Molecular analysis of antibiotic resistance genes in Escherichia coli from broiler chickens in shahrebabak by Multiplex PCR Technique

Document Type : Research Paper


1 Ph. D. Candidate, Department of Microbiology, Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Professor, Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

3 Professor, Department of Microbiology, Science and Research Branch, Islamic Azad University, Tehran, Iran


Avian pathogenic Escherichia coli (APEC) is responsible for an extremely wide range of extra intestinal diseases in poultry, including Colibacillosis and Cellulitis. There is the problem of antibiotic resistance all over the world. The aim of this study is the molecular study of antibiotic resistance genes, such as: aac, aad, qnr, tet, anr, sul and determination of antibiotic susceptibility. 83 strains of E. coli of Colibacillosis cases and 34 strains of Cellulitisin cases were gotten from in poultry slaughterhouses in Shahrebabak province within 6 months in 2017. Strains of E. coli were confirmed by biochemical methods. The results of the study showed that 63.85% for the tetA gene, 62.65% for the tetB gene were positive. The abundance of genes qnrs (49.3%), qnrB (24.09%), sul1 (28.91%), dhfrv (31.32%), dhfrA (26.50%), aac (3)-1(25.66%), aadA (22.89%), floR (28.91%), sul1 (33.13%), qnrA (12.30%). None of the specimens was positive for blaoxa, blaCTX, blasHV, blaTEM, genes. In this study, it was found that all of the suppositories were resist ant to one or more antibiotics between 40% and 90%. The highest resistance to antibiotics was tetracycline, sulfamethoxazole, chloramphenicol, terimetoprim. In this study, antibiotic resistance was found to be due to the presence of resistance genes. Recognition of resistance pattern and microorganisms susceptibility to antibiotics have an effective role on correct and suitable selection of antibiotic and infection control.


