Available online at www.sciencedirect.

com

ScienceDirect
Energy Procedia 47 (2014) 290 – 294

Conference and Exhibition Indonesia Renewable Energy & Energy Conservation

[Indonesia EBTKE CONEX 2013]

Biorefinery of Jatropha Seed Cake by Lactid Acid Bacteria and the

Effects on Hematological Profile of Rex Rabbit
Titin Widiyastutia,*, Mohandas Indrajia, Agung Wibowob, Roy Hendrokoc
a
Animal Science Faculty of Jenderal Soedirman University, Jl. dr. Soeparno Karangwangkal, PO Box 110, Purwokerto 53123, Central Java.
b
PT JGE, Kudus, Central Java, Indonesia
c
PT. Sinarmas Agroresources and Technology, Sinarmas Land Plaza 2nd Tower 10th Floor, Jakarta 10350, Indonesia

Abstract

This research aimed: to increase the added value of Jatropha cultivation by using it as rabbit feed and to analyze the fermentation
of Jatropha seed cake by lactic acid and the effect on blood profile of post weaned Rex rabbits. The range and analysis of
variance of PCV, trombocyte, TPP and fibrinogen are presented in these documents. Conclusion, to increase the added value of
Jatropha cultivation is by utilizing the seed cake as complete feed for rabbits up to 12% level without affecting the blood profile
of post weaned Rex rabbits.

© 2014
© 2014The
TheAuthors.
Authors.Published
Published
by by Elsevier
Elsevier Ltd.Ltd.
Selectionand
Selection andpeer-review
peer-review under
under responsibility
responsibility of Scientific
of the the Scientific Committee
Committee of Indonesia
of Indonesia EBTKEEBTKE Conex 2013.
Conex 2013

Keywords: Jatropha curcas; complete feed; rex rabbit; packet cell volume; platelet count; total plasma protein and fibrinogen
____________________________________________________________________________________________________________________

1. Introduction

High fuel oil consumption in Indonesia has led to the ever increasing biofuel exploration, resulting in by-product
excess that is subject to utilize zero waste products. Jatropha curcas is one of biofuel sources that one ton dried
Jatropha seeds produce 200-300 liter oil and 700-800 kg seed cake waste [1]. Shell less of seed cake contains 56.4%
crude protein, higher than soybean meal 48%. Rich nutrition in Jatropha seed cake is a significantly potential feed
utilization; the biological value however is not equally high because of antinutrients such as phorbolesters, tannins,
phytates, saponins, trypsin inhibitor, and lectins that inhibit the utilization of Jatropha seed cake as feed.
Detoxification methods have been administered to eliminate the negative effect of the antinutrients by heating and
____________________

___________________________
* Corresponding author. Tel.: +628164288555; fax: +62-281-638792.
E-mail address: dyast72@yahoo.com

1876-6102 © 2014 The Authors. Published by Elsevier Ltd.

Selection and peer-review under responsibility of the Scientific Committee of Indonesia EBTKE Conex 2013
doi:10.1016/j.egypro.2014.01.227
 Titin Widiyastuti et al. / Energy Procedia 47 (2014) 290 – 294 291

chemical treatments [2, 3, 4], acid base precipitation technique [5], fermentation with Marasmius spp [6],
fermentation with lactic acid bacteria (Lactobacillus spp dan Bifidobacter spp) and saccharides addition [7]
followed by FOS (Fructooligosccharides) addition up to 1.5 % in layer and broiler feeds [8] Meat self-sufficiency
still depends mostly on ruminant while non-ruminant commodity is well develop yet. Diversification in non-
ruminant products can significantly contribute to meat self-sufficiency plan in 2014. One of meat producing
commodity is rabbits, and Rex rabbits are protein source as nutrition fulfilment alternative in society. Rabbits are
potential to produce meat and pelt and as decorative rabbits [9]. High quality feed will support a fast rabbit meat
production concerned to a relatively short production period (meat and population) compared to ruminants.
Providing high quality feed and no competition with other cattle are required as well as the importance of exploring
non-conventional feed sources. Optimizing biorefinery of Jatropha seed cake therefore needs to accomplish. The
study of its application as extensive feed to various poultry both ruminants and non-ruminants is of good possibility.
It is therefore important to observe the effect of Jatropha seed cake on haematological profile of rabbits (packed cell
volume, red blood cell, fibrinogen and total plasma protein). Protein-contained feed affects the blood profile of
cattle, so blood profile in Rex rabbits is a significant indicator to determine whether feed containing Jatropha seed
cake affects the rabbits health status.

