Detection of <i>mecC-</i>Positive <i>Staphylococcus aureus</i> (CC130-MRSA-XI) in Diseased European Hedgehogs (<i>Erinaceus europaeus</i>) in Sweden

<div><p>Recently, a novel <i>mec</i> gene conferring beta-lactam resistance in <i>Staphylococcus aureus</i> has been discovered. This gene, <i>mecC</i>, is situated on a SCC<i>mec</i> XI element that has to date been identified in clonal complexes 49, 130, 425, 599 and 1943. Some of the currently known isolates have been identified from animals. This, and observations of <i>mecA</i> alleles that do not confer beta-lactam resistance, indicate that <i>mec</i> genes might have a reservoir in <i>Staphylococcus</i> species from animals. Thus it is important also to screen wildlife isolates for <i>mec</i> genes. Here, we describe <i>mecC</i>-positive <i>Staphylococcus aureus</i> (ST130-MRSA-XI) and the lesions related to the infection in two diseased free-ranging European hedgehogs (<i>Erinaceus europaeus</i>). One was found dead in 2003 in central Sweden, and suffered from <i>S. aureus</i> septicaemia. The other one, found on the island of Gotland in the Baltic Sea in 2011, showed a severe dermatitis and was euthanised. ST130-MRSA-XI isolates were isolated from lesions from both hedgehogs and were essentially identical to previously described isolates from humans. Both isolates carried the complete SCC<i>mec</i> XI element. They lacked the <i>lukF-PV/lukS-PV</i> and <i>lukM/lukF-P83</i> genes, but harboured a gene for an exfoliative toxin homologue previously described from <i>Staphylococcus hyicus</i>, <i>Staphylococcus pseudintermedius</i> and other <i>S. aureus</i> of the CC130 lineage. To the best of our knowledge, these are the first reported cases of CC130-MRSA-XI in hedgehogs. Given that one of the samples was taken as early as 2003, this was the earliest detection of this strain and of <i>mecC</i> in Sweden. This and several other recent observations suggest that CC130 might be a zoonotic lineage of <i>S. aureus</i> and that SCC<i>mec</i> XI/<i>mecC</i> may have originated from animal pathogens.</p></div

Novel primer and probe sequences and PCR conditions.

<p>Novel primer and probe sequences and PCR conditions.</p

The Emergence and Spread of Multiple Livestock-Associated Clonal Complex 398 Methicillin-Resistant and Methicillin-Susceptible <i>Staphylococcus aureus</i> Strains among Animals and Humans in the Republic of Ireland, 2010–2014

<div><p>Clonal complex (CC) 398 methicillin-resistant <i>Staphylococcus aureus</i> (MRSA) and methicillin-susceptible <i>S</i>. <i>aureus</i> (MSSA) are associated with carriage and infection among animals and humans but only a single case of CC398 MRSA has been reported in the Republic of Ireland (ROI). The present study investigated the molecular epidemiology of CC398 MRSA (<i>n</i> = 22) and MSSA (<i>n</i> = 10) from animals and humans in the ROI from 2010–2014. Isolates underwent antimicrobial susceptibility testing, <i>spa</i> typing, DNA microarray profiling and PCR for CC398-associated resistance genes. All MRSA underwent SCC<i>mec</i> IV or V subtyping. Four distinct CC398-MRSA incidents were identified from (i) a man in a nursing home (<i>spa</i> type t011-SCC<i>mec</i> IVa, immune evasion complex (IEC) negative), (ii) a horse and veterinarian who had recently travelled to Belgium (t011-IVa, IEC positive), (iii) pigs (<i>n</i> = 9) and farm workers (<i>n</i> = 9) on two farms, one which had been restocked with German gilts and the other which was a finisher farm (t034-V_T, IEC negative, 3/9 pigs; t011- V_T, IEC negative, 6/9 pigs & 9/9 farm workers), and (iv) a child who had worked on a pig farm in the UK (t034-V_T, IEC negative). Isolates also carried different combinations of multiple resistance genes including <i>erm</i>(A), <i>erm</i>(B), <i>tet</i>(K), <i>tet</i>(M) & <i>tet</i>(L), <i>fexA</i>, <i>spc</i>, <i>dfrG</i>, <i>dfrK aacA-aphD</i> and <i>aadD</i> further highlighting the presence of multiple CC398-MRSA strains. CC398 MSSA were recovered from pigs (<i>n</i> = 8) and humans (<i>n</i> = 2). CC398 MSSA transmission was identified among pigs but zoonotic transmission was not detected with animal and human isolates exhibiting clade-specific traits. This study highlights the importation and zoonotic spread of CC398 MRSA in the ROI and the spread of CC398 MSSA among pigs. Increased surveillance is warranted to prevent further CC398 MRSA importation and spread in a country that was considered CC398 MRSA free.</p></div

Epidemiological, phenotypic and genotypic characteristics of CC398 methicillin-resistant and methicillin-susceptible <i>S</i>. <i>aureus</i> (MRSA and MSSA) identified in the Republic of Ireland among animals and humans.

