Nosocomial Clones Research Articles

Top-Down Genomic Surveillance Approach To Investigate the Genomic Epidemiology and Antibiotic Resistance Patterns of Enterococcus faecium Detected in Cancer Patients in Arkansas.

Control of hospital-associated Enterococcus faecium infection is a strenuous task due to the difficulty of identifying transmission routes and the persistence of this nosocomial pathogen despite the implementation of infection control measures that have been successful with other important nosocomial pathogens. This study provides a comprehensive analysis of over 100 E. faecium isolates collected from 66 cancer patients at the University of Arkansas for Medical Sciences (UAMS) between June 2018 and May 2019. In the top-down approach used in this study, we employed, in addition to the 106 E. faecium UAMS isolates, a filtered set of 2,167 E. faecium strains from the GenBank database to assess the current population structure of E. faecium species and, consequently, to identify the lineages associated with our clinical isolates. We then evaluated the antibiotic resistance and virulence profiles of hospital-associated strains from the species pool, focusing on antibiotics of last resort, to establish an updated classification of high-risk and multidrug-resistant nosocomial clones. Further investigation of the clinical isolates collected from UAMS patients using whole-genome sequencing analytical methodologies (core genome multilocus sequence typing [cgMLST], core single nucleotide polymorphism [coreSNP] analysis, and phylogenomics), with the addition of patient epidemiological data, revealed a polyclonal outbreak of three sequence types occurring simultaneously in different patient wards. The integration of genomic and epidemiological data collected from the patients increased our understanding of the relationships and transmission dynamics of the E. faecium isolates. Our study provides new insights into genomic surveillance of E. faecium to assist in monitoring and further limiting the spread of multidrug-resistant E. faecium. IMPORTANCE Enterococcus faecium is a member of the gastrointestinal microbiota. Although its virulence is low in healthy, immunocompetent individuals, E. faecium has become the third leading cause of health care-associated infections in the United States. This study provides a comprehensive analysis of over 100 E. faecium isolates collected from cancer patients at the University of Arkansas for Medical Sciences (UAMS). We employed a top-down analytical approach (from population genomics to molecular biology) to classify our clinical isolates into their genetic lineages and thoroughly evaluate their antibiotic resistance and virulence profiles. The addition of patient epidemiological data to the whole-genome sequencing analytical methodologies performed in the study allowed us to increase our understanding of the relationships and transmission dynamics of the E. faecium isolates. This study provides new insights into genomic surveillance of E. faecium to help monitor and further limit the spread of multidrug-resistant E. faecium.

Emergence of blaSHV-12 and qnrS1 encoded on IncX3 plasmids: Changing epidemiology of extended-spectrum ß-lactamases among Enterobacterales isolated from broilers

The occurrence of extended-spectrum ß-lactamase (ESBL) producing Enterobacterales in broilers represents a risk to public health because of the possibility of transmission of ESBL-producers and/or blaESBL genes via the food chain or within settings where human-animal interfaces exist. This study assessed the occurrence of ESBL-producers among faecal samples of broilers at slaughter. Isolates were characterised by multilocus sequence typing (MLST), antimicrobial susceptibility testing (AST), and whole genome sequencing (WGS). The flock prevalence, determined by sampling crates of 100 poultry flocks, was 21%. The predominant blaESBL gene was blaSHV-12 identified in 92% of the isolates. A variety of E. coli and K. pneumoniae sequence types (STs) were identified, including extraintestinal pathogenic (ExPEC) E. coli ST38, avian pathogenic E. coli (APEC) ST10, ST93, ST117 and ST155, and nosocomial outbreak clone K. pneumoniae ST20. WGS characterized a subset of 15 isolates, including six Escherichia coli, four Klebsiella pneumoniae, one K. grimontii, one K. michiganensis, one K. variicola, and one Atlantibacter subterranea. Fourteen isolates carried identical or closely related 46338-54929 bp IncX3 plasmids encoding blaSHV-12 and qnrS1. One E. coli isolate carried the 46338 bp IncX3 plasmid integrated chromosomally into ydbD. The blaSHV-12 gene has replaced the previously predominant blaCTX-M-1 in ESBL-producing Enterobacterales from broilers in Switzerland. Broilers may play a role in the dissemination of blaSHV-12 and qnrS1 associated with epidemic IncX3 plasmids, representing a risk to human and animal health.

