Dissemination Of Methicillin-resistant Staphylococcus Aureus Research Articles

Exploring the virulence potential of immune evasion cluster genes in methicillin-resistant Staphylococcus aureus from cancer patients

Methicillin-resistant Staphylococcus aureus (MRSA) is accountable for a plethora of infections, ranging from minor cutaneous manifestations to grave metastatic conditions. The dissemination of MRSA among cancer patients poses a substantial public health hazard on a global scale. This study explores the association between MRSA and bacteriophage-encoded immune evasion cluster (IEC) genes. This investigation employed a total of 168 pathogenic MRSA collected from 38 cancer and 130 non-cancer patients. A cefoxitin disc diffusion method followed by PCR analysis was used to identify the mecA gene. In this study, we employed singleplex and multiplexed PCR techniques to detect specific IEC genes. No association (p = 0.98) was observed between the sex and age of patients and MRSA isolates. However, MRSA isolates demonstrated a notable association (p = 0.01) with pus samples in non-cancer patients and skin swabs in cancer patients. The resistance profiles of MRSA strains from cancer and non-cancer patients did not show significant differences (p > 0.05). Notably, the sea gene was found to be more prevalent in MRSA isolates from cancer patients, displaying a significant association (p = 0.03). Additionally, this study identified two novel and distinct combinations of IEC types, namely V1 (sea, chp, scn) and V2 (sea, scn). Cancer patients had higher multidrug resistance and toxin gene abundance than non-cancer patients. The identification of two novel IEC patterns underscores the urgent need to control MRSA dissemination in hospitals and monitor emerging clones.

Dynamics of the MRSA Population in a Chilean Hospital: a Phylogenomic Analysis (2000-2016).

The global dissemination of methicillin-resistant Staphylococcus aureus (MRSA) is associated with the emergence and establishment of clones in specific geographic areas. The Chilean-Cordobes clone (ChC) (ST5-SCCmecI) has been the predominant MRSA clone in Chile since its first description in 1998, despite the report of other emerging MRSA clones in recent years. Here, we characterize the evolutionary history of MRSA from 2000 to 2016 in a Chilean tertiary health care center using phylogenomic analyses. We sequenced 469 MRSA isolates collected between 2000 and 2016. We evaluated the temporal trends of the circulating clones and performed a phylogenomic reconstruction to characterize the clonal dynamics. We found a significant increase in the diversity and richness of sequence types (STs; Spearman r = 0.8748, P < 0.0001) with a Shannon diversity index increasing from 0.221 in the year 2000 to 1.33 in 2016, and an effective diversity (Hill number; q = 2) increasing from 1.12 to 2.71. The temporal trend analysis revealed that in the period 2000 to 2003 most of the isolates (94.2%; n = 98) belonged to the ChC clone. However, since then, the frequency of the ChC clone has decreased over time, accounting for 52% of the collection in the 2013 to 2016 period. This decline was accompanied by the rise of two emerging MRSA lineages, ST105-SCCmecII and ST72-SCCmecVI. In conclusion, the ChC clone remains the most frequent MRSA lineage, but this lineage is gradually being replaced by several emerging clones, the most important of which is clone ST105-SCCmecII. To the best of our knowledge, this is the largest study of MRSA clonal dynamics performed in South America. IMPORTANCE Methicillin-resistant Staphylococcus aureus (MRSA) is a major public health pathogen that disseminates through the emergence of successful dominant clones in specific geographic regions. Knowledge of the dissemination and molecular epidemiology of MRSA in Latin America is scarce and is largely based on small studies or more limited typing techniques that lack the resolution to represent an accurate description of the genomic landscape. We used whole-genome sequencing to study 469 MRSA isolates collected between 2000 and 2016 in Chile providing the largest and most detailed study of clonal dynamics of MRSA in South America to date. We found a significant increase in the diversity of MRSA clones circulating over the 17-year study period. Additionally, we describe the emergence of two novel clones (ST105-SCCmecII and ST72-SCCmecVI), which have been gradually increasing in frequency over time. Our results drastically improve our understanding of the dissemination and update our knowledge about MRSA in Latin America.

