J Vet Sci.  2012 Jun;13(2):153-161. 10.4142/jvs.2012.13.2.153.

In vitro susceptibility of Staphylococcus aureus strains isolated from cows with subclinical mastitis to different antimicrobial agents

• Behiry AE
• Schlenker
• Szabo I
• Roesler U

• KMID: 1376189
• DOI: http://doi.org/10.4142/jvs.2012.13.2.153

Abstract

Sensitivity to commercial teat dips (nonoxinol-9 iodine complex and chlorhexidine digluconate) of 56 Staphylococcus (S.) aureus strains isolated from quarter milk samples of various German dairy herds treated with different teat dipping schemes was investigated in this study. The minimum inhibitory concentration was determined using a broth macrodilution method according to the German Veterinary Association guidelines. The main objective of the current study was to induce in vitro resistance induction of S. aureus to chemical disinfectants. Ten different strains were repeatedly passed ten times in growth media with sub-lethal concentrations of disinfectants. Nine strains showed a significant reduction in susceptibility to the nonoxinol-9 iodine complex but only one strain developed resistance to chlorhexidine digluconate. Stability of the acquired resistance was observed in all S. aureus strains adapted to the nonoxinol-9 iodine complex and chlorhexidine digluconate. In contrast, simultaneous resistance to different antibiotics was not observed in any of the ten investigated S. aureus strains. However, the isolates exhibited a high degree of resistance to penicillin G. Based on these results, resistance of S. aureus to chemical disinfectants may be more likely to develop if the chemicals are used at concentrations lower than that required for an optimal biocidal effect.

Keyword

antibiotics; bovine mastitis; resistance induction; Staphylococcus aureus; teat dips

MeSH Terms

Animals
Anti-Bacterial Agents/pharmacology
Cattle
Chlorhexidine/*pharmacology
Disinfectants/pharmacology
Dose-Response Relationship, Drug
Drug Resistance, Bacterial
Female
Germany/epidemiology
Iodine/chemistry/*pharmacology
Mastitis, Bovine/epidemiology/*microbiology
Nonoxynol/*pharmacology
Staphylococcal Infections/epidemiology/microbiology/*veterinary
Staphylococcus aureus/classification/*drug effects

Reference

1. Aarestrup FM, Jensen NE. Development of penicillin resistance among Staphylococcus aureus isolated from bovine mastitis in Denmark and other countries. Microb Drug Resist. 1998. 4:247–256.

2. Birosová L, Mikulášová M. Development of triclosan and antibiotic resistance in Salmonella enterica serovar Typhimurium. J Med Microbiol. 2009. 58:436–441.

3. Capurro A. Diagnostic and epidemiological studies of staphylococci in bovine mastitis. 2009. Uppsala: Swedish University of Agricultural Sciences;Doctoral dissertation.

4. Clinical and Laboratory Standards Institute. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically; Approved Standard. 2009. 8th ed. Wayne: Clinical and Laboratory Standards Institute;1–65.

5. Compton CWR, Heuer C, Parker K, McDougall S. Risk factors for peripartum mastitis in pasture-grazed dairy heifers. J Dairy Sci. 2007. 90:4171–4180.

6. Cottell A, Denyer SP, Hanlon GW, Ochs D, Maillard JY. Triclosan-tolerant bacteria: changes in susceptibility to antibiotics. J Hosp Infect. 2009. 72:71–76.

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

8. Escalada MG, Harwood JL, Maillard JY, Ochs D. Triclosan inhibition of fatty acid synthesis and its effect on growth of Escherichia coli and Pseudomonas aeruginosa. J Antimicrob Chemother. 2005. 55:879–882.

9. Gentilini E, Denamiel G, Llorente P, Godaly S, Rebuelto M, DeGregorio O. Antimicrobial susceptibility of Staphylococcus aureus isolated from bovine mastitis in Argentina. J Dairy Sci. 2000. 83:1224–1227.

10. Ghorbanpoor M, Seyfi Abad Shapouri MR, 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. 62:87–91.

11. Gilbert P, Allison DG, McBain AJ. Biofilms in vitro and in vivo: do singular mechanisms imply cross-resistance? J Appl Microbiol. 2002. 92:Suppl. 98S–110S.

12. Gruet P, Maincent P, Berthelot X, Kaltsatos V. Bovine mastitis and intramammary drug delivery: review and perspectives. Adv Drug Deliv Rev. 2001. 50:245–259.

13. Hogan JS, Smith KL. Prolonged in vitro exposure of Staphylococcus aureus to germicidal teat dips. J Dairy Sci. 1989. 72:1052–1056.

14. Huet AA, Raygada JL, Mendiratta K, Seo SM, Kaatz GW. Multidrug efflux pump overexpression in Staphylococcus aureus after single and multiple in vitro exposures to biocides and dyes. Microbiology. 2008. 154:3144–3153.

15. Jones A, Higgs TM, Neave FK, Smith A. The sensitivity of bovine staphylococci, streptococci and corynebacteria to cloxacillin and various other antibiotics. J Dairy Res. 1967. 34:249–255.

16. Jurgens DJ, Sattar SA, Mah TF. Chloraminated drinking water does not generate bacterial resistance to antibiotics in Pseudomonas aeruginosa biofilms. Lett Appl Microbiol. 2008. 46:562–567.

