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Lethal Effects of Salmonella Toxin Isolated from Layer Chickens

Farzana Afroz
Department of Microbiology, Hajee Mohammed Danesh Science and Technology University, Dinajpur-5200, Bangladesh.

Md. Mostafizer Rahman
Department of Microbiology, Hajee Mohammed Danesh Science and Technology University, Dinajpur-5200, Bangladesh.

Md. Fakhruzzaman*
Department of Microbiology, Hajee Mohammed Danesh Science and Technology University, Dinajpur-5200, Bangladesh.

Mezbah Uddin
Department of Microbiology and Hygiene, Sylhet Agricultural University, Sylhet, Bangladesh.

Abstract/ Executive Summary:

This study was conducted to determine lethal effects of Salmonella toxin isolated from layer chickens. The chickens were collected from Paragon Poultry Farm, Rangpur and Ahana Poultry Farm, Dinajpur and brought to the Microbiology Laboratory at the Hajee Mohammad Danesh Science and Technology University, Dinajpur, to isolate and identify Salmonella spp., and subsequent toxin separation from the isolated organisms for the detection of lethal effects of the toxin. The lethality of isolated toxin was evaluated in day-old-chick. The toxicity in case of oral administration was 50%, whereas the rate was 80% when chicks were administered through intra-peritoneal route. The chick mortality rate was 65%. For toxin extraction, Salmonellae were isolated and identified from layer chickens. Out of 196 organ samples, 51 were found to be positive for Salmonella spp., of which 33 and 18 were of Paragon Poultry Farm and Ahana Poultry Farm, respectively. Total 60 chickens (27 pullets and 33 layers) were screened. The percentages of positive samples (liver, spleen, lung, heart) from these farms were 29.82 and 20.73%, respectively and the average prevalence of Salmonellae was 26.02%. In case of Paragon Poultry Farm, the prevalence of salmonellosis in pullet was 23.08% and in layer, it was 35.48%, whereas in case of Ahana Poultry Farm, the prevalence was 16.67 and 23.91%, respectively.

Keywords: Salmonella toxin, Lethal effect, Layer chickens

Location: Paragon Poultry Farm (layer), Rangpur and at Ahana Poultry Farm (layer), Dinajpur.

Start Date: 00-03-2011

End Date: 00-08-2011

Program Area: Animal Health and Management

Commodity: Chicken

Objectives:

The present study was undertaken to detect the lethal effects of toxin extracted from Salmonellae field isolates that could be a candidate in future to use as toxoid vaccine.

Methodology:

Study area and duration The study was conducted during the period from March, 2011 to Aug, 2011 at Paragon Poultry Farm (layer), Rangpur and at Ahana Poultry Farm (layer), Dinajpur. The samples were collected from birds of selected layer farms and brought to the Microbiology Laboratory of the Department of Microbiology, Hajee Mohammad Danesh Science and Technology University (HSTU), Dinajpur, for laboratory analysis. Isolation and identification of Salmonellae from chicken Cultural characterization: A total of 196 field samples comprising liver, spleen, heart and lung of 60 dead chickens of study areas were aseptically collected and brought to the Microbiology Laboratory of the Department of Microbiology, HSTU. Isolation and identification of Salmonellae were performed as per procedures described by OIE (2000), Merchant and Packer (1967) and Cowan (1985). Primary culture was performed in Nutrient agar and Nutrient broth media. Sub-culturing was performed in eosin methylene blue (EMB) agar, MacConkey (MC) agar, salmonella-shigella (SS) agar and brilliant green (BG) agar to obtain pure culture and to study the cultural characteristics. Morphological characterization: The representative Salmonellae isolates from SS agar were stained by Gram staining (Merchant and Packer, 1967). Motility test was performed by Hanging drop preparation. Biochemical characterization: Isolated organisms with supporting growth characteristics of Salmonellae on various culture media were maintained on SS agar and BG agar and were subjected to biochemical tests viz. sugar fermentation test, triple sugar iron (TSI) agar slant reaction, methyl red-Voges Proskauer (MR-VP) test, indole reaction, and motility indole urease (MIU) test as described by Merchant and Packer (1967). Toxin extraction from Salmonellae field isolates: The isolated Salmonellae were inoculated in Nutrient broth culture media and kept overnight at 370C for 24 hours in bacteriological incubator. Growth of Salmonellae in broth culture was mixed with an equal volume of phosphate buffer saline (PBS) in test tubes. The test tubes were placed in a rotary shaker at room temperature for 8 hours and centrifuged at 3,000 rpm for 30 minutes. The supernatant fluids were then passed through 0.45 μm millipore filter for the separation of toxin from the bacteria. The filtrates were then mixed with saturated solution of ammonium sulphate at room temperature and centrifuged at 3,000 rpm for 30 minutes. Detection of lethal effect of Salmonellae toxin: Lethal effect of Salmonella toxin was detected in day-old chick. Thirty chickens were taken on random basis, out of them one group of ten 1-day-old SPF chicks were orally inoculated with 200 μl Salmonella toxin extracted from field isolate. Another group of 10 SPF chicks were inoculated through intra-peritoneal route with 2 ml Salmonella toxin extracted from field isolate, and 10 SPF chickens were kept as control. In case of oral administration, the dose of toxin was 200 μl but it was only 2 ml in case of intra-peritoneal administration due to variation in route. After 1 or 2 days, chicks that died were subjected to postmortem examination and organs (liver, spleen and cecal tonsils) were collected to see the postmortem changes.

Source of Information: Microbes and Health, June 2012, 1(1): 19-22 ISSN: 2226-0153 (Print)

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

In the present study, Salmonella species were successfully isolated from layer chickens. In addition, the toxin was extracted and the lethal effects of the toxin was studied using oral and intra-peritoneal route in day-old-chicks. The average prevalence of Salmonellae in the study area was 26.02%. The extracted toxin was found to be lethal for day-old-chick and the average 65% toxicity was found irrespective of route of administration. However, the toxicity was higher when the toxin was administered through intra-peritoneal route than that of oral route. Among the internal organs, Salmonellae were found to be more prevalent in liver followed by spleen, heart and lungs.

Knowledge Management: Journal