Advertisement

Virulence-associated genes and antimicrobial resistance among avian pathogenic Escherichia coli from colibacillosis affected broilers in Pakistan

  • Mariya Azam
  • Mashkoor MohsinEmail author
  • Sajjad-ur-Rahman
  • Muhammad Kashif Saleemi
Regular Articles
  • 51 Downloads

Abstract

Avian pathogenic Escherichia coli (APEC) causes colibacillosis that leads to high morbidity and mortality among poultry birds. To date, there is a lack of knowledge about virulence-associated genes (VAGs) and multidrug resistance of APEC isolates from Pakistan. In this study, we determined the VAGs and antibiotic resistance profiles of APEC isolates recovered from colibacillosis affected broilers in Faisalabad region of Pakistan. A total of 84 diseased and dead birds from different local broilers farms were collected and examined for the gross lesions of colibacillosis by conducting postmortem examination. Of these, APEC isolates were recovered from 75 (89.2%) birds. Antibiotic susceptibility tests against 11 antimicrobial agents showed the highest resistance against ampicillin (98.6%) followed by tetracycline (97.3%) and ciprofloxacin (72%). The presence of 11 virulence-associated genes (VAGs) was detected by multiplex polymerase chain reaction (PCR). Of the 75 APEC, 32 (42.6%) harbored > 5 VAGs. Most commonly found genes were increased serum survival (iss; 84%), iron transport (iutA; 74.6%), and colicin V (ColV; 60%). Twenty-two isolates (29.3%) were found to possess a combination of VAGs; iss, tsh, iroN, and iutA, in addition to other VAGs. To the best of our knowledge, this is the first report on the detection of virulence-associated genes and multidrug resistance among APEC isolates in Pakistan. In the future, the strains with the predominant set of VAGs can be used for colibacillosis diagnosis and as a potential vaccine candidate.

Keywords

Poultry Colibacillosis Avian pathogenic E. coli Virulence genes Antimicrobial resistance 

Notes

Funding

This work was supported by the fund of the International Foundation for Science, Sweden (IFS B/5793-1).

Compliance with ethical standards

Ethical approval

Ethical approval was obtained from the Institute of Biosafety Committee (IBC), University of Agriculture, Faisalabad, Pakistan. The animal samples were processed according to animal research guidelines approved by research ethics committee of Department of Pathology, University of Agriculture, Faisalabad, Pakistan. All authors have seen and approved the content and have contributed considerably to the work.

