Available online at www.sciencedirect.com ScienceDirect Energy Procedia 65 (2015 ) 362 367 Conference and Exhibition Indonesia - New, Renewable Energy and Energy Conservation (The 3 rd Indo-EBTKE ConEx 2014) Evaluation of Protein Concentrate from Jatropha Seed Cake as a Soybean Meal Substitution in The Rabbit Feed Titin Widiyastuti a*, Tri Rahardjo Sutardi a, Roy Hendroko Setyobudi b a Animal Science Faculty of Jenderal Soedirman University, Purwokerto, 53121, Central Java, Indonesia b PT. Sinarmas Agroresources and Technology, Jakarta, Indonesia Abstract This research aimed is to evaluated post fermented protein concentrate from jatropha seed cake by using it as feed and the effect on blood metabolite of post weaned Rex rabbits (Oryctolagus cuniculus). Variables investigated were feed consumption and blood metabolite profiles (triglycerides and glucose). Analysis of variance showed significant effect on feed consumption and level blood glucose (P < 0.05), but not on blood triglyceride of post weaned Rex rabbit (P > 0,05). Based on feed intake and blood metabolite profiles, the experiment resulted that soybean meal can be substituted with protein concentrate from jatropha seed cake as a feed of post weaned Rex rabbits. 2015 Published T. Widiyastuti, by Elsevier T.R. Ltd. Sutardi, This is R. an Hendroko open access Setyobudi. article under Published the CC BY-NC-ND by Elsevier license Ltd. (http://creativecommons.org/licenses/by-nc-nd/4.0/). Peer-review under responsibility of the Scientific Committee of EBTKE ConEx 2014. Peer-review under responsibility of the Scientific Committee of EBTKE ConEx 2014 Keywords: Biorefinery; fermented protein concentrate; Jatropha curcas Linn. seed cake; rabbit (Oryctolagus cuniculus). * Corresponding author. Tel.: +62-816 4288 555; fax: +62-281-638 792. E-mail address: dyast72@yahoo.com 1876-6102 2015 Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). Peer-review under responsibility of the Scientific Committee of EBTKE ConEx 2014 doi:10.1016/j.egypro.2015.01.069
Titin Widiyastuti et al. / Energy Procedia 65 ( 2015 ) 362 367 363 Nomenclature BW Body Weight d day CP JSC Concentrate Protein of Jatrophha Seed Cake min minute CP-JSCF Concentrate Protein of Jatrophha Seed Cake Fermented DE Digestible Energy ME Metabolizable Energy RH Relative Humidity SBM Soy Bean Meal 1. Introduction Jatropha seed cake by products was obtained from oil refining jatropha seed, it has nutrients content equivalent to soybean seed cake. However, it is need an appropriate technology to optimize the utilization of seed oilcake due to its antinutrition factors. Jatropha seed cake has been yet using as animal feed whereas its potential in term nutrient composition. Chemical composition of toxically jatropha seed (dry matter basis) contain 56.4 % whereas non-toxic varieties 63.8 % crude protein [1]. Jatropha seed cake potentially using as feed, its related to nutrient composition and amino acid content. But there are obstacles in its utilization, reported that jatropha seed cake contain antinutritive factor viz. phorbol esters, trypsin inhibitors, lectins and phytate. Its restricted the use of jatropha seed cake as feedstuff. Concentrate protein from jatropha seed cake (CP-JSC) was an alternative feed stuff that has high nutrien potency for substitute the soy bean meal (SBM). The nutrient comparation between fermented CP-JSC and SBM showed in Table 1. Physic nut (Jatropha curcas L.) protein isolate was successfully achieved from jatropha seed cake by an alkaline extraction and followed by an isoelectric precipitation. The protein isolate had small amounts of phorbol esters, phytic acid, and saponin without any lectin. Its minimum and maximum solubility were at ph 4.0 and ph 12.0, respectively. Its water and oil binding capacities were 3.22 g water g 1 protein and 1.86 ml oil g 1 protein, respectively. Its foaming capacity and emulsion activity showed high values in a range of basic ph. Its foaming and emulsion stability values decreased with increasing time and exhibited high levels under basic ph conditions. Jatropha protein isolate had unique functional properties in water binding capacity, emulsion activity, and emulsion stability indicating an important role in food systems. It may be applied to salad dressing, mayonnaise, sausage, and meat products. Therefore, jatropha seed cake has a potential to be exploited as a novel source of functional protein for food