First Belgian Report of Ertapenem Resistance in an ST11 Klebsiella Pneumoniae Strain Isolated from a Dog Carrying blaSCO-1 and blaDHA-1 Combined with Permeability Defects.

Klebsiella pneumoniae of sequence type (ST) 11 is a hyper-epidemic nosocomial clone, which is spreading worldwide among humans and emerging in pets. This is the first report, to the best of our knowledge, of multidrug-resistant (MDR) K. pneumoniae ST11 carrying blaSCO-1 and blaDHA-1, isolated from a four-month-old dog in Belgium. Antimicrobial susceptibility testing (AST) of the isolate, performed via broth microdilution following the European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines, revealed resistance to eight different classes of antimicrobials, including carbapenems, in particular ertapenem, third-generation cephalosporins and fluoroquinolones. A hybrid approach, combining long- and short-read sequencing, was employed for in silico plasmid characterization, multi-locus sequence typing (MLST) and the identification and localization of antimicrobial resistance (AMR) and virulence-associated genes. Three plasmids were reconstructed from the whole-genome sequence (WGS) data: the conjugative IncFIB(K), the non-mobilizable IncR and the mobilizable but unconjugative ColRNAI. The IncFIB(K) plasmid carried the blaSCO-1 gene, whereas IncR carried blaDHA-1, both alongside several other antimicrobial resistance genes (ARGs). No virulence genes could be detected. Here, we suggest that the resistance to ertapenem associated with susceptibility to imipenem and meropenem in K. pneumoniae could be related to the presence of blaSCO-1 and blaDHA-1, combined with permeability defects caused by point mutations in an outer membrane porin (OmpK37). The presence of the blaSCO-1 gene on a conjugative IncFIB(K) plasmid is worrisome as it can increase the risk of transmission to humans, to animals and to the environment.

Arm race among closely-related carbapenem-resistant Klebsiella pneumoniae clones

Multiple carbapenem-resistant Klebsiella pneumoniae (CRKP) clones typically co-exist in hospital wards, but often certain clones will dominate. The factors driving this dominance are largely unclear. This study began from a genomic epidemiology analysis and followed by multiple approaches to identify the potential mechanisms driving the successful spread of a dominant clone. 638 patients in a 50-bed ICU were screened. 171 (26.8%) and 21 had CRKP from swabs and clinical specimens, respectively. Many (39.8% of those with ≥7-day ICU stay) acquired CRKP. After removing 18 unable to recover, 174 CRKP isolates were genome sequenced and belonged to six sequence types, with ST11 being the most prevalent (n = 154, 88.5%) and most (n = 169, 97.1%) carrying blaKPC-2. The 154 ST11 isolates belonged to 7 clones, with one (clone 1, KL64 capsular type) being dominant (n = 130, 84.4%). Clone 1 and the second-most common clone (clone 2, KL64, n = 15, 9.7%) emerged simultaneously, which was also detected by genome-based dating. Clone 1 exhibited decreased biofilm formation, shorter environment survival, and attenuated virulence. In murine gut, clone 1 outcompeted clone 2. Transcriptomic analysis showed significant upregulation of the ethanolamine operon in clone 1 when competing with clone 2. Clone 1 exhibited increased utilization of ethanolamine as a nitrogen source. This highlights that reduced virulence and enhanced ability to utilize ethanolamine may promote the success of nosocomial multidrug-resistant clones.

