Frecuencia de aislamiento de Staphylococcus spp meticilina resistentes y Enterococcus spp vancomicina resistentes en hospitales de Cuba Frequency of methicilline-resistant Staphylococcus spp and vancomycin-resistant Enterococcus spp isolates in Cuban hospitals
Leonora González Mesa
Full Text Available La resistencia a meticilina en el género Staphylococcus spp es un problema creciente en el ámbito mundial. La producción de una PBP alterada (PBP2a con baja afinidad a betalactámicos, mediada por el gen mec A, es la responsable de esta resistencia. Mientras que los Staphylococcus spp todavía permanecen sensibles a vancomicina, algunos Enterococcus spp han adquirido la capacidad de neutralizar esta droga. En nuestro país no se conocen datos actualizados sobre la tasa de infección por S. aureus meticilina resistente (SAMR, ni sobre la circulación de este germen en la comunidad, tampoco existen reportes de Enterococcus spp vancomicina resistente (EVR. En este estudio fueron analizadas 774 cepas, colectadas en hospitales del país. Se determinó el mecanismo de resistencia utilizando métodos sugeridos por las guías NCCLS. El 9.3 % (23 de los S. aureus aislados en los hospitales y 4.0% (7 S. aureus aislados en la comunidad, fueron SAMR, portadores del gen mec A, el 69.9 % (72 de Staphylococcus coagulasa negativo, fueron resistentes a oxacilina. En la detección del Enterococcus spp vancomicina resistente (EVR, se encontró una cepa portadora de este fenotipo. Nuestros resultados revelan que en nuestro país los SAMR no son un problema en los hospitales, ni en el ambiente comunitario, a pesar de que se reporta por primera vez la circulación de estos en la comunidad y la circulación de EVR en el ambiente hospitalario, su frecuencia es muy baja lo que refleja los avances obtenidos en la aplicación de políticas encaminadas a racionalizar el uso y consumo de antibióticos.Resistance to methicilline in Staphylococcus spp genus is a growing problem worldwide. The production of an altered penicillin-fixing protein with low mecA gen-mediated affinity to beta-lactams is responsible for this resistance. Although Staphylococcus spp still remain susceptible to vancomycin, some Enterococcus spp have acquired the capacity of neutralizing this drug. In
Carlos Andrés Rodríguez
Full Text Available Objetivo. Revisar la evolución y mecanismos moleculares de la resistencia de Staphylococcus aureus a vancomicina. Fuente de los datos. Se consultó la base de datos MEDLINE y se seleccionaron artículos tipo reportes de caso, estudios bioquímicos, de microscopía electrónica y biología molecular pertinentes. Síntesis. Después de casi 40 años de eficacia ininterrumpida de la vancomicina, en 1997 se reportaron los primeros casos de fracaso terapéutico debido a cepas de Staphylococcus aureus con resistencia intermedia, denominadas VISA (concentración inhibitoria mínima, CIM, 8 a 16 ?g/ml, así como a cepas con resistencia heterogénea hVISA (CIM global = 4 ?g/ml, pero con subpoblaciones VISA, en las cuales la resistencia está mediada por engrosamiento de la pared celular y disminución de su entrecruzamiento, lo que afecta la llegada del antibiótico al blanco principal, los monómeros del peptidoglicano en la membrana plasmática. En 2002 se aisló la primera de las 3 cepas reportadas hasta la fecha con resistencia total al antibiótico, denominadas VRSA (CIM>32 ?g/ml, en las que se encontró el transposón Tn1546 proveniente de Enterococcus spp, responsable del reemplazo de la terminación D-Ala-D-Ala por D-Ala-Dlactato en los precursores de la pared celular con pérdida de la afinidad por el glicopéptido. Conclusiones. La resistencia a vancomicina es una realidad en S. aureus, mediada en el caso de VISA por alteraciones en la pared celular que atrapan el antibiótico antes de llegar al sitio de acción, y en el caso de VRSA, por transferencia desde Enterococcus spp. de genes que llevan a la modificación del blanco molecular.
Riboldi, Gustavo Pelicioli; Frazzon, Jeverson; d’Azevedo, Pedro Alves; Frazzon, Ana Paula Guedes
Fifty-six Enterococcus spp. strains were isolated from foods in Southern Brazil, confirmed by PCR and classified as Enterococcus faecalis (27), Enterococcus faecium (23) and Enterococcus spp (6). Antimicrobial susceptibility tests showed resistance phenotypes to a range of antibiotics widely administrated in humans such as gentamycin, streptomycin, ampicillin and vancomycin. PMID:24031330
Endocarditis due to vancomycin-resistant Enterococcus raffinosus successfully treated with linezolid: case report and review of literature Endocarditis por Enterococcus raffinosus resistente a vancomicina exitosamente tratada con linezolid: caso clínico y revisión de la literatura
Full Text Available Enterococcus raffinosus is scarcely found in clinical samples and even less frequently as etiologic agent of endocarditis. We are herein presenting one case of mitral prosthetic-valve endocarditis in a 77-y-o male due to a vancomycinresistant Enterococcus raffinosus isolate, successfully treated with 6 weeks of linezolid, and a two-year follow up.Enterococcus raffinosus es una especie poco frecuente en materiales clínicos y menos aún como agente etiológico de endocarditis. En este trabajo se presenta un caso de endocarditis de válvula mitral protésica en un paciente de 77 años debida a Enterococcus raffinosus resistente a vancomicina y que fue exitosamente tratada con linezolid durante 6 semanas, con un seguimiento de 2 años.
This study investigated the prevalence and antibiotics susceptibility of Enterococcus spp. isolated from patients and some selected hospital environment in Abuja, Nigeria. The samples included clinical and environmental. The clinical samples included stool, urine and wound swabs while the environmental samples ...
Full Text Available During the period from February until March 2004, 108 samples of soft cheese originating from markets of Pancevo, Subotica and Belgrade were examined. Microbiological analyses of the cheese samples to the presence of Escherichia coli was performed using methods described in the Regulations on methods for performing microbiological analyses and super analyses of consumer articles, while the presence of bacteria Enteroccocus spp. was performed on the dexter agar. From 108 samples of soft cheese from the territories of Pancevo, Belgrade and Subotica were isolated: Enterococcus spp. from 96% and Escherichia coli from 69%, cheese samples. Verocytotoxic E.coli was not isolated from any of the taken cheese samples.
Full Text Available Twenty-two bacteriocin-producing Enterococcus isolates obtained from food and animal sources, and demonstrating activity against Listeria monocytogenes, were screened for bacteriocin-related genes using a bacteriocin PCR array based on known enterococcal bacteriocin gene sequences in the NCBI GenBank database. The 22 bacteriocin-positive (Bac+ enterococci included En. durans (1, En. faecalis (4, En. faecium (12, En. hirae (3, and En. thailandicus (2. Enterocin A (entA, enterocins mr10A and mr10B (mr10AB, and bacteriocin T8 (bacA were the most commonly found structural genes in order of decreasing prevalence. Forty-five bacteriocin genes were identified within the 22 Bac+ isolates, each containing at least one of the screened structural genes. Of the 22 Bac+ isolates, 15 possessed two bacteriocin genes, seven isolates contained three different bacteriocins, and three isolates contained as many as four different bacteriocin genes. These results may explain the high degree of bactericidal activity observed with various Bac+ Enterococcus spp. Antimicrobial activity against wild-type L. monocytogenes and a bacteriocin-resistant variant demonstrated bacteriocins having different modes-of-action. Mixtures of bacteriocins, especially those with different modes-of-action and having activity against foodborne pathogens, such as L. monocytogenes, may play a promising role in the preservation of food.
Estudo do perfil de susceptibilidade antimicrobiana e avaliação molecular de amostras de Enterococcus spp isoladas de pacientes hospitalizados - DOI: 10.4025/actascihealthsci.v25i1.2249 Study of drugs susceptibility and molecular samples evaluation of Enterococcus spp isolated in hospitalized patients - DOI: 10.4025/actascihealthsci.v25i1.2249
Celso Luis Cardoso
Full Text Available Neste estudo foi investigado o perfil de susceptibilidade antimicrobiana e avaliação molecular de amostras de Enterococcus spp, isoladas de urina, secreção purulenta, sangue e ferida cirúrgica de pacientes do Hospital Universitário de Maringá, estado do Paraná, no período entre 1998 e 2000. As amostras foram avaliadas quanto à susceptibilidade a drogas, tipificadas através de JB1-PCR e investigadas quanto à presença de gene Van A e Van B. Das 58 amostras, 98,3% foram classificadas como Enterococcus faecalis. Todas as amostras foram sensíveis à nitrofurantoína e vancomicina, 96,6% sensíveis à ampicilina, 32,8% foram resistentes à estreptomicina, 10,3% resistentes à gentamicina e ciprofloxacina e 6,9% resistentes a norfloxacina. Por meio do teste de CIM, para vancomicina, observou-se crescimento bacteriano até a concentração de 4μg/ml. Não foi detectada a presença de gene Van A e Van B. A amplificação por JB1-PCR proporcionou padrões de bandas com até 7 fragmentos sendo as amostras divididas em 6 grupos distintos. Não houve prevalência de algum padrão de bandeamento que denotasse a presença de um clone e não foi possível estabelecer correlação entre genótipo com resistência antimicrobiana.The aim of this study was to investigate the prevalency of Enterococcus spp, isolated from different clinical materials. The strains were evaluated for drugs susceptibility, typing by JB1-PCR and screened to the presence of Van A and Van B gene. From the 58 strains, 98.3% were classified as Enterococcus faecalis. All strains were sensible to nitrofurantoin and vancomycin, 96.6% were sensible to ampicilin, 32.8% were resistant to streptomycin, 10.3% were resistant to gentamicin and ciprofloxacin and 6.9% were resistant to norfloxacin. Using CIM test to check vancomycin susceptibility was observed bacterial growth until 4μg/ml. The presence of Van A e Van B gene was not detected. The amplification by JB1-PCR provided band
Estudio de actinomicetos marinos aislados de la costa central del Perú y su actividad antibacteriana frente a Staphylococcus aureus Meticilina Resistentes y Enterococcus faecalis Vancomicina Resistentes Study of marine actinomycetes isolated from the central coast of Peru and their antibacterial activity against Methicillin-Resistant Staphylococcus aureus and Vancomycin-Resistant Enterococcus faecalis
Full Text Available Objetivos. Determinar el potencial antimicrobiano de actinomicetos marinos frente a cepas S. aureus meticilino-resistentes (MRSA y E. faecalis vancomicina-resistentes (VRE. Materiales y métodos. En dos medios de cultivo se sembraron 29 cepas de actinomicetos aislados de sedimento marino. Se evaluó la capacidad inhibitoria mediante pruebas de antagonismo in vitro para MRSA y VRE. Se procesó los extractos orgánicos de tres actinomicetos seleccionados para determinar la Concentración Mínima Inhibitoria (CMI del compuesto activo. Resultados. La mayoría de los actinomicetos aislados correspondieron a un grupo homogéneo de blanco-grisáceos (62% con buen nivel de crecimiento en agar marino. Los porcentajes inhibitorios fueron superiores a 85% para ambos patógenos con halos de inhibición mayores a 69 y 78 mm de diámetro para MRSA y VRE respectivamente. Los extractos diclorometánicos de tres de los actinomicetos aislados (I-400A, B1-T61, M10-77 mostraron gran potencial inhibitorio de ambos patógenos, siendo M10-77 la cepa de actinomiceto de mayor actividad antibiótica frente a S. aureus ATCC 43300 resistente a meticilina y E. faecalis ATCC 51299 resistente a vancomicina con una Concentración Mínima Inhibitoria (CMI de 7,9 y 31,7 μg/ mL respectivamente. El análisis filogenético de la cepa M10- 77 presenta un 99% de similaridad con la especie marina Streptomyces erythrogriseus. Conclusiones. El sedimento marino de la costa central del Perú es fuente promisorio de cepas de actinomicetos con gran capacidad de producir compuestos bioactivos capaces de inhibir patógenos tipificados como multidrogo-resistentes tales como S. aureus meticilino resistentes y E. faecalis vancomicina resistentes.Objectives. To determine the antimicrobial potential of marine actinomycetes against drug-resistant pathogens represented by strains of methicillin-resistant Staphylococcus aureus (MRSA and vancomycin-resistant Enterococcus faecalis (VRE. Materials and
Lépesová, Kristína; Kraková, Lucia; Pangallo, Domenico; Medveďová, Alžbeta; Olejníková, Petra; Mackuľak, Tomáš; Tichý, Jozef; Grabic, Roman; Birošová, Lucia
Urban wastewater contains different micropollutants and high number of different microorganisms. Some bacteria in wastewater can attach to the surfaces and form biofilm, which gives bacteria advantage in fight against environmental stress. This work is focused on bacterial community analysis in biofilms isolated from influent and effluent sewerage of wastewater treatment plant in Bratislava. Biofilm microbiota detection was performed by culture-independent and culture-dependent approaches. Composition of bacterial strains was detected by denaturing gradient gel electrophoresis fingerprinting coupled with the construction of 16S rRNA clone libraries. The biofilm collected at the inlet point was characterized primarily by the presence of Pseudomonas sp., Acinetobacter sp. and Janthinobacterium sp. clones, while in the biofilm isolated at outflow of wastewater treatment plant members of Pseudomonas genus were largely detected. Beside this analysis prevalence of antibiotics and resistant coliforms, Enterococcus spp. and Staphylococcus spp. in sewerage was studied. In influent wastewater were dominant antibiotics like azithromycin, clarithromycin and ciprofloxacin. Removal efficiency of these antibiotics notably azithromycin and clarithromycin were 30% in most cases. The highest number of resistant bacteria with predominance of coliforms was detected in sample of effluent biofilm. Multidrug resistant strains in effluent biofilm showed very good ability to form biofilm. Copyright © 2018. Published by Elsevier Ltd.
Full Text Available The study aimed at determining the level of resistance of selected bacterial species (Staphylococcus spp., Enterococcus spp., Escherichia coli isolated from rectal swabs of pigs to antimicrobial agents. The tested strains were isolated from piglets aged 7 to 30 days. Bacterial species were identified by standard microbiological techniques and susceptibility to antibiotics was determined quantitatively by the standard microdilution method. Resistance of the Staphylococcus aureus strain to oxacillin was confirmed by detection of the mecA gene and PBP2a. A total of 115 Staphylococcus spp. isolates were collected. In the case of Staphylococcus aureus, the methicillin-resistant strain (MRSA was identified. Moreover, higher frequency of coagulase-negative staphylococci with minimum inhibitory concentration of oxacillin ≥ 0.5 mg/l was noticed. Inducible resistance to clindamycin in the Staphylococcus hominis strain was also detected. The strains of Enterococcus spp. (61 isolates exhibited high resistance to tetracycline (98.5%, erythromycin (86.8% and chloramphenicol (54.4%. Vancomycin-resistant enterococci were not isolated. In the case of Escherichia coli strains (111 isolates, higher frequency of resistant strains to tetracycline (81.1% and ampicillin (62.2% was documented. Resistance to fluoroquinolones and production of broad-spectrum β-lactamases was not noticed. The presented study may be considered as a pilot project assessing the prevalence of resistant bacteria in piglets kept on a single farm. It demonstrated the presence of resistant strains of Staphylococcus spp., including one MRSA strain, Enterococcus spp. and Escherichia coli. These strains may be present as a result of postnatal colonization with both bacterial microflora of dams and environmental microflora.
Rubião, Cynthia Annes; Franco, Robson Maia; Mesquita, Eliana de Fátima Marques de; Miguel, Marco Antonio Lemos; Cabral, Claudius Couto; Fonseca, Ana Beatriz Monteiro
ABSTRACT The aim of this study was to determine and compare the Most Probable Number (MPN) of Total Coliforms (TC), Escherichia coli and Enterococcus spp. and to characterize the antimicrobial resistance profiles of Enterococcus spp. isolated from oysters collected in the Barra de Guaratiba Mangrove, Rio de Janeiro, Brazil. The enumeration of E. coli has been used to indicate fecal contamination and hygienic-sanitary conditions of bivalve molluscs. Enterococci are capable to transfer several ...
Hassan, Reem Mostafa; Ghaith, Doaa Mohammad; Ismail, Dalia Kadry; Zafer, Mai Mahmoud
The incidence of reduced susceptibility to tigecycline (TIG) is increasing. This study aimed to analyse the in vitro activity of TIG against Enterococcus spp. isolates recovered from hospitalised patients and to evaluate the effect of omeprazole on the in vitro antimicrobial activity of TIG against several enterococcal species. A total of 67 Enterococcus clinical isolates were identified by MALDI-TOF/MS and multiplex PCR. Minimum inhibitory concentrations (MICs) of TIG alone and in combination with omeprazole (10, 30 and 60mg/L) were determined by broth microdilution. Antibiotic susceptibility to other antibiotics was determined by disk diffusion. The presence of van, tet(X) and tet(X1) genes was tested by multiplex PCR. Of the 67 Enterococcus isolates, 2 (3.0%) were resistant to TIG and 13 (19.4%) were intermediate-resistant according to EUCAST. The frequencies of resistance to norfloxacin (80.6%), doxycycline (80.6%), levofloxacin (74.6%) and ciprofloxacin (71.6%) were highest, whilst that of vancomycin (25.4%) was lowest. The vanA gene was detected in 11 Enterococcus isolates (8 Enterococcus faecalis, 3 Enterococcus faecium), vanB in 3 Enterococcus isolates (2 E. faecium, 1 E. faecalis) and vanC-2/3 in 3 Enterococcus casseliflavus. Nine isolates (13.4%) were positive for tet(X1). TIG resistance occurred both in patients receiving or not TIG and/or omeprazole. Omeprazole increased TIG MICs by 4-128-fold. The possibility of selection of TIG-non-susceptible Enterococcus in the gut may occur with long-term use of omeprazole. Omeprazole influenced TIG activity in a concentration-dependent manner. To our knowledge; this is the first report of TIG-non-susceptible Enterococcus spp. in Egypt. Copyright © 2017 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.
Bender, Eduardo André; de Freitas, Ana Lúcia Peixoto; Reiter, Keli Cristine; Lutz, Larissa; Barth, Afonso Luís
In the past two decades the members of the genus Enterococcus have emerged as important nosocomial pathogens worldwide. In the present study, we evaluated the antimicrobial resistance and genotypic characteristics of 203 Enterococcus spp. recovered from different clinical sources from two hospitals in Porto Alegre, Rio Grande do Sul, Brazil. The species were identified by conventional biochemical tests and by an automated system. The genetic diversity of E. faecalis presenting high-level aminoglycoside resistance (HLAR) was assessed by pulsed-field gel electrophoresis of chromosomal DNA after SmaI digestion. The E. faecalis was the most frequent specie (93.6%), followed by E. faecium (4.4%). The antimicrobial resistance profile was: 2.5% to ampicillin, 0.5% to vancomycin, 0.5% teicoplanin, 33% to chloramphenicol, 2% to nitrofurantoin, 66.1% to erythromycin, 66.5% to tetracycline, 24.6% to rifampicin, 30% to ciprofloxacin and 87.2% to quinupristin-dalfopristin. A total of 10.3% of the isolates proved to be HLAR to both gentamicin and streptomycin (HLR-ST/GE), with 23.6% resistant only to gentamicin (HLR-GE) and 37.4% only to streptomycin (HLR-ST). One predominant clonal group was found among E. faecalis HLR-GE/ST. The prevalence of resistance among beta-lactam antibiotics and glycopeptides was very low. However, in this study there was an increased number of HLR Enterococcus which may be spreading intra and inter-hospital. PMID:24031416
Cynthia Annes Rubião
Full Text Available ABSTRACT The aim of this study was to determine and compare the Most Probable Number (MPN of Total Coliforms (TC, Escherichia coli and Enterococcus spp. and to characterize the antimicrobial resistance profiles of Enterococcus spp. isolated from oysters collected in the Barra de Guaratiba Mangrove, Rio de Janeiro, Brazil. The enumeration of E. coli has been used to indicate fecal contamination and hygienic-sanitary conditions of bivalve molluscs. Enterococci are capable to transfer several antimicrobial resistance genes to pathogenic bacteria, including those from Gram-negative group. The oysters were bought from local fishermen and a total of 123 individuals were analyzed. The TC, E. coli and Enterococcus spp. MPN mean were 26,300/100 g, 3,260/100 g and 2,820/100 g, respectively. The only correlation found was between TC and E. coli. Two strains of Enterococcus spp. were resistant to three different antimicrobial categories, including a high level resistance to streptomycin. One strain presented intermediate resistance to vancomycin. The E. coli levels exceeded the limits established by international legislation. This microbiological contamination in oysters reflects the water pollution and indicates a probable contamination of other seafood species from this mangrove, which can represent a risk for consumers and a threat to the environment and public health.
Comparative analysis on antibiotic resistance characteristics of Listeria spp. and Enterococcus spp. isolated from laying hens and eggs in conventional and organic keeping systems in Bavaria, Germany.
Schwaiger, K; Schmied, E-M V; Bauer, J
By investigating the prevalence and antimicrobial resistance characteristics of Gram-positive bacteria from organic and conventional keeping systems of laying hens, it was to be determined to what extent these properties are influenced by the different systems. For this purpose, a total of 799 cloacal swabs and 800 egg samples were examined. Prevalences for all selected bacteria from cloacal swabs were much the same for both organic and caged birds: Listeria spp.1.3%[org] versus 1.6%[con]; Enterococcus spp. 95.5%[org] versus 97.5%[con]. Egg contents and eggshells were generally contaminated to a lesser extent, primarily with Enterococcus spp. Listeria isolates were susceptible to almost all tested antibiotics, only three Listeria innocua from conventional keepings were resistant to clindamycin; one isolate additionally to imipenem. High percentages of Enterococcus faecalis were resistant to doxycycline and macrolides. Enterococcus faecium proved to have high resistance rates to clindamycin, fosfomycin and erythromycin; 9.1% were even resistant to the reserve antibiotic synercid. Further, Enterococcus spp. showed higher resistance rates to doxycycline, erythromycin, fosfomycin and rifampicin. No glycopeptide resistant enterococci were detected. A correlation between keeping system and resistance/susceptibility rates could be demonstrated. In detail, E. faecalis from organic laying hen husbandries showed significant lower resistance prevalences to tylosin, streptomycin and doxycycline; susceptibility rates were higher for enrofloxacin and ciprofloxacin. Rifampicin and imipenem were more effective in isolates from conventional keepings (P < 0.05). The amounts of resistant isolates of the Enterococcus raffinosus from organic farms were significantly lower, the amounts of sensitive isolates were significantly higher than from conventional farms concerning eight antibiotics (P < 0.05). When comparing the susceptibility/resistance rates, as well as the mean minimum
Estudio de la susceptibilidad a vancomicina de cepas de Staphyloccus spp., aisladas del hospital Antonio Patricio de Alcalá, Cumaná, estado Sucre | Vancomycin susceptibility study of Staphylococcus spp. Strains isolated from Antonio Patricio de Alcalá hospital, Cumaná, Sucre state
Full Text Available De todas las cepas de Staphylocccus aureus almacenadas en el cepario del Laboratorio de Resistencia Bacteriana del IIBCAUDO, se analizaron 14 para determinar la disminución de susceptibilidad a vancomicina. Sesenta y cuatro por ciento de las cepas crecieron en el cribado de vancomicina. La CMI a vancomicina de las cepas estuvo entre 1 y 2 mg/L por microdilución. La CMI por dilución en agar fue entre 2 y 4 mg/L. La CMB de las cepas fue de 2 y 4 mg/L. Ninguna cepa fue tolerante. No se detectaron cepas hVISA por PAP-AUC. Se recomienda realizar el cribado de vancomicina en cepas aisladas de sitios estériles.
Colonización por cepas de Enterococcus spp. VanA en pacientes del hospital universitario Manuel Núñez Tovar, Maturín, Venezuela | Colonization by strains of Enterococcus spp. VanA in patients of the university hospital Manuel Núñez Tovar, Maturín, Venezuela
Full Text Available En el Hospital Universitario Manuel Núñez Tovar de Maturín (HUMNT, estado Monagas, se aislaron tres cepas de Enterococcus faecium VanA en la unidad de cuidados intensivos (UCI y una cepa de E. faecalis VanA en la unidad de hemodiálisis (HD. La prevalencia de colonización de Enterococcus vancomicino resistentes (EVR fenotipo VanA durante el período de estudio fue de 3% en los pacientes de HD y 9% en los de UCI. Los factores de riesgo para los pacientes fueron el uso indiscriminado de antibióticos como vancomicina y cefalosporinas de tercera generación y el tiempo de permanencia en UCI. Existe alto riesgo de infecciones endógenas por cepas EVR en el HUMNT.
Pesavento, G; Calonico, C; Ducci, B; Magnanini, A; Lo Nostro, A
Food specimens were analyzed in order to research Enterococcus spp.: 636 samples of raw meat (227 beef, 238 poultry, and 171 pork), 278 samples of cheese (110 fresh soft cheese and 168 mozzarella cheese), 214 samples of ready-to-eat salads, and 187 samples of ham. 312 strains of Enterococcus spp samples were isolated, then identified and submitted to susceptibility tests against 11 antimicrobial agents. The predominant species were Enterococcus faecalis in raw meat and Enterococcus faecium in retail products. Low percentages of microorganisms were resistant to vancomycin (3.53%), teicoplanin (2.24%), linezolid (0.32%), and amoxicillin in combination with clavulanic acid (0.32%). A high percentage of resistance was noted in E. faecalis at high level gentamicin (21.9%) and tetracycline (60.6%). In general, strains of E. faecalis were more resistant than E. faecium. Enterococci should be considered not only potential pathogens, but also a reservoir of genes encoding antibiotic resistance which can be transferred to other microorganisms. Continuous monitoring of their incidence and emerging resistance is important in order to identify foods which potentially represent a real risk to the population, and to ensure effective treatment of human enterococcal infections. Copyright © 2014 Elsevier Ltd. All rights reserved.
Full Text Available Introducción. Actualmente se considera a Enterococcus spp. como uno de los agentes de infección hospitalaria más importantes, siendo su resistencia a los antibióticos un problema importante en los centros de salud. Objetivos. Caracterizar la resistencia antimicrobiana en 50 cepas de Enterococcus spp. aisladas de muestras clínicas de pacientes hospitalizados. Materiales y métodos. Se llevó a cabo un estudio de tipo descriptivo observacional de corte transversal en 50 aislamientos clínicos de estas especies microbianas. Se trabajó un aislamiento por paciente. La identificación y la sensibilidad a los antibióticos se realizaron por métodos automatizados y convencionales. El análisis fenotípico de los mecanismos de resistencia a glucopéptidos se hizo según las recomendaciones de la Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. Resultados. De 50 aislamientos, 30 (60,0 % y 20 (40,0 % pertenecían a las especies de Enterococcus faecalis y Enterococcus faecium, respectivamente. La resistencia global expresada por este género fue de 38/50 (76,0 % para ampicilina; 33/50 (66,0 % para gentamicina de alto nivel; 34/50 (68,0 % para estreptomicina de alto nivel; 26/50 (52,0 % para ciprofloxacina; 4/50 (8,0 % para linezolid; 17/50 (34,0 % para teicoplanina; 25/50 (50,0 % para vancomicina; 31/50 (62,0 % para minociclina; 34/50 (68,0 % para tetraciclina y 9/50 (18,0 % para nitrofurantoina. Frente a los glucopéptidos, 25/50 (50,0 % y 10/50 (20,0 % de los aislamientos presentaron los mecanismos Van A y Van B, respectivamente. Conclusiones. Podemos concluir que la mayoría de las veces, las cepas aisladas en el Hospital Hermanos Ameijeiras mostraron porcentajes de resistencia por encima de lo reportado en la literatura científica consultada. El alto porcentaje de cepas con resistencia a la vancomicina podría influir en la aparición de otros gérmenes Gram positivos con resistencia a este fármaco. Se reporta por
Marrow, Judilee; Whittington, Julia K; Mitchell, Mark; Hoyer, Lois L; Maddox, Carol
Due to their predatory nature, raptor species may serve as important indicators of environmental contamination with antimicrobial-resistant bacteria. Raptors prey on small rodents and birds that have diverse habitat ranges, including urban and rural environments, and their intestinal microflora can reflect that of the animals on which they feed. Enterococcus spp. were selected as target organisms because they have been isolated from the avian gastrointestinal tract, can be conferred by prey items, and because they are capable of multiple resistance patterns. They are also a concerning source of human antimicrobial resistance. In this study fecal cultures were obtained from 15 May 2004 to 31 August 2004, from 21 free-living raptors and four captive raptors. Enterococcus was isolated from 21 (84%) of the 25 birds, and 54 isolates were chosen for further study based upon unique colony morphology. The most common isolate recovered was Enterococcus faecalis (95%, 95% confidence interval [CI]: 89-100). One bird in the study was determined to have Enterococcus gallinarum. Two distinct ribotypes of E. faecalis were identified, one with unique bands at 11 and 13 kb and the other with unique bands at 14 and 20 kb. Both ribotypes were found in free-living and captive birds. The Enterococcus isolates in this study demonstrated a variety of antimicrobial-resistance characteristics, including almost complete resistance to amikacin, first-generation cephalosporins, spectinomycin, and sulphadimethoxime. Isolates demonstrated variable resistance to chloramphenicol, gentamicin, enrofloxacin, erythromycin, and ticarcillin. No phenotypically vancomycin-resistant E. faecalis isolates were recovered from any of the raptors; three isolates had intermediate level susceptibility. A significantly higher number of isolates collected from captive birds demonstrated resistance to chloramphenicol than those obtained from free-living birds. This trend was not duplicated with any of the remaining
Hombach, Michael; Jetter, Marion; Blöchliger, Nicolas; Kolesnik-Goldmann, Natalia; Keller, Peter M; Böttger, Erik C
Abstract Background We investigated the feasibility of rapid disc diffusion antibiotic susceptibility testing (rAST) with reading of inhibition zones after 6 and/or 8 h of incubation for Enterococcus faecalis, Enterococcus faecium, Pseudomonas aeruginosa and Acinetobacter baumannii. In addition, we evaluated discrimination of resistant populations from the WT populations at early timepoints and the requirement for clinical breakpoint adaptations for proper interpretation of rAST data. Methods In total, 815 clinical strains [E. faecalis (n = 135), E. faecium (n = 227), P. aeruginosa (n = 295) and A. baumannii (n = 158)] were included in this study. Disc diffusion plates were streaked, incubated and imaged using the WASPLabTM automation system. WT populations and non-WT populations were defined using epidemiological cut-offs. Results and conclusions rAST at 6 and 8 h was possible for A. baumannii and enterococci with readability of inhibition zones >90%. Overall categorical agreement of rAST at 6 h with AST at 18 h was 97.2%, 97.4% and 95.3% for E. faecalis, E. faecium and A. baumannii, respectively. With few exceptions, major categorization error rates were <1% for A. baumannii, and vancomycin-resistant E. faecium were clearly separated from the WT at 6 h. For P. aeruginosa the average readability of inhibition zones was 68.9% at 8 h and we found an overall categorical agreement of 94.8%. Adaptations of clinical breakpoints and/or introduction of technical buffer zones, preferably based on aggregated population data from various epidemiological settings, are required for proper interpretation of rAST. PMID:29186434
Boulianne, Martine; Arsenault, Julie; Daignault, Danielle; Archambault, Marie; Letellier, Ann; Dutil, Lucie
An observational study was conducted of chicken and turkey flocks slaughtered at federal processing plants in the province of Quebec, Canada. The objectives were to estimate prevalence of drug use at hatchery and on farm and to identify antimicrobial resistance (AMR) in cecal Escherichia coli and Enterococcus spp. isolates and factors associated with AMR. Eighty-two chicken flocks and 59 turkey flocks were sampled. At the hatchery, the most used antimicrobial was ceftiofur in chickens (76% of...
Martín, B; Corominas, L; Garriga, M; Aymerich, T
Four local small-scale factories were studied to determine the sources of enterococci in traditional fermented sausages. Different points during the production of a traditional fermented sausage type (fuet) were evaluated. Randomly amplified polymorphic DNA (RAPD)-PCR was used to type 596 Enterococcus isolates from the final products, the initial meat batter, the casing, the workers' hands and the equipment. Species-specific PCR-multiplex and the partial sequencing of atpA gene and 16S rRNA gene sequencing allowed the identification of the isolates: Enterococcus faecalis (31.4%), Enterococcus faecium (30.7%), Enterococcus sanguinicola (14.9%), Enterococcus devriesei (9.7%), Enterococcus malodoratus (7.2%), Enterococcus gilvus (1.0%), Enterococcus gallinarum (1.3%), Enterococcus casseliflavus (3.4%), Enterococcus hermanniensis (0.2%), and Enterococcus durans (0.2%). A total of 92 different RAPD-PCR profiles were distributed among the different factories and samples evaluated. Most of the genotypes found in fuet samples were traced back to their source. The major sources of enterococci in the traditional fermented sausages studied were mainly the equipment followed by the raw ingredients, although a low proportion was traced back to human origin. This work contributes to determine the source of enterococcal contamination in fermented sausages and also to the knowledge of the meat environment.
Carey, Stephanie Ann; Goldstein, Rachel E. Rosenberg [Maryland Institute for Applied Environmental Health, University of Maryland School of Public Health, College Park, MD (United States); Gibbs, Shawn G. [Department of Environmental Health, School of Public Health-Bloomington, Indiana University, Bloomington, IN (United States); Claye, Emma [Maryland Institute for Applied Environmental Health, University of Maryland School of Public Health, College Park, MD (United States); He, Xin [Department of Epidemiology and Biostatistics, University of Maryland School of Public Health, College Park, MD (United States); Sapkota, Amy R., E-mail: email@example.com [Maryland Institute for Applied Environmental Health, University of Maryland School of Public Health, College Park, MD (United States)
Reclaiming municipal wastewater for agricultural, environmental, and industrial purposes is increasing in the United States to combat dwindling freshwater supplies. However, there is a lack of data regarding the microbial quality of reclaimed water. In particular, no previous studies have evaluated the occurrence of vancomycin-resistant enterococci (VRE) in reclaimed water used at spray irrigation sites in the United States. To address this knowledge gap, we investigated the occurrence, concentration, and antimicrobial resistance patterns of VRE and vancomycin-susceptible enterococci at three U.S. spray irrigation sites that use reclaimed water. We collected 48 reclaimed water samples from one Mid-Atlantic and two Midwest spray irrigation sites, as well as their respective wastewater treatment plants, in 2009 and 2010. Samples were analyzed for total enterococci and VRE using standard membrane filtration. Isolates were purified and then confirmed using biochemical tests and PCR. Antimicrobial susceptibility testing was conducted using the Sensititre® microbroth dilution system. Data were analyzed by two-sample proportion tests and one-way analysis of variance. We detected total enterococci and VRE in 71% (34/48) and 4% (2/48) of reclaimed water samples, respectively. Enterococcus faecalis was the most common species identified. At the Mid-Atlantic spray irrigation site, UV radiation decreased total enterococci to undetectable levels; however, subsequent storage in an open-air pond at this site resulted in increased concentrations of enterococci. E. faecalis isolates recovered from the Mid-Atlantic spray irrigation site expressed intrinsic resistance to quinupristin/dalfopristin; however, non-E. faecalis isolates expressed resistance to quinupristin/dalfopristin (52% of isolates), vancomycin (4%), tetracycline (13%), penicillin (4%) and ciprofloxacin (17%). Our findings show that VRE are present in low numbers in reclaimed water at point-of-use at the sampled spray
Poulsen, Louise Ladefoged; Bisgaard, Magne; Son, Nguyen Thai; Trung, Nguyen Vu; An, Hoang Manh; Dalsgaard, Anders
Urinary tract infections (UTI) are one of the most common infections among women worldwide. E. coli often causes more than 75% of acute uncomplicated UTI, however, little is known about how recurrent UTIs and indiscriminate use of antimicrobials affect the aetiology of UTIs. This study aimed to establish the aetiology of UTI in a population of recurrent and self-medicated patients referred from pharmacies to a hospital in Hanoi, Vietnam and to describe genotypes and antimicrobial susceptibility of the associated bacterial pathogens. The aetiology of bacterial pathogens associated with UTI (defined as ≥ 104 CFU/ml urine) was established by phenotypic and molecular methods. Enterococcus faecalis isolates were typed by Multi Locus Sequence Typing (MLST), Pulsed-Field Gel Electrophoresis (PFGE) and antimicrobial susceptibility testing. Urine samples from 276 patients suffering symptoms of urinary tract infection were collected and cultured on Flexicult agar® allowing for detection of the most common urine pathogens. Patients were interviewed about underlying diseases, duration of symptoms, earlier episodes of UTI, number of episodes diagnosed by doctors and treatment in relation to UTI. All tentative E. faecalis and E. faecium isolates were identified to species level by PCR, 16S rRNA and partial sequencing of the groEL gene. E. faecalis isolates were further characterized by Multi Locus Sequence Typing and antimicrobial susceptibility testing. Mean age of 49 patients was 48 yrs (range was 11-86 yrs) and included 94% women. On average, patients reported to have suffered from UTI for 348 days (range 3 days-10 years, and experienced 2.7 UTIs during the previous year). Cephalosporins were reported the second drug of choice in treatment of UTI at the hospital. E. faecalis (55.1%), E. coli (12.2%) and Streptococcus gallolyticus subsp. pasteurianus (8.2%) were main bacterial pathogens. MIC testing of E. faecalis showed susceptibility to ampicillin, penicillin and
Carey, Stephanie Ann; Goldstein, Rachel E. Rosenberg; Gibbs, Shawn G.; Claye, Emma; He, Xin; Sapkota, Amy R.
Reclaiming municipal wastewater for agricultural, environmental, and industrial purposes is increasing in the United States to combat dwindling freshwater supplies. However, there is a lack of data regarding the microbial quality of reclaimed water. In particular, no previous studies have evaluated the occurrence of vancomycin-resistant enterococci (VRE) in reclaimed water used at spray irrigation sites in the United States. To address this knowledge gap, we investigated the occurrence, concentration, and antimicrobial resistance patterns of VRE and vancomycin-susceptible enterococci at three U.S. spray irrigation sites that use reclaimed water. We collected 48 reclaimed water samples from one Mid-Atlantic and two Midwest spray irrigation sites, as well as their respective wastewater treatment plants, in 2009 and 2010. Samples were analyzed for total enterococci and VRE using standard membrane filtration. Isolates were purified and then confirmed using biochemical tests and PCR. Antimicrobial susceptibility testing was conducted using the Sensititre® microbroth dilution system. Data were analyzed by two-sample proportion tests and one-way analysis of variance. We detected total enterococci and VRE in 71% (34/48) and 4% (2/48) of reclaimed water samples, respectively. Enterococcus faecalis was the most common species identified. At the Mid-Atlantic spray irrigation site, UV radiation decreased total enterococci to undetectable levels; however, subsequent storage in an open-air pond at this site resulted in increased concentrations of enterococci. E. faecalis isolates recovered from the Mid-Atlantic spray irrigation site expressed intrinsic resistance to quinupristin/dalfopristin; however, non-E. faecalis isolates expressed resistance to quinupristin/dalfopristin (52% of isolates), vancomycin (4%), tetracycline (13%), penicillin (4%) and ciprofloxacin (17%). Our findings show that VRE are present in low numbers in reclaimed water at point-of-use at the sampled spray
a diciembre de 1999 hecho en el Instituto Nacional de Pediatría en la Ciudad de México, se procesaron 97 cepas de Enterococcus (60 de Enterococcus faecalis y 37 de Enterococcus faecium, contra 11 antibióticos. La prueba de susceptibilidad se elaboró con agar, de acuerdo con los estándares del Comité Nacional para el Laboratorio Clínico (NCCLS. Todos los aislamientos fueron probados para determinar la resistencia elevada en contra de beta-lactámicos, aminoglucósidos y glicopéptidos. Asimismo, se determinó el fenotipo de resistencia hacia la vancomicina. Se evaluaron diferencias de proporciones con ji2 o prueba exacta de Fisher. RESULTADOS: La resistencia en general hacia los antibióticos probados fue 17/97 (17.5% a penicilina, ampicilina, amoxicilina-clavulanato e imipenem. No se encontró resistencia elevada ni presencia de producción de beta-lactamasas; 74/97 (48.4% fueron resistentes a eritromicina, 60% resistentes a ciprofloxacina, 31/97 (32% resistentes a gentamicina y 55/97 (56.7% resistentes a estreptomicina. Siete cepas fueron resistentes a vancomicina, todas ellas E. faecium; 5/7 con el fenotipo A y 2/7 con el fenotipo B. Todas las cepas aisladas fueron susceptibles al linezolid. La diferencia en la susceptibilidad antimicrobiana entre las especies fue significativa. CONCLUSIONES: La resistencia antimicrobiana múltiple de Enterococcus spp. es un problema real y es necesaria su vigilancia. El laboratorio de microbiología es la primera línea de defensa en contra de la diseminación de enterococos con resistencia múltiple en el ambiente hospitalario. Todas las cepas aisladas deberían ser probadas en contra de ampicilina, estreptomicina, gentamicina y glicopéptidos.
Boulianne, Martine; Arsenault, Julie; Daignault, Danielle; Archambault, Marie; Letellier, Ann; Dutil, Lucie
An observational study was conducted of chicken and turkey flocks slaughtered at federal processing plants in the province of Quebec, Canada. The objectives were to estimate prevalence of drug use at hatchery and on farm and to identify antimicrobial resistance (AMR) in cecal Escherichia coli and Enterococcus spp. isolates and factors associated with AMR. Eighty-two chicken flocks and 59 turkey flocks were sampled. At the hatchery, the most used antimicrobial was ceftiofur in chickens (76% of flocks) and spectinomycin in turkeys (42% of flocks). Virginiamycin was the antimicrobial most frequently added to the feed in both chicken and turkey flocks. At least 1 E. coli isolate resistant to third-generation cephalosporins was present in all chicken flocks and in a third of turkey flocks. Resistance to tetracycline, streptomycin, and sulfisoxazole was detected in > 90% of flocks for E. coli isolates. Antimicrobial resistance (AMR) was observed to bacitracin, erythromycin, lincomycin, quinupristin-dalfopristin, and tetracycline in both chicken and turkey flocks for Enterococcus spp. isolates. No resistance to vancomycin was observed. The use of ceftiofur at hatchery was significantly associated with the proportion of ceftiofur-resistant E. coli isolates in chicken flocks. In turkey flocks, ceftiofur resistance was more frequent when turkeys were placed on litter previously used by chickens. Associations between drug use and resistance were observed with tetracycline (turkey) in E. coli isolates and with bacitracin (chicken and turkey), gentamicin (turkey), and tylosin (chicken) in Enterococcus spp. isolates. Further studies are needed to provide producers and veterinarians with alternative management practices and tools in order to reduce the use of antimicrobial feed additives in poultry.
Macedo, Ana Sofia
Trabalho apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas. A água não tratada é constantemente desvalorizada como fonte de resistências a antibióticos em países desenvolvidos. Para averiguar este problema de saúde pública, neste trabalho isolaram-se Enterococcus spp. como bactérias indicadoras de contaminação fecal de poços, fontes e minas que fornecem água a diversas comunidades em Portugal e caracterizou-se o s...
Roberts, Marilyn C; No, David B; Marzluff, John M; Delap, Jack H; Turner, Robert
Vancomycin-resistant enterococci [VRE] have been isolated from municipal, hospital and agricultural wastewater, recreational beaches, wild animals, birds and food animals around the world. In this study, American crows (Corvus brachyrhynchos) from sewage treatment plants (WWTP), dairy farms, and a large roost in a restored wetland with corresponding environmental samples were cultured for VRE. A total of 245 samples [156 crows, 89 environmental] were collected and screened for acquired vanA, vanB and/or intrinsic vanC1 genes. Samples were enriched overnight in BHI supplemented with 20μg/mL aztreonam, 4μg/mL vancomycin and plated on m-Enterococcus agar media supplemented with 6μg/mL vancomycin. Selected colonies were grown on BHI media supplemented with 18μg/mL vancomycin. Of these, 24.5% of the crow and 55% the environmental/cow samples were VRE positive as defined by Enterococcus spp. able to grow on media supplemented with 18μg/mL vancomycin. A total of 122 VRE isolates, 43 crow and 79 environmental isolates were screened, identified to species level using 16S sequencing and further characterized. Four vanA E. faecium and multiple vanC1 E. gallinarum were identified from crows isolated from three sites. E. faecium vanA and E. gallinarum vanC1 along with other Enterococcus spp. carrying vanA, vanB, vanC1 were isolated from three environments. All enterococci were multidrug resistant. Crows were more likely to carry vanA E. faecium than either the cow feces or wetland waters/soils. Comparing E. gallinarum vanC1 from crows and their environment would be useful in determining whether crows share VRE strains with their environment. Copyright © 2016 Elsevier B.V. All rights reserved.
