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Applied and Environmental Microbiology, March 2009, p. 1604-1613, Vol. 75, No. 6
0099-2240/09/$08.00+0     doi:10.1128/AEM.01594-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Evaluation of Genotypic and Phenotypic Methods To Distinguish Clinical from Environmental Vibrio vulnificus Strains{triangledown} ,{dagger}

Eva Sanjuán,1,{ddagger} Belén Fouz,1 James D. Oliver,2 and Carmen Amaro1*

Departamento de Microbiología y Ecología, Universidad de Valencia, Avda. Dr. Moliner 50, 46100 Burjassot, Valencia, Spain,1 Department of Biology, University of North Carolina at Charlotte, Charlotte, North Carolina 282232

Received 11 July 2008/ Accepted 5 January 2009

Vibrio vulnificus is a heterogeneous bacterial species that comprises virulent and avirulent strains from environmental and clinical sources that have been grouped into three biotypes. To validate the typing methods proposed to distinguish clinical from environmental isolates, we performed phenotypic (API 20E, API 20NE, and BIOLOG tests) and genetic (ribotyping and DNA polymorphism at several loci) studies with a large strain collection representing different biotypes, origins, and host ranges. No phenotypic method was useful for biotyping or grouping strains with regard to the origin of an isolate, and only the BIOLOG system was reliable for identifying the strains at the species level. DNA polymorphisms divided the population into three major profiles. Profile 1 strains were vcg type C, 16S rRNA type B, and vvh type 1 and included most of the biotype 1 human septicemic isolates; profile 2 strains were vcg type E, 16S rRNA type A, and vvh type 2 and included all biotype 2 isolates together with biotype 1 isolates from fish and water and some human isolates; and profile 3 strains were vcg type E, 16S rRNA type AB, and vvh type 2 and included biotype 3 strains. Ribotyping divided the species into two groups: one group that included profile 1 biotype 1 isolates and one group that included isolates of all three biotypes with the three profiles described above. In conclusion, no genotyping system was able to distinguish either clinical strains from environmental strains or biogroups within the species V. vulnificus, which suggests that new typing methodologies useful for public health have to be developed for this species.

* Corresponding author. Mailing address: Departamento de Microbiología y Ecología, Universidad de Valencia, Avda. Dr. Moliner 50, 46100 Burjassot, Valencia, Spain. Phone: 34-96-3543104. Fax: 34-96-3544570. E-mail: carmen.amaro{at}uv.es

{triangledown} Published ahead of print on 9 January 2009.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

{ddagger} Present address: South Texas Center for Emerging Infectious Diseases and Department of Biology, The University of Texas at San Antonio, One UTSA Circle, San Antonio, TX 78249.

Applied and Environmental Microbiology, March 2009, p. 1604-1613, Vol. 75, No. 6
0099-2240/09/$08.00+0     doi:10.1128/AEM.01594-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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