein =4 kcal).

using equation (2) and reported as concentration of antioxidant required for 50% scavenging DPPH radicals in a specific period of time (IC50). Samples were analyzed in triplicates.  (Ac - As) %inhibitio n =  x100 ……………….. (2)  Ac  Where:

Ac = Absorbance control or blank As = Absorbance the sample

Data and Statistical Analysis Data were first tested for normality, and then subjected to analysis of variance (ANOVA). Significant differences between mean values were determined using Duncan‟s Multiple Range test (P=0.05) following one-way ANOVA. Statistical analyses were carried out using the correlation and regression program in Microsoft Excel 2007. 3. Result and Discussion 3.1 Nutritional composition The values of nutritional composition of 100 g “tanduk rusa” fern can be seen in the Table 1. Table 1. Nutritional composition of Platycerium coronarium per 100 g

Total phenolics content Total phenolics content of “Tanduk rusa” fern was determined by the modified of FolinCiocalteu assay (Watermann & Mole 1994). To 0.1 mL PC extract or distilled water or the standard solution of Gallic acid (0-900 µg/mL) is added to 2.8 mL distilled water, 2 mL of 2% sodium carbonate and left standing it for 4 minutes. Then, 100 µL Folin-Ciocalteu was added the solutions and left standing for 30 minutes. Measurement was conducted on a Spectrophotometer Uv-Vis GENESYS 10uV at wavelength (λ) = 760 nm. Total phenolics content was calculated from a standard curve of gallic acid with equations (1) and expressed as mg gallic acid equivalent (GAE) per 100 gram fresh plant (R2 = 0.994). Samples were analyzed in triplicates. Absorbance = 0.0008 Gallic acid (µg) + 0.0265 ………(1)

Scavenging Activity (DPPH) Assay The antioxidant activity of the extracts is based on activity of the stable DPPH free radical using a method described by Kumaran and Kuranakaran (2006). To the variety concentrations of PC extract solution or blank or standard solution of gallic acid or ascorbic acid (1 mL) was added DPPH in methanol solution (3 mL, 0.004%) then left standing in the dark for 30 minutes. Measurement was conducted by using a Spectrophotometer with a wavelength λ= 517 nm. The data was calculated

* Values are mean ± SD. of triplicate

Table 1 showed that “tanduk rusa” fern has moisture content (86.1%), ash (0.9%), protein (3.3%), fat (0.4%), carbohydrate (2.6%), crude fiber (27.4%), and energy (31.4 Cal). These results indicated that the composition of “tanduk rusa” fern has most water and crude fiber. Maisuthisakul et al. (2008) reported that there‟s a positive correlation between antioxidant activity, total phenolics and ash content in some plants that‟s origin from Thailand. 3.2 Total phenolics content and antioxidant activity Phenolic is an important component in plants. Phenolics compounds have antioxidant activity due to its capability to donate a hydrogen atom from its phenolic group to free radicals compounds (Tursiman et al. 2012). Total phenolics content of “Tanduk rusa” fern was tested by modified method of Watermann and Mole (1994) using FolinCiocalteu assay. The principle of this method is oxidation and reduction reactions. Phenolics compound of the extracts will reduce FolinCiocalteu to form molybdenum blue. The forming of

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the blue colour of the molybdenum is equivalent to the ion phenoxide concentration that forms. The “Tanduk rusa” fern extract showed a concentrationability scavenging the free radical activity by scavenging DPPH and the value was expressed as the 50% effective concentration (IC50). DPPH method is based on the reduction of DPPH solution in methanol by donating the antioxidant of hydrogen atom to form a non-radical compound (DPPH-H). The effect of enhancement antioxidant capacity is followed by increasing the concentration of the plant extracts. The plant extracts reduce the stable DPPH and change the purple colour of DPPH into yellow ones. The results of total phenolics content can be seen in the Figure 1.

Figure 1. The total phenolics content of Platycerium coronarium extracts. *) Values are mean ± SD. of triplicates; GAE: Gallic acid equivalent; a>b>c>d>e, same alphabetic = no difference

Figure 1 showed that “tanduk rusa” fern extracts had total phenolics contents about 26.3 – 270.6 mg GAE/100 g fresh plant followed by a significant difference (p=0.000 <α). The Duncan test showed that aqueous extract of “tanduk rusa” fern with decoction method (PCD) (270.6 mg GAE/100 g) had the highest values of total phenolics content as followed: PCEt1, PCR, PCMet, PCEt2, PCM1, and PCM2. The capability of PCM2 extracts in scavenging free radicals (IC50) was about 20073.6 µg/mL then followed by PCM1 (6253.9 µg/mL), PCR (1252.7 µg/mL), and PCD (507 µg/mL) (see Table 2). The highest antioxidant activity of DPPH was showed by aqueous extract of “tanduk rusa” fern with decoction for 12 hour (PCD) then followed by PCR, PCM1 and PCM2. According to Moure et al (2001), phenolics components and antioxidant activity of the plants extracts is affected by the type of solvent (polar and nonpolar) and the temperature of extraction. The total phenolics content correlated with antioxidant activity in plants (Maisuthisakulet al. 2008). Extract of “tanduk rusa” fern showed the correlation between its total phenolics content and antioxidant activity (Figure 2). The relationship of

total phenolics versus antioxidant activity (1/IC50) had relation coefficient R2=0.995 (Y=8E-06 – 0.00). Table 2. Antioxidant activity of Platycerium coronarium extracts

*) Values are mean ± SD. of triplicates; AOA: antioxidant activity;GAE: Gallic acid equivalent; a>b>c>d>e, same alphabetic = no difference

Figure 2. Relationship of antioxidant DPPH activity (1/IC50) versus total phenolics content in Paltycerium coronarium extracts These results showed that 99% capability of antioxidant activity in “tanduk rusa” fern extracts was derived from phenolics component. its can be explained by an increasing of the ability of antioxidant activity (1/IC50) against total phenolics content was proportional and showed positive linear correlation between them. The results also be in accordance with studies reported that Cosmos caudatus H.B.K., Polyscia spinnata, Pluchea indica Less., Nothopanaxs cutellarius (Burm.f.) Merr, Talinum triangulare (Jacq.) Willd., Pilea melastomoides (Poir.) Bl., and Etlinger aelatior(Jack) R.M.Sm (Andarwulan et al. 2010) extracts, as the same as Labisia pumila var.alata (Yusoff & Iwansyah 2011) has a high antioxidant activity also followed by a high concentration of total phenolics content. 4. Conclusion Based on the result of this study, “tanduk rusa” fern (Platycerium coronarium) had most composition of water and crude fiber. Aqueous extract of “tanduk rusa” fern with decoction for 12 hours had total phenolic content about 270.6 mg GAE/ g fresh plant and the highest antioxidant activity (IC50) (507 µg/mL). This study provides preliminary

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information about the potential of “tanduk rusa” fern as antioxidant. So, it needs to be explored and researched relating to other potential such as, antitumor, anticancer, anti-inflammatory and others biological activities.

[10]

Acknowledgement Authors are grateful to Indonesian Institute of Science (LIPI), Center for Appropriate Technology Development for supporting the material and technical. References [1] Agbo MO, Nnadi CO, Ukwueze NN, Okoye FBC. 2014. Phenolic constituents from Platycerium bifurcatum and their antioxidant properties. Journal of Natural Products.Vol: 7. pp: 48-57. [2] Alothman M, Rajeev B, Karim, AA. 2009. Antioxidant capacity and phenolic content of selected tropical fruits from Malaysia, extracted with different solvents. Food Chemistry 3: 785–788. [3] Andarwulan N, Batari R, Sandrasari DA, Bolling B, Wijaya H. 2013. Flavonoid content and antioxidant activity of vegetables from Indonesia. Food Chemistry 121: 1231–1235. [4] Association of Official Analytical Chemists. 1990. Official Methods of Analysis. 15th Ed. Association of Official Analytical Chemists. Washington, DC, USA. 12/1/2009. [5] Chan EW, Ng VP, Tan VV, Low YY. 2011. Antioxidant and antibacterial properties of Alpiniagalanga, Curcuma longa, and Etlinger aelatior (Zingiberaceae). Phcog J. 3:54–61. [6] Gouveia G, Goncalves J, Castilho PC. 2013. Characterization of phenolic compounds and antioxidant activity of ethanolicextracts from flowers of Andryalaglandulosa ssp. varia (Lowe ex DC.)R.Fern.,anendemic species of Macaronesia region. Journal Industrial Crops and Products. Vol: 42. Pp: 573–582. [7] Hurtado-Fernandez E, Gomez-Romero M, Carrasco-Pancorbo A, Fernandez-Gutierrez A. 2010. Application and potential of capillary electroseparation methods to determine antioxidant phenolic compounds from plant food material. Journal of Pharmaceutical and Biomedical Analysis 53:1130-1160. [8] Kumaran A., Kuranakaran J. 2006. Antioxidant and free radical scavenging activity of an aqueous extract of Coleus aromaticus. Journal Food Chemistry, 97:109-114. [9] Maisuthisakul P, Pasuk S, Ritthiruangdej P. 2008. Relationship between antioxidant properties and chemical composition of some

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Thai plants. Journal of Food Composition and Analysis, 21:229-240. Mandal V, Mohan Y, Hemalatha S. 2007. Microwave Assisted Extraction - An Innovative and Promising Extraction Tool for Medicinal Plant Research. Pharmacognosy Review 1: 718. Moure A, Cruz JM, Franco D, Dominguez JM, Sineiro J, Dominguez H, Nunez MJ, Parajo JC. 2001. Review: Natural antioxidant from residual sources. Food Chemistry 71:145171. Suyatno. 2010. Suatu Senyawa Antikanker dari Tumbuhan Paku Perak (Pityrogrammacalo melanos). Prosiding Seminar Pendidikan Sains. Program Pascasarjana Universitas Negeri Surabaya. Tim Co Data Indonesia. 2000. Tanaman Obat Indonesia : Tanduk Rusa (Paltycerium coronarium). Dikutip dari http://www.iptek.net.id/ind/pd_tanobat/view.ph p?id=148 , diakses pada 29 April 2013. Tursiman, Ardiningsih P, Nofiani R. 2012. Total Fenol Fraksi Etil Asetat dari Buah Asam Kandis (Garciniadioica blume). JKK 1(1):4548. Waterman FG, Mole S. 1994. Analysis of Phenolic Plant Metabolite:83 – 88. Yusoff MM, Iwansyah AC. 2011. Comparative Evaluation of Total Phenolics and Free Radical Scavenging Activity of Aqueous Extracts of Labisiapumilavar. alatafrom Malaysia and Indonesia.Proceeding of 2nd International Conference on Biotechnology and Food Science (ICBFS 2011):4-8.

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Potential of Suweg Starch HMT Modification as a Source of Resistant Starch Type III Raden Cecep Erwan Andriansyah Indonesian Institute of Sciences, Center for Appropriate Technology Development, Subang, West Java, Indonesia Email: [email protected] Abstract: Suweg starch (Ammorphophallus campanulatus var. Hortensis) can be modified by a hydrothermal process. Modifications of hydrothermal process has been done is a modification of Heat Moisture Treatment (HMT). HMT starch modification method has the potential to produce a modified starch containing resistant starch type III (Resistant Starch Type III / RS3). HMT modifications carried out at a temperature of 110 and 120 ° C with a moisture content of 20% for 16, 24 and 32 hours. Results modifications HMT obtained content resistant starch modified HMT is a 28.70% increase to 31.55%, 32.29% and 39.92% with a temperature of HMT 110 oC and 43.46%, 46.64% and 53.30% at a temperature of HMT 120 oC during HMT consecutive 16, 24 and 32 hours. Keywords: Suweg, Ammorphophallus campanulatus, modified starch, hydrothermal process 1. Introduction

2. Methods

Functional foods are foods that benefit one or more target functions in the body as well as nutrients that can strengthen the body's defense mechanisms and reduce the risk of a disease (Robertfroid 2007). According to BPOM (2005), functional food is a food that is natural or artificial has undergone the process of becoming a product or processed products, containing one or more functional components are based on scientific studies have certain physiological functions, proved to be harmful and beneficial to the current food healthy.This product has been developed in the form of food products containing probiotics and prebiotics or a combination of both in a product known as food sinbiotic (Widaningrum, 2012).

Suweg starch modification process refers to the procedure Collado et al. 2001 and Purwani et al. 2006. Process of modifications are made temperature and time of heating at a constant moisture content.

Probiotics are live microorganisms which, when it had consumed in sufficient quantities would be beneficial to its host (FAO 2007). Prebiotics are food ingridien that selectively stimulate the growth of lactic acid bacteria (probiotics) in the colon healthy human gastrointestinal (FAO / WHO 2001). RS3 potential as a source of prebiotic because it can not be digested by the digestive enzymes in the small intestine and when it reaches the large intestine can be utilized by probiotic bacteria. Another benefit of the RS3 is able to reduce the fecal fluid loss and shortens time of diarrhea in patients with cholera (Ramakhrisna et al 2000) as well as the potential to improve insulin sensitivity (Robertson et al 2005). RS3 is classed as a source of fiber (British Nutrition Foundation 2005) and is known to reduce cholesterol and the glycemic index (Lehmann et al 2002), as well as prevent colon cancer because the microflora capable of changing RS3 into fatty acid short-chain (butyric acid), to reduce stone formation bile, and helps the absorption of minerals (Sajilata et al 2006).

3. Results and Discussion Resiten manufacture of starch type III. The process of making resistant starch type III was referring to a modified method of Collado et al. 2001 and Purwani et al. 2006. The manufacturing process is 180 g suweg starch spread evenly over the baking sheet aluminum 20x8.5x2 cm3 with a thickness of 0.5 mm and a water content of starch suweg set to be 20% by means of an amount of water added to the starch suweg known water content. Furthermore, for the moisture content uniformly and homogeneously, pan containing starch suweg sealed and stored in a refrigerator (4 ° C) for 12 hours. Then put the pan into the oven Memmert Type 100-800. Process is carried out at a temperature and time. The heating time is calculated after the temperature of the material reaches the process temperature (± 47 minutes). Once the process is complete HMT, suweg starch stored at room temperature and dried for 4 hours at a temperature of 50 oC. Suweg starch modified and then sieved with a 100 mesh sieve (Retsch GmbH, Germany). Characterization is done against suweg analyzed starch pastes profile starch (RVA), starch (Sudarmadji et al. 1997) and resistant starch (Englyst et al 1992). 3.1 Modified Starches Native starch has some limitations in its use in the food industry, among other characteristics of resistance to the process of heating (heating) and stirring (shearing). Starch modification can improve the properties of the starch properties that the physico chemical properties and other functional properties. Modification of starch can be chemically

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classified according Beynum and Roels (1985) and physically. Modification of starch is physically influenced by several factors including: temperature, pressure, cutting, and water content in starch. Modified starch granules can be partial or total. The principle of physical modifications is to heating and tend to be more secure because it does not use a variety of chemical reagents. Sajilata (2006) states physical modification treatment include: extrusion, praboiling, steam-cooking, microwave irradiation, roasting, hydrotermal treatment and autoclaving. Physical modification methods that have been mentioned may increase levels of resistant starch (Sajilata et al. 2006). Steaming-cooking method and praboiling generally applied to the rice while the extrusion method is used to modify the functional characteristics of cereal starch. The modification process using high temperature, short time, and starch gelatinization occurs at a low water content. Metode of Heat Moisture Treatment (HMT) is a physical modification of starch using the principles hydrotermal-treatment method using water and heat to modify starch. HMT is done by using the amount of water content is low (<35%) and heated at temperatures exceeding gelatinization temperature. HMT methode can changed the characteristics of improving the starch gelatinization temperature, increasing the viscosity of starch paste, and increase the tendency of the starch to undergo retrogradation (Adebowale et al. 2005). Other physical treatment method is autoclaving. According Sajilata et al. (2006) heat treatment using the method of autoclaving can increase the production of resistant starch to 9%. HMT of starch modification method has the potential to produce a modified starch containing resistant starch type III (Resistant Starch Type III / RS3). In addition to potentially can increase the content of resistant starch type III, HMT modification also has the potential to decrease the digestibility of starch. 3.2 Resistant Starch Resistant starch (Resistant starch or RS) starch fermented by microflora in the colon wall, produce short chain fatty acids (short chain fatty acids or SCFA). SCFA profile obtained from the RS contains more butyrate and acetate contains less fiber than conventional food. By its nature, RS categorized as part of the dietary fiber. Resistant starch has a physiological effect beneficial to health such as the prevention of colon cancer, have the effect of hypoglycemic (lowering blood sugar levels after eating), acts as a prebiotic, reduced risk of gallstone formation, have the effect of hypocholesterolemic, inhibit fat accumulation and increase the absorption of minerals (Sajilata et al., 2006). The first type of resistant starch (RS I) consists of starches that are physically trapped in the cells of plants and food matrix, for example, coarsely ground rice. I was

influenced by a number of hospitals and treatment processes can be reduced ataudihilangkan by milling. The second type RS (RS II) composed of starch granules 10 are naturally very resistant to digestion by the enzyme α-amylase, for example on raw bananas and high amylose corn starch. The third type RS (RS III) consists of starch retrogradation formed when food containing starch is cooked and cooled. The fourth type of RS (RS IV) consists of chemically modified starches, where such modifications affect amylolytic activity of digestive enzymes (Liu et al., 2003). RS Example IV is modified starch etherification, esterification, and cross-link (crosslinking) (Bird et al., 2000). 3.3 Characteristics of Suweg Starch as Dietary Fiber Suweg starch composition has been studied by Andriansyah (2014), which presents data on the chemical composition of suweg starch native prior to modification HMT. Data details can be found in Table 1. Tabel 1. Chemical composition of suweg starch Parameter Water Ash Protein Fat Carbohydrate (by difference) Starch Resistant Starch

Suweg starch native (%) 10.62 ± 0.02 0.24 ± 0.04 0.24 ± 0.03 0.18 ± 0.01 88.72 ± 0.04 76.61 ± 0.98 28.70 ± 0.03

Andriansyah (2014) states that the results of the content of resistant starch suweg native increased after undergoing a process of HMT is 28.70% increased to 31.55%, 32.29% and 39.92% with a temperature of HMT 110 oC and 43.46%, 46.64% and 53.30% at a temperature of HMT 120 OC during consecutive HMT 16, 24 and 32 hours. Resistant starch content which is relatively large can be used as a source of dietary fiber that are beneficial to health tubuh.HMT can increase the temperature of gelatinization (Chung, 2000), limiting the swelling of starch and increase the stability of starch paste (Hoover and Vasanthan, 1994). HMT modified starch can be applied in a variety of industrial food. Aside from being a source of fiber, HMT starch can be used for the purpose of manufacture of pasta or noodles. Purwani (2006) states that vermicelli sago starch-modified raw materials HMT has a red color intensity higher but has a lower brightness when compared with vermicelli sago starch from natural raw materials. HMT modified starch ability to improve the texture of noodles is probably related to the gel strength and viscosity of the modified starch HMT. Suweg starch setback experienced by Andriansyah (2014) has a pasting characteristics as shown in Table 2.

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Tabel 2. Data of pasting characteristic native starch Parameter Pasting Temperature Peak viscosity Hot pasting viscocity Breakdown viscosity Cold pasting viscosity Setback viscosity Changed viscosity at 50 °C

Suweg starch native 82.95±0.07 oC 3958.50±45.96 cP 2799.00±38.18 cP 1159.50±7.78 cP 4617.00±53.74 cP 1818.00±15.56 cP 220.5±37.48 cP

According Andriansyah (2014), the characteristics of a setback decreased after treatment HMT modification of all treatments compared with natural starch which is at a temperature of 110 ° C for 32 hours is 331.00 ± 5.56cP and on the treatment temperature of 120 ° C for 32 hours is 168.50 ± 0.71. Setback viscosity value of various treatments HMT tends to decrease with increasing heating time and heating temperature higher than natural suweg starch. This shows suweg HMT starch has the ability to form a gel which is lower than natural starch retrogradation so it has a high ability. The ability to form a gel which shows the high stability of starch paste to the mixing process (Hoover and Manuel, 1996). According to Yadav et al. (2013), a low viscosity values setback that can be utilized in the frozen food industry and food in cans. 3.4 The Change of Resistant Starch Content as Dietaryfiber HMT modification at a temperature of 120°C with a time of 32 hours HMT able to reduce RDS value of the suweg starch native at 7.16 %. RDS value suweg starch native was 26.42 ± 0.28% and HMT due to modifications to the heating temperature of 110 ° C for 16 and 24 hours did not significantly affect the value of RDS suweg starch. However modification heating temperature HMT at 110 ° C for 32 hours and at a heating temperature of 120 ° C for 16, 24, and 32 hours a significant effect on decreasing the value of RDS starch suweg (Figure 1a). SDS is part of the starch that is converted into glucose between 20 to 120 minutes in vitro enzyme hydrolysis and the type of dominating the cereal starch raw product. The most important health benefits of SDS is related to the stability of glucose metabolism. Appropriate statement and Robin Lehman (2007), SDS has a low glycemic index medium so that the lower glycemic load than RDS food products with a high glycemic index. Therefore, it has a positive effect in preventing diabetes and coronary heart disease due to stabilize the amount of glucose in the blood. Effect of heating temperature on the concentration of starch suweg SDS is presented in Figure 1b. SDS greatest decrease in comparison with natural suweg starch (27.87 ± 0.51%) were treatment HMT 120 ° C for 32 hours (19.68 ± 3.88%) with a decrease of 8.19%.

Based on Figure 1c shows that the resistant starch modified suweg starch at 110 ° C for 16 hours significantly different than modified at 120 ° C at the same time. The condition was followed by a long time warming has 24 and 32 hours. The rate of increase in resistant starch are HMT 120 ° C for 32 hours (53.3 ± 8.67%) compared to the value of suweg starch native for resistant starch (28.7 ± 0.30%) with a value increase of 24.6%. An increase in resistant starch in starch suweg caused by the cutting straight chain of amylopectin and amylose bond formation with amylose, amylopectin or fat to form a more compact structure (Kusnandar 2010). The bond formation cause the starch is more difficult to be broken down by digestive enzymes, causing the slow release of glucose in the blood. The results are consistent with the Li et al. (2011), which make the process of HMT with 15-35% moisture content in green pea starch at a temperature of 120 °C for 12 hours where the natural green bean starch obtained 11.2% resistant starch content, after the HMT gained resistant starch content in the range of 17.1-45.2% , Furthermore, Chung et al. (2009) report the modification of HMT is done with heated 100 °C for 24 hours with a water content of 30%, resulting in resistant starch content increased to 10.5%, higher than the resistant starch content of corn starch native is 4.6%. Other studies report that resistant starch content increases from 5-25% with increasing heating time the corn starch ethanol-acid treatment after a heating HMT. HMT treatment at a temperature of 120 °C at the time 16, 24, and 32 hours showed increased levels of resistant starch at 43.46, 46.64, and 53.30% where the treatment temperature is 120 °C at the three temperatures are not significantly different. HMT can be used to increase the resistant starch content of the starch granules without damaging the structure. Increased levels of resistant starch reported by Shin et al. (2005) wherein the resistant starch is treated at HMT with a temperature of 120 °C for corn starch, peas, and lentils are respectively 7.7, 11.2 and 10.4%.

(a)

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Treatment (HMT). [Tesis]. Sekolah Pasca Sarjana. Institute Pertanian Bogor [3]

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Collado LS, Mabesa LB, Oates CG, Corke H. 2001.Bihon-type noodles from heat moisturetreated sweet potato starch. J Food Sci 66: 604-609. doi: 10.111/j.13652621.2001.tb04608.x

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Herawati Dian. 2009. ModifikasiPatisaguDenganTeknik Heat Moisture Treatment (HMT) danAplikasinyadalamMemperbaikiKualitasBi hun. Tesis. FakultasteknologiPertanian, Insitut Pertanian Bogor, Bogor.

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Hoover R and Manuel H.1996. The Effect of Heat-Moisture Treatment on The structure and Physicochemical Properties of Normal

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(c) Figure1.Effect of heating temperatures have on the levels of RDS (a), SDS (b), and resistant starch suweg (c) 4. Conclusion Methods Heat Moisture Treatment (HMT) can be used to increase the resistant starch content of starch without damaging the structure of suweg starch granula. Ontent of resistant starch increased after undergoing a process of HMT is 28.70% increased to 31.55%, 32.29% and 39.92% with a temperature of HMT 110 oC and 43.46%, 46.64% and 53.30% at a temperature of 120 oC during the time HMT HMT respectively 16, 24 and 32 hours. Resistant starch content which is relatively large can be used as a source of dietary fiber that are benefical to the health of the body. Bibliography [1]

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Andriansyah, RCE. 2014. Karakteristik Sifat Fisikokimia dan Sifat Fungsional Pati Suweg (Ammorphophalus campanulatus var. hortensis) Dengan Metode Heat Moisture

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Maizes Waxy Maize, Duel Waxy Maize and Amylomaize V Starches. J of Cereal Sci 23: 153-162 [13]

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From Local Wisdom: Preliminary Antibacterial Activity of “Tanduk Rusa” Fern (Platycerium coronarium) Dewi Desnilasari1, Ade Chandra Iwansyah1, Ria Fauziah2 1)

Indonesian Institute of Sciences, Center for Appropriate Technology Development, Subang, West Java, Indonesia 2) Politeknik Negeri Bandung, Department of Chemical Engineering, Bandung, West Java, Indonesia E-mail: [email protected] or [email protected]

Abstract: The aim of the study was todetermine the potential anti bacterial activity in “tanduk rusa” fern (Platycerium coronarium) extract. Antibacterial activity against Staphylococcusaureus and Escherichia coli was studied using diffusion method. Antibacterial activity test showed that PCEt can inhibit Gram-positive bacteria Staphylococcus aureus, but not the Gram-negative bacterium of Escherichia coli. PCEt1 had minimum inhibitory concentration (MIC)of 1-1.25% with the ratio of equity to antibiotics (Amoxicillin) was 6:1. This study provides preliminary information about antibacterial activity of Platycerium coronarium. Keywords: antimicrobial, Platycerium coronarium, “tanduk rusa” fern 1. Introduction Numerous and diverse plantswere used in the indigenous cultures of developing countries. For many people plants still form an important economic basis and are used in medicine. Plants naturally provide people with a good alternative medicine. Indigenous knowledge about the uses of plants has often been evolving for a long time through trial and error, and still is passed from generation to generation by oral tradition. Cures that are not effective at all will be selected against. This way of gaining medical knowledge does not require much understanding of how “a cure” exactly works, but although their mode of action is often unclear to science, they serve as an excellent source for developing new medicines (De Boer et al., 2006). In the last decade, the demand for antimicrobial agents is increasing due to emergent clinical microbial strains resistant to one or several antibiotics. Plants promise a source of natural antimicrobial agents. It has been reported that the antimicrobial activity of plants is related with the defense mechanism against microorganism (Kusuma et al., 2014). Platycerium coronarium (family Polypodiaceae), the local name of “tanduk rusa” fern has been used by generations of Indonesia people not only as decorative plants, but also for healing fever, inflammation of the outside uterus, irregular menstruation, ulcer, and abscesses due to infection (Tim CoData Indonesia, 2000). Mostly Indonesian people consume these plants by brewing them using hot water or boiling (decoction).Other fern in the family Polypodiaceae, Platycerium bifurcatum, methanol extract from that plant reported has antibacterial potency and related to the phenolic compund in plant (Angbo et al., 2014).

Phenolic compounds are one of the most diverse groups of secondary metabolites foundin plants. In nature they are involved in plant growth andreproduction, provide resistance from pathogens and predators and protect crops fromdisease and preharvest seed germination. It was reported that an antimicrobial action of phenolic compounds was related toinactivation of cellular enzymes, which depended on the rate of penetration of thesubstance into the cell or caused by membrane permeability changes. Increased membrane permeability is a major factor in the mechanism ofantimicrobial action, where compounds may disrupt membranes and cause a loss ofcellular integrity and eventual cell death (Cetin-Karaka, 2011).The extraction process is the basic stages in acquiring these compounds and an important stage in the separation and evaluating the bioactive components of variety plants. Generally, the extraction procedure used aprotic solvents (polar, dipolar) and nonapriotic (non-polar) in extracting the components of polyphenols of plants (Hurtado-Fernandez et al., 2010).The information about the potential of antibacterialofPlatycerium coronarium was still limited and scarce. Therefore, this study aims to determine the potential antimicrobial activity of “tanduk rusa” fern (Platycerium coronarium) using a variety of extraction methods and solvents. 2. Materials and Methods 2.1 Materials The “Tanduk rusa” fern (Platycerium coronarium), distilled water, Gallic acid, sodium carbonate (Na2CO3), Folin-Ciocalteu, methanol and ethanol obtained from BrataChemBandung, boric acid (H3BO3), H2SO4,NaOH, Nutrient Agar (NA), Nutrient Broth (NB), Plate Count Agar (PCA), Buffered Peptone Water (BPW), Mannitol Salt

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Phenol Red Agar (MSA), Eosin Methylen Blue Agar (EMBA), Difco Lauryl Tryptose Broth, MR-VPBroth, Simmons ariabl agar, and Bactotryptone. Gram-positive bacteria (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli) were obtained from the collection of the Microbiology Laboratory, Development Center of Appropriate Technology-LIPI. All reagents are for analysis. 2.2 Methods 2.2.1 Preparation of samples The “Tanduk rusa” fern was rinsed to remove debris, dried and cut into small pieces. Macerate 100-200 grams of “Tanduk rusa” fern in distilled water or ethanol or other variable about 1 L for 24 hours. Then, it got further extraction using a variety of solvents and methods as follows; solvent variable (PCMet); ethanol 95% (PCEt1) and ethanol 80% (PCEt2). The extractwas filteredusing a0.45 µm filter paper and ready for analysis. 2.2.2 Analysis Total Phenolics Content Total Phenolics content of “Tanduk rusa” fern was determined by the modified of FolinCiocalteu assay (Watermann and Mole, 1994). To 0.1 mL PC extract or distilled water or the standard solution of Gallic acid (0-900 µg/mL) is added to 2.8 mL distilled water, 2 mL of 2% sodium carbonate and left standing it for 4 minutes. Then, 100 µL FolinCiocalteu was added the solutions again left standing for 30 minutes. Measurement was conducted on a Spectrophotometer Uv-Vis GENESYS 10uV at wavelength (λ) = 760 nm. Total phenolics were calculated from a standard curve of gallic acid and expressed as mg Gallic Acid Equivalent (GAE) per mL (R2 = 0.994). Samples were analyzed in triplicates. Antibacterial Activity Antibacterial activity of P.coronarium extracts was tested using Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli). The antibacterial activity was determined by the Agar diffusion method using paper disc (Chan et al, 2011). 1 mL inoculum was added to a sterile Petri dish 90-mm and then add sterile agar (15 mL). The extract solution (10µL) was dropped to a sterile paper disc (sized 6-mm) using micropipette and incubated at 370C for 24 hours. Amoxicillin was used as the positive control. Observation of the inhibitory activity to the bacteria was determined by measuring the diameter of the inhibition (DDH) that formed around the paper disc. Then also measure the value of the minimum inhibitory concentration (MIC) or the lowest concentration of the extract that showed the inhibition. Data and Statistical Analysis

Data were obtained and presented as the mean ± standard deviation (SD) and tested for normality. The total phenolics content is calculated using linear regression (y = ax + b). Data (average) of each variable were compared using Analysis of Variance (ANOVA). If there‟s a significantly different then do the test usingDuncan‟s Multiple Range. Statistical analyses were carried out using in Microsoft Excel 2010. The value of P was less than 0.05, it was considered significant. 3. Result and Discussion 3.1 Total Phenolics Content Phenolic is an important component in plants (Tursiman et al., 2012). Total Phenolics content of “Tandukrusa” fern was tested by modified method of Watermann and Mole (1994) using FolinCiocalteuassay. The principle of this method is oxidation and reduction reactions. Phenolics compound of the extracts will reduce FolinCiocalteu to form molybdenum blue. The forming of the blue colourof the molybdenum is equivalent to the ion phenoxide concentration that forms. The results of total phenolics content can be seen in the Table 2. Table 2. The Total phenolics content variety Platycerium coronarium extracts.

* Value is the mean ± standard deviation (SD); GAE (Gallic Acid Equivalent)

Table 2 showed that “tanduk rusa” fern extracts had total phenolics contents about 533.5 – 2134.7µg GAE/mL extracts followed by a significant difference (p=0.000 <α). Result showed that extract with etanol 95% as solvent have the highest total phenolics. According to Moure et al (2001), phenolics components of the plants extracts is affected by the type of solvent (polar and nonpolar) and the temperature of extraction. Agbo et al (2014) reported semipreparative HPLC of the EtOAc fraction of the leaves of Platycerium bifurcatum resulted in the isolation a polyphenolic glycoside and a carboxylic acid. These known compounds were identified as quercetin 3-O-β-D-glucoside and 5'-Ocaffeoyl qunic acid (chlorogenic acid) by spectroscopic. 3.2 Antibacterial Activity Phenolics compound contained in “tanduk rusa” fern has a characteristic as antibacterial. Antibacterial activity test to “tanduk rusa” fern was done by Agar diffusion method with paper disc (Chan et al., 2011). The antibacterial character of “tanduk rusa” fern using a variety of solvents such

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us methanol, ethanol 95% and ethanol 80% were tested to Grampositive bacteria (S. aureus) and Gram-negative bacteria (E.coli) (Figure 2).