Caballero-Granado, F J; Cisneros, J M; Luque, R; Torres-Tortosa, M; Gamboa, F; Díez, F; Villanueva, J L; Pérez-Cano, R; Pasquau, J; Merino, D; Menchero, A; Mora, D; López-Ruz, M A; Vergara, A
A prospective, multicenter study was carried out over a period of 10 months. All patients with clinically significant bacteremia caused by Enterococcus spp. were included. The epidemiological, microbiological, clinical, and prognostic features and the relationship of these features to the presence of high-level resistance to gentamicin (HLRG) were studied. Ninety-three patients with enterococcal bacteremia were included, and 31 of these cases were caused by HLRG (33%). The multivariate analysis selected chronic renal failure, intensive care unit stay, previous use of antimicrobial agents, and Enterococcus faecalis species as the independent risk factors that influenced the development of HLRG. The strains with HLRG showed lower levels of susceptibility to penicillin and ciprofloxacin. Clinical features (except for chronic renal failure) were similar in both groups of patients. HLRG did not influence the prognosis for patients with enterococcal bacteremia in terms of either the crude mortality rate (29% for patients with bacteremia caused by enterococci with HLRG and 28% for patients not infected with strains with HLRG) or the hospital stay after the acquisition of enterococcal bacteremia. Hemodynamic compromise, inappropriate antimicrobial therapy, and mechanical ventilation were revealed in the multivariate analysis to be the independent risk factors for mortality. Prolonged hospitalization was associated with the nosocomial acquisition of bacteremia and polymicrobial infections.
Maicon Marvila Miranda
Full Text Available A Unidade de Terapia Intensiva, local onde são internados pacientes com maior gravidade, é uma das unidades do hospital com maior índice de infecções hospitalares. Os Enterococcus são capazes também de exibir diferentes mecanismos de resistência adquirida a uma ampla variedade de antimicrobianos. Esta pesquisa teve como objetivo identificar a prevalência de cepas de Enterococcus spp. na UTI de um hospital em Cachoeiro de Itapemirim-ES, no período de 2014 à 2015, e determinar sua multirresistência aos antimicrobianos. Os dados foram coletados através do acervo documental do laboratório de análises clínicas. A maior prevalência da bactéria foi no gênero masculino e em amostras de urina foram fonte majoritária de isolamento. Fora verificado alta taxa de resistência a quinolonas, aminoglicosídeos e glicopeptídeos. A necessidade de uma maior vigilância no controle de infecção hospitalar com o intuito de minimizar e/ou eliminar as infecções de pacientes internados, principalmente em UTI.
Full Text Available A case control study was performed in the Parco Nazionale dei Monti Sibillini, Italy, to find out whether roe deer (Capreolus capreolus and red deer (Cervus elaphus were more likely to harbour antibiotic resistant Escherichia coli in their faeces, compared to Enterococcus spp. Ten areas were selected and samples were collected during a fourmonths (May to August, 2008 sampling period. Samples of water (n=12 and feces (n=59, collected at 10 different sites, were cultured for E. coli and Enterococcus spp. The resulting colonies were screened for tetracycline, ampicillin and kanamycin resistance using the Lederberg Replica Plating method (breakpoint 4 μg/ml. All resistant isolates were then selected, and subjected to the CLSI antimicrobial plate susceptibility test (7. Among the water specimens contaminated by E. coli, 80% were found to be resistant to ampicillin, 80% to tetracycline and 40% to kanamycin. Among the water specimens contaminated by Enterococcus spp., 14.29% were found to be resistant to ampicillin, 14.29% to tetracycline and 71.3% to kanamycin. Among the 39 strains of E. coli isolated from red deer feces, 12 were resistant to ampicillin (30.77%, 5 to tetracycline (12,82% and 3 to kanamycin (7.69%. Among the 19 strains of Enterococcus spp. isolated from red deer feces, 0 were resistant to ampicillin (0%, 1 to tetracycline (5.26% and 19 to kanamycin (100. These are significant findings, indicating that antibiotic resistance can be found in naïve animal populations and that red deer and fallow deer could act as sentinels for antimicrobial resistance. Key words Antibiotic-resistance, red deer, fallow deer, Escherichia
Woods, Stephanie E; Lieberman, Mia T; Lebreton, Francois; Trowel, Elise; de la Fuente-Núñez, César; Dzink-Fox, Joanne; Gilmore, Michael S; Fox, James G
Nonhuman primates are commonly used for cognitive neuroscience research and often surgically implanted with cephalic recording chambers for electrophysiological recording. Aerobic bacterial cultures from 25 macaques identified 72 bacterial isolates, including 15 Enterococcus faecalis isolates. The E. faecalis isolates displayed multi-drug resistant phenotypes, with resistance to ciprofloxacin, enrofloxacin, trimethoprim-sulfamethoxazole, tetracycline, chloramphenicol, bacitracin, and erythromycin, as well as high-level aminoglycoside resistance. Multi-locus sequence typing showed that most belonged to two E. faecalis sequence types (ST): ST 4 and ST 55. The genomes of three representative isolates were sequenced to identify genes encoding antimicrobial resistances and other traits. Antimicrobial resistance genes identified included aac(6')-aph(2"), aph(3')-III, str, ant(6)-Ia, tetM, tetS, tetL, ermB, bcrABR, cat, and dfrG, and polymorphisms in parC (S80I) and gyrA (S83I) were observed. These isolates also harbored virulence factors including the cytolysin toxin genes in ST 4 isolates, as well as multiple biofilm-associated genes (esp, agg, ace, SrtA, gelE, ebpABC), hyaluronidases (hylA, hylB), and other survival genes (ElrA, tpx). Crystal violet biofilm assays confirmed that ST 4 isolates produced more biofilm than ST 55 isolates. The abundance of antimicrobial resistance and virulence factor genes in the ST 4 isolates likely relates to the loss of CRISPR-cas. This macaque colony represents a unique model for studying E. faecalis infection associated with indwelling devices, and provides an opportunity to understand the basis of persistence of this pathogen in a healthcare setting.
Stephanie E Woods
Full Text Available Nonhuman primates are commonly used for cognitive neuroscience research and often surgically implanted with cephalic recording chambers for electrophysiological recording. Aerobic bacterial cultures from 25 macaques identified 72 bacterial isolates, including 15 Enterococcus faecalis isolates. The E. faecalis isolates displayed multi-drug resistant phenotypes, with resistance to ciprofloxacin, enrofloxacin, trimethoprim-sulfamethoxazole, tetracycline, chloramphenicol, bacitracin, and erythromycin, as well as high-level aminoglycoside resistance. Multi-locus sequence typing showed that most belonged to two E. faecalis sequence types (ST: ST 4 and ST 55. The genomes of three representative isolates were sequenced to identify genes encoding antimicrobial resistances and other traits. Antimicrobial resistance genes identified included aac(6'-aph(2", aph(3'-III, str, ant(6-Ia, tetM, tetS, tetL, ermB, bcrABR, cat, and dfrG, and polymorphisms in parC (S80I and gyrA (S83I were observed. These isolates also harbored virulence factors including the cytolysin toxin genes in ST 4 isolates, as well as multiple biofilm-associated genes (esp, agg, ace, SrtA, gelE, ebpABC, hyaluronidases (hylA, hylB, and other survival genes (ElrA, tpx. Crystal violet biofilm assays confirmed that ST 4 isolates produced more biofilm than ST 55 isolates. The abundance of antimicrobial resistance and virulence factor genes in the ST 4 isolates likely relates to the loss of CRISPR-cas. This macaque colony represents a unique model for studying E. faecalis infection associated with indwelling devices, and provides an opportunity to understand the basis of persistence of this pathogen in a healthcare setting.
Juan Aguilar, Marta
Vancomicina es un antibiótico glucopéptido, ampliamente utilizado en el tratamiento de la sepsis neonatal nosocomial, que ha mostrado una amplia variabilidad interindividual en los parámetros farmacocinéticos, particularmente en neonatos. Esta situación ha favorecido que vancomicina sea uno de los fármacos más estudiados utilizando diferentes aproximaciones de farmacocinética poblacional con el objetivo de caracterizar sus parámetros farmacocinéticos, identificar los factores individuales que...
Cardoso, María de las Mercedes Elisa; Cardoso, María de las Mercedes Elisa
Las bacterias ácido lácticas, que gozan del status de “Generalmente Reconocidas Como Seguras”, otorgado por la Organización Mundial de la Salud, pueden producir sustancias antimicrobianas de naturaleza proteica, denominadas bacteriocinas, que actúan como bioconservantes alimentarios. Se investigó la capacidad bacteriocinogénica de la cepa de Enterococcus faecalis DBFIQ E24, aislada de leche cruda producida en la región santafesina. Esta cepa fue sensible a vancomicina y carente de capacidad...
Santos Tedim Sousa Pedrosa, Ana Sofía
La prevalencia de infecciones hospitalarias causadas por especies del género Enterococcus ha sido baja desde su descripción como patógenos oportunistas al inicio del siglo XX, hasta finales de los años 70s, coincidiendo con la aparición de las primeras cepas resistentes a antibióticos. La especie Enterococcus faecium es actualmente uno de los principales patógenos nosocomiales debido en parte a la alta prevalencia de cepas resistentes a ampicilina y vancomicina. La estructura poblacional de E...
Análisis clínico-epidemiológico de la portación intestinal de enterococos resistentes a vancomicina en una unidad de terapia intensiva Clinical and epidemiologic analysis of intestinal tract colonization with vancomycin-resistant enterococci in an intensive care unit
Full Text Available En un período de cinco meses y 25 días se investigó la portación intestinal de enterococos resistentes a vancomicina (EVR. Se estudiaron 124 pacientes (73% de 171 admitidos en la unidad de terapia intensiva (UTI, 35 de los cuales (28% resultaron ser portadores. Los aislamientos de EVR (n=35 fueron identificados como Enterococcus faecium (n=18, Enterococcus gallinarum (n=16 y Enterococcus raffinosus (n=1. Todos los aislamientos estudiados fueron resistentes a vancomicina (VAN (CIM90= 512 µg/ml y teicoplanina (CIM90= 32 µg/ml y portaban el gen vanA. Los estudios de tipificación molecular mostraron un alto grado de homología entre los aislamientos de E. faecium (un clon dominante y E. gallinarum (dos tipos clonales, sugiriendo su diseminación a modo de brote. No se encontraron diferencias significativas con la edad y el sexo de los pacientes no portadores (p>0,05, pero si con el tiempo de hospitalización y el uso de esquemas antibióticos de amplio espectro (pIntestinal tract colonization with vancomycin resistant enterococci (VRE was studied during five months and 25 days. Out of 171 patients hospitalized in the intensive care unit, 124 (73% were included in this study. Thirty five of them (28% were recognized as colonized with VRE. VRE isolates (n = 35 were identified as Enterococcus faecium (n=18, Enterococcus gallinarum (n=16, and Enterococcus raffinosus (n=1. All of them were resistant to vancomycin (MIC90= 512 µg/ml and to teicoplanin (MIC90= 32 µg/ml, having the vanA gene. By means of molecular methods a high homology was found among E. faecium and E. gallinarum isolates, respectively, suggesting their spread as a kind of outbreak. No significant differences in age or sex were found among colonized and non-colonized patients (p>0.05. On the other hand, the hospitalization time and the use of broad-spectrum antibiotics were associated with colonization. From this study we highlight the importance of enhancing all measures of
Ana Claudia F. Borges de Campos
Full Text Available O objetivo deste trabalho foi realizar o isolamento e analisar o perfil de resistência antimicrobiana de Enterococcus de carcaças de frango resfriadas e congeladas comercializadas no Distrito Federal, detectando genes de resistência antimicrobiana e identificando as espécies Enterococcus faecalis e Enterococcus faecium por reação polimerase em cadeia. Foram analisadas 100 carcaças de frangos, das quais foram isoladas 50 cepas de Enterococcus spp., sendo 42% de E. faecalis e 2% de E. faecium. O teste de susceptibilidade antimicrobiana demonstrou que todas as cepas isoladas apresentaram resistência a pelo menos um antimicrobiano, dos quais 90,47% das cepas de E. faecalis, 100% das cepas de E. Faecium e 82,14% dos Enterococcus spp. apresentaram resistência à Tetraciclina; 80,95% das cepas de E. faecalis e 35,71% das cepas de Enterococcus spp. foram resistentes à Eritromicina; 39,28% dos Enterococcus spp. e 23,80% dos E. faecalis à Ciprofloxacina e 28,57% dos E. faecalis apresentaram resistência ao Cloranfenicol. Foram detectados os genes de resistência antimicrobiana erm(B, vanC-1, aph(3'-llla, ant(6-la, vanB, vanA, aac(6'-le-aph(2''-la, erm(A e tet(M - este último mais frequente. Estes resultados sugerem sérios problemas para a Saúde Pública, uma vez que esses microrganismos podem possuir a capacidade de transmitir genes de resistência antimicrobiana para outros microrganismos presentes na microbiota intestinal de humanos e animais, podendo inviabilizar o uso destas drogas para tratamentos clínicos.
Full Text Available Describimos el primer caso en Argentina de Staphylococcus aureus resistente a la meticilina adquirido en la comunidad con sensibilidad intermedia a la vancomicina y no sensibilidad a la daptomicina. Caso clínico: Paciente masculino, con antecedente de insuficiencia renal crónica en hemodiálisis y osteosíntesis de cadera debido a una fractura. Se internó por síndrome febril persistente luego del desplazamiento de la prótesis por un traumatismo. Se aisló de hemocultivos S. aureus resistente a la meticilina adquirido en la comunidad. Durante el tratamiento con vancomicina y daptomicina, se observó un aumento gradual de la CIM de vancomicina de 1 µg/ml (VSSA a 2 µg/ml (h-VISA y a 4 µg/ml (VISA, y la emergencia de no sensibilidad a daptomicina (CIM = 4 µg/ml. Al suspender la vancomicina y la daptomicina, la cepa revirtió al fenotipo de sensibilidad a ambas drogas. Es mandatorio evaluar por CIM la sensibilidad a vancomicina y a daptomicina intratratamiento cuando estas drogas se usan como terapia.
Full Text Available O objetivo deste estudo foi determinar o potencial probiótico in vitro de Lactobacillus spp. isolados de queijos minas artesanais da Serra da Canastra, considerando-se o antagonismo entre amostras isoladas frente a microrganismos indicadores, a susceptibilidade a antimicrobianos, a sensibilidade ao ácido gástrico e a sensibilidade a sais biliares. Todas as bactérias ácido-lácticas testadas apresentaram resistência ao ácido gástrico (pH 2,0 e aos sais biliares (0,3%, bem como atividade antagonista contra Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, Salmonella enterica var. Typhimurium, Enterococcus faecalis e bactérias ácido-lácticas isoladas dos próprios queijos - Lactobacillus plantarum (D27 e Lactobacillus rhamnosus (B25. Todas as amostras foram sensíveis à eritromicina e tetraciclina e resistentes à ciprofloxacina, gentamicina, oxacilina, estreptomicina e vancomicina. L. plantarum (B17 apresentou melhor potencial probiótico, pois obteve resultados satisfatórios em todas as propriedades avaliadas. Mais estudos são necessários para verificar a presença e a capacidade de transmissão de genes de resistência antimicrobiana a outros microrganismos e para avaliar o potencial dos microrganismos in vivo. As bactérias selecionadas poderão ser utilizadas na elaboração de queijos em que sejam mantidos o sabor e a tradição do queijo minas artesanal do estado de Minas Gerais.
Dahl, Anders; Rasmussen, Rasmus V; Bundgaard, Henning
Because of the nephrotoxic effects of aminoglycosides, the Danish guidelines on infective endocarditis were changed in January 2007, reducing gentamicin treatment in enterococcal infective endocarditis from 4 to 6 weeks to only 2 weeks. In this pilot study, we compare outcomes in patients...... with Enterococcus faecalis infective endocarditis treated in the years before and after endorsement of these new recommendations....
Dahl, Anders; Bruun, Niels Eske
Enterococcus faecalis infective endocarditis (IE) is a disease of increasing importance, with more patients infected, increasing frequency of health-care associated infections and increasing incidence of antimicrobial resistances. The typical clinical presentation is a subacute course with fever...... or ceftriaxone. E. faecalis infective endocarditis continues to be a very serious disease with considerable percentages of high-level gentamicin resistant strains and in-hospital mortality around 20%. Strategies to prevent E. faecalis IE, improve diagnostics, optimize treatment and reduce morbidity...
Rincón, Sandra; Panesso, Diana; Díaz, Lorena; Carvajal, Lina P.; Reyes, Jinnethe; Munita, José M.; Arias, César A.
En los últimos años se han desarrollado nuevas alternativas para el tratamiento de infecciones por patógenos Gram positivos multirresistentes, entre los cuales Staphylococcus aureus resistente a la meticilina (SARM) y los enterococos resistentes a la vancomicina (ERV) se consideran un verdadero reto terapéutico, y aunque el uso de la vancomicina en infecciones graves causadas por SARM ha generado serias dudas en los últimos años, continúa siendo escasa la información clínica de respaldo al uso de agentes terapéuticos que la superen en eficacia. El linezolid, la daptomicina y la tigeciclina son agentes que tienen actividad contra los cocos Gram positivos y que fueron aprobados e introducidos en la terapia clínica en la década pasada. Además, se han probado o están en las fases finales de desarrollo otros agentes como las cefalosporinas de última generación (ceftarolina y ceftobiprol). El propósito de esta revisión fue describir las nuevas alternativas terapéuticas, particularmente en la era posterior a la vancomicina, y repasar las características químicas más relevantes de los compuestos y su espectro de actividad, haciendo énfasis en sus mecanismos de acción y resistencia. PMID:24968051
Full Text Available En los últimos años se han desarrollado nuevas alternativas para el tratamiento de infecciones por patógenos Gram positivos multirresistentes, entre los cuales Staphylococcus aureus resistente a la meticilina (SARM y los enterococos resistentes a la vancomicina (ERV se consideran un verdadero reto terapéutico, y aunque el uso de la vancomicina en infecciones graves causadas por SARM ha generado serias dudas en los últimos años, continúa siendo escasa la información clínica de respaldo al uso de agentes terapéuticos que la superen en eficacia. El linezolid, la daptomicina y la tigeciclina son agentes que tienen actividad contra los cocos Gram positivos y que fueron aprobados e introducidos en la terapia clínica en la década pasada. Además, se han probado o están en las fases finales de desarrollo otros agentes como las cefalosporinas de última generación (ceftarolina y ceftobiprol. El propósito de esta revisión fue describir las nuevas alternativas terapéuticas, particularmente en la era posterior a la vancomicina, y repasar las características químicas más relevantes de los compuestos y su espectro de actividad, haciendo énfasis en sus mecanismos de acción y resistencia.
Leazer, Rianna; Perkins, Amy M; Shomaker, Kyrie; Fine, Bryan
A change in the epidemiology of pathogens causing serious bacterial infection (SBI) has been noted since original recommendations were made for the empirical antibiotic choices for young infants with fever. To assess the prevalence of SBI caused by Listeria monocytogenes and Enterococcus species. A literature search was conducted on keywords related to SBI, L. monocytogenes, and Enterococcus spp. infections. Eligible studies were those conducted in the United States and published between January 1998 and June 2014 focusing on SBI in infants≤90 days of age. The rates of urinary tract infection, bacteremia, and meningitis for each pathogen were recorded for each study. Meta-analysis was performed to calculate the prevalence for each pathogen in a random effects model with 0.5 continuity correction added to studies with zero events. Sixteen studies were included. A total of 20,703 blood cultures were included, with weighted prevalences for L. monocytogenes and Enterococcus spp. bacteremia of 0.03% and 0.09%, respectively. A total of 13,775 cerebrospinal fluid cultures were included with event rates (unweighted prevalences) for L. monocytogenes and Enterococcus spp. meningitis of 0.02% and 0.03%, respectively. A total of 18,283 urine cultures were included, with no cases of L. monocytogenes and a weighted prevalence for Enterococcus spp. urinary tract infection of 0.28%. There may have been reporting bias or incomplete retrieval or inadvertent exclusion of relevant studies. SBI caused by L. monocytogenes and Enterococcus spp. in febrile infants is rare, and therefore clinicians may consider a change in empirical antibiotic choices. Copyright © 2016 by the American Academy of Pediatrics.
Stępień-Pyśniak, Dagmara; Marek, Agnieszka; Banach, Tomasz; Adaszek, Łukasz; Pyzik, Ewelina; Wilczyński, Jarosław; Winiarczyk, Stanisław
The aim of this study was to evaluate the frequency of occurrence of bacteria of the genus Enterococcus in poultry, to identify them by means of matrixassisted laser desorption/ionisation time-of-flight mass spectrometry (MALDITOF MS), and to analyse the antimicrobial susceptibility of the isolated strains to the drugs most frequently used in poultry. The material for the bacteriological tests was obtained mainly from the heart (97%) of the birds investigated. Of a total of 2,970 samples tested, 911 (30.7%) tested positive for Enterococcus spp. Enterococci were detected in broilers (88.1%), laying hens (5.3%), turkeys (3.9%), breeding hens (2.2%), and geese (0.4%). The most commonly identified species were Enterococcus (E.) faecalis (74.7%), E. faecium (10.1%), E. gallinarum (5.5%), E. hirae (4.6%), and E. cecorum (4.1%). The most frequent resistance properties were resistance to sulphamethoxazole/trimethoprim (88%), tylosin (71.4%), enrofloxacin (69.4%), doxycycline (67.3%), and lincomycin/spectinomycin (56.1%). Only one vancomycin-resistant Enterococcus, E. cecorum from a broiler, was found.
Avcı, Mine; Özden Tuncer, Banu
The purpose of this study was to determine the antimicrobial activity and occurrence of bacteriocin structural genes in Enterococcus spp. isolated from different cheeses and also investigate some of their virulence factors. Enterococcus strains were isolated from 33 different cheeses. Enterococcus faecium (6 strains) and Enterococcus faecalis (5 strains) enterocin-producing strains were identified by 16S rDNA analyses. Structural genes entA, entB, entP and entX were detected in some isolates. Multiple enterocin structural genes were found in 7 strains. None of the tested enterococci demonstrated anyβ-haemolytic activity and only one strain had gelatinase activity. Six strains showed multiple antibiotic resistance patterns and in addition, vanA and several virulence genes were detected in many strains. Only E. faecalis MBE1-9 showed tyrosine decarboxylase activity and tdc gene was detected only in this strain.
Avaliação da tolerância à vancomicina em 395 cepas hospitalares de Staphylococcus aureus resistentes à oxacilina Evaluation of the tolerance to vancomycin in 395 oxacillin-resistant Staphylococcus aureus strains isolated from Brazilian hospitals
Geraldo A. Oliveira
Full Text Available O objetivo deste estudo foi avaliar a presença de tolerância à vancomicina em cepas de Staphylococcus aureus resistentes à oxacilina (Orsa isoladas de quatro hospitais da cidade de São Paulo. Foram estudadas 395 cepas Orsa isoladas de pacientes hospitalizados entre outubro de 1998 e maio de 2000. A determinação da concentração inibitória mínima (CIM e da concentração bactericida mínima (CBM para vancomicina foi realizada conforme padronizado pelo National Committee for Clinical Laboratory Standards (NCCLS. A tolerância à vancomicina foi definida como a razão CBM/CIM > ou = 32. Do total de cepas estudadas, 10,4% apresentaram CIM de 0,5µg/ml para vancomicina; 41,3%, CIM de 1µg/ml; 42,2%, CIM de 2µg/ml; e 6,1%, CIM de 4µg/ml. Em média, 49,1% dos Orsa apresentaram tolerância à vancomicina. Em conclusão, a tolerância à vancomicina entre as cepas Orsa foi considerada elevada. Conseqüentemente, aumentam as chances de falhas no tratamento com vancomicina, além de aumentar o risco da emergência de Staphylococcus aureus vancomicina-intermediário.The objective of this study was to evaluate the presence of tolerance to vancomycin in oxacillin-resistant Staphylococcus aureus (Orsa strains isolated from four hospitals in the city of São Paulo. From October/1998 to May/2000 we analysed 395 Orsa strains isolated from hospitalized patients. MIC and MBC to vancomycin were determined as standardised by National Committee for Clinical Laboratory Standards (NCCLS. Tolerance was defined as the ratio MBC/MIC > or = 32. The results showed that 10.4% of the Orsa strains presented a MIC of 0.5µg/mL for vancomycin, 41.3% presented a MIC of 1µg/mL, 42.2% a MIC of 2µg/mL and 6.1% a MIC of 4µg/mL. On average, 49,1% of the Orsa presented tolerance to vancomycin. We conclude that the tolerance to vancomycin amongst the Orsa strains was considered high. These high levels of tolerance augment the chances of failure in the treatment with
Doun, Stephanie S.; Burgner, John W.; Briggs, Scott D.; Rodwell, Victor W.
The six enzymes of the mevalonate pathway of isopentenyl diphosphate biosynthesis represent potential for addressing a pressing human health concern, the development of antibiotics against resistant strains of the Gram-positive streptococci. We previously characterized the first four of the mevalonate pathway enzymes of Enterococcus faecalis, and here characterize the fifth, phosphomevalonate kinase (E.C. 126.96.36.199). E. faecalis genomic DNA and the polymerase chain reaction were used to clone DNA thought to encode phosphomevalonate kinase into pET28b(+). Double-stranded DNA sequencing verified the sequence of the recombinant gene. The encoded N-terminal hexahistidine-tagged protein was expressed in Escherichia coli with induction by isopropylthiogalactoside and purified by Ni++ affinity chromatography, yield 20 mg protein per liter. Analysis of the purified protein by MALDI-TOF mass spectrometry established it as E. faecalis phosphomevalonate kinase. Analytical ultracentrifugation revealed that the kinase exists in solution primarily as a dimer. Assay for phosphomevalonate kinase activity used pyruvate kinase and lactate dehydrogenase to couple the formation of ADP to the oxidation of NADH. Optimal activity occurred at pH 8.0 and at 37°C. The activation energy was ~5.6 kcal/mol. Activity with Mn++, the preferred cation, was optimal at about 4 mM. Relative rates using different phosphoryl donors were 100 (ATP), 3.6 (GTP), 1.6 (TTP), and 0.4 (CTP). Km values were 0.17 mM for ATP and 0.19 mM for (R,S)-5-phosphomevalonate. The specific activity of the purified enzyme was 3.9 μmol substrate converted per minute per milligram protein. Applications to an immobilized enzyme bioreactor and to drug screening and design are discussed. PMID:15802646
Kilbas, Imdat; Ciftci, Ihsan Hakki
In this study, a meta-analysis of Enterococcus isolates collected in 2000-2015 in Turkey and their susceptibility/resistance to antibiotics, clinical indications for initial drug treatment, and identification of alternative treatments was conducted. The meta-analysis examined antibiotic susceptibility/resistance in Enterococcus spp. isolates. The study was planned and conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). Statements on antimicrobial resistance were grouped according to the antimicrobial stewardship programme (ASP). The mean resistance rates of Enterococcus faecalis to vancomycin (VAN) and linezolid (LNZ) were 1.0±2.2% and 1.9±2.6%, respectively, whereas the mean resistance rates of Enterococcus faecium to VAN and LNZ were 10.3±11.3% and 2.4±0%, respectively. This study is the first meta-analysis of the resistance of clinical Enterococcus isolates in Turkey to antimicrobial agents, which is a major problem stemming from the excessive usage of antibiotics. The development of antibiotic resistance in Turkey has changed over time. To support the practice of evidence-based medicine, more notifications about Enterococcus resistance status are needed, especially notifications following ASP rules. Copyright © 2017 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.
Full Text Available Objetivo: Estimar la costo-efectividad de linezolid versus vancomicina en el manejo de neumonía asociada a ventilación mecánica (NAV causada por Staphylococcus aureus resistente a meticilina (SARM en Colombia. Materiales y métodos: Se construyó un árbol de decisión para determinar la razón de costo-efectividad incremental de linezolid (600 mg iv/12 h comparado con vancomicina (15 mg/kg iv/12 h en el tratamiento de NAV por SARM. La perspectiva fue la del sistema de salud incluyendo solo costos directos. Todas las unidades monetarias se expresan en pesos colombianos del 2013 sin descuento (1 USD =$ 1.876,22. Se empleó un horizonte temporal de 30 días. Los resultados se midieron en proporción de pacientes curados. Los datos de eficacia y seguridad se tomaron de la literatura. Los costos de los procedimientos se obtuvieron del manual tarifario ISS del 2001, para medicamentos se utilizó el SISMED y la regulación de precios vigente. Se realizaron análisis de sensibilidad univariados y probabilísticos. Resultados: Los costos totales esperados por paciente curado fueron: $ 2.600.094 para linezolid y $ 1.992.753 para vancomicina. La proporción de pacientes curados fue: 53% con linezolid y 41%.con vancomicina. La razón de costo-efectividad de linezolid comparado con vancomicina fue $ 5.061.173 por paciente curado. Para cada alternativa, los resultados fueron sensibles a la probabilidad de éxito del tratamiento, a la probabilidad de presentar eventos adversos y al costo del tratamiento. Conclusión: En Colombia, linezolid sería una alternativa costo-efectiva en el tratamiento de NAV por SARM, para disponibilidades a pagar superiores a $ 5.061.173 por paciente curado.
Diana Alejandra Pico Veslin
Full Text Available Introducción: Los probióticos son conocidos como microrganismos vivos, que favorecen el equilibrio microbiano en el tracto gastrointestinal, sus metabolitos (ácidos orgánicos, ácido láctico, entre otros inhiben el crecimiento de patógenos, a nivel de sistema inmune activan la respuesta humoral (IL 2 y activan la fagocitosis. Diferentes estudios han demostrado que la utilización de probióticos en el sector avícola como suplemento, relegaría el uso de antibióticos, puesto que la utilización de los mismos, ha generado casos de resistencia bacteriana, afectando al consumidor final y convirtiéndose en un problema de salud pública. El uso de probióticos según la FAO, encaminaría en estrategias de mejoramiento global del rendimiento y bienestar animal en países en desarrollo. Objetivo: Identificar fenotípicamente el comportamiento de las cepas Enterorococcus faecium, Enterococcus faecalis aisladas del intestino de pollo de engorde y gallina comercial (Gallus gallus frente a antibacterianos. Metodología: estudio descriptivo de corte transversal. Se tomaron muestras del mucus intestinal de las aves (íleon, yeyuno y duodeno aisladas en medio MRS el cual permite evidenciar el crecimiento de bacterias ácido lácticas, se realizaron pruebas microbiológicas y bioquímicas utilizando el sistema semiautomatizado Crystal para Gram positivos (BBLTM CrystalTM Gram-PositiveID Kit posteriormente estas cepas fueron evaluadas en algunas características como potenciales probióticos: resistencia a pH bajo, concentraciones biliares de 0.1% a 0.3%, producción de hemolisinas, prueba de antagonismo; posteriormente se evaluó un factor de virulencia, la presencia de genes de resistencia a ciertos antibióticos; se realizó la caracterización fenotípica mediante difusión en disco Kirby Bauer , para evaluar la resistencia y/o sensibilidad, se seleccionaron los antibióticos de las familias Glicopetidos (Vancomicina, Inh
da Silva Fernandes, Meg; Kabuki, Dirce Yorika; Kuaye, Arnaldo Yoshiteru
The formation of mono-species biofilm (Listeria monocytogenes) and multi-species biofilms (Enterococcus faecium, Enterococcus faecalis, and L. monocytogenes) was evaluated. In addition, the effectiveness of sanitation procedures for the control of the multi-species biofilm also was evaluated. The biofilms were grown on stainless steel coupons at various incubation temperatures (7, 25 and 39°C) and contact times (0, 1, 2, 4, 6 and 8 days). In all tests, at 7°C, the microbial counts were below 0.4 log CFU/cm(2) and not characteristic of biofilms. In mono-species biofilm, the counts of L. monocytogenes after 8 days of contact were 4.1 and 2.8 log CFU/cm(2) at 25 and 39°C, respectively. In the multi-species biofilms, Enterococcus spp. were present at counts of 8 log CFU/cm(2) at 25 and 39°C after 8 days of contact. However, the L. monocytogenes in multi-species biofilms was significantly affected by the presence of Enterococcus spp. and by temperature. At 25°C, the growth of L. monocytogenes biofilms was favored in multi-species cultures, with counts above 6 log CFU/cm(2) after 8 days of contact. In contrast, at 39°C, a negative effect was observed for L. monocytogenes biofilm growth in mixed cultures, with a significant reduction in counts over time and values below 0.4 log CFU/cm(2) starting at day 4. Anionic tensioactive cleaning complemented with another procedure (acid cleaning, disinfection or acid cleaning+disinfection) eliminated the multi-species biofilms under all conditions tested (counts of all micro-organismsbiofilms under all tested conditions (counts of the all microorganisms biofilms under all the test conditions. Copyright © 2015 Elsevier B.V. All rights reserved.
Jahan, Musarrat; Zhanel, George G; Sparling, Richard; Holley, Richard A
Enterococcus species are part of the normal intestinal flora of a large number of mammals including humans and consequently, they can be used as indicators of faecal contamination in food and water for human consumption. Their presence in large numbers in foods may indicate a lapse in sanitation and their ability to serve as a genetic reservoir of transferable antibiotic resistance is of concern. In the present study, Enterococcus spp., isolated from commercially fermented meat and human clinical specimen were studied to determine genetic relationships. SmaI pulsed-field gel electrophoresis (PFGE) patterns exhibited genomic heterogeneity within and between both groups of isolates. However, in spite of this heterogeneity there were still substantial phenotypic similarities which suggested that food might be a potential vehicle for distribution of resistant bacteria among humans. In vitro conjugation experiments demonstrated transfer of the tetracycline resistant determinant, tet(M), from Enterococcus faecium S27 isolated from fermented sausage to clinical isolates of both E. faecium and Enterococcus faecalis. The streptomycin resistance of E. faecium S27 was also transferred to a clinical strain, E. faecalis 82916, which was confirmed by the presence of the streptomycin resistance gene, aadA, in the donor and transconjugant strains. Since the aadA gene is associated with a class 1 integron, results also suggested that resistance transfer might have occurred via an integron. It appears this is the first identification of a class 1 integron in E. faecium isolated from food. The importance of food enterococci as a reservoir of antibiotic resistance genes and the potential for their genetic transfer to human strains following consumption of uncooked or undercooked contaminated meat is underlined by this work. Copyright © 2015 Elsevier B.V. All rights reserved.
Vijayavel, K.; Byappanahalli, Muruleedhara N.; Whitman, Richard L.; Ebdon, J.; Taylor, H.; Kashian, D.R.
Bacteriophages are viruses living in bacteria that can be used as a tool to detect fecal contamination in surface waters around the world. However, the lack of a universal host strain makes them unsuitable for tracking fecal sources. We evaluated the suitability of two newly isolated Enterococcus host strains (ENT-49 and ENT-55) capable for identifying sewage contamination in impacted waters by targeting phages specific to these hosts. Both host strains were isolated from wastewater samples and identified as E. faecium by 16S rRNA gene sequencing. Occurrence of Enterococcus phages was evaluated in sewage samples (n = 15) from five wastewater treatment plants and in fecal samples from twenty-two species of wild and domesticated animals (individual samples; n = 22). Levels of Enterococcus phages, F + coliphages, Escherichia coli and enterococci were examined from four rivers, four beaches, and three harbors. Enterococcus phages enumeration was at similar levels (Mean = 6.72 Log PFU/100 mL) to F + coliphages in all wastewater samples, but were absent from all non-human fecal sources tested. The phages infecting Enterococcus spp. and F + coliphages were not detected in the river samples (detection threshold < 10 PFU/100 mL), but were present in the beach and harbor samples (range = 1.83 to 2.86 Log PFU/100 mL). Slightly higher concentrations (range = 3.22 to 3.69 Log MPN/100 mL) of E. coli and enterococci when compared to F + coliphages and Enterococcus phages, were observed in the river, beach and harbor samples. Our findings suggest that the bacteriophages associated with these particular Enterococcus host strains offer potentially sensitive and human-source specific indicators of enteric pathogen risk.
Maasjost, J; Mühldorfer, K; Cortez de Jäckel S; Hafez, H M
Between 2010 and 2011, 145 Enterococcus isolates (Enterococcus faecalis, n = 127; Enterococcus faecium, n = 18) were collected during routine bacteriologic diagnostics from broilers, layers, and fattening turkeys in Germany showing various clinical signs. The susceptibility to 24 antimicrobial agents was investigated by broth microdilution test to determine minimum inhibitory concentrations (MICs). All E. faecalis isolates (n = 127) were susceptible to the beta-lactam antibiotics ampicillin, amoxicillin-clavulanic acid, and penicillin. Corresponding MIC with 50% inhibition (MIC50) and MIC with 90% inhibition (MIC90) values of these antimicrobial agents were at the lower end of the test range (≤ 4 μg/ml). In addition, no vancomycin-resistant enterococci (VRE) were found. High resistance rates were identified in both Enterococcus species for lincomycin (72%-99%) and tetracycline (67%-82%). Half or more than half of Enterococcus isolates were resistant to gentamicin (54%-72%) and the macrolide antibiotics erythromycin (44%-61%) and tylosin-tartate (44%-56%). Enterococcus faecalis isolated from fattening turkeys showed the highest prevalence of antimicrobial resistance compared to other poultry production systems. Eighty-nine out of 145 Enterococcus isolates were resistant to three or more antimicrobial classes. Again, turkeys stood out with 42 (8 1%) multiresistant isolates. The most-frequent resistance patterns of E. faecalis were gentamicin, lincomycin, and tetracycline in all poultry production systems.
Kjerulf, A; Espersen, F; Gutschik, E
A modified rat model of endocarditis with catheterization for 2 days was established in female Lewis rats using different inocula of Enterococcus faecalis (strain no. EF 19) in order to measure IgG antibodies in serum during the course of infection. Increasing the inocula intravenously resulted...
In 1903 the genus name Enterococcus was proposed for gram-positive, catalase-negative, coccoid-shaped bacterial of intestinal origin. Several years later, it was suggested that the genus name be changed to Streptococcus because of the organisms' ability to form chains of coccoid...
No Brasil, o papagaio-verdadeiro (Amazona aestiva) é uma das aves mais procuradas como animal de estimação e comercializadas ilegalmente. Na literatura pouco é descrito sobre a microbiota intestinal de aves silvestres. O trato intestinal das aves é composto por inúmeras e diferentes espécies bacterianas. A grande maioria são bactérias gram-positivas pertencentes ao grupo de bactérias ácido-láticas. Este estudo teve como objetivo isolar e identificar a presença de bactérias dos gêneros Lactoba...
Carolina Baldisserotto Comerlato
Full Text Available Despite the increasing importance of Enterococcus as opportunistic pathogens, their virulence factors are still poorly understood. This study determines the frequency of virulence factors in clinical and commensal Enterococcus isolates from inpatients in Porto Alegre, Brazil. Fifty Enterococcus isolates were analysed and the presence of the gelE, asa1 and esp genes was determined. Gelatinase activity and biofilm formation were also tested. The clonal relationships among the isolates were evaluated using pulsed-field gel electrophoresis. The asa1, gelE and esp genes were identified in 38%, 60% and 76% of all isolates, respectively. The first two genes were more prevalent in Enterococcus faecalis than in Enterococcus faecium, as was biofilm formation, which was associated with gelE and asa1 genes, but not with the esp gene. The presence of gelE and the activity of gelatinase were not fully concordant. No relationship was observed among any virulence factors and specific subclones of E. faecalis or E. faecium resistant to vancomycin. In conclusion, E. faecalis and E. faecium isolates showed significantly different patterns of virulence determinants. Neither the source of isolation nor the clonal relationship or vancomycin resistance influenced their distribution.
Soheili, Sara; Ghafourian, Sobhan; Sekawi, Zamberi; Neela, Vasanthakumari; Sadeghifard, Nourkhoda; Ramli, Ramliza; Hamat, Rukman Awang
Enterococcus, a Gram-positive facultative anaerobic cocci belonging to the lactic acid bacteria of the phylum Firmicutes, is known to be able to resist a wide range of hostile conditions such as different pH levels, high concentration of NaCl (6.5%), and the extended temperatures between 5(°)C and 65(°)C. Despite being the third most common nosocomial pathogen, our understanding on its virulence factors is still poorly understood. The current study was aimed to determine the prevalence of different virulence genes in Enterococcus faecalis and Enterococcus faecium. For this purpose, 79 clinical isolates of Malaysian enterococci were evaluated for the presence of virulence genes. pilB, fms8, efaAfm, and sgrA genes are prevalent in all clinical isolates. In conclusion, the pathogenicity of E. faecalis and E. faecium could be associated with different virulence factors and these genes are widely distributed among the enterococcal species.
Full Text Available Background and Objectives: Enterococci are Gram-positive cocci that are found in abundance in the environment. The ability of these bacteria for long-term survival in hospital environment, increases the chance of patients for infections caused by this bacterium. Therefore, identification of possible sources and reservoirs is helpful in identifying the potential sources in sudden outbreaks. The current study was conducted to determine the prevalence rate and antibiotic resistance pattern in environmental Enterococcus isolates collected from delivery room environment of Alzahra and Izadi university hospitals in Qom. Methods: In this descriptive cross-sectional study, over a period of 3 month, sampling was performed using swabbing method from delivery rooms of alzahra and izadi hospitals in Qom, and were examined for the presence of Enterococci. After isolation, Enterococcus species were examined using different biochemical tests, and the antibiotic resistance pattern in the environmental was assessed according to disk-diffusion test according to Clinical and Laboratory Standards Institute (CLSI standards. The data were described by frequency tables. Results: In this study, out of 216 obtained isolates, 7(3.2% belonged to Enterococcus genus. 4(57% out of 7 Enterococcus strains isolated from surfaces of delivery room, had multiple resistances based on antibiotic sensitivity test using antiobiogram test. Conclusion: According to increasing growth of nosocomial infection caused by resistant strains of Enterococcus spp. and their spread in hospital environment, use of infection control measures are necessary to eliminate the potential sources and prevent the infection.
Kim, Y-J; Park, J-H; Seo, K-H
Antibiotic-resistant bacteria in poultry meat are a threat to public health. In this study, we compared the Enterococcus spp. loads and antibiotic-resistance profiles between carcasses of conventionally and organically raised chickens. A total of 144 chicken carcasses (72 conventional and 72 organic) was collected from local retail markets in Seoul, South Korea. Overall, 77.7% (112 of 144; 75% conventional and 80% organic) of chicken carcasses were positive for Enterococcus. The mean loads of Enterococcus spp. were greater in conventional chicken carcasses, at 2.9 ± 0.4 log CFU/mL, than those in organic chicken carcasses, at 1.78 ± 0.3 log CFU/mL (p organic chicken carcasses) was randomly selected for further analysis. The predominant species was Enterococcus faecalis in both conventional and organic chicken carcasses (57.7 and 76.9%, respectively; P > 0.05). Rates of resistance to ciprofloxacin and erythromycin, which are used in veterinary medicine in South Korea, were significantly higher in conventional chicken carcasses than in organic chicken carcasses. However, we found no difference between the rates of resistance to antibiotics such as vancomycin and tigecycline, which were not registered for use in veterinary medicine in South Korea, of Enterococcus isolates from conventional and organic chicken carcasses. In addition, although multidrug resistant isolates were obtained from both types of chicken samples, the prevalence of samples positive for Enterococcus was significantly higher in conventional chicken carcasses than in organic chicken carcasses (P organic chicken carcasses. A high level of gentamicin resistance was observed in isolates from not only conventional (5.8%) but also organic chicken (1.9%) carcasses, with no significant difference in rates between them (P > 0.05). Despite this, our results suggest that organic food certification is effective in reducing fecal contamination and the burden of antibiotic-resistant Enterococcus spp. in
Mikalsen, Theresa; Pedersen, Torunn; Willems, Rob; Coque, Teresa M; Werner, Guido; Sadowy, Ewa; van Schaik, Willem; Jensen, Lars Bogø; Sundsfjord, Arnfinn; Hegstad, Kristin
BACKGROUND: The success of Enterococcus faecium and E. faecalis evolving as multi-resistant nosocomial pathogens is associated with their ability to acquire and share adaptive traits, including antimicrobial resistance genes encoded by mobile genetic elements (MGEs). Here, we investigate this
Mikalsen, Theresa; Pedersen, Torunn; Willems, Rob
Background: The success of Enterococcus faecium and E. faecalis evolving as multi-resistant nosocomial pathogens is associated with their ability to acquire and share adaptive traits, including antimicrobial resistance genes encoded by mobile genetic elements (MGEs). Here, we investigate this mob...