Methicillin-resistant Staphylococcus aureus transmission and hospital-acquired bacteremia in a neonatal intensive care unit in Greece

BackgroundStaphylococcus aureus is a common pathogen causing hospital acquired infections (HAIs) in neonates. In this study, the epidemiology of methicillin-resistant S. aureus (MRSA) colonization and infections in a 30-bed, level III university-affiliated neonatal intensive care unit (NICU) located in a children's hospital was retrospectively investigated for the period 2014–2018. MethodsGenes encoding Panton-Valentine Leukocidin (lukS/lukF-PV, PVL), toxic shock syndrome toxin (tst), exfoliative toxins (eta, etb), and the resistance genes mecA, mecC and fusB, were defined in 46 representative strains by PCRs. Relatedness of strains was assessed by MLST. ResultsOf 1538 neonates, 77 (5%) had a positive culture for MRSA (23/77 were NICU-acquired and 54/77 imported cases). Four MRSA bacteremias occurred. Most isolates were multi-resistant. One major clone was identified, ST225, among 40 tested neonatal strains (23/40, 58%). Of these, 14/23 were imported from the same maternity hospital (MH). Another clone, ST217, was predominant (4/6) among health care workers (HCWs), found colonized. Four isolates classified as ST80 were PVL-positive. Additional four strains carried tst (10%), belonging to ST30 and ST225 (two strains each), and two etb. The implicated MH was notified for the problem, decolonization treatment was successfully performed in HCWs and neonates. Strengthening of infection control measures with emphasis on hand hygiene was applied. ConclusionsUncovering reservoirs for on-going MRSA transmission in NICUs has proved challenging. Well known nosocomial MRSA clones are being constantly introduced and transmitted via MHs and HCWs. Effective infection prevention and control requires constant vigilance.

Emergence of livestock-associated MRSA isolated from cystic fibrosis patients: Result of a Belgian national survey

BackgroundThis study aims to determine the prevalence and characteristics of Staphylococcus aureus in Belgian cystic fibrosis (CF) patients. MethodsNon-duplicate respiratory samples from 510 CF-patients (2012−2013) were examined. One isolate per patient was analysed unless different phenotypes were recovered. Isolates were investigated for mecA/mecC, toxins presence, spa-typing, MLST and SCCmec-typing. Potential livestock-associated (LA) isolates were examined for their immune-evasion-cluster (IEC) genes. ResultsS. aureus (n = 380), including 41 small-colony variants (SCVs), were isolated from 66.7% patients. The prevalence of methicillin-resistant S. aureus (MRSA) colonization was 4.9%. Two MRSA isolates carried toxic shock syndrome toxin 1 (TSST-1). Most MRSA (65%) belonged to two nosocomial epidemic clones (CC5, CC8) widespread in Belgium. Methicillin susceptible S. aureus (MSSA) showed great genetic diversity. Five of 33 isolates belonging to potential LA-lineages were IEC negative, including three methicillin-resistant isolates, suggesting an animal origin. ConclusionsThe MRSA-prevalence in Belgian CF-patients remained constant (2001−2013), but SCV-prevalence increased. Most MRSA belonged to health-care-associated clones. Three patients carrying LA-MRSA were found, requiring further investigation to determine the risk factors for LA-MRSA acquisition.

Clinical Implications of Genomic Adaptation and Evolution of Carbapenem-Resistant Klebsiella pneumoniae.

Klebsiella pneumoniae poses a major challenge to healthcare worldwide as an important cause of multidrug-resistant infections. Nosocomial clones, including epidemic sequence type 258 (ST258), have shown an affinity for acquiring and disseminating resistance plasmids, particularly variants of the K. pneumoniae carbapenemase. By comparison, the resurgence of severe community-associated K. pneumoniae infections has led to increased recognition of hypervirulent strains belonging to the K1 and K2 capsular serotypes, predominantly in eastern Asia. Genomic and functional studies suggest that a variety of virulence and immune evasive factors contribute to the success of nosocomial and community-associated clonal lineages, aided by mechanisms of genetic plasticity that contribute to uptake of genes associated with antimicrobial resistance and pathogenicity. While there currently appears to be limited overlap between resistant and hypervirulent lineages, specific bacterial and host factors contributing to the emergence of dominant clones remain incompletely understood. This review summarizes recent advances in our understanding of the molecular epidemiology, virulence potential, and host-pathogen interactions of K. pneumoniae.

