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Characterization of methicillin-resistant coagulase-negative staphylococci in milk from cows with mastitis in Brazil

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Abstract

Staphylococci are one of the most prevalent microorganisms in bovine mastitis. Staphylococcus spp. are widespread in the environment, and can infect animals and humans as opportunistic pathogens. The objective of this study was to determine the frequency of methicillin-resistance (MR) among coagulase-negative staphylococci (CoNS) previously obtained from milk of mastitic cows in Brazil and to characterize the antimicrobial resistance phenotype/genotype and the SCCmec type of MRCoNS isolates. Identification of MRCoNS was based on both biochemical and molecular methods. Susceptibility testing for eleven antimicrobials was performed by disk-diffusion agar. Antimicrobial resistance genes and SCCmec were investigated by specific PCRs. Twenty-six MRCoNS were detected (20 % of total CoNS), obtained from 24 animals, and were identified as follows: S. epidermidis (7 isolates), S. chromogenes (7), S. warneri (6), S. hyicus (5) and S. simulans (1). All MRCoNS isolates carried mecA while the mecC gene was not detected in any CoNS. The SCCmec IVa was demonstrated in nine MRCoNS, while the remaining 17 isolates harbored non-typeable SCCmec cassettes. In addition to oxacillin and cefoxitin resistance, MRCoNS showed resistance to tetracycline (n = 7), streptomycin (n = 6), tobramycin (n = 6), and gentamicin (n = 4), and harbored the genes tet(K) (n = 7), str (n = 3), ant(4′) (n = 6) and aac(6′)-aph(2″) (n = 4), respectively. In addition, seven strains showed intermediate resistance to clindamycin and two to streptomycin, of which two harboured the lnu(B) and lsa(E) genes and two the aad(E) gene, respectively. One isolate presented intermediate erythromycin and clindamycin resistance and harbored an erm(C) gene with an uncommon 89-bp deletion rendering a premature stop codon. MRCoNS can be implicated in mastitis of cows and they constitute a reservoir of resistance genes that can be transferred to other pathogenic bacteria.

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Acknowledgments

Silva N. C. C. has a fellowship from Capes–Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, Process–9877-11-8. P. Gomez has a predoctoral fellowship of the Universidad de La Rioja of Spain. Part of this work was financially supported by Project SAF2012-35474 from the Ministerio de Economía y Competitividad of Spain and Fondo Europeo de Desarrollo Regional (FEDER).

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Authors and Affiliations

  1. Department of Microbiology and Immunology, UNESP, Botucatu, SP, Brazil

    Nathalia C. C. Silva, Joao P. Araújo-Júnior & Vera L. M. Rall

  2. Department of Agri-food Industry, Food and Nutrition–LAN USP, Piracicaba, SP, Brazil

    Nathalia C. C. Silva

  3. Department of Hygiene Veterinary and Public Health, UNESP, Botucatu, SP, Brazil

    Felipe F. Guimarães, Marcela de P. Manzi & Helio Langoni

  4. Department of Food and Agriculture, Universidad de La Rioja, Área de Bioquímica y Biología Molecular, Madre de Dios 51, 26006, Logroño, Spain

    Elena Gómez-Sanz, Paula Gómez & Carmen Torres

  5. Environmental Genomics and Systems Biology Research Group, Institute of Natural Resource Sciences Zurich University of Applied Sciences (ZHAW), Wädenswil, Switzerland

    Elena Gómez-Sanz

Authors
  1. Nathalia C. C. Silva
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  2. Felipe F. Guimarães
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  3. Marcela de P. Manzi
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  4. Elena Gómez-Sanz
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  5. Paula Gómez
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  6. Joao P. Araújo-Júnior
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  7. Helio Langoni
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  8. Vera L. M. Rall
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  9. Carmen Torres
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Correspondence to Carmen Torres.

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Silva, N.C.C., Guimarães, F.F., de P. Manzi, M. et al. Characterization of methicillin-resistant coagulase-negative staphylococci in milk from cows with mastitis in Brazil. Antonie van Leeuwenhoek 106, 227–233 (2014). https://doi.org/10.1007/s10482-014-0185-5

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  • DOI: https://doi.org/10.1007/s10482-014-0185-5

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