Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
권호 표지

The Types of Extended-Spectrum $\beta$-Lactamase (ESBL) Produced by Enteric Bacteria, Klebsiella pneumoniae and Escherichia coli Isolated from Sewage of Wastewater Treatment Plant at Minragdong in Busan, Korea

부산 민락동 오수처리장에서 분리된 장내세균 Klebsiella pneumoniae와 Escherichia coli가 생성한 광범위 베타 락탐(Extended-Spectrum $\beta$-Lactamase, ESBL) 분해효소의 유형

  • Lee, Hun-Ku (Department of Microbiology, College of Natural Sciences, Pukyong National University)
  • 이훈구 (부경대학교 자연과학대학 미생물학과)
  • Received : 2009.05.21
  • Accepted : 2009.06.23
  • Published : 2009.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

This study was performed to investigate the type of extended-spectrum $\beta$-lactamases (ESBL) produced by bacteria isolated from the sewage of wastewater treatment plant at Minragdong, Suyong-gu in Busan. The facility is located at sushi restaurants and guides its drain water to the wastewater treatment plant at Yonghodong, Nam-gu in Busan. Samples were collected on January, 2009. A total of 19 strains were selected as potential ESBL positive strains through a double disk synergy test. On the basis of the results from biochemical tests including indole, methyl-red, Voges-Proskauer, Simmon's citrate, decarboxylase-dihydrolase and sugar-fermentation tests, the 19 strains were identified with 16 strains of Escherichia coli and 3 strains of Klebsiella pneumoniae. Out of 19 strains, 4 transconjugants against Escherichia coli J53, which is sodium azide resistant recipient strain, were obtained. The plasmids isolated from transconjugants were used for PCR analysis. The type of each extended-spectrum $\beta$-lactamase (ESBL) produced by the strains was determined on the basis of isoelectric focusing analysis and DNA sequencing. The results indicated that the types of ESBL from Klebsiella pneumoniae were SHV-12 (3 strains), and Escherichia coli was SHV-12/TEM-1 (1 strain), respectively.

본 논문은 부산소재 하수처리장 중계시설인 민락오수처리장의 하수에서 광범위 베타 락탐 분해효소(extendedspectrum $\beta$-lactamase, ESBL) 생성균주를 분리 동정하고 이들이 생성한 ESBL 유형을 조사하였다. 민락오수처리장은 부산 남구 수영구의 민락동 일대에 밀집한 횟집의 하수를 모아서 남구 용호동에 있는 하수종말처리장으로 중계시키는 시설이다. 2009 년 1 월 하수 검체로부터 항균제 이중 디스크 확산 시험(double disk synergy test)에 양성반응을 나타낸 19 균주를 1차 시험균주로 선택하였다. 인돌생성 시험 methyl-red, Voges-Proskauer, Simmon's citrate, decarboxylase-dihydrolase 및 당 발효능 생화학 시험을 통하여 Klebsiella pneumoniae (3 균주)와 Escherichia coli (16 균주)가 동정되었다. 이 균들을 공여균주로, Escherichia coli J53 (sodium $azide^r$)를 피전달균주로 하여 접합시험을 실시하여 plasmid로 매개된 4 균주 접합체를 얻었다. 이들로부터 plasmid를 추출하여 PCR로 유전자 증폭을 시켜 유전자를 분석하고 등전점을 조사한 결과 Klebsiella pneumoniae에서 생성된 ESBL 유형은 모두 SHV-12 (3 균주)였고, Escherichia coli에서 생성된 ESBL 유형은 SHV-12/TEM-1 (1 균주)였다.

