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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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.
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.
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.
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.
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.
CLSI. 2008. Performance standards for antimicrobial susceptibility testing; 18th informational supplement. CLSI document M100-S18. Clinical and Laboratory Standards Institute, Wayne, PA, USA.
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.
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.
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.
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.
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.
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.
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.
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.
Harmsen, D. and Karch, H. 2004. 16S rDNA for diagnosing pathogens: a living tree. ASM News 70, 19–24.
Harmsen, D., Rothgänger, J., Frosch, M., and Albert, J. 2002. RIDOM: ribosomal differentiation of medical micro-organisms database. Nucleic Acids Res. 30, 416–417.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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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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DOI: https://doi.org/10.1007/s12275-012-1550-6