Molecular characterization of Enterotoxigenic Escherichia coli isolates harboring genetic elements mediating multiple-drug resistance

PDF (English)
DOI : https://doi.org/10.12681/jhvms.24164
M. KOHANSAL
Department of Medical Genetics, School of Medicine, Fasa University of Medical Sciences, Fasa, Iran. Department of Biology, Payame Noor University, Tehran, Iran.
A. NAJAFI
Department of Medical Genetics, Tehran University of Medical Sciences, Tehran, Iran.
R. VIDAL
Disciplinary Program of Microbiology, Institute of Biomedical Sciences, Faculty of Medicine, Universidad de Chile, Santiago, Chile
Résumé

Intensive antibiotics’ use in the management of the disease in neonate calves, a major economic concern in bovine industry, is one of the contributors to high levels of antibiotic resistance of pathogenic bacteria. The objective of this study was to investigate the antibiotic resistance patterns and the frequency of integrons classes among Enterotoxigenic Escherichia coli (ETEC) strains isolated from neonatal calf diarrhea (NCD) in South of Iran. 412 recto-anal mucosal swabs from diarrheic calves were analyzed by biochemical fingerprinting and for virulence genes by polymerase chain reaction (PCR). The isolates were examined for their susceptibility to a panel of 8 antibacterial agents using the Kirby-Bauer disc diffusion method. Finally, the frequency of integron classes was detected in multi-drug resistant (MDR) strains by polymerase chain reaction restriction fragment length polymorphism (PCR- RFLP). 194 out of 412 (47.09%) diarrheic fecal samples harbored E. coli and 35 (18%) of them were identified as ETEC. The drug susceptibility test showed that all isolates were resistant to erythromycin, penicillin and trimethoprim/sulfamethoxazole and more than 80% were resistant to ampicillin and chloramphenicol. All isolates were MDR. 17 out of 35 (48.57%) isolates were identified possessed class 1 integron. High prevalence of class 1 integron in ETEC isolates was mainly associated with multidrug resistance. Cefixime was the most effective antibiotic in vitro.

Article Details
  • Rubrique
  • Research Articles
Creative Commons License

Ce travail est disponible sous licence Creative Commons Attribution - Pas d’Utilisation Commerciale 4.0 International.

Authors who publish with this journal agree to the following terms:

· Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution Non-Commercial License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.

· Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g. post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.

· Authors are permitted and encouraged to post their work online (preferably in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work.

