Journal List > Korean J Lab Med > v.30(5) > 1011682

TOOLS
Kim, Kim, and Shin: Virulence Characteristics of Sucrose-fermenting Vibrio vulnificus Strains
Brief Communication

Korean J Leg Med 2010;30(5):507-510.

Published online: 14 January 2010

DOI: https://doi.org/10.3343/kjlm.2010.30.5.507

Virulence Characteristics of Sucrose-fermenting Vibrio vulnificus Strains

Seong-Jung Kim, M.D.1, ∗, Choon-Mee Kim, Ph.D.2, ∗, Sung-Heui Shin, M.D.2, 3

1Department of Emergency Medicine, Gwangju, Korea

2Research Center for Resistant Cells, Gwangju, Korea

3Department of Microbiology, Chosun University Medical School, Gwangju, Korea

Corresponding author: Sung-Heui Shin, M.D. Research Center for Resistant Cells and Department of Microbiology, Chosun University Medical School, 375 Seoseok-dong, Dong-gu, Gwangju 501-759, Korea Tel: +82-62-230-6352, Fax: +82-62-233-6052 E-mail: shsin@chosun.ac.kr

∗ These authors contributed equally to this work.

∗ This study was supported by the National Research Foundation of Korea (NRF) grant funded by the Korean government through the Research Center for Resistant Cells (R13-2003-009).

      Revised 29 May 201017 May 2010       Accepted 12 July 2010

Copyright © 2010 Korean Society for Legal Medicine

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

We identified 6 sucrose-fermenting Vibrio vulnificus strains and examined their virulence characteristics. They were all encapsulated, motile, capable of producing toxins and utilizing transferrin-bound iron, cytotoxic to cultured cells, and virulent enough to kill mice. They could be definitely identified only by genetic identification methods such as PCR, and not by conventional culture-based identification methods such as API 20E (bioMérieux, France). These results indicate that it is essential to adopt genetic approaches as early as possible in order to avoid misdiagnosis of such strains, especially in clinical situations.

Keywords: Vibrio vulnificus, Sucrose, Thiosulfate-citrate-bile salts-sucrose agar, PCR, rDNA sequencing

REFERENCES

1.Jones MK., Oliver JD. Vibrio vulnificus: disease and pathogenesis. Infect Immun. 2009. 77:1723–33.
2.Harwood VJ., Gandhi JP., Wright AC. Methods for isolation and confirmation of Vibrio vulnificus from oysters and environmental sources: a review. J Microbiol Methods. 2004. 59:301–16.
3.Abbott SL., Janda JM., Johnson JA., Farmer III JJ. Vibrio and related organisms. Murray PR, Baron EJ, editors. Manual of clinical microbiology. 9th ed.Washington, DC: American Society of Microbiology;2007. p. 723–33.
4.Nagao M., Shimizu Y., Kawada Y., Baba H., Yamada K., Torii K, et al. Two cases of sucrose-fermenting Vibrio vulnificus infection in which 16S rDNA sequencing was useful for diagnosis. Jpn J Infect Dis. 2006. 59:108–10.
5.Reddy GP., Hayat U., Abeygunawardana C., Fox C., Wright AC., Maneval DR Jr, et al. Purification and determination of the structure of capsular polysaccharide of Vibrio vulnificus M06-24. J Bacteriol. 1992. 174:2620–30.
6.Dalsgaard A., Dalsgaard I., H⊘i L., Larsen JL. Comparison of a commercial biochemical kit and an oligonucleotide probe for identification of environmental isolates of Vibrio vulnificus. Lett Appl Microbiol. 1996. 22:184–8.
7.Hill WE., Keasler SP., Trucksess MW., Feng P., Kaysner CA., Lampel KA. Polymerase chain reaction identification of Vibrio vulnificus in artificially contaminated oysters. Appl Environ Microbiol. 1991. 57:707–11.
8.Shao CP., Hor LI. Metalloprotease is not essential for Vibrio vulnificus virulence in mice. Infect Immun. 2000. 68:3569–73.
9.Kim YR., Lee SE., Kook H., Yeom JA., Na HS., Kim SY, et al. Vibrio vulnificus RTX toxin kills host cells only after contact of the bacteria with host cells. Cell Microbiol. 2008. 10:848–62.
10.Kim CM., Park RY., Chun HJ., Kim SY., Rhee JH., Shin SH. Vibrio vulnificus metalloprotease VvpE is essentially required for swarming. FEMS Microbiol Lett. 2007. 269:170–9.
11.Kim CM., Park RY., Choi MH., Sun HY., Shin SH. Ferrophilic characteristics of Vibrio vulnificus and potential usefulness of iron chelation therapy. J Infect Dis. 2007. 195:90–8.
12.Kataoka E., Honma M., Ohnishi K., Sofuni T., Mizusawa H. Application of highly polymorphic DNA markers to the identification of HeLa cell sublines. In Vitro Cell Dev Biol. 1992. 28A:553–6.
crossref

Fig. 1.
Sucrose-fermenting Vibrio vulnificus strains forming yellow colonies on thiosulfate citrate bile salts sucrose (TCBS) agar. The M06-24/O strain (A), the 6 sucrose-fermenting V. vulnificus strains (Y-1 to Y-6; B), and other Vibrio species (C), V. alginolyticus ATCC17749 (Va) strain, V. cholerae Non O1 (Vc) strain, V. parahaemolyticus ATCC27519 (Vp) strain, and V. mimicus ATCC33653 (Vm) strain, were used. One colony of each strain grown on Heart Infusion agar was picked, streaked on TCBS agar, and incubated at 37°C for 24 hr.
kjlm-30-507f1.tif
Table 1.
Identification and virulence characteristics of sucrose-fermenting Vibrio vulnificus strains
Characteristics Y-1 Y-2 Y-3 Y-4 Y-5 Y-6 M06-24/O
Source E E C E E E C
TCBS Y Y Y Y Y Y G
API20E∗ 5147025 5047025 5047125 5247025 5147025 5047125 5247105
vvhA P P P P P P P
16S rDNA (%)† 100 99.8 99.9 100 99.9 99.9 100
Colony O O O O O O O
VvpE ++ ++ ++ ++ ++ ++ ++
VvhA ++ ++ ++ ++ ++ ++ ++
Swarming ++ ++ +‡ ++ +‡ ++ +++
TBI utilization ++ ++ ++ +‡ +‡ +‡ +++
Cytotoxicity (%)§ 101±5 41 ±4‖ 78±4‖ 15 ± 2‖ 32 ± 2‖ 101±3 109±7
Mouse lethality (hr) 2.7±0.2 15.8±6.9‖ 2.4±0.3 19.5 ± 1.4‖ 18.8 ± 0.0‖ 18.7 ± 0.0‖ 2.0±0.0

∗ The sixth one of the 7 digits indicates the ability to ferment sucrose;

† Sequence homology with the 16S rDNA of the V. vulnificus ATCC27562 strain

‡ P<0.05 in ANOVA on ranks; §Mean ±SE from triplicate experiments; ‖P<0.05 in one way ANOVA; Mean time required for killing the five mice.

Abbreviations: E, environmental; C, clinical; TCBS, thiosulfate citrate bile salts sucrose; Y, yellow; G, green; O, opaque; TBI, transferrin-bound iron; vvhA, Vibrio vulnificus cytolysin/hemolysin gene; P, positive PCR band with the expected molecular size; +, weak positive; ++, moderate positive; +++, strong positive.

TOOLS
Similar articles