Bal, Emel Banu Buyukunal; Isevi, Taner; Bal, Mehmet Ali
Two Enterococcus faecium and one E. faecalis strains isolated and identified from wheat silage were characterized based on plasmid content, hemolytic activity, antibiotic resistance patterns, bacteriocin production potential, and presence of enterocin structural genes (entA, entB, entP, entL50B). Among the isolates, only the E. faecium U7 strain exhibited bacteriocin activity against Listeria monocytogenes ATCC 7644, and vancomycin resistant Enterococcus spp. (VRE). A combination of three structural genes (entA, entB, and entP) was detected in E. faecium U7. A relationship between the presence of enterocin structural genes, and bacteriocin activity was detected in E. faecium U7; therefore partially purified enterocin (PPE) was further investigated from the isolate. Several bands of different molecular weights were expressed from PPE extracts following tricine SDS-PAGE analysis. However, the only band showing bacteriocin activity was in an approximate 4-kDa region. PPE treatment with proteinase K, lysozyme, and α -amylase caused complete loss of bacteriocin activity. PPE heat treatment at various temperatures resulted in a notable reduction in bacteriocin expression. Enterocin U7 was relatively heat stable, and presumably exhibits a glucoprotein nature with distinct inhibitory properties. Specific bacterial inhibitory activity of enterocin U7, and the producer strain absence of β -hemolysis and vancomycin susceptibility features deserves further investigation to evaluate its potential application in silage inoculation and food preservation. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Fang, Xiang-Qun; Liu, You-Ning
to study the diagnosis and treatment of enterococcus faecium lung abscess. a retrospective analysis of one case of Enterococcus faecium lung abscess and literature review was conducted. this patient suffered from cough and sputum over 6 months and complicated with hemoptysis over 3 months. Pulmonary embolism and lung cancer were suspected initially. After 2 times of CT-guided percutaneous transthoracic needle aspiration biopsy the diagnosis of pneumonia was made in other hospitals. However, the consolidation in the lung progressed and cavity appeared although antibiotic therapy was conducted. After admission to our hospital, CT-guided percutaneous transthoracic needle aspiration biopsy was made and the lung tissue was sent for bacterial culture. Enterococcus faecium was cultured and it was susceptible to vancomycin, teicoplanin and linezolid. The disease improved significantly after treatment with these 3 antibiotics in turn. In addition, 13 cases of enterococcus pneumonia or lung abscess were reviewed, including 3 cases of enterococcus faecium lung abscess. enterococcus faecium is rarely a pathogen for lung abscess. The diagnosis of enterococcus faecium lung abscess could be confirmed by lung biopsy and bacterial culture of lung tissue which could also provide the susceptibility of antibiotics and guide the antibiotic therapy.
Full Text Available Enterococcus, a Gram-positive facultative anaerobic cocci belonging to the lactic acid bacteria of the phylum Firmicutes, is known to be able to resist a wide range of hostile conditions such as different pH levels, high concentration of NaCl (6.5%, and the extended temperatures between 5°C and 65°C. Despite being the third most common nosocomial pathogen, our understanding on its virulence factors is still poorly understood. The current study was aimed to determine the prevalence of different virulence genes in Enterococcus faecalis and Enterococcus faecium. For this purpose, 79 clinical isolates of Malaysian enterococci were evaluated for the presence of virulence genes. pilB, fms8, efaAfm, and sgrA genes are prevalent in all clinical isolates. In conclusion, the pathogenicity of E. faecalis and E. faecium could be associated with different virulence factors and these genes are widely distributed among the enterococcal species.
Full Text Available Abstract Background Iron-sulfur clusters are ubiquitous and evolutionarily ancient inorganic prosthetic groups, the biosynthesis of which depends on complex protein machineries. Three distinct assembly systems involved in the maturation of cellular Fe-S proteins have been determined, designated the NIF, ISC and SUF systems. Although well described in several organisms, these machineries are poorly understood in Gram-positive bacteria. Within the Firmicutes phylum, the Enterococcus spp. genus have recently assumed importance in clinical microbiology being considered as emerging pathogens for humans, wherein Enterococcus faecalis represents the major species associated with nosocomial infections. The aim of this study was to carry out a phylogenetic analysis in Enterococcus faecalis V583 and a structural and conformational characterisation of it SufU protein. Results BLAST searches of the Enterococcus genome revealed a series of genes with sequence similarity to the Escherichia coli SUF machinery of [Fe-S] cluster biosynthesis, namely sufB, sufC, sufD and SufS. In addition, the E. coli IscU ortholog SufU was found to be the scaffold protein of Enterococcus spp., containing all features considered essential for its biological activity, including conserved amino acid residues involved in substrate and/or co-factor binding (Cys50,76,138 and Asp52 and, phylogenetic analyses showed a close relationship with orthologues from other Gram-positive bacteria. Molecular dynamics for structural determinations and molecular modeling using E. faecalis SufU primary sequence protein over the PDB:1su0 crystallographic model from Streptococcus pyogenes were carried out with a subsequent 50 ns molecular dynamic trajectory. This presented a stable model, showing secondary structure modifications near the active site and conserved cysteine residues. Molecular modeling using Haemophilus influenzae IscU primary sequence over the PDB:1su0 crystal followed by a MD
Jan 10, 2012 ... inhibition of Enterococcus faecium 410 CECT in 6 h of incubation. The highest ... as natural food additives for the elimination of spoilage and pathogeinic ..... autohydrolysed fish viscera for nisin and pediocin production. J.
S. Hernández Egido
Full Text Available Small colony variants (SCV are slow-growing subpopulations of bacteria usually associated with auxotrophism, causing persistent or recurrent infections. Enterococcus faecalis SCV have been seldom described, and only one case of Enterococcus faecium SCV has been reported, associated with sepsis in a leukaemia patient. Here we report the first case described of bacteraemia and endocarditis by SCV E. faecium in an immunocompetent patient.
Sulzer, G; Busse, M
Bacterial strains exhibiting antimicrobial activity towards other bacteria are quite common in nature. During the past few years several genera have been shown to exert inhibitory action against Listeria. spp. In the present work strains of Enterococcus, Lactobacillus and Lactococcus were tested for their influence on the development of Listeria spp. on Camembert cheese. Partial or complete inhibition of growth of Listeria spp. was observed using various inhibitory bacteria. Complete inhibition occurred when the inhibitory strain was used as a starter culture and there was a low level of contamination with Listeria spp. during the first stage of ripening. Very little inhibition occurred if the inhibitory strain was added together with the starter culture.
Jul 31, 2012 ... Intermittent assessment of resistance genes in the ecosystem should be ..... among resistant Acinetobacter spp. isolated from integrated fish .... independent studies on the emerging phylogenetic view of bacterial .... Functional.
Cook, Laura C.; Federle, Michael J.
Intercellular chemical signaling in bacteria, commonly referred to as quorum sensing (QS), relies on the production and detection of compounds known as pheromones to elicit coordinated responses among members of a community. Pheromones produced by Gram-positive bacteria are comprised of small peptides. Based on both peptide structure and sensory system architectures, Gram-positive bacterial signaling pathways may be classified into one of four groups with a defining hallmark: cyclical peptides of the Agr type, peptides that contain Gly-Gly processing motifs, sensory systems of the RNPP family, or the recently characterized Rgg-like regulatory family. The recent discovery that Rgg family members respond to peptide pheromones increases substantially the number of species in which QS is likely a key regulatory component. These pathways control a variety of fundamental behaviors including conjugation, natural competence for transformation, biofilm development, and virulence factor regulation. Overlapping QS pathways found in multiple species and pathways that utilize conserved peptide pheromones provide opportunities for interspecies communication. Here we review pheromone signaling identified in the genera Enterococcus and Streptococcus, providing examples of all four types of pathways. PMID:24118108
Full Text Available We report a case of a 74-year-old female, with a mitral heart valve, who presented with pain and blurred vision in the right eye for 2 days. Her visual acuity was light perception (LP in the right eye and 20/40 in the left eye. Slit lamp examination showed corneal edema and hypopyon, and a view of the right fundus was impossible. Echography showed vitreous condensation. One day after presentation, the patient developed acute lung edema requiring hospitalization, so she was not submitted to vitreous tap and intravitreal treatment. The cardiac and systemic evaluations revealed a mitral endocarditis secondary to Enterococcus faecalis. The patient improved systemically with treatment with gentamicin, vancomycin, and linezolid. Her visual acuity remained as no LP, and her intraocular pressure (IOP has been controlled with brimonidine bid despite developing a total cataract with 360° posterior synechia. A cardiac source for endogenous endophthalmitis should be considered in the presence of a prosthetic cardiac valve. The treatment and followup must be made in cooperation with a cardiologist specialist, but the ophthalmologist can play a key role in the diagnosis.
Avaliação da sensibilidade a antimicrobianos de 87 amostras clínicas de enterococos resistentes à vancomicina Antimicrobial susceptibility testing of 87 clinical isolates of vancomycin-resistant enterococci
Full Text Available OBJETIVOS. 1 Avaliar o padrão de sensibilidade in vitro de amostras clínicas de enterococos resistentes à vancomicina (ERV, aos antimicrobianos comumente utilizados no seu tratamento, bem como a antimicrobianos alternativos. 2 Avaliar a acurácia do E test, em comparação aos outros testes de sensibilidade a antimicrobianos (microdiluição em caldo e difusão em disco. MATERIAL E MÉTODOS. Foram analisadas 87 amostras clínicas de ERVs selecionadas de 1.936 isolados de enterococos coletados em 97 hospitais norte-americanos, no último trimestre de 1992. A identificação em nível de espécie foi feita pelos sistemas API 20S, Vitek e uma versão modificada do método convencional proposto por Facklam e Collins. A avaliação da sensibilidade in vitro aos antimicrobianos foi realizada pela técnica de microdiluição em caldo, E test e métodos de difusão em disco. As amostras foram testadas, tanto para antimicrobianos normalmente utilizados no tratamento de infecções enterocócicas (vancomicina, teicoplanina, ampicilina, penicilina, gentamicina e estreptomicina, como também para drogas alternativas potencialmente úteis (cloranfenicol, doxiciclina, esparfloxacina, ciprofloxacina, clinafloxacina, eritromicina, espectinomicina, trospectomicina, trimetoprim-sulfametoxazol e novobiocina. RESULTADOS. A avaliação dos testes de sensibilidade das 87 amostras de ERV revelou resistência, a ampicilina em torno de 86%, o mesmo sendo observado para penicilina. Em relação aos aminoglicosídeos, obtivemos alto grau de resistência, em torno de 82% e 85%, para gentamicina e estreptomicina, respectivamente. Apesar de pertencer à mesma classe da vancomicina, a teicoplanina foi ativa contra 29% das amostras de ERV. Entre os antimicrobianos alternativos testados, os que apresentaram maiores taxas de sensibilidade foram o cloranfenicol, a doxiciclina e a trospectomicina (82%, 92% e 94% de isolados suscetíveis, respectivamente. CONCLUSÃO. O
Łysakowska, Monika E; Sienkiewicz, Monika; Banaszek, Katarzyna; Sokołowski, Jerzy
Enterococci are able to survive endodontic procedures and contribute to the failure of endodontic therapy. Thus, it is essential to identify novel ways of eradicating them from infected root canals. One such approach may be the use of antimicrobials such as plant essential oils. Enterococcal strains were isolated from endodontically treated teeth by standard microbiological methods. Susceptibility to antibiotics was evaluated by the disc-diffusion method. The minimal inhibitory concentration (MIC) of geranium essential oil was investigated by microdilution in 96-well microplates in Mueller Hinton Broth II. Biofilm eradication concentrations were checked in dentin tests. Geranium essential oil inhibited enterococcal strains at concentrations ranging from 1.8-4.5 mg/mL. No correlation was shown between resistance to antibiotics and the MICs of the test antimicrobials. The MICs of the test oil were lower than those found to show cytotoxic effects on the HMEC-1 cell line. Geranium essential oil eradicated enterococcal biofilm at concentrations of 150 mg/mL. Geranium essential oil inhibits the growth of endodontic enterococcal species at lower concentrations than those required to reach IC50 against the HMEC-1 cell line, and is effective against bacteria protected in biofilm at higher concentrations. In addition, bacteria do not develop resistance to essential oils. Hence, geranium essential oil represents a possible alternative to other antimicrobials during endodontic procedures.
Białucha, Agata; Kozuszko, Sylwia; Gospodarek, Eugenia; Bugalski, Roman Marian; Gierlotka, Krzysztof
Bacteriocins are ribosomally synthesised, extracellular bacterial products. Generally, spectrum of inhibition is limited to the same or closely related species to bacteriocin producer. Staphylococcin T is produced by Staphylococcus cohnii strain. The present study concerns influence of StT to 267 Enterococcus sp. strains growth isolated between 2003 and 2006 in Department of Microbiology University Hospital of dr. A. Jurasz in Bydgoszcz. S. cohnii T antagonistic ability evaluated towards bacteries on Mueller-Hinton Agar (bio Mérieux) in aerobic conditions. After 24 and 48 hours tested enterococci suspensions were plated perpendiculary. Susceptibility to antibiotics was assessed by disc diffusion method according to the guideless of Clinical and Laboratory Standards Institute and National Reference Centre for Antimicrobial Susceptibility. Among Enterococcus sp. strains tested 7.1% were sensitive to StT. The highest percentage of sensitive enterococci isolated from wound swabs, urine, blood and pus. Enterococcus faecium strains dominated (63.2%) among enterococci sensitive to StT. Moderate inhibition degree on S. cohnii T bacteriocin action was observed in majority sensitive enterococci strains. Enterococcus sp. sensitive to StT strains were frequently multidrug resistant (68.4%). According to the study results and increasing resistance to antibiotics, StT could be an alternative agent used to treat infections caused by Enterococcus sp.
Nilay Sefa Uçar
Full Text Available Enterococci are present in the gastrointestinal system as normal floral components. In the past two decades membersof the genus Enterococcus have emerged as important nosocomial pathogens worldwide. Enterococci may cause arange of different disorders such as urinary tract, intraabdominal, and wound infections, as well as endocarditis, meningitisand bacteraemia. Nosocomial enterococcal meningitis is most commonly observed following ventriculoperitonealshunt operations. Vancomycin resistant enterococcus (VRE represents 30% of all enterococci infections.This report presents a vancomycin-resistant Enterococcus casseliflavus meningitis case in a 66-year-old patient withventriculoperitoneal shunt, which has not been reported in the literature before. Successful outcomes were obtainedwith daptomycin plus linezolid combined treatment in VRE meningitis. Treatment recommendations in VRE meningitisare also discussed in this article. J Microbiol Infect Dis 2011;1 (3:138-140
Yuen, Grace J; Ausubel, Frederick M
The enterococci are commensals of the gastrointestinal tract of many metazoans, from insects to humans. While they normally do not cause disease in the intestine, they can become pathogenic when they infect sites outside of the gut. Recently, the enterococci have become important nosocomial pathogens, with the majority of human enterococcal infections caused by two species, Enterococcus faecalis and Enterococcus faecium. Studies using invertebrate infection models have revealed insights into the biology of enterococcal infections, as well as general principles underlying host innate immune defense. This review highlights recent findings on Enterococcus infection biology from two invertebrate infection models, the greater wax moth Galleria mellonella and the free-living bacteriovorous nematode Caenorhabditis elegans.
Rathnayake, I U; Hargreaves, M; Huygens, F
This study compared virulence and antibiotic resistance traits in clinical and environmental Enterococcus faecalis and Enterococcus faecium isolates. E. faecalis isolates harboured a broader spectrum of virulence determinants compared to E. faecium isolates. The virulence traits Cyl-A, Cyl-B, Cyl-M, gel-E, esp and acm were tested and environmental isolates predominantly harboured gel-E (80% of E. faecalis and 31.9% of E. faecium) whereas esp was more prevalent in clinical isolates (67.8% of E. faecalis and 70.4% of E. faecium). E. faecalis and E. faecium isolated from water had different antibiotic resistance patterns compared to those isolated from clinical samples. Linezolid resistance was not observed in any isolates tested and vancomycin resistance was observed only in clinical isolates. Resistance to other antibiotics (tetracycline, gentamicin, ciprofloxacin and ampicillin) was detected in both clinical and water isolates. Clinical isolates were more resistant to all the antibiotics tested compared to water isolates. Multi-drug resistance was more prevalent in clinical isolates (71.2% of E. faecalis and 70.3% of E. faecium) compared to water isolates (only 5.7% E. faecium). tet L and tet M genes were predominantly identified in tetracycline-resistant isolates. All water and clinical isolates resistant to ciprofloxacin and ampicillin contained mutations in the gyrA, parC and pbp5 genes. A significant correlation was found between the presence of virulence determinants and antibiotic resistance in all the isolates tested in this study (pantibiotic resistant enterococci, together with associated virulence traits, in surface recreational water could be a public health risk. Copyright © 2012 Elsevier GmbH. All rights reserved.
Benson Chuks Iweriebor
Full Text Available Background: Antimicrobial resistance in microorganisms are on the increase worldwide and are responsible for substantial cases of therapeutic failures. Resistance of species of Enterococcus to antibiotics is linked to their ability to acquire and disseminate antimicrobial resistance determinants in nature, and wastewater treatment plants (WWTPs are considered to be one of the main reservoirs of such antibiotic resistant bacteria. We therefore determined the antimicrobial resistance and virulence profiles of some common Enterococcus spp that are known to be associated with human infections that were recovered from hospital wastewater and final effluent of the receiving wastewater treatment plant in Alice, Eastern Cape. Methods: Wastewater samples were simultaneously collected from two sites (Victoria hospital and final effluents of a municipal WWTP in Alice at about one to two weeks interval during the months of July and August 2014. Samples were screened for the isolation of enterococci using standard microbiological methods. The isolates were profiled molecularly after targeted generic identification and speciation for the presence of virulence and antibiotic resistance genes. Results: Out of 66 presumptive isolates, 62 were confirmed to belong to the Enterococcus genusof which 30 were identified to be E. faecalis and 15 E. durans. The remaining isolates were not identified by the primers used in the screening procedure. Out of the six virulence genes that were targeted only three of them; ace, efaA, and gelE were detected. There was a very high phenotypic multiple resistance among the isolates and these were confirmed by genetic analyses. Conclusions: Analyses of the results obtained indicated that hospital wastewater may be one of the sources of antibiotic resistant bacteria to the receiving WWTP. Also, findings revealed that the final effluent discharged into the environment was contaminated with multi-resistant enterococci species thus
Imad AL Kassaa
Full Text Available AL Kassaa, I., Safourim, N., Mostafa, N. and Hamze, M. Antimicrobial activity and high thermostability of a novel BLIS secreted by Enterococcus Mundtii isolated from lebanese cow’s milk. 2016. Lebanese Science Journal, 17(2: 166-176. Lactic acid bacteria (LAB are used in many fields such as fermentation agents, increasing nutritional value and improving organoleptic quality of food. Also they are used as probiotics and preservatives against pathogens and spoilage microbes by producing antimicrobial substances such as bacteriocins. Fifty cow’s milk samples were collected and 175 LAB isolates were isolated and identified by using biochemical method. Fifteen isolates showed an antimicrobial activity against Listeria monocytogenes ATCC® 19115™. One strain, BL4 which showed the strongest activity, was chosen to extract and characterize its antimicrobial substance in order to evaluate its potential use as a new food protective agent. This strain was identified as Enterococcus mundtii by pyrosequencing method. The active substance was extracted using solvent method. This Bacteriocin like Inhibitory Substances “BLIS” can support a high temperature (121 ˚C for a long time and resist pH variation. The BLIS BL4 can be considered as a peptide active against many food pathogen and food-spoilage microbes, such as Listeria monocytogenes and Penicillium spp. BLIS BL4 can be used in food application as bio-preservative to reduce food-spoilage and food-borne diseases in food products.
Christ, Ana Paula Guarnieri; Ramos, Solange Rodrigues; Cayô, Rodrigo; Gales, Ana Cristina; Hachich, Elayse Maria; Sato, Maria Inês Zanoli
MALDI-TOF Mass Spectrometry Biotyping has proven to be a reliable method for identifying bacteria at the species level based on the analysis of the ribosomal proteins mass fingerprint. We evaluate the usefulness of this method to identify Enterococcus species isolated from marine recreational water at Brazilian beaches. A total of 127 Enterococcus spp. isolates were identified to species level by bioMérieux's API® 20 Strep and MALDI-TOF systems. The biochemical test identified 117/127 isolates (92%), whereas MALDI identified 100% of the isolates, with an agreement of 63% between the methods. The 16S rRNA gene sequencing of isolates with discrepant results showed that MALDI-TOF and API® correctly identified 74% and 11% of these isolates, respectively. This discrepancy probably relies on the bias of the API® has to identify clinical isolates. MALDI-TOF proved to be a feasible approach for identifying Enterococcus from environmental matrices increasing the rapidness and accuracy of results. Copyright © 2017 Elsevier Ltd. All rights reserved.
Primary acute conununity-acquired pneumonia was unconunon. ... organisms such as Klebsiella spp. are frequent causes of nosocomial infections."o The larrer ... Accepted 11 Sep 1992. recognised cause of community-acquired Gram-nega-.
Deng, Dong Mei; Hoogenkamp, Michel A.; Exterkate, Rob A. M.; Jiang, Lei Meng; van der Sluis, Lucas W. M.; ten Cate, Jacob M.; Crielaard, Wim
Introduction: An important virulence factor of Enterococcus faecalis is its ability to form biofilms. Most studies on biofilm formation have been carried out by using E. faecalis monocultures. Given the polymicrobial nature of root canal infections, it is important to understand biofilm formation of
Bacteriocin-like substance E204 is an antimicrobial compound produced by Enterococcus durans E204 isolated from camel milk of Morocco that shows a broad spectrum of inhibitory activity against taxonomically related microorganisms. It is sensitive to digestive proteases. In the first study, de Man, Regosa and Sharpe ...
Dahl, Anders; Lauridsen, Trine K; Arpi, Magnus
BACKGROUND: The NOVA score is a recently developed diagnostic tool to identify patients with increased risk of infective endocarditis (IE) among patients with Enterococcus faecalis (EF) bacteremia. We aim to validate an adapted version of the NOVA score and to identify risk factors for IE...
Jahan, M; Holley, R A
Listeria monocytogenes is an important foodborne pathogen that can cause infection in children, pregnant women, the immunocompromised and the elderly. Antibiotic resistance in this species would represent a significant public health problem since the organism has a high fatality/case ratio and resistance may contribute to failure of therapeutic treatment. This study was designed to explore whether the in vitro transferability of antibiotic resistance from enterococci to Listeria spp. could occur. It was found that 2/8 Listeria strains were able to acquire tetracycline resistance from Enterococcus faecium. Listeria monocytogenes GLM-2 acquired the resistance determinant tet(M) and additional streptomycin resistance through in vitro mating with Ent. faecium S27 isolated from commercial fermented dry sausage. Similarly, Listeria innocua became more resistant to tetracycline, but the genetic basis for this change was not confirmed. It has been suggested that enterococci may transfer antibiotic resistance genes via transposons to Listeria spp., and this may explain, in part, the origin of their antibiotic resistance. Thus, the presence of enterococci in food should not be ignored since they may actively contribute to enhanced antibiotic resistance of L. monocytogenes and other pathogens. Acquisition of antibiotic resistance by pathogenic bacteria in the absence of antibiotic pressure represents an unquantified threat to human health. In the present work resistance to tetracycline and streptomycin were transferred by nonplasmid-based conjugation from Enterococcus faecium isolated from fermented sausage to Listeria monocytogenes and Listeria innocua. Thus, natural transfer of antibiotic resistance to Listeria strains may occur in the future which reinforces the concern about the safety of enterococcal strains present in foods. © 2016 The Society for Applied Microbiology.
Full Text Available Enterococcus faecium EM41 is an isolate from ostrich faeces. It produces a thermo-stable proteinaceous substance, bacteriocin (enterocin EM41 with the highest inhibition activity in late logarithmic phase of growth (25 600 AU/ml. This strain and its enterocin have not been previously tested in animals. Lohmann Brown laying hens (aged 45 weeks were involved in this model/pilot experiment, divided into 3 groups 6 birds in each. E. faecium EM41 applied was a variant treated with rifampicin (109 cfu/ml, dose 400 μl/animal/day to differentiate it from the other enterococci. Partially-purified enterocin EM41 (Ent EM41, dose 40 μl/animal/day and its producer were applied to water for 21 days. The experiment lasted 35 days. Sampling was performed at days 0 - 1, 21 (3 weeks of additive application, 35 (2 weeks after cessation of additive application from every bird. E. faecium EM41 sufficiently colonized the intestinal tract of laying hens from the initial concentration 109 cfu/g, its count reached 4.30 log cfu/g at day 21. PCR genotypization confirmed the identity of the EM41 strain with the species Enterococcus faecium. E. faecium EM41 and its enterocin EM41 showed antimicrobial effects demonstrated by reduction of coagulase-positive and coagulase-negative staphylococci, coliforms, Pseudomonas spp., Aeromonas spp. and Campylobacter spp. The hens were Salmonella spp. free. After administration of both additives, phagocytic activity was stimulated with a significant increase. The additives did not negatively influence biochemical and haematological parameters or weight gains.
Al Kassaa, I.; Safourim, N.; Mostafa, N.; Hamze, M.
Lactic acid bacteria (LAB) are used in many fields such as fermentation agents, increasing nutritional value and improving organoleptic quality of food. Also they are use das probiotics and preservatives against pathogensand spoilage microbes by producing antimicrobial substances such as bacteriocins. Fifty cow's milk samples were collected and 175 LAB isolates were isolated and identified by using biochemical method. Fifteen isolates showedan antimicrobial activity against Listeria monocytogenesATCC® 19115™.One strain, BL4 which showed the strongest activity, was chosen to extract and characterize its antimicrobial substance in order to evaluate its potential use as a new food protective agent. This strain was identified as Enterococcus mundtiibypyrosequencing method.The active substance was extracted using solvent method.This Bacteriocin likeInhibitory Substances “BLIS”can support a high temperature (121 °C) for a long time and resist pH variation.The BLIS BL4 can be considered as a peptide active against many food pathogen and food-spoilage microbes, such as Listeriamonocytogenes and Penicillium spp. BLISBL4can be used in food application as bio-preservative to reduce food-spoilage and food-borne diseases in food product. (author
Determination of antimicrobial resistance of Enterococcus strains isolated from pigs and their genotypic characterization by method of amplification of DNA fragments surrounding rare restriction sites (ADSRRS fingerprinting).
Nowakiewicz, Aneta; Ziółkowska, Grażyna; Trościańczyk, Aleksandra; Zięba, Przemysław; Gnat, Sebastian
In this study, we analysed phenotypic resistance profiles and their reflection in the genomic profiles of Enterococcus spp. strains isolated from pigs raised on different farms. Samples were collected from five pig farms (n=90 animals) and tested for Enterococcus. MICs of 12 antimicrobials were determined using the broth microdilution method, and epidemiological molecular analysis of strains belonging to selected species (faecalis, faecium and hirae) was performed using the ADSRRS-fingerprinting (amplification of DNA fragments surrounding rare restriction sites) method with a few modifications. The highest percentage of strains was resistant to tetracycline (73.4 %), erythromycin and tylosin (42.5 %) and rifampin (25.2 %), and a large number of strains exhibited high-level resistance to both kanamycin (25.2 %) and streptomycin (27.6 %). The strains of E. faecalis, E. faecium and E. hirae (n=184) revealed varied phenotypic resistance profiles, among which as many as seven met the criteria for multidrug resistance (30.4 % of strains tested). ADSRRS-fingerprinting analysis produced 17 genotypic profiles of individual strains which were correlated with their phenotypic resistance profiles. Only E. hirae strains susceptible to all of the chemotherapeutics tested had two different ADSRRS profiles. Moreover, eight animals were carriers of more than one genotype belonging to the same Enterococcus spp., mainly E. faecalis. Given the possibility of transmission to humans of the high-resistance/multidrug resistance enterococci and the significant role of pigs as food animals in this process, it is necessary to introduce a multilevel control strategy by carrying out research on the resistance and molecular characteristics of indicator bacterial strains isolated from animals on individual farms.
Lidya Morita Sondang
Tujuan penelitian ini adalah untuk mengetahui tingkat ketahanan 2 klon Eucalyptus spp yaitu Eucalyptus grandis x Eucalyptus pellita dan Eucalyptus grandis x Eucalyptus urophylla terhadap Mycosphaerella spp serta mengetahui virulensi Mycospaherella spp pada 2 kelas umur (2 dan 3 bulan) pada tanaman Eucalyptus spp. Penelitian ini dilaksanakan dengan pengambilan sampel bibit tanaman Eucalyptus grandis x Eucalyptus pellita dan Eucalyptus grandis x Eucalyptus urophylla dari pembibitan PT.Toba Pulp...
Punj, Sumit; John, Gilbert H.
Azo dyes are widely used in the food, pharmaceutical, paper, and textile industries. Some azo dyes are known to produce carcinogenic compounds upon reductive cleavage of the azo bond (N=N) by intestinal flora. There is not much information available on the effect of these dyes on the physiology of the gut microflora as well as their kinetics of reduction in different environments. The azoreductase activity of Enterococcus faecalis, an important opportunistic intestinal pathogen, was tested us...
Ana Paula da Silva
Full Text Available Foram obtidas 154 amostras de lesões cutâneas de cães com pioderma superficial atendidos no Serviço de Dermatologia Veterinária do Hospital Veterinário Universitário (HVU da Universidade Federal de Santa Maria (UFSM, com o objetivo de determinar a suscetibilidade antimicrobiana de isolados de Staphylococcus spp. e avaliar a presença de multirresistência. Após isolamento e identificação, as cepas foram submetidas ao teste de sensibilidade aos antimicrobianos, cujos resultados evidenciaram menores percentuais de resistência à associação amoxicilina e ácido clavulânico (1,9%, cefadroxil (1,9%, cefalexina (1,9% e vancomicina (0,6%. Os maiores percentuais de resistência foram frente à amoxicilina (60,4% e penicilina G (60,4%. A multirresistência foi detectada em 23,4% e a resistência à meticilina em 5,8% das amostras. Pode-se concluir que os isolados de Staphylococcus spp. apresentam elevada suscetibilidade aos antimicrobianos comumente utilizados no tratamento dos piodermas superficiais em cães atendidos no HVU-UFSM, como a cefalexina e a amoxicilina associada ao ácido clavulânico, confirmando a eleição desses fármacos para o tratamento de cães com esta afecção. A suscetibilidade diminuída das cepas frente às fluoroquinolonas, também recomendadas pela literatura para o tratamento de pioderma, permite sugerir que estes fármacos não devem mais ser considerados na seleção empírica. A identificação de Staphylococcus spp. multirresistentes na população canina estudada justifica análises bacteriológicas periódicas e regionais de lesões cutâneas de cães com pioderma superficial, a fim de minimizar a seleção de bactérias resistentes, possíveis falhas terapêuticas e também motiva a antimicrobianoterapia prudente.
Mikalsen, Theresa; Pedersen, Torunn; Willems, Rob; Coque, Teresa M; Werner, Guido; Sadowy, Ewa; van Schaik, Willem; Jensen, Lars Bogø; Sundsfjord, Arnfinn; Hegstad, Kristin
The success of Enterococcus faecium and E. faecalis evolving as multi-resistant nosocomial pathogens is associated with their ability to acquire and share adaptive traits, including antimicrobial resistance genes encoded by mobile genetic elements (MGEs). Here, we investigate this mobilome in successful hospital associated genetic lineages, E. faecium sequence type (ST)17 (n=10) and ST78 (n=10), E. faecalis ST6 (n=10) and ST40 (n=10) by DNA microarray analyses. The hybridization patterns of 272 representative targets including plasmid backbones (n=85), transposable elements (n=85), resistance determinants (n=67), prophages (n=29) and clustered regularly interspaced short palindromic repeats (CRISPR)-cas sequences (n=6) separated the strains according to species, and for E. faecalis also according to STs. RCR-, Rep_3-, RepA_N- and Inc18-family plasmids were highly prevalent and with the exception of Rep_3, evenly distributed between the species. There was a considerable difference in the replicon profile, with rep 17/pRUM , rep 2/pRE25 , rep 14/EFNP1 and rep 20/pLG1 dominating in E. faecium and rep 9/pCF10 , rep 2/pRE25 and rep 7 in E. faecalis strains. We observed an overall high correlation between the presence and absence of genes coding for resistance towards antibiotics, metals, biocides and their corresponding MGEs as well as their phenotypic antimicrobial susceptibility pattern. Although most IS families were represented in both E. faecalis and E. faecium, specific IS elements within these families were distributed in only one species. The prevalence of IS256-, IS3-, ISL3-, IS200/IS605-, IS110-, IS982- and IS4-transposases was significantly higher in E. faecium than E. faecalis, and that of IS110-, IS982- and IS1182-transposases in E. faecalis ST6 compared to ST40. Notably, the transposases of IS981, ISEfm1 and IS1678 that have only been reported in few enterococcal isolates were well represented in the E. faecium strains. E. faecalis ST40 strains harboured
Full Text Available Enterococci are commensally bacteria of the gastrointestinal and female genital tract in humans and some mammals and birds, and one of the significant causes of hospital-acquired infections, especially in immuno-compromised patients. Genetic fingerprinting (DNA fingerprinting is a tool for identifying, marking and prevention of infectious agents dissemination. SSR (short sequence repeat are known to suffer frequent variations in the number of repetitive units.MLVA (multiple locus variable number tandem repeats analysis is a variant of genetic fingerprinting, in epidemiological studies on the pathogenetic Enterococcus faecium. Our study included laboratory Enterococcus faecium strains or isolated from clinical cases or from the environment (2003-2008. All analyzed strains of Enterococcus faecium were sensitive to vancomycin, except BM4147, and resistant to oxacilin. Strains isolated from the birds’ samples have shown a smaller resistance profile than those of human origin. 33 Enterococus faecium strains were analyzed by PCR amplification. 27 MT (VNTR profiles were obtained: six in the case of the strains isolated from birds, 15 in the case of the strains isolated form humans, 4 in the case of the collection strains and 2 in the case of the strains isolated from water samples. Among the strains isolated from humans and those isolated from animals, identical profiles were not recorded. Within the strains isolated from clinical cases, and those isolated from birds, circulating genotypes were noted, which can be considered as epidemical. The strains used as probiotics proved to be different from those circulating in birds. All MLVA profiles codes compared with those published on line in the UMC Utrecht database proved to be different. Results obtained in this study support the usefulness of the polymorphic VNTR analysis, as genetic marker, inepidemiological investigations.
Godfred Ngu Tanih
Full Text Available Objective: To evaluate the prevalence of Enterococcus species in cattle faeces, their corresponding drug resistant patterns as well as the genes coding for resistance in the isolates. Methods: Two hundred and ninety rectal swabs were cultured for the isolation of Enterococcus. Presumptive isolates were confirmed by PCR, targeting the tuf gene, and confirmed isolates were identified to species level, using species-specific primers aimed at targeting six different species. Additionally, antibiogram was performed by disc diffusion and genes implicated in resistance were evaluated using molecular methods. Results: All presumptive isolates were confirmed as Enterococcus and speciated as: Enterococcus hirae (82%, Enterococcus faecium (5%, Enterococcus durans (5%, Enterococcus faecalis (2% and 6% of unidentified species. Resistance to various antimicrobials ranged from 16.4% for penicillin to 69.6% for erythromycin. Among the tetracycline and erythromycin-resistant isolates, tet M (100% and erm B (29% were the only amplified genes known to mediate resistance respectively. Other detected genes included van B (25%, van C1 (21% and bla Z (11%. Conclusions: A high prevalence of multidrug resistant Enterococcus species was observed in this study, accentuating the need to improve on animal farming practices to prevent the dissemination of this microorganism to the environment.
Fallavollita, Luca; Di Gioacchino, Lorena; Balestrini, Fabrizio
Enterococci are common organisms associated with endocarditis, but infection by Enterococcus durans is very rare. To our knowledge, only 3 cases have been reported in the medical literature, and all 3 have involved native valves. Here we publish the first reported case (to our knowledge) of E. durans endocarditis in association with a bioprosthetic aortic valve. After the organism and its antibiotic susceptibility were identified, the 74-year-old male patient was treated successfully with teicoplanin and gentamicin, over a course of 6 weeks.
Full Text Available The diversity of multidrug-resistance MDR of pathogenic strains to antibiotics most widely used for treatment of human diseases in the Republics of Armenia and Nagorno Karabakh were examined. It was shown that difference of resistance of pathogens to antibiotics depends on their isolation sources. It was shown that bacteriocin containing partially purified preparations obtained from different strains of Enterococcus faecium and durans species isolated from various samples of matsun salted cheese and other acid milk products from milk of different domestic animals from rural households inhibited the growth of multidrug-resistant bacteria belonging to different taxonomic groups with different efficiency.
Angelidis, Apostolos S; Kalamaki, Mary S; Georgiadou, Sofia S
Agar Listeria according to Ottaviani and Agosti (ALOA) is the mandatory medium used for the detection and enumeration of Listeria monocytogenes in foods according to the official International Organization for Standardization (ISO) methods. On ALOA, Listeria spp. appear as bluish-green colonies due to the production of β-D-glucosidase, an enzyme that cleaves 5-bromo-4-chloro-3-indolyl-β-D-glucopyranoside, a chromogenic substrate included in the formulation of the medium. The present work reports on bacterial isolates (n=64) from ready-to-eat soft cheeses, which are able to grow on ALOA, forming bluish-green colonies and therefore phenotypically resemble Listeria spp. All isolates were also capable of growing on the selective media PALCAM and RAPID L'mono. The isolates were characterised with biochemical tests including those specified in the ISO standards for the confirmation of Listeria spp. and identified via partial sequencing of their 16S rRNA gene. According to sequencing results the isolates represented 12 different bacterial species or species-groups belonging to seven different genera: Bacillus spp. (B. circulans, B. clausii, B. licheniformis and B. oleronius), Cellulosimicrobium spp. (C. funkei), Enterococcus spp. (E. faecalis, E. faecium/durans), Kocuria spp. (K. kristinae), Marinilactibacillus spp. (M. psychrotolerans), Rothia spp. (R. terrae) and Staphylococcus spp. (S. sciuri and S. saprophyticus subsp. saprophyticus/xylosus). Cellulosimicrobium spp. have never been previously isolated from foods. These results significantly extend the list of bacteria previously known as capable of growing on ALOA as bluish-green colonies and suggest that there may be room for further improvement in the medium's inhibitory properties towards non-Listeria spp., Gram-positive bacteria present in foods. Copyright © 2014 Elsevier B.V. All rights reserved.
Boehnke, B; Karlovsky, P; Pfohl, K; Gamliel, A; Isack, Y; Dehne, H W
In 2013 Allium cepa bulbs from different fields in Northern and Southern Germany, seeds and sets from onion breeders were analysed for infestation with Fusarium species. The same investigation was done in 2014 with different edible Allium spp. from local markets. Different Fusarium spp. were isolated and identified by morphological characterisation. 24 different Fusarium spp. were identified. The diversity of Fusarium spp. and the intensity of infestation was higher on edible bulbs compared to the younger sets and seeds. The analysed onions and other edible Allium spp. from local markets showed also high contents of different Fusarium species. The most prevalent identified Fusarium sp. in the analysed Allium spp. in Germany was Fusarium oxysporum which can cause the Fusarium Basal Rot, followed by Fusarium solani. Fusarium proliferatum, which can cause the Fusarium Salmon Blotch in onions, could be detected in about half of the sampled onion fields and in approximately 10% of all analysed onions from fields. Also in the onion sets, on the surface of the seeds and in other edible Allium spp. F. proliferatum could be identified. Besides F. proliferatum, further mycotoxin producing Fusarium spp. like Fusarium equiseti or Fusarium tricinctum were identified. Other Fusarium spp. like Fusarium sporotrichioides and Fusarium poae were first described in Allium sp. in this study. The two most prevalent Fusarium spp. F. oxysporum and F. solani are able to produce mycotoxins like enniatins, fumonisins, moniliformin and T-2 toxins. Fusarium sp. like F. proliferatum, F. equiseti and F. tricinctum are able to produce additional toxins like beauvericins, zearalenone and diacetoscirpenol. This high number of Fusarium spp., which are able to produce a broad spectrum of different mycotoxins, could be a potential health risk for human beings and livestock.
Full Text Available Irrigation with wastewater releases pharmaceuticals, pathogenic bacteria, and resistance genes, but little is known about the accumulation of these contaminants in the environment when wastewater is applied for decades. We sampled a chronosequence of soils that were variously irrigated with wastewater from zero up to 100 years in the Mezquital Valley, Mexico, and investigated the accumulation of ciprofloxacin, enrofloxacin, sulfamethoxazole, trimethoprim, clarithromycin, carbamazepine, bezafibrate, naproxen, diclofenac, as well as the occurrence of Enterococcus spp., and sul and qnr resistance genes. Total concentrations of ciprofloxacin, sulfamethoxazole, and carbamazepine increased with irrigation duration reaching 95% of their upper limit of 1.4 µg/kg (ciprofloxacin, 4.3 µg/kg (sulfamethoxazole, and 5.4 µg/kg (carbamazepine in soils irrigated for 19-28 years. Accumulation was soil-type-specific, with largest accumulation rates in Leptosols and no time-trend in Vertisols. Acidic pharmaceuticals (diclofenac, naproxen, bezafibrate were not retained and thus did not accumulate in soils. We did not detect qnrA genes, but qnrS and qnrB genes were found in two of the irrigated soils. Relative concentrations of sul1 genes in irrigated soils were two orders of magnitude larger (3.15 × 10(-3 ± 0.22 × 10(-3 copies/16S rDNA than in non-irrigated soils (4.35 × 10(-5± 1.00 × 10(-5 copies/16S rDNA, while those of sul2 exceeded the ones in non-irrigated soils still by a factor of 22 (6.61 × 10(-4 ± 0.59 × 10(-4 versus 2.99 × 10(-5 ± 0.26 × 10(-5 copies/16S rDNA. Absolute numbers of sul genes continued to increase with prolonging irrigation together with Enterococcus spp. 23S rDNA and total 16S rDNA contents. Increasing total concentrations of antibiotics in soil are not accompanied by increasing relative abundances of resistance genes. Nevertheless, wastewater irrigation enlarges the absolute concentration of resistance genes in soils due to a
Siebe, Christina; Willaschek, Elisha; Sakinc, Tuerkan; Huebner, Johannes; Amelung, Wulf; Grohmann, Elisabeth; Siemens, Jan
Irrigation with wastewater releases pharmaceuticals, pathogenic bacteria, and resistance genes, but little is known about the accumulation of these contaminants in the environment when wastewater is applied for decades. We sampled a chronosequence of soils that were variously irrigated with wastewater from zero up to 100 years in the Mezquital Valley, Mexico, and investigated the accumulation of ciprofloxacin, enrofloxacin, sulfamethoxazole, trimethoprim, clarithromycin, carbamazepine, bezafibrate, naproxen, diclofenac, as well as the occurrence of Enterococcus spp., and sul and qnr resistance genes. Total concentrations of ciprofloxacin, sulfamethoxazole, and carbamazepine increased with irrigation duration reaching 95% of their upper limit of 1.4 µg/kg (ciprofloxacin), 4.3 µg/kg (sulfamethoxazole), and 5.4 µg/kg (carbamazepine) in soils irrigated for 19–28 years. Accumulation was soil-type-specific, with largest accumulation rates in Leptosols and no time-trend in Vertisols. Acidic pharmaceuticals (diclofenac, naproxen, bezafibrate) were not retained and thus did not accumulate in soils. We did not detect qnrA genes, but qnrS and qnrB genes were found in two of the irrigated soils. Relative concentrations of sul1 genes in irrigated soils were two orders of magnitude larger (3.15×10−3±0.22×10−3 copies/16S rDNA) than in non-irrigated soils (4.35×10−5±1.00×10−5 copies/16S rDNA), while those of sul2 exceeded the ones in non-irrigated soils still by a factor of 22 (6.61×10–4±0.59×10−4 versus 2.99×10−5±0.26×10−5 copies/16S rDNA). Absolute numbers of sul genes continued to increase with prolonging irrigation together with Enterococcus spp. 23S rDNA and total 16S rDNA contents. Increasing total concentrations of antibiotics in soil are not accompanied by increasing relative abundances of resistance genes. Nevertheless, wastewater irrigation enlarges the absolute concentration of resistance genes in soils due to a long-term increase in
Impacto de la resistencia a la meticilina sobre la mortalidad y vigilancia de la sensibilidad a la vancomicina en bacteriemias causadas por Staphylococcus aureus Impact of methicillin resistance on mortality and surveillance of vancomycin susceptibility in bacteremias caused by Staphylococcus aureus
Full Text Available Staphylococcus aureus es uno de los principales patógenos nosocomiales y produce una alta morbimortalidad en numerosos hospitales del mundo. Además, la incidencia de bacteriemias por este microorganismo ha aumentado significativamente en las últimas décadas. Los objetivos del presente trabajo fueron identificar los factores de riesgo que favorecen la aparición de resistencia a la meticilina en aislamientos de S. aureus y los factores que afectan la mortalidad por bacteriemias asociadas a este patógeno, así como evaluar la sensibilidad a la vancomicina de las cepas resistentes a la meticilina. Se estudiaron 39 aislamientos de S. aureus provenientes de hemocultivos de pacientes internados con bacteriemia en la Nueva Clínica Chacabuco de Tandil (Pcia. de Buenos Aires, Argentina en el período 01/2006-12/2008. La mortalidad global fue del 51,3% y estuvo significativamente asociada con la resistencia a la meticilina (OR: 4,20; IC95%: 1,08-16,32; p: 0,05; aunque dicho factor no fue un predictor independiente de mortalidad. La cirugía previa (OR: 17,23; IC95%: 1,80-164,60 y la estancia previa en la unidad de cuidados intensivos (OR: 21,12; IC95%: 2,33-191,30 fueron predictores independientes de la resistencia a la meticilina y la asistencia respiratoria mecánica (OR: 15,99; IC: 3,24-78,86 fue un predictor independiente de la mortalidad. No se detectaron cepas con sensibilidad disminuida a la vancomicina. Todos los aislamientos estudiados fueron sensibles in vitro a la vancomicina, con una CIM50 y una CIM90 de 0,5 μg/ml.Staphylococcus aureus is a major nosocomial pathogen that causes severe morbidity and mortality in many hospitals worldwide. Besides, the incidence of S. aureus bacteremia has significantly increased over the past decades. The aims of this study were to detect the risk factors for methicillin resistance and mortality and to evaluate vancomycin susceptibility in methicillin-resistant isolates. Thus, 39 S. aureus isolates from
Amaral, Daniel M F; Silva, Luana F; Casarotti, Sabrina N; Nascimento, Liane Caroline Sousa; Penna, Ana Lúcia B
In this study, we evaluated the survival of Enterococcus faecium and Enterococcus durans, isolated from cheese, in the presence of medications and under simulated in vitro gastrointestinal conditions. The presence of genes encoding virulence factors, the susceptibility to antimicrobial agents, and adhesion properties were also assessed. Enterococcus faecium and E. durans both exhibited resistance to most of the tested medications but showed a large sensitivity to analgesics and antihypertensives; they also showed wide susceptibility to antimicrobial agents. Enterococcus durans SJRP29 had greater resistance to the presence of medications in comparison with the probiotic Lactobacillus acidophilus La-5. The strains, except for E. durans SJRP05, did not harbor virulence genes. Enterococcus durans SJRP14, SJRP17, and SJRP26 were sensitive to all tested antimicrobial agents. Enterococcus faecium was more stable during the simulation of gastrointestinal tract and showed greater viability. At the end of the assay, except for E. durans SJRP17, all strains showed high viability (>7 log cfu/mL). Enterococcus durans SJRP29 stood out from the other strains and was selected for further evaluation; it tolerated up to 3.0% NaCl at 30 and 37°C, besides having good adhesion properties (high values of auto-aggregation, co-aggregation, and hydrophobicity). Additionally, the microorganism did not show bile salt hydrolase activity or mucin degradation. These results encourage carrying out additional tests to evaluate the probiotic features by using in vitro dynamic models and in vivo tests before applying these strains to a food system. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.
