Significant production of vanillin and in vitro amplification of ech gene in local bacterial isolates / Produção significativa de vanilina e amplificação in vitro do gene ech em isolados bacterianos locais

Abstract Vanillin is the major component which is responsible for flavor and aroma of vanilla extract and is produced by 3 ways: natural extraction from vanilla plant, chemical synthesis and from microbial transformation. Current research was aimed to study bacterial production of vanillin from native natural sources including sewage and soil from industrial areas. The main objective was vanillin bio-production by isolating bacteria from these native sources. Also to adapt methodologies to improve vanillin production by optimized fermentation media and growth conditions. 47 soil and 13 sewage samples were collected from different industrial regions of Lahore, Gujranwala, Faisalabad and Kasur. 67.7% bacterial isolates produced vanillin and 32.3% were non-producers. From these 279 producers, 4 bacterial isolates selected as significant producers were; A3, A4, A7 and A10. These isolates were identified by ribotyping as A3 Pseudomonas fluorescence (KF408302), A4 Enterococcus faecium (KT356807), A7 Alcaligenes faecalis (MW422815) and A10 Bacillus subtilis (KT962919). Vanillin producers were further tested for improved production of vanillin and were grown in different fermentation media under optimized growth conditions for enhanced production of vanillin. The fermentation media (FM) were; clove oil based, rice bran waste (residues oil) based, wheat bran based and modified isoeugenol based. In FM5, FM21, FM22, FM23, FM24, FM30, FM31, FM32, FM34, FM35, FM36, and FM37, the selected 4 bacterial strains produced significant amounts of vanillin. A10 B. subtilis produced maximum amount of vanillin. This strain produced 17.3 g/L vanillin in FM36. Cost of this fermentation medium 36 was 131.5 rupees/L. This fermentation medium was modified isoeugenol based medium with 1% of isoeugenol and 2.5 g/L soybean meal. ech gene was amplified in A3 P. fluorescence using ech specific primers. As vanillin use as flavor has increased tremendously, the bioproduction of vanillin must be focused.

Resumo A vanilina é o principal componente responsável pelo sabor e aroma do extrato de baunilha e é produzida de três formas: extração natural da planta da baunilha, síntese química e transformação microbiana. A pesquisa atual teve como objetivo estudar a produção bacteriana de vanilina a partir de fontes naturais nativas, incluindo esgoto e solo de áreas industriais. O objetivo principal era a bioprodução de vanilina por meio do isolamento de bactérias dessas fontes nativas. Também para adaptar metodologias para melhorar a produção de vanilina por meio de fermentação otimizada e condições de crescimento. Foram coletadas 47 amostras de solo e 13 de esgoto de diferentes regiões industriais de Lahore, Gujranwala, Faisalabad e Kasur; 67,7% dos isolados bacterianos produziram vanilina e 32,3% eram não produtores. Desses 279 produtores, 4 isolados bacterianos selecionados como produtores significativos foram: A3, A4, A7 e A10. Esses isolados foram identificados por ribotipagem como fluorescência A3 Pseudomonas (KF408302), A4 Enterococcus faecium (KT356807), A7 Alcaligenes faecalis (MW422815) e A10 Bacillus subtilis (KT962919). Os produtores de vanilina foram posteriormente testados para produção aprimorada de vanilina e foram cultivados em diferentes meios de fermentação sob condições de crescimento otimizadas para produção aprimorada de vanilina. Os meios de fermentação (FM) foram: à base de óleo de cravo, à base de resíduos de farelo de arroz (resíduos de óleo), à base de farelo de trigo e à base de isoeugenol modificado. Em FM5, FM21, FM22, FM23, FM24, FM30, FM31, FM32, FM34, FM35, FM36 e FM37, as 4 cepas bacterianas selecionadas produziram quantidades significativas de vanilina. A10 B. subtilis produziu quantidade máxima de vanilina. Essa cepa produziu 17,3 g / L de vanilina em FM36. O custo desse meio de fermentação 36 foi de 131,5 rúpias / L. Esse meio de fermentação foi um meio à base de isoeugenol modificado com 1% de isoeugenol e 2,5 g / L de farelo de soja. O gene ech foi amplificado em A3 P. fluorescence usando primers específicos para ech. Como o uso da vanilina como sabor aumentou tremendamente, a bioprodução da vanilina deve ser focada.