Téléchargements
Les données relatives au téléchargement ne sont pas encore disponibles.
Références
Abraham Sam, Darren J. Trott, David Jordan, David M. Gordon, Mitchell D. Groves, John M. Fairbrother, Matthew G. Smith, Ren Zhang, and Toni A Chapman (2014) Phylogenetic and molecular insights into the evolution of multidrug-resistant porcine enterotoxigenic Escherichia coli in Australia. Int J Antimicrob Agents, 44: 105-111.
Acha S J, Kühn I, Jonsson P, Mbazima, G, Katouli M, Möllby R (2004) Studies on calf diarrhoea in Mozambique: prevalence of bacterial pathogens. J Appl Microbiol, 45(1), 27.
Acres S D (1985) Enterotoxigenic Escherichia coli infections in newborn
calves: a review. J. Dairy Sci. 68(1), 229-256.
Ahmed AM, Younis E E, Osman SA , Ishida Y, El-khodery SA, Shimamoto
T (2009) Genetic analysis of antimicrobial resistance in Escherichia coli isolated from diarrheic neonatal calves. Vet Microbiol, 136, 397-402.doi: 10.1016/j. vetmic.2008.11.021.
Atlas RM (2010) Handbook of microbiological media. 4th ed. Boca Raton,
USA: CRC Press.
Azam H, Ghezeljeh S M, Mahmoud S (2015) Prevalence of class 1 and 2 integrons among the multidrug resistant uropathogenic strains of Escherichia coli. Asian Biomed, 9:49-54. doi: 10.5372/1905- 7415.0901.367.
Backx A, Smith HE, Essen van A, Veldman KT, Mevius DJ (2006) Characterization of integrons in E. coli isolates of Dutch veal calves and dairy cows. ISVEE, 11:724-730.
Bakhshi B, Najibi S, Sepehri-Seresht S (2014) Molecular characterization of enterohemorrhagic Escherichia coli isolates from cattle. J Vet Med Sci
:1195-1199. doi:10.1292/jvms.13- 0237.
Barigye R, Ablesh G, Lisa MP, Lynn PS, Darlene F (2012) Krogh, and Susan
Olet. Prevalence and antimicrobial susceptibility of virulent and avirulent
multidrug-resistant Escherichia coli isolated from diarrheic neonatal calves. Am J Vet Res, 12: 1944-1950.
Bass L, Liebert CA, Lee MD, Summers AO, White DG, Thayer SG, Maurer
J J (1999) Incidence and characterization of integrons, genetic elements
mediating multiple-drug resistance, in avianEscherichia coli. Antimicrob
Agents Ch. 43:2925-9.
Bewick V, Cheek L, Ball J (2003) Statistics review 8: Qualitative data–tests
of association. Critical care ) Crit Care( , 30;8:46. doi: 10.1186/cc2428 Boerlin P, Travis R, Gyles CL, Reid-Smith R, Lim NJ, Nicholson V, McEwen
SA, Friendship R, Archambault M (2005) Antimicrobial resistance and virulence genes of Escherichia coli isolates from swine in Ontario. AEM. 71: 6753-6761.
Cameron A, McAllister TA (2016) Antimicrobial usage and resistance in
beef production. J Anim Sci Biotechnol. 2016 Dec;7:68.
de Verdier K, Nyman A, Greko C, Bengtsson B (2012) Antimicrobial resistance and virulence factors in Escherichia coli from Swedish dairy calves. Acta Vet Scand. 54: 2.
Diaz-Mejia J J, Amabile-Cuevas CF, Rosas I, Souza V (2008) An analysis of the evolutionary relationships of integron integrases, with emphasis on the prevalence of class 1 integrons in Escherichia coli isolates from clinical and environmentalorigins. Microbiology, 154: 94-102. doi:10.1099/mic.0.2007/008649-0.
Diekema DJ, BootsMiller BJ, Vaughn TE, Woolson RF, Yankey JW, Ernst EJ, Flach SD, Ward MM, Franciscus CL, Pfaller MA, Doebbeling BN 2004 Antimicrobial resistance trends and outbreak frequency in United States hospitals. Clin Infect Dis: 38: 78-85.