The basic mechanisms of the formation of epidemically significant nosocomial bacterial clones

Russian and foreign research on specific characteristics of bacteria that can cause infections associated with the provision of health care, the molecular genetic markers of these bacteria, and the basic mechanisms of the formation of epidemiologically significant nosocomial bacterial clones are reviewed in the present article. It is shown that the main characteristics of nosocomial clones, such as resistance to multiple antimicrobial drugs, genome plasticity, the ability of pathogens to form biofilms, and the presence of virulence factors, are apparently common for all bacterial species. Microevolution processes, including mutations, genomic rearrangements, and horizontal gene transfer, underlie the formation of nosocomial bacterial clones. These processes provide bacteria with selective advantages under certain conditions and thus enable the emergence of epidemics. Keywords: healthcare-associated

Characteristics of the community-genotype sequence type 72 methicillin-resistant Staphylococcus aureus isolates that underlie their persistence in hospitals.

Panton-Valentine leukocidin-negative methicillin-resistant Staphylococcus aureus (MRSA) clone ST72, known as a major community-associated MRSA in Korea, has emerged as an important pathogen in hospitals. To understand bacterial properties that underlie transformation of this clone into a nosocomial pathogen, we compared characteristics of the community-genotype ST72 MRSA isolates with those of ST5 and ST239 MRSA, which have been predominant nosocomial MRSA clones in Korea. Several genes associated with adhesion and virulence were absent or rarely found in ST72 isolates. Many ST72 isolates (70.1%) belonged to agr group I, but the agr group of other ST72 isolates could not be determined. As indicated by d-hemolysin production, ST72 isolates expressed fully functional agr, whereas agr dysfunction was observed in ST5 and ST239 isolates. In the biofilm formation assay, no upregulation of biofilm-forming activity of ST72 MRSA was detected. However, ST72 isolates demonstrated persistence under hypotonic and desiccating conditions (survival rates 72.3% and 33.9%, respectively), which was similar to characteristics of ST5 or ST239 isolates. ST72- MRSA isolates showed low virulence, but properties of their functional agr system could facilitate their spread in hospitals. In conclusion, tolerance to stressful environments, e.g., hypotonic and dry conditions, may also contribute to survival of the community-associated MRSA clones in healthcare facilities.

Both clades of the epidemic KPC-producing Klebsiella pneumoniae clone ST258 share a modified galactan O-antigen type

Klebsiella pneumoniae ST258 is a globally disseminated, extremely drug resistant, nosocomial clone with limited treatment options. We show that the vast majority of ST258 isolates express modified d-galactan-I lipopolysaccharide O-antigen, termed hereinafter as d-galactan-III.The genetic determinant required for galactan-III synthesis was identified as a distinct operon adjacent to the rfb (wb) locus encoding d-galactan-I synthesis. The three genes within the operon encode predicted glycosyltransferases. Testing an isogenic transformant pair revealed that expression of d-galactan-III, in comparison to d-galactan-I, conferred improved survival in the presence of human serum.Eighty-three percent of the more than 200 ST258 draft genome sequences currently available carries the corresponding operon and hence these isolates are predicted to express galactan-III antigens. A d-galactan-III specific monoclonal antibody (mAb) was shown to bind to extracted LPS from a panel of ST258 isolates. The same mAb confirmed accessibility of galactan-III in surface staining of ST258 irrespective of the distinct capsular antigens expressed by both clades described previously.Based on these data, the galactan-III antigen may represent an attractive target for active and passive immunization approaches against K. pneumoniae ST258.

The molecular epidemiology of incident methicillin-resistant Staphylococcus aureus cases among hospitalized patients in Alberta, Canada: a retrospective cohort study.

