Isolation, characterization and molecular evaluation of genetic factors of antibiotic resistance in pathogenic Escherichia coli

Document Type : Full Research Paper

Author

Department of Microbiology, Fasa University of Medical Sciences, Fasa, Iran & Deprtment of Biology, Payame Noor Unvierstiy (PNU), Tehran, Iran.

Abstract

The usage of antimicrobial drugs in the veterinary field can cause the emergence, prevalence, and dissemination of antimicrobial resistance in bacteria isolated from food-producing animals. Antimicrobial drugs are essential for the treatment of bacterial infections in cattle. Class 1 integrons can extend antibiotic resistance genes among bacteria Escherichia coli from animal.The objective of this study was to investigate the antimicrobial resistance pattern and the frequency class 1 integrons Escherichia coli strains isolated from diarrheic calves in Fars province. A one hundred and twenty samples from 120 different diarrheic calves from Fars province in Iran were screened for the presence of virulence factors associated with diarrhea. Following isolation and identification of Escherichia coli strains on the basis of cultural, morphological and biochemical properties, the presence of various virulence genes of ETEC (k99, F41 fimbria and Sta), EHEC (Stx1, Stx2 and eae) was investigated by multiplex PCR. The isolates were examined for susceptibility to antimicrobial agent and class 1 integrons using PCR and agar disk-diffusion method for 8 antimicrobial agents. Out of 120 samples, 57 (47/5%) were found to be positive for Escherichia coli and 35 of them identified as pathogenic Escherichia coli including 21( 36/8%) and 14 (24/5%) ETEC and EHEC strains respectively. The most antibiotic resistance level was seen against erythromycin (100%), penicillin (100%) co-trimoxazole (100%), tetracycline (91/5%), ampicillin (88/5%) and chloramphenicol (83%). All isolates shown MDR . The study of molecular results showed that class 1 integron was detected in (48/75%) of isolates. Our data demonstrate the presence of MDR and class 1 integrons were common in Escherichia coli from calves in Fars province experiencing problems with neonatal diarrhea.