Du X, Shen Z, Wu B, Xia S, Shen J (2005) Characterization of class 1 integrons-mediated antibiotic resistance among calf pathogenic Escherichia coli. FEMS Microbiol Lett. 2005;245:295-8. doi: 10.1016/j.femsle.03.021.
El-Seedy F R, Abed A H, Yanni H A, El-Rahman S A (2016) Prevalence of Salmonella and E. coli in neonatal diarrheic calves. BUJAS. 31;5:45-51. doi:10.1016/j.bjbas.2015.11.010.
El-Sokkary M M A, Abdelmegeed ES (2015) Characterisation of class 1 integron among Escherichia coli isolated from Mansoura University Hospitals in Egypt. Adv Microbiol.5: 269-277.doi: 10.4236/aim.2015.54025.
Friedrich T, Sven Rahmann S, Weigel W, Rabsch W, Fruth A, Ron E, Gunzer F, Dandekar T, Hacker J, Müller T, Dobrindt U (2010) High-throughput microarray technology in diagnostics of enterobacteria based on genomewide probe selection and regression analysis. BMC Genomic, 11: 591-612.
Guerra B, Junker E, Schroeter A, Malorny B, Lehmann S, Helmuth R (2003) Phenotypic and genotypic characterization of antimicrobial resistance in German Escherichia coli isolates from cattle, swine and poultry. J Antimicrob Chemother. 52:489-92.doi: 10.1093/jac/dkg362.
Gulsun S, Oguzoglu N, Inan A, Ceran N (2005) The virulence factors and
antibiotic sensitivities of Escherichia coli isolated from recurrent urinary
tract infections. Saudi Med J, ;26:1755-8.
Gular L, Gündüz K, Ok Ü ( 2008) Virulence factors and antimicrobial susceptibility of Escherichia coli isolated from calves in Turkey. Zoonoses
Public Health. 55:249-57.
Harada K, Asai T (2010) Role of antimicrobial selective pressure and secondary factors on antimicrobial resistance prevalence in Escherichia coli from food-producing animals in Japan. J Biomed Biotechnol. 180682.
doi: 10.1155/2010/180682.
Hariharan, H, Coles, M, Poole D, Page R (2004) Antibiotic resistance among enterotoxigenic Escherichia coli from piglets and calves from piglets and calves with diarrhea. Can Vet J. 45:605.
Ibrahim ME, Magzoub M A, Bilal NE, Hamid M E (2013) Distribution of Class I integrons and their effect on the prevalence of multi-drug resistant Escherichia coli clinical isolates from Sudan. Saudi Med J, 34:240-7.
Kang S, Lee D, Shin S, Ahn J, Yoo H (2005) Changes in patterns of antimicrobial susceptibility and class 1 integron carriage among Escherichia coli isolates. J Vet Sci. 6:201-5.
Karczmarczyk M, Walsh C, Slowey R, Leonard N, Fanning S (2011) Molecular characterization of multidrug-resistant Escherichia coli isolates from Irish cattle farms. Appl Environ Microbiol. 77(20):7121-7. doi:10.1128/AEM.00601-11.
Kargar M, Mohammadalipour Z, Doosti A, Lorzadeh S, Japoni-Nejad A (2014) High prevalence of class 1 to 3 integrons among multidrug-resistant diarrheagenic Escherichia coli in southwest of Iran. Osong Public Health Res, 5: 193-198.
Keyes K, Hudson C, Maurer J J, Thayer S, White DG, Lee M D (2000) Detection of Florfenicol Resistance Genes inEscherichia coli Isolated from Sick Chickens. Antimicrob agents cH, 44: 421-424.
Kohansal, M., & Asad, A. G. (2018). Molecular analysis of Shiga toxin-producing Escherichia coli O157: H7 and non-O157 strains isolated from
calves. Onderstepoort Journal of Veterinary Research, 85(1), 1-7.
Koneman EW, Allen SD, Janda W M, Schreckenberger P C, Mycology (1997) In Color Atlasand Textbook of Diagnostic Microbiology; 5 th edn., pp 983–1057 Philadelphia, PA: Lippincott Williams & Wilkins.
Lindblom G.B, Åhrén C, Changalucha J, Gabone R, Kaijser B, Nilsson LÅ, Sjögren E, Svennerholm AM, Temu M (1995) Campylobacter jejuni/coli and Enterotoxigenic Eschericia coli (ETEC) in Faeces from Children and Adults in Tanzania. Scand J Infect Dis, 27: pp.589-593.
