Identification and characterization of shiga toxin-producing Escherichia coli isolated from the feces of slaughtered pigs

도축돈 장분변으로부터 Shiga Toxin-Producing Escherchia coli의 분리 와 성상

  • Song, Young-hwan (College of Veterinary Medicine, Chungnam National University) ;
  • Kim, Ji-young (College of Veterinary Medicine, Chungnam National University) ;
  • Chae, Mi-kyung (College of Veterinary Medicine, Chungnam National University) ;
  • Park, Chang-sik (Research Center for Transgenic Cloned Pigs, Chungnam National University) ;
  • Kim, Myung-chul (College of Veterinary Medicine, Chungnam National University) ;
  • Jun, Moo-hyung (College of Veterinary Medicine, Chungnam National University)
  • 송영환 (충남대학교 수의과대학) ;
  • 김지영 (충남대학교 수의과대학) ;
  • 채미경 (충남대학교 수의과대학) ;
  • 박창식 (충남대학교 형질전환복제돼지연구센터) ;
  • 김명철 (충남대학교 수의과대학) ;
  • 전무형 (충남대학교 수의과대학)
  • Accepted : 2004.10.05
  • Published : 2004.12.30

Abstract

Shiga toxin-producing Escherichia coli (STEC) causes various clinical signs in human and animals, and has been indicated as a global enteropathogen with zoonotic importance. In this study, the feces of healthy pigs were collected from the slaughtered pigs of Daejon abattoir during the period from December 2001 to October 2002. Of 326 specimens, 13 STEC were confirmed by culture, PCR and colony hybridization. The isolates were further studied for toxin types, pathogenic factors, plasmid profiles, and antimicrobial resistance to characterize the genetic and toxigenic properties. In PCR, all of 13 isolates were evident to have shiga toxin gene (stx). Of 13 isolates stx1 gene was detected in 4 and stx2 gene in 9. The genes of eaeA, hlyA and rfbE were not present in any isolates. In colony hybridization using shiga toxin common primer (STXc), 2 to 9 per 100 colonies subcultured from 13 isolates showed the positive reaction. In the examination for plasmid profiles of the isolates, one to eleven plasmids with varying sizes of 1.0 Kb to 100 Kb were detected, and the 13 STEC could be classified into four groups by the plasmid patterns. The antimicrobial resistance patterns of the isolates were comparably corresponded with the plasmid profile patterns.

