Investigation of Antibiotic Resistance Pattern and Detection of Methicillin-Resistant Strains (MRSA) in Staphylococcus Aureus Isolates Associated with Bovine Mastitis

Document Type : Original Article(s)

Authors

1 Associate Professor, Department of Microbiology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

2 Department of Microbiology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

3 MSc Student, Department of Microbiology, School of Medicine AND Student Research Committee, Isfahan University of Medical Sciences, Isfahan, Iran

Abstract

Background: Staphylococcus aureus is one of the main causative agents in bovine mastitis. Resistance of some Staphylococcus aureus strains to antibiotics, particularly methicillin, can complicate the treatment of its infections. Moreover, the transmission of resistant strains into human is also important. Thus, the aim of this study was to investigate the prevalence of mastitis caused by Staphylococcus aureus in Isfahan dairy herds, Iran, and to determine the antimicrobial susceptibility pattern of the isolates.Methods: A total of 450 milk samples were collected. After inoculation and applying the relevant tests, antibiotic resistance was determined via disk diffusion method in isolates. D-test method was performed for the detection of inducible clindamycin resistance (ICR). Then, methicillin-resistant strains (MRSA) were detected using oxacillin agar screening and cefoxitin disc diffusion methods.Findings: Fifty-four Staphylococcus aureus were isolated. All isolates were sensitive to ciprofloxacin and gentamicin. Furthermore, 31, 25, 8, 7 and 2 isolates were resistant to erythromycin, clindamycin, tetracycline, oxacillin and cotrimoxazole, respectively. D-test was negative for all isolates. Nine methicillin-resistant strains were identified using oxacillin agar screening and ten methicillin-resistant strains were detected using cefoxitin disc diffusion methods.Conclusion: Since it is expected that the prevalence of Staphylococcus aureus, especially antibiotic resistant strains in herds, be too negligible, even the low incidence reported in this study can lead to many problems. Therefore, applying an appropriate antimicrobial susceptibility test before the beginning of antibiotic therapy in bovine mastitis cases is of primary importance.