2. Research methods

This research administered 21 post weaning Rex rabbits of 120 days old, fermented Jatropha curcas seed cake
and feed (iso protein and iso energy) ration formula is presented in Table1. Materials and utility used in the research:
J. curcas seed cake, inoculation device and Bifidobacterium spp.culture, MRS (de Man Rogosa Sharpe) broth
media, molases, pasteured milk, sterilized aquades, bunsen, plastic covers, rubber band, glass bottle. Pellet machine,
feed and water container, sensitive balance scale of 10 x 10-3 kg and 5 kg scale, cage thermometer, disinfectant,
syringe and Na-EDTA (Natrium Ethylen Diamin Tetra Acetic Acid). Completely Randomized Design with three
treatments and seven repetitions was applied. The treatments consist of R0 (complete pellet without fermented
Jatropha curcas seed cake/ control, R1 (complete pellet with 6 % fermented J. curcas seed cake), R2 (complete
pellet with 12 % fermented J. curcas seed cake). Fermentation of J.curcas seed cake used Bifidobacterium spp
culture for 15 days at 37° C, and the fermented product was used to compose complete pelleted feed. Rabbits were
reared for 30 days, data of feed consumption and body weighing were taken once a week, and intracardiac blood
collection was on the 30th day after the rabbits underwent 5 hour fasting. The measured variables were packed cell
volume (%) and thrombocytecount (per mm3), total plasma protein, fibrinogen and feed consumption as supporting
variable. Data collected were subject to variance analysis continued by orthogonal polynomials [10].

Table 1. Nutrient content of feed.

Feed R0 R1 R2
(0%) (6%) (12%)
Nutrients (%)
Water 11.45 9.70 8.04
Ash content 7.78 8.52 8.38
Crude protein 19.30 18.42 18.66
Crude fat 7.89 8.41 9.48
Crude fibre 18.27 18.08 18.16
BETN 46.76 46.57 45.32
Ca 1.31 1.26 1.36
P 0.87 0.91 1.03
GE (kkal/kg) 2,873.690 2,757.234 2,740.679
Note:
R0= Complete pelleted feed without fermented Jatropha seed cake (control)
R1= Complete pelleted feed with 6% fermented Jatropha seed cake
R2= Complete pelleted feed with 12 % fermented Jatropha seed cake
292 Titin Widiyastuti et al. / Energy Procedia 47 (2014) 290 – 294

3. Result and discussion

3.1. Consumption

Result showed daily feed consumption of Rex rabbits was 61.47 to 85.01 g/head/day with declining consumption
tendency in every increasing level of fermented Jatropha seed cake in complete pelleted feed. Variance analysis
result performed treatments significant effect (P < 0.01) on feed consumption. The declining feed consumption of
treatment R2 in Rex rabbits was assumed as the result of falvour of fermented Jatropha seed cake that
dominated the flavour of complete feed pellet composition if compared to control feed that was consumed more.
But, the rabbit consumption is on normal level (65 – 85g kg 1 body weigt) [11]. Rabbits palatability was expected to
improve by eliminating the flavour and providing longer adaptation period [12].

90
80
70
Consumption (g/d/head)

60
50
40 Y = 83 . 6885 - 1 . 9619 X
r 2 = 43 . 45 %
30
20
10
0
0 2 4 6 8 10 12 14

Fermented Jatropha seed cake (%)

Fig. 1. Feed consumption of rabbits that received feed fermented Jatropha seed cake.

