A Staphylococcus xylosus isolate with a new mecC allotype.

Recently, a novel variant of mecA known as mecC (mecA(LGA251)) was identified in Staphylococcus aureus isolates from both humans and animals. In this study, we identified a Staphylococcus xylosus isolate that harbors a new allotype of the mecC gene, mecC1. Whole-genome sequencing revealed that mecC1 forms part of a class E mec complex (mecI-mecR1-mecC1-blaZ) located at the orfX locus as part of a likely staphylococcal cassette chromosome mec element (SCCmec) remnant, which also contains a number of other genes present on the type XI SCCmec.

Characterization of methicillin-resistant Staphylococcus aureus CC398 obtained from humans and animals on dairy farms.

In this study MRSA isolates from dairy farms were investigated for their genetic relationships and antimicrobial susceptibility. In total, 125 MRSA isolates from 26 dairy farms were studied, including isolates from milk samples (n=46), dairy cattle (n=24), calves (n=6), dust samples from pig (n=16) and veal calf sheds (n=1), dogs (n=2), a horse, a sheep and humans (n=28). CC398-specific PCRs, spa typing, SCCmec typing and ApaI macrorestriction analysis were conducted. Susceptibility testing was performed by broth microdilution. All 125 isolates belonged to CC398. Eight spa types (t011, t108, t034, t567, t1184, t1451, t2287 and t3934) were detected. SCCmec elements of types IV (n=48) and V (n=67) were identified with 10 isolates being non-typeable. Six main macrorestriction patterns - with up to 23 sub-patterns - and twelve resistance patterns were identified. Sixty-eight isolates showed a multiresistance phenotype. Farm-by-farm analysis revealed different scenarios: in some farms, the MRSA CC398 isolates from dairy cattle, humans, pig sheds and/or sheep were indistinguishable suggesting an interspecies exchange of the same MRSA CC398 subtype. In other farms, several MRSA CC398 subtypes were detected in different host species/sources with occasionally even more than one MRSA CC398 subtype from the same host species/source. These latter results may suggest that either different MRSA subtypes associated with humans or animals have been imported into the respective farm or that one MRSA CC398 strain has undergone diversification, reflected by more or less expanded changes in PFGE patterns, spa type or resistance pattern, during colonization of different hosts on the same farm.

Bovine-associated MRSA ST398 in the Netherlands.

During routinely screening (50.000 milk samples on an annual basis) 14 MRSA ST398 strains were identified in the period of January 2008 to September 2008 in 14 different dairy herds located in the provinces Overijssel and Gelderland, The Netherlands. Molecular analysis was performed by Cfr9I PFGE, ST398-specific diagnostic PCR, spa typing, SCCmec typing and Panton-Valentine Leukocidin (PVL) gene PCR. The molecular analyses of 14 MRSA (one MRSA strain per herd) strains revealed that all strains belong to ST398 with 3 closely related spa types (t011, t108 and t889, all commonly found in pigs) and carry 2 different SCCmec types, IVa and V. All MRSA strains were resistant to two or more classes of antibiotics and also PVL negative. The majority of farms (n = 9, 64%) harboured combined livestock with both cows and pigs present. Our study contributes to the growing evidence that MRSA ST398 is transmitted among various animal species and can be considered as an etiological agent of mastitis in dairy cows.

Staphylococcus devriesei sp. nov., isolated from teat apices and milk of dairy cows.

Ten non-motile, Gram-stain-positive, coagulase-negative staphylococci were isolated from bovine milk and teat apices. All isolates were catalase-positive, with anteiso-C(15 : 0), iso-C(15 : 0), anteiso-C(17 : 0), iso-C(17 : 0) and C(18 : 0) as predominant fatty acids and diphosphatidylglycerol and phosphatidylglycerol as major polar lipids. The results of sequence analysis of the 16S rRNA gene and four housekeeping genes (rpoB, hsp60, tuf and dnaJ) in combination with tRNA-intergenic spacer length analysis showed that the isolates form a separate branch within the genus Staphylococcus. Based on 16S rRNA gene sequencing, the phylogenetically most closely related species are Staphylococcus haemolyticus, S. hominis and S. lugdunensis, with >98.7 % sequence similarity. The DNA G+C content varies from 33.3 to 33.7 mol%, and DNA-DNA hybridization with the nearest neighbours, based on 16S rRNA gene sequences, confirmed that the isolates represent a novel Staphylococcus species. All isolates induced a small zone of complete haemolysis on Columbia agar with 5 % sheep blood and exhibited a homogeneous biochemical fingerprint that is discriminative from the phylogenetically most closely related species. Based on these results, it is proposed to classify the ten isolates as Staphylococcus devriesei sp. nov., with strain KS-SP 60(T) (=LMG 25332(T) =CCUG 58238(T)) as the type strain.