Phenotypic and genotypic antimicrobial resistance of staphylococci from bovine milk

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Abstract

The aim of this study was to examine phenotypic and genotypic antimicrobial resistance of staphylococci from milk samples from cows with subclinical and clinical mastitis and from cows without mastitis symptoms to methicillin, tetracyclines, macrolides and lincosamides (ML). Of 207 strains, including 34 S. aureus and 173 coagulase-negative staphylococci (CNS), 11 (6.4%) CNS strains were phenotypically resistant to methicillin. The mecA gene was detected by PCR only in two S. xylosus strains and one strain of S. epidermidis and S. simulans. No methicillin-resistant S. aureus strains were observed. In methicillin-resistant strains with mecA, gene resistance to other investigated antibiotics was not observed. Phenotypic resistance to tetracycline was detected in 11.0% of CNS strains and 47.4% of them carried the tetK gene. Of 173 CNS strains studied, 27 (15.6%) were resistant to at least one ML antibiotic. The resistance gene ermC was detected in 55.5% of the 27 ML-resistant strains. The ermA and ermB genes were detected in 14.8% and 11.1% of ML-resistant CNS strains, respectively. Antimicrobial resistance to methicillin, tetracyclines and macrolides was detected more frequently in staphylococcal strains from clinical mastitis compared to animals with subclinical symptoms and without mastitis, while the resistance to lincosamides showed a similar frequency in all groups of cows. In conclusion, CNS species from bovine milk differ in phenotypic and genotypic antimicrobial resistance profiles, and the use of PCR technique alone for the detection of methicillin, macrolide, lincosamide and tetyracycline resistance in CNS from cattle is not reliable.

Bannerman TL (2003) Staphylococcus, micrococcus, and other catalase-positive cocci that grow aerobically. In: Murray PR, Baron EJ, Jorgensen JH, Pfaller MA, Yolken RH (eds) Manual of clinical microbiology. American Society Microbiology, Washington, pp 384-404.

Broekema NM, Van TT, Monson TA, Marshall SA, Warshauer DM (2009) Comparison of cefoxitin and oxacillin disc diffusion methods for detection of mecA-mediated resistance in Staphylococcus aureus in a large-scale study. J Clin Microbiol 47: 217-219.

Capurro A, Concha C, Nilsson L, O¨ stensson K (1999) Identification of coagulase-positive staphylococci isolated from bovine milk. Acta Vet Scand 40: 315-321.

Casadevall A, Pirofski LA (2000) Host-pathogen interactions: basic concepts of microbial commensalism, colonization, infection and disease. Infect Immun 68: 6511-6518.

Chopra I, Roberts M (2001) Tetracycline antibiotics: mode of action, applications, molecular biology and epidemiology of bacterial resistance. Microbiol Mol Biol Rev 65: 232-260.

Clinical and Laboratory Standards Institute (CLSI) (2011) Performance Standards for Antimicrobial Susceptibility Testing; Twenty-First Informational Supplement. CLSI, Wayne, M100-S21, 31 No. 1.

Gillespie BE, Headrick SI, Boonyayatra S, Oliver SP (2009) Prevalence and persistence of coagulase-negative Staphylococcus species in three dairy research herds. Vet Microbiol 134: 65-72.

Khan AA, Nawaz MS, Khan SA, Steele R (2002) Detection and characterization of erythromycin-resistant methylase genes in Gram-positive bacteria isolated from poultry litter. Appl Microbiol Biotechnol 59: 377-381.

Leclercq R, Bauduret F, Soussy CJ (1989) Selection of constitutive mutants of gram-positive cocci inducible resistant to macrolides, lincosamides and streptogramins (MLS): comparison of the selective effects of the MLS. Pathol Biol (Paris) 37: 568-572.

Lina G, Quaglia A, Reverdy ME, Leclercq R, Vandenesch F, Etienne J (1999) Distribution of genes encoding resistance to macrolides, lincosamides, and streptogramins among staphylococci. Antimicrob Agents Chemother 43: 1062-1066.

Lüthje P, Schwarz S (2006) Antimicrobial resistance of coagulase- negative staphylococci from bovine subclinical mastitis with particular reference to macrolide-lincosamide resistance phenotypes and genotypes. J Antimicrob Chemother 57: 966-969.

Malinowski E, Kłossowska A, Kaczmarowski M, Lassa H, Kuźma K (2002) Antimicrobial susceptibility of staphylococci isolated from affected with mastitis cows. Bull Vet Inst Pulawy 46: 289-294.

Malinowski E, Lassa H, Kłossowska A, Smulski S, Markiewicz H, Kaczmarowski M (2006a) Etiological agents of dairy cows’ mastitis in western part of Poland. Pol J Vet Sci 9: 191-194.