Continuous medical education activities; Answers to Case No. 5: Infection control for MRSA transmission

Methicillin-resistant Staphylococcus aureus (MRSA) is still recognized as one of the most important nosocomial pathogens. These isolates are usually resistant to all currently available β-lactam antibiotics (penicillins, cephalosporins and carbapenems). Vancomycin has historically been the drug of choice and sometimes the last resort for the treatment of serious MRSA infections, providing empirical coverage and definitive therapy. However, its increased use has now become questionable. Moreover, its increased use has already led to emergence of vancomycin-intermediate S. aureus (VISA) as well as vancomycin-resistant S. aureus (VRSA) in certain parts of the world. In the early 1990s, MRSA was reported to account for 20 – 25 % of S. aureus isolates in hospitalized, worldwide. By the middle of the current decade, many hospitals experienced MRSA percentages in the range of 50-70% of total S. aureus isolates from clinical cultures. Recent studies have found that an increasing proportion of hospital-onset invasive MRSA infections are caused by community strains. The clinical scenario has been more dramatic by MRSA colonization which increases the risk of infection, and infecting strains match colonizing strains in as many as 50–80% of cases. Methicillin-resistant Staphylococcus aureus may persist within the hospital environment for a long time, complicating attempts of eradication. Besides, colonization is not static, as strains have been found to evolve and even to be replaced within the same host. Poor infection control measures as well as continues and indiscriminate use of antibiotics have resulted in this huge problem of acquisition and dissemination of MRSA.

Emergence of HA-MRSA Associated Staphylococcal Cassette Chromosome mec (SCCmec) Elements Spillover to LA-MRSA

Methicillin-resistant Staphylococcus aureus (MRSA) once confined to hospitals (HA-MRSA), in the last two decades, have emerged in the community in people with no exposure to hospital and also in livestock animals, referred to as community-associated (CA-) and livestock-associated (LA-) MRSA, respectively. Prevention of the dissemination of MRSA requires better knowledge of its distribution and transmission. The study aims to investigate samples from different animal farms and veterinary clinics for identification and typing of MRSA circulating among different farm animals. Phenotypic and genetic characteristics were evaluated based on morpho-cultural and biochemical properties and PCR amplification of nuc genes. Methicillin resistance was assessed on ORSAB media and PCR amplification of methicillin resistance mecA gene. SCCmec typing was performed by multiplex PCR to assess the epidemiological relationship between the isolates. Of the 406 samples, 72 confirmed the presence of S. aureus. Among S. aureus isolates, based on PCR amplification of mecA gene, 15 were identified as LA-MRSA. Majority of the LA-MRSA were found to be SCCmec type III with two isolates each from cattle and pig showed presence of both type I and III. The finding suggests the spillover of the SCCmec type III into livestock and emergence of new clones possessing both type I and III elements.

Antimicrobial Resistance and Virulence of Methicillin-Resistant Staphylococcus aureus from Human, Chicken and Environmental Samples within Live Bird Markets in Three Nigerian Cities.

Background: Methicillin-resistant Staphylococcus aureus (MRSA) has emerged as a major threat to public health. This study investigated the occurrence of MRSA in humans, chickens, chicken meat and environmental samples within poultry farms and live bird markets in southwestern Nigeria. Methods: MRSA were isolated using selective culture and tested for antimicrobial susceptibility by broth microdilution. Selected isolates were characterized by whole genome sequencing (WGS). From WGS data, spa, dru, multilocus sequence typing (MLST) and SCCmec types, but also virulence and antimicrobial resistance genes, were identified. Results: Fifty-six MRSA isolates were detected in 734 samples. They showed resistance to β-lactams (100%), tetracycline (60.7%), ciprofloxacin (33.9%), erythromycin (28.6%), gentamicin (32.1%), and trimethoprim/sulfamethoxazole (10.7%). All 30 isolates investigated by WGS carried mecA, dfrG, and tet(38) genes. Other resistance genes detected were blaZ (83.3%), fosB (73.3%), tet(K) (60.0%), aacA-aphD (36.6%), aphA3 (33.3%), msr(A) (30.0%), mph(C) (30.0%), dfrS1 (3.3%), and sat4 (3.3%). Seven spa types (t091, t314, t657, t1476, t2331, t4690 and t12236), four known (dt9aw, dt10ao, dt10cj, and dt11a) and two novel (dt10dr and dt11dw) dru types, as well as five sequence types (ST8, ST121, ST152, ST772 and ST789) were found among the MRSA isolates. All ST121 isolates carried an SCCmec type IV cassette and were not dru-typeable. ST152 and ST121 were found only in specific sample categories within defined locations, while ST8 and ST772 were distributed across most sample categories and locations. Three SCCmec types, IVa, V and Vc, were identified. All MRSA isolates possessed virulence genes including aur, clpP, coa, fnbA, esaA, hly, hla, ica, isdA, srtB, sspA, and vWbp, among others. The toxic shock syndrome toxin gene (tst) was not detected in any isolate, whereas the Pantone–Valentine leukocidin genes lukF-PV/lukS-PV were present in all ST121, all ST772, and all but one ST152 isolates. Conclusion: The results of this study (i) showed that chicken meat is contaminated by MRSA and (ii) suggested that live bird markets may serve as focal points for the dissemination of MRSA within the community.

