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This observational study aimed to determine MRSA prevalence using strain‐specific real‐time PCR at the pig level, stratified by age groupings, within a pig enterprise. A total of 658 samples were collected from individual pigs (n = 618) and the piggery environment (n = 40), distributed amongst five different pig age groups. Presumptive MRSA isolates were confirmed by the presence of mecA, and MALDI‐TOF was performed for species verification. All isolates were tested against 18 different antimicrobials. MRSA was isolated from 75.2% (95% CI 71.8–78.6) of samples collected from pigs, and 71% of the MRSA isolates from this source were identified as community‐associated (CA)‐MRSA ST93, while the remainder were livestock‐associated (LA)‐MRSA ST398. Amongst environmental isolates, 80% (CI 64.3–95.7) were ST93 and the remainder ST398. All MRSA isolates from pigs and the environment were susceptible to ciprofloxacin, gentamicin, linezolid, mupirocin, rifampicin, sulfamethoxazole–trimethoprim, teicoplanin and vancomycin. Phenotypic rates of resistance were penicillin (100%), clindamycin (97.6%), erythromycin (96.3%), ceftiofur (93.7%), chloramphenicol (81.2%), tetracycline (63.1%) and amoxicillin–clavulanate (63.9%). A low prevalence of resistance (9.2%) was observed against neomycin and quinupristin–dalfopristin. The probability of MRSA carriage in dry sows (42.2%) was found to be significantly lower (p <.001) when compared to other age groups: farrowing sows (76.8%, RR1.82), weaners (97.8%, RR 2.32), growers (94.2%, RR 2.23) and finishers (98.3%, RR 2.33). Amongst different production age groups, a significant difference was also found in antimicrobial resistance for amoxicillin–clavulanate, neomycin, chloramphenicol and tetracycline. Using the RT‐PCR assay adopted in this study, filtering of highly prevalent ST93 and non‐ST93 isolates was performed at high throughput and low cost. In conclusion, this study found that weaner pigs presented a higher risk for CA‐MRSA and antimicrobial resistance compared to other age groups. These findings have major implications for how investigations of MRSA outbreaks should be approached under the One‐Health context. [ABSTRACT FROM AUTHOR]

Background: The Central Australian Indigenous population has a high incidence of Staphylococcus aureus bacteremia (SAB) but little is known about the local molecular epidemiology.Methods: Prospective observational study of bacteremic and nasal colonizing S.aureus isolates between June 2006 to June 2010. All isolates underwent single nucleotide polymorphism (SNP) genotyping and testing for the presence of the Panton-Valentine Leucocidin (pvl) gene.Results: Invasive isolates (n = 97) were predominantly ST93 (26.6 %) and pvl positive (54.3 %), which was associated with skin and soft tissue infections (OR 4.35, 95 % CI 1.16, 16.31). Non-multiresistant MRSA accounted for 31.9 % of bacteremic samples and showed a trend to being healthcare associated (OR 2.16, 95 % CI 0.86, 5.40). Non-invasive isolates (n = 54) were rarely ST93 (1.9 %) or pvl positive (7.4 %).Conclusions: In Central Australia, ST93 was the dominant S.aureus clone, and was frequently pvl positive and associated with an aggressive clinical phenotype. Whether non-nasal carriage is more important with invasive clones or whether colonization occurs only transiently remains to be elucidated. [ABSTRACT FROM AUTHOR]

High-resolution melting analysis is an inherently robust, easy and inexpensive approach to the examination of genomic regions containing single-nucleotide polymorphisms and hypervariable loci. Staphylococcus aureus sequence type (ST) 93 is a singleton, Panton–Valentine leukocidin-positive clone unique to Australia. A high-resolution melting-based method for the identification of ST93 was developed, and a similar approach was used to reveal diversity within the spa locus of this lineage. Statistical and graphical methods that account for instrumental and operator-dependent variation in high-resolution melting curves were developed, to allow greater confidence and reproducibility in deciding whether another curve is truly different from the baseline curve of an amplicon with known sequence. The data support a very early acquisition, or multiple independent acquisitions, of SCC mec by ST93 methicillin-susceptible S. aureus (MSSA), and the coexistence of MSSA and methicillin-resistant S. aureus versions of the same lineage within northern Australia. [ABSTRACT FROM AUTHOR]