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.

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

Hallazgos microbiológicos y susceptibilidad antimicrobiana en cirugía abdominal de urgencia / Microbiological findings and antimicrobial susceptibility in emergency abdominal surgery

Resumen Introducción: Es importante mantener programas de vigilancia bacteriana para disminuir resistencia y definir esquemas farmacológicos adecuados. Los pacientes con abdomen agudo representan un grupo microbiológico especial. Objetivos: Hacer una revisión de agentes patógenos en pacientes adultos operados en nuestro Servicio de Urgencia por patología abdominal con líquido libre y analizar los resultados obtenidos de cultivos respecto a las cepas y la susceptibilidad a los antibióticos. Materiales y Método: Estudio de cohorte prospectiva con estadística descriptiva. Se incluyen pacientes consecutivos, mayores de 18 años, operados por abdomen agudo que presentan líquido libre intraperitoneal entre noviembre de 2017 y abril de 2018. Se excluyen casos con terapia antimicrobiana, hospitalización y/o cirugía en los 3 meses previos. Se registran los cultivos positivos, cepas aisladas, susceptibilidad antimicrobiana, datos demográficos y evolución clínica. Resultados: De 63 pacientes 55% fueron hombres, edad promedio 52,2 años. Las patologías más frecuentes fueron de origen apendicular (62%) y de causa entérica (30%). En un 44% el cultivo fue positivo y en 36% con más de un germen. Escherichia coli fue el patógeno más frecuente (64,2%) seguidos de Enterococcus faecium y Streptococcus anginosus (7,1%). De los otros patógenos cultivados sólo se observó resistencia múltiple en un caso aislado de Morganella Morganii. Conclusiones: Estos datos constituyen la realidad microbiológica local en abdomen agudo. La Escherichia Coli sigue siendo el germen más frecuente, debe enfrentarse con profilaxis y tratamiento antibiótico adecuado. Es necesario mantener vigilancia microbiología local para un manejo acorde.

Introduction: It is important to maintain bacterial surveillance programs to decrease resistance and define adequate pharmacological schemes. Patients with abdomen represent a special microbiological group. Objetives: Make a review of pathogens in adult patients operated in our Emergency Service for abdominal pathology with free fluid and analyze the results obtained from cultures with respect to the strains and susceptibility to antibiotics. Materials and Method: Prospective cohort study with descriptive statistics. We include consecutive patients, older than 18 years old, operated on by abdomen who present free intraperitoneal fluid between November 2017 and April 2018. Cases with antimicrobial therapy, hospitalization and/or surgery 3 months prior are excluded. Positive cultures, isolated strains, antimicrobial susceptibility, demographic data and clinical evolution are recorded. Results: Of 63 patients, 55% were men and the average age was 52.2 years. The most frequent pathologies were of appendicular origin (62%) and of enteric origin (30%). In 44% the crop was positive and in 36% with more than one germ. Escherichia coli was the most frequent pathogen (64.2%) followed by Enterococcus faecium and Streptococcus anginosus (7.1%). Of the others, cultivated pathogens have only observed multiple resistance in an isolated case of Morganella Morganii. Conclusions: These data include the local microbiological reality in acute abdomen. Escherichia coli is still the most frequent germ that must be faced with the profile and the appropriate treatment. It is necessary to maintain local microbiology surveillance for a proper management.

