Detection of Vancomycin Resistance among Methicillin-Resistant Staphylococcus aureus Strains Recovered from Children with Invasive Diseases in a Reference Pediatric Hospital.

Vancomycin is the cornerstone in treating methicillin-resistant Staphylococcus aureus (MRSA) infections. However, therapeutic failures can occur when MRSA strains with decreased susceptibility to glycopeptides (DSG) are involved. The aim of this study was to detect and characterize DSG in MRSA recovered from children with invasive diseases at a reference pediatric hospital between 2009 and 2019. Fifty-two MRSA strains were screened using agar plates with vancomycin 3 and 4 mg/L (BHI-3 and BHI-4); the VITEK2 system; and standard and macro E-tests. Suspicious hVISA were studied by population analysis profiling-area under the curve (PAP-AUC), and wall thickness was analyzed by transmission electron microscopy. Neither VRSA nor VISA were detected in this set. As only three strains met the hVISA criteria, the PAP-AUC study included 12 additional MRSA strains that grew one colony on BHI-4 plates or showed minimum inhibitory concentrations of vancomycin and/or teicoplanin ≥ 1.5 mg/L. One strain was confirmed as hVISA by PAP-AUC. The wall thickness was greater than the vancomycin-susceptible control strain; it belonged to ST30 and carried SCCmec IV. As expected, a low frequency of hVISA was found (1.9%). The only hVISA confirmed by PAP-AUC was not detected by the screening methods, highlighting the challenge that its detection represents for microbiology laboratories.

Risk factors and outcomes associated with vancomycin-resistant Enterococcus faecium and ampicillin-resistant Enterococcus faecalis bacteraemia: A 10-year study in a tertiary-care centre in Mexico City.

OBJECTIVES: We sought to identify risk factors associated with vancomycin-resistant Enterococcus faecium (VRE) and ampicillin-resistant Enterococcus faecalis (ARE) bacteraemia, predictors of 30-day mortality, and 90-day recurrence-free survival according to resistance. METHODS: We evaluated clinical records of patients with E. faecalis and E. faecium bacteraemia (2007-2017). We performed bivariate and multivariate logistic regression analyses to identify factors associated with VRE and ARE bacteraemia and predictors of 30-day mortality. A Kaplan-Meier estimate of 90-day recurrence-free survival was done. RESULTS: We identified 192 and 147 E. faecium and E. faecalis bacteraemia episodes, respectively, of which 55.7% of E. faecium were VRE (94% vanA) and 12.2% of E. faecalis were ARE. Factors related to VRE bacteraemia were previous hospitalisation (aOR, 80.18, 95% CI 1.81-634), history of central venous catheter (aOR, 11.15, 95% CI 2.48-50.2) and endotracheal cannula use (aOR, 17.91, 95% CI 1.22-262.82). There was higher attributable mortality to VRE (28%, 95% CI 14-68%; P < 0.001) and ARE (10%, 95% CI 0.1-36%; P = 0.58) compared with their susceptible counterparts. APACHE II (aOR, 1.45, 95% CI 1.26-1.66) and history of chemotherapy (aOR, 3.52, 95% CI 1.09-11.39) were predictors of E. faecium bacteraemia 30-day mortality. We could not recognise any factor related to ARE bacteraemia or E. faecalis 30-day mortality. CONCLUSION: History of hospitalisation and invasive device use were related to VRE bacteraemia. APACHE II and history of chemotherapy were predictors of mortality. We could not identify factors related to ARE or predictors of mortality.

Trend of clinical vancomycin-resistant enterococci isolated in a regional Italian hospital from 2001 to 2018.

A retrospective study of the epidemiology of vancomycin-resistant enterococci (VRE) in a regional hospital of central Italy in 2001-2018 demonstrated an increased VRE prevalence since 2016. A total of 113 VRE isolates, 89 E. faecium (VREfm) and 24 E. faecalis (VREfs), were collected in the study period. All strains showed high-level resistance to vancomycin; 107 also showed teicoplanin resistance. Altogether, 84 VREfm and 20 VREfs carried vanA, whereas 5 VREfm and 1 VREfs carried vanB. MLST analysis documented that 89 VREfm isolates mainly belonged to ST78, ST80, and ST117. Most strains were isolated from 2001 to 2007, ST78 being the predominant clone. VREfm re-emerged in 2016 with a prevalence of the ST80 lineage. Most VREfs were isolated from 2001 to 2006; although they belonged to 7 different STs, there was a prevalence of ST88 and ST6. Notably, ST88 was sporadically recovered throughout the study period. The increasing rate of VREfm isolation from 2016 to 2018 may be related to the influx of new successful clones and to the renewed and widespread use of vancomycin. Improved infection control measures in hospital wards should be adopted to limit the spread of new epidemic VRE strains.

High Phenotypic and Genotypic Diversity of Enterococcus faecium from Clinical and Commensal Isolates in Third Level Hospital.

