Genotyping of Clostridium perfringens isolated from calves with neonatal diarrhea

• Published in: Anaerobe Vol. 14; no. 6; pp. 328 - 331
• Main Authors: Ferrarezi, Marina C., Cardoso, Tereza C., Dutra, Iveraldo S.
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01-12-2008
• Subjects: Animals; Animals, Newborn; Bacterial Toxins - genetics; Calves; Cattle; Cattle Diseases - microbiology; Clostridium perfringens; Clostridium perfringens - classification; Clostridium perfringens - genetics; Clostridium perfringens - isolation & purification; Diarrhea - microbiology; Diarrhea - veterinary; DNA, Bacterial - genetics; Female; Genotype; Male; Neonatal diarrhea; PCR; Polymerase Chain Reaction - methods; Calves; Neonatal diarrhea; Clostridium perfringens; PCR
• ISSN: 1075-9964; 1095-8274
• DOI: 10.1016/j.anaerobe.2008.12.001

Neonatal diarrhea is one of the main causes of losses in cattle herds.
Clostridium perfringens is a widespread enteropathogen, and is responsible for many animal diseases such as bovine neonatal diarrhea. Fecal samples from 141 diarrheic calves and 129 healthy calves, aged up to 28 days and belonging to three herds were examined. Rates of culture positivity were 36.2% and 30.2% for diarrheic and nondiarrheic calves, respectively. Multiple isolates from primary isolation plates were subjected to simultaneous genotyping by multiplex PCR, with primers amplifying fragments of alpha (
cpa), beta (
cpb), epsilon (
etx), iota (
itxA), enterotoxin (
cpe) and beta2 (
cpb2) toxin-encoding genes. Only 17/51 (33.3%) and 17/39 (43.6%) of these mixtures from diarrheic and nondiarrheic calves, respectively, yielded genotype information, suggesting that this may not be a viable approach to genotyping of isolates. Fourteen isolate mixtures from animals with diarrhea had only
cpa (type A), one had
cpa and
cpb2 (type A beta2 positive), one with
cpa,
itxA, and
cpb2 (type E, beta2 positive), and one with
cpa,
etx,
itxA, and
cpb2 toxin producing strains. Among 17 isolate mixtures from healthy calves, 10 were exclusively type A, one was type A
cpb2 positive, two were type E, three were type E
cpb2 positive, and one was types D and E
cpb2 positive. There was no correlation between isolation of a given toxin type and the presence of diarrhea.