Detection of CTX-M and Clonal Analyses using MLST in Cefotaxime Resistant $Escherichia$ $coli$ Isolated from the Han-River, Korea

  • Son, Gun-Woo (Department of Life Science, Sangmyung University) ;
  • Kim, Mal-Nam (Department of Life Science, Sangmyung University)
  • Received : 2011.10.04
  • Accepted : 2011.11.10
  • Published : 2011.11.30

Abstract

Bacteria resistant to various antibiotics have recently become an issue of the utmost importance. Resistant strains are not uncommon, even in municipal drinking water sources. The health threat posed by resistant, pathogenic bacteria has serious ramifications for both public health and agriculture. In this study, we isolated antibiotic resistant bacteria from water samples from the Han River, Korea, which is contaminated by the wastewater from many industrial complexes, hospitals, agricultural and animal husbandry estates, and from wastewater treatment facilities. We determined the degrees of resistance to various antibiotics exhibited by the isolated strains. The similarities between the isolated $E.$ $coli$ strains were examined, using the pulsed field gel electrophoresis and multilocus sequence typing, in order to trace their origins and to explore the syntechnic adaptations and pathogenicity of the various strains and relate these to their genetic sequence. A total of 25 $E.$ $coli$ strains were isolated from six stations along the Han River. All the 25 strains exhibited resistance to ampicillin. We also investigated resistance to amoxicillin, clavulanic acid, cefazolin, cofoxitin, cefotaxime, cefpodoxime, ceftriaxone, cefepime, nalidixic acid, aztreonam, ciprofloxacin, streptomycin, gentamicin, chloramphenicol and imipenem. Based on the ESBL detection, 14 strains belonged to the ESBL producing strains. The number of the clonal complex producing strains was 5 among the 14 isolated strains. The 5 strains were included in the 168, 23, 38, 469, 156 clonal complex, respectively. The rest 9 strains were not included in the clonal complex, but showed independent STs.

Keywords

References

  1. Achour NB, PS Mercuri, P Power, C Belhadj, MB Moussa, M Galleni and O Belhadj. 2009. First detection of CTX-M-28 in a Tunisian hospital from a cefotaxime-resistant Klebsiella pneumoniae strain. Pathol. Biol. 57:343-348. https://doi.org/10.1016/j.patbio.2008.07.016
  2. Armstrong JL, JJ Calomiris and RG Seidle. 1982. Selection of antibiotic resistant standard plate count bacteria during water treatmen. Appl. Environ. Microbiol. 44:308-316.
  3. Bonnet RC, D Champs, D Sirot, C Chanal, R Labia and J Sirot. 1999. Diversity of TEM Mutants in Proteus mirabilis. Antimicrob. Agents Chemother. 43:2671-2677.
  4. Bonnet R. 2004. Growing group of extended-spectrum $\beta$-lactamases: the CTX-M enzymes. Antimicrob. Agents Chemother. 48:1-14. https://doi.org/10.1128/AAC.48.1.1-14.2004
  5. Cindy D, AV Essen-Zandbergen, K Veldman, H Smit and D Mevius. 2010. Increased detection of extended spectrum beta-lactamase producing Salmonella enterica and Escherichia coli isolates from poultry. Vet. Microbiol. 145:273-278. https://doi.org/10.1016/j.vetmic.2010.03.019
  6. Clermont O, H Dhanji, M Upton, T Gibreel, A Fox, D Boyd, MR Mulvey, P Nordmann, E Ruppe, JL Sarthou, T Frank, S Vimont, G Arlet, C Branger, N Woodford and E. Denamur (2009). Rapid detection of the O25b-ST131 clone of Escherichia coli encompassing the CTX-M-15-producing strains. Journal of Antimicrobial Chemotherapy 64:274-277. https://doi.org/10.1093/jac/dkp194
  7. Clinical Laboratory Standard Institute. 2009. Performance standards for antimicrobial susceptibility testing: 9th informational supplement. M100-S19. Wayne. PA.
