Bacteriemia por Enterococcus faeciumresistente a daptomicina / Daptomycin-resistant enterococcus faecium bacteremia

La aparición de cepas de enterococos resistentes a daptomicina es un tema de preocupación clínica y epidemiológica en años recientes. Se presenta el caso de un paciente de 50 años con antecedente de artritis reumatoide e inmunosupresión crónica hospitalizado en contexto de neumonía viral por COVID-19, con sobreinfección bacteriana y choque séptico, en quien se documentó en tres oportunidades diferentes aislamientos de Enterococcus faecium vancomicino-resistente VAN A y B con falla terapéutica a daptomicina, por deterioro clínico y persistencia de hemocultivos positivos. Se inició manejo con linezolid con control de la infección, negativización de hemocultivos y evolución clínica satisfactoria. Se realiza reporte del presente caso para dar a conocer la aparición de enterococos resistentes a daptomicina, la cual es una creciente preocupación epidemiológica, con el fin de realizar identificación temprana, prevenir falla terapéutica y poder conocer la epidemiología local

n recent years, the emergence of daptomycin-resistant enterococcus strains is a growing clinical and epidemio-logical topic of concern. We report the case of a 50 year old patient with an antecedent of rheumatoid arthritis and chronic immunosuppression hospitalized in the con-text of COVID-19 pneumonia with bacterial co-infection and septic shock in which a three different moments an isolate of a "vancomycin-resistant enterococcus faecium"(VRE) Van A and B that presented therapeutic failure with daptomycin was documented after clinical deterioration and persistence of positive blood cultures. Linezolid was initiated, with clinical recovery and negativization of blood cultures following the change in antibiotic treatment. This case is reported in order to expose the ever growing con-cern of daptomycin-resistant enterococcus strains in or-der to prevent therapeutic failure, make early identifica-tion and get to know the local epidemiological status.

E.faecium QH06 alleviates TNBS-induced colonic mucosal injury in rats / 南方医科大学学报

OBJECTIVE@#To investigate the effect of Enterococcus faecium QH06 on TNBS-induced ulcerative colitis (UC) in rats and explore the mechanisms in light of intestinal flora and intestinal immunity.@*METHODS@#Thirty-six male Wistar rats were randomized equally into control group, UC model group, and E.faecium QH06 intervention group. The rats in the latter two groups were subjected to colonic enema with 5% TNBS/ethanol to induce UC, followed by treatment with intragastric administration of distilled water or E.faecium QH06 at the dose of 0.21 g/kg. After 14 days of treatment, the rats were examined for colon pathologies with HE staining. The mRNA and protein expression levels of IL-4, IL-10, IL-12, and IFN-γ in the colon tissues were detected using RT-qPCR and ELISA, and the expression of TLR2 protein was detected with immunohistochemistry and ELISA. Illumina Miseq platform was used for sequencing analysis of the intestinal flora of the rats with bioinformatics analysis. The correlations of the parameters of the intestinal flora with the expression levels of TLR2 and cytokines were analyzed.@*RESULTS@#The rats with TNBS- induced UC showed obvious weight loss (P < 0.01) and severe colon tissue injury with high pathological scores (P < 0.01). The protein expression levels of IFN-γ, IL-12, and TLR2 (P < 0.01) and the mRNA expression levels of IFN-γ, IL-12 and IL-10 (P < 0.05) were significantly increased in the colon tissues of the rats with UC. Illumina Miseq sequence analysis showed that in UC rats, the Shannon index (P < 0.05) ACE (P < 0.01)and Chao (P < 0.05) index for the diversity of intestinal flora both decreased with a significantly increased abundance of Enterobacteriaceae (P < 0.01) and a lowered abundance of Burkholderiaceae (P < 0.05). Compared with the UC rats, the rats treated with E. faecium QH06 showed obvious body weight gain (P < 0.05), lessened colon injuries, lowered pathological score of the colon tissue (P < 0.05), decreased protein expressions of IFN- γ, IL- 12, and TLR2 and mRNA expressions of IFN- γ and IL-12 (P < 0.01 or 0.05), and increased protein expressions of IL- 4 (P < 0.05). The Shannon index ACE (P < 0.05) and Chao (P < 0.05) index of intestinal microflora were significantly increased, the abundance of Enterobacteriaceae was lowered and that of Burkholderiaceae and Rikenellaceae was increased in E.faecium QH06- treated rats (P < 0.01 or 0.05). Correlation analysis showed that IFN-γ was positively correlated with the abundance of Enterobacteriaceae, and IFN-γ was negatively correlated with the abundance of Prevotellaceae, Desulfovibrionaceae, norank_o_Mollicutes_RF39 and Clostridiales_vadinBB60_group; TLR2 was negatively correlated with Clostridiales_vadinBB60_group, norank_o_Mollicutes_RF39 and Prevotellaceae.@*CONCLUSION@#E.faecium QH06 can alleviate TNBS-induced colonic mucosal injury in rats, and its effect is mediated possibly by increasing the abundance of SCFA-producing bacteria such as Prevotellaceae and inhibiting abnormal immune responses mediated by TLR2.

