Phenotypical and Genotypical Antimicrobial Resistance of Coagulase-negative staphylococci Isolated from Cow Mastitis

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The objectives of this study were to determine the prevalence and antimicrobial resistance of coagulase-negative staphylococci (CNS) isolated from dairy cows with subclinical mastitis. Antimicrobial resistance in staphylococci were evaluated by breakpoint values specific to the species (EU-CAST). The presence of resistance-encoding genes was detected by multiplex PCR. A total of 191 CNS isolates were obtained. The CNS isolates were typically resistant to penicillin (67.4%), tetracyc-line (18.9%), and erythromycin (13.7%). CNS isolates (78.0%) were resistant to at least one antimicrobial compound, and 22.0% were multiresistant. The multiresistant isolates were predominantly Staphylococcus chromogenes (28.6%), Staphylococcus warneri (19%) and Staphylococcus haemolyticus (14.3%). According to MIC pattern data, multiresistant isolates showed the highest resistance (p<0.05) rates to penicillin (85.7%), tetracycline (66.7%), and erythromycin (48.2%), but all of them were sensitive to daptomycin, oxacillin, qiunupristin/dalfopristin, and vancomycin. S. chromogenes (9.5%), S. haemolyticus (4.8%), and S. capitis ss capitis (2.4%) strains were resistant to methicillin; their resistance to oxacillin and penicillin was more than 8 mg/l. A high rate of resistance to penicillin was linked to a blaZ gene found in 66.6% of the isolated multiresistant CNS strains. Resistance to tetracycline via the tetK (38.1%) gene and penicillin via the mecA (23.8%) gene were detected less frequently. Gene msrAB was responsible for macrolides and lincosamides resistance and detected in 28.6% of the CNS isolates. Antimicrobial resistance genes were identified more frequently in S. epidermidis, S. chromogenes, and S. warneri.

Aarestrup FM (ed), (2006) Antimicrobial Resistance. In: Bacteria of Animal Origin. ASM Press, Washington, DC, p 187-206.

Aerestrup FM, Agersø Y, Ahrens P, Østergaard Jzrgensen JC, Madsen M, Jensen LB (2000) Antimicrobial susceptibility and presence of resistance genes in staphylococi from poultry. Vet Microbiol 74: 353-364.

Anonymous (2008) Multiplex PCR for the detection of the mecA gene and the identification of Staphylococcus aureus. In: National Food Institute (DTU) Protocol. National Food Institute, Technical University of Denmark, Copenhagen.

Clinical and Laboratory Standarts Institute (2010) Performance standards for antimicrobial disk susceptibility tests. Approved standard. M2-A9 Villanova, Pa: Clinical and Laboratory Standards Institute.

Couto I, Pereira S, Miragaia M, Sanches I S, de Lencastre H (2001) Identification of clinical staphylococcal isolates from humans by internal transcribed spacer PCR. J. Clin. Microbiol 39: 3099-3103.

Cuny C, Layer F, Strommenger B, Witte W (2011) Rare occurrence of methicillin-resistant Staphylococcus aureus CC130 with a novel mecA homologue in humans in Germany. PloS One 6: e24360.

Dubois D, Grare M, Prere MF, Segonds C, Marty N, Oswald E (2012) Performances of the Vitek MS matrix-assisted laser desorption ionization-time of flight mass spec-trometry system for rapid identification of bacteria in routine clinical microbiology. J Clin Microbiol 50: 2568-2576.

Fessler AT, Billerbeck C, Kadlec K, Schwarz S (2010) Identification and characterization of methicillin-resistant coagulase-negative staphylococci from bovine mastitis. J. Antimicrob. Chemother 65: 1576-1582.

Frey Y, Rodriguez JP, Thomann A, Schwendener S, Perreten V (2013) Switzerland Genetic characterization of antimicrobial resistance in coagulase-negative staphylococci from bovine mastitis milk. J. Dairy Sci 96: 2247-2257.

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.

Jensen AG, Wachmann CH, Espersen F, Scheibel J, Skinhoj P, Frimdt-Moller N (2002) Treatment and outcome of Staphylococcus aureus bacteremia: a prospective study of 278 cases. Arch Intern Med 162: 25 32.

Kadlec K, Schwarz S (2010) Identification of a plasmid-borne resistance gene cluster comprising the resistance genes erm(T), dfrK, and tet(L) in a porcine methicillin-resistant Staphylococcus aureus ST398 strain. Antimicrob Agents Chemother 54: 915-918.

Klimiene I, Ružauskas M, Špakauskas V, Matusevičius A, Mockeliūnas R, Pereckienė A, Butrimaitė-Ambrozevičienl Č, Virgailis M (2011) Antimicrobial resistance patterns to beta-lactams of gram-positive cocci isolated from bovine mastitis in Lithuania. Pol J Vet Sci 14: 467-472.

Kot B, Piechota M, Wolska KM, Frankowska A, Zdunek E, Binek T, K opotowska E, Antosiewicz M (2012) Phenotypic and genotypic antimicrobial resistance of staphylococci from bovine milk. Pol J Vet Sci 15: 677-83.

Malinowski E, Kłossowska A, Zastempowska E (2011) Virulence factors in coagulase-negative staphylococci isolated from cows with subclinical mastitis. Bull Vet Inst Pulawy 55: 681-684.

Perreten V, Vorlet-Fawer L, Slickers P, Ehricht R, Kuhnert P, Frey J (2005) Microarray-based detection of 90 antibiotic resistance genes of gram-positive bacteria. J Clin Microbiol 43: 2291-2302.

Piessens V, Van Coillie E, Verbist B, Supré K, Braem G, Van Nuffel A, De Vuyst L, Heyndrickx M, De Vliegher S (2011) Distribution of coagulase-negative Staphylococcus species from milk and environment of dairy cows differs between herds. J Dairy Sci 94: 2933-2944.

Pyörälä S, Taponen S (2009) Coagulase-negative staphylococci-Emerging mastitis pathogens. Vet Microbiol 134: 3-8.

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

Schnellmann C, Gerber V, Rossanno A, Jaquier V, Panchaud Y, Doherr M, Thomann A, Straub R, Perreten V (2006) Presence of new mecA and mph(c) variants conferring antibiotic resistance in Staphylococcus spp. Isolated from the skin of horses before and after clinic admission. J Clin Microbiol 44: 4444-4454.

Skold O (2001) Resistance to trimethoprim and sulfonamides. Vet Res 32: 261-273.

Soares LC, Pereira IA, Pribul BR, Oliva MS, Coelho S, Souza MMS (2012) Antimicrobial resistance and detection of mecA and blaZ genes in coagulase-negative Staphylococcus isolated from bovine mastitis. Pesq Vet Bras 32: 692-696.

Supré K, Haesebrouck F, Zadoks RN, Vaneechoutte M, Piepers S, De Vliegher S (2011) Some coagulase-negative Staphylococcus species affect udder health more than others. J Dairy Sci 94: 2329-2340.

Sztachanska M, Baranski W, Janowski T, Pogorzelska JZdunczyk S (2016) Prevalence and etiological agents of subclinical mastitis at the end of lactation in nine dairy herds in North-East Poland. Pol J Vet Sci 19 (1): 119-124.

Thorberg BM, Danielsson-Tham ML, Emanuelson U, Persson Waller K (2009) Bovine subclinical mastitis caused by different types of coagulase-negative staphylococci. J Dairy Sci 92: 4962-4970.

Thumu SC, Halami PM (2012) Presence of erythromycin and tetracycline resistance genes in lactic acid bacteria from fermented foods of Indian origin. Antonie Van Leeuwenhoek 102: 541-551.

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

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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