Detection of alpha- 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 enterotoxaemia 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 with pathological signs of enterotoxaemia showed the presence of the alpha- and epsilon-toxin genes but were devoid of the beta-toxin gene. These strains could therefore be identified as type D, characteristic for clostridial enterotoxaemia of sheep, lambs and goats. In contrast, Cl. perfringens isolated from 11 of 13 healthy animals only contained the alpha-toxin gene which is typical for type A. Two of the healthy animals contained Cl. perfringens with the alpha- and epsilon-toxin genes. However, when several individual Cl. perfringens colonies were analysed from each of these two animals, only a small percentage was found to contain the epsilon-toxin gene, whereas the majority of the colonies were of type A with the alpha-toxin gene only. This is in contrast to the findings from the diseased animals which contained practically only type D Cl. perfringens. The beta-toxin gene was not found in any Cl. perfringens isolate from goat and sheep. Comparison of the PCR data with results obtained by the classical biological toxin assay using the mouse model showed a good correlation.
Institute for Veterinary Bacteriology, University of Berne, Switzerland.
R
R
Miserez
J
J
Frey
C
C
Buogo
S
S
Capaul
A
A
Tontis
A
A
Burnens
J
J
Nicolet
D000818:Animals; D001427:Bacterial Toxins / Q000032:analysis / Q000235:genetics; D002135:Calcium-Binding Proteins; D003016:Clostridium perfringens / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000032:analysis; D004767:Enterotoxemia / Q000382:microbiology*; D015511:Goat Diseases / Q000382:microbiology*; D006041:Goats; D007422:Intestines / Q000382:microbiology; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods*; D012756:Sheep; D012757:Sheep Diseases / Q000382:microbiology*; D010738:Type C Phospholipases / Q000032:analysis / Q000235:genetics
D001427:Bacterial Toxins; D002135:Calcium-Binding Proteins; C039811:Clostridium perfringens epsilon-toxin; D004269:DNA, Bacterial; D010738:Type C Phospholipases; C040649:alpha toxin, Clostridium perfringens