Kamelia Mahmoud Osman
that aquatic environments are correlated to proximity to anthropogenic activities in determining the antimicrobial resistance patterns of Enterococcus spp. recovered from fish in the river Nile in Giza, Elmounib, Egypt as a continuation of our larger study on the reservoirs of antibiotic resistance in the environment.
Full Text Available Normal 0 21 false false false SK JA X-NONE Biogenic amines (BA are basic nitrogenous compounds formed mainly by decarboxylation of amino acids. There are generated in course of microbial, vegetable and animal metabolisms. The aim of the study was to monitor factors affected production of biogenic amines by Enterococcus faecium, which is found in rabbit meat. Biogenic amines were analyzed by means of UPLC (ultrahigh performance liquid chromatography equipped with a UV/VIS DAD detector. Decarboxylation activity of E. faecium was mainly influenced by the cultivation temperature and the amount of NaCl in this study. E. faecium produced most of the monitored biogenic amines levels: tyramine ˂2500 mg.l-1; putrescine ˂30 mg.l-1; spermidine ˂10 mg.l-1 and cadaverine ˂5 mg.l-1.doi:10.5219/182
Madsen, Kristian T; Skov, Marianne N; Gill, Sabine
INTRODUCTION: The enterococci are accountable for up to 20% of all cases of infective endocarditis, with Enterococcus faecalis being the primary causative isolate. Infective endocarditis is a life-threatening infection of the endocardium that results in the formation of vegetations. Based...... on a literature review, this paper provides an overview of the virulence factors associated with E. faecalis infective endocarditis. Furthermore, it reports the effects of active or passive immunization against some of these involved factors. INDIVIDUAL VIRULENCE FACTORS: Nine virulence factors have in particular...... been associated with E. faecalis infective endocarditis. Absence of these factors entailed attenuation of strains in both mixed- and mono-bacterial infection endocarditis models as well as in in vitro and ex vivo assays when compared to their virulence factor expressing parental strains. PATHOGENESIS...
Bortolini Maria Cecilia
Full Text Available Aim : Evaluate in vitro the intratubular penetration and permeability of endodontic sealers in teeth contaminated with Enterococcus faecalis. Materials and Methods : Human canines were filled with AHPlus ® , Endo CPM-sealer ® or EndoRez ® sealers. To evaluate permeability, the coronary portion of each tooth was contaminated with E. faecalis, then the apical portion was immersed in brain heart infusion (BHI broth, and medium turbidity was observed for thirty days. Scanning electron microscope (SEM was used to evaluate the intratubular penetration of each sealer at the cervical, middle, and apical thirds of the tooth. Results : Only one tooth from the Endo CPM-sealer ® group presented broth contamination. EndoRez ® showed increased intratubular penetration compared to AHPlus ® and Endo CPM-sealer ® . Conclusions : Endo CPM-sealer ® showed greater permeability to E. faecalis and EndoRez ® showed increased intratubular penetration.
Hullahalli, Karthik; Rodrigues, Marinelle; Palmer, Kelli L
CRISPR-Cas provides a barrier to horizontal gene transfer in prokaryotes. It was previously observed that functional CRISPR-Cas systems are absent from multidrug-resistant (MDR) Enterococcus faecalis , which only possess an orphan CRISPR locus, termed CRISPR2, lacking cas genes. Here, we investigate how the interplay between CRISPR-Cas genome defense and antibiotic selection for mobile genetic elements shapes in vitro E. faecalis populations. We demonstrate that CRISPR2 can be reactivated for genome defense in MDR strains. Interestingly, we observe that E. faecalis transiently maintains CRISPR targets despite active CRISPR-Cas systems. Subsequently, if selection for the CRISPR target is present, toxic CRISPR spacers are lost over time, while in the absence of selection, CRISPR targets are lost over time. We find that forced maintenance of CRISPR targets induces a fitness cost that can be exploited to alter heterogeneous E. faecalis populations.
Lester, Camilla H; Sandvang, Dorthe; Olsen, Stefan
BACKGROUND: Ampicillin-resistant Enterococcus faecium isolates are reported in increasing numbers in many European hospitals. The clonal complex 17 (CC17) characterized by ampicillin resistance has been associated with nosocomial E. faecium outbreaks and infections in five continents. The aim...... in the number of infections caused by enterococci was observed from 2002 through 2006. The increase was mainly caused by E. faecium isolates, which tripled, whereas the number of E. faecalis isolates increased by only 23% during the same period. There was also a significant increase in the number of ampicillin......-resistant E. faecium isolates. MLST showed that 98% of the tested ampicillin-resistant E. faecium isolates belonged to CC17. PFGE showed eight different clusters and we found indications of clonal spread within the hospitals. CONCLUSIONS: Ampicillin-resistant E. faecium isolates have increased in frequency...
Liu, Hui; Zhang, Lanwei; Yi, Huaxi; Han, Xue; Gao, Wei; Chi, Chunliang; Song, Wei; Li, Haiying; Liu, Chunguang
An enterocin-producing Enterococcus faecium T1 was isolated from Chinese Tibet cheese. The enterocin was purified by SP-Sepharose and reversed phase HPLC. It was identified as unique from other reported bacteriocins based on molecular weight (4629 Da) and amino acid compositions; therefore it was subsequently named enterocin T1. Enterocin T1 was stable at 80-100 °C and over a wide pH range, pH 3.0-10.0. Protease sensitivity was observed to trypsin, pepsin, papain, proteinase K, and pronase E. Importantly, enterocin T1 was observed to inhibit the growth of numerous Gram-negative and Gram-positive bacteria including Pseudomonas putida, Pseudomonas aeruginosa, Pseudomonas fluorescens, Escherichia coli, Salmonella typhimurium, Shigella flexneri, Shigella sonnei, Staphylococcus aureus, Listeria monocytogenes. Take together, these results suggest that enterocin T1 is a novel bacteriocin with the potential to be used as a bio-preservative to control Pseudomonas spp. in food.
Paganelli, Fernanda L.; van de Kamer, Tim; Brouwer, Ellen C.; Leavis, Helen L.; Woodford, Neil; Bonten, Marc J M; Willems, Rob J L; Hendrickx, Antoni P A
Enterococcus faecium is a multidrug-resistant (MDR) nosocomial pathogen causing significant morbidity in debilitated patients. New antimicrobials are needed to treat antibiotic-resistant E. faecium infections in hospitalised patients. E. faecium incorporates lipoteichoic acid (LTA)
Castillo-Rojas, Gonzalo; Mazari-Hiríart, Marisa; Ponce de León, Sergio; Amieva-Fernández, Rosa I; Agis-Juárez, Raúl A; Huebner, Johannes; López-Vidal, Yolanda
Enterococci are part of the normal intestinal flora in a large number of mammals, and these microbes are currently used as indicators of fecal contamination in water and food for human consumption. These organisms are considered one of the primary causes of nosocomial and environmental infections due to their ability to survive in the environment and to their intrinsic resistance to antimicrobials. The aims of this study were to determine the biochemical patterns and antimicrobial susceptibilities of Enterococcus faecalis and E. faecium isolates from clinical samples and from water (groundwater, water from the Xochimilco wetland, and treated water from the Mexico City Metropolitan Area) and to determine the genetic relationships among these isolates. A total of 121 enterococcus strains were studied; 31 and 90 strains were isolated from clinical samples and water (groundwater, water from the Xochimilco wetland, and water for agricultural irrigation), respectively. Identification to the species level was performed using a multiplex PCR assay, and antimicrobial profiles were obtained using a commercial kit. Twenty-eight strains were analyzed by pulsed-field gel electrophoresis (PFGE). E. faecium strains isolated from water showed an atypical biochemical pattern. The clinical isolates showed higher resistance to antibiotics than those from water. Both the enterococci isolated from humans, and those isolated from water showed high genetic diversity according to the PFGE analysis, although some strains seemed to be closely related. In conclusion, enterococci isolated from humans and water are genetically different. However, water represents a potential route of transmission to the community and a source of antimicrobial resistance genes that may be readily transmitted to other, different bacterial species.
Full Text Available Enterococci are part of the normal intestinal flora in a large number of mammals, and these microbes are currently used as indicators of fecal contamination in water and food for human consumption. These organisms are considered one of the primary causes of nosocomial and environmental infections due to their ability to survive in the environment and to their intrinsic resistance to antimicrobials. The aims of this study were to determine the biochemical patterns and antimicrobial susceptibilities of Enterococcus faecalis and E. faecium isolates from clinical samples and from water (groundwater, water from the Xochimilco wetland, and treated water from the Mexico City Metropolitan Area and to determine the genetic relationships among these isolates. A total of 121 enterococcus strains were studied; 31 and 90 strains were isolated from clinical samples and water (groundwater, water from the Xochimilco wetland, and water for agricultural irrigation, respectively. Identification to the species level was performed using a multiplex PCR assay, and antimicrobial profiles were obtained using a commercial kit. Twenty-eight strains were analyzed by pulsed-field gel electrophoresis (PFGE. E. faecium strains isolated from water showed an atypical biochemical pattern. The clinical isolates showed higher resistance to antibiotics than those from water. Both the enterococci isolated from humans, and those isolated from water showed high genetic diversity according to the PFGE analysis, although some strains seemed to be closely related. In conclusion, enterococci isolated from humans and water are genetically different. However, water represents a potential route of transmission to the community and a source of antimicrobial resistance genes that may be readily transmitted to other, different bacterial species.
Joosten, H M; Nunez, M; Devreese, B; Van Beeumen, J; Marugg, J D
A simple two-step procedure was developed to obtain pure enterocin 4, a bacteriocin produced by Enterococcus faecalis INIA 4. Chemical and genetic characterization revealed that the primary structure of enterocin 4 is identical to that of peptide antibiotic AS-48 from Enterococcus faecalis S-48. In contrast to the reported inhibitory spectrum of AS-48, enterocin 4 displayed no activity against gram-negative bacteria. PMID:8900014
Joosten, H M; Nunez, M; Devreese, B; Van Beeumen, J; Marugg, J D
A simple two-step procedure was developed to obtain pure enterocin 4, a bacteriocin produced by Enterococcus faecalis INIA 4. Chemical and genetic characterization revealed that the primary structure of enterocin 4 is identical to that of peptide antibiotic AS-48 from Enterococcus faecalis S-48. In contrast to the reported inhibitory spectrum of AS-48, enterocin 4 displayed no activity against gram-negative bacteria.
Dolka, Beata; Boyen, Filip; Butaye, Patrick
Here, we report the draft genome sequences of two commensal Enterococcus cecorum strains (1710s23 and 1711s24), cultivated from the ceca of healthy laying hens originating from different farms in Belgium.......Here, we report the draft genome sequences of two commensal Enterococcus cecorum strains (1710s23 and 1711s24), cultivated from the ceca of healthy laying hens originating from different farms in Belgium....
Rathe, Mathias; Kristensen, Lise; Ellermann-Eriksen, Svend
Vancomycin-resistant enterococci (VRE) have emerged to become a significant nosocomial pathogen. However, detection may be challenging and treatment possibilities are limited. Reports of resistance to linezolide, daptomycin and tigecycline underline the need for reliable susceptibility testing wi...
Dwiastuti, Mutia Erti; Fajri, Melisa N; Yunimar, Yunimar
Layu yang disebabkan oleh Fusarium spp. merupakan salah satu penyakit penting tanaman stroberi (Fragaria x ananassa Dutch.) di daerah subtropika, yang dapat menggagalkan panen. Penelitian bertujuan untuk mempelajari potensi Trichoderma spp. dalam mengendalikan Fusarium spp. Isolat Trichoderma spp. diisolasi dari rizosfer tanaman stroberi dan Fusarium spp. diisolasi dari tanaman stroberi yang mengalami layu fusarium. Isolat cendawan dimurnikan, dikarakterisasi, dan dibandingkan dengan isolat c...
Full Text Available Objective: to investigate the prevalence and antimicrobial resistance of Enterococcus species isolated from a university hospital, and explore the mechanisms underlying the antimicrobial resistance, so as to provide clinical evidence for the inappropriate clinical use of antimicrobial agents and the control and prevention of enterococcal infections. Methods: a total of 1,157 enterococcal strains isolated from various clinical specimens from January 2010 to December 2012 in the General Hospital of Ningxia Medical University were identified to species level with a VITEK-2 COMPACT fully automated microbiological system, and the antimicrobial susceptibility of Enterococcus species was determined using the Kirby-Bauer disc diffusion method. The multiple-drug resistant enterococcal isolates were screened from the clinical isolates of Enterococcus species from the burns department. The minimal inhibitory concentration (MIC of Enterococcus species to the three fluoroquinolones, including ciprofloxacin, gatifloxacin and levofloxacin was determined with the agar dilution method, and the changes in the MIC of Enterococcus species to the three fluoroquinolones following reserpine treatment were evaluated. The β-lactam, aminoglycoside, tetracycline, macrolide, glycopeptide resistance genes and the efflux pump emeA genes were detected in the enterococcal isolates using a polymerase chain reaction (PCR assay. Results: the 1,157 clinical isolates of Enterococcus species included 679 E. faecium isolates (58.7%, 382 E. faecalis isolates (33%, 26 E. casseliflavus isolates (2.2%, 24 E. avium isolates (2.1%, and 46 isolates of other Enterococcus species (4%. The prevalence of antimicrobial resistance varied significantly between E. faecium and E. faecalis, and ≤1.1% of these two Enterococcus species were found to be resistant to vancomycin, teicoplanin or linezolid. In addition, the Enterococcus species isolated from different departments of the hospital
Full Text Available With growing concern over bacterial resistance, the identification of new antimicrobial means is paramount. In the oral cavity microorganisms are essential to the development of periradicular diseases and are the major causative factors associated with endodontic treatment failure. As quaternary ammonium compounds have the ability to kill a wide array of bacteria through electrostatic interactions with multiple anionic targets on the bacterial surface, it is likely that they can overcome bacterial resistance. Melding these ideas, we investigated the potency of a novel endodontic sealer in limiting Enterococcus faecalis growth. We used a polyethyleneimine scaffold to synthesize nano-sized particles, optimized for incorporation into an epoxy-based endodontic sealer. The novel endodontic sealer was tested for its antimicrobial efficacy and evaluated for biocompatibility and physical eligibility. Our results show that the novel sealer foundation affixes the nanoparticles, achieving surface bactericidal properties, but at the same time impeding nanoparticle penetration into eukaryotic cells and thereby mitigating a possible toxic effect. Moreover, adequate physical properties are maintained. The nanosized quaternary amine particles interact within minutes with bacteria, triggering cell death across wide pH values. Throughout this study we demonstrate a new antibacterial perspective for endodontic sealers; a novel antibacterial, effective and safe antimicrobial means.
Full Text Available Analysis of the culture supernatant exoproteins produced by two PFGE clusters of high-level gentamicin and ciprofloxacin-resistant clinical isolates of Enterococcus faecalis from the UK and Ireland revealed two distinct protein profiles. This grouping distinguished OG1RF and GelE metalloprotease-expressing isolates from JH2-2 and other GelE-negative isolates. The integrity of the fsrABDC operon was found to determine the exoproteome composition, since an fsrB mutant of strain OG1RF appeared very similar to that of strain JH2-2, and complementation of the latter with the fsrABDC operon produced an OG1RF-like exoproteome. The proteins present in the supernatant fraction of OG1RF were separated using 2D gels and identified by mass spectrometry and comprised many mass and pI variants of the GelE and SprE proteases. In addition cell wall synthesis and cell division proteins were identified. An OG1RF fsrB mutant had a distinct exoprotein fraction with an absence of the Fsr-regulated proteases and was characterised by general stress and glycolytic proteins. The exoproteome of the OG1RF fsrB mutant resembles that of a divIVA mutant of E. faecalis, suggestive of a stress phenotype.
Cortes-Perez, Naima G; Dumoulin, Romain; Gaubert, Stéphane; Lacoux, Caroline; Bugli, Francesca; Martin, Rebeca; Chat, Sophie; Piquand, Kevin; Meylheuc, Thierry; Langella, Philippe; Sanguinetti, Maurizio; Posteraro, Brunella; Rigottier-Gois, Lionel; Serror, Pascale
Mechanisms underlying the transition from commensalism to virulence in Enterococcus faecalis are not fully understood. We previously identified the enterococcal leucine-rich protein A (ElrA) as a virulence factor of E. faecalis. The elrA gene is part of an operon that comprises four other ORFs encoding putative surface proteins of unknown function. In this work, we compared the susceptibility to phagocytosis of three E. faecalis strains, including a wild-type (WT), a ΔelrA strain, and a strain overexpressing the whole elr operon in order to understand the role of this operon in E. faecalis virulence. While both WT and ΔelrA strains were efficiently phagocytized by RAW 264.7 mouse macrophages, the elr operon-overexpressing strain showed a decreased capability to be internalized by the phagocytic cells. Consistently, the strain overexpressing elr operon was less adherent to macrophages than the WT strain, suggesting that overexpression of the elr operon could confer E. faecalis with additional anti-adhesion properties. In addition, increased virulence of the elr operon-overexpressing strain was shown in a mouse peritonitis model. Altogether, our results indicate that overexpression of the elr operon facilitates the E. faecalis escape from host immune defenses.
Pileggi, Giorgio; Wataha, John C; Girard, Myriam; Grad, Iwona; Schrenzel, Jacques; Lange, Norbert; Bouillaguet, Serge
In dentistry, residual infection remains a major cause of failure after endodontic treatment; many of these infections involve Enterococcus faecalis. In the current study, we explored the possibility that blue light activated photosensitizers could be used, in principle, to inactivate this microbe as an adjunct disinfection strategy for endodontic therapy. Three blue light absorbing photosensitizers, eosin-Y, rose bengal, and curcumin, were tested on E. faecalis grown in planktonic suspensions or biofilms. Photosensitizers were incubated for 30 min with bacteria then exposed to blue light (450-500 nm) for 240 s. Sodium hypochlorite (3%) was used as a control. After 48 h, the viability of E. faecalis was estimated by measuring colony-forming units post-exposure vs. untreated controls (CFU/mL). Blue light irradiation alone did not alter E. faecalis viability. For planktonic cultures, blue light activated eosin-Y (5 μM), rose bengal (1 μM), or curcumin (5 μM) significantly (pcurcumin of 100, 10, and 10 μM respectively, completely suppressed E. faecalis viability (p<0.05). Although the current results are limited to an in vitro model, they support further exploration of blue light activated antimicrobials as an adjunct therapy in endodontic treatment. Copyright © 2012 Elsevier B.V. All rights reserved.
Oyama, Linda B.; Crochet, Jean-Adrien; Edwards, Joan E.; Girdwood, Susan E.; Cookson, Alan R.; Fernandez-Fuentes, Narcis; Hilpert, Kai; Golyshin, Peter N.; Golyshina, Olga V.; Privé, Florence; Hess, Matthias; Mantovani, Hilario C.; Creevey, Christopher J.; Huws, Sharon A.
Antimicrobial peptides (AMPs) are gaining popularity as alternatives for treatment of bacterial infections and recent advances in omics technologies provide new platforms for AMP discovery. We sought to determine the antibacterial activity of a novel antimicrobial peptide, buwchitin, against Enterococcus faecalis. Buwchitin was identified from a rumen bacterial metagenome library, cloned, expressed and purified. The antimicrobial activity of the recombinant peptide was assessed using a broth microdilution susceptibility assay to determine the peptide's killing kinetics against selected bacterial strains. The killing mechanism of buwchitin was investigated further by monitoring its ability to cause membrane depolarization (diSC3(5) method) and morphological changes in E. faecalis cells. Transmission electron micrographs of buwchitin treated E. faecalis cells showed intact outer membranes with blebbing, but no major damaging effects and cell morphology changes. Buwchitin had negligible cytotoxicity against defibrinated sheep erythrocytes. Although no significant membrane leakage and depolarization was observed, buwchitin at minimum inhibitory concentration (MIC) was bacteriostatic against E. faecalis cells and inhibited growth in vitro by 70% when compared to untreated cells. These findings suggest that buwchitin, a rumen derived peptide, has potential for antimicrobial activity against E. faecalis.
Linda B. Oyama
Full Text Available Antimicrobial peptides (AMPs are gaining popularity as alternatives for treatment of bacterial infections and recent advances in omics technologies provide new platforms for AMP discovery. We sought to determine the antibacterial activity of a novel antimicrobial peptide, buwchitin, against Enterococcus faecalis. Buwchitin was identified from a rumen bacterial metagenome library, cloned, expressed and purified. The antimicrobial activity of the recombinant peptide was assessed using a broth microdilution susceptibility assay to determine the peptide's killing kinetics against selected bacterial strains. The killing mechanism of buwchitin was investigated further by monitoring its ability to cause membrane depolarization (diSC3(5 method and morphological changes in E. faecalis cells. Transmission electron micrographs of buwchitin treated E. faecalis cells showed intact outer membranes with blebbing, but no major damaging effects and cell morphology changes. Buwchitin had negligible cytotoxicity against defibrinated sheep erythrocytes. Although no significant membrane leakage and depolarization was observed, buwchitin at minimum inhibitory concentration (MIC was bacteriostatic against E. faecalis cells and inhibited growth in vitro by 70% when compared to untreated cells. These findings suggest that buwchitin, a rumen derived peptide, has potential for antimicrobial activity against E. faecalis.
Full Text Available Multisystemic infections with a morphologically unusual bacterium were first observed in captive critically endangered Lister's geckos (Lepidodactylus listeri on Christmas Island in October 2014. Since then the infection was identified in another captive critically endangered lizard species, the blue-tailed skink (Cryptoblepharus egeriae and two species of invasive geckos; the four clawed gecko (Gehyra mutilata and Asian house gecko (Hemidactylus frenatus, in a wide geographic range across the east side of the island. The Gram and periodic acid-Schiff positive cocci to diplococci have a propensity to form chains surrounded by a matrix, which ultrastructurally appears to be formed by fibrillar capsular projections. The bacterium was associated with severe and extensive replacement of tissues, but minimal host inflammatory response. Attempts to grow the organism in culture and in embryonated eggs were unsuccessful. Molecular characterisation of the organism placed it as a novel member of the genus Enterococcus. Disease Risk Analyses including this organism should now be factored into conservation management actions and island biosecurity.
Baureder, Michael; Hederstedt, Lars
Little in general is known about how heme proteins are assembled from their constituents in cells. The Gram-positive bacterium Enterococcus faecalis cannot synthesize heme and does not depend on it for growth. However, when supplied with heme in the growth medium the cells can synthesize two heme proteins; catalase (KatA) and cytochrome bd (CydAB). To identify novel factors important for catalase biogenesis libraries of E. faecalis gene insertion mutants were generated using two different types of transposons. The libraries of mutants were screened for clones deficient in catalase activity using a colony zymogram staining procedure. Analysis of obtained clones identified, in addition to katA (encoding the catalase enzyme protein), nine genes distributed over five different chromosomal loci. No factors with a dedicated essential role in catalase biogenesis or heme trafficking were revealed, but the results indicate the RNA degradosome (srmB, rnjA), an ABC-type oligopeptide transporter (oppBC), a two-component signal transducer (etaR), and NADH peroxidase (npr) as being important for expression of catalase activity in E. faecalis. It is demonstrated that catalase biogenesis in E. faecalis is independent of the CydABCD proteins and that a conserved proline residue in the N-terminal region of KatA is important for catalase assembly. PMID:22590595
Full Text Available Antibiotic resistance is an ever-growing problem faced by all major sectors of health care, including dentistry. Recurrent infections related to multidrug-resistant bacteria such as methicillin-resistant Staphylococcus aureus, carbapenem-resistant Enterobacteriaceae, and vancomycin-resistant enterococci (VRE in hospitals are untreatable and question the effectiveness of notable drugs. Two major reasons for these recurrent infections are acquired antibiotic resistance genes and biofilm formation. None of the traditionally known effective techniques have been able to efficiently resolve these issues. Hence, development of a highly effective antibacterial practice has become inevitable. One example of a hard-to-eradicate pathogen in dentistry is Enterococcus faecalis, which is one of the most common threats observed in recurrent root canal treatment failures, of which the most problematic to treat are its biofilm-forming VRE strains. An effective response against such infections could be the use of bacteriophages (phages. Phage therapy was found to be highly effective against biofilm and multidrug-resistant bacteria and has other advantages like ease of isolation and possibilities for genetic manipulations. The potential of phage therapy in dentistry, in particular against E. faecalis biofilms in root canals, is almost unexplored. Here we review the efforts to develop phage therapy against biofilms. We also focus on the phages isolated against E. faecalis and discuss the possibility of using phages against E. faecalis biofilm in root canals.
Vasilchenko, A S; Rogozhin, E A; Valyshev, A V
Isolate bacteriocins from Enterococcus faecium metabolites and characterize their effect on cells of Gram positive (Listeria monocytogenes) and Gram negative (Escherichia coli) bacteria. Methods of solid-phase extraction, ion-exchange and reversed phase chromatography were applied for isolation of bacteriocins from cultural medium of bacteria MALDI time-of-flight mass-spectrometry was used for characterization of the obtained preparations. The mechanism of biological effect of peptides was evaluated using DNA-tropic dyes (SYTO 9 and PI) with subsequent registration of fluorescence spectra: Atomic-force microscopy (AFM) was used for characterization of morpho-functional reaction of target cells. Peptide fractions with mass of 1.0 - 3.0 kDa were isolated from enterococci metabolites, that inhibit the growth of indicator microorganisms. E. faecium strain exoproducts were shown to increase membrane permeability during interaction with L. monocytogenes, that results in subsequent detectable disturbance of normal cell morphology of listeria. Alterations of E. coli surface during the effect of purified peptide fraction was detected using AFM. The studies carried out have revealed the effect of bacteriocins of enterococci on microorganisms with various types of cell wall composition and have confirmed the importance of bacterial barrier structure permeability disturbance in the mechanism of antimicrobial effect of enterocins.
Full Text Available Objective: Non-steroidal anti-inflammatory drugs (NSAIDs have shown antibacterial activity in some recent studies. The aim of this study was to evaluate the antibacterial effect of diclofenac against Enterococcus faecalis (E. faecalis as a resistant endodontic bacterium in comparison with ibuprofen, calcium hydroxide and amoxicillin.Materials and Methods: The antibacterial activity of materials was evaluated using agar diffusion test and tube dilution method. Mixtures of 400 mg/ml of materials were prepared. The bacteria were seeded on 10 Muller-Hinton agar culture plates. Thirty microliter of each test material was placed in each well punched in agar plates. After incubation, the zone of bacterial inhibition was measured. Minimum inhibitory concentration (MIC of the test materials was determined by agar dilution method. One-way Analysis of Variance (ANOVA followed by Sidak post hoc test was used to compare the mean zone of microbial growth in the groups.Results: There were significant differences between the two groups (p< 0.05. Results of the agar diffusion test showed that antibiotics (amoxicillin, gentamycin had the greatest antibacterial activity followed by NSAIDs (ibuprofen, diclofenac. Ca(OH2 failed to show antibacterial activity. Diclofenac and ibuprofen showed distinct antibacterial activity against E. faecalis in 50 µg/ml and above concentrations.Conclusion: Within the limitations of this in vitro study, it is concluded that diclofenac and ibuprofen have significantly more pronounced antibacterial activity against E. faecalis in comparison with Ca(OH2.
Moore, John E; Gilpin, Deidre; Crothers, Elizabeth; Canney, Anne; Kaneko, Aki; Matsuda, Motoo
An investigation was carried out into the prevalence of thermophilic Campylobacter subspecies (spp.) and Cryptosporidium spp. in fresh fecal specimens collected from members of the gull family (Larus spp.) from three coastal locations of Northern Ireland. A total of 205 fresh fecal specimens were collected from gulls, of which 28 of 205 (13.7%) were positive for Campylobacter spp. and none of 205 for Cryptosporidium spp. Of these campylobacters, 21 of 28 (75%) isolates obtained belonged to the urease-positive thermophilic Campylobacter (UPTC) taxon, followed by five of 28 (17.9%) Campylobacter lari and 2/28 (7.1%) Campylobacter jejuni. It is significant that seagulls are the sole warm-blooded animal host of this bacterial taxon in Northern Ireland. It is proposed that physiological adaptation to starvation by gulls may lead to increased concentrations of urea through energy production from protein, yielding increased levels of urea for metabolism by UPTC organisms. In general, the possibility exists that environmental contamination of surface waters with campylobacters might be mediated by wild birds (such as gulls), where such waters are used for recreational purposes or where such waters are consumed untreated, might represent a risk to public health.
Full Text Available The aim of this study is to identify and monitor the presence of Aeromonas spp. strains in stool cultures. We analyzed 5564 stool cultures from September 2012 to August 2013. Sixty-three patients were positive for Aeromonas spp. The most frequent symptoms were: diarrhea (46.0% and abdominal pain (12.7%. Pediatric subjects were 28. Samples’ microscopic examination showed leukocytes in 38.1% of cases. It is still controversial whether Aeromonas are responsible for human gastroenteritis, but their presence in faecies of symptomatic patients supports their etiologic role. We propose search for toxins by polymerase chain reaction to identify strains that require an antibiotic therapy.
Vandera, Elpiniki; Tsirka, Georgia; Kakouri, Athanasia; Koukkou, Anna-Irini; Samelis, John
Enterococci are naturally selected for growth in thermized ewes'/goats' milk mixtures used for traditional cooked hard cheese processing in Greece. A culture-independent PCR-based approach was applied to detect the presence of enterocin-encoding genes in naturally culture-enriched thermized milk (TM). Portions of TM (63 °C, 30 s) collected from a commercial cheese plant before addition of starters were fermented at 37 °C for 48 h to facilitate growth of indigenous enterococci. The multiple enterocin-producing (m-Ent+) Enterococcus faecium KE82 and the nisin A-producing Lactococcus lactis subsp. cremoris M104 served as bacteriocin-positive inocula in separate TM treatments. The PCR results revealed a constant presence of the enterocin A, B and P genes in TM fermented naturally at 37 °C. Eleven out of 42 (26.2%) lactic isolates from the enriched TM cultures without inoculation were Ent+ E. faecium assigned to three biotypes. Biotype I (4 isolates) included single entA possessors, whereas biotype II (5 isolates) and biotype III (2 isolates) were m-Ent+ variants profiling entA-entB-entP and entA-entB genes, respectively. Biotype II displayed the strongest antilisterial activity in vitro. Surprisingly, 85.7% (6/7) of the m-Ent+ E. faecium were selectively isolated from Baird-Parker agar, reflecting their natural resistance to 0.01% tellurite contained in the egg yolk supplement. No cytolysin-positive E. faecalis or other Ent+ Enterococcus spp. were isolated. In conclusion, commercially thermized Greek milk is a natural pool or 'reservoir' of antagonistic Ent+ or m-Ent+ E. faecium strains that can be easily detected and recovered by applying this PCR-based approach to naturally fermented milks or cheese products. Copyright © 2018 Elsevier B.V. All rights reserved.
Abbaszadegan, Abbas; Sahebi, Safoora; Gholami, Ahmad; Delroba, Alireza; Kiani, Amin; Iraji, Aida; Abbott, Paul Vincent
In the present in vitro study, we investigated the time-related antimicrobial efficacy of Aloe vera and Zataria multiflora (Z. multiflora) plant essential oils compared to calcium hydroxide ([Ca[OH]2 ) to eliminate Enterococcus faecalis (E. faecalis) from root canals. A new strain of E. faecalis (Enterococcus spp. AGH04) was isolated from a previously root-filled tooth with persistent apical periodontitis. The 16S rRNA sequence was analyzed and deposited in GeneBank under accession number KF465681. A total of 108 extracted human single-rooted teeth were contaminated with this bacterial strain and treated with Aloe vera essential oil, Z. multiflora essential oil, and Ca(OH)2 for 1, 7, and 14 days. Gas chromatography-mass spectrometry (GC-MS) was used to determine the chemical composition of the oils. The percentage reduction from initial c.f.u./mL counts were calculated and analyzed. Carvacrol, thymol, and linalool were the main constituents of both essential oils. The c.f.u./mL count reductions significantly increased for all three medicaments when the contact time was extended. A statistically-significant difference was observed between the medicaments after 1 and 7 days, but there was no significant difference after 14 days. Both medicinal herbs showed equal antimicrobial efficiency against E. faecalis, comparable to Ca(OH)2 for the prolonged contact time of 14 days. © 2014 Wiley Publishing Asia Pty Ltd.
Jung, Arne; Metzner, Martin; Köhler-Repp, Dagmar; Rautenschlein, Silke
Enterococcus cecorum (EC) was thus far only known as a pathogen for broilers and broiler breeders. Recently there was evidence of EC field outbreaks in Pekin duck flocks in Germany. In this study we experimentally reproduced an EC infection in Pekin ducks. At 12 days post hatch, groups of Pekin ducks were infected orally, via the thoracic air sac or intravenously with 1.5 × 10(9) colony-forming units (CFU) of EC per bird or via the air sac with 8.5 × 10(5) or 8.5 × 10(7) CFU per bird. Ducks of the intravenously infected group showed 100% mortality after 2 days post infection. The air sac inoculated high-dose group exhibited a mortality rate of 67%. Birds that were infected with 8.5 × 10(5) and 8.5 × 10(7) CFU showed 6.7% mortality after 7 days post infection. Dead birds displayed pneumonia, airsacculitis, pericarditis and splenitis and EC was re-isolated from these organs. Surviving birds of all groups apart from the orally infected ducks demonstrated clinical signs such as huddling, reduced mobility and diarrhoea. Furthermore, they showed gross pathological lesions including airsacculitis and splenitis and lower bodyweights than the control group at necropsy on days 7, 14 and 21 post infection. The present study clearly confirms that EC is pathogenic for Pekin ducks after experimental infection via the intravenous route or the respiratory tract. EC therefore has to be considered as an emerging avian pathogen not only in broilers but also in Pekin ducks.
Full Text Available Enterococcus faecalis is an opportunistic nosocomial pathogen that is highly resistant to a variety of environmental insults, including an intrinsic tolerance to antimicrobials that target the cell wall (CW. With the goal of determining the CW-stress stimulon of E. faecalis, the global transcriptional profile of E. faecalis OG1RF exposed to ampicillin, bacitracin, cephalotin or vancomycin was obtained via microarrays. Exposure to the β-lactams ampicillin and cephalotin resulted in the fewest transcriptional changes with 50 and 192 genes differentially expressed 60 min after treatment, respectively. On the other hand, treatment with bacitracin or vancomycin for 60 min affected the expression of, respectively, 377 and 297 genes. Despite the differences in the total number of genes affected, all antibiotics induced a very similar gene expression pattern with an overrepresentation of genes encoding hypothetical proteins, followed by genes encoding proteins associated with cell envelope metabolism as well as transport and binding proteins. In particular, all drug treatments, most notably bacitracin and vancomycin, resulted in an apparent metabolic downshift based on the repression of genes involved in translation, energy metabolism, transport and binding. Only 19 genes were up-regulated by all conditions at both the 30 and 60 min time points. Among those 19 genes, 4 genes encoding hypothetical proteins (EF0026, EF0797, EF1533 and EF3245 were inactivated and the respective mutant strains characterized in relation to antibiotic tolerance and virulence in the Galleria mellonella model. The phenotypes obtained for two of these mutants, ΔEF1533 and ΔEF3245, support further characterization of these genes as potential candidates for the development of novel preventive or therapeutic approaches.
Eijsink Vincent GH
Full Text Available Abstract Background Surface proteins are a key to a deeper understanding of the behaviour of Gram-positive bacteria interacting with the human gastro-intestinal tract. Such proteins contribute to cell wall synthesis and maintenance and are important for interactions between the bacterial cell and the human host. Since they are exposed and may play roles in pathogenicity, surface proteins are interesting targets for drug design. Results Using methods based on proteolytic "shaving" of bacterial cells and subsequent mass spectrometry-based protein identification, we have identified surface-located proteins in Enterococcus faecalis V583. In total 69 unique proteins were identified, few of which have been identified and characterized previously. 33 of these proteins are predicted to be cytoplasmic, whereas the other 36 are predicted to have surface locations (31 or to be secreted (5. Lipid-anchored proteins were the most dominant among the identified surface proteins. The seemingly most abundant surface proteins included a membrane protein with a potentially shedded extracellular sulfatase domain that could act on the sulfate groups in mucin and a lipid-anchored fumarate reductase that could contribute to generation of reactive oxygen species. Conclusions The present proteome analysis gives an experimental impression of the protein landscape on the cell surface of the pathogenic bacterium E. faecalis. The 36 identified secreted (5 and surface (31 proteins included several proteins involved in cell wall synthesis, pheromone-regulated processes, and transport of solutes, as well as proteins with unknown function. These proteins stand out as interesting targets for further investigation of the interaction between E. faecalis and its environment.
Hammerum, Anette M; Baig, Sharmin; Kamel, Yasmin
Objectives: To describe the changing epidemiology of vancomycin-resistant Enterococcus faecium and Enterococcus faecalis in clinical samples in Denmark 2005-15 according to species and van type, and, furthermore, to investigate the genetic relatedness of the clinical E. faecium isolates from 2015...... were detected along with 1 vanA E. faecalis and 1 vanB E. faecalis . cgMLST subdivided the 368 vanA E. faecium isolates into 33 cluster types (CTs), whereas the vanB E. faecium isolate belonged to a different CT. ST203-CT859 was most prevalent (51%), followed by ST80-CT14 (22%), ST117-CT24 (6%), ST80...
Falkinham III, Joseph O.; Williams, Myra D.; Kwait, Rebecca; Lande, Leah
Objective/Background: A published survey of bacteria in showerhead biofilm samples revealed that Methylobacterium spp. and Mycobacterium spp. seldom coexisted in biofilms. Method: To confirm that information, biofilm samples were collected from household plumbing of Mycobacterium avium patients and Methylobacterium spp. and M. avium numbers were measured by direct colony counts. Results: The results demonstrated that if Methylobacterium spp. were present, Mycobacterium spp. were absent,...
Ueno, Hiroshi; Minagawa, Yoshihiro; Hara, Mayu; Rahman, Suhaila; Yamato, Ichiro; Muneyuki, Eiro; Noji, Hiroyuki; Murata, Takeshi; Iino, Ryota
V-ATPase (VoV1) converts the chemical free energy of ATP into an ion-motive force across the cell membrane via mechanical rotation. This energy conversion requires proper interactions between the rotor and stator in VoV1 for tight coupling among chemical reaction, torque generation, and ion transport. We developed an Escherichia coli expression system for Enterococcus hirae VoV1 (EhVoV1) and established a single-molecule rotation assay to measure the torque generated. Recombinant and native EhVoV1 exhibited almost identical dependence of ATP hydrolysis activity on sodium ion and ATP concentrations, indicating their functional equivalence. In a single-molecule rotation assay with a low load probe at high ATP concentration, EhVoV1 only showed the “clear” state without apparent backward steps, whereas EhV1 showed two states, “clear” and “unclear.” Furthermore, EhVoV1 showed slower rotation than EhV1 without the three distinct pauses separated by 120° that were observed in EhV1. When using a large probe, EhVoV1 showed faster rotation than EhV1, and the torque of EhVoV1 estimated from the continuous rotation was nearly double that of EhV1. On the other hand, stepping torque of EhV1 in the clear state was comparable with that of EhVoV1. These results indicate that rotor-stator interactions of the Vo moiety and/or sodium ion transport limit the rotation driven by the V1 moiety, and the rotor-stator interactions in EhVoV1 are stabilized by two peripheral stalks to generate a larger torque than that of isolated EhV1. However, the torque value was substantially lower than that of other rotary ATPases, implying the low energy conversion efficiency of EhVoV1. PMID:25258315
E. A. Synetar
Full Text Available The nature of microorganisms can exist in two physiological forms that allow microbes to preserve livelihoods and continue their life cycle. The first is the population of planktonic forms of microorganisms which live freely in the environment with the developed systems of active and passive mobility, contributing to the rapid spread of a liquid medium. The second forms are those expressing specific mechanisms of adhesion, and able to aggregate on biogenic and abiogenic surfaces. Even in the deep sea vast number of species of bacteria live in their inherent horizons. Thus, the study of biofilms tube life support systems, diagnostic, laparoscopic devices during prolonged catheterization of the urinary system is of great practical, theoretical and biological significance in medicine and biology. For almost 20% of catheter-associated infections antibiotic therapy is uneffective, particularly through the formation of microbial biofilms on the surface of urinary catheters. We characterized the dynamics of biofilm growth of Enterococcus faecalis on fragments ofsilicone catheter. The study was conducted using bacteriological and electron microscopic techniques. Study of the dynamics of biofilm formation was performed using E. faecalis strain 49, which is isolated from the urine of persons who are not the patients of the urological department of resuscitation and intensive therapy. Using scanning electron microscopy we have established dynamics and phase attachment ofE. faecalis bacteria and subsequent overgrowth of silicone catheter surface. Aftercalculations, index of adhesion on the turbulent wall amounted to 0,49 microbial cells. That is, every other cell of the monolayer adhered on the catheter. Area of biofilm growth of E. faecalis after 24 hour incubation was equal to 51.5 μm2, in 48 hours it increased to 231.5 μm2. After 72 hours of incubation we recorded the increase in biofilm growth of E. faecalisto 1922,8 μm2. The results were obtained
Armianty Armianty; Indrya Kirana Mattulada
Enterococcus faecalis is one of bacteria which have resistance against the pulp tissue defense mechanism and often found in endodontic infections. This bacterial is may hold good colonization, can survive in the root canal without other bacteria, and is capable of producing toxins directly or through the induction of inflammation. Betel leaf (Piper betle Linn)contain essential oils, as the main components of the essential oils are phenols and compounds, among other derivate such a...