Antimicrobial resistance and antibiotic consumption in Mexican hospitals / Resistencia antimicrobiana y consumo de antibióticos en hospitales mexicanos

Abstract: Objective: To establish the current situation of antimicrobial resistance and antibiotic consumption in Mexican hospitals. Materials and methods: Antimicrobial susceptibility data from blood and urine isolates were collected. Defined daily dose (DDD) of antibiotic consumption/100 occupied beds (OBD) was calculated. Results: Study period: 2016 and 2017. Of 4 382 blood isolates, E. coli and K. pneumoniae were most frequently reported, with antimicrobial resistance >30% for most drugs tested, only for carbapenems and amikacin resistance were <20%. A. baumannii had antimicrobial resistance >20% to all drugs. Resistance to oxacillin in S. aureus was 20%. From 12 151 urine isolates, 90% corresponded to E. coli; resistance to ciprofloxacin, cephalosporins and trimethoprim/sulfamethoxazole was >50%, with good susceptibility to nitrofurantoin, amikacin and carbapenems. Global median antimicrobial consumption was 57.2 DDD/100 OB. Conclusions: This report shows a high antimicrobial resistance level in Gram-negative bacilli and provides an insight into the seriousness of the problem of antibiotic consumption.

Resumen: Objetivo: Establecer la situación actual de la resistencia antimicrobiana y el consumo de antibióticos en hospitales mexicanos. Material y métodos:F Se colectaron datos de susceptibilidad antimicrobiana de aislamientos de sangre y orina. Se calculó la dosis diaria definida (DDD) del consumo de antibióticos/100 estancias. Resultados: Periodo de estudio de 2016 a 2017. De 4 382 aislamientos en sangre, E. coli y K. pneumoniae fueron las más frecuentes, con resistencia >30% a la mayoría de las drogas evaluadas; sólo para carbapenémicos y amikacina la resistencia fue <20%. A. baumannii tuvo resistencia >20% a todos los fármacos. La resistencia a oxacilina en S. aureus fue de 20%. De 12 151 aislamientos en urocultivos, 90% correspondió a E. coli; la resistencia a ciprofloxacina, cefalosporinas y trimetoprima/sulfametoxazol fue >50%, con buena susceptibilidad a nitrofurantoína, amikacina y carbapenémicos. La mediana del consumo global de antibióticos en DDD/100 estancias fue de 57.2. Conclusiones: Este reporte muestra el nivel elevado de resistencia en bacilos Gram-negativos y brinda una perspectiva de la gravedad del problema del consumo de antibióticos.

Antibacterial Activity of Sophorolipids from Candida bombicola Against Human Pathogens

Abstract Sophorolipids are glycolipids that have natural antimicrobial properties and present great potential in the pharmaceutical field. The present study aimed to produce sophorolipids from Candida bombicola using a chicken fat-based medium and evaluate the antimicrobial activity against Gram-negative (Proteus mirabilis, Escherichia coli, Salmonella enterica subsp. enterica) and Gram-positive bacteria (Enterococcus faecium, Staphylococcus aureus and Streptococcus mutans). The production of sophorolipids reached 27.86 g L-1. Based on the structural characterization, 73.55% of the sophorolipids present a mixture of acidic monoacetylated C18:2 and lactonic diacetylated C16:0, and 26.45% were present in the diacetylated C18:1 lactonic form. Bacteria submitted to sophorolipid exposure showed a reduction in viability at doses of 500 μg mL-1 and 2,000 μg mL-1 against Gram-positive and Gram-negative bacteria, respectively. These results suggest that sophorolipids produced in chicken fat medium may be used as antimicrobial agents to prevent or eliminate contamination by different pathogens.

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.

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.