Malinowski E, Lassa H, Kłossowska A, Markiewicz H, Kaczmarowski M, Smulski S (2006b) Relationship between mastitis agents and somatic cell count in foremilk samples. Bull Vet Inst Pulawy 50: 349-352.

Martin B, Garriga M, Hugas M, Bover-Cid S. Veciana-Nogue´s MT, Aymerich T (2006) Molecular, technological and safety characterization of Gram-positive catalase-positive cocci from slightly fermented sausages. Int J Food Microbiol 107: 148-158.

Merlino J, Watson J, Rose B, Beard-Pegler M, Gottlieb T, Bradbury R, Harbour C (2002) Detection and expression of methicillin/oxacillin resistance in multidrug-resistant and non- multidrug-resistant Staphylococcus aureus in Central Sydney, Australia. J Antimicrob Chemother 49: 793-801.

Rajala-Schultz PJ, Torres AH, Degraves FJ, Gebreyes WA, Patchanee P (2009) Antimicrobial resistance and genotypic characterization of coagulase-negative staphylococci over the dry period. Vet Microbiol 134: 55-64.

Resch M, Nagel V, Hertel C (2008) Antibiotic resistance of coagulase-negative staphylococci associated with food and used in starter cultures. Int J Food Microbiol 127: 99-104.

Sampimon OC, Barkema HW, Berends IM, Sol J, Lam TJ (2009a) Prevalence and herd-level risk factors for intramammary infection with coagulase-negative staphylococci in Dutch dairy herds. Vet Microbiol 134: 37-44.

Sampimon OC, Zadoks RN, De Vliegher S, Supre´ K, Haesebrouck F, Barkema HW, Sol J, Lam TJ (2009b) 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.

Sawant AA, Gillespie BE, Oliver SP (2009) Antimicrobial susceptibility of coagulase-negative Staphylococcus species isolated from bovine milk. Vet Microbiol 134: 73-81.

Sawant AA, Sordillo LM, Jayarao BM (2005) A survey on antibiotic usage in dairy herds in Pennsylvania. J Dairy Sci 88: 2991-2999.

Simeoni D, Rizzotti L, Cocconcelli P, Gazzola S, Dellaglio F, Torriani S (2008) Antibiotic resistance genes and identification of staphylococci collected from the production chain of swine meat commodities. Food Microbiol 25: 196-201.

Strommenger B, Kettlitz C, Werner G, Witte W (2003) Multiplex PCR assay for simultaneous detection of nine clinically relevant antibiotic resistance genes in Staphylococcus aureus. J Clin Microbiol 41: 4089-4094.

Suzuki E, Hiramatsu K, Yokota T (1992) Survey of methicillin- resistant clinical strains of coagulase-negative staphylococci for mecA gene distribution. Antimicrob Agents Chemother 36: 429-434.

Taponen S, Pyörälä S (2009) Coagulase-negative staphylococci as cause of bovine mastitis-not so different from Staphylococcus aureus? Vet Microbiol 134: 29-36.

Taponen S, Simojoki H, Haveri M, Larsen HD, Pyo¨ra¨la¨ S (2006) Clinical characteristics and persistence of bovine mastitis caused by different species of coagulase-negative staphylococci identified with API or AFLP. Vet Microbiol 115: 199-207.

Tenover FC, Jones RN, Swenson JM, Zimmer B, McAllister S, Jorgensen JH (1999) Methods for improved detection of oxacillin resistance in coagulase-negative staphylococci: results of a multicenter study. J Clin Microbiol 37: 4051-4058.

Trieu-Cuot P, Poyart-Salmeron C, Carlier C, Courvalin P (1990) Nucleotide sequence of the erythromycin resistance gene of the conjugative transposon Tn1545. Nucleic Acids Res 18: 3660.

Trzcinski K, Cooper BS, Hryniewicz W, Dowson CG (2000) Expression of resistance to tetracyclines in strains of methicillin- resistant Staphylococcus aureus. J Antimicrob Chemother 45: 763-770.

Walther C, Perreten V (2007) Methicillin-resistant Staphylococcus epidermidis in organic milk production. J Dairy Sci 90: 5351.

Werckenthin C, Schwarz S, Westh H (1999) Structural alterations in the translational attenuator of constitutively expressed ermC genes. Antimicrob Agents Chemother 43: 1681-1685.

York MK, Gibbs L, Chehab F, Brooks GF (1996) Comparison of PCR detection of mecA with standard susceptibility testing methods to determine methicillin resistance in coagulase-negative staphylococci. J Clin Microbiol 34: 249-253.

Polish Journal of Veterinary Sciences

The Journal of Committee of Veterinary Sciences of Polish Academy of Sciences and University of Warmia and Mazury in Olsztyn

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