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Kim, So, Song, Lee, Lim, Lee, So, Kim, Park, and Park: Comparison of Biological and Genetic Characteristics Between Sucrose-Fermenting and Sucrose-Nonfermenting Vibrio vulnificus Isolates
Original Article

J Bacteriol Virol 2006;36(4):221-228.

Published online: 18 February 2006

DOI: https://doi.org/10.4167/jbv.2006.36.4.221

Comparison of Biological and Genetic Characteristics Between Sucrose-Fermenting and Sucrose-Nonfermenting Vibrio vulnificus Isolates

Shin-Moo Kim1, Hyang-Ah So2, Kye-Min Song3, Young-Youp Lee4, Chae-Won Lim1, Jae-Hyung Lee5, Hong-Seob So5, Jin-Kyung Kim5, Rae-kil Park5, Seok-Don Park6

1Department of Clinical Laboratory Science, Wonkwang Health Science College, Iksan 570-750, Korea

2Department of Microbiology, Jellabukdo Instituite of Health & Environmental, Jeonju 561-844, Korea

3Department of Biology, Graduate School, Sunchon National University, Sunchon 540-742, Korea

4Department of Biology, Graduate School, Jeonju University, Jeonju 560-759, Korea

5Vestibulocochlear Research Center & Department of Miocrobiology, Iksan 570-749, Korea

6Dermatology, Wonkwang University School of Medicine, Iksan 570-749, Korea

Received 28 October 2006       Accepted 4 December 2006

Copyright © 2006 Journal of Bacteriology and Virology

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Twelve strains of V. vulnificus isolated from clinical specimens in 2002∼2004 in Jeollado province were determined for their biologic groups, serotypes, presence of vvhA (hemolysin/cytolysin) gene, DNA sequence, and PFGE patterns of NotI-restricted genomic DNA. The following results were obtained. All 12 isolates were biogroup 1, and API 20E profiles were: 5146105 for 5 (41.7%) isolates, and 5148125 for 2 isolates with sucrose fermentation. Ten (83.3%) of the 12 isolates was V. vulnificus serotype O4A, and two sucrose-fermenting isolates belonged to serotype O2. Alleles of cytolysin-hemolysin gene were detected in all 12 isolates. The nucleotide sequences of vvhA genes from strains WKHC 212 and WKHC 221 showed 94∼97% similarity compared with those from previously reported 7 strains, YJ016, CMCP6, L-180, CDC B3547, IF Vv10, CIP 75.4T and CNRVC 970121. PFGE of NotI-restricted genomic DNA from the 12 isolates showed approximately 48.5 to 873-kb fragments and they were clustered to five (A to E) patterns. Two sucrose-fermenting isolates belonged to pattern D with 95% similarity with each other. Two strains isolated from two different patients had two identical patterns C and D. It is concluded that sucrose-fermenting strains also exist among clinical isolates of V. vulnificus in Korea, and they can be identified by using API 20E system, and by detecting vvhA gene. DNA sequences and PFGE pattern of NotI-restricted genomic DNA suggested that the two sucrose-fermenting isolates belonged to an identical clone, and two strains each isolated from two different patients belonged to two identical clones.

Keywords: Sucrose-fermenting, Vibrio vulnifcius, Biological, Genetic characteristic

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Figure 1.
Aligned nucleotide sequences of vvhA (hemolysin/cytolysin) genes from V. vulnificus VV-212, VV-221 and previously reported 7 strains (V. vulnificus YJ016, CMCP6, L-180, CDC B3547, IF Vv10, CIP 75.4T and CNRVC 970121).
a Number refer to position from the start codon in vvhA.
jbv-36-221f1.tif
Figure 2.
Dendrogram generated with Fingerprinting II informatix software showing the PFGE types (A∼E) of NotI-digested chromosome DNA of 12 V. vulnificus isolates.
jbv-36-221f2.tif
Table 1.
Biochemical characteristics of the three biogroup strains of V. vulnificusa
Test Biogroupb
1 2 3 (Israeli vibrio)
Oxidase + + +
Arginine dihydrolase – – –
Lysine decarboxylase + + +
Ornithine decarboxylase 55 – +
Indole + – +
Citrate (Simmons) (+) + –
ONPG (+) + –
Cellobiose + + –
D-mannitol 45 – –
Lactose fermentation (+) + –
Salicin fermentation + + –
D-sorbitol fermentation – + –

a Adapted from Bisharat et al. (1999)

b Symbols: +, most (90% or greater) strains positive; (+), many strains (75 to 89.9%) positive; -, most strains negative (10% or less are positive), Numbers give the actual percentage of strains that are positive.

c ONPG, o-nitrophenyl-β-D-galactopyranoside.

Table 2.
Serotypes, biochemical characteristics and cytolysin gene of the V. vulnificus isolates
Strain no. Source Isolation area and year Serotype API20 E profile Acid from vvhA gene
Cellobiose Sucrose
VV-198 Unknown Chonnam, 2002 O4A 5146105 + – +
VV-205 Unknown Chonnam, 2002 04A 5346105 + – +
VV-208 Wound pus Chonbuk, 2003 04A 5146105 + – +
VV-209 Wound pus Chonbuk, 2003 04A 5346045 + – +
VV-210 Blood Chonbuk, 2003 04A 5146105 + – +
VV-211 Blood Chonbuk, 2003 04A 5146005 + – +
VV-212 Blood Chonnam, 2003 02 5146125 + + +
VV-221 Blood Chonnam, 2004 02 5146125 + + +
VV-CN1 Unknown Chonnam, 2003 04A 5146105 + – +
VV-CN2 Unknown Chonnam, 2003 04A 5146105 + – +
VV-CN3 Unknown Chonnam, 2003 04A 5146106 + – +
VV-CN4 Unknown Chonnam, 2003 04A 5146005 + – +
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