Figure 2. Antibacterial activity test of Platycerium coronarium extracts (A) Staphylococcus aureus; (B) Escherichia coli 50% concentration of “tanduk rusa” fern extracts had better antibacterial activity than other concentration in inhibiting the growth of S. aureus but it‟s not for E.coli (Table 3). Table 3. Antibacterial Activity of Platycerium coronarium extracts

EC = Escherichia coli, SA = Staphylococcus aureus, na = not analyzed, (-) = clear zone * Value is the mean ± standard deviation (SD)

The results of this test showed that “tanduk rusa” fern had antibacterial activity although it‟s only for inhibiting the growth of Gram-positive bacteria (S. aureus). This was caused by the difference structure of the cell wall of both tested bacteria. Grampositive bacteria appeared more susceptible to the action of phenolic acids than Gram negative bacteria. The key basic principle on the mode of action of organic acids on bacteria is that nondissociated (nonionized) organic acids can penetrate the bacteria cell wall and disrupt the normalphysiology of certain types of bacteria that we call pH-sensitive, meaning that they cannot tolerate a wide internal and external pH gradient. Mutations of the lpxC and tolC geneson E. coli seemed to amplify the phenolic acid antimicrobial mechanisms of action against Gram-negative species. Also, the number and position of substitutions in the benzene ring of the phenolic

acids and the saturated side-chain length influenced the antimicrobial potential of the phenolicacids against the different microorganisms, but in different ways (Cetin-karaca, 2011). The value of minimum inhibitory concentration (MIC) and the equality of extract activity using amoxicillin as the antibiotic (as the comparison) were determined. MIC values were tested at the concentration 1 – 4%. The MIC value of methanol extract (PCMet) and ethanol 95% were shown by concentration extracts about 1 – 1.25%, while the value for the 80% ethanol extract (PCEt2) was about 0 – 1% due to the concentration extract PCEt2 (1%) was still shown antibacterial activity (formed an inhibition zone about diameter 6.5 mm). As the concentration of extract is low and also it will be get a low ability of the test material in inhibiting the growth of bacteria. Table 3 also showed that amoxicillin at 1:25% concentration had the capability to inhibit the growth of S. aureus as forming inhibitory zone with an average diameter about 44.5 mm. The equal value of PCEt1 extract against antibiotic is 6:1 with the equation Y=11x + 30.167. Conclusion Antibacterial activity test showed that at 95% extract ethanol (PCEt1) could inhibit Grampositive bacteria, Staphylococcus aureus, but not for Gram-negative bacteria, Escherichia coli with a minimum inhibitory concentration (MIC) at 1-1.25% and the ratio of equity for amoxicillin (the comparison) about 6:1. This study provides preliminary information about the potential of “tanduk rusa” fern as antimicrobial. So, it needs to be explored and researched relating to other potential such as, antitumor, anticancer and anti-inflammatory and others bioactive compound therein. Acknowledgements Authors are grateful to Indonesian Institute of Science (LIPI), Center for Appropriate Technology Development for supporting the material and technical. References [1] Alothman, M., Rajeev, B., Karim, A. A. (2009). Antioxidant capacity and phenolic content of selected tropical fruits from Malaysia, extracted with different solvents. Food Chemistry 3: 785–788. [2] Andarwulan N.,Batari R., Sandrasari DA., Bolling B. , Wijaya H. 2013. Flavonoid content and antioxidant activity of vegetables from Indonesia.Food Chemistry121: 1231– 1235. [3] Association of Official Analytical Chemists. 1990. Official Methods of Analysis. 15th Ed. Association of Official Analytical Chemists. Washington, DC, USA. 12/1/2009.

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[4] De Boera, H.J., Koola,A., Broberg, A., Mziray, W.R., Hedberga, I., and Levenfors, J.J. 2005.Anti-fungal and anti-bacterial activity of some herbal remedies from Tanzania.Journal of Ethnopharmacology.96 : 461–469. [5] Chan EW, Ng VP, Tan VV, Low YY. 2011. Antioxidant and antibacterial properties of Alpiniagalanga, Curcuma longa, and Etlingeraelatior (Zingiberaceae). Phcog J. 3:54–61. [6] Cetin-Karaca, H. 2011. Evaluation of Natural Antimicrobial Phenolic Compounds Against Foodborne Pathogens. University of Kentucky Master's Theses. Paper 652.http://uknowledge.uky.edu/gradschool_the ses/652. [7] Hurtado-Fernandez, E., Gomez-Romero, M., Carrasco-Pancorbo, A., Fernandez-Gutierrez, A. (2010). Application and potential of capillary electro separation methods to determine antioxidant phenolic ompounds from plant food material. Journal of Pharmaceutical and Biomedical Analysis53:1130-1160.

coronarium). Dikutipdarihttp://www.iptek.net.id/ind/pd_tano bat/view.php?id=148, diaksespada 29 April 2013. [15] Tursiman, Ardiningsih P., Nofiani R. 2012. Total Fenol Fraksi Etil Asetat dari Buah Asam Kandis (Garciniadioica Blume). JKK 1(1):4548. [16] Waterman, F.G. and Mole, S. 1994. Analysis of Phenolic Plant Metabolite: 83 – 88. [17] Yusoff MM., Iwansyah AC. 2011. Comparative Evaluation of Total Phenolics and Free Radical Scavenging Activity of Aqueous Extracts of Labisiapumilavar. alata from Malaysia and Indonesia. Proceeding of 2nd International Conference on Biotechnology and Food Science (ICBFS 2011):4-8.

[8] Kumaran A., Kuranakaran J. 2006. Antioxidant and free radical scavenging activity of an aqueous extract of Coleus aromaticus.Journal Food Chemistry, 97, pp.109-114. [9] Kusuma, I.W., Murdiyanto, Arung, E.T., Syafrizal, Kim, Y. 2014. Antimicrobial and antioxidant properties of medicinal plants used by the Bentian tribe from Indonesia.Food Science and Human Wellness, 3: 191–196. [10] Maisuthisakul, P., Pasuk, S. and Ritthiruangdej, P. 2008. Relationship between antioxidant properties and chemical composition of some Thai plants. Journal of Food Composition and Analysis, 21:229-240. [11] Mandal, V., Mohan, Y., Hemalatha, S. (2007). Microwave Assisted Extraction - An Innovative and Promising Extraction Tool for Medicinal Plant Research. Pharmacognosy Review1: 718. [12] Moure A., Cruz JM., Franco D., Dominguez JM., Sineiro J., Dominguez H., Nunez MJ., Parajo JC. 2001. Review: Natural antioxidant from residual sources. Food Chemistry71:145171. [13] Suyatno. 2010. SuatuS enyawa Anti kanker dari Tumbuhan Paku Perak (Pityrogrammacalomelanos). Prosiding Seminar Pendidikan Sains. Program Pascasarjana Universitas Negeri Surabaya. [14] Tim Co Data Indonesia. 2000. Tanaman Obat Indonesia: Tanduk Rusa (Paltycerium 1st International Conference on Appropriate Technology Development 2015, Bandung, October 5th-7th, 2015 | 108

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New Development of Phytase Enzyme through modification of Substrate and Fermentation Technology Atit Kanti and I Made Sudiana Research Center for Biology of The Indonesian Institute of Sciences, Jl. Raya Bogor Km 46, Cibinong, Bogor 16911, Indonesia E-mail : [email protected] Abstract: The use of phytase as feed supplement is getting popular. However production system of this enzyme should be optimized to reduce production cost. The objective of this study was to evaluate the ability of 10 fungi isolates i.e. Aspergillus niger (5 isolates), and Neurospora crassa InaCC F226 to produce phytase, and select best phytase producer for phytase production on coconut oil cake supplemented with rice brand in solid state fermentation. Aspergillus niger Str 3 and Neurospora sitophylla (4.6 and 3.4 unit) respectively were selected for phytase production owing to its ability to produce phytase in submerge fermentation with glucose as the main carbon sources. These potential isolates were then used for phytase production on coconut oil cake supplemented with rice brand in solid state fermentation. The effect of inoculants type, initial moisture content, and additional carbon sources were evaluated to obtain optimum condition for phytase production. Media contained coconut oil cake supplemented with rice brand at ratio of 20 to 50 % could be use for phytase production. Initial moisture content and incubation time affect phytase production. Optimum initial moisture content was about 60-70%. The relation of moisture content and phytase production was Y = -0.0574x2 + 7.4119x - 207.59 with R² = 0.7797 for A. niger and Y = -0.0574x2 + 7.4119x - 208.56 with R² = 0.7207 for N. sitophyla. Additional carbon sources especially starch at concentration of 1-2 % increased phytase production. The effect of starch on phytase production was depicted as Y= 2.4089ln(x) + 37.193 with R² = 0.9375 for A. niger Str3. and Y = 2.9987ln(x) + 35.629 with R² = 0.9218 for N. sitophylla. This work conclude A. niger and N. sithophyla were good incoculant for phytase production using formulated media contained coconut oil cake and rice brand in solid state fermentation. Keywords: Aspergillus niger Str3; coconut oil cake; Neurospora sitophylla; solid state fermentation 1. Introduction Intensive studies have been focused on how to increase nutrient absorption by monogastric animal such as poultry and swine (Selle & Ravindran 2007a). Majority of poultry feed are plant origin and contain of appreciable amount of phytate phosphorus (Vielma et al. 1998). Due to inability of monogastric animals to produce phytase then this organically bound form of phosphorus is poorly absorb in their digestive systems (Pandey et al. 2001). Phytase hydrolyze phytate phosphorus into myo-inositol and release phosphorus in the form of available phosphate to the animal (Yu et al. 2004). Phytase supplement into animal feed has double merits: increasing available phosphate, Ca, Zn and Fe for animal (Haros et al. 2005), less undigested phytate entering water body hence reducing risk for euthrophication. The use of filamentous fungi for the production of phytase through solid-state fermentation (SSF) is gaining popularity (Soccol & Vandenberghe 2003). SSF is more preferential than submerged fermentation (SmF) for industrial scale of phytase production due to less waste water generated and lower energy input, simpler fermentation media, easier to control bacterial contamination, and higher organic loading (Pandey et al. 2000). Phytase also produced by some bacteria and yeast, but the use of filamentous fungi as inoculant is more preferable in SSF systems. This

could be due to higher production yields and more acid tolerance for feed production (Graminha et al. 2008). Several SSF parameters were optimized including particle size, pH, incubation temperature, initial moisture content, aeration and inoculums size to obtain highest phytase production (Cao et al. 2007). Fungal phytase including strain selections (Mucor, Aspergillus, and Rhizopus sp.) has been intensively explored (Howson et al. 1983) however the use of agricultural waste to reduce enzyme production cost have not been intensively explored. Coconut oil cake is selected due to commonly available in traditional market and commonly rice brand were selected as media since the substrate is low cost substrate containing phytate, and has not yet fully Here we present the possibility of using mixed media contained rice brand and coconut oil cake to produce phytase. We also explored the effect of adding additional carbon sources to increase phytase production. The objective of the present study was to determine the effect of media formulation on the production of phytase by the selected phytase producing fungi as well as to optimize the various process parameters that influence the enzyme synthesis.

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2. Materials and Methods 2.1 Culture maintenance Alls strains tested were first grown in PDA medium for 120 h, at 30°C. These strains were then evaluated for their ability to produce phytase under submerge culture. 2.2 Screening for phytase production To obtain phytase producing fungi, 10 isolates representing Aspergillus niger (5 isolates), Rhizophus oligosporus (2 isolates), Mucor rouxii (1 isolates), Neurospora crassa (1 isolates), and Neurospora sitophilla (1 isolate) were grown at described in session 2.1. Enrichment culture media containing 0.5 % calcium phytate as the sole phosphorus and glucose were used for the primary screening of phytase producers. The method was based estimation of phosphate solubilization from calcium phytate in aqueous media. The strains were grown under shaking condition at 150 rpm, at 30°C for 96 hours. After 96 hours of incubation time the fungal biomassa was discarded by centrifugation at 8000 rpm for 20 minutes and the supernatant was then estimated for phytase production using method described by Idriss et al., 2002. 2.3 Inoculum preparation for SSF The culture was grown and maintained on potatodextrose- agar (PDA) slants. The slants were stored at 4°C and sub-cultured fortnightly. Five-dayold fully sporulated slant was used for inoculant preparation. For this, 10 ml sterile distilled water containing 0.1% Tween-80 was added to the slant and spores were scraped with a sterile needle. The inoculant obtained contained 4.7 x · 107 spores per ml. 2.4 Substrates preparation for SSF Rice bran (RB), coconut oil cake (COC), were used as substrates for the phytase production. RB, and COC were obtained from a local market. Ten grams of the dried substrate taken in a cotton plugged 250 ml Erlenmeyer flask were supplemented with 6.0 ml of salt solution containing (%) NH4NO3 0.5, MgSO4.7H2O 0.1 and NaCl. Media for phytase production were contained percentage a mixture of COC/RB as the following: 100/0; 90/10; 80/20; 70/30; 60/40; 50/50 and 60/40. 2.5 Moisture optimation To estimate the effect of initial moisture on phytase production, the moisture was adjusted to the required level by adding distilled water. Substrates were sterilized at 121°C and 15 psi for 15 min, cooled and inoculated with 1.0 ml spore suspension (4.8 x 107 spores per ml) of fungal strain. The flasks were incubated at 30°C for 96 h unless otherwise mentioned. All experiments were carried out in 2 replicates.

2.6 The effect of additional carbon sources Five of additional carbon sources were selected to study the effect of additional carbon on phytase production. Additional carbon sources evaluated were starch, glucose and sucrose, lactose and manitol at the concentration 0.5 %. 2.7 Effect of starch concentration on Phytase production During the experiment we found starch were effective additional carbon sources to increase phytase production. Then varying starch concentration (0.5 to 2.5 %) were augmented to the best formula (RC and COC). Phytase productions were determined after 96 h incubations. 2.8 Enzyme extraction Enzyme extraction was carried out using distilled water with 0.1% Tween-80. Known quantities of fermented substrates were mixed thoroughly with the required volume of distilled water (so that the final extraction volume was 100 ml) by keeping the flasks on a rotary shaker at 180 rpm for one hour. The suspension was centrifuged at 8000 · g for 20 min and the clear supernatant obtained was assayed for phytase activity. 2.9 Phytase assay Phytase activity was assayed by measuring the amount of inorganic phosphorus released from sodium phytate solution using the method of Harland and Harland (1980). One unit of enzyme activity was defined as the amount of phytase required to release one micromole of inorganic phosphorus per minute under the assay conditions. 2.10 Protein estimation Soluble protein content of the crude samples was determined spectrophotometrically according to the method described by Lowry et al. (1951) using bovine serum albumin as standard. 2.11 Biomass estimation Fungal biomass estimation was carried out by determining the N-acetyl glucosamine released by the acid hydrolysis of chitin present in the cell wall of the fungi (Sakurai et al., 1977). For this, 0.5 g (dry wt) of fermented matter was mixed with concentrated sulphuric acid (2 ml) and the reaction mixture was kept for 24 h at room temperature (30°C). This mixture was diluted with distilled water to make a 1 N solution, autoclaved for 1 h, neutralized with 1 N NaOH and the final volume was made up to 100 ml with distilled water. The solution (1 ml) was mixed with 1 ml acetyl acetone reagent and incubated in a boiling water bath for 20 min. After cooling, ethanol (6 ml) was added followed by the addition of 1 ml Ehrlich reagent and the resulting mixture was incubated at 65°C for 10 min. Once cooled the optical density of the reaction

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mixture was read at 530 nm against a reagent blank (Blix, 1948). Glucosamine (Sigma) was used as the standard. The results obtained are expressed as mg glucosamine per gram dry substrate (gds). 3. Result and Discussion 3.1 Production of phytase by fungi The initial screening for phytase activity revealed some differences between the strains found (Table 1). Three groups can be distinguished after 96 h incubation: no activities, moderate and extensive caPhytate degradation. Highest phytase production was observed in Aspergilus niger str3. All strains Aspergillus niger tested produced phytase. The second group with moderate activities was Neurospora crassa and Neurospora sitophyla. Third group with low phytase production was Rhizophus oligosporus and Mucor rouxii. Hason et al., 1983. Eighty-four fungi from twenty five species have been examined for the production of extracellular enzymes capable of hydrolysing phytate (3-phytase, myo-inositol hexakisphosphate 3-phosphohydrolase, EC 3.1.3.8, and 6-phytase, myo-inositol hexakisphosphate 6-phosphohydrolase, EC 3.1.3.26) when grown in: (1) rapeseed meal (RSM); (2) a semisynthetic medium containing phytate as the sole phosphorus source (PSM); (3) potato dextrose broth (PDB). Although 58 active strains showed substantial activity, results in either of the media were of no value in indicating activity in RSM. There was no relationship between the ability of a fungus to hydrolyse phytate and its taxonomic position. Aspergillus ficuum NRRL 3135 had the greatest activity in the synthetic medium, and was relatively active in RSM. The extracellular enzyme had maximum activity after 10 days growth in PSM and had a temperature optimum of 55°C. Two pH optima were noted at pH 2.0 and 5.5. Inorganic phosphate inhibited enzyme production; ammonia ions were a better nitrogen source than nitrate or urea. Aspergillus niger has been intensively studied for their ability to produce phytase, and A. ficuum was proposed by Shieh & Ware (1968) to produce phytase. We found other isolates such as Neurospora crassa and N. sitophyla also produced phytase. We observed phytase was induce Caphytate. When Ca-phytase was un unavailable, Phytase was not produced (data is not shown). This verifies that phytase is inducible enzyme (Rodríguez-Fernández et al. 2012). Neurospora crassa and N. sitophyla were selected due to these fungi commonly encountered in traditional fermented food in Indonesia. Other important isolates are Rhizophus oligosporus as inoculants for tempeh production, and Mucor racemousus known as phytase producing fungi (Roopesh et al. 2006). The best idea could be phytase production system is

utilizing generally microorganism.

well

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Table 1. Phytase activity of fungi grown on glucose with 0.5 % Ca-phytate contained (g per liter) glucose 30,0; MgS04, 0.5 g; KCl, 0.4 g; FeSO4, 0.08 g; and NaNO3 8.5 g , grown under shaking condition at 30°C No.

Species name

1 2 3 4 5 6 7 8 9 10

Aspergillus niger Str1 Aspergillus niger Str2 Aspergillus niger Str 3 Aspergillus niger Str4 Aspergillus niger Str 5 Mucor rouxii Neuropora crassa Neurosphora sitophila Rhizophus oligosporus Rhizophus oligosporus

Acid phytase production (Unit) 2.6 ± 0.1 3.2± 009 4.6± 009 1.2± 009 3.9± 009 1.9± 009 3.2± 009 3.4± 009 1.4± 009 1.2± 009

3.2 Activity of Phytase in formulated media Media formulation was chosen as strategy to reduce phytase production cost. Gradient composition of coconut oil cake and rice brand were explored to achieve cost effective phytase production. Coconut oil cake was selected due to low price of this media in traditional market, and rice brand is commonly used for phytase production (Selle & Ravindran 2007b). As shown by Table 2 media composition and inoculants type affect phytase production. Aspergillus niger (Str2 and St3), and Neurospora crassa and N. sitophylla were good inoculant, and COC: RB at ratio from 30 to 60 could be used to produce phytase. Optimation of medium component to achieve higher phytase production was introduced by (Buckle 1988). Coconut oil cake inoculated with Rhizopus oligosporus in Solid-state fermentation was conducted by Sabu et al., 2002. They observed maximal enzyme production was 14.29 U/g of dry substrate, occurred at pH 5.3, 30º C, and 54.5% moisture content after 96 h of incubation. We observed that using formulated media contained rice brand and coconut oil cake was able to increase phytase production by more than 40 % (Table 2). Not only coconut oil cake, sesame oil cake were used by Singh & Satyanarayana (2006) for phytase production using Sporotrichum thermophile in solidstate fermentation. They found Sporotrichum thermophile TLR50 increased phytase production to 180 U/g of dry moldy residue in sesame oil cake at 120 h and 45° C. The the initial substrate-tomoisture ratio of 1:2.5 and aw of 0.95. Addition of carbon and nutrient sources i.e. glucose and ammonium sulfate enhanced phytase titer (282 U/g of dry moldy residue). An overall 76% enhancement in phytase production was achieved owing to optimization. Not only phytase this fungi also excreted amylase, xylanase, and lipase. While Roopesh et al., 2006 compared the phytase

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production of wheat bran and oil cake using Mucor racemosus as inoculant. To the best of our knowledge, only our study introduced formulated media containing coconut oil cake and rice brand for phytase production. Those optimation of media and microorganism in solid state fermentation are common strategy to reduce phytase production cost. There is significant different phytase production character in solid state and sub merge fermentation. Aspergillus niger NCIM 563 produce more phytase type in solid state than under submerge fermentation (Bhavsar, et al., 2013). Various oil cakes (sesame, ground nut) and mixed substrate contained those oil cake were studied by Roopesh et al., 2005.

1994. Sesame oil cake served as the best carbon source for phytase synthesis by the fungal strain as it gave the highest enzyme titres (30.6 U/gds). Groundnut oil cake also produced a reasonably good quantity of enzyme (24.3 U/gds). They found, wheat brand which commonly known as the best media for phytase production, produce much less phytase than sesame oil cake. Increased phytase production was obtained when wheat brand was supplemented with sesame oil cake (1:1). The highest phytase production (44.5 U/gds) was attained though optimizing various process parameters such as incubation time, initial moisture content and inoculum concentration. Implying that substrate formulation and environmental manipulation are crucial for phytase production.

Comparisons were made for phytase production using wheat bran and oilcakes as substrates in solidstate fermentation by Mucor racemosus NRRL Table 2. Phytase activity of fungi in formulated media 0: 100

1 2 3 4 5

Aspergillus niger Str2 Aspergillus niger Str 3 Neuropora crassa Neurosphora sitophila Rhizophus oligosporus

31.3 ±0.4 27.3±0.6 26.3±0.3 19.6±0.5 8.1±0.5

10: 90 31.5±0.3 28.6±0.3 28.5±0.6 26.5±0.5 12.5±0.5

3.3 Biomass growth, protein production and enzyme activities Enzymes production is concomitant with biomass growth and protein synthesis (Figure 3, and Figure 4) for Aspergillus niger and N. sitophylla respectively. This might suggest biomass formation require more phosphate from their surrounding, and thus stimulating phytase production (Figure 3 and 4). Enzyme production was maximum at 96 h. This result is reaffirm the previous study of (Ramachandran et al. 2005) which noted that optimum production of phytase in oil cake based media was attained after 96 hours. Protein synthesis is slightly increase which may indicate that this fungi produced other enzymes as reported by Singh & Satyanarayana (2006) who evaluate enzyme hydrolyses by Sporotrichum thermophile. Beside phytase this thermophilic fungi also produced amylase, xylanase, and lipase when gron in sesame cake oil. Production of other hydrolytic enzymes could be benefial to increase nutrient availability when mixed with animal feed. Both Aspergillus niger St3 and N. sitophylla showed similar pattern, but phytase production N. sitophyla greatly decreased after 100 h fermentation period (Figure 4). Therefore it is important to monitored enzyme production profile to obtain most efficient phytase production system.

Media composition (COC:RB) 20:80 30: 70 40: 60 (Unit) 27.5±0.2 34.2±0.2 31.1±0.3 29.8±1.5 34.2±1.4 33.2±1.3 34.5±3.4 34.2±1.4 31.1±0.3 35.5±2.2 34.2±2.3 31.1±0.5 15.5±0.6 16.2±0.4 22.1±0.5

50:50

60: 40

27.4±0.6 34.5±0.1 26.8±0.6 25.4±0.5 27.4±0.4

27.1±0.7 33.9±0.2 27.1±0.5 30.8±0.5 27.1±0.3

100

40

90

35

80

30

70 60

25

50

20

40

Biomass Protein Phytase

30 20

15 10 5

10 0

Phytase Activity (Unit/g.ds)

Species name

Biomass (mg/g.ds) Soluble protein (mg/g.ds

No.

0 0

50 100 150 Incubation Time (h)

200

Figure 3. Phytase activity, growth and soluble protein content at different time intervals of Aspergillus niger St3 in formulated media COC:RB of 30: 70 respectively 3.4 Effect of initial moisture The profile of phytase production was affected by initial moisture content of media. Both Aspergillus niger and N. sitophylla showed similar pattern. When the initial moisture contentwas higher than 70 %, then the phytase production inhibited (Figure 5). Substrate moisture is one important parameter for solid state fermentation. We found substrate moisture also affect growth of mycelia in substrate. When initial moisture was too high (>85 %) miselia grow was very slow, and media easily contaminated by other microorganisms (bacteria). When The effect of moisture content would be thorough limiting oxygen diffusion for mycelia

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growth(Papagianni et al. 1999), and due to much water media became sticky. We try several sterilization techniques to get most suitable initial moisture content for stimulating mycelia growth and hence stimulating enzyme production. The effect of initial moisture was Effect of supplementation of additional carbon sources on phytase production.

4. Conclussion Phytase could be produced optimally through substrate formulation and optimazing factor affecting fermentation process. Coconut waste could be used as good substrate for phytase production. Acknowledgements This project is funded by The Indonesian Institute of Sciences through Commercial Product Development Program 2015. The author acknowledge Dr. Lisman Suryanegara as the program coordinator, and Ms. Senlie Oktaviana and Rizka Syahputri for laboratory work. References [1]

Bhavsar, Kavita, Priyanka Buddhiwant, Sarvesh K. Soni, Dilip Depan, Sampa Sarkar, and Jayant M. Khire. 2013. “Phytase Isozymes from Aspergillus Niger NCIM 563 under Solid State Fermentation: Biochemical Characterization and Their Correlation with Submerged Phytases.” Process Biochemistry 48 (11): 1618–1625.

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Figure 5. Effect of initial moisture on phytase activity on substrate containing COC:RB of 30: 70 respectively, inoculated with Aspergillus niger St3 and Neuorospora sitophyla

Figure 6. The effect of additional carbon sources on phytase production, fermentation period (96 h, initial humidity (60 %) with COC:RB of 30: 70 respectively inoculated with Aspergillus niger St3 and Neuorospora sitophyla

Figure 7. Effect of starch addition on phytase production, at 96 h fermentation time, 60 % initial moisture content on media composition COC:RB of 30:70 respectively, inoculated with Aspergillus niger St3 and Neuorospora sitophyla

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Silicon. Journal of Applied Physics 36, 37– 70. [8]

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Graminha, E B N, A Z L Gonc, and R D P B Pirota. 2008. “Enzyme Production by SolidState Fermentation: Application to Animal Nutrition” 144: 1–22. doi:10.1016/j.anifeedsci.2007.09.029.

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Singh, B., & Satyanarayana, T. (2006). Phytase production by thermophilic mold Sporotrichum thermophile in solid-state fermentation and its application in dephytinization of sesame oil cake. Applied Biochemistry and Biotechnology, 133(3), 239–250.

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Idriss, E. E., Makarewicz, O., Farouk, A., Rosner, K., Greiner, R., Bochow, H., Borriss, R. (2002). Extracellular phytase activity of Bacillus amyloliquefaciens FZB45 contributes to its plant-growth-promoting effect. Microbiology, 148, 2097–2109.

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Mandviwala, T. N., & Khire, J. M. (2000). Production of high activity thermostable phytase from thermotolerant Aspergillus niger in solid state fermentation. Journal of Industrial Microbiology and Biotechnology. doi:10.1038/sj.jim.2900811 Pandey, A., Soccol, C.R. & Mitchell, D., 2000. New developments in solid state fermentation: I-bioprocesses and products. Process Biochemistry, 35(10), pp.1153–1169.

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[29]

Soccol, C.R. & Vandenberghe, L.P.., 2003. Overview of applied solid-state fermentation in Brazil. Biochemical Engineering Journal, 13(2-3), pp.205–218.

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Vielma, Jouni, Santosh P. Lall, Juha Koskela, Franz Josef Schöner, and Pirjo Mattila. 1998. “Effects of Dietary Phytase and Cholecalciferol on Phosphorus Bioavailability in Rainbow Trout (Oncorhynchus Mykiss).” Aquaculture 163: 309–23.

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ECONOMY, ENERGY, ENVIRONMENT, MANAGEMENT

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Interface between Food Security, Energy Sustainability and Water Accessibility Leuserina Garniati1, Radisti Praptiwi1,3, Novieta Herdeani Sari3, Yoyon Ahmudiarto2,Jito Sugardjito3, Alan Owen1 1)

Centre for Understanding Sustainability in Practice, Robert Gordon University (RGU) Riverside East, Aberdeen AB10 7GJ, United Kingdom 2) Lembaga Ilmu Pengetahuan Indonesia (LIPI), Pusat Pengembangan Teknologi Tepat Guna Jalan K. S. Tubun No. 5, Kecamatan Subang, Indonesia 3) Centre for Sustainable Energy and Resources Management (CSERM), Universitas Nasional Jalan Sawo Manila 61, Pejaten, Jakarta Selatan, Indonesia E-mail : [email protected] Summary: Indonesia, with its vast population, faces challenges of fulfilling the high demands for food and clean water. To meet the demands, it is often unavoidable that extensive use of energy for food and water production interferes with the need to conserve environmental and natural resources. The challenge to meet the food and energy demands without jeopardising the ecosystem conservation is further made more complex by the need to provide remotely located areas with sufficient power. These problems nevertheless are observed to be able to be mitigated by thinking holistically, using models incorporating the interlinked flow of food, energy and water (FEW nexus). In addition to the nexus, the involvement of the local communities is essential to help provide knowledge and feedback throughout the whole decentralised interface of the system. The local wisdom should prevent a cyclic friction between energy generation processes and the needs of protected areas . Keywords:Appropriate technology, Sustainable energy, F-E-W nexus, local needs 1. Background Indonesia, with the fourth highest population in the world, has a strong political and economic role in South East Asia. It is an emerging nation, rich in biodiversity, with a high energy demand to support its economic activities, but also unequally distributed energy in its remote regions, which often hinders the management of natural resources and land uses (DFID 2013). Since 1990, Indonesia has lost nearly a quarter of its forest that it could lose all remaining forests by 2056 to rubber, oil palm and pulp plantations, at current rate of deforestation (Conservation International 2014). Indonesia, nonetheless, has taken steps to designate its forests as protected areas to mitigate such threat and ensure that the ecosystem services from the forests can be provided sustainably. These areas are designed to fully consider the presence of local communities residing within or in the proximity of the protected areas. These communities are the primary guardians of the ecosystems, providing the buffer zones against large scale economic activities (Eghenter 2000). However, these large scale economic pressures, especially in the form of land use changes to accommodate timber, palm oil and mineral extraction industries are considered to be a threat to the co-existence between the local people and the forests. Driven by the desire for improved economic wellbeing, healthcare, and other services, those indigenous communities are often drawn into such enterprises (Oglethorpe et al. 2007).

In the next decade the exponential growth of population will be an ever increasing threat to the global food production system, water scarcity and deforestation (KPMG International 2012). Forests products contributed $100 billion per year to the global economy from 2003 to 2007 and the value of non-wood forest products (mostly food) was estimated at US$18.5 billion in 2005 (FAO 2006). Yet, approximately 40% of the world‟s natural forests have disappeared in the last 300 years (FAO 2006). It is also predicted that the world‟s forests area will continue to decline by 13 percent from 2005 to 2030, mostly in South Asia and Africa (KPMG International 2012). Both the rate of forest degradation and the loss of other types of natural ecosystems continue unabated (Butchart et al. 2010). Despite the short term economic benefits of palm oil, rubber, and pulp plantations, they also accelerate greenhouse gas emissions, increase air pollution and harm the important forest ecosystems. Worldwide cases of forests ecosystem breakdown and stress have illustrated the dependence of human livelihood and business operations on the critical services such ecosystem provides. The decline of forest ecosystems (1) causes natural resources to become scarcer and more expensive, (2) increases the costs of water, and (3) escalates the damage caused by invasive species to the agriculture, fishing and food production sectors. Based on the Indonesian Government‟s long-term strategy, the priority sectors for development in Indonesia emphasise poverty reduction and remote region infrastructure and basic food/water/energy systems development (MP3EI 2012). Indonesia‟s

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protected areas have high value as natural ecosystems and potentially can be managed in the context of The Economics of Ecosystems and Natural ecosystem (TEEB) initiative, which strongly highlights the urgent need to incorporate ecosystem services, especially forest carbon, into national accounts. Indonesia‟s newly elected government seeks to overcome this barrier through its commitment to intensify research in pertinent areas. It has committed to increase its research budget to ensure research-based policy and to use research to address key issues such as food security, the development of Indonesia‟s remote villages and sustainable energy (Republika 2014, IRRI 2015). In summary, sustainable energy and ecosystem conservation issues have appeared to be thoroughly and competently addressed by many academics. However, decentralised sustainable energy systems and natural resources management within ecosystem conservation are less frequently researched simultaneously, and often fail to address emerging/developing nations‟ multifaceted societal constraints. There have been many attempts to provide remote communities with off-grid renewable energy solutions capable of supplying a few kilowatt-hours for lighting and water pumping, but these often prove to be inappropriate in terms of both the technology and in meeting the societal and cultural needs of the community. 2. Scope of paper This paper aims to identify the interfaces between decentralised sustainable energy and ecosystem conservation; and construct the bridges necessary to address the gaps between their discrete components. To address this, information are analysed through the following logical framework:  The current state of play (Section 3) highlights the issues surrounding decentralised sustainable energy and ecosystem conservation in Indonesiaspecific situation are put in context globally  The steps to identify the interfaces between sustainable energy and natural resources conservation are explored in detail through Section 4, and the gaps identified between existing components are highlighted  Potential actions to bridge the gaps between components which extends pilots into mainstreams are subsequently proposed through Section 5 3. Current State of Play One of the immediate, main challenges confronted by rural and/or coastal communities in Indonesia relating to food security, water availability, and direct economic productivity is energy accessibility (not only electricity) for production, processing, and storing/preserving agricultural products (e.g. drying

and cooling). Many of these communities in Indonesia are located in remote areas with limited or no access to grid electricity, where fossil fuels are the current primary energy supply and even so, their availability and accessibility are relatively low. Many of these communities are also living adjacent to protected areas. It becomes imperative that alternative means of income generation exist to prevent the contribution of indigenous people to the ecosystem degradation. This can be provided by giving access to affordable energy as the primary driver of improved livelihood in these communities. Renewable sources of energy can be used to support farming processes, increasing crop yields and improving the processing, storage, and access to market. This should provide the means of addressing food security through better agriculture/aquaculture and water management. By protecting the natural forest environment and avoiding the use of fossil fuels, it also contributes to the climate change mitigation strategy. 3.1 Sustainable energy decentralisation

system

and

A sustainable energy system requires the spread of energy services to reach disadvantaged populations, implementation of rational pricing strategies, and actions for structural reform to ensure facilitation and financing of technology transfer (Saha 2003). The social component of sustainable energy can thereby be expanded to cover community involvement, affordability, social acceptability, lifestyles, and aesthetics (Rosen 2009). A fourth dimension of sustainable energy accounted for in this research is political commitment (Kruijsen et al. 2012). The sustainable energy systems discussed in this paper therefore consist of:  Sustainable energy consumption, which includes energy conservation measures;  Sustainable energy generation, which includes renewable energy provision;  Sustainable energy distribution, which includes equal and secure access to energy resources Sustainable energy resources currently identified to be readily exploitable and available in Indonesia include biomass, biogas, mini hydro, wind and solar (PV/thermal), marine current, wave, and ocean thermal with different levels of penetration and/or potential of application. This paper interprets that sustainable energy in practice as energy (not simply electricity) from renewable sources that is not, in its life-cycle, a net contributor to climate change and does not have substantially negative environmental, social, economic, and political impacts. In itself, sustainable energy in practice needs to be seen as an integrated part of a Food-Energy-Water (FEW) resource

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management through appropriate technology inovation and diffusion. This process are dependent on the balancing act between the four “P”s of sustainable practice (i.e. Politics, People (social aspect), Planet (environmental aspect), and Profit (Economic aspect). Conventionally, sustainable solutions rely on centralised capacities for energy generation which would subsequently be distributed concentrically to the areas where the generated energies will be consumed. However, such systems could present major challenges, particularly for the delivery to remote areas, due to their costs and technical inefficiencies, safety and environmental risks, and accessibility (Practical Actions n.d., Pepermans et al. 2005, South Centre 2008, Ramchurn et al. 2012). In contrast, recent developments of sustainable energy indicate the ever lessening reliance on large grids required to deliver power in a centralised system. Many methods used for generating sustainable energy do not require extensive and intensive usage of resources, such as fossil fuels, lands and technological investments (Land Use and TV Energy 2010, Calvert and Mabee 2014, McLellan et al. 2015). This enable the small scale adoption of the technologies directly within the communities requiring the energy supply for their activities, leading to decentralised energy generation systems which would benefit in higher efficiency and reduced carbon emissions (Chicco and Mancarella 2009, Karger and Hennings 2009, McLellan et al. 2015). 3.2 Energy demand, ecosystem services, and protected areas Although Indonesia is a producer of coal and crude palm oil which have been used to fulfill energy demand, the local people in the remote areas have not yet benefitted directly. Furthermore, extractive activities to fulfill energy demand, including coal mining and crude palm oil plantation, threaten the often already endangered natural forest biodiversity. These activities often neglect the holistic consideration of ecosystem, social and cultural values inherent in the areas. For example, in the case with palm oil plantation, several studies have repeatedly shown that the economic activities of palm oil production have generated detrimental ecosystem impacts in the forms of significant decline of forest and wetland areas in Indonesia (Koh and Wilcove 2008, Koh et al. 2011, Miyake et al. 2012). The destruction of such important ecosystems contributes to the acceleration of climate change through the release of greenhouse gasses and the reduction of carbon sequestration capacity (Danielsen et al. 2009, Carlson et al. 2013), and the loss of biodiversity (Koh and Wilcove 2008, Barnes et al. 2014).