Colonization of Methicillin-Resistant Staphylococcus aureus (MRSA) among Medical Students in Tertiary Institution in Central Malaysia.

Methicillin-resistant Staphylococcus aureus or MRSA infection is virulent and presents with a broad spectrum of severity. Limited regional reports that specifically outlined the potential risk of medical students being part of the dissemination of MRSA in healthcare settings were noted. This study aims to assess the prevalence and contributory factors of colonization of MRSA on neckties, headscarves, and ID badges among medical students in a local medical university in Malaysia. A cross-sectional study was conducted involving 256 medical students. A validated questionnaire was used to collect the data, and sample swabs were collected between July and August 2013 by swabbing neckties, headscarves, or identification badges. The swabs were then streaked onto mannitol salt agar (MSA) and incubated at 37 °C overnight. Out of 433 samples taken, 40 swabs (9.24%) were positive for Staphylococcus aureus. Out of the 40 swabs, five (12.5%) isolates were MRSA (one culture was isolated from the headscarf of a preclinical student, one culture was isolated from the necktie of clinical students, while the remaining three were isolated from identification badges of clinical students. There was no significant association between age, gender, ethnicity, and phase of medical students with the colonization of MRSA (p > 0.05). There was a significant association between knowledge score on hand hygiene practice and phase of medical students. MRSA colonies were present on neckties, headscarves, and identification badges of medical students of all phases. The findings from this study suggest the need for improvement of hand hygiene knowledge and discontinuity of mandatory use of physical ID badges and neckties among medical students.

Characterization of Staphylococcus aureus isolated from tilapia and utensils used in the commerciali- zation of tilapia in the street markets of a semi-arid Brazilian municipality.

The production and commercialization of tilapia (Oreochromis niloticus) is fundamentally important to the semi-arid region of northeastern Brazil. In this region, one of the main forms of commercialization occurs in street markets (fairs). A high incidence of methicillin-resistant Staphylococcus aureus (MRSA) strains was previously detected in different food-related environments in Brazil. However, limited data is available about the presence of MRSA in street markets commercializing tilapias. In this study, we identified Staphylococcus aureus from tilapia and utensils used in the commercialization of tilapia in the street markets of a semi-arid Brazilian municipality and characterized the virulence potential of the isolates by analyzing their &beta;-lactam resistance, intercellular adhesion and enterotoxin genes. The study used samples from the 5 main markets in the city. Phenotypic tests to determine antimicrobial resistance, exopolysaccharide (EPS) production, the potential for biofilm formation and cell surface hydrophobicity were conducted on S. aureus isolates. The presence of antimicrobial resistance genes (mecA and blaZ), potential biofilm production genes (icaA and icaD) and enterotoxin (se) genes was investigated. Coagulase-positive staphylococci (CoPS) were detected in samples from all markets in discordance with the legal limits in force. Twelve isolates were confirmed to be S. aureus. Ten isolates demonstrated multidrug resistance (MDR). All isolates were able to produce EPS and form biofilms. Eight isolates exhibited strong hydrophobicity and six a high potential for biofilm formation. Twelve isolates were positive for mecA, blaZ, icaD and sed. Tilapia marketed in unsuitable conditions may be a vehicle for staphylococcal food poisoning and for the dissemination of MRSA to consumers. Additionally, the ability of the isolates to produce biofilms is an alert to the presence and persistence of these virulent microorganisms on utensils used for the commercial distribution of tilapia.