Antimicrobial Resistance in Bacterial Isolates Recovered from Nursing Hospitals between 2014 and 2017 / 대한임상미생물학회지

BACKGROUND: Antimicrobial resistance (AMR) is an issue not only with regard to public health, but also in terms of economic impact. AMR surveillance has mainly been carried out in general hospitals, and not in nursing hospitals. This study was conducted to investigate the AMR rate for bacterial strains isolated from nursing hospital samples.METHODS: Antimicrobial susceptibility testing (AST) results from a total of 23,518 bacterial isolates recovered from clinical specimens taken in 61 nursing hosals were analyzed. AST was conducted using Vitek 2 with AST cards specific for the bacterial strains.RESULTS: A total of 19,357 Gram-negative and 4,161 Gram-positive bacterial strains were isolated. Pseudomonas aeruginosa (n=6,384) and Escherichia coli (n=5,468) were the most prevalent bacterial species and, among Gram-positive bacteria, Staphylococcus aureus (n=1,565) was common. The AMR rate was high for the following strains: cefotaxime-resistant Klebsiella pneumoniae, 77.4%; cefotaxime-resistant E. coli, 70.6%; imipenem-resistant Acinetobacter baumannii, 90.3%; imipenem-resistant P. aeruginosa, 49.3%; oxacillin-resistant S. aureus, 81.1%, penicillin-resistant Enterococcus faecalis, 44.8%, and vancomycin-resistant Enterococcus faecium, 53.5%. AMR rate change varied by bacterial species and antimicrobial drug.CONCLUSION: AMR rates of major pathogens from nursing hospitals were higher than those from general hospitals with the exception of imipenem-resistant A. baumannii. Continuous monitoring and infection control strategies are needed.

The antimicrobial activity of ephedrine and admixture of ephedrine and propofol: an in vitro study / A atividade antimicrobiana de efedrina e da combinação de efedrina e propofol: um estudo in vitro

Abstract Introduction Propofol and Ephedrine are commonly used during anesthesia maintenance, the former as a hypnotic agent and the later as a vasopressor. The addition of propofol to ephedrine or administration of ephedrine before propofol injection is useful for decreasing or preventing propofol related hemodynamic changes and vascular pain. This in vitro study evaluated the antibacterial effect on common hospital-acquired infection pathogens of ephedrine alone or combined with propofol. Material and method The study was performed in two stages. In the first, the Minimum Inhibitory Concentration of propofol and ephedrine alone and combined was calculated for Escherichia coli, Enterococcus faecium, Staphylococcus aureus, Pseudomonas aeruginosa, and a clinical isolate of Acinetobacter spp. at 0, 6, 12 and 24 h, using the microdilution method. In the second stage, the same drugs and combination were used to determine their effect on bacterial growth. Bacterial solutions were prepared at 0.5 MacFarland in sterile 0.9% physiological saline and diluted at 1/100 concentration. Colony numbers were measured as colony forming units.mL-1 at 0, 2, 4, 6, 8, 10 and 12th hours. Results Ephedrine either alone or combined with propofol did not have an antimicrobial effect on Escherichia coli, Enterococcus faecium, or Pseudomonas aeruginosa and this was similar to propofol. However, ephedrine alone and combined with propofol was found to have an antimicrobial effect on Staphylococcus aureus and Acinetobacter species at 512 mcg.mL-1 concentration and significantly decreased bacterial growth rate. Conclusion Ephedrine has an antimicrobial activity on Staphylococcus aureus and Acinetobacter species which were frequently encountered pathogens as a cause of nosocomial infections.