Ladero, Victor; Linares, Daniel M.; del Rio, Beatriz; Fernandez, Maria; Martin, M. Cruz
We here report a 3.059-Mbp draft assembly for the genome of Enterococcus durans strain IPLA 655. This dairy isolate provides a model for studying the regulation of the biosynthesis of tyramine (a toxic compound). These results should aid our understanding of tyramine production and allow tyramine accumulation in food to be reduced. PMID:23682153
Menon, Vidthiya; Davis, Rebecca; Shackel, Nick; Espedido, Bjorn A; Beukers, Alicia G; Jensen, Slade O; van Hal, Sebastiaan J
Daptomycin β-Lactam combination therapy offers "protection" against daptomycin non-susceptibility (DNS) development in Enterococcus faecium. We report failure of this strategy and the importance of source control. Mutations were detected in the LiaF and cls genes in DNS isolates. A single DNS isolate contained an unrecognized mutation, which requires confirmation. Copyright © 2017 Elsevier Inc. All rights reserved.
Seedat, Jamela; Zick, Günther; Klare, Ingo; Konstabel, Carola; Weiler, Norbert; Sahly, Hany
We report the emergence of linezolid resistance (MICs of 16 to 32 mg/liter) in clonally related vancomycin-susceptible and -resistant Enterococcus faecium isolates from an intensive care unit patient after 12 days of linezolid therapy. Only linezolid-susceptible isolates of the same clone were detected at 28 days after termination of linezolid therapy.
Acharya, A; Khanal, A; Kanungo, R; Mohapatra, T
Life threatening infections caused by enterococcus species with multidrug resistance has emerged as a threat to medical care in the present era. This study was conducted to characterize enterococcus species isolated from different clinical samples and to detect the pattern of susceptibility to some of the commonly used antibiotics in B.P Koirala Institute of Health Sciences (BPKIHS), a tertiary care hospital in eastern Nepal. Clinical samples submitted to the microbiology unit of Central Laboratory Service (CLS) for culture and sensitivity during March 2002 - February 2003 was analyzed. Enterococcus species were identified by colony characteristics, gram staining and relevant biochemical tests. Antibiotic susceptibility test was done by the Kirby Bauer disc diffusion technique. Of 50 Enterococcus species isolated, E. faecalis was the predominant isolate (48.0%) followed by E. faecium (32.0%) and E. avium (20.0%). Eighty-eight percent of E. faecalis showed sensitivity to cephotaxime and 87.0% to vancomycin. Multiple drug resistance was observed most commonly in E. faecium. Seventeen percent of E. faecium were resistant to vancomycin and 63.0% to ciprofloxacin and 44.0% to ampicillin. On the contrary E. avium rarely showed resistance to the antimicrobials tested including vancomycin. Enterococcal infections are common nowadays specially in hospitalized patients. Inappropriate use of antibiotics in clinical practice and poultry should be discouraged to prevent the emergence of multidrug resistant species.
Chang, S-Y; Chen, Y-S; Pan, S-F; Lee, Y-S; Chang, C-H; Chang, C-H; Yu, B; Wu, H-C
Purification and characterization of a novel bacteriocin produced by strain Enterococcus faecium D081821. Enterococcus faecium D081821, isolated from the traditional Taiwanese fermented food dochi (fermented black beans), was previously found to produce a bacteriocin against Listeria monocytogenes and some Gram-positive bacteria. This bacteriocin, termed enterocin TW21, was purified from culture supernatant by ammonium sulfate precipitation, Sep-Pak C18 cartridge, ion-exchange and gel filtration chromatography. Mass spectrometry analysis showed the mass of the peptide to be approximately 5300·6 Da. The N-terminal amino acid sequencing yielded a partial sequence NH2 -ATYYGNGVYxNTQK by Edman degradation, and it contains the consensus class IIa bacteriocin motif YGNGV in the N-terminal region. The open reading frame (ORF) encoding the bacteriocin was identified from the draft genome sequence of Enterococcus faecium D081821, and sequence analysis of this peptide indicated that enterocin TW21 is a novel bacteriocin. Enterococcus faecium D081821 produced a bacteriocin named enterocin TW21, the molecular weight and amino acid sequence both revealed it to be a novel bacteriocin. A new member of class IIa bacteriocin was identified. This bacteriocin shows great inhibitory ability against L. monocytogenes and could be applied as a natural food preservative. © 2013 The Society for Applied Microbiology.
Perez, Marta; Ladero, Victor; del Rio, Beatriz; Redruello, Begona; de Jong, Anne; Kuipers, Oscar; Kok, Jan; Martin, M. Cruz; Fernandez, Maria; Alvarez, Miguel A.
Enterococci are considered mainly responsible for the undesirable accumulation of the biogenic amines tyramine and putrescine in cheeses. The biosynthesis of tyramine and putrescine has been described as a species trait in Enterococcus faecalis. Tyramine is formed by the decarboxylation of the amino
van der Waal, S.V.; Jiang, L.M.; de Soet, J.J.; van der Sluis, L.W.M.; Wesselink, P.R.; Crielaard, W.
Incomplete disinfection of the root canal system is a major cause of post-treatment disease. This study aimed to investigate the disinfecting property of organic acid salts and sodium chloride (NaCl), in a double-hurdle strategy, on Enterococcus faecalis biofilms. First of all, the high-throughput
van der Waal, Suzette V.; Jiang, Lei-Meng; de Soet, Johannes J.; van der Sluis, Lucas W. M.; Wesselink, Paul R.; Crielaard, Wim
Incomplete disinfection of the root canal system is a major cause of post-treatment disease. This study aimed to investigate the disinfecting property of organic acid salts and sodium chloride (NaCl), in a double-hurdle strategy, on Enterococcus faecalis biofilms. First of all, the high-throughput
de Been, Mark; Pinholt, Mette; Top, Janetta
Enterococcus faecium, a common inhabitant of the human gut, has emerged as an important multidrug-resistant nosocomial pathogen in the last two decades. Since the start of the 21(st) century, multi-locus sequence typing (MLST) has been used to study the molecular epidemiology of E. faecium. However...
C. Abrantes, Marta; Kok, Jan; de Fatima Silva Lopes, Maria
Two Enterococcus faecalis genes encoding the P-type ATPase EF1400 and the putative SapB protein EF0759 were previously shown to be strongly upregulated in the presence of high concentrations of zinc. In the present work, we showed that a Zn(2+)-responsive DNA-binding motif (zim) is present in the
Wilcks, Andrea; Andersen, Sigrid Rita; Licht, Tine Rask
The prevalence of tetracycline resistance, and of specific genetic determinants for this resistance was investigated in 1003 strains of Enterococcus faecalis isolated from various raw food products originating from five categories including chicken meat, other poultry meat, beef, pork, and 'other...
Tytgat, Hanne L.P.; Douillard, François P.; Reunanen, Justus; Rasinkangas, Pia; Hendrickx, Antoni P.A.; Laine, Pia K.; Paulin, Lars; Satokari, Reetta; Vos, de Willem M.
Vancomycin-resistant enterococci (VRE) have become a major nosocomial threat. Enterococcus faecium is of special concern, as it can easily acquire new antibiotic resistances and is an excellent colonizer of the human intestinal tract. Several clinical studies have explored the potential use of
Leendertse, Masja; Heikens, Esther; Wijnands, Lucas M.; van Luit-Asbroek, Miranda; Teske, Gwendoline J. D.; Roelofs, Joris J. T. H.; Bonten, Marc J. M.; van der Poll, Tom; Willems, Rob J. L.
The role that the enterococcal surface protein Esp plays in the capacity of Enterococcus faecium to adhere to uroepithelial cells and the role that it plays in urinary tract infection and peritonitis was investigated in vitro and in vivo, respectively, using Esp-expressing E. faecium (E1162) and its
The objective of this study was to compare dependency of survival rates on temperature for indicator organisms E. coli and Enterococcus and the pathogen Salmonella in surface waters. A database of 86 survival datasets from peer-reviewed papers on inactivation of E. coli, Salmonel...
Brede, Dag Anders; Snipen, Lars Gustav; Ussery, David
The genome of Enterococcus faecalis 62, a commensal isolate from a healthy Norwegian infant, revealed multiple adaptive traits to the gastrointestinal tract (GIT) environment and the milk-containing diet of breast-fed infants. Adaptation to a commensal existence was emphasized by lactose and other...
For many years Enterococcus faecium was considered a commensal of the digestive tract, which only sporadically caused opportunistic infections in severely ill patients. Over the last two decades, vancomycin resistant E. faecium (VREF) has emerged worldwide as an important cause of nosocomial
JiaJi Qiu; Jie Tang; DeLing Li
Background:Vancomycin-resistant Enterococcus faecium ventriculitis is one of the most severe events in postneurosurgical intracranial infections.There are no guidelines recommending an appropriate treatment before.Case presentation:This case presents a successful linezolid treatment for post-neurosurgical vancomycin-resistant Enterococcus faecium ventriculitis of a 24-year-old man in the department of neurosurgery,Beijing Tiantan Hospital.Conclusions:Linezolid should be considered as one of the important methods for the treatment of postneurosurgical intracranial infections caused by vancomycin-resistant Enterococcus.
Larissa Sbaglia Celiberto
Full Text Available Some probiotic strains have the potential to assist in relieving the symptoms of inflammatory bowel disease. The impact of daily ingestion of a soy-based product fermented by Enterococcus faecium CRL 183 and Lactobacillus helveticus 416 with the addition of Bifidobacterium longum ATCC 15707 on chemically induced colitis has been investigated thereof within a period of 30 days.Colitis was induced by dextran sulfate sodium. The animals were randomly assigned into five groups: Group C: negative control; Group CL: positive control; Group CLF: DSS with the fermented product; Group CLP: DSS with the non-fermented product (placebo; Group CLS: DSS with sulfasalazine. The following parameters were monitored: disease activity index, fecal microbial analyses, gastrointestinal survival of probiotic microorganisms and short-chain fatty acids concentration in the feces. At the end of the protocol the animals' colons were removed so as to conduct a macroscopical and histopathological analysis, cytokines and nitrite quantification.Animals belonging to the CLF group showed fewer symptoms of colitis during the induction period and a lower degree of inflammation and ulceration in their colon compared to the CL, CLS and CLP groups (p<0.05. The colon of the animals in groups CL and CLS presented severe crypt damage, which was absent in CLF and CLP groups. A significant increase in the population of Lactobacillus spp. and Bifidobacterium spp. at the end of the protocol was verified only in the CLF animals (p<0.05. This group also showed an increase in short-chain fatty acids (propionate and acetate. Furthermore, the intestinal survival of E. faecium CRL 183 and B. longum ATCC 15707 in the CLF group has been confirmed by biochemical and molecular analyzes.The obtained results suggest that a regular intake of the probiotic product, and placebo to a lesser extent, can reduce the severity of DSS-induced colitis on rats.
Chang, S; Kang, D-H
To develop a new isolation medium with higher recovery rates of Alicyclobacillus spp. SK agar was developed with optimized incubation temperature, pH, acidulant, Tween 80 concentration and divalent cation addition. Results indicate that detection of Alicyclobacillus spp. by SK agar was significantly higher (P > 0.05) than those obtained by K agar, orange serum agar, and potato dextrose agar. Current media used for Alicyclobacillus spp. isolation still resulted in high numbers of false negative products. The sensitivity of SK agar to Alicyclobacillus spp. allows detection of low numbers of Alicyclobacillus spp. and also provides a more higher isolation results compared with currently used media. SK agar will be useful to the fruit juice industry to obtain more accurate numbers of contaminant Alicyclobacillus spp. With this media, false negative samples can be reduced, and the likelihood of exported products being rejected can be greatly reduced.
Zhao, Y.; Aarnink, A.J.A.; Xin, H.
High microbial concentrations and emissions associated with livestock houses raise health and environmental concerns. A pilot-scale ultraviolet photocatalytic (UV-PCO) scrubber was tested for its efficacy to inactivate aerosolized Enterococcus faecalis and infectious bursal disease virus (IBDV).
Enterococcus cecorum has been implicated as a possible cause of disease in poultry including spondylitis, vertebral osteoarthritis and femoral osteomyelitis. Antimicrobials have been used to treat the disease during outbreaks, but have been ineffective in controlling outbreak mortality possibly due...
Full Text Available Sara Soheili,1 Sobhan Ghafourian,2 Zamberi Sekawi,1 Vasantha Kumari Neela,1 Nourkhoda Sadeghifard,2 Morovat Taherikalani,2 Afra Khosravi,2 Ramliza Ramli,3 Rukman Awang Hamat11Department of Medical Microbiology and Parasitology, Faculty of Medicine and Health sciences, Universiti Putra Malaysia, Serdang, Malaysia; 2Clinical Microbiology Research Center, Ilam University of Medical Sciences, Ilam, Iran; 3Department of Medical Microbiology and Immunology, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaakob Latif, Bandar Tun Razak, Kuala Lumpur, Malaysia Abstract: The toxin–antitoxin (TA system is a regulatory system where two sets of genes encode the toxin and its corresponding antitoxin. In this study, the prevalence of TA systems in independently isolated clinical isolates of Enterococcus faecium and Enterococcus faecalis was determined, the dominant TA system was identified, different virulence genes in E. faecium and E. faecalis were surveyed, the level of expression of the virulence and TA genes in normal and stress conditions was determined, and finally their associations with the TA genes were defined. Remarkably, the analysis demonstrated higBA and mazEF in all clinical isolates, and their locations were on chromosomes and plasmids, respectively. On the other hand, a quantitative analysis of TA and virulence genes revealed that the expression level in both genes is different under normal and stress conditions. The results obtained by anti-mazF peptide nucleic acids demonstrated that the expression level of virulence genes had decreased. These findings demonstrate an association between TA systems and virulence factors. The mazEF on the plasmids and the higBA TA genes on the chromosomes of all E. faecium and E. faecalis strains were dominant. Additionally, there was a decrease in the expression of virulence genes in the presence of anti-mazF peptide nucleic acids. Therefore, it is suggested that mazEF TA systems
Sharareh Mousavi Zahed
Full Text Available Background and Aims: To ensure root canal treatment success, endodontic microbiota should be efficiently reduced. Several irrigation devices have been recently introduced with the main objective of improving root canal disinfection.The purpose of this study was to evaluate the rinsing effect of RinsEndo system in reduction of enterococcus faecalis in comparison with conventional hand syringe in infected root canals. Materials and Methods: 60 extracted single canal anterior teeth were infected with enterococcus faecalis and divided into 3 groups: RinsEndo system, conventional hand syringe and control group. The enterococcus faecalis colonies were counted in each group before and after rinsing. Data were analyzed using Variance and Kruskal Wallis test. Results: The mean of enterococcus faecalis growth after rinsing was 3.50×103 in group with conventional syring rinsing, 2.04×103 in group with RinsEndo washing and 6.11×103 in control group. Reduction of enterococcus faecalis after rinsing was statistically significant in each group (P<0.001. The amount of reduction in number of colonies with RinsEndo and conventional syringe rinsing was higher in comparison with control group and this difference was significant (P<0.001. RinsEndo rinsing effect was statistically significantly higher in comparison to conventional syringe as well (P<0.001. Conclusion: Rinsing with RinsEndo system was significantly more efficient in reduction of enterococcus faecalis from root canal in comparison with hand syringe washing.
Chen, Yi-Sheng; Yu, Chi-Rong; Ji, Si-Hua; Liou, Min-Shiuan; Leong, Kun-Hon; Pan, Shwu-Fen; Wu, Hui-Chung; Lin, Yu-Hsuan; Yu, Bi; Yanagida, Fujitoshi
Enterococcus sp. 812, isolated from fresh broccoli, was previously found to produce a bacteriocin active against a number of Gram-positive bacteria, including Listeria monocytogenes. Bacteriocin activity decreased slightly after autoclaving (121 °C for 15 min), but was inactivated by protease K. Mass spectrometry analysis revealed the bacteriocin mass to be approximately 4,521.34 Da. N-terminal amino acid sequencing yielded a partial sequence, NH2-ATYYGNGVYXDKKKXWVEWGQA, by Edman degradation, which contained the consensus class IIa bacteriocin motif YGNGV in the N-terminal region. The obtained partial sequence showed high homology with some enterococcal bacteriocins; however, no identical peptide or protein was found. This peptide was therefore considered to be a novel bacteriocin produced by Enterococcus sp. 812 and was termed enterocin T.
Thamsborg, Stig M.; Ketzis, Jennifer; Horii, Yoichiro
This paper reviews the occurrence and impact of threadworms, Strongyloides spp., in companion animals and large livestock, the potential zoonotic implications and future research. Strongyloides spp. infect a range of domestic animal species worldwide and clinical disease is most often encountered...
Nešuta, Ondřej; Monincová, Lenka; Buděšínský, Miloš; Hexnerová, Rozálie; Hadravová, Romana; Čeřovský, Václav
Roč. 15, č. 1 (2017), s. 24-25 ISSN 2336-7202. [Mezioborové setkání mladých biologů, biochemiků a chemiků /17./. 30.05.2017-01.06.2017, Milovy] R&D Projects: GA TA ČR(CZ) TA04010638; GA MZd(CZ) NV16-27726A Institutional support: RVO:61388963 Keywords : antimicrobial peptides * bacterial resistance * Enterococcus faecalis Subject RIV: EE - Microbiology, Virology
Oh, Won Sup; Ko, Kwan Soo; Song, Jae-Hoon; Lee, Mi Young; Park, Sulhee; Peck, Kyong Ran; Lee, Nam Yong; Kim, Choon-Kwan; Lee, Hyuck; Kim, Shin-Woo; Chang, Hyun-Ha; Kim, Yeon-Sook; Jung, Sook-In; Son, Jun Seong; Yeom, Joon-Sup; Ki, Hyun Kyun; Woo, Gun-Jo
We tested the in vitro susceptibilities of 603 enterococcal isolates from eight tertiary-care hospitals in Korea. The quinupristin-dalfopristin resistance rate in Enterococcus faecium was very high (25 isolates, 10.0%). It was suggested that both clonal spread and the sporadic emergence of quinupristin-dalfopristin-resistant isolates may explain the high prevalence of quinupristin-dalfopristin resistance in Korea. PMID:16304198
Eckert , Catherine; Lecerf , Maxime; Dubost , Lionel; Arthur , Michel; Mesnage , Stéphane
International audience; The major peptidoglycan hydrolase of Enterococcus faecalis, AtlA, has been identified, but its enzyme activity remains unknown. We have used tandem mass spectrometry analysis of peptidoglycan hydrolysis products obtained using the purified protein to show that AtlA is an N-acetylglucosaminidase. To gain insight into the regulation of its enzyme activity, the three domains of AtlA were purified alone or in combination following expression of truncated forms of the atlA ...
Vatkar, Niranjan Ashok; Hegde, Vivek; Sathe, Sucheta
Aim: To compare the vitality of Enterococcus faecalis within dentinal tubules after subjected to five root canal disinfection methods. Materials and Methods: Dentin blocks (n = 60) were colonized with E. faecalis. After 4 weeks of incubation, the dentin blocks were divided into one control and five test groups (n = 10 each). The root canals of test groups were subjected to one of the disinfection methods, namely, normal saline (NS), sodium hypochlorite (NaOCl), chlorhexidine digluconate (C...
Full Text Available Orbital cellulitis is an infection of soft tissue behind the orbital septum. Common pathogens isolated include Staphylococcus aureus, Staphylococcus epidermidis, and Streptococcus pneumoniae. It is a straightforward diagnosis and usually responds to empirical treatment without any sequela. We report a case of orbital cellulitis caused by Enterococcus faecalis, which was complicated by myositis of levator palpebrae superioris. To the best of our knowledge, only one case report exists dating way back to 1986.
Fritzenwanker, Moritz; Chakraborty, Anindita; Hain, Torsten; Zimmermann, Kurt; Domann, Eugen
The probiotic Symbioflor 1 is a historical concoction of 10 isolates of Enterococcus faecalis. Pulsed-field gel electrophoresis revealed two groups: one comprising eight identical clones (DSM16430, DSM16432, DSM16433, DSM16435 to DSM16439) and a further two isolates (DSM16431, DSM16434) with marginally different profiles. Here, we report a comparative analysis of the draft genome sequences of representative isolates.
Herranz, Carmen; Cintas, Luis M.; Hernández, Pablo E.; Moll, Gert N.; Driessen, Arnold J. M.
Enterocin P is a bacteriocin produced by Enterococcus faecium P13. We studied the mechanism of its bactericidal action using enterocin-P-sensitive E. faecium T136 cells. The bacteriocin is incapable of dissipating the transmembrane pH gradient. On the other hand, depending on the buffer used, enterocin P dissipates the transmembrane potential. Enterocin P efficiently elicits efflux of potassium ions, but not of intracellularly accumulated anions like phosphate and glutamate. Taken together, t...
Huang, En; Zhang, Liwen; Chung, Yoon-Kyung; Zheng, Zuoxing; Yousef, Ahmed E.
Use of bacteriocins in food preservation has received great attention in recent years. The goal of this study is to characterize enterocin RM6 from Enterococcus faecalis OSY-RM6 and investigate its efficacy against Listeria monocytogenes in cottage cheese. Enterocin RM6 was purified from E. faecalis culture supernatant using ion exchange column, multiple C18-silica cartridges, followed by reverse-phase high-performance liquid chromatography. The molecular weight of enterocin RM6 is 7145.0823 ...
Driessen, A J; Smid, E J; Konings, W N
Enterococcus faecalis ATCC 11700 is able to use arginine and the diamine agmatine as a sole energy source. Via the highly homologous deiminase pathways, arginine and agmatine are converted into CO2, NH3, and the end products ornithine and putrescine, respectively. In the arginine deiminase pathway, uptake of arginine and excretion of ornithine are mediated by an arginine-ornithine antiport system. The translocation of agmatine was studied in whole cells grown in the presence of arginine, agma...
Velraeds, M M; van der Mei, H C; Reid, G; Busscher, H J
In this study, 15 Lactobacillus isolates were found to produce biosurfactants in the mid-exponential and stationary growth phases. The stationary-phase biosurfactants from lactobacillus casei subsp. rhamnosus 36 and ATCC 7469, Lactobacillus fermentum B54, and Lactobacillus acidophilus RC14 were investigated further to determine their capacity to inhibit the initial adhesion of uropathogenic Enterococcus faecalis 1131 to glass in a parallel-plate flow chamber. The initial deposition rate of E....
Combined effects of extremely high frequency electromagnetic field and antibiotics on Enterococcus hirae ATCC 9790 bacterial growth and survival were investigated using 51.8 GHz and 53 GHz frequencies in combination with two commonly used antibiotics: ampicillin and dalacin. Results revealed that, despite bacterial type and membrane structure and properties, the combined effect, especially with 53 GHz and dalacin, suppresses bacterial growth and decreases their survival
Hodges, T L; Zighelboim-Daum, S; Eliopoulos, G M; Wennersten, C; Moellering, R C
Penicillin-"virgin" strains of Enterococcus faecalis collected from a population of individuals with no previous antibiotic exposure were subjected in vitro to penicillin delivered as repeated pulses, stepwise increasing concentrations, or sustained levels of a single concentration. Changes in resistance to penicillin were assessed by determination of MICs, and time-kill studies were performed to evaluate changes in tolerance to the bactericidal effects of penicillin. Isogenic clones, derived...
Green, M; Barbadora, K; Wadowsky, R M
The combination of ampicillin and vancomycin kills some but not all strains of ampicillin- and vancomycin-resistant Enterococcus faecium. We compared a simple test for synergy utilizing a commercially available microdilution susceptibility system with time-kill studies and determined acceptable breakpoints for this test for 20 strains of ampicillin- and vancomycin-resistant E. faecium. The combination of ampicillin and vancomycin was tested for synergy by time-kill, broth macrodilution, and b...
Soheili, Sara; Ghafourian, Sobhan; Sekawi, Zamberi; Neela, Vasantha Kumari; Sadeghifard, Nourkhoda; Taherikalani, Morovat; Khosravi, Afra; Ramli, Ramliza; Hamat, Rukman Awang
The toxin-antitoxin (TA) system is a regulatory system where two sets of genes encode the toxin and its corresponding antitoxin. In this study, the prevalence of TA systems in independently isolated clinical isolates of Enterococcus faecium and Enterococcus faecalis was determined, the dominant TA system was identified, different virulence genes in E. faecium and E. faecalis were surveyed, the level of expression of the virulence and TA genes in normal and stress conditions was determined, and finally their associations with the TA genes were defined. Remarkably, the analysis demonstrated higBA and mazEF in all clinical isolates, and their locations were on chromosomes and plasmids, respectively. On the other hand, a quantitative analysis of TA and virulence genes revealed that the expression level in both genes is different under normal and stress conditions. The results obtained by anti-mazF peptide nucleic acids demonstrated that the expression level of virulence genes had decreased. These findings demonstrate an association between TA systems and virulence factors. The mazEF on the plasmids and the higBA TA genes on the chromosomes of all E. faecium and E. faecalis strains were dominant. Additionally, there was a decrease in the expression of virulence genes in the presence of anti-mazF peptide nucleic acids. Therefore, it is suggested that mazEF TA systems are potent and sensitive targets in all E. faecium and E. faecalis strains.
Soheili, Sara; Ghafourian, Sobhan; Sekawi, Zamberi; Neela, Vasantha Kumari; Sadeghifard, Nourkhoda; Taherikalani, Morovat; Khosravi, Afra; Ramli, Ramliza; Hamat, Rukman Awang
The toxin–antitoxin (TA) system is a regulatory system where two sets of genes encode the toxin and its corresponding antitoxin. In this study, the prevalence of TA systems in independently isolated clinical isolates of Enterococcus faecium and Enterococcus faecalis was determined, the dominant TA system was identified, different virulence genes in E. faecium and E. faecalis were surveyed, the level of expression of the virulence and TA genes in normal and stress conditions was determined, and finally their associations with the TA genes were defined. Remarkably, the analysis demonstrated higBA and mazEF in all clinical isolates, and their locations were on chromosomes and plasmids, respectively. On the other hand, a quantitative analysis of TA and virulence genes revealed that the expression level in both genes is different under normal and stress conditions. The results obtained by anti-mazF peptide nucleic acids demonstrated that the expression level of virulence genes had decreased. These findings demonstrate an association between TA systems and virulence factors. The mazEF on the plasmids and the higBA TA genes on the chromosomes of all E. faecium and E. faecalis strains were dominant. Additionally, there was a decrease in the expression of virulence genes in the presence of anti-mazF peptide nucleic acids. Therefore, it is suggested that mazEF TA systems are potent and sensitive targets in all E. faecium and E. faecalis strains. PMID:26005332
Borzini, Letizia; Condò, Roberta; De Dominicis, Paolo; Casaglia, Adriano; Cerroni, Loredana
Background: There are various microorganisms related to intra and extra-radicular infections and many of these are involved in persistent infections. Bacterial elimination from the root canal is achieved by means of the mechanical action of instruments and irrigation as well as the antibacterial effects of the irrigating solutions. Enterococcus faecalis can frequently be isolated from root canals in cases of failed root canal treatments. Antimicrobial agents have often been developed and optimized for their activity against endodontic bacteria. An ideal root canal irrigant should be biocompatible, because of its close contact with the periodontal tissues during endodontic treatment. Sodium hypoclorite (NaOCl) is one of the most widely recommended and used endodontic irrigants but it is highly toxic to periapical tissues. Objectives: To analyze the literature on the chemotherapeutic agent and plant extracts studied as root canal irrigants. In particularly, the study is focused on their effect on Enterococcus faecalis. Method: Literature search was performed electronically in PubMed (PubMed Central, MEDLINE) for articles published in English from 1982 to April 2015. The searched keywords were “endodontic irrigants” and “Enterococcus faecalis” and “essential oil” and “plant extracts”. Results: Many of the studied chemotherapeutic agents and plant extracts have shown promising results in vitro. Conclusion: Some of the considered phytotherapic substances, could be a potential alternative to NaOCl for the biomechanical treatment of the endodontic space. PMID:28217184
Full Text Available Abstract The aim was to evaluate the functional efficiency of a probiotic strain Enterococcus faecium M74 in the feed on egg yolk weight, egg yolk fat and cholesterol contents of Shaver Starcross 288 hens. Feed in the experimental group was enriched with a probiotic additive containing of 5*109 viable Enterococcus faecium per g. Egg samples a total 30 pcs per group were collected during the first egg-laying period at week 28 and 38 of hens´ age. Non-significantly lower of egg yolk weight was observed in the experimental group at all sampling times compared with their respective controls. Non-significantly lower of egg yolk weight was observed in the experimental group at all sampling times compared with their respective controls. Significantly lower concentrations of egg yolk cholesterol were found in the experimental group at week 28, and week 38 in compare with controls. In conclusion, the addition of probiotic strains Enterococcus faecium M74 to the feed of Shaver Starcross 288 hens reduced cholesterol in egg yolk at all sampling times. Even though the hypocholesterolemic mechanism of probiotics has not yet been fully understood, it is an established fact that cholesterol and bile salt metabolism are closely linked. However, the hypocholesterolemic mechanism of probiotics based on the bile salt hydrolase activity hypothesis has not yet been sufficiently elucidated.
I Wayan Suardana
Full Text Available Lactic acid bacteria can excrete antimicrobial compounds like bacteriocins. The study aimed to find out the characteristic of physic-chemical of bacteriocin producing Enterococcus durans isolate 18A isolated from colon’s bali cattle. The study initiated by Gram staining and catalase test, followed by isolation and purification of bacteriocin. The result of the research showed that bacteriocins of Enterococcus durans isolate 18A as a protein with it’s concentrationis 0,272?g/ml and it does not contain carbohydrate. On the other hand, the bacteriocins was not showed a band while tested on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE. The result showed, the bacteriocin producing Enterococcus durans isolate 18A has antimicrobial activity to Bacillus cereus as 23,88%.
Marcos J. Carruitero
Full Text Available Objective: To compare the in vitro antibacterial effect of the root canal cements Endobalsam®, Top Seal®, Apexit® and Endofill® against Enterococcus faecalis ATCC 29212. Materials and method: Eighty-five applications of cements on Enterococcus faecalis, cultured in vitro on solid media in Petri dishes, were analyzed. Five groups were evaluated: four for each cement, and the fifth for the positive control (amoxicillin. The antibacterial effect was measured by the diameters of the bacterial inhibition halos at 24 hours, 48 hours, and seven days. Student´s t-test, ANOVA and the Tukey test were used for the statistical analysis. Results: No statistically significant differences were found at 24 hours (p>0.05; at 48 hours and seven days, Endofill® and Apexit® had the greatest effect (p0.05. Conclusion: Enterococcus faecalis ATCC 29212 was susceptible to all cements. Endofill® had greater in vitro antibacterial effect than Apexit®, Top Seal® and Endobalsam®.
Tu Hoang Khue Nguyen
Full Text Available Puffer fishes were collected from the central sea in Vietnam from spring to summer season. The eggs were incubated in MRS broth that was used to test the toxicity in mice and isolate the lactic acid bacteria community that could produce tetrodotoxin (TTX. Thin layer chromatography (TLC and high performance lipid chromatography (HPLC were used to detect and quantify TTX. As a result, Enterococcus faecium AD1 which was identified by biochemical test and 16S rRNA analysis could produce TTX 0.3 mg/mL when cultured in MRS broth. The bacterium was optimized for TTX production and gave 0.18 mg/mL, 0.07 mg/mL, and 0.15 mg/mL in media prepared from the meat-washing water of freshwater fishes (Pangasius bocourti, Oreochromis sp. and sea fish (Auxis thazard, respectively, that are also hopeful to answer some poisoning cases related to eating fishes. Enterococcus faecium also showed the wide antimicrobial activities on yeast, Gram-negative and -positive bacteria. Extracted exopolysaccharide (EPS that reacted with 2,2-diphenyl-1-picrylhydrazyl to give IC50 at 5 mg/mL equaled 11 mg/mL ascorbic acid which could show effects on Hela-6 and Hep G2 using sulforhodamine B test. Enterococcus faecium can be claimed as a promising source in tetrodotoxin and biological compounds.
Vanessa Stolf Boretti
Full Text Available Objetivo: Observar a presença de microrganismos dos gêneros Staphylococcus e Streptococcus em brinquedos de uma brinquedoteca de unidade pediátrica hospitalar, bem como analisar o perfil de resistência aos antimicrobianos das cepas isoladas. Métodos: Foram realizadas coletas de 60 brinquedos, utilizando swab umedecido em solução fisiológica, logo após a utilização pelas crianças. As amostras coletadas foram semeadas em meios de cultura para proporcionar o isolamento dos microrganismos e posterior identificação. Foi realizado o antibiograma para todas as bactérias identificadas, com a técnica de difusão em agar. Resultados: O gênero Staphylococcus estava presente em 87% (52/60 dos brinquedos analisados. Foram isoladas 73 cepas, sendo 29% (21/73 de Staphylococcus coagulasepositiva e 71% (52/73 de Staphylococcus coagulase-negativa. Neste estudo, 90,4% das cepas coagulase-negativas apresentaram resistência à penicilina, 65,4% à oxacilina, 28,8% à claritromicina, 61,5% à clindamicina e nenhuma à vancomicina. Das cepas coagulase-positivas, 76,2% apresentaram resistência à penicilina, 23,8% à oxacilina, 23,8% à claritromicina, 47,6% à clindamicina e nenhuma à vancomicina. Não foram detectadas bactérias do gênero Streptoccocus nos brinquedos estudados. Conclusões: Os resultados obtidos demonstraram que os brinquedos podem apresentar contaminação por bactérias potencialmente patogênicas com resistência aos antimicrobianos, representando uma possível fonte de infecção nosocomial para pacientes que normalmente já se encontram debilitados.
Claudia M. Romero-Vivas
Full Text Available La leptospirosis, la enfermedad bacteriana zoonótica y emergente más importante en el mundo, es causada por las especies patógenas de Leptospira spp. Han sido descritas veinte especies de Leptospira spp.; se ha determinado la secuencia del ADN genómico de algunas cepas patógenas; la función de la mayoría de los genes involucrados en su patogénesis permanece desconocida. La leptospirosis humana presenta un rango de síntomas que van desde una fiebre indiferenciada hasta una ictericia, hemorragia, fallas renales y pulmonares severas. La administración temprana e intravenosa de penicilina G es requerida para reducir las tasas de mortalidad, pero los antibióticos pueden no ser efectivos en la enfermedad pulmonar severa. En las Américas, las áreas de alto riesgo son Brasil, Centroamérica y el Caribe. En Colombia se han realizado pocos estudios. La prueba serológica de oro, la microaglutinación, tiene alta sesibilidad y especificidad cuando se usan baterías de serovariedades locales, pero es serogrupo específica. Las vacunas generan respuestas específicas para la serovariedad usada, pero no previenen la infección o trasmisión. Problemas en el diagnóstico de laboratorio de la leptospirosis conllevan a un subregistro en el número de casos; altas tasas de mortalidad asociadas a fallas renal y pulmonar son resultado de las dificultades en el manejo de los casos.
Aragonés, L; López, I; Palazón, A; López-Úbeda, R; García, C
Sun. and beach tourism is very important to the economy of Spain, so the control of the quality of the environment on the beaches is essential. Therefore, the analysis and control of the quality of bathing water is necessary, which is defined by the European Directive 2006/7/EC as excellent, good or sufficient depending on the presence of microbiological contamination or other organisms or waste presenting a risk to bathers' health. For that, 1392 beaches of the Iberian Peninsula and its islands were analysed, taking into account: fecal bacteria (Escherichia coli and Enterococcus), physical characteristics of sediment, level of urbanization, climatic and anthropogenic factors, and maritime climate. Thus, it was observed that urban sand beaches located in seas with fewer hours of sunshine and important tide have higher concentrations of E. coli and Enterococcus. There is also an indirect relationship between these microorganisms with salinity (R(2) 0.746 for E. coli and 0.606 for Enterococcus), temperature (R(2) 0.743 for E. coli and 0.604 for Enterococcus) and hours of sunshine (R(2) 0.781 for E. coli and 0.706 for Enterococcus), while this relationship is direct with rainfall (R(2) 0.640 for E. coli and 0.607 for Enterococcus) or wave height (R(2) 0.769 for E. coli and 0.601 for Enterococcus). From all this, it follows that the Directive 2006/7/EC should define more specific criteria as to the place and time of sampling, and take into account the different environment variables that influence the survival of bacteria, so that the results may reflect reality, and avoid staff responsible for sampling freely choose the place and time of sampling. Copyright © 2016 Elsevier B.V. All rights reserved.
Patel, Khilna; Kabir, Rubiya; Ahmad, Samrah; Allen, Steven L
The incidence and severity of vancomycin-resistant Enterococcus blood stream infections continue to rise and is a significant burden in the healthcare setting. Literature thus far is minimal regarding treatment outcomes in patients with malignancy and vancomycin-resistant Enterococcus bacteremia. Appropriate antibiotic selection is vital to treatment success due to high rates of resistance, limited antimicrobials and mortality in this patient population. We conducted this study to determine whether treatment outcomes differed between cancer patients treated with linezolid and those treated with daptomycin for vancomycin-resistant Enterococcus bacteremia. This single-center, retrospective study included adult patients hospitalized on the oncology service with documented vancomycin-resistant Enterococcus faecium or Enterococcus faecalis bacteremia who received at least 48 h of either linezolid or daptomycin as primary treatment. A total of 65 patients were included in the analysis. Thirty-two patients received daptomycin as primary treatment, and 33 patients received linezolid as primary treatment. Twenty-six (76.5%) patients in the linezolid cohort versus 22 (71%) patients in the daptomycin cohort achieved microbiological cure (p = 0.6141). Median length of stay in days (30 vs. 42, p = 0.0714) and mortality (7/32 (20.6%) vs. 8/33 (25.8%), p = 0.6180) were also similar between the linezolid and daptomycin treated patients, respectively. No differences in microbiological cure, length of stay or mortality were identified between the groups. This study suggests that linezolid and daptomycin are each reasonable options for treating vancomycin-resistant Enterococcus bacteremia in oncology patients. Further prospective, randomized controlled trials are needed to assess the optimal treatment for vancomycin-resistant Enterococcus bacteremia in this patient population. © The Author(s) 2014.
Graciana Maria de Moraes
Full Text Available OBJETIVO: Identificar os fatores preditores de infecção ou colonização por micro-organismos resistentes. MÉTODOS: Foi realizado estudo quantitativo de coorte prospectivo. Foram realizadas a análise descritiva, para conhecimento da população do estudo, e a análise discriminante, para identificação dos fatores preditores. RESULTADOS: Foram incluídos 85 pacientes com infecções por micro-organismos resistentes: Pseudomonas aeruginosas resistente aos carbapenêmicos (24,7%, Acinetobacter resistente aos carbapenêmicos (21,2%, Staphylococcus aureus resistente à meticilina (25,9%, Enterococcus spp. resistente à vancomicina (17,6% e Klebsiella pneumoniae resistente aos carbapenêmicos (10,6%. A análise discriminante identificou transferências de outros hospitais e internação na Unidade de Terapia Intensiva como fatores preditores para ocorrência de infecção pelos grupos S. aureus resistente à meticilina, Acinetobacter resistente aos carbapenêmicos e K. pneumoniae resistente aos carbapenêmicos. Nenhuma das variáveis estudadas foi discriminante para Enterococcus spp. resistente à vancomicina e P. aeruginosas resistente aos carbapenêmico. CONCLUSÃO: Os fatores preditores encontrados foram: internação na UTI e a transferências de outros hospitais.OBJECTIVE: Identifying predictors of infection or colonization with resistant microorganisms. METHODS: A quantitative study of prospective cohort was carried out. A descriptive analysis was performed in order to know the population of the study and a discriminant analysis was performed to identify the predictors. RESULTS: In this study were included 85 patients with infections caused by resistant microorganisms: carbapenem-resistant Pseudomonas aeruginosas (24.7%; carbapenem-resistant Acinetobacter (21.2%; methicillin-resistant Staphylococcus aureus (25.9%, vancomycin-resistant Enterococcus spp (17.6% and carbapenem-resistant Klebsiella pneumonia (10.6%. The discriminant analysis
Antimicrobial susceptibility among Enterococcus isolates from the city of Porto Alegre, RS, Brazil Sensibilidade aos antimicrobianos entre amostras de Enterococcus isolados na cidade de Porto Alegre, RS, Brasil
Pedro Alves d'Azevedo
. faecalis (92,8%, seguidas por E. faecium (2,9%, E. gallinarum (1,5%, E. avium (1,1%, E. hirae (0,7%, E. casseliflavus (0,4%, E. durans (0,4% e E. raffinosus (0,2%. Os testes de sensibilidade revelaram que 62,0% das amostras foram resistentes à tetraciclina, 42,6% à eritromicina, 24,8% ao cloranfenicol, 22,6% à ciprofloxacina, 22,0% à norfloxacina, 3,5% à ampicilina e 3,5% á nitrofurantoína. Resistência a níveis elevados de aminoglicosídeos foi encontrada em 37,8% das amostras, com 23,5% sendo resistente à gentamicina, 14,3% à estreptomicina e 2,8% a ambos gentamicina e estreptomicina. Nenhuma amostra com resistência adquirida à vancomicina ou produtora de b-lactamase foi encontrada. Os resultados indicam um significante percentual de amostras resistentes a diferentes antimicrobianos, apontando a necessidade de estratégias de controle para evitar a disseminação de cepas resistentes e de vigilância contínua para a detecção de características emergentes de resistência.
Starting next year gas utility Slovensky plynarensky priemysel, a.s., Bratislava (SPP) will decrease the fees for transport of gas to its client from third parties. This decision should have a positive effect mainly on large industrial customers. The fact that SPP decided not to apply the approved ruling to its full extent was the result of negotiations with the regulator and the Ministry of Economy. SPP is not afraid it might lose customers. This decision only gives them a competitive advantage. (Author)
Fouad, A F; Kum, K-Y; Clawson, M L; Barry, J; Abenoja, C; Zhu, Q; Caimano, M; Radolf, J D
Eubacterium spp. and Streptococcus spp. are virulent, commonly identified microorganisms in endodontic infections. The purpose of this study was to use molecular methods to identify these organisms in 22 infected root canals that include eight cases with preoperative clinical symptoms and five cases with a history of diabetes mellitus. The presence of Streptococcus spp. and Eubacterium spp. was examined using two sets of PCR primers specific with multiple species within the respective genera. Positive specimens had their PCR products sequenced and phylogenetically analyzed to identify the specific species. Sixteen specimens (73%) contained Eubacterium spp. and nine (41%) were positive for Streptococcus spp. Eubacterium infirmum was the most prevalent Eubacterium sp. This organism was significantly associated with a history of diabetes (OR = 9.6; P = 0.04). Streptococcus anginosus was the most common Streptococcus sp., but neither it nor any of the other streptococci were significantly associated with the clinical parameters evaluated.
Freitas, Ana R.; Coque, Teresa M.; Novais, Carla
clonally related Enterococcus faecium clonal complex 5 (CC5) isolates (17 sequence type 6 [ST6], 6 ST5, 5 ST185, 1 ST147, and 1 ST493) were obtained from feces of swine and healthy humans. This collection included isolates widespread among pigs of European Union (EU) countries since the mid-1990s. Each ST...... comprised isolates showing similar pulsed-field gel electrophoresis (PFGE) patterns (≤6 bands difference; >82% similarity). Some CC5 PFGE subtype strains from swine were indistinguishable from hospital vancomycin-resistant enterococci (VRE) causing infections. A truncated variant of Tn1546 (encoding...... resistance to vancomycin) and tcrB (coding for resistance to copper) were consistently located on 150- to 190-kb plasmids (rep(pLG1)). E. faecium CC17 (ST132) isolates from pig manure and two clinical samples showed identical PFGE profiles and contained a 60-kb mosaic plasmid (rep(Inc18) plus rep...
Petra Regine Adelheid Kohler
Full Text Available The discovery of the third domain of life, the Archaea, is one of the most exciting findings of the last century. These remarkable prokaryotes are well known for their adaptations to extreme environments; however, Archaea have also conquered moderate environments. Many of the archaeal biochemical processes, such as methane production, are unique in nature and therefore of great scientific interest. Although formerly restricted to biochemical and physiological studies, sophisticated systems for genetic manipulation have been developed during the last two decades for methanogenic archaea, halophilic archaea and thermophilic, sulfur-metabolizing archaea. The availability of these tools has allowed for more complete studies of archaeal physiology and metabolism and most importantly provides the basis for the investigation of gene expression, regulation and function. In this review we provide an overview of methods for genetic manipulation of Methanosarcina spp., a group of methanogenic archaea that are key players in the global carbon cycle and which can be found in a variety of anaerobic environments.