BackgroundInfection Prevention and Control (IPC) surveillance for incident methicillin-resistant Staphylococcus aureus (MRSA) in hospitalized patients is performed in a complete provincial surveillance network of all acute care facilities in Alberta, Canada. IPC surveillance is centralized using a web-based data entry platform so that each patient is counted only once. All diagnostic laboratories submit the first clinical MRSA isolate associated with a patient without previous MRSA positive clinical cultures in the preceding year to the Provincial Laboratory for Public Health (ProvLab) for molecular typing. This study will investigate the relationship between the IPC epidemiological classification based on time of detection following admission to hospital (Hospital Acquired and Community Associated) and the matched laboratory MRSA surveillance data using a retrospective cohort study design.MethodsIncident IPC MRSA cases were classified according to IPC epidemiologic definitions. DNA sequencing of the Staphylococcus protein A (spa) gene and pulsed-field gel electrophoresis (PFGE) typing was performed. IPC MRSA surveillance data were matched to the ProvLab molecular surveillance data. Univariate comparisons of proportions were performed for categorical variables and the Student’s t test for continuous variables.ResultsMRSA molecular typing data were available for matching for 46.7 % (2248/4818) of incident IPC cases. There was agreement in definitions for traditional nosocomial clones (USA100/CMRSA2) with Hospital Acquired (HA)-MRSA (65.1 % of all IPC HA-MRSA) and traditional community clones (USA400/CMRSA7 and USA300/CMRSA10) with Community Acquired (CA)-MRSA (62.4 % of CA-MRSA). However, we observed discordance for both traditional nosocomial/CA-MRSA (30.4 % of CA-MRSA) and for traditional community/HA-MRSA (26.9 % of HA-MRSA).ConclusionsWe note agreement between traditional nosocomial clones and HA-MRSA, and traditional community clones and CA-MRSA. However, approximately one-quarter of HA-MRSA are those of traditional community clones while approximately one-third of CA-MRSA are those of traditional nosocomial clones. Collaborative provincial MRSA surveillance is important as the distinction between IPC case attribution in acute care settings and the historical definitions of MRSA clones as community- or healthcare-associated have blurred.

WS19.2 Staphylococcus aureus population from cystic fibrosis patients in Spain: Results from a point-prevalence multicentre study

Objectives To characterize Staphylococcus aureus isolates colonising the lung of Spanish cystic fibrosis (CF) patients. Methods A prospective, multicentre study was performed (March-Nov. 2013) covering 341 CF-patients (180 adults, 161 children) from 17 hospitals. Respiratory samples (one per patient) and clinical and microbiological data were sent to our centre. S. aureus isolates were identified (MALDI-TOF MS) and antibiotic susceptibility (MicroScan) determined. Genetic relationships between the isolates were assessed by PFGE- Sma I, spa -typing and MLST, including a concatenate analysis of the alleles. SCC mec and agr type were characterized in methicillin-resistant S. aureus (MRSA). Results A total of 206 S. aureus isolates (60%) were obtained in 97 adults and 109 children. Prevalence in the different CF-Units varied between 13.3–86.6%. Pseudomonas aeruginosa co-colonisation was observed in 36 patients. Twenty-eight MRSA (13.7%) were detected in 11 Units, corresponding to SCCmec-IVc (22/28), SCCmec-IVa (3/28), and SCCmec-V (3/28). Eighty-seven isolates displaying unrelated PFGE were further analysed, showing a high genetic diversity by spa (the most frequent were CC002, CC012) and MLST (CC5, CC8, CC30, and CC45) typing. Three isolates grouped into the ST398, linked to MRSA carriage in pigs. Glycopeptide resistance was absent, whereas a cfr -mediated-linezolid resistant MRSA isolate was detected. Conclusion Overall prevalence of S. aureus in the Spanish CF-population was 60%, being 13.7% of the isolates MRSA. Genetic relationship between isolates was scarce although some widespread nosocomial and community clones were detected, including ST398 from animal origin.

Molecular epidemiology of community-onset methicillin-resistant Staphylococcus aureus infections in Israel.

