DETECTION OF ALPHA-TOXIGENIC AND EPSILON-TOXIGENIC CLOSTRIDIUM-PERFRINGENS TYPE-D IN SHEEP AND GOATS USING A DNA AMPLIFICATION TECHNIQUE (PCR)

Clostridium perfringens isolated from sheep and goat with enterotoxaem ia at necropsy and from healthy animals at slaughter were typed using specific PCR assays for the detection of the alpha-, beta- and epsilon -toxin genes. Clostridium perfringens isolated from all 52 animals wit h pathological signs of enterotoxaemia showed the presence of the alph a- and epsilon-toxin genes but were devoid of the beta-toxin gene. The se strains could therefore be identified as type D, characteristic for clostridial enterotoxaemia of sheep, lambs and gears. In contrast, CI . perfringens isolated from 11 of 13 healthy animals only contained th e alpha-toxin gene which is typical for type A. Two of the healthy ani mals contained Cl. perfringens with the alpha- and epsilon-toxin genes . However, when several individual CI. perfringens colonies were analy sed from each of these two animals, only a small percentage was found to contain the epsilon-toxin gene, whereas the majority of the colonie s were of type A with the alpha-toxin gene only. This is in contrast t o the findings from the diseased animals which contained practically o nly type D CI. perfringens. The beta-toxin gene was not found in any C l. perfringens isolate from goat and sheep. Comparison of the PCR data with results obtained by the classical biological toxin assay using t he mouse model showed a good correlation.