Clinical and molecular characterization of both methicillin-resistant andsensitive staphylococcus aureus mastitis

This study targeted bovine mastitis as a possible source of livestock-associated methicillin-resistant Staphylococcus aureus (MRSA), to identify clinical signs associated with MRSA- and non-MRSA-associated mastitis. Thirty-eight mastitis cases (68 infected quarters) were investigated. Gram-positive cocci-shaped isolates were selected based on Baird Parker agar growth as well as Gram-stained bacterial smears. Molecular screening for Staphylococcus aureus (S. aureus) yielded 17 isolates, of which five (29.41%) were methicillin resistant. The five isolates were mecA positive, but mecC negative. Multilocus sequence typing (MLST) indicated that sequence type 1 (ST1) was the identified type of all isolates of MRSA. S. aureus-associated cases showed different clinical forms of mastitis, including subclinical, acute, chronic, and gangrenous. Additionally, subclinical mastitis was the only detected condition associated with MRSA, which may represent a potential hidden risk for humans. Phenotypically, isolates of MRSA showed resistance to all of the tested β-lactam antimicrobials, with marked resistance to tetracycline and gentamycin. Based on our knowledge, this is the first report to identify MRSA ST1 in Egypt. Bovine mastitis could be a source for the dissemination of MRSA to humans and other animals. Additionally, while methicillin-resistance may have no effect on the clinical outcome of mastitis, it does affect therapeutic success, particularly when β-lactam antimicrobials are used.

European hedgehogs (Erinaceus europaeus) as a natural reservoir of methicillin-resistant Staphylococcus aureus carrying mecC in Denmark.

ObjectivesA recent study from Sweden showed that European hedgehogs may constitute a reservoir for methicillin-resistant Staphylococcus aureus (MRSA), but this host-parasite relationship remains to be investigated in other countries. In this study, we therefore sought to: 1) determine the dissemination of MRSA in European hedgehogs throughout Denmark; 2) investigate determinants of MRSA carriage in hedgehogs; 3) determine the potential for zoonotic transmission of MRSA from hedgehogs to humans; and 4) characterise the detected MRSA on both a phenotypic and molecular level.MethodsNasal swabs were taken from 188 dead hedgehogs collected by volunteers throughout Denmark to determine the occurrence of MRSA. Additionally, 16 hedgehog rehabilitators were tested for potential zoonotic transmission of MRSA from hedgehogs to humans. The swabs were incubated in tryptic soy broth supplemented with 6.5% NaCl, followed by spread of 10 μl on Brilliance MRSA 2 agar. One presumptive MRSA colony from each plate was subcultured on 5% blood agar. All S. aureus subcultures were verified by a PCR assay detecting mecA, mecC, lukF-PV, scn, and spa, followed by spa typing.ResultsA total of 114 (61%) hedgehogs carried mecC-MRSA, whereas none carried mecA-MRSA. The detected mecC-MRSA belonged to two genetic lineages CC130 (spa-types: t528, t843, t1048, t3256, t3570, t6220, t17133) and CC1943 (spa-types: t978, t2345, t3391, t8835, t16868), 52% of which were spa-type t843 (CC130).The detection rate of mecC-MRSA in the hedgehogs was similar regardless of cause of death, sex, region and habitat type. None of the hedgehog rehabilitators carried MRSA.ConclusionsThis nationwide study confirms a high occurrence of mecC-MRSA in hedgehogs, which could serve as a natural reservoir for this specific type of MRSA. Furthermore, our study did not find signs of zoonotic transmission of mecC-MRSA to hedgehog rehabilitators.

Staphylococcus aureus and bovine mastitis: molecular typing of methicillinresistance and clinical description of infected quarters

This study targeted Staphylococcus aureus (S. aureus)-mastitis of bovine as a possible source of livestock-associated methicillin-resistant Staphylococcus aureus (MRSA), to describe clinical signs of mastitis associated with MRSA and methicillin-sensitive Staphylococcus aureus (MSSA). The study area was the Gharbia and Kafrelsheikh governorates, in the central and northern regions of the Egyptian Delta. Clinical examination of animals was performed to detect clinical mastitis (CM) and clinically normal cases were tested by the California mastitis test (CMT) to identify subclinical mastitis (SCM). Accordingly, 38 mastitis cases (68 infected quarters) were detected. Milk samples were collected aseptically and were cultured on Baird Parker agar. Fifty nine Gram-positive cocci-shaped isolates were selected and preserved. In addition, 33 Staphylococcus spp. isolates originated from bovine mastitis at the same study area were obtained from Animal Health Research Institute (AHRI). The cocci-shaped Gram-positive bacteria and AHRI Staphylococcus spp. isolates were used for molecular identification of S. aureus and MRSA. Molecular screening had yielded 17 S. aureus isolates, from which five isolates (29.41%) were MRSA and 12 isolates (70.59%) were MSSA. The five MRSA isolates were mecA positive, but mecC negative. Multilocus sequence typing (MLST) of the five MRSA isolates indicated that all were sequence type 1 (ST1). S. aureus-associated cases showed different clinical forms of mastitis, including subclinical, acute, chronic, and gangrenous. However, subclinical mastitis was the only detected form associated with MRSA, which may represent a potential hidden risk for humans. Phenotypic antimicrobial resistance pattern of MRSA isolates showed resistance to all of the tested β-lactam antimicrobials, with marked orgaresistance to tetracycline and gentamycin. Based on our knowledge, this is the first report to identify MRSA ST1 in Egypt. Bovine mastitis could be a source for the dissemination of MRSA to humans and other animals. Additionally,while methicillin-resistance may have no effect on the clinical severity of mastitis, it does affect therapeutic success, particularly when β-lactam antimicrobials are used.

