02Jan 2019

INSIGHTS INTO VIRULENCE AND ANTIMICROBIAL RESISTANCE PLASMID ASSOCIATED GENES OF ESBL ESCHERICHIA COLI ASSOCIATED WITH ARTHRITIS IN CHICKENS IN EGYPT.

  • Bacteriology, Immunology and Mycology Department, Faculty of Veterinary Medicine, Benha University, Egypt.
  • Animal Health Research Institute, Tanta branch, Egypt.
  • Veterinary Hospital, Faculty of Veterinary Medicine, Zagazig University, 44511, Egypt.
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Increasing the incidence of extended spectrum beta-lactamase (ESBL) among Enterobacteriaceae especially avian pathogenic Escherichia coli (APEC) is of great concern to poultry industry. This study aimed to detect the prevalence and resistant profile of isolates showed ESBL activity among 100 E. coli isolates collected from hock joints in chickens (14-70 days old) with arthritis symptoms, from different chicken farms at EL-Gharbia Governorate in Egypt, in addition to, detect the incidence rates for selections of some virulence and antibiotic resistance plasmid associated genes using conventional PCR. A total 40 isolates were phenotypically identified as E. coli, with predominant (50%) O125 serotypes. All (100%) E. coli strains demonstrated a high level of multidrug resistance (MDR). Meanwhile, only 5 (12.5%) isolates were identified as ESBL producers based phenotypically on double disc synergy test and genotypically on screening for presence of β-lactamasesgenes. The ESBL isolates showed high resistance rates to β-lactam, aminoglycoside, tetracycline and quinolones antimicrobials, which could be explained by the presence of (100%) blaTEM, (100%) aadA, (80%) tetA and (20%) qnrS resistance genes, respectively. Regarding virulence genes, the result showed that only iss and ompA genes were detected in all isolates showed ESBL activity. The co-occurrence of these antibiotic resistance and virulence plasmid associated genes confirmed the acquisition of antimicrobial resistance genes by ESBL producing E. coli, which associated withincreased virulence. Thus, urgent intervention is needed to successfully control contamination with MDR isolates in Egypt especially in poultry and its products.