<p>Epidemiological, phenotypic and genotypic characteristics of CC398 methicillin-resistant and methicillin-susceptible <i>S</i>. <i>aureus</i> (MRSA and MSSA) identified in the Republic of Ireland among animals and humans.</p

Skin lesions in a hedgehog with MRSA dermatitis (V5406/11).

<p>a) H&E staining of an ulceration with necrosis of surface epithelium, thick serocellular crusts and irregular epidermal hyperplasia. b) H&E staining of an inflammatory infiltrate of lymphocytes, macrophages and few neutrophils in the superficial dermis. c) H&E staining of a superficial dermis subjacent to ulceration. Necrosis, cell debris, severe infiltrate of neutrophils and mononuclear cells and haemorrhages.</p

Image_1_Intra-Hospital, Inter-Hospital and Intercontinental Spread of ST78 MRSA From Two Neonatal Intensive Care Unit Outbreaks Established Using Whole-Genome Sequencing.PDF

<p>From 2009 to 2011 [transmission period (TP) 1] and 2014 to 2017 (TP2), two outbreaks involving community-associated clonal complex (CC) 88-MRSA spa types t186 and t786, respectively, occurred in the Neonatal Intensive Care Unit (NICU) of an Irish hospital (H1). This study investigated the relatedness of these isolates, their relationship to other CC88 MRSA from Ireland and their likely geographic origin, using whole-genome sequencing (WGS). All 28 CC88-MRSA isolates identified at the Irish National MRSA Reference Laboratory between 2009 and 2017 were investigated including 20 H1 patient isolates, two H1 isolates recovered from a single healthcare worker (HCW) 2 years apart, three patient isolates from a second hospital (H2) and one patient isolate from each of three different hospitals (H3, H4, and H5). All isolates underwent DNA microarray profiling. Thirteen international isolates with similar microarray profiles to at least one Irish isolate were selected from an extensive global database. All isolates underwent Illumina MiSeq WGS. The majority of Irish isolates (25/28; all H1 isolates, two H2 isolates and the H3 isolate) were identified as ST78-MRSA-IVa and formed a large cluster, exhibiting 1–71 pairwise allelic differences, in a whole-genome MLST-based minimum spanning tree (MST) involving all Irish isolates. A H1/H2, H1/H3, and H1 HCW/patient isolate pair each exhibited one allelic difference. The TP2 isolates were characterised by a different spa type and the loss of hsdS. The three remaining Irish isolates (from H2, H4, and H5) were identified as ST88-MRSA-IVa and dispersed at the opposite end of the MST, exhibiting 81–211 pairwise allelic differences. Core-genome MLST and sequence-based plasmid analysis revealed the recent shared ancestry of Irish and Australian ST78-MRSA-IVa, and of Irish and French/Egyptian ST88-MRSA-IVa. This study revealed the homogeneity of isolates recovered during two NICU outbreaks (despite spa type and hsdS carriage variances), HCW involvement in the outbreak transmission chain and the strain's spread to two other Irish hospitals. The outbreak strain, CC88/ST78-MRSA-IVa, was likely imported from Australia, where it is prevalent. CC88/ST88-MRSA-IVa was also identified in Irish hospitals and was likely imported from Africa, where it is predominant, and/or a country with a large population of African descent.</p

The recent emergence in hospitals of multidrug-resistant community-associated sequence type 1 and <i>spa</i> type t127 methicillin-resistant <i>Staphylococcus aureus</i> investigated by whole-genome sequencing: Implications for screening