Culturally, the economic activities have also resulted in land disputes with local communities, often causing these people to be removed from their lands (Naylor et al 2007, The Jakarta Post 2011). As comparison, a specific case study in Central Africa demonstrated that extractive industries could be designed and implemented with an inclusive consideration of ecosystem conservation needs (Tolisano et al. n.d.). Such integrated approach has been observed to be more readily adaptable for the various methods associated with the generation of readily available renewable energy, especially those that does not involve radical changes of local patterns of land use (Pelc and Fujita 2002, Anderson and Fergusson 2006, Omer 2008, Groom et al. 2008, Inger et al. 2009, Tilman et al. 2009, Quanz et al 2013). 4. Analysis - Identifying the interface Decentralised sustainable energy and ecosystem conservation research streams have a very limited existing interface both academically and practically. Therefore, to identify this interface, we have developed a conceptual boundary based on the current state of play in the sectors, to scope the analytical framework (Figure 1). Identification of the interface between decentralised sustainable energy and natural ecosystem conservation means that it is necessary to evaluate the existing mutually beneficial or conflicting interests in the utilisation of resources between biodiversity conservation and community development and industrial needs and aspirations. This includes: 1. Identification of what the issues are in remote communities around energy use (societal/cultural/technical and economic), the sustainable use of natural resources and the subsequent impact on conservation issues, with particular focus on:  Food-water-energy nexus around protected areas.  Industrial processes energy and land use needs.  Effective and efficient models for storing, cooking and heating food-stuff.  Harvesting system of non timber products and replanting using fast growing fuel-wood trees. 2. Determination of the energy resources available to and the existing primary energy consumption of the community and the additional activities and processes they envisage using if greater access to energy is available:  Analyse surrounding energy resources distribution, access, usage etc and potential

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conflict with neighbouring communities and biodiversity.  Measure availability and magnitude of solar, wind, hydro and biofuels available as energy resources (the equipment listed in the budget will be utilised for this purpose).  Analyse potential impact on economic outputs and unintended consequences for conservation of changes in energy empowerment.  Assess existing conservation, industry, and community energy use and understand community aspirations. It is clear that the discussion cannot escape the issues of land use and natural resources management, with appropriate technology located at

the centre of the interface between decentralised sustainable energy and ecosystem conservation. At this interface, appropriate sustainable energy technology is crucial in connecting the FEW nexus as products of ecosystem services to the decision making related to natural resources management. Appropriate technology needs to be implementable and supported by committed policy for the long term. This means that an innovative integrated model for a local business case becomes important, especially to determine the key actors and stakeholders roles (e.g. in cultural leaders, businesses, local/national governments, households, utility companies, academia, etc.) and the costs/benefits each group will be exposed to. Such model should holistically consider the presence of interlinked FEW nexus.

Figure 1: Conceptual boundaries An integrated platform representing FEW overlaps with sustainable resources management, appropriate technology innovation and diffusion, and policy framework and business case was developed based on the following overview and is represented by Figure 2. Figure 2 indicates that the three elements of water, food and energy significantly regulate the areas of resources management, and technology innovation and diffusion. Water and energy are needed to grow food, to drive the associated processing and preservation activities, and energy is required to treat and transport water. The relationships and tradeoffs within this triangle of resources iterate that food, water, and energy are inextricably interdependent. Whilst current policies and business

models often treat water, energy and food security separately, issues in one of these sectors must be addressed with the understanding of this interdependence, and seek for holistic solutions to address the FEW nexus. Existing energy generation to support agricultural activities is primarily met by diesel; however, the fuel subsidy is being phased out, increasing the vulnerability of remote rural and/or coastal communities. Preliminary discussion and informal interviews with the senior management of the Indonesian Centre for Agricultural Socio Economic and Policy Studies, Ministry of Agriculture, have identified the needs for alternative energy as part of a closed loop mechanism for the sustainability of agricultural communities.

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Figure 2: Resources-Technology-Policy & Local Business Platform for FEW Nexus in Indonesia

Figure 3: Formulation of business model and teaching/training material Therefore, as represented by Figure 3 it is imperative to formulate a local business model based on evidence based conceptual frameworks of provision and distribution of appropriate sustainable energy (e.g. micro hydro, biomass, palm oil waste, solar, marine current, wave, and ocean thermal etc.) through amongst others: 1.

Determination by how much industrial, conservation, and community activities are fulfilled and/or to be fulfilled by using forest resources and how much of these can be replaced by using appropriate sustainable energy sources.



Design, build, and install appropriate energy technology in communities surrounding protected areas  Matching resources (waste, water, biomass, etc. to technology).  Knowledge exchange.  Technical specification.  Work with local communities and NGO partner, to implement new practices, assess their uptake, and measure the impact. 2. Determination of how the use of sustainable energy technologies can address land use issues:

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



Determine how access to energy improves livelihood and what impact does it have on the indigenous community‟s resilience to external pressures. Determine the factors that influence how energy consumption related economic activities result in these indigenous communities failing (or beginning to fail) to act as the guardian to protected areas, and become threats rather than benefits.

Furthermore, human development in the sector needs to take place (Figure 3). Academics and student mobility should be facilitated to build a strong local, sub-national, national, regional, and international network of existing and future practitioners and experts, based on on the ground hands-on understanding. The interdisciplinary teaching materials in new courses (and/or course components) through face-to-face lectures and online delivery (ODL); should allow for a bidirectional knowledge exchange in ecosystem conservation, ecosystem services and sustainable energy generation to cover policy, strategy, knowledge exchange and appropriate technology. 5. Extending the interface – from pilots to mainstreams Crucial to meet the principles of decentralization of renewable energy generation, multi level governance and local communities‟ engagement is key to successful sustainable energy appropriate technology implementation programmes. However, pre-determining the technology selection will run a high risk of failure. A range of appropriate renewable energy generation technologies have been proven to be successful through demonstration cases by inter alia the Agricultural Ministry, the Indonesian Institute of Sciences (LIPI), and the National Electricity Company (PLN), but there is as yet no identified approach to replicate and expand based on robust financial and business models. Discussions with the Centre for Appropriate Technology Development, Indonesian Institute of Sciences (LIPI-TTG) have highlighted the current needs for the rolling out of various appropriate technologies which have been tried and piloted in many locations for a nationwide community development. There clearly is a need to address key FEW issues from a holistic point of view (social, economic, political, and environmental). A joint social investment from the government and private sector is recognised as the primary component in identifying and bridging existing gaps in taking those successful pilot cases forward into a nationwide scalable and applicable implementation strategy. The joint social investment needs a welldefined and clear framework for project

management and financial accountability to avoid waste and inappropriate exploitation. Training for provincial governments to engage with appropriate technology projects is required, as is technical training through polytechnics to provide relevant local skills and to encourage new business development. This paper considers that there are 4 stages of activities required to move pilot projects in appropriate sustainable energy technology implementation into the mainstream as illustrated by Figure 4 and explained in detail in the subsequent sections. These stages should generate the significant knowledge needed to facilitate the provision of sustainable energy without jeopardizing ecosystem conservation requirements. These stages reflect the consideration that to be selfsustaining, any sustainable energy provision for remote, off-grid and/or subsistence communities must be based on utilising appropriate technology within a viable and local business case. The appropriate technology must be throughout the business model, i.e. the technologies for both generating and consuming energy must be appropriate for the community using it. This is so that the community can build, repair, manage, and directly benefit from its own energy systems. Conversely, there needs to be recognition that, at some point, increased economic activity in areas with sensitive ecosystems will increase the community‟s impact on their surrounding ecosystems beyond some acceptable level; but what is an acceptable level and who should determine it? 5.1 Stage 1 The first stage is the communication phase. It is important to properly extract the appropriate information reflecting the actual needs of the local people, since the optimal use of any technology depends entirely on its interface with the people who use it. Thus, the early positive involvement at local level is vital to the success of village development. The villagers possess the knowledge and the pertinent skills to ensure full and productive engagement with the proposed technology. 5.2 Stage 2 The second stage is the construction of a closed loop implementation of the solutions for the holistic management of the issues relating to FEW nexus. Such approach should optimise production whilst minimising waste. The construction of the closed loop approach involves two phases: 1) capacity building and creation of human capital development (teaching/training) modules, and 2) development of a set of recommendations for National, and Local Government institutions.

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The first step is a process that should be initiated by first engaging with appropriate educational institutions, using participatory methodology which involves local communities, iterating teaching/training modules development between user (local community) and provider (local vocational/educational institutions), and developing paper based and online material for knowledge exchange and information dissemination. This step should not only prepare local people and institution

throughout the implementation of the closed loop solutions, but also generate knowledge pertinent to the second step of providing recommendations for the government. The second step generates specific sets of outcomes. It should recommend policy formulation, intervention strategy for replication, and strategy for access to financing mechanisms. This step should also identify opportunities for sustainable integration of F-E-W within local businesses.

Figure 4. Road Map - from Pilot to Mainstream 5.3 Stage 3 This is the process aiming to generate feedbacks towards the results from Stage 2. Stage 3 would primarily rely on local data gathering which should be undertaken to ensure that existing indigenous wisdom (both from rural/coastal communities) is fully utilised and respected. The data gathering would mainly consist of the consideration and collation of existing best practices, pilot projects, indigenous wisdom, and community initiatives across Indonesia. Several essential steps are required to ensure that pertinent data are collected, such as:   

Creation of database of lessons learnt (both positive and negative) Quantitative and qualitative data analyses on the issues identified in previous sections Regularly updated monitoring and evaluation recording of pre, during, and post project situations



    

Initiating community engagement and assessment of the community‟s existing developmental position, their needs, aspirations and resources Support for community enterprise engagement and access to finance Assessing the local meteorological, marine, hydro, bio-fuel and waste resources available to the village Assessing the existing economic, domestic and commercial activities, identify existing skills availability Assessing existing energy consumption magnitude and daily patterns Identifying the site-dependent appropriate technologies for energy generation and energy consumption as part of embedded economic activities

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5.4 Stage 4 In conjunction with Stage 3, this stage also attempts to seek feedback from the local communities. However, the collected feedback would be primarily more related with how a visible demonstration incorporating integrated appropriate technology is to be developed based on the SOP‟s created, incorporating the views and needs of local actors and stakeholders. The demonstration would primarily take the form of a model village site. The model village site would be selected from a list of options that would be created based on the representativeness of Indonesian villages (rural and/or coastal). The selection itself would consider 1) the variety of potential issues in FEW confronted by local communities and the feasibility of such location in accommodating a range of appropriate energy technology systems as demonstrable means for problem solving, 2) existing stakeholders interest where local communities can be engaged to be part of the problem solving exercise, where support from local government is committed, and buy ins from businesses visible, 3) accessibility of site to researchers involved, expertise and other potential supplier and user of the model village services. 6. Conclusion Indonesia has thousands of small communities depending on small-scale activities for their livelihood e.g. fishing, agriculture, aquaculture and forestry. These are often significant ecosystem and natural resource challenges where natural resources management and economic development create conflicting pressures. On the other hand, with its high number of population, Indonesia needs to fulfill the ever increasing food and clean water demands. Meanwhile, the primary requirement for socioeconomic development in any society is access to energy resources, not only electricity, which can enhance existing activities and create new initiatives. To fully accommodate the socio-economic developments while acknowledging the needs of ecosystem and natural resources management, the holistic approach of food-energy-water (FEW) nexus should be adopted in both strategies. The holistic view of FEW nexus is a strategy that considers the balancing act between the four “P”s of sustainable practice (i.e. Politics, People (social aspect), Planet (environmental aspect), and Profit (Economic aspect). The ackowledgment of the role of these four “P”s in every elements of food, water and energy enables the local knowledge and feedback throughout the whole decentralised interface of the system. This will benefit in the prevention of cyclic friction between food and energy generation processes and the needs of

protected areas. The local knowledge gathered and consulted throughout the holistic FEW nexus framework should ensure that the method used should be self-sustaining and does not conflict with ecological need of the areas. References [1] Anderson, G. Q., Fergusson, M. J. (2006), “Energy from Biomass in the UK: Sources, Processes and Biodiversity Implications”, Ibis, 148 (s1), 180-183. [2] Barnes, A. D., Jochum, M., Mumme, S., et al. (2014) “Consequences of Tropical Land for Multitrophic Biodiversity and Ecosystem Fuctioning”. Nature Communication, 5: 5351 doi: 10.1038/ncomms6351. [3] Butchart S. H., Walpole M., Collen B., et al. (2010), “Global Biodiversity: Indicators of Recent Declines”, Science, 328 (5982), 1164 – 1168. [4] Calvert, K., Mabee, W. (2015). “More Solar Farms or More Bioenergy Crops? Mapping and Assessing Potential Land-Use Conflicts Among Renewable Energy Technologies in Eastern Ontario, Canada”. Applied Geography, 56, 209-221. [5] Carlson K. M., Curran, L. M., Asner G. P., et al. (2013) “Carbon Emission from Forrest Conversion by Kalimantan Oil Palm Plantations”. Nature Climate Change, 3, 283287. [6] Chicco, G., Mancarella, P. (2009). “Distributed Multi-Generation: A Comprehensive View”. Renewable and Sustainable Energy Reviews, 13 (3), 535-551. [7] Conservation International (2014), Indonesia [Online], Available from: http://www.conservation.org/where/Pages/Indo nesia.aspx (Accessed 20 May 2015). [8] Danielsen, F., Beukema, H., Burgess, N.D., et al. (2009) “Biofuel Plantations on Forested Lands: Double Jeopardy for Biodiversity and Climate”. Conservation Biology, 23, 348-358. [9] Department for International Development (2013). Operation Plan 2011 – 2015 UK Climate Change Unit Indonesia (UKCCU) [Online].Availablefrom: https://www.gov.uk/government/uploads/syste m/uploads/attachment_data/file/208924/Indone sia.pdf [Accessed 18 November 2013]. [10] Eghenter, C. (2000), Mapping Peoples‟ Forests: The Role of Mapping in Planning Communitybased Management of Conservation Areas in Indonesia, Washington DC: Biodiversity Support Program.

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[11] Food and Agriculture Organisation of the United Nation (FAO) (2005), Global Forest Resources Assessment 2005: Progress Towards Sustainable Forest Management, Rome: FAO Forestry Paper 147. [12] Groom, M. J., Gray, E. M., Townsend, P. A. (2008). “Biofuels and Biodiversity: Principles for Creating Better Policies for Biofuel Production”, Conservation biology, 22(3), 602609. [13] Heinrich J., Heine, S. J., Norenzayan A. (2010). “The Weirdest People in the World”. Behavioural and Brain Sciences 33: pp. 61– 135 doi: 10.1017/S0140525X0999152X [Online]. Available from: http://hci.ucsd.edu/102b/readings/WeirdestPeop le.pdf (Accessed 5 September 2014) [14] Inger, R, Martin J. A., Stuart B., et al. (2009), "Marine Renewable Energy: Potential Benefits to Biodiversity? An Urgent Call for Research", Journal of Applied Ecology 46 (6), 1145-1153. [15] IRRI (2015), “Rice is Key to Indonesia‟s Food Security, Officials Say” [Online], Available from: http://irri.org/news/mediareleases/rice-iskey-to-indonesia-s-food-security-officials-say (Accessed 24 May 2015). [16] Karger, C. R., Hennings, W. (2009). “Sustainability Evaluation of Decentralized Electricity Generation”. Renewable and Sustainable Energy Reviews, 13 (3), 583-593. [17] Koh, L.P., Wilcove, D.S. (2008) “Is Oil Palm Agriculture Really Destroying Tropical Biodiversity?”. Conservation Letters, 1, 6064. [18] Koh, L.P., Miettinen, J., Liew, S.C., Ghazoul, J. (2011) “Remotely Sensed Evidence of Tropical Peatland Conversion to Oil Palm”. Proceedings of the National Academy of Sciences of the United States of America, 108, 5127-5132. [19] KPMG International, 2012. Expect the Unexpected: Building business value in a changing world. KPMG International [20] Kruisen, J.H.J., Owen, A., Turner, N., Garniati, L., (2012). The Fourth Dimension of Sustainable Practice. http://cesun2012.tudelft.nl/images/d/d0/Kruijse n.pdf [accessed 12 July 2013]. [21] Land Use Consultant and TV Energy (2010). Review of Renewable and Decentralised Energy Potential in South East England [Online]. Available from: http://www.climatesoutheast.org.uk/images/upl oads/Renewables_Potential_in_SE.pdf (Accessed 27 May 2015).

[22] Marquardt, J. (2014), „A struggle of Multi-level Governance: Promoting Renewable Energy in Indonesia‟, Energy Procedia, 58, 87-94. [23] McLellan, B., Florin, N., Giurco, D., et al. (2015). “Decentralised Energy Futures: The Changing Emissions Reduction Landscape”. Procedia CIRP, 29, 138-143. [24] Ministry of National Development Planning/National Development Planning Agency (MP3EI) (2012), Masterplan for Acceleration and Expansion of Indonesia Economic Development 2011 – 2025, Jakarta: Coordinating Ministry for Economic Affairs 2011. [25] Naylor, R.L., Liska, A.J., Burke, M.B., et al. (2007) “The Ripple Effect: Biofuels, Food Security and the Environment”. Environment, 49, 30-43. [26] Oglethorpe, J., Ericson, J., Bilsborrow, R. E., Edmond, J. (2007), People on the Move: Reducing the Impact of Human Migration on Natural Ecosystem, Washington DC: World Wildlife Fund and Conservation International Foundation. [27] Omer, A. M. (2008), “Energy, Environment and Sustainable Development”, Renewable and Sustainable Energy Reviews, 12 (9), 22652300. [28] Pelc, R., Fujita, R. M. (2002), “Renewable Energy from the Ocean”, Marine Policy, 26 (6), 471-479. [29] Pepermans, G., Driesen, J., Haeseldonckx, D., et al. (2005). “Distributed Generation: Definition, Benefits and Issues”. Energy policy, 33 (6), 787-798. [30] Practical Action. Decentralised Energy Scheme [Online]. Available from: http://practicalaction.org/decentralisedenergy (Accessed 27 May 2015). [31] Quanz, C., Bunbury, N., Fleischer-Dogley, F. (2013), “Improving the Sustainable Operation of a World Heritage Site: Increasing Energy Efficiency and Implementing a Renewable Energy System on Aldabra Atoll, Seychelles”. Parks, 19.2, 47–58. [32] Ramchurn, S. D., Vytelingum, P., Rogers, A., Jennings, N. R. (2012). “Putting the 'Smarts' Into the Smart Grid: A Grand Challenge for Artificial Intelligence”.Communications of the ACM, 55 (4), 86-97. [33] Republika (2014), “Jokowi to Allocate Larger Budget for Research” (Online), Available from: http://www.republika.co.id/berita/en/nationalpolitics/14/09/16/nbzvjm-jokowi-to-allocate-

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larger-budget-for-research (Accessed 24 May 2015). [34] Rosen, M.A., 2009. Rosen, Energy sustainability: a pragmatic approach and illustrations, Sustainability. pp. 55-80. [35] Saha, P.C., 2003. Sustainable energy development: a challenge for Asia and the Pacific region in the 21st century. Energy Policy, 31, pp. 1051-1059. [36] South Centre (2008). The Role of Decentralized Renewable Energy Technologies in Adaptation to Climate Change in Developing Countries. Analytical Note SC/GGDP/AN/ENV/5 [Online]. Available from: http://www.southcentre.int/wpcontent/uploads/ 2013/07/AN_ENV5_Role-Of-DecentralizedRenewable-Energy_EN.pdf (Accessed 27 May 2015). [37] Tolisano, J., Vosper, A., Hatchwell, M., Victurine, R. “The Ape in the Machinery: A Status Report on Great Ape Conservation in Natural Resource Extraction Zones in Central Africa” [Online], Arcus Foundation, Available from: http://www.stateoftheapes.com/wpcontent/uploads/2014/07/The-Ape-in-theMachinery.pdf (Accessed 18 May 2015). [38] The Jakarta Post (2011), “Mining, Plantation Disputes to Intensify” [Online]. Available from: http://www.thejakartapost.com/news/2011/12/2 9/mining-plantation-disputes-intensify.html (Accessed 05 June 2015).

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PRELIMINARY STUDY OF AGRIBUSINESS DEVELOPMENT BASED ON ALOE VERA (Case Study in Micro, Small and Medium Enterprises: Sari Kumetap Subang) 1

N. Rahman, 2W. Agustina, 3R. I. Tribowo, 4C. E .W. Anggara, 5R. C. Erwan and 6A. Wulansari 1,2,3,4,5

Center of Appropriate Technology Development - Indonesian Institute of Sciences K.S. Tubun No.5, Subang, West Java 41213. Telp. 0260-411478 Fax. 0260-411239 Email: [email protected] 6

Research Center of Biotechnology - Indonesian Institute of Sciences Cibinong Science Center, Jl Raya Bogor Km 46, Cibinong-Bogor , West Java 16911 Telp. 021- 8754587 Fax. 021- 8754588

Abstract : Aloe vera not merely can be sold in the form of plants, but also can be processed and produced become kinds of food, beverage, herb medicine and cosmetic. Variety of Aloe vera that being cultivated is Barbadensis which contain substance for human body need, like vitamin of A, B1, B2, B6, B12, E and C. These plants purportedly can also heal for example diabetes and heart diseases. Processed product based on Aloe vera that developed by UKM (Micro Small Business) Sari Kumetap relatively so many, like aloe essence, aloe tea, aloe ice, aloe chicken porridge, mixture of aloe herb with multifarious rimpang herbs, cosmetic like lulur (skin enlightened) and moisturizer. Nevertheless business activity that conducted by the UKM still must improve its quality and also production quantity with applying some technologies including sufficient equipments and packaging technique which conducted through Iptekda (Technology implementation for targeted area) program. From existing various Aloe vera product, through this Iptekda activity will be more emphasized at 3 prior products that is Aloegin, Aloe Tea and Aloe ice. Business of aloegin production with production capacities as high as 50 litres aloegin can produce 100 bottles @ 500 ml aloegin with price of IDR. 60,000.-/bottle will get IDR. 3,015,000.- gross profit for every process/month, so this business is quite feasible to be developed. The production capacities of Aloe vera processing, in this case tea and aloe ice, are also still follow market absorption. Utilization of tissue culture technology which produce 500 bottles of plants seed (each bottle content 5 seed of Aloe vera plant) with its price as high as IDR. 10,000/bottle will get IDR. 2,354,000,- gross profit for every process/month, so this business is quite feasible to be developed. The equipments that will be introduced through Iptekda – LIPI (Indonesian Institute of Sciences) activity to complete processing equipments and tissue culture of Aloe vera i.e. Laminar flow, Cold storage, Greenhouse, Gas stove, Oven, Mixer, Shaker, Blender, Sealer, Digital Refrakto Meter, pH metre and Digital weighing-machine. The UKM Sari Kumetap is led to become center of agroindustry business development based on Aloe vera material that runs commercially (profitable), expand and sustained. Keyword: Agribusiness, Aloe vera, UKM Sari Kumetap, Tissue Culture 1. INTRODUCTION 1.1 Backgrounds Aloe vera not merely can be sold in the form of plants, but also can be processed and produced become kinds of food, beverage, herb medicine and cosmetic. Selling price in the form of plants is relatively high, and does so if processed in the form of food, beverage of herb medicine and cosmetic. The type of Aloe vera that being cultivated is Barbadensis type that contain substance for human body need, like vitamin of A, B1, B2, B6, B12, E and C. This plants purportedly can also heal for example diabetes and heart diseases [1][3]. Aloe vera has strategic role in order to improve source of society earnings and labour absorption. Processed product based on Aloe vera that developed by UKM (Micro Small Business) Sari Kumetap relatively so many, like aloe essence, aloe

tea, aloe ice, aloe chicken porridge, mixture of aloe herb with multifarious rimpang herbs, cosmetic like lulur (skin enlightened) and moisturizer. Nevertheless business activity that conducted by the UKM, it can be told that still relatively simple, especially in the case of utilizing of technology like equipments and packaging. Its limitation causes low productivity, meanwhile according to Herwanto (40 years old), the owner of UKM Sari Kumetap, the level of demand is quite high. In order of effort to improve quality and quantity of Aloe vera processing production, then it is required an application of appropriate technology in kind of technology aid like equipments technology and processing process. In order to support the production effort, the UKM besides cultivate its Aloe vera by it self by using the existing empty farms like plantation area and yard. In the other hand

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the UKM also empower the society/farmer to plant Aloe vera and even plan will more amount of offer to farmer to plant Aloe vera in the future. The development of Aloe vera agriculture cultivation to farmer need seed that quite a lot, for that in the seed availability will be done by application of household scale of tissue culture technology. The application of processing technology and equipments and tissue culture is expected can improve quality and productivity of production effort of processed product bases on Aloe vera that has been developed since a long time ago by UKM Sari Kumetap, in doing so can develop production for local, national even international need. 1.2 Aim The preliminary study is conducted to get base that rely on data that obtained in objective in activity location, in this case at Micro Small Business (UKM) Sari Kumetap, in doing so at step of development activity of agribusiness based on Aloe vera in the UKM will fulfill the target. 1.3 Methodology The method which is used in the elementary study is to conduct direct survey to the location of UKM (Micro Small Business), in this case Micro Small Business Sari Kumetap. Included in it is problems discussion (a kind of focus group discussion) between team of Pusbang TTG and the organizer of UKM. The problems which are discussed e.g. profile of the UKM, cultivating technology and the equipment, raw material and marketing, management of bookkeeping and any that required for the development of agribusiness based on Aloe vera. Analysis of SWOT (Strengths Weaknesses Opportunities Threats) is conducted especially for looking the positive factors that must be defended, built even improved. Looking for the weakness that must be improved altered or desisted. The Opportunity must be prioritized, caught, built and

Figure 1.

Aloegin

Figure 2.

Aloe Tea

optimized. The threat must be overcome or minimized and managed [6]. 1.4 Activity Location The activity of the elementary study is conducted in the UKM Sari Kumetap that the address is at Cagak street - View Garden Residence - Block D24 No.11 Palasari village, sub-district of Ciater, district of Subang, province of West Java by distance around 20 km from Subang city toward Bandung. 2. RESULT AND DISCUSSION 2.1 Business Profile of the UKM Sari Kumetap The UKM Sari Kumetap has conducted a business in sector of Aloe vera cultivation and it‟s processed since 2010 that pioneered by Herwanto (40 years) and friends. In this Aloe vera cultivation business, the UKM besides offering the advantage from harvesting result of Aloe vera also offering the advantage through purchasing weeds (herb) and then processed or utilized also for business of food production such as flaky pegagan and flaky sintrong. Processing from Aloe vera that already and is being conducted e.g.: yogurt, cocktail, juice, jam, dodol, syrup, bandrek, tea. Besides food and beverage, product from Aloe vera is used also for cosmetic materials like cleanser, scrub, masker and liquid soap. 2.2 Technology of Production Process One of horticulture product that gets priority in production development is Aloe vera . This plants is recognized easy grow and cultivated in Indonesia. This plants actually is not genuine Indonesia plants but indigenous from Africa [8]. From Aloe vera plants can be produced a variety of forms and wide varieties of product type for example beverage product, tea, cosmetic, and many processed food. Processing aim is one of other things to lengthen the durability or conserves. In the other hand processing can make more useful varieties product and more has economic value.

Figure 3.

Aloe Ice

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Processing from Aloe vera that already and is being conducted for example: yogurt, cocktail, juice, jam, dodol, syrup, bandrek, tea. Besides product food and beverage from aloe vera, it is used also for cosmetic materials like: cleanser, scrub, masker, liquid soap. From the existing varieties of product, through the activity of Iptekda-LIPI will be more emphasized at 3 prior products i.e. Aloegin (Figure 1), Aloe Tea (Figure 2) and Aloe Ice (Figure 3). The processing flowchart of the three of product can be seen at Figure 4, Figure 5, and Figure 6.

Figure 4. Flowchart of ALOEGIN Production Process

2.3 Technology of Tissue Culture at Household Scale The main principle of tissue culture technique is the cultivation of plant tissues and organs under aseptic conditions in controlled environments. In general tissue culture is conducted at laboratory scale. By simplification of the equipments, the materials which are used and the optimum environment, this technique can be conducted in household scale. The cultivation of Aloe vera in district of Subang is rarely done. One of the reasons is limited plants seed that generally from shoots/saplings. Application of tissue culture technology is expected to be a solution to overcome that problem [5]. Tissue culture technique or in vitro technique can produce uniformed plant, similar to the parent plant. It produced many plants in a short time, it takes at least one week for first shoot growth compared with conventional propagation which takes more than one month. MS medium fortified with 1 mg/l BAP was the optimum medium for shoot multiplication (Figure 7). In 10 weeks, single in vitro shoot can multiply to 10-15 shoots [2]. This technique does not require a large place and does not depend on season so it can be done throughout the year [3].

Figure 5. Flowchart of Aloe Tea Production Process

Figure 6. Flowchart of Aloe Ice Production Process

Figure 7. The growth of Aloe vera plant from in vitro shoot through acclimatization

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Introduction of household scale tissue culture technique to UKM Sari Kumetap is expected could increase their knowledge, experience and most importantly could increase their revenue. From benefit cost projection, utilization of tissue culture technology for Aloe vera plants, which produce 500 bottles of plants seeds (each bottle content 5 seeds of Aloe vera plants) with its price as high as IDR. 10,000//bottle will get IDR. 2,354,000,- gross profit for every process/month, so this business is quite feasible to be developed. 2.4 Production Capacity and Benefit Cost Projection

The production capacity of Aloe vera processing, in this case aloegin still follows market absorption. At this time the market absorption is in the average of 100 bottles for the size of 500 ml per month. For 500 ml aloegin required 2 kg of raw material of Aloe vera leafs. Benefit cost projection can be seen at tables 1. From table 1 indicate that business of aloegin production with production capacity of 50 litre aloegin can produce 100 bottles @ 500 ml with price of IDR. 60,000.-/bottle, will get IDR. 3,015,000.gross profit for every processes / month, so this business is quite feasible to be developed.

Table 1. Benefit cost projection of aloegin product

The production capacity of Aloe vera processing, in this case tea and aloe ice are also still follow market absorption. At this time according to explanation from Herwanto (owner of the UKM) its sale is relatively still profitable and quite feasible to be developed. 2.5 The Technologies that Will Be Introduced The operational of technical activity that will be conducted is technological transfer/experience transfer to the UKM that had been owned by LIPI currently, including from household scale of tissue culture pilot plant. Substantially the existence of the Center of Appropriate Technology Development Indonesian Institute of Sciences (Pusbang TTG – LIPI) becomes reference center in development and application of appropriate technology for the society (UKM) and provides appropriate technology need for the society (UKM). In accordance with the existing experience, the technologies that will be introduced to the UKM Sari Kumetap are:  Technology of raw material and support materials procurement

 Technology of processing through operation of machine equipments and Aloegin, tea and ice production engineering.  Technology of household scale of tissue culture.  Technology of product quality control, labeling, packaging, marketing, promotion, record-keeping management, bookkeeping. The equipments that will be introduced through the activity of Iptekda – LIPI to complete the equipments of processing and tissue culture of Aloe vera for example : Laminar Flow, Cooler Cupboard, Greenhouse, Gas Burner, Oven, Mixer, Shaker, Blender, Sealer, Digital Refracto Meter, pH metre and Digital Weighing Machine. In running and developing the UKM Sari Kumetap are required business management that cover process management (raw material, equipments, and production process), financial management (recordkeeping and transaction bookkeeping, job contracts and cash flow) and marketing management. For that the training activity to support the UKM Sari Kumetap business is conducted along with the running Iptekda activity.