3.2. Packed cell volume (PCV)

PCV is a measurement used to detect anaemia or polycythemia and to predict the change of hemodilution or
homo concentration. PCV is used with red blood cell in calculation of mean cell volume (MCV) and with
haemoglobin and corpuscular (MCHC). The result showed the average packed volume cell of R 0, R1 and R2 was
31%, 32.28 % and 28.42%, respectively, indicating the highest average of packed cell volume in treatment R 1and
the lowest in treatment R2. Estimated PCV in rabbits was 36% to 48% [13], and 37.67% in local rabbits [14]. Low
PCV average in research result was due to the low feed consumption. Variance analysis result indicated feed
treatment containing fermented Jatropha seed cake did not significantly affect Packed Volume Cell (P > 0.05). No
significant difference of PVC in treatments indicated that fermented Jatropha seed cake could be utilized to rabbits
feed up to 12%. The factors influencing PVC were age, race, sex, and long duration of transporting sample from
storage to testing site during which the blood cells lysis took place [15]. Low PVC in this research did not induce
anemia in experimental rabbits because 12% fermented Jatropha seed cake did not cause illness to the rabbits. It was
likely because the Jatropha seed cake fermented using Bifidobacterium spp. could lower hemagglutinin in Jatropha
seed cake as the main factor of red blood cell damage leading to anemia in rabbits.

3.3. Platelet/ thrombocyte count

Research result showed the average platelet count in control (R0), R1 and R2 was 69,428.6 (/mm3), 73,000
(/mm3) and 61,285.7 (/mm3), respectively. Analysis of variance indicated 12% fermented Jatropha seed cake did not
significantly affect platelet count in post weaned Rex rabbits P > 0.05). Normal platelet count in rabbits is less than
 Titin Widiyastuti et al. / Energy Procedia 47 (2014) 290 – 294 293

150,000/mm³. Thrombocytopenia is one of side effects of sulfonylurea because it prevents megakaryocyte –the giant
cell in bone marrow. This cell forms platelet and exert a small amount of cytoplasm into bloodstream. One
megakaryocyte can form 4,000 platelets [16]. Treatment R1 had higher platelet than that of R0 and R2 and was still
considered normal since subnormal platelets was proven that lectineantinutrient in Jatropha seed cake could be
neutralized through Bifidobacterium spp. fermentation as observed from zero mortality and sickness in rabbits.
Antinutrient in Jatropha seed cake belongs to the group of lectin, antitrypsin (trypsin inhibitor), saponin and
porbholester. Porbholester, if consumed can cause death in cattle [17]. Protease inhibitor and lectin can disturb
enzymatic digestion digestive system; moreover, lectin can attach to and damage intestinal epithelial cell receptor
that eventually causes digestion and nutrient absorption disorders [18]. [8] further reported lectin in Jatropha seed
cake can be the host of lactic acid in chicken intestine.

3.4. Total plasma protein (TPP)

The average total plasma protein of Rex rabbits ranged from 4.92 – 5.09 g/dL, with increasing tendency linear to
the increase of fermented Jatropha seed cake in complete feed. Analysis of variance showed no treatment effect on
total plasma protein (P.0.05), indicating complete feed pellet R0, R1, and R2 did not result in different plasma protein
of post weaned Rex rabbits. Rabbits with R0 treatment in fact performed a slightly subnormal TPP compared to that
of R1 and R2. Higher feed consumption in R0 than that of R1 and R2 suggested higher biological value in protein
content of R1 and R2, and protein in Jatropha seed cake could substitute protein in copra meal or soybean meal. In
accordance with [19], plasma protein content was affected by protein content in feed, sex and growth rate. Result
showed linear increase of total plasma protein to that of Jatropha seed cake (not statistically significantly different).
During experiment however, no ill rabbits biologically indicated that R 2, complete pellet feed containing 12%
fermented Jatropha seed cake, was still tolerable because it was within standard total plasma protein count: 5.0 – 8.0
g/dL [20]. Plasma protein increases when body resists infection through immunoglobulin system. Plasma protein
concentration will increase to respond hypergammaglobulinemiasymptom-disease [21].