  1. Abd El-Mongy, M, G Abd-El-Moneam, AMoawad and A Mohammed, 2018. Serotyping and Virulence Genes Detection in Escherichia coli Isolated from Broiler Chickens. Journal of Biological Sciences. 18: 46-50.
  2. Ahmed, A M and T Shimamoto, 2011. Molecular characterization of antimicrobial resistance in Gram‐negative bacteria isolated from bovine mastitis in Egypt. Microbiology and immunology. 55: 318-327.
  3. Ahmed, O M, P Pangloli, C-A Hwang, S Zivanovic, T Wu, D D'souza and F A Draughon, 2015. The occurrence of Listeria monocytogenes in retail ready-to-eat meat and poultry products related to the levels of acetate and lactate in the products. Food Control. 52: 43-48.
  4. Al-Agamy, M H, 2013. Phenotypic and molecular characterization of extended-spectrum β-lactamases and AmpC β-lactamases in Klebsiellapneumoniae. Pakistan journal of pharmaceutical sciences. 26.
  5. Alekshun, M N and S B Levy, 2007. Molecular mechanisms of antibacterial multidrug resistance. Cell. 128: 1037-1050.
  6. Ammar, A, M El-Hamid, S Eid and A El Oksh, 2015. Insight into antimicrobial resistance and virulence genes of emergent multidrug resistant avian pathogenic Escherichia coli in Egypt: How closely related are they. Rev. Med. Vet. 166: 304-314.
  7. Bisi-Johnson, M A, C L Obi, S D Vasaikar, K A Baba and T Hattori, 2011. Molecular basis of virulence in clinical isolates of Escherichia coli and Salmonella species from a tertiary hospital in the Eastern Cape, South Africa. Gut pathogens. 3: 9.
  8. Blaak, H, R A Hamidjaja, A H van Hoek, L de Heer, A M de RodaHusman and F M Schets, 2014. Detection of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli on flies at poultry farms. Appl. Environ. Microbiol. 80: 239-246.
  9. Chen, R, W Schmidmayr, C Kr?mer, U Chen-Schmeisser and U Henning, 1980. Primary structure of major outer membrane protein II (ompA protein) of Escherichia coli K-12. Proceedings of the National Academy of Sciences. 77: 4592-4596.
  10. CLSI, 2011. Clinical and laboratory standards institute. Performance standards for antimicrobial susceptibility testing.
  11. CLSI, 2014. Clinical and Laboratory Standards Institute; Performance Standards for Antimicrobial Susceptibility Testing. 24th Informational Supplement M100-S24. CLSI, Wayne, PA, USA.
  12. Colom, K, J P?rez, R Alonso, AFern?ndez-Aranguiz, E Lari?o and R Cisterna, 2003. Simple and reliable multiplex PCR assay for detection of bla TEM, bla SHV and bla OXA?1 genes in Enterobacteriaceae. FEMS microbiology letters. 223: 147-151.
  13. Davin-Regli, A, J-M Bolla, C E James, J-P Lavigne, J Chevalier, E Garnotel and A Molitor, 2008. Membrane permeability and regulation of drug ?influx and efflux? in enterobacterial pathogens. Current drug targets. 9: 750-759.
  14. Dierikx, C, A van Essen-Zandbergen, K Veldman, H Smith and D Mevius, 2010. Increased detection of extended spectrum beta-lactamase producing Salmonella enterica and Escherichia coli isolates from poultry. Veterinary microbiology. 145: 273-278.
  15. Dutil, L, R Irwin, R Finley, L K Ng, B Avery, P Boerlin, A-M Bourgault, L Cole, D Daignault and ADesruisseau, 2010. Ceftiofur resistance in Salmonella entericaserovar Heidelberg from chicken meat and humans, Canada. Emerging infectious diseases. 16: 48.Edward, P and W Ewings, 1972. Identification of enterobacteriacae, Minneapolis. Burgess Publishing Co.
  16. Eid, S and AErfan, 2013. Characterization of E. coli associated with high mortality of poultry flocks. Assiut Vet. Med. J. 59: 51-61.
  17. El-Shazly, D, S Nasef, F Mahmoud and D Jonas, 2017. Expanded spectrum β?lactamase producing Escherichia coli isolated from chickens with colibacillosis in Egypt. Poultry science. 96: 2375-2384.
  18. El Tawab, AA, A M Ammar, S A Nasef and R M Reda, 2015. Prevalence of E. coli in diseased chickens with its antibiogram pattern. Benha Veterinary Medical Journal. 28: 224-230.
  19. Ewers, C, G Li, H Wilking, S Kieβling, K Alt, E-M Ant?o, C Laturnus, I Diehl, S Glodde and T Homeier, 2007. Avian pathogenic, uropathogenic, and newborn meningitis-causing Escherichia coli: how closely related are they? International Journal of Medical Microbiology. 297: 163-176.
  20. Finegold, S M and W T Martin, 1982. Diagnostic Microbiology 6th Ed. C.V. Mosby Co. St. Louis Toronto, Londan.
  21. Frankel, G, A D Phillips, I Rosenshine, G Dougan, J B Kaper and S Knutton, 1998. Enteropathogenic and enterohaemorrhagic Escherichia coli: more subversive elements. Molecular microbiology. 30: 911-921.
  22. Ghosal, A, R Bhowmick, R K Nandy, T Ramamurthy and N S Chatterjee, 2007. PCR-based identification of common colonization factor antigens of enterotoxigenic Escherichia coli. Journal of clinical microbiology. 45: 3068-3071.
  23. Gomis, S M, C Riddell, AA Potter and B J Allan, 2001. Phenotypic and genotypic characterization of virulence factors of Escherichia coli isolated from broiler chickens with simultaneous occurrence of cellulitis and other colibacillosis lesions. Canadian Journal of Veterinary Research. 65: 1.
  24. Hawkey, P M and A M Jones, 2009. The changing epidemiology of resistance. Journal of Antimicrobial Chemotherapy. 64: i3-i10.
  25. Jeong, Y-W, T-E Kim, J-H Kim and H-J Kwon, 2012. Pathotyping avian pathogenic Escherichia coli strains in Korea. Journal of veterinary science. 13: 145-152.
  26. Johnson, T J, Y Wannemuehler, C Doetkott, S J Johnson, S C Rosenberger and L K Nolan, 2008. Identification of minimal predictors of avian pathogenic Escherichia coli virulence for use as a rapid diagnostic tool. Journal of clinical microbiology. 46: 3987-3996.
