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Journal of Clinical Microbiology, December 2008, p. 3957-3964, Vol. 46, No. 12
0095-1137/08/$08.00+0     doi:10.1128/JCM.01548-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Multilocus Sequence Typing Analysis of Clostridium perfringens Isolates from Necrotic Enteritis Outbreaks in Broiler Chicken Populations{triangledown}

G. Chalmers,1 H. L. Bruce,2,{dagger} D. B. Hunter,1 V. R. Parreira,1 R. R. Kulkarni,1 Y.-F. Jiang,1 J. F. Prescott,1 and P. Boerlin1,3*

Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, Ontario N1G 2W1, Canada,1 Nutreco Canada Agresearch, 473-Sixth Concession Road, R.R. #3, Burford, Ontario N0E 1A0, Canada,2 Laboratory for Foodborne Zoonoses, Public Health Agency of Canada, Guelph, Ontario N1G 3W4, Canada3

Received 11 August 2008/ Returned for modification 13 September 2008/ Accepted 10 October 2008

Clostridium perfringens is an important pathogen of animals and humans and is the causative agent of necrotic enteritis (NE) in poultry. This study focuses on the typing of intestinal C. perfringens isolates (n = 61) from outbreaks of NE collected from several areas of Southern Ontario, using a recently developed multilocus sequence typing (MLST) technique. For comparison, C. perfringens isolates from healthy birds were also obtained and typed. An additional locus, the pfoS locus, was included in our analysis, in an attempt to increase the discriminatory ability of the method previously published. Birds were collected from two major poultry processors in Canada, and isolates from processor 2 formed a distinct MLST cluster. Isolates from healthy birds also collected from the outbreak flocks clustered together with isolates from the birds with NE. Although isolates from eight outbreaks clustered together, MLST types were also occasionally different between outbreaks. Strong linkage disequilibrium was observed between loci, suggesting a clonal C. perfringens population structure. Detection assays for toxin genes cpb2 (beta-2 toxin), tpeL, and the newly described netB (NetB toxin) were also performed. netB was almost always found in outbreak isolates, whereas cpb2 was found exclusively in healthy bird isolates. The toxin gene tpeL, which has not been previously identified in C. perfringens type A strains, was also found, but only in the presence of netB. Resistance to bacitracin was found in 34% of isolates from antimicrobial agent-free birds and in 100% of isolates from conventionally raised birds.

* Corresponding author. Mailing address: Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, Ontario N1G 2W1, Canada. Phone: (519) 824-4120, ext. 54647. Fax: (519) 824-5930. E-mail: pboerlin{at}uoguelph.ca

{triangledown} Published ahead of print on 22 October 2008.

{dagger} Present address: Department of Agriculture, Food and Nutritional Science, University of Alberta, Edmonton, Alberta T6G 2P5, Canada.

Journal of Clinical Microbiology, December 2008, p. 3957-3964, Vol. 46, No. 12
0095-1137/08/$08.00+0     doi:10.1128/JCM.01548-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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