Sociodemographic and molecular epidemiological associations of community-associated MRSA (CA-MRSA) and healthcare-associated MRSA (HCA-MRSA) in New Zealand, 2005–2011.

<p>Note: Data are number (%) of patients unless stated otherwise.</p>a<p>Mann-Whitney <i>U</i> test.</p><p>Abbreviations: OR, odds ratio; CI, confidence interval; IQR, interquartile range; ST, sequence type, NZDep, New Zealand Deprivation Index score; MRSA, methicillin-resistant <i>Staphylococcus aureus</i>.</p

Methicillin-resistant <i>Staphylococcus aureus</i> (MRSA) period-prevalence rates for clinical isolates in New Zealand, 2005–2011, stratified by (A) age, and (B) ethnicity.

<p>Methicillin-resistant <i>Staphylococcus aureus</i> (MRSA) period-prevalence rates for clinical isolates in New Zealand, 2005–2011, stratified by (A) age, and (B) ethnicity.</p

Relative proportions of methicillin-resistant <i>Staphylococcus aureus</i> (MRSA) clones circulating in New Zealand, 2005–2011.

<p>Relative proportions of methicillin-resistant <i>Staphylococcus aureus</i> (MRSA) clones circulating in New Zealand, 2005–2011.</p

Period-prevalence rates of community-associated methicillin-resistant <i>Staphylococcus aureus</i> (CA-MRSA), healthcare-associated community-onset MRSA (HCA-CO MRSA) and healthcare-associated hospital-onset (HCA-HO MRSA) in New Zealand, 2005–2011.

<p>Period-prevalence rates of community-associated methicillin-resistant <i>Staphylococcus aureus</i> (CA-MRSA), healthcare-associated community-onset MRSA (HCA-CO MRSA) and healthcare-associated hospital-onset (HCA-HO MRSA) in New Zealand, 2005–2011.</p

Relative LukF-PV expression in ST93 isolates.

<p>All isolates were tested for LukF-PV expression using western blot and LukF-PV specific antibody. Results are expressed as optical density of test strain relative to a 50 µg control of rLukF-PV that was run on every gel. All experiments were performed with multiple replicates and mean and range is shown. Positive control, rLUKF-PV; negative control, RN4220.</p

Minimum spanning tree (MST) of the six ST93 <i>dru</i> types.

<p>Cluster analysis was performed using the Polymorphic VNTR plug-in tool of the BioNumerics program. <i>dru</i> types separated by an MST distance of ≤1 (i.e., if they were ≥98% similar) were considered closely related and assigned to the same cluster. Pulsed-field gel electrophoresis (PFGE) pulsotypes and <i>spa</i> types are recorded for each <i>dru</i> type.</p

Minimum spanning tree (MST) of the seven ST93 <i>spa</i> types.

<p>Cluster analysis was performed using the spa typing plug-in tool of the BioNumerics program. <i>spa</i> types separated by an MST distance of ≤1 (i.e., if they were ≥98% similar) were considered closely related and assigned to the same cluster. MSSA and MRSA <i>spa</i> types are designated in red and green print respectively. Pulsed-field gel electrophoresis (PFGE) pulsotypes and <i>dru</i> types (dt) are recorded for each <i>spa</i> type.</p

Characterisation of ST93 isolates.

<p>Regions: ACT, Australian Capital Territory; NSW, New South Wales; NT, Northern Territory, Qld, Queensland; SA, South Australia; Vic, Victoria; WA, Western Australia.</p><p>Antibiogram: Ox, oxacillin; Em, erythromycin; Te, tetracycline.</p><p>Resistance Genotype: <i>mecA</i>, alternate penicillin binding protein 2 gene; <i>blaZ</i>, beta lactamase gene; <i>blaI</i>, beta-lactamase repressor (inhibitor) gene; <i>blaR</i> beta-lactamase regulatory protein gene, <i>ermC</i>, erythromycin/clindamycin resistance gene; <i>msr(A)</i>, macrolide resistance gene; <i>tetK</i>, tetracycline resistance gene; <i>dfrA</i>, trimethoprim resistance gene; qacC quaternary ammonium compound resistance gene protein C.</p><p>PFGE, pulsed field gel electrophoresis; MLST, multilocus sequence type; SCCmec, staphylococcal cassette chromosome mec; <i>spa, Staphylococcus aureus</i> protein gene A; <i>dru</i>, the direct-repeat unit (<i>dru</i>) variable-number tandem repeat region adjacent to IS431 in SCC<i>mec.</i></p><p><i>lukF</i>/<i>lukS</i> PV, Panton Valentine leucocidin F and S component genes.</p

Dendrogram of the 58 pulsed-field gel electrophoresis patterns (PFGE) of ST93 (13 MSSA and 45 MRSA).

<p>Sequenced JKD6159 strain was used as the ST93 control. <i>S. aureus</i> strain NCTC 8325 was used as the reference strain.</p

NCL812 compounds exert their antibacterial action on the cell membrane of <i>S</i>. <i>pneumoniae</i> and <i>S</i>. <i>aureus</i>.

<p>(<b>A and B</b>), <i>S</i>. <i>pneumoniae</i> strain D39 exposed to 16 μg/ml NCL812 for 6 h exhibited significantly thicker cell membranes compared to untreated samples (<b>A</b>) (<i>p</i> < 0.0001; two-tailed unpaired <i>t</i>-test) and displayed significantly wider periplasmic space compared to untreated samples (<b>B</b>) (<i>p</i> < 0.001; two-tailed unpaired <i>t</i>-test. Data presented are an example from 12 different bacterial cells, each with at least 10 measurements per bacteria for both treated and untreated samples. (<b>C and D</b>) NCL812 affects macromolecular synthesis (<b>c</b>) and ATP release (<b>D</b>) in <i>S</i>. <i>aureus</i>. (<b>E and F</b>), NCL Compounds dissipate the membrane potential of <i>S</i>. <i>pneumoniae</i> and <i>S</i>. <i>aureus</i>. Membrane potential measurements of <i>S</i>. <i>pneumoniae</i> D39 (<b>E</b>) and <i>S</i>. <i>aureus</i> ATCC49775 (<b>F</b>). Bacterial suspensions were exposed to 16 μg/ml NCL812, NCL195, NCL219, or ampicillin (control) for 5 min after which DiOC₂(3) was added and the fluorescence monitored until it plateaued. Cells were then re-energized with 0.5% glucose and the establishment of a membrane potential was measured as an increase in fluorescence until it plateaued. The membrane potential was then disrupted by the addition of the proton ionophore (CCCP). Data presented is representative of two experiments. For full description, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0183457#sec002" target="_blank">Materials and Methods</a>.</p