Role of staphylococci in mastitis in sheep.

Staphylococci have been isolated from various sites of the body of healthy sheep, as well as from many infections of those animals, the main one being mastitis. The objective of this review is to appraise the importance and significance of staphylococci in causing mastitis in ewes. The review includes a brief classification and taxonomy of staphylococci and describes the procedures for their isolation and identification, as well as their virulence determinants and the mechanisms of resistance to antibacterial agents. Various staphylococcal species have been implicated in staphylococcal mastitis and the characteristics of isolates are discussed with regards to potential virulence factors. Staphylococcal mastitis is explicitly described, with reference to sources of infection, the course of the disease and the relevant control measures. Finally, the potential significance of staphylococci present in ewes' milk for public health is discussed briefly.

Antimicrobial Agent Susceptibility and Typing of Staphylococcal Isolates from Subclinical Mastitis in Ewes.

Objective was to study susceptibility to antimicrobial agents of 142 staphylococcal isolates from subclinical mastitis in ewes. In total, 41.5% of these were resistant and 5.6% multidrug resistant. More coagulase-negative staphylococci (47.0%) were resistant than Staphylococcus aureus (18.5%) isolates. Resistance was greater to penicillin (22.5%), tetracycline, or ampicillin (18.3%). More biofilm-forming (20.6%) isolates were resistant to tetracycline than nonbiofilm-forming (0.0%) ones. Presence of tetK was associated with presence of icaA in the same strains. Further, 76.6% of resistant isolates versus 57.7% of susceptible ones were recovered immediately postpartum and 23.4% of resistant isolates versus 9.9% of susceptible ones were recovered in farms that practiced routine administration of antimicrobial agents at the end of a lactation period. Most S. aureus (59.3%) were classified in ST133 and most Staphylococcus epidermidis were classified in ST100, ST142, or ST152 (19.0% each). There was no association of sequence types with resistance. Whole genome sequencing showed that, in a Staphylococcus lentus strain, the ermB gene was part of transposon Tn917 integrated into the chromosome; also, a small plasmid was observed in an ermC-carrying Staphylococcus hominis strain and, finally, in an S. aureus and an S. epidermidis strains, small tetK-carrying plasmids (pSau-2716Lar, pSau-3893Lar) of 4.439 kb were found.

MLSB-Resistant Staphylococcus aureus in Central Greece: Rate of Resistance and Molecular Characterization.

The aim of the present study was to determine the rate and mechanisms of resistance to macrolides, lincosamides, and streptogramin B (MLSB) antibiotics of Staphylococcus aureus collected in Central Greece. Of the 2,893 S. aureus collected during 2012-2017, 1,161 isolates (40.2%) exhibited resistance to at least one of the MLSB agents. The rate of erythromycin resistance was statistically significantly higher in methicillin-resistant S. aureus (MRSA) (58.6%) than in methicillin-sensitive S. aureus (MSSA) isolates (20.7%) (p = 0.002). Two hundred seventy-five representative MLSB-resistant S. aureus, including 81 MSSA and 194 MRSA isolates, were further studied. Thirty-eight MSSA isolates carried ermC, 26 MSSA were positive for ermA, whereas 17 isolates carried msrA gene. Among MRSA, the ermA gene was identified in the majority of the isolates (n = 153). Thirty-seven MRSA isolates carried ermC; three isolates carried msrA, whereas the remaining MRSA was positive for two genes (ermA and ermC). Phylogenetic analysis showed that ST225, which belongs to CC5, was the most prevalent, accounting for 137 MRSA isolates. Higher genetic diversity was found in the group of MSSA isolates, which comprised of 13 sequence types. Whole-genome sequencing data showed that all ermA-positive S. aureus, with the exception of one ST398 isolate, harbored the ermA-carrying Tn554 transposon integrated into their chromosomes. Furthermore, Illumina sequencing followed by polymerase chain reaction screening identified that ermC, which was identified in a polyclonal population of MSSA and MRSA isolates, was carried by small plasmids, like pNE131. These findings highlighted the important role of high-risk clones and of mobile elements carrying resistance genes in the successful dissemination of MLSB-resistant staphylococci.

First Description in Greece of mphC-Positive Staphylococci Causing Subclinical Mastitis in Ewes.

The aim of the present study was to describe the first mphC-positive staphylococci, including two Staphylococcus lentus (Sle-087lar and Sle-091lar) and one Staphylococcus xylosus (Sxy-228lar), isolated from samples of animal origin, in Greece. Isolates Sle-087lar and Sxy-228lar were resistant to erythromycin, whereas Sle-091lar was resistant to erythromycin and lincomycin. All three isolates were susceptible to the remaining antibiotics. PCR screening showed that isolate Sle-091lar carried also ermB. For Sxy-228lar, whole-genome sequencing (WGS) and de novo assembly obtained an mphC-positive contig of 57.3-kb exhibiting high similarity with the genome of mphC-negative S. xylosus S170. However, mphC of Sxy-228lar was 91% similar to that found in plasmid pJW2311 from S. xylosus JW2311. Additionally, WGS data showed that Sle-087lar and Sle-091lar harbored mphC-carrying sequences being highly similar to the recently announced genome of the mphC-carrying S. lentus isolate 050AP from Tanzania. However, differences were observed in the mphC environment, suggesting the independent acquisition of the gene by each isolate. Sle-091lar also harbored transposon Tn917, which carries ermB resistance gene, integrated into S. lentus chromosome. These findings indicated that acquisition of resistance genes can lead to the emergence of multiresistant staphylococci, causing animal infections with economic burden.