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Journal of Clinical Microbiology, October 2009, p. 3218-3225, Vol. 47, No. 10
0095-1137/09/$08.00+0     doi:10.1128/JCM.01246-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Molecular Characterization of Capnocytophaga canimorsus and Other Canine Capnocytophaga spp. and Assessment by PCR of Their Frequencies in Dogs ^,

Alje P. van Dam,^1,* Angela van Weert,¹ Celine Harmanus,¹ K. Emiel Hovius,² Eric C. J. Claas,¹ and Frans A. G. Reubsaet³

Department of Medical Microbiology, Leiden University Medical Centre, P.O. Box 9600, 2300 RC Leiden, The Netherlands,¹ Companion Animal Hospital't Heike, 5508 PA Veldhoven, The Netherlands,² Bacterial Diagnostics Section, Diagnostic Laboratory for Infectious Diseases, National Institute of Public Health and the Environment, 3720 BA Bilthoven, The Netherlands³

Received 25 June 2009/ Accepted 22 July 2009

Capnocytophaga canimorsus can be a virulent pathogen, whereas C. cynodegmi is of low virulence. Heterogeneity within these species, their frequency in dogs, and pathogenicity factors are largely unknown. Strains from blood cultures from patients presumptively identified as C. canimorsus (n = 25) and as C. cynodegmi by rrs analysis (n = 4), blood cultures from dogs (n = 8), blood cultures from cats (n = 2), and cultures from swabs from dog mouths (n = 53) were analyzed. PCR-restriction fragment length polymorphism (PCR-RFLP), a species-specific PCR on rpoB, and rrs sequencing were used. All 29 strains from human blood cultures could be grouped into three PCR-RFLP types. One included the C. canimorsus type strain, and the other types were closely related. Two canine strains were C. canimorsus and grouped into the least common RLFP pattern group. Five were C. cynodegmi and clustered with the reference strain. One canine and both feline strains were distinct. Four human strains that presumptively had been identified as C. cynodegmi by RNA gene sequence analysis clustered with the C. canimorsus strains by both PCR-RFLP and the sequence-specific PCR of the rpoB gene. C. canimorsus DNA was present in 73% (range, 61 to 85%) of dogs' mouths, and C. cynodegmi DNA was present in 96% (range, 94 to 100%) of dogs' mouths. As defined by rpoB PCR-RFLP and by PCRs using specific primers, all strains from human blood were C. canimorsus. The sequencing of rrs genes suggested the presence of different gene copies in a few strains, indicating that the method is less appropriate for species identification. Both species are present in the majority of dogs. Additional Capnocytophaga species occur in dogs' and cats' mouths.

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* Corresponding author. Mailing address: Department of Medical Microbiology, OLVG, P.O. Box 95500, 1090 HM Amsterdam, The Netherlands. Phone: 31205993018. Fax: 31205993807. E-mail: a.p.vandam{at}olvg.nl

Published ahead of print on 29 July 2009.

Supplemental material for this article may be found at http://jcm.asm.org/.

Present address: Department of Medical Microbiology, Onze Lieve Vrouwe Gasthuis, Oosterpark 9, 1090 HM Amsterdam, The Netherlands, and Regional Laboratory for Public Health, Nieuwe Achtergracht 100, Amsterdam, The Netherlands.

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Journal of Clinical Microbiology, October 2009, p. 3218-3225, Vol. 47, No. 10
0095-1137/09/$08.00+0     doi:10.1128/JCM.01246-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.