Resumo Introdução Propofol e efedrina são fármacos comumente usados durante a manutenção da anestesia, o primeiro como agente hipnótico e o segundo como vasopressor. A adição de propofol à efedrina ou a administração de efedrina antes da injeção de propofol é útil para diminuir ou prevenir alterações hemodinâmicas e dor vascular relacionadas ao propofol. Este estudo in vitro avaliou o efeito antibacteriano de efedrina, isolada ou em combinação com propofol, em patógenos comuns implicados em infecção hospitalar. Material e método O estudo foi feito em duas etapas. Na primeira, a concentração inibitória mínima (CIM) de propofol e de efedrina isolada e em combinação foi calculada para Escherichia coli, Enterococcus faecium, Staphylococcus aureus, Pseudomonas aeruginosa e um isolado clínico de Acinetobacter spp às 0, 6, 12 e 24 horas, com o método de microdiluição. Na segunda etapa, o mesmo fármaco e sua combinação foram usados para determinar seus efeitos no crescimento bacteriano. As soluções bacterianas foram preparadas em soro fisiológico a 0,9% em 0,5 McFarland e diluídas a uma concentração de 1/100. Os números das colônias foram medidos como ufc.mL-1 às 0, 2, 4, 6, 8, 10 e 12 horas. Resultados Efedrina isolada ou em combinação com propofol não apresentou efeito antimicrobiano sobre E. coli, E. faecium ou P. aeruginosa, um resultado semelhante ao de propofol. Porém, efedrina isolada e em combinação com propofol apresentou efeito antimicrobiano sobre Staphylococcus aureus e Acinetobacter spp, em concentração de 512 mcg.mL-1, e redução significativa da taxa de crescimento bacteriano. Conclusão Efedrina tem atividade antimicrobiana em S. aureus e Acinetobacter spp, patógenos frequentemente identificados como causa de infecções nosocomiais.

Inhibitory Effect of Metal Surface on the Antimicrobial Resistance Microorganism / 대한임상미생물학회지

BACKGROUND: The aim of this study was to comparatively evaluate the bactericidal effects of copper, brass (copper 78%, tin 22%), and stainless steel against methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus faecium (VREFM), and multidrug-resistant Pseudomonas aeruginosa (MRPA). METHODS: The isolates (MRSA, VREFM, MRPA) used in this study were mixed wild type 3 strains isolated from patients treated at Uijeongbu St. Mary's Hospital in 2017. These strains showed patterns of multidrug resistance. The lyophilized strains were inoculated into and incubated for 24 hr in tryptic soy broth at 35℃. The initial bacterial inoculum concentration was adjusted to 105 CFU/mL. A 100-mL bacterial suspension was incubated in containers made of brass (copper 78%, tin 22%), copper (above 99% purity), and stainless steel at 35℃. Viable counts of bacteria strains were measured for 9 days. RESULTS: In this study, the bactericidal effects of copper and brass on MRSA, VREFM, and MRPA were verified. The bactericidal effect of stainless steel was much weaker than those of copper and brass. The bactericidal effect was stronger on MRPA than on MRSA or VREFM. CONCLUSION: To prevent cross infection of multidrug resistant bacteria in hospitals, further studies of longer duration are needed for testing of copper materials on objects such as door knobs, faucets, and bed rails.

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)

Comparison of Enterococcus faecium Bacteremic Isolates from Hematologic and Non-hematologic Patients: Differences in Antimicrobial Resistance and Molecular Characteristics

BACKGROUND: Enterococcus faecium, especially vancomycin-resistant E. faecium (VREfm), is a major concern for patients with hematologic diseases. Exposure to antibiotics including fluoroquinolone, which is used as a routine prophylaxis for patients with hematologic (MH) diseases, has been reported to be a risk factor for infection with vancomycin-resistant eneterocci. We compared the characteristics of E. faecium isolates according to their vancomycin susceptibility and patient group (MH vs non-MH patients). METHODS: A total of 120 E. faecium bacteremic isolates (84 from MH and 36 from non-MH patients) were collected consecutively, and their characteristics (susceptibility, multilocus sequence type [MLST], Tn1546 type, and the presence of virulence genes and plasmids) were determined. RESULTS: Among the vancomycin-susceptible E. faecium (VSEfm) isolates, resistance to ampicillin (97.6% vs 61.1%) and high-level gentamicin (71.4% vs 38.9%) was significantly higher in isolates from MH patients than in those from non-MH patients. Notably, hyl, esp, and pEF1071 were present only in isolates with ampicillin resistance. Among the VREfm isolates, ST230 (33.3%) and ST17 (26.2%) were predominant in MH patients, while ST17 (61.1%) was predominant in non-MH patients. Plasmid pLG1 was more prevalent in E. faecium isolates from MH patients than in those from non-MH patients, regardless of vancomycin resistance. Transposon analysis revealed five types across all VREfm isolates. CONCLUSIONS: The antimicrobial resistance profiles and molecular characteristics of E. faecium isolates differed according to the underlying diseases of patients within the same hospital. We hypothesize that the prophylactic use of fluoroquinolone might have an effect on these differences.