Data on community-associated (CA) methicillin-resistant Staphylococcus aureus (MRSA) in Israel are scarce. The objective of this study was to characterize the major CA-MRSA clones in Israel. All clinical MRSA isolates detected in the community during a period of 2.5years (2011-2013) from individuals insured by a major health maintenance organization in Israel were collected, with additional data from medical records. Antibiotic susceptibility patterns and staphylococcal chromosomal cassette mec (SCCmec) typing were determined. SCCmec IV and V isolates were further typed by pulsed-field gel electrophoresis (PFGE), spa typing, and detection of a panel of toxin genes. MRSA were detected in 280 patients, mostly from skin infections. Patients with SCCmec IV (n = 120, 43%) were younger (p < 0.0001) and reported less contact with healthcare facilities. Almost all isolates were trimethoprim-sulfamethoxazole susceptible (98%). spa-CC032, a typical nosocomial MRSA clone, accounted for 28% of SCCmec IV. The two major CA-MRSA clones were t008 USA300 (13%) and t991 (10%); t991 was isolated mainly from children (75%), was Panton-Valentine leukocidin (PVL) negative but eta-positive, and was typically susceptible to most antibiotic groups. PVL-positive strains (n = 31) included mainly USA300 (52%) and t019 (13%). While multiple genetic lineages were evident among community-onset MRSA in Israel, approximately 20% are typical CA-MRSA clones, mainly USA300 and a local clone, t991.

Molecular characteristics of vancomycin-resistant Enterococcus faecium from a tertiary care hospital in Chengdu, China

The aim of this study was to characterize vancomycin-resistant Enterococcus faecium (VREfm) isolates phenotypically and molecularly, and investigate associations between the virulence factors enterococcal surface protein (esp), hyaluronidase (hyl), and collagen adhesin (acm) and colonization/infection. A total of 126 E. faecium [66 VREfm and 60 vancomycin-susceptible (VSEfm)] were collected in West China Hospital. Nine E. faecium isolates (7 VREfm and 2 VSEfm) were selected at random for comparative study in a large region from China. Minimum inhibitory concentrations (MICs) were measured by Etest and agar dilution, vancomycin resistance genes (vanA, vanB, and vanC) and virulence genes (esp, acm, and hyl) were detected by polymerase chain reaction (PCR). Thirty-four VREfm underwent repetitive sequence-based PCR (rep-PCR) and multi-locus sequence typing (MLST). One linezolid-resistant isolate (MIC = 8 μg/ml) was found; none were tigecycline resistant. All 73 VREfm (28 infective strains and 45 intestinal colonizers) had the vanA gene and VanA phenotype. Positivity for esp, hyl, and acm in VREfm was 79.5, 46.6, and 86.3%, respectively, which was higher than in VSEfm (54.8, 27.4, and 56.5%, respectively). Among VSEfm, positivity for acm in isolates from pleural or cerebrospinal fluid (84.6%) was higher than that from blood (32.4%). There were 11 rep-PCR types (similarity >95%) and MLST revealed nine sequence types (STs) among the selected isolates. Most VREfm and all VSEfm belonged to clonal complex 17. A new ST was found, with allele sequence (15, 1, 38, 1, 1, 1, 1). In China, most VREfm seem to belong to the classical nosocomial CC17 clone, and many of them have acquired virulence genes, further strengthening a hospital-adapted type.

Extensive Genetic Diversity Identified among Sporadic Methicillin-Resistant Staphylococcus aureus Isolates Recovered in Irish Hospitals between 2000 and 2012