Background: The use of antimicrobials and myeloablative chemotherapy regimens has promoted multiresistant microorganisms to emerge as nosocomial pathogens, such as vancomycin-resistant Enterococcus faecium (VREfm). We described a polyclonal outbreak of bloodstream infection caused by Efm in a hemato-oncological ward in Mexico. Our aim was to describe the clonal complex (CC) of the Efm strains isolated in the outbreak in comparison with commensal and environmental isolates. Methodology: Sixty Efm clinical, environmental, and commensal strains were included. We constructed a cladogram and a phylogenetic tree using Vitek and Multilocus sequence typing data, respectively. Results: We reported 20 new sequence types (ST), among which 17/43 clinical isolates belonged to CC17. The predominant ST in the clinical strains were ST757, ST1304, ST412, and ST770. Neither environmental nor commensal isolates belonged to CC17. The phylogeny of our collection shows that the majority of the clinical isolates were different from the environmental and commensal isolates, and only a small group of clinical isolates was closely related with environmental and commensal isolates. The cladogram revealed a similar segregation to that of the phylogeny. Conclusions: We found a high diversity among clinical, environmental, and commensal strains in a group of samples in a single hospital. Highest diversity was found between commensal and environmental isolates.

Assessment of vancomycin resistance transfer among enterococci of clinical importance in milk matrix.

Dissemination of vancomycin resistance in enterococci has been associated with horizontal transfer of mobile genetic elements. Aim of the study was to evaluate if milk matrix is a suitable environment to support transferability of vancomycin resistance (vanA) gene from clinical vancomycin-resistant Enterococcus faecium to vancomycin-sensitive Enterococcus faecalis. Enterococci strains were firstly screened for the presence of cpd (inducible sex pheromone determinant) gene, vanA and tetL genes (vancomycin and tetracycline resistance markers, respectively) and the gelE (extracellular metalloendopeptidase) gene to define the mating pairs. Based on these selection markers, we investigated the transferability of eight plasmid-borne vanA harbored by E. faecium (vanA+, cpd-, tetL- and gelE-) into two E. faecalis (vanA-, cpd+, tetL + and gelE+) recipient strains in milk matrix. The strains were mated in a 1:1 ratio in 7% reconstituted milk and incubated at 37 °C. Transconjugants emerged from all 16 matings within 2 h of incubation and were evidenced by dual antibiotic resistance (vancomycin and tetracycline). The vancomycin-resistance of trasconjugants was maintained even after ten subsequent passages on nonselective medium. Transconjugants were positive for vanA, tetL and gelE genes. This study indicates milk matrix as suitable environment to support gene exchange between Enterococcus species.

Colonización por enterococo resistente a vancomicina en pacientes internados en un hospital de Lima, Perú / Colonization by Enterococcal Strains Resistant to Vancomycin in Patients from a Hospital in Lima, Peru

RESUMEN Con el objetivo de determinar la frecuencia de colonización por el enterococo resistente a vancomicina (ERV), el genotipo de resistencia y los factores asociados, se realizó un estudio de tipo transversal durante noviembre y diciembre del 2013 en el Hospital Nacional Cayetano Heredia en Lima, Perú. Se encontró una frecuencia de colonización por ERV de 6,2% (IC 95%: 1,67-10,73), todas las cepas aisladas tenían el genotipo de resistencia vanA, y se halló que las variables hospitalización previa (p=0,001) y el uso de cefalosporinas de tercera generación (p=0,016) estaban asociadas a la colonización por ERV. En conclusión, existe colonización perianal por ERV en los diversos servicios de hospitalización, el gen vanA podría ser transmitido a gérmenes más virulentos y ocasionar la aparición de la bacteria Staphylococcus aureus resistente a vancomicina (VRSA). Es necesario adoptar medidas de control de infecciones para evitar la transmisión de esta bacteria en el ambiente hospitalario.

ABSTRACT This cross-sectional study was conducted from November to December of 2013 at the Cayetano Heredia National Hospital in Lima, Peru, to determine the rate of infection with vancomycin-resistant enterococcus (VRE), the resistance genotype, and associated factors. The rate of infection with VRE was 6.2% (95% confidence interval [CI]: 1.67-10.73) and the resistance genotype isolated from all strains was the vanA gene. The factors associated with colonization with VRE were previous hospitalizations (p = 0.001) and the use of third-generation cephalosporins (p = 0.016). In conclusion, perianal colonization with VRE is present in many hospital services. Moreover, the vanA gene may cause resistance to vancomycin and promote the development of vancomycin-resistant Staphylococcus aureus. Therefore, infection control measures should be adopted to prevent the dissemination of this bacterial strain in hospital settings.

Detection of vancomycin-resistant enterococci (VRE) in stool specimens submitted for Clostridium difficile toxin testing

The aim of this study was to determine the association between Clostridium difficile (C. difficile) and vancomycin-resistant Enterococcus (VRE) and efficacy of screening stools submitted for C. difficile toxin assay for prevalence of VRE. Between April 2012 and February 2014, 158 stool samples submitted for C. difficile toxin to the Marmara University Microbiology Laboratory, were included in the study. Stool samples were analyzed by enzyme immuno assay test; VIDAS (bioMerieux, France) for Toxin A&B. Samples were inoculated on chromID VRE (bioMerieux, France) and incubated 24 h at 37 °C. Manuel tests and API20 STREP (bioMerieux, France) test were used to identify the Enterococci species. After the species identification, vancomycin and teicoplanin MIC's were performed by E test and molecular resistance genes for vanA vs vanB were detected by polymerase chain reaction (PCR). Of the 158 stool samples, 88 were toxin positive. The prevalence of VRE was 17%(n:19) in toxin positives however, 11.4% in toxin negatives(n:70). All VRE isolates were identified as Enterococcus faecium. These results were evaluated according to Fischer's exact chi-square test and p value between VRE colonization and C. difficile toxin positivity was detected 0.047 (p 0.05). PPV and NPV were 79% and 47% respectively. In our study, the presence of VRE in C. difficile toxin positives is statistically significant compared with toxin negatives (p 0.05). Screening for VRE is both additional cost and work load for the laboratories. Therefore VRE screening among C. difficile toxin positive samples, will be cost effective for determination of high risk patients in the hospitals especially for developing countries.(AU)

