Differentiation of Campylobacter species by AFLP fingerprinting

Institute for Animal Science and Health (ID-Lelystad), PO Box 65, 8200 AB Lelystad, The Netherlands 1 Laboratorium voor Microbiologie, University of Gent, Gent, Belgium 2 Author for correspondence: Birgitta Duim. Tel: +31 320 238157. Fax: +31 320 238153. e-mail: b.duim{at}id.wag-ur.nl The fluorescent amplified fragment length polymorphism (AFLP) fingerprinting method was tested for its ability to identify and subtype the most important Campylobacter species found in veterinary infections. Sixty-nine reference strains and 19 clinical isolates of Campylobacter jejuni subsp. jejuni , Campylobacter jejuni subsp. doylei , Campylobacter upsaliensis , Campylobacter coli , Campylobacter lari , Campylobacter fetus subsp. fetus , C. fetus subsp. venerealis , Campylobacter hyointestinalis subsp. hyointestinalis , C. hyointestinalis subsp. lawsonii , Campylobacter mucosalis , Campylobacter helveticus and Campylobacter sputorum were subjected to analysis. The topology of the dendrogram obtained by numerical analysis of the AFLP profiles did not reflect the phylogenetic relationships as derived from 16S rDNA sequence comparison. However, except for C. lari , AFLP analysis grouped the strains that belonged to the same genomic species into distinct clusters. C. lari strains were separated into two distinct AFLP groups, which corresponded with nalidixic-acid-sensitive and -resistant variants of C. lari . These results correlated with data from whole-cell protein profiling. Within C. jejuni , C. hyointestinalis and C. fetus , strains could be identified at the subspecies level. AFLP analysis also allowed the subtyping of most species at the strain level. It is concluded that AFLP analysis is a valuable tool for concurrent identification of campylobacters at the species, subspecies and strain levels. In addition, the data confirm and extend previous reports showing that C. lari is a heterogeneous species that may comprise multiple taxa. Keywords: taxonomy, typing, epidemiology, veterinary infections Abbreviations: AFLP, amplified fragment length polymorphism; UPGMA, unweighted pair-group method using arithmetic averages