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2.6 Technological Transfer Fund Managements The technological transfer fund that used for management of the UKM will be realized in the form of required equipments, raw material and packaging materials that match with the UKM need. The technological transfer fund hereinafter will be managed by Koperasi. Koperasi has function as a place of provider/levying of equipments , materials of the UKM need, development mediator of the UKM, executor of field activity, owner and organizer Iptekda's fund and management assistance of the UKM. As for the profit sharing of the activity result is agreed on that profit sharing is allotted as follows: 60 % of net profit for the UKM Sari Kumetap and 40 % for Koperasi. The fund that come into Koperasi will be used for working capital addition and develop the UKM Sari Kumetap it self or other UKM on a sustainable way. The fund management scheme of the technology transfer can be seen at Figure 8.

will facilitate it coming into the export market. Give aid by the shape of technology or equipments to run its business. Marketing access can be conducted through exhibition and virtually exhibition/internet. Thus the UKM products posted or vended at many web sites, its aim is in order to broader the range of marketing access, included cover export scale [6]. Like has been mentioned at introduction part, where the market potency of the product of food processed like Aloegin and Tea are quite promising. Whereas for product of aloe ice is targeted to schools and small shop near by the UKM. As for marketing strategy that taken by the UKM Sari Kumetap, that at this time with Aloegin as the prior product, is directly selling to the consumer that already become its customer in Subang and its surroundings. 2.8 Analysis of SWOT (Strengths Weaknesses Opportunities Threats) The Sari Kumetap MSME’s/UKM Analysis of SWOT is an identification of various of factors systematically to formulate pattern of business development strategy the UKM Sari Kumetap. This analysis is relied on logic to develop strength, overcome weakness, utilize opportunity and overcome or minimize the threat [6][7]. The strength in business development in the UKM Sari Kumetap for example: raw material (Aloe vera) available along the season, available enough the human resources, the required technology is the technology that is not sophisticated, deep support from stake holder by promote the local prior product, product of natural Aloe vera by utilizing local resources result.

Figure 8. Fund management scheme of the technology transfer 2.7 Market Potential and Way to Market The Product This moment there are 5 main problems that become constraint that faced by the UKM community, for example access marketing, capital, management, technology and partnership access. To overcome it, UKM requires support from many parties. Besides the local government, the central government is expected has important role in develop the UKM through small and middle trade center (PDKM) Commerce Department tries to solve the problem as mentioned above [6]. One of way that conducted is by exhibition of the UKM products either in home country or at foreign country. Build up partnership with modern markets. By draw the UKM products to these modern markets

The weakness in business development in the UKM Sari Kumetap: production capacity and capital owned are limited, technical and managerial ability are still limited, market range still rely on local market. The opportunity in business development in the UKM Sari Kumetap: the siding of the stake holder to the UKM Sari Kumetap is high enough, the society need of Aloe vera product is growing up, the business location is relatively close to tourism place (Ciater, Lembang, Tangkuban Parahu, product marketing network is very opened). The threat in business development in the UKM Sari Kumetap : competition with big company that has been recognized, the pineapple plantation of society property tends to become people settlement, electricity and fuel tariff increased, ASEAN Economic Community (AEC) 2015 [6]. Based on the analysis result of SWOT to the UKM Sari Kumetap, then the alternatives strategy pattern that can be developed for example:

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Strategy to maximize the internal strength to utilize external opportunity that is: extend marketing network by recruiting experienced persons, maximizing sale by multiply customer. Thus this strategy is conducted by maximizing existing strength for getting advantage, getting market segment to reach sustained business development. Strategy to overcome the internal weakness to utilize opportunity external is conducted by improving the promotion/advertisement, apply managerial system assistance especially in the field of standard recordkeeping and bookkeeping. Strategy to maximize the strength and effort to overcome or minimize the threat is conducted by improving innovation, product competitive ability and product quantity, product certification and waste management. Strategy to overcome the internal weakness for minimizing external threat are conducted by care of productivity, hygienist and efficiency of production cost and increasing the skill and prosperity SDM [6]. 3. CONCLUSION and SUGGESTION The type of Aloe vera that being cultivated is Barbadensis type that contain substance for human body need, like vitamin of A, B1, B2, B6, B12, E and C. This plant purportedly can also heal for example diabetes and heart diseases. Aloe vera not merely can be sold in the form of plants, but also can be processed and produced become kinds of food, beverage, herb medicine and cosmetic. From the existing varieties of product, through the activity of Iptekda-LIPI will be more emphasized at 3 prior products i.e. Aloegin, Aloe Tea and Aloe Ice. Business of aloegin production with production capacity of 50 litre aloegin can produce 100 bottles @ 500 ml with price of IDR. 60,000.-/bottle, will get IDR. 3,015,000.- gross profit for every processes / month, so this business is quite feasible to be developed. The production capacity of Aloe vera processing, in this case tea and aloe ice are also still follow market absorption. Utilization of tissue culture technology which produce 500 bottle of plants seeds (each bottle content 5 seeds of Aloe vera plants) with its price as high as IDR. 10,000//bottle will get IDR. 2,354,000,gross profit for every process/month, so this business is quite feasible to be developed. The equipments that will be introduced through the activity of Iptekda – LIPI to complete the equipments of processing and tissue culture of Aloe vera for example : Laminar Flow, Cooler Cupboard, Greenhouse, Gas Burner, Oven, Mixer, Shaker, Blender, Sealer, Digital Refracto Meter, pH metre and Digital Weighing Machine. The UKM Sari Kumetap is led to become center of agroindustry business development based on Aloe vera material

that runs commercially (profitable), expand and sustained. Based on the analysis result of SWOT to the UKM Sari Kumetap, then the alternatives strategy pattern that can be developed for example: Strategy to maximize the internal strength to utilize external opportunity that is: extend marketing network by recruiting experienced persons, maximizing sale by multiply customer. Thus this strategy is conducted by maximizing existing strength for getting advantage, getting market segment to reach sustained business development. Strategy to overcome the internal weakness to utilize opportunity external is conducted by improving the promotion/advertisement, apply managerial system assistance especially in the field of standard recordkeeping and bookkeeping. Strategy to maximize the strength and effort to overcome or minimize the threat is conducted by improving innovation, product competitive ability and product quantity, product certification and waste management. Strategy to overcome the internal weakness for minimizing external threat are conducted by care of productivity, hygienist and efficiency of production cost and increasing the skill and prosperity. 4. ACKNOWLEDGMENT Thank you we say to The Head and Staff of Center of Appropriate Technology Development Indonesian Institute of Sciences (B2PTTG-LIPI) in Subang and UKM Sari Kumetap, especially to: Drs. Sukirno MS and Herwanto, Local and related Institution to the all aids that have been given. REFERENCES [1] Badan Pusat Statistik Kabupaten Subang, 2010, Subang dalam angka, Subang. [2] Imelda M, Wulansari A, Sari L & Erlyandari F. 2006. Peningkatan kadar aloin lidah buaya melalui embriogenesis dan mutagenesis. Laporan Teknik Kegiatan Penelitian Pusat Penelitian Bioteknologi, DIPA tahun 2005 : 3240. [3] Rahman, N., dkk., 2014, Pengembangan Usaha Agribisnis Berbasis Lidah Buaya di Usaha Kecil Mikro Sari Kumetap di Kabupaten Subang, Proposal kegiatan penerapan ilmu pengetahuan dan Teknologi di daerah (iptekda) lipi Tah\un 2015. Subang. [4] Rahman, N. dan Rahayuningtyas, A., 2013, Penerapan Teknik Kultur Jaringan Dalam Rangka Penyediaan Bibit Singkong Jenis Darul Hidayah Dalam Upaya Peningkatan Mutu Produk Olahan Singkong, Prosiding Seminar Nasional dan Workshop Peningkatan Pemanfaatan Inovasi dalam Menanggulangi

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Kemiskinan, Bandung, Oktober 2013.

30 September- 1

[5] Sriyanti dan Daisy, P., 1994, Teknik Kultur Jaringan, Kanisius, Yogyakarta. [6] Sukirno, 2014, Pattern of Appropriate Technology Development of Processing of Pineapple Fruit in Together Business Group of Alam Sari Subang, Draft makalah seminar, Pusbang TTG-LIPI, Subang, 12 pp. [7] http://www.academia.edu/5090849/Pengertian_ analisis_SWOT,17 Februari 2015. [8] http://www.mellindapsari.worldpress.com, Juli 2014.

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Potential Development of Arengapinnata, Merr Based On Local Knowledge (Case Study of Rejang Lebong) Eki Karsani Apriliyadi a), Diki Nanang Surahman a), Hendarwin M Astro a)* a)

Center of Appropriate Technology Development, Indonesian Institute Of Sciences (IIS), Jl. K.S. Tubun No. 5 Subang 41213, Indonesia Tel.: +62-260-411478; fax: +62-260-411239. E-mail address : [email protected]

Abstract: Arengapinnata, MERR is potential commodity for handling food suffering and it easy to adaptation in every agroclimate, start from low land until high land 1400 metres above sea level (Effendi, 2009; Ditjen Perkebunan, 2004). Arengapinnata, MERR has become of the main income for Rejang Lebong Perfecture people. Recently, the local farmers making brown sugar from sweet sap of Arengapinnata, MERR, and they has been cultivate in their own land. The local knowledge of brown sugar production has been made by generation to generation and its become routine activity. This main activity hasbecame of development program collabarted between Center of Appropriate Technology Development and the government of RejangLebong Prefecture. The goals of this program are giving added value and income generation for local farmers. To increase the potency of Arengapinnata, MERR by using appropriate technology as a vehicle for optimizing local resources. Keywords : Arengapinnata, MERR; local knowledge; RejangLebong; appropriate technology 1. Introduction Biodiversity in Indonesia has strong related with the local people i.e. Arengapinnata, MERR. Almost through out Indonesia can be found this commodity. The plantation of Arengapinnata, MERR has potency to develop. The main product of Arengapinnata, MERR is sweet sap. Its processing into various of products such as brown sugar, beverage, vinegar, and liquor. In the other hand, part of this plant can be used for producing food material. Mostly the products are producing by local farmers not industrial scale. This situation appeared because the intensive cultiovation not found yet. Rumokoi (2004) explains, from data processing which bring out by Ditjenhutbun in 2013, growth rate of plantation area in several provinces are about 60.482 ha with production capacity of brown sugar about 30.376 tons/year. The largest area and production of brown sugar can be found in West Java Province 13.135 ha with brown sugar production 6.686 tons/year, Papua 10.000 ha with production of brown sugar 2.000 tons/year, South Sulawesi 7.293 ha with production 3.174 tons/year, and North Sulawesi 6.000 ha with production of brow sugar 3.000 tons/year. In Sumatera (Bengkulu province), this comodity can be found in every prefectures with total area about 2278 ha and the production in 2011 is about 2471 (Dinas Perkebunan Bengkulu). The area of aren cultivation in every prefectures are Bengkulu Selatan 91 Ha, Bengkulu Utara 29 Ha, Kepahiyang 163 Ha, Lebong 254 Ha, Mukomuko 53 Ha, Rejang Lebong 2292 Ha, and Seluma 46 Ha.

The Arengapinnata plant has an adaptation capability in many soil condition, agroclimate, and high tolerance to multiple cultivation with others comodities include woddy plants, growing fast and has large root area and wide canopy also. Its suitable to develop and solve the problems in marginal soil. From the aspect of utility, aren plant has various function because the whole parts of aren plant can be use for many kind of products, for examples sweet sap can be used for brown sugar and liquor. The sweet sap can be used for material of producing bioethanol, and its known as alternative material for bio-fuel (Widyawati 2011). The seed of aren plant fruits can be used for food material, one of the food that can be produced from aren plant seed are cocktail and sweet pickles. The strach flour is produced from the log of aren plant, this starch can be processed into starch noodle. ”Ijuk” that sticking on the aren plant can be used for roof material, filtering water and etc. ”Lidi” that produced from the aren leaves can be used as handy craft material, broom and etc. Young leaves of aren plant can be used for ciggaretes material, natural packaging and etc. The roots has function for soil particle bounding and it used for minimazing erotion and landslide (Widyawati 2011). In the other hand, aren plant also can produce biomass both above or under the soil, and it has major function for CO2 cycle (Syakir dan Effendi, 2010). Some province in Indonesia have potency of aren plant. The provinces are West Java, Bengkulu, Papua, Sulawesi, etc. Mostly the usage of aren plant still limited and using mannualy processing.The processing itself, farmers using traditional or local knowledge. Each of province has differents type of

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local knowledge for processing aren plant. It depends on environtment condition and spreading process technic of processing from the other area in Indonesia. Rejang Lebong Perfecture is the one of area in Indonesia that have abundant resources to develop. The usage of aren plant has already done, but still they using tradionally processes.To consider of potency resources or comodity of aren plant in Rejang Lebong Prefecture, it need efforts to developing the usages of aren plant based on local knowledge. The program itself hopefully can increase diversification of food productsfrom aren plant and also can improve the quality of the products at the end it can support income generation of the farmers.

4. The Overview

2. Problems

Aren become one of abundant potential natural resources, and most have been harnessed into palm sugar. At the present time a lot of people who've been able to cultivate the palm trees in the gardens they have. Previously they only rely on palm trees that grow naturally in the forest. Farming activities carried out in an effort to develop the business of making palm sugar where current demand is increasing. Marketing is up to the various regions in Sumatra, especially from West Sumatra where this palm sugar is raw material for making soy sauce.

In processing of brown sugar from aren, the problems has been identified as below : 1) The quality of products from aren sweet sap which is brown sugar still has low quality; 2) The level of knowledge and skill to diversification products from aren sweet sap is low. 3. Method The type of this research was applied and field activity. The method was used action reasearch that has purposes to capture and giving the solve to the problems that has been identified. The research itself is also involving the local farmers as the object of research for to find the problem solving in their producing activity. FocusGroup Discussion (FGD) was used for digging and collecting information that related with the problems of their activity. In this research, there were 20 persons as respondens which are from farmers and public servants of the goverment in Rejang Lebong Perfecture. Depth Interview was used for collecting main and important informations related to posibillities for bussiness development in small and medium enterprises. The approach of this research is by using technical assistance. Technical Assistance might be broadly defined as the provision of programs, activities, and services to strengthen the capacity of recipients to improve their performance with respect to an inherent or assigned function (Wright, 1978).The technical assistancethat was applied in trainning format in order to result increasing and improving their skill and knowledge by using local knowledge. At the end the products that their produced from aren plant has good quality and can be marketed through out Rejang Lebong Perfecture.

Rejang Lebong is the one of perfecture in Bengkulu Province that has various potential natural resourcesand it can support agroindustry development. Some of the potential comodities are cassava, arenga, corn, coffee, peanuts, mung bean, pottato and various of vegetables. Crops harvest usually sale in raw or fresh material. It gives small income, and thats why focusing in this program of research are economic empowerment activity by using local knowledge. One of potential commodity that has a good prospect to develop is Aren. Aren itself has been used for producing sugar block and brown sugar.

Based on the results of the search-depth discussions with the craftsmen and a few people whom work in the government, it is important to do systematic efforts to improve palm sugar products to become one of the leading commodity products locally. Through technology transfer with training, the activities of the introduction of new products based on palm potential becomes possible to do refers to the development of superior products based diversification from this comodity. Usage the appropriate technology in the processing of local potential is considered able to contribute significantly in increasing the added value. In the processing of palm sugar they encounter various obstacles, especially in terms of firewood supplycontinuity. During this time they utilize the remnants of unused pieces of wood, and they are also regular traders to buy firewood at the price of IDR. 500.000, - / truck ankle (3 cubic). The average is used for a week to produce 15 kg of palm sugar. Negligence in the processing becomes important to avoid acidic sap that influence the outcome of palm sugar itself. For them the weather was very affecting the quality of palm sugar. The dry season or rainy season is not a major problem, except when the weather is erratic, sometimes hot sometimes rain. Occasional rain may lead to levels of sugar in the sap was considered good enough. Until now, the sustainability of palm sugar processing business which they run can make a positive contribution to the economic income

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families, because until now the demand is pretty good. Even that, the usage of appropriate technology to support their efforts as crucial, hygiene-related processing will certainly affect the final result, continuity processing and packaging that attract so marketing is not limited to traditional markets but could also encompass other market segments, especially in increasing the potential for agro-tourism in RejangLebong region. So the palm sugar or sugar ants can become a commodity in the regions as a typical product of this area, although in other places there is also a craftsman or palm sugar processing this. At least from the attractive packaging, hygiene processing can be sold as one packet of commercialization, and certainly will help increase the income of family economy.

The skills for activities of making sugar is the result of hereditary in its development in line with the development of interaction between residents and the transfer of knowledge that is more characterized by "scientific" primarily from government that have attention to the development of the business of making sugar palm in RejangLebong. As was done by the Center for Appropriate Technology Development LIPI Subang introducing palm sugar product diversification so that no other variants introduced by utilizing appropriate technology. Hopes the resulting product can appear "more fresh", and more attractive both in terms of presentation and of the packaging so that it can increase the added value of these products at a time can increase the income of society itself.

5. Potential Development of Aren based on Local Knowledge

Some diversification of the products we introduced are:

As already described at the previous sub section, palm processing into sugar are mostly done by migrants mainly from Java. The knowledge they applied and developed by considering natural resources in the region RejangLebong. Aren become one of the abundant natural resources.

1) Making printed palm sugar 2) Making ginger bread brown sugar 3) Preparation of liquid palm sugar ginger flavor

The skills they applied in Rejanglebong for the Aren processing get handed down from the elders or their parents when they were still in the place of origin (Java). Over time the interest of the population to cultivate palm sugar processing business has developed, children, siblings and even the locals RejangLebong. Even today, the craftsmen have managed to cultivate palm sugar crops in their garden to facilitate the coverage and quantity of production. The increase number of sugar producers are also followed by the development of knowledge processing and cultivation of palm plants in accordance with the realities of everyday life faced. This is similar to what is disclosed by Winarto (1998 and 2004) that the dynamic nature of knowledge itself, ever evolving and is accompanied proceed with the interaction between individuals, then with the learning process of each has been pushed knowledge that continues to be produced and reproduced, in which the process dunamika knowledge itself is a dialogue between two perceptions and different forms of learning between local scientific and causes and bring changes, modifications and variations are specified. In line with Winarto, Joshi, et al (2008: 84) emphasized that the knowledge gained from the "understanding and interpretation of the results of observations, experiences, as well as formal and informal education someone where knowledge itself is always in a state of growing along with the" development of observation, experience or the introduction of new innovations "that exist.

The public was enthusiastic about this training activity. This training is part of the technology transfer activities to the public, in addition to introducing some of the equipment that directly support the processing activities at the same time also can encourage the spirit of building a business based food processing local potentials. The activities start from the reality that societies and cultures everywhere are always in a state of change, whether changes occur quickly or slowly. Changes occur as natural as the physical state of knowledge and the society develops. Changes quickly become unnatural when society and knowledge can not understand the changes in social phenomena. Rapid changes generally occur because there are other communities that have different lifestyles to live together in a community or also because there is intervention from the outside through development programs implemented in public life (BambangRudita, 2003). The activities of Aren development in RejangLebong synchronize with the spirit of building a society where creative economic development in rural communities can not be separated from the principal elements such as technological development itself (Soelaiman, 1994). Technology becomes a tool used to promote changes in society, but the sophisticated technology as the device is always dealing with the local community with its values. The technology was not value free and often at odds with the values prevailing in the society. When technology is applied directly in the community regardless of social, cultural, religious, local knowledge held and believed the target communities so the difference causes the applied

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technology is not utilized properly even abandoned from the overall development program. Technology can not push changes or growth if not supported by local communities and institutions (social and cultural). Therefore, the development of local potential in this regard by introducing appropriate technology but still based on the social situation of cultural and local knowledge of the community itself. Local knowledge takes position as basic in developing new products that varied so that the technology was introduced into harmony with what they already have. The diffusion of mindsets introduced by the implementers with local knowledge of the community to produce a form of community (artisans palm sugar) that is open to knowledge from outside, then translated by the dose of the analytical community itself so that it can give to certain variants. In this case the introduction of technology takes a position as a trigger for creative endeavors undertaken the development of local communities. In this context also the public knowledge start from the results of dialogue between scientific knowledge from the outside, especially "FEA" and the knowledge society itself. The new knowledge gets purport locally by the community itself, which could be by using frame based on previous knowledge. The adoption and diffusion indicates a process of transformation of values and society's perspective of looking at the reality. Likewise with Aren development activities in RejangLebong, the existing local knowledge into a base which is then reunited with technology and new knowledge so that it can generate added value to the productaren itself ,where previously passing through the process of adoption, diffusion and dialogue of new knowledge and prior knowledge as its base. 6. The closing The activities of the introduction of appropriate technology to the public especially palm sugar producers be atrigger to increase value added one of the natural resources that exist. Local knowledge becomes the basis to develop potential local resources (palm) where knowledge and technology are introduced in accordance with the dynamics of the knowledge society itself. Introduction of technology in the Palm Sugar Industry not to shift any prior knowledge, but more toward strengthening knowledge through an interactive dialogical process in which the expectation is an increase in income generating public throughincreasing the added value of existing products. The cooperation between the Government of RejangLebong through BAPPEDA RejangLebong, P2TTG LIPI Subang and various business elements in RejangLebong particularly palm sugar producers are expected to contribute positively to the socio-

economic life of society. Not only is there an increase in people's income and added value of potential of existing areas, further also can provide strength in the community related to food security. Existing potential can be optimized through the use of appropriate technology. Aren become one of the entrances in an effort introduction of appropriate technology to the public. In this case the Aren is a potential coomoditythat has provided livelihood to many people, especially those who are engaged directly in the Aren-based businesses. Hopefully from what we do even depart from this little thing can bring change and a tremendous benefit for the community in particular and for the district RejangLebong general. Hopefully, the potential of its natural resources capable of providing Curup name or RejangLebong more widely known. Through the introduction of appropriate technologies for this palm-based food processing hopefully ideals in building local optimization potential can be realized. References [1] Agrawal, Arun. 1998. “Indigenous and Scientific Knowledge: Some Critical Comments”. Antropologi Indonesia: Majalah Antropologi Sosial Budaya Indonesia No 55 .Th XII. Januari-April 1998. Jurusan Antropologi, Fakultas Ilmu Sosial dan Ilmu Politik Universitas Indonesia. Hal 14-25. [2] Antweiler, Christoph. 1998. “Local Knowledge and Local Knowing, An Anthropological Analysis of Contested “Cultural Products” in the Context of Development”. Anthropos, Bd. 93, H. 4./6. (1998). Anthropos Institute. Hal. 469-494. [3] Badan Perencanaan Pembangunan Rejang Lebong. 2013. Kabupaten Lebong dalam Angka 2013. Rejang Badan Pusat Statistik Kabupaten Lebong.

Daerah Rejang Lebong; Rejang

[4] Burhanuddin, R. 2005. Prospek Pengembangan Usaha Koperasi dalam Produksi Gula Aren. Makalah; Jakarta. [5] Christenson, J.A and Robinson, JR, 1989, Community Development in Perspective, First Edition, Iowa State University Press, ISBN 081381474-X, Ames, Iowa. [6] Ditjen Perkebunan, 2004, Perkembangan Aren di Indonesia. Prosiding Seminar Nasional Aren. Tondano. 9 Juni 2004. Balai Penelitian Tanaman Kelapa dan Palma Lain. hlm 138 – 144. [7] Effendi, D.S., 2009, Aren, Sumber Energi Alternatif. Warta Penelitian dan Pengembangan Pertanian Tahun 2009. 31 (2) : 1 – 3.

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[8] Joshi, Laxman et al. 2008. “Sistem Sisipan: Pengetahuan Lokal dalam Wanatani Karet. Belajar dari Bungo: Mengelola Sumberdaya Alam di Era Desentralisasi. Bogor: Center for International Forestry Research (CIFOR). Hal. 83-100. [9] Rangkuti, Parlaungan Adil. 2011. Komunikasi Pembangunan dan Mekanisasi Pertanian. Bogor: IPB Press. [10] Rumokoi, M.M.M., 2004, Aren, Kelapa dan Lontar Sebagai Alternatif Pemenuhan Kebutuhan Gula Nasional. Prosiding Seminar Nasional Aren. Tondano. Balai Penelitian Tanaman Kelapa dan Palma Lain. [11] Soelaiman, M. Munandar. 1998. Dinamika Masyarakat Transisi: Mencari Alternatif Teori Sosiologi dan Arah Perubahan. Yogyakarta: Pustaka Pelajar. [12] Syakir dan D.S. Effendi, 2010, Prospek Pengembangan Tanaman Aren (Arenga pinnata MERR) untuk Bioetanol, Peluang dan Tantangan. Makalah disajikan dalam Workshop Peluang, Tantangan dan Prospek Pengembangan Aren untuk Bioetanol Skala Industri dan UMKM, Hotel Salak, Bogor, 21 Januari 2010. hlm 17 [13] Widyawati, N. 2011. Sukses Investasi Masa Depan dengan Bertanam Pohon Aren. [14] Winarto, Yunita T. 1998. “Hama dan Musuh Alami, „Obat dan Racun‟ Dinamika Pengetahuan Petani dalam Pengendalian Hama”. Antropologi Indonesia: Majalah Antropologi Sosial Budaya Indonesia No 55 .Th XII. Januari-April 1998. Jurusan Antropologi, Fakultas Ilmu Sosial dan Ilmu Politik Universitas Indonesia. Hal 53-68. [15] Winarto, Yunita T. 2004. Seeds of Knowledge: The Beginning of Integrated Pest Management in Java. New Haven, Connecticut: Yale University Southeast Asia Studies.

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Development of the Main Agroindustry Potential in Rejang Lebong District, Bengkulu Eki Karsani Apriliyadi a, Diki Nanang Surahman a, Hendarwin M Astro a* a

Center of Appropriate Technology Development, Indonesian Institute Of Sciences (IIS), Jl. K.S. Tubun No. 5 Subang 41213, Indonesia Tel.: +62-260-411478; fax: +62-260-411239. e-mail address :[email protected]

Abstract:The potential of Rejang Lebong region actually be a wealth of natural resources are exceptional, when processed and developed based on the implementation of appropriate technology will be superior to direct the potential areas that will provide increased welfare of society itself. Through the concept of community empowerment, the pattern of development of the potential of agro-industries / agribusiness is based on the ability of the region to provide the carrying capacity of its potential and wealth. Ability itself seen from two sides of the human resources (HR) and natural resources (SDA). Optimization of natural resources coupled with the optimization of human resources. This activity is aimed at generating income, improved quality of competitive self in the middle of the speed of globalization. Keywords: Agroindustry; development; RejangLebong; appropriate technology; 1. Introduction Rejang Lebong is one of the districts in Bengkulu Province, which has the potential wealth of natural considerable. Its potential is huge in supporting the development of agro-industry / agribusiness. The condition was supported by the availability of a wide area of 151.576 hectares are spread in 15 districts. Rejang Lebong is geographically located between two hills, west flanked by the Bukit Barisan Mountains and east by Mount Kaba with boundaries as follows: 1) The north is bordered by Lebong; 2) The south Regency Kepahiang; 3) Next to the western border with North Bengkulu and; 4) East of Musi Rawas regency of North Sumatra Province. Rejang Lebong region itself is situated at an altitude of less than 100-1000 meters above sea level to an altitude of 0-100 meters above sea level, covering an area of 2,621 H, 100-500 m above sea level covering 44 268 ha, altitude of 500-1000 m covering an area of 36 798 ha and at an altitude of 1,000 m up area of 67 899 ha. With a height as above, Rejang Lebong region into a fertile area and the potential for development of different varieties of food crops. Agriculture became the main activity of which relied upon by most of the population lives. Some varieties of crops that have been developed by population are: cassava, maize, sweet potatoes, peanuts, soybeans and green beans. In addition to the above crop is seeded vegetable crops are widely grown by farmers. Some of the many vegetable crops grown by farmers are: potatoes, red potatoes, cabbage/ Chinese cabbage, squash, kale and spinach. Not only that, the topography is hilly and fertile soil conditions the

development of plantation crops became very good potential for small scale and large scale. Some plantations of the population that has been developed, namely: palm, robusta coffee, rubber, coconut and cinnamon. The resulting coffee production has successfully penetrated the market to other areas in Sumatra, especially to Lampung. Although this region has been able to sell their coffee to the outside area, but the name raised as a producer of coffee actually not Rejang Lebong itself, but instead Lampung. For some people this is very unfortunate, why not Rejang Lebong which got the name as a producer of coffee. Especially for today's artisans palm palm sugar palm cultivation has been able to do, so that they are able to do their own breeding in orchards that they have. Previously they only rely on palm tree that grows in mountainous areas or hills around the area Rejang Lebong. The potential of Rejang Lebong region actually be a wealth of natural resources are exceptional, when processed and developed based on the implementation of appropriate technology will be superior to direct the potential areas that will provide increased welfare of society itself. Therefore, through the concept of community empowerment, the pattern of development of the potential of agroindustries / agribusiness is based on the ability of the region to provide the carrying capacity of its potential and wealth. Ability itself seen from two sides of the human resources (HR) and natural resources (SDA). Optimization of natural resources coupled with the optimization of human resources. This activity is aimed at generating income, improved quality of competitive self in the middle of the speed of globalization. Economic potential is given the opportunity to be optimized with the strength of the community. This hope must be realized in a practical level, it is not only limited to discourse that live in the level of ideas. One

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manifestation of this realization is poured through a participatory approach as the concept of empowerment is rotated with the intended value is independence. Local communities are stimulated, encouraged, and joined into applicative activities through training as one form of technology transfer. The concept of sustainable development is emphasized for more coloring expectations to be addressed. The development potential of the region is not limited to optimization, but more than that, there is a genuine effort to ensure the sustainability of what was invested at the start to bring hope to the ongoing sustainability assurance can be extracted and utilized optimally. Therefore started from the expectations and where we are going, people should be placed as the subject or the main actors in the construction activity itself and not become an object. Community directly stimulated, encouraged and joined on the existing problems to better understand the potential of its own territory. Various skills are granted in accordance with the potential of its territory be used as a trigger for further creation of further understanding to arrive at the stage of development (agro-industry) region based on selfreliance and ability alone. Skills that can be transferred through a variety of training as one form of technology transfer. Started from the above presentation development missions executed based on the economic empowerment of the people through the development of a democratic economic system. Small and medium entrepreneurs and cooperative the main target of the target based on market mechanism, based on a resource that is productive, independent and forward. Another mission is to strengthen the economic foundation of the community-based agro-industry and develop it so that it becomes a commodity and seeded in the future. Agricultural potential, farming, and other potential today remains largely marketed in the form of further unprocessed primary. Therefore, economic empowerment activities in Rejang Lebong focused on the development of agro-industries to target the development of small and medium enterprises based on local potential. One way to develop agro-industry in this district is to support the implementation of the results of research and development stemming from R & D institutions in Indonesia. Appropriate technology can be applied in an increase in value added products and the productivity of society and is part of the empowerment and development of small and medium enterprises in Rejang Lebong. 2. Issues

subsistence lifestyle, also affected by the condition of the area and human resources (HR). Low levels of mastery of technology led to an abundance of local potential is not utilized optimally. These conditions are as follows: 1) Low levels of knowledge lead to the low level of use of technology that can support as a tool in their daily lives; 2) Low levels of education result in a lower quality of available labor; 3) The geographical conditions hilly result in an uneven distribution of the population. 4) The shortcomings of the government's demands great attention both from the central and regional levels to the development of Rejang Lebong, particularly in rural areas still lag behind in development. On the implementation of development policy, there is still a attraction between the development orientation that prioritizes "growth" with the construction of more development priority "equalization". This gave rise to the impression of discourse "people's economy" less followed by policies and programs that are less precise adjustment. It is feared will lead to the development of agribusiness is only enjoyed by a handful of people in a small amount, do not touch the community at large. 3. Analytical Framework Society and culture anywhere is always in a state of change, whether changes occur quickly or slowly. Changes occur as normal because of the knowledge and the physical state of that society develops. Changes quickly become unnatural when society and knowledge can not understand the changing social phenomena. Rapid changes generally occur because there are other communities that have different living patterns of living together in a community or because there is intervention from the outside through development programs implemented in public life (Bambang Rudita, 2003).

Development in rural communities can not be separated from the principal elements such as technological development itself (Soelaiman, 1994). Technology becomes a tool used to promote changes in society, but the sophisticated technology as the device is always dealing with the local community with its values. The technology was not value free and often at odds with the values prevailing in society. If the technology is applied directly in the community without taking into account the social, cultural, religious and other community objectives then it causes a difference in the technology applied is not put to good use instead left out even against the overall development program.

Development problems faced by the people of Rejang Lebong in addition to social, cultural and economic conditions of society are realized in a 1st International Conference on Appropriate Technology Development 2015, Bandung, October 5th-7th, 2015 | 140

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Technology can not encourage change or growth if not supported the participation of local communities and institutions (social and cultural). In order for the technology transfer process goes as expected according to Khalil (2000) need to pay attention to the three concepts of technology transfer. The third concept is the local manpower for technology transfer, dissemination or diffusion and adaptation and technology development (Gumbira, 2000). On the other side should also be noted the readiness of society to accept the technology and certainly prepared matter of expediency technology itself to the community. Dalam efforts to accelerate economic growth and development capabilities in the development of sustainable communities then develop the economic potential through the empowerment and development of Small and Medium Enterprises (UKM) is regarded as one of the appropriate strategy based on the potential for a locally owned. At the time of economic crisis hit the country, UKM has proven durability that is expected to become the foundation of the national economy. Thus the development, strengthening and expanding the role of UKM in the economy of the community occupies a strategic position in the face of conditions that are susceptible to change.Development of Small and Medium Enterprises (UKM) based on local potential is expected to be a strategy that can help the optimization potential of agro-industries in the target areas, especially in areas that still rely on natural resources to drive economic growth. 4. Hypothesis 1) If the human resource potential enhanced to optimize natural resources are owned through training and coaching, the standards of living can be improved. 2) The rate of economic growth can be achieved by increasing public participation in economic activities. 3) Productivity can be improved if the public community technology capabilities enhanced. 4) Successful implementation of technology and technology transfer can be achieved when a community technology enhanced management capabilities. 5. Methodology The program implemented by utilizing executioncommunity based activity, society is positioned as development actors and not as an object. Basically its implementation aimed at creating self-reliance of local communities to organize their own social and economic. Its implementation is also supported by action research method, a method that more directly carry out activities can be enjoyed by the community. With this method is expected to particular problems that exist in society can be solved, where people directly

involved to recognize, understand the problems and can find solutions. Studies to support community economic development activities can be carried out with the participation of the observation method, between researchers and community members as users and recipients of technology. Researchers studied the program and the community directly from within the community. In practice also involves the related institutions including the Bappeda Rejang Lebong supported by the district government Rejang Lebong.

