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Detection of Aflatoxin in Poultry Feed and Feed Materials Through Immuno Based Assay from Different Poultry Farms and Feed Factories in Bangladesh

Mahbuba Akter Lubna*
Quality Control Department, Akij food & Beverage Ltd., Krisnapura, Dhamrai, Dhaka, Bangladesh

Mita Debnath
Kazi Farms Poultry Laboratory, Shafiqul Villa, Chandana, Chowrasta, Gazipur, Bangladesh

Farzana Hossaini
Infectious Diseases Division, International Centre for Diarrhoeal Disease Research Bangladesh, Dhaka, Bangladesh.

Abstract/ Executive Summary:

Current study investigated the occurrence of aflatoxin contamination in poultry feed and feed materials in different poultry farms and feed factories in Bangladesh. A total of 100 samples of finished feed and raw feed materials were collected and tested through direct competitive Enzyme-Linked Immunosorbent Assay (ELISA) for total aflatoxin detection. Overall, 97% samples (n=97/100) in our study, were found positive for aflatoxin contamination. Among finished feed categories, layer grower feed contained highest level of aflatoxin with a mean value of 21.64 ppb whereas layer feed was less susceptible for aflatoxin contamination (mean value 9.49 ppb). Between raw feed materials, maize samples were highly contaminated (n=15/15, 100%) with aflatoxin while 86.67% soybean samples showed positive result. Twenty one percent (21%) of the samples in our study contained aflatoxin concentration more than the acceptable limit employed by USFDA and many other countries which might pose severe health risk to poultry and human consumer. Proper surveillance and immediate control measures should be taken to ensure safe poultry feed and feed materials.

Keywords: Aflatoxin, Poultry Feed, Enzyme-Linked Immunosorbent Assay (ELISA)

Location: Kazi Farms Poultry Laboratory, Gazipur, Bangladesh

Start Date: 00-07-2017

End Date: 00-02-2018

Program Area: Risk Management in Agriculture

Commodity: Feed, Health hazard

Objectives:

Present study was designed to explore the occurrence of aflatoxin contamination in poultry feed and feedstuffs available in Bangladesh.

Methodology:

Sample Collection: A total of 100 poultry feed samples including finished feed (n=70) and raw feed materials (n=30) were collected from different poultry farms and feed production factories in Bangladesh during eight months period from July, 2017 to February, 2018 and transferred immediately to laboratory for testing. All test procedures were performed in Kazi Farms Poultry Laboratory, Gazipur, Bangladesh. The finish feed samples represented five categories, namely, broiler starter feed (n=13), broiler grower feed (n=18), broiler finisher feed (n=12), layer feed (n=12), and layer grower feed (n=15) while raw feed materials represented two categories including maize (n=15) and soybean (n=15). In order to achieve reasonably representative result approximately 500g of each sample was collected from poultry farms and feed production factories with proper labeling including date, factory name, sampling place and time of collection. All the samples were homogenized and thoroughly ground for analysis. Sample Preparation: The sample was prepared according to the previous study performed by Kehinde et. al. (2014) in Ogun State, Nigeria 21 . Dilution was done into two different ways. One is 5 fold dilution in which ten grams (10g) of feed sample or feed ingredient were taken in conical flasks and labelled according to the sampling location. Fifty milliliter (50 ml) of 70% methanol was added into each flask to extract the aflatoxin in the feed samples. Another dilution was done by following 10 fold dilution in which Five grams (5 g) of feed or ingredients were taken and fifty milliliter (50 ml) of 70% methanol was added into each flak to extract the aflatoxin. After few minutes the samples were filtrated by using Millipore filtrate paper into specific labelled test tubes. Quantification of Total Aflatoxin in Feed Sample: The concentration of aflatoxin was analyzed by a direct competitive Enzyme-Linked Immunosorbent Assay (ELISA), which is based on antigen antibody reaction 22. For the enumeration, Agra Quant® Total Aflatoxin Assay 4/40 Kits (Romer Laboratory Inc. Singapore) was used. Extracted samples were diluted into dilution wells by adding 200 ml of conjugate solution (provided with the kit) with 100 ml of extracted samples. After proper mixing, 100 ml conjugate mixed extract samples were transferred into antibody coated wells and incubated for 15 min at room temperature. Following incubation, the wells were washed 5 times with deionized water (provided with the kit) and kept dry. Then 100 ml substrate solution was pipetted into the antibody coated wells and incubated at room temperature for another 5 minutes. Then, 100 ml stop solution (provided with the kit) was added into antibody coated wells which changed the color of solution from blue to yellow. Finally, photometric measurement of aflatoxin was done by putting the antibody coated wells in ELISA aflatoxin reader (Chromate Plate Reader-4300/USA) at 405nm, 450 nm and 630nm. All data were calculated by using the SPSS version 9.5 (SPSS, Cary, NC, USA) statistical analysis program.

Source of Information: Bangladesh J Microbiol, Volume 35, Number 1, June 2018, pp 75-78

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

As total eradication of toxigenic fungi and their toxin is not possible, some selective approaches can be a help in aûatoxin management, like, resistant varieties development, biological control of these fungi and aflatoxin, integrated agronomy practices throughout the whole process of grain harvest, shipping, storage, feed manufacturing, and its formulation. This whole study emphasizes the need of proper surveillance and constant monitoring programs for aflatoxin free food and feedstuffs for human and animal.

Knowledge Management: Journal