Prevalence, Molecular Characterization, and Antimicrobial Susceptibility of Methicillin-Resistant Staphylococcus aureus from Different Origins in Sichuan Province, China, 2007-2015.

Methicillin-resistant Staphylococcus aureus (MRSA) is a major multidrug-resistant (MDR) opportunistic pathogen that is responsible for nosocomial infections worldwide. The emergence of MRSA in food-producing animals has heightened concerns regarding the presence of MRSA in foods having an animal origin. The objectives of this study were to determine the prevalence of MRSA in different sources, including retail food, food-producing animals, and food handlers in Sichuan province. During 2007-2015, S. aureus was isolated from samples having different origins. Susceptibilities of MRSA to a panel of 12 antimicrobial agents were determined according to Clinical and Laboratory Standards Institute (CSLI) procedures. Pulsed-field gel electrophoresis, spa typing, staphylococcal chromosomal cassette (SCC) mec typing, and multilocus sequence typing (MLST) were used to characterize the relationship between these isolates. A total of 756 S. aureus were isolated from 11,067 samples. Of these, 52 isolates were classified as MRSA based on the presence of mecA genes. An antimicrobial susceptibility assay indicated that CHL-CLI-ERY-FOX-OXA-PEN-TET (n = 10) and CHL-CIP-CLI-ERY-FOX-OXA-PEN-TET (n = 8) were the two predominant MDR profiles in the isolates. Using 60% genetic similarity as a cutoff, the 52 MRSA isolates were grouped into 6 clusters having 34 pulsotypes. MLST typing showed seven multilocus sequence types (STs) with ST59 and ST9 being the most common. Six SCCmec types were identified in all MRSA isolates. The MRSA isolates had relatively low prevalence in Sichuan province. Clonal expansion is not involved in the dissemination of MRSA from food origins. Considering the threat of MRSA to public health, further surveillance is required to monitor the prevalence of food-related MRSA.

Nasal carriage of sequence type 22 MRSA and livestock-associated ST398 clones in Tangier, Morocco.

This study aimed to provide data of Staphylococcus aureus and methicillin-resistant Staphylococcus aureus (MRSA) nasal carriage as well as to determine the genetic lineages of this circulating MRSA in the Tangier community. Between 2012 and 2013 two subpopulations consisting of randomly chosen healthy volunteers and outpatients in 11 healthcare facilities were screened. The antibiotic resistance phenotype was determined by disk diffusion. Toxin Panton-Valentin Leukocidin (PVL), toxic shock syndrome toxin-1 gene (tst), and mecA were detected by polymerase chain reaction (PCR). Nasal swabs were obtained from persons with no identified risk factors for MRSA acquisition. MRSA molecular typing was performed by pulsed-field gel electrophoresis (PFGE), staphylococcal chromosomal cassette mec, and Staphylococcus protein A (spa) typing. A total of400 subjects (33.3%) were nasally colonized with S. aureus, and 17 (1.4%) were nasal carriers of MRSA. The analysis did not identify age, gender, and the two subpopulations as predictors for MRSA colonization. MRSA were more likely to harbor the tst gene (p < 0.05). This work highlighted a low prevalence of nasal MRSA carriage, with 52.94% belonging to sequence type (ST) ST22. The remaining isolates were distributed as singletons (ST8, ST1, and ST398), whereas approximately one-third of MRSA was not identified, including three novel spa-types (t13247, t13248, and t13249). Although we highlighted the current clones present in the Tangier community, they are limited in space and time. Therefore, further studies would be required to obtain a comprehensive picture of the dissemination of MRSA in the community, hospital, and livestock.

