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Identification of Vibrio cholerae and Vibrio mimicus by multilocus sequence analysis (MLSA)

¹ Instituto Oswaldo Cruz, Rio de Janeiro, Brazil
² Department of Genetics, Federal University of Rio de Janeiro, Brazil

Correspondence
Cristiane C. Thompson
thompson{at}ioc.fiocruz.br

Vibrio cholerae and Vibrio mimicus have similar phenotypes and genomes making rapid differentiation of these two species difficult. The first standard multilocus sequence analysis (MLSA) scheme for the identification of these species is described. A collection of 45 representative isolates from different geographical regions and hosts was examined using segments of the housekeeping genes pyrH, recA and rpoA. Overall, the closest phylogenetic neighbours of these species were Vibrio furnissii and Vibrio fluvialis. V. cholerae and V. mimicus formed separate species clusters on the basis of each gene, suggesting that these genes are useful as identification markers. These species clusters arose by the accumulation of point mutations. The pyrH gene showed the highest resolution for differentiating V. cholerae and V. mimicus. The maximum interspecies pyrH gene sequence similarity was 91 %. Clearly, V. mimicus strains were more heterogeneous than V. cholerae strains at the three loci. It is suggested that vibrio species may be defined on the basis of MLSA data. A vibrio species was defined as a group of strains forming a monophyletic group on the basis of these loci and with an intraspecific sequence similarity of at least 95 %. V. cholerae and V. mimicus isolates can be readily identified through the open database resource ‘The Taxonomy of Vibrios’ (http://www.taxvibrio.lncc.br/).

The GenBank/EMBL/DDBJ accession numbers for the pyrH, recA and rpoA gene sequences of Vibrio cholerae and Vibrio mimicus are EF990257–EF990363 and EU147787–EU147789.

Tables detailing the sources of the strains used in this study, the primers used for amplification and sequencing of the three loci, features of the genes and signatures of the V. cholerae and V. mimicus pyrH, recA and rpoA genes are available with the online version of this paper. Information is also available on how the recombination analysis was performed.