3.5. Fibrinogen

The average fibrinogen in Rex rabbits was 0.74 – 1.52 g/dL, with tendency of linear increase to that of fermented
Jatropha seed cake level in complete pelleted feed Analysis of variance indicated treatments did not affect
fibrinogen. (P > 0.05). Fibrinogen of Rex rabbits in treatment R2 was beyond normal – 1.52 g/dL although it was
not statistically significantly different while [22] stated that standard fibrinogen level was 0.3 – 0.9 g/dL. It indicated
thelectin content in fermented Jatropha seed cake given at 12% level, accordingly homeostatis was absent in Rex
rabbits’ physiology but did not cause illness or death in rabbits. Theoretically, fibrinogen increase indicates general
health disorder, trauma and infection. Fibrinogen is the essential component in blood clotting (coagulation).
Coagulation takes place when cattle suffer from suppurating sore and acute or chronic disease. Fibrinogen level
declines in liver disease.

Table 2. Hematological profile of Rex rabbits with Jatropha seed cake treatment.
Treatments PCV (%) Thrombocyte (/mm3) Total Plasma Protein (g/dl) Fibrinogen (g/dl)
R0 31.00 ± 6.85 69428.6 ± 0.128 4.92 0.74

R1 32.28 ± 5.02 73000.0 ± 0.094 5.02 0.98

R2 28.42 ± 3.30 61285.7 ± 0.144 5.09 1.52

294 Titin Widiyastuti et al. / Energy Procedia 47 (2014) 290 – 294

4. Conclusion

The added value of Jatropha cultivation can be increased by optimizing Jatropha seed cake as complete feed.
Composition up to 12% fermented jatropha seed cake in the rabbit feed, has no effect on the blood profile (packet
cell volume, platelet count, total plasma protein and fibrinogen) of post weaned Rex rabbits.

Acknowledgements

The authors expers gratitude to Jenderal Soedirman University for funding the research, and to Muamar
Riza Sabara and Rommi Prima Satya as enumerator.

