Detection of three virulence genes in E.coli isolates from commercial broilers with colibacillosis and their antibiotic resistance profiles in Tabriz area, Iran

Document Type: Original Articles

Authors

Department of Avian Diseases, Research & Diagnosis, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization, Tehran, Iran

Abstract

Colibacillosis caused by E.coli is one of the most common illnesses in commercial chickens and induce high financial losses in the poultry industry due to morbidity, mortality and antibiotic expenses because of overuses. E.coli can mostly cause disease as a secondary or opportunistic pathogen, while certain strains with specific virulence factors even as primary pathogen can cause colibacillosis. The aim of this study was to detect the three virulence genes including irp2, papC, and tsh in E.coli bacteria isolated from diseased broilers in Tabriz area using by multiplex polymerase chain reaction (PCR) method. The other purpose of this study was to do the in vitro drug sensitivity of the isolates using by disk diffusion method. 38 (53.5%), 25 (35.2%) and 35 (49.3%) isolates out of total 71 isolates, showed, irp2, papC and tsh genes respectively. In 16 isolates only irp2 gene, 8 isolates only papC gene, and in 13 isolates only tsh gene were detected and 12 isolates were without all these genes. In 39 isolates 2 or 3 gene were identified simultaneously. In this study most of the isolates were resistant against enrofloxacin, doxycycline, sultrim, erythromycin and danofloxacin and most of them were sensitive only to colistin and the percentage of sensitive isolates against fosfomycin, neomycin and florfenicol were 45.9%, 45.2% and 48.3% respectively. High resistance rate to most antibiotics and prevalence of some important virulence genes in APEC (avian pathogenic Escherichia coli ) isolates might be a serious hazard for both poultry industry and public health.

Keywords

Main Subjects


Article Title [French]

Détection de 3 gènes virulents et des profils de résistances aux antibiotiques chez les isolats d’Escherichia coli issus de poulets de chair affectés par la colibacillose à Tabriz, Iran

Abstract [French]

La colibacillose causée par l’Escherichia coli est l’une des maladies les plus répandues chez les poulets de chair commerciaux et engendre des pertes financières considérables dans l’industrie avicole. Ces pertes sont principalement générées par les taux élevés de morbidité, de mortalité et de consommation d’antibiotiques liés à cette infection. Les maladies causées par l’E. colisont souvent induites par desagents pathogènes secondaires ou opportunistes, alors que certaines souches virulentes de cette bactérie peuvent agir comme pathogènes primaires. L’objectif de cette étude était de déterminer la présence des trois gènes de virulence et les profils de résistance aux antibiotiques des isolats d’E. coliprélevés à partir de poulets de chair affectés par la colibacillose à Tabriz (Iran). A cet effet, une méthode de réaction en chaîne de la polymérase (PCR) multiplex et des antibiogrammes standards par diffusion ont été respectivement utilisés. Selon nos résultats, parmi les 71 isolats bactériens analysés, 38 (53,5%), 25 (35,2%) et 35 (49,3%) cas comprenaient respectivement les gènes irp2 (iron-repressibleprotein), papC (pyelonephritis-associatedpili) et tsh (temperature-sensitive hemagglutinin). De plus, 16 isolats contenaient uniquement le gène irp2, 8 isolats présentaient seulement le gène papC, 13 isolats incluaient exclusivement le gène tsh, alors que 12 isolats ne portaient aucun des trois gènes. Enfin, dans 39 isolats, 2 à 3 gènes étaient simultanément présents. La plupart des isolats étaient résistants aux antibiotiques enrofloxacine, doxycycline, sultrim, erythromycine et danofloxacine et montraient une sensibilité à la colistine. Les fréquences respectives de susceptibilité des isolats à la fosfomycine, la néomycine et au florfénicol étaient de 45,9%, 45,2% et 48,3%. Par conséquent, le taux important de résistance aux antibiotiques observés dans cette étude et la prévalence de certains gènes de virulence chez les isolats de l’E. colipathogénique aviaire, peuvent constituer un risque sérieux pour l’industrie avicole et la santé publique.

Keywords [French]

  • Escherichia coli
  • Gènes de virulence
  • Poulet de chair
  • Colibacillose
  • Résistance antibiotique
  • Iran

Arabi, S., Jafarpour, M., Mirinargesi, M., Behjati Asl, S., Naghshbandi, R., Shabanpour, M., 2013. Molecular characterization of avian pathogenic Escherichia coli in broilers bred in Northern Iran. Glob Vet 10 382-386.

Barnes, L.M., Bentley, C.M., Dickson, A.J., 2003. Stability of protein production from recombinant mammalian cells. Biotechnol Bioeng 81, 631-639.

Bauer, A.W., Kirby, W.M., Sherris, J.C., Turck, M., 1966. Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol 45, 493-496.

Cherifi, A., Contrepois, M., Picard, B., Goullet, P., Orskov, I., Orskov, F., 1994. Clonal relationships among Escherichia coli serogroup O78 isolates from human and animal infections. J Clin Microbiol 32, 1197-1202.

Dozois, C.M., Dho-Moulin, M., Bree, A., Fairbrother, J.M., Desautels, C., Curtiss, R., 3rd, 2000. Relationship between the Tsh autotransporter and pathogenicity of avian Escherichia coli and localization and analysis of the Tsh genetic region. Infect Immun 68, 4145-4154.