Levin BR, Lipsitch M, Perrot V, Schrag S, Antia R, Simonsen L, Moore Walker N, Stewart FM, (1997) The population genetics of antibiotic resistance. Clin Infect Dis. 24 Suppl 1:S9-16.
Rusheeba M, Shah MI, Wani SA, Pandit F, Ahmad Dar P, Iqbal Mir M (2015) Prevalence, serodiversity and antibiogram of enterotoxigenic Escherichia coli (ETEC) in diarrhoeic calves and lambs of Kashmir valley (J&K), India. IJANS. 7: 477-481.
Mathai, E, Grape M, Kronval G (2004) Integrons and multidrug resistance
among Escherichia coli causing community‐acquired urinary tract infection in southern India. Apmis, 112: 159-164.
Magiorakos AP, Srinivasan A, Carey RB, Carmeli Y, Falagas ME, Giske CG,
Harbarth S, Hindler JF, Kahlmeter G, Olsson‐Liljequist B, Paterson DL (2012) Multidrug‐resistant, extensively drug‐resistant and pandrug‐resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. CMI. 18:268-81.
Maynard C, Fairbrother JM, Bekal S, Sanschagrin F, Levesque RC, Brousseau R, Masson L, Lariviere S, Harel J (2003) Antimicrobial resistance genes in enterotoxigenic Escherichia coli O149: K91 isolates obtained over a 23-year period from pigs. Antimicrob agents Ch. Oct 1;47(10):3214-21.
Moredo FA, Pineyro PE, Márquez GC, Sanz M, Colello R, Etcheverría A, Padola NL, Quiroga MA, Perfumo CJ, Galli L, Leotta GA (2015) Enterotoxigenic Escherichia coli subclinical infection in pigs: bacteriological and genotypic characterization and antimicrobial resistance profiles. Foodborne pathogens dis. Aug 1;12:704-11.
Nield B S, Holmes A J, Gillings MR, Recchia GD, Mabbutt BC, Nevalainen
K H, Stokes HW (2001) Recovery of new integron classes from environmental DNA. FEMS Microbiol Lett, 195: 59-65.
Orden J A, Cid D, Ruiz‐Santa‐Quiteria JA, Garcia S, Martinez S, De la Fuente R (2002). Verotoxin‐producing Escherichia coli (VTEC), enteropathogenic E. coli (EPEC) and necrotoxigenic E. coli (NTEC) isolated from healthy cattle in Spain. J Appl Microbiol, 93: 29-35.
Oteo J, Lázaro E, de Abajo FJ, Baquero F, Campos J, members of EARSS S.Antimicrobial-resistant invasive Escherichia coli, Spain. Emerg Infect Dis, 2005 Apr;11(4):546.
Prapasawat W, Intarapuk A, Chompook P, Nakajima C, Suzuki Y, Suthienkul O (2017) Antimcrobial resistance, intgron, virulence gene, and multilocus sequence typing of esherichia coli isolates from postweaning piglets with and without. Se Asian J Trop Med, 48: pp.1042-1055.
Prism G. 2005. 5. GraphPad Software. San Diego, CA, USA.
Pourtaghi-Shotorban H, Dahpahlevan V, Shaghayegh A, Badiee A (2011)
Molecular detection of diarrheagenic entrotoxigenic escherichia coli (ETEC) from calves fecal samples in Alborz province. 2011, J.et.Clin. Res 2: 33-38.
Rijavec M, Erjavec MS, Avguštin JA, Reissbrodt R, Fruth A, Križan-Hergouth V, Žgur-Bertok D (2006) High prevalence of multidrug resistance and random distribution of mobile genetic elements among uropathogenic Escherichia coli (UPEC) of the four major phylogenetic groups. Curr Microbiol. Aug 1;53:158-62.
Sahm D F, Thornsberry C, Mayfield DC, Jones M E, Karlowsky JA (2001) Multidrug-Resistant Urinary Tract Isolates ofEscherichia coli: Prevalence
and Patient Demographics in the United States in 2000. Antimicrob agents ch, 45: 1402-1406.
Shams Z, Tahamtan Y, Pourbakhsh A, Hosseiny MH, Kargar M, Hayati M
