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Identification and methicillin resistance of coagulase-negative staphylococci isolated from nasal cavity of healthy horses

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Abstract

The aim of this study was an analysis of the staphylococcal flora of the nasal cavity of 42 healthy horses from 4 farms, along with species identification of CoNS isolates and determination of resistance to 18 antimicrobial agents, particularly phenotypic and genotypic methicillin resistance. From the 81 swabs, 87 staphylococci were isolated. All isolates possessed the gap gene but the coa gene was not detected in any of these isolates. Using PCR-RFLP of the gap gene, 82.8% of CoNS were identified: S. equorum (14.9%), S. warneri (14.9%), S. sciuri (12.6%), S. vitulinus (12.6%), S. xylosus (11.5%), S. felis (5.7%), S. haemolyticus (3.4%), S. simulans (3.4%), S. capitis (1.1%), S. chromogenes (1.1%), and S. cohnii subsp. urealyticus (1.1%). To our knowledge, this was the first isolation of S. felis from a horse. The species identity of the remaining Staphylococcus spp. isolates (17.2%) could not be determined from the gap gene PCR-RFLP analysis and 16S rRNA gene sequencing data. Based on 16S–23S intergenic transcribed spacer PCR, 11 different ITS-PCR profiles were identified for the 87 analyzed isolates. Results of API Staph were consistent with molecular identification of 17 (19.5%) isolates. Resistance was detected to only 1 or 2 of the 18 antimicrobial agents tested in the 17.2% CoNS isolates, including 6.9% MRCoNS. The mecA gene was detected in each of the 5 (5.7%) phenotypically cefoxitin-resistant isolates and in 12 (13.8%) isolates susceptible to cefoxitin. In total, from 12 horses (28.6%), 17 (19.5%) MRCoNS were isolated. The highest percentage of MRCoNS was noted among S. sciuri isolates (100%).

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References

  • Bagcigil, F.A., Moodley, A., Baptiste, K.E., Jensen, V.F., and Guardabassi, L. 2007. Occurrence, species distribution, antimicrobial resistance and clonality of methicillin- and erythromycin-resistant staphylococci in the nasal cavity of domestic animals. Vet. Microbiol. 121, 307–315.

    Article  PubMed  CAS  Google Scholar 

  • Baptiste, K.E., Williams, K., Willams, N.J., Wattret, A., Clegg, P.D., Dawson, S., Corkill, J.E., O’Neill, T., and Hart, C.A. 2005. Methicillin-resistant staphylococci in companion animals. Emerg. Infect. Dis. 11, 1942–1944.

    Article  PubMed  Google Scholar 

  • Becker, K., Harmsen, D., Mellmann, A., Meier, Ch., Schumann, P., Peters, G., and von Eiff, Ch. 2004. Development and evaluation of a quality-controlled ribosomal sequence database for 16S ribosomal DNA-based identification of Staphylococcus species. J. Clin. Microbiol. 42, 4988–4995.

    Article  PubMed  CAS  Google Scholar 

  • Burton, S., Reid-Smith, R., McClure, J.T., and Weese, J.S. 2008. Staphylococcus aureus colonization in healthy horses in Atlantic Canada. Can. Vet. J. 49, 797–799.

    PubMed  Google Scholar 

  • Busscher, J.F., van Duijkeren, E., and Sloet van Oldruitenborgh-Oosterbaan, M.M. 2006. The prevalence of methicillin-resistant staphylococci in healthy horses in the Netherlands. Vet. Microbiol. 113, 131–136.

    Article  PubMed  CAS  Google Scholar 

  • CLSI. 2008. Performance standards for antimicrobial susceptibility testing; 18th informational supplement. CLSI document M100-S18. Clinical and Laboratory Standards Institute, Wayne, PA, USA.

    Google Scholar 

  • Couto, I., Pereira, S., Miragaia, M., Sanches Santos, I., and de Lencastre, H. 2001. Identification of clinical staphylococcal isolates from humans by internal transcribed spacer PCR. J. Clin. Microbiol. 39, 3099–3103.

    Article  PubMed  CAS  Google Scholar 

  • da Silva, E. and da Silva, N. 2005. Coagulase gene typing of Staphylococcus aureus isolated from cows with mastitis in southeastern Brazil. Can. J. Vet. Res. 69, 260–264.

