Pathotyping and evaluation of multidrug resistancepattern in E.coli strains isolated from chicken meat retail stores in Shiraz

Document Type : Full Research Paper

Authors

Department of Microbiology, Jahrom Branch, Islamic Azad University, Jahrom, Iran.

Abstract

Escherichia coli is one of the most important members of the Enterobacteriaceae family and is a factor in causing generalized bacillosis in birds, followed by gastrointestinal upset in humans. The aim of this study was to investigate the antibiotic resistance and pathotyping of Escherichia coli isolated from chicken meat stores in Shiraz. In this cross-sectional descriptive study, the rectum of 183 chickens offered in chicken meat shops was sampled. After phenotypic diagnosis, 16S rRNA specific primers were used for genotypic confirmation.  Antibiogram test was performed for Escherichia coli samples by using disk diffusion method based on CLSI standard tables.Then, with PCR test and using specific primers of eae, LT, aatA, ipaH, stx genes, different pathotypes were isolated from each other. Using phenotypic and genotypic tests, 100(55%)  Escherichia coli samples were isolated. The frequencies of isolated pathotypes were EPEC (34%), ETEC (26%), EIEC (23%), EHEC (10%) and EAEC (7%), respectively. Antibiogram testing of Escherichia coli isolates showed that the highest antibiotic resistance was related to tetracycline (83%) and the lowest resistance in all pathotypes was related to gentamicin (10%). Also, the study of multiple resistance patterns showed that ETEC pathotype had the highest multiple drug resistance with 96%. The results of this study confirmed the transmission of various Escherichia coli pathotypes from farm animals to humans. Therefore, correct and timely diagnosis and treatment can prevent their spread in the community.