Keywords

References

  1. 이경원, 조성란, 이창숙, 정윤섭, 권오헌. 1994. Extended broad-spectrum $\beta$-lactamase 생성 Escherichia coli와 Klebsiella pneumoniae. 감염 26, 341-348
  2. Ambuler, R.P., A.F.W. Coulson, N.M. Frere, J.M. Ghuysen, B. Joris, M. Forsman, R.C. Lebversque, G. Tiraby, and S.G. Waley. 1991. A standard numbering scheme for class A $\beta$-lactamases. Biochem. J. 276, 269-272 https://doi.org/10.1042/bj2760269
  3. Brinas, L., M. Zarazaga, Y. Saenz, F. Ruiz-Larrea, and C. Torres. 2002. $\beta$-Lactamases in ampicillin-resistant Escherichia coli isolates from foods, humans, and haelthy animals. Antimicrob. Agents Chemother. 46, 3156-3163 https://doi.org/10.1128/AAC.46.10.3156-3163.2002
  4. Bush, K. 1989. Characterization of $\beta$-lactamases. Antimicrob. Agents Chemother. 33, 259-263 https://doi.org/10.1128/AAC.33.3.259
  5. Bush, K., G.A. Jacoby, and A.A. Medeiros. 1995. A functional classification scheme for $\beta$-lactamases and its correlation with molecular structure. Antimicrob. Agents Chemother. 39, 1211-1233 https://doi.org/10.1128/AAC.39.6.1211
  6. Bush, K. and G. Jacoby. 1997. Nomenclature of TEM $\beta$-lactamases. Antimicrob. Agents Chemother. 39, 1-3 https://doi.org/10.1093/jac/39.1.1
  7. Ewing, W.H. 1986. The genus Klebsiella. In W.H. Ewing, Edwards and Ewing's Identification of Enterobacteriaceae. Elsevier, N.Y., USA.
  8. Jacoby, G.A. 1994. Genetics of extended-spectrum beta-lactamases. Eur. J. Clin. Microbiol. Infect. Dis. 13, 2-11 https://doi.org/10.1007/BF02390679
  9. Jacoby, G.A. and I. Carreras. 1990. Activities of $\beta$-lactam antibiotics against Escherichia coli strains producing extended-spectrum $\beta$-lactamases. Antimicrob. Agents Chemother. 34, 859-862
  10. Jeong, Y.S., J.C. Lee, H.Y. Kang, H.S. Yu, E.Y. Lee, C.H. Choi, S.H Tae, Y.C. Lee, D.T. Cho, and S.Y. Seol. 2003. Epidemiology of nalidixic acid resisitance and TEM-1-and TEM-52-mediated ampicillin resistance of Shigella sonnei isolates obtained in Korea between 1980 and 2000. Antimicrob. Agents Chemother. 47, 3719-3723 https://doi.org/10.1128/AAC.47.12.3719-3723.2003
  11. Kim, J., Y. Kwon, H. Pai, J.W. Kim, and D.T. Cho. 1998. Survey of Klebsiella pneumoniae strains producing extended-spectrum $\beta$-lactamase: prevalence of SHV-12 and SHV-2a in Korea J. Clin. Microbiol. 36, 1446-1449
  12. Lee, H.K. 2006. Typing of extended-spectrum $\beta$-lactamase (ESBL) producing Enterobacteriaceae isolated from slaughterhouse in Pusan, Korea. Kor. J. Microbiol. 42, 125-130
  13. Lee, H.K., H.J. Kim, and G.D. Kim. 2007. Typing of extendedspectrum $\beta$-lactamases of Escherichia coli and Klebsiella pneumoniae isolated from rivers in Pusan, Korea. Kor. J. Microbiol. 43, 116-123
  14. Medeiros, A.A. 1993. Nosocomial outbreaks of multiresistant bacteria: extended- spectrum beta-lactamases have arrived in North America. Ann. Intern. Med. 119, 428-430 https://doi.org/10.7326/0003-4819-119-5-199309010-00015
  15. National Committee for Clinical Laboratory Standards. 1997. Performance standards for antimicrobial disk susceptibility tests. NCCLS document M2-A6. NCCLS, Wayne, Pa, USA.
  16. Nueesch-Inderbinen, M.T., F.H. Kayser, and H. Haechlier. 1997. Survey and molecular genetics of SHV $\beta$-lactamases in Enterobacteriaceae in Switzerland: two novel enzymes, SHV-11 and SHV-12. Antimicrob. Agents Chemother. 41, 943-949
  17. Pai, H.J., S. Lyu, J.H. Lee, J. Kim, and Y. Kwon. 1999. Survey of extended-spectrum $\beta$-lactamase in clinical isolates of Escherichia coli and Klebsiella pneumoniae: prevalence of TEM-52 in Korea. J. Clin. Microbiol. 37, 1758-1763
  18. Philippon, A., R. Labia, and G. Jacobi. 1989. Extended-spectrum $\beta$-lactamases (minireview). Antimicrob. Agents Chemother. 33, 1131-1136 https://doi.org/10.1128/AAC.33.8.1131
  19. Silva, J., R. Gatoca, C. Auguilar, Z. Becerra, U. Garza-Ramos, M. Velzqez, G. Miranda, B. Leaos, F. Solrzano, and G. Echniz. 2001. Outbreak of infection with extended-spectrum $\beta$-lactamase producing Klebsiella pneumoniae in a Mexican Hospital. J. Clin. Microbiol. 39, 3193-3196 https://doi.org/10.1128/JCM.39.9.3193-3196.2001
  20. Teshager, T., L. Domnguez, M.A. Moreno, Y. Sanz, C. Torres, and S. Cardeosa. 2000. Isolation of an SHV-12 $\beta$-lactamase-producing Escherichia coli strain from a dog with recurrent urinary tract infections. Antimicrob. Agents Chemother. 44, 3483-3484 https://doi.org/10.1128/AAC.44.12.3483-3484.2000