<div><p>Community-associated <i>spa</i> type t127/t922 methicillin-resistant <i>Staphylococcus aureus</i> (MRSA) prevalence increased from 1%-7% in Ireland between 2010–2015. This study tracked the spread of 89 such isolates from June 2013-June 2016. These included 78 healthcare-associated and 11 community associated-MRSA isolates from a prolonged hospital outbreak (H1) (<i>n</i> = 46), 16 other hospitals (<i>n</i> = 28), four other healthcare facilities (<i>n</i> = 4) and community-associated sources (<i>n</i> = 11). Isolates underwent antimicrobial susceptibility testing, DNA microarray profiling and whole-genome sequencing. Minimum spanning trees were generated following core-genome multilocus sequence typing and pairwise single nucleotide variation (SNV) analysis was performed. All isolates were sequence type 1 MRSA staphylococcal cassette chromosome <i>mec</i> type IV (ST1-MRSA-IV) and 76/89 were multidrug-resistant. Fifty isolates, including 40/46 from H1, were high-level mupirocin-resistant, carrying a conjugative 39 kb <i>iles2</i>-encoding plasmid. Two closely related ST1-MRSA-IV strains (I and II) and multiple sporadic strains were identified. Strain I isolates (57/89), including 43/46 H1 and all high-level mupirocin-resistant isolates, exhibited ≤80 SNVs. Two strain I isolates from separate H1 healthcare workers differed from other H1/strain I isolates by 7–47 and 12–53 SNVs, respectively, indicating healthcare worker involvement in this outbreak. Strain II isolates (19/89), including the remaining H1 isolates, exhibited ≤127 SNVs. For each strain, the pairwise SNVs exhibited by healthcare-associated and community-associated isolates indicated recent transmission of ST1-MRSA-IV within and between multiple hospitals, healthcare facilities and communities in Ireland. Given the interchange between healthcare-associated and community-associated isolates in hospitals, the risk factors that inform screening for MRSA require revision.</p></div

Comparative Genotypes, Staphylococcal Cassette Chromosome mec (SCCmec) Genes and Antimicrobial Resistance amongst <i>Staphylococcus epidermidis</i> and <i>Staphylococcus haemolyticus</i> Isolates from Infections in Humans and Companion Animals

<div><p>This study compares the characteristics of <i>Staphylococcus epidermidis</i> (SE) and <i>Staphylococcus haemolyticus</i> (SH) isolates from epidemiologically unrelated infections in humans (Hu) (28 SE-Hu; 8 SH-Hu) and companion animals (CpA) (12 SE-CpA; 13 SH-CpA). All isolates underwent antimicrobial susceptibility testing, multilocus sequence typing and DNA microarray profiling to detect antimicrobial resistance and SCC<i>mec</i>-associated genes. All methicillin-resistant (MR) isolates (33/40 SE, 20/21 SH) underwent <i>dru</i> and <i>mecA</i> allele typing. Isolates were predominantly assigned to sequence types (STs) within a single clonal complex (CC2, SE, 84.8%; CC1, SH, 95.2%). SCC<i>mec</i> IV predominated among MRSE with ST2-MRSE-IVc common to both Hu (40.9%) and CpA (54.5%). Identical <i>mecA</i> alleles and nontypeable <i>dru</i> types (dts) were identified in one ST2-MRSE-IVc Hu and CpA isolate, however, all <i>mecA</i> alleles and 2/4 dts detected among 18 ST2-MRSE-IVc isolates were closely related, sharing >96.5% DNA sequence homology. Although only one ST-SCC<i>mec</i> type combination (ST1 with a non-typeable [NT] SCC<i>mec</i> NT9 [class C <i>mec</i> and <i>ccrB4</i>]) was common to four MRSH-Hu and one MRSH-CpA, all MRSH isolates were closely related based on similar STs, SCC<i>mec</i> genes (V/V_T or components thereof), <i>mecA</i> alleles and dts. Overall, 39.6% of MR isolates harbored NT SCC<i>mec</i> elements, and ACME was more common amongst MRSE and CpA isolates. Multidrug resistance (MDR) was detected among 96.7% of isolates but they differed in the prevalence of specific macrolide, aminoglycoside and trimethoprim resistance genes amongst SE and SH isolates. Ciprofloxacin, rifampicin, chloramphenicol [<i>fexA</i>, <i>cat-pC221</i>], tetracycline [<i>tet</i>(K)], aminoglycosides [<i>aadD</i>, <i>aphA3</i>] and fusidic acid [<i>fusB</i>] resistance was significantly more common amongst CpA isolates. SE and SH isolates causing infections in Hu and CpA hosts belong predominantly to STs within a single lineage, harboring similar but variable SCC<i>mec</i> genes, <i>mecA</i> alleles and dts. Host and staphylococcal species-specific characteristics were identified in relation to antimicrobial resistance genes and phenotypes, SCC<i>mec</i> and ACME.</p></div