Clonal replacement of predominant nosocomial methicillin-resistant Staphylococcus aureus (MRSA) strains has occurred several times in Ireland during the last 4 decades. However, little is known about sporadically occurring MRSA in Irish hospitals or in other countries. Eighty-eight representative pvl-negative sporadic MRSA isolates recovered in Irish hospitals between 2000 and 2012 were investigated. These yielded unusual pulsed-field gel electrophoresis and antibiogram-resistogram typing patterns distinct from those of the predominant nosocomial MRSA clone, ST22-MRSA-IV, during the study period. Isolates were characterized by spa typing and DNA microarray profiling for multilocus sequence type (MLST) clonal complex (CC) and/or sequence type (ST) and SCCmec type assignment, as well as for detection of virulence and antimicrobial resistance genes. Conventional PCR-based SCCmec subtyping was undertaken when necessary. Extensive diversity was detected, including 38 spa types, 13 MLST-CCs (including 18 STs among 62 isolates assigned to STs), and 25 SCCmec types (including 2 possible novel SCCmec elements and 7 possible novel SCCmec subtypes). Fifty-four MLST-spa-SCCmec type combinations were identified. Overall, 68.5% of isolates were assigned to nosocomial lineages, with ST8-t190-MRSA-IID/IIE±SCCM1 predominating (17.4%), followed by CC779/ST779-t878-MRSA-ψSCCmec-SCC-SCCCRISPR (7.6%) and CC22/ST22-t032-MRSA-IVh (5.4%). Community-associated clones, including CC1-t127/t386/t2279-MRSA-IV, CC59-t216-MRSA-V, CC8-t008-MRSA-IVa, and CC5-t002/t242-MRSA-IV/V, and putative animal-associated clones, including CC130-t12399-MRSA-XI, ST8-t064-MRSA-IVa, ST398-t011-MRSA-IVa, and CC6-t701-MRSA-V, were also identified. In total, 53.3% and 47.8% of isolates harbored genes for resistance to two or more classes of antimicrobial agents and two or more mobile genetic element-encoded virulence-associated factors, respectively. Effective ongoing surveillance of sporadic nosocomial MRSA is warranted for early detection of emerging clones and reservoirs of virulence, resistance, and SCCmec genes.

Contamination of Public Buses with MRSA in Lisbon, Portugal: A Possible Transmission Route of Major MRSA Clones within the Community

In a previous study we have shown that public buses in Oporto, the second largest city in Portugal, were highly contaminated with MRSA. Here we describe the results of a similar study performed in another urban area of Portugal–Lisbon, the capital. Between May 2011 and May 2012, hand touched surfaces of 199 public buses in Lisbon were screened for MRSA contamination. Subsequently, the hands of 575 passengers who frequently use these bus lines were also screened. All hand carriers of MRSA were further screened for nasal carriage. The isolates were characterized by PFGE, staphylococcal cassette chromosome (SCC) mec typing, spa typing, MLST and were tested for the presence of mecA, Panton-Valentine leukocidin and arginine catabolic mobile element genes. MRSA contamination was shown in 72 buses (36.2%). The majority of the isolates belonged to three major clones: Clone A was identified as EMRSA-15 defined by pattern PFGE A, spa types t2357/t747/t025/t379/t910, ST22, and SCCmec IVh (n = 21; 29%). Clone B was the New York/Japan clone characterized by PFGE B-t002/t10682-ST5-II (n = 15; 21%). Clone C included isolates with characteristics of the international community-acquired USA300 or related clones, PFGE C-t008-ST8-IVa/IVc/IVg/IVnt/VI (n = 19; 26%). The first two clones are currently the two major lineages circulating in Portuguese hospitals. The hands of 15 individuals were contaminated with MRSA belonging to the nosocomial clones A or B. Eleven of these individuals were not nasal carriers of MRSA and all but one had travelled by public transportation, namely by bus, prior to sampling. In conclusion, public buses in two major cities in Portugal are often contaminated with MRSA representing clones dominant in hospitals in the particular geographic area. MRSA contamination of public transport and the transfer of the bacteria to the hands of passengers may represent a route through which hospital-acquired MRSA clones may spread to the community.

Methicillin-resistant Staphylococcus aureus carrying SCCmec type II was more frequent than the Brazilian endemic clone as a cause of nosocomial bacteremia

Fifty consecutive MRSA blood isolates were evaluated: 30(60%) carried SCCmec type II (single PFGE clone; sequence type 5 or ST105); 12 (26%), IV; 5 (10%), III; 3 (6%), I. Brazilian endemic clone, carrying SCCmec type III, has been the main nosocomial clone in Brazil; however, this study showed that a clone carrying type II predominated.