or feed applications [2]. Reference [3] reported that digestibility and metabolizable energy of heat treated (121 C, 66 % RH, 30 min) kernel meal using the in vitro gas method were lower compared to those for soya bean meal (78 to 88) % and (11 to 13) MJ, respectively. Digestibility of jatropha kernel meal protein determined by treatment with pepsin, followed by tripsin was similar to that toasted soya bean meal (90 %). High nutrient potency of JSC could be enhance through, precipitation methods and continued by fermentation. This methods could reduce the antinutrient content i.e. phorbolester. Reference [4] reported that decreasing phorbolester and antitrypsin post fermentation of concentrate protein from Jatropha seed cake using Lactobacillus acidophilus. Munarso [5] reported that the content of phorbolester in the jatropha seed cake can be reduced by 32 % to 38 % after boiled or fermented with A.oryzae. Meanwhile Widiyastuti et al. [4] reported a higher rate of decline phorbolester on jatropha seed cake after precipitation and fermentation using Lactobacillus acidophilus is about 40.80 % (from 0.634 mg g 1 to 0.26 mg g 1 ). Table 1. Nutrient comparation between fermented CP-JSC and Soy Bean Meal Nutrien CP-JSCF a SBM b Moisture (%) 79.25 11.9 Crude protein (% ) 44.42 46.9 Crude fat (%) 12.56 2.66 Crude fiber (%) 1.31 5.90 ME (kcal kg -1 ) 2 814 2 230 Reference : a [4] b [6]
364 Titin Widiyastuti et al. / Energy Procedia 65 ( 2015 ) 362 367 Optimizing concentrate protein form jatropha seed cake as feedstuff could be done through optimation the protein potency and eliminate toxin content. This research aimed was : (i) to evaluated post fermented protein concentrate from jatropha seed cake by using it as rabbit feed and (ii) to analyze the level of fermented protein concentrate from jatropha seed cake and the effect on blood metabolite of post weaned Rex rabbits. Through this research, the use of concentrate protein was studied as rabbit feed for enhance jatropha cultivation value in the biofuel industry in Indonesia. 2. Material and methods 2.1. Material preparation and feeding trial Jatropha seed cake was got from PT SMART, Jakarta. Precipitation methods for produce concentrate protein according to reference [1] and [4]. Concentrate protein was fermented by L.acidophilus at 37 C for 3 d, then using as ingredient in rabbit feed (iso nitrogen and isoenergy), feed consist of four level of concentrate protein of Jatropha seed cake (CPJSC-F) were 0 % (R1), 6 % (R2), 12 % (R3) and 18 % (R4) or soy bean meal substitution was 0 %, 33.33 %, 66.67 % and 100 %. Twenty post weaned Rex rabbit - Oryctolagus cuniculus (the range of body weight were 600 to 1 000 g) was used and keep for 30 d. Pellet complete feed contain fermented concentrate protein was given twice a day (60 % in the morning and 40 % in the afternoon). Drinking water given ad libitum. 2.2. Feed ingredients and diet formulation The experimental diets were formulated to be iso-nitrogenous and isocalorific value. The nutrient requirement was according to reference [7] for growing rabbit (post weaned rabbit). Ingredient and composition by chemical analysis of the dietary treatment are shown in Table 2. Table 2. Nutrient composition of the experimental diets Nutrient composition (As feed basis (%) R1 R2 R3 R4 (%) Dry matter 88.83 88.97 89.12 89.26 Crude protein 17.36 17.37 17.15 16.52 Crude fat 6.35 6.84 7.34 7.84 Crude fiber 16.11 15.84 15.56 15.29 DE(kcal kg 1 ) 2 724 2 654 2 701 2 667 Ca 0.26 0.26 0.26 0.26 P 0.53 0.51 0.50 0.49 2.3. Analysis of blood metabolite profiles Blood sample was taken from cardiac with a spuit that containing Na-EDTA (Natrium Ethylen Diamin Tetra Acetic Acid) as an anticoagulant. Rabbit was fasted before blood collecting. Analysis of blood metabolite profile was according to reference [8]. 2.4. Statistic analysis Analysis of variance for feed consumption, blood glucose and triglyceride, using Adjusted SPSS 16 th version for tests. 3. Result and discussion The results showed the range of feed consumption (dry matter (DM) base) was 58.33 g (R1), 48.97 g (R2), 39.19 g (R3), 42.08 g (R4). The range of blood glucose was 58 g L 1 (R1) to 88.4 g L 1 (R3). The range of blood triglyceride was 99.998 g L 1 (R3) to 162.86 g L 1 (R2). Analysis of variance showed significant effect on feed consumption and level blood glucose (P < 0.05), but not on blood triglyceride of post weaned Rex rabbit (P > 0.05).