Detection of vancomycin-resistant enterococci (VRE) in stool specimens submitted for Clostridium difficile toxin testing

Abstract The aim of this study was to determine the association between Clostridium difficile (C. difficile) and vancomycin-resistant Enterococcus (VRE) and efficacy of screening stools submitted for C. difficile toxin assay for prevalence of VRE. Between April 2012 and February 2014, 158 stool samples submitted for C. difficile toxin to the Marmara University Microbiology Laboratory, were included in the study. Stool samples were analyzed by enzyme immuno assay test; VIDAS (bioMerieux, France) for Toxin A&B. Samples were inoculated on chromID VRE (bioMerieux, France) and incubated 24 h at 37 °C. Manuel tests and API20 STREP (bioMerieux, France) test were used to identify the Enterococci species. After the species identification, vancomycin and teicoplanin MIC's were performed by E test and molecular resistance genes for vanA vs vanB were detected by polymerase chain reaction (PCR). Of the 158 stool samples, 88 were toxin positive. The prevalence of VRE was 17%(n:19) in toxin positives however, 11.4% in toxin negatives(n:70). All VRE isolates were identified as Enterococcus faecium. These results were evaluated according to Fischer's exact chi-square test and p value between VRE colonization and C. difficile toxin positivity was detected 0.047 (p < 0.05). PPV and NPV were 79% and 47% respectively. In our study, the presence of VRE in C. difficile toxin positives is statistically significant compared with toxin negatives (p < 0.05). Screening for VRE is both additional cost and work load for the laboratories. Therefore VRE screening among C. difficile toxin positive samples, will be cost effective for determination of high risk patients in the hospitals especially for developing countries.

Detection of vancomycin-resistant enterococci (VRE) in stool specimens submitted for Clostridium difficile toxin testing.

The aim of this study was to determine the association between Clostridium difficile (C. difficile) and vancomycin-resistant Enterococcus (VRE) and efficacy of screening stools submitted for C. difficile toxin assay for prevalence of VRE. Between April 2012 and February 2014, 158 stool samples submitted for C. difficile toxin to the Marmara University Microbiology Laboratory, were included in the study. Stool samples were analyzed by enzyme immuno assay test; VIDAS (bioMerieux, France) for Toxin A&B. Samples were inoculated on chromID VRE (bioMerieux, France) and incubated 24h at 37°C. Manuel tests and API20 STREP (bioMerieux, France) test were used to identify the Enterococci species. After the species identification, vancomycin and teicoplanin MIC's were performed by E test and molecular resistance genes for vanA vs vanB were detected by polymerase chain reaction (PCR). Of the 158 stool samples, 88 were toxin positive. The prevalence of VRE was 17%(n:19) in toxin positives however, 11.4% in toxin negatives(n:70). All VRE isolates were identified as Enterococcus faecium. These results were evaluated according to Fischer's exact chi-square test and p value between VRE colonization and C. difficile toxin positivity was detected 0.047 (p<0.05). PPV and NPV were 79% and 47% respectively. In our study, the presence of VRE in C. difficile toxin positives is statistically significant compared with toxin negatives (p<0.05). Screening for VRE is both additional cost and work load for the laboratories. Therefore VRE screening among C. difficile toxin positive samples, will be cost effective for determination of high risk patients in the hospitals especially for developing countries.

[Methicillin resistance and vancomycin susceptibility pattern among blood isolates of Staphylococcus aureus]. / Resistencia a meticilina y susceptibilidad a vancomicina de Staphylococcus aureus aislados de sangre.

BACKGROUND: Staphylococcus aureus is capable of acquiring resistance against all antimicrobial agents. Vancomycin has been the cornerstone therapy for serious methicillin-resistant S. aureus infections. However, vancomycin treatment failures have been reported. METHODS: From March to August 2010, S. aureus blood isolates were included to determine methicillin-resistance and the vancomycin susceptibility by using a standard microdilution method. To detect methicillin-resistance, a Mueller-Hinton agar plate added with oxacillin 4 µg/mL and 2 % NaCl, and an agglutination test were used. Growth of S. aureus on the agar plate and/or reactive agglutination defined a methicillin-resistant organism. Vancomycin susceptibility was assessed by determining the minimal inhibitory concentration (MIC) in Muelller-Hinton agar plates prepared with dilutions ranging from 16 to 0.5 µg/mL. RESULTS: A total of 25 blood-isolates of S. aureus were included. A 60 % was methicillin-resistant. All isolates were vancomycin-susceptible (MIC ≤ 2 µg/mL) showing the next MICs distribution: 48 % ≤ 0.5 µg/mL; 44 % 1 µg/mL, and 8 % 2 µg/mL. CONCLUSION: The high proportion of methicillin-resistance among S. aureus and the presence of vancomycin susceptible phenotypes (MIC of 2 µg/mL) not only claim for an enforcement of standard precautions and antimicrobial control, but also for a regular surveillance of vancomycin susceptibility pattern using a reference method.