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Akond, M., 2009. Antibiotic resistance of Escherichia Coli isolated from poultry and poultry environment of Bangladesh, American Journal of Environmental Sciences, 5, 47–52CrossRefGoogle Scholar
  2. Azam, M., Ehsan, I., Sajjad-ur-Rahman, Saleemi, M.K., Javed, M.R. and Mohsin, M., 2017. Detection of the colistin resistance gene mcr-1 in avian pathogenic Escherichia coli in Pakistan, Journal of Global Antimicrobial Resistance, 11, 152–153Google Scholar
  3. Barbieri, N.L., Matter, B., Regina, S., Pinheiro, S., Ibelli, M., Nolan, L.K., Logue, C.M. and Guimara, B., 2015. Molecular characterization and clonal relationships among Escherichia coli strains isolated from broiler chickens with colisepticemia, 12, 74–83Google Scholar
  4. CLSI, 2012. Performance standards for antimicrobial disk susceptibility tests ; approved atandard Clinical and Laboratory Standars Institute - NCCLS, 32.Google Scholar
  5. Díaz-Sánchez, S., Sánchez, S., Ewers, C. and Höfle, U., 2012. Occurrence of avian pathogenic Escherichia coli and antimicrobial-resistant E. coli in red-legged partridges ( Alectoris rufa ): sanitary concerns of farming, Avian Pathology, 41, 337–344CrossRefGoogle Scholar
  6. Dissanayake, D.R.A., Octavia, S. and Lan, R., 2014. Population structure and virulence content of avian pathogenic Escherichia coli isolated from outbreaks in Sri Lanka, Veterinary Microbiology, 168, 403–412CrossRefGoogle Scholar
  7. Dou, X., Gong, J., Han, X., Xu, M., Shen, H., Zhang, D. and Zhuang, L., 2015. Characterization of avian pathogenic Escherichia coli isolated in eastern China, Gene, 576, 244–248CrossRefGoogle Scholar
  8. Ewers, C., Janssen, T., Kiessling, S., Philipp, H.C. and Wieler, L.H., 2005. Rapid detection of virulence-associated genes in avian pathogenic Escherichia coli by multiplex polymerase chain reaction, Avian Diseases, 49, 269–273CrossRefGoogle Scholar
  9. Filho, H.C.K., Brito, K.C.T., Cavalli, L.S. and Brito, B.G., 2015. Avian Pathogenic Escherichia coli ( APEC ) - an update on the control. The battle against microbial pathogens: Basic science, technological advances and educational programs, 598–618Google Scholar
  10. Guabiraba, R. and Schouler, C., 2015. Avian colibacillosis : still many black holes, FEMS, Microbiology Letters, 1–8Google Scholar
  11. Eid, H., M. Algamma, A., A. Nasef, S., Elfeil, W.K. and H. Mansour, G., 2016. Genetic variation among avian pathogenic E. coli strains isolated from broiler chickens, Asian Journal of Animal and Veterinary Advances, 11, 350–356Google Scholar
  12. Jeong, Y.-W., Kim, T.-E., Kim, J.-H. and Kwon, H.-J., 2012. Pathotyping avian pathogenic Escherichia coli strains in Korea, Journal of Veterinary Science, 13, 145CrossRefGoogle Scholar
  13. Jiang, H.X., L, D.H., Chen, Z.L., Wang, X.M., Chen, J.R., Liu, Y.H., Liao, X.P., Liu, J.H. and Zeng, Z.L., 2011. High prevalence and widespread distribution of multi-resistant Escherichia coli isolates in pigs and poultry in China, Veterinary Journal, 187, 99–103CrossRefGoogle Scholar
  14. Johnson, T.J., Siek, K.E., Johnson, S.J. and Nolan, L.K., 2005. DNA Sequence and Comparative Genomics of pAPEC-O2-R , an Avian Pathogenic Escherichia coli Transmissible R Plasmid, Antimicrobial agents and chemotherapy, 49, 4681–4688CrossRefGoogle Scholar
  15. Johnson, T.J., Wannemuehler, Y., Doetkott, C., Johnson, S.J., Rosenberger, S.C. and Nolan, L.K., 2008. Identification of minimal predictors of avian pathogenic Escherichia coli virulence for use as a rapid diagnostic tool, Journal of Clinical Microbiology, 46, 3987–3996CrossRefGoogle Scholar
  16. Kemmett, K., Williams, N.J., Chaloner, G., Humphrey, S., Wigley, P. and Humphrey, T., 2014. The contribution of systemic Escherichia coli infection to the early mortalities of commercial broiler chickens, Avian Pathology, 43, 37–42CrossRefGoogle Scholar
  17. Kim, T.E., Jeong, Y.W., Cho, S.H., Kim, S.J. and Kwon, H.J., 2007. Chronological study of antibiotic resistances and their relevant genes in Korean avian pathogenic Escherichia coli isolates, Journal of Clinical Microbiology, 45, 3309–3315CrossRefGoogle Scholar
  18. Li, Y., Chen, L., Wu, X. and Huo, S., 2013. Molecular characterization of multidrug-resistant avian pathogenic Escherichia coli isolated from septicemic broilers, Poultry Science, 94, 601–611CrossRefGoogle Scholar
  19. Lutful Kabir, S.M., 2010. Avian colibacillosis and salmonellosis: A closer look at epidemiology, pathogenesis, diagnosis, control and public health concerns, International Journal of Environmental Research and Public Health, 7, 89–114CrossRefGoogle Scholar
  20. Lv, J., Mohsin, M., Lei, S., Srinivas, S., Talish, R. and Lin, J., 2018. Discovery of a mcr-1- bearing plasmid in commensal colistin-resistant Escherichia coli from healthy broilers in Faisalabad , Pakistan, Virulence, 9, 994–999CrossRefGoogle Scholar
  21. Mbanga, J. and Nyararai, Y.O., 2015. Virulence gene profiles of avian pathogenic Escherichia coli isolated from chickens with colibacillosis in Bulawayo, Zimbabwe, The Onderstepoort journal of veterinary research, 82, E1–8Google Scholar
  22. Mohamed, M.A., Shehata, M.A. and Rafeek, E., 2014. Virulence genes content and antimicrobial resistance in escherichia coli from broiler chickens, Veterinary Medicine International, 1–6Google Scholar
