Sepsis neonatal temprana, incidencia y factores de riesgo asociados en un hospital público del occidente de México / Early neonatal sepsis, incidence and associated risk factors in a public hospital in western Mexico

Background: Early neonatal sepsis (EOS) is the second leading cause of death in the first week of life. Epidemiology differs in developed and developing countries. Aim: To describe the epidemiology of EOS among newborn patients in a public hospital in western Mexico. Methods: A prospective cohort study was performed in newborns of Nuevo Hospital Civil de Guadalajara "Dr. Juan I Menchaca". EOS was diagnosed with blood cultures or cultures of cerebrospinal fluid within the first 72 h of life. We analyzed risk factors (RF) by multivariate analysis with logistic regression. Results: We identified an EOS incidence of 4.7 events per 1,000 live births. Seventy two percent of the isolated bacteria were gram negative bacilli. Factors associated with EOS were maternal age ≤ 15 years (OR 3.50; 95% CI 1.56-7.85), rupture of membranes > 18 h (OR 2.65; 95% CI 1.18-5.92), maternal fever (OR 6.04; 95% CI 1.54-23.6), birth weight ≤ 2,500 g (OR 4.82; 95% CI 2.38-9.75) and gestational age < 37 weeks (OR 3.14; 95% CI 1.58-6.22). Conclusions: In addition to the RF known for EOS an independent association was observed with maternal age ≤ 15 years.

Introducción: La sepsis neonatal temprana (SNT) es la segunda causa de muerte en la primer semana de vida; la epidemiología difiere en países desarrollados y en vías de desarrollo. Objetivo: Describir la epidemiología de SNT en recién nacidos (RN) de un hospital público del occidente de México. Material y Métodos: Estudio de cohorte prospectivo en RN del Nuevo Hospital Civil de Guadalajara "Dr. Juan I Menchaca". Se diagnosticó SNT con cultivos de sangre o líquido cefalorraquídeo en las primeras 72 h de vida. Se indagaron factores de riesgo (FR) mediante análisis multivariado con regresión logística. Resultados: La incidencia de SNT fue de 4,7 eventos por 1.000 RN vivos. El 72% de las bacterias aisladas correspondió a bacilos gramnegativos. Los factores asociados a SNT fueron la edad materna ≤ 15 años (OR 3,50; IC 95% 1,56-7,85), ruptura de membranas > 18 h (OR 2,65; IC 95% 1,18-5,92), fiebre materna (OR 6,04; IC 95%1,54-23,6), peso al nacimiento ≤ 2.500 g (OR 4,82; IC 95% 2,38-9,75) y edad gestacional < 37 semanas (OR 3,14; IC 95% 1,58-6,22). Conclusiones: Además de los FR ya conocidos para SNT se observó asociación independiente con edad materna ≤ 15 años.

COMPARISON BETWEEN AUTOMATED SYSTEM AND PCR-BASED METHOD FOR IDENTIFICATION AND ANTIMICROBIAL SUSCEPTIBILITY PROFILE OF CLINICAL Enterococcus spp / Comparação entre o sistema automatizado e PCR na identificação e susceptibilidade de isolados clínicos de Enterococcus spp

Enterococci are increasingly responsible for nosocomial infections worldwide. This study was undertaken to compare the identification and susceptibility profile using an automated MicrosScan system, PCR-based assay and disk diffusion assay of Enterococcus spp. We evaluated 30 clinical isolates of Enterococcus spp. Isolates were identified by MicrosScan system and PCR-based assay. The detection of antibiotic resistance genes (vancomycin, gentamicin, tetracycline and erythromycin) was also determined by PCR. Antimicrobial susceptibilities to vancomycin (30 µg), gentamicin (120 µg), tetracycline (30 µg) and erythromycin (15 µg) were tested by the automated system and disk diffusion method, and were interpreted according to the criteria recommended in CLSI guidelines. Concerning Enterococcus identification the general agreement between data obtained by the PCR method and by the automatic system was 90.0% (27/30). For all isolates of E. faecium and E. faecalis we observed 100% agreement. Resistance frequencies were higher in E. faecium than E. faecalis. The resistance rates obtained were higher for erythromycin (86.7%), vancomycin (80.0%), tetracycline (43.35) and gentamicin (33.3%). The correlation between disk diffusion and automation revealed an agreement for the majority of the antibiotics with category agreement rates of > 80%. The PCR-based assay, the van(A) gene was detected in 100% of vancomycin resistant enterococci. This assay is simple to conduct and reliable in the identification of clinically relevant enterococci. The data obtained reinforced the need for an improvement of the automated system to identify some enterococci.

Os enterococos são cada vez mais responsáveis por infecções hospitalares em todo o mundo. Este estudo foi realizado para comparar a identificação e perfil de suscetibilidade entre o sistema automatizado MicrosScan e a técnica molecular de PCR em espécies de Enterococcus spp. Foram avaliados 30 isolados clínicos de Enterococcus spp. Os isolados foram identificados pelo sistema MicrosScan® e pela técnica de PCR. A detecção de genes de resistência a antibióticos (vancomicina, gentamicina, tetraciclina e eritromicina) foi determinada por PCR. Suscetibilidades antimicrobianas à vancomicina (30 µg), gentamicina (120 µg), tetraciclina (30 µg) e eritromicina (15 µg), foram testados pelos métodos automatizados e pelo disco difusão, de acordo com as orientações do CLSI. No que diz respeito à identificação de Enterococcus em geral entre os dados obtidos pelo método de PCR e pelo sistema automático foi de 90,0% (27/30). Para todos os isolados de E. faecium e E. faecalis observamos concordância de 100%. Freqüências de resistência foi maior em E. faecium do que em E. faecalis. As taxas de resistência obtidas foi maior para eritromicina (86,7%), vancomicina (80,0%), tetraciclina (43,35%) e gentamicina (33,3%). A correlação entre a técnica de disco difusão e automação revelou-se de acordo para maioria dos antibióticos com taxas > 80%. O gene van(A) foi detectado em 100% dos Enterococcus resistentes á vancomicina. O ensaio baseado em PCR é de simples realização e de confiança para identificação de enterococos clinicamente relevantes. Os dados obtidos reforçam a necessidade de melhoria no sistema automatizado para identificar alguns enterococos.

Prevalence and phenotypic characterization of Enterococcus spp. isolated from food in Brazil

We evaluated the frequency of enterococci from food and found 95.2% of positivity, being E. faecium and E. faecalis the most frequent species. High-level streptomycin resistance was observed, as well as gelatinase and hemolysis activity, showing the potential role of environmental strains as reservoir of virulence and resistance traits.

Fatal meningitis caused by vancomycin-resistant enterococci: Report of two cases from south India.

Vancomycin-resistant enterococci rarely cause meningitis and present a therapeutic challenge. Antimicrobial susceptibility testing was done for strains of Enterococcus species isolated from CSF samples of patients with meningitis by phenotypic methods. Multiplex polymerase chain reaction was performed to determine the genetic basis of vancomycin resistance of such isolates. We report here two cases of enterococcal meningitis caused by vancomycin-resistant Enterococcus species. One of the isolates was identified as Enterococcus faecalis and the other as Enterococcus gallinarum. We also report the simultaneous presence of vanC1 and vanA resistance genes in the strain of E. gallinarum. To the best of our knowledge, this is the first report of vanA resistance gene in an isolate of E. gallinarum from the Indian subcontinent. This is also the first Indian report of vancomycin-resistant Enterococcus causing meningitis.

Risk factors for vancomycin-resistant enterococci colonisation in critically ill patients

Vancomycin-resistant enterococci (VRE) are important hospital pathogens and have become increasingly common in patients admitted to the intensive care unit (ICU). To determine the incidence and the risk factors associated with VRE colonisation among ICU patients, active surveillance cultures for VRE faecal carriages were carried out in patients admitted to the ICU of the University Hospital of Uberlândia, Minas Gerais, Brazil. Risk factors were assessed using a case-control study. Seventy-seven patients (23.1 percent) were found to be colonised with vanC VRE and only one patient (0.3 percent) was colonised with vanA VRE. Independent risk factors for VRE colonisation included nephropathy [odds ratio (OR) = 13.6, p < 0.001], prior antibiotic use (OR = 5.5, p < 0.03) and carbapenem use (OR = 17.3, p < 0.001). Our results showed a higher frequency (23.1 percent) of Enterococcus gallinarum and Enterococcus casseliflavus, species that are intrinsically resistant to low levels of vancomycin (vanC), without an associated infection, associated with prior antibiotic use, carbapenem use and nephropathy as comorbidity. This study is the first to demonstrate the risk factors associated with vanC VRE colonisation in ICU hospitalised patients. Although vanA and vanB enterococci are of great importance, the epidemiology of vanC VRE needs to be better understood. Even though the clinical relevance of vanC VRE is uncertain, these species are opportunistic pathogens and vanC VRE-colonised patients are a potential epidemiologic reservoir of resistance genes.

Prevalence and antimicrobial resistance pattern of multidrug-resistant enterococci isolated from clinical specimens.

Purpose: Vancomycin-resistant enterococci (VRE) pose an emerging problem in hospitals worldwide. The present study was undertaken to determine the occurrence, species prevalence, antibacterial resistance, and phenotypic and genetic characteristics of VRE isolated in Riyadh hospitals, KSA. Materials and Methods: Two hundred and six isolates of enterococcal species were obtained from clinical samples. The antibiotic susceptibility of isolates and minimum inhibitory concentration (MIC) tests for vancomycin and teicoplanin were determined. Molecular typing of VRE isolates was carried out by using pulsed field gel electrophoresis (PFGE) and the resistance genotype was determined by polymerase chain reaction (PCR). Results: VRE accounted for 3.9% of the isolates and were detected mostly in urine, wound and blood specimens isolated from ICU, internal medicine and surgical wards. All strains were identified to species level and were found to consist of E. faecalis (69.2%), E. faecium (11.3%), E. avium (2.1%), E. hirae (0.8%), E. casseliflavus (1.3%) and E. gallinarum (1.3%) species. According to the susceptibility data obtained, 8 (3.9%) out of 206 isolates were found to be VRE (MICs > 32 μg/ml). The vanA, vanB and vanC gene fragments of E. faecalis, E. faecium and E. gallinarum were amplified from isolates and were detected. PFGE patterns of the VRE isolates revealed homogenous patterns with dominant clone suggesting that the strains intrinsic resistance is independent. Conclusions: This study shows an emergence of VRE along with increased rate of multidrug-resistant enterococci in the area of the study. Regular surveillance of antimicrobial susceptibilities should be done regularly and the risk factors should be determined.

Trends in antimicrobial resistance among clinical isolates of enterococci in a Brazilian tertiary hospital: a 4-year study / Evolução da resistência aos antimicrobianos entre isolados clínicos de enterococos em um hospital terciário brasileiro: um estudo de 4 anos

INTRODUCTION: In the past two decades members of the genus Enterococcus have emerged as important nosocomial pathogens worldwide. This study prospectively analyzed the distribution of species and trends in antimicrobial resistance among clinical isolates of enterococci in a Brazilian tertiary hospital from 2006-2009. METHODS: Enterococcal species were identified by conventional biochemical tests. The antimicrobial susceptibility profile was performed by disk diffusion in accordance with the Clinical and Laboratory Standards Institute (CLSI). A screening test for vancomycin was also performed. Minimal inhibitory concentration (MIC) for vancomycin was determined using the broth dilution method. Molecular assays were used to confirm speciation and genotype of vancomycin-resistant enterococci (VRE). RESULTS: A total of 324 non-repetitive enterococcal isolates were recovered, of which 87 percent were E. faecalis and 10.8 percent E. faecium. The incidence of E. faecium per 1,000 admissions increased significantly (p < 0.001) from 0.3 in 2006 to 2.3 in 2009. The VRE rate also increased over time from 2.5 percent to 15.5 percent (p < 0.001). All VRE expressed high-level resistance to vancomycin (MIC >256µg/ mL) and harbored vanA genes. The majority (89.5 percent) of VRE belonged to E. faecium species, which were characteristically resistant to ampicillin and quinolones. Overall, ampicillin resistance rate increased significantly from 2.5 percent to 21.4 percent from 2006-2009. Resistance rates for gentamicin, chloramphenicol, tetracycline, and erythromycin significantly decreased over time, although they remained high. Quinolones resistance rates were high and did not change significantly over time. CONCLUSIONS: The data obtained show a significant increasing trend in the incidence of E. faecium resistant to ampicillin and vancomycin.

INTRODUÇÃO: Nas últimas duas décadas, os enterococos emergiram como importantes patógenos nosocomiais no mundo inteiro. Neste estudo, foi analisada a distribuição das espécies e a evolução da resistência aos antimicrobianos entre isolados clínicos de enterococos obtidos em um hospital terciário, no período de 2006 a 2009. MÉTODOS: As espécies foram identificadas por testes bioquímicos convencionais e o perfil de sensibilidade foi determinado pelo método de disco difusão. A sensibilidade à vancomicina foi também determinada pela triagem em agar e pela concentração inibitória mínima (CIM). Testes moleculares foram utilizados para confirmar as espécies e determinar os genótipos dos enterococos resistentes à vancomicina (VRE). RESULTADOS: Foram analisadas 324 amostras de enterococos, sendo 87 por cento E. faecalis e 10,8 por cento E. faecium. A incidência de E. faecium por 1.000 pacientes internados aumentou significativamente (p < 0,001) de 0,3 em 2006 para 2,3 em 2009. A taxa de VRE também aumentou significativamente de 2,5 por cento para 15,5 por cento (p < 0,001). Todos os VRE apresentaram genótipo VanA e CIM >256µg/mL para vancomicina. A maioria (89,5 por cento) dos VRE pertencia à espécie E. faecium e foram resistentes à ampicilina e quinolonas. Foi observado um aumento significativo na taxa de resistência à ampicilina, de 2,5 por cento (2006) para 21,4 por cento (2009). As taxas de resistência para gentamicina, cloranfenicol, tetraciclina e eritromicina diminuíram significativamente no período do estudo. Para as quinolonas, as taxas de resistência foram elevadas não alteraram significativamente, no período do estudo. CONCLUSÕES: Os resultados do presente estudo mostram um aumento significativo na incidência de E. faecium resistentes à ampicilina e vancomicina.

Enterococcus gallinarum meningitis in an immunocompetent host: a case report / Meningite por Enterococcus gallinarum em paciente sem imunodepressão: relato de caso

We describe a rare case of a 53-year-old man with a long history of alcohol abuse, with Enterococcus gallinarum meningitis, an organism that rarely causes human infection and is primarily found in the gastrointestinal tract of poultry. The patient improved with high-dose ampicillin and gentamicin therapy. To our knowledge, this is the first Brazilian reported case of E. gallinarum meningitis and probably the first case described in an immunocompetent host.

Descrevemos caso raro de paciente de 53 anos com história de alcoolismo prévio, com meningite por Enterococcus gallinarum, um organismo que raramente causa infecções em humanos e é encontrado principalmente no trato gastrointestinal de aves. O paciente teve melhora importante após início de tratamento intravenoso com ampicilina e gentamicina combinados. Para o nosso conhecimento, este é o primeiro caso relatado de meningite por E. gallinarum no Brasil e possivelmente o primeiro caso descrito em paciente sem imunodepressão.

PCR-RFLP of 16S ribosomal DNA to confirm the identification of Enterococcus gallinarum and Enterococcus casseliflavus isolated from clinical and food samples / PCR-RFLP do 16S DNA ribossomal para confirmar a identificação de Enterococcus gallinarum e Enterococcus casseliflavus isolados de amostras clínicas e alimentares

INTRODUCTION: This study aimed to confirm the identification of Enterococcus gallinarum and Enterococcus casseliflavus isolated from clinical and food samples by PCR-RFLP. METHODS: Fifty-two strains identified by conventional biochemical exams were submitted to PCR amplification and digested with HinfI. Only 20 (38.5 percent) of the 52 strains showed a DNA pattern expected for E. gallinarum and E. casseliflavus. RESULTS: Analysis of the results of this study showed that E. gallinarum and E. casseliflavus are occasionally erroneously identified and confirmed the potential application of 16S rDNA analysis for accurate identification of these species. CONCLUSIONS: A correct identification is important to distinguish between intrinsic and acquired vancomycin resistance.

INTRODUÇÃO: O objetivo deste estudo foi confirmar a identificação de amostras clínicas e alimentos de Enterococcus gallinarum e Enterococcus casseliflavus por PCR-RFLP. MÉTODOS: Cinquenta e duas cepas identificadas por exames bioquímicos convencionais foram submetidos a amplificação por PCR e digestão com HinfI. Apenas 20 (38,5 por cento) das 52 amostras apresentaram um padrão de DNA esperado E. gallinarum e E. casseliflavus. RESULTADOS: Analise dos resultados deste estudo demonstraram que, algumas vezes E. gallinarum e E. casseliflavus são erroneamente identificados e confirmaram a potencial aplicação da análise do 16S rDNA para identificação exata destas espécies. CONCLUSÕES: A correta identificação é importante a fim de distinguir entre resistência intrínseca e adquirida à vancomicina.

Cross-transmission of vancomycin-resistant Enterococcus in patients undergoing dialysis and kidney transplant

The objective of this study was to investigate the occurrence of vancomycin-resistant Enterococcus (VRE) cross-transmission between two patient groups (long-term dialysis and kidney transplant patients). Molecular typing, by automated ribotyping with the RiboPrinter Microbial Characterization System (Qualicon, USA), was used to analyze VRE isolates from 31 fecal samples of 320 dialysis patients and 38 fecal samples of 280 kidney transplant patients. Clonal spread of E. faecalis and E. casseliflavus was observed intragroup, but not between the two groups of patients. In turn, transmission of E. gallinarum and E. faecium between the groups was suggested by the finding of vancomycin-resistant isolates belonging to the same ribogroup in both dialysis and transplant patients. The fact that these patients were colonized by VRE from the same ribogroup in the same health care facility provides evidence for cross-transmission and supports the adoption of stringent infection control measures to prevent dissemination of these bacteria.

RFLP analysis of a PCR-amplified fragment of the 16S rRNA gene as a tool to identify Enterococcus strains

Restriction fragment length polymorphism (RFLP) analysis of a PCR-amplified fragment of the 16S rRNA gene was performed on reference strains belonging to 21 different enterococcal species and on 75 Enterococcus isolates recovered from poultry meat, pasteurised milk and fresh cheese. PCR amplification generated a 275 bp fragment, which was digested with three restriction endonucleases (DdeI, HaeIII, HinfI). The strains were divided into five groups (groups A-E) on the basis of their restriction patterns. Five biochemical tests (arabinose, arginine, manitol, methyl-â-D-glucopyranoside and raffinose) were then performed in addition to RFLP analysis to narrow the identification of enterococcal strains to the species level. PCR-RFLP, in conjunction with the selected biochemical tests, allowed the precise identification of the 21 species of Enterococcus included in the present study. This proposed method is relatively simple and rapid and can be useful as an adjunct tool for accurate identification of Enterococcus.

Evaluation of the automated system Vitek2 for identification and antimicrobial susceptibility testing of Brazilian Gram-positive cocci strains

Automated instruments offer many advantages for clinical laboratories. Nevertheless, they can have problems identifying and determining susceptibilities of some pathogens. Vitek® 2 (bioMérieux) is an automated system that was recently introduced to Brazil. We evaluated the performance of this equipment for Brazilian isolates that had been characterized using reference identification and antimicrobial susceptibility testing methods. Ninety-nine strains of Gram-positive cocci from a local reference center collection were analyzed, consisting of 50 coagulasenegative Staphylococcus (CoNS) and 49 Enterococcus and related species. Vitek® 2 correctly identified 79.8 percent (79/99) of the isolates. Oxacillin resistance was detected in 76 percent (19/25) of resistant S. epidermidis strains and in 88 percent (22/25) of other resistant CoNS species strains. Vancomycin resistance was detected in 100 percent (20/20) of resistant Enterococcus and related species strains. Vitek® 2 performed very well for the identification of S. epidermidis and non-epidermidis staphylococci, and for the detection of vancomycin resistance in Enterococcus and related species. However, the system needs improvement in order to provide reliable results for the characterization of some CoNS species, identification of Enterococcus and related species and for detecting oxacillin resistance in CoNS.

Microbiological and Epidemiological Characteristics of Vancomycin-dependent Enterococci / 대한진단검사의학회지

BACKGROUND: Vancomycin-dependent enterococci (VDE) are clinically equivalent to vancomycin-resistant enterococci (VRE), but more difficult to detect. This study was purposed to characterize VDE microbiologically and epidemiologically. METHODS: The patients from whom VDE were detected from April 2007 to March 2008 were investigated. For available isolates, minimal inhibitory concentrations (MICs) of and the levels of dependence on vancomycin and teicoplanin were measured by E test (AB Biodisk, Sweden), and a test for reversion of VDE to non-dependent VRE (NDVRE) and pulsed field gel electrophoresis (PFGE) were performed. Patients' demographic and clinical findings were reviewed via electronic medical records. RESULTS: VDE were recovered from 6 (2.2%) of 272 patients carrying VRE during this study period. All patients were already colonized or infected by VRE and treated with vancomycin for 13 to 107 days. VDE were isolated from pleural fluid (one), urine (four), and stool (one). All isolates carried vanA with vancomycin MICs of >256 microgram/mL, but two of them had intermediate susceptibilities to teicoplanin. Because 4 VDE isolates were reverted to NDVRE with single passage, vancomycin dependence was measurable for only two isolates as equal and above 0.064 and 0.5 microgram/mL respectively, and was reverted after 5 and 7 passages, respectively. Six VDE isolates showed no related clones in PFGE analysis, and 3 of 4 available pairs of initial VRE isolates and subsequent VDE isolates were identical clones. CONCLUSIONS: VDE were not rare and seemed to emerge independently from VRE with a prolonged use of vancomycin. Vancomycin-dependence was reverted within several passages.

Antimicrobial resistance profiles of enterococci isolated from poultry meat and pasteurized milk in Rio de Janeiro, Brazil

The enterococci are important nosocomial pathogens with a remarkable capacity of expressing resistance to several antimicrobial agents. Their ubiquitous nature and resistance to adverse environmental conditions take account for their ability to colonize different habitats and for their potential for easy spreading through the food chain. In the present study we evaluated the distribution of species and antimicrobial susceptibility among enterococcal isolates recovered from food obtained in retail stores in Rio de Janeiro, Brazil. The following species were identified among 167 isolates obtained from poultry meat and 127 from pasteurized milk: Enterococcus faecalis (62.6 percent), E. casseliflavus (17.3 percent), E. durans (6.5 percent), E. gallinarum (3.0 percent), E. gilvus (2.4 percent), E. faecium (2.0 percent), E. hirae (1.4 percent), and E. sulfureus (1.0 percent). The overall percentages of antimicrobial resistant isolates were: 31.2 percent to tetracycline, 23.8 percent to erythromycin, 11.3 percent to streptomycin, 4.3 percent to chloramphenicol, 3.9 percent to gentamicin, 1.4 percent to norfloxacin, 1.1 percent to imipenem, 0.7 percent to ciprofloxacin, nitrofurantoin, and penicillin, and 0.4 percent to ampicillin. Intermediate resistance was detected in frequencies varying from 0.5 percent for linezolid to 58.2 percent for erythromycin. None of the isolates showed resistance to glycopeptides. High-level resistance to aminoglycosides was observed in 13.1 percent of the isolates. Multiresistance was observed in E. faecalis, E. casseliflavus, E. faecium, E. gallinarum, E. durans and E. gilvus.

Low prevalence of vancomycin resistant enterococci colonization in intensive care patients in a Brazilian teaching hospital

Vancomycin-resistant enterococci (VRE) are important pathogens involved in nosocomial infections. Colonization precedes infection and the number of colonized individuals is about 10 times higher than the number of infected patients. We examined VRE colonization in two intensive care units from October 2003 to June 2004. Perirectal swab specimens were obtained from all patients, starting on the 5th day after admission, and then weekly. A total of 249 swabs were obtained from 112 patients. Nine patients had VRE-positive swabs, giving a positive rate of 8.0 percent. The rate of patients colonized by V-R E. faecalis was 1.8 percent (n=2), 4.5 percent by V-R E. gallinarun (n=5) and 1.8 percent by V-R E. casseliflavus (n=2). No V-R E. faeciun was isolated. None of the patients that had been colonized by VRE were found to be infected by these pathogens. In summary, a low prevalence of colonization by VRE was found in our institution. Only a structured surveillance program, based on active searching, was able to detect this low number of cases.

In vitro resistance to human platelet microbicidal protein among urethral staphylococcal and enterococcal isolates with its correlation with prostatitis.

The study was carried out to test the in vitro activity of human platelet microbicidal protein (hPMP) on most commonly isolated urethral pathogens and compare the same with clinical isolates from cases of chronic prostatitis (CP). Urethral isolates of Staphylococcus aureus (n=19), coagulase negative staphylococci (n=40) and Enterococcus faecalis (n=16) from patients with or without CP were tested. The hPMP susceptibility of bacterial strains was determined by exposing bacterial cells to serial dilutions of hPMP. A significantly higher proportion of CP-strains of coagulase negative staphylococci (91.3% vs 5.88%) was resistant to hPMP than was that of non-CP strains (P S.aureus studied, 77.8% were considered resistant to the bactericidal action of hPMP. All nine CP-strains of E.faecalis were highly resistant to hPMP. Most non-CP urethral isolates of S.aureus, coagulase negative staphylococci and E.faecalis were susceptible to the bactericidal action of hPMP, while CP isolates of all species were significantly more resistant to hPMP. Data from the present study may have significant implications in understanding the pathogenesis of CP.

Species prevalence and antimicrobial susceptibility of enterococci isolated in a tertiary care hospital of North India.

The present prospective study was carried out to determine the species distribution and antimicrobial susceptibilities of enterococci isolated from clinical samples in a tertiary care hospital of North India. Enterococcus species isolated from blood, urine, pus, sterile fluids and the hospital environment from October 2003 to January 2004 were identified by standard biochemical tests. Antimicrobial susceptibility testing was performed by the disk diffusion method as per NCCLS guidelines. Out of a total of 105 Enterococcus species recovered during the study period, E. faecium (42.90%) and E. faecalis (40.00%) constituted the predominant isolates. Enterococcus faecium was the commonest blood culture isolate while E. faecalis predominated pus and urine samples. Other species isolated were E. mundtii, E dispar, E. durans, E. avium, E. raffinosus and E. gallinarum. High-level aminoglycoside resistance was detected in 73.3% of isolates. Resistance to vancomycin, teicoplanin and linezolid was not detected. Prevalence of a wide variety of Enterococcus species in clinical samples together with their variable antimicrobial susceptibility patterns emphasizes the need for routinely carrying out detailed speciation and in vitro susceptibility testing of enterococcal isolates in the clinical bacteriology laboratory.

Enterococcus gallinarum carrying the vanA gene cluster: first report in Brazil

In 2000, Enterococcus faecalis resistant to vancomycin was first reported at a tertiary hospital in Porto Alegre, southern Brazil. The resistance spread to other hospitals and surveillance programs were established by hospital infection committees to prevent the spread of vancomycin-resistant enterococci. In February 2002, an isolate initially identified at the genus level as Enterococcus was obtained by surveillance culture (rectal swab) from a patient admitted to a hospital for treatment of septic arthritis in the shoulder. The isolate proved to be resistant to vancomycin by the disc diffusion method and confirmed by an E-test resulting in a minimal inhibitory concentration of > ou = 256 µg/ml. This isolate was sent to a reference laboratory (Laboratório Especial de Bacteriologia e Epidemiologia Molecular, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, USP) for further study and proved to be an E. gallinarum by the polymerase chain reaction (PCR) using specific primers for the species. Due to the phenotype of unusually high vancomycin resistance, the isolate presumably had the resistance genes (vanA and vanB) and this was confirmed by PCR, which indicated the presence of the vanA gene. A 10.8-kb Tn1546-related transposon was also identified by long-PCR. Interspecies transfer of the vancomycin-resistance gene from the donor E. gallinarum was performed in a successful conjugation experiment in vitro, using E. faecium GE-1 and E. faecalis JH22 as receptors. This is the first report of the detection of a vanA determinant naturally acquired by E. gallinarum in Brazil, indicating the importance of characterizing VRE by both phenotype and genotype methods.