Shedding of Methicillin-Resistant Staphylococcus aureus (MRSA) by Hospitalized Patients during Procedures

Background Contaminated environmental surfaces contribute to transmission of healthcare-associated pathogens such as methicillin-resistant Staphylococcus aureus (MRSA). We hypothesized that medical and non-medical procedures facilitate environmental dissemination of MRSA in hospitalized patients.Methods We conducted an observational cohort study of hospitalized MRSA-colonized patients to determine the frequency of and risk factors for environmental shedding during procedures. Prior to each procedure, surfaces in the room and portable equipment used for procedures were disinfected. After procedures, high-touch surfaces and portable equipment were cultured; negative control cultures were collected after 1 hour in the absence of a procedure. Bivariate analyses were performed to identify factors associated with environmental shedding.ResultsOf 55 MRSA colonized patients, 22 (40%) had wounds and 25 (46%) had positive skin cultures. Environmental cultures were collected after 138 total procedures (range, 2 to 12 per patient). As shown in the figure, contamination of surfaces occurred frequently during procedures, but was uncommon in the absence of a procedure. Contamination occurred frequently on surfaces touched by personnel during procedures (12 of 38, 32% positive) and on portable equipment used for procedures (25 of 101, 25%). The presence of a wound was the only factor significantly associated with shedding (59% vs. 26%; P = 0.04).Conclusion Environmental shedding of MRSA occurs frequently during medical and non-medical procedures in hospitalized patients. Our results suggest that there is a need for effective strategies to disinfect surfaces and equipment after procedures.Disclosures All authors: No reported disclosures.

Frequency of Methicillin-Resistant Staphylococcus aureus (MRSA) in Fattening Pigs in the State of Rio Grande do Sul, Brazil

Background: Staphylococcus (S.) aureus is an important nosocomial pathogen in humans and animals worldwide. The commonest class of antibiotics used to treat staphylococcal infections is the β-lactams. Frequently, S. aureus strains show high resistance to methicillin and other β-lactam antibiotics, called “Methicillin-resistant Staphylococcus aureus” (MRSA). Although MRSA has emerged at slower rate in domestic animals, it has frequently been found in the nasal cavity of healthy piglets and its transmission between pigs and swine handlers has already been studied. The aim of this work was to assess the presence of MRSA in finishing pigs in the state of Rio Grande do Sul, Brazil.Materials, Methods &amp; Results: A total of 350 nasal swabs were collected from 10 to 20 week old finishing pigs. Sampling was performed in five pig farms in northeast Rio Grande do Sul State, Brazil. Swabs were stored in tubes without transport medium and carried to the laboratory under refrigeration. The specimens were cultured in selective and differential Agar (Baird Parker) and then were incubated at 37ºC for 48 h. After isolation of typical colonies of S. aureus, they were inoculated in BHI (Brain Heart Infusion) broth at 37ºC for 24 h and tested for tube coagulase activity. Coagulase positive samples were selected for growth in Oxacillin Resistant Screening Agar (ORSA) supplemented with 2 mg/L of oxacillin. This media contains aniline blue to demonstrate mannitol fermentation. Oxacillin and 5.5% NaCl have the capacity to reduce the growth of non-staphylococcal bacteria, selecting for MRSA. Blue colonies growth after 24 to 48 h of incubation at 37ºC indicate the presence of positive MRSA strains. Specimens with at least one colony growing in ORSA within 48 h were considered resistant. Linear regression was performed in order to identify the association between herd size and MRSA frequency (SAS 9.4, 2012). Growth of S. aureus occurred on 18.0% of the samples and differences among farms were found. However, after incubation in ORSA only 18 (5.1%) were MRSA positive, ranging from zero to 12.5% among farms. Significant correlation between herd size and MRSA frequency (adjusted r² = 0.978; P = 0.001) was observed.Discussion: In a previous study in pig herds in Brazil examining swine nasal swabs, 22.5% was positive for S. aureus and none for MRSA. One of 5 farms tested in our work also had no positive animals and 4 of them showed low frequencies, ranging from 1.7% to 12.5% with an average of 5.1%. Our results were similar to those found in Asian countries, but were very different from European data. Some factors can be associated with MRSA frequency in pig farms, such as dust, air contamination, poor hygiene, age, herd size, replacement rate and number of sources. In the present work we found a strong positive correlation (r² adjusted = 0.978; P = 0.001) between herd size and MRSA frequency, such as detected by previous authors. It might occur due to a higher risk of bacterial introduction and higher pressure of infection, easing dissemination of MRSA. Herd size may be a crucial factor to explain the frequencies found, since farms had very similar facilities and handling practices. Although the average frequency has been low in this work when compared to other countries, MRSA was present in almost all farms. This bacteria is able to transmit mecA gene to S. aureus susceptible populations, increasing MRSA frequencies over time.