    PubMed  Google Scholar 

  • De Martino, L., Lucido, M., Mallardo, K., Facello, B., Mallardo, M., Iovane, G., Pagnini, U., Tufano, M.A., and Catalanotti, P. 2010. Methicillin-resistant staphylococci isolated from healthy horses and horse personnel in Italy. J. Vet. Diagn. Invest. 22, 77–82.

    Article  PubMed  Google Scholar 

  • Fiebelkorn, K.R., Crawford, S.A., McElmeel, M.L., and Jorgensen, J.H. 2003. Practical disc diffusion method for detection of inducible clindamycin resistance in Staphylococcus aureus and coagulase-negative staphylococci. J. Clin. Microbiol. 41, 4740–4744.

    Article  PubMed  CAS  Google Scholar 

  • Fuda, C., Suvorov, M., Shi, Q., Hesek, D., Lee, M., and Mobashery, S. 2007. Shared functional attributes between the mecA gene product of Staphylococcus sciuri and penicillin-binding protein 2a of methicillin-resistant Staphylococcus aureus. Biochemistry 46, 8050–8057.

    Article  PubMed  CAS  Google Scholar 

  • Fujita, S., Senda, Y., Iwagami, T., and Hashimoto, T. 2005. Rapid identification of staphylococcal strains from positive-testing blood culture bottles by internal transcribed spacer PCR followed by microchip gel electrophoresis. J. Clin. Microbiol. 43, 1149–1157.

    Article  PubMed  CAS  Google Scholar 

  • Ghebremedhin, B., Layer, F., König, W., and König, B. 2008. Genetic classification and distinguishing of Staphylococcus species based on different partial gap, 16S rRNA, hsp60, rpoB, sodA, and tuf gene sequences. J. Clin. Microbiol. 46, 1019–1025.

    Article  PubMed  CAS  Google Scholar 

  • Gürtler, V. and Stanisich, V.A. 1996. New approaches to typing and identification of bacteria using the 16S–23S rDNA spacer region. Microbiology 142, 3–16.

    Article  PubMed  Google Scholar 

  • Harmsen, D. and Karch, H. 2004. 16S rDNA for diagnosing pathogens: a living tree. ASM News 70, 19–24.

    Google Scholar 

  • Harmsen, D., Rothgänger, J., Frosch, M., and Albert, J. 2002. RIDOM: ribosomal differentiation of medical micro-organisms database. Nucleic Acids Res. 30, 416–417.

    Article  PubMed  CAS  Google Scholar 

  • Heikens, E., Fleer, A., Paauw, A., Florijn, A., and Fluit, A.C. 2005. Comparison of genotypic and phenotypic methods for species-level identification of clinical isolates of coagulase-negative staphylococci. J. Clin. Microbiol. 43, 2286–2290.

    Article  PubMed  CAS  Google Scholar 

  • Higuchi, W., Takano, T., Teng, L.J., and Yamamoto, T. 2008. Structure and specific detection of staphylococcal cassette chromosome mec type VII. Biochem. Biophys. Res. Commun. 377, 752–756.

    Article  PubMed  CAS  Google Scholar 

  • Huber, H., Ziegler, D., Pflüger, V., Vogel, G., Zweifel, C., and Stephan, R. 2011. Prevalence and characteristics of methicillin-resistant coagulase-negative staphylococci from livestock, chicken carcasses, bulk tank milk, minced meat, and contact persons. BMC Vet. Res. 7, 6.

    Article  PubMed  Google Scholar 

  • Igimi, S., Kawamura, S., Takahashi, E., and Mitsuoka, T. 1989. Staphylococcus felis, a new species from clinical specimens from cats. Int. J. Syst. Bacteriol. 39, 373–377.

    Article  Google Scholar 

  • Layer, F., Ghebremedhin, B., König, W., and König, B. 2007. Differentiation of Staphylococcus spp. by terminal-restriction fragment length polymorphism analysis of glyceraldehyde-3-phosphate dehydrogenase-encoding gene. J. Microbiol. Methods 70, 542–549.

    Article  PubMed  CAS  Google Scholar 

  • Lee, Y.D., Moon, B.-Y., Park, J.-H., Chang, H.-I., and Kim, W.J. 2007. Expression of enterotoxin genes in Staphylococcus aureus isolates based on mRNA analysis. J. Microbiol. Biotechnol. 17, 461–467.

    PubMed  CAS  Google Scholar 

  • Lilenbaum, W., Nunes, E.L.C., and Azeredo, M.A.I. 1998. Prevalence and antimicrobial susceptibility of staphylococci isolated from the skin surface of clinically normal cats. Lett. Appl. Microbiol. 27, 224–228.