Introducción: el Staphylococcus aureus es capaz de desarrollar resistencia a todos los antimicrobianos. La vancomicina es clave para tratar infecciones graves causadas por S. aureus meticilino-resistente. Sin embargo, últimamente se reportan fallas terapéuticas. El objetivo fue establecer la resistencia a la meticilina y el perfil de susceptibilidad a la vancomicina del S. aureus. Métodos: de marzo a agosto del 2010, se determinó la meticilino-resistencia y la susceptibilidad a vancomicina de S. aureus aislados de hemocultivos, mediante el método estándar de microdilución. Para la meticilino-resistencia se utilizó una placa de agar Mueller-Hinton con 4 µg/mL de oxacilina, más NaCl al 2 % y una prueba de aglutinación. El desarrollo bacteriano o la aglutinación positiva identificaron al microorganismo meticilino-resistente. Para la susceptibilidad a vancomicina se determinó la concentración mínima inhibitoria (CMI) en placas de agar Mueller-Hinton con dilución de 16 a 0.5 µg/mL. Resultados: en total se incluyeron 25 S. aureus. El 60 % fue meticilino-resistente; el 100 % sensible a vancomicina (CMI ≤ 2 µg/mL), con las siguientes CMI: el 48 %, ≤ 0.5 µg/mL; 44 %, 1 µg/mL; y el 8 %, 2 µg/mL. Conclusión: la proporción alta de meticilino-resistencia y la evidencia de fenotipos sensibles a la vancomicina, pero asociados a falla terapéutica (CMI 2 µg/mL), demandan no solo el reforzamiento continuo de las precauciones estándar y el control de antimicrobianos sino también la vigilancia sistemática del patrón de susceptibilidad a la vancomicina con un método de referencia.

Phenotypic and genotypic characterization of vancomycin-resistant Enterococcus faecium clinical isolates from two hospitals in Mexico: First detection of VanB phenotype-vanA genotype.

INTRODUCTION: Enterococcus faecium has emerged as a multidrug-resistant nosocomial pathogen involved in outbreaks worldwide. Our aim was to determine the antimicrobial susceptibility, biofilm production, and clonal relatedness of vancomycin-resistant E. faecium (VREF) clinical isolates from two hospitals in Mexico. METHODS: Consecutive clinical isolates (n=56) were collected in two tertiary care hospitals in Mexico from 2011 to 2014. VREF isolates were characterized by phenotypic and molecular methods including pulsed-field gel electrophoresis (PFGE). RESULTS: VREF isolates were highly resistant to vancomycin, erythromycin, norfloxacin, high-level streptomycin, and teicoplanin, and showed lower resistance to tetracycline, nitrofurantoin and quinupristin-dalfopristin. None of the isolates were resistant to linezolid. The vanA gene was detected in all isolates. Two VanB phenotype-vanA genotype isolates, highly resistant to vancomycin and susceptible to teicoplanin, were detected. Furthermore, 17.9% of the isolates were classified as biofilm producers, and the espfm gene was found in 98.2% of the isolates. A total of 37 distinct PFGE patterns and 6 clones (25% of the isolates as clone A, 5.4% as clone B, and 3.6% each as clone C, D, E, and F) were detected. Clone A was detected in 5 different wards of the same hospital during 14 months of surveillance. CONCLUSION: The high resistance to most antimicrobial agents and the moderate cross-transmission of VREF detected accentuates the need for continuous surveillance of E. faecium in the hospital setting. This is also the first reported incidence of the E. faecium VanB phenotype-vanA genotype in the Americas.

Methicillin-Susceptible, Vancomycin-Resistant Staphylococcus aureus, Brazil.

We report characterization of a methicillin-susceptible, vancomycin-resistant bloodstream isolate of Staphylococcus aureus recovered from a patient in Brazil. Emergence of vancomycin resistance in methicillin-susceptible S. aureus would indicate that this resistance trait might be poised to disseminate more rapidly among S. aureus and represents a major public health threat.

Tendencias de la resistencia a antibióticos en Medellín y en los municipios del área metropolitana entre 2007 y 2012: resultados de seis años de vigilancia / Trends in antibiotic resistance in Medellín and municipalities of the Metropolitan Area between 2007 and 2012: Results of six years of surveillance

Introducción. La resistencia bacteriana es un fenómeno mundial, pero su comportamiento varía en el tiempo y el espacio, confiriéndole importancia a los sistemas de vigilancia locales. Objetivo. Determinar las tendencias de la resistencia a antibióticos entre 2007 y 2012 en instituciones hospitalarias de Medellín y del Área Metropolitana del Valle de Aburrá. Materiales y métodos. Entre 2007 y 2012 se obtuvieron los porcentajes de resistencia a antibióticos marcadores en 22 instituciones, utilizando el programa Whonet 5.6. Se empleó la guía del Clinical and Laboratory Standards Institute (CLSI) de los años 2009 y 2012 para interpretar los resultados de las pruebas de sensibilidad. Con el programa Epi-Info 6.04 se analizaron tendencias por medio de la prueba de ji al cuadrado de tendencia lineal con un nivel de confianza de 95 %; se consideró significativo un valor de p=0,05. Resultados. Se observó una disminución de la resistencia a oxacilina en S taphylococcus aureus (p=0,0006) y un incremento de la resistencia a vancomicina en Enterococcus faecium (p=0,0000). En Escherichia coli y Serratia marcescens se observó un incremento de la resistencia a ceftazidima, en contraste con una disminución en Klebsiella pneumoniae (p=0,0000) y Enterobacter cloacae (p=0,058). Para K. pneumoniae, S. marcescens y E. cloacae se observó un incremento de la resistencia a carbapenémicos, en contraste con una disminución en Pseudomonas aeruginosa y Acinetobacter baumannii. Conclusiones. La vigilancia de la resistencia permitió obtener hallazgos importantes como la emergencia de E. faecium resistente a la vancomicina y enterobacterias resistentes a los carbapenémicos. Es indispensable conocer el uso de antibióticos en la región para establecer su influencia en los perfiles encontrados, además de garantizar la calidad de la información emanada de los laboratorios de microbiología.