  8. Corvec S, L Cremet, C Leprince, S Dauvergne, A Reynaud, D Lepelletier and N Caroff. 2010. Epidemiology of Escherichia coli clinical isolates producing AmpC plasmidic $\beta$-lactamase during a 5-year period in a French teaching Hospital. Diagn. Microbiol. Infect. Dis. 67:277-281. https://doi.org/10.1016/j.diagmicrobio.2010.02.007
  9. Dierikx C, A van Essen-Zandbergen, K Veldman, H Smith and D Mevius. 2010. Increased detection of extended spectrum beta-lactamase producing Salmonella enteric and Escherichia coli isolates from poultry. Vet. Microbiol. 145:273-278. https://doi.org/10.1016/j.vetmic.2010.03.019
  10. Eckert C, V Gautier, M Saladin-Allard, N Hidri, C Verdet and Z Ould-Hocine. 2004. Dissemination of CTX-M-type $\beta$-lacta-mases among clinical isolates of Enterobacteriaceae in Paris, France. Antimicrob. Agents Chemother. 48:1249-1255. https://doi.org/10.1128/AAC.48.4.1249-1255.2004
  11. Edelstein M, M Pimkin, I Palagin, I Edelstein and L Stratchounski. 2003. Prevalence and molecular epidemiology of CTX-M extended-spectrum $\beta$-lactamase-producing Escherichia coli and Klebsiella pneumoniae in russian hospitals. Antimicrob. Agents Chemother. 47:3724-3732. https://doi.org/10.1128/AAC.47.12.3724-3732.2003
  12. Gona F, ML Mezzatesta, D Corona, D Zerobo, V Scriffignano, S Stefani, P Veroux and M Veroux. 2011. Klebsiella pneumonia ESBL producers responsible for severe UTIs in a renal transplant unit. Infection. 39:83-85. https://doi.org/10.1007/s15010-011-0081-0
  13. Horii T, K Tamai, M Mitsui, S Notake and H Yanagisawa. 2011. Blood stream infections caused by Acinetobacter ursingii in an obstetrics ward. Infect. Genet. Evol. 11:52-56. https://doi.org/10.1016/j.meegid.2010.10.011
  14. Hsieh CJ, YH Shen and KP Hwang. 2010. Clinical implications, risk factors and mortality following community-onset bacteremia caused by extended-spectrum $\beta$-lactamase (ESBL) and non-ESBL producing Escherichia coli. J. Microbiol. Immunol. Infect. 43:240-248. https://doi.org/10.1016/S1684-1182(10)60038-2
  15. Johann D, D Pitout, A Hossain and ND Hanson. 2004. Phenotypic and Molecular Detection of CTX-M-$\beta$-Lactamases Produced by Escherichia coli and Klebsiella spp. J. Clin. Microbiol. 42:5715-5721. https://doi.org/10.1128/JCM.42.12.5715-5721.2004
  16. Jung YS, HJ Hong, HR Nam and YH Lee. 2002. Isolation of norfloxacin resistant Escherichia coli from the han river and characterization of resistant mechanism. J. Microbiol. 40:63-69.
  17. Kim CK, JH Yum, DE Yong, SH Jeong, KW Lee and YS Chong. 2008. Detection of CTX-M-Type Extended-Spectrum $\beta$-lactamase in Clinical Isolates of Chromosomal AmpC $\beta$-Lactamase-Producing Enterobacteriaceae from Korea and Their Molecular Characteristics. Korean J. Clin Microbiol. 11:90-97. https://doi.org/10.5145/KJCM.2008.11.2.90
  18. Kim JM, YM Lim, I Rheem, YH Lee, JC Lee, SY Seol, YC Lee and DT Cho. 2005. CTX-M and SHV-12 $\beta$-lactamases are the most common extended-spectrum enzymes in clinical isolates of Escherichia coli and Klebsiella pneumoniae collected from 3 university hospitals within Korea. FEMS Microbiol. Lett. 245:93-98. https://doi.org/10.1016/j.femsle.2005.02.029
  19. Kim YM, HY Kang and YH Lee. 2008. The Identification of CTX-M-14, TEM-52, and CMY-1 Enzymes in Escherichia coli Isolated from the Han River in Korea. J. Microbiol. 46:478-481. https://doi.org/10.1007/s12275-008-0150-y
  20. Knothe H, P Shah, V Kremery, M Antal and S Mistuhashi. 1983. Transferable resistant to cefotaxime, cefoxitin, cefamandole, and cefuroxime in clinical isolates of Klebsiella pneumoniae and Serratia marcescens. Infection 11:315-317. https://doi.org/10.1007/BF01641355
  21. Lau SH, S Reddy, J Cheesbrough, FJ Bolton, G Willshw, T Cheasty, AJ Fox and M Upton. 2008. Major Uropahogenic Escherichia coli Strain Isolated in the Northwest of England Identified by Multilocus Sequence Typing. J. Clin. Microbiol. 46:1076-1080. https://doi.org/10.1128/JCM.02065-07
  22. Lau SH, ME Kaufmann, DM Livermore, N Woodford, GA Willshaw, T Cheasty, K Stamper, S Reddy, J Cheesbrough, FJ Bolton, AJ Fox and M Upton. 2008. UK epidemic Escherichia coli strains A-E, with CTX-M-15 $\beta$-lactamase, all belong to the international O25:H4-ST131 clone. J. Antimicrob. Chemother. 62:1241-1244. https://doi.org/10.1093/jac/dkn380
  23. Lee SH, MN Kim, SJ Choi and WS Chung. 2000. Characteristics of Extended-spectrum $\beta$-lactamase of Escherichia coli Strains Isolated from Clinical Specimens. Korean J. Clin. Pathol. 20:400-409.
