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Applied and Environmental Microbiology, June 2003, p. 3203-3212, Vol. 69, No. 6
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.6.3203-3212.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Population Genetics of Vibrio vulnificus: Identification of Two Divisions and a Distinct Eel-Pathogenic Clone

Michaela Gutacker,^1, Nadine Conza,¹ Cinzia Benagli,¹ Ambra Pedroli,¹ Marco Valerio Bernasconi,^1, Lise Permin,^2, Rosa Aznar,³ and Jean-Claude Piffaretti^1*

Istituto Cantonale di Microbiologia, 6501 Bellinzona, Switzerland,¹ The Royal Veterinary and Agriculture University, Frederiksberg, Denmark,² Departamento de Microbiología, Facultad de Biología, Universitad de Valencia, Valencia 46100, Spain³

Received 10 October 2002/ Accepted 26 February 2003

Genetic relationships among 62 Vibrio vulnificus strains of different geographical and host origins were analyzed by multilocus enzyme electrophoresis (MLEE), random amplification of polymorphic DNA (RAPD), and sequence analyses of the recA and glnA genes. Out of 15 genetic loci analyzed by MLEE, 11 were polymorphic. Cluster analysis identified 43 distinct electrophoretic types (ETs) separating the V. vulnificus population into two divisions (divisions I and II). One ET (ET 35) included all indole-negative isolates from diseased eels worldwide (biotype 2). A second ET (ET 2) marked all of the strains from Israel isolated from patients who handled St. Peter's fish (biotype 3). RAPD analysis of the 62 V. vulnificus isolates identified 26 different profiles separated into two divisions as well. In general, this subdivision was comparable (but not identical) to that observed by MLEE. Phylogenetic analysis of 543 bp of the recA gene and of 402 bp of the glnA gene also separated the V. vulnificus population into two major divisions in a manner similar to that by MLEE and RAPD. Sequence data again indicated the overall subdivision of the V. vulnificus population into different biotypes. In particular, indole-negative eel-pathogenic isolates (biotype 2) on one hand and the Israeli isolates (biotype 3) on the other tended to cluster together in both gene trees. None of the methods showed an association between distinct clones and human clinical manifestations. Furthermore, except for the Israeli strains, only minor clusters comprising geographically related isolates were observed. In conclusion, all three approaches (MLEE, RAPD, and DNA sequencing) generated comparable but not always equivalent results. The significance of the two divisions (divisions I and II) still remains to be clarified, and a reevaluation of the definition of the biotypes is also needed.

Present address: Laboratory of Human Bacterial Pathogenesis, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT 59840.

Present address: Zoologisches Museum der Universität, 8057 Zurich, Switzerland.

Present address: DAKO A/S, Glostrup, Denmark.

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