Introduction: Bacterial resistance is a global phenomenon, but it presents geographic and temporal variations; this is the importance of local surveillance programs. Objective: To determine trends in antibiotic resistance in hospitals between 2007 and 2012 in Medellín and its Metropolitan Area. Materials and methods: Percentages of antibiotic resistance between 2007 and 2012 in 22 institutions were obtained using WHONET 5.6 program. For interpretation of susceptibility results, CLSI standards of 2009 and 2012 were used. Using the Epi-Info 6.04 program a trends analysis of antibiotic resistance was done using the chi-square for linear trend with a confidence level of 95%, a value of p=0.05 was considered significant. Results: In six years of surveillance of antibiotic resistance we found a decrease of oxacillin resistance in Staphylococcus aureus (p=0.0006) and an increase of vancomycin resistance in Enterococcus faecium (p=0.0000). In Escherichia coli and Serratia marcescens an increase of resistance to ceftazidime was found, in contrast to a decrease in Klebsiella pneumoniae (p=0.0000) and Enterobacter cloacae (p=0.058). K. pneumoniae , S. marcescens and E. cloacae showed an increase of carbapenem resistance in contrast to a reduction of carbapenem resistance in Pseudomonas aeruginosa and Acinetobacter baumannii . Conclusions: The resistance surveillance identified important findings as the emergence of E. faecium resistant to vancomycin and carbapenem-resistant Enterobacteriaceae . It is essential to determine the antibiotic use in the region to establish their influence on the resistance profiles, as well as ensuring the quality of information and microbiological procedures in the microbiology laboratories.

Presence of virulence factors in Enterococcus faecalis and Enterococcus faecium susceptible and resistant to vancomycin

Despite the increasing importance of Enterococcus as opportunistic pathogens, their virulence factors are still poorly understood. This study determines the frequency of virulence factors in clinical and commensal Enterococcus isolates from inpatients in Porto Alegre, Brazil. Fifty Enterococcus isolates were analysed and the presence of the gelE, asa1 and esp genes was determined. Gelatinase activity and biofilm formation were also tested. The clonal relationships among the isolates were evaluated using pulsed-field gel electrophoresis. The asa1, gelE and esp genes were identified in 38%, 60% and 76% of all isolates, respectively. The first two genes were more prevalent in Enterococcus faecalis than in Enterococcus faecium, as was biofilm formation, which was associated with gelE and asa1 genes, but not with the esp gene. The presence of gelE and the activity of gelatinase were not fully concordant. No relationship was observed among any virulence factors and specific subclones of E. faecalis or E. faecium resistant to vancomycin. In conclusion, E. faecalis and E. faecium isolates showed significantly different patterns of virulence determinants. Neither the source of isolation nor the clonal relationship or vancomycin resistance influenced their distribution.

Detection of vanC 1 gene transcription in vancomycin-susceptible Enterococcus faecalis

Here we report the presence and expression levels of the vanC 1 and vanC 2/3 genes in vancomycin-susceptible strains of Enterococcus faecalis. The vanC 1 and vanC 2/3 genes were located in the plasmid DNA and on the chromosome, respectively. Specific mRNA of the vanC 1 gene was detected in one of these strains. Additionally, analysis of the vanC gene sequences showed that these genes are related to the vanC genes of Enterococcus gallinarum and Enterococcus casseliflavus. The presence of vanC genes is useful for the identification of E. gallinarum and E. casseliflavus. Moreover, this is the first report of vanC mRNA in E. faecalis.

Enterococcus spp.: Resistencia antimicrobiana en infecciones intrahospitalarias / Enterococcus spp.: Antimicrobial resistance in hospital- acquired infections / Enterococcus spp.: ResistÛncia antimicrobiana em infecþ§es intra-hospitalares

Entre los años 1996 y 2010 se estudiaron 1873 aislamientos de Enterococcus spp. pertenecientes a pacientes internados en un hospital universitario de la Ciudad Autónoma de Buenos Aires con infección intrahospitalaria. El 64,2% y el 30,4% de los aislamientos correspondieron a E. faecalis y E. faecium, respectivamente. En el periodo estudiado las infecciones por Enterococcus spp. representaron entre el 8% y el 10% del total de las infecciones nosocomiales. La prevalencia de E. faecium aumentó de un 1,5% en el año 1996 a un 4% en 2010. El primer aislamiento de enterococo resistente a vancomicina se detectó en el año 1998 y correspondió a un E. faecium y en el año 2004 se halló en E. faecalis. Actualmente más del 70% de los aislamientos de E. faecium son resistentes a vancomicina, no así en E. faecalis donde la resistencia es ocasional. No se detectó resistencia a linezolid ni a tigeciclina en Enterococcus spp.(AU)