  23. Mohamed, L., Ge, Z., Yuehua, L., Yubin, G., Rachid, K., Mustapha, O., Junwei, W. and Karine, O., 2018. Virulence traits of avian pathogenic (APEC) and fecal (AFEC) E. coli isolated from broiler chickens in Algeria, Tropical Animal Health and Production, 50, 547–553CrossRefGoogle Scholar
  24. Mohsin, M., Raza, S., Roschanski, N., Guenther, S., Ali, A. and Schierack, P., 2017. Description of the first Escherichia coli resistance Gene mcr-1 from the Indian sub continent, Antimicrobial Agents and Chemotherapy, 61, 1–2CrossRefGoogle Scholar
  25. Nhung, T. N., Chansiripornchai, N, Carrique, J. J., 2017. Antimicrobial resistance in bacterial poultry pathogens: A review, Frontiers in Veterinary Sciences, 4, 126CrossRefGoogle Scholar
  26. Obeng, A.S., Rickard, H., Ndi, O., Sexton, M. and Barton, M., 2012. Antibiotic resistance, phylogenetic grouping and virulence potential of Escherichia coli isolated from the faeces of intensively farmed and free range poultry, Veterinary Microbiology, 154, 305–315CrossRefGoogle Scholar
  27. Olesen, B., 2017. Characterization of four Escherichia coli clonal groups, Acta Pathologica Microbiologica et Immunologica Scandinavica, 125, 1–28CrossRefGoogle Scholar
  28. Paixao, A. C., Ferreira A.C., M. Fontes, P. Themudo, T. Albuquerque, M.C. Soares, M. Fevereiro, L. Martins, M.I. Corr and S. De., 2016. Detection of virulence-associated genes in pathogenic and commensal avian Escherichia coli isolates, Poultry Science, 95,1646–1652.CrossRefGoogle Scholar
  29. Rocha, A.C.G.P., Rocha, S.L.S., Lima-Rosa, C.A. V, Souza, G.F., Moraes, H.L.S., Salle, F.O., Moraes, L.B. and Salle, C.T.P., 2008. Genes associated with pathogenicity of avian Escherichia coli (APEC) isolated from respiratory cases of poultry, Pesquisa Veterinaria Brasileira, 28, 183–186CrossRefGoogle Scholar
  30. Ronco, T., Stegger, M., Olsen, R.H., Sekse, C., Nordstoga, A.B., Pohjanvirta, T., Lilje, B., Lyhs, U., Andersen, P.S. and Pedersen, K., 2017. Spread of avian pathogenic Escherichia coli ST117 O78:H4 in Nordic broiler production, BMC Genomics, 18, 13Google Scholar
  31. Roseliza R., S. Khairani., Z. Zunita., M. Ramlan and E. Khoo, 2017. Phylogenetic grouping and virulence gene profiles of Escherichia Coli isolates from chicken, Malaysian Journal of Veterinary Research. 8, 65–74.Google Scholar
  32. Saleemi, M.K., M.F.U. Hussan, M.Z. Khan, A. Khan, R.Z. Abbas and A. Ahmad, 2014. Hematobiochemical effects of colistin administered intramuscularly in growing broiler birds, Pakistan Veterinary Journal, 34, 78–81.Google Scholar
  33. Schouler, C., Schaeffer, B., Brée, A., Mora, A., Dahbi, G., Biet, F., Oswald, E., Mainil, J., Blanco, J. and Moulin-Schouleur, M., 2012. Diagnostic strategy for identifying avian pathogenic Escherichia coli based on four patterns of virulence genes, Journal of Clinical Microbiology, 50, 1673–1678CrossRefGoogle Scholar
  34. Solà-Ginés, M., Cameron-Veas, K., Badiola, I., Dolz, R., Majó, N., Dahbi, G., Viso, S., Mora, A., Blanco, J., Piedra-Carrasco, N., González-López, J.J. and Migura-Garcia, L., 2015. Diversity of multi-drug resistant avian pathogenic Escherichia coli (APEC) Causing outbreaks of colibacillosis in broilers during 2012 in Spain, PLoS ONE, 10, 1–14CrossRefGoogle Scholar
  35. Subedi, M., Luitel, H., Devkota, B., Bhattarai, R.K., Phuyal, S. and Panthi, P., 2018. Antibiotic resistance pattern and virulence genes content in avian pathogenic Escherichia coli (APEC) from broiler chickens in Chitwan , Nepal, BMC Veterinary Research, 4–9Google Scholar
  36. Trampel, D.W., Wannemuehler, Y., Nolan, L.K., Trampel, D.W., Wannemuehler, A.C.Y. and B, L.K.N., 2007. Characterization of Escherichia coli isolates from peritonitis lesions in commercial laying hens, Avian Diseases, 51, 840–844CrossRefGoogle Scholar
  37. Vhm, L., Iq, S., Pdpm, D., Lev, R., Shs, R., Mal, L., Af, D.T. and Rmr, S., 2017. Genes of virulence and phylogenetic group in isolates of avian pathogenic Escherichia coli, IMedPub Journals, 9, 1–5Google Scholar
  38. Wook, K., Young, H., Kuk, H., Kim, W. and Seok, I., 2014. Virulence factors of uropathogenic Escherichia coli of urinary tract infections and asymptomatic bacteriuria in children, Journal of Microbiology, Immunology and Infection, 47, 455–461CrossRefGoogle Scholar
  39. Younis, G., Awad, A. and Mohamed, N., 2017. Phenotypic and genotypic characterization of antimicrobial susceptibility of avian pathogenic Escherichia coli isolated from broiler chickens, Veterinary World, 10, 1167–1172CrossRefGoogle Scholar
  40. Zhang H, Rehman MU, Li K, Luo H, Lan Y, Nabi F, Shahzad M, Huang S, Liu X, Mehmood K, Iqbal MK and Li J, 2017. Antimicrobial resistance of Escherichia coli isolated from tibetan piglets suffering from white score diarrhea, Pakistan Veterinary Journal, 37, 43–46Google Scholar
  41. Zhao, S., Maurer, J.J., Hubert, S., Villena, J.F. De, Mcdermott, P.F., Meng, J., Ayers, S., English, L. and White, D.G., 2005. Antimicrobial susceptibility and molecular characterization of avian pathogenic Escherichia coli isolates, Veterinary Microbiology, 107, 215–224CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Institute of MicrobiologyUniversity of AgricultureFaisalabadPakistan
  2. 2.Department of PathologyUniversity of AgricultureFaisalabadPakistan

Personalised recommendations