6. Implementation and Results of Activities 6.1 Implementation of Appropriate Technology of food processing technology implemented in the village of Air Meles Atas Rejang Lebong. In general, Air Meles Atas is palm sugar farmers and craftsmen. Agricultural products are generally sold directly obtained in raw form. While the results of tapping sap from palm trees they make palm sugar they sell directly to collectors. Implementation

Palm becomes one of the potential natural resources are very abundant, and most have been harnessed into palm sugar. When this has been a lot of people who can cultivate the palm trees in the gardens they have. Previously they only rely on palm trees that grow naturally in the forest. Farming activities carried out in an effort to develop the business of making palm sugar where current demand is increasing. Marketing is up to the various regions in Sumatra, especially from West Sumatra where palm sugar is a raw material for making soy sauce. Based on the results of the search-depth discussions with the craftsmen and a few people sitting in the seats of government, it is very important to do systematic efforts to improve palm sugar products to become one of the leading commodity products locally. Through technology transfer with training, the activities of the potential introduction of new products based on palm becomes possible to do refers to efforts to diversify the development of superior products based on the potential of this palm. Touch the appropriate technology in the processing of local potential is considered able to contribute significantly in increasing the added value. Most of the craftsmen are the settlers from Java and direct descendants, so that when we were there, as if we were in the rural areas of Java. They brought palm sugar processing technology into the area of Bengkulu. The palm trees as a source of sap they had cultivated his own. So most of the craftsmen have had its tree sap on their own, no longer have to go to the forest to collect the sap from palm trees. On average the master craftsmen 100-150 palm trees. In an average day they are able to produce palm sugar as much as 5 kg-15 kg depending on the amount of raw material processed.

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The products they produce then they are selling to the container / toke at a price of Rp. 10.000, - Rp. 11.000, -. At the average market price ranges from Rp. 15,000-RP. 17.000, -. In the result they commonly produce sugar gambier, namely palm sugar that is molded into small pieces while they get used to print it in a coconut shell mold. Besides regular sugar they utilize its sap for sale, it could be a fresh drink. In fact, according to them sometimes there that makes it a wine. This is a concern for many people, because if it is not anticipated that it could interfere with the moral values of local communities. According to some people usually buy the sap is processed into wine usually is people from outside the area usually Batak people. In spite of it all, palm sugar processing business into a promising business area even for those businesses is a major effort in addition to farming. Here an artisans of palm sugar has been managed in groups of craftsmen who later joined the Gapoktan. For those with flocking way many benefits they can get, such as: 1) Easy to get help from the government, either aid or assistance cash capital business equipment; 2) Easy to get guidance related to knowledgerelated knowledge processing palm sugar. The group was formed on the initiative of the government represented by the FEA. In addition they also expect that with this group they always get guidance and assistance until they are able to increase productivity so as to increase prosperity. They expect that they are able to produce products other than sugar alone printing. In fact they have had knowledge of the manufacture of sugar ants, but for those processing are considered more complicated than ordinary sugar palm print. In addition the proceeds are considered not very profitable. In the processing of palm sugar they encounter various obstacles, especially in terms of continuity of supply of firewood. During this time they utilize the remnants of unused pieces of wood, and they were also used to buy firewood to merchants at a price of Rp. 500.000, - / truck ankle (3 cubic). The average use for a week to produce 15 kg of palm sugar. Negligence in the processing becomes important to be taken to avoid acidic sap that influence the outcome of the palm sugar itself. For them the weather was greatly affect the quality of palm sugar. According to them the dry season or wet season is not a major problem, except when the weather is erratic, sometimes hot sometimes rain. Occasional rain may lead to levels of sugar in the sap was considered good enough. Until now, the sustainability of palm sugar processing business which they run can make a positive contribution to the economic income families, because until now the demand is pretty

good. Even so a touch of appropriate technologies in support of their efforts as crucial, related hygiene treatment will certainly affect the final result, the continuity of the processing and packaging of interest so that marketing is not limited to traditional markets but also can capture a market segment others, especially in increasing the potential for agrotourism in Rejang Lebong region. Palm sugar and sugar so that the ants can become a commodity in the regions as a typical product of this area, although in other places there is also a craftsman or palm sugar processing this. At least from the attractive packaging, hygiene processing then removed and sold as a package commercialization, we think it can help increase the income of their family economy. In order to develop the potential of this agro several technologies are introduced. Potential equipment appropriate technology that is implemented in food processing, among others, is a tool plastic sealer as a tool for the packaging of processed food that is ready-made and tool cup sealer as a tool for closing plastic beverage containers are finished and digital scales, a tool chopper crackers used to slice potatoes and potato chips are used as display devices of palm sugar. All the equipment used for the purposes of food processing training to the cadres of participants in the target area. 6.2 FoodProcessing Training In accordance with the potential of agro in the target areas of training conducted as a follow up of the transfer of technology to the public. Based on the potential of the region and the public demand as well as coordination with the Agency, some food processing training package which was introduced consisting of : 1) 2) 3) 4) 5) 6)

Making printed palm sugar Making gingerbread sugar ants Preparation of liquid palm sugar ginger flavor Making a variety of crackers (crackers onion, carrot crackers, and potato chips red) Making various flavors of potato chips red Making cocktails

Based on the results of assessments and discussions with the local community and in the training BAPPEDA enter food processing activities outside the base of palm. It departs from the local conditions very remarkable that still needs a touch of technology in order to provide added value. The public was enthusiastic about this training activity. This training is part of the technology transfer activities to the public, in addition to introducing some of the equipment that directly support the processing activities at the same time also can encourage the spirit of building a food processing business based on local potential that exists.

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The training was attended by 15 participants both male and female. The participants came from Air Meles Atas villagers. Once the training is finished then some ingredients for food processing BAPPEDA handed over to the Rejang Lebong. This meant that the BAPPEDA continue to provide guidance and monitoring of the results of training activities in Air Meles Atas community. 6.3 Development andAssistance Group As the next steps, besides training, development will be conducted and support groups that have been included in the training of food processing. In an effort formation and development of enterprises, business groups lent some equipment that is associated with the business to be run. All equipment of appropriate technology that is implemented then stored in the business group formed. The hope that they are able to utilize the equipment for the production of households and enterprises can continue to grow. This meant that the BAPPEDA continue to provide guidance and monitoring of the results of training activities in Air Meles Atas community. Mentoring and monitoring is very important to do given the technology transfer process is not completed only until the training is over. Keeping the spirit and rhythm of business activities becomes important as fuel to promote and develop the business to be a positive impact for the culprit in particular and for society in general Rejang Lebong. In addition to Bappeda and local SKPD, our team P2TTG LIPI Subang strengthening against the trainees. This meant that the technology transfer activities do not stop, worried when no reinforcement of what has been trained to be forgotten. Our hope hopefully the knowledge and technology transfer to the public can be useful and beneficial for the economy. Besides the potential of local resources can be optimized and able to lift up the name of the District Curup as expected by the local government. 7. Conclusion From the results of activities carried out in the field, it can be concluded that: 1) The food processing activities, has formed several cadres otherwise be able to popularize the results of appropriate technology training. 2) From the results of the training activities of food processing has been seen that each of the participants have an interest in different business sectors, in addition also means that there are diversified fields of business carried on. 3) Touch technology is indispensable in efforts to increase the added value of natural resources based locally.

4) The post-harvest handling equipment to diversify processed products introduced are still desperately needed by the people of Air Meles Atas Rejang Lebong and society in general. 5) Equipment appropriate technology of food processing can lead to the formation of other small businesses in post-harvest processing of commodity-based local that has not been used optimally. 6) In an effort to equitable development and economic growth, the need for sustainable agro-industry development directed to rural areas. 7) It is necessary to support the long-term policy and sustainable agro-industrial development of the government, especially local government to stimulate and develop small and medium enterprises based on the development potential of agro-industries in the target area. 8) It is necessary to support local government policy and the equal opportunity to obtain the results of development especially in meeting the primary needs of the community. 9) Need for attention, support continued to organize and maintain equipment appropriate technology in order to maintain the continuity of the business. Bibliography [1] Adimihardja, Kusnaka. 1999. “Mendayagunakan Kearifan Tradisi dalam Pertanian yang Berwawasan Lingkungan dan Berkelanjutan”, dalam Petani: Merajut Tradisi Era Globalisasi. Bandung: Humaniora Utama Press. [2] Badan Perencanaan Pembangunan Rejang Lebong. 2013. Kabupaten Lebong dalam Angka 2013. Rejang Badan Pusat Statistik Kabupaten Lebong.

Daerah Rejang Lebong; Rejang

[3] Gumbira E., dkk. 2001. Manajemen Teknologi Agribisnis. Jakarta: Ghalia Indonesia. [4] Soelaiman, M. Munandar. 1998. Dinamika Masyarakat Transisi: Mencari Alternatif Teori Sosiologi dan Arah Perubahan. Yogyakarta: Pustaka Pelajar. [5] Suyuti, Nasrudin. 2005. Masyarakat Tani dan Pemanfaatan Inovasi Teknologi (Suatu Tinjauan Sosial Budaya) yang dipresentasikan dalam Seminar Nasional dan Ekspose Hasil Penelitian, 18-19 Juli 2005. Kendari. Balai Pengkajian Teknologi Pertanian

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Yield Risk Assessment in Nutrient Film Technique for Pakcoy (Brassica rapa L.) Hydroponic Growing System Using FMEA and AHP Approach: A Case Study Yusuf Andrianaa, Eko Kuncoro Pramonoa, Cahya Edi Wahyu Anggaraa, Aidil Haryantoa, Ignatius Fajar Apriyantoa a

Centre for Appropriate Technology Developmnet, Indonesian Institute of Sciences Jl. KS Tubun No.5 Subang, West Java 11420, Indonesia Tel.: +62-260-412878 E-mail address: [email protected]; [email protected]

Abstract: Risk is an intrinsic part of agricultural management system. Crop yield in agricultural production is uncertain both quality and quantity. Several variables affect crops yield, such as: light, nutrient, temperature, humidity, pest and disease, etc. It also occurs in NFT hydroponic growing system. To mitigate yield failure or decreasing quality and quantity in NFT hydroponic growing system, it is needed risk assessment to formulate strategy to mitigate the potential yield risks failure. The purposes of this study were (a) identify potential yield risk in NFT for Pakcoy (Brassica rapa) hydroponic growing system, and (b) formulate strategy to mitigate production failure or decreasing crop yield based on prioritized the potential yield risk. The methods of this study were failure modes and effect analysis (FMEA) and analytical hierarchy process (AHP). The FMEA was implemented to point out the probable failure modes. Then the AHP methodology was used to prioritize the most potential yield risk factor to formulate strategy in NFT technique yield risk mitigation. The results showed that there were several potential risk factor sources that affected crop yield in NFT hydroponic growing system such as seedling, nutrient, pest and disease, environment, aeration, and water. The most important potential yield risk factor in NFT system was water. Keep the water circulated, water level, flow rate and nutrient concentration is key factor to successful growing Pakcoy (Brassica rapa L.) hydroponically using NFT system. Keywords: risk assessments, nutrient film technique, FMEA, AHP 1. Introduction Like a traditional farming, hydroponic farming is a financially risky occupation. On daily basis, farmers are confronted with an ever-changing landscape of possible price, yield, and other outcomes that affect their financial returns and overall welfare. The consequences of decisions or events are often not known with certainty until long after those decisions or event occurs, so outcomes may be better or worse than expected (Harwood, et al. 1999). Understanding risk is a key element in helping farmers makes better decisions in risky situations. Risk is uncertainty that affects an individual‟s welfare, and is often associated with adversity and loss (Bodie and Merton, 1998). Hardaker et al. (1997) divided risk source into five general categories: production or yield risk, price or market risk, institutional risk, human or personal risk, and financial risk. Yield risk is one of risk sources component in farming and is an important issue in agriculture management system. Managing risk in agriculture has been reported in several studies; Harwood et al. (1999) did a research on the risks confronted by crop farmers. According to their result, there are three kinds of risks which are institutional risk, yield risk and price risk related to agriculture. In general producer of major crops tend to be more concerned about production risk; Archer et al. (2003) proposed a stochastic budgeting example using field plot data to illustrate common

sources of risk and methods producers use to deal with risk; Xiang et al. (2012) evaluated crop distribution in Yunnan, China as implication for crop engineering risk and proposed a standard measure of crop engineering risk that is based on the expected loss of crop yield; Elagib (2014) developed and applied a drought risk index for food cop yield in Eastern Sahel. But risk management in specified field such as hydroponic is very limited reported. Risk analysis is a part of risk management. That is, the four main aspect in risk management involve (1) identifying potentially risky event, (2) anticipating the likelihood of possible outcomes and their consequences, (3) taking action to obtain a preferred combination of risk and expected return, and (4) restoring (if necessary) the firm‟s capacity to implement future risk-planning strategies when distress condition have passed (Hardaker et al., 1997). Marimin and Maghfiroh (2010) limited risk analysis study at least consist of risk identification, risk and decision investigation, and risk management action implementation to overcome risk failure. There are several techniques developed to perform risk analysis to mitigate the failure. The technique Failure Modes and Effect Analysis (FMEA) is one of the most widely used risk analysis tool. Bowles and Pelaez (1995) mentioned that FMEA was originally developed for systematic analysis of failure modes and its subsequent effect for the defense related product particularly in the aviation

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sector. This FMEA was first proposed by National Aeronautics and Space Administration (NASA, U.S.A) in 1960. Then, it was adopted and promoted by Ford Motor in 1977 (Maheswaran and Loganathan, 2013). Today, FMEA has been adopted in wide spectrum of fields such as chemical, aerospace, nuclear, automobile, mechanical, semiconductor, (Mandal and Maiti, 2014; Maheswaran and Loganathan, 2013), and service industries like software industry (Shawulu, 2012), But very limited study which discussed FMEA application in agriculture. The major objective of application of FMEA is the identification of potential failure modes system component, evaluating their causes and their subsequent effect on the system behavior, and as a result determination of the ways to eliminate or reduce either the chances of occurrence or severity or increase the detectability of the particular failure mode (Mandal and Maiti, 2014). Traditionally, FMEA the risk computation of different failure modes using FMEA has been done by developing risk priority number (RPN). RPN is the value obtained by the product of three components, i.e. the occurrence probability of failure mode (P), the severity of the failure mode (S), and the detectability of failure mode (D). Higher the value of the RPN higher is the risk associated with corresponding failure mode. In this method expert will give their preference level based on the scale importance to each failure modes. This RPN method has been criticized due to its limitation. So that, many authors proposed FMEA with multi criteria decision making (MCDM) techniques. There are many studies that combining FMEA and MCDM i.e. analytical hierarchy process (AHP) for assessing risk. Shahrabi and Shojact (2014) applied FMEA and AHP in lean maintenance; Hu et al. (2009) evaluated risk of green components to hazardous substance using FMEA and fuzzy AHP; Kamble and Quazi (2014) used FMEA to identify the potential failures of shell molding process and prioritized risk factor using AHP. In this study we used FMEA and AHP to assess yield risk in very famous hydroponic technique, nutrient film technique, for pakcoy (Brassica rapa) hydroponic growing system. The purposes of this study were (a) identify potential yield risk in NFT for Pakcoy (Brassica rapa) hydroponic growing system, and (b) formulate strategy to mitigate production failure or decreasing crop yield based on prioritized the potential yield risk. 2. Methods In this study we used FMEA for identification of yield risk factor in NFT for Pakcoy (Brassica rapa L.) hydroponic growing system. AHP methodology is used to prioritizing yield risk factor. The steps of this study are showed in Figure 1.

2.1 Failure modes and effect analysis (FMEA) FMEA is used to identify potential yield risk in this study. FMEA employs Risk Priority Number (RPN) to measure the risk and severity of failures (Rhee & Ishii, 2003). RPN is an index that can represent the degree of risks that a product may possess. It consists of three indicators: Occurrence (O), Severity (S), and Detection (D). Basically FMEA consist two stages; the first phase is to identify the potential failure modes and decide the value of Severity, Occurrence, and Detection. In the second phase, the manager should make recommendations for correct actions, and the RPN needs to be recalculated after correct actions (Su and Chou, 2008). Table 1 described scale rating for Occurrence, Severity, and Detection. The detailed description of the FMEA creation process can be found in the work of McDermott, Mikulak, and Beaugerard (1996). Begin Begin

Failure Failure record record

What-If What-If Analysis Analysis

Identification Identification of of probable probable failure failure mode mode

FMEA

Identification Identification of of risk risk factor factor Prepare Prepare and and evaluate evaluate the the questionnaire questionnaire for for AHP AHP Determine Determine the the weight weight for for each each risk risk factor factor

Consistency Consistency check check for for weights weights

Prioritized Prioritized risk risk factor factor

AHP Strategy Strategy formulation formulation for for mitigate mitigate the the risk risk

End End

Figure 1. Step of this research

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2.2 Analytical hierarchy process (AHP) In This study we used AHP (Saaty, 1987) to prioritise of potential yield risk factor based on risk factor identification using FMEA method. Both qualitative and quantitative criteria can be combined with this method to determine useful weight-age information which provides a mechanism of decision making.

and criteria j, such that ai,j = 1, ai,j =1/ai,j and ai,j 0. Then the weight vector

𝑤𝑖 = 1/𝑛 𝐴𝑗 = 1/𝑛

𝑛 𝑗 =1 𝑎′𝑖,𝑗 with

𝑎′𝑖,𝑗 = 𝑎𝑖,𝑗 /𝐴𝑗

𝑛 𝑖=1 𝑎1,𝑗

and (1)

To check the consistency of the pair-wise comparison and credibility of weights the consistency ratio (CR) is calculated as:

In the first step of AHP, a model is structured in hierarchy using objective, criteria, and decision alternatives. After defining the hierarchy, all alternatives and criteria are compared one to one in order to determine the relative importance of criteria within each level. The pair-wise comparison is performed according to their level of influence and based on the specified criteria in the higher level. It starts from the second level and finishes to the alternative levels at the bottom. A standardized comparison scale is used to find the relative importance of the criteria exhibit in the Table 2.

𝐶𝑅 = 𝐶𝐼/𝑅𝐼 with 𝐶𝐼 = (𝜆𝑚𝑎𝑥 − 𝑛)/(𝑛 − 1)

(2)

Where 𝐶𝐼 is the consistency index for the matrix; 𝑅𝐼 is the random index for different n is available in Saaty (1990). Table 3 shows the value of random index (𝑅𝐼) for matrices of order 1-10 obtained using sample size of 500 [47]. The smaller (<1) the value of CR the better is the judgments of decision maker indicating the pair-wise comparison matrix and the computed weights are reasonable. Larger values require the decision maker to reduce inconsistencies by revising judgment.

The result of the pair-wise comparison on n criteria can be summarized in an n x n evaluation matrix A in which every element ai,j(i,j = 1,2,3…..n) is the intensity of relative importance between criteria i

Table 1. Scale rating for Occurrence (O), Severity (S), and Detection (D) Occurrence Grade of Occurrence

Severity Grade of Severity

Number of times

Score

Extreme

More than one per day/ every 1 to 2 days

10,9

Extreme

Very High

One per every 2-4 days/per one week One per every 1-2 weeks/ 2-4 weeks

8,7

Very High

6,5, 4

High

One per every 1 to 3 month/ 3-6 months One occurrence per more than a year

3,2

Moderate

1

Low

High

Moderate Low

Effects of Severity Extreme failure to yield crop High failure to yield crop Moderately failure to yield crop Little failure to yield crop Very low effect failure to yield crop

Detection Grade of Detection

Likelihood of Detection

Scores

9

Extre me

Extreme chance for the system to detect the failure

9

7

Very High High

Very high chance for the system to detect the failure High chance for the system to detect the failure

7

Moder ate Low

Moderate chance for the system to detect the failure Low chance for the system ti detect the failure

3

Scores

5

3 1

5

1

Table 2. Scale of relative importance of each potential risk factor Relative Explanation Definition Equal importance Moderately important Strongly more important Very strongly more important Extremely more important Intermediate values Reciprocal of above

importance 1 3 5 7

Two activities contribute equally to the objective Experience an judgment slightly favor one activity over another Experience and judgments strongly favor one activity over another An activity is strongly favored and its dominance is demonstrated in practice 9 The evidence favoring one activity over another 2,4,6,8 When evidence favoring one activity over another is of the highest possible order of affirmation If activity i has one of the above non-zero numbers assigned to its when compared with activity j, then j has the reciprocal value when compared with i

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Table 3. Average random index (RI) at matrix size (Saaty, 1994) N

1

2

3

4

5

6

7

8

9

10

RI

0

0

0.52

0.89

1.11

1.25

1.35

1.4

1.45

1.49

1-10 obtained using sample size of 500. The smaller (<1) the value of CR the better is the judgments of decision makers indicating the pair-wise comparison matrix and the computed weights are reasonable. Larger values require the decision maker to reduce inconsistencies by revising judgment.

greenhouse in DFS is showed in Figure 2 and Figure 3. The NFT installation was put in the greenhouse. 20 cm

PVC pipe

3. Result and Discussion 3.1 Case Study: Hydroponic Growing System in Dawuan Field Station, Subang Regency, West Java Dawuan Field Station (DFS) is a transfer technology unit under Centre for Appropriate Technology Development (CATDev) coordination. There is a hydroponic farm model in DFS which consist of a greenhouse and several sets of equipment for vegetables hydroponic growing system. NFT hydroponic system is one of model that developed in DFS. The schematic diagram NFT installation and

400 cm

35 cm

Nutrient solution stock tank Aquarium pump

Figure 2. Schematic diagram of NFT system that installed in DFS.

Figure 3. Size and schematic diagram of greenhouse in DFS The NFT installation has made from polyvinyl chloride

(PCV) pipe which length was 4 m. There were six level in NFT installation with slope about 1 degree. The distance between the plant holes was 20 cm. The nutrition solution was circulated with 90 watt power of aquarium pump. While, the greenhouse was made using light steel, the dimension was 7.30 × 13.67 × 5.6 m. Insect net was installed around the greenhouse to protect it from pest.

nutrient solution passes. The trough containing the plant roots is set on slope (usually about 1 %) so that the nutrient solution is introduced at the top of the trough can flow from the top to the lower end by gravity at recommended flow rate of ¼ gallon (1 liter) per minute (Jones, 2005). Almost vegetables could be grown using NFT system. One of popular vegetables that are reported successful growing with NFT system is pakcoy (Brassica rapa L.).

Nutrient film technique (NFT) is a technique for growing plant hydroponically in which the plant roots are suspended in a slow-moving stream of nutrient solution. NFT was developed by Allan Cooper in 1970s. In NFT system, plants roots are suspended in a trough, channel, or gully (trough will be the word used from this point on) through which

Pakcoy (Brassica rapa L.) was originated from China. Growing area that is suitable for this plant is 100-500 m above sea level. Pakcoy is well growth in high-land or low-land area and can be cultivate through year (Sutirman, 2010). In this study, we observed growing condition in DFS around August 2015. Based on BMKG data, in August 2015 the

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weather in Subang regency was glorious-cloudy; the average temperature was 23-33degree Celsius; and the rainfall range was 0-20 mm per month. This condition should be suitable for pakcoy growing. 3.2 Identifying Potential Yield Risk Factor of FMEA In this study we classified yield risk source factor in pakcoy growing in six categories. There were seedling, nutrient, pest and disease, environment,

aeration, and water. There were also yield risk sub factors in this risk identification. Cause-effect diagram was used to represent potential source of yield risk that may cause decreasing or failure of pakcoy yield crop in NFT (Figure 4).

Figure 4. Caused-effect diagram of potential risk factor related possibility of decreasing/failure yield crop Seedling might cause decrease or failure of yield. The quality of seed of course affect the crop yield, also seedling technique, and germ age movement to the NFT system. The movement of germ must be in appropriate age, about 2 weeks, or the pakcoy have had four leaves. Nutrient also a part of yield risk source. Inappropriate concentration of macronutrient (N, P, and K) and micronutrient (Ca, S, Fe, Mo, etc.) could cause decreasing crop yield. Pest and disease factor also affect pakcoy yield crop. In a greenhouse DFS, there are hole with diameter about 5-10 cm. Insect can come in greenhouse and make infestation in the pakcoy plant. Environment factor such as, temperature, humidity, sun lighting, velocity of wind also affected yield crop, because each variety plant has requisite condition itself. The score of Occurrence (O), Severity (S), and Detection each potential yield risk is showed in Table 4. The highest score of potential yield risk source is flow rate of water with RPN score is 105. Water flow rate affected the yield because: flow rate water retaliated with nutrient distribution and dissolved oxygen which is needed root to absorb the nutrient and oxygen. Jones (2005) recommended flow rate is about ¼ gallon (1 liter) per minute.

3.3 Prioritizing Potential Yield Risk Factor of AHP Potential yield risk factors were prioritized using AHP methodology. There are three hierarchy levels in AHP diagram in this study which is showed in figure 5. First, decided the goal of AHP decomposition matrix that was assessing of yield risk factor in NFT for pakcoy hydroponic growing system. Second, made the criteria risk assessment; there were Occurrence (O), Detection (D), and Severity (S). Third, adopted the alternatives potential yield risk in NFT based on risk factor identification in FMEA method. Based on expert judgment, the most importance criteria to assessing yield risk was Detection (D) with score 0.731. Detection the risk is more important than taking action to ward off failure. So in the agricultural risk detection the risk is important in case to mitigate yield risk factor. Water is most yield risk factor source which might be affected to the crop yield. To mitigate the yield risk; keeping the water level, flow rate and concentration nutrient were key factor to successful growing Pakcoy (Brassica rapa L.) hydroponically using NFT system.

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Table 4. The score of Occurrence (O), Severity (S), and Detection each potential yield risk Score No 1

Occurrence

Severity

Detection

RPN

Inappropriate seedling technique

2

2

5

20

Low-grade seed quality

1

5

5

25

Inappropriate germ age that moved to tunnel

3

3

3

27

Inappropriate concentration of macro nutrient (N,P,K) Inappropriate concentration of micronutrient (Ca, S, Fe, Mg, B, Mn, Zn, Mo etc,)

2 2

5 5

3 3

30 30

Inappropriate nutrient supply

5

3

5

75

There are holes on green house that caused insect come in

3

3

3

27

There is no sterilization system in green house

3

3

3

27

Humidity is not fit with pakcoy requirement

3

3

1

9

Sun lighting is not fit with pakcoy requirement

3

3

1

9

Temperature is not fit with pakcoy requirement

3

5

1

15

Wind velocity that caused collapse

5

3

3

45

Inappropriate of dissolved oxygen (O2)

2

3

3

18

Inappropriate of slope tunnels

4

3

3

36

Inappropriate of CO2 requirement for respiration

2

2

3

12

Level of water in the tunnels

6

3

3

54

Debit of water

6

5

3

90

Flow rate of Water

7

5

3

105

Leaking in the tunnel

7

3

3

63

SEEDLING

2

NUTRIENT

3

PEST AND DESEASE

4.

5.

6

Yield Risk Factors in NFT

ENVIRONMENT

AERATION

WATER

Figure 5. Hierarchy level in AHP risk assessment

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4. Conclusion There were several potential yield risk sources in pakcoy (Barissca rapa) hydroponically growing system, such as were seedling, nutrient, pest and disease, environment, aeration, and water. The most important potential yield risk factor in NFT system was water. Keep the water circulated, water level, flow rate and nutrient concentration is key factor to successful growing Pakcoy (Brassica rapa L.) hydroponically using NFT system.

[8]

McDermott, R., Mikulak, R., Beauregard, M. 1996. The basics of FMEA. USA: Productivity

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Maheswaran, K., T. Loganathan. 2013. A Novel Approach for Prioritizing of Failure modes in FMEA using MCDM. International Journal of Engineering Research and Applications (IJERA) 3 (4), pp. 733-739

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Mandal S., J. Maiti. 2014. Risk Analysis Using FMEA: Fuzzy Similarity and Possibility Theory based Approach. International Journal of Expert System with Applications 41, pp. 3527-3537.

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Marimin, N. Maghfiroh. 2010. Application of Decision Making Technique in Supply Chain Management. IPB Press: Bogor, pp.138 (in Indonesian)

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Rhee, S.J., Ishii, K. 2003. Using cost based FMEA to enhance reliability and serviceability. Advanced Engineering Informatics, 17.pp. 179-188

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Saaty, T.L. (1987). The analytic hierarchy process-what it is and how it is used. Journal of Math Modeling 9( 3), pp. 161-176

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Saaty, T.L. (1994). Highlight and critical points in the theory and application of the Analytic Hierarchy Process. European Journal of Operational Research ,74. pp. 426-447

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Shawulu, H.N. 2012. Software Failure Analysis at Architecture Level using FMEA. International Journal of Software Engineering and its Applications 6 (1)

Acknowledgements The authors would like to thank Centre for Appropriate Technology Development (CATDev)Indonesian Institute of Sciences (LIPI) for financial support of this work by Thematic Program 2015. We are very grateful to Dr. Ir. Yoyon Ahmudiarto, M.Sc. IPM (Head of ICATDev-LIPI), Rima Kumalasari, MM, Dr. Rislima Sitompul, Mr. Sukirno, Mr. Agusto WM, Mr. Ismu Tribowo, Mrs. Nurhaidarahman and the other member of the Field Station Development Research Group for their input, guidance, and help. References [1]

Archer, D.W., J.L. Pikul, Jr., W.E. Riedell. 2003. Analyzing Risk and Risk Management in Cropping Systems. Dynamic Cropping System Symposium. pp. 155-164.

[2]

Bodie, Z., R. C. Merton. 1998. Finance. Upper Saddle River, NJ: Prentice Hall

[3]

Hardaker, J.B., R.B.M. Huirne, J.R. Anderson. 1997. Coping with Risk in Agriculture. CAB International, Wallingford, UK, pp. 274

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Harwood, J., R. Heifner, K. Coble, J. Perry, A. Somwaru. 1999. Managing Risk in Farming: Concept, Research, and Analysis. Agriculture Economic Report, No. 774, ERS USDA

Shahrabi, M., A. A. Shojaei. 2014. Application of FMEA and AHP in Lean Maintenance. International Journal of Modern Engineering Science 3(1), 61-73

[17]

Hu, A. H., C. Wsu, T. Kuo, W. Wu. 2009. Risk Evaluation of Green Components to Hazardous Substance Using FMEA and FAHP. Expert System with Applications 36, 7142-7147

Su, C.T., Chou C. J. 2008. A Systematic methodology for the creation of Six Sigma project: A case study of semiconductor foundry. Expert System with Application, 34 (4), 2693-2703

[18]

Xiang, W., L. Yunxian, Q. Zeliang. 2012. Estimation of Distribution: Implication Engineering Risk. System Procedia 3, pp. 132-138.

[5]

[6]

Jones, J.B.Jr. 2005. Hydroponics: A Practical Guide for Soilless Grower. CRC Press: New York, pp. 99

[7]

Kamble, V.S., T.Z. Quazi. 2014. FMEA of Shell Molding Process and Prioritizing by Using AHP. International Journal of Research in Aeronautical and Mechanical Engineering (2)6, 161-176

Zhenwei, S. Crop Yield for Crop Engineering

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Nanotechnology Use of Activated Carbon Coal to Decreasethe Paramters of Peat Water in Jambi Muhammad Naswir Education Department of Chemistry, Faculty of Teacher Training and Education, Jambi University, Email addres : [email protected] Abstract: This study aims to look at the characteristics of the activated carbon made from coal raw material, determining its ability to lower the water main peat parameters are: COD, color, organic matter, Fe, pH and TDS. Coal activated carbon made by pyrolysis with temperature 600-800oC, then activated by the addition of sulfuric acid compounds (HCl) 10% and acid phosphate (H3PO4) 10%. specifications of activated carbon is determined by measuring the moisture content, ash content, yield, number of iodine and methylene blue. Characterization and identification of activated carbon composition is done by using the instrument XRD and SEM-EDS. For the measurement of the parameters used mainly peat water TDS meter, pH meter, titrimetry methode, spectrophotometer UVVIS and AAS. The results showed that the activated carbon produced from coal has met the specifications set by the yield of 44% (standard 30%), the rate of Iod 998 mg / l (standard min 750 mg / l), methylene blue 225 mg / l (standard min 120 mg / l). Characterization by XRD showed that activated carbon coal contains carbon, silicate and quartz compounds, and heavy metals Cu and As. Activated carbon from coal has the ability to reduce the content of COD, color, Fe and organic substances, but has not been able to raise and lower the pH value of the water content of TDS in peat Keywords:Activated carbon, coal, XRD, SEM-EDS, water peat 1. Introduction Activated carbon is a porous material from the burning of materials containing carbon (C) with limited air, which has been done either by steam activation or by using a chemical activator Cheremisinof (2002). Activated carbon adsorbent largely used for various types of waste and fluids from such waste inudstri, factories, hospitals and so forth, which contain pollutants in the form of organic compounds and inorganic compounds, activated carbon is used as absorbent materials, cleaners or purifiers and also used as a catalyst role in small quantities by John et al, (2001). In the process of water purification, activated carbon can adsorb metals such as iron, copper and nickel. Besides activated carbon also serves to eliminate the odor, color and flavor contained in solution or waste water. Because charcoal is non-polar, then the nonpolar components of high molecular weight (4 to 20 carbon atoms) contained in the waste water plant can be adsorbed by activated charcoal by Ruey-Shin et al. (2002). The presence of activated carbon adsorption due to the micro pores very much, which can cause symptoms of capillary and surface area of activated carbon. Activated carbon is a molecule like a magnet that can form thousands of holes that can be used as the material forming the internal structure of the molecule. Activated carbon derived from a variety of materials in its manufacture can be through chemical and physical processes by using various methods. Some important factors in the process of characterization of activated carbon is to determine

the potential distribution absorptivity (APDs) and distribution of energy as well as the density. Density can be measured with a porosimeter by using the following equation: Aik Chong and, Jia Guo, 2001 and Vladimir and Gun'ko (2001) ε = [(ρs-ρa) / ρs] 100%. a = density of solids and density ρ The raw material for the manufacture of activated carbon is a material condition which contains containing carbon, such as; coal, coconut shells, palm shell, sawdust, bagasse and so forth. This research will be done manufacture of activated carbon using palm shells and coal material because the material deposit in many areas, especially in the island of Sumatra Indonesia, including Jambi region. There are three types of activated carbon forms, ie powder with a size of <0.18 mm, granular form with a size of 0.2 -5 mm and a pellet-shaped with a diameter of 0.8 to 5 mm. Coal is still rarely used as an activated carbon material, too much dekspor at low prices, but if made into activated carbon price is much greater than in exports in raw form. Krzystof (2000), give statement explains that coal is composed of carbon black and brownish precipitate formed from plants and organic materials that take place millions of years ago. Due to the influence of age, the pressure and heat, over millions of years to form coal seams of coal, which can be classified : into one group Lignite is brown coal, is rather soft and low organic maturity, are used as fuel in power plants. (2). Subbituminus is the result of dehydrogenation and methanogenesis from lignite.

