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Serological and molecular characteristics of Vibrio vulnificus biotype 3: evidence for high clonality

Naiel Bisharat^1,

¹ Department of Epidemiology and Preventive Medicine, School of Public Health, Sackler Faculty of Medicine, Tel Aviv University, Ramat Aviv, Israel
² Departamento de Microbiologia y Ecologia, Campus de Burjassot, Universidad de Valencia, Valencia 46100, Spain

Correspondence
Naiel Bisharat
bisharat_na{at}clalit.org.il

Vibrio vulnificus biotype 3 has been implicated as the causative pathogen of an ongoing disease outbreak that erupted in Israel in 1996. Recent work based on multi-locus sequence typing (MLST) showed that V. vulnificus biotype 3 is genetically homogeneous. The aim of this study was to investigate the existence of subpopulations within this homogeneous biotype by characterizing the surface antigens and analysing the sequence diversity of selected outer-membrane protein (OMP)-encoding genes. Rabbit antisera were prepared against biotype 1, 2 and 3 strains. The results of the slide-agglutination test, dot-blot assay (using fresh and boiled cells), and immunoblotting of lipopolysaccharides (LPS) and OMPs were evaluated. By slide-agglutination and dot-blot assays all biotype 3 strains agglutinated with the selected biotype 3 strain. This homogeneity was supported by immunoblot analysis of the LPS. Analysis of OMP patterns revealed that all three biotypes share a considerable number of common bands that are antigenically related. Cluster analysis of DNA sequence data from selected OMP-encoding genes showed that biotype 3 strains form a genetically distinct and homogeneous clone. The homogeneity of surface antigens and the lack of any sequence diversity among both housekeeping and OMP-encoding genes reaffirms the highly clonal nature of biotype 3 and suggests that it has only recently descended from the parent population of V. vulnificus.

Part of this work was done at the Nuffield Department of Clinical Laboratory Sciences, University of Oxford, Oxford, UK.

The GenBank/EMBL/DDBJ accession numbers for the vuuA sequences reported in this paper are DQ973529, DQ973530 and DQ980539; for the wcvI sequences, DQ980540 and DQ980541; for the wza sequences, DQ980542 and DQ980543; and for the ompU sequences, DQ980544, DQ980545 and DQ980546.

Present address: Department of Medicine, section D, Ha'Emek Medical Center, Afula 18101, Israel.

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