DOI QR코드

DOI QR Code

DNA microarray-based characterization and antimicrobial resistance phenotypes of clinical MRSA strains from animal hosts

  • Schmitt, Sarah (Institute for Food Safety and Hygiene, Vetsuisse Faculty University of Zurich) ;
  • Stephan, Roger (Institute for Food Safety and Hygiene, Vetsuisse Faculty University of Zurich) ;
  • Huebschke, Ella (Institute for Food Safety and Hygiene, Vetsuisse Faculty University of Zurich) ;
  • Schaefle, Daniel (Institute for Food Safety and Hygiene, Vetsuisse Faculty University of Zurich) ;
  • Merz, Axel (Institute for Food Safety and Hygiene, Vetsuisse Faculty University of Zurich) ;
  • Johler, Sophia (Institute for Food Safety and Hygiene, Vetsuisse Faculty University of Zurich)
  • Received : 2019.10.10
  • Accepted : 2020.05.11
  • Published : 2020.07.31

Abstract

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a leading cause of severe infections in humans and animals worldwide. Studies elucidating the population structure, staphylococcal cassette chromosome mec types, resistance phenotypes, and virulence gene profiles of animal-associated MRSA are needed to understand spread and transmission. Objectives: The objective of this study was to determine 1) clonal complexes and spa types, 2) resistance phenotypes, and 3) virulence/resistance gene profiles of MRSA isolated from animals in Switzerland. Methods: We analyzed 31 presumptive MRSA isolates collected from clinical infections in horses, dogs, cattle, sheep, and pigs, which had tested positive in the Staphaurex Latex Agglutination Test. The isolates were characterized by spa typing and DNA microarray profiling. In addition, we performed antimicrobial susceptibility testing using the VITEK 2 Compact system. Results: Characterization of the 31 presumptive MRSA isolates revealed 3 methicillinresistant Staphylococcus pseudintermedius isolates, which were able to grow on MRSA2 Brilliance agar. Of the 28 MRSA isolates, the majority was assigned to CC398 (86%), but CC8 (11%) and CC1 (4%) were also detected. The predominant spa type was t011 (n = 23), followed by t009 (n = 2), t034 (n = 1), t008 (n = 1), and t127 (n = 1). Conclusions: The results of this study extend the current body of knowledge on the population structure, resistance phenotypes, and virulence and resistance gene profiles of MRSA from livestock and companion animals.

Keywords

Acknowledgement

We thank all strain providers for their contribution.