Keywords

References

  1. 고주언, 홍종해. 도체 표면의 분변오염과 verotoxin 생성 Escherichia coli O157:H7 분리에 관한 연구. 한국식품위생안전성학회지. 1997, 12, 78-82
  2. 박주연, 신나리, 박용호, 유한상. 설사 자돈으로부터 분리한 Escherichia coli의 특성에 관한연구; 항균제 감수성, 장독소 및 섬모의 유전형의 분포 및 plasmid profile. 대한수의학회지. 2000, 40, 301-310
  3. 정석찬, 정병열, 윤장원, 조윤상, 김종염, 박용호. 쇠고기 중 Escherichia coli O157:H7 신속검출을 위한 multiplex-PCR기법 개발. 대한수의학회지. 1998, 38, 173-181
  4. 정수관, 정석찬, 최원필. 돼지 유래 대장균의 생물학적 특성과 plasmid profile 에 대하여. 대한수의학회지. 1990,30, 287-295
  5. 진형근, 차인호, 김용환. 설사환자로부터 verotoxin 생산대장균의 분리 및 생물 학적 독성. 한국수의공중보건학회지. 1996, 20, 89-96
  6. 차인호, 김용환. 동물분변에서 Escherichia coli O1557:H7의 분리 및 이들 균이 생산하는 verotoxin-2의 생물화학적 특성. 대한수의학회지. 1996, 36, 371-378
  7. Beutin, L. Geier, D. Steinruck, H., Zimmermann, S. and Scheuts, F. Prevalence and some properties of verotoxin(Shiga-like toxin)-producing Escherichia coli in seven different species of healthy domestic animals. J. Clin. Microbiol. 1993, 31, 2483-2488
  8. Botteldoorn, N., Heyndrickx, M., Rijpens, N. and Herman, L. Detection and characterization of verotoxigenic Escherichia coli by a VTEC/EHEC multiplex PCR in porcine faeces and pig carcass swabs. Res. Microbiol. 2003, 154, 97-104
  9. Bulte, M., Montenegro, M. A., Helmuth, R., Trumpf, T. and Reuter, G. Detection of verotoxin-producing E. coli(VTEC) in healthy cattle and swine with DNADNA colony hybridization method. Berl. Munch. Tierarztl. Wochenschr. 1990, 103, 380-384
  10. Clarke, S. C., Haigh, R. D., Freestone, P. P. and Williams, P. H. Virulence of enteropathogenic Escherichia coli, a global pathogen. Clin. Microbiol. Rev. 2003, 16, 365-378
  11. Farmer, J. J. 3rd and Davis, B. R. H7 antiserumsorbitol fermentation medium: a single tube screening medium for detecting Escherichia coli O157:H7 associated with hemorrhagic colitis. J. Clin. Microbiol. 1985, 22, 620-625
  12. Fukushima, H., Hoshina, K. and Gomyoda, M. Long-term survival of shiga toxin-producing Escherichia coli O26, O222, and O157 in bovine feces. App. Environ. Microbiol. 1999, 65, 5177-5181
  13. Gannon, V. P. J., D’souza, S., Graham, T., King, R. K., Rahn, K. and Read, S. Use of the flagella H7 gene as a target in multiplex PCR assays and improved specificity in identification of enterohemorrhagic Escherichia coli strains. J. Clin. Microbiol. 1997, 35, 656-662
  14. Gannon, V. P., Gyles, C. L. and Friendship, R. W. Characteristics of verotoxigenic Escherichia coli from pigs. Can. J. Vet. Res. 1998, 52, 331-337
  15. Herbert, S., Lothar, B. and Helge, K. Molecular analysis of the plasmid-encoded hemolysin of Escherichia coli O157:H7 strain EDL 933. Infect. Immun. 1995, 63, 1055-1061
  16. Hiroshi, A., Sou-ichi, M., Toshikazu, S., Teizo, T., Hisao, K., Tetsuya, I. and Kouichi, T. Detection and long-term existence of shiga toxin(Stx)-producing Escherichia coli in sheep. Microbiol. Immunol. 1998, 42, 683-688
  17. James, P. N. and Kaper, J. B. Diarrheagenic Escherichia coli. Clin. Microbiol. Rewiews. 1998, 11, 142-201
  18. Kaper, J. B., Nataro, J. P. and Mobley, H. L. Pathogenic Escherichia coli. Nat. Rev. Microbiol. 2004, 2, 123-140
  19. Maite, M. and Juan, J. Abundance in sewage of bacteriophages that infect Escherichia coli O157:H7 and that carry the shiga toxin 2 gene. Appl. Environ. Microbiol. 1998, 64, 2443-2448
  20. Masahisa, W., Toshio, S., Midori, K., Takeshi, S., Shinji, Y., Toru, T., Chihiro, S. and Yoshifumi, T. Identification and characterization of a newly isolated shiga toxin 2-converting phage from shiga toxinproducing Escherichia coli. Infect. Immun. 1998, 66, 4100-4107
  21. Osek, J. Prevalence of virulence factors of Escherichia coli strains isolated from diarrhegic and healthy piglets after weaning. Vet. Microbiol. 1999, 68, 209-217
  22. Phillip, I. T., Laura, M. S., Yoo-lee, Y., Teresa, R. W., Srdjan, J. and Thomas, S. W. Acquisition of the rfb-gnd Cluster in Evolution of Escherichia coli O55 and O157. J. Bacteriol. 2000, 182, 6183-6191
  23. Rowland, C. and Patricia, D. Characterisation and clonal relationships of Shiga-toxigenic Escherichia coli(STEC) isolated from Austraila dairy cattle. Vet. Microbiol. 2001, 79, 323-335
  24. Sambrook, J. and Russell, D. W. Molecular cloning a laboratory manual. 3rd ed, Cold Spring Harbor Press, New York, 2001
  25. Terezinha, K., Maria, R. B., Andrea, M. M., Tania, A. T. G., Monica, A. M. V., Claudete, S. A. F. and Antonio, J. P. F. Virulence properties of Escherichia coli isolated from ostriches with respiratory disease. Vet. Microbiol. 2001, 83, 71-80
  26. Tsukamoto, T. and Kawai, T. Identification of Escherichia coli O157 antigen by polymerase chain reaction. Kansenshogaku Zasshi. 1998, 72, 738-741