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Have a maturity of higher organic, harder and darker than lignite. Used as fuel for power generation, cement production and use for industry. (3). Bituminous is further from the dehydrogenation reaction on the formation and separation of methane gas and hydrocarbon gas. Solid, black but sometimes dark brown. Other than as a fuel in power plants are also used for heat and power applications in the iron and steel industry and 4). Antracit is the last stage in which the process of coal formation is perfect. This is a type of coal with the highest quality and are widely used for space heating and settlements. Coal is a heterogeneous compound consisting of carbon-containing solids (C), hydrogen (H), oxygen (O), nitrogen (N) and sulfur (S). This substance is mixed with mineral, water, gas trapped in the pores. Due to the high carbon content of coal is possible to be processed into activated carbon, which can be used as an adsorbent for substances that are toxic, toxic gases and for clean water filtering materials Arfan (2006). Processing of coal into activated carbon can be done by heat activation at 700-1100oC or activation of acid with the addition of H3PO4 activator compound Cheremisinof, (2002), Marsh et al.(2006). Chemically activated carbon activation can be done with the addition of inorganic solvents such as, KOH, NaHCO3, NaOH, as has been done by Asyhar, 2005, Nguyen (2001) and Artur P. Terzyk et.al (2002). The use of activated carbon has been known to scientists are spacious enough, but the use of activated carbon as peat water treatment materials are still scarce done, in terms of peat water is relatively abundant and potential processed into clean water, which can be exploited by people who live in the area of peat which each while always clean water crisis. This research will be conducted experiments the use of activated carbon for water treatment peat into clean water. 2. Methodology 2.1 Equipment and Material The tools used in the study were; oven, furnace, filter 150 and 200 mess, analytical balance, stirrer (stirrer), filter paper, aluminum foil, sample bottles and glassware. Instrument used pH meter, TDS meter, visible spectrophotometry (UVVIS), Spektrometry Atomic Absorption (AAS), X-Ray diffraction (XRD), scanning electron Microscopoy (SEM) and Energy Dispertive Microscopy (EDS). The materials required include: coal, peat water, chemicals reagents HCl, H2SO4, buffer pH 4 and pH 10, HNO3, H2O2, K2Cr2O7, H3PO4, HgSO4, NH2SO3H, K2PtCl6, CoCl2.6H2O, and distilled water

2.2 Preparation of Activated Carbon Activated carbon in this study is made from coal and palm shell, activated carbon manufacturing process consists of pyrolysis feedstock followed by activation. Carbonization and heat activaion of coal carried out at a temperature of 300-600oC for 4 hours Activation is a process for eliminating carbon hydrocarbon that coats the surface of the carbon thereby increasing the porosity of charcoal. According to Said (2005) activator used is a compound H3PO4and HCl. Activation is done by soaking carbon from coal in 10% H3PO4 acid solution, soaked for 10 hours Teng (2000), then drained and washed with distilled water until the pH close to neutral, then dried in an oven at 170oC . Subsequently cooled. Activated carbon obtained in the specifications determined by determining the ash content, water content, the number of iodine and methylene blue numbers. Activated carbon is already ready tested for its ability to lower the color parameter, iron and organic matter of peat water. 2.3

Characterization of Activated Carbon

Activated carbon produced from coal determined specifications that yield, moisture content, ash content, the absorption of the iodine and methylene blue, then to see the shape and composition of the surface active compounds of carbon compouds performed using and SEM-EDS. X-RD the brand Philips Analytical PC-APD, Diffraction software Diffractometer type: PW1710 BASED, Tube anode: Cu Generator tension [kV]: 40, Generator current [mA]: 30 Wavelength Alpha1 [??]: 1.54060, Wavelength alpha2 : 1.54439, Intensity ratio (alpha2 / alpha1): 0.500, Divergence slit: 1 °, Receiving slit: 0:45 and Type of scan: CONTINUOUS and continued with test instruments SEM-EDS brand JEOL JSM 6510L A, which aims to see the surface structure and composition of elements and compounds constituent of the sample Zulkarnaen, et al. (1990). Air peat baseline test to determine the pH content, color, iron (Fe) and organic substances contained in peat water before being given the addition of activated carbon. Then put each of 500 ml water in a beaker glass peat, activated carbon is then added as much as 1, 2, 3, 4.5 g each of activated carbon, stirred for 5 minutes and precipitated for 2 hours, then filtered, and the filtrate was measured content COD, pH, Color, Iron (Fe), TDS, and organic substances 2.4 Measurement Parameter Water Peat The determination of the degree of acidity (pH) of water peat use potentiometric method, using a pH meter instrument, the normative reference (ASTM, (1982), measurements of organic substances using Permanganometri titration method, determination of

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the color of peat water is done by comparing the color of the test sample with a standard solution of color. In this study is used as a raw material solution of platinum cobalt color with PtCo scale unit. Quantitatively measuring the color using spectrophotometry instrument (SNI 06-2413-1991). The method used for determining the water content of Fe in the peat is method by using Atomic Absorbsion Spektrfotometry. Measurement of the peat content of COD in water before and after the treatment is done by using a titirmetri methode.From the measurement data carried doplo absorption efficiency is calculated by the equation

ń= efficiency of absorption (%), Co = initial concentration (mg / L) and Ce final concentration (mg / L) 3. Results and Discussion In this study, the raw materials used for the manufacture of activated carbon is; coal. The raw material coal in Muara Bungo regency of Jambi types of anthracite. Coal is collected in rough shape then reduced size, subsequent carbonization by burning at temperatures above 3000C. Then carbon pyrolysis results in the puree in nano size, so activated with asam clorida (HCl) and asam posphat (H3PO4). Activated carbon profile as shown in Figure 1.

row coal

CoalSEMCoal ctivated HCl

CoalSEMCoal not ctivated

CoalSEMCoal ctivated H3PO4

Figure 1.ProfileActivated carbon fromcoal (ASTM). Specifications coal activated carbon 3.1 Specificationactivated carboncoaland Palm produced is measured yield, content of water, ash, shell Activated carbon which has been produced from raw iodine numbers and the numbers of methylene blue. coal feedstock is determined by reference to the Results of the specification as listed in Table 1. standard specifications that have been defined Table1.Specificationsof activated carboncoalandpalm shell No Parameters Coal 1 Yield % 44 2 Water Content % 11,2 3 The Ash Conten % 2,4 4 Number Iod mg/g 998 5 Metylen blue mg/g 225 6 he paerticel sizxe 150 From Table 1 it is known that activated carbon produced from coal quality is good enough, where the average yield is 44%, the water content of 11.2%, while the absorption of iodine and

Standart 30 15 Max 10 Min 750 Min 120 Min 90 Methylene blue against a very good average of 998 ppm and 225 ppm. All parameters specifications in accordance with established standards.

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3.2 Analysisof carbonswitched onbyXray Diffraction Spectra of X-ray Diffraction of activated carbon coal and palm shell it can be seen in Figure 2

Figure 2. XRD spectra of activated carbon from coal and palm shell (as a comparison) Characterization by X-ray Diffraction in Figure 2 above shows that the activated carbon coal contains carbon and compound silicate and quartz, as it also activated carbon in the coal also include compounds silkat oxide (SiO2) and quartz and heavy metals such as copper (Cu) and arsenic (As). Activated carbon from coal is different in composition with activated carbon from palm shell materials. Activated carbon from palm shells XRD spectra show that that is not shaped (amorphous) and

composed of carbon compounds and 98.03%, 0.33% oxygen. 3.3 The results of SEM-EDS Analysis of Activated Carbon SEM-EDS analysis results for activated carbon coal and palm shell shows that the activated carbon have different content elements and compounds and compositions as set out in Table 2.

Table2.Content ofactivated carboncoalbeforeandafteractivationwithof H3PO4 No

Elemen

Unit

Non Activation

Activation HCl

Compound

Activation H3PO4

1 2 3 4 5 6 7 8 9 10 11 12 13

C O Al Si Cl P Ti Fe Cu As Cd Hg Pb

% massa % massa % massa % massa % massa % massa % massa % massa % massa % massa % massa % massa % massa

100 0 -

86,94 5,96 2,14 3,16 0,23 1,57 -

C Al2O3 SiO2 Cl TiO2 FeO CuO -

86,94 4,05 6,75 0,30 1,96

From Table 2 above are based on the SME-EDS spectrum can be concluded that the carbon content of coal are activated with H3PO4 has a content of 86.94% Al2O3 mass, 6.75% SiO2 compound. Then the iron content in the form FeO compound activated coal using H3PO4 was 1.24% mass, and coal contains no dangerous heavy metals (Cd, Hg, Pb and As), except for copper (Cu) by 1.96% by mass, in the form of CuO compounds. This fact is supported by the data SEM and EDS show that coal

has already been activated do not contain heavy metals. Photos of SEM-EDS, it is known that the number of pores is formed of activated carbon that has been activated oil cakang seems more and more open than before the activation, the activated carbon has a fine pore structure are many, activation greatly affects the surface area and total pore formed. Pore size average is formed is 0.336 lm.

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Figure 3.Foto SEM and spektra EDScoal activated with by using activatorH3PO4

Figure 4. Foto SEM and spektra EDS coal activated with by using activator HCl SEM photograph carbon active with 5000x magnification can be show surface shape and the pores are formed after and before it is activated, such as those in Figure 3. From the results of EDS is known that activated carbon from the oil that has not been activated cakang composed of the elements carbon 81.80% and 18.20% Oxygen. While those already activated carbon element content to be 100%, meaning that the carbonization process takes place perfectly. The

activation process is done greatly affect the quality and ability of activated carbon. Activation can be used to remove volatile matter and other gases (Bansal 1994). Activation can also dissolve metals heavy metals contained in coal. Later in the activation can open the pores of the activated carbon pre-existing but still closed, as can be seen in the picture above (Speinght, J.G. 1994 and Díaz-Terán (2001).

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From X-ray Diffraction Spektar as in Figure 4 below is a picture of peaks that appear, which shows the elemental composition and the constituent elements of each of the activated carbon 3.4 Decrease Parameters of Peat Water COD measurement results and the pH of the water content of the peat before being given the addition of activated carbon is 104 mg / l with a pH of 4.2. This condition is above the raw water. Clean water quality standards define the content of COD 10 Mg / L and pH 6-9, then the peat water can not be categorized as clean water or decent life. After being given the treatment of COD in water peat decreased to 73.5 mg / L. the decline was not too large with a pH of 3.9. Coal activated carbon's ability to reduce the water content of color on peat only 41.81%, organic matter 73.73%, Iron (Fe) 52.57%. And the addition of active carbon coal did not affect the significance of the pH and TDS in water peat, it can even degrade the water quality of peat can increase the acidity of the peat water into pH 3.54 to 4.82. The increased value of TDS in water peat after the addition of activated carbon due to the activated carbon in nano size is water soluble peat and form colloidal did not settle so that the solute in the peat water will rise. While the decline in water pH peat or increase in acidity caused by the activated carbon that is activated by the addition of a strong acid H3PO4, causing activated carbon is acidic (pH< 5), despite being washed with distilled water, but washing is not perfect and if activated carbon is mixed with water that is too acidic peat then the pH tends to decrease. The addition of H3PO4 activators can reproduce and enlarge the pores of activated carbon as described (Speinght 1994), that activation with KOH to form K2COO-, which led to the separation of carbon so as to increase porosity. The end of the pore structure and the development of surface area depending on the raw materials used and the history of carbonization (heating rate, final temperature, the atmosphere, the active agent, the ratio of activation). However, in addition to structural facts, adsorption can be greatly influenced by the presence of acidic or basic surface groups (carbonyl, phenols, lactones, quinones, etc.) in carbon. Thus the surface could change their non-polar character, making it more or less hydrophilic, change the point of zero charge and a negative surface charge density. According to Goyal (2001) and Díaz-Terán (2001) that there is a relationship between the porosity characteristics of the activated carbon with activated carbon energy and its ability to adsorb absorbent. In the carbonization process occurs water evaporation and decomposition of the components contained in the shell, namely cellulose,

hemicellulose and lignin. Singgih and Ratnawati (2010), hemicellulose decomposition process takes place at a temperature of 2000C to 2500C marked with thin white smoke during combustion. Cellulose decomposition process starts at a temperature of 2800C and ends at a temperature of 3000C to 3500C, this stage is characterized by the smoke thicker and darker, brownish black. Whereas lignin decomposition begins at a temperature between 3000C to 3500C and ends at temperatures between 4000C to 4500C in this process colors white smoke generated back and thin and disappear over time. In the carbonization stage, the raw materials and the air is heated without the addition of chemical substances. Carbonization goal is to eliminate the volatile matter and other gases present in coal Bansal (1998). Activation of carbon be done by chemical activation method acid activation that is by immersing the carbon with a strong acid (H3PO4), activation is an important process in the manufacture of activated carbon. Through the activation process, carbon will have a much better adsorption of carbon without activation. If not activated charcoal carbonization results still contain substances that cover the pores of the surface of the charcoal. At the time of soaking the activator solution will be adsorbed by the charcoal which will then dissolve mater-impurities that cover the pores of the charcoal. The loss of these substances on the surface of activated charcoal will cause the larger pores of activated carbon (Budiono, 2003). More and pore size of the activated charcoal result of the vast surface active surface so it will enhance the adsorption power of the activated charcoal. With the ability of activated carbon adsorbs various chemicals, the activated carbon usage continues to increase and expand, both for dealing with the waste industries, as a catalyst and as bleancing earth palm oil, and for the purification of water. Utilization of activated carbon filtration process for the production of drinking water has been developed by Heijman (1999) Activated carbon can reduce water parameters that meets drinking water standards, the same research on activated carbon has also been developed, activated carbon is used as support the process liquid-phase filtration material by AbdelNasser et al. (2001). Developed a combination of experiments on a mixture of calcium, silicates and phosphates directly and simultaneously to see the effectiveness of adsorption in the liquid phase. 4. Conclusion Activated carbon is made with raw materials Coal already meets the specifications of both the active carbon in accordance with the standard. Coal activated carbon is a type of mineral that is indicated by the sharp peaks in the chromatogram .Karbon

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XRD active coal has the ability to reduce the content of COD, color, organic matter and Fe in the peat water, but have not been able to raise the pH and reduce the content of TDS. From the results of this study can be recommended for future researchers in order to wash the activated carbon over and over again with distilled water until neutral pH, then to further increase the effectiveness of activated carbon in peat lowering water parameters necessary to study the combination with other materials such as with alkaline limestone. References [1] Cheremisinof, N.P. 2002. Handbook of Water and Wastewater Treatment Technology, by Butterworth Heinemann, USA. [2] John K. Brennan, Teresa J. Bandosz, Kendall T. Thomson, Keith E. Gubbins, .2001. Water in porous carbons, Colloids and Surfaces A: Physicochemical and Engineering Aspects 187188 (0), pp. 539 - 568. [3] Ruey-Shin Juang, Feng-Chin Wu, Ru-Ling Tseng, 2002. Characterization and use of activated carbons prepared from bagasses for liquid-phase adsorption, Colloids and Surfaces A: Physicochemical and Engineering Aspects 201 (1-3) (2002) pp. 191 – 199Aik Chong Lua, Jia Guo, 2001. Preparation and characterization of activated carbons from oil-palm stones for gas-phase adsorption, Colloids and Surfaces A: Physicochemical and Engineering Aspects 179 (2-3), pp. 151 - 162 [4] Vladimir M. Gun'ko, Duong D. Do, 2001. Characterisation of pore structure of carbon adsorbents using regularisation procedure, Colloids and Surfaces A: Physicochemical and Engineering Aspects 193 (1-3), pp. 71 - 83. [5] Krzystof Labus, 2000. Heavy-metal emissions from Coal Combustion in Southwestern Poland. Institute for Apllied Geology. University of Silesia, Poland [6] Arfan. 2006, Preparation of Coal-Based Activated Carbon with Controlled Activation Treatment And Performance Test. Depok. Department of Chemical Engineering FT-UI [7] Marsh, Harry, and Fransisco,R. 2006. Activated Carbon. London, Elsevier [8] Asyhar, R., Idiasari, R., Padliah, Melly, (2005c), Power Comparative Study of Adsorption Charcoal Shells Oil, Rice Husk and with Activator Activated Coal Chemistry, Journal of Chemistry, University of Edinburgh (the mold) [9] D.D. Do, C. Nguyen, H.D. Do, 2001. Characterization of micro-mesoporous carbon

media, Colloids and Surfaces A: P.and Engineering Aspects 187-188 (0), pp. 51 - 71. [10] Artur P. Terzyk, Piotr Kowalczyk, Piotr A. Gauden, Gerhard Rychlicki, Stefan Zitek, 2002. New relationships between the characteristic energy of adsorption and the average effective diameter of carbon slit-like micropores. Physicochemical and Engineering Aspects 201 (1-3), pp. 17 – 30 [11] Hendra, R. 2008. Preparation of Activated Carbon. Department of Chemical Engineering. FT-UI. Depok Indonesia [12] Said, N.I, 2005. Water Treatment Peat Simple. http: // www, kimprawil.go. id / Balitbang / Pusair / water [13] Teng. 2000. Influence of Different Chemical Reagents on the Preparation of Activation Carbon from Bitumininous Coal. Departement of Chemical Engeneering National Cheng Kung University. Taiwan [14] Zulkarnaen, Ward, S. Marble, DH 1990. Assessment Bentonite Processing and Utilization of the District Oule Kab. Terri East Java Province As Absorbent and Materials Drilling Mud. PPTM Bulletin Vol. 12 6 Jakarta P. 9-12 [17]. [15] ASTM, (1982), Standard Test Method for Volatile Matter in the Analysis of Coal and Coke, ASTM D-3175-3182 [16] Bansal, R.C. 1998. Activ carbon, New York. Marcel Dekker.Inc. [17] Speinght, J.G. 1994. Chemistry and Technology of Coal Second. Edition New York. Mercel Dekker. Inc [18] J. Díaz-Terán, D.M. Nevskaia, A.J. LópezPeinado, A. Jerez, 2001. Porosity and adsorption properties of an activated charcoal, Colloids and Surfaces A: Physicochemical and Engineering Aspects 187-188 (0), pp. 167 175. [19] Meenakshi Goyal, V.K. Rattan, Diksha Aggarwal, R.C. Bansal, 2001. Removal of copper from aqueous solutions by adsorption on activated carbons, Colloids and Surfaces A: Physicochemical and Engineering Aspects 190 (3) (2001) pp. 229 – 238 [20] Singgih Hartono and Ratnawati (2010). Manufacture of activated carbon from palm shells with chemical activasi method, Tangerang P 12-16, [21] Budiono A, et al, 2006. Effect of Activation Coconut Shell Charcoal with H2SO4 To Adsopsi Phenol. Undip

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[22] S.G.J. Heijman, R. Hopman.1999. Activated carbon filtration in drinking water production: model prediction and new concepts, Colloids and Surfaces A: Physicochemical and Engineering Aspects 151 (1-2) pp. 303-310 [23] Abdel-Nasser A. El-Hendawy, S.E. Samra, B.S. Girgis. 2001. Adsorption characteristics of activated carbons obtained from corncobs, Colloids and Surfaces A: Physicochemical and Engineering Aspects 180 (3) pp. 209 - 221.

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Utilization of Lignocelluloses from Agricultural Waste as Raw Material for Producing Bioethanol Sriharti, Wawan Agustina, Takiyah Salim dan Lia Ratnawati Center of Appropriate Technology Development – Indonesian Institute of Sciences Jl. KS. Tubun No. 5 Subang – West Java – Indonesia Abstract: Indonesia needs to develop bioethanol by reasons of ever increasing energy consumption, increasingly diminishing stock of oil fuels, increasing oil fuel imports. A conversion of lignocelluloses into bioethanol can be devised to substitute gasoline fuel for transportation purposes. The agricultural wastes-derived lignocelluloses materials are available abundantly and not utilized as food stuffs, and thus their utilization as a source of energy will not interfere with food supply. Lignocelluloses are composed of three main components, namely, cellulose, hemicelluloses, and lignin. A conversion of lignocelluloses materials into ethanol consists basically of a pretreatment, saccharification/hydrolysis of cellulose and lignocelluloses into sugar, fermentation of sugar into ethanol, and refinement of ethanol by distillation and dehydration processes. Currently, the production cost of ethanol by using lignocelluloses is still quite high, particularly for the pretreatment. Therefore, a study is required to improve the productive processes from pretreatment to ethanol refinement. The study findings revealed that the biologic pretreatment by using cellulose enzyme and lignocelluloses-producing local microbes was the best process, being more environmentally friendly and of lower production cost. Keywords: agricultural wastes, lignocelluloses, conversion, bioethanol 1. Introduction Currently, oil fuel (BBM) has been a problem due to the increasingly diminishing stock of fossil fuel and at the same time the increasing rate of its consumption. As a result, the government has cut BBM subside, taken some measures of saving energy, and been seeking alternative energy sources to substitute for earth oil. The government issued a Presidential Regulation No. 5 of 2006 on National Oil Policy, where the utilization of biofuel in targeted by 2% and 5% in 2010 and 2025, respectively (Tim Nasional Pengembangan BBN, 2008). Bioethanol may be utilized as fuel substituting for gasoline and as a premium mixture material. Its utilization as fuel espouses the world conference in searching for a second generation fuel alternative, i.e., a fuel derived from a mixture of earth oil fuel and renewable fuel (Mulyana, Y., 2008). Bioethanol is one of our future energies. An Organization for Economic Cooperation and Development (OECD)‟s report estimated that 13 per cent of liquid fuel shall be needed by 2050, supplied from transportation bioethanol. The estimation is based on both technological and economic potentials of bioethanol. Transportation bioethanol cannot yet replace fossil fuel totally in near future; instead, it will be only a partially alternative solution (Doornboch and Steenblik, 2007). Bioethanol is an environmentally friendly fuel with a high oxygen content (35 per cent) and thus much cleaner when burned. Moreover, its carbon monoxide emission is lower by 19-25 per cent below BBM, and therefore it doesn‟t contribute to the accumulation of carbon dioxide in atmosphere and is renewable (LIPI, 2011).

Indonesia is potential to be the biggest bioethanol producer in the world, because the raw materials of bioethanol are plants abundantly growing in Indonesia, that is, ones that contain starch and sugar, such as cassava, corn, sugar cane, sugar palm, sweet potato, and sago palm. Based on the raw materials used, bioethanol is distinguished into four generations. First generation utilizes sugar- or starch-based raw materials. This fermentation process produces an ethanol by 12 per cent in maximal. Second generation utilizes raw materials containing lignocelluloses derived from agricultural, forestry, or plantation and industrial wastes. Third generation utilizes sea algae raw material. And fourth generation utilizes the raw materials derived from a biomass that has already been modified genetically so as to accelerate the bioethanol pretreatment processes. The lignocelluloses-containing biomass is derived from agricultural wastes, plantation wastes, forestry wastes, solid paper wastes, and some industrial wastes. Lignocelluloses biomass is a remarkable substrate as an alternative bioethanol because it doesn‟t compete against food stuffs and of cheaper prices (Samsuri, 2007). In developing the technology of a bioconversion of biomass wastes into bioethanol, at a commercial scale, a pretreatment of lignocelluloses wastes has to be done such that it results in a significant product, given that a pretreatment is a stage that is both costly and of reasonable effect on the cost of the entire process. The high production cost is particularly resulted from the low work of enzyme in the substrate due to both cellulose crystallization and the

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presence of inhibitors that may decrease the fermentation of cellulose and hemicelluloses into ethanol (Stenberg et al., 1998). Therefore, to optimally produce bioethanol from lignocelluloses, an optimization of productive processes, particularly in pretreatment, hydrolysis (saccharification), and fermentation processes, is needed. The objective of the present research was to investigate the utilization of lignocelluloses from agricultural wastes. The research discussion was focused on the characteristics of lignocelluloses, the technology of the conversion of lignocelluloses into ethanol, beginning from pretreatment, saccharification, fermentation of sugar into ethanol, to ethanol refinement. 2. Characteristics of Lignocelluloses The main content of lignocelluloses material is composed of three different types of polymer, namely, lignin, hemicelluloses, and cellulose, mutually bounded to form an integrated unity (Perez et al., 2002). The content of each component depends on the type of biomass, age, and environmental condition of the location where the biomass is growing and developing. According to Schacht et al., (2008), the characterization of the lignocelluloses of biomass consists of cellulose, hemicelluloses, and lignin by 35 – 50 %, 10 – 25 %, 10 – 25 %, respectively. Cellulose of (C6H10O5)n is a polysaccharide composed of D-glucose connected uniformly by βglucosidal bond. Its polymerization degrees are designated by n, with a broader approximation varying from thousands to tens of thousands. This form of polymer enables the celluloses to be interconnected/bounded into a very strong fiber. A total hydrolysis of cellulose produces D-glucose (a monosaccharide), whereas a partial hydrolysis produces a disaccharide (selobiose) and a polysaccharide with successive ns from 3 to 10. Cellulose has crystal structures and of high resistance to both acid and base (Badger, P.C., 2002). Moreover, hemicelluloses are a polysaccharide the units of which consist of 5carbon monosaccharide such as D-xilosa and D-arabinosa and 6-carbon monosaccharide like G-manosa, D-Galactose, and D-glucose. The 5-carbon monosaccharide outnumbers the 8-carbon monosaccharide with an average molecule formula of (C5H2O4)n. Because the polymerization degree (n) of hemicelluloses varies from 50 to 200, less than that for cellulose, it is easier to break than cellulose and most hemicelluloses are alkali soluble in nature. Common hemicelluloses are xilam, consisting of xiloma of a 1,4,glucomanan bond. The enzymes that fall into hemicelluloses components are, among others, xylanase, βmanannase,α-L-arabino-furanosidase, α-

D-glucuronidase, β-xylosidasedan hemicellulotic estarase (Shallom and Shoham, 2003). Hemicelluloses are abundantly produced by Aspergillus and Trichoderma molds (Gerhartz, 1990). 3. Potentials of lignocelluloses-based agricultural wastes in Indonesia The development and advancement of agriculture and agricultural industry in Indonesia have resulted in increases in agricultural wastes, most of which are lignocelluloses-containing wastes. Basically, wastes may produce environmental pollution with no economic value, even of negative value because they incur some treatment cost. Lignocellulosescontaining wastes as an organic material are of great potential to be utilized as a raw material in bioethanol production. Selecting the type of material should consider its supply. The table below presents the potentials of agricultural wastes that contain lignocelluloses. Table 1: The potentials of lignocelluloses-based agricultural wastes in Indonesia Product/c ommodity Paddy Rice Cassava

Agricultural Potentials Wastes Rice stalks 5,000 kg/ton paddies Rice husk 280 kg/ton paddies Stem/stalk 800 kg/ton cassava Heaps/tapioca dregs Corn Corn hump 700 kg/ton corn and husk Coconut Fiber 280 kg/ton coconut Shell 150 kg/ton Cane Bagasse 280 kg/ton Oil palm Fiber and shell 420 kg/ton CPO Empty stem 200 kg/ton fresh stem Source: Falah, F. (2012)

4. Technology of Conversing Lignocelluloses into Ethanol To reproduce bioethanol from agricultural wastes that contain lignocelluloses, according to Howard et al., (2003), there are 4 stages, namely: 1) physical, chemical, physico-chemical, and biological pretreatment, 2). Polymer saccharification/hydrolysis (cellulose, hemicelluloses, lignin) into plain sugar (hexosa, xylosa), 3) Sugar fermentation by microbe to produce ethanol, and 4). Separation and refinement of ethanol produced. 4.1 Lignocelluloses Pretreatment Pretreatment is a costly stage and significantly affects the cost of the whole process. The purpose of a pretreatment is to open lignocelluloses structure for cellulose to be more accessible to the enzymes that breaks polysaccharide polymer into glucose (fermentation sugar).

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There are some lignocelluloses technologies, among others:

pretreatment

 Physical pretreatment in form of mechanical chopping, grinding and powdering, to reduce the size of material and cellulose crystallinity, and to increase the width of surface in contact with enzymes, needs large energy (Sun and Cheng, 2005).  Physico-chemical pretreatment: a combination of physical pretreatment and chemical pretreatment of among others vapor explosion, liquid hot water, CI2 explosion and ammonia fiber explosion (AFEX). By the method, biomass particles are exposed to both high temperature and pressure and then the pressure is rapidly decreased, so that the material undergoes an explosive decompression, decreasing cellulose chrystallization (Ballesteros, et al., 2006, Li et al., 2010).  Chemical Pretreatment: addition of chemical compound, such as acid hydrolysis, alkali hydrolysis (decreasing polymerization degree), ozonolysis, oxidative delignification), and organosolvent process (Saha, et al., 2006; Dien et al., 2006).  Biologic pretreatment, using enzyme-producing microorganisms, such as white weathering mold microorganism (Pleurotus ostreatus), brown weathering mold, soft weathering mold to degrade lignin and hemicelluloses (Tengerdy and Szakacs, 2005). Biological method is more environmentally friendly, consuming very low energy, and fairly effective in degrading lignin. Because of the diversity of lignocelluloses materials, different raw materials need different pretreatments. Therefore, there is no one general method applies to the pretreatment of all lignocelluloses materials. 4.2 Saccharification Saccharification or hydrolysis is a process of breaking down cellulose into selobiose and then into plain sugars like glucose. Hydrolysis is a chemical reaction breaking water molecules (H2O) into hydrogen cation (H+) and anion hydroxide (OH-) by a chemical process. The process is usually applied to break the organic polymer with carbon chains. There are three widely used hydrolysis methods, namely : a. Dilute acid hydrolysis. The use of dilute acid in a cellulose hydrolysis process is carried out at a high temperature in a short reaction time (in minutes). The temperature required achieves 200 o C; the dilute acid used is 0.2 – 4 % (Nguyen and Tucker, 2020). The use of dilute acid to hydrolyze cellulose may achieve a conversion of reaction up to 50 % (Badger, 2002). Some widely

used acids include sulphate acid (H2SO4), percolate acid, and HCl. Sulphate acid is one used most in acid hydrolyses. The disadvantages of the process are that cellulose and acid react fast and continuously. The reaction not only transforms cellulose into sugar, but also breaks down the formed sugar into other simpler compounds such as furfular. The breaking down of sugar reduces the produced sugar and inhibits the fermentation of sugar into ethanol, and thus the compound should be separated before the sugar fermentation, meaning more production costs. b. Concentrated acid hydrolysis. The use of a concentrated acid in a cellulose hydrolysis process is performed at lower temperatures, i.e., 100oC and needs a reaction time of 2 – 6 hours at acid concentration of 10 – 30 % (Zimbardi et l., 2002). The advantage of using a concentrated acid is a high conversion of sugar produced, that may achieve 90 % (Badger, 2002). The disadvantage of the reaction is that the reaction time needed is longer and needs a good washing process to reach the reaction pH before microbes are added in fermentation process. c. Enzyme hydrolysis utilizes either cellulose and hemicellulose enzymes or celluloseproducing microbe. An enzymetically hydrolysis process is usually proceeding under a light condition, pH around 4.8, and at temperature of 45 – 50 oC. The three main enzymes that exist in complex cellulose are endogluconase, exogluconase, and selobiase (β-glucosidase). Endogluconase hydrolyzes 1.4-β-glicocydic bond randomly in cellulose amorphous area to produce glucose, selubiose, and selodextrin. Exoglucanase hydrolyzes selodextrin by disconnecting selubiose unit from the end of polymer chain. And selobiose hydrolyzes both selubiose and selo-oligosaccharide into glucose (Sun, Y.,, Cheng, J., 2002). Hemicelluloses are a group of enzymes capable of hydrolyzing hemicelluloses. The hydrolysis of hemicelluloses can be monitored by the numbers of D-xilosa, Larabinosa, D-glucosa, D-mannosa, D-galactose, and L-amnosa produced. Given its capacity of hydrolyzing xilan, hemicelluloses are also called xinalase. Trichoderma viride is one of the microorganisms capable of producing cellulase enzyme to break cellulose into plain sugars. Trichoderma viride falls into Trichoderma genus, Moniliceae family, and Moniliales orde. The mold is easily visible due to its cotton-like fibrous appearance, but when its spores have emerged then it will be dark green in color (Domsch and Gams, 1972). Trichoderma viride is capable of producing complete cellulase enzyme complexes, i.e., endocellulase and

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exocellulase, capable of hydrolyzing crystalline cellulose and non-crystalline cellulose. In cellulose enzyme, a synergy between endocellulose and exocellulose, and also among execellulases, can take place. The lignocelluloses structure, composed of the cellulose and lignin matrix bounded by hemicelluloses chains, has to be broken to be easier to attack by enzymes during a hydrolysis process (Laureano-Perez et al., 2005). The factors that affect the capacity of enzymes in hydrolyzing lignocelluloses materials are among others lignin and hemicelluloses contents and cellulose crystalline level. Therefore, a pretreatment is needed to (1) remove lignin and lignocelluloses, (2) decrease cellulose crystalline level to increase cellulose amorphous fraction, and (3) increase the porosity of materials (Hsu et al., 2010). The pretreatment should also be capable of enhancing the capacity of forming sugar during a hydrolysis process, inhibiting the formation of inhibitor in next hydrolysis and during fermentation process, inhibiting the removal of carbohydrate, and being cost effective (Su, Y. and Cheng, J., 2005). An advantage of enzymatic hydrolysis is its highly efficient reaction, because the enzyme is selective and the formation of by-product can be eliminated and the temperature and pressure of reaction are low, even it can be conducted at room temperature and atmosphere pressure. Meanwhile the disadvantages of an enzymatic hydrolysis process is its longer reaction times that may achieve 72 hours (Kandono, B., 2010), and somewhat expensive prices of enzymes. The component of enzyme cost may reach 53 – 65 % of the cost of chemicals, the latter being around 30 % of the total cost. 4.3 Fermentation In a fermentation process, the formed plain sugars are then fermented into ethanol by aid of microbes such as leavened (khamir) Saccharomyces cerevisiae and bacteria Zymmonas mobilis. Fermentation is usually carried out at 30 oC, pH 5, and slightly aerobic. In a process of glucose fermentation, one molecule of glucose produces two molecules of ethanol and two carbon dioxides (CO2), by a reaction as follows: C6H12O6 + S. cerevisae2C2H5OH + CO2 + 2 ATP Glucose

yeast

ethanol,carbon dioxide, energy

The fermentation of the results of a hydrolysis of hemicelluloses component such as xilosa into ethanol may use a leavened Pitchia stipitis or Candida shehatae (Hahn-Hagerdal et al., 1993). In the fermentation of xilosa, three xilosa molecules produce five ethanol molecules, five CO2 molecules, and five water molecules (Milan, Mc., 1993 in Hermiati, et al, 2010). The fermentation of pentose

derived from hemicelluloses is conducted in a separate reactor because the microbes that use pentose work more slowly in converting both hexose and pentose into ethanol than those microbes that convert only hexone into ethanol, and more sensitive to both inhibiting compound and ethanol products (Cardona , C.A. and Sanchez, O.J., 2007). 4.4 Simultaneous Saccharification Fermentation (SFS)

and

In general, a synthesis of lignocelluloses wastesderived bioethanol consists of two phases, i.e., hydrolysis and fermentation separately conducted. The latest method is a simultaneous sacchrafication and fermentation (SFS) process, that is, a combination between enzyme hydrolysis and fermentation performed in one reactor. The process has an advantage, that is, the polysaccharide converted into monosaccharide will immediately be fermented into ethanol (Samsuri et al., 2007). The microbes used in a SFS process are usually cellulase enzyme-producing molds, such as Trichoderma reesei, Trichoderma viride, and leavened Saccharomyces cerevisiae. The optimal temperature of an SFS process is 38 oC, a combination of the optimal temperature of hydrolysis (45 – 50 oC) and that of fermentation (30 oC).