Ewers, C., Janssen, T., Kiessling, S., Philipp, H.C., Wieler, L.H., 2005. Rapid detection of virulence-associated genes in avian pathogenic Escherichia coli by multiplex polymerase chain reaction. Avian Dis 49, 269-273.

Ghaniei, A., Peighambari, S.M., 2011. Antimicrobial susceptibility of one thousand bacterial isolates to five antibacterial agents commonly used in the Iranian poultry industry. Iran J Vet Med 6, 1-5.

Gophna, U., Oelschlaeger, T.A., Hacker, J., Ron, E.Z., 2001. Yersinia HPI in septicemic Escherichia coli strains isolated from diverse hosts. FEMS Microbiol Lett 196, 57-60.

Janben, T., Schwarz, C., Preikschat, P., Voss, M., Philipp, H.C., Wieler, L.H., 2001. Virulence-associated genes in avian pathogenic Escherichia coli (APEC) isolated from internal organs of poultry having died from colibacillosis. Int J Med Microbiol 291, 371-378.

Karch, H., Schubert, S., Zhang, D., Zhang, W., Schmidt, H., Olschlager, T., et al., 1999. A genomic island, termed high-pathogenicity island, is present in certain non-O157 Shiga toxin-producing Escherichia coli clonal lineages. Infect Immun 67, 5994-6001.

Khoshkhoo, P.H., Peighambari, S.M., 2005. Drug resistance patterns and plasmid profiles of Escherichia coli isolated from cases of avian collibacilosis. Iran J Vet Med 60, 97-105.

Kwon, S.-G., Cha, S.-Y., Choi, E.-J., Kim, B., Song, H.-J., Jang, H.-K., 2008. Epidemiological Prevalence of Avian Pathogenic Escherichia coli Differentiated by Multiplex PCR from Commercial Chickens and Hatchery in Korea. J Bacteriol Virol 38, 179-188.

Madadi, M.S., Ghaniei, A., Zare, P., Isakakroudi, N., 2014. Antimicrobial Susceptibility Pattern of Escherichia coli Isolates to Antibacterial Agents in Urmia, Iran. Int J Basic Sci Res 3  695-697.

Moon, B.M., Won, G.Y., Choi, Y.Y., Jin, J.K., Oh, I.J., Park, J.H., 2006. Isolation and characteristics of avian pathogenic Escherichia coli from birds associated with colibacillosis. Chulalongkorn Uni Fac Vet Sci, 26-29.

Nakazato, G., Campos, T.A.d., Stehling, E.G., Brocchi, M., Silveira, W.D.d., 2009. Virulence factors of avian pathogenic Escherichia coli (APEC). Pesquisa Veterinária Brasileira 29, 479-486.

Nateghi, F., Jafarpour, M., Nazemi, A., 2010. A Survey for detection of eight correlated genes of avian pathogenic Escherichia coli in Human Uropathogenic Escherichia coli. J Microb World 3, 169-176.

Nolan, L.K., Barnes, H.J., Vaillancourt, J.P., Aziz, T.A., Logue, C.M., 2013. Colibacillosis, In: Diseases of Poultry, Wiley- Blackwell Press, Ames, IA.

Pourbakhsh, S.A., Dho-Moulin, M., Bree, A., Desautels, C., Martineau-Doize, B., Fairbrother, J.M., 1997. Localization of the in vivo expression of P and F1 fimbriae in chickens experimentally inoculated with pathogenic Escherichia coli. Microb Pathog 22, 331-341.

Rahimi, M., 2013. Antibioresistance profile of avian pathogenic Escherichia coli isolates recovered from broiler chicken farms with colibacillosis in Kermanshah province, Iran. Glob Vet 10 447-452.

Smith, J.L., Drum, D.J., Dai, Y., Kim, J.M., Sanchez, S., Maurer, J.J., et al., 2007. Impact of antimicrobial usage on antimicrobial resistance in commensal Escherichia coli strains colonizing broiler chickens. Appl Environ Microbiol 73, 1404-1414.

Swayne, D.E., 1998. A Laboratory Manual for the Isolation and Identification of Avian Pathogens, American Association of Avian Pathologists, American Association of Avian Pathologists.

White, D.G., Piddock, L.J., Maurer, J.J., Zhao, S., Ricci, V., Thayer, S.G., 2000. Characterization of fluoroquinolone resistance among veterinary isolates of avian Escherichia coli. Antimicrob Agents Chemother 44, 2897-2899.

Won, G.-Y., Moon, B.-M., Oh, I.-G., Matsuda, K., Chaudhari, A.A., Hur, J., et al., 2009. Profiles of Virulence-associated Genes of Avian Pathogenic Escherichia coli Isolates from Chickens with Colibacillosis. The J of Poult Sci 46, 260-266.

Yang, H., Chen, S., White, D.G., Zhao, S., McDermott, P., Walker, R., et al., 2004. Characterization of multiple-antimicrobial-resistant Escherichia coli isolates from diseased chickens and swine in China. J Clin Microbiol 42, 3483-3489.

Zahraei Salehi, T., Farashi Bonab, S., 2006. Antibiotics Susceptibility Pattern of Escherichia coli Strains Isolated from Chickens with Colisepticemia in Tabriz Province, Iran. Int J Poult Sci 5 677-684.