One thousand eight hundred and seventy-three Enterococcus spp. isolates belonging to patients undergoing hospital-acquired infections at the University Hospital of Buenos Aires city were studied between the years 1996 and 2010. A total of 64.2% and 30.4% of the isolates were identified as E. faecalis y E. faecium respectively. The infections caused by Enterococcus spp. represented from 8% to 10% of the total number of the nosocomial infections in the period studied. The E. faecium prevalence increased from 1.5% in 1996 to 4% in 2010. The first vancomycin-resistant Enterococcus faecium was detected in the year 1998 and in the year 2004 this resistance was detected in E. faecalis as well. At present, more than 70% of the E. faecium isolates show vancomycin resistance; on the contrary, in the case of E. faecalis, this resistance is unusual. Resistance to linezolid or to tigecycline has not been detected in Enterococcus spp. so far.(AU)

Entre os anos 1996 e 2010 estudaram-se 1873 isolamentos de Enterococcus spp. pertencentes a pacientes internados num Hospital Universitário da Cidade Aut¶noma de Buenos Aires com infecþÒo intra-hospitalar. 64,2% e 30,4% dos isolamentos corresponderam a E. faecalis e E. faecium respectivamente. No estudado as infecþ§es por Enterococcus spp. representaram entre 8% e 10% do total das infecþ§es nosocomiais. A prevalÛncia de E. faecium aumentou de 1,5% no ano 1996 para 4% em 2010. O primeiro isolamento de enterococo resistente a vancomicina foi detectado no ano 1998 e correspondeu a um E. faecium e no ano 2004 foi achado em E. faecalis. Atualmente mais de 70% dos isolamentos de E. faecium sÒo resistentes a vancomicina, mas nÒo é assim em E. faecalis onde a resistÛncia é ocasional. NÒo se detectou resistÛncia a linezolid nem a tigeciclina em Enterococcus spp.(AU)

Successful prevention of the transmission of vancomycin-resistant enterococci in a Brazilian public teaching hospital / Sucesso no controle da transmissão hospitalar de Enterococos resistentes a vancomicina em um hospital universitário público brasileiro

INTRODUCTION: Vancomycin-resistant enterococci (VRE) can colonize or cause infections in high-risk patients and contaminate the environment. Our objective was to describe theepidemiological investigation of an outbreak of VRE, the interventions made, and their impact on its control. METHODS: We conducted a retrospective, descriptive, non-comparative study by reviewing the charts of patients with a VRE-positive culture in the University Hospital of Campinas State University, comprising 380 beds, 40 of which were in intensive care units (ICUs), who were admitted from February 2008-January 2009. Interventions were divided into educational activity, reviewing the workflow processes, engineering measures, and administrative procedures. RESULTS: There were 150 patients, 139 (92.7%) colonized and 11 (7.3%) infected. Seventy-three percent were cared for in non-ICUs (p = 0.028). Infection was more frequent in patients with a central-line (p = 0.043), mechanical ventilation (p = 0.013), urinary catheter (p = 0.049), or surgical drain (p = 0.049). Vancomycin, metronidazole, ciprofloxacin, and third-generation cephalosporin were previously used by 47 (31.3%), 31 (20.7%), 24 (16%), and 24 (16%) patients, respectively. Death was more frequent in infected (73%) than in colonized (17%) patients (p < 0.001). After the interventions, the attack rate fell from 1.49 to 0.33 (p < 0.001). CONCLUSIONS: Classical risk factors for VRE colonization or infection, e.g., being cared for in an ICU and previous use of vancomycin, were not found in this study. The conjunction of an educational program, strict adhesion to contact precautions, and reinforcement of environmental cleaning were able to prevent the dissemination of VRE.

INTRODUÇÃO: Enterococos resistentes a vancomicina (ERV) podem colonizar e causar infecção em pacientes de alto risco, bem como contaminar o ambiente. Nosso objetivo foi descrever a investigação epidemiológica de um surto de ERV, as intervenções realizadas e o impacto no controle do surto. MÉTODOS: Estudo retrospectivo, descritivo, por revisão de prontuários de pacientes com cultura positiva para ERV em um hospital geral, público, universitário, admitidos entre fevereiro de 2008 e janeiro de 2009. As intervenções foram divididas em ações educacionais, revisão de processos de trabalho, medidas administrativas e de engenharia. RESULTADOS: Foram avaliados 150 pacientes, 139 (92,7%) colonizados e 11 (7,3%) infectados por ERV. Setenta e três por cento estavam internados em unidades de cuidados não intensivos (p=0,028). Infecção por ERV foi mais frequente em pacientes usando cateter venoso central (p=0,043), ventilação mecânica (p=0,013), cateter urinário (p=0,049) ou drenos cirúrgicos (p=0,049). Vancomicina, metronidazol, ciprofloxacina ou cefalosporina de terceira geração foram utilizados previamente por 47 (31,3%), 31 (20,7%), 24 (16%) e 24 (16%) pacientes, respectivamente. Óbito foi mais frequente em pacientes infectados por ERV (73%) em relação aos colonizados (17%) (p<0,001). Após as intervenções, a taxa de ataque diminuiu de 1,49 para 0,33 (p<0,001). CONCLUSÕES: Fatores de risco clássicos para colonização ou infecção por ERV, como internação em unidade de terapia intensiva e uso prévio de vancomicina, não foram identificados neste estudo. Um conjunto de intervenções, tais como programa educacional, maior adesão às precauções de contato e reforço da limpeza ambiental apresentou impacto no controle da disseminação hospitalar do ERV.

