미생물학회지 (Korean Journal of Microbiology)

  • 제54권1호
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  • Pages.53-59
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  • 2018
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  • 0440-2413(pISSN)
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  • 2383-9902(eISSN)
한국미생물학회 (The Microbiological Society of Korea)
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제주 양식장 유입수과 방출수에서 분리한 다제내성 Vibrio균 플라스미드 프로파일링

Plasmid profiling of multi-drug resistant Vibrio sp. isolated from influent and effluent water samples of fish farms in Jeju, South Korea

  • Farooq, Adeel (Faculty of Biotechnology, College of Applied Life Science, SARI, Jeju National University) ;
  • Unno, Tatsuya (Faculty of Biotechnology, College of Applied Life Science, SARI, Jeju National University)
  • 투고 : 2017.12.11
  • 심사 : 2018.01.08
  • 발행 : 2018.03.31
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초록

이 실험은 제주도내 양식장으로부터 유입수와 방출수의 다제내성(MDR)을 가진 Vibrio 균의 plasmid profiling을 위해 진행하였다. Plasmid profiling을 위해 사용한 다제내성을 가진 균주는 디스크 확산법을 통해 확인하였고, 유입수와 방출수으로부터 각각 150개의 Vibrio 균주를 분리하였다. 모든 다제내성 균주를 대상으로 plasmid profiling을 실시하였으며, 유입수와 비교하여 방출수에서 많은Vibrio 균이 열거되었다(유입수 39%, 방출수 61%). 방출수에서는 neomycin, sulfamethoxazole, amoxicillin 및 oxytetracycline의 내성을 가진 균주가 유의적으로 많은 것으로 확인되었고, 대조적으로 Vibrio 균주는 florfenicol, chloramphenicol, ciprofloxacin 및 nalidixic acid에 더 민감한 것으로 확인이 되었다. 99종의 다제내성 균주(유입수 39종과 방출수 60종) 중에서 총 58종(유입수 38종과 방출수 20종)이 1.7 kb에서 10 kb 이상의 플라스미드를 가지고 있는 것을 확인하였으며 플라스미드 크기마다 19가지의 다른 다제내성 패턴을 보였다. 6종의 유입수와 4종의 방출수에서 다제내성 균주는 특이적인 plasmid profile이 확인되었다. 방출수 샘플은 보다 많은 플라스미드를 가진 다제내성 Vibrio 균주와 다양한 plasmid profile들과 다제내성 패턴을 가지고 있었고 이는 양식장의 저장탱크가 항생제내성 유전자의 저장소 역할을 할 수 있다는 점을 시사한다. 양식장의 방출수에서 plasmid를 가진 다제내성 Vibrio 균주의 존재는 항생제 내성 유전자의 전파에 기여할 수 있으며 이로 인해 인간의 건강을 위협할 수 있다.

The objective of this study was to investigate the plasmid profiling of multi-drug resistant (MDR) Vibrio in influent (inflow) and effluent (discharged) water samples of fish farms in Jeju, South Korea. MDR isolates identified through disc diffusion susceptibility tests, were subjected to plasmid profiling. One hundred fifty Vibrio isolates were obtained from each influent and effluent water sample. All MDR isolates were subjected to plasmid profiling. Greater number of bacteria were enumerated from effluents (61%) comparing to influents (39%). High incidence of neomycin, sulfamethoxazole, amoxicillin and oxytetracycline resistance was observed among the isolates, which was higher in effluent samples. In contrast, Vibrio isolates were more susceptible to florfenicol, chloramphenicol, ciprofloxacin, and nalidixic acid. Among 99 (influent 39 and effluent 60) MDR isolates, a total of 58 (influent 38 and effluent 20) were found to bear plasmids ranging from 1.7 kb to >10 kb and showed 19 different antibiograms according to the size of plasmids. MDR isolates showed six and four distinct plasmid profiles in influent and effluent, respectively. Effluent samples contained more plasmid-carrying MDR Vibrio isolates with more diverse plasmid profiles and antibiograms, suggesting that fish farm tanks may serve as a reservoir of antibiotic resistance genes. The presence of plasmid-carrying MDR Vibrio isolates in fish farm effluent water may contribute to the dissemination of antibiotic resistance genes to the environments, which ultimately poses threat to human health.