Khalkhali, Soodabeh; Mojgani, Naheed
Background and Objectives: Human milk is a continuous supply of Lactic Acid bacteria (LAB), including enterococci with probiotic potentials. The aim of this study was to analyze two Enterococcus species, isolated from human milk for their probiotic potential, bacteriocin producing ability and virulence traits. Materials and Methods: Enterococcus faecium TA0033 and E. faecalis TA102 were tested for acid and bile tolerance, survival in simulated gastric and intestinal conditions. The antibacterial spectrum of the isolates was tested by agar well diffusion assay. The antagonistic agent was characterized by physico-chemical methods. The enterocin structural genes, virulence determinants, vancomycin resistance and biogenic amine genes, such as hdc1, hdc2, tdc, ldc and odc were also determined. Results: The tested isolates survived acidic conditions, high bile salt (1%), simulated gastric and intestinal conditions. The culture supernatant fluids of the two isolates inhibited the growth of Escherichia coli, Listeria monocytogenes, Salmonella typhi, Staphylococcus aureus, Shigella dysenteriae and Streptococcus agalactiae. The antagonistic activity was lost in the presence of proteolytic enzymes but tolerated the action of catalase, lysozyme and lipase. In contrast to enterocin TA102, enterocin TA0033 possessed bactericidal mode of action. Bacteriocin structural genes, entA and entB were present in the genome of the two isolates, while E. faecalis TA102 additionally harboured entP and bac31 genes. The phenotypic and genotypic virulence assessment studies indicated hyaluronidase (hyl) production and vancomycin resistance in E. faecalis TA102 while, none of the isolates harboured the biogenic amine genes. Conclusion: The presence of virulence genes in E. faecalis TA102 calls for careful monitoring of Enterococcus isolates for their safety parameters. PMID:29238458
Herranz, C.; Chen, Y.; Chung, H.-J.; Cintas, L. M.; Hernández, P. E.; Montville, T. J.; Chikindas, M. L.
Enterocin P is a pediocin-like, broad-spectrum bacteriocin which displays a strong inhibitory activity against Listeria monocytogenes. The bacteriocin was purified from the culture supernatant of Enterococcus faecium P13, and its molecular mechanism of action against the sensitive strain E. faecium T136 was evaluated. Although enterocin P caused significant reduction of the membrane potential (ΔΨ) and the intracellular ATP pool of the indicator organism, the pH gradient (ΔpH) component of the proton motive force (Δp) was not dissipated. By contrast, enterocin P caused carboxyfluorescein efflux from E. faecium T136-derived liposomes. PMID:11282622
Herranz, Carmen; Cintas, Luis M.; Hernández, Pablo E.; Moll, Gert N.; Driessen, Arnold J. M.
Enterocin P is a bacteriocin produced by Enterococcus faecium P13. We studied the mechanism of its bactericidal action using enterocin-P-sensitive E. faecium T136 cells. The bacteriocin is incapable of dissipating the transmembrane pH gradient. On the other hand, depending on the buffer used, enterocin P dissipates the transmembrane potential. Enterocin P efficiently elicits efflux of potassium ions, but not of intracellularly accumulated anions like phosphate and glutamate. Taken together, these data demonstrate that enterocin P forms specific, potassium ion-conducting pores in the cytoplasmic membrane of target cells. PMID:11181377
Hamidi, Moufida; Ammari, Houria; Ghaffor, Mohamed; Benamrouche, Nabila; Tali-Maamar, Hassiba; Tala-Khir, Farida; Younsi, Mokhtar; Rahal, Kheira
A glycopeptide-resistant Enterococcus faecium (EFRG) was isolated from a wound in a patient hospitalized in a university hospital in Algiers. This strain was resistant to several antibiotics. This patient was carrying this strain in the digestive tract which may partly explain its origin. Genotypic comparison of the two strains by pulsed field gel electrophoresis showed that it was the same strain. Glycopeptide resistance was due to the presence of the vanA gene. Vigilance is required facing the emergence of strains of EFRG in our hospitals.
Thompson, Mickala M; Hassoun, Ali
Infective endocarditis (IE) one-year mortality rates approach 40%. Here, we report two native valve Enterococcus faecalis IE cases in patients successfully treated with telavancin. An 88-year-old with mitral valve endocarditis and a penicillin allergy, initially treated with intravenous vancomycin, was switched to telavancin. A 69-year-old, who previously received amoxicillin and intravenous vancomycin for presumed enterococcal bacteraemia, was diagnosed with dual valve endocarditis for which he received telavancin. Both received six weeks of telavancin. Neither had telavancin-related adverse events, evidence of infection at six months, nor required telavancin dosing adjustments. Documented use of novel treatments for serious enterococcal infections is needed.
Pashkova, Tatiana M; Vasilchenko, Alexey S; Khlopko, Yuriy A; Kochkina, Elena E; Kartashova, Olga L; Sycheva, Maria V
We report here the complete genome sequence of Enterococcus faecium strain ICIS 96, which was isolated from the feces of a horse. Bacteriological characterization of strain ICIS 96 revealed the absence of pathogenicity factors, while its spectrum of antagonistic activity was found to be broad, having activities associated with both Gram-positive and Gram-negative bacteria. Analysis of the E. faecium ICIS 96 genome revealed five genes associated with antimicrobial activity (enterocin [ent] A, ent B, lactobin A/cerein 7b, and ent L50 A/B). No genes that correlate with human pathogenicity were identified. Copyright © 2018 Pashkova et al.
Brachkova, Mariya I; Duarte, Maria A; Pinto, João F
The objective of the study was to produce calcium alginate beads able to deliver Lactobacillus spp. (Lactobacillus plantarum, Lactobacillus rhamnosus GG, Lactobacillus bulgaricus and Lactobacillus lactis) with preserved viability and antibacterial activity. Four types of beads, containing entrapped (E), surface and entrapped (ES), surface (S) and concentrated surface and entrapped lactobacilli (C(ES)) were prepared and physically characterized. The antibacterial activity of lactobacilli cultures before and after immobilization, freeze-drying and throughout storage was studied in relationship to the viable number of lactobacilli. Multi-resistant clinical isolates (methicillin-resistant Staphylococcus aureus, vancomycine-resistant Enterococcus faecalis, VIM-2-metalo-β-lactamase producing Pseudomonas aeruginosa and CTX-M-15-β-lactamase producing strains: Escherichia coli and Klebsiella pneumoniae) were used as indicator strains. Alginate beads in which lactobacilli proliferated to the beads surface (ES and C(ES)) differed significantly from the other types of beads in their physicochemical properties, showing smoother surface morphology, more spherical shape, bigger weight, lower calcium content, density and crushing force. Lactobacilli cultures, at high cell concentrations (10(8)cfu/ml) were active against both Gram-positive and negative multi-resistant bacteria. Beads containing both entrapped and surface lactobacilli (ES) resulted in viability and antibacterial activity most similar to non-processed lactobacilli cultures. The viability and antibacterial activity of the immobilized lactobacilli remained stable after 6 months storage. Copyright © 2010 Elsevier B.V. All rights reserved.
Full Text Available Abstract Background Only limited information is available about the occurrence of ticks and tick-borne pathogens in public parks, which are areas strongly influenced by human beings. For this reason, Ixodes ricinus were collected in public parks of different Bavarian cities in a 2-year survey (2009 and 2010 and screened for DNA of Babesia spp., Rickettsia spp. and Bartonella spp. by PCR. Species identification was performed by sequence analysis and alignment with existing sequences in GenBank. Additionally, coinfections with Anaplasma phagocytophilum were investigated. Results The following prevalences were detected: Babesia spp.: 0.4% (n = 17, including one pool of two larvae in 2009 and 0.5 to 0.7% (n = 11, including one pool of five larvae in 2010; Rickettsia spp.: 6.4 to 7.7% (n = 285, including 16 pools of 76 larvae in 2009. DNA of Bartonella spp. in I. ricinus in Bavarian public parks could not be identified. Sequence analysis revealed the following species: Babesia sp. EU1 (n = 25, B. divergens (n = 1, B. divergens/capreoli (n = 1, B. gibsoni-like (n = 1, R. helvetica (n = 272, R. monacensis IrR/Munich (n = 12 and unspecified R. monacensis (n = 1. The majority of coinfections were R. helvetica with A. phagocytophilum (n = 27, but coinfections between Babesia spp. and A. phagocytophilum, or Babesia spp. and R. helvetica were also detected. Conclusions I. ricinus ticks in urban areas of Germany harbor several tick-borne pathogens and coinfections were also observed. Public parks are of particularly great interest regarding the epidemiology of tick-borne pathogens, because of differences in both the prevalence of pathogens in ticks as well as a varying species arrangement when compared to woodland areas. The record of DNA of a Babesia gibsoni-like pathogen detected in I. ricinus suggests that I. ricinus may harbor and transmit more Babesia spp. than previously known. Because of their high recreational value for human beings, urban green
Schorn, Sabine; Pfister, Kurt; Reulen, Holger; Mahling, Monia; Silaghi, Cornelia
Only limited information is available about the occurrence of ticks and tick-borne pathogens in public parks, which are areas strongly influenced by human beings. For this reason, Ixodes ricinus were collected in public parks of different Bavarian cities in a 2-year survey (2009 and 2010) and screened for DNA of Babesia spp., Rickettsia spp. and Bartonella spp. by PCR. Species identification was performed by sequence analysis and alignment with existing sequences in GenBank. Additionally, coinfections with Anaplasma phagocytophilum were investigated. The following prevalences were detected: Babesia spp.: 0.4% (n = 17, including one pool of two larvae) in 2009 and 0.5 to 0.7% (n = 11, including one pool of five larvae) in 2010; Rickettsia spp.: 6.4 to 7.7% (n = 285, including 16 pools of 76 larvae) in 2009. DNA of Bartonella spp. in I. ricinus in Bavarian public parks could not be identified. Sequence analysis revealed the following species: Babesia sp. EU1 (n = 25), B. divergens (n = 1), B. divergens/capreoli (n = 1), B. gibsoni-like (n = 1), R. helvetica (n = 272), R. monacensis IrR/Munich (n = 12) and unspecified R. monacensis (n = 1). The majority of coinfections were R. helvetica with A. phagocytophilum (n = 27), but coinfections between Babesia spp. and A. phagocytophilum, or Babesia spp. and R. helvetica were also detected. I. ricinus ticks in urban areas of Germany harbor several tick-borne pathogens and coinfections were also observed. Public parks are of particularly great interest regarding the epidemiology of tick-borne pathogens, because of differences in both the prevalence of pathogens in ticks as well as a varying species arrangement when compared to woodland areas. The record of DNA of a Babesia gibsoni-like pathogen detected in I. ricinus suggests that I. ricinus may harbor and transmit more Babesia spp. than previously known. Because of their high recreational value for human beings, urban green areas are likely to remain in the research focus on
Realpe-Delgado, María Elena
Full Text Available Salmonella spp., Campylobacter spp., and L. monocytogenes are zoonotic foodborne pathogens, associated with the consumption of contaminated foods of animal origin. In this study we determined the prevalence and risk factors associated with the presence of these microorganisms at all stages of the production system, in two Colombian poultry companies (EI-EI-I and II. In EI-I, Campylobacter spp., and Salmonella spp., were isolated from 10 % and 4.4 % of the specimens, and S. Heidelberg was the predominant serotype. Salmonella spp., was found in 6 % of hands and stool samples of workers. S. Saphra was the most prevalent serotype. In EI-II, the prevalence of Campylobacter spp., and Salmonella spp., from animal specimens was 7 % and 17 %, respectively. L. monocytogenes was not detected. This study established the prevalence of these zoonotic pathogens through the production chain and showed the presence of pathogen carriers among workers/food handlers. “Lack of medical examination of employees in the previous year” was found to be a possible risk factor for carriage of Salmonella spp.
Furustrand Tafin, U.; Meis, J.F.G.M.; Trampuz, A.
We evaluated isothermal microcalorimetry for real-time susceptibility testing of non-Aspergillus molds. MIC and minimal effective concentration (MEC) values of Mucorales (n = 4), Fusarium spp. (n = 4), and Scedosporium spp. (n = 4) were determined by microbroth dilution according to the Clinical
Bermudagrass (Cynodon spp.) and zoysiagrass (Zoysia spp.), which are both used as warm-season turfgrasses in the United States, are members of subfamily Chloridoideae and are reported to be at least 55% genetically similar. To assess if molecular tools between the two species can be interchanged, 93...
Falkinham, Joseph O; Williams, Myra D; Kwait, Rebecca; Lande, Leah
A published survey of bacteria in showerhead biofilm samples revealed that Methylobacterium spp. and Mycobacterium spp. seldom coexisted in biofilms. To confirm that information, biofilm samples were collected from household plumbing of Mycobacterium avium patients and Methylobacterium spp. and M. avium numbers were measured by direct colony counts. The results demonstrated that if Methylobacterium spp. were present, Mycobacterium spp. were absent, and the opposite. The data demonstrate that microbial populations in biofilms can influence the presence or absence of opportunistic premise plumbing pathogens and, thereby, increase the range of strategies to reduce exposure to waterborne pathogens. Finally, by assessing for the visual presence of methylobacteria as pink pigmentation on showers and shower curtains, homeowners and managers of hospitals and other buildings can quickly determine whether a premise plumbing biofilm sample has mycobacteria with a high degree of assurance. Copyright © 2016 Asian African Society for Mycobacteriology. Published by Elsevier Ltd. All rights reserved.
Ogihara, Shinji; Saito, Ryoichi; Sawabe, Etsuko; Hagihara, Michio; Tohda, Shuji
A male patient was admitted to our hospital due to infectious endocarditis. He had been treated with levofloxacin for 6 weeks, sulbactam/cefoperazone for 4 weeks, and benzylpenicillin for 2 days prior to valve replacement surgery. Gram-positive cocci, with morphology consistent with γ-Streptococcus, were detected in blood cultures obtained at admission, as well as in vegetation obtained from the aortic valve. However, the strain could not be identified using biochemical methods. Sequencing of the 16S rRNA gene indicated that the culture was a small-colony variant of Enterococcus faecalis. This is the first case in Japan of infectious endocarditis due to E. faecalis small-colony variants. Small-colony variants are subpopulations of bacteria with slow growth, reduced sugar fermentation, and unstable phenotype. As a result, these strains tend to be misidentified. Further, small-colony variants are associated with recurrent and persistent infections such as prosthetic joint infection and infectious endocarditis. These strains are found in various bacterial species, including Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas aeruginosa, but rarely in Enterococcus species. The case highlights the need to be vigilant of E. faecalis small-colony variants, especially in patients who received long-term courses of antibiotics. Copyright © 2016 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.
Chung, Yeon Soo; Kwon, Ka Hee; Shin, Sook; Kim, Jae Hong; Park, Yong Ho; Yoon, Jang Won
Possible cross-transmission of hospital-associated enterococci between human patients, medical staff, and hospital environments has been extensively studied. However, limited information is available for veterinary hospital-associated Enterococcus isolates. This study investigated the possibility of cross-transmission of antibiotic-resistant enterococci between dog patients, their owners, veterinary staff, and hospital environments. Swab samples (n =46 5) were obtained from five veterinary hospitals in Seoul, Korea, during 2011. Forty-three Enterococcus strains were isolated, representing seven enterococcal species. E. faecalis and E. faecium were the most dominant species (16 isolates each, 37.2%). Although slight differences in the antibiotic resistance profiles were observed between the phenotypic and the genotypic data, our antibiogram analysis demonstrated high prevalence of the multiple drug-resistant (MDR) isolates of E. faecalis (10/16 isolates, 62.5%) and E. faecium (12/16 isolates, 75.0%). Pulsed-field gel electrophoretic comparison of the MDR isolates revealed three different clonal sets of E. faecalis and a single set of E. faecium, which were isolated from different sample groups or dog patients at the same or two separate veterinary hospitals. These results imply a strong possibility of cross-transmission of the antibiotic-resistant enterococcal species between animal patients, owners, veterinary staff, and hospital environments.
Bager, Flemming; Aarestrup, Frank Møller; Madsen, Mogens
The use of the glycopeptide growth promoter avoparcin was discontinued in Denmark in 1995 following concerns that vancomycin-resistant Enterococcus faecium occurring as a result of its use could be transferred to humans via food. The present study is an analysis of results obtained by the continu......The use of the glycopeptide growth promoter avoparcin was discontinued in Denmark in 1995 following concerns that vancomycin-resistant Enterococcus faecium occurring as a result of its use could be transferred to humans via food. The present study is an analysis of results obtained...... by the continuous surveillance of an antimicrobial resistance in Denmark (DANMAP) with the aim of determining the effect of the ban on the occurrence of glycopeptide resistance among E, faecium isolated from broilers and pigs, Among isolates from broilers, the proportion that were resistant to glycopeptides has....... Alternatively, the results indicate that the different outcomes may result from different co-selection patterns in pigs and broilers. In pigs, the antimicrobials most commonly used favored co-selection of glycopeptide-resistant strains of E. faecium while in broilers the antimicrobials most widely used selected...
Mohammad Esmaeel Ebrahimi Chaharom
Full Text Available Background and aims. The aim of this study was to evaluate the antibacterial activity of one-step self-etch adhesives on Enterococcus faecalis on days 1, 7 and 14 with the use of modified direct contact test. Materials and methods. The modified directcontact test was used to evaluate the antibacterial effect of Adper Easy One, Bond Force, Clearfil S3 Bond, Futurabond M, G-Bond, iBond and OptiBond All-in-one adhesives on Enterococcus faecalisafter aging the samples in phosphate-buffered saline for one, seven and fourteen days. Data were analyzed using one-way ANOVA and post hoc Tukey tests. Aging effect of each adhesive was evaluated by paired-sample test. In this study, P0.05. Conclusion. iBond exhibited the highest antibacterial effect on E. faecalis after one week. Futurabond and OptiBond All-in-one exhibited antibacterial effects against E. faecalis for one week.
Julio C. Hernandez-Camarena
Full Text Available Background: Vancomycin-resistant Enterococcus (VRE endophthalmitis after penetrating keratoplasty (PKP is very rare, the management is a challenge due to both the pattern of antibiotic resistance and the aggressive nature of the infectious process. We report the first delayed-onset case of VRE endophthalmitis after PKP. Materials and Methods: Case report of a 51-year-old female with a 7-week history of PKP who arrived at the emergency room with signs and symptoms of endophthalmitis. Initial visual acuity was light perception, and a posterior pole exam was not possible due to the intense vitreous reaction. Mode B ultrasound was used to assess the posterior pole. The patient underwent pars plana vitrectomy and received intravitreous antibiotics. Results: Vitreous stains and cultures were positive for Enterococcus faecium resistant to vancomycin. Donor rim cultures and viral PCR were negative. Treatment was carried out by repeated intravitreal antibiotics and systemic linezolid. Clinical improvement was seen after the second dose of intravitreous antibiotics and systemic linezolid, but visual acuity remained at light perception consistent with the ischemic changes observed in the posterior pole. Conclusion: VRE endophthalmitis might be associated with positive donor rim cultures. Prompt use of systemic linezolid in addition to intravitreous antibiotics is recommendable, but even with prompt treatment, visual prognosis is guarded.
Diane S. Daniel
Full Text Available Enterococcus faecalis ranks as one of the leading causes of nosocomial infections. A strong epidemiological link has been reported between E. faecalis inhabiting animals and environmental sources. This study investigates the genetic diversity, antibiotic resistance and virulence determinants in E. faecalis from three sources in Malaysia. A total of 250 E. faecalis isolates were obtained consisting of 120 isolates from farm animals, 100 isolates from water sources and 30 isolates from hospitalized patients. Pulse-field gel electrophoresis-typing yielded 63 pulsotypes, with high diversity observed in all sources (D = ≥0.901. No pulsotype was common to all the three sources. Each patient room had its own unique PFGE pattern which persisted after six months. Minimum inhibitory concentrations of Vancomycin, Gentamicin, Penicillin, Tetracycline, Nitrofurantoin, Levofloxacin, Ciprofloxacin and Fosfomycin were evaluated. Resistance to Tetracycline was most prevalent in isolates from farm animals (62% and water sources (49%. Water isolates (86% had a higher prevalence of the asa1 gene, which encodes for aggregation substance, whereas clinical (78% and farm animal isolates (87% had a higher prevalence of the esp gene, encoding a surface exposed protein. This study generates knowledge on the genetic diversity of E. faecalis with antibiotic resistance and virulence characteristics from various sources in Malaysia. Keywords: Antibiotic resistance, Enterococcus faecalis, Genetic diversity, Molecular typing, Virulence markers
Hua, Ruoyi; Xia, Yun; Wu, Wenyao; Yan, Jia; Yang, Mi
Linezolid is an oxazolidinone antibiotic commonly used to treat serious infections caused by vancomycin-resistant enterococcus. Recently, low-level linezolid resistant Enterococcus faecalis strains have emerged worldwide, but the resistant mechanisms remain undefined. Whole-transcriptome profiling was performed on an E. faecalis strain P10748 with low-level linezolid resistance in comparison with a linezolid-susceptible strain 3138 and the standard control strain ATCC29212. The functions of differentially expressed genes (DEGs) were predicted, with some DEGs potentially involved in drug resistance were validated by PCR and quantitative PCR (qPCR). RNA-Seq on three E. faecalis strains generated 1920 unigenes, with 98% of them assigned to various function groups. A total of 150 DEGs were identified in the linezolid resistant strain P10748 compared to the linezolid susceptible strains 3138 and ATCC29212. Functional analysis indicated a significant transcriptomic shift to membrane transportation and biofilm formation in strain P10748, with three significantly up-regulated DEGs predicted to be associated with drug resistance through active efflux pumps and biofilm formation. The existence of these three DEGs was further confirmed by PCR and qPCR. The significant upregulation of genes associated with efflux pumps and biofilm formation in the linezolid resistant strain suggests their roles in low-level resistance to linezolid in E. faecalis. Copyright © 2018. Published by Elsevier B.V.
Rehaiem, A; Martínez, B; Manai, M; Rodríguez, A
Characterization and purification of a bacteriocin produced by a wild Enterococcus faecium strain, isolated from a Tunisian traditional fermented milk. Enterococcus faecium MMRA was selected on the basis of its strong anti-Listeria activity. The antibacterial activity was sensitive to proteases, confirming its proteinaceous nature. It was extremely heat stable (15 min at 121 degrees C), remained active over a wide pH range (2-12), and also after treatment with lipase, amylase, organic solvents, detergents, lyophilisation and long-term storage at -20 degrees C. Production of the bacteriocin occurred throughout the logarithmic growth phase, it did not adhere to the surface of the producer cells and the mode of action was bactericidal. After partial purification of the active supernatants, a 4-kDa band with antibacterial activity was revealed by SDS-PAGE electrophoresis and bioassay. Tryptic digestion followed by MALDI-TOF mass spectrometry identified the peptide as enterocin A. The inhibitory activity of Ent. faecium MMRA, a wild strain isolated from the artisan dairy beverage 'Rayeb', is due to the synthesis of an enterocin A. Traditional fresh Tunisian fermented dairy products are generally manufactured with raw milk that can be used as a source of uncharacterized wild lactic acid bacteria strains. To our knowledge, this is the first report on the isolation of an enterocin A producing Ent. faecium from 'Rayeb'. This bacteriocin or the producing strain might have a promising potential in biopreservation to enhance the hygienic quality of this dairy product.
M. Pogány Simonová
Full Text Available Information concerning the virulence factor genes and antibiotic resistance of rabbit enterococci is limited, so in this study we tested the virulence factor genes in Enterococcus faecalis strains from rabbits. Moreover, their resistance/sensitivity to antibiotics and sensitivity to enterocins was also tested, with the aim of contributing to our enterocin spectra study and to indicate the possibility of enterocin application in prevention or contaminant elimination in rabbit husbandry. A total of 144 rabbit samples were treated using a standard microbiological method. Thirty-one pure colonies of the species Enterococcus faecalis were identified, using the MALDI-TOF identification system and confirmed using phenotyping, among which 15 strains were virulence factor gene absent. The gelE gene was the most detected (42%; however, the expression of gelatinase phenotype did not always correlate with the detection of gelE. Strains did not show ß-haemolysis and were mostly resistant to tested antibiotics, but sensitive to enterocins (Ent, mainly to Ents EK13=A (P, 2019 and Ent M. Rabbit E. faecalis strains displayed antibiotic resistant traits and the presence of expressed and silent virulence genes, but they showed high levels of sensitivity to natural antimicrobials-enterocins, which indicates the possible prevention of multidrug and virulent enterococcal contaminants by enterocins.
Chisari, Giuseppe; Chisari, Eleonora M; Borzi, Antonio M; Chisari, Clara G
Aging and oxidative stress seem to play a key role in the onset and progression of ocular surface diseases. Dry Eye Syndrome (DES) is a multifactorial disease of the tears and ocular surface in which symptoms may interfere with the ability to work and carry out daily functions. This clinical trial was a pilot study to evaluate the effects of supplementation with mixture (Saccharomyces boulardii MUCL 53837 and Enterococcus faecium LMG S-28935) on the tear film. Following the run-in period subjects were randomized in two groups: group A (n.30 subjects) and group B (n.30 subjects). Group A (control) treated only with substitute tear and group B treated with substitute tear + mixture (probiotic). The data obtained in the two study groups A and B were, respectively the following: Schirmer I: 9.2±0.2 vs. 12.8±0.4 (pSaccharomyces boulardii MUCL 53837 and Enterococcus faecium LMG S-28935) activity integration with the action of tear substitutes, along with standardization of clinical parameters of the tear film and microbiological activity in restoring of the microbiota ocular surface subject with DES. Copyright© Bentham Science Publishers; For any queries, please email at firstname.lastname@example.org.
Babar, N.; Usman, J.; Munir, T.; Gill, M. M.; Anjum, R.; Gilani, M.; Latif, M.
Objective: To determine the frequency of Vancomycin Resistant Enterococcus (VRE) in a tertiary care hospital of Rawalpindi, Pakistan. Study Design: Observational, cross-sectional study. Place and Duration of Study: Department of Microbiology, Army Medical College, Rawalpindi, from May 2011 to May 2012. Methodology: Vancomycin resistant Enterococcus isolated from the clinical specimens including blood, pus, double lumen tip, ascitic fluid, tracheal aspirate, non-directed bronchial lavage (NBL), cerebrospinal fluid (CSF), high vaginal swab (HVS) and catheter tips were cultured on blood agar and MacConkey agar, while the urine samples were grown on cystine lactose electrolyte deficient agar. Later the antimicrobial susceptibility testing of the isolates was carried out using the modified Kirby-Bauer disc diffusion method on Mueller Hinton agar. Results: A total of 190 enterococci were isolated. Of these, 22 (11.57%) were found to be resistant to vancomycin. The antimicrobial sensitivity pattern revealed maximum resistance against ampicillin (86.36%) followed by erythromycin (81.81%) and gentamicin (68.18%) while all the isolates were 100% susceptible to chloramphenicol and linezolid. Conclusion: The frequency of VRE was 11.57% with the highest susceptibility to linezolid and chloramphenicol. (author)
Iyabo C. OLADIPO
Full Text Available Cell-free extracts of six strains of Enterococcus species obtained from fermented foods were used for the green synthesis of silver nanoparticles (AgNPs, which was characterized by UV-Vis spectroscopy, Fourier-transform infrared spectroscopy (FTIR and transmission electron microscopy (TEM. The biosynthesized AgNPs were dark brown in colour having surface plasmon resonance in the range of 420-442 nm. The spherical shaped AgNPs had sizes of 4-55 nm, whose formations were facilitated by proteins as indicated by the presence of peaks 1,635-1,637 and 3,275-3,313 cm-1 in the FTIR spectra. The energy dispersive x-ray (EDX showed prominent presence of silver in the AgNPs colloidal solution, while the selected area electron diffraction was typified by the face-centred crystalline nature of silver. The particles inhibited the growth of multi-drug resistant clinical isolates of Escherichia coli, Klebsiella pneumoniae and Proteus vulgaris, and also potentiated the activities of ampicillin, ciprofloxacin and cefuroxime in the AgNPs-antibiotic synergy studies. In addition, the prospective relevance of the particles as nanopreservative in paints was demonstrated with the inhibition of growth of Staphylococcus aureus, Pseudomonas aeruginosa, Aspergillus niger and A. flavus in AgNPs-paint admixture. This report further demonstrates the green synthesis of AgNPs by strains of Enterococcus species.
Heß, Stefanie; Gallert, Claudia
Tolerance of antibiotic susceptible and antibiotic resistant Escherichia coli, Enterococcus and Staphylococcus strains from clinical and wastewater samples against ozone was tested to investigate if ozone, a strong oxidant applied for advanced wastewater treatment, will affect the release of antibiotic resistant bacteria into the aquatic environment. For this purpose, the resistance pattern against antibiotics of the mentioned isolates and their survival after exposure to 4 mg/L ozone was determined. Antibiotic resistance (AR) of the isolates was not correlating with higher tolerance against ozone. Except for ampicillin resistant E. coli strains, which showed a trend towards increased resistance, E. coli strains that were also resistant against cotrimoxazol, ciprofloxacin or a combination of the three antibiotics were similarly or less resistant against ozone than antibiotic sensitive strains. Pigment-producing Enterococcus casseliflavus and Staphylococcus aureus seemed to be more resistant against ozone than non-pigmented species of these genera. Furthermore, aggregation or biofilm formation apparently protected bacteria in subsurface layers from inactivation by ozone. The relatively large variance of tolerance against ozone may indicate that resistance to ozone inactivation most probably depends on several factors, where AR, if at all, does not play a major role.
Full Text Available Enterococcus species rarely cause bacterial meningitis without predisposing factors such as trauma, brain surgery, etc. In this study, we present a bacterial meningitis case caused by Enterococcus faecalis (E. faecalis in a 13-year-old male who was found dead at home. One hundred and forty two cm tall, 37 kg weight male had admitted to hospital two days after the beginning of complaints such as weakness, headache, swelling of left eye, nausea and vomiting. Body temperature was 37.3 oC, leucocyte count 22100/ mm3, and CRP 71 g/dl at the hospital admission. Antibiotic treatment with amoxicillin/clavulanic acid (625 mg was given to the patient but he was found dead in his house the day after. In autopsy; yellow-green purulant liquid in left frontoparietal zone, fullness of meningeal vessels and oedema was seen in brain. Isolated bacteria in cerebrospinal fluid (CSF was identificated as E. faecalis by mini API 32 Strep®. Postmortem microbiological sampling in autopsy and defining etiologic agents is important for rare meningitis cases in which antemortem identification could not be done before death.
Hllytchaikra Ferraz Fehlberg
Full Text Available The objective of this study was to standardize the high-resolution melting method for identification and discrimination of Toxoplasma gondii, Sarcocystis spp., Neospora spp., and Cryptosporidium spp. by amplification of 18S ribosomal DNA (rDNA using a single primer pair. The analyses were performed on individual reactions (containing DNA from a single species of a protozoan, on duplex reactions (containing DNA from two species of protozoa in each reaction, and on a multiplex reaction (containing DNA of four parasites in a single reaction. The proposed method allowed us to identify and discriminate the four species by analyzing the derivative, normalized, and difference melting curves, with high reproducibility among and within the experiments, as demonstrated by low coefficients of variation (less than 2.2% and 2.0%, respectively. This is the first study where this method is used for discrimination of these four species of protozoa in a single reaction.
Allegretti, L; Revolledo, L; Astolfi-Ferreira, C S; Chacón, J L; Martins, L M; Seixas, G H F; Ferreira, A J P
In Brazil, the blue-fronted Amazon parrot (Amazona aestiva) is a common pet. The faecal microbiota of these birds include a wide variety of bacterial species, the majority of which belong to the Gram-positive lactic acid bacteria (LAB) clade. The aim of this study was to investigate differences in the diversity and abundance of LAB and Bifidobacterium spp. in the cloacae between wild and captive birds and to select, identify and characterise LAB for consideration as a parrot probiotic. Cloacal swabs were collected from 26 wild and 26 captive birds. Bacterial DNA was extracted, and the 16S rRNA genes were amplified. The numbers of PCR-positive Enterococcus, Pediococcus, and Lactobacillus species isolated from wild and captive birds were significantly different (PLactobacillus, Lactococcus and Bifidobacterium. Enterococcus faecium, Pediococcus pentosaceus, Lactococcus lactis, Lactobacillus coryniformis, Lactobacillus sanfranciscensis and Bifidobacterium bifidum were the most frequently isolated species from all birds. This study increases our understanding of the faecal microbiota, and may help to improve the nutrition and habitat management of captive and wild parrots. The bacterial population identified in the faecal microbiota of clinically healthy wild and captive parrots can serve as a database to analyse variations in the gut microbiota of pathogen-infected parrots and to develop probiotics specific to these genera.
Edja Maria Melo de Brito Costa
Full Text Available INTRODUÇÃO: Muitos estudos têm sido realizados para avaliar o potencial terapêutico das plantas. OBJETIVO: Este estudo teve como objetivo avaliar a ação antimicrobiana dos extratos etanólicos da aroeira-da-praia (Schinus terebinthifolius Raddi, da aroeira-do-sertão (Astronium urundeuva [Fr. All.] Engl., da ameixa-do-mato (Ximenia americana L., da quixabeira (Syderoxylum obtusifolium [Roem et Schult.] e do hipoclorito de sódio (NaOCl a 2,5%, contra o Enterococcus faecalis (ATCC 29212. METODOLOGIA: Foi realizado teste de difusão em ágar, pelo método do poço, utilizando-se como controle positivo a clorexidina a 0,12%. Os microrganismos foram semeados em caldo BHI e incubados a 37ºC por 24 horas. Posteriormente, as suspensões microbianas foram semeadas em placas Petri, com ágar Mueller Hinton, sendo confeccionados seis poços equidistantes. As placas foram mantidas à temperatura ambiente por 2 horas, para ocorrer a pré-difusão das substâncias, e incubadas a 37ºC por 48 horas. Foram feitas as análises e medições dos halos de inibição em triplicata e os resultados foram analisados estatisticamente (teste de análise de variância [ANOVA]. RESULTADOS: A quixabeira apresentou os menores halos de inibição (teste t, p INTRODUCTION: Several studies have evaluated the therapeutic benefits of plants. OBJECTIVE: The aim of this study was to evaluate the antimicrobial activity of ethanol extracts of pepper tree (Schinus terebinthifolius Raddi, aroeira (Astronium urundeuva [Fr. All.] Engl., Olacaceae (Ximenia americana L., quixaba (Syderoxylum obtusifolium [Roem et Schult.], and sodium hypochlorite (NaOCl 2.5% against Enterococcus faecalis (ATCC 29212. METHODS: The agar diffusion test was performed and 0.12% chlorhexidine was applied as positive control. The microorganisms were allowed to grow in a brain-heart infusion broth (BHI and incubated at 37ºC for 24 hours. Subsequently, the microbial suspensions were seeded on Petri dishes
Keleher, Lauren L; Skyberg, Jerod A
Brucellosis is a globally important zoonotic infectious disease caused by gram negative bacteria of the genus Brucella. While many species of Brucella exist, Brucella melitensis, Brucella abortus, and Brucella suis are the most common pathogens of humans and livestock. The virulence of Brucella is largely influenced by its ability to evade host factors, including phagocytic killing mechanisms, which are critical for the host response to infection. The aim of this study was to characterize the bovine neutrophil response to virulent Brucella spp. Here, we found that virulent strains of smooth B. abortus, B. melitensis, B. suis, and virulent, rough, strains of Brucella canis possess similar abilities to resist killing by resting, or IFN-γ-activated, bovine neutrophils. Bovine neutrophils responded to infection with a time-dependent oxidative burst that varied little between Brucella spp. Inhibition of TAK1, or SYK kinase blunted the oxidative burst of neutrophils in response to Brucella infection. Interestingly, Brucella spp. did not induce robust death of bovine neutrophils. These results indicate that bovine neutrophils respond similarly to virulent Brucella spp. In addition, virulent Brucella spp., including naturally rough strains of B. canis, have a conserved ability to resist killing by bovine neutrophils. Copyright © 2016 Elsevier B.V. All rights reserved.
Catharina de Paula Oliveira Cavalcanti Soares
Full Text Available Objective: to determine the seroprevalence of Brucella spp in humans.Method: this is an observational study, developed with 455 individuals between 18 and 64 years old, who use the Estratégia de Saúde da Família (Brazil's family health strategy. The serum samples of volunteers underwent buffered acid antigen tests, such as screening, agar gel immunodiffusion and slow seroagglutination test in tubes and 2-Mercaptoethanol.Results: among the samples, 1.98% has responded to buffered-acid antigen, 2.85% to agar gel immunodiffusion test and 1.54% to the slow seroagglutination tests on tubes/2-Mercaptoethanol. The prevalence of Brucella spp was 4.4%, represented by the last two tests.Conclusion: the results of this research suggest that the studied population is exposed to Brucella spp infection.
H.H. Caiaffa Filho
Full Text Available Vancomycin-resistant enterococci strains (VRE is an important pathogen related with hospital infections in many countries, presenting limited or no therapeutic options for treating serious infections. VRE has presented some different genotypes been VanA and VanB considered to be the most important in hospital environments. In the present study the authors investigated the prevalence of van genes (A, B an C among clinical isolates of VRE in a five month period at a large tertiary hospital in Sao Paulo, Brazil. The results showed the presence of vanA, but not vanB or vanC in all 43 strains of E. faecalis and five E. faecium studied. The results bring an important issue, due to the possibility of resistance spread of vanA genes, to be monitored and solved by the hospital infection control team and the microbiology and molecular biology laboratories at tertiary Hospitals.
Damborg, Peter Panduro; Top, Janetta; Hendrickx, Antoni P.A.
Ampicillin resistance is a marker for hospital-associated Enterococcus faecium. Feces from 208 dogs were selectively screened for the occurrence of ampicillin-resistant E. faecium (AREF). AREF was detected in 42 (23%) of 183 dogs screened in a cross-sectional study in the United Kingdom and in 19...
E.coli and Enterococcus serve as important water quality indicator organisms. Rainfall action on manured fields and pastures releases these organisms into soil with infiltrating water. They can then be released back to runoff during subsequent rainfall or irrigation events as soil solution interacts...
Cai, J; Collins, M D
The 16S rRNA gene sequence of Melissococcus pluton, the causative agent of European foulbrood disease, was determined in order to investigate the phylogenetic relationships between this organism and other low-G + C-content gram-positive bacteria. A comparative sequence analysis revealed that M. pluton is a close phylogenetic relative of the genus Enterococcus.
Licht, Tine Rask; Hammerum, Anette Marie; Jensen, Lars Bogø
The effect of synthetic sex pheromone on pheromone-inducible conjugation between the isogenic Enterococcus faecalis strains OG1RF and OG1SS was investigated in (i) Todd-Hewitt broth medium and (ii) intestinal mucus isolated from germ-free rats. In broth, the presence of synthetic pheromone cCF10...
Scheetz, Marc H.; Knechtel, Stephanie A.; Malczynski, Michael; Postelnick, Michael J.; Qi, Chao
Clinical enterococcal resistance to linezolid is defined by the presence of the G2576T mutation. We evaluated the incidence of genetically proven linezolid resistance among vancomycin-resistant Enterococcus faecium strains and linezolid consumption for a possible association. A relationship was found (r2 = 0.73, P = 0.03) and predicts increasing resistance with current trends of linezolid use.
Bravetti, Anne-Lise; Mesnage, Stéphane; Lefort, Agnès; Chau, Françoise; Eckert, Catherine; Garry, Louis; Arthur, Michel; Fantin, Bruno
The bactericidal activity of amoxicillin was investigated against Enterococcus faecalis JH2-2 and against an isogenic mutant deficient in the production of the N-acetylglucosaminidase AtlA. Comparison of the two strains indicated that this autolysin contributes to killing by amoxicillin both in vitro and in a rabbit model of experimental endocarditis.
To the Editor:Meningitis caused by Enterococcus casseliflavus (E.casseliflavus) is extremely rare.Here we report an unusual case of meningitis caused by E.casseliflavus coexisting with refractory cerebrospinal fluid (CSF) leakage following endoscopic endonasal resection of skull base chondrosarcoma.
Sawa, Naruhiko; Wilaipun, Pongtep; Kinoshita, Seisuke; Zendo, Takeshi; Leelawatcharamas, Vichien; Nakayama, Jiro; Sonomoto, Kenji
Enterococcus faecalis NKR-4-1 isolated from pla-ra produces a novel two-peptide lantibiotic, termed enterocin W, comprising Wα and Wβ. The structure of enterocin W exhibited similarity with that of plantaricin W. The two peptides acted synergistically, and their order of binding to the cell membrane was important for their inhibitory activity.
Tedim, Ana P.; Lanza, Val F.; Manrique, Marina; Pareja, Eduardo; Ruiz-Garbajosa, Patricia; Cantón, Rafael; Baquero, Fernando; Tobes, Raquel
ABSTRACT The emergence of nosocomial infections by multidrug-resistant sequence type 117 (ST117) Enterococcus faecium has been reported in several European countries. ST117 has been detected in Spanish hospitals as one of the main causes of bloodstream infections. We analyzed genome variations of ST117 strains isolated in Madrid and describe the first ST117 closed genome sequences. PMID:28360174
Full Text Available Introduction: The present study aimed to investigate the effect of Enterococcus faecium EF55 on chickens, as well as its influence on proliferative activity of epithelial intestinal cells after infection with Salmonella enterica serovar Enteritidis phage type 4 (SE PT4. Moreover, the length and area of duodenal and jejunal villi of the birds were examined.
F.L. Paganelli; M. de Been; J.C. Braat (Johanna); T.A. Hoogenboezem (Thomas); C. Vink (Cornelis); J. Bayjanov; M.R.C. Rogers; J. Huebner; M.J.M. Bonten (Marc); R.J.L. Willems (Rob); H.L. Leavis
textabstractEnterococcus faecium is an important nosocomial pathogen causing biofilm-mediated infections. Elucidation of E. faecium biofilm pathogenesis is pivotal for the development of new strategies to treat these infections. In several bacteria, extracellular DNA (eDNA) and proteins act as
Tannock, G W
The broad-host-range plasmid pAM beta 1 (erythromycin resistance) was transferred conjugally from Streptococcus lactis to Lactobacillus reuteri, L. murinus, and L. fermentum. Transfer of pAM beta 1 between two L. reuteri strains occurred, and lactobacillus transconjugants could act as donors of pAM beta 1 in crosses with Enterococcus faecalis JH2-2.
Timmers, Gert Jan; van der Zwet, Wil C.; Simoons-Smit, Ina M.; Savelkoul, Paul H. M.; Meester, Helena H. M.; Vandenbroucke-Grauls, Christina M. J. E.; Huijgens, Peter C.
We describe an outbreak of vancomycin-resistant Enterococcus faecium (VRE) on the haematology ward of a Dutch university hospital. After the occurrence of three consecutive cases of bacteraemia with VRE, strains were genotyped and found to be identical. During the next 4 months an intensive
Hasman, Henrik; Aarestrup, Frank Møller
A significant relationship between copper resistance (tcrB), glycopeptide resistance (Tn1546), and macrolide resistance [erm(B)] in Enterococcus faecium isolated from pigs was found. The tcrB gene was located closely upstream of the Tn1546 element. However, the continued use of copper sulfate has...... not been able to maintain high levels of macrolide and glycopeptide resistance....
Madoshi, B. P.; Mtambo, M. M.A.; Muhairwa, A. P.
faecalis was the most prevalent species from all sources of isolation (43·5%), followed by Enterococcus faecium (38·4%). Isolates of E. faecium showed a higher number of phenotypic antimicrobial resistance than isolates of E. faecalis. Fifty-eight isolates, which showed resistance or intermediate...
加藤, 行男; 村上, 賢
A total of 291 fecal samples from 252 wild reptiles and 39 pet reptiles were examined for the prevalence of Salmonella spp. in Japan. Salmonella spp. were isolated from 29 (11.5%) of 252 wild reptiles and 22 (55.6%) of 39 pet reptiles. The isolates were identified into subspecies I to IV. The majority of isolates (43.6%) belonged to subspecies I and these isolates could be identified into 9 serovars. The serovars isolated were found to be S. Newport, S. Litchifield and S. Thompson which cause...
Evaluation of Enterococcus faecalis adhesion, penetration, and method to prevent the penetration of Enterococcus faecalis into root cementum: Confocal laser scanning microscope and scanning electron microscope analysis.
Halkai, Rahul S; Hegde, Mithra N; Halkai, Kiran R
To ascertain the role of Enterococcus faecalis in persistent infection and a possible method to prevent the penetration of E. faecalis into root cementum. One hundred and twenty human single-rooted extracted teeth divided into five groups. Group I (control): intact teeth, Group II: no apical treatment done, Group III divided into two subgroups. In Groups IIIa and IIIb, root apex treated with lactic acid of acidic and neutral pH, respectively. Group IV: apical root cementum exposed to lactic acid and roughened to mimic the apical resorption. Group V: apical treatment done same as Group IV and root-end filling done using mineral trioxide aggregate (MTA). Apical one-third of all samples immersed in E. faecalis broth for 8 weeks followed by bone morphogenetic protein and obturation and again immersed into broth for 8 weeks. Teeth split into two halves and observed under confocal laser scanning microscope and scanning electron microscope, organism identified by culture and polymerase chain reaction techniques. Adhesion and penetration was observed in Group IIIa and Group IV. Only adhesion in Group II and IIIB and no adhesion and penetration in Group I and V. Adhesion and penetration of E. faecalis into root cementum providing a long-term nidus for subsequent infection are the possible reason for persistent infection and root-end filling with MTA prevents the adhesion and penetration.