Resistência antimicrobiana associada em isolados clínicos de Enterococcus spp / Associated antimicrobial resistance in Enterococcus spp. clinical isolates

INTRODUÇÃO: O aumento da prevalência de isolados de enterococos em hospitais, particularmente Enterococcus resistente à vancomicina (VRE), é importante por causa da limitada terapia antimicrobiana efetiva para o tratamento de infecções enterocócicas. MÉTODOS: O presente trabalho apresentou uma investigação retrospectiva de dados de suscetibilidade in vitro quantitativa para uma variedade de antimicrobianos frente aos isolados de Enterococcus spp. e avaliação da associação de resistência entre os agentes antimicrobianos apontados como escolha para o tratamento de infecções causadas por VRE, através do cálculo do risco relativo. RESULTADOS: Dos 156 isolados de enterococos, 40 (25,6 por cento) foram resistentes a três ou mais antimicrobianos, incluindo 7,7 por cento (n = 12/156) resistentes à vancomicina. A associação de resistência elevada foi mais pronunciada entre os isolados de VREs com antimicrobianos alternativos e primários para o tratamento de infecções causadas por estes patógenos, incluindo ampicilina (100 por cento, RR = 7,2), estreptomicina (90,9 por cento, RR = 4,9), rifampicina (91,7 por cento, RR = 3,1) e linezolida (50 por cento, RR = 11,5), apesar da alta taxa de suscetibilidade a esta droga (94,9 por cento). CONCLUSÕES: A resistência associada significativa aos antimicrobianos de primeira escolha e alternativos, usados no tratamento de infecções graves por cepas com o fenótipo VRE e que requerem um regime terapêutico combinado, evidencia alternativas terapêuticas ainda mais limitadas na instituição analisada.

INTRODUCTION: The increasing prevalence of enterococci strains in hospitals, particularly among isolates of vancomycin-resistant enterococci (VRE), poses important problems because of the limited effect of antimicrobial therapy for enterococcal infections. METHODS: This work presents a retrospective investigation of quantitative in vitro susceptibility data for the range of antimicrobials against Enterococcus spp. isolates and evaluation of the association of resistance between antimicrobial agents recommended as the treatment of choice for infections caused by VRE through calculation of the relative risk. RESULTS: Of the 156 enterococci isolates, 40 (25.6 percent) were resistant to 3 or more antimicrobials, including 7.7 percent (n = 12/156) vancomycin resistant. The association of elevated resistance was more pronounced among VRE isolates against alternative and primary antimicrobials for the treatment of infections caused by these pathogens, including ampicillin (100 percent, RR = 7.2), streptomycin (90.9 percent, RR = 4.9), rifampin (91.7 percent, RR = 3.1) and linezolid (50 percent, RR = 11.5), despite high susceptibility to this drug (94.9 percent). CONCLUSIONS: The significant associated resistance to alternative and first choice antimicrobials used in the treatment of serious infections of strains with the VRE phenotype and that require a combined therapeutic regime, revealed even more limited therapeutic alternatives in the institution analyzed.

Community prevalence of methicillin and vancomycin resistant Staphylococcus aureus in and around Bangalore, southern India / Prevalência de Staphylococcus aureus resistente à meticilina e à vancomicina em comunidade no entorno de Bangalore, Índia do Sul

INTRODUCTION: Staphylococcus aureus is a known colonizer in humans and has been implicated in community acquired soft tissue infections. However emergence of methicillin resistant S. aureus (MRSA) has aroused great concern worldwide. This study aimed to determine the prevalence of MRSA in the community of Bangalore, southern India. METHODS: Swabs were collected from anterior nares, forearm, dorsum and palm of the hands of 1,000 healthy individuals residing in and around Bangalore, belonging to different socioeconomic strata and age groups. RESULTS: Analysis verified that 22.5 percent and 16.6 percent of the individuals presented Staphylococcus aureus and MRSA, respectively, at any of the three sites. Vancomycin resistance was observed in 1.4 percent of the S. aureus isolates, which was confirmed by detection of the vanA gene. It was interesting to note that 58.8 percent of the children in the age group 1-5 years-old presented MRSA, the highest percentage compared to other age groups of < 1 (44.4 percent) year-old, 5-20 (21.7 percent) years-old, > 40 (11 percent) years-old and 20-40 (9.9 percent) years-old. Among the population of various socioeconomic strata, maximum MRSA colonization was observed among doctors (22.2 percent), followed by upper economic class (18.8 percent), lower economic class (17.7 percent), apparently healthy hospital in-patients (16.5 percent), nurses (16 percent) and middle economic class (12.5 percent). Most of the MRSA isolates were capsular polysaccharide antigen type 8 (57.1 percent). CONCLUSIONS: There is a need for continuous surveillance and monitoring of the presence of MRSA in the community and a clearer understanding of the dynamics of the spread of MRSA will assist in controlling its dissemination.