Keywords

Main Subjects

References

1. Aarestrup, F. M. 1999. Association between the consumption of antimicrobial agents in animal husbandry and the occurrence of resistant bacteria among food animals. International Journal of Antimicrobial Agents. 12 (4):279–285.
2. Abdelhaleem, A. A., Homeda, H. E., Hershan, A. A., Makeen, A. M., Alsanosy, R. M. 2014. Class 1 Integrons Gene in Drug Resistant E. coli and Isolated from Different Clinical Specimens in Jazan Area K.S.A. International Journal of Current Research. 6(1): 9283-9286.
3. Afshar Mazndaran, N., Rajab, A. 2011. Correct prescription of antibiotics in the treatment of diseases of poultry. 2nd ed. Tehran: Press Noorbakhsh.P.184-185. (Book In Farsi)
4. Arjmandpour, P., Tahamtan, Y. 2017. Molecular Identification of Virulence Factors of E. coli in Three Strains of ETEC, EHEC and EPEC in Calves and Lambs with Diarrhea in Shiraz. Veterinary Journal (Pajouhesh & Sazandegi), 115:27-34. (In Persian)
5. Constable, P. D., 2004. Antimicrobial use in the treatment of calf diarrhea. Journal of veterinary internal medicine, 18(1), 8-17.‏
6. Du, X., Shen, Z., Wu, B., Xia, S., & Shen, J. (2005). Characterization of class 1 integrons-mediated antibiotic resistance among calf pathogenic Escherichia coli. FEMS microbiology letters, 245(2), 295-298.‏
7. El-Sokkary, M. A., Abdelmegeed, E. S. 2015. Characterisation of Class 1 Integron among Escherichia coli Isolated from Mansoura University Hospitals in Egypt. Advances in Microbiology. (1): 269-277.
8. Grape, M., Farra. A., Kronvall. G., Sundstrom. L. 2005. Integrons and gene cassettes in clinical isolates of co-trimoxazole-resistant gram-negative bacteria. Journal of Clinical Microbiology.11(1):185-92.
9. Harada, K., Asai, T. 2010. Role of Antimicrobial Selective Pressure and Secondary Factors on Antimicrobial Resistance Prevalence in Escherichia coli from Food-Producing Animals in Japan. Journal of Biomedicine and Biotechnology. 10(1):1-12.
10. Hasani Tabatabai. A., Firozi. R. 2011. University of Tehran Press (UTP). (Book in Persian)
11. Islami, G., Seyedjavadi, S., Goudarzi, H., Fallah, F., Goudarzi, M .2010. The prevalence integrons of in Escherichia coli Klebsiella in children with urinary tract infection with multi-drugresistant. Research Journal of Medical Sciences Faculty and Health Services of Shahid Beheshti Tehran.34(1):61-65. ( In Farsi)
12. Jones, L. A., McIver, C. J., Kim, M. J., Rawlinson, W. D., White, P.A. 2005. The aadB gene cassette is associated with blaSHV genes in Klebsiella species producing extended-spectrum beta-lactamases. Antimicrob Agents Chemother. 49(2): 794-797.
13. Kadlec, K., Schwarz, S .2008. Analysis and distribution of class 1 and class 2 integrons and associated gene cassettes among Escherichia coli isolates from swine, horses, cats and dogs collected in the BfT-GermVet monitoring study. Journal of Antimicrobial Chemotherapy. 62(3):469-473.
14. Kang, H.Y., Jeong, J.S., Oh, J.Y., Tae, S.H., Choi, C. H., Moon, D.C. Lee, W.K., Lee, Y.C., Seol, S.Y., Cho, D.T., Lee, J.C. 2005. Characterization of antimicrobial resistance and class 1 integrons found in Escherichia coli isolates from humans and animals in Korea. Journal of Antimicrobial Chemotherapy. 55(1): 639–644.
15. Levin, B.R., Lipsitch, M., Perrot, V., Schrag, S., Antia, R., Simonsen, L., Moore Walker, N., Stewart, F. M. 1997. The population genetics of antibiotic resistance. Clinical Infectious Diseases.(Suppl 1):S9-16.
16. Rao, A. N., M. Barlow, L. A. Clark, J. R. Boring, F. C. Tenover and J. J. E. McGowan, 2006. Class 1 Integrons in Resistant Escherichia coli and Klebsiella spp., US Hospitals. Emerging Infectious Diseases. 12: 1011-1014.
17. Sarshar, M., Tavoffi, H., Ghorbani Dalin, S., Saudi,N., Kargar, M., Shahrokh, N.2012. Antibiotic resistance and distribution of tetracycline resistance genes in Escherichia coli strains producing diarrhea isolated from children. Iranian Journal of Infectious Diseases and Tropical Medicine, 57(17):131-136. (In Persian)
18. Severino, P., Magalhaes, V.D. 2004. Integrons as tools for epidemiological studies. Journal of Antimicrobial Chemotherapy. 10(2): 156-162.
19. Singh, R., Schroeder, C. M., Meng, J., White, D. G., McDermott, R. F., Wagner, B. D.D., Yang, H., Simjee, S., Debroy, C., Walker, R.D., Zhao, S. 2005. Identification of antimicrobial resistance and class 1 integrons in Shiga toxin-producing Escherichia coli recovered from humans and food animals. Journal of Antimicrobial Chemotherapy. 56(1):216-219.
20. Tzipori, S. 1985. The relative importance of enteric pathogens affecting neonates of domestic animals. Advances in Veterinary Science and Comparative Medicine. 29(1): 103-206.
21. Verdier, K.D., Nyman, A., Greko, C., Bengtsson, B. 2012. Antimicrobial resistance and virulence factors in Escherichia coli from Swedish dairy calves. Acta Veterinaria Scandinavica. 54(1):2-10.
22. White, P. A., Mciver, C.H. J., Rawlinson, W. 2001. Integrons and Gene Cassettes in the Enterobacteriaceae. Antimicrob Agents Chemother. 45 (9): 2658–2661.
23. Yaghubzadeh, N,. Onagh, A., Mardani, K,. Khalili, M., Tukmuchi, A,. Nick Bakhsh, P,. 2011. molecular characterization, Study of Antibiotic Resistance Pattern of Escherichia coli Producing Shiga Toxin and Antibacterial Effects of Thyme and Cumin Essences Against them. Urmia Medical Journal, NO: 3. (In Persian)
24. Yu, H.S., Lee, J.C., Kang, H.Y., Ro, D.W., Chung, J.Y., Jeong, Y.S., Tae, S. H., Choi, C. H., Lee, E. Y., Seol, S.Y., Lee ,Y. C., Cho, D.T .2003. Changes in gene cassettes of class 1 integrons among Escherichia coli isolates from urine specimens collected in Korea during the last two decades. Journal of Clinical Microbiology. 41(12): 5429-5433. 

Files

History

  • Receive Date: 03 July 2017
  • Revise Date: 07 March 2018
  • Accept Date: 07 March 2018

Share

How to cite

Statistics