Clinical features of Enterococcus faecium meningitis in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To summarize the clinical features of Enterococcus faecium meningitis in children.</p><p><b>METHODS</b>The clinical data of nine children with Enterococcus faecium meningitis were analyzed.</p><p><b>RESULTS</b>In all the nine children, Enterococcus faecium was isolated from blood, cerebrospinal fluid, or peripherally inserted central catheters; 6 (67%) patients were neonates, 2 (22%) patients were younger than 6 months, and 1 (11%) patient was three years and four months of age. In those patients, 56% had high-risk factors before onset, which included intestinal infection, resettlement of drainage tube after surgery for hydrocephalus, skull fracture, perinatal maternal infection history, and catheter-related infection. The main symptoms were fever and poor response. In those patients, 22% had seizures; no child had meningeal irritation sign or disturbance of consciousness. The white blood cell count and level of C-reactive protein were normal or increased; the nucleated cell count in cerebrospinal fluid was normal or mildly elevated; the protein level was substantially elevated; the glucose level was decreased. The drug sensitivity test showed that bacteria were all sensitive to vancomycin and the vancomycin treatment was effective. Only one child had the complication of hydrocephalus.</p><p><b>CONCLUSIONS</b>Enterococcus faecium meningitis occurs mainly in neonates and infants. The patients have atypical clinical features. A high proportion of patients with Enterococcus faecium meningitis have high-risk factors. Enterococcus faecium is sensitive to vancomycin.</p>

Gliotoxin is Antibacterial to Drug-resistant Piscine Pathogens

By activity-guided fractionation, gliotoxin was isolated as an antibacterial metabolite of the fungus Penicillium decumbens which was derived from the jellyfish Nemopilema nomurai. Gliotoxin was further evaluated for antibacterial activity against several piscine and human MDR (multidrug resistance) pathogens. Gliotoxin showed significant antibacterial activity against Gram-positive piscine pathogens such as Streptococcus iniae FP5228, Streptococcus iniae FP3187, Streptococcus parauberis FP3287, Streptococcus parauberis SPOF3K, S. parauberis KSP28, and Lactococcus garvieae FP5245. Gliotoxin showed strong activity especially against S. parauberis SPOF3K and S. iniae FP5228, which are resistant to oxytetracycline. It is noteworthy that gliotoxin effectively suppressed streptococci which are the major pathogens for piscine infection and mortality in aquaculture industry. Gliotoxin also showed strong antibacterial activity against multidrug-resistant human pathogens (MDR) including Enterococcus faecium 5270 and MRSA (methicillin-resistant Staphylococcus aureus) 3089.

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.

Evaluación de la herramienta SaTScan-Whonet para la detección precoz de brotes en infecciones bacterianas en una institución de tercer nivel de atención en Colombia / Evaluation of the SaTScan-Whonet tool for detecting outbreaks of bacterial infection in a tertiary healthcare institution in Colombia