A SFS process has some advantages relative to instages hydrolysis and fermentation processes. The advantages of SFS according to Sun, Y., and Cheng, J. (2002) are:  Increasing the velocity of hydrolysis by conversing the sugar formed from the result of cellulosa hydrolysis that inhibits the activities of cellulase enzyme.  Reducing the needs of enzyme.  Increasing product rendemen.  Reducing the needs of sterile condition because glucose in immediately converted into ethanol.  Time process is shorter.  The volume of reactor is smaller because only one reactor is used. A SSF process is more tolerant to inhibitor compounds such as acetate acid that is formed by or derived from a pretreatment process existing in a liquid fraction. Therefore, in a SSF process, the pretreatment result materials can be immediately processed with no need to separate in advance the liquid fraction from solid fraction. The researches to develop in relation to the conversion of lignocelluloses materials into ethanol, according to Grohmann, 1993, are as follows:  Selection of lignocelluloses materials and their pretreatment.

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 Selection and development of microbes or enzymes for a hydrolysis process.  Selection and development of microorganism for fermentation a process.  Adaptation and development of an analysis technique to monitor the benefits of SSF. One of the important factors in selecting lignocelluloses materials to be converted into ethanol is a cellulose-to-lignin ratio. To obtain a high rendemen, the content of cellulose must be high and that of lignin low. In fermentation processes, heat- and ethanol-tolerant leavened, a selobiosafermenting leavened, and sugar-fermenting microbes derived from heicellulose, e.g., xylose, arabinose, galactose, and mannose, should be developed. 4.5 Distillation The content of ethanol produced by a fermentation process usually only achieves 8 – 10 %. An ethanolrefining process can be conducted by a distillation. Distillation is a process of evaporation and recondensation, separating a mixture of two or more liquids into their fractions according to the differences in boiling points that can be performed at 80 oC, as the boiling points of alcohol and water are 78 oC and 100 oC respectively (Miska, Y., et al., 2013). Distillation is a method of separating chemicals according to differences in either velocities or volatilities of materials. In a distillation, the mixture of substances is boiled until evaporating, and then the vapor is cooled down again into a liquid form. The substance with a lower boiling point will evaporate earlier. The application of the process method is based on a theory that in a solution each component will evaporate at its boiling point. The maximum concentration of ethanol that can be gained by a common distillation is 95 %. A bioethanol with a concentration of 95 % cannot be used as fuel yet. 4.6 Dehydration The bioethanol used as a fuel mixture for vehicles must be absolutely dry and anhydrous to ensure that it is non-corrosive, and hence it should be 99.5 % in grade. Therefore, a dehydration process is needed. A dehydration process is performed to remove water in an azeotrophic ethanol mixture (95-96 % ethanol), comprising 5 stages. First stage is a so-called azeotrophic process, conducted by adding either benzene or cyclohexane into the mixture. It will form a heterogeneous azaotrophic mixture. The final results are an anhydrate ethanol and a mixture of water and cyclohexabe / benzene. When condensed, the vapor turns into liquid. Other method is an extractive distillation, that is, by adding a terner component into hydrate ethanol, so that it will increase the relative instability of the ethanol. When

the terner mixture is distilled, it will produce anhydrate ethanol. 5.

Conclusion

 The production of bioethanol from lignocelluloses materials is a 4-stages process, consisting of pretreatment, saccharification, fermentation, and ethanol distillation.  Enzymatic hydrolysis process by using cellulose enzyme- and hemicellulosesproducing local microbes is the best one, being environmentally friendly, reducing the fermentability of cellulose and hemicelluloses into ethanol, and of lower production cost.  Both production and application of bioethanol from lignocelluloses materials are still faced with some barriers and constraints, among others, not fully mastered technologies, still low bioethanol rendemen, and still high prices of lignocellulosesderived bioethanol, and thus it is not competitive against the prices of oil fuel the government is still subsiding. Therefore, the role of government in subsiding lignocellulosesderived ethanol prices is required. Reference [1] Ballesteros, M., I., Negro, M.J., Olivia, M.J.M. Cabanas, A., Manzanares, P. and Ballesteros, M., 2006. Ethanol Production from Steam Explotion Pretreated Wheat Straw, Applied Biochemistry, Biotechnology, 70 – 72, 3 – 15. [2] Badger, P.C., 2002. Ethanol from Cellulose, A General Review, p. 17 – 21, In Janick and Whipkey, A Trends in New Cropnad New Uses, ASSH Press Alexandria, VA. [3] Cardona, C.A. and Sanchez, O.J., 2007. Fuel Ethanol Production: Process Design Trends and Integradation Opportunities, Bioresourche Technology, 98: 2415 – 2457. [4] Dien, B.S., Jung, H.J.G., Vogel, K.P., Casler, M.D., Lamb, J.F.S., Iten, L., Mitchell, R.B., and Sarah, G., 2006. Chemical Composition and Response to Dilute –acid Pretreatment and Enzymatic Sachararification of Alfafa, Reed Canarygrass and Switchgrass, Biomass Energy, 30, 880 – 891. [5] Doornbosch, R., and Steenbilk, R. 2007. Biofuels in the cure worse than the disease, OECD, Report. [6] Domsch, K.H. and Gams, W., 1972. “Fungi in Agricultural Soils. London: Longman Group Limited Publishing. [7] Falah, F., 2012. Pemanfaatan limbah Lignin dari Proses Pembuatan Bioetanol dari TKKS sebagai Bahan Aditif pada Mortar, Tesis, Fakultas Teknik Universitas Indonesia, Depok.

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[8] Gerharz, W., 1990. Enzyme in Industry: Production and Application, VCH Verlagsesellschaaft mbH, D 6940 Weinheim, p. 81 – 82. [9] Han Hagerdal, B., Hallborn, J., Jeppason, L., Olsson, L., Skoog, K. And Zacchi, G., 2006. Bioethanol the fuel of tomorrow from the residue of today, Trend in Biotechnology, 24 (12) : 549 – 556. [10] Hermiati, E., Mangunwidjaja, d., Sumnarti, T.C., Suparno, O., dan preasetya, B., 2010. Pemanfaatan Biomassa Lignoselulosa Ampas Tebu Untuk Produksi Bioetanol, Jurnal Litbang Pertanian, 29 (4). [11] Hsu. T.C., Guo, G.L., Chen, W.H. and Hwang, W.S., 2010. Effect of Dilute Acid Pretreatment of Rice Straw on Structural Properties and Enzymatic Hydrlolisys, Biosource Technology, 101, 4907 – 4913. [12] Laureano Perez, Teymouri, F., Alizadeh, H., and Dale, B.E., 2005. Understanding Factors That Limit Enzymatic Hydrolysis of Biomass, Applied Biochem, Biotechnol, 121 -124, 1081 – 1100. [13] Li, B.Z., Balan, V., Yuan, Y.J. and Dale, B.E., 2010. Process Optimization to Convert Forage and Sweet Bagasse to Ethanol Based on Ammonia Fiber Expansion (AFEX) Pretreatment, Biosource Technology, 101, 1285 – 1292. [14] LIPI, 2011. LIPI Kembangkan Bioetanol Generasi Kedua, Teknologi Terbaru Informasi Tentang Teknologi Terbaru. [15] Kardono, L.B., 2010. Teknologi Pembuatan Etanol Berbasis Lignoselulosa Tumbuhan Tropis untuk Produksi Biogasoline, Pusat Penelitian Kimia, LIPI. [16] Miska, Y., Ratnasari, D., Budi, N., Haerani, Usman, N., 2013. Teknologi Bioproses Bioetanol, Jurusan Teknik Kimia Politeknik Negeri Ujung Pandang. [17] Mulyana, Y., 2008. BBM Genersi Kedua, http://bioethanol indonesia.blogspot.com/ 2008/01/bbm-generasi-kedua.html. [18] Nguyen, Q.A. and Tucker, M.P., 2002. Dilute Acid / Metal Salt Hydrolysis of Lignocelulosic, United State Patent 6423145.

Biological Treatments of Cellulose, Hemicellulose and Lignin: An Averview, int. Microbiol, 5, 53 – 63. [21] Schacht, C., Zetzl, C. and Brunner, G., 2008. From Plant Materials to Ethanol by Means of Supercritical Fluid Technology, The Journal of Supercritical Fluid, 46, 299 – 321. [22] Saha, B.C., Iten, L.B., Cotta, A., Wu, Y.H., 2005, Dilute Acid Pretreatment Enzymatic Saccharirification and Fermentation of of Rice Hulls to Etanol, Biotechnology Progress, 2, 816 – 822. [23] Shallom, D., and Shoham, Y., 2003, Microbial Hemicellulases, Current Opinion in Microbiology, 6: 219 – 228. [24] Samsuri, M.M., Gozan, R., Mardia, M., Baiquni, Hermansyah, Wijanarko, Prasetya, Nasikin, 2007. Pemanfaatan Selulosa Bagas Untuk Produksi Bioetanol Melalui Sakarifikasi dan Fermentasi Serentak dengan Enzim Xilanase, Makara Teknologi 11; 17 – 24. [25] Stenberg, K., Tengborg, C., Gable, M. And Zacchi, C., 1998. Optimisation of Steam Pretreatment of SO2 Impregneted Mixed Softwood for Ethanol for Ethanol Production: Journal Chem. Technol. Bioethanol, 71 : 299 – 308. [26] Sun, Y., Cheng, J., 2002. Hydrolysis of Lignocellulose Materials for Ethanol Production: A Review Biosource Technology 83: 1 -11. [27] Tengerdy, R.P. and Szakacs, G., 2003. Bioconversion of Lignocellulose in Solid Substract Fermentation, Biochemical Engineering Journal, 13, 169 - 179. [28] Tim Nasional Pengembangan BBN, 2008. “BBN Bahan Bakar Nabati: Bahan Bakar Alternative dari Tumbuhan sebagai Pengganti Minyak Bumi dan Gas”, Penebar Swadaya, Jakarta, pp.164. [29] Zimbardi, F., Viola, E., Gallifuoco, A., Cantarella, M., Barisano, D., Braccio, G., “Overview of Bioethanol Production”, Universita degli Studi de L‟Aquilla, Dipartimento di Ingegneria Chimica e dei Materiali, Monteluco di Rojo.

[19] Palonen, H., 2004, “Role of Lignin in the Enzymatic Hydrolysis of Lignocellulose”, VIT Biotechnology, Helsinki University of Technology, Finland. [20] Perez, J., Munoz-Dorado, J., Rubia, T. and Martinez, J., 2002. Biodegradation and 1st International Conference on Appropriate Technology Development 2015, Bandung, October 5th-7th, 2015 | 164

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Influence of Local Wisdom to Prevent Disappearance of Cebong Lake in Sembungan Village Wonosobo District Putri Nurfahmia, Rizal Faozi Malika, Ratih Paniti Saria, Afid Nurkholisa a)

Gadjah Mada University, Bulaksumur, Yogyakarta 55281, Indonesia Abstract: Dieng Plateau which is located in Wonosobo district is has more than 2000 masl region. Lake in Dieng Plateau were formed by volcano activities, include Cebong Lake. This lake is located under Pakuwojo, Saroja and Sikunir foot mount. Sedimentation is eminent problem in Cebong Lake, because it may silting up and repeal the lake. Disappearance of Cebong Lake would threaten any human activities whose trade on its resources especially for farming activities as the main activity in this region. In addition, inhabitants want to preserve Cebong Lake. Therefore, they have to be a wise to environment. Methods used in this research are direct measurement of bathymetry mapping using echosounder, interpretation of suspended sediment in Cebong Lake, and interview. Result from this research is the lifespan prediction of Cebong Lake to supply water resources for farmers. In addition, it is concluded that local wisdom re-actualization is important to gain longer life span of the lake and impede its disappearance. Keywords: local wisdom; Cebong lakes lifespan; bathymetry; sedimentation. 1. Introduction Lake in Dieng Plateau ware formed by volcano activities, include Cebong Lake. This lake is located in Sembungan village Wonosobo district, Central Java, Indonesia. This lake located under Pakuwojo, Saroja and Sikunir foot mount. The characteristic of this lake is eutrophic lake. Land use in the lake environment (Pakuwojo, Saroja and Sikunir mount) is agriculture land with potato as the main vegetation. Agriculture develop rapidly in Dieng Plateau. Because of that, deforestation be something that can‟t be reined. The land conversion caused the land erosion intensively, especially at three decade ago (Setiawan, 2012). This condition become special attention for limnologist, because precipitation in Dieng Plateau is high up to 3917 mm/year (Andiana, 2007). It may cause big land erosion and enter the lake due to the land use. Dieng has a high rate erosion up to 9,2 kg/m2 at the potato agriculture area (Setiawan, 2012). Sedimentation is eminent problem in Cebong Lake, because it may silting up and repeal the lake. Catchment area around Cebong Lake has slope up to 30-40% with potato agriculture land use. Potato agriculture area located around Cebong Lake has contribution to increase erosion rate. Run off from catchment area take the sediment along and it‟s accumulated in the lower land, it is Cebong Lake. Sedimentation may influence any important thing in the Cebong Lake ecosystem. Decreasing of Cebong Lake‟s life span is one consequence of the sediment accumulation so it may decrease the water capacity in the lake. This sediment accumulation also may cause the disappearance of Cebong Lake. In addition, sedimentation also has side-effect for water quality in the Cebong Lake like Dissolve Oksigen

(DO) or any chemical content in the water of Cebong Lake due to fertilizer utilizing. Fertilizer utilizing also may cause eutrophication, over fertility. Water ecosystem in Cebong Lake may disturbed. The example is decreasing of fish population in Cebong Lake, its mean that water quality in the Cebong Lake is decrease. The detriment of physical environment is related with human behavior. This detriment can be prevented by human empowerment on local wisdom. The value of togetherness in local wisdom regarding human behavior with others include withthe environment (Santosa, 2011). The balance of natural resources usage and conservation produce the sustainable development. Actualization of local wisdom keep natural resources to be useful not only for present but also for the future. The purpose of this paper is to find out the local wisdom of Sembungan village to carry out Cebong Lake. This paper also discuss about physical characteristic of Cebong Lake. 2. Method and Theory 2.1 Lake and Sediment Lake/ pond/ reservoir can be defined as an enclosed body of water surrounded by land and has no access to the sea (Thomas et al, 1992). Lake has many functions, among others as a raw source of drinking water, irrigation, power generation (hydropower), fisheries and so forth, so that the lake plays an important role in the economic development of the region (Pusat Litbang Sumberdaya Alam, 2012). According Suyono (2004), a body of water can be referred as a lake if : 1) deep enough to form a vertical strata of water temperature;

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2) the floating vegetation is not sufficient to cover the entire surface of the water and just cover the edge of the lake; 3) it showed waves that could form the barren or wave swept shore. Sediment is the result of various erosion process, either because of cliff erosion, soil erosion, or other erosion. That erosion then transported and flows into the lower place to the lake by surface flow (Manalu, 2012). Sediments in Cebong Lake can be rotten by potato crop and plants that died and decomposed by bacteria that live in the Cebong Lake. 2.2 Local Wisdom Local Wisdom is any understanding, confidence, knowledge, insight, customs, and ethics as the reference, as well guideline guiding humans behave in life at ecological community. Cultural landscape is a real appearance of the interaction, adaptation, and adjustment of the human with the natural environment (Santosa, 2011). Different physical environments would produce different social conditions. 2.3 Method Methods used in this research are direct measurement of bathymetry mapping using echosounder, interpretation of suspended sediment in Cebong Lake, laboratory analysis and interview. Echo-sounder is an instrument for measuring the depth of water or a tool for sounding activities. This tool can measure the water depth from the surface of the water until the water base. Results from the echo-sounder is the depth point that is processed into bathymetric map (Bruinsma, 1997). Advantages of echo-sounder is able to record continuously from one point to another point. Echo-sounder can distinguish the materials of mud, clay, or sand and the layer thickness can be estimated. To predict the remaining life of a lake can be calculated by the amount of the average entry of sediment into the lake minus sediments output in one year, then multiplied by the number of years of lake operation (Nursa‟ban, 2008). The results of these calculations can obtain lifespan prediction of the lake. Measurements of sediment in Cebong Lake can use the bathymetric data with temporal measurement so it looks the addition of sediment each year and can predict capacities of Cebong Lake. Laboratory analyzes performed to determine the substances that contained in the lake. Parameters used are pH, DHL (electrical conductivity), TSS (Total Suspended Solid) and TDS (Total Dissolve Solid). The last method is interview. The interviews were conducted using depth interviews individually.

3. Social Characteristic Sembungan village has a socio-economic uniformity. That uniformity form of livelihood which produce similarity of needs. The main requirement of agrarian society in Sembungan village is irrigation water. Irrigation water supply in Sembungan agricultural areas comes from two sources. Rain as a source of irrigation water in the rainy season and the lake as a source in the dry season. Cebong Lake is a natural resource which is used together. Based on interviews with several people in the Sembungan village, can be described some information related to the preservation of Cebong Lake. That information is about tradition are still maintained by people in Sembungan village, such as cleaning the Cebong Lake during the Sya'ban month, nyadran, shaving gimbal kids, mujahadah, and marawis. In addition, there are some rules related to the treatment in Cebong Lake, such as people are not allowed to swim in the lake, haven't to throw garbage on the edge of the lake and liquid waste of potato farming banned to entered into the lake. That rules does not directly impact on the conservation of Cebong Lake. However, if these activities are strictly obeyed and implemented, there would be real impact on the existence of Cebong Lake. The most lately prominent issue in the Cebong Lake which initiated by the society and the government is making of integrated waste irrigation channels of potato farming. The irrigation channels are at the edges of the Cebong Lake and drain off the waste directly into the outlet of Cebong Lake so it could not enter the lake. The manufacture of irrigation channels are an integration between local wisdom and technology. Technology makes local wisdom more optimally to provide more real benefit. The benefits of irrigation channels are the sediment and pesticides does not enter into the lake so the condition of lake morphology still natural and far away from sedimentation which makes silting the lake. If the silting of the lake can be prevented, it can be said that Cebong Lake can be more sustainable. 4. Physical Characteristic Sembungan is the highest villages on the Dieng plateau. The village is located at an altitude of 2300 m above sea level and the climate is cold with temperatures 14-23°C. The most productive land for the cultivation of potatoes are on the slope 2-15 degrees which covers 15.77% region with topography wavy until hilly, which is around Cebong Lake (Arbangiyah, 2012). The first interpretation of physical characteristic using satellite image produce the area of Cebong Lake which is 84847,85 m2. In addition, bathymetry

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measurement produce the bathymetric map of Cebong Lake (Fig. 1). It show that average of the depth of Cebong Lake is 2,179 m. Results of laboratory tests on variables TSS and TDS (Table 1) shows that the water of the lake is still normal. Compared with water quality standards

of freshwater fish farming, livestock, and irrigation (PP 82 in 2001), the water is still under standard. These data indicate that the function of irrigation canals were built around the lake was optimal in order to prevent the entry of sediment into the lake.

Figure 1. Bathymetric map of Cebong Lake. Source: Field Measurement (2015) Table 1. Distribution of Suspended Sediment in Cebong Lake No. Depth Sample’s depth 1 2.5 0.8 0.2 2 3.5 0.8 0.2 3 1.75 0.8 0.2 4 2.6 0.8 0.2 5 1.5 0.6

TDS (mg/l) 196 244 212 172 128 232 192 212 216

TSS (mg/l) 30.1 18 20.2 21.3 12.3 9.2 14.8 34.3 24.8

Source: Laboratory Analysis (2015).

5. Conclusion As an eminent problem, sedimentation in lake can‟t be solved without local people contribution. This research shows that sediment rate in Cebong Lake is not apprehensive. Nowadays agriculture system in Cebong lake environment is comprehensive with drainage system which minimalized sediment enter the lake. Drainage system building by local people is one result of technology and local wisdom

integration. This integration may preserve Cebong Lake and impede its disappearance. Acknowledgements This study was financed by Student Creativity Program organized by Directorate General of Higher Education (DIKTI) Indonesia. In addition, authors will say big thanks for our professor, Prof. Dr. Sudarmadji M.Eng.Sc. The authors wish to acknowledge very useful and constructive comments by reviewers of this paper. References [1] Andriana, R. 2007. Evaluasi Kawasan Lindung Dataran Tinggi Dieng Kabupaten Wonosobo. Thesis. Environmental Science, University of Diponegoro, Indonesia. [2] Arbangiyah, R. 2012. Perubahan Pola Pertanian Rakyat di Desa Sembungan Dataran Tinggi Dieng (1985-1995), Mini Thesis, University of Indonesia.

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[3] Bruinsma, R. 1997. Hydrometry, Departemen Pekerjaan Umum, Jakarta, Indonesia. [4] Manalu, A. 2012. Kajian Sedimentasi Waduk Saguling, Provinsi Jawa Barat. Thesis. Magister of Natural Disaster Management, Gadjah Mada University, Indonesia. [5] Nursa‟ban, M. 2008. Evaluasi Sedimen Yield di Daerah Aliran Sungai Cisanggarung Bagian Hulu Dalam Memperkirakan Sisa Umur Waduk Darma. Jurnal Penelitian Saintek, Vol 13 No 1, April 2008: 47-64. [6] Pusat Litbang SDA. 2012. Pengelolaan Danau Dan Waduk Di Indonesia. Balai Lingkungan Keairan, Jakarta, Indonesia. [7] Santosa, I. 2011. Pengelolaan Sumberdaya Alam Dan Lingkungan Hidup Berbasis Kearifan Lokal, in proc. Seminar Nasional Hari Lingkungan Hidup 2011, LPPM University of Jenderal Soedirman, Indonesia. [8] Setiawan M. A.. 2012. Integrated Soil Erosion Risk Management In The Upper Serayu Watershed, Wonosobo District, Central Java Province, Indonesia, Dissertation, Institute of Geography, University of Innsburck. [9] Suyono, 2004. Hidrologi Dasar, Faculty of Geography, University of Gadjah Mada, Yogyakarta, Indonesia. [10] Thomas, R., Meybeck, M., Beim, A., 1992. Lakes: Water quality assessment- A guide to use biota, sediments and water environmental monitoring. UNESCO.

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Sanitation Condition and Potential Recovery of Nutrients in Urban Area of Sub District Kiaracondong, Bandung City, Indonesia Neni Sintawardani*, J. Tri Astuti*, Dewi Nilawati*, And Ken Ushijima** *Industrial and Environmental Physics Division, Research Center for Physics, Indonesian Institute of Sciences (LIPI), Jalan Cisitu-Sangkuriang, Bandung, 40135, Email: [email protected] **Tokyo Institute of Technology, Dep. Of Environmental Science and Technology, 4259, Nagatsuta-cho, Midori-ku, Yokohama 226-8502, Japan Abstract : A study was carried out to observe the sanitation conditions in the urban area of Kiaracondong and to estimate the potential for nutrient recovery from human excreta. Interviews about sanitation, covering the frequency of defecation, toilet facilities, water sources, and discharge systems, were conducted directly with 111 households through cooperation with the household association. Samples of fresh feces and urine from respondents were collected, weighed, and characterized. Data on the population of Kiaracondong sub-district was required to calculate the nutrient recovery potential based on the characteristics of the human excreta in the community studied. The data showed that all public toilets in Kiaracondong were installed along canals or rivers without water facilities and poor sanitation conditions. The wastewater from all public toilets and most private toilets were discharged directly to water bodies. The existence of public toilets is important and useful to the community in Kiaracondong. In total, the potential for the recovery of nutrients in human excreta were 57.32, 2.79, 1.57, and 1.65kg.p-1.y-1, for OM, N, P, and K respectively. For the Kiaracondong area, the potential was estimated to be 7,199, 350, 197 and 207 tons.y-1 for OM, N, P, and K respectively. There were no worm ova detected in the feces and urine samples. However, E. coli a pathogen and Enterobacter agglomerans and fungus of Candida sp were detected in the feces samples. Meanwhile, organisms detected in urine were E. coli a pathogen, Enterobacter aglomerans, Serratia liquefaciens, Serratia marcescens, and Staphylococcus saprophyticus. Implementing dry composting toilets based on the Ecosan concept is an appropriate way to address the sanitation problem. The education and promotion of Ecosan to increase the awareness of the community is important. Keywords: dry-toilet, human excreta, ecological sanitation, nutrient flow, recycle. 1. Introduction Population growth and rapid urbanization is a major pressure on urban regions. Rapidly growing cities face problems with the supply of infrastructure and public water and sanitation services. In some developing countries, wastewater treatment has not yet become affordable due to limited financial capabilities (Bömh et al., 2011). On the other hand, according to the Millennium Development Goals, the indicator for sanitation is focused on the availability of private latrines and safe disposal of human excreta (WHO, 2006). The amount and composition of human excreta that has been reported varies greatly from person to person. This depends on the diet and habits of the population (Josson, 1997). Treating human excreta is required to reduce the spread of communicable disease and prevent the pollution of surface and groundwater due to its potentially high concentration of pathogens (WHO, 2006). Wastewater containing excreta is a hazard to hygiene, and also contains organic matter and eutrophying substances in the form of plant macronutrients, i.e. nitrogen (N), phosphorus (P) and potassium (K). These substances that originate in the urine and feces cause problems in water bodies. On the other hand, these can be

recycled and reused as biofertilisers that are valuable for agricultural purposes (Esrey et al., 2001). Through this approach, a lot of energy could be conserved by replacing chemical fertilisers with substances in urine and feces (Esrey et al., 2001; Jonsson & Vinneras, 2004; Vinneras et al., 2006). Approximately 45% of the Indonesian population lives without easy access to private toilets that are safe for urinating and defecation. Sewer facilities are commonly limited (Anonymous, 2008). A previous study has reported that only 4% of the population in the Citarum catchment area (a major river in West Java) discharged their domestic water to a septic tank, 5% to a wastewater treatment plant, and 91% directly to a channel or river without treatment, causing a significant water degradation (Sudjana et al., 2006). Kiaracondong, an urban slum area, is one of the subdistricts of Bandung city in the West Java province of Indonesia. It has the highest population density, 36,941 persons per km2. The average population density of Bandung is 13,792 persons per km2. It was reported (Astuti et al., 2006) that > 40% of the employees in Kiaracondong worked in the informal sector, with jobs such as coolies and small traders. According to the Indonesia Central Bureau of

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Statistics, about 30% of the population in Kiaracondong lives under the poverty line. Due to the shortage of space, limited income of the community and lack of availability of sanitation infrastructure, most households in Kiaracondong discharge all their domestic wastewater, both grey and black, directly into the Jondol canal, a part of the Citarum river catchment area, without any treatment. This can be observed by the outlet pipes that have been constructed by most households for discharging domestic wastewater (black and grey). As a result, water quality and environmental sanitation is decreased. Visually, the water of Jondol canal is dark, dirty and has a bad odour. The Ecosan concept seems to be an attractive way of solving sanitation problems. The basic principle of Ecosan is to close the loop between sanitation, water resources, and agriculture to reduce health risks, prevent pollution of surface and groundwater, prevent the degradation of soil fertility, and to optimize nutrients and water resources management. Human excreta will be recognized not as a waste, but as a resource that should be made available for reuse (Werner et al., 2003; Langergraber & Muellegger, 2005). A key option to reduce the nutrient load would be via optimizing waste collection and investment in dry or low-flush toilets (Erni et al., 2010). The dry-composting toilet, which uses neither water nor sewage infrastructure, is a practical solution in areas with inadequate sewage disposal and limited water. This system is promoted to sanitize and recycle human excreta into fertilizer (Redlinger et al., 2001). Nevertheless, the methods of waste collection, treatment and reuse should be compatible with local habits and social-religious practice (Nawab et al., 2006). This study was carried out to observe the sanitation conditions and calculate the potential for recovering nutrients from human waste in the urban slum area of Kiaracondong related to the concept of Ecological Sanitation (EcoSan). 2. Materials and Methods 2.1 Date and Site This study was carried out from 2005 to March 2006 in RW2 of Sukapura village, Kiaracondong subdistrict, Bandung city, West Java province, Indonesia. Kiaracondong sub-district is about 20 km southeast of Bandung city. It covers six villages, one of which is Sukapura. Sukapura village comprises eight Community Associations (RW: Rukun Warga) with each RW normally covering six to eight Neighborhood Associations (RT: Rukun Tetangga). Each RT usually consists of 50 to 60 families. The map of Bandung city, sub-district Kiaracondong and the study area in RW2 of Sukapura village is displayed in Figure 1. This study was conducted in coordination with the local government, including

the RW and RT, and by mobilizing the housewife association (PKK: Pembinaan Kesejahteraan Keluarga), mainly in RW2.

Figure 1. Bandung city, sub-district Kiaracondong, and RW2 of Sukapura village 2.2 Sanitation facilities and human defecation Sanitation facilities for human defecation, including toilets, sources of water and blackwater discharge systems was directly observed at sampled households, combined with interview data. The interviews were carried out in cooperation with PKK. 111 families were mobilized, spread out in eight RTs. This represented 27% of the families of RW2 in Sukapura village. Interview respondents filled up questionnaires about their frequency of daily defecation and urination. This data was required to calculate the rate of production of feces and urine, expressed in grams per person per day (g.p-1.d-1). 2.3 Sampling and analysis of human excreta Plastic bags were prepared for both urine and feces samples, coded and weighed to determine the initial weight of the plastic bags before using it for the packing of samples. The bags were sent to respondents for the packing each of fresh urine or feces samples in the the next morning. This method was implemented based on a previous study which indicated that most respondents usually defecated in the morning. Then, in coordination with PKK, all the samples of fresh feces and urine from the respective respondents‟ houses were collected and brought to the field station to be weighed. For each sample, the following were recorded: name, age, gender, and

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address of the respondent, date of sampling and weight of the sample (urine and fees). All the fresh feces and urine samples were chemically and microbiologically analysed. Chemical parameters include pH, organic matter (OM), N and P. Microbiological parameters include total plate count (TPC), bacteria, fungus and worm ova. Analysis was done at the Health Laboratory, Health Department of Indonesia in Bandung. 2.4 Potential for nutrient recovery from human excreta Secondary data covering the population of Kiaracondong sub-district (Center Bureau of Statistic, 2005) was required in order to calculate the potential for the recovery of nutrients based on the characteristics of human excreta of the community in this area. This was calculated by combining a range of data, including the rate of production of human excreta, its nutrient composition, population and the percentage of domestic wastewater discharged directly to Jondol canal. The nutrient recovery potential is defined as the nutrients in human excreta discharged directly to the canal that can be recycled and reused for agriculture. 3. Results and Discussion 3.1 Sanitation facilities discharging system

and

black

water

The interview results indicated 95% of the respondents prefer to use the squat type of toilet, with 5% preferring a seat type of toilet. The frequency of defecation, time of defecation, sanitation facilities and discharge of blackwater is presented in Figure 2.

Figure 2. Defecation and wastewater discharge system With regard to the habits of the respondents, the frequency of defecation was once a day (84%), twice a day (2%) or irregularly (9%). Most respondents defecate in the morning (77%). The data shows that 28% of the respondents had no toilet facilities in their house and therefore use public toilets. The distance to the public toilet from their house varied from less than 10m (42%), 10-50m (52%) and > 50m (6%). 57% of the households discharged blackwater directly to the canals or river, 38% to a septic tank and 5% dumped in soil. It could be predicted that more than 57% pollution in water bodies was caused by excreta/faeces which discharged directly to the canal or river and because of the improper construction of septic tanks. Since limited availability of open space in the location, most of existing septic tanks were constructed without proper consideration of health regulations. The distance between most septic tanks to wells was less than 10 m. This situation caused pollution to surface water and groundwater. All public toilets in Kiaracondong those were installed along Jondol canal to Cidurian river (in the catchment area of Citarum river) had no septic tanks and no proper water facilities. Sanitation conditions are poor. The toilet bowl was made of cement with very simple construction. Wastewater from all public toilets and most household toilets were discharged directly to the canal or river as presented in Figure 3. A direct observation of public toilet usage over 24 hours showed a high usage of 113p.d1 in RW7 and 87p.d-1 RW2 . Even the peak of public toilet usage was in the morning, mainly by

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students and employees before school and work, but the pattern was different as from the interview‟s data. Public toilet‟s usage fluctuated between 4 am to 8 pm (Sintawardani et al., 2005). This high

frequency of public toilet usage and distribution of time of usage indicates that its availability is important and useful for the community in Kiaracondong.

