Comparative evaluation of toxic and non-toxic Jatropha genotypes

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Comparative evaluation of toxic and non-toxic Jatropha genotypes Harinder Makkar

Variation in seed numbers per fruit from toxic and non-toxic Jatropha curcas 2 Non-toxic variety Percentage of fruits with: 1 seed 2 seeds 3 seeds 4 seeds Shrub 1 12,0 26.5 55.5 6.0 Shrub 2 8.0 19.1 64.9 8.1 Shrub 3 4.4 24.5 54.5 16.6 Average 8.1 23.4 58.3 10.2 Toxic-variety Shrub 1 13.7 27.5 53.6 5.2 Shrub 2 14.4 22.4 56.8 6.4 Shrub 3 15.8 27.3 45.6 11.3 Average 14.6 25.8 52.0 7.6

Variation in physical parameters of seeds and in oil content of seeds and kernels from toxic and non-toxic Jatropha curcas (n = 60; 20 from 1 tree) 3 Toxic variety Average of fruits with: Seed mass (g) Shell mass (g) Kernel mass (g) Shell/Seed Mass (%) Oil Kernel (%) Oil Seed (%) Kernel protein (%) 1 seed 0,77 0,29 0,47 37,7 56,5 34,8 27.1 2 seeds 0,78 0,30 0,49 38,5 57,2 36,0 26.3 3 seeds 0,80 0,31 0,49 38,7 57,4 35,3 26.6 4 seeds 0,77 0,29 0,48 37,7 56,9 35,5 26.8 Average 0,78 0,30 0,48 38,0 57,0 35,4 26.7 Non-toxic variety Average of fruits with: Seed mass (g) Shell mass (g) Kernel mass (g) Shell/Seed Mass (%) Oil Kernel (%) Oil Seed (%) Kernel protein (%) 1 seed 0,73 0,27 0,46 37,0 55,5 35,3 28.5 2 seeds 0,73 0,26 0,47 35,6 57,6 37,0 26.9 3 seeds 0,72 0,26 0,47 36,1 57,5 37,1 26.9 4 seeds 0,72 0,25 0,45 34,8 56,2 36,1 26.4 Average 0,72 0,26 0,46 35,9 56,7 36,4 27.2

Composition of seed meal from toxic and non-toxic Jatropha curcas 4 Toxic variety Non-toxic variety Crude protein (%) 56.4 63.8 Lipid (%) 1.5 1.0 Ash (%) 9.6 9.8 Gross energy (MJ/kg) 18.2 18.0 Neutral Detergent Fibre (%) 9.0 9.1 Harinder Makkar

5 Chemical composition of capsules and shell of Jatropha curcas Constituents (% DM) Toxic variety Non-toxic variety Capsule husk Shell Capsule husk Shell Crude protein 6.4* 4.3-4.4 Lipid 0.6* 0.7-0.5 Ash 17.9* 6.0-2.8 Neutral detergent fibre (NDF) Acid detergent fibre (ADF) 65.9** 83.9-89.4 61.3** 74.6-78.3 Acid detergent lignin 14.4** 45.1-45.6 Gross energy (MJ Kg -1 ) 14.5* 19.3-19.5 * from the hot desert of Egypt ** from Ife, Nigeria Harinder Makkar

6 Potentially useful fractions of Jatropha Seeds 1000 kg Kernel 625 kg Fruits 1600 kg Husk 600 kg Shell 375 kg Capsule sheath: Mulch, fuel briquettes (14.5 MJ/kg) Seed shell: high energy content (18MJ/kg), ideal for process energy down the line, production of briquettes for sale Potential products

7 Non-toxic similar to the toxic one except for the phorbol esters Important antinutrients in Jatropha seed meal (dry matter basis) Component Toxic Non-toxic Phorbolesters (mg/g kernel) Total phenols (% tannic acid equivalent) Tannins (% tannic acid equivalent) Phytates (% dry matter) Saponins (% diosgenin equivalent) Trypsin inhibitor (mg trypsin inhibited per g sample) Lectins (1/mg of meal that produced haemagglutination per ml of assay medium) ND: not detected 2.79 0.36 0.04 9.40 2.60 21.3 102 ND to 0.11 0.22 0.02 8.90 3.40 26.5 51 Harinder Makkar

Essential aminoacid composition (g/16gn) of Jatropha seed meal compared to soybean and FAO child ref. protein 8 Aminoacids Toxic Non-toxic Soybean meal FAO (ref protein) Methionine 1.91 1.76 1.22 Cystine 2.24 1.58 1.70 2.50 Valine 5.19 5.30 4.59 3.50 Isoleucine 4.53 4.85 4.62 2.80 Leucine 6.94 7.50 7.72 6.60 Phenylalanine 4.34 4.89 4.84 Tyrosine 2.99 3.78 3.39 6.30 Histidine 3.30 3.08 2.50 1.90 Lysine 4.28 3.40 6.08 5.80 Arginine 11.8 12.9 7.13 Threonine 3.96 3.59 3.76 3.40 Tryptophan 1.31-1.24 1.10 Harinder Makkar

Nutritional parameters of Jatropha seed meals compared to soybean meal 9 Toxic Non-toxic Soybean meal Digestible organic matter (%) 78 77.3 87.9 Metabolizable energy (MJ kg -1 DM) 10.9 10.7 13.3 24 h in vitro rumen degradable nitrogen (% of total N) 43.3 28.9 80.9 The current plantation efforts exclusively use the toxic provenance Higher productivity outside Mexico Higher tolerance to diseases Harinder Makkar