Keywords


1-Abe, C., F.A. Salvador, I.N. Falsetti, M.A.M. Vieira, J.Blanco, J.E. Blanco, M.Blanco, A.M.O. Machado, W.P. Elias, R.T. Hernandes and T.A.T. Gomes.2008. UropathogenicEscherichia coli (UPEC) strainsmay carry virulenceproperties of diarrhoeagenicE. Coli. FEMS ImmunologyandMicrobiology 52: 397–406.
2-Akbari Nakhjavani, F., M.Emaneini, , H.Hosseini, H. Iman-Eini,  M. Aligholi, F. Jabalameli, M.T.Haghi-Ashtiani, M.Taherikalaniand A.Mirsalehian. 2012. Molecular analysis of typical and atypical Enteropathogenic Escherichia coli (EPEC) isolated from children with diarrhea. Journal of  Medical Microbiology 62: 191-195.[In Farsi]
3-Aranda, K.R.S., U.Fagundes-Netoand I.C.A.Scaletsky. 2004. Evaluation of Multiplex PCRs for diagnosis of infection with  diarrheagenicEscherichia coli and Shigella spp.Journal of Clinical Microbiology42: 5849–5853.
4-Asadi, S., N.Shamsand M.NayebAghaei. 2017. Isolation and Frequency of Escherichia coli O157:H7 from Fecal Samples ofLorestan Ostrich Farms using Culture and Multiplex PCR. Journal of Veterinary Microbiology 13: 97-105.[In Farsi]
5-Asadi, A., T. Zahraei-Salehiand M.Jamshidian.2018.Molecular study of antibiotic resistance genes in Escherichia coli isolates in broilers of Shahrbabak city by multiplex PCR.Iranian Animal Sciences 49: 203-211.[In Farsi]
6- Bonkoungou,  I. J. O., T. Lienemann, O. Martikainen, R. Dembelé, I. Sanou, A. S. Traoré, A. Siitonen, N. Barro and K. Haukka. 2012. Diarrhoeagenic Escherichia coli detected by 16-plex PCR in children with and without diarrhoea in Burkina Faso.Clinical Microbiology and Infectious18:901-6. 
7-Bonyadian, M., H.Moshtaghiand P.Behrozi.2017. Occurrence of verotoxigenic E.coli in cow feces and antimicrobial resistance of the isolates in cattle farms in Shahrekord area .Journal of Microorganism Biology 6: 84-75.[In Farsi]
8- Clarke, L., B.Cherie millarand J.E. Moore. 2003. Extraction of genomic DNA from Pseudomonas aeruginosa: a comparison of three methods.Biomedical Science 60:34-35.
9- De Jesus, A.A.B., A.A.R.Freitas, J.C.de Souza, N.Martins, L.A.B.Botelho, V.B.C.Girão, L.M.Teixeira, L.Woodland Rileyand B.M.Moreira. 2018. High-Level Multidrug-Resistant Escherichia coli Isolates from Wild Birds in a Large Urban Environment.Microbial Drug Resistance25:167-172. 
10-Dutta.S., S. Guin, S.Ghosh, G.P. Pazhani, K.Rajendran, M.K. Bhattacharya, Y.Takeda, G.B.Nair and T.Ramamurthy.2013. Trends in the Prevalence of Diarrheagenic Escherichiacoli among Hospitalized Diarrheal Patients in Kolkata,India. PLoS ONE 8: 1-5
11- Foxman, B. 2014. Urinary Tract Infection Syndromes Occurrence, Recurrence, Bacteriology, Risk Factors,
and Disease Burden.Infectious Disease Clinics of North America28:1–13.
12- Hasan,B.,L.Sandegren,  A.Melhus, M.Drobni,J.Hernandez,J.Waldenström, M.Alam and B.Olsen. 2012.Antimicrobial Drug–Resistant Escherichia coli in Wild Birds and Free-range Poultry.Emerging Infectious Diseases18:2055-2058.
13-Hemmerum, A.M and O.E, Heuer. 2009. Human Health Hazards from Antimicrobial-ResistantEscherichia coli of Animal Origin. Clinical Infectious Diseases48:916-921.
14-Imuta.N, J. Nishi, K. Tokuda, R. Fujiyama, K. Manago, M. Iwashita, J.Sarantuya, and Y. Kawano.2008. The Escherichia coli Efflux Pump TolC Promotes Aggregation ofEnteroaggregative E. coli 042. Infection and immunity76:1247–1256.
15-Kang, H.Y., Y.S.Jeong,J.Y. Oh,  S.H.Tae, C.H. Choi,  D.C.Moon, W.K. Lee, Y.C. Lee,  S.Y.Seol, D.T. Choand J.C. Lee. 2005.Characterization of antimicrobial resistance and class 1 integrons found in Escherichia coli isolates from humans and animals in Korea.Journal of antimicrobial Chemotherapy55:639-44. 
16-Kimpe, A., A. Decostere, A. Martel, F. Haesebrouck, and L. A. Devriese. 2014. Prevalence of antimicrobial resistance amongpigeon isolates of Streptococcus gallolyticus ,Escherichia coli and Salmonella enterica serotypeTyphimurium. Avian Pathology 31: 393-397.
17- Haghighi Khoshkhoo, P. andI. Ali-Nezhad. 2010.Antibacterial Resistance Patterns of Escherichia coli Isolated from Broilers Colibacillosis of Broilers chicks in Golestan Province.Journal of Veterinary Clinical Reasearch1:39-47.[In Farsi]
18-Livermore,D.M.2008. Defining an extended-spectrum b-lactamase. Clinical Microbiology and Infection
14 : 3–10.
19-Madic.J, C. Peytavin de Garam, N. Vingadassalon, E. Oswald, P. Fach, E. Jametand F. Auvray. 2010. Simplex and multiplex real-time PCR assays for thedetection of flagellar (H-antigen) fliC alleles and intimin(eae) variants associated with enterohaemorrhagicEscherichia coli (EHEC) serotypes O26:H11, O103:H2,O111:H8, O145:H28 and O157:H7.Journal of Applied Microbiology109:1696-1705
20- Monte, D.F.,A.Mem, M.R.Fernandes, L.Cerdeira, F. Esposito, J.A.Galvão, B.D.G.M.Franco, N.Lincopanand M.Landgraf. 2017.Chicken Meat as Reservoir of Colistin-Resistant Escherichia coli Carrying mcr-1 Genes in South America.Antimicrobial Agents and Chemotherapy61: 1-4.
21-Mehrani Far, Z., M.Salehiand K.Amini. 2016.
 Detection of bla TEM, bla OXA and bla SHV genes in Escherichia coli isolated from colibacillosis in poultry by multiplex-PCR.Veterinary Clinical Pathology10: 81-90.[In Farsi]
22-Mitchel, N., J.Johnson, B. Johnston, R.Curtissand M.Mellata. 2015.Zoonotic potential of Escherichia coli isolates from retail chicken meat products and eggs.Applied and environmental microbiology81:1177-87.
23-Mohamed, L.,Z. Ge,  L.Yuehua,  G.Yubin,  K.Rachid,  O.Mustapha,  W.JunweiandO. Karine. 2018. Virulence traits of avian pathogenic (APEC) and fecal (AFEC) E. coli isolated from broiler chickens in Algeria.Tropical animal health and production50:547-553. 
24-Patel, J.B., M.P.Weinstein and G.M. Eliopoulos.2018.Zone Diameter and MIC Breakpoints for Enterobacteriaceae, pp.30-37, Performance Standards for Antimicrobial Susceptibility Testing in: M100 28th ed., CLSI, Pennsylvania, USA
25-Soleimanifard, N. and K.Amini. 2016. Molecular identification of EAEC and EPEC pathotypes of Escherichia coli isolated from cow's milk with mastitis by multiplex PCR and determination of antibiotic resistance by disk diffusion method and E.test method.
Journal of comparative pathobiology
 Iran 13:1849-1854.