Introducción: Las infecciones asociadas al cuidado de la salud representan un problema de salud pública y la transmisión horizontal supone un incremento en la morbimortalidad y los costos en la atención. La vigilancia activa es costosa y tiene alto riesgo de omitir la detección de brotes, mientras que la virtual (modelos matemáticos) permite la búsqueda sistemática de alertas de brotes. El objetivo de este estudio es evaluar la relación costo-efectividad del uso de la herramienta SaTScan-Whonet para la detección temprana de infecciones bacterianas, comparada con la vigilancia tradicional en una institución de alta complejidad de Colombia. Metodología: En un hospital universitario de alta complejidad se realizó un estudio retrospectivo, se identificó un brote bacteriano, se caracterizó clínicamente y por biología molecular. Se extrajeron las bases de datos de los sistemas automatizados de identificación y susceptibilidad microbiológica. Se realizaron análisis retrospectivos de SaTScan-Whonet, así como simulaciones diarias para el primer semestre de 2011 de manera prospectiva; también se identificó la fecha para la alerta de detección de brote, tanto en la vigilancia activa como en la virtual. Resultados: Se aislaron 4.584 microorganismos en los servicios de hospitalización tanto UCI como no UCI entre 2010 y 2011 (2.288 y 2.296 respectivamente). Por vigilancia activa se notificó un brote por Enterococcus faecium el 28 de marzo de 2011, que fue caracterizado por biología molecular con la presencia del gen Van A, que confiere resistencia a glucopéptidos. Se identificó de manera retrospectiva una alerta de brote para E. faecium entre el 14 de marzo y el 10 de mayo de 2011 con un intervalo de recurrencia de 609.384. En los análisis prospectivos simulados se identificó la primera alerta de brote de esta bacteria el 13 de abril de 2011 con un intervalo de recurrencia de 3.897 (p = 0,0002655). Conclusión: La utilización de dicha herramienta de manera prospectiva no fue superior a la vigilancia activa en cuanto a oportunidad en la detección. Los análisis retrospectivos tuvieron un alto rendimiento diagnóstico y podrían ser de utilidad para los sistemas de vigilancia y control de los entes reguladores.

Background: Healthcare-associated infections represent a public health problem, and horizontal transmission has led to an increase in morbidity and mortality as well as higher health care costs. Active surveillance is expensive and carries high risk of failing to detect outbreaks. Virtual surveillance (mathematical models) allows a systematic search for alerts to outbreaks. The objective of this study is to evaluate the cost-effectiveness of the SaTScan-Whonet tool for the early detection of outbreaks of bacterial infection, compared with traditional surveillance, in an institution of high complexity in Colombia. Methodology: In a university hospital of high complexity a retrospective study was performed, identifying a bacterial outbreak that was characterised clinically and by molecular biology techniques. Databases of automated systems of identification and microbiological susceptibility were extracted. Retrospective analyses were performed using SaTScan-Whonet and daily simulations during the first semester of 2011 in a prospective manner. The date for the alert to the detection of the outbreak for both active and virtual surveillance was also identified. Results: A total of 4,584 microorganisms were isolated both inside and outside the ICU bet-ween 2010 and 2011 (2,288 and 2,296, respectively). An outbreak of Enterococcus faecium was identified by active surveillance on March 28, 2011. Using molecular biology techniques, the outbreak was characterised, showing the presence of the vanA gene, which confers resistance to glycopeptides. An alert to an Enterococcus faecium outbreak was retrospectively identified between March 14 and May 10, 2011 with a recurrence interval of 609,384. The first alert to outbreak for this bacterium was identified in a prospective simulated analysis on April 13, 2011 with a recurrence interval of 3,897 (P=.0002655). Conclusion: The use of such a tool prospectively is not superior to active surveillance in regard to timely detection of bacterial outbreaks. Retrospective analyses have high diagnostic ability and could be very helpful in systems of surveillance and control of regulatory entities.