References

  1. Robinson TP, Wertheim HF, Kakkar M, Kariuki S, Bu D, Price LB. Animal production and antimicrobial resistance in the clinic. Lancet. 2016;387(10014):e1-e3. https://doi.org/10.1016/S0140-6736(15)00730-8
  2. 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. https://doi.org/10.1111/j.1439-0450.1972.tb00439.x
  3. Ola Ojo M. Bacteriophage types and antibiotic sensitivity of Staphylococcus aureus isolated from swabs of the noses and skins of dogs. Vet Rec. 1972;91(6):152-153. https://doi.org/10.1136/vr.91.6.152
  4. Wendlandt S, Fessler AT, Monecke S, Ehricht R, Schwarz S, Kadlec K. The diversity of antimicrobial resistance genes among staphylococci of animal origin. Int J Med Microbiol. 2013;303(6-7):338-349. https://doi.org/10.1016/j.ijmm.2013.02.006
  5. Cuny C, Kock R, Witte W. Livestock associated MRSA (LA-MRSA) and its relevance for humans in Germany. Int J Med Microbiol. 2013;303(6-7):331-337. https://doi.org/10.1016/j.ijmm.2013.02.010
  6. McCarthy AJ, Lindsay JA, Loeffler A. Are all meticillin-resistant Staphylococcus aureus (MRSA) equal in all hosts? Epidemiological and genetic comparison between animal and human MRSA. Vet Dermatol. 2012;23(4):267-275, e53-e54. https://doi.org/10.1111/j.1365-3164.2012.01072.x
  7. Verkade E, Kluytmans J. Livestock-associated Staphylococcus aureus CC398: animal reservoirs and human infections. Infect Genet Evol. 2014;21:523-530. https://doi.org/10.1016/j.meegid.2013.02.013
  8. Fitzgerald JR. Livestock-associated Staphylococcus aureus: origin, evolution and public health threat. Trends Microbiol. 2012;20(4):192-198. https://doi.org/10.1016/j.tim.2012.01.006
  9. European Food Safety Authority; European Centre for Disease Prevention and Control. The European Union summary report on antimicrobial resistance in zoonotic and indicator bacteria from humans, animals and food in 2015. EFSA J. 2017;15(2):4694.
  10. Markey B, Leonard F, Archambault M, Cullinane A, Maguire D. Clinical veterinary microbiology. 2nd ed. St. Louis: Mosby Ltd.; 2013.
  11. Clinical and Laboratory Standards Institute. VET01S: Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals. 3rd ed. Wayne: CLSI; 2015.
  12. John MA, Burden J, Stuart JI, Reyes RC, Lannigan R, Milburn S, et al. Comparison of three phenotypic techniques for detection of methicillin resistance in Staphylococcus spp. reveals a species-dependent performance. J Antimicrob Chemother. 2009;63(3):493-496. https://doi.org/10.1093/jac/dkn527
  13. Livermore DM, Struelens M, Amorim J, Baquero F, Bille J, Canton R, et al. Multicentre evaluation of the VITEK 2 Advanced Expert System for interpretive reading of antimicrobial resistance tests. J Antimicrob Chemother. 2002;49(2):289-300. https://doi.org/10.1093/jac/49.2.289
  14. Monecke S, Slickers P, Ehricht R. Assignment of Staphylococcus aureus isolates to clonal complexes based on microarray analysis and pattern recognition. FEMS Immunol Med Microbiol. 2008;53(2):237-251. https://doi.org/10.1111/j.1574-695X.2008.00426.x
  15. Wattinger L, Stephan R, Layer F, Johler S. Comparison of Staphylococcus aureus isolates associated with food intoxication with isolates from human nasal carriers and human infections. Eur J Clin Microbiol Infect Dis. 2012;31(4):455-464. https://doi.org/10.1007/s10096-011-1330-y
  16. Johler S, Layer F, Stephan R. Comparison of virulence and antibiotic resistance genes of food poisoning outbreak isolates of Staphylococcus aureus with isolates obtained from bovine mastitis milk and pig carcasses. J Food Prot. 2011;74(11):1852-1859. https://doi.org/10.4315/0362-028X.JFP-11-192
  17. Huber H, Koller S, Giezendanner N, Stephan R, Zweifel C. Prevalence and characteristics of meticillin-resistant Staphylococcus aureus in humans in contact with farm animals, in livestock, and in food of animal origin, Switzerland, 2009. Euro Surveill. 2010;15(16):19542.
  18. Overesch G, Buttner S, Rossano A, Perreten V. The increase of methicillin-resistant Staphylococcus aureus (MRSA) and the presence of an unusual sequence type ST49 in slaughter pigs in Switzerland. BMC Vet Res. 2011;7:30. https://doi.org/10.1186/1746-6148-7-30
  19. Price LB, Stegger M, Hasman H, Aziz M, Larsen J, Andersen PS, et al. Staphylococcus aureus CC398: host adaptation and emergence of methicillin resistance in livestock. mBio. 2012;3(1):e00305-11.
  20. Schijffelen MJ, Boel CH, van Strijp JA, Fluit AC. Whole genome analysis of a livestock-associated methicillin-resistant Staphylococcus aureus ST398 isolate from a case of human endocarditis. BMC Genomics. 2010;11:376. https://doi.org/10.1186/1471-2164-11-376
  21. Heikinheimo A, Johler S, Karvonen L, Julmi J, Fredriksson-Ahomaa M, Stephan R. New dominant spa type t2741 in livestock-associated MRSA (CC398-MRSA-V) in Finnish fattening pigs at slaughter. Antimicrob Resist Infect Control. 2016;5:6. https://doi.org/10.1186/s13756-016-0105-8
