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Genetic Determinants of Antibiotic Resistance in Staphylococcus aureus Isolates from Milk of Mastitic Crossbred Cattle

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Abstract

Staphylococcus aureus is one of the major causes of mastitis in dairy animals and its resistance against multiple antimicrobials always remains crucial concern. Present investigation was carried out to detect the distribution of antibiotic-resistant genes of S. aureus isolates. Isolates (128) of S. aureus from mastitic milk were collected, tested for antibiotics with disc-diffusion method, and resistant genes mecA, linA, msrA msrB, vatA, vatB, vatC ermA, ermC tetK, tetM and aacA-D were detected by PCR. The phenotypic antibiotics resistance percent in S. aureus isolates was classified as tetracycline (36.7), gentamycin (30.5), streptomycin (26.6), kanamycin (25.8) and penicillin G (22.7). All the isolates were susceptible to vancomycin. Among isolates, 10.2% were observed as methicillin-resistant. The distribution of antibiotic-resistant genes was linA (51.6) followed by msrB (46.1), tetK + M (34.4), msrA and aacA-D (26.6%). Different antibiotic-resistant genes combinations (mecA/linA-2; mecA/aacA-D/tetK/linA/msrB-3; mecA/linA/msrA/msrB-3; aacA-D/linA/msrA/msrB-4; aacA-D/linA/msrB-7; linA/msrA/msrB-10; tetK/linA/msrA/msrB-11; aacA/tetK/linA/msrB-12 isolates) were observed. All the isolates lacked amplification of vatA, vatB, ermA and ermC genes. Molecular typing resulted genetic variation in protein A (6–12 repeats) and coagulase genes (A–E patterns) were observed. Coagulase A and D genotypes were more prevalent in antibiotic-resistant isolates, while E, B and C in susceptible ones. The significant observation was the prevalence of methicillin-resistant S. aureus, which were resistant to multiple antibiotics. Findings revealed the status of resistant isolates in herd that might be helpful in treatment, controlling of resistant strains and culling of cows for mastitis reduction.

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References

  1. Aires-de-Sousa M, Parente CESR, Vieira-da-Motta O et al (2007) Characterization of Staphylococcus aureus isolates from buffalo, bovine, ovine, and caprine milk samples collected in Rio de Janeiro State, Brazil. Appl Environ Microbiol 73:3845–3849

    Article  CAS  PubMed  Google Scholar 

  2. Ardic N, Ozyurt M, Sarryyupoglu B et al (2005) Investigation of erythromycin and tetracycline resistance genes in methicillin-resistant staphylococci. Int J Antimicrob Agents 26:213–218

    Article  CAS  PubMed  Google Scholar 

  3. Boerlin P, Kuhnert P, Hüssy D, Schaellibaum M (2003) Methods for identification of Staphylococcus aureus isolates in cases of bovine mastitis. J Clin Microbiol 41:767–771

    Article  CAS  PubMed  Google Scholar 

  4. Chambers HF (1997) Methicillin resistance in staphylococci: molecular and biochemical basis and clinical implications. Clin Microbiol Rev 10:781–791

    CAS  PubMed  Google Scholar 

  5. CLSI (2008) Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals; Approved Standard-Third Edition (M31–A3). CLSI, Wayne

    Google Scholar 

  6. De Oliveira AP, Watts JL, Salmon SA et al (2000) Antimicrobial susceptibility of Staphylococcus aureus isolated from bovine mastitis in Europe and the United States. J Dairy Sci 83:855–862

    Article  PubMed  Google Scholar 

  7. Erskine RJ, Walker RD, Bolin CA et al (2002) Trends in antibacterial susceptibility of mastitis pathogens during a seven-year period. J Dairy Sci 85:1111–1118

    Article  CAS  PubMed  Google Scholar 

  8. Galdhar CN, Kumthekar SM, Samad A et al (2005) Current trends in diagnosis of mastitis in veterinary practice in India. Int J Cow Sci 1:77–79