17. Karatzas KA, Webber MA, Jorgensen F, Woodward MJ, Piddock LJ, Humphrey TJ. Prolonged treatment of Salmonella enterica serovar Typhimurium with commercial disinfectants selects for multiple antibiotic resistance, increased efflux and reduced invasiveness. J Antimicrob Chemother. 2007. 60:947–955.

18. Martineau F, Picard FJ, Lansac N, Ménard C, Roy PH, Ouellette M, Bergeron MG. Correlation between the resistance genotype determined by multiplex PCR assays and the antibiotic susceptibility patterns of Staphylococcus aureus and Staphylococcus epidermidis. Antimicrob Agents Chemother. 2000. 44:231–238.

19. McDonnell G, Russell AD. Antiseptics and disinfectants: activity, action, and resistance. Clin Microbiol Rev. 1999. 12:147–179.

20. National Mastitis Council. Summary of Peer-Reviewed Publications on Efficacy of Premilking and Postmilking Teat Disinfectants Published since 1980. 2009. Verona: National Mastitis Council;213–227.

21. Nunes SF, Bexiga R, Cavaco LM, Vilela CL. Technical note: antimicrobial susceptibility of Portuguese isolates of Staphylococcus aureus and Staphylococcus epidermidis in subclinical bovine mastitis. J Dairy Sci. 2007. 90:3242–3246.

22. Oliver SP, King SH, Lewis MJ, Torre PM, Matthews KR, Dowlen HH. Efficacy of chlorhexidine as a postmilking teat disinfectant for the prevention of bovine mastitis during lactation. J Dairy Sci. 1990. 73:2230–2235.

23. Piddock LJV. Clinically relevant chromosomally encoded multidrug resistance efflux pumps in bacteria. Clin Microbiol Rev. 2006. 19:382–402.

24. Pitkälä A, Haveri M, Pyörälä S, Myllys V, Honkanen-Buzalski T. Bovine mastitis in Finland 2001--prevalence, distribution of bacteria, and antimicrobial resistance. J Dairy Sci. 2004. 87:2433–2441.

25. Prince HN, Nonemaker WS, Norgard RC, Prince DL. Drug resistance studies with topical antiseptics. J Pharm Sci. 1978. 67:1629–1631.

26. Pyörälä S, Taponen S. Coagulase-negative staphylococci-emerging mastitis pathogens. Vet Microbiol. 2009. 134:3–8.

27. Randall LP, Cooles SW, Coldham NG, Penuela EG, Mott AC, Woodward MJ, Piddock LJ, Webber MA. Commonly used farm disinfectants can select for mutant Salmonella enterica serovar Typhimurium with decreased susceptibility to biocides and antibiotics without compromising virulence. J Antimicrob Chemother. 2007. 60:1273–1280.

28. Rogers CK. Non-prescription Drugs Advisory Committee. FDA reviewer's literature summary regarding potential environmental hazards of biocides. Consumer Antiseptic Drug Products Review. 2005. Silver Spring: Food and Drug Administration;1–11.

29. Russell AD, Tattawasart U, Maillard JY, Furr JR. Possible link between bacterial resistance and use of antibiotics and biocides. Antimicrob Agents Chemother. 1998. 42:2151.

30. Saei HD, Ahmadi M, Mardani K, Batavani RA. Genotyping of Staphylococcus aureus isolated from bovine mastitis based on PCR-RFLP analysis of the aroA gene. Comp Clin Pathol. 2010. 19:163–168.

31. Sauer S, Freiwald A, Maier T, Kube M, Reinhardt R, Kostrzewa M, Geider K. Classification and identification of bacteria by mass spectrometry and computational analysis. PLoS One. 2008. 3:e2843.

32. Scientific Committee on Emerging and Newly Identified Health Risk. Assessment of the Antibiotic Resistance Effects of Biocides. 2009. Brussels: Scientific Committee on Emerging and Newly Identified Health Risk;1–87.

33. Sears PM, McCarthy KK. Management and treatment of staphylococcal mastitis. Vet Clin North Am Food Anim Pract. 2003. 19:171–185.

34. Shi D, Hao Y, Zhang A, Wulan B, Fan X. Antimicrobial resistance of Staphylococcus aureus isolated from bovine mastitis in China. Transbound Emerg Dis. 2010. 57:221–224.

35. Sutra L, Poutrel B. Virulence factors involved in the pathogenesis of bovine intramammary infections due to Staphylococcus aureus. J Med Microbiol. 1994. 40:79–89.

36. Szumała A, Pernak J. The natural and laboratory resistance of gram-negative rods to quaternary ammonium chlorides with alkylthiomethyl radical. Pharmazie. 1986. 41:521.

37. Watts JL, Salmon SA. Activity of selected antimicrobial agents against strains of Staphylococcus aureus isolated from bovine intramammary infections that produce β-lactamase. J Dairy Sci. 1997. 80:788–791.

38. Werckenthin C, Cardoso M, Martel JL, Schwarz S. Antimicrobial resistance in staphylococci from animals with particular reference to bovine Staphylococcus aureus, porcine Staphylococcus hyicus, and canine Staphylococcus intermedius. Vet Res. 2001. 32:341–362.

39. Whist AC, Østerås O, Sølverød L. Staphylococcus aureus and Streptococcus dysgalactiae in Norwegian herds after introduction of selective dry cow therapy and teat dipping. J Dairy Res. 2007. 74:1–8.

40. Yilmaz A, Kaleta EF. Suitability of two commercial preparations for disinfections against methicillin-resistant Staphylococcus aureus in veterinary medicine. Dtsch Tierarztl Wochenschr. 2009. 116:180–185.