Figure 3. Public toilet (A, B), cement squat bowl (C), and discharging to canal (D). 3.2 Production rate of feces and urine In this study, 15 samples of feces were collected from respondents, comprising 12 women (80%) and three men (20%). The ages of the respondents are the range of 24-57, an average of 44.8 years old. Based on the interview results, most respondents (84%) defecate once a day. Therefore, as presented in Table 1, the rate of production of fresh feces varies from 55-288g.p-1.d-1, with an average of 120.6g.p-1.d-1. This result was lower than a previous study that reports the product rate of wet feces in

developing countries as 350g.p-1.d-1 in rural areas and 250g.p-1.d-1 in urban areas (Feachem et al., 1983); and in China it was reported as 315g.p-1.d-1 (Gao et al., 2002), but in Thailand it was in the range of 120-400g.p-1.d-1 (Schouw et al., 2002). The disparity in the rate of product of human excreta is affected by the difference in amount of food intake and composition of the food consumed. Consumption of food low in fiber results to smaller amount of mass of feces than food high in fiber.

Table 1. Production rate of fresh feces and urine for women (W) and men (M) in study

The quantity of urine excreta depends on how much a person drinks and sweats, their diet, physical activity and the climate (Feachem et al., 1983). Excessive sweating results in concentrated urine while the consumption of large amount of liquid dilutes the urine. Based on assumption if the frequency of urination per person is eight times per day, it could be calculated that the urine production rate is in the range of 696-3,040g.p-1.d-1 with an average of 1,294g.p-1.d-1 (Table 1). This result was similar to previous reports that urine production for most adults was in the range of 1,000-1,300g.p-1.d-1 (Feachem et al., 1983). In Sweden, the average production of urine is reported as 1,500 (Vinneras et al., 2006) and in Thailand it is 600-1,200g.p-1.d-1 (Schouw et al., 2002). 3.3 Composition of human excreta Characteristics of human excreta in the study are presented in Table 2 for feces and Table 3 for urine. The data showed that the pH of fresh feces was in the range of 5.42-7.19 with an average of 6.71. The

average moisture content of fresh feces was 76.95% (71.5382.68%). Correspondingly, the dry matter (DM) content is 24.05%. This result was in line with an earlier study by Polprasert (1995) that measuring water content of feces to 70-85% and DM to 1530%. The OM content varied from 62.37-89.69% DM or 90.84% DM on average. The total N content of feces in this study was in the range of 2.8-5.90% or 3.83% on average. The P content achieved was between 2.47-5.12% with an average of 3.50% and for K it was 0.98-3.70% with an average 2.33%. Related to the nutrient content of human feces, there is a disparity due to the varying food habits of respondents. An earlier study reported that fresh human feces contained 5.0-7.0% of N, 3.05.4% of P and 1.02.5% of K of DM (Mara, 1976). Another study reported that the N content of feces was 3.6% and the P content is 0.9% DM (Polprasert, 1984). The difference in the N content of feces might be caused by the variation of food consumed among the respondents, both in quantity and quality. The N content of the feces in this study (3.83%)

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closed to the research that was conducted in Thailand (3.6%). Traditionally and based on superficial observations on-site, people in West

Kiaracondong consumed less protein and more vegetables in their daily meals.

Table 2. Composition of feces samples (N=9)

Table 3 shows that the pH of urine was in the range of 4.93-6.99 with the average to 5.77. The DM content of fresh urine is 5.03-9.84 or 7.77% on average. The OM content was 67.37-89.69% or 82.24% on average. The IOM content was 17.76%

on average. The result showed that total N content of urine was 3.11-5.13 (average 4.11%); total P was 1.47-2.61% (average 2.43%DM), and; total K was 2.38-2.44% or 2.41% on average.

Table 3. Composition of urine samples (N=13)

Table 4. Calculated estimates of nutrient recovery from human excreta

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Nutrient recovery potential Based on the average production rate of fresh feces of 120.6g.p-1.d-1 as presented in Table 1, the average DM content is 24.05%, and combined with the nutrient content as mentioned in Table 2, the potential production of OM; N; P; and K of feces in this study is 9.62; 0.41; 0.16; and 0.25kg.p-1.y-1, respectively. Then, by considering the production rate of urine in Table 1 (1,294g.p-1.d-1) with a DM content of 12.28%, and its nutrient composition as presented in Table 3, the potential production of OM; N; P; and K of urine in

study are 47.70; 2.38; 1.41; and 1.40kg.p-1.y-1 respectively. The potential recovery of N from urine in the study is lower than a previous study, which reported the N and P content of urine as 2.5-4.3kg and 0.4-1.0kg.p-1.y-1 respectively (Vinnerâs et al., 2006). Table 4 presents the estimates of nutrient recovery of some previous studies (Jonsson & Vinneras, 2003) in comparison to this study result. The gap of nutrient recovery might be caused by differences in habits, types and amounts of food consumed.

Table 5. Population of Kiaracondong sub district by villages in year 2005 (persons)

Table 6. Estimates of nutrient recovery from human excreta in Kiaracondong area

The data showed that population of Kiaracondong sub-district in the year 2005 was 125,600 persons (Table 5) and it could be assumed 57% of the population discharged toilet waste directly to the water bodies. By combining these data with the production rate of nutrients from human excreta, it is estimated that the OM; N; P; and K recovered would be 1,208; 51; 20; and 31tons.y-1 from feces; and 5,991; 299; 177; and 176tons.y-1 from urine, respectively. In total, the nutrients from feces and urine that could potentially be recovered in the Kiaracondong urban area are 7,199; 350; 197; and 207 for OM; N; P; and K respectively (Table 6). The 3.4 The existence of organisms in human excreta samples Table 7 shows that worm ova were not detected in the feces nor urine samples. Fungus of Candida sp was found in 80% of feces samples, but not in urine. As predicted, bacteria of E. coli a pathogen were detected in all samples, but only one person‟s urine content E-coli. Meanwhile, Enterobacter aglomerans was only detected in the feces of a person and in other one person‟s urine. A higher variety of microorganisms was detected in urine, but bacteria of E. coli a pathogen was only detected in 20% of urine samples. The same result was found for Enterobacter aglomerans, Serratia liquifaciens, and Serratia marcescens. In contrast, Staphylococcus saprophyticus existed in all of the urine samples. It has been reported that the

previous research asserted that the discharge of feces and urine to water bodies caused water degradation due to its organic matter content and plant nutrient content. On the other hand, these nutrients would be valuable for agricultural purposes (Esrey et al., 2001), especially, macronutrients N, P, K that can be utilized instead of artificial fertilizer (Vinneras et al., 2004) and would be possible to recycle nutrients from human urine by using a constructed food chain (Adamsson, 2000). The organic materials increase the humus content and the water holding capacity of soil (Esrey et al., 2001). pathogens and parasites found in human excreta are widely responsible for a variety of diseases in developing countries (Prüss et al., 2002). Table 7 The existence of organisms in feces and urine samples The existing organisms E. coli a pathogen Enterobacter agglomerans Candida sp Worm ova The existing organisms E. coli a pathogen Enterobacter agglomerans Serratia liquifaciens

Feces samples W1 W2 W8 W9 M10 + + + + + -

+

-

-

-

-

+

+

+

+

Urine samples W1 W2 W8 W9 M10 + -

-

+

-

-

-

-

-

+

-

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-

-

-

+

+

+

-

-

-

Conclusion The wastewater from all public toilets and most private toilets was discharged directly to canals and river. The existence of public toilets is very important and useful for the community in Kiaracondong. All public toilets in Kiaracondong are installed along canals or river with no water facilities and poor sanitation conditions. Using the interview‟s data and sampling and analysis of human excreta, the potential production of OM, N, P, and K are 9.62, 0.41, 0.16, and 0.25kg.p-1.y-1 from feces, and 47.70, 2.38, 1.41, and 1.40kg.p-1.y-1 from urine. Totally, nutrients in human excreta that could be recovered are 57.32, 2.79, 1.57 and 1.65kg.p-1.y-1 for OM, N, P and K respectively. Total potential for nutrient recovery from the Kiaracondong could be estimated to be 7,199, 350, 197 and 207tons.y-1 for OM, N, P and K respectively. There was no worm ova detected in feces and urine in study. But, bacteria of E. coli a pathogen and Enterobacter aglomerans and fungus of Candida sp were detected in the feces samples. Meanwhile, the organisms that exist in urine are bacteria of E. coli a pathogen, Enterobacter aglomerans, Serratia liquifaciens, Serratia marcescens, and Staphylococcus saprophyticus. Based on the study, the possibility to apply dry composting toilet technology in such slum areas is very recommendable. The implementation of Ecosan-based dry composting toilet is an appropriate way of solving the sanitation problem in Kiaracondong. Education and promotion of this technology to increase the awareness of community is important. Acknowledgement This study was funded by the Indonesian Institute of Sciences (LIPI) and supported by Japan Science and Technology Agency (JST) in accordance to the joint research on sustainable sanitation. Grateful thanks to the officials in Kiaracondong sub district, Sukapura village, RW2 community, and PKK for their good cooperation. Thanks to Ibu Mimin and Bapak Adenan for the valuable assistance. References [1] Adamsson, M. 2000. Potential use of human urine by greenhouse culturing of microalgae (Scenedesmus acuminatus), zooplankton (Daphnia magna) and tomatoes (Lycopersicon). Ecological Engineering 16 (2000) 243–254

[2] Anonymous. 2008. Economic Impacts of sanitation in Indonesia. Research Report August 2008. Water and Sanitation Program (WSP) East Asia & Pacific. World Bank Office, Jakarta. [3] Astuti, J.T., Nilawati, D., Sintawardani, N., 2006. Public Toilet Utilization in Kiaracondong: A Preliminary Study of BioToilet Application (In Indonesian). Jurnal Yeknik Lingkungan. Special Edition August 2006: 111-118. [4] Bömh, H.R., Schramm, S., Bieker, S., Zeig, C., Anh, T.H., Thanh, N.C. 2011. The semicentralized approach to integrated water suppl.y and treatment of solid waste and wastewater-a flexible infrastructure strategy for rapidly growing urban regions: the case of Hanoi/Vietnam. Clean Tech Environ Policy. 13: 617-623. DOI 10.1007/s10098-0110378-9. [5] Erni, M., Drechsel, P., Bade, H.P., Schedegger, R., Zurbruegg, C., Kipfer, R. 2010. Bad for environment, good for the farmer? Urban sanitation and nutrient flows. Irrig Drainage Syst 24: 113-125. DOI 10.1007/s1079500909083-9. [6] Esrey, S., Gough, J., Rapaport, D., Sawyer, R., Simpson-Hebert, M., Vargas, J., Windlad, U. (Eds). 1998. Ecological sanitation. Stockholm, Sweden: SIDA 586 76 12 0 [7] Feachem, R.G., Bradley, D.J., Garelick, H., Mara, D. 1983. Sanitation and Desease: Health aspects of excreta and wastewater management. World Bank Studies in Water Supply and Sanitation 3. Pitman Press, Bath, GB. [8] Gao, X., Shen, T., Zheng, Y., Sun, X., Huang, S., Ren, Q., Tian, Y., Luan, G. 2002. Practical manure handbook. Chinese Agricultural Publishing House, Beijing, China. [9] Jonsson, H., Vinneras, B. 2004. Adapting of nutrient content of urine and faeces in different countries using FAO and Swedish data. Peer reviewed paper in the proceedings of the 2nd International Symposium on ecological sanitation, incoorperating the 1st IWA specialist group conference on sustainable sanitation. Division 44, Environment and Infrastucture sector project ecosan. 7th-11th April 2003. Lübeck, Germany. GTZ. [10] Jonsson, H. 1997. Assesment of sanitation systems and reuse of urine. In: Drangert, J.O., Bew, J., Winblad, U. editors. Ecological alternatives in sanitation. Proceedings from SIDA Sanitation Workshop, 6-9 August 1997, Berlingham, Sweden, p: 11-12.

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[11] Langergraber, G., Muellegger, E., 2005. Ecological Sanitation-a way to solve global sanitation problems? Elsevier. Environment International. 31: 433-444.

[23] WHO-guideline for save reuse of wastewater, excreta and greywater. Vol. 1-4.

[12] Mara, D. 1982. Sewage Treatment in Hot Climate, ELBS edition, JWS. [13] Nawab, B., Nyborg, I.L.P., Esser, K.B., Jenssen, P.D. 2006. Cultural preference in designing Ecological sanitation systems in North West Frontier Province, Pakistan. Journal of Environmental Psychology. 26 (3): 236-246. [14] Polprasert, C. 1984. Human waste recycling under tropical condition. Waste Management & Research. 2: 5361. [15] Prüss, A., Kay, D., Fewtrell L., Bartram J. 2002. Estimating the burden for disease for water, sanitation, and hygiene at a global level. Environ Health Perspective. 110 (5): 537-42. [16] Schouw, N.L., Danteravanich, S., Mosbaek, H., Tjell, J.C. 2002. Composition of human excreta-a case study from Southern Thailand. The Science of the Total Environment. 286:155-166 [17] Sintawardani, N., Irie, M., Triastuti, J., Ishikawa, T., and Affendi, M. 2005. Feasibility of introducing the bio-toilet to decrease the water pollution in an open canal: A case study in Kiaracondong slum area in Bandung City. P.101-110. In X.C. [18] Wang and R. Cheng (Eds.). Future of Urban Wastewater Systems – Decentralisation and Reuse. Proceedings of International Water Assosiation (IWA) Conference, Xi‟an China, 18-20 May, 2005. [19] Sudjana, P. 2006. Quality Status of River Water in Indonesia (in Indonesian). Vol. I, ITB, Bandung, Indonesia. [20] Redlinger, T., Graham, J., Barud, V.C., Avitia, R. 2001. Survival of Fecal Coliforms in DryComposting Toilets. Appl Environ Microbiol. 67 (9): 4036–4040. American Society for Microbiology. DOI 10.1128/AEM.67.9.40364040.2001. [21] Vinneras, B., Palmquist, H., Balmer, P., Jonsson, H. 2006. The composition of household wastewater and biodegradable solid waste-proposal for new norms for the flow of nutrients and heavy metals. Urban Water J. 3: 3-11. [22] Werner, C., Otterpohl, R., Jonsson, H. 2003. 10 recommendations for action from the Luebeck symposium on ecological sanitation. GTZIWA. WHO. 2006. 1st International Conference on Appropriate Technology Development 2015, Bandung, October 5th-7th, 2015 | 176

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The Potency of Cashew Kernels Processing Activity to Encourage Community Venture in Southwest Sumba Regency in East Nusa Tenggara Province Febtri Wijayanti, Fithria Novianti, Carolina Centre for Appropriate Technology - Indonesian Institute of Sciences Jalan KS Tubun No. 5, Subang 41213. Indonesia E-mail : [email protected] Abstract: Indonesia is the 5th biggest cashew nut producer after India, Vietnam, West Africa, East Africa, and Brazil. The production reached 180.000 tons in 2013, 50.000 tons of it, valued almost US$70million, and was exported. One of the potential producers of cashew nut is the district of Southwest Sumba. Initially planted in farmers land as reforestration tree in since early 1980s, it covers 14.688 hectares of area and majority of it are in Kodi, North Kodi, KodiBalaghar, and Tambolaka. Despite the simple technology implementation, cashew nut becomes one of family income source through creation of many small businesses in South West Sumba. The technology utilized limits to shelled cashew handling, stripping, and frying. In proportion, cashew nut sales are 90% in shelled and 10% are processed. Supply chain analysis indicates that income derived from the businesses in the range of US$3,360 for the sale of cashew with shelled, and US$4,036 per year for the sale of processed cashew. The IRR’s reach 12,89% and the NPV are IDR 20.754.611 with the assumption of 9% discount factors (KUR Rate). From business point of view, the investation is feasible, however it has no significant contribution to economic growth in the community. The picture revealsthat small business based on cashew nut is not a sufficient to provide prospective income generating for society in East West Sumba. It indicates the need of government intervention to establish government owned business based on cashew nut production could be the solution to reach equity of community welfare. Keywords: appropriate technology, community prosperity, micro business, South West Sumba. 1. Introduction Indonesia is one of the most important cashew producers in the world, asIndia, Vietnam, West Africa, East Africa and Brazil are.Cashew is a drought resistant crop that can grow successfully in areas with a very distinct dry season or where the annual rainfall is as low as 50 cm. It can also grow well in areas with high levels of rainfall (as much as 350 cm annually) provided the soil is well drained. Cashew grows >12 m high and has a spread of 25 m. Its extensive root system allows it to tolerate a wide range of moisture levels and soil types (Olife, Jolaoso, Onwualu, 2013). In Indonesia, cashew most planted in eastern region. Approximately 554,315 hectares of land is cultivated with cashew, and 553.166 hectares of 99,7% of it are smallholders plantation producing 99,8% of the total production of Indonesia or 115,969 tonnes of dried cashew nuts(Ministry of Agriculture, 2015). As much as 52,263 tonnes cashew worth US $ 90,795,000 were exported in 2013, only 3,755 tonnes were imported. Of those, East Nusa Tenggara contribute 180,567 hectares of land producing 39,360 tonnes of dried cashew, reaching the export value of US $ 90,795. At a glance we could see how powerful the role of cashew for its plantation owner. Referring to the fact that more than 90% cashew is contributed by smallholder,we conducted this study. Focussing on the case we found in Southwest Sumba Regency that reflects general situation of East Nusa Tenggara

Province. In the area where cashew is considered as one of the most important agricultural commodity for locals family earnings, the area still suffer from poverty (Statistics Office of Southwest Sumba, 2014). The fact is quite contradictory as cashew has attained the status of export oriented commodity for Indonesia, and Southwest Sumba Regency is one of the important contributor. Therefore, it is important to investigate possible chances to provide added value to cashew which at present being sold mainly in form of dried cashew nuts in shell. The idea of integrating appropriate processing technology to the product is ofcourse not an easy task to conduct, nonetheless valuable to explore further. 2. Methodology The data and information used in this study is collected through indepth interview to those who are in the supply chain management of cashew in Southwest Sumba area namely smallholders, processors, shop owner and extension service officer. We also did a field observation to gain more understanding of the phenomenon. This study conducted during period of May to July 2015. The data is then analysed to attain an accurate picture of the proposed business based on cashew nut processing performed by micro scale entrepeneurs. The information include: capital/ investment cost, production costs, fixed costs,

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variable costs, semi-variable costs, as well asother related data. Stages in analyzing the financial feasibility of production enterprises processing cashew nuts are : 1. 2. 3. 4. 5. 6.

Investment costs Production costs Financial Structure Sales Estimation Cashflow The fulfillment of financial eligibility criteria: Revenue- Cost Ratio (R-C ratio), Net Present Value NPV, and Incremental Rate of Return (IRR.

The data is processed with tabulation and analysis based on financial feasibility measure such as Revenue - Cost Ratio (R-C Ratio), Net Present Value (NPV), Incremental and Rate of Return (IRR). Variable costs and fixed costs are indicators used to determine the total cost of production or total cost, by calculation 𝑇𝑜𝑡𝑎𝑙𝐶𝑜𝑠𝑡 = 𝑉𝑎𝑟𝑖𝑎𝑏𝑙𝑒𝐶𝑜𝑠𝑡 + 𝐹𝑖𝑥𝑒𝑑𝐶𝑜𝑠𝑡 Determination of the assumptions made to support data analysis. Cost calculation was conducted on the investment cost, variable-semi-variable costs, fixed costs, and other expenses. Cost is the amount of capital investment or costs used to start a business orexpand a business. Variable costs are costs occuring in each routine production in which the amount depends on the amount of product you want to be produced. Fixed costs are other types of costs which are routinely issued by companies for corporate conduct production activities, but the amounts of fixed costs do not depend on the production capacity. BEP (break-even point) is total production or sale of the same, where it must be done so that the costs can be covered again or where the profit received by micro entrepeneurs is zero (Ibrahim, 2009).

𝐵𝐸𝑃 =

𝑇𝑜𝑡𝑎𝑙 𝐹𝑖𝑥𝑒𝑑 𝐶𝑜𝑠𝑡 𝑃𝑟𝑖𝑐𝑒 𝑜𝑓 𝑃𝑟𝑜𝑑𝑢𝑐𝑡 − 𝑉𝑎𝑟𝑖𝑎𝑏𝑒𝑙 𝐶𝑜𝑠𝑡

Analysis of Net Present Value is to estimate the value of investments by considering changes in currency value. NPV is the net present value which is the sum of all the future cash flows to determine the present value. Cash flows include the both inflows and outflows that are discounted at a rate (Arshad, 2012) NPV > 0, bussines is feasible NPV = 0, The business to restore as much value for money invested NPV < 0, business is not feasible to run

IRR is the “Internal Rate of Return” which is used to determine that what rate of return an investor is taking on a project. IRR describes what is the percentage return on investment. A business is considered feasible if the IRR is greater than Marginal Average Revenue Return (MARR). Whereas MARR can becalculated as the following equation (Kusumanto, 2008): 𝑀𝐴𝑅𝑅 = 1 + 𝑖 1 + 𝑓 − 1 In which: i = the interest rate investments f = the highest inflation 3. Cashew in South West Sumba : An overview Cashew (Annacardium officinalle) tree for people in East Nusa Tenggara is known since early 1980‟s as regreening plant species. The characteristics of cashew tree that can grow well in drought condition makes this plant is widely cultivated as reforestration tree in dry regions such as South West Sumba. Until presently, cashew trees grow extensively in Sub Districts area of Kodi, North Kodi, Kodi Bangedo, Kodi Balaghar and Laora. Covering 14,283 hectares of land, South West Sumba produced 4563 tonnes in 2013. It is a significant declination of productivity since in 2008, the area produced 6,055 tons of raw cashew nut in shell (South West Sumba Statistics, 2014). This production slides seems to relate strongly with the lack of proper treatment in cashew tree planting ecosystem. The ease to harvest cashew nuts over the years has pampered farmers to develop a false believe that the trees do not require thorough ecosystem management. Despite this situation, cashew stays as important cash crop for many South West Sumba farmers. Cashew trading system in South West Sumba is now still dominated by dry shelled nutsbuyers, most of them came from India. They buy raw cashew at a price higher than the price elsewhere. The buyers from India purchased the raw cashew at an estimated price of IDR 16,000 - IDR 22,000 per kg. Meanwhile, the price of raw cashew kernels, range from IDR 100,000 to IDR 105,000 per kg, whereas the price of fried cashew kernels is IDR 140,000 to IDR 180,000,- per kg. Buyers from abroad, such as India only interested to buy raw cashew not processed nuts because they need the whole shelled nut to be used as raw materials for many industries that produced various products. Southwest Sumba has been part of the value chain of worlds cashew trade, and this situation is actually detrimental as the added value is taken abroad. The price fluctuates freely depends on the quantity of cashew available. Consequently, sale price dropped significantly during harvest time, and this limits them from gaining good income. This is the result of leaving the farmers to become part of free

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trade dynamics in which price is dependant on the market. Without a good bargaining power, farmers sale is easily controlled by middleman. The economic pressure force farmers to go with whatever situation offered by the middlemen. Referring to this situation, we promotes the idea of providing added value to cashew nuts by introducing processing technology. Thus, farmers will have both chances to gain income, by selling raw shelled nuts and processed nuts. Innovation technology in cashew processing need to be introduction to farmers.Innovation in cashew handling will give back value added to local farmers. This study will valuate financial feasibility of processing of cashew nut. We hope, the analysis can be used by the local government to create and implement appropriate strategy to increase the potency of cashew as economic commodity to support family welfare. The most important thing to develop innovation of microentreprises are support. Support for innovation processes in SMEs requires a complex institutional framework in which the essential role is played by regional and local institutions (Lisowska & Stanislawski, 2015) Southwest Sumba, as one of important cashew producer in Indonesia, should be able to develop to increase community welfare. As a region who has a

poorest grade for poverty, cashew could be a strategy to community empowerment. We can take Thailand with their OTOP (One Tambun One Product) as an example of rural development strategy. OTOP Thailand is a government program that supports micro and small enterprises (MSEs), rural SMEs (small and medium-sized enterprises), cottage industry, local enterprises are characteristic of OTOP enterprise business for increasingly seen as the creators of new jobs. The OTOP policy has simplified the successful development of many MSEs. OTOP initiatives have enabled communities to combat poverty by value adding to natural resource products through integrating local knowledge and modern processing technologies (Phonsuwan& Kachitvichyanukul, 2010). 4. Financial Analysis of Cashew Potency 1.1 Initial Investment The investment costs are fixed costs which are not influenced by the amount of product. In cashew processing, the cost of the investment required, among others: kacip (cashew nut sheller), fryer, electrical sealer, scales, production house and bed dryer. Out of the cost and value in cashew processing, it is calculated that the investment cost is IDR 29,970,000. The detail analysis of the investment costs can be seen in Table 1as follows.

Table 1. Initial Investment of cashew Nut Processing Business Production Utilities Production Machine Cashew Nut Sheller Fryer Trays Plastic Sealer Steelyards 100 kgs Digital Scale 1 kg Spinner Oven Production House and the utilities Drying Bed 20x20m2 Production House SUM OF INITIAL INVESTMENT

Amount 2 2 6 1 1 1 1 1

unit package unit package unit unit unit unit

1 1

Package Package

Price per Investation Cost package (IDR) (IDR) 300.000 600.000 1.000.000 2.000.000 20.000 120.000 1.500.000 1.500.000 1.600.000 1.600.000 150.000 150.000 3.500.000 3.500.000 3.500.000 3.500.000

5.000.000 12.000.000

5.000.000 12.000.000 29.970.000

Lifetime (month) 24 24 12 60 60 24 60 60

120 120

Depreciatio n (IDR) 25.000 83.333 10.000 25.000 26.667 6.250 58.333 58.333

41.667 100.000 434.583

1.2 Operating costs Operating costs is the cost determined by the amount of product produced. Operational costs consist of fixed costs, variable andsemi-variable costs. Processing of cashew nuts require operationalcosts to include: raw material costs, labor costs,

administrative costs and promotion, the cost of electricity and water, transportation and communication, by also taking depreciation and maintenance costs into account. These cost can be seen in Table 2.

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Table 2. Cashew Seed Processing Operating Costs

V A R I A B E L

COST AMOUNT PRICE (IDR) COST OF RAW MATERIALS cashew with Shell 48,00 kgs/day 16.500 Packaging materials 1 package/day 35.520 Cooking Oils 0,6 litre/day 15.000 Fuels 2 bundle 5.000 Amount LABOR COST Permanent Labor 2 Person a day 30.000 Manager 1 Person a day 50.000 Cashew Sheller Labor 48,00 kgs/process 5.000 Amount

COST PER OPERATIONAL MONTH COST (IDR) 4batch/month (IDR)

792.000 35.520 9.000 10.000 846.520 60.000 50.000 240.000 350.000

COST PER MONTH 20 batch/month (IDR)

COST PER MONTH 25 batch/month (IDR)

3.168.000 142.080 36.000 40.000 3.386.080 240.000 200.000 960.000 1.400.000

15.840.000 710.400 180.000 200.000 16.930.400

19.800.000 888.000 225.000 250.000 21.163.000

1.200.000 1.000.000 4.800.000 7.000.000

1.500.000 1.250.000 6.000.000 8.750.000

ADMINISTRATION & PROMOTION COST

ls

50.000

50.000

50.000

ELECTRICITY & WATER COST

ls

200.000

200.000

200.000

TRANSPORTATION & COMMUNICATION COST FIXED DEPRECIATION COST COST MAINTENANCE COST SUM OF OPERATIONAL COST

ls ls ls

400.000 434.583 62.438 5.933.101

C O S T

1.3 The Need of Investment and Working Capital Funds Funds required to produce processed cashew kernels in exception of machinery and other equipment is called working capital. Based on our calculation, they will require approximately IDR 8,340,000 per month. Furthermore, the initial investment to start the business is IDR 29,970,000. 1.4 Production and Income Based on the data and information collected from micro-entrepeneurs who work with cashew in Laora Sub Districts - Southwest Sumba, the estimated amount of production and the average income of the cashew proccessing in micro entreprises are as shown in Table 4. The assumptions is constructed based on the adjusted price to the real price in 2015 in which the selling price of fried cashew kernels is IDR 180,000 per kgs.

400.000 400.000 434.583 434.583 62.438 62.438 25.077.421 29.913.000

1.5 Cashew Processing Business Analysis Based on an feasibility analysis of processing cashew, it is found that the IRR or Internal Rate of Return produced is 12,89% assuming a discount factor of 9% (KUR rate). However, the NPV for 5 years period of this business is relatively smaller than initial investment. This indicates that cashew processing business in the scale of household enterprise runs slowly. The value of IRR and NPV that are smaller than the value of this investment indicates that the business cannot provide a significant benefit to trigger the welfare of cashew farmers. Consideration of many significant supporting factors to develop a good business climate based on cashew kernels processing technology is utmost important. It seems that we can not leave the people alone struggling for the sustenance of their livelihoods. This is the area of which government intervention is required.

Table 4. Cashew Processing Sales Revenue

Selling Price Percentage of Weight (gr) (IDR) Sale Pack A 250 45.000 60% Pack B 500 90.000 20% Pack C 1000 180.000 20% Amount of Revenue (IDR/month)

Sales revenue (IDR/month) 25.920.000 8.640.000 8.640.000 43.200.000

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Table 5. Cash Flow Cashew Nut Processing Years 1 2 3 4 5 IRR NPV RC Ratio Payback Period (month)

Revenue (IDR) Expenses (IDR) 51.840.000 49.743.245 615.600.000 522.740.050 648.000.000 549.528.250 648.000.000 549.528.250 648.000.000 549.528.250 12,89% 20.754.611 1,16 12,00

The above figure indicates that that the cash balance of the micro-enterpreuneur stay stagnant from month to month. This condition illustrates that the cashew nut processing business is still traditionally operated. In reference to the case of Southwest Sumba, the business does not show any significant potency to increase the income of cashew processing microentrepreneurs. From the economic standpoint, this business can be regarded as subsistence, or "just sufficient to survive". It is not a highly profitable business However, apart from the economic

benefits there is also a social benefit to consider. In terms of employment they arenot considered unemployed, because they also obtained profits derived from labor wages providing opportunity for their family members to gain income. Papazov (2014) said that Small businesses typically have limited ability to mobilize own or attracted funds and, profits are usually sufficient only to support the owners‟ living and can contribute little to the selffinancingof the small company‟s activities.

Picture 1. Financial Status of Cashew Processing on Micro enterprises This situation is “usual” for startup microentreprises based on agriculture commodity, which is, Social cultural as a society influence in their business. Informal businesses are marked by seasonality not only due to the characteristics of the businesses, but also due to circumstances of households running them (Kamath & Ramanathan, 2013). In Southwest Sumba, there are many traditional ceremony, whichthe ceremony spending many household and entreprise asset. In that period, they lost their productive time and economic assets. As we know, micro entrepeneurs spesifically in rural, usually is not efficient company. They use bigger input and lower output.

1.6 Business Feasibility of Raw and Processed Cashew Nuts The fact that cashew kernels processing is not a popular technology to be utilized by micro entrepreneurs draw currently, cashew processing business needs to be more developed. The development is not merely for the purpose of conservation land, but to shift the economic interests of society at large. Transformation on the status of cashew, such as reforestation, tree planting species become the primary commodity, require a pattern of developmentof agribusiness. Therefore, the purpose of the development of agribussines of cashew is now directed to a) increase production and quality, both

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to meet domestic demand and export; b) increase the income of farmers and the expansion of employment opportunities; andc) maintaining and preserving the environment through reforestation programs and the rehabilitation of land degradation (Indrawanto dkk., 2003; Indrawanto, 2008; Listyati and Sudjarmoko, 2011). In Southwest Sumba, cashew nut processing is not a too popular knowledge of the people. They take the preference of selling dried cashew nuts in shell directly to in the buyer. Introduction of de-shelling technology and cashew nuts processing will stipulate a new mindset of attaining added value of their harvested cashew.Table 7 is shown the revenue comparison between selling dried cashew and processing cashew like fried cashew nut. Table 7. Revenue/Profit Comparison

Selling Fried Cashew Nut Selling Dried Cashew

Revenue (IDR/year) 54,486,152 45,360,000

Sumba. Linkage between cashew community is a answer of their economic problem. 5.2 Recommendation In order to encourage further to become community welfare enhancer, the micro scale enterprise of cashew nut processing requires supports in many business aspects i.e. technology, management, and working capital in order to fulfill a much larger market share and more variety of market niche. Utmost important is creating and implementing strategic policy to manage the businesses in a form of community institution in which equity and equitability can be achieved. The families that operate microenterprises typically lack assets, especially marketable assets. Family members who operate microenterprises, especially women, have serious time constraints because of household responsibilities. These characteristics require substitutes for formal collateral, rapid loanprocessing procedures, and convenient locations for financial offices (Rhyne & Otero, 1994) Acknowledgment

For poor farmers, microentreprises are a source of income and employment where in their condition, there are no available alternatives. One possibility to improve the opportunities for the poor is to provide a favorable environment for Small and Medium Enterprises (SMEs). For example, micro, small, and medium enterprises (MSMEs) typically have high labor intensity and thus support job creation, especially in the industry and services sector (Rocha, 2012). Based on this study of cashew nut processing, to produce cashew snack, need much investment cost, especially for poor farmers, In addition to technology. Lacking of financial access, microentreprise often failed to growth. Unfortunately, these independent business groups, especially small and micro businesses that have low income are not easy to get capital or financing from the banking institutions due to lack of financial history. One of the funding solutions to small and micro businesses is microfinancing (Ibrahim & verliyantina, 2012).

The authors would like to convey gratitude to the Center of Appropriate Technology Development – Indonesian Insititute of Sciences that support the conduct of IPTEKDA Project in Southwest Sumba in 2015; and the Government of Southwest Sumba District who have provided assistances in support of this work. We also acknowledge the constant supports bestowed by Hari Siswoyo, Arie Sudaryanto, Bambang Saksana, Dominggus Bula, Marni Mada, Anderias Bulu for strengthening our understanding of the phenomenon.

5. Conclusionand Recommendation

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5.1 Conclusion Cashew processing in Southwest Sumba Regency is not a popular economic activity among locals. The underlying limitations on its adoption is the abiliy to penetrate market. Based on financial analysis of cashew nuts processing based business, micro scale seems feasible to become a family income generating activity, although not sufficient to promote community welfare in general. It need a huge effort and government support to promote community welfare for cashew farmers in Southwest

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