INTRODUÇÃO: O Staphylococcus aureus é conhecido por ser um colonizador em humanos sendo implicado em infecções comunitárias dos tecidos moles. Contudo, a resistência à meticilina e emergência de S. aureus meticilina resistentes (MRSA) têm despertado preocupação em todo o mundo. O presente estudo visa encontrar a prevalência de MRSA na comunidade de Bangalore, sul da Índia. MÉTODOS: Suabes foram coletados de narinas anteriores, antebraço e dorso da palma de 1.000 indivíduos saudáveis, residentes em Bangalore e nas proximidades, pertencentes a diferentes estratos socioeconômicos e faixas etárias. RESULTADOS: Observou-se que 22,5 por cento e 16,6 por cento dos indivíduos foram abrigar Staphylococcus aureus e MRSA, respectivamente, em qualquer um dos três locais. Dos S. aureus isolados, 1,4 por cento também foram resistentes à vancomicina, o que foi confirmado pela detecção do gene vanA. Foi interessante notar que 58,8 por cento das crianças na faixa etária de 1-5 anos foram abrigar MRSA, o mais elevado em comparação com outros grupos etários de < 1 (44,4 por cento) ano, 50-20 (21,7 por cento) anos, > 40 (11 por cento) anos e 20-40 (9,9 por cento) anos. Entre a população de diferentes estratos socioeconômicos, a colonização de MRSA máxima foi observada entre os médicos (22,2 por cento), seguida pela classe econômica superior (18,8 por cento), classe baixa (17,7 por cento), pacientes aparentemente saudáveis (16,5 por cento), enfermeiros (16 por cento) e classe econômica média (12,5 por cento). A maioria dos MRSA isolados eram do tipo polissacarídeo capsular antígeno 8 (57,1 por cento). CONCLUSÕES: Há uma necessidade de vigilância e monitorização contínua da presença de MRSA na comunidade, bem como uma melhor compreensão da dinâmica de propagação de MRSA pode ajudar no controle da disseminação.

Absence of intestinal colonization by vancomycin-resistant enterococci in nonhuman primate / Ausência de enterococos resistentes à vancomicina na microbiota intestinal de primatas não-humanos

The animal reservoirs of vancomycin-resistant enterococci (VRE) have important role in the epidemiology of the bacteria and resistant genes. The present work searched fecal samples taken off nonhuman primates for the presence of VRE. Resistance profiles, virulence traits, and genetic variability among enterococci isolates were also analyzed. The samples included Capuchin monkeys (Cebus apella, n=28) and Common marmoset (Callithrix penicillata, n=37) housed in the Primate Center of the University of Brasília, Brazil. Most individuals were captive monkeys from the Central-West and South-East regions of Brazil (n=48). We collected rectal swabs and carried out selective isolation followed by multiplex Polymerase Chain Reaction (PCR) to identify species and resistance genes. No vanA or vanB-containing enterococci were found. The carriage rates ranged from 1.5 percent for the VanC-type E. casseliflavus and E. gallinarum until 12.3 percent (n=8) for Enterococcus faecalis. All E. faecalis isolates showed susceptibility to vancomycin, teicoplanin, ampicillin, gentamicin, and streptomycin. The virulence genes ace and esp were prevalent (100.0 percent, 87.5 percent). Multilocus variable number of tandem repeats (MLVA) revealed diversity in the number of repeats among E. faecalis isolates and targets, which was higher for espC, efa5, and efa6. We identified six different MLVA genotypes that were divergent from those described in human beings. Also, they were clustered into two genogroups that showed host-specificity for the species Cebus apella or Callithrix penicillata. In conclusion, no vanA- or vanB-containing enterococci were found colonizing those primate individuals. This finding suggested that the primate individuals investigated in our study are not directly involved in the epidemiological chain of high-level vancomycin-resistant genes vanA or vanB in Brazil. Our study also showed that E. faecalis isolated from nonhuman primates carry virulence...

Os reservatórios animais de Enterococos Resistentes à Vancomicina (VRE) têm um importante papel na epidemiologia destas bactérias e dos respectivos genes de resistência. O presente estudo examinou a presença de VRE em amostras fecais obtidas de primatas não-humanos. Foram analisados os perfis de resistência, as características de virulência e a variabilidade genética dos isolados. A amostragem incluiu macacos Prego (Cebus apella, n=28) e Sagüis do cerrado (Callithrix penicillata, n=37) alojados no Centro de Primatologia da Universidade de Brasília, Brasil. A maioria dos indivíduos amostrados foram macacos apreendidos na região Centro-Oeste e Sudeste do Brasil (n=48). Assim, foram coletados swabs retais e realizado o isolamento seletivo, seguido da Reação de Polimerização em Cadeia (PCR) multiplex para identificar espécies e genes de resistência. Não foram isolados enterococos contendo os genes vanA ou vanB. A porcentagem de enterococos variou de 1,5 por cento para E. casseliflavus e E. gallinarum VanC até 12,3 por cento (n=8) para Enterococcus faecalis. A totalidade dos isolados da espécie E. faecalis demonstrou sensibilidade aos antimicrobianos vancomicina, teicoplanina, ampicilina, gentamicina e estreptomicina. Os genes de virulência ace e esp foram prevalentes (100 por cento, 87.5 por cento). A análise em multilocus de repetições em tandem de número variável (MLVA) revelou diversidade no número de repetições entre os isolados de E. faecalis, que foi mais alta para espC, efa5 e efa6. Foram identificados seis diferentes genotipos de MVLA, divergindo daqueles já descritos em humanos. Os genotipos foram ainda agrupados em dois genogrupos, demonstrando especificidade de hospedeiro para as espécies Cebus apella ou Callithrix penicillata. Concluindo, não foram isoladas linhagens de enterococos contendo os genes vanA ou vanB colonizando as espécies de primatas analisadas. O presente estudo demonstrou que os isolados de E. faecalis obtidos...