  1. Ahsani, M. R., Bafti, M. S., Esmailizadeh, A. K. & Mohammadabadi, M. R. (2011). Genotyping of isolates of Clostridium perfringens from vaccinated and unvaccinated sheep. Small Ruminant Research, 95(1), 65-69.
  2. Ahsani, M. R., Mohammadabadi, M. R. & Shamsaddini, M. B. (2010). Clostridium perfringens isolate typing by multiplex PCR. Journal of Venomous Animals and Toxins including Tropical Diseases, 16(4), 573-578.
  3. Barnes, H. J., Nolan, L. K. & Vailan Court, J. P. (2008). Colibacillosis. In: Saif Y. M., Fadly A. M., Glisson J.R., McDougald L.R., Nolan L.K., Swayne D.E.: Diseases of Poultry. 12th Ed., Blackwell publishing, chapter 18, 691-738.
  4. Dho, M. & Lafont, J. P. (2009). Adhesive properties and iron uptake abilities in E. coli lethal and non-lethal for chicks. Avian Diseases, 28, 1116-1125.
  5. 5-Escobar-Paramo, P., Clermont, O., Blanc-Potard, A. B., Bui, H., Bouguenec C. Le. & Denamur, E. (2004a). A specific genetic background is required for acquisition and expression of virulence factors in Escherichia coli. Mol Biol Evol, 21, 1085-1094.
  6. 6-Ghanbarpour, R. (2017). Genetipicanalysis of virulence genes antimicrobial profile of diarrrea agenic Escherichia coli isolaited from disease poultray in Iran tropanima healtprod. Dol 10. 1007/s 11250-017-1234-7.
  7. Gordon, D. M., Clermont, O., Tolley, H. & Denamur, E. (2011). Assigning Escherichia coli strains to phylogenetic groups: multi-locus sequence typing versus the PCR triplex method. Environ Microbiol, 10, 2484-2496.
  8. Fankhauser, C., Schrenzel, J., Prendki, V., Ris, F., Schiffer, E., Gastmeier, P. & Harbarth, S. (2015). Prevalence of extended-spectrum betalactamase producing–Enterobacteriaceae (ESBL-E) carriage on admission at Geneva University Hospitals (HUG). Antimicrob Resist Infect Control, 4(1), 120.
  9. Facklam, R. R., Carvalho, M. & Teixeira, L. M. (2012). History, taxonomy, biochemical characteristics, and antibiotic susceptibility testing of enterococci. Washington DC: ASM Science, 1-54.
  10. Hadizadeh, M. R., Mohammadabadi, A., Niazi, A. K., Esmailizadeh Koshkoiyeh, Y., Mehdizadeh Gazooei, Y. & Molaei, S. (2013). Use of bioinformatics tools to study exon 2 of GDF9 gene in Tali and Beetal goats. Modern Genetics Journal (MGJ), 8(334), 283-288.
  11. Hadizadeh, M., Niazi, A., Mohammad Abadi, M., Esmailizadeh, A. & Mehdizadeh Gazooei, Y. (2014). Bioinformatics analysis of the BMP15 exon 2 in Tali and Beetal goats. Modern Genetics, 9(1), 117-120
  12. Jeffrey, J. S., Nolan, L. K., Tonooka, K. H. & et al. (2012). Virulence factors of Escherichia coli from cellulitis or colisepticemia lesions in chickens. Avian Diseases, 46, 48-52
  13. Johnson, J. R., Kuskowski, M. A., Gajewski, A., Soto, S., Horcajada, J. P., Jimenez, M. T., de Anta, H. & Vila, J. (2013). Extended virulence genotypes and phylogenetic background of Escherichia coli isolates from patients with cystitis, pyelonephritis, or prostatitis. The Journal of Infectious Diseases, 191, 46-50.
  14. Kawano, M., Yaguchi, K. & Osawa, R. (2016). Genotypic analyses of Escherichia coli isolated from chickens with colibacillosis and apparently healthy chickens in Japan. Microbiol Immunol, 51, 961-966.
  15. McPeak, S. J. W., Smyth, J. A. &Ball, H. J. (2005). Characterisation of avian pathogenic Escherichia coli (APEC) associated with colisepticaemia compared to fecal isolates from healthy bird. Veterinary Microbiology, 110,245-253.
  16. Moazeni, S., Mohammd Adabadi, M. R., Sadeghi, M., Moradi Shahrbabak, H., Koshkoieh, A. & Bordbar, F. (2016a). Association between UCP Gene Polymorphisms and Growth, Breeding Value of Growth and Reproductive Traits in Mazandaran Indigenous Chicken. Open Journal of Animal Sciences, 6(1), 1-8.
  17. Moazeni, S. M., Mohammadabadi, M. R., Sadeghi, M., Moradi Shahrbabak, H. & Esmailizadeh, A. K. (2016). Association of the melanocortin-3 (MC3R) receptor gene with growth and reproductive traits in Mazandaran indigenous chicken. Journal of Livestock Science and Technologies, 4(2), 51-56.
  18. Mohammadabadi, M. R., Nikbakhti, M., Mirzaee, H. R., Shandi, A., Saghi, D. A., Romanov, M. N. & Moiseyeva, I. G. (2010). Genetic variability in three native Iranian chicken populations of the Khorasan province based on microsatellite markers. Russian Journal of Genetics, 46(4), 505-509.
  19. Mohammadifar, A. & Mohammadabadi, M. R. (2017). The Effect of Uncoupling Protein Polymorphisms on Growth, Breeding Value of Growth and Reproductive Traits in the Fars Indigenous Chicken. Iranian Journal of Applied Animal Science, 7(4), 679-685.
  20. Murakamin, M., Kwaga, J. K., White, D. G. & et al. (2006). Escherichia coli cellulitis in broiler chickens: clonal relationships among strains and analysis of virulence-associated factors of isolates from diseased birds. Infect Immun, 64, 3118-3126.
  21. Murray, C., Ratcliff, R., Cameron, P. & Dixon, S. (2014). The resistance of antimicrobial agents in salmonella from veterinary sources in Australia from 1995-2011. Australian Veterinary, 63(9), 286-291.
  22. Murray, P. R., Kobayashi, G. S., Pfaller, M. A. & Rosenthal Ken, S. (2010). Medical Microbiolog    (2nd Ed.). International Edition, 227-232.
  23. Ragion, R. M., Sayers, A. R. & Woodward, M. J. (2009). The role of fimbriae and flagella in the in clonization, invasion and persistence of Escherichia coli O78: K80 in the day-old chick model. Epidemiology and Infection,124, 351-363.
  24. Ramazanzadeh, R., Farhadifar, F. & Mansouri, M. (2010). Etiology and antibiotic resistance pattern of community-acquired extended-spectrum beta-lactamas-producing gram negative isolates in Sanandaj. Research Journal of Medical Sciences, 4, 243-7.
  25. Sader, H. S. (2016). Antimicrobial activity of tetracycline tested against nosocomial bacterial pathogens from patients hospitalized in the intensive care unit.Diagnostic Microbiology and Infectious Disease, 52(3), 203-8.
  26. Shahdadnejad, N., Mohammadabadi, M. R. & Shamsadini, M. (2016). Typing of Clostridium Perfringens Isolated from Broiler Chickens Using Multiplex PCR. Genetics in the 3rd millennium, 14(4).
  27. Stordeur, P., Bree, A., Mainil, J. & Moulin-Schouleur, M. (2007). Pathogenicity of pap-negative avian Escherichia coli isolated from septicaemic lesions. Microbes Infect, 6, 637-645.
  28. Suwanich Kul, A., Paningrahy, B. & Wagner, M. (2009).Antigen relateness and partial amino acid sequence of pili of Escherichia coli serotype O1, O2 and O78 pathogenic to poultry. Avian Diseases, 31, 809-813.
  29. Van Bost, S., Jacquemin, E., Oswald, E. & Mainil, J. (2016). Multiplex PCRs for indentification of necrotoxigenic Escherichia coli. Clinical Microbiology, 41(9), 4480-4482.
  30. Zandi, E., Mohammadabadi, M. R., Ezzatkhah, M. & Esmailizadeh, A. K. (2014). Typing of Toxigenic Isolates of Clostridium Perfringens by Multiplex PCR in Ostrich. Iranian Journal of Applied Animal Science, 4, 509-514.
  31. West, B. & Zhou, B. X. (1989). Did Chicken go North? New Evidence for Domestication, World's Poultry Science Journal, 45, 205-218.