DOI QR코드

DOI QR Code

Prevalence of Toxin Genes and Antibiotic Resistance Profiles of Vibrio vulnificus strains isolated from Jeju Island

제주도에서 분리된 비브리오패혈증균의 독소 유전자 분포 및 항생제 내성

  • 강은옥 (제주특별자치도 보건환경연구원) ;
  • 조만재 (제주특별자치도 보건환경연구원) ;
  • 허예슬 (제주특별자치도 보건환경연구원) ;
  • 고은아 (제주특별자치도 보건환경연구원)
  • Received : 2023.07.07
  • Accepted : 2023.09.22
  • Published : 2023.10.30

Abstract

Vibrio vulnificus, the most fatal waterborne and foodborne pathogens of 50% fatality rate in the world, is common in seawater and occurs particularly in warmer months. In this study, we investigated the toxin genes using reverse transcription-polymerase chain reaction (RT-PCR), antibiotic resistance status using Vitek, and genetic characteristics using pulsed-field gel electrophoresis (PFGE) of different V. vulnificus strains isolated from the Jeju Island seawater, distribution fishery products, and fish tanks. We examined a total of 487 samples and isolated a total of 46 strains (including overlapping strains) of V. vulnificus, 44 strains from seawater and 1 strain each from fishery products and fish tank. We detected toxin gene vvhA in all 46 strains and rtxA, viu in 8 strains (17.4%) and 9 strains (19.6%) strains, respectively. Antibiotic resistance tests indicated 100% resistance to cefoxitin antibiotics. The PFGE analysis of the 46 strains identified a total of 6 types showed 100% homology and the degree of similarity was 81.3-98.0%; however, there were no similarity between the regions and samples. These results indicate that V. vulnificus isolated from the seawater, fishery products, and fish tanks should be continuously monitored as cases of food poisoning caused by V. vulnificus with toxin genes have been reported in Jeju Island.

비브리오패혈증균은 세계에서 치사율이 50%에 달하는 가장 치명적인 수인성식품매개 병원균으로 해수에서 흔히 있으며, 특히 따뜻한 계절에 발생한다. 본 연구는 제주도의 해수, 유통 수산물, 수족관물에서 분리한 비브리오패혈증균에 대해서 RT-PCR을 이용한 독소 유전자, Vitek을 이용한 항생제 내성, PFGE를 이용한 유전적 특성을 조사하였다. 총 487개의 시료를 조사한 결과 비브리오패혈증균 46주(중복 균주 포함)가 해수에서 44주, 유통수산물에서 1주, 수족관물에서 1주 분리되었다. rtxA, viu와 같은 독소 유전자는 각각 8주(17.4%), 9주(19.6%) 검출되었고, vvhA와 같은 독소 유전자는 모든 균주에서 검출되었다. 항생제 내성 실험결과 cefoxitin 항상제에 대해서 100% 내성이 나타났다. 비브리오패혈증균 46주에 대한 PFGE 분석 결과 총 6유형이 100% 상동성을 보였고, 유사도는 81.3-98.0%로 나타났다. 수산물과 수족관물에서 분리된 비브리오패혈증균은 해수와의 상동성 결과 유사도는 불일치로 나타났고 지역과 시료 사이에는 유사성이 없었다. 독소 유전자를 가진 비브리오패혈증균에 의한 식중독 환자가 제주도에서 발생한 점을 고려해볼 때, 해수, 유통 수산물, 수족관물에서 분리한 비브리오패혈증균에 대한 모니터링이 지속되어야 할 것으로 보인다.

Keywords

Acknowledgement

본 연구는 제주특별자치도 보건환경연구원 연구사업의 지원으로 수행되었기에 감사드립니다.

