Aim: This study was conducted to determine the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in cattle and buffalo and to study their antibiotic resistance pattern.

Materials and Methods: A total of 136 samples (skin and nasal swab) from cattle and buffalo were collected. MRSA was identified by conventional bacterial culture techniques which were further confirmed by amplification of S. aureus-specific 16S rRNA by polymerase chain reaction (PCR). The isolates were further analyzed for the presence of mecA gene by PCR. The antimicrobial susceptibility profiling was performed by disc diffusion method.

Results: The prevalence of MRSA in the current study was 28.57% and 34.28% in cattle nasal and skin swab, respectively, with an overall prevalence of 31.43% MRSA among cattle. Buffalo nasal and skin sample showed MRSA prevalence of 54.55% and 39.4%, respectively, with 46.9% overall prevalence. PCR could detect mecA gene in 36.4% and 58% MRSA isolates from cattle and buffalo, respectively. Antimicrobial susceptibility test found MRSA resistant to penicillin and oxytetracycline (88% each), cefoxitin (75%), cotrimoxazole (62%), and amoxyclav (50%). 100% sensitivity was observed against ciprofloxacin, amikacin, chloramphenicol, and gentamicin. Three (16.7%) MRSA isolates from buffalo were found resistant to vancomycin.

Conclusion: Cattle and buffalo were identified as a potential carrier of MRSA in Bihar (India). The isolation of vancomycinresistant S. aureus (VRSA) in the current study indicates the emergence of VRSA in animal population which may be transmitted to the human beings working in close contact to the animals.

Keywords: antibiogram, bovine, mecA gene, methicillin resistant Staphylococcus aureus.

1. Vanderhaeghen, W., Herman, K., Haesebrouck, F. and Butaye, P. (2010) Methicillin resistant Staphylococcus aureus (MRSA) in food production animals. Epdemiol. Infect., 138: 606-625. [Crossref] [PubMed]

2. Wertheim, H.F.L., Lelles, D.C., Vos, M.C., van Leeuwen, W., van Belkum, A., Verbrugh, H.A. and Nouwen, J.L. (2005) The role of nasal carriage in Staphylococcus aureus infections. Lancet Infect. Dis., 5: 751-762. [Crossref]

3. Sangappa, S. and Thiagarajan, P. (2012) Methicillin resistant S. Aureus: Resistance gene and their regulation. Int. J. Pharm. Pharm. Sci., 4: 658-667.

4. Anon. (2001) European Antimicrobial Resistance Surveillance System (EARSS). Annual Report. National Institute of Public Health and the Environment, Bilthoven.

5. Rohrer, S., Bischoff, M., Rossi, J. and Berger-Bachi, B. (2003) Mechanisms of methicillin resistance. In: Fluit, A.C. and Schmitz, F.J., editors. MRSA: Current Perspectives. Caister Academic Press, Norfolk, England. p31-53.

6. Ito, T., Katayama, Y., Asada, K., Mori, N., Tsutsumimoto, K., Tiensasitorn, C. and Hiramatsu, K. (2001) Structural comparison of three types of staphylococcal cassette chromosome mec integrated in the chromosome in methicillin-resistant Staphylococcus aureus. Antimicrob. Agents Chemother., 45: 1323-1336. [Crossref] [PubMed] [PMC]

7. Ma, X.X., Ito, T., Tiensasitorn, C., Jamklang, M., Chongtrakool, P., Vavra, S.B., Daum, R.S. and Hiramatsu, K. (2002) Novel type of staphylococcal cassette chromosome mec identified in community-acquired methicillin resistant Staphylococcus aureus strains. Antimicrob. Agents Chemother., 46: 1147-1152. [Crossref] [PubMed] [PMC]

8. Kumar, R., Yadav, B.R. and Singh, R.S. (2011) Antibiotic resistance and pathogenicity factors in Staphylococcus aureus isolated from mastitic sahiwal cattle. J. Biosci., 36: 175-188. [Crossref] [PubMed]

9. Vishnupriya, S., Antony, P.X., Mukhopadhyay, H.K., Pillai, R.M., Thanislass, J., Srinivas, V.M. and Kumar, S.R. (2014) Methicillin resistant staphylococci associated with bovine mastitis and their zoonotic importance. Vet. World, 7: 422-427. [Crossref]

10. Chandrasekaran, D., Venkatesan, P., Tirumurugaan, K.G., Gowri, B., Subapriya, S. and Thirunavukkarasu, S. (2014) Sub-acute mastitis associated with methicillin resistant Staphylococcus aureus in a cow: A case report. J. Adv. Vet. Anim. Res., 1: 235-237. [Crossref]

11. Sharma, L., Verma, A.K., Kumar, A., Rahat, A., Neha . and Nigam, R. (2015) Incidence and pattern of antibiotic resistance of Staphylococcus aureus isolated from clinical and subclinical mastitis in cattle and buffaloes. Asia. J. Anim. Sci., 9: 100-109. [Crossref]

12. Agarwal, R.K., Bhilegaonkar, K.N., Singh, D.K., Kumar, A. and Rathore, R.S. (2003) In: Laboratory Manual for the Isolation and Identification of Food Borne Pathogens. 1st ed. IVRI, Izzatnagar. p38-39. [PubMed]

13. Monday, S.R. and Bohach, G.A. (1999) Use of multiplex PCR to detect classical and newly described pathogenic toxin gene in staphylococcal isolates. J. Clin. Microbiol., 37: 3411-3414. [PubMed] [PMC]

14. Braoios, A., Fluminhan, J.A. and Pizzolitto, A.C. (2009) Multiplex PCR use for Staphylococcus aureus identification and oxacillin and mupirocin resistance evaluation. J. Basic Appl. Pharm. Sci., 30: 303-307.

