Detection of Shiga-Toxin Producing E.coli (STEC), Enteropathogenic E.coli (EPEC) And Enterotoxigenic E.coli (ETEC) From Animals By M-PCR

Abstract

This research investigated the presence of virulence genes encoding F41, K99, eae, Stx1, Stx2 and STa and the antimicrobial resistance of animal Escherichia coli (E. coli) isolates. Clinical isolates (n:233) were evaluated from faecal samples of cattle, sheep, goats, horses, cats and dogs collected between the years of 2010 to 2015 from Turkey. Enterohaemorrahic E.coli (EHEC) O157:H7 was detected by using cefixime tellurite sorbitol MacConkey agar (CT-SMAC) and Wellcollex E. coli (Remel®). The Kirby-Bauer disc diffusion test was performed to detect the resistance pattern of the isolates to ampicillin, Amoxycillin/clavulanic acid, enrofloxacin, ceftiofur, trimethoprim/sulfamethoxazole and tetracycline. The results showed that 40% of the ruminant isolates were identified as Shiga-toxin producing Escherichia coli (STEC).  Enterotoxigenic E. coli (ETEC) was detected in samples from cattle (0.9%) and sheep (12%). Enteropathogenic E.coli (EPEC) was detected in samples from cattle (0.9%) and dogs (11.4%). EHEC O157:H7 was not detected any of the isolates. Among all E. coli isolates that carried at least one virulence gene, 8 (19%) were resistant to more than three antimicrobials, 7 (16.7%) were resistant to at least one antimicrobial and 27 (64.3%) were susceptible to all antimicrobials.

Keywords

• Antimicrobial susceptibility,E. coli,Pcr,EHEC,EPEC,ETEC,STEC

References

1. 1. Jorgensen JH, Pfaller MA, Carroll K C, Funke G, Landry ML, Richter SS, Warnock DW (2015):Manual of Clinical Microbiology. Volume 1.11th edition, ASM Press, Washington, DC, pp: 685-713.Chapter 37. Esherichia, Shigella and Salmonella. Strockbine N.A.., Bopp C.A., Fields P.I., Kaper J.B., Nataro J.P.
2. 2. Gyles CL (2007): Shiga toxin-producing Escherichia coli: an overview. Journal of Animal Science 85, 45-62.
3. 3. Nataro JP, Kaper JB (1998): Diarrheagenic Escherichia coli. Clinical Microbiology Reviews 11,142-201.
4. 4. Gyles CL, Fairbrother JM (2010): Escherichia coli, in: Gyles, C. L., Prescott, J. F., Songer, J. G. and Thoen, C. O. (eds), Pathogenesis of Bacterial Infections in Animals.4th ed.Wiley-Blackwell, Ames, IA, U.S.A, 267-308.
5. 5. Kaper JB, Nataro JP, Mobley HL (2004): Pathogenic Escherichia coli. Nature Reviews Microbiology 2, 123-140.
6. 6. Law D (2000): Virulence factors of Escherichia coli O157 and other shiga toxin-producing E.coli. Journal of Applied Microbiology 88, 729-745.
7. 7. Blanco M, Blanco J, Blanco JE, Ramos J (1993): Enterotoxigenic, verotoxigenic, and necrotoxigenic Escherichia coli isolated from cattle in Spain. Am. J. Vet. Res. 54, 1446-1451.
8. 8. Pitout, JD, Laupland KB (2008): Extended-spectrum beta-lactamase-producing Enterobacteriaceae: an emerging public health concern. Lancet Infectious Disease 8, 159-166.