Keywords


  1. Philpot WN, Nickerson SC. Winning the fight against mastitis. Naperville, IL: Westfalia Surge; 2000.
  2. Saleki K, Moradi H. Bacterial agents of mastitis in dairy cow farms in Ilam city. J Ilam Univ Med Sci 2013; 20(4): 88-95. [In Persian].
  3. Barlow J. Mastitis therapy and antimicrobial susceptibility: a multispecies review with a focus on antibiotic treatment of mastitis in dairy cattle. J Mammary Gland Biol Neoplasia 2011; 16(4): 383-407.
  4. Schroeder JW. Bovine mastitis and milking management [Online]. [cited 2010]; Available from:URL:http://www.ag.ndsu.edu/pubs/ansci/dairy/as1129.pdf
  5. Barker AR, Schrick FN, Lewis MJ, Dowlen HH, Oliver SP. Influence of clinical mastitis during early lactation on reproductive performance of Jersey cows. J Dairy Sci 1998; 81(5): 1285-90.
  6. Risco CA, Donovan GA, Hernandez J. Clinical mastitis associated with abortion in dairy cows. J Dairy Sci 1999; 82(8): 1684-9.
  7. Hashemi M, Kafi M, Safdarian M. The prevalence of clinical and subclinical mastitis in dairy cows in the central region of Fars province, south of Iran. Iran J Vet Res 2011; 12(3): 236-41.
  8. Nava H, Tajik P. Evaluation of two screening test, CMT and ECT to detect subclinical mastitis in dairy cow. J Fac Vet Med Univ Tehran 2002; 57(4):91-6.
  9. Holmes MA, Zadoks RN. Methicillin resistant S. aureus in human and bovine mastitis. J Mammary Gland Biol Neoplasia 2011; 16(4): 373-82.
  10. Kazemi J, Ahmadi M, Dastmalchi Saei H, Adib Hesami M. The effect of silver nanoparticles and beta-lactam antibiotics individually and in combination on Staphylococcus aureus isolated from cattle mastitis. Vet Microbiol 2012; 8(1): 57-66.
  11. Atyabi N, Vodjgani M, Gharagozloo F, Bahonar A. Prevalence of bacterial mastitis in cattle from the farms around Tehran. Iran J Vet Res 2006; 7(3): 76-9.
  12. Ghorbanpoor M, Seyfiabad Shapouri M, Moatamedi H, Jamshidian M, Gooraninejad S. Comparison of PCR and bacterial culture methods for diagnosis of dairy cattle's subclinical mastitis caused by Staphylococcus aureus. J Vet Res 2007; 62(4): 87-91.
  13. Vanderhaeghen W, Cerpentier T, Adriaensen C, Vicca J, Hermans K, Butaye P. Methicillin-resistant Staphylococcus aureus (MRSA) ST398 associated with clinical and subclinical mastitis in Belgian cows. Vet Microbiol 2010; 144(1-2): 166-71.
  14. Devriese LA, Hommez J. Epidemiology of methicillin-resistant Staphylococcus aureus in dairy herds. Res Vet Sci 1975; 19(1): 23-7.
  15. Devriese LA, Van Damme LR, Fameree L. Methicillin (cloxacillin)-resistant Staphylococcus aureus strains isolated from bovine mastitis cases. Zentralbl Veterinarmed B 1972; 19(7): 598-605.
  16. Mohammad Sadegh M, Askari Badouei M, Gorjidooz M, Daneshvar M and Koochakzadeh A. A study on the clinical Coliform mastitis of Holstein cows on Garmsar suburban dairy farms. Vet Microbiol 2012; 8(2): 137-49.
  17. Abera M, Demie B, Aragaw K, Regassa F, Regassa A. Isolation and identification of Staphylococcus aureus from bovine mastitic milk and their drug resistance patterns in Adama town, Ethiopia. J Vet Med Anim Health 2010; 2(3): 29-34.
  18. Vojgani M, Gharagozloo F, Bahonar A, Darabi M, Jafari H. Evaluation of therapeutic effects of topical application of eucalyptus essential oil (essence) on experimental mastitis caused by Streptococcus agalactiae. Iran Vet J 2006; 10(12): 5-14. [In Persian].
  19. Ghorbanpoor M, Seifiabad Shapouri M, Moatamedi H, Jamshidian M, Gooraninejad S. Comparison of microbial culture, PCR and CMT for diagnosis of dairy cattle's staphylococcal and streptococcal subclinical mastitis. Iran Vet J 2007; 3(1): 63-70.
  20. Ebrahimi A, Akhavan Taheri M. Characteristics of staphylococci isolated from clinical and subclinical mastitis cows in Shahrekord. Iran J Vet Res 2009; 10(3): 273-7.
  21. Clinical and Laboratory Standards Institute. Performance Standards for Antimicrobial Disk Susceptibility Tests; Approved Standard (M02-A11.). 11th ed [Online]. [cited 2012]; Available from: URL:
  22. http://antimicrobianos.com.ar/ATB/wp-content/ uploads/2012/11/01-CLSI-M02-A11-2012.pdf.
  23. Sedighi I, Rasoul Yousefi M, Pak N, Seif Rabiee MA. D-test method for detection of inducible clindamycin resistance in staphylococcus aureus. Iran J Pediatr 2009; 19(3): 293-7.
  24. Ghasemian Safai H, Rahimi E. Detection of classical enterotoxins of Staphylococcus aureus strains isolated from bovine subclinical mastitis in Isfahan, Iran. Vet Microbiol 2010; 141(3-4): 393–4.
  25. Anand KB, Agrawal P, Kumar S, Kapila K. Comparison of cefoxitin disc diffusion test, oxacillin screen agar, and PCR for mecA gene for detection of MRSA. Indian J Med Microbiol 2009; 27(1): 27-9.
  26. Rezazadeh M, Yousefi Mashouf R, Sarmadyan H, Ghaznavi-Rad E. Antibiotic profile of methicillin-resistant Staphylococcus aureus with multiple-drug resistances isolated from nosocomial infections in Vali-Asr Hospital of Arak. J Arak Univ Med Sci 2013; 16(71): 29-37. [In Persian].
  27. Caierao J, Superti S, Dias CA, d'Azevedo PA. Automated systems in the identification and determination of methicillin resistance among coagulase negative staphylococci. Mem Inst Oswaldo Cruz 2006; 101(3): 277-80.
  28. Khan Nazer A. H, Sarmadi R. Prevalence of clinical and subclinical mastitis, antibiotic resistance and determination of minimum inhibitory Concentration (MIC) in Staphylococcus aureus and Escherichia coli isolated from cases of bovine Mastitis. J Vet Res 2005; 60(3): 247-52.
  29. Busato A, Trachsel P, Schallibaum M, Blum JW. Udder health and risk factors for subclinical mastitis in organic dairy farms in Switzerland. Prev Vet Med 2000; 44(3-4): 205-20.
  30. Phuektes P, Mansell PD, Browning GF. Multiplex polymerase chain reaction assay for simultaneous detection of Staphylococcus aureus and streptococcal causes of bovine mastitis. J Dairy Sci 2001; 84(5): 1140-8.
  31. Turkyilmaz S, Tekbiyik S, Oryasin E, Bozdogan B. Molecular epidemiology and antimicrobial resistance mechanisms of methicillin-resistant Staphylococcus aureus isolated from bovine milk. Zoonoses Public Health 2010; 57(3): 197-203.
  32. Skov R, Smyth R, Clausen M, Larsen AR, Frimodt-Moller N, Olsson-Liljequist B, et al. Evaluation of a cefoxitin 30 microg disc on Iso-Sensitest agar for detection of methicillin-resistant Staphylococcus aureus. J Antimicrob Chemother 2003; 52(2): 204-7.
  33. Shariati L, Validi M, Tabatabaiefar MA, Karimi A, Nafisi MR. Comparison of real-time PCR with disk diffusion, agar screen and E-test methods for detection of methicillin-resistant Staphylococcus aureus. Curr Microbiol 2010; 61(6): 520-4.