키워드

참고문헌

  1. Ben-Haim, Y., Thompson, F.L., Thompson, C.C., Cnockaert, M.C., Hoste, B., Swings, J., and Rosenberg, E. 2003. Vibrio coralliilyticus sp. nov., a temperature-dependent pathogen of the coral Pocillopora damicornis. Int. J. Syst. Evol. Microbiol. 53, 309-315. https://doi.org/10.1099/ijs.0.02402-0
  2. Bien, T.L.T., Sato-Takabe, Y., Ogo, M., Usui, M., and Suzuki, S. 2015. Persistence of multi-drug resistance plasmids in sterile water under very low concentrations of tetracycline. Microbes Environ. 30, 339-343. https://doi.org/10.1264/jsme2.ME15122
  3. Buschmann, A.H., Tomova, A., Lopez, A., Maldonado, M.A., Henriquez, L.A., Ivanova, L., Moy, F., Godfrey, H.P., and Cabello, F.C. 2012. Salmon aquaculture and antimicrobial resistance in the marine environment. PLoS One 7, 26-28.
  4. Chelossi, E., Vezzulli, L., Milano, A., Branzoni, M., Fabiano, M., Riccardi, G., and Banat, I.M. 2003. Antibiotic resistance of benthic bacteria in fish-farm and control sediments of the Western Mediterranean. Aquac. 219, 83-97. https://doi.org/10.1016/S0044-8486(03)00016-4
  5. CLSI. 2015. Methods for antimicrobial dilution and disk susceptibility testing of infrequently isolated or fastidious bacteria, 3rd ed. CLSI document M 45. Clinical and Laboratory Standards Institute, Wayne, PA, USA.
  6. Garcia-Aljaro, C., Balleste, E., and Muniesa, M. 2017. Beyond the canonical strategies of horizontal gene transfer in prokaryotes. Curr. Opin. Microbiol. 38, 95-105. https://doi.org/10.1016/j.mib.2017.04.011
  7. Hormansdorfer, S., Wentges, H., Neugebaur-Buchler, K., and Bauer, J. 2000. Isolation of Vibrio alginolyticus from seawater aquaria. Int. J. Hyg. Environ. Health 203, 169-175. https://doi.org/10.1078/S1438-4639(04)70024-3
  8. Igbinosa, E.O. 2016. Detection and antimicrobial resistance of Vibrio isolates in aquaculture Environments: Implications for public health. Microb. Drug Resist. 22, 238-245. https://doi.org/10.1089/mdr.2015.0169
  9. Kang, C.H., Shin, Y.J., Jang, S.C., Yu, H.S., Kim, S.K., An, S., Park, K., and So, J.S. 2017. Characterization of Vibrio parahaemolyticus isolated from oysters in Korea: Resistance to various antibiotics and prevalence of virulence genes. Mar. Pollut. Bull. 118, 261-266. https://doi.org/10.1016/j.marpolbul.2017.02.070
  10. Letchumanan, V., Chan, K.G., and Lee, L.H. 2015. An insight of traditional plasmid curing in Vibrio species. Front. Microbiol. 6, 735.
  11. Luo, Y., Wang, Q., Lu, Q., Mu, Q., and Mao, D. 2014. An ionic liquid facilitates the proliferation of antibiotic resistance genes mediated by class i integrons. Environ. Sci. Technol. Lett. 1, 266-270. https://doi.org/10.1021/ez500103v
  12. Malla, S., Dumre, S.P., Shakya, G., Kansakar, P., Rai, B., Hossain, A., Nair, G.B., Albert, M.J., Sack, D., Baker, S., et al. 2014. The challenges and successes of implementing a sustainable antimicrobial resistance surveillance programme in Nepal. BMC Public Health 14, 269. https://doi.org/10.1186/1471-2458-14-269
  13. Manjusha, S. and Sarita, G.B. 2011. Plasmid associated antibiotic resistance in Vibrios isolated from coastal waters of Kerala. Int. Food Res. J. 18, 1171-1181.
  14. Manjusha, S. and Sarita, G.B. 2013. Characterization of plasmids from multiple antibiotic resistant Vibrios isolated from molluscan and crustacean of Kerala. Int. Food Res. J. 20, 77-86.
  15. Marie, B.C. 2005. Manual of antimicrobial susceptibility testing. American Society for Microbiology, Washington, DC, USA.
  16. Martinez-Urtaza, J., Blanco-Abad, V., Rodriguez-Castro, A., Ansede-Bermejo, J., Miranda, A., and Rodriguez-Alvarez, M.X. 2012. Ecological determinants of the occurrence and dynamics of Vibrio parahaemolyticus in offshore areas. ISME J. 6, 994-1006. https://doi.org/10.1038/ismej.2011.156