Correlation between Infective Factors and Antibiotic Resistance in Enterococci Clinical Isolates in West of Iran / 전남의대학술지

The present study was done to scrutinize the possible relation between infective genes and antimicrobial resistance in Enterococcus faecalis and Enterococcus faecium. Considering the fact that the presence of recognized infective determinants among clinical isolates may promote the emergence of infections and persistence of Enterococci in hospital settings, which can lead to an increase in antimicrobial resistance. 175 E. faecalis and 67 E. faecium isolated from clinical specimens were used. The isolates were identified, and then antibiotic susceptibility testing was performed. The MIC of vancomycin and teicoplanin were determined by broth microdilution method. The presence of infective genes esp, hyl and asa₁ was scrutinized using PCR. Of the 280 enterococcal isolates, 175 (62.5%) isolates were identified as E. faecalis, 67 (24%) as E. faecium and 38 (13.5%) as Enterococcus spp. The results of the antibiotic susceptibility testing showed resistance rates of 5% and 73% to vancomycin and teicoplanin in E. faecalis and E. faecium isolates, respectively. The statistical analysis showed that the esp infective gene has significant associations with ciprofloxacin, erythromycin and tetracycline in E. faecium and with chloramphenicol in E. faecalis strains; the hyl with teicoplanin and vancomycin in E. faecium strains; and also asa₁ with vancomycin in E. faecium and with ampicillin and chloramphenicol in E. faecalis strains. Regarding the relationships between virulence genes and antibiotic resistance in strains of E. faecalis and E. faecium, detection of infective factors associated with invasive diseases has become a major issue of concern.

Comparison of a New Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry Platform, ASTA MicroIDSys, With Bruker Biotyper for Species Identification

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, with its accuracy and speed, is widely used for bacterial identification. The ASTA MicroIDSys system (ASTA, Korea) was recently developed for species identification. We compared its performance with that of Bruker Biotyper (Bruker Daltonics, Germany). Microbes were recovered from sputum, urine, and pus samples from patients admitted to a tertiary care hospital in Korea from January to April 2016. Matrix solution (α-cyano-4-hydroxycinnamic acid) was used, and the peptide profiles acquired from the Microflex LT (Bruker Daltonics) and Tinkerbell LT (ASTA) were analyzed by using their respective software. From 5,322 isolates, Bruker Biotyper identified 163 species; fifty species from 4,919 isolates were identified more than 10 times, including Klebsiella pneumoniae (n=571), Acinetobacter baumannii (n=436), Pseudomonas aeruginosa (n=358), Escherichia coli (n=372), Staphylococcus aureus (n=511), S. epidermidis (n=444), Enterococcus faecium (n=262), E. faecalis (n=220), and Candida albicans (n=248). Identical results, confidence scores (≥ 2.0 for Bruker Biotyper), and acceptable scores (≥140 for ASTA MicroIDSys) were obtained for 86.1% of isolates. Of 4,267 isolates, 99.2% showed acceptable scores in both systems. Results from the ASTA MicroIDSys showed good agreement with those from the Bruker Biotyper. The ASTA MicroIDSys could reliably identify clinically important microorganisms.

Distributions of Listeria spp., Bacillus spp., Enterococcus spp., Staphylococcus spp., and Coliforms Isolated from Agricultural Herb Products from the Market

The study was conducted to investigate the distribution of pathogenic bacteria related to agricultural herb products that are sold on the market in South Korea. A survey was conducted on the microbial contamination levels and antibiotic susceptibility of Bacillus cereus (B. cereus) among 194 agricultural herb products on sale in Seoul. Distributions of those isolates were 252 coliforms, 148 Bacillus spp., 75 Enterococcus spp., 10 Staphylococcus spp., and 6 Listeria spp., respectively. The number of B. cereus isolates was 34, Escherichia coli isolates was three, Enterococcus faecium isolate was one, and Enterococcus faecalis isolate was one. Antibiotic susceptibility of B. cereus isolates was tested against 36 kinds of antibiotic susceptibility discs by disc diffusion method. B. cereus isolates were resistant to 20 kinds of antibiotics and semi-resistant to 11 kinds of antibiotics. On the basis of these results, any agricultural herb product can be assumed to be resistant or semi-resistant to the antibiotics used in human. In conclusion, we suggest sanitary control and special management regarding B. cereus contamination in agricultural herb products.