Rapid and high-resolution distinction of community-acquired and nosocomial Staphylococcus aureus isolates with identical pulsed-field gel electrophoresis patterns and spa types

Methicillin-resistant Staphylococcus aureus (MRSA) represent a serious threat for public health worldwide. Of particular concern is the emergence of community-acquired MRSA, which is often difficult to distinguish from nosocomial MRSA due to a lack of suitable typing methods for early detection. For example, the USA300 pulsed-field gel electrophoresis (PFGE) pattern includes both the ‘classical’ community-acquired USA300 clone with spa type t008 and an epidemiologically unrelated nosocomial clone with spa type t024. Likewise, spa typing cannot distinguish the classic USA300 from nosocomial MRSA with the spa type t008. Since the fast and high-resolution distinction of these S. aureus types is important for infection prevention and surveillance, we investigated whether multiple-locus variable number tandem repeat fingerprinting (MLVF) can be applied to overcome these limitations. Indeed, MLVF correctly grouped 91 MRSA isolates belonging to the classic USA300 lineage, nosocomial MRSA isolates with the USA300 PFGE profile and spa type t024, and nosocomial MRSA isolates with spa type t008 into 3 distinct clusters. Importantly, several sub-clusters were also identified, reflecting epidemiological relationships between the respective isolates. We conclude that MLVF has the discriminatory power needed to rapidly distinguish very similar community-acquired and nosocomial MRSA isolates and that MLVF-based sub-clustering of isolates is highly useful for epidemiological investigations, outbreak prevention, and control.

Changing Italian nosocomial-community trends and heteroresistance in Staphylococcus aureus from bacteremia and endocarditis

Bloodstream infections due to Staphylococcus aureus (BSI) are serious infections both in hospitals and in the community, possibly leading to infective endocarditis (IE). The use of glycopeptides has been recently challenged by various forms of low-level resistance. This study evaluated the distribution of MSSA and MRSA isolates from BSI and IE in 4 Italian hospitals, their antibiotic susceptibility—focusing on the emergence of hVISA—and genotypic relationships. Our results demonstrate that the epidemiology of MRSA is changing versus different STs possessing features between community-acquired (CA)- and hospital-acquired (HA)-MRSA groups; furthermore, different MSSA isolated from BSI and IE were found, with the same backgrounds of the Italian CA-MRSA. The hVISA phenotype was very frequent (19.5%) and occurred more frequently in isolates from IE and in both the MSSA and MRSA strains. As expected, hVISA were detected in MRSA with vancomycin minimum inhibitory concentrations (MICs) of 1–2 mg/l, frequently associated with the major SCCmec I and II nosocomial clones; this phenotype was also detected in some MSSA strains. The few cases of MR-hVISA infections evaluated in our study demonstrated that 5 out of 9 patients (55%) receiving a glycopeptide, died. Future studies are required to validate these findings in terms of clinical impact.

Surveillance of methicillin-resistant Staphylococcus aureus isolated in Torino (northwest Italy)

Nosocomial infections by methicillin-resistant Staphylococcus aureus (MRSA) are an increasing cause of morbidity and mortality. Recently, a worldwide increase of community-acquired MRSA infections has also been recorded. The purpose of this study was to assess the frequency of MRSA isolation from in- and outpatients admitted to an academic teaching hospital near Torino (northwest Italy) in 1 year and to characterize 90 clinical isolates of MRSA collected in the same period. Antimicrobial susceptibility and the presence in the isolates of the Panton–Valentine leukocidin (PVL) gene were assessed. Molecular epidemiology was performed by SCC mec and capsule typing, and by pulsed-field gel electrophoresis. The global proportion of MRSA isolated was 33.1%. Characterization performed on 90 MRSA revealed a high percentage of resistance to ciprofloxacin and erythromycin, and the presence of the PVL gene in one strain only. Most of the MRSA strains circulating in the Torino district belonged to SCC mec types II and I, and the 67.6% resulted positive for the cap 5 gene. The pulsotype analysis permitted to observe a clonal heterogeneity of the isolates and a higher similarity in relation to singular mec types; only few nosocomial clones could account for a local outbreak of a sporadic isolate.