  17. Mirzoyan, N., Tal, Y., and Gross, A. 2010. Anaerobic digestion of sludge from intensive recirculating aquaculture systems: Review. Aquaculture 306, 1-6. https://doi.org/10.1016/j.aquaculture.2010.05.028
  18. Molina-Aja, A., Garcia-Gasca, A., Abreu-Grobois, A., Bolan-Mejia, C., Roque, A., and Gomez-Gil, B. 2006. Plasmid profling and antibiotic resistance of Vibrio strains isolated from cultured penaeid shrimp. FEMS Microbiol. Lett. 213, 8-13.
  19. Norman, A., Hansen, L.H., and Sorensen, S.J. 2009. Conjugative plasmids: vessels of the communal gene pool. Philos. Trans. R. Soc. B Biol. Sci. 364, 2275-2289. https://doi.org/10.1098/rstb.2009.0037
  20. Palmer, R., Kawai, K., and Kusuda, R. 1992. In vitro activity of quinolone antibacterials against selected fish pathogens. Fish Pathol. 27, 131-142. https://doi.org/10.3147/jsfp.27.131
  21. Pruden, A., Joakim Larsson, D.G., Amezquita, A., Collignon, P., Brandt, K.K., Graham, D.W., Lazorchak, J.M., Suzuki, S., Silley, P., Snape, J.R., et al. 2013. Management options for reducing the release of antibiotics and antibiotic resistance genes to the environment. Environ. Health Perspect. 121, 878-885. https://doi.org/10.1289/ehp.1206446
  22. Sandaa, R.A., Torsvik, V.L., and Goksoyr, J. 1992. Transferable drug resistance in bacteria from fish-farm sediments. Can. J. Microbiol. 38, 1061-1065. https://doi.org/10.1139/m92-174
  23. Schumann, M., Unger, J., and Brinker, A. 2017. Floating faeces: Effects on solid removal and particle size distribution in RAS. Aquacult. Eng. 78, 75-84. https://doi.org/10.1016/j.aquaeng.2016.10.007
  24. Serrano, P.H. 2013. Responsible use of antibiotics in aquaculture. Food Agriculture Organization, Rome, Italy.
  25. Silvester, R., Alexander, D., George, M., and Hatha, A.A.M. 2017. Prevalence and multiple antibiotic resistance of Vibrio coralliilyticus, along the Southwest Coast of India. Curr. Sci. 112, 1749-1749. https://doi.org/10.18520/cs/v112/i08/1749-1755
  26. Suzuki, S., Ogo, M., Koike, T., Takada, H., and Newman, B. 2015. Sulfonamide and tetracycline resistance genes in total- and culturable-bacterial assemblages in south african aquatic environments. Front. Microbiol. 6, 1-8.
  27. Thompson, F.L., Gevers, D., Thompson, C.C., Dawyndt, P., Hoste, B., Munn, C.B., Swings, J., and Naser, S. 2005. Phylogeny and molecular identification of Vibrios on the basis of multilocus sequence analysis phylogeny and molecular identification of Vibrios on the basis of multilocus sequence analysis. Appl. Environ. Microbiol. 71, 5107-5115. https://doi.org/10.1128/AEM.71.9.5107-5115.2005
  28. Vezzulli, L., Chelossi, E., Riccardi, G., and Fabiano, M. 2002. Bacterial community structure and activity in fish farm sediments of the Ligurian sea (Western Mediterranean). Aquacult. Int. 10, 123-141. https://doi.org/10.1023/A:1021365829687
  29. Yang, J.H., Mok, J.S., Jung, Y.J., Lee, K.J., Kwon, J.Y., Park, K., Moon, S.Y., Kwon, S.J., Ryu, A.R., and Lee, T.S. 2017. Distribution and antimicrobial susceptibility of Vibrio species associated with zooplankton in coastal area of Korea. Mar. Pollut. Bull. 125, 39-44. https://doi.org/10.1016/j.marpolbul.2017.07.054
  30. Yoo, M.H., Huh, M.D., Kim, E.H., Lee, H.H., and Jeong, H.D. 2003. Characterization of chloramphenicol acetyltransferase gene by multiplex polymerase chain reaction in multidrug-resistant strains isolated from aquatic environments. Aquaculture 217, 11-21. https://doi.org/10.1016/S0044-8486(02)00169-2
  31. You, K.G., Bong, C.W., and Lee, C.W. 2016. Antibiotic resistance and plasmid profiling of Vibrio spp. in tropical waters of Peninsular Malaysia. Environ. Monit. Assess. 188, 1-15. https://doi.org/10.1007/s10661-015-4999-z
  32. Zanetti, S., Spanu, T., Deriu, A., Romano, L., Sechi, L.A., and Fadda, G. 2001. In vitro susceptibility of Vibrio spp. isolated from the environment. Int. J. Antimicrob. Agents 17, 407-409.