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Phenotypic and genetic characterization of Pasteurella multocida and related isolates from rabbits in Switzerland

Anina B.J. Stahel, Richard K. Hoop, Peter Kuhnert and Boena M. Korczak¹

Correspondence: ¹Corresponding Author: Boena M. Korczak, Institute of Veterinary Bacteriology, University of Bern, Laengggassstrasse 122, PO Box, CH-3001 Bern, Switzerland. bozena.korczak{at}vbi.unibe.ch

Several bacteria belonging to the family Pasteurellaceae are potential pathogens in rabbits. In particular, Pasteurella multocida is considered to be important, and outbreaks caused by this species result in considerable economic losses in rabbitries. However, Pasteurellaceae spp. isolated from rabbits are poorly characterized, and thus, proper identification of P. multocida isolates from these animals is problematic and often unsatisfactory, thereby hampering epidemiological investigations. Therefore, 228 isolates from rabbit populations originating from a breeding and fattening organization with group management and postmortem cases with pasteurellosis from individual owners were phenotypically and genotypically analyzed using biochemical tests and repetitive extragenic palindromic polymerase chain reaction (REP-PCR). Furthermore, 41 samples representing observed phenotypes were selected for phylogenetic analysis using 16S ribosomal RNA and rpoB genes. The REP-PCR typing and phylogenetic analyses correlated well and appeared to be distinct molecular methods for characterization of rabbit isolates. Phenotyping, however, diverged from molecular recognition, reflecting the problematic conventional diagnosis of these strains. The fermentation of sorbitol appeared to be an imprecise indicator for P. multocida subspecies classification. According to REP-PCR and sequencing results, 82% of the isolates were characterized as P. multocida subsp. multocida, 3% as P. multocida subsp. septica, and 5% as P. multocida. Further, 5% were identified as Pasteurella canis. The other 5% represented a homogeneous group of unknown species belonging to the Pasteurellaceae. Samples obtained from individual postmortem cases demonstrated a higher phenotypic and genetic heterogeneity than samples from group management rabbits.

Key Words: β subunit of RNA polymerase sequence analysis • phenotype • rabbit pasteurellosis • repetitive extragenic palindromic polymerase chain reaction • 16S ribosomal RNA gene sequence analysis