MRSAspat1081, a Highly Transmissible Strain Endemic to Hong Kong, China, in the Netherlands

MRSA<i>spa</i>t1081, a Highly Transmissible Strain Endemic to Hong Kong, China, in the Netherlands

Spreading of β-lactam resistance gene (mecA) and methicillin-resistant Staphylococcus aureus through municipal and swine slaughterhouse wastewaters

Methicillin-resistant Staphylococcus aureus (MRSA) is a potential zoonotic agent. Municipal wastewater treatment plants (WWTPs) can be reservoirs for MRSA dissemination. It is unclear, however, whether MRSA and its β-lactam resistance gene (mecA) can be spread from WWTPs that treat the wastewater of swine auction markets. The aims of the study were to compare (1) the abundance of the mecA gene in one municipal (M-) and one swine (S-) WWTP and (2) the genotypic and phenotypic characteristics of MRSA isolates from these two types of WWTPs. The concentrations of mecA gene from 96 wastewater samples were quantified using real-time quantitative polymerase chain reaction (real-time qPCR). One hundred and thirteen MRSA isolates were recovered and were characterized by antimicrobial susceptibility testing, minimum inhibitory concentrations (MICs), and staphylococcal cassette chromosome mec (SCCmec) typing. The mecA gene could be detected in all the wastewater samples. A high abundance of recovered mecA gene (2.6 × 101 to 1.9 × 104 gene copies μg−1 of total DNA) in swine slaughterhouse wastewater implied a correspondingly high transferring/receiving potential. All MRSA isolates were multidrug resistant (MDR) and showed high MICs to different antimicrobials. The M-WWTP MRSA isolates harbored SCCmec II–IV and VII, whereas those from the S-WWTP harbored SCCmec V and IX. In conclusion, wastewater from swine slaughterhouses can make these slaughterhouses potential hotspots for the dissemination of mecA gene and MRSA, and the high MICs of MRSA from both WWTP origins may pose a health risk not only to workers but also to the general public.

Transferable Vancomycin Resistance in a Community-Associated MRSA Lineage

We report the case of a patient from Brazil with a bloodstream infection caused by a strain of methicillin-resistant Staphylococcus aureus (MRSA) that was susceptible to vancomycin (designated BR-VSSA) but that acquired the vanA gene cluster during antibiotic therapy and became resistant to vancomycin (designated BR-VRSA). Both strains belong to the sequence type (ST) 8 community-associated genetic lineage that carries the staphylococcal chromosomal cassette mec (SCCmec) type IVa and the S. aureus protein A gene (spa) type t292 and are phylogenetically related to MRSA lineage USA300. A conjugative plasmid of 55,706 bp (pBRZ01) carrying the vanA cluster was identified and readily transferred to other staphylococci. The pBRZ01 plasmid harbors DNA sequences that are typical of the plasmid-associated replication genes rep24 or rep21 described in community-associated MRSA strains from Australia (pWBG745). The presence and dissemination of community-associated MRSA containing vanA could become a serious public health concern.

Massive Dissemination of Methicillin ResistantStaphylococcus aureusin Bloodstream Infections in a High MRSA Prevalence Country: Establishment and Diversification of EMRSA-15

Portugal is the European country with the highest prevalence of methicillin resistant Staphylococcus aureus (MRSA), in which EMRSA-15 (ST22-IVh) has been the dominant clone since soon after its introduction in Portuguese hospitals in 2001. In this study, we intend to not only, assess the evolution of the invasive MRSA in Portuguese hospitals, but also to evaluate the invasive methicillin susceptible S. aureus (MSSA) population and the relationship between both populations. In the current study, two major MRSA clones were identified: EMRSA-15 that has been dominant for more than 10 years and accounts for 75% of the MRSA isolates, and ST105-II, a clone related with the New York/Japan clone (ST5-II). In contrast, among MSSA, several clonal backgrounds were identified. Despite of the massive predominance of EMRSA-15 in the last decade, an increase in spa diversity has been observed in the last few years, which suggests a recent and local diversification of this clone. Interestingly, MRSA and MSSA populations with related clonal backgrounds appear to have increased as a result of the dissemination of MRSA to the community environment.