Donna M. Ferguson
Full Text Available Most Enterococcus faecalis and E. faecium are harmless to humans; however, strains harboring virulence genes, including esp, gelE, cylA, asa1, and hyl, have been associated with human infections. E. faecalis and E. faecium are present in beach waters worldwide, yet little is known about their virulence potential. Here, multiplex PCR was used to compare the distribution of virulence genes among E. faecalis and E. faecium isolated from beaches in Southern California and Puerto Rico to isolates from potential sources including humans, animals, birds, and plants. All five virulence genes were found in E. faecalis and E. faecium from beach water, mostly among E. faecalis. gelE was the most common among isolates from all source types. There was a lower incidence of asa1, esp, cylA, and hyl genes among isolates from beach water, sewage, septage, urban runoff, sea wrack, and eelgrass as compared to human isolates, indicating that virulent strains of E. faecalis and E. faecium may not be widely disseminated at beaches. A higher frequency of asa1 and esp among E. faecalis from dogs and of asa1 among birds (mostly seagull suggests that further studies on the distribution and virulence potential of strains carrying these genes may be warranted.
Full Text Available Objective: To do surveillance for vancomycin and inducible clindamycin resistance of Enterococcus faecalis (E. faecalis, a Gram-positive bacterium in a teaching hospital. Methods: E. faecalis strains isolated from clinical samples were screened for vancomycin and inducible clindamycin resistance, i.e., D-test positivity, using vancomycin screen agar and blood agar plates, respectively. For the D-test screening, erythromycin resistant (Er-r and clindamycin sensitive (Cd-s strain were used. Results: Of 265 isolated E. faecalis strains, 159 (60% were vancomycin resistant Enterococcus (VRE and 106 were vancomycin sensitive Enterococcus (VSE. Of 265 strains, 42 were constitutively resistant to clindamycin and erythromycin and of 148 Er-r and Cd-s strains, 87 (32.83% had D-test positivity, while the rest 61 strains were D-test negatives. D-test results examined with 6 hospital factors as bivalents, only 2 factors, the VSE/VRE and the presence/absence of prior antibiotic use > 90 days bivalent were statistically significant. A VRE strain with D-test positivity would be picked up 0.570 2 times more frequently than a strain with VSE and D-test positivity. Also, patients with prior antibiotic use > 90 days had 3.737 5 times more chance of picking up D-test positive strains than patients without any prior antibiotic use. Resistance pattern of E. faecalis strains to individual 14 antibiotics were recorded; the maximum values of resistance were against ampicillin 10 μg/disc and linezolid 30 μg/disc. Student’s t-test for hospital acquired and community acquired data revealed that drug resistant strains were equally prevalent in both sources. Conclusions: Prevalence of 60% VRE in both hospital and adjoining community creates consternation. In total 87 (32.83% strains had D-test positivity; patients who had used antibiotics within the last 90 days have got an ample chance of picking of D-test positive E. faecalis. D-test protocol should be followed with
... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3630 Serratia spp... antigens and antisera used in serological tests to identify Serratia spp. from cultured isolates. The...
... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3660 Shigella spp...), used in serological tests to identify Shigella spp. from cultured isolates. The identification aids in...
... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3035 Arizona spp... antisera and antigens used to identify Arizona spp. in cultured isolates derived from clinical specimens...
Gisbert Domenech, Maria Carmen; Picó Sirvent, María Belén N:2949; Nuez Viñals, Fernando
Gisbert Domenech, MC.; Picó Sirvent, MBN.; Nuez Viñals, F. (2011). Regeneration in selected Cucurbita spp. germplasm. Report- Cucurbit Genetics Cooperative. 33-34:53-54. http://hdl.handle.net/10251/62926 Senia 53 54 33-34
Full Text Available Malaria remains a public health problem in West Nusa Tenggara Province. Central Lombok District is one of the areas with high case of malaria. Annual Malaria Incidence (AMI was increased from 5.9 ‰ in 2006, 6.7‰ up to 8.1‰ in 2008. The objective of the study is to describe the ecological condition of Anopheles spp. through observation, measurement of environmental variables, larvae and adult collection. This research was an observational research with cross-sectional study. The population of this study is all mosquitos and breeding habitats of Anopheles spp. that exist in the research location. Ecological observations carried out on anopheles breeding habitats including acidity, salinity, shaded places and aquatic biota. Air temperature and humidity measured at the adult mosquitoes trapping sites. The result showed that pH values of water is around 9.00, salinity in the breeding habitats around 14 ppm, and water biota (i.e. moss, grass, shrimps, fishes, tadpoles and crabs surrounded by bushes with larvae density 0,1-28,8 each dipping. The air measurement at the time was between 23°-27° Celsius and 65%-84% humidity. This research concludes that ecology and environmental conditions were supporting the development of larvae and adult mosquito of Anopheles spp.Keywords: ecology, Anopheles spp., Central Lombok
Hewlett, T. E.; Cox, R.; Dickson, D. W.; Dunn, R. A.
Two years of data collected from the Florida Nematode Assay Laboratory of the Florida Cooperative Extension Service and 4 years of data from the Florida Department of Agriculture and Consumer Services, Division of Plant Industry, were compiled to find out the distribution of Pasteuria spp. on nematodes in Florida soils. Information recorded came from 335 samples and included nematode genera with Pasteuria endospores attached, host plants associated with the samples, and the origins of the samples. Pasteuria spp. were detected on 14 different plant-parasitic nematode genera in 41 Florida counties and associated with over 39 different plant species and in seven fallow fields. Pasteuria-infected nematodes were associated with a wide range of plant hosts, although frequency of associations with these hosts reflected the sample bias of the laboratories involved. Meloidogyne and Hoplolaimus spp. were the two nematode genera most frequently associated with Pasteuria. Pasteuria spp. were observed attached to members of these two genera in 176 and 59 soil samples, respectively. PMID:19279936
Hochel, I.; Růžičková, H.; Krásný, Lukáš; Demnerová, K.
Roč. 112, č. 6 (2012), s. 1257-1265 ISSN 1364-5072 Grant - others:GA ČR(CZ) GAP503/10/0664 Institutional research plan: CEZ:AV0Z50200510 Keywords : biotype * antibiotic resistance * Cronobacter spp Subject RIV: CE - Biochemistry Impact factor: 2.196, year: 2012
Durum wheat is the major cereal crop cultivated in Tunisia; covering over 40% of the cereal growing areas. Durum wheat production remains below expectation due to its low productivity that is attributed to the chronically abiotic and biotic stresses. Fusarium head blight (FHB) caused by Fusarium spp. has become an ...
Ordeix, Laura; Galeotti, Franca; Scarampella, Fabia; Dedola, Carla; Bardagí, Mar; Romano, Erica; Fondati, Alessandra
A series of 18 allergic cats with multifocal Malassezia spp. overgrowth is reported: atopic dermatitis was diagnosed in 16, an adverse food reaction in another and one was euthanized 2 months after diagnosis of Malassezia overgrowth. All the cats were otherwise healthy and those tested (16 out of 18) for feline leukaemia or feline immunodeficiency virus infections were all negative. At dermatological examination, multifocal alopecia, erythema, crusting and greasy adherent brownish scales were variably distributed on all cats. Cytological examination revealed Malassezia spp. overgrowth with/without bacterial infection in facial skin (n = 11), ventral neck (n = 6), abdomen (n = 6), ear canal (n = 4), chin (n = 2), ear pinnae (n = 2), interdigital (n = 1) and claw folds skin (n = 1). Moreover, in two cats Malassezia pachydermatis was isolated in fungal cultures from lesional skin. Azoles therapy alone was prescribed in seven, azoles and antibacterial therapy in eight and azoles with both antibacterial and anti-inflammatory therapy in three of the cats. After 3-4 weeks of treatment, substantial reduction of pruritus and skin lesions was observed in all 11 cats treated with a combined therapy and in five of seven treated solely with azoles. Malassezia spp. overgrowth may represent a secondary cutaneous problem in allergic cats particularly in those presented for dermatological examination displaying greasy adherent brownish scales. The favourable response to treatment with antifungal treatments alone suggests that, as in dogs, Malassezia spp. may be partly responsible for both pruritus and cutaneous lesions in allergic cats.
... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Echinococcus spp. serological reagents. 866.3200... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3200 Echinococcus spp. serological reagents. (a) Identification. Echinococcus spp. serological reagents are devices that...
Verdejo Lucas, Soledad; Español Pons, Montserrat; Ornat Longarón, Cèsar; Sorribas Royo, Francisco Javier
The occurrence of Pasteuria spp. In Spanish oils is reported. A total of 160 soil samples were collected from vegetable crops, kiwi and citrus orchards, and deciduous fruit trees. Bacteria were found associated with six nematode genera but they were only observed within females of Meloidogyne spp., second-stage juveniles and males of Tylenchulus semipenetrans, and juveniles of Pratylenchus spp.
... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Pseudomonas spp. serological reagents. 866.3415... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3415 Pseudomonas spp. serological reagents. (a) Identification. Pseudomonas spp. serological reagents are devices that...
... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Leptospira spp. serological reagents. 866.3350... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3350 Leptospira spp. serological reagents. (a) Identification. Leptospira spp. serological reagents are devices that...
... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3085 Brucella spp... from clinical specimens or to identify antibodies to Brucella spp. in serum. Additionally, some of... to identify Brucella spp. directly from clinical specimens or cultured isolates derived from clinical...
M’ikanatha, Nkuchia M.; Dettinger, Lisa A.; Perry, Amanda; Rogers, Paul; Reynolds, Stanley M.
In 2010, we surveyed 176 clinical laboratories in Pennsylvania regarding stool specimen testing practices for enteropathogens, including Campylobacter spp. Most (96.3%) routinely test for Campylobacter spp. In 17 (15.7%), a stool antigen test is the sole method for diagnosis. We recommend that laboratory practice guidelines for Campylobacter spp. testing be developed. PMID:22377086
The spread of introduced saltcedar (Tamarix spp.) throughout many riparian systems across the western United States motivated the introduction of biological control agents that are specific to saltcedar, saltcedar leaf beetles (Diorhabda carinulata, D. elongata; Chrysomelidae). I monitored small mam...
Our objective was to determine the suppressive abilities of bacterial metabolites derived from Photorhabdus and Xenorhabdus spp. on Glomerella cingulata, Phomopsis sp., Phytophthora cactorum, and Fusicladosporium effusum, which are fungal or oomycete pathogens of pecan, and Monilinia fructicola, a f...
Jensen, Lars Bogø; Hammerum, Anette Marie; Bager, Flemming
The genetic background for streptogramin resistance was examined in Enterococcus faecium isolated from pigs (n = 55) and broilers (n = 207) in 1997 in Denmark. Fifty-one percent and 67%, respectively, of the isolates were resistant to streptogramins. Among streptogramin-resistant E. faecium (SREF......), the genetic background for streptogramin A resistance could be determined in 96% of the isolates from broilers, compared with 14% among SREF from pigs. For broiler isolates 89% of SREF contained the vat(E) gene and 10% the vat(D) gene. Three of these isolates contained both resistance genes. Among SREF from...... pigs two isolates contained the vat(E) gene and two others the vat(D) gene. The genetic background for streptogramin B was most often identified as the erm(B) gene encoding macrolide, lincosamide, and streptogramin B (MLSB) resistance. Among SREF, 84% and 86% of isolates from broilers and pigs...
Aarestrup, Frank Møller; McNicholas, P. M.
Six high-level evernimicin-resistant Enterococcus faecium isolates were identified among 304 avilarnycin-resistant E. faecium isolates from animals and 404 stool samples from humans with diarrhea. All four animal isolates, and one of the human isolates, were able to transfer resistance...... to a susceptible E. faecium strain. The resulting transconjugants all tested positive for the presence of emtA, a gene encoding a methyltransferase previously linked with high-level evernimicin resistance. The four transconjugants derived from animal isolates all carried the same plasmid, while a differently sized...... plasmid was found in the isolate from humans. This study demonstrated a low incidence of high-level evernimicin resistance mediated by the emtA gene in different E. faecium isolates of animal and human origin....
Shankar, Nathan; Baghdayan, Arto S.; Willems, Rob
Enterococcus faecalis isolates of porcine origin were screened for the presence of a previously identified pathogenicity island (PAI). By using the esp gene as a genetic marker for the presence of this PAI, 9 esp-positive and 10 esp-negative isolates of porcine origin were investigated by use...... of a designed oligonucleotide array. The results indicated the clustering of esp-positive strains by multilocus sequence typing (MLST), but surprisingly, all strains investigated contained parts of the PAL None of the strains of animal origin investigated belonged to previously identified MLST complex 2, where...... most isolates from patients cluster. Five of the nine esp-positive E. faecalis isolates of animal origin belonged to the same PAI complex as human isolate MMH594 but differed in their sequence types, which strongly indicates the horizontal transfer of the PAI between enterococci of porcine and human...
Meining, Winfried; Scheuring, Johannes; Fischer, Markus; Weinkauf, Sevil
SecA ATPase from E. faecalis has been cloned, overexpressed, purified and crystallized. Crystals belong to space group C2 and diffract to 2.4 Å resolution. The gene coding for SecA from Enterococcus faecalis was cloned and overexpressed in Escherichia coli. In this protein, the lysine at position 6 was replaced by an asparagine in order to reduce sensitivity towards proteases. The modified protein was purified and crystallized. Crystals diffracting to 2.4 Å resolution were obtained using the vapour-diffusion technique. The crystals belong to the monoclinic space group C2, with unit-cell parameters a = 203.4, b = 49.8, c = 100.8 Å, α = γ = 90.0, β = 119.1°. A selenomethionine derivative was prepared and is currently being tested in crystallization trials
Fabrício Luiz Tulini
Full Text Available Lactic acid bacteria are important in foods as potential probiotics and also due to the ability to produce antimicrobial compounds that can contribute for biopreservation. In this work, the bacteriocin produced by the food isolate Enterococcus faecium 130 was partially purified and characterized. The compound was active against Gram-positive bacteria, including Listeria monocytogenes. It was produced after 4 days of storage at a broad temperature range (4 to 37 °C; it was stable at pH ranging from 2 to 10 with no loss of activity after heating at 100 °C for 15 minutes. Bacteriocin was partially purified by the adsorption-desorption technique, and the analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE showed a molecular mass of 3.5 to 6.5 kDa. These data encourage studies on application of this bacteriocin in food systems as an additional hurdle to microbial growth.
Emilly Caroline de Freitas Silva
Full Text Available Introduction. Enterococcus faecalis (E. faecalis, a constituent of the gut microbiota, can be associated with both colonic lesions and endocarditis. Since this microorganism is one of the endocarditis etiological agents, there is a need for greater study in regard to the association with endocarditis and colonic lesions. Case Presentation. This is the case description of a 53-year-old man with history of prolapse of the anterior mitral valve leaflet who was diagnosed with endocarditis by E. faecalis and treated with ampicillin and gentamicin. Upon investigation by colonoscopy, he was found to have a tubular adenoma with low grade dysplasia. Conclusion. There are a few descriptions in scientific literature of an association between endocarditis by E. faecalis and colonic lesions. However, further studies with significant correlation between the two pathologies are required, so that proper measures can be implemented in clinical practice.
Green, M; Barbadora, K; Wadowsky, R M
The combination of ampicillin and vancomycin kills some but not all strains of ampicillin- and vancomycin-resistant Enterococcus faecium. We compared a simple test for synergy utilizing a commercially available microdilution susceptibility system with time-kill studies and determined acceptable breakpoints for this test for 20 strains of ampicillin- and vancomycin-resistant E. faecium. The combination of ampicillin and vancomycin was tested for synergy by time-kill, broth macrodilution, and broth microdilution procedures. Repeat testing of isolates by macro- and microdilution synergy methods yielded MICs that were within one twofold dilution of each other for both intra- and intertest comparisons. Synergy was always detected by time-kill studies when the MIC of ampicillin in the combination synergy screen was 16 micrograms/ml in the combination microdilution synergy screen. The determination of the synergy by the broth microdilution procedure appears to be simple, convenient, and accurate.
Full Text Available CRISPR-Cas systems, which obstruct both viral infection and incorporation of mobile genetic elements by horizontal transfer, are a specific immune response common to prokaryotes. Antiviral protection by CRISPR-Cas comes at a cost, as horizontally-acquired genes may increase fitness and provide rapid adaptation to habitat change. To date, investigations into the prevalence of CRISPR have primarily focused on pathogenic and clinical bacteria, while less is known about CRISPR dynamics in commensal and environmental species. We designed PCR primers and coupled these with DNA sequencing of products to detect and characterize the presence of cas1, a universal CRISPR-associated gene and proxy for the Type II CRISPR1-Cas system, in environmental and non-clinical Enterococcus isolates. CRISPR1-cas1 was detected in approximately 33% of the 275 strains examined, and differences in CRISPR1 carriage between species was significant. Incidence of cas1 in E. hirae was 73%, nearly three times that of E. faecalis (23.6% and 10 times more frequent than in E. durans (7.1%. Also, this is the first report of CRISPR1 presence in E. durans, as well as in the plant-associated species E. casseliflavus and E. sulfureus. Significant differences in CRISPR1-cas1 incidence among Enterococcus species support the hypothesis that there is a tradeoff between protection and adaptability. The differences in the habitats of enterococcal species may exert varying selective pressure that results in a species-dependent distribution of CRISPR-Cas systems.
Nami, Y; Abdullah, N; Haghshenas, B; Radiah, D; Rosli, R; Yari Khosroushahi, A
This study aimed to describe probiotic properties and bio-therapeutic effects of newly isolated Enterococcus faecalis from the human vaginal tract. The Enterococcus faecalis strain was originally isolated from the vaginal microbiota of Iranian women and was molecularly identified using 16SrDNA gene sequencing. Some biochemical methodologies were preliminarily used to characterize the probiotic potential of Ent. faecalis, including antibiotic susceptibility, antimicrobial activity, as well as acid and bile resistance. The bio-therapeutic effects of this strain's secreted metabolites on four human cancer cell lines (AGS, HeLa, MCF-7 and HT-29) and one normal cell line (HUVEC) were evaluated by cytotoxicity assay and apoptosis scrutiny. The characterization results demonstrated into the isolated bacteria strain revealed probiotic properties, such as antibiotic susceptibility, antimicrobial activity and resistance under conditions similar to those in the gastrointestinal tract. Results of bio-therapeutic efficacy assessments illustrated acceptable apoptotic effects on four human cancer cell lines and negligible side effects on assayed normal cell line. Our findings revealed that the apoptotic effect of secreted metabolites mainly depended on proteins secreted by Ent. faecalis on different cancer cells. These proteins can induce the apoptosis of cancer cells. The metabolites produced by this vaginal Ent. faecalis strain can be used as alternative pharmaceutical compounds with promising therapeutic indices because they are not cytotoxic to normal mammalian cells. Accordingly, the physicochemical, structural and functional properties of the secreted anticancer substances should be further investigated before using them as anticancer therapeutics. This study aim to screen total bacterial secreted metabolites as a wealthy source to find the new active compounds to introduce as anticancer therapeutics in the future. © 2014 The Society for Applied Microbiology.
Franz, Charles M. A. P.; Worobo, Randy W.; Quadri, Luis E. N.; Schillinger, Ulrich; Holzapfel, Wilhelm H.; Vederas, John C.; Stiles, Michael E.
A purified bacteriocin produced by Enterococcus faecium BFE 900 isolated from black olives was shown by Edman degradation and mass spectrometric analyses to be identical to enterocin B produced by E. faecium T136 from meat (P. Casaus, T. Nilsen, L. M. Cintas, I. F. Nes, P. E. Hernández, and H. Holo, Microbiology 143:2287–2294, 1997). The structural gene was located on a 2.2-kb HindIII fragment and a 12.0-kb EcoRI chromosomal fragment. The genetic characteristics and production of EntB by E. faecium BFE 900 differed from that described so far by the presence of a conserved sequence like a regulatory box upstream of the EntB gene, and its production was constitutive and not regulated. The 2.2-kb chromosomal fragment contained the hitherto undetected immunity gene for EntB in an atypical orientation that is the reverse of that of the structural gene. Typical transport and other genes associated with bacteriocin production were not detected on the 12.0-kb chromosomal fragment containing the EntB structural gene. This makes the EntB genetic system different from most other bacteriocin systems, where transport and possible regulatory genes are clustered. EntB was subcloned and expressed by the dedicated secretion machinery of Carnobacterium piscicola LV17A. The structural gene was amplified by PCR, fused to the divergicin A signal peptide, and expressed by the general secretory pathway in Enterococcus faecalis ATCC 19433. PMID:10224016
Martins Clélia Aparecida de Paiva
Full Text Available The presence of yeasts and staphylococci in the oral cavity is important because they can act as supplementary microbiota and in certain situations can cause oral or systemic diseases. The aim of this work was to study the prevalence of Candida spp. and Staphylococcus spp. in the human oral cavity. Oral rinses were collected from sixty-eight individuals according to the technique described by Samaranayake and MacFarlane and then cultured on Sabouraud medium supplemented with chloramphenicol and Baird-Parker agar. After the incubation period, the microorganisms were isolated and identified through biochemical tests. The data obtained were statistically analysed by ANOVA. Candida spp. were isolated from 61.76% of the examined individuals and C. albicans was the more frequently isolated specie. Staphylococcus spp. were isolated from 95.60% of the individuals and 41 strains were coagulase negative (63%. Among the coagulase positive strains, nine were S. aureus, 11 S. hyicus and 4 S. schleiferi subspecie coagulans. No correlation was observed between the counts (cfu of the isolated Candida spp. and Staphylococcus spp.
Guerra, Diogo Ribeiro Almeida
Dissertação de Mestrado Integrado em Medicina Veterinária Echinococcus spp., Taenia spp. and Toxocara spp. are important parasites of domestic and wild canids and neglected zoonotic helminths. Despite their relevance in Public Health, little is known about their prevalence in Portugal. An epidemiological study was conducted to clarify the role of canids in the sylvatic and synanthropic cycles of these pathogens in our country. Fecal samples from dog (n = 51), red fox (n = 62) and Iberia...
Bialvaei, Abed Zahedi; Kafil, Hossein Samadi; Asgharzadeh, Mohammad; Aghazadeh, Mohammad; Yousefi, Mehdi
This study was conducted in Iran in order to assess the distribution of CTX-M type ESBLs producing Enterobacteriaceae. From January 2012 to December 2013, totally 198 E. coli, 139 Klebsiella spp, 54 Salmonella spp and 52 Shigella spp from seven hospitals of six provinces in Iran were screened for resistance to extended-spectrum cephalosporins. After identification and susceptibility testing, isolates presenting multiple-drug resistance (MDR) were evaluated for ESBL production by the disk combination method and by Etest using (cefotaxime and cefotaxime plus clavulanic acid). All isolates were also screened for blaCTX-M using conventional PCR. A total of 42.92%, 33.81%, 14.81% and 7.69% of the E. coli, Klebsiella spp, Salmonella spp and Shigella spp isolates were MDR, respectively. The presence of CTX-M enzyme among ESBL-producing isolates was 85.18%, 77.7%, 50%, and 66.7%, in E. coli, Klebsiella spp, Salmonella spp and Shigella spp respectively. The overall presence of CTX-M genes in Enterobacteriaceae was 15.4% and among the resistant isolates was 47.6%. This study indicated that resistance to β-lactams mediated by CTX-M enzymes in Iran had similar pattern as in other parts of the world. In order to control the spread of resistance, comprehensive studies and programs are needed. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.
Paola Mendes Milanesi; Elena Blume; Marlove Fátima Brião Muniz; Lia Rejane Silveira Reiniger; Zaida Inês Antoniolli; Emanuele Junges; Manoeli Lupatini
This study aimed i) to quantify the occurrence of Fusarium spp. and Trichoderma spp. in rhizospheric soil, with and without symptoms of Sudden Death Syndrome (SDS) in eight soybean genotypes; ii) morphologically identify isolates of Fusarium spp. from roots with SDS; iii) evaluate the antagonism between Trichoderma spp. and Fusarium spp. isolates from rhizospheric soil and roots from with and without SDS, respectively; and iv) characterize through the ITS1-5.8S-ITS2 region of rDNA the isolate...
Skinner, Delaina; Mitcham, Jessica R; Starkey, Lindsay A; Noden, Bruce H; Fairbanks, W Sue; Little, Susan E
American black bears (Ursus americanus) are commonly infested with ticks throughout their range, but there are few surveys for tick-borne disease agents in bears. To characterize tick infestations and determine the prevalence of current infection with Babesia spp. and past or current infection with Ehrlichia spp. in newly re-established populations of black bears in east central and southeastern Oklahoma, US, we identified adult (n=1,048) and immature (n=107) ticks recovered from bears (n=62). We evaluated serum and whole blood samples from a subset (n=49) for antibodies reactive to, and characteristic DNA fragments of, Ehrlichia spp., as well as characteristic DNA fragments of Babesia spp. Amblyomma americanum, the most common tick identified, was found on a majority (56/62; 90%) of bears and accounted for 697/1,048 (66.5%) of all ticks recovered. Other ticks included Dermacentor variabilis (338/1,048; 32.3%) from 36 bears, Amblyomma maculatum (9/1,048; 0.9%) from three bears, and Ixodes scapularis (4/1,048; 0.4%) from three bears. Antibodies reactive to Ehrlichia spp. were detected in every bear tested (49/49; 100%); maximum inverse titers to Ehrlichia chaffeensis ranged from 64-4,096 (geometric mean titer 1,525). However, PCR failed to identify active infection with E. chaffeensis, Ehrlichia ewingii, or an Ehrlichia ruminantium-like agent. Infection with Babesia spp. was detected by PCR in 3/49 (6%) bears. Together these data confirm that tick infestations and infection with tick-borne disease agents are common in bears in the southern US. The significance of these infestations and infections to the health of bears, if any, and the identity of the Ehrlichia spp. responsible for the antibody reactivity seen, warrant further evaluation.
Landmeyer, James E.; Garigen, Thomas J.
Bacteria related to the intestinal tract of humans and other warm-blooded animals have been detected in fresh and saline surface waters used for recreational purposes in coastal areas of Horry County, South Carolina, since the early 2000s. Specifically, concentrations of the facultative anaerobic organism, Enterococcus, have been observed to exceed the single-sample regulatory limit of 104 colony forming units per 100 milliliters of water. Water bodies characterized by these concentrations are identified on the 303(d) list for impaired water in South Carolina; moreover, because current analytical methods used to monitor Enterococcus concentrations take up to 1 day for results to become available, water-quality advisories are not reflective of the actual health risk.
B. Riet-Correa; M.B. Castro; R.A. Lemos; G. Riet-Correa; V. Mustafa; F. Riet-Correa
Brachiaria species are the most important grasses for cattle production in Brazil. However, a limiting factor for the use of Brachiaria spp. is their toxicity. Most outbreaks of hepatogenous photosensitization are caused by B. decumbens; however B. brizantha, B. humidicola and B. ruziziensis can also cause poisoning. The poisoning affects cattle, sheep, goats and buffalo. Sheep are more susceptible than other animal species and the young are more susceptible than adults. There are differences...
do Amaral, Renan Bressianini; Lourenço, Elizabete Captivo; Famadas, Kátia Maria; Garcia, Amanda Barbosa; Machado, Rosangela Zacarias; André, Marcos Rogério
The family Streblidae comprises a monophyletic group of Hippoboscoidea, hematophagous dipterans that parasitize bats. Bartonella spp. and Rickettsia spp. have been reported in bats sampled in Europe, Africa, Asia, North, Central and South America. However, there are few reports on the Bartonella and Rickettsia bacteria infecting Hippoboscoidea flies and mites. While Spinturnicidae mites are ectoparasites found only in bats, those belonging to the family Macronyssidae comprise mites that also parasitize other mammal species. This study investigates the occurrence and assesses the phylogenetic positioning of Bartonella spp. and Rickettsia spp. found in Streblidae flies and Spinturnicidae and Macronyssidae mites collected from bats captured in Brazil. From May 2011 to April 2012 and September 2013 to December 2014, 400 Streblidae flies, 100 Macronyssidaes, and 100 Spinturnicidae mites were collected from bats captured in two sites in northeastern Nova Iguaçu, Rio de Janeiro, southeastern Brazil. Forty (19.8%) out of 202 Streblidae flies were positive for Bartonella spp. in qPCR assays based on the nuoG gene. Among the flies positive for the bacterium, six (18%) were Paratrichobius longicrus, seven (29%) Strebla guajiro, two (40%) Aspidoptera phyllostomatis, five (11%) Aspidoptera falcata, one (10%) Trichobius anducei, one (25%) Megistopoda aranea, and 18 (32%) Trichobius joblingi, and collected from bats of the following species: Artibeus lituratus, Carollia perspicillata, Artibeus planirostris, Sturnira lilium, and Artibeus obscurus. Six sequences were obtained for Bartonella (nuoG [n = 2], gltA [n = 2], rpoB [n = 1], ribC = 1]). The phylogenetic analysis based on gltA (750pb) gene showed that the Bartonella sequences clustered with Bartonella genotypes detected in bats and ectoparasites previously sampled in Latin America, including Brazil. Only one sample (0.49%) of the species Trichobius joblingi collected from a specimen of Carollia perspicillata was positive
Dziedzic, Ewa; Jagła, Joanna
Micropropagation is the most appropriate method for large-scale production of Rubus and Ribes spp. The proliferation rate of Rubus spp. differs in shoot tips and nodal segments. The culture media used for raspberry and blackberry propagation are MS-based supplemented with different combination and ratio of plant growth regulators, depending on the stage of culture. The initiation medium containing 0.4 mg L(-1) BA and 0.1 mg L(-1) IBA is used to stabilize shoot cultures. In multiplication media, concentration of cytokinin is doubled. In vitro rooting of shoots is achieved on media supplemented with 1.0 mg L(-1) IBA. Ribes spp. cultures are initiated from shoot tips, meristem, or dormant buds on MS medium supplemented with 2.0 mg L(-1) BA, 0.5 mg L(-1) IBA, and 0.1 mg L(-1) GA(3.) After stabilization of shoot cultures in 3-4-week time, shoot multiplication is carried out on MS medium containing 1.0 mg L(-1) BA and 0.1 mg L(-1) IBA. Shoots 2 cm long are cultured to rooting on a medium amended with 2.0 mg L(-1) IBA and 5.0 mg L(-1) IAA. Rooted plantlets are transferred to universal peat substrate and acclimatized in the greenhouse.
Sedlata Juraskova, E; Sedlackova, H; Janska, J; Holy, O; Lalova, I; Matouskova, I
To determine the current presence of Legionella spp. in the output water of dental unit waterlines (DUWLs) and examine its mitigation by disinfection at the Institute of Dentistry and Oral Sciences, Faculty of Medicine and Dentistry, Palacky University Olomouc and University Hospital Olomouc. The first stage of our survey involved collecting samples of DUWL output water from 50 dental chair units (DCUs), and 2 samples of the incoming potable water. In October 2015, a one-time disinfection (1 % Stabimed) of DUWLs was conducted. This was followed by collecting 10 control samples (survey stage 2). From the total of 50 samples (survey stage 1), 18 samples (36.0 %) tested positive for Legionella spp. Following the disinfection, nine of the ten samples no longer showed any presence of Legionella. Based on culture results, the one-time disinfection (1 % Stabimed) was effective. We are unable to comment on the duration of positive effect of disinfection on the occurrence of Legionella spp. in the outlet water. It was a one-time survey (Tab. 2, Ref. 32).
Carvalheira, Ana; Ferreira, Vânia; Silva, Joana; Teixeira, Paula
Relatively little is known about the role of foods in the chain of transmission of acinetobacters and the occurrence of different Acinetobacter spp. in foods. Currently, there is no standard procedure to recover acinetobacters from food in order to gain insight into the food-related ecology and epidemiology of acinetobacters. This study aimed to assess whether enrichment in Dijkshoorn enrichment medium followed by plating in CHROMagar™ Acinetobacter medium is a useful method for the isolation of Acinetobacter spp. from foods. Recovery of six Acinetobacter species from food spiked with these organisms was compared for two selective enrichment media (Baumann's enrichment and Dijkshoorn's enrichment). Significantly (p enrichment. Next, the Dijkshoorn's enrichment followed by direct plating on CHROMagar™ Acinetobacter was applied to detect Acinetobacter spp. in different foods. Fourteen different presumptive acinetobacters were recovered and assumed to represent nine different strains on the basis of REP-PCR typing. Eight of these strains were identified by rpoB gene analysis as belonging to the species Acinetobacter johnsonii, Acinetobacter calcoaceticus, Acinetobacter guillouiae and Acinetobacter gandensis. It was not possible to identify the species level of one strain which may suggests that it represents a distinct species. Copyright © 2015 Elsevier Ltd. All rights reserved.
Fang, Chong; Stiegeler, Emanuel; Cook, Gregory M.; Mascher, Thorsten; Gebhard, Susanne
To combat antibiotic resistance of Enterococcus faecalis, a better understanding of the molecular mechanisms, particularly of antibiotic detection, signal transduction and gene regulation is needed. Because molecular studies in this bacterium can be challenging, we aimed at exploiting the genetically highly tractable Gram-positive model organism Bacillus subtilis as a heterologous host. Two fundamentally different regulators of E. faecalis resistance against cell wall antibiotics, the bacitra...
Price, Valerie J.; Huo, Wenwen; Sharifi, Ardalan; Palmer, Kelli L.
ABSTRACT Enterococcus faecalis is an opportunistic pathogen and a leading cause of nosocomial infections. Conjugative pheromone-responsive plasmids are narrow-host-range mobile genetic elements (MGEs) that are rapid disseminators of antibiotic resistance in the faecalis species. Clustered regularly interspaced short palindromic repeat (CRISPR)-Cas and restriction-modification confer acquired and innate immunity, respectively, against MGE acquisition in bacteria. Most multidrug-resistant E.?fa...
Li, Bin; Ma, Chuan-Ling; Yu, Xiao; Sun, Yao; Li, Mei-Mei; Ye, Jian-Zhong; Zhang, Ya-Pei; Wu, Qing; Zhou, Tie-Li
The epidemiological and molecular characteristics of eight linezolid nonsusceptible Enterococcus faecalis isolated from a teaching hospital in China (January to July 2014) were investigated. The target site modifications and cfr gene associated with linezolid resistance were not found. Results of the epidemiological investigation indicated that linezolid resistance possibly occurred on several independent occasions and was often not related to linezolid administration. Copyright © 2015. Published by Elsevier B.V.
Liu, Wenli; Zhang, Lanwei; Yi, Huaxi
The macro- and micronutrients in traditional medium, such as MRS, used for cultivating lactic acid bacteria, especially for bacteriocin production, have not been defined, preventing the quantitative monitoring of metabolic flux during bacteriocin biosynthesis. To enhance Enterocin Y31 production and simplify steps of separation and purification, we developed a simplified chemically defined medium (SDM) for the growth of Enterococcus faecium Y31 and production of its bacteriocin, Enterocin Y31...
Full Text Available El diacetilo es un compuesto aromático esencial en productos lácteos fermentados como el queso. En este trabajo se estudiaron características cinéticas y bioquímicas de la α-acetolactato sintetasa (α-ALS y su influencia en la producción de diacetilo en Enterococcus faecium ETw7 y Enteroccoccus faecalis ETw23. En ambos casos, los parámetros cinéticos revelaron una baja afinidad por el piruvato, como ha sido descrito en otras bacterias ácido lácticas. E. faecium ETw7 desarrolló la máxima actividad enzimática a pH 5,8—6,2 y 40 ºC, sin embargo bajo las condiciones de maduración de quesos (pH 5,0 y 15 oC la actividad remanente fue baja. La α-ALS de E. faecalis ETw23 mostró la máxima actividad al pH de maduración, la temperatura óptima fue determinada a 40 ºC y la actividad remanente a 15 ºC fue aproximadamente el 30% de la máxima. El crecimiento y la producción de diacetilo fue estudiada en el medio De Man-Rogosa-Sharpe (MRS y MRS suplementado con citrato (MRScit. La tasa de crecimiento de E. faecium ETw7 fue comparable en ambos medios, pero se observó un aumento de la biomasa en MRScit. En el caso de E. faecalis ETw23 se logró una mayor tasa de crecimiento entre las 6 y 10 h, y una mayor biomasa en MRScit. Después de 24 h de crecimiento E. faecium ETw7 alcanzó un nivel de 20,4 μM de diacetilo en MRS y 26,1 μM en MRScit, mientras que E. faecalis ETw23 logró niveles de 41,8 μM y 61,7 μM, respectivamente. Los resultados de este estudio sugieren que E. faecalis ETw23 puede contribuir en el desarrollo de aromas en quesos a través de su rol en la producción de diacetilo.
Olesen, M.; Gudmund-Høyer, E.; Norsker, Merete
: Seven healthy volunteers ingested in random order on seven different days: 20 g SPP; bread made of 180 g wheat flour served with 20 g raw SPP; bread baked of 180 g wheat flour and 20 g SPP; bread made from 180 g what flour; 20 g lactulose; 20 g oat bran; and 20 g wheat bran. The hydrogen breath test...... was used to evaluate oro-coecal transit time (OCTT) and fermentation. RESULTS: Fermentation of SPP yielded a measurable increase in end-expiratory H2. The total incremental increase in end expiratory H2 due to SPP was unaffected of whether SPP was served alone, as the raw flour served with bread, or baked...... into bread. The OCTT for raw SPP was significantly delayed compared to lactulose (P = 0.01). The OCTT for SPP baked into bread was significantly delayed compared to raw SPP (P = 0.01), indicating that SPP may be used as a marker of oro-coecal transit time for as well the fluid phase as the solid phase...
Dumani, Aysin; Guvenmez, Hatice Korkmaz; Yilmaz, Sehnaz; Yoldas, Oguz; Kurklu, Zeliha Gonca Bek
Aim. The purpose of this study was to compare the in vitro efficacy of calcium hypochlorite (Ca[OCl]2) and sodium hypochlorite (NaOCl) associated with sonic (Vibringe) irrigation system in root canals which were contaminated with Enterococcus faecalis. Material and Methods. The root canals of 84 single-rooted premolars were enlarged up to a file 40, autoclaved, inoculated with Enterococcus faecalis, and incubated for 21 days. The samples were divided into 7 groups according to the irrigation protocol: G0: no treatment; G1: distilled water; G2: 2.5% NaOCl; G3: 2.5% Ca(OCl)2; G4: distilled water with sonic activation; G5: 2.5% NaOCl with sonic activation; and G6: 2.5% Ca(OCl)2 with sonic activation. Before and after decontamination procedures microbiological samples were collected and the colony-forming units were counted and the percentages of reduction were calculated. Results. Distilled water with syringe irrigation and sonic activation groups demonstrated poor antibacterial effect on Enterococcus faecalis compared to other experimental groups (p irrigation systems with Ca(OCl)2 and NaOCl. Conclusion. The antimicrobial property of Ca(OCl)2 has been investigated and compared with that of NaOCl. Both conventional syringe irrigation and sonic irrigation were found effective at removing E. faecalis from the root canal of extracted human teeth. PMID:27218106
Full Text Available The general objective of this study was to examine the effect of bee product on the Enterococci colonization of chickens. Bee products were administered to both feed mixtures in various amounts in addition to the control group. First experimental group was with propolis in feed mixture with the addition of 200 mg propolis per 1 kg of compound and second group was with pollen with the addition of 250 mg pollen per 1 kg of compound. In this experiment, quantitative counts of Enterococci in ceca of 49-day-old chicken (Ross 308 using classical and MALDI TOF MS Biotyper method were investigated. Counts of Enterococci on Slanetz-Bartley agar were monitored. Enterococcus cells, isolated from gastrointestinal tract, were detected using MALDI TOF MS Biotyper. Counts of CFU of Enterococci were compared in experimental and control treatments, respectively. The lowest count was detected in the control experimental group. The highest count was detected in the first experimental group where was 200 mg of propolis added to 1 kg of feed mixture. Using MALDI TOF MS Biotyper, we identified the species range of the genera Enterococcus in the intestinal tract of broiler. Detected species from the genus Enterococcus were: E. avium, E. casseliflavus, E cecorum, E. faecalis, E. faecium, E. gallinarum, E. hirae and E. malodoratus. In the experimental groups (caecal samples were most frequent species of E. avium E. faecium and E. gallinarum.
Körpe, Didem Aksoy; İşerı, Özlem Darcansoy; Sahin, Feride Iffet; Cabi, Evren; Haberal, Mehmet
The aim of this study was to comparatively evaluate antibacterial activities of methanol (MetOH) and aqueous (dw) leaf (L), root (R) and seed (S) extracts of Urtica dioica L. (Ud; stinging nettle) and Urtica pilulifera L. (Up; Roman nettle) on both food- and plant-borne pathogens, with total phenolic contents and DPPH radical scavenging activities (DRSA). MetOH extracts of leaves and roots of U. dioica had the highest DRSA. Extracts with high antibacterial activity were in the order Up-LMetOH (13/16) > Ud-SMetOH (11/16) > Up-SMetOH (9/16). Results obtained with Up-SMetOH against food spoiling Bacillus pumilus, Shigella spp. and Enterococcus gallinarum with minimum inhibitory concentrations (MICs) in 128-1024 μg/ml range seem to be promising. Up-SMetOH also exerted strong inhibition against Clavibacter michiganensis with a considerably low MIC (32 μg/ml). Ud-SMetOH and Up-LMetOH were also effective against C. michiganensis (MIC = 256 and 1024 μg/ml, respectively). Ud-SMetOH and Ud-RMetOH had also antimicrobial activity against Xanthomonas vesicatoria (MIC = 512 and 1024 μg/ml, respectively). Results presented here demonstrate high-antibacterial activity of U. pilulifera extracts and U. dioica seed extract against phytopathogens for the first time, and provide the most comprehensive data on the antibacterial activity screening of U. pilulifera against food-borne pathogens. Considering limitations in plant disease control, antibacterial activities of these extracts would be of agricultural importance.
Bertsch, David; Muelli, Mirjam; Weller, Monika; Uruty, Anaïs; Lacroix, Christophe; Meile, Leo
The aims of this study were to assess antibiotic resistance pheno- and genotypes in foodborne, clinical, and environmental Listeria isolates, as well as to elucidate the horizontal gene transfer potential of detected resistance genes. A small fraction of in total 524 Listeria spp. isolates (3.1%) displayed acquired antibiotic resistance mainly to tetracycline (n = 11), but also to clindamycin (n = 4) and trimethoprim (n = 3), which was genotypically confirmed. In two cases, a tetracycline resistance phenotype was observed together with a trimethoprim resistance phenotype, namely in a clinical L. monocytogenes strain and in a foodborne L. innocua isolate. Depending on the applied guidelines, a differing number of isolates (n = 2 or n = 20) showed values for ampicillin that are on the edge between intermediate susceptibility and resistance. Transferability of the antibiotic resistance genes from the Listeria donors, elucidated in vitro by filter matings, was demonstrated for genes located on transposons of the Tn916 family and for an unknown clindamycin resistance determinant. Transfer rates of up to 10(-5) transconjugants per donor were obtained with a L. monocytogenes recipient and up to 10(-7) with an Enterococcus faecalis recipient, respectively. Although the prevalence of acquired antibiotic resistance in Listeria isolates from this study was rather low, the transferability of these resistances enables further spread in the future. This endorses the importance of surveillance of L. monocytogenes and other Listeria spp. in terms of antibiotic susceptibility. © 2014 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.
