Isolation and Characteristics of Yersinia spp. from Mineral Spring Waters

먹는 샘물에서 분리한 Yersinia spp.의 분리 및 특성

  • 이택수 (강원도 보건환경연구원) ;
  • 박부길 (강원도 보건환경연구원) ;
  • 오덕환 (강원대학교 식품생명공학부)
  • Published : 2001.10.01

Abstract

A total of 277 mineral spring water samples in Kangwon province from 1999 to 2000 were analyzed for the presence of Yersinia spp. by the conventional Food and Drug Administration protocol, and presumptive strains were identified by morphological, cultural and biochemical tests according to Bergey’s manual. Also, the biotypes, serotypes, and susceptibility to 12 antibiotics were tested. Among the total 277 mineral spring water samples, 40 samples (14.4%) were found to be contaminated with Yersinia species. Among the 40 strains of Yersinia spp. isolates, 33 strains (82.5%) for Yersinia enterocolitica, 4 strains (10%) for Yersinia frederiksenii, 2 strains (5%) for Yersinia intermedia, and 1 strain (2.5%) for Yersinia sakazaki were identified, respectively. Of 40 Yersinia spp. isolates, Yersinia enterocolitica (82.5%) was the most predominant species in the mineral spring water samples compared to other Yersinia species. Compared to direct culture method after KOH treatment and KOH treatment method after cold enrichment for better isolation ratio of according to comparision of Yersinia species, the detection ration (18.5%) of KOH treatment method after cold enrichment was about 3 times better than that (6.1%) of direct culture method after KOH treatment. According to serotypes of Y. enterocolitica isolates, O : 5 (12.9%) was the most predominant and followed by O : 3 (9.7%), O : 8 (6.5%), and O : 9 (3.2%), and others. For biotypes of Y. enterocolitica isolates, 1A (71.0%) was the most predominantly abundant and followed by 3A (12.9%), 3B (9.7%), 1B (3.2%) and 5 (3.2%). Also, an antibiotic susceptibility test showed that Yersinia spp. isolates were very susceptible to the antibiotics tested, but they were very strongly resistant to ampicillin, cephalothin and carbenicillin.

최근에 음용이 증가하고 있는 강원도지역 먹는 샘물로부터 Yersinia spp.의 부포현황을 조사하기 위하여 1999년과 2000년에 강원지역 18개 시군 전역에 산재한 공동샘터수 277건을 수거한 후 미국 FDA방법과 Bergey’s manual을 참조하여 형태학적, 배양학적, 생화학적방법으로 균을 분리동정하였으며 분리균의 생물형, 혈청형, 항균제 감수성조사 및 효율적인 균리방법에 대한 조사를 실시하였다. 샘터수 277건 중에 40건(14.4%)에서 Yersinia spp.가 분리되었으며, 균종별 분리빈도는 40건중에서 Y. enterocolitica 33주(82.5%)로 가장 많이 분리되었으며, Y. frederiksenii 4주(10%), Y. in-termedia 2주(5%), Y.sakazaki 1주(2.5%)순으로 분리되었다. 지역별로 Y. enterocolitica 분리실태를 보면 철원군(5주), 인제군(5주) 등 11개 군지역에서 평균 2.6주가 분리되었고, 태백시(2주), 춘천시(1주) 등 3개 시지역에서 평균 1.3주가 분리되어 군소재 먹는 샘물이 도시지역 먹는 샘물에 비해 2배 정도 높은 균검출율을 나타내었다. 배양방법에 따른 Y. enterocolitica 분리율을 비교하면 시료를 KOH처리하여 직접 CIN 배지에 분리한 직접배양법에서는 분리율이 6.1%이었고, 시료를 PSBB배지에 1$0^{\circ}C$, 10일간 저온배양 후 KOH로 처리하여 CIN배지에 분리한 저온증균배양법에서 18.5%의 분리율을 나타내어 저온증균법이 직접법에 비해 3배 높은 균검출 효과를 나타내었다. 한편, Y. enterocolitica 33주에 대한 아형(subtype) 조사 결과 생물형 분포는 1A형이 22주(71.0%)로 가장 많았고 3A형 4주(12.9%), 3B형 3주(9.7%), 1B형과 5형이 각각 1주(3.2%)씩이었으며, 혈철형 분포는 O : 5형이 4주(12.9%)로서 가장 많았으며 O : 3형 3주(9.7%), O : 8형 2주(6.5%) 및 O : 9형 1주(3.2%)이었고 나머지 21주(67.7%)는 혈청형을 확인할 수 없었다. 또한, 분리된 Y. enterocolitica의 12종 항생제에 대한 감수성 시험결과는 ampicillin, cephalothin, carbenicillin에 각각 70%, 70%, 52%의 저항성을 보였고 나머지 nalidixic acid, streptomycin, tetracycline, tobramycin, chloramphenicol, kanamycin gentamycin, trimethoprim, ciprofloxacin에 대하여는 대부분 감수성을 나타내었으며, ampicillin, cephalothin, carbenicillin 등 3종 항생제에 다제내성을 보인 경우는 39%이었다.

