Toxinotyping of necrotic enteritis-producing and commensal isolates of Clostridium perfringens from chickens fed organic diets

Abstract

The present study determined the effect of Clostridium perfringens isolates taken from necrotic enteritis (NE) outbreaks on organic farms in a NE virulence testing model. Thirteen strains were isolated in the course of the study. Six C. perfringens field isolates were taken from a naturally occurring NE outbreak on an organic farm. Polymerase chain reaction toxinotyping was used to establish C. perfringens strains, as well as to create a toxin profile. All field isolates were found to be type A and positive for alpha, beta-2 and netB toxin genes. During the NE virulence model, digesta samples were collected before oral inoculation to define the C. perfringens found as part of the natural flora. Three of the five natural flora isolates were found to be C. perfringens type E while the other two isolates were type A; only four of five isolates were positive for either netB or beta-2 toxin genes. Two isolates collected after inoculation were C. perfringens type A positive for cpb2 and netB. All isolates were tested positive for the quorum-sensing-related gene luxS, regardless of the strain source. The presence of luxS, alpha, netB and beta-2 toxin genes seems not to be a determinant of the disease as they were present in isolates from both outbreak birds as well as healthy and pre-inoculated birds. The C. perfringens field isolates induced mild NE lesions in one-half of the birds during the challenge study. Other mechanisms must play a role in the development of the disease beyond toxinotype, potentially including intestinal ecology and health, which would account for acute disease as seen in the field outbreak.

Acknowledgements

The excellent technical assistance of Dr Tom Hutchison and Chris Link-Muirhead from Manitoba Agriculture, Food and Rural Initiatives, Veterinary Diagnostic Services (MAFRI) is greatly appreciated. The authors would like to thank Andre Hamel (MAFRI) for providing the control PCR C. perfringens strains, Angela Kroeker for incredible technical support, and Omar Abu-Dahab for providing the certified organic feed. The authors would like to express appreciation to the Agricultural and Rural Development Initiative (ARDI) for the ?nancial support from a scholarship for the ?rst author. Financial support from the Manitoba Egg Farmers for poultry infrastructure resources is gratefully appreciated.