Staphylococcus aureus and Enterococcus faecium isolated from pigeon droppings (Columba livia) in the external environment close to hospitals

ABSTRACT Background: Domestic pigeons carry pathogens in their droppings, posing a potential public health problem. Methods: The phenotypic and genotypic antimicrobial resistances of Staphylococcus aureus and Enterococcus faecium in the feces of urban pigeons near hospitals with intensive care units were measured. Results: Twenty-nine samples showed Enterococcus growth, whereas one was positive for S. aureus. The S. aureus isolate was sensitive to the antibiotics tested via antibiogram, however resistance genes were identified. E. faecium isolates showed phenotypic resistance to gentamicin, erythromycin, and ciprofloxacin. Conclusions: Antimicrobial profiles harmful to health were demonstrated in bacterial pathogens isolated from the external environment of hospitals.

Analysis of the clinical characteristics and susceptibility factors for bloodstream infections of Enterococcus faecium in children / 中华实用儿科临床杂志

Objective:To explore the clinical characteristics and susceptibility factors for bloodstream infections (BSI) of Enterococcus faecium in children, thus providing guidance for clinical prevention and treatment. Methods:Clinical data of 18 children with BSI of Enterococcus faecium treated in the West China Second University Hospital, Sichuan University from January 2011 to October 2019 were collected.Their underlying diseases, predisposing factors, clinical information, etiological data, treatment and prognosis were retrospectively analyzed. Results:Sixteen of 18 children (88.9%) with BSI of Enterococcus faecium had underlying diseases, including intestinal diseases or intestinal surgery, preterm infants, hematological diseases, congenital heart disease, biliary atresia and urinary system malformations.Seventeen of 18 children (94.4%) had predisposing factors, including broad-spectrum antibiotics exposure, Intensive Care Unit (ICU) admission, central venous catheterization, malnutrition, abdominal distension or intestinal infection, granulopenia or agranulocyte, mechanical ventilation.The underlying diseases and susceptible factors varied with the age of patients.The most common predisposing factors of all-aged patients with BSI of Enterococcus faecium were broad-spectrum antibiotics exposure (76.5%) and central venous catheterization (52.9%). In patients with single infection of Enterococcus faecium, their C-reactive protein (CRP) was normal or slightly elevated and no children died.The incidences of respiratory failure, shock and severe sepsis in patients with single infection of Enterococcus faecium were 16.6%, 0% and 8.3% respectively, which were 33.3%, 33.3% and 66.7% respectively in 6 children (33.3%) with mixed infections, and one died.Vancomycin-resistant strains were not found, and Vancomycin treatment was effective that quickly relieved clinical symptoms. Conclusions:(1)Most of children with BSI of Enterococcus faecium have underlying diseases and susceptible factors.The most common predisposing factors of all-aged patients with BSI of Enterococcus faecium are broad-spectrum antibiotics exposure and central venous catheterization.(2)The pathogenicity of Enterococcus faecium is low, and the clinical manifestations of BSI are atypical.(3)There is a high rate of mixed infections of Enterococcus faecium, and the severity and mortality of the mixed infected children are much higher than those with single infection.(4)No Vancomycin-resistant strains are found, and vancomycin treatment is effective that quickly relieves clinical symptoms.(5)Rational use of antibiotics and strict management of central venous cathete-rization are the most important preventive measures.

Determinacion de los factores de riesgo en sepsis por Enterococcus vancomicino resistente. Estudio de casos y controles en pacientes criticamente enfermos / Determination of risk factors in sepsis by Enterococcus vancomicyn resistant. Study of cases and control in criticall ill patients

Resumen Introducción: las infecciones causadas por Enterococcus resistente a Vancomicina (EVR) presentan mayor mortalidad en pacientes críticos, asociado a un aumento gradual en este patrón de resistencia, especialmente en el continente americano, por lo cual la adecuada terapia antimicrobiana empírica es fundamental para mejorar los desenlaces. Objetivo: determinar los factores de riesgo asociados al desarrollo de infección por EVR en pacientes sépticos en la Unidad de Cuidados Intensivos (UCI) del Hospital San José en Bogotá, Colombia. Métodos: Estudio descriptivo de casos y controles en pacientes sépticos ingresados a la UCI durante 2016 y 2017. Los casos se definieron como pacientes con infección por EVR y los controles los pacientes con infección por otro germen. Resultados: se incluyeron 32 pacientes con aislamiento de EVR y 96 controles. Los factores de riesgo asociados a infección por EVR fueron: nutrición parenteral (OR 15,7 IC 4,2-71,4), lavado peritoneal (OR 8,9 IC 3,2-24,8), cultivo polimicrobiano (OR 19,9 IC 6,0-83,4). La mortalidad fue 56,2% en casos y 33,3% en controles. Conclusiones: Los factores de riesgo hallados con mayor frecuencia fueron: múltiples lavados peritoneales, nutrición parenteral y cultivos polimicrobianos. Encontramos una correlación significativa en el uso de antibiótico empírico adecuado y la reducción en la mortalidad.

Summary Introduction: infections caused by Vancomycin-resistant Enterococcus (VRE) have higher mortality in critically ill patients, associated with increase in this pattern of resistance, especially in the Americas, which is why adequate empirical antimicrobial therapy is essential to improve outcomes Objective: to determine the risk factors associated with the development of infection by VRE in septic patients in the Intensive Care Unit (ICU) of San José Hospital in Bogotá, Colombia. Methods: Case-control study in septic patients admitted to the ICU during 2016 and 2017. The cases were defined as patients with VRE infection and the controls were patients with infection by another germ. Results: 32 patients with EVR isolation and 96 controls were included. The risk factors associated with infection by EVR were: parenteral nutrition (OR 15.7 IC 4.2-71.4), peritoneal lavage (OR 8.9 IC 3.2-24.8), polymicrobial culture (OR 19,9 IC 6.0-83.4). Mortality was 56.2% in cases and 33.3% in controls. Conclusions: The risk factors found most frequently were: multiple peritoneal lavage, parenteral nutrition and polymicrobial cultures. We found a significant correlation in the use of adequate empirical antibiotic and the reduction in mortality

Uso compasional de tigeciclina intraventricular en pacientes con ventriculitis causada por Enterococcus faecium. Reporte de seis casos pediátricos / Compassional use of Intraventricular Tigecycline in ventriculitis caused by Enterococcus faecium. Six pediatric cases

Introducción: La Tigeciclina es un fármaco de uso restringido en pediatría. El uso de este antibiótico por vía intraventricular es una decisión de uso compasivo en casos de bacterias altamente resistentes, y para los casos en que no exista otra alternativa. Caso clínico: Se presenta seis casos de pacientes con diagnóstico de ventriculitis a Enterococcus faecium que recibieron tigeciclina intraventricular con evolución bacteriológica exitosa. Se discute las dosis utilizadas por vía endovenosa e intraventricular. Conclusiones: el uso de este antibiótico por vía intraventricular puede ser una alternativa exitosa en casos de gérmenes altamente resistentes y cuando no exista otra alternativa terapéutica.

Introduction: Tigecycline is a drug of restricted use in pediatrics. The use of this antibiotic intraventricularly is a decision of compassive use in cases of highly resistant bacteria, and in cases where there is no other alternative. Clinical case: We present six cases of patients with a diagnosis of ventriculitis caused by Enterococcus faecium, who received intraventricular tigecycline with successful microbiological evolution. The doses used intravenously and intraventricularly are discussed. Conclusions: the use of this antibiotic intraventricularly can be a successful alternative in cases of highly resistant germs, when there are not alternative options. Key words: ventriculitis, Enterococcus faecium, tigecycline, intraventricula

Effect of Functional Feed Catfish (Clarias gariepinus) Oil and Probiotic Enterococcus faecium IS-27526 on Lipid Profile of Aged Atherogenic Female Cynomolgus Monkey (Macaca fascicularis)

@#Introduction: Catfish oil contains saturated and unsaturated fatty acids. Polyunsaturated fatty acid contributes to maintain the lipid profile. Enterococcus faecium IS-27526 is a novel indigenous probiotic of dadih origin. The aim of this study was to find out the effect of probiotic and catfish (Clarias gariepinus) based functional feed on lipid profile of aged female Cynomolgus monkey/CM (Macaca fascicularis) fed with atherogenic diet. Methods: Nine CMs were randomly devided into 3 groups, in single blind intervention study for 90 days , namely: A1= atherogenic diet as control positive, A2= probiotic, A3= probiotic + catfish oil after 45 days adaptation period. The average of CM’s bodyweight was in a range of 3 to 4 kg. The main ingredients of atherogenic diet were catfish flour, egg, sugar, egg yolk flour, and condiments. Serum lipid profile was assessed monthly by Cholesterol Oxidase-Peroxidase Aminoantipyrine (CHOP-POD) methods for cholesterol, LDL and HDL, while Glycerol-3-Phosphate Oxidase Peroxidase-Antiperoxidase (GPO-PAP) method for Triglyceride. Results: Atherogenic diet did not significantly affect appetite and bodyweight of the animals (p=0,12). The atherogenic diet significantly increased cholesterol and LDL level of animals (p=0.03 and p = 0.04) after 90 days administration . The CMs fed with probiotic E. faecium IS-27526 and catfish oil showed significant higher serum cholesterol (p=0,03) and LDL levels (p=0,04) compared to atherogenic diet alone and atherogic diet with probiotic E. faecium IS-27526. Probiotic E. faecium IS-27526 at 108 cfu/day stabilized the cholesterol and LDL level after 90 days administration compared with other groups of intervention. The TG and HDL of each group showed within normal range during the whole intervention study. Conclusion: Atherogenic feed significantly increased cholesterol and LDL level of CMs. Probiotic E. faecium IS-27526 showed a potential beneficial effect in maintaining lipid profile and could prevent dyslipidemia.

Frequency of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPRs) in non-clinical Enterococcus faecalis and Enterococcus faecium strains / Frequência das repetições palindrômicas curtas agrupadas e regularmente interespaçadas (CRISPRs) em cepas não-clínicas de Enterococcus faecalis e Enterococcus faecium

Abstract The fidelity of the genomes is defended by mechanism known as Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) systems. Three Type II CRISPR systems (CRISPR1- cas, CRISPR2 and CRISPR3-cas) have been identified in enterococci isolates from clinical and environmental samples. The aim of this study was to observe the distribution of CRISPR1-cas, CRISPR2 and CRISPR3-cas in non-clinical strains of Enterococcus faecalis and Enterococcus faecium isolates from food and fecal samples, including wild marine animals. The presence of CRISPRs was evaluated by PCR in 120 enterococci strains, 67 E. faecalis and 53 E. faecium. It is the first report of the presence of the CRISPRs system in E. faecalis and E. faecium strains isolated from wild marine animal fecal samples. The results showed that in non-clinical strains, the CRISPRs were more frequently detected in E. faecalis than in E. faecium. And the frequencies of CRISPR1-cas and CRISPR2 were higher (60%) in E. faecalis strains isolated from animal feces, compared to food samples. Both strains showed low frequencies of CRISPR3-cas (8.95% and 1.88%). In conclusion, the differences in the habitats of enterococcal species may be related with the results observe in distribution of CRISPRs systems.

Resumo A fidelidade dos genomas ​​é defendida por mecanismos conhecidos como sistemas de repetições palindrômicas curtas agrupadas e regularmente interespaçadas (CRISPRs). Três tipos de sistemas CRISPR II (CRISPR1-cas, CRISPR2 e CRISPR3-cas) têm sido identificados em cepas de enterococos isolados de amostras clínicas e ambientais. O objetivo deste estudo foi observar a distribuição dos CRISPR1-cas, CRISPR2 e CRISPR3-cas em cepas não-clínicas de Enterococcus faecalis e Enterococcus faecium isoladas de amostras alimentícias e fecais, incluindo animais marinhos selvagens. A presenca dos CRISPRs foi determinada por PCR em 120 cepas de enterococos, sendo 67 E. faecalis e 53 E. faecium. É o primeiro relato da presença do sistema CRISPRs nas estirpes E. faecalis e E. faecium isoladas de amostras fecais de animais marinhos selvagens. Os resultados mostraram que em cepas não-clínicas, os CRISPRs foram mais frequentemente detectados em E. faecalis do que em E. faecium. E as frequências de CRISPR1-cas e CRISPR2 foram maiores (60%) em cepas de E. faecalis isoladas de fezes de animais, quando comparadas à amostras de alimentos. Ambas as cepas apresentaram baixas freqüências de CRISPR3-cas (8,95% e 1,88%). Em conclusão, as diferenças nos habitats das espécies de enterococos podem estar relacionadas com os resultados observados na distribuição dos sistemas CRISPRs.

Evaluación cuantitativa de riesgos microbiológicos asociado con el consumo de agua del río Chicamocha en Boavita- Boyacá / Quantitative microbiological risks assessment associated with the consumption of water of Chicamocha river in Boavita-Boyacá

RESUMEN Los habitantes del sector El Terrenal, del municipio de Boavita-Boyacá, al no contar con el servicio de acueducto, utilizan las aguas del río Chicamocha, para la preparación de sus alimentos, limpieza y aseo personal. El objetivo de este estudio fue evaluar bacterias y protozoos presentes en el cuerpo de agua del río Chicamocha, a través de un análisis cuantitativo de riesgo microbiológico (ACRM). Para el desarrollo del trabajo, se realizaron cuatro muestreos, de abril a junio de 2016, en los márgenes y centro del río y un quinto muestreo de agua tratada con la planta Opuntia ficus-indica (Tuno). Se realizó el método de recuento en placa, usándose medios selectivos y diferenciales para el aislamiento de Escherichia coli, Salmonella, Enterococcus y la técnica de MacMaster, para la determinación de parásitos gastrointestinales (PGI). Para el análisis de la información, se emplearon los estadísticos no paramétricos de Kruskall-Wallis y U-Mann Whitney. Se determinó que, al menos, un lugar de muestreo es estadísticamente diferente a otro para Enterococcus faecium (P= 0,000) y los ooquistes de Entamoeba histolytica (P= 0,000). Por el contrario, para Escherichia coli no se observaron diferencias significativas entre las zonas, pero sí al menos una fecha de muestreo fue estadísticamente diferente (P= 0,026) a otra. En el agua tratada, se observó una disminución de los ooquistes de E. histolytica y un aumento en el porcentaje de E. faecium (P= 0,035).

ABSTRACT The habitants of the El Terrenal sector of the municipality of Boavita-Boyacá, have no access to the aqueduct service, and use water of the Chicamocha River for preparation of food, housework and personal hygiene. The aim of this study was to evaluate the presence of pathogenic bacteria and protozoa, present in the water body of the Chicamocha River, through a quantitative analysis of microbiological risk (ACRM). For the development of the work, four samples were conducted from April to June 2016, in the margins and center of the river, and a fifth sample of water treated with the plant Opuntia ficus-indica (Tuno). The method of plate count was performed, using selective and differential media for the isolation of Escherichia coli, Salmonella, Enterococcus and the MacMaster technique for the determination of gastrointestinal parasites (PGI). For the analysis of the information the nonparametric statistics of Kruskall Wallis and U-Mann Whitney were used. It was determined that at least one sampling site is statistically different from another for Enterococcus faecium (P = 0.000) and the oocysts of Entamoeba histolytica (P = 0.000). In contrast, for Escherichia coli no significant differences were observed between the zones, but if at least one sampling date was statistically different (P =0.026) to another. In the treated water there was a decrease in the oocysts of E. histolytica and an increase in the percentage of E. faecium (P= 0,035).

Resistencia a los β -lactámicos en enterococos / Resistance to β-lactams in enterococci

Enterococci are intrinsically resistant to several antimicrobial classes and show a great ability to acquire new mechanisms of resistance. Resistance to β-lactam antibiotics is a major concern because these drugs either alone or in combination are commonly used for the treatment of enterococcal infections. Ampicillin resistance, which is rare in Enterococcus faecium occurs in most of the hospital-associated Enterococcus faecium isolates. High-level resistance to ampicillin in E. faecium is mainly due to the enhanced production of PBP5 and/or by polymorphisms in the beta subunit of this protein. The dissemination of high-level ampicillin resistance can be the result of both clonal spread of strains with mutated pbp5 genes and resistance horizontal gene transfer.

Los enterococos son intrínsecamente resistentes a varias clases de antimicrobianos y presentan una gran capacidad para adquirir mecanismos de resistencia. La resistencia a los antibióticos p-lactámicos es preocupante porque estos fármacos solos o combinados se usan comúnmente para el tratamiento de las infecciones enterocócicas. La mayoría de los aislamientos hospitalarios de Enterococcus faecium presentan resistencia a la ampicilina, la cual es rara en Enterococcus faecalis. El alto nivel de resistencia a la ampicilina en E. faecium se debe principalmente a la hiperproducción de PBP5 y/o a polimorfismos en la subunidad beta de esta proteína. La propagación de esta resistencia puede deberse tanto a la diseminación clonal de cepas con genes pbp5 mutados como a la transferencia horizontal de genes.

Mass spectrometry of serum polypeptides of Enterococcus faecium bloodstream infection in mice / 解放军医学杂志

Objective To analyze the serum polypeptide fingerprints of Enterococcus faecium (E. faecium) bloodstream infection in mice at different time points using matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-FOF MS), screen differential polypeptide peaks, and create a corresponding diagnostic model and look for new markers. Methods Ninety ICR mice were used to establish E. faecium bloodstream infection model (including 10 mice as normal control group with PBS injection, and 80 mice as the infection group with 1/2 LD50 E. faecium injection in a volume of 0.1 ml/10 g by the intravenous tail). Blood samples were collected at 8 time points (1, 3, 6, 12, 24, 48, 96 and 128 h), and tested for IL-1α, IL-1β and IL-6 detection. Serum samples were then purified with weak cation magnetic beads, the serum polypeptide profiling was recognized by MALDI-TOF MS and BioExplorer software between infection group and normal control group. Amino acid sequences of the candidate polypeptides were identified by nano-liquid chromatography electrospray ionization-tandem mass spectrometry (nano-LC/ESI–MS/MS). Results After infected with E. faecium, the mice showed abnormal behaviors compared with the normal control group, including listless and reduced activity; IL-1α was down-regulated, while IL-1β and IL-6 were up-regulated. A total of 102 polypeptide peaks were detected by fingerprints analysis. Screening on the basis of 5 times difference in content in both infection group and control group, 8 polypeptides were higher in infection group than those in control group (P<0.01), and 9 polypeptides were lower in infection group than those in control group (P<0.01). Combining five polypeptide peaks of m/z 1227.4, 1512.9, 4509.7, 5007.3 and 7816.7, the corresponding diagnostic model was established with accuracy rate of 80%, specificity of 76.6% and sensitivity of 83.3%. Identified by secondary mass spectrometry, polypeptide peaks m/z 1227.4 and 5007.3 were the fragment of β2microglobulin and complement C3, respectively. Conclusions MALDI-TOF MS combined with BioExplorer software, when used as a new method to study the E. faecium bloodstream infection, may find the differential polypeptides between infection group and control group, and can effectively establish a diagnostic model of bloodstream infection. β2-microglobulin and complement C3 are expected to become the new biomarkers for auxiliary diagnosis of bacterial bloodstream infection.

Distribution and transmission characteristics of thirty-five dual genotype vancomycin-resistant Enterococcus faecium carrying both vanA and vanM in an intensive care unit / 中华传染病杂志

Objective To investigate the distribution and transmission characteristics of vancomycin -resistant Enterococcus faecium (VREF) carrying both vanA and vanM in the intensive care unit.Methods VREF strains were isolated from patients in the intensive care unit of Jinshan Hospital , Fudan University in Shanghai from 2013 to 2017.Antimicrobial susceptibilities of the VREF strains to nine antibiotics , including vancomycin, teicoplanin, linezolid and chloromycetin , were tested by broth microdilution method.Multiple polymerase chain reaction (PCR) was used for van genotyping and pulsed field gel electrophoresis (PFGE) was used for homology analysis.Results Thirty-five strains were mainly isolated from urine (16 strains), blood (11 strains), feces ( five strains ), bile ( two strains ) and pleural effusion ( one strain ).All the strains (100.00%) were resistant to vancomycin , ampicillin and levofloxacin , but only 40.00% were resistant to teicoplanin.All the strains were sensitive to linezolid.The results of van genotyping showed that 33 (94.3%) strains belonged to vanA and vanM dual genotype VREF, and the other two were vanA type VREF.PFGE results showed that 35 strains could be divided into 14 PFGE patterns, and seven out of 10 strains isolated in 2014 were identical and the other three belonged to three different PFGE patterns.Conclusions A dual genotype VREF carrying both vanA and vanM has been emerging and spreading in the intensive care unit of Jinshan Hospital , Fudan University in Shanghai.

Distribución de especies y susceptibilidad antimicrobiana en cepas de Enterococcus de origen clínico / Distribution of species and antimicrobial susceptibility in Enterococcus strains of clinical origin

A fin de establecer la frecuencia de aislamiento de las diferentes especies de enterococos, su distribución de acuerdo al tipo de muestra y servicio de atención al paciente y determinar la resistencia antimicrobiana, se analizaron 1.624 cepas provenientes de cultivos bacteriológicos de pacientes atendidos en el Centro de Referencia Bacteriológica del Servicio Autónomo Hospital Universitario de Maracaibo, durante el período Enero 2010 - Diciembre 2015. Las especies más frecuentes fueron E. faecalis (52,46%) y E. faecium (41,38%). El mayor número de cepas se obtuvo a partir de muestras de piel y tejidos blandos (54,92%), orina (23,15%) y sangre (17,27%). Los servicios con mayor frecuencia de aislamiento fueron: hospitalización de adultos (47,71%) y hospitalización pediátrica (16,38%). No se encontró asociación estadísticamente significativa entre la especie de enterococos y el tipo de muestra o el servicio de atención al paciente (p>0,05). Se detectó más resistencia en E. faecium que en E. faecalis. Los enterococos están adquiriendo cada vez mayor resistencia antimicrobiana y, por lo tanto, se hace necesario mantener una vigilancia permanente sobre ellos, realizar su adecuada identificación y detectar oportunamente la resistencia, con el fin de aplicar medidas preventivas adecuadas antes de que estos microorganismos causen un mayor impacto intrahospitalario.

In order to establish the frequency of isolation of the different species of enterococci, their distribution according to the type of sample and patient care service and determine the antimicrobial resistance, 1,624 strains obtained from bacteriological cultures of patients attended in the Bacteriological Reference Center at the Autonomous Service University Hospital of Maracaibo, during the period January 2010 - December 2015, were analyzed. The most frequent species were E. faecalis (52.46%) and E. faecium (41.38%). The greatest number of strains was obtained from skin and soft tissues samples (54.92%), urine (23.15%) and blood (17.27%). Services with increased frequency of isolation were: hospitalization of adults (47.71%) and pediatric hospitalization (16.38%). It did not find statistically significant association between the specie of enterococci and sample type, or patient care service (p > 0.05). It was detected more resistance in E. faecium than in E. faecalis. The enterococci are acquiring ever greater antimicrobial resistance, and therefore, it is necessary to maintain permanent vigilance over them, perform their proper identification and timely detect resistance, in order to apply preventive measures before these microorganisms cause a greater intrahospital impact.

van genotype and molecular typing of clinical isolates of vancomycin-resistant Enterococcus / 中国感染与化疗杂志

Objective To determine the genotype and molecular typing of vancomycin-resistant Enterococcus (VRE).Methods Seventeen clinical isolates of VRE were collected in 2016.The strains were identified to species and confirmed by 16S rRNA sequencing.The minimum inhibitory concentration of antimicrobial agents was determined by microdilution method and agar dilution method.Multilocus sequence typing (MLST) was used for molecular typing.Results The VRE strains were confirmed as Enterococcusfaecium by 16S rRNA sequencing.All strains were resistant to vancomycin,but only 12 strains were resistant to teicoplanin.The vanA gene was identified in 13 of the 17 strains.The vanM gene was detected in 9 strains.Both vanA and vanM genes were identified in five of the 17 strains.Six MLST types were identified in the 17 strains,including ST78 (n=8),ST80 (n=4),ST555 (n=2),and one each for ST117,ST262 and ST341.Conclusions The van genotype was primarily vanA (76.5％) and vanM (52.9％) in clinical isolates of VRE.The VRE strains carrying both vanA and vanM were found for the first time.

Formação de biofilmes monotípicos e heterotípicos por Enterococcus faecalis E Enterococcus faecium em dentina radicular / Formation of monotypic and heterotypic biofilm by Enterococcus faecalis E Enterococcus faecium in root dentin

E. faecalis e E. faecium possuem grande relevância nas infecções hospitalares por apresentarem facilidade em adquirir resistência aos antibióticos. E. faecalis também apresentam alta prevalência nas infecções endodônticas, entretanto a importância de E. faecium para a odontologia ainda precisa ser esclarecida. Assim, o objetivo desse estudo foi comparar cepas clínicas de E. faecium com as cepas de E. faecalis em relação a capacidade de formação de biofilme na dentina radicular e penetração nos túbulos dentinários. Além disso, foi avaliada a interação dessas espécies em biofilmes mistos. Para a realização desse estudo, foram utilizadas cepas clínicas, previamante, isoladas de canais radiculares com infecções endodõnticas e identificadas pelo PCR multiplex. Entre as cepas isoladas, foram selecionadas 4 cepas de E. faecalis e 2 cepas de E. faecium. Primeiramente, foi realizado a formação dos biofilmes monotípicos das cepas de E. faecalis e E. faecium sobre dentinas radiculares de dentes bovinos. Os biofilmes foram formados em placas de microtitulação por diferentes tempos: 2, 4, 6, 24, 48, 72, 96 e 120 horas. Os biofilmes formados foram, então, analisados pela contagem de células viáveis (UFC/mL) e quantificação da biomassa total (método do cristal violeta). Além disso, os biofilmes foram analisados por Microscopia Eletrônica de Varredura (MEV) procurando-se observar a penetração das células de E. faecalis e E. faecium nos túbulos dentinários. A seguir foram formados biofilmes heterotípicos de E. faecalis e E. faecium para estudo das interações ecológicas estabelecidas entre as espécies. A análise dos biofilmes heterotípicos foi feita pela quantificação da biomassa total (cristal violeta) procurando-se detectar a presença de relações sinérgicas ou antagônicas. Os resultados foram submetidos à Análise de Variância (ANOVA) e teste de Tukey, considerando-se nível de 5%. Os resultados obtidos na contagem de UFC/mL dos biofilmes monotípicos, revelaram que as 6 cepas testadas apresentam grande capacidade para formar biofilmes na dentina radicular, alcançando valores de UFC/mL entre 8 a 12 log de acordo com o tempo de observação. Em relação a análise das imagens de MEV, as cepas clínicas de E. faecalis e E. faecium demonstraram capacidade semelhante para formar biofilmes e penetrar nos túbulos dentinários. Na comparação da quantificação da biomassa dos biofilmes monotípicos e heterotípicos, observamos que a interação das cepas clínicas E. faecalis e E. faecium favoreceu a adesão e crescimento do biofilme. Assim, concluiuse que as cepas de E. faecalis e E. faecium apresentam a mesma capacidade de formar biofilmes sobre a superfície radicular. Além disso, em biofilmes mistos, essas duas espécies estabelecem relações ecológicas sinérgicas, aumentando significativamente a formação de biofilmes(AU)

E. faecalis and E. faecium have a high relevance in hospital infections because they are easy to acquire resistance to antibiotics. E. faecalis also present high prevalence in endodontic infections, however the importance of E. faecium for dentistry still needs to be clarified. Thus, the objective of this study was to compare clinical strains of E. faecium with strains of E. faecalis in relation to the capacity of biofilm formation in root dentin and penetration into the dentin tubules. In addition, the interaction of these species in mixed biofilms was evaluated. In order to perform this study, clinical strains were used, pre-determined, isolated from root canals with endodontic infections and identified by multiplex PCR. Among the isolated strains, 4 strains of E. faecalis and 2 strains of E. faecium were selected. Firstly, the formation of the monotypic biofilms of the strains of E. faecalis and E. faecium on root dentin of bovine teeth was carried out. The biofilms were formed in microtiter plates at different times: 2, 4, 6, 24, 48, 72, 96 and 120 hours. The biofilms formed were then analyzed by counting viable cells (CFU / mL) and quantification of total biomass (violet crystal method). In addition, the biofilms were analyzed by Scanning Electron Microscopy (SEM), aiming to observe the penetration of E. faecalis and E. faecium cells into the dentin tubules. Then, heterophilic biofilms of E. faecalis and E. faecium were formed to study the ecological interactions established between the species. The analysis of the heterotypic biofilms was made by quantifying the total biomass (violet crystal) in order to detect the presence of synergistic or antagonistic relationships The results were submitted to Analysis of Variance (ANOVA) and Tukey test, considering a level of 5%. The results obtained in the CFU / mL count of the monotypic biofilms revealed that the six strains tested had a great capacity to form biofilms in the root dentin, reaching values of CFU / mL between 8 and 12 log according to the time of observation. In relation to SEM images, the clinical strains of E. faecalis and E. faecium demonstrated similar capacity to form biofilms and to penetrate the dentinal tubules. In the comparison of the biomass quantification of the monotypic and heterotypic biofilms, we observed that the interaction of the clinical strains E. faecalis and E. faecium favored the adhesion and growth of the biofilm. Thus, it was concluded that strains of E. faecalis and E. faecium have the same ability to form biofilms on the root surface. In addition, in mixed biofilms, these two species establish synergistic ecological relationships, significantly increasing the formation of biofilms(AU)

Clonal dissemination of vancomycin-resistant Enterococcus faecium ST412 in a Brazilian region

ABSTRACT Vancomycin-resistant Enterococcus faecium (VREfm) has emerged as an important global nosocomial pathogen, and this trend is associated with the spread of high-risk clones. Here, we determined the genetic and phenotypic features of 93 VREfm isolates that were obtained from patients in 13 hospitals in Vitória, Espírito Santo, Brazil, during 2012-2013. All the isolates were vancomycin-resistant and harbored the vanA gene. Only 6 (6.5%) of the VREfm isolates showed the ability to form biofilm. The 93 isolates analyzed belong to a single pulsed-field gel electrophoresis lineage and presented six subtypes. MLST genotyping showed that all VREfm belonged to ST412 (the high-risk clone, hospital-adapted). The present study describes the dissemination of ST412 clone in the local hospitals. The clonal spread of these ST412 isolates in the area we analyzed as well as other hospitals in southeastern Brazil supports the importance of identifying and controlling the presence of these microorganisms in health care-related services.

The clinical effect of live Bacillus Subtilis and Enterococcus Faecium granules combined with blue light irradiation in the treatment of neonatal jaundice / 中国基层医药

Objective To explore the clinical effect of live Bacillus Subtilis and Enterococcus Faecium granulescombined with blue light irradiation in the treatment of neonatal jaundice.Methods 90 patients with neonatal jaundice were randomly divided into control group and observation group.The control group received the blue light treatment,the observation group was given blue light irradiation combined with live Bacillus Subtilis and Enterococcus Faecium granules treatment,the patients were treated for 7 days.The clinical efficacy,serum bilirubin and main indicators of complex constant time were observed in the two groups.Results After treatment,the total effective rate of the observation group was 95.6％,which of the control group was 68.9％,the difference was statistically significant (x2 =5.473,P ＜ O.05).After treatment for 1 day,3 days,7 days,the serum bilirubin values of the observation group were (185.6 ± 25.1) μmol/L,(136.7 ± 19.1) μmol/L,(82.1 ± 10.3) μmol/L,respectively,which of the control group were (205.3 ± 26.4) μmol/L,(184.1 ± 20.3) μmol/L,(128.4 ± 16.7) μmol/L,the differences between the two groups were statistically significant (t =3.628,11.408,15.829,all P ＜ 0.05).The recovery time of serum bilirubin in the observation group was (7.1 ± 2.4) d,which of the control group was (12.9 ±3.1) d,there was significant difference between the two groups (t =9.924,P ＜ 0.05).The hospitalization time of observation group was (3.5 ± 1.1) d,which of the control group was (5.8 ± 1.5) d,the difference between the two groups was statistically significant (t =8.295,P ＜ 0.05).Conclusion Live Bacillus Subtilis and Enterococcus Faecium granules combined with blue light irradiation in the treatment of neonatal jaundice is helpful to improve the clinical efficacy,reduce the treatment time and improve the clinical symptoms.

Distribution characteristics and diversity of virulence genes in clinical isolates of vancomycin-resistant enterococci / 中国人兽共患病学报

The aim of this study is to determine the diversity of virulence genes carried by different vancomycin-resistant enterococci (VRE),which will provides a basis for studying pathogenic mechanism of VRE.Microdilution-based drug sensitivity test was applied to detect the vancomycin resistance of 490 Enterococcus faecium isolates and 862 Enterococcus faecalis isolates in Zhejiang area.The seven virulence genes (ace,asa1,cylA,efaA,esp,gelE and hyl) in the isolates of VRE were detected by PCR.According to the results of drug sensitivity test,10％ of the E.faecium isolates (49/490) and 0.8％ of the E.faecalis (7/862) were identified as VRE.In the vancomycin-resistant E.faecium isolates,five isolates were negative for any of the target genes and the other 44 isolates were positive for asa1,esp,gelE and hyl genes alone,in which the esp (73.5％,36/49) and hyl (53.1％,26/49) were the predominant genes and single or double virulence genes acted as the major carrying models.Except for the hyl gene,the vancomycin-resistant E.faecalis isolates were positive for the other six pathogenic genes,and the isolates could carry 3-6 pathogenic genes.All the data indicate that E.faeciurn is the major species of VRE in the local area,and the carrying rate,types and models of virulence genes in the vancomycin-resistant E.faecium and E.faecalis isolates are obviously different.

Surveillance of antibiotic resistance of Enterococcus faecalis and Enterococcus faecium in Chongqing in 2015 / 国际检验医学杂志

Objective To analyze drug resistance data of Enterococcus faecalis and Enterococcus faecium,submitted by member units of Chongqing bacterial drug resistance monitoring network,and to provide the basis for our city effective application of antimicrobial agents and the reference.Methods Target bacteria identification and drug susceptibility test were performed by member units,according to the national technology scheme of bacterial drug resistance monitoring network and the results were determined according to standards published by Clinic and Laboratory Standard Institute(CLSI) in 2015.WHONET5.6 software was used to analyze drug susceptibility,and drug resistance difference was analyzed by using SPSS21.0 software.Results A total of un-repeated 1 811 strains of Enterococcus faecalis and 1 601 strains of Enterococcus faecium,accounting for 13.1% of all positive strains.The resistant rates of the two kinds of bacteria to vancomycin were 0.5% and 1.8%,to rinathiazole amine were 2.5% and 0.5% respectively.Tigecycline resistant strains were not founded.The resistant rate of Enterococcus feaclis to ouinupristin/dalfopristin was 90.1%,to tetracycline was 78.8%,to high concentration of gentamicin was 43.0%,to penicillin,ampicillin and nitrofurantoin was less than 7%.Except ouinupristin/dalfopristin and tetracycline,the resistant rate of Enterococcus faecium to the other drugs were significantly higher than Enterococcus faecalis(P<0.05).Strains isolated from children and adult patients,Intensive Care Unit(ICU) and un-ICU patients were with differences of drug resistance(P<0.05).Conclusion Most of Enterococcus infection could be caused by Enterococcus faecalis and Enterococcus faecium.Monitoring of drug resistance might be helpful for rational and effective usage of antimicrobial agents.

Observation of clinical curative effect of Live Combined Bacillus Subtilis and Enterococcus Faecium Enteric-coated Capsules combined with trimebutine maleate and psychological intervention in the treatment of functional diarrhea / 中国生化药物杂志

Objective To study the clinical efficacy of live combined bacillus subtilis and enterococcus faecium enteric-coated capsules combined with trimebutine maleate and psychological intervention in the treatment of functional diarrhea. Methods 100 patients with functional diarrhea treated in our hospital from May 2015 to August 2016 were selected and randomly divided into the control group and the experimental group, with 50 patients in each group. The control group were treated with trimebutine maleate treatment, the experimental group were treated with live combined bacillus subtilis and enterococcus faecium enteric-coated capsules trimebutine therapy and psychological intervention, pay attention to the psychological status of patients and patients, strengthen communication and exchanges, increase confidence in the treatment and the treatment compliance of patients. The therapeutic effects of the experimental group and the control group were compared and analyzed. Results After the corresponding treatment, the effective rate was 92.0％ in the experimental group and 84.0％ in the control group. The difference was statistically significant (P<0.05). There was no obvious adverse reaction in the experimental group and the control group. After treatment, the number of diarrhea in the experimental group was (3.0±0.3), significantly less than that of the control group (4.3±0.3), the difference was statistically significant (P<0.05). Conclusion The clinical effect of live combined bacillus subtilis and enterococcus faecium enteric-coated capsules combined with trimebutine maleate and psychological intervention in the treatment of functional diarrhea is good, can improve clinical symptoms in a large extent, has the meaning of promotion.