Veterinary World


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Copyright: The authors. This article is an open access article licensed under the terms of the Creative Commons Attribution License ( which permits unrestricted use, distribution and reproduction in any medium, provided the work is properly cited.

Original Research

10. Alpha toxin specific PCR for detection of toxigenic strains of Clostridium perfringens in poultry - Malmarugan Shanmugasamy and Johnson Rajeswar
Vet World. 2012; 5(6): 365-368

Aim : Isolation of Clostridium perfringens from necrotic enteritis cases in poultry and confirmation by alpha toxin specific PCR.

Materials and Methods: Robertson cooked meat medium with Brain Heart Infusion broth was used for isolation of C. perfringens from intestinal contents of necrotic enteritis suspected birds. Positive cultures from perfringens agar were further confirmed by biochemical tests and subjected to alpha toxin specific PCR. Results: Twenty Clostridium perfringens isolates were isolated from intestinal contents of thirty five NE suspected birds. Out of the twenty isolates, fourteen were isolated from commercial broilers of 2 to 6 wk of age and six from commercial layers of 9 to 15 wk of age. Frequency of isolation of C. perfringens was more with Robertson cooked meat medium with BHI broth than thioglycollate broth alone. When positive cultures were streaked on to clostridial agar appreciable luxuriant growths were obtained and the selective streaking of these colonies on perfringens agar with supplements revealed rough and black colonies with sulphate reduction. The isolates produced rough and black colonies with sulphate reduction on perfringens agar, double zone haemolysis on sheep blood agar, stormy clot fermentation on milk medium and opalescence on egg yolk medium. The isolates were found negative for oxidase, catalase, liquefied gelatin, fermented glucose, maltose, lactose and sucrose except mannitol. All the fourteen isolates obtained from commercial broilers proved the alpha toxin producing strains of C. perfringens when they were subjected to alpha toxin specific PCR. Conclusion: This study revealed alpha toxin specific PCR is highly useful for detection of toxigenic strains of Clostridium perfringens in poultry.

Keywords: Alpha toxin, Clostridium perfringens, Necrotic enteritis, PCR, Poultry