10 The seed cake from the toxic Jatropha current uses Bio fertilizer cum bio pesticide Contains 5.7 6.5% N, 2.6 3.0% P2O5, 0.9 1.0% K2O, 0.6 0.7% CaO and 1.3 1.4% MgO The phorbol esters contained acts as a pesticide against harmful soil organisms Literature suggests that the phorbol esters are completely destroyed after 6 days validation required The limited quantity of the seed cake produced currently enjoys high demand for this purpose in some Indian states As a substrate for production of biogas Use as direct fuel in power generators Jatropha seed cake

The seed kernel meal potential as animal feed ingredient 11 Average gross chemical composition of Jatropha kernel meal Crude Protein % Lipid % Ash % Gross energy (MJ/kg) Neutral detergent fibre % 60-63 < 1.0 10 18.0 9 Every kilo of oil extracted yields about 0.75 kilo of seed meal if kernels are solvent extracted Soybean meal with 45% crude protein content has an international market price 300US$/ton for use as animal feed Based on crude protein content the Jatropha meal could have a price to the order of 430 US$/ton if detoxified The seed meal from the toxic Jatropha contains a variety of antinutrients and one toxic substance the phorbol esters: it needs to be detoxified before use as animal feed ingredient JATROSOLUTIONS Jatropha seed GMBH cake

Raw meal of the non-toxic Jatropha not suitable as such 12 Lower growth noticed in fish and rats Chemical and physical processing required to remove the secondary compounds (and increase protein digestibility?) Treatments devised - moist heating at 120 C helped remove trypsin inhibitors and lectins - addition of microbial phytase (500 FTU) resulted in mitigation of adverse effects of phytate

The potential of seed kernel meal as fish feed ingredient established using treated meal from the non-toxic provenance 13 Percentage body weight gain of a common culture fish (carp) fed experimental diets containing 50 or 75% of protein supply from Jatropha meal or soybean meal for 8 weeks 700 600 % BWG 500 400 300 200 100 0 control Jatropha meal J. meal+ Lysine 500 50% 75% 400 J. meal + phytase % body weight gain 300 200 100 0 Control Soyabean meal Jatropha meal The threshold level at which phorbol esters caused adverse effects was 15 ppm (15 µg/g) in the diet

Even rumen bacteria were unable to break down phorbol esters in the seed meal 14 Phorbol ester (PE) content of residues from Jatropha seed cake or meal (with and without shell) after 24 h in buffered rumen liquor Sample Jatropha meal (defatted) Jatropha seed cake with shell and oil residues Jatropha seed cake with shell, defatted PE content (mg/g) before incubation 1.67 ± 00 0.63 ± 00 0.58 ± 00 PE content (mg/g) after incubation 1.69 ± 0.08 0.55 ± 0.06 0.40 ± 0.08 Hence the meal without removal of phorbol esters cannot be used as feed ingredient for ruminants

Detoxification of seed meal successfully achieved at laboratory scale 15 Body weight (g) 18 16 14 12 10 8 6 4 2 0 Control Non-toxic Jatropha diet Detoxified Jatropha diet 0 10 20 30 40 50 60 Days The detoxification method is scalable and can be integrated with oil extraction Validation of upscaling and nutritional quality through long term field tests required Body weight development of common carp fed diets containing 75% Jatropha meal compared to control Jatropha seed cake

16 Some of our ongoing studies Jatropha toxins in the biodiesel production chain Distribution of phorbol esters in different parts of Jatropha plant Preparation and use of Jatropha protein concentrate Long-term feeding studies using detoxified seed meal Upscaling of the Jatropha seed meal detoxification process Development of efficient de-shelling technology Optimization of screw press technology for extraction of oil from kernels and kernels containing different amounts of shells Development of a cooking stove for use with Jatropha oil Development of small scale combustion systems (furnace) for using shells as energy source Economic evaluation of the detoxification process, and its accounting in the overall economic assessment of Jatropha-based small and large scale oil production systems Socio-economic evaluation of Jatropha cultivation and use Harinder Makkar

17 Future Research Needs SHORT TERM Generation of value-added products in the process of biodiesel production from Jatropha oil, in particular detoxification of seed meal and isolation of phorbol esters Evaluation of existing genotypes (both toxic and non-toxic) under different environmental conditions and selection of genotypes for different conditions (focus: degraded lands) Optimization of agronomic practices (at least 1 ton oil/ha/year) Optimization of mechanical harvesting of Jatropha seeds Optimization of tissue-culture procedure and establishment of protocols for genetic transformation Researchable issues (1/3)

18 Future Research Needs MEDIUM TO LONG TERM Genetic improvement of Jatropha through conventional, mutation, and/or molecular breeding Cold/Frost resistance High oil yield and change of fatty acid composition Early maturity and seed productivity High efficiency of water and nitrogen use Fate of Jatropha toxins in the biodiesel production chain and in the environment and their impact on human health and environment Integration of Jatropha with leguminous food-feed crops and its socioeconomic impact Researchable issues (2/3)

19 Future Research Needs Development of criteria for certification of seeds Evaluation of sustainability of Jatropha as an energy source (life cycle analysis) Models for quantification of the tradable emission credits, both through sequestration of CO2 in plant biomass and through the use of the CO2 neutral bio-diesel produced Socio-economic analysis of Jatropha cultivation and use Researchable issues (3/3)

20 ACKNOWLEDGEMENT Prof. Dr. K. Becker and Dr. George Francis for useful discussions and inputs

21 Further information can be obtained from: Prof. Dr. Harinder Makkar Project Leader: BMBF-MOST Jatropha Project Institute for Animal Production in the Tropics and Subtropics (480b), Stuttgart E-mail:makkar@uni-hohenheim.de

22 A request The data in this presentation/document is from our laboratory. While using data from this document, please quote reference from literature. If you do not find the original document/reference, please contact us. Thanks in anticipation of giving credit where it is due.