References

  1. Baker-Austin, C., Oliver, J.D., Vibrio vulnificus: new insights into a deadly opportunistic pathogen. Environ. Microbiol., 20, 423-430 (2018).  https://doi.org/10.1111/1462-2920.13955
  2. Lee, H.A., Ko, Y.E., Choi, J.H., Lee, D.Y., Yeo, S.S., Park, J.J., Lee, M.Y., Choi, J.H., Park, J.H., Distribution and characterization of Vibrio vulnificus isolated in coastal areas of Chungcheongnam-do province. J. Environ. Health. Sci., 47, 479-485 (2021).  https://doi.org/10.5668/JEHS.2021.47.5.479
  3. Yoon, Y.H., Park, S., Kim, J.Y., Lee, Y.J., Jeon, D.Y., Choi, G.C., Park, J.S., Kim, J.B., Prevalence of toxin genes and profiles of antibitoc resistance in Vibrio vulnificus isolates from fish, fish Tanks, and patients. J. Food Hyg. Saf., 35, 6-12 (2020).  https://doi.org/10.13103/JFHS.2020.35.1.6
  4. Yang, S.J., Kim, H.S., Kwon, J.Y., Chang, S.H., Kim, Y.M., Ecological characteristics of Vibrio vulnificus in estuary of Kum River. J. Food Hyg. Saf., 10, 53-59 (1995). 
  5. Oh, H.K., Jeong, H.J., Kim, Y.M., Distribution and molecular characteristics of Vibrio vulnificus Isolated from seawater along the Gadeok Island coast. Korean J. Fish. Aquat. Sci., 53, 688-693 (2020). 
  6. Oh, B.Y., Kim, J.H., Gong, Y.W., Jegal, S., Kim, H.Y., Lee, M.Y., Hwang, K.W., Koh, Y.J., Lee, J.M., Go, J.M., Kim, Y.H., Characteristics of Vibrio vulnificus Isolated in Incheon. Korean J. Microbiol., 43, 256-263 (2007). 
  7. Kim, H.S., Heo, S.T., Bae, I.G., Lim, M.H., Kim, J.H., Yun, S.E., Kim, S.J., A case of acute bacterial meningitis caused by Vibrio vulnificus. Korean J. Med., 78, 523-526 (2010). 
  8. Goo, J.S., Kim, D.W., Han, K.S., Suk, J.S., Park, M.H., Kim, S.I., Lactose fermenting vibrio (Vibrio vulnificus) septicemia: report of five cases. Korean J. Pathol., 16, 463-468 (1982). 
  9. Korea Disease Control and Prevention Agency (KDCA), (2023, June 24). Infectious disease. Retrieved from https://npt.kdca.go.kr/npt/biz/npp/ist/bass/bassDissStatsMain.do 
  10. Yang, S.J., Shin, J.H., Cho, D., Kee, S.J., Shin, M.G., Suh, S.P., Yang, D,W., Identification of Vibrio vulnificus by the Microscan and the Vitek II systems. Korean J. Lab. Med., 25, 33-38 (2005). 
  11. Lee, C.Y., Current insights into sepsis treatments. Acute Crit. Care, 25, 207-211 (2010). 
  12. Cabello, F.C., Godfrey, H.P., Tomova, A., Ivanova, L., Dolz, H., Millanao, A., Buschmann, A.H., Antimicrobial use in aquaculture re-examined: its relevance to antimicrobial resistance and to animal and human health. Environ. Microbiol., 15, 1917-1942 (2013).  https://doi.org/10.1111/1462-2920.12134
  13. Song, J.W., Jeong I.H., Park, S.H., Yun, S.J., Hur, E.S., Kim, Y.S., Characterization of Vibrio vulnificus isolated from the west coastal area of Gyeonggi-do. Korean J. Microbiol., 57, 197-203 (2021). 
  14. Li, G., Wang, M.Y., The role of Vibrio vulnificus virulence factors and regulators in its infection-induced sepsis. Folia Microbiol., 65, 265-274 (2020).  https://doi.org/10.1007/s12223-019-00763-7
  15. Lo, H.R., Lin, J.H., Chen, Y.H., Chen, C.L., Shao, C.P., Lai, Y.C., Hor, L.I., RTX toxin enhances the survival of Vibrio vulnificus during infection by protecting the organism from phagocytosis. J. Infect. Dis., 203, 1866-1874 (2011).  https://doi.org/10.1093/infdis/jir070
  16. Lopez-Canovas, L., Benitez, M.B.M., Isidron, J.A.H., Soto, E.F., Pulsed field gel electrophoresis: past, present, and future. Anal. Biochem., 573, 17-29 (2019).  https://doi.org/10.1016/j.ab.2019.02.020
  17. Jo, M.H., Kim, S.G., Kim, Y.H., Kim, S.T., Eom, H.J., Jang, Y.S., Ko, Y.H., Genotyping of Brucella abortus isolated in Gyeongbuk province by PFGE. Korean J. Vet. Serv., 32, 257-264 (2009). 
  18. Lee, D.S., Shin, H.Y., Park, K.S., Shin, I.S., Distribution of Vibrio parahaemolyticus and Vibrio vulnificus in seawater and shellfish at Gomso Bay. Korean J. Fish. Aquat. Sci., 52, 114-120 (2019). 
  19. Ministry of Food and Drug Safety (MFDS), Korea food code, Cheongju, Korea (2021). 
  20. Clinical and Laboratory Standards Institute (CLSI), Performance standards for antimicrobial susceptibility testing, 33rd ed., Wayne, PA, USA, 30-52 (2018). 
  21. Korea Disease Control and Prevention Agency (KDCA), PFGE standard test method, Cheongju, Korea (2022). 
  22. Baker-Austin, C., Oliver, J.D., Vibrio vulnificus: new insights into a deadly opportunistic pathogen. Environ. Microbiol., 20, 423-430 (2018).  https://doi.org/10.1111/1462-2920.13955
  23. Kim, D.M., Hong, S.J., Vibrio vulnificus sepsis. Korean J. Med., 82, 671-679 (2012).  https://doi.org/10.3904/kjm.2012.82.6.671
  24. Go, Y.J., Jang, J.S., Relationships of pathogenic vibrios and environmental factors affecting their occurrence in the seawater of incheon coastal Area. Korea J. Food. Nutr., 3, 414-420 (2013). 
  25. Noh, Y.M., Kim, H.J., Seo, S.R., Lee, D.H., 2020 epidemiologic analysis of Vibrio species patients and deaths. Public Health Wkly. Rep., 14, 1837-1849 (2021). 
  26. Choi, Y.M., Bae, J.H., Analysis of genetic diversity of Salmonella Typhi chromosomal genetic diversity analysis using pulsed-field electrophoresis. Korean J. Clin. Pathol., 18, 343-351 (1998). 
  27. Tamplin, M.L., Jackson, J.K., Buchrieser, C., Murphhree, R.L., Portier, K.M., Gangar, V., Miller, L.G., Kaspar, C.W., Pulsed-field gel electrophoresis and ribotype profiles of clinical and environmental Vibrio vulnificus isolates. Appl. Environ. Microbiol., 62, 3572-3580 (1996). https://doi.org/10.1128/aem.62.10.3572-3580.1996