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Identification of virulence factors by multiplex PCR in Escherichia coli isolated from calves in Minas Gerais, Brazil

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Abstract

In this study, multiplex PCR was employed to investigate the virulence factors of Escherichia coli strains isolated from 60-day-old calves. Faecal samples were collected from 54 calves at 12 dairy farms in the state of Minas Gerais, Brazil. A total of 156 isolates were obtained after culture and microbiological isolation and were tested by multiplex PCR for the presence of genes encoding toxins (Stx1, Stx2 and STa) and adherence factors (intimin, F41 and F5). Seventy of 156 isolates were positive for at least one virulence factor: ten (14.3 %) from diarrhoeic animals and 60 (85.7 %) from healthy calves. The virulence markers identified were: Stx1 (82.8 %), eae (24.3 %), F41 (11.4 %), F5 (10 %), STa (4.28 %) and Stx2 (4 %). In diarrhoeic animals, Stx1 (70 %) and F41 (30 %) were identified, while Stx1 (83.3 %), eae (28.3 %), F41 (8.3 %), F5 (11.6 %), STa (5 %) and Stx2 (1.6 %) were detected in isolates from healthy calves. Mixed infections with pathotypes Shiga toxin-producing E. coli (STEC)/enteropathogenic E. coli, STEC/enterohaemorrhagic E. coli and STEC/other (eae/F5, Stx1/STa) were detected in five healthy calves. Pathogenic E. coli were identified in 59.26 % of all calves and on 75 % of the dairy farms studied, not only in diarrhoeic (five of six) but also in healthy calves (27 of 48), which demonstrates the importance of this agent in the aetiology of diarrhoea in calves in the state of Minas Gerais.

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Acknowledgments

This work was supported by the Conselho Nacional de Desenvolvimento Técnico e Científico (CNPq) and Fundação de Amparo à Pesquisa do Estado de Minas Gerais. We thank the support of the Pró-Reitoria de Pesquisa da Universidade Federal de Minas Gerais (PRPq 04/2011). M.B.H. and A.P.L. have scholarships from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). F.M.C. is a fellowship recipient of the Coordenação de Aperfeiçoamento do Pessoal de Nível Superior (CAPES), Brazil. G.I.A. is CAPES ProDoc fellowship recipient.

Ethical standards

This study is in agreement with the Ethical Principles in Animal Experimentation (CETEA/UFMG, protocol number 145/2006).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Departamento de Medicina Veterinária Preventiva, Escola de Veterinária, Universidade Federal de Minas Gerais, UFMG, Av. Antônio Carlos, 6627, CEP30123-970, Belo Horizonte, Minas Gerais, Brazil

    Giovanna I. Andrade, Fernanda M. Coura, Ethiene L. S. Santos, Grazielle C. F. Galinari, Andrey P. Lage & Marcos B. Heinemann

  2. Departamento de Clínica e Cirurgia Veterinária, Escola de Veterinária, Universidade Federal de Minas Gerais, UFMG, Av. Antônio Carlos, 6627, CEP30123-970, Belo Horizonte, Minas Gerais, Brazil

    Marina G. Ferreira, Elias J. Facury Filho & Antônio U. de Carvalho

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  1. Giovanna I. Andrade
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  2. Fernanda M. Coura
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  3. Ethiene L. S. Santos
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  4. Marina G. Ferreira
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  9. Marcos B. Heinemann
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Correspondence to Marcos B. Heinemann.

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Andrade, G.I., Coura, F.M., Santos, E.L.S. et al. Identification of virulence factors by multiplex PCR in Escherichia coli isolated from calves in Minas Gerais, Brazil. Trop Anim Health Prod 44, 1783–1790 (2012). https://doi.org/10.1007/s11250-012-0139-8

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