References

[1] Brodjonegoro TP, Rexowardjojo IK, Tatang, Soerawidjaja H. Jarak pagar sang primadona. Departemen Teknik Kimia, Lab. Termofluida
dan Sistem Utilitas, Kelompok Riset Biodiesel ITB. Available from http://www.pikiran-rakyat.com/cetak/
2005/1005/13/cakrawala/utama02.htm. 2005, accesed at November 24th 2011.
[2] Aregheore EM, Becker K, Makkar HPS. Detoxification of a toxic variety of Jatropha curcas using heat and chemical treatments, and
preliminary nutritional evaluation with rats. S.Pac.J.Nat.Sci. 2003; 21:50–56.
[3] Herera MJ, Siddhuraju P, Francis G, Ortiz GD, Becker K. Chemical composition, toxic/ antimetabolic constituens, and effects of different
treatments on their levels, in four provenances of Jatropha curcas L. from Mexico. Food Chem. 2006; 96:80-89.
[4] Chivandi E, Erlwanger KH, Makuza SM, Read JS, Mtimuni JP. Effects of dietary Jatropha curcas meal on percent packet cell volume,
serum glucose, cholesterol and triglyceride concentration and alpha- amylase activity of weaned fattening pigs. Research Journal of Anim.
Vet. Sci 2006; 1(1):18-24.
[5] 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; 88(9):1542-1548.
[6] Irawan A. Kadar lignin dan serat kasar bungkil biji jarak (Jatropha curcas) yang difermentasi dengan Jamur Marasmius sp. Skripsi.
Fakultas Peternakan Unsoed. Puwokerto; 2007.
[7] Widiyastuti T, Prayitno CH. Manipulasi lingkungan mikrointestinal menggunakan lectin bungkil biji jarak sebagai media perlekatan
bakteri asam laktat serta pengaruhnya terhadap performans unggas. Laporan Penelitian Hibah Bersaing XV. Tahun ke-1; 2009.
[8] Widiyastuti T, Prayitno CH, Iriyanti N. Manipulasi lingkungan mikrointestinal menggunakan lectin bungkil biji jarak sebagai media
perlekatan bakteri asam laktat serta pengaruhnya terhadap performans unggas. Laporan Penelitian Hibah Bersaing XV. Tahun ke-2; 2010.
[9] Priyono. Melirik potensi kelinci; 2009. Available from http:/www.ilmupeternakan.co.cc/blogger/archive html, accessed at Februari 10th
2011.
[10] Steel RG, Torie JH. The principle and procedure of statistic. Jakarta: Gramedia Pustaka Utama; 1993. Diterjemahkan oleh Sumantri B.
Prinsip dan prosedur statistika; 1991.
[11] de Blass C, Weisman J. The nutrition of the rabbit. Wallingford, UK: CABI Publishing; 1998.
[12] Widiyastuti T, Indraji M. Kajian complete feed berbahan dasar bungkil biji jarak terfermentasi ditinjau dari performans kelinci lepas sapih.
In: Prosiding Seminar Nasional Pengembangan Sumberdaya Pedesaan dan Kearifan Lokal Berkelanjutan. LPPM Unsoed. Purwokerto.
Nopember 23-24th 2011. ISBN: 978-979-9204-51-6; 2011.
[13] Harkness JE, Wargner JE. The biology and medicine of rabbits and rodents. 3rd ed. In: Lea, Fibiger, editors. Philadelphia. London; 1983.
[14] Hana, A., Sumartono, Airin CM, Astuti P. Pengaruh Eimeria magna Terhadap Kondisi Fisiologik Kelinci Lokal. J.Sain Vet. 2010; 28(2):
89–97.
[15] Hohenhaus, Ann E. Transfusions containing red blood cells. In: Proceeding of the WSAVA Congress in Australia; 2007.
[16] Triana E, Nurhidayat N. Pengaruh pemberian beras yang difermentasi oleh Monascus purpureus JmbA terhadap darah tikus putih (Rattus
Sp.) hiperkolesterolemia. Bogor: Balai Penelitian Mikrobiologi, Pusat Penelitian Biologi, Lembaga Ilmu Pengetahuan Indonesia (LIPI);
2006.
[17] Pasaribu T, Wina E, Tangendjaja B, Iskandar S. Performans ayam yang diberi bungkil biji jarak pagar (jatropa curcas) hasil olahan secara
fisik dan kimiawi. JITV 2009; 14:11-18.
[18] Huisman J, Jansman AJM. Dietary effects and some analytical aspects of antinutritional factors in peas (Pisum sativum), common beans
(Phaseolus vulgaris) and soyabeans (Glycine max L.) in monogastric farm animals. A literature review. Nutrition abstracts and reviews.
Series B. Livestock feeds and feeding 1991; 61:901-921.
[19] Grimminger FJ. Threonine deaminase from a nonsense mutant of Escherichia coli requiring isoleucine or pyridoxine: evidence for half-
of-the-sites reactivity. J Bacteriol. 1976; 126(1): 100-7.
[20] Laboratorium Phatologi Klinik. Petunjuk praktikum phatologi klinik. Jogjakarta: Fakultas Kedokteran Hewan, Universitas Gadjah Mada;
1998.
[21] Schalm OW, Jain NC, Carroll EJ. Veterinary hematology. 5th ed. In: Lea, Febiger, editors. Philadelphia; 2000.
[22] Jain NC, Febiger LE. Essentials of veterinary hematology. Philadelphia; 1993. p. 349-353.