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Detection of Aflatoxigenic Fungi and Aflatoxin Levels in Poultry Feed

Marufa Khatun
Department of Microbiology, Faculty of Veterinary and Animal Science, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh.

Farzana Afroz
Department of Microbiology, Faculty of Veterinary and Animal Science, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh.

Md. Khaled Hossain
Department of Microbiology, Faculty of Veterinary and Animal Science, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh.

Abstract/ Executive Summary:

The present research work was carried out for the detection of aflatoxigenic fungus from poultry feed samples. During a period of 12 months (from June 2018 to June 2019), a total of 100 poultry feed samples comprising of 50 commercially prepared and 50 self compounded poultry feed were collected from different 5 different farms of Dinajpur districts, Bangladesh. Among the 100 feed samples, Aspergillus spp was found on 54 feed samples with 54% prevalence. But Aflatoxigenic Aspergillus spp was found on 48 samples with 48% prevalence. In this study, the prevalence of Aspergillus spp was 56% in self compounded poultry feed and 52% prevalence in commercially prepared poultry feed. There was no significant difference in the prevalence of Aspergillus spp and also in the prevalence of aflatoxigenic fungus isolated from the commercially prepared feed as well as in self compounded feed on the basis of farms. The maximum level of aflatoxin was ≥20 ppb. This study alarms us about the potential risks of Aspergillus spp to public health if contaminates agricultural commodities such as grains or raw materials such as poultry feed.

Keywords: Aspergillus spp, Aflatoxin, Poultry Feed, Potential risks

Location: Dinajpur district, Bangladesh.

Start Date: 00-06-2018

End Date: 00-06-2019

Program Area: Animal Health and Management

Commodity: Poultry, Fungus, Feed

Objectives:

The present study was undertaken for the isolation and identification of aflatoxin-producing fungus and also for the determination of aflatoxin levels in poultry feed samples in Bangladesh.

Methodology:

A total of 100 samples (50 commercially prepared feed samples and 50 self-compounded feed samples) were collected from different local farms of Dinajpur district, Bangladesh, and brought to the microbiology laboratory, HSTU. Approximately 300 g of each feed sample were collected and kept in an ice-box during transportation to the laboratory and stored at 4°C until testing. They were analyzed within 24 hours of sampling. 300 gm of different poultry feed samples were uniformly homogenized in mortar and pastle using a sterile diluent as per the recommendation of ISO, 1995. A homogenized suspension was made with the help of mortar and pastle. A quantity of 10 ml homogenate samples were transferred carefully into a sterile test tube containing 90 ml of PBS. Thus 1:10 dilution of the samples were obtained. With the help of a sterile inoculating loop the processed samples were inoculated into the Sabroud Dextrose Agar (SDA) and incubated at 37°C for 5-7 days. Colonies from Sabroud Dextrose Agar were subcultured in Potato Dextrose Agar Media. Suspected colonies stained by lactophenol cotton blue (LPCB) were examined microscopically for identification of Aspergillus spp. Micromorphological characteristics were observed by wet mount in lactophenol cotton blue stain for identification by the conidiospore appearance and arrangement. Aspergillus Differential Agar Base (ADAB) was used to detect aflatoxin producing ability of the fungal isolates. Observed specific colonies from Sabroud Dextrose Agar and Potato Dextrose agar were subcultured in Aspergillus flavus parasiticus agar medium. The bright orange colour on the reverse side of the plates of Aspergillus Differentiation Medium Base will indicate a positive result. In this study aflatoxigenic fungus was detected by Agra Strip total aflatoxin test 20ppb cut-off. The Agrastrip total Aflatoxin test is a one-step lateral flow immune-chromatographic assay that determines a qualitative level for the presence of total aflatoxin in grains, cereals, feeds and other commodities. It detects the presence of aflatoxin at 5 ppb or higher in grain samples by utilizing highly specific reactions between antibodies and aflatoxin in grain samples. Antibody-particle complex is dissolved in assay diluent and mixed with sample extract. The mixed content is then wicked onto a membrane, which contains a test zone and a control zone. The test zone captures a free antibody-particle complex, allowing color particles to concentrate and form a visible line. A positive sample with aflatoxin above the cutoff level will result in no visual line in the test zone. Alternatively, a negative sample with aflatoxin below the cutoff level will form a visible line in the test zone. The line will always be visible in the control zone regardless of the presence of aflatoxin. A representative sample was grinded using a Romer Series II® Mill so that 75% will pass through a 20-mesh screen, then thoroughly mix the subsample portion.10 g of ground sample were taken into a clean jar with 20 mL of 50% ethanol extraction solution (i.e. 50/50 (v/v) ethanol/water) and the jar was sealed and vortexed for 1 minute. The top layer of extract through which the sample was filtered by the Whatman filter was collected. All reagents and kit components must be at room temperature 18-30°C (64-86°F) before use. Using a single channel pipette, 50μL of assay diluent to each microwell was added. The coating conjugate was dissolved in the microwell by pipetting the content up and down 5 times. 50μL of sample extracts was added to each microwell, mixing the content in each well by pipetting it up and down 3 times. One test strip was put into one well. The test strip was allowed to develop color for 5 minutes. Then the result was interpreted according to the manufacturer.

Source of Information: International Journal of Natural and Social Sciences, 2021, 8(2): 74-80

Article Link (Online Journal): DOI: 10.5281/zenodo.5168502

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Aflatoxin contamination in feed is a serious risk for public health having long-term health effects. Most of the crops used in culinary in Bangladesh have been reported to be contaminated with aflatoxigenic Aspergillus flavus and/or aflatoxins. Continuous surveillance should be conducted to detect aflatoxin-contaminated crops and contamination levels in different poultry feeds. Good agricultural management practice should be employed to reduce the contamination risk of aflatoxins and aflatoxigenic fungi in poultry feed. As total eradication of toxigenic fungi and their toxin is not possible, some selective approaches can be a help in aflatoxin management, like, resistant varieties development, biological control of these fungi and aflatoxin, integrated agronomy practices throughout the whole process of the grain harvest, shipping, storage, feed manufacturing, and its formulation. This whole study emphasizes the need for proper surveillance and constant monitoring programs for aflatoxin-free food and feedstuffs for humans and animals.

Knowledge Management: Journal