Journal of Biosciences

, Volume 36, Issue 1, pp 175–188 | Cite as

Antibiotic resistance and pathogenicity factors in Staphylococcus aureus isolated from mastitic Sahiwal cattle



Methicillin-resistant Staphylococcus aureus (MRSA) poses a serious problem in dairy animals suffering from mastitis. In the present study, the distribution of mastitic MRSA and antibiotic resistance was studied in 107 strains of S. aureus isolated from milk samples from 195 infected udders. The characterizations pathogenic factors (adhesin and toxin genes) and antibiotic susceptibility of isolates were carried out using gene amplification and disc diffusion assays, respectively. A high prevalence of MRSA was observed in the tested isolates (13.1%). The isolates were also highly resistant to antibiotics, i.e. 36.4% were resistant to streptomycin, 33.6% to oxytetracycline, 29.9% to gentamicin and 26.2% each to chloramphenicol, pristinomycin and ciprofloxacin. A significant variation in the expression of pathogenic factors (Ig, coa and clf) was observed in these isolates. The overall distribution of adhesin genes ebp, fib, bbp, fnbB, cap5, cap8, map and cna in the isolates was found to be 69.1, 67.2, 6.5, 20.5, 60.7, 26.1, 81.3 and 8.4%, respectively. The presence of fib, fnbB, bbp and map genes was considerably greater in MRSA than in methicillin-susceptible S. aureus (MSSA) isolates. The proportions of toxin genes, namely, hlb, seb, sec, sed, seg and sei, in the isolates were found to be 94.3, 0.9, 8.4, 0.9, 10.2 and 49.5%, respectively. The proportions of agr genes I, II, III and IV were found to be 39.2, 27.1, 21.5 and 12.1%, respectively. A few isolates showed similar antibiotic-resistance patterns, which could be due to identical strains or the dissemination of the same strains among animals. These findings can be utilized in mastitis treatment programmes and antimicrobials strategies in organized herds.


Antibiotic resistance cattle mastitis MRSA pathogenic genes 


  1. Aarestrup FM, Lartsen HD, Eriksen NH, Elsberg CS and Jensen NE 1999 Frequency of alpha- and beta-haemolysin in Staphylococcus aureus of bovine and human origin. A comparison between pheno- and genotype and variation in phenotypic expression. APMIS 107 425–430PubMedCrossRefGoogle Scholar
  2. Akineden Ö, Annemüller C, Hassan AA, Lämmler C, Wolter W and Zschöck M 2001 Toxin genes and other characteristics of Staphylococcus aureus isolates from milk of cows with mastitis. Clin. Diagn. Lab. Immunol. 8 959–964PubMedGoogle Scholar
  3. Annemüller C, Lammler CH and Zschöck M 1999 Genotyping Staphylococcus aureus isolated from bovine mastitis. Vet. Microbiol. 69 217–224PubMedCrossRefGoogle Scholar
  4. Boerlin P, Kuhnert P, Hüssy D and Schaellibaum M 2003 Methods for identification of Staphylococcus aureus isolates in cases of bovine mastitis. J. Clin. Microbiol. 41 767–771PubMedCrossRefGoogle Scholar
  5. Booth MC, Pence LM, Mahasreshti P, Callegan MC and Gilmore MS 2001 Clonal associations among Staphylococcus aureus isolates from various sites of infection. Infect. Immun. 69 345–352PubMedCrossRefGoogle Scholar
  6. Brakstad OG, Aasbakk K and Maeland JA 1992 Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene. J. Clin. Microbiol. 30 1654–1660PubMedGoogle Scholar
  7. Brody T, Yavatkar AS, Lin Y, Ross J, Kuzin A, Kundu M, Fann Y and Odenwald WF 2008 Horizontal gene transfers link a human MRSA pathogen to contagious bovine mastitis bacteria. PLoS ONE 3 e3074PubMedCrossRefGoogle Scholar
  8. Clinical and Laboratory Standards Institute 2008 Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals.Google Scholar
  9. De Oliveira AP, Watts JL, Salmon SA and Aarestrup FM 2000 Antimicrobial susceptibility of Staphylococcus aureus isolated from bovine mastitis in Europe and the United States. J. Dairy Sci. 83 855–862PubMedCrossRefGoogle Scholar
  10. El-Sayed A, Alber J, Lämmler C, Jäger S, Wolter W and Castañeda-Vázquez H 2006 Comparative study on genotypic properties of Staphylococcus aureus isolated from clinical and sub clinical mastitis in Mexico. Vet. Mex. 37 165–179Google Scholar
  11. Erskine RJ, Walker RD, Bolin CA, Bartlett PC and White DG 2002 Trends in antibacterial susceptibility of mastitis pathogens during a seven-year period. J. Dairy Sci. 85 1111–1118PubMedCrossRefGoogle Scholar
  12. Fitzgerald JR, Sturdevant DE, Mackie SM, Gill SR and Musser JM 2001 Evolutionary genomics of Staphylococcus aureus: insights into the origin of methicillin-resistant strains and the toxic shock syndrome epidemic. Proc. Natl. Acad. Sci. USA 98 8821–8826PubMedCrossRefGoogle Scholar
  13. Fournier B 2008 Global regulators of Staphylococcus aureus virulence genes; in Staphylococcus molecular genetics (ed.) JA Lindsay (Norfolk: Caister Academic Press) pp 131–183Google Scholar
  14. Gentilini E, Denamiel G, Llorente P, Godaly S, Rebuelto M and DeGregorio O 2000 Antimicrobial susceptibility of Staphylococcus aureus isolated from bovine mastitis in Argentina. J. Dairy Sci. 83 1224–1227PubMedCrossRefGoogle Scholar
  15. Gilot P and van Leeuwen W 2004 Comparative analysis of agr locus diversification and overall genetic variability among bovine and human Staphylococcus aureus isolates. J. Clin. Microbiol. 42 1265–1269PubMedCrossRefGoogle Scholar
  16. Kumar R, Yadav BR and Singh RS 2010 Genetic Determinants of antibiotic resistance in Staphylococcus aureus isolates from milk of mastitic crossbred cattle. Curr. Microbiol. 60 379–386PubMedCrossRefGoogle Scholar
  17. Lammers A, Nuijten JM and Smith E 1999 The fibronectin binding proteins of Staphylococcus aureus are required for adhesion to and invasion of bovine mammary gland cells. FEMS Microbiol. Lett. 180 103–109PubMedCrossRefGoogle Scholar
  18. Lee JH 2003 Methicillin (oxacillin)-resistant Staphylococcus aureus strains isolated from major food animals and their potential transmission to humans. Appl. Environ. Microbiol. 69 6489–6494PubMedCrossRefGoogle Scholar
  19. Lina G, Boutite F, Tristan A, Bes M, Etienne J and Vandenesch F 2003 Bacterial competition for human nasal cavity colonization: role of staphylococcal agr alleles. Appl. Environ. Microbiol. 69 18–23PubMedCrossRefGoogle Scholar
  20. Løvseth A, Loncarevic S and Berdal K G 2004 Modified multiplex PCR method for detection of pyrogenic exotoxin genes in staphylococcal isolates. J. Clin. Microbiol. 42 3869–3872PubMedCrossRefGoogle Scholar
  21. Makovec JA and Ruegg PL 2003 Antimicrobial resistance of bacteria isolated from dairy cow milk samples submitted for bacterial culture: 8905 samples (1994–2001). J. Am. Vet. Med. Assoc. 222 1582–1589PubMedCrossRefGoogle Scholar
  22. Mehrotra M, Wang G and Johnson WM 2000 Multiplex PCR for detection of genes for Staphylococcus aureus enterotoxins exfoliative toxins toxic shock syndrome toxin 1 and methicillin resistance. J. Clin. Microbiol. 38 1032–1035PubMedGoogle Scholar
  23. Montesinos I, Salido E, Delgado T, Cuervo M and 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 2119–2125PubMedCrossRefGoogle Scholar
  24. Moon JS, Lee AR, Kang HM, Lee ES, Kim MN, Paik YH, Park YH, Joo YS and Koo HC 2007 Phenotypic and genetic antibiogram of methicillin-resistant staphylococci isolated from bovine mastitis in Korea. J. Dairy Sci. 90 1176–1185PubMedCrossRefGoogle Scholar
  25. Moore PCL and Lindsay JA 2001 Genetic variation among hospital isolates of methicillin-sensitive Staphylococcus aureus: evidence for horizontal transfer of virulence genes J. Clin. Microbiol. 39 2760–2767PubMedCrossRefGoogle Scholar
  26. Murakami K, Minamide W, Wada K, Nakamura E, Teraoka H and Watanabe S 1991 Identification of methicillin-resistant strains of staphylococci by polymerase chain reaction. J. Clin. Microbiol. 29 2240–2244PubMedGoogle Scholar
  27. Palma M, Haggar A and Flock JI 1999 Adherence of Staphylococcus aureus Is enhanced by an endogenous secreted protein with broad binding activity. J. Bacteriol. 181 2840–2845PubMedGoogle Scholar
  28. Peacock SJ, Moore CE, Justice A, Kantzanou M, Story L, Mackie K, O’Neill G and Day NPJ 2002 Virulent combinations of adhesins and toxin genes in natural populations of Staphylococcus aureus. Infect. Immun. 70 4987–4996PubMedCrossRefGoogle Scholar
  29. Phonimdaeng P, O’Reilly M, Nowlan P, Bramley AJ and Foster TJ 1990 The coagulase of Staphylococcus aureus 8325–4 sequence analysis and virulence of site-specific coagulase deficient mutants. Mol. Microbiol. 4 393–404PubMedCrossRefGoogle Scholar
  30. Reinoso EB, El-Sayed A, Lämmler C, Bognia C and Zschöck M 2008 Genotyping of Staphylococcus aureus isolated from humans bovine subclinical mastitis and food samples in Argentina. Microbiol. Res. 163 314–322PubMedCrossRefGoogle Scholar
  31. Sabour PM, Gill JJ, Lepp D, Pacan JC, Ahmed R, Dingwell R and Leslie K 2004 Molecular typing and distribution of Staphylococcus aureus isolates in Eastern Canadian dairy herds. J. Clin. Microbiol. 42 3449–3455PubMedCrossRefGoogle Scholar
  32. Salasia SIO, Khusnan Z, Lämmler C and Zschöck M 2004 Comparative studies on pheno- and genotypic properties of S. aureus isolated from bovine subclinical mastitis in central java in Indonesia and Hesse in Germany. J. Vet. Sci. 5 103–109PubMedGoogle Scholar
  33. Seaman P, Day M, Denver Russell A and Ochs D 2004 Susceptibility of capsular Staphylococcus aureus strains to some antibiotics, triclosan and cationic biocides. J. Antimicrob. Chemother. 4 696–698CrossRefGoogle Scholar
  34. Seki K, Sakurada J, Seong HK, Murai M, Tachi H, Ishii H and Masuda S 1998 Occurrence of coagulase serotype among Staphylococcus aureus strains isolated from healthy individuals-special reference to correlation with size of protein-A gene. Microbiol. Immunol. 42 407–409PubMedGoogle Scholar
  35. Stephan R, Annemüller C, Hassan AA and Lämmler C 2001 Characterization of enterotoxigenic Staphylococcus aureus strains isolated from bovine mastitis in north-east Switzerland. Vet. Microbiol. 78 373–382PubMedCrossRefGoogle Scholar
  36. Tristan A, Ying L, Bes M, Etienne J, Vandenesch F and Lina G 2003 Use of multiplex PCR to identify Staphylococcus aureus adhesins involved in human hematogenous infections. Clin. Microbiol. 41 4465–4467CrossRefGoogle Scholar
  37. Turutoglu H, Hasoksuz M, Ozturk D, Yildirim M and Sagnak S 2009 Methicillin and aminoglycoside resistance in Staphylococcus aureus isolates from bovine mastitis and sequence analysis of their mecA genes. Vet. Res. Commun. 33 945–956PubMedCrossRefGoogle Scholar
  38. Van den Eede A, Martens A, Lipinska U, Struelens M, Deplano A, Denis O, Haesebrouck F, Gasthuys F and Hermans K 2009 High occurrence of methicillin-resistant Staphylococcus aureus ST398 in equine nasal samples. Vet. Microbiol. 133 138–144PubMedCrossRefGoogle Scholar
  39. Vintov J, Aarestrup FM, Zinn CE and Olsen JE 2003 Association between phage types and antimicrobial resistance among bovine Staphylococcus aureus from 10 countries. Vet. Microbiol. 95 133–147PubMedCrossRefGoogle Scholar
  40. Wang Y, Wu CM, Lu LM, Ren GWN, Cao XY and Shen JZ 2008 Macrolide-lincosamide-resistant phenotypes and genotypes of Staphylococcus aureus isolated from bovine clinical mastitis. Vet. Microbiol. 130 118–125PubMedCrossRefGoogle Scholar
  41. Zecconi A, Binda E, Borromeo V and Piccinini R 2005 Relationship between some Staphylococcus aureus pathogenic factors and growth rates and somatic cell counts. J. Dairy Res. 72 203–208PubMedCrossRefGoogle Scholar
  42. Zschöck M, Risse K and Sommerhauser J 2004 Occurrence and clonal relatedness of sec/tst-gene positive Staphylococcus aureus isolates of quarter milk samples of cows suffering from mastitis. Lett. Appl. Microbiol. 38 493–498CrossRefGoogle Scholar

Copyright information

© Indian Academy of Sciences 2011

Authors and Affiliations

  1. 1.Livestock Genome Analysis Laboratory, National Dairy Research InstituteKarnalIndia
  2. 2.Carbohydrate and Protein Biotechnology Laboratory, Department of BiotechnologyPunjabi UniversityPatialaIndia

Personalised recommendations