  27. Khater, E S and H W Sherif, 2014. Rapid Detection of Extended Spectrum [Beta]-lactamase (ESBL) Producing Strain of Escherichia coli in Urinary Tract Infections Patients in Benha University Hospital, Egypt. British Microbiology Research Journal. 4: 443.
  28. Kwon, S-G, S-Y Cha, E-J Choi, B Kim, H-J Song and H-K Jang, 2008. Epidemiological prevalence of avian pathogenic Escherichia coli differentiated by multiplex PCR from commercial chickens and hatchery in Korea. Journal of Bacteriology and Virology. 38: 179-188.
  29. Laube, H, A Friese, C Von Salviati, B Guerra, AK?sbohrer, L Kreienbrock and U Roesler, 2013. Longitudinal monitoring of Esbl/Ampc-Producing Escherichia coli in German broiler chicken fattening farms. Applied and environmental microbiology. AEM. 00856-00813.
  30. Machado, E, R Cant?n, F Baquero, J-C Gal?n, ARoll?n, L Peixe and T M Coque, 2005. Integron content of extended-spectrum-β-lactamase-producing Escherichia coli strains over 12 years in a single hospital in Madrid, Spain. Antimicrobial agents and chemotherapy. 49: 1823-1829.
  31. Mulvey, M R, G Soule, D Boyd, W Demczuk, R Ahmed and M-p S T C C S Group, 2003. Characterization of the first extended-spectrum beta-lactamase-producing Salmonella isolate identified in Canada. Journal of clinical microbiology. 41: 460-462.
  32. Nataro, J P and J B Kaper, 1998. Diarrheagenicescherichia coli. Clinical microbiology reviews. 11: 142-201.
  33. Oh, J, M Kang, J Kim, B An, E Song, J Kim, E Shin, M Kim, J Kwon and Y Kwon, 2011. Characterization of Escherichia coli isolates from laying hens with colibacillosis on 2 commercial egg-producing farms in Korea. Poultry science. 90: 1948-1954.
  34. Orskov, I, F Orskov, B Jann and K Jann, 1977. Serology, chemistry, and genetics of O and K antigens of Escherichia coli. Bacteriological reviews. 41: 667.
  35. Quinn, P, M Carter, B Markey, W Donnoly and F Leonard, 2002. Veterinary microbiology and microbial disease. 166-1117 Osney Mead. Oxford first LTd, Registered at the United Kingdom.
  36. Rahman, M, M Samad, M Rahman and S Kabir, 2004. Bacterio-pathological studies on salmonellosis, colibacillosis and pasteurellosis in natural and experimental infections in chickens. Bangladesh Journal of Veterinary Medicine. 2: 1-8.
  37. Randall, L, S Cooles, M Osborn, L Piddock and M J Woodward, 2004. Antibiotic resistance genes, integrons and multiple antibiotic resistance in thirty-five serotypes of Salmonella enterica isolated from humans and animals in the UK. Journal of Antimicrobial Chemotherapy. 53: 208-216.
  38. Rasheed, B, 2011. Isolation and identification of bacteria causing arthritis in chickens. Iraqi Journal of Veterinary Sciences. 25: 93-95.
  39. Robicsek, A, J Strahilevitz, G A Jacoby, M Macielag, D Abbanat, C H Park, K Bush and D C Hooper, 2006. Fluoroquinolone-modifying enzyme: a new adaptation of a common aminoglycoside acetyltransferase. Nature medicine. 12: 83.
  40. Sahoo, T K, L Sahoo, L N Sarangi, S K Panda and H K Panda, 2012. Prevalence, isolation, characterisation and antibiogram study of pathogenic Escherichia coli from different poultry farms of Odisha. Journal of Advanced Veterinary Research. 2: 169-172.
  41. Saidi, B, P Mafirakureva and J Mbanga, 2012. Antimicrobial resistance of Escherichia coli isolated from chickens with colibacillosis in and around Harare, Zimbabwe. Avian diseases. 57: 152-154.
  42. Shaza, Z M, 2016. Advanced microbiological studies on the bacterial causes of respiratory affections of poultry. Master thesis Bacteriology Department, Faculty of veterinary medicine Beni-Suef University, Egypt.
  43. Smith, J L, P M Fratamico and N W Gunther, 2007. Extraintestinal pathogenic Escherichia coli. Foodborne pathogens and disease. 4: 134-163.
  44. Srinivasan, P, G A Balasubramaniam, T R G K Murthy and P Balachandran, 2013. Bacteriological and pathological studies of egg peritonitis in commercial layer chicken in Namakkal area. Asian Pacific journal of tropical biomedicine. 3: 988-994.
  45. Su, L-H, C Chu, A Cloeckaert and C-H Chiu, 2008. An epidemic of plasmids? Dissemination of extended-spectrum cephalosporinases among Salmonella and other Enterobacteriaceae. FEMS Immunology & Medical Microbiology. 52: 155-168.
  46. Tawfik, R G, S A Khalil, H F Ellakany and H A Torky, 2016. Mycoplasma Synoviae and other Associated Bacteria Causing Arthritis in Chicken. Alexandria Journal for Veterinary Sciences. 49.
  47. WHO, 2014. World Health Organization; Antimicrobial resistance: Global report on surveillance. Available at http://www.who.int/drugresistance/documents/surveillancereport/en/.
  48. Yaguchi, K, T Ogitani, R Osawa, M Kawano, N Kokumai, T Kaneshige, T Noro, K Masubuchi and Y Shimizu, 2007. Virulence factors of avian pathogenic Escherichia coli strains isolated from chickens with colisepticemia in Japan. Avian diseases. 51: 656-662.

[Ashraf A. Abd El-Tawab, Fatma I. El-Hofy, khalid I. Alekhnawy, Asmaa T.Talaie and Ahmed A. Hefny. (2019); INSIGHTS INTO VIRULENCE AND ANTIMICROBIAL RESISTANCE PLASMID ASSOCIATED GENES OF ESBL ESCHERICHIA COLI ASSOCIATED WITH ARTHRITIS IN CHICKENS IN EGYPT. Int. J. of Adv. Res. 7 (Jan). 174-182] (ISSN 2320-5407). www.journalijar.com


Ahmed A. Hefny
Ph. D. Microbiology, Veterinary Hospital, Faculty of Veterinary Medicine, Zagazig University, 44511, Egypt

DOI:


Article DOI: 10.21474/IJAR01/8310      
DOI URL: http://dx.doi.org/10.21474/IJAR01/8310