Methicillin-resistant Staphylococcus aureus (MRSA) poses a serious problem in dairy animals suffering from mastitis. The study was carried out to evaluate the incidence of Methicillin resistant S. aureus from clinical mastitis milk samples and their antibiotic resistance profile and characterised with respect to the molecular features that contributed to the resistance in these pathogens. Isolation and identification of Methicillin resistant S. aureus were performed from acute clinical mastitis samples. The isolates were tested using agar disc diffusion method for their antimicrobial susceptibility and modified resazurin assay micro dilution technique for MIC to 8 different antimicrobial drugs. A total of 235 clinical mastitis milk samples from dairy cows were cultured for incidence of S. aureus. Methicillin resistant S. aureus was isolated from a total of 12 (44.25%) of the 116 S. aureus samples. Based on the antimicrobial sensitivity and MIC results, MRSA isolates were found sensitive to gentamicin, enrofloxcain, amoxicillin+sulbactam, ceftriaxone and resistant to amoxicillin, oxytetracycline, penicillin G and oxacillin. Most of MRSA isolates were found to be multi-drug resistant. MRSA alert kit test and mecA and blaZ target gene PCR were found to be useful in the confirmation of MRSA.
Antibiotic susceptibility, Antimicrobial drug resistance, Bovine mastitis, MIC, mecA, MRSA, PCR blaZ, PVL gene
Chambers, H,F. (1997). Methicillin resistance in staphylococci: Molecular and biochemical basis and clinical implications. Clinical Microbiology Review, 10: 781 - 791.
Clinical and Laboratory Standard Institute (CLSI). (2008). Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from Animals. pp. 65-72. In: Approved standard M31-A3 (3rd edn). Clinical and Laboratory Standards Institute, Wayne, PA.
Febler, A., Scott, C., Kadlec, K., Ehricht, R., Monecke, S. and Schwarz, S. (2010). Characterization of methicillin -resistant Staphylococcus aureus ST398 from cases of bovine mastitis. Journal of Antimicrobial Chemotherapy, 65(4): 619-25.
Juhász-Kaszanyitzky, E., Janosi, S., Somogyi, P., Dan, A., Bloois, L. G. and Wagenaar, J. A. (2007). MRSA transmission between cows and humans. Emerging Infectious Disease,13: 630–632.
Kenar, B., Kuyucuoglu, Y. and Seker, E. (2012). Antibiotic susceptibility of coagulase - negative staphylococci isolated from bovine subclinical mastitis in Turkey. Pakistan Veterinary Journal, 32: 390-393.
Kumar, R., Yadav, B. R. and Singh, R. S. (2010). Genetic determinants of antibiotic resistance in Staphylococcus aureus isolates from milk of mastitic crossbred cattle. Current Microbiology, 60: 379–386.
Kumar, R., Yadav, B. R. and Singh, R. S. (2011). Antibiotic resistance and pathogenicity factors in Staphylococcus aureus isolated from mastitic Sahiwal cattle. Journal of Biological Sciences, 36: 175-188.
Lee, J. H. (2003). Methicillin (oxacillin)-resistant Staphylococcus aureus strains isolated from major food animals and their potential transmission to humans. Applied Environmental Microbiology, 69: 6489–6494.
Lee, J. H., Jeong, J. M., Park, Y. H., Choi, S. S., Kim, Y. H., Chae, J. S., Moon, J. S., Kim, S. and Eo, S. K. (2004). Evaluation of the methicillin-resistant Staphylococcus aureus (MRSA)-screen latex agglutination test for detection of MRSA of animal origin. Journal of Clinical Microbiology, 42:2780–2782.
Lina, G., Piemont, Y., Godail-Gamot, F., Bes, M., Peter, M. O., Gauduchon, V. (1999). Involvement of pantonvalentine leukocidin-producing Staphylococcus aureus in primary skin infections and pneumonia. Clinical Infectious Disease, 29:1128-32.