Labbe, Benjamin D; Kristich, Christopher J
Transmembrane Ser/Thr kinases containing extracellular PASTA domains are ubiquitous among Actinobacteria and Firmicutes Such PASTA kinases regulate critical processes, including antibiotic resistance, cell division, toxin production, and virulence, and are essential for viability in certain organisms. Based on in vitro studies with purified extracellular and intracellular fragments of PASTA kinases, a model for signaling has been proposed, in which the extracellular PASTA domains bind currently undefined ligands (typically thought to be peptidoglycan, or fragments thereof) to drive kinase dimerization, which leads to enhanced kinase autophosphorylation and enhanced phosphorylation of substrates. However, this model has not been rigorously tested in vivo Enterococcus faecalis is a Gram-positive intestinal commensal and major antibiotic-resistant opportunistic pathogen. In E. faecalis , the PASTA kinase IreK drives intrinsic resistance to cell wall-active antimicrobials, suggesting that such antimicrobials may trigger IreK signaling. Here we show that IreK responds to cell wall stress in vivo by enhancing its phosphorylation and that of a downstream substrate. This response requires both the extracellular PASTA domains and specific phosphorylatable residues in the kinase domain. Thus, our results provide in vivo evidence, with an intact full-length PASTA kinase in its native physiological environment, that supports the prevailing model of PASTA kinase signaling. In addition, we show that IreK responds to a signal associated with growth and/or cell division, in the absence of cell wall-active antimicrobials. Surprisingly, the ability of IreK to respond to growth and/or division does not require the extracellular PASTA domains, suggesting that IreK monitors multiple parameters for sensory input in vivo IMPORTANCE Transmembrane Ser/Thr kinases containing extracellular PASTA domains are ubiquitous among Actinobacteria and Firmicutes and regulate critical processes. The
Honsa, Erin S; Cooper, Vaughn S; Mhaissen, Mohammed N; Frank, Matthew; Shaker, Jessica; Iverson, Amy; Rubnitz, Jeffrey; Hayden, Randall T; Lee, Richard E; Rock, Charles O; Tuomanen, Elaine I; Wolf, Joshua; Rosch, Jason W
Serious bacterial infections in immunocompromised patients require highly effective antibacterial therapy for cure, and thus, this setting may reveal novel mechanisms by which bacteria circumvent antibiotics in the absence of immune pressure. Here, an infant with leukemia developed vancomycin-resistant Enterococcus faecium (VRE) bacteremia that persisted for 26 days despite appropriate antibiotic therapy. Sequencing of 22 consecutive VRE isolates identified the emergence of a single missense mutation (L152F) in relA, which constitutively activated the stringent response, resulting in elevated baseline levels of the alarmone guanosine tetraphosphate (ppGpp). Although the mutant remained susceptible to both linezolid and daptomycin in clinical MIC testing and during planktonic growth, it demonstrated tolerance to high doses of both antibiotics when growing in a biofilm. This biofilm-specific gain in resistance was reflected in the broad shift in transcript levels caused by the mutation. Only an experimental biofilm-targeting ClpP-activating antibiotic was able to kill the mutant strain in an established biofilm. The relA mutation was associated with a fitness trade-off, forming smaller and less-well-populated biofilms on biological surfaces. We conclude that clinically relevant relA mutations can emerge during prolonged VRE infection, causing baseline activation of the stringent response, subsequent antibiotic tolerance, and delayed eradication in an immunocompromised state. The increasing prevalence of antibiotic-resistant bacterial pathogens is a major challenge currently facing the medical community. Such pathogens are of particular importance in immunocompromised patients as these individuals may favor emergence of novel resistance determinants due to lack of innate immune defenses and intensive antibiotic exposure. During the course of chemotherapy, a patient developed prolonged bacteremia with vancomycin-resistant Enterococcus faecium that failed to clear
Gao, Lujuan; Sun, Yi; He, Chengyan; Li, Ming; Zeng, Tongxiang; Lu, Qiaoyun
Infections of Exophiala spp. and Fusarium spp. are often chronic and recalcitrant. Systemic disseminations, which mostly occur in immunocompromised patients, are often refractory to available antifungal therapies. The conserved target of rapamycin (TOR) orchestrates cell growth and proliferation in response to nutrients and growth factors, which are important for pathogenicity and virulence. INK128 is a second-generation ATP-competitive TOR inhibitor, which binds the TOR catalytic domain and selectively inhibits TOR. In the present study, we investigated the in vitro activities of INK128 alone and the interactions of INK128 with conventional antifungal drugs including itraconazole, voriconazole, posaconazole, and amphotericin B against 18 strains of Exophiala spp. and 10 strains of Fusarium spp. via broth microdilution checkerboard technique system adapted from Clinical and Laboratory Standards Institute broth microdilution method M38-A2. INK128 alone was inactive against all isolates tested. However, favorable synergistic effects between INK128 and voriconazole were observed in 61% Exophiala strains and 60% Fusarium strains, despite Fusarium strains exhibited high MIC values (4-8 μg/ml) against voriconazole. In addition, synergistic effects of INK128/itraconazole were shown in 33% Exophiala strains and 30% Fusarium strains, while synergy of INK128/posaconazole were observed in 28% Exophiala strains and 30% Fusarium strains. The effective working ranges of INK128 were 0.125-2 μg/ml and 1-4 μg/ml against Exophiala isolates and Fusarium isolates, respectively. No synergistic effect was observed when INK128 was combined with amphotericin B. No antagonism was observed in all combinations. In conclusion, INK128 could enhance the in vitro antifungal activity of voriconazole, itraconazole and posaconazole against Exophiala spp. and Fusarium spp., suggesting that azoles, especially voriconazole, combined with TOR kinase inhibitor might provide a potential strategy to
Fu, B; Jiang, Q; Liu, H-B; Liu, H
The presence of viable but nonculturable (VBNC) bacterial pathogens which often fail to be detected by cultivation and can regain the cultivability if the living conditions improve were reported. The objective of this study was to determine the occurrence of VBNC Salmonella spp. and Shigella spp. in the biosolids during anaerobic digestion and its reactivation during the cake storage. The occurrence of VBNC Salmonella spp. and Shigella spp. during mesophilic, temperature-phased, thermophilic anaerobic digestion of sewage sludge and the subsequent storage were studied by RT-qPCR and most probable number (MPN) method. The VBNC incidence of Salmonella spp. and Shigella spp. during thermophilic digestion was four orders of magnitude higher than those of mesophilic digestion. Accordingly, higher resuscitation ratio of VBNC pathogens was also achieved in thermophilic digested sludge. As a result, the culturable Salmonella typhimurium contents in thermophilic digested sludge after cake storage were two orders of magnitude higher than mesophilic digestion. Both quantitative PCR and reverse transcription quantitative PCR assay results showed the two bacterial counting numbers remained stable throughout the cake storage. The results indicate that the increase in the culturable Salmonella spp. and Shigella spp. after centrifugal dewatering was attributed to the resuscitation from the VBNC state to the culturable state. Thermophilic anaerobic digestion mainly induced Salmonella spp. and Shigella spp. into VBNC state rather than killed them, suggesting that the biological safety of sewage sludge by temperature-phased anaerobic digestion should be carefully assessed. © 2015 The Society for Applied Microbiology.
Paola Mendes Milanesi
Full Text Available This study aimed i to quantify the occurrence of Fusarium spp. and Trichoderma spp. in rhizospheric soil, with and without symptoms of Sudden Death Syndrome (SDS in eight soybean genotypes; ii morphologically identify isolates of Fusarium spp. from roots with SDS; iii evaluate the antagonism between Trichoderma spp. and Fusarium spp. isolates from rhizospheric soil and roots from with and without SDS, respectively; and iv characterize through the ITS1-5.8S-ITS2 region of rDNA the isolates of Trichoderma spp. with better performance in the direct confrontation. The sampling of soil and roots was performed in an experimental area located in Cruz Alta, RS, Brazil. In the laboratory, serial dilutions of soil samples, counting of the number of Colony Forming Units (UFCs/g-1 of rhizospheric soil were performed as well as isolation for identification of isolates of Fusarium spp. and Trichoderma spp. and testing of direct confrontation. There were significant differences between the population of Trichoderma spp. in the rhizosphere of plants with and without symptoms of SDS. For the population of Fusarium spp., significant difference was observed only in the rhizosphere of plants without symptoms of SDS. In diseased roots the following species were identified: F. solani, F. avenaceum, F. graminearum, F. oxysporum and F. verticillioides. In the test of direct confrontation, eight isolates of Trichoderma spp. achieved the best performance in the antagonism to Fusarium spp. and Trichoderma spp. from areas with symptoms of SDS had a higher control efficiency in vitro. These isolates showed high similarity to the species of T. koningii agregate.
Mayer, Fabiana Quoos; Dos Reis, Emily Marques; Bezerra, André Vinícius Andrade; Cerva, Cristine; Rosa, Júlio; Cibulski, Samuel Paulo; Lima, Francisco Esmaile Sales; Pacheco, Susi Missel; Rodrigues, Rogério Oliveira
The present study aimed to investigate the frequency of pathogenic Leptospira spp. in Brazilian bats and to determine possible risk factors associated to it. Ninety two bats of 12 species were evaluated. Whole genomic DNA from kidneys was extracted and real-time PCR specific to pathogenic Leptospira spp. was applied. Association between the frequency of specimens positive for Leptospira spp. and sex, age, bat species or family, season of collection, geographic localization and feeding habits was evaluated. The results showed that 39.13% of analyzed bats were found positive for Leptospira spp. Nine bat species had at least one positive result. There was no association among the evaluated variables and frequency of pathogenic Leptospira spp. Although the limitations due to lack of Leptospira spp. isolation, leptospiral carriage was demonstrated in bats of different species from southern Brazil, which reinforces the need for surveillance of infectious agents in wild animals. Copyright © 2017 Elsevier Ltd. All rights reserved.
Trimoulinard, A; Beral, M; Henry, I; Atiana, L; Porphyre, V; Tessier, C; Leclercq, A; Cardinale, E
One of the most popular meat products of the local "cuisine" is sausage composed with 100% chicken or 100% pork. In this study, we aimed to determine the presence of Salmonella spp., Campylobacter spp. and Listeria spp. in chicken- and pork-sausages, quantify Salmonella spp. population and identify the factors that could be associated with contamination in the outlets. Two hundred and three batches of pork and chicken sausages were randomly collected from 67 local outlets (supermarkets, groceries and butcher shops). Salmonella spp. was detected in 11.8% (95% confidence interval (CI): [10.0; 13.5]) of samples, Campylobacter spp. in 1.5% [0.7; 4.2] and Listeria monocytogenes in 5.9% [4.4; 7.3]. Most probable number of Salmonella spp. varied between 6cfu per gram to 320cfu per gram. Salmonella serotypes isolated from pork and chicken sausages were S. Typhimurium (45.8%), S. London (20.8%), S. Derby (16.7%), S. Newport (8.33%), S. Blockley (4.2%) and S. Weltevreden (4.17%). Using a logistic (mixed-effect) regression model, we found that Salmonella spp. contamination was positively associated with sausages sold in papers or plastic bags and no control of rodents. Chicken sausages were associated with a decreasing risk of Salmonella contamination. Listeria monocytogenes contamination was positively associated with the presence of fresh rodent droppings in the outlet and negatively when the staff was cleaning regularly their hands with soap and water or water only. All the sampled outlets of Reunion Island were not equivalent in terms of food safety measures. Increasing awareness of these traders remains a cornerstone to limit the presence of Salmonella spp. and Listeria spp. in sausages, particularly in a tropical context (high temperature and humidity). Copyright © 2017 Elsevier B.V. All rights reserved.
Choi, Eun-Ju; Iwasa, Masahiro; Han, Kwon-Il; Kim, Wan-Jae; Tang, Yujiao; Hwang, Young Joung; Chae, Jeong Ryong; Han, Weon Cheol; Shin, Yu-Su; Kim, Eun-Kyung
Recent reports have shown the immunomodulatory effect of heat-killed lactic acid bacteria. Atopic dermatitis (AD) is an allergic skin disease, caused by immune dysregulation among other factors. The aim of this study was to assess the effect of heat-killed Enterococcus faecalis EF-2001 (EF-2001) on AD. We established an in vivo AD model by repeated local exposure of Dermatophagoides farinae extract (DFE; house dust mite extract) and 2,4-dinitrochlorobenzene (DNCB) to the ears of mice. After oral administration of EF-2001 for four weeks, the epidermal and dermal ear thickness, mast cell infiltration, and serum immunoglobulin levels were measured. In addition, the gene expression levels of pathogenic cytokines in the ears, lymph nodes, and splenocytes were assayed. EF-2001 attenuated AD symptoms based on the ear thickness, histopathological analysis, and serum immunoglobulin levels. Moreover, EF-2001 decreased the DFE/DNCB-induced expression of various pathogenic cytokines in the ears, lymph nodes, and splenocytes. These results suggest that EF-2001 has therapeutic potential in the treatment of AD owing to its immunomodulatory effects. PMID:26959058
Guiton, Pascale S.; Barnes, Aaron M. T.; Manias, Dawn A.; Chuang-Smith, Olivia N.; Kohler, Petra L.; Spaulding, Adam R.; Hultgren, Scott J.; Schlievert, Patrick M.; Dunny, Gary M.
Enterococcus faecalis is part of the human intestinal microbiome and is a prominent cause of health care-associated infections. The pathogenesis of many E. faecalis infections, including endocarditis and catheter-associated urinary tract infection (CAUTI), is related to the ability of clinical isolates to form biofilms. To identify chromosomal genetic determinants responsible for E. faecalis biofilm-mediated infection, we used a rabbit model of endocarditis to test strains with transposon insertions or in-frame deletions in biofilm-associated loci: ahrC, argR, atlA, opuBC, pyrC, recN, and sepF. Only the ahrC mutant was significantly attenuated in endocarditis. We demonstrate that the transcriptional regulator AhrC and the protease Eep, which we showed previously to be an endocarditis virulence factor, are also required for full virulence in murine CAUTI. Therefore, AhrC and Eep can be classified as enterococcal biofilm-associated virulence factors. Loss of ahrC caused defects in early attachment and accumulation of biofilm biomass. Characterization of ahrC transcription revealed that the temporal expression of this locus observed in wild-type cells promotes initiation of early biofilm formation and the establishment of endocarditis. This is the first report of AhrC serving as a virulence factor in any bacterial species. PMID:23460519
Full Text Available Enterococcus lactis IW5 was obtained from human gut and the potential probiotic characteristics of this organism were then evaluated. Results showed that this strain was highly resistant to low pH and high bile salt and adhered strongly to Caco-2 human epithelial colorectal cell lines. The supernatant of E. lactis IW5 strongly inhibited the growth of several pathogenic bacteria and decreased the viability of different cancer cells, such as HeLa, AGS, HT-29, and MCF-7. Conversely, E. lactis IW5 did not inhibit the viability of normal FHs-74 cells. This strain did not generate toxic enzymes, including β-glucosidase, β-glucuronidase, and N-acetyl-β-glucosaminidase and was highly susceptible to ampicillin, gentamycin, penicillin, vancomycin, clindamycin, sulfamethoxazol, and chloramphenicol but resistant to erythromycin and tetracyclin. This study provided evidence for the effect of E. lactis IW5 on cancer cells. Therefore, E. lactis IW5, as a bioactive therapeutics, should be subjected to other relevant tests to verify the therapeutic suitability of this strain for clinical applications.
Full Text Available Recent reports have shown the immunomodulatory effect of heat-killed lactic acid bacteria. Atopic dermatitis (AD is an allergic skin disease, caused by immune dysregulation among other factors. The aim of this study was to assess the effect of heat-killed Enterococcus faecalis EF-2001 (EF-2001 on AD. We established an in vivo AD model by repeated local exposure of Dermatophagoides farinae extract (DFE; house dust mite extract and 2,4-dinitrochlorobenzene (DNCB to the ears of mice. After oral administration of EF-2001 for four weeks, the epidermal and dermal ear thickness, mast cell infiltration, and serum immunoglobulin levels were measured. In addition, the gene expression levels of pathogenic cytokines in the ears, lymph nodes, and splenocytes were assayed. EF-2001 attenuated AD symptoms based on the ear thickness, histopathological analysis, and serum immunoglobulin levels. Moreover, EF-2001 decreased the DFE/DNCB-induced expression of various pathogenic cytokines in the ears, lymph nodes, and splenocytes. These results suggest that EF-2001 has therapeutic potential in the treatment of AD owing to its immunomodulatory effects.
Full Text Available The objective of this study was to investigate the occurrence of vancomycin-resistant Enterococcus (VRE cross-transmission between two patient groups (long-term dialysis and kidney transplant patients. Molecular typing, by automated ribotyping with the RiboPrinter Microbial Characterization System (Qualicon, USA, was used to analyze VRE isolates from 31 fecal samples of 320 dialysis patients and 38 fecal samples of 280 kidney transplant patients. Clonal spread of E. faecalis and E. casseliflavus was observed intragroup, but not between the two groups of patients. In turn, transmission of E. gallinarum and E. faecium between the groups was suggested by the finding of vancomycin-resistant isolates belonging to the same ribogroup in both dialysis and transplant patients. The fact that these patients were colonized by VRE from the same ribogroup in the same health care facility provides evidence for cross-transmission and supports the adoption of stringent infection control measures to prevent dissemination of these bacteria.
de Sousa, Marylane; Manzo, Ricardo M; García, José L; Mammarella, Enrique J; Gonçalves, Luciana R B; Pessela, Benevides C
l-Arabinose isomerase (EC 188.8.131.52) (l-AI) from Enterococcus faecium DBFIQ E36 was overproduced in Escherichia coli by designing a codon-optimized synthetic araA gene. Using this optimized gene, two N- and C-terminal His-tagged-l-AI proteins were produced. The cloning of the two chimeric genes into regulated expression vectors resulted in the production of high amounts of recombinant N -His-l-AI and C -His-l-AI in soluble and active forms. Both His-tagged enzymes were purified in a single step through metal-affinity chromatography and showed different kinetic and structural characteristics. Analytical ultracentrifugation revealed that C -His-l-AI was preferentially hexameric in solution, whereas N -His-l-AI was mainly monomeric. The specific activity of the N -His-l-AI at acidic pH was higher than that of C -His-l-AI and showed a maximum bioconversion yield of 26% at 50 °C for d-tagatose biosynthesis, with Km and Vmax parameters of 252 mM and 0.092 U mg -1 , respectively. However, C -His-l-AI was more active and stable at alkaline pH than N -His-l-AI. N -His-l-AI follows a Michaelis-Menten kinetic, whereas C -His-l-AI fitted to a sigmoidal saturation curve.
Full Text Available Enterococcus faecalis is considered a pathogen responsible for hospital infections and, due to its frequent multi-resistant profile, has caused preoccupations among many medical authorities. The objective of this study was to determine the antimicrobial susceptibility of 74 strains isolated from blood cultures and purulent secretions to vancomycin and ciprofloxacin through the Minimum Inhibitory Concentration (MIC and Minimum Bactericidal Concentration (MBC by using the Microdilution test. The results showed a greater efficacy of vancomycin compared to ciprofloxacin (98.6% of the strains were inhibited by vancomycin at lower concentrations: 0.06 - 1 µg/ml. However, in the MBC analysis 73% of the strains showed a MBC of vancomycin only at high concentrations (equal to or higher than 64 µg/ml. For ciprofloxacin, the strains showed a broad sensitivity with MICs and MBCs distributed along all the MIC classes. Results also revealed a probability that some strains are tolerant to vancomycin, which indicates the need of other tests to confirm this characteristic.
Full Text Available Objectives: The aim of the study was to compare manual dynamic agitation with sonic agitation on removal of intra radicular Enterococcus faecalis (E. faecalis biofilm. Material and Methods: Extracted mandibular premolars for orthodontic purpose were sectioned at cervical level and divided into three groups (n = 30. The root canals were instrumented using Protaper rotary instruments up to apical file F4. Roots were sterilized and E. faecalis bacteria were incubated within their root canal space for four weeks. Confirmation of biofilm was done using scanning electron microscopy and Gram staining. All groups were irrigated with side vented needle by using three percent sodium hypochlorite (NaOCl for 60 seconds. Two experimental groups were agitated with manual dynamic agitation (with master gutta-percha cone and sonic agitation (EndoActivator. Remaining bacteria were collected using sterile paper point, which were incubated inside brain-heart infusion (BHI broth to check turbidity. The turbid broth was streaked on blood agar plate for colony counts. Result: Both experimental groups showed highly significant difference in their mean colony count when compared with control group; with P < 0.001. Conclusion: Passive sonic agitation with EndoActivator has proven to be the best irrigating system followed by manual dynamic agitation and conventional needle irrigation.
Luisa Marcela Villamil Díaz
Full Text Available 120 bacteria were isolated from tilapia intestine and screened according to the antibacterial activity against pathogens such as Aeromonas hydrophila, Edwardsiella tarda and Streptococcus agalactiae, the ability to adhere to intestinal mucus and growth kinetics. The selected bacteria were identified by 16S rRNA sequencing of and were identified as Exigobacterium sp. I9, Enterococcus faecalis I15 and Myroides odoratimimus I19. Furthermore, the in vivo effect on fish growth was assessed by the addition of selected bacteria to juvenile Oreochromis niloticus feed (106 CFU / g, for 15 days. Survival was also determined after a challenge by intraperitoneal injection of E. tarda (100 μL of 105 UFC / mL. The three selected bacteria increased the specific growth rate, reduced mortality of fish during the experimental challenge with E. tarda and did not cause mortality during the addition in the feed. The positive effects observed in vivo are possibly associated with in vitro activity; however, for biosafety reasons, it is recommended that further studies of Exigobacterium sp. I9 and E. faecalis I15 may be carried out since this genus have been reported of as causative agents of fish mortality, whereas in the case of M. odoratimimus I19, verification of positive effects at a higher production scales is desirable.
Full Text Available Background: Enterococcus faecalis (E. faecalis is a gram positive oral pathogen that reported at the main agent infection of endodontic treatment. Its activities are influenced by the virulence factors facilitating the interaction process between agents with host cells. Like aggregation substance, cytolysin, extracellular superoxide, gelatinase, hyaluronidase, sex pheromones, and surface adhesions molecules. Plant extracts are reported as the material antibacterial as well as E. faecalis in pathogenesis of endodontic infections. Purpose: Purpose of this study was to analyse of sarang semut extracts (Myrmecodia Pendens Merr. & Perry towards sensitivity of E. faecalis. Method: This research used the methanol extract of sarang semut, E. faecalis ATCC 29212, and fosfomycin also chlorhexidine as the positive controls. Whereas, Bradford protein method was measured the concentration of the surface protein of E. faecalis and active component of the sarang semut extract. Result: Generally, the sarang semut extract possessed low sensitivity toward E. faecalis (≤ 13 mm, but on the concentrations of 100 µg/ml and 75 µg/ml better than inhibition of other concentrations, round 10.6-11.6 (mm. Specifically, on 100 µg/ml has indicator the minimal bactericidal concentration (MBC on E. faecalis. Whereas minimal inhibition concentration (MIC on the concentration of 3,125 µg/ml. Conclusion: Based on MBC and MIC assay, the extract of sarang semut has potential effects to adherence growth of E. faecalis, mainly on the highest concentration 100 µg/ml also MIC on 3,125 µg/ ml.
Vatkar, Niranjan Ashok; Hegde, Vivek; Sathe, Sucheta
To compare the vitality of Enterococcus faecalis within dentinal tubules after subjected to five root canal disinfection methods. Dentin blocks (n = 60) were colonized with E. faecalis. After 4 weeks of incubation, the dentin blocks were divided into one control and five test groups (n = 10 each). The root canals of test groups were subjected to one of the disinfection methods, namely, normal saline (NS), sodium hypochlorite (NaOCl), chlorhexidine digluconate (CHX), neodymium-doped yttrium aluminum garnet (Nd: YAG) laser, and diode laser. The effect of disinfection methods was assessed by LIVE/DEAD BacLight stain under the confocal laser scanning microscopy to determine the "zone of dead bacteria" (ZDB). Mean values were calculated for ZDB and the difference between groups was established. Penetration of E. faecalis was seen to a depth of >1000 μm. Viable bacteria were detected with NS irrigation. NaOCl and CHX showed partial ZDB. When the root canals were disinfected with Nd: YAG and diode lasers, no viable bacteria were found. E. faecalis has the ability to colonize inside dentinal tubules to a depth of >1000 μm. In contrast to conventional irrigants, both Nd: YAG and diode lasers were effective in eliminating the vitality of E. faecalis. NS, NaOCl, and CHX showed viable bacteria remaining in dentinal tubules.
Full Text Available The species Enterococcus faecalis is able to catabolise the amino acid tyrosine into the biogenic amine tyramine by the tyrosine decarboxilase (TDC pathway Ladero et al. (2012 . The TDC cluster comprises four genes: tyrS, an aminoacyl-tRNA synthetase-like gene; tdcA, which encodes the tyrosine decarboxylase; tyrP, a tyrosine/tyramine exchanger gene and nhaC-2, which encodes an Na+/H+ antiporter and whose role in the tyramine biosynthesis remains unknown . In E. faecalis V583 the last three genes are co-transcribed as a single polycistronic mRNA forming the catabolic operon, while tyrS is transcribed independently of the catabolic genes as a monocistronic mRNA . The catabolic operon is transcriptionally induced by tyrosine and acidic pH. On the opposite, the tyrS expression is repressed by tyrosine concentrations . In this work we report the transcriptional profiling of the TDC cluster deletion mutant (E. faecalis V583 ΔTDC  compared to the wild-type strain, both grown in M17 medium supplemented with tyrosine. The transcriptional profile data of TDC cluster-regulated genes were deposited in the Gene Expression Omnibus (GEO database under accession no. GSE77864.
Miller, Corwin; Kong, Jiayi; Tran, Truc T; Arias, Cesar A; Saxer, Gerda; Shamoo, Yousif
With increasing numbers of hospital-acquired antibiotic resistant infections each year and staggering health care costs, there is a clear need for new antimicrobial agents, as well as novel strategies to extend their clinical efficacy. While genomic studies have provided a wealth of information about the alleles associated with adaptation to antibiotics, they do not provide essential information about the relative importance of genomic changes, their order of appearance, or potential epistatic relationships between adaptive changes. Here we used quantitative experimental evolution of a single polymorphic population in continuous culture with whole-genome sequencing and allelic frequency measurements to study daptomycin (DAP) resistance in the vancomycin-resistant clinical pathogen Enterococcus faecalis S613. Importantly, we sustained both planktonic and nonplanktonic (i.e., biofilm) populations in coculture as the concentration of antibiotic was raised, facilitating the development of more ecological complexity than is typically observed in laboratory evolution. Quantitative experimental evolution revealed a clear order and hierarchy of genetic changes leading to resistance, the signaling and metabolic pathways responsible, and the relative importance of these mutations to the evolution of DAP resistance. Despite the relative simplicity of this ex vivo approach compared to the ecological complexity of the human body, we showed that experimental evolution allows for rapid identification of clinically relevant adaptive molecular pathways and new targets for drug design in pathogens.
Cao Yingguang; Yang Ping; Lu Xinpei; Xiong Zilan; Ye Tao; Xiong Qing; Sun Ziyong
Enterococcus faecalis (E. faecalis) is a microorganism that can survive extreme challenges in obturated root canals. The aim of this study was to evaluate the efficacy of a non-thermal atmospheric pressure plasma plume against E. faecalis in vitro. A non-thermal atmospheric pressure plasma jet device which could generate a cold plasma plume carrying a peak current of 300 mA was used. The antibacterial efficacy of this device against E. faecalis and its biofilm under different conditions was detected. The antibacterial efficacy of the plasma against E. faecalis and Staphylococcus aureus (S. aureus) was also evaluated. After plasma treatment, the average diameter of inhibition zone on S. aureus and E. faecalis was 2.62±0.26 cm and 1.06±0.30 cm, respectively (P < 0.05). The diameter was increased with prolongation of the treatment duration. The diameters of inhibition zone of the sealed Petri dishes were larger than those of the uncovered Petri dishes. There was significant difference in colony-forming units between plasma group and control group on E. faecalis biofilm (P < 0.01). The transmission electron microscopy revealed that the ultrastructural changes cytoderm of E. faecalis were observed after treatment for 2 min. It is concluded that the non-thermal atmospheric pressure plasma could serve as an effective adjunct to standard endodontic microbial treatment.
Full Text Available Enterococcus faecalis is an important opportunistic pathogen which is frequently detected in mineral water and spring water for human consumption and causes human urinary tract infections, endocarditis and neonatal sepsis. The aim of this study was to determine the prevalence, virulence genes, antimicrobial resistance and genetic diversity of E. faecalis from mineral water and spring water in China. Of 314 water samples collected from January 2013 to January 2014, 48 samples (15.3% were contaminated E. faecalis. The highest contamination rate occurred in activated carbon filtered water of spring water (34.5%, followed by source water of spring water (32.3% and source water of mineral water (6.4%. The virulence gene test of 58 E. faecalis isolates showed that the detection rates of asa1, ace, cylA, gelE and hyl were 79.3, 39.7, 0, 100, 0%, respectively. All 58 E. faecalis isolates were not resistant to 12 kinds of antibiotics (penicillin, ampicillin, linezolid, quinupristin/dalfopristin, vancomycin, gentamicin, streptomycin, ciprofloxacin, levofloxacin, norfloxacin, nitrofurantoin, and tetracycline. Enterobacterial repetitive intergenic consensus-PCR classified 58 isolates and three reference strains into nine clusters with a similarity of 75%. This study is the first to investigate the prevalence of E. faecalis in mineral water and spring water in China. The results of this study suggested that spring water could be potential vehicles for transmission of E. faecalis.
Marylane de Sousa
Full Text Available l-Arabinose isomerase (EC 184.108.40.206 (l-AI from Enterococcus faecium DBFIQ E36 was overproduced in Escherichia coli by designing a codon-optimized synthetic araA gene. Using this optimized gene, two N- and C-terminal His-tagged-l-AI proteins were produced. The cloning of the two chimeric genes into regulated expression vectors resulted in the production of high amounts of recombinant N-His-l-AI and C-His-l-AI in soluble and active forms. Both His-tagged enzymes were purified in a single step through metal-affinity chromatography and showed different kinetic and structural characteristics. Analytical ultracentrifugation revealed that C-His-l-AI was preferentially hexameric in solution, whereas N-His-l-AI was mainly monomeric. The specific activity of the N-His-l-AI at acidic pH was higher than that of C-His-l-AI and showed a maximum bioconversion yield of 26% at 50 °C for d-tagatose biosynthesis, with Km and Vmax parameters of 252 mM and 0.092 U mg−1, respectively. However, C-His-l-AI was more active and stable at alkaline pH than N-His-l-AI. N-His-l-AI follows a Michaelis-Menten kinetic, whereas C-His-l-AI fitted to a sigmoidal saturation curve.
Full Text Available The robust physiology of Enterococcus faecalis facilitates tolerance to various stresses. We here report the transcriptional response of E. faecalis V583 to growth in the presence of 6.5% NaCl. Among the early responses observed was an immediate down-regulation of mscL, accompanied by an up-regulation of genes predicted to be involved in uptake of extracellular potassium and glycine betaine. The high NaCl concentration also induced expression of chaperons and cell envelope related traits, such as the enterococcal polysaccharide antigen (epa locus. Functional genetic analysis revealed reduced salt stress resistance in both epaB and epaE mutants. The reduced salt resistance phenotype associated with the epaB mutant was restored by complementation, hence demonstrating a role of Epa in the physiological robustness of E. faecalis. Furthermore, we demonstrate that Epa confers increased resistance towards multiple cell envelope stress-inducing factors. Accordingly, these findings delineate a potential link between the robust nature of E. faecalis and its ability to perform as a human pathogen, and provide a new perspective on the mechanisms by which Epa contributes to virulence. Notably, the high NaCl concentration also resulted in strict repression of the gelE-sprE operon and impaired gelatinase activity. We demonstrate that NaCl antagonize the GBAP-pheromone dependent induction in a concentration dependent manner.
Full Text Available Use of bacteriocins in food preservation has received great attention in recent years. The goal of this study is to characterize enterocin RM6 from Enterococcus faecalis OSY-RM6 and investigate its efficacy against Listeria monocytogenes in cottage cheese. Enterocin RM6 was purified from E. faecalis culture supernatant using ion exchange column, multiple C18-silica cartridges, followed by reverse-phase high-performance liquid chromatography. The molecular weight of enterocin RM6 is 7145.0823 as determined by mass spectrometry (MS. Tandem mass spectrometry (MS/MS analysis revealed that enterocin RM6 is a 70-residue cyclic peptide with a head-to-tail linkage between methionine and tryptophan residues. The peptide sequence of enterocin RM6 was further confirmed by sequencing the structural gene of the peptide. Enterocin RM6 is active against Gram-positive bacteria, including L. monocytogenes, Bacillus cereus, and methicillin-resistant Staphylococcus aureus (MRSA. Enterocin RM6 (final concentration in cottage cheese, 80 AU/mL caused a 4-log reduction in population of L. monocytogenes inoculated in cottage cheese within 30 min of treatment. Therefore, enterocin RM6 has potential applications as a potent antimicrobial peptide against foodborne pathogens in food.
Huang, En; Zhang, Liwen; Chung, Yoon-Kyung; Zheng, Zuoxing; Yousef, Ahmed E
Use of bacteriocins in food preservation has received great attention in recent years. The goal of this study is to characterize enterocin RM6 from Enterococcus faecalis OSY-RM6 and investigate its efficacy against Listeria monocytogenes in cottage cheese. Enterocin RM6 was purified from E. faecalis culture supernatant using ion exchange column, multiple C18-silica cartridges, followed by reverse-phase high-performance liquid chromatography. The molecular weight of enterocin RM6 is 7145.0823 as determined by mass spectrometry (MS). Tandem mass spectrometry (MS/MS) analysis revealed that enterocin RM6 is a 70-residue cyclic peptide with a head-to-tail linkage between methionine and tryptophan residues. The peptide sequence of enterocin RM6 was further confirmed by sequencing the structural gene of the peptide. Enterocin RM6 is active against Gram-positive bacteria, including L. monocytogenes, Bacillus cereus, and methicillin-resistant Staphylococcus aureus (MRSA). Enterocin RM6 (final concentration in cottage cheese, 80 AU/mL) caused a 4-log reduction in population of L. monocytogenes inoculated in cottage cheese within 30 min of treatment. Therefore, enterocin RM6 has potential applications as a potent antimicrobial peptide against foodborne pathogens in food.
Gupta, H; Malik, R K; Bhardwaj, A; Kaur, G; De, S; Kaushik, J K
Enterococcus faecium FH 99 was isolated from human faeces and selected because of its broad spectrum of inhibitory activity against several Gram-positive foodborne spoilage and pathogenic bacteria. Ent. faecium FH 99 accumulates enterocin in large number in early stationary phase of the growth. The enterocin FH 99 was stable over a wide pH range (2-10) and recovered activity even after treatment at high temperatures (10 min at 100°C). The enterocin was subjected to different purification techniques viz., gel filteration, cation exchange chromatography and reverse-phase high-performance liquid chromatography. The activity was eluted as one individual active fraction. SDSPAGE revealed a molecular weight of less than 6.5 kDa. Studies carried out to identify the genetic determinants for bacteriocin production showed that this trait may be plasmid encoded as loss in both of the plasmids (size>chromosomal DNA) led to loss in bacteriocin production by Ent. faecium FH 99. Ent. faecium strain FH 99 is a newly discovered high bacteriocin producer with Activity Units 1.8 × 10(5) AU ml(-1) and its characteristics indicate that it may have strong potential for application as a protective agent against pathogens and spoilage bacteria in foods.
Arbulu, Sara; Lohans, Christopher T; van Belkum, Marco J; Cintas, Luis M; Herranz, Carmen; Vederas, John C; Hernández, Pablo E
The solution structure of enterocin HF (EntHF), a class IIa bacteriocin of 43 amino acids produced by Enterococcus faecium M3K31, was evaluated by CD and NMR spectroscopy. Purified EntHF was unstructured in water, but CD analysis supports that EntHF adopts an α-helical conformation when exposed to increasing concentrations of trifluoroethanol. Furthermore, NMR spectroscopy indicates that this bacteriocin adopts an antiparallel β-sheet structure in the N-terminal region (residues 1-17), followed by a well-defined central α-helix (residues 19-30) and a more disordered C-terminal end (residues 31-43). EntHF could be structurally organized into three flexible regions that might act in a coordinated manner. This is in agreement with the absence of long-range nuclear Overhauser effect signals between the β-sheet domain and the C-terminal end of the bacteriocin. The 3D structure recorded for EntHF fits emerging facts regarding target recognition and mode of action of class IIa bacteriocins.
ABSTRACT Enterococci are ancient commensal bacteria that recently emerged as leading causes of antibiotic-resistant, hospital-acquired infection. Vancomycin-resistant enterococci (VRE) epitomize why drug-resistant enterococcal infections are a problem: VRE readily colonize the antibiotic-perturbed gastrointestinal (GI) tract where they amplify to large numbers, and from there, they infect other body sites, including the bloodstream, urinary tract, and surgical wounds. VRE are resistant to many antimicrobials and host defenses, which facilitates establishment at the site of infection and confounds therapeutic clearance. Having evolved to colonize the GI tract, VRE are comparatively ill adapted to the human bloodstream. A recent study by Honsa and colleagues (E. S. Honsa et al., mBio 8:e02124-16, 2017, https://doi.org/10.1128/mBio.02124-16) found that a strain of vancomycin-resistant Enterococcus faecium evolved antibiotic tolerance within the bloodstream of an immunocompromised host by activating the stringent response through mutation of relA. Precisely how VRE colonize and infect and the selective pressures that led to the outgrowth of relA mutants are the subjects of ongoing research. PMID:28223450
Linares, Daniel M; Perez, Marta; Ladero, Victor; Del Rio, Beatriz; Redruello, Begoña; Martin, M Cruz; Fernandez, María; Alvarez, Miguel A
Scientific interest in Enterococcus faecalis has increased greatly over recent decades. Some strains are involved in food fermentation and offer health benefits, whereas others are vancomycin-resistant and cause infections that are difficult to treat. The limited availability of vectors able to express cloned genes efficiently in E. faecalis has hindered biotechnological studies on the bacterium's regulatory and pathogenicity-related genes. The agmatine deiminase (AGDI) pathway of E. faecalis, involved in the conversion of agmatine into putrescine, is driven by a response inducer gene aguR. This study describes that the exposure to the induction factor (agmatine) results in the transcription of genes under the control of the aguB promoter, including the aguBDAC operon. A novel E. faecalis expression vector, named pAGEnt, combining the aguR inducer gene and the aguB promoter followed by a cloning site and a stop codon was constructed. pAGEnt was designed for the overexpression and purification of a protein fused to a 10-amino-acid His-tag at the C-terminus. The use of GFP as a reporter of gene expression in E. faecalis revealed that under induction with 60 mM agmatine, fluorescence reached 40 arbitrary units compared to 0 in uninduced cells. pAGEnt vector can be used for the overexpression of recombinant proteins under the induction of agmatine in E. faecalis, with a close correlation between agmatine concentration and fluorescence when GFP was used as reporter.
Full Text Available Enterococci are considered mainly responsible for the undesirable accumulation of the biogenic amines tyramine and putrescine in cheeses. The biosynthesis of tyramine and putrescine has been described as a species trait in Enterococcus faecalis. Tyramine is formed by the decarboxylation of the amino acid tyrosine, by the tyrosine decarboxylase (TDC route encoded in the tdc cluster. Putrescine is formed from agmatine by the agmatine deiminase (AGDI pathway encoded in the agdi cluster. These biosynthesis routes have been independently studied, tyrosine and agmatine transcriptionally regulate the tdc and agdi clusters. The objective of the present work is to study the possible co-regulation among TDC and AGDI pathways in E. faecalis. In the presence of agmatine, a positive correlation between putrescine biosynthesis and the tyrosine concentration was found. Transcriptome studies showed that tyrosine induces the transcription of putrescine biosynthesis genes and up-regulates pathways involved in cell growth. The tyrosine modulation over AGDI route was not observed in the mutant Δtdc strain. Fluorescence analyses using gfp as reporter protein revealed PaguB (the promoter of agdi catabolic genes was induced by tyrosine in the wild-type but not in the mutant strain, confirming that tdc cluster was involved in the tyrosine induction of putrescine biosynthesis. This study also suggests that AguR (the transcriptional regulator of agdi was implicated in interaction among the two clusters.
Full Text Available Colonizing commensal bacteria after birth are required for the proper development of the gastrointestinal tract. It is believed that bacterial colonization pattern in neonatal gut affects gut barrier function and immune system maturation. Studies on the development of faecal microbiota in infants showed that the neonatal gut was first colonized with enterococci followed by other microbiota such as Bifidobacterium. Other studies showed that babies who developed allergy were less often colonized with Enterococcus during the first month of life as compared to healthy infants. Many studies have been conducted to elucidate how bifidobacteria or lactobacilli, some of which are considered probiotic, regulate infant gut immunity. However, fewer studies have been focused on enterococi. In our study, we demonstrate that E. faecalis, isolated from healthy newborns, suppress inflammatory responses activated in vivo and in vitro. We found E. faecalis attenuates proinflammatory cytokine secretions, especially IL-8, through JNK and p38 signaling pathways. This finding shed light on how the first colonizer, E.faecalis, regulates inflammatory responses in the host.
Full Text Available The effect of dietary supplementation with Enterococcus faecium strain SF68 on growth performance, faecal consistency and in vivo digestibility in buffalo (Bubalus bubalis calves was evaluated. Forty calves were randomly assigned at 10 d of age to one of four treatments: (A milk replacer with no additive, (B milk replacer supplemented with 0.17 g/l of viable (2 x l09 cfu/g E. faecium bacteria daily for 3 days with an interval of 7 days throughout 11 weeks, (C milk replacer supplemented with E. faecium daily for 4 weeks, (D milk replacer supplemented with E. faecium daily for 11 weeks. A total mixed ration was offered ad libitum from 5th week of the experimental period. Faecal score was significantly better in E. faecium-treated calves than control ones. The use of E. faecium had no effect on average daily gain at any stage, total body weight (BW gain, dry matter intake or total tract digestibility. Therefore, E. faecium supplementation may be able to act favourably on the health of the gastrointestinal tract.
Velraeds, M M; van der Mei, H C; Reid, G; Busscher, H J
In this study, 15 Lactobacillus isolates were found to produce biosurfactants in the mid-exponential and stationary growth phases. The stationary-phase biosurfactants from lactobacillus casei subsp. rhamnosus 36 and ATCC 7469, Lactobacillus fermentum B54, and Lactobacillus acidophilus RC14 were investigated further to determine their capacity to inhibit the initial adhesion of uropathogenic Enterococcus faecalis 1131 to glass in a parallel-plate flow chamber. The initial deposition rate of E. faecalis to glass with an adsorbed biosurfactant layer from L. acidophilus RC14 or L. fermentum B54 was significantly decreased by approximately 70%, while the number of adhering enterococci after 4 h of adhesion was reduced by an average of 77%. The surface activity of the biosurfactants and their activity inhibiting the initial adhesion of E. faecalis 1131 were retained after dialysis (molecular weight cutoff, 6,000 to 8,000) and freeze-drying. Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy revealed that the freeze-dried biosurfactants from L. acidophilus RC14 and L. fermentum B54 were richest in protein, while those from L. casei subsp. rhamnosus 36 and ATCC 7469 had relatively high polysaccharide and phosphate contents.
Nueno-Palop, Carmen; Narbad, Arjan
A total of seventy lactic acid bacteria (LAB) were isolated from the faeces of healthy humans and their identities were confirmed by sequencing of their 16S rDNA genes. Of these only 5 isolates were found to resist bile salts and indicated survival in the simulated in vitro digestion assay which reproduces the stomach and intestinal digestion indicating their tolerance to gastric enzymes and the low pH conditions. Species that showed the best resistance to these conditions were: Lactobacillus casei, Lactobacillus sp., uncultured bifidobacteria, Enterococcus faecalis and Streptococcus anginosus. These strains were investigated further to study their capacity to adhere to human intestinal Caco-2 cells. E. faecalis was the most adherent strain. Examination of the virulence determinants for this strain indicated that it was positive for efaAfs, gelE, agg, cpd, cob, ccf and cad, a profile that is similar to that of many E. faecalis isolates from food sources. The cytolysin biosynthetic genes cylA, cylB and cylM that are more associated with the clinical isolates of E. faecium were not detected in this strain. The antibiotic susceptibility tests indicated that the strain was sensitive to vancomycin, tetracycline, rifampicin and erythromycin but resistant only to kanamycin and chloramphenicol. These data suggest that the strain E. faecalis CP58 may be tested further for beneficial properties and developed as a new probiotic. Copyright © 2010 Elsevier B.V. All rights reserved.
Vimont, Allison; Fernandez, Benoît; Hammami, Riadh; Ababsa, Ahlem; Daba, Hocine; Fliss, Ismaïl
Bacterial isolates from raw camel milk were screened for antibacterial activity using the agar diffusion assay. Ten isolates selected for their inhibition of Gram-positive bacteria were identified by 16S sequencing as Enterococcus faecium or durans . An isolate named E. faecium LCW 44 exhibited the broadest antibacterial spectrum with an inhibitory activity against several Gram-positive strains belonging to the genera Clostridium , Listeria , Staphylococcus , and Lactobacillus. E. faecium LCW 44 was shown to produce N-formylated enterocins L50A and L50B, as revealed by mass spectrometry and PCR analyses. This isolate did not harbor any of the virulence factors tested and was shown to be sensitive to all tested antibiotics. It showed high resistance to gastric and intestinal conditions (78 ± 4% survival). Its adhesion index was evaluated at 176 ± