Antimicrobial resistance of coagulase-negative staphylococci from bovine subclinical mastitis with particular reference to macrolide–lincosamide resistance phenotypes and genotypes

Abstract

The aim of this study was to analyse coagulase-negative staphylococci (CoNS) for their resistance to antimicrobial agents approved for the control of pathogens involved in bovine mastitis, with particular reference to macrolide and/or lincosamide (ML) resistance and the resistance genes involved. A total of 298 CoNS collected between 2003 and 2005 in Germany from cases of subclinical mastitis in dairy cows were identified to the species level and investigated for their MICs by broth microdilution. ML-resistant isolates were subjected to plasmid profiling and electrotransformation experiments. The ML resistance genes were detected using PCR and hybridization. Selected PCR products were cloned and sequenced. The CoNS isolates used in this study showed a low level of resistance to all antimicrobial agents tested (0-7.4%) except ampicillin (18.1%). In the erythromycin-resistant and/or pirlimycin-resistant isolates, the ML resistance genes erm(B), erm(C), msr(A), mph(C) and lnu(A) were present, either alone or in different combinations. Isolates carrying erm methylase genes or the exporter gene msr(A) showed higher MICs than those harbouring only the genes mph(C) or lnu(A) coding for inactivating enzymes. Most of the ML resistance genes were found on plasmids. This is the first report of pirlimycin MICs for CoNS collected from cases of bovine subclinical mastitis in Germany. After 3-5 years of veterinary therapeutic use, pirlimycin resistance was rarely detected among CoNS. The finding that five different resistance genes--present in various combinations--were responsible for ML resistance underlines the heterogeneous character of this resistance trait.