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