  22. Loncaric I, Kunzel F, Licka T, Simhofer H, Spergser J, Rosengarten R. Identification and characterization of methicillin-resistant Staphylococcus aureus (MRSA) from Austrian companion animals and horses. Vet Microbiol. 2014;168(2-4):381-387. https://doi.org/10.1016/j.vetmic.2013.11.022
  23. Cuny C, Abdelbary MM, Kock R, Layer F, Scheidemann W, Werner G, et al. Methicillin-resistant Staphylococcus aureus from infections in horses in Germany are frequent colonizers of veterinarians but rare among MRSA from infections in humans. One Health. 2015;2:11-17. https://doi.org/10.1016/j.onehlt.2015.11.004
  24. Vincze S, Stamm I, Kopp PA, Hermes J, Adlhoch C, Semmler T, et al. Alarming proportions of methicillin-resistant Staphylococcus aureus (MRSA) in wound samples from companion animals, Germany 2010-2012. PLoS One. 2014;9(1):e85656. https://doi.org/10.1371/journal.pone.0085656
  25. European Food Safety Authority; European Centre for Disease Prevention and Control. EU Summary Report on antimicrobial resistance in zoonotic and indicator bacteria from humans, animals and food in 2013. EFSA J. 2015;13(2):4036.
  26. Albrecht N, Jatzwauk L, Slickers P, Ehricht R, Monecke S. Clonal replacement of epidemic methicillin-resistant Staphylococcus aureus strains in a German university hospital over a period of eleven years. PLoS One. 2011;6(11):e28189. https://doi.org/10.1371/journal.pone.0028189
  27. Cuny C, Strommenger B, Witte W, Stanek C. Clusters of infections in horses with MRSA ST1, ST254, and ST398 in a veterinary hospital. Microb Drug Resist. 2008;14(4):307-310. https://doi.org/10.1089/mdr.2008.0845
  28. Huber H, Giezendanner N, Stephan R, Zweifel C. Genotypes, antibiotic resistance profiles and microarray-based characterization of methicillin-resistant Staphylococcus aureus strains isolated from livestock and veterinarians in Switzerland. Zoonoses Public Health. 2011;58(5):343-349. https://doi.org/10.1111/j.1863-2378.2010.01353.x
  29. Franco A, Hasman H, Iurescia M, Lorenzetti R, Stegger M, Pantosti A, et al. Molecular characterization of spa type t127, sequence type 1 methicillin-resistant Staphylococcus aureus from pigs. J Antimicrob Chemother. 2011;66(6):1231-1235. https://doi.org/10.1093/jac/dkr115
  30. Heffernan H, Bakker S. Annual survey of methicillin-resistant Staphylococcus aureus (MRSA), 2015. Wellington: ESR; 2016.
  31. Wipf JR, Perreten V. Methicillin-resistant Staphylococcus aureus isolated from dogs and cats in Switzerland. Schweiz Arch Tierheilkd. 2016;158(6):443-450. https://doi.org/10.17236/sat00070
  32. Argudin MA, Tenhagen BA, Fetsch A, Sachsenroder J, Kasbohrer A, Schroeter A, et al. Virulence and resistance determinants of German Staphylococcus aureus ST398 isolates from nonhuman sources. Appl Environ Microbiol. 2011;77(9):3052-3060. https://doi.org/10.1128/AEM.02260-10
  33. Schwarz S, Silley P, Simjee S, Woodford N, van Duijkeren E, Johnson AP, et al. Editorial: assessing the antimicrobial susceptibility of bacteria obtained from animals. J Antimicrob Chemother. 2010;65(4):601-604. https://doi.org/10.1093/jac/dkq037
  34. Pottumarthy S, Schapiro JM, Prentice JL, Houze YB, Swanzy SR, Fang FC, et al. Clinical isolates of Staphylococcus intermedius masquerading as methicillin-resistant Staphylococcus aureus. J Clin Microbiol. 2004;42(12):5881-5884. https://doi.org/10.1128/JCM.42.12.5881-5884.2004
  35. Wu MT, Burnham CA, Westblade LF, Dien Bard J, Lawhon SD, Wallace MA, et al. Evaluation of oxacillin and cefoxitin disk and MIC breakpoints for prediction of methicillin resistance in human and veterinary isolates of Staphylococcus intermedius group. J Clin Microbiol. 2016;54(3):535-542. https://doi.org/10.1128/JCM.02864-15
  36. van Duijkeren E, Hengeveld PD, Albers M, Pluister G, Jacobs P, Heres L, et al. Prevalence of methicillin-resistant Staphylococcus aureus carrying mecA or mecC in dairy cattle. Vet Microbiol. 2014;171(3-4):364-367. https://doi.org/10.1016/j.vetmic.2013.12.024
  37. Bannoehr J, Guardabassi L. Staphylococcus pseudintermedius in the dog: taxonomy, diagnostics, ecology, epidemiology and pathogenicity. Vet Dermatol. 2012;23(4):253-266, e51-e52. https://doi.org/10.1111/j.1365-3164.2012.01046.x
  38. Fetsch A, Johler S. Staphylococcus aureus as a foodborne pathogen. Curr Clin Microbiol Rep. 2018;5(2):88-96. https://doi.org/10.1007/s40588-018-0094-x
  39. Kraushaar B, Ballhausen B, Leeser D, Tenhagen BA, Kasbohrer A, Fetsch A. Antimicrobial resistances and virulence markers in Methicillin-resistant Staphylococcus aureus from broiler and turkey: a molecular view from farm to fork. Vet Microbiol. 2017;200:25-32. https://doi.org/10.1016/j.vetmic.2016.05.022
  40. Kittl S, Brodard I, Heim D, Andina-Pfister P, Overesch G. Methicillin-resistant Staphyococcus aureus strains in Swiss pigs and their relation to isolates from farmers and veterinarians. Appl Environ Microbiol. 2020;86(5):e01865-19.