9. 9. Sawant AA, Hegde NV, Straley BA, Donaldson SC, Love BC, Knabel SJ, Jayarao BM (2007): Antimicrobial-Resistant Enteric Bacteria from Dairy Cattle. Applied and Environmental Microbiology 73, 156-163.
10. 10. Franck SM, Bosworth BT, Moon HW (1998): Multiplex PCR for enterotoxigenic, attaching and effacing, and Shiga toxin- producing Escherichia coli strains from calves. Journal of Clinical Microbiology 36, 1795-1797.
11. 11. Anonymous: European Committee on Antimicrobial Susceptibility Testing Breakpoint tables for interpretation of MICs and zone diameters Version 7.1, valid from 2017-03-10 http://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/Breakpoint_tables/v_7.1_Breakpoint_Tables.pdf.(12.19.2017)
12. 12. Guler L, Gunduz K, Ok U (2008): Virulence Factors and Antimicrobial Susceptibility of Escherichia coli Isolated from Calves in Turkey. Zoonoses and Public Health. 55, 249-257.
13. 13. Galli L, Miliwebsky E., Irino K, Leotta G, Rivas M (2010): Virulence profile comparison between LEE-negative Shiga toxin-producing Escherichia coli (STEC) strains isolated from cattle and humans. Veterinary Microbiology 143, 307-313.
14. 14. Nguyen TD, Vo TT, Vu-Khac H (2011): Virulence factors in Escherichia coli isolated from calves with diarrhea in Vietnam. Journal of Veterinary Science 12, 159-164.
15. 15. Barett T J, Kaper JB, Jerse A E, Wilson JB, Johnson RP, Gyles, CL (1992):Virulence factors in shiga-like toxin producing E. coli isolated from humans and cattle. Journal of Infectious Disesase 165, 979-980.
16. 16. Beutin L, Geier D, Steinruck H, Zimmermann S, Scheutz, F. (1993): Prevalence and Some Properties of Verotoxin (Shiga-Like Toxin)-Producing Escherichia coli in Seven Different Species of Healthy Domestic Animals. Journal of Clinical Microbiology 31, 2483-2488.
17. 17. Leomil L, Aidar-Ugrinovich L, Guth BE, Irino K, Vettorato M P, Onuma DL, de CastroA F. (2003): Frequency of Shiga toxin-producing Escherichia coli (STEC) isolates among diarrheic and non-diarrheic calves in Brazil. Veterinary Microbiology 97,103-109.
18. 18. Blanco J, Gonzalez EA, Garcia S (1988): Production of toxins by Escherichia coli strains isolated from calves with diarrhea in Galicia (north-western Spain). Veterinary Microbiology 18, 297-311.
19. 19. Turkyilmaz S, Eskiizmirliler S, Tunaligil S, Bozdogan B (2013): Identification, characterization and molecular epidemiology of Escherichia coli isolated from lamb and goat kids with diarrhea. Acta Veterinaria Brno 82, 357-362.
20. 20. De Verdier K, Nyman A, Greko C, Bengtsson B (2012): Antimicrobial resistance and virulence factors in Escherichia coli from Swedish dairy calves. Acta Veterinaria Scandinavica 54,2.
21. 21. Holland R E, Wilson RA, Holland M S, Yuzbasiyan-Gurkanc V, Mullaney TP, White DG(1999): Characterization of eae+ Escherichia coli isolated from healthy and diarrhoeic calves. Veterinary Microbiology 66, 251-263.
22. 22. Khakhria R, Duck D, Lior H (1990): Extended phage typing scheme for Escherichia coli 0157:H7. Epidemiology & Infection 105, 511-520.
23. 23. Beutin L (1999): Escherichia coli as a pathogen in dogs and cats. Veterinary Research 30,285- 298.24. Puno-Sarmiento J, Medeiros L, Chiconi C, Martins F, Pelayo J, Rocha S, Blanco J, Blanco M, Zanutto M, Kobayashi R, Nakazato G (2013): Detection of diarrheagenic Escherichia coli strains isolated from dogs and cats in Brazil. Veterinary Microbiology 166, 676-680. 25. Bryan A, Shapir N, Sadowsk MJ (2004): Frequency and Distribution of Tetracycline resistance genes in genetically diverse, non selected, and non clinical Escherichia coli strains isolated from diverse human and animal sources. Applied and Environmental Microbiology 70, 2503-2507.26. Talukdar PK, Rahman M, Rahman M, Nabi A, Islam Z, Hoque M M, Endtz HP, Islam MA (2013): Antimicrobial resistance, virulence factors and genetic diversity of Escherichia coli isolates from household water supply in Dhaka, Bangladesh. PLoS ONE. 8, e61090.27. Jacoby GA, Archer GL (1991): New mechanisms of bacterial resistance to antimicrobial agents. The New England Journal of Medicine 324, 601- 612.
24. 24. Puno-Sarmiento J, Medeiros L, Chiconi C, Martins F, Pelayo J, Rocha S, Blanco J, Blanco M, Zanutto M, Kobayashi R, Nakazato G (2013): Detection of diarrheagenic Escherichia coli strains isolated from dogs and cats in Brazil. Veterinary Microbiology 166, 676-680.
25. 25. Bryan A, Shapir N, Sadowsk MJ (2004): Frequency and Distribution of Tetracycline resistance genes in genetically diverse, non selected, and non clinical Escherichia coli strains isolated from diverse human and animal sources. Applied and Environmental Microbiology 70, 2503-2507.
26. 26. Talukdar PK, Rahman M, Rahman M, Nabi A, Islam Z, Hoque M M, Endtz HP, Islam MA (2013): Antimicrobial resistance, virulence factors and genetic diversity of Escherichia coli isolates from household water supply in Dhaka, Bangladesh. PLoS ONE. 8, e61090.
27. 27. Jacoby GA, Archer GL (1991): New mechanisms of bacterial resistance to antimicrobial agents. The New England Journal of Medicine 324, 601- 612