    Google Scholar 

  9. Gentilini E, Denamiel G, Llorente P et al (2000) Antimicrobial susceptibility of Staphylococcus aureus isolated from bovine mastitis in Argentina. J Dairy Sci 83:1224–1227

    CAS  PubMed  Google Scholar 

  10. Gonano M, Hein I, Zangerl P et al (2009) Phenotypic and molecular characterization of Staphylococcus aureus strains of veterinary, dairy and human origin. Epidemiol Infect 137:688–699

    Article  CAS  PubMed  Google Scholar 

  11. Hendriksen RS, Mevius DJ, Schroeter A et al (2008) Prevalence of antimicrobial resistance among bacterial pathogens isolated from cattle in different European countries: 2002–2004. Acta Vet Scand 50:1–10

    Article  CAS  Google Scholar 

  12. Jayaraman R (2009) Antibiotic resistance: an overview of mechanisms and a paradigm shift. Curr Sci 96:1475–1484

    CAS  Google Scholar 

  13. Kwon NH, Park KT, Moon JS et al (2005) Staphylococcal cassette chromosome mec (SCCmec) characterization and molecular analysis for methicillin-resistant Staphylococcus aureus and novel SCCmec subtype Ivg isolated from bovine milk in Korea. J Antimicrob Chemother 56:624–632

    Article  CAS  PubMed  Google Scholar 

  14. Lee JH (2003) Methicillin (oxacillin)-resistant Staphylococcus aureus strains isolated from major food animals and their potential transmission to humans. Appl Environ Microbiol 69:6489–6494

    Article  CAS  PubMed  Google Scholar 

  15. Lina G, Quaglia A, Reverdy M et al (1999) Distribution of genes encoding resistance to macrolides, lincosamides, and streptogramins among Staphylococci. Antimicrob Agents Chemother 43:1062–1066

    CAS  PubMed  Google Scholar 

  16. Løvseth A, Loncarevic S, Berdal KG (2004) Modified multiplex PCR method for detection of pyrogenic exotoxin genes in staphylococcal isolates. J Clin Microbiol 42:3869–3872

    Article  PubMed  CAS  Google Scholar 

  17. Lüthje P, von Köckritz-Blickwede M, Schwarz S (2007) Identification and characterization of nine novel types of small staphylococcal plasmids carrying the lincosamide nucleotidyltransferase gene lnu(A). J Antimicrob Chem 59:600–606

    Article  CAS  Google Scholar 

  18. Makovec JA, Ruegg PL (2003) Antimicrobial resistance of bacteria isolated from dairy cow milk samples submitted for bacterial culture: 8, 905 samples (1994–2001). J Am Vet Med Assoc 222:1582–1589

    Article  CAS  PubMed  Google Scholar 

  19. Montesinos I, Salido E, Delgado T, Cuervo M, Sierra A (2002) Epidemiological genotyping of methicillin resistant Staphylococcus aureus by pulsed field gel electrophoresis at a university hospital and comparison with antibiotyping and protein A and coagulase gene polymorphisms. J Clin Microbiol 40:2119–2125

    Article  CAS  PubMed  Google Scholar 

  20. Moon JS, Lee AR, Kang HM et al (2007) Phenotypic and genetic antibiogram of methicillin-resistant staphylococci isolated from bovine mastitis in Korea. J Dairy Sci 90:1176–1185

    Article  CAS  PubMed  Google Scholar 

  21. Ochoa-Zarzosa A, Loeza-Lara PD, Torres-Rodríguez F et al (2008) Antimicrobial susceptibility and invasive ability of Staphylococcus aureus isolates from mastitis from dairy backyard systems. Antonie Leeuwenhoek 94:199–206

    Article  CAS  PubMed  Google Scholar 

  22. Rabello RF, Souza CR, Duarte RS et al (2005) Characterization of Staphylococcus aureus isolates recovered from bovine mastitis in Rio de Janeiro, Brazil. J Dairy Sci 88:3211–3219

    Article  CAS  PubMed  Google Scholar 

  23. Reinosoa EB, El-Sayed A, Lämmler C, Bognia C, Zschöck M (2008) Genotyping of Staphylococcus aureus isolated from humans, bovine subclinical mastitis and food samples in Argentina. Microbiol Res 163:314–322

