Clonal stability of Pasteurella multocida in free-range layers affected by fowl cholera

Detailed longitudinal studies of the genetic stability of Pasteurella multocida ssp. multocida, the cause of fowl cholera, have not previously been carried out. Consequently, the aim of the present study was to provide detailed information on the genetic stability and diversity of P. multocida ssp. multocida in poultry flocks over time, enabling new insights into the molecular epidemiology of this important poultry pathogen. Longitudinal investigations of the rate and causes of mortality were carried out on two free-range layer farms (A and B) over a period of 11 months. The total mortality of two flocks, A1 and A2, on farm A were 62 and 91%, respectively, while the total mortality of a single flock B1 on farm B was 6%. Postmortem examinations were performed on 708 layers from flocks A1 and A2 and in 159 from flock B1. Fowl cholera was the main cause of mortality on both farms. Pasteurella multocida isolates recovered from layers on both farms were characterized phenotypically and genotypically, and 322 isolates were identified as P. multocida ssp. multocida. The genetic diversity of 99 isolates from farm A and 31 from farm B was characterized by restriction endonuclease analysis and amplified fragment length polymorphism analysis. The isolates on each farm had a unique restriction endonuclease analysis and amplified fragment length polymorphism analysis type, suggesting a single introduction of a successful clone. Furthermore the clone on farm A was identical to clones previously isolated from outbreaks in the avifauna of Denmark in 1996 and 2001 and in Sweden in 1998. This study provides convincing evidence for the clonal stability of outbreak clones of P. multocida.