  24. Lee SG, SH Jeong, HK Lee, CK Kim, YS Lee, EM Koh, YS Chong and KW Lee. 2009. Spread of CTX-M-type extended-spectrum $\beta$-lactamases among bloodstream isolates of Escherichia coli and Klebsiella pneumoniae from a Korean hospital. Diagn. Microbiol. Infect. Dis. 63:76-80. https://doi.org/10.1016/j.diagmicrobio.2008.09.002
  25. Lewis II JS, M Herrera, B Wickes, JE Patterson and JH Jorgensen. 2007. First report of the emergence of CTX-M-type extended-spectrum $\beta$-lactamases (ESBLs) as the predominant ESBL isolated in a U.S. health care system. Antimicrob. Agents Chemother. 51:4015-4021. https://doi.org/10.1128/AAC.00576-07
  26. Mora A, A Herrera, R Mamani, C Lopez, MP Alonso, JE Blanco, M Blanco, G Dahbi, F Garcia-Garrote, JM Pita, A Coira, MI Bernardez and J Blanco. 2010. Recent emergence of clonal group O25b:K1:H4-B2-ST131 ibeA strains among Escherichia coli poultry isolates, including CTX-M-9-producing strains, and comparison with clinical human isolates. Appl. Environ. Microbiol. 76:6991-6997. https://doi.org/10.1128/AEM.01112-10
  27. Oh CE, JS Hong, IK Bae, EH Song, SH Jeong and KW Lee. 2005. Dissemination of CTX-M type extended-spectrum $\beta$-lactamases and emergence of CTX-M-12 in Escherichia coli. Korean J. Lab Med. 25:252-258.
  28. Oppegaard H, TM Steinum and Y Wasteson. 2001. Horizontal transfer of a multi-drug resistant plasmid between coli form bacteria of human and bovine origin in a farm environment. Appl. Environ. Microbiol. 67:3732-3734. https://doi.org/10.1128/AEM.67.8.3732-3734.2001
  29. Paterson DL and RA Bonomo. 2005. Extended-Spectrum $\beta$-lactamase: a clinical update. Clin. Microbiol. Rev. 18:657-686. https://doi.org/10.1128/CMR.18.4.657-686.2005
  30. Perrentin V, F Schwarz, L Cresta, M Boeglin, G Dasen and M Tenber. 1997. Antibiotic resistant spread in food. Nature. 389:801-802.
