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Int J Syst Bacteriol 48 (1998), 851-858; DOI 10.1099/00207713-48-3-851
© 1998 Society for General Microbiology
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Leptospira fainei sp. nov., isolated from pigs in Australia

P. Perolat1, R. J. Chappel2, B. Adler3, G. Baranton4, D. M. Bulach3, M. L. Billinghurst2, M. Letocart1, F. Merien1 and M. S. Serrano3

1Leptospira Laboratory, Institut Pasteur, Nouméa, New Caledonia
2Victorian Institute of Animal Science, Attwood, Victoria, Australia
3Department of Microbiology, Monash University, Clayton, Victoria, Australia
4WHO Collaborating Centre on Epidemiology of Leptospirosis, Molecular and Medical Bacteriology Unit, Institut Pasteur, Paris, France

Author for correspondence: P. Perolat. Tel: +687 27 02 80. Fax: +687 27 33 90. e-mail: perolat_pasteur{at}canl.nc

ABSTRACT

Pathogenic leptospires can be causative agents of reproductive problems in pigs. Cultures of uteri and kidneys from two pig herds in New South Wales and Victoria (Australia) yielded five strains identified as Leptospira on morphological and cultural grounds. Phenotypic characteristics (growth at 13 and 30°C, growth in the presence of 8-azaguanine) were intermediate between those of pathogenic and saprophytic leptospires. No cross-agglutination was observed with reference antisera representing the 24 pathogenic serogroups and the main saprophytic ones. Antiserum against one of the strains did not agglutinate reference strains representative of any serogroup. This provided evidence of a new serovar, designated hurstbridge. Genomic characterization of the five strains was achieved using five molecular approaches. Mapped restriction site polymorphisms in the rrs (16S rRNA) gene were not related to those of any reference strains. Arbitrarily primed PCR fingerprints suggested clonality of the five strains. The strains all showed an identical and unique PFGE profile. PCR, using primers specific for the rrs gene of pathogenic leptospires, amplified corresponding sequences from the strains. DNA-DNA hybridization (and reciprocal experiments) using the S1 nuclease/TCA method was performed between one of the strains and the reference strains of Leptospira species. The homology ranged from 0 to 36% (the latter being with Leptospira inadai) thus satisfying the criterion of a new species, Leptospira fainei (type strain BUT 67). Phylogenetic analysis of 16S rRNA sequences showed that L. fainei and L. inadai formed a clade separate from the previously recognized ‘saprophyte’ and ‘pathogen’ clades.

Key Words: Leptospira fainei sp. nov. • pigs • phylogenetic analysis • serovar hurstbridge • molecular typing

The GenBank accession number for the 16S rRNA sequence of L. fainei is U60594.




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