Current status of Staphylococcus aureus infection in a central teaching hospital in Shanghai, China

BackgroundTo control the spread of methicillin-resistant Staphylococcus aureus (MRSA) in hospitals, infection control measures such as hand hygiene practices were introduced into the teaching hospitals in Shanghai, China, in 2008. Currently, there is limited information characterizing the latest hospital-acquired S. aureus infections in this area. Therefore, we sought to determine the prevalence, molecular characteristics, and genotype-phenotype correlation of hospital-acquired S. aureus infections in Huashan Hospital, one of the largest teaching hospitals in Shanghai.ResultsAmong 608 hospital-acquired S. aureus clinical isolates obtained from January to December of 2011 in Huashan Hospital, 68.1% were MRSA. The predominant MRSA clones were ST239-SCCmecIII and ST5-SCCmecII. ST239 was mainly recovered from respiratory specimens and sterile body fluids, ST5 was associated with respiratory specimens and blood, and ST1 was most prevalent in urine samples. In this study, 31 dispersed sequence types (STs) of methicillin-susceptible S. aureus (MSSA) were identified, most of which caused skin/soft tissue infection and bacteremia. The frequencies of pvl-, muPA-, and qacA/B-positive isolates were 1.6, 9.9, and 11.8% respectively. MuPA was more frequently identified in ST1 and ST5, and qacA/B was more prevalent in ST239 and ST5. Most of the pvl-positive isolates were MSSA, whereas the majority of muPA- and qacA/B-positive isolates were MRSA. ST239 and ST5 had higher resistance rates to multiple antibiotics. In Huashan Hospital, the infection rate in the intensive care unit (ICU) was 3.9 per 1000 hospitalized days, but only 1.2 per 1000 hospitalized days in the other wards. Each ward harbored its own dominant STs. Pulsed-field gel electrophoresis showed diversity within the same epidemic S. aureus clones originating from the same wards.ConclusionThere is still a high prevalence of MRSA infections in the teaching hospital in Shanghai. There were also differences in the major infection types caused by MRSA and MSSA, and hospital-acquired S. aureus infections in the ICU of Huashan Hospital pose a greater threat to patient safety than in other wards. The high proportion of multiple antibiotic and chlorhexidine-based antiseptic-resistant clones in this hospital underscores the need for more effective infection control measures to help curtail dissemination of MRSA to hospitalized patients.

Evidence of possible methicillin-resistant Staphylococcus aureus ST398 spread between pigs and other animals and people residing on the same farm

Methicillin-resistant Staphylococcus aureus (MRSA) has emerged in a wide variety of animal species. However, little is known about the transmission routes of MRSA ST398 between different animal species, the barn environment and people residing on the same farm. In this study, two pig farms, two poultry-pig and two dairy-pig farms were investigated with respect to the presence of MRSA. On each farm, samples were collected from all animal species present, the barn environment, the farmer, household members and the herd veterinarians. Besides the MRSA prevalence, the obtained spa-, SCCmec-type and antimicrobial susceptibility profiles were also compared. Multilocus sequence typing (MLST) showed that MRSA ST398 was found in all animal species, in humans present on the farms and also in the pig barn environment. The presence of MRSA with the same spa-, SCCmec-type and antibiotic profile in the different animal species in direct or indirect contact with pigs suggests MRSA transfer. Furthermore, different pig age categories were investigated, with weaned piglets having the highest MRSA prevalence (86.3%). The herd-level prevalence was highly correlated (r=0.86, p=0.03) between sows and pre-weaned piglets. The results also indicate that companion animals, rats, mice and farmers could play an important role in the dissemination of MRSA, emphasizing the importance of internal biosecurity. However, external biosecurity is equally important because other spa-, SCCmec-types or antimicrobial resistances can be introduced through purchase of gilts. In this study we demonstrated that MRSA likely spreads between animal species, humans and the pig barn environment, which is why it is important to accurately implement control practices, in which not only pigs should be targeted, but also all other animal species present on farms.