    Article  PubMed  CAS  Google Scholar 

  • Matthews, K.R., Harmon, R.J., and Langlois, B.E. 1991. Effect of naturally occurring coagulase-negative staphylococci infections on new infections by mastitis pathogens in the bovine. J. Dairy Sci. 74, 1855–1859.

    Article  PubMed  CAS  Google Scholar 

  • Mendoza, M., Meugnier, H., Bes, M., Etienne, J., and Freney, J. 1998. Identification of Staphylococcus species by 16S–23S rDNA intergenic spacer PCR analysis. Int. J. Syst. Bacteriol. 48, 1049–1055.

    Article  PubMed  CAS  Google Scholar 

  • Moodley, A. and Guardabassi, L. 2009. Clonal spread of methicillin-resistant coagulase-negative staphylococci among horses, personnel and environmental sites at equine facilities. Vet. Microbiol. 137, 397–401.

    Article  PubMed  Google Scholar 

  • Nováková, D., Sedlácek, I., Pantůcek, R., Stetina, V., Svec, P., and Petrás, P. 2006. Staphylococcus equorum and Staphylococcus succinus isolated form human clinical specimens. J. Med. Microbiol. 55, 523–528.

    Article  PubMed  Google Scholar 

  • Oliveira, D.C. and de Lencastre, H. 2002. Multiplex PCR strategy for rapid identification of structural types and variants of the mec element in methicillin-resistant Staphylococcus aureus. Antimicrob. Agents Chemother. 46, 2155–2161.

    Article  PubMed  CAS  Google Scholar 

  • Onni, T., Sanna, G., Cubeddu, G.P., Marogna, G., Lollai, S., Leori, G., and Tola, S. 2010. Identification of coagulase-negative staphylococci isolated from ovine milk samples by PCR-RFLP of 16S rRNA and gap genes. Vet. Microbiol. 144, 347–352.

    Article  PubMed  CAS  Google Scholar 

  • Renneberg, J., Rieneck, K., and Gutschik, E. 1995. Evaluation of Staph ID 32 system and Staph-Zym system for identification of coagulase-negative staphylococci. J. Clin. Microbiol. 33, 1150–1153.

    PubMed  CAS  Google Scholar 

  • Sampimon, O.C., Zadoks, R.N., De Vliegher, S., Supré, K., Haesebrouck, F., Barkema, H.W., Sol, J., and Lam, T.J. 2009. Performance of API Staph ID 32 and Staph-Zym for identification of coagulase-negative staphylococci isolated from bovine milk samples. Vet. Microbiol. 136, 300–305.

    Article  PubMed  CAS  Google Scholar 

  • Tsubakishita, S., Kuwahara-Arai, K., Sasaki, T., and Hiramatsu, K. 2010. Origin and molecular evolution of the determinant of methicillin resistance in staphylococci. Antimicrob. Agents Chemother. 54, 4352–4359.

    Article  PubMed  CAS  Google Scholar 

  • Yasuda, R., Kawano, J., Matsuo, E., Masuda, T., Shimizu, A., Anzai, T., and Hashikura, S. 2002. Distribution of mecA-harboring staphylococci in healthy mares. J. Vet. Med. Sci. 64, 821–827.

    Article  PubMed  CAS  Google Scholar 

  • Yasuda, R., Kawano, J., Onda, H., Talagi, M., Shimizu, A., and Anzai, T. 2000. Methicillin-resistant coagulase-negative staphylococci isolated from healthy horses in Japan. Am. J. Vet. Res. 61, 1451–1455.

    Article  PubMed  CAS  Google Scholar 

  • Yugueros, J., Temprano, A., Berzal, B., Sanchez, M., Hernanz, C., Luengo, J.M., and Naharro, G. 2000. Glyceraldehyde-3-phosphate dehydrogenase-encoding gene as a useful taxonomic tool for Staphylococcus spp. J. Clin. Microbiol. 38, 4351–4355.

    PubMed  CAS  Google Scholar 

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Correspondence to Karol Fijałkowski.

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Karakulska, J., Fijałkowski, K., Nawrotek, P. et al. Identification and methicillin resistance of coagulase-negative staphylococci isolated from nasal cavity of healthy horses. J Microbiol. 50, 444–451 (2012). https://doi.org/10.1007/s12275-012-1550-6

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