Titin Widiyastuti et al. / Energy Procedia 65 ( 2015 ) 362 367 365 Table 3. Feed consumption and blood metabolite profiles of rabbit fed concentrate protein of jatropha seed cake Level fermented CP JSC Dry matter intake *) Triglyseride *) Glucose *) R1 (0 %) (62.78 ± 20.73) g ( 101.66 ± 42.7) g L 1 (58.00 ± 11.47) g L 1 R2 (6 %) (41.49 ± 5.28) g (162.86 ± 44.89) g L 1 (81.60 ± 19.52) g L 1 R3 (12 %) (42.16 ± 5.12) g (100.00 ± 10.10) g L 1 (88.40 ± 4.66) g L 1 R4 (18 %) (43.96 ± 9.89) g (120.00 ± 70.43) g L 1 (69.00 ± 10.34) g L 1 Noted : *) (value ± se) g and (value ± se) g L 1 Feed consumption showed significant, with linear respons (y = -1.221x + 55.66; R 2 : 33.33 %). Based on the result, feed consumption was in the normal range (according to the body weight in this research). Ideal consumption (in dry matter basis) for rabbit is 65 g kg 1 to 80 g kg 1 body weight (BW) [9]. Low feed consumption is not only influenced by the feed ingredients, but also influenced by other factors. Based on the number of determination (R 2 = 33.33 %), it can be stated that the effect of CP-JSC fermented on feed consumption amounted to 33.33 %, the rest is influenced by other factors. According reference [10,11] the palatability of feed in cattle is influenced by several factors, both internal factors (habits, age and taste). and external factors (the nature of the feed and environmental conditions). Further explained that the palatability associated with smell, taste, and texture that can affect appetite. Fig. 1. Feed intake of rabbits fed containing fermented CP-JSC Serum Glucose concentration does indicate the gluconeogenetic capacity and or the degree of interference with ileal glucose absorption while serum triglyceride can be used as indicative of the degree of disruption of both ileal lipid absorptive and or homeostatic endogenous synthesis and or mobilisation of the metabolites [12]. Blood glucose of R3 (feed contain 12 % fermented CP-JSC) was the highest (88 g L 1 ), polynomial orthogonal assessment showed linear regression with equation y = 0.104x + 66.56 (R 2 : 3.9 %). Based on the results, it can be stated that the influence of the protein concentrate of JSC amounted to only 3.9 % on the blood glucose levels. The glucose levels in this research is lower than reference [13], It stated that blood glucose levels of local male rabbits weighing 1.3 kg to 2 kg were 137 g L 1 ± 10.15 g L 1. It was suggested that phorbolester content in the fermented CP-JSC lead to inhibit some enzymes activities. Reference [14] reported that the interaction of phorbol ester with protein kinase C (PKC) affects activities of several enzymes. Chivandi et al. [12] also reported that the decrease in Alpha Amylase with an increase in jatropha curcas meal in the diets could have been due to the suppression of the exocrine pancreas.
366 Titin Widiyastuti et al. / Energy Procedia 65 ( 2015 ) 362 367 Serum Triglycerides (STG) mainly come from dietary input through absorption, with some originating from adipose tissue catabolism. The decline in STG point was to poor to supply from gastro intestinal (GIT). Interplay of malabsorption or resulted from intestinal lesions characterized by decreased ileal absorptive surface area as a result of death of ileal mucosal enterocytes and or interference with venous or lymphatic drainage and or distension of the lamina propria cells [12]. Triglyceride level its suggested that there is no effects from fermented CP-JSC antinutrient of post weaned rabbit. Otherwise the highest triglyceride was showed by R2 (feed contain 6 % fermented CP-JSC) about 162.88 g L 1 ± 44.89 g L 1. The range of triglycerides level in this research was higher than reference [15] that showed range of triglycerides level of adult rabbit was about 47 g L 1 to 60 g L 1. It was suggested that high content of fat or digestible energy (DE) in all ration caused high blood triglycerides level. The result showed that the increase in fermented CP-JSC in the diets could have been due to decrease the blood triglycerides level although it was not differences among treatments. Fig. 2. Blood glucose levels of rabbits fed containing fermented CP-JSC as a SBM Substitution 4. Conclusion Based on feed consumption and blood metabolite profiles, it can be concluded that soybean meal can be substituted with protein concentrate from jatropha seed cake as a feed of post weaned Rex rabbits. However, more research is needed to study the effect of the use of jatropha protein concentrate on the production and reproduction performances of rabbit. Acknowledgements The authors express gratitude to Jenderal Soedirman University for funding the research trough DIPA BLU 2013. References [1] Makkar HPS, Francis G, Beckers K. Protein concentrate from Jatropha curcas screw-pressed seed cake and toxic and antinutritional factors in protein concentrate. Journal of the Science of Food and Agriculture. 2008.Volume 88, Number 9. p. 1542 1548 (7) [2] Saetae DT, Kleekayai, Jayasena V, Suntornsuk W. Functional properties of protein isolate obtained from physic nut (Jatropha curcas L.) seed cake. Food Science and Biotechnology. 2011.Vol 20 (1). p. 29 37. [3] Makkar HPS, Becker K. Challenges and opportunities for using byproducts from the production of biodiesel from jatropha oil as livestock feed. Proceedings of Animal Nutrition Association World Conference. 14 17 Feb 2009. New Delhi. India. p. 168 170. [4] Widiyastuti T, Sutardi TR, Indradji M. Optimalisasi protein concentrate bungkil biji jarak melalui kombinasi teknik kimiawi dan biologis menggunakan bal serta pemanfaatannya sebagai pakan ternak kelinci [Optimalitation of jatropha seed cake protein concentrate as rabbit feed through chemical and biological using lactic acid bacteria] Laporan Penelitian Riset Unggulan Tahun ke-1. Fakultas Peternakan, UNSOED. Purwokerto. 2012 [Bahasa Indonesia].
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