Keywords

References

  1. Bergey's manual of Systemic Bacteriology v.1 Genus Yersinia Noel, R. K.;John, G. H.
  2. The Genus Yersinia, Identification of enterobacteriaceae(4th ed.) Edward;Ewing's
  3. Color atalas and textbook of diagnostic microbiology(3th ed.) Koneman, E. W.;Allen, S. D.;Janda, W. M.;Schreckenberger, P. C.;Winn, W. C.
  4. Foodborne Bacterial Pathogens(1st ed.) Y. enterocolitical and Y. pseudotuberculosis Schiiemann, D. A.
  5. J. Food Sci. v.45 Effects of pH and sodium chloride on Y. enterocolitica growth at room and refrigeration temperatures Stern, N. J.;Pierson, M. D.;Kotula, A. W.
  6. J. Appl. Bacteriol. v.52 A review of Yersinia enterocolitica Swaminathan, B.;Harmon, M. C.;Mehlman, I. J.
  7. Palque and the Yersinia Infections Bulter, T.
  8. Korean J. Vet. Res. v.34 Biotype, serotype and antibiotics suscepibility of Yersinia enterocolitica isolated from zoo animals Park, S. G.;Youn, E. S.;Kim, E. J.
  9. J. Korean Soc. Microbiol. v.25 Yersinia species and serogroups and biotypes of Yersinia enterocolitica isolate from animals (pigs and dogs) in Koera Choi, C. S.;Park, S. G.;Youn, Y. D.;Chung, S. I.;Yang, Y. T.
  10. Korean J. Food Sci. Technol. v.31 Characteristics of Yersinia enterocolitica isolated from foods Lim, S. Y.;Lee, D. H.;Park, S. H.;Kim, C. M.
  11. Appl. Environ. Microbiol. v.50 Isolation of Y. enterocolitica and related species from food in France Delmas, C. L.;Vidon, D. J. M.
  12. Int. J. Food Microbiol. v.21 Incidence of Yersinia spp. in foog in Sao Paulo, Brazil Tassinari, a.;Franco, B. D. G.;Landgraf, M.
  13. Ph.D Dissertation, Dankook University Biological, serological and molecular typing of Y. enterocolitica isolated from spring water in seoul Lee, Y. K.
  14. Acta Path Micribiol. Scand Sec. B. v.85 Y. enterocolitica and Yersinia like microbes isolated from mammals and water in Norway and Den mark Georg, K.
  15. N. Eng. J. Med. v.321 Medical progress-(Yersinia enterocolitica) Cover, T. L.;Aber, R. C.
  16. Int. J. Food Microbiol. v.12 Yersinia enterocolitica in food hygiene Kapperud, G.
  17. Int. J. Food Microgiol. v.13 The microbiological quality of drinking water supplir of lzmir city: The incidence of Yersinia enterocolitica AktugGonul, S.;Karapmar, M.
  18. Appl. Environ. Microbiol. v.34 Isolation of Yersinia enterocolitica from well water and growth in distilled water Highsmith, A. K.;Feeley, J. C.;Skaliy, P.;Wells, J. G.;Wood, B. T.
  19. J. Korean Ped. Assoc. v.33 An outbreak of Yersinia pseudotuberculosis Joo, H. J.;Chung, K. H.;Kim, Y. M.;Kim, S. G.;Park, M. S.;Chang, J. K.;Shin, S. W.
  20. Bacteriological Analycical Manual(7th ed.) Y. enterocolitica and Y. pseudotubrculosis Stephan, D.;Weagant, P. F.;John, T. S.
  21. Curr. Microbiol. v.11 Biochemical heterogenecity of serotype O; 3 strains of 700 Yersinia strains isolates from humans, other mammals, flies, animal feed, and river water Fukushima, H.;Tsubokura, M.;Oksuki, K.;Kawaoka, Y.
  22. Am. J. Clin. Pathol. v.45 Antibiotiotic susceptibility testing by a standardized single disk method Bauer, A. W.;Kirby, W. M. M.;Sherris, J. C.
  23. Performance Standards for Antimicrobial Disk Susceptibility Tests (Approved Standard, M2-M5) National Committee for Clinical Laboratory Standards
  24. The Report of Public Health and Environment Institute of Taejon Metropolitan City v.2 A research on the Y. enterocolitica isolated from mineral spring water in Taejon metropolitan city Nam, S. W.;Kwon, E. R.;Park, S. K.;Bing, S. H.;Kim, J. Y.
  25. The Report of Public Health and Environment Institute of Kyongsangnamdo v.5 Studies on isolationand pathogenice of Yersinia from mineral spring water in Kyengnam area Choi, S. M.;Chung, J. H.;Cha, H. G.;Kang, Y. H.;Kim, Y. R.;Lee, K. H.
  26. Appl. Environ. Microbiol. v.32 Recovery of Y. enterocolitica from streams and lacks of Califonia Sydney Harvey, J. R.;Greenwood, M. J.;Robert, M. A. H.
  27. The Report of Public Health and Environment Institute of Seoul v.12 Studies on comparsion of detection method for Yersinia spp. and antibiotic resistene of isolates Lee, H.;Choi, S. M.;Ham, H. J.;Lee, Y. G.;An, M. J.;Choi, C. M.;Oh, S. K.
  28. J. Korean Soc. Microbiol. v.28 Biotype, serotype and antimicrobial susceptibility of Yersinia enterocolitica isolated from cattle Park, S. G.;Choi, S. M.;Oh, Y. H.;Choi, C. S.