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Effect of Solid State Fermentation and Other Processes on the Lathyrus Sativus (kheshari Dal) Toxin by Chick Bioassay

A. B. M. Moslehuddin
Institute of Nutrition and Food Science, University of Dhaka, Dhaka, Bangladesh

M. A. Malek
Institute of Nutrition and Food Science, University of Dhaka, Dhaka, Bangladesh

Abstract/ Executive Summary:

Lathyrus sativus seeds were processed by different methods for removal or destruction of their natural toxins. Toxins were extracted from the seeds, partially purified and injected intraperitonally into one day old male chicks as a sensitive bioassay for th effectiveness of the processed seeds. The chicks showed typical neurological symptoms such as head retraction, neck bending after injection of the concentrated extracts. Similar symptoms were observed in chicks injected with the extracts of seeds soaked overnight in water or in saturated lime followed by steaming, autoclaving and fermenting at 30°C for 30 hours. Toxicity symptoms were observed to all groups of the experiment except 1 and 6. Extracts from soaked overnight, washed five times showed 75% and two times showed symptom 75% and 100%. Soaked 5 times and fermented for 30 hours showed symptom 0.00% and control containing sterile distilled water showed 0.00% symptoms. Washing twice and five times have some effect on reducing toxicity in the seeds and solid state fermentation also showed some reduction of toxicity from seeds. Lime water treatment and autoclaving appeared to have little effect on toxin removing or destroying.

Keywords: Lathyrus satirus, Chick Bioassay, Toxin, Kheshari dal

Location: Institute of Nutrition and Food Science, University of Dhaka, Dhaka, Bangladesh

Start Date:

End Date:

Program Area: Risk Management in Agriculture

Commodity: Grasspea

Objectives:

To process Lathyrus sativus seeds by various methods for neurotoxin removal or destruction using less concentrated toxin (0.5 ml) instead the sensitive chick of 1 ml and bioassay was done to evaluate the effectiveness of these processing treatments.

Methodology:

Fermentation and extraction of Toxins from the treated seeds. Sample of Lathyrus sativus seeds were obtained from the local market. After receiving the seeds, undesirable matters were removed manually. The seeds were than dehulled by burr mill in dry condition so that molds cannot grow on the dehulled seeds. After removing the seeds coat, the seeds were kept at room temperature. At the regular periods the seeds were examined for the presence of moldy or damaged seeds. Before fermentation, the seeds were soaked in water overnight (seeds and water ration being 1:3). After soaking water was drained off, washed and cleaned. Preparation of Tempeh (solid state fermented product) was followed by the modified method. The seeds were then steamed for 10 minutes to remove some of the Microflora. Then the seeds were dried at oven for removal of excess water for one hour. After removal of excess water the seeds were inoculated with 0.5% inoculum of Rhizopus oligosporus (Tempeh inoculum) obtained from the Nutrition Research Development Centre at Bogor, Indonesia. The inoculated seeds were packed in petridishes and incubated at 30°C for 24 hours. After incubation, the fermented seeds were kept at 0°C. The raw dehulled seeds (50 gram portions) were powdered and blended with 300 ml of distilled water for 10 minutes. Extractions of mixture were made by heating and stirring for 15 hours. After extraction, the mixture was centrifuged at 3,000 rpm for 20 minutes in a centrifuged and the supernatant was decanted. The residue was extracted again and combined with the first extraction after recantation. The decanted supernatant was then concentrated to 12.5 ml for an appropriate toxin concentration of 20 mg/ml by rotatory evaporator (E Corning, England). The concentrated raw seed extracts was then adjusted to p 6.8 with 40% NaOH. A second batch of the seeds was soaked with 150 ml of water overnight, washed twice, steamed for 10 minutes and extracted as above. A third batch of the seeds was soaked and washed twice and steamed for 10 minutes and inoculated with 0.5% tempeh inoculum CRhizopus oligosporus). A 4th batch of seeds was soaked overnight and washed five times, extracted as before. A fifth batch of seeds was soaked overnight and washed five times and inoculated with the incluum of Rhizopus oligosporus Packed with petridish and inoculated for 30 hours at 30°C. The sixth batch of seeds was soaked in saturated lime water and was extracted as before. The seventh batch of seeds was soaked in saturated lime water and autoclaved for 10 minutes, was extracted as before. The lime was purchased from local market of Dhaka. All the seeds of different batches were extracted and concentrated as before and kept for Bioassay of chicks. Seeds of all th batches were extracted by the same procedure to same volume. For toxicological evaluation of the seven seeds extracts from different processing procedures, a one day old chick bioassay was accomplished. Male white chick weighing 30-35 gms were obtained from the Department of Poultry, Mirpur, Dhaka. The chicks were divided into eight groups and were individually ip injected with 0.5 ml of the seed extracts. Each chick in group one was injected with 0.5 ml of sterile water (Control) by (ip) injection. Group two received extracts of raw seeds. Group three received extracts of soaked overnight and washed twice. Group four received extracts of soaked overnight, washed twice and fermented with Rhizopus oligosporus for 30 hours at 30°C. Group five received extracts of soaked overnight and washed five times. Group six received extracts of soakd overnight and washed five times and fermented as before. Group seven received extracts of lime water soaked seeds for overnight. Group eight received extracts of lime water soaked an autoclaved seeds. All samples after soaking was steamed for 10 minutes. Chicks were maintained on chick starter (Dhaka, Bangladesh) Placed in an electrically heated room at 35°C.

Source of Information: Bangladesh J. Nutr. Vol. 8. Nos. 1 & 2, June 1995, ISSN 1013-6037

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Treating chicks with the extracts of seeds soaked overnight and washed 5 times showed symptom 75% within 30 minutes and 3 hours. Treating chicks with the extracts of seeds soaked overnight washed five times and fermented showed 0.00% symptom of toxicity within 30 minutes and 3 hours. Treating chicks with extracts from soaked overnight with saturated lime water showed symptom of neurotoxicity 75% within 30 minutes and 100% within 3 hours. Chicks treated with the extracts of seeds from lime water soaked and autoclaved showed 100% symptom of neurotoxicity in both in 30 minutes and 3 hours.

Knowledge Management: Journal