15. Kaushik, P., Anjay, Kumari, S., Bharti, S.K. and Dayal, S. (2014) Isolation and prevalence of Salmonella from chicken meat and cattle milk collected from local 30: Markets of Patna, India. Vet. World, 7: 62-65. [Crossref]

16. Wayne, P.A. (2002) Performance Standards of Antimicrobial Susceptibility: National Committee for Clinical Laboratory Standards (NCCLS). NCCLS Approved Standards. p100-159.

17. Garipcin, M. and Seker, E. (2015) Nasal carriage of methicillin-resistant Staphylococcus aureus in cattle and farm workers in Turkey. Vet. Arch., 85: 117-129.

18. Inegol, E. and Turkyilmaz, S. (2012) Determination of SCC mec types in methicillin resistant staphylococci isolated from cows and farm workers. Ankara. Univ. Vet. Fak. Derg., 59: 89-93. [Crossref]

19. Patchnee, P., Tadee, P., Arjkumpa, O., Love, D., Chanachai, K., Alter, T., Hinjoy, S. and Tharavichitkul, P. (2014) Occurrence and characterization of livestock associated methicillin resistant Staphylococcus aureus in pig industries of Northern Thailand. J. Vet. Sci., 15: 529-536. [Crossref] [PubMed] [PMC]

20. Mulligan, M.E., Murray, K.A., Standiford, H.C., John, J.F., Kauffmann, C.A. and Yu, V.L. (1993) Methicillin-resistant Staphylococcus aureus: A consensus review of the microbiology, pathogenesis, and epidemiology with implications for prevention and management. Ame. J. Med., 94: 313-328. [Crossref]

21. Pinho, M.G., Filipe, S.R., de Lencastre, H. and Tomasz, A. (2001) Complementation of the essential peptidoglycan trans-peptidase function of penicillin-binding protein 2 (PBP2) by the drug resistance protein PBP2A in Staphylococcus aureus. J. Bacteriol., 183: 6525-6531. [Crossref] [PubMed] [PMC]

22. Choi, S.M., Kim, S., Kim, C., Lee, D., Choi, J., Yoo, J., Kang, J., Shin, W. and Kang, W. (2003) Multiplex PCR for the detection of genes encoding aminoglycoside modifying enzymes and methicillin resistance among Staphylococcus species. J. Korean Med. Sci., 18: 631-636. [Crossref] [PubMed] [PMC]

23. Kalhor, H., Shariati, L., Validi, M., Tabatabaiefar, M.A. and Nafisi, M.R. (2012) Comparison of agar screen and duplex-PCR methods in determination of methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from nasal carriage. Afr. J. Microbiol. Res., 6: 3722-3726.

24. Alzohairy, M.A. (2011) Colonization and antibiotic susceptibility pattern of methicillin resistance Staphylococcus aureus (MRSA) among farm animals in Saudi Arabia. J. Bacteriol. Res., 3: 63-68.

25. Huber, H., Koller, S., Giezendanner, N., Stephan, R. and Zweifel, C. (2010) Prevalence and characteristics of methicillin-resistant Staphylococcus aureus in humans in contact with farm animals, in livestock, and in food of animal origin, Switzerland, 2009. Eur. Surv., 15: 7-10.

26. Lowy, F.D. (1998) Staphylococcus aureus infections. N. Engl. J. Med., 339: 520-532. [Crossref] [PubMed]

27. Peterson, G.K., Larsen, J., Harrison, E.M., Larsen, A.R., Morgan, F.J., Peacock, S.J., Parkhill, J., Zadoks, R.N. and Holmes, M.A. (2012) First detection of livestock-associated meticillin resistant Staphylococcus aureus CC398 in bulk tank milk in the United Kingdom. Eur. Surv., 17: 20337.

28. Assadullah, S., Kakru, D.K., Thoker, M.A., Bhat, F.A., Hussain, W. and Shah, A. (2003) Emergence of low level vancomycin resistance in MRSA. Indian J. Med. Microbiol., 21: 196-198. [PubMed]

29. Beninati, C., Reich, F., Muscolino, D., Giarratana, F., Panebianco, A., Klein, G. and Atanassova, V. (2015) ESBL-producing bacteria and MRSA isolated from poultry and Turkey products imported from Italy. Czech. J. Food Sci., 33: 97-102. [Crossref]