(2012) Detection of enterotoxigenic K99 (F5) and F41 from fecal sample
of calves by molecular and serological methods. Comp Clin Path. 2012 Aug 1;21:475-8.
Shahrani M, Dehkordi FS, Momtaz H (2014) Characterization of Escherichia coli virulence genes, pathotypes and antibiotic resistance properties in diarrheic calves in Iran. Biological Research. 2014 Dec;47:28.
Shaheen BW, Oyarzabal OA, Boothe DM (2010) The role of class 1 and 2
integrons in mediating antimicrobial resistance among canine and feline
clinical E. coli isolates from the US. Vet Microbiol. Aug 26;144:363-70.
Souto MS, Coura FM, Dorneles EM, Stynen AP, Alves TM, Santana JA, Pauletti RB, Guedes RM, Viott AM, Heinemann MB, Lage AP (2017) Antimicrobial susceptibility and phylotyping profile of pathogenic Escherichia coli and Salmonella enterica isolates from calves and pigs in
Minas Gerais, Brazil. Trop anim health pro . Jan 1;49:13-23.
Tahamtan Y, Fozi MA, Hosseini MH, Mostafaee T (2014) Molecular characterization of class 1 integrons and antibiotic resistance to E. coli fromcattle, sheep and goats. OJVR. 18:701-9.
Umpiérrez A, Acquistapace S, Fernández S, Oliver M, Acuña P, Reolón E,
Zunino P (2016) Prevalence of Escherichia coli adhesion-related genes in neonatal calf diarrhea in Uruguay. J Infect Dev Ctries, 10, 472-477.
van den Bogaard AE, Stobberingh EE (2000) Epidemiology of resistance
to antibiotics: links between animals and humans. Int J of antimicrolag, 14, 327-335.(2013):
Van Meervenne E, Boon N, Verstraete K, Devlieghere F, De Reu K, Herman L, Buvens G, Piérard D, Van Coillie E (2013) Integron characterization and typing of Shiga toxin-producing Escherichia coli isolates in Belgium. J medical microbiol, 62(5), pp.712-719.
Ventola CL (2015) The antibiotic resistance crisis: part 1: causes and threats. IJPT. 40(4):277-283.
Wani SA, Hussain I, Beg SA, Rather MA, Kabli ZA, Mir MA, Nishikawa Y(2013) Diarrhoeagenic Escherichia coli and salmonellae in calves and lambs in Kashmir: absence, prevalence and antibiogram. Rev sci tech Off int Epiz. Dec 1;32:1-7.
Wayne PA. (2012)a. Clinical and laboratory standards . Performance standards for antimicrobial disk susceptibility tests; approved standard. Approve Standard M02-A11. CLSI, 11th ed. PA, USA.
Wayne PA. (2012)b. Clinical and laboratory standards institute methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobicall. Approve Standard M07-A9, CLSI, 9th ed, PA, USA.
White PA, McIver CJ, Rawlinson WD (2001) Integrons and gene cassettes in theenterobacteriaceae. Antimicrobial agents and chemotherapy. AAC.
;45:2658-61.
Yadegari Z, Brujeni GN, Ghorbanpour R, Moosakhani F, Lotfollahzadeh S
(2019) Molecular characterization of enterotoxigenic Escherichia coli isolated from neonatal calves diarrhea. Vet Res Forum, 10: 73
Younis EE, Ahmed AM, El-Khodery SA, Osman SA, El-Naker YF (2009) Molecular screening and risk factors of enterotoxigenic Escherichia coli and Salmonella spp. in diarrheic neonatal calves in Egypt. Res Vet Sci 87 (3):373–379.
Yu HS, Lee JC, Kang HY, Jeong YS, Lee EY, Choi CH, Tae SH, Lee YC, Seol SY, Cho DT ( 2004) Prevalence of dfr genes associated with integrons and dissemination of dfrA17 among urinary isolates of Escherichia coli in Korea, J Antimicrob Chemoth, 53:pp.445-450.
Zhang W, Zhao M, Ruesch L, Omot A, Francis D (2007) Prevalence of virulence genes in Escherichia coli strains recently isolated from young pigs with diarrhea in the US. Vet Microbiol 123:145–152.