    Article  CAS  Google Scholar 

  24. Sabour PM, Gill JJ, Lepp D et al (2004) Molecular typing and distribution of Staphylococcus aureus isolates in Eastern Canadian dairy herds. J Clin Microbiol 42:3449–3455

    Article  CAS  PubMed  Google Scholar 

  25. Schwarz S, Roberts MC, Werckenthin C et al (1998) Tetracycline resistance in Staphylococcus spp. from domestic animals. Vet Microbiol 63:217–227

    Article  CAS  PubMed  Google Scholar 

  26. Strommenger B, Kettlitz C, Werner G et al (2003) Multiplex PCR Assay for simultaneous detection of nine clinically relevant antibiotic resistance genes in Staphylococcus aureus. J Clin Microbiol 41:4089–4094

    Article  CAS  PubMed  Google Scholar 

  27. Sung JML, Lindsay JA (2007) Staphylococcus aureus strains that are hypersusceptible to resistance gene transfer from enterococci. Antimicrob Agents Chemother 51:2189–2191

    Article  CAS  PubMed  Google Scholar 

  28. Türkyilmaz S, Kaya O (2006) Determination of some virulence factors in Staphylococcus Spp isolated from various clinical samples. Turk J Vet Anim Sci 30:127–132

    Google Scholar 

  29. Turutoglu H, Hasoksuz M, Ozturk D et al (2009) Methicillin and aminoglycoside resistance in Staphylococcus aureus isolates from bovine mastitis and sequence analysis of their mecA genes. Vet Res Commun. doi: 10.1007/s11259-009-9313-5

  30. Tyagi A, Kapil A, Singh P (2008) Incidence of methicillin resistant Staphylococcus aureus (MRSA) in pus samples at a tertiary care hospital, AIIMS, New Delhi. J Indian Acad Clin Med 9:33–35

    Google Scholar 

  31. Van den Eede A, Martens A, Lipinska U et al (2009) High occurrence of methicillin-resistant Staphylococcus aureus ST398 in equine nasal samples. Vet Microbiol 133:138–144

    Article  PubMed  CAS  Google Scholar 

  32. Vintov J, Aarestrup FM, Zinn CE et al (2003) Association between phage types and antimicrobial resistance among bovine Staphylococcus aureus from 10 countries. Vet Microbiol 95:133–147

    Article  CAS  PubMed  Google Scholar 

  33. Wang Y, Wu CM, Lu LM et al (2008) Macrolide-lincosamide-resistant phenotypes and genotypes of Staphylococcus aureus isolated from bovine clinical mastitis. Vet Microbiol 130:118–125

    Article  CAS  PubMed  Google Scholar 

  34. Zmantar T, Chaieb K, Ben Abdallah F et al (2008) Multiplex PCR detection of the antibiotic resistance genes in Staphylococcus aureus strains isolated from auricular infections. Folia Microbiol 53:357–362

    Article  CAS  Google Scholar 

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Acknowledgement

This study was supported by a grant from the National Fellow Scheme of Indian Council of Agriculture Research, New Delhi, India. Authors are highly thankful to technical staff of Livestock Genome Analysis Laboratory for the help in this study.

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Authors and Affiliations

  1. Livestock Genome Analysis Laboratory, National Dairy Research Institute, Karnal, 132001, India

    Ravinder Kumar & B. R. Yadav

  2. Carbohydrate and Protein Biotechnology Laboratory, Department of Biotechnology, Punjabi University, Patiala, 147002, India

    Ravinder Kumar & R. S. Singh

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  1. Ravinder Kumar
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  2. B. R. Yadav
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Correspondence to B. R. Yadav.

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Kumar, R., Yadav, B.R. & Singh, R.S. Genetic Determinants of Antibiotic Resistance in Staphylococcus aureus Isolates from Milk of Mastitic Crossbred Cattle. Curr Microbiol 60, 379–386 (2010). https://doi.org/10.1007/s00284-009-9553-1

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