  31. Peirano G, CHJ Van Grune and JDD Pitout. 2011. Characteristics of infections caused by extended-spectrum $\beta$-lactamaseproducing Escherichia coli from community hospitals in South Africa. Diagn. Microbiol. Infect. Dis. 69:449-453. https://doi.org/10.1016/j.diagmicrobio.2010.11.011
  32. Peirano G and JD Pitout. 2010. Molecular epidemiology of Escherichia coli producing CTX-M $\beta$-lactamase: the worldwide emergence of clone ST131 O25:H4. Int. J. Antimicrob. Agents. 35:316-321. https://doi.org/10.1016/j.ijantimicag.2009.11.003
  33. Pitout JD and KB Laupland. 2008. Extended-spectrum betalactamase-producing Enterobacteriaceae: and emerging public-health concern. Lancet Infect Dis. 8:159-166. https://doi.org/10.1016/S1473-3099(08)70041-0
  34. Philippon A, G Arlet and PH Lagrange. 1994. Origin and impact of plasmid-mediated extended spectrum beta-lactamases. Eur. J. Clin. Microbiol. Infect. Dis. 13:17-29. https://doi.org/10.1007/BF02390681
  35. Poirel L, M Gniadkowski, P Nordmann. 2002. Biochemical analysis of the ceftazidime-hydrolysing extended-spectrum $\beta$-lactamase CTX-M-15 and of its structurally related $\beta$-lactamase CTX-M-3. J. Antimicrob. Chemother. 50:1031-1034. https://doi.org/10.1093/jac/dkf240
  36. Qi C, V Pilla, JH Yu and K Reed. 2010. Changing prevalence of Escherichia coli with CTX-M-type extended-spectrum $\beta$-lactamases in outpatient urinary E. coli between 2003 and 2008. Diagn. Microbiol. Infect. Dis. 67:87-91. https://doi.org/10.1016/j.diagmicrobio.2009.12.011
  37. Qureshi AR and MA Qureshi. 1992. Multiple antibiotic resistant feacal coliform in raw sewage. Water Air Soil Pollut. 61:47-56. https://doi.org/10.1007/BF00478365
  38. Reinthaler FF, G Feierl, H Galler, D Haas, E Leitner, F Mascher, A Melkes, J Posch, I Winter, G Zarfel and E Marth. 2010. ESBL-producing E. coli in Austrian sewage sludge. Water Res. 44:1981-1985. https://doi.org/10.1016/j.watres.2009.11.052
  39. Rossolini GM, MM D'andrea and C Mugnailoi. 2008. The spread of CTX-M-type extended-spectrum $\beta$-lactamases. Clin. Microbiol. Infect. 14:33-41. https://doi.org/10.1111/j.1469-0691.2007.01867.x
  40. Smalla K and PA Sobecky. 2002. The prevalence and diversity of mobile genetic elements in bacterial communities of different environmental habitats: insights gained from different methodological approaches. FEMS Microbiol. Ecol. 42:165-175. https://doi.org/10.1111/j.1574-6941.2002.tb01006.x
  41. Smith P, M Hiney and O Samuelson. 1994. Bacterial resistant to antimicrobial agents used in fish farming; a critical evaluation of method and meaning. Annu. Rev. Fish Dis. 4:273-313.
  42. Tenover FC, PM Raney, PP Williams, JK Rasheed, JW Biddle, A Oliver, SK Fridkin, L Jevitt and JE McGowan Jr. 2003. Evaluation of the NCCLS extended-spectrum beta-lactamase confirmation methods for Escherichia coli with isolates collected during Project ICARE. J. Clin. Microbiol. 41:3142-3146. https://doi.org/10.1128/JCM.41.7.3142-3146.2003
  43. Thomas T, R Szczepanowski, S Braun, A PUhler SchlUter. 2003. Occurrence of integron-associated resistant gene cassettes located on antibiotic resistant plasmids isolated from a wastewater treatment plant. FEMS Microbiol. Ecol. 45:239-252. https://doi.org/10.1016/S0168-6496(03)00164-8
  44. Tomanicek SJ, KK Wang, KL Weiss, MP Blakeley, J Cooper, Y Chen and L Coates. 2010. The active site protonation states of perdeuterated Toho-1 $\beta$-lactamase determined by neutron diffraction support a role for Glu166 as the general base in acylation. FEBS Lett. 585:364-368.
  45. Walther-Rasmussen J and N HOiby. 2004. Cefotaximase (CTXM-ases), an expanding family of extended-spectrum $\beta$-lactamase. Can. J. Microbiol. 50:137-165. https://doi.org/10.1139/w03-111
  46. Wannaprasat W, P Padungtod and R Chuanchuen. 2011. Class 1 integrons and virulence genes in Salmonella enterica isolates from pork and humans. Int. J. Antimicrob. Agents. 37:457-461. https://doi.org/10.1016/j.ijantimicag.2010.12.001
  47. Wirth T, D Falush, R Lan, F Colles, P Mensa, LH Wieler, H Karch, PR Reeves, MCJ Maiden, H Ochman and M Achtman. 2006. Sex and virulence in Escherichia coli: an evolutionary perspective. Mol. Microbiol. 60:1136-1151. https://doi.org/10.1111/j.1365-2958.2006.05172.x
  48. Zou LK, HN Wang, B Zeng, AY Zhang, JN Li, XT Li, GB Tian, K Wei, YS Zhou, CW Xu and ZR Yang. 2011. Phenotype and genotypic characterization of $\beta$-lactam resistant in Klebsiella pneumoniae isolated from swine. Vet. Microbiol. 149:139-146. https://doi.org/10.1016/j.vetmic.2010.09.030