Loeffler, A. and Lloyd, D. H. (2010). Companion animals: a reservoir for methicillin - resistant Staphylococcus aureus in the community?. Epidemiology of Infection, 138(5): 595-605.
Martineau, F., Picard, F. J., Lansac, N., Ménard, C., Roy, P. H., Ouellette, M. and Bergeron, M. G. (2000). Correlation between the resistance genotype determined by multiplex PCR assays and the antibiotic susceptibility patterns of Staphylococcus aureus and Staphylococcus epidermidis. Antimicrobial Agents Chemotherapy, 44: 231-238.
Olsen, J. E., Christensen, H. and Aarestrup, F. M. (2006). Diversity and evolution of blaZ from Staphylococcus aureus and coagulase negative Staphylococci. Journal of Antimicrobial Chemotherapy, 57: 450-460.
Piddock, L. J. V. (1996). Does the use of antimicrobial agents in veterinary medicine and animal husbandry select antibiotic resistant bacteria that infect man and compromise antimicrobial chemotherapy? Journal of Antimicrobial Chemotherapy, 38: 1-3.
Rankin, S., Roberts, S., O'shea, K., Maloney, D., Lorenzo, M. and Benson, C. E. (2005). Panton valentine leukocidin (PVL) toxin positive MRSA strains isolated from companion animals. Veterinary Microbiology, 108: 145-8.
Radostits, O. M., Blood, D. C. and Gay, G. C. (2008). Veterinary Medicine. A Textbook of the Diseases of cattle, sheep, pigs, goats and horses. 10th Ed., Bailliare Tindall, London.
Sarker, S. D., Nahar, L. and Kumarasamy, Y. (2007). Microtitre plate-based antibacterial assay incorporating resazurin as an indicator of cell growth, and its application in the in vitro antibacterial screening of phytochemicals. Methods, 42: 321-324.
Selepak, S. T. and Witebsky, F. G. (1985). Inoculum size and lot to lot variation as significant variables in the tube coagulase test for Staphylococcus aureus. Journal of Clinical Microbiology, 22:835-837.
Turutoglu, H., Ercelik, S. and Ozturk, D. (2006). Antibiotic resistance of Staphylococcus aureus and Coagulasenegative Staphylococci isolated from bovine mastitis Bulletin of Veterinary Institute in Pulawy, 50: 41-45.
Udo, E. E., Al-Obaid, I. A., Jacob, L. E. and Chugh, T. D. (1996). Molecular characterization of epidemic ciprofloxacin- and methicillin resistant Staphylococcus aureus strains colonizing patients in an intensive care unit. Journal of Clinical Microbiology, 34: 3242–3244.
Vandenesch, F., Naimi, T., Enright, M. C., Lina, G., Nimmo, G. R. and Hefferman, H. (2003). Community- acquired methicillin-resistant Staphylococcus aureus carrying Panton-Valentine leukocidin genes: worldwide emergence. Emerging Infectious Disease, 9: 978–984.
Van Loo, I., Huijsdens, X., Tiemersma, E., De Neeling, A., Van de Sande-Bruinsma, N., Beaujean, D. (2007). Emergence of methicillin-resistant Staphylococcus aureus of animal origin in humans. Emerging Infectious Disease, 13:1834-9.
Witte, W., Strommenger, B., Stanek, C. and Cuny, C. (2007). Methicillin-resistant Staphylococcus aureus ST398 in humans and animals, central Europe. Emerging Infectious Disease,13: 255-258.
Yugueros, J., Temprano, A., Sanchez, M., Luengo, J. and Naharro, G. (2001). Identification of Staphylococcus spp. by PCR-restriction fragment length polymorphism of gap gene. Journal of Clinical Microbiology, 39:3693–3695.
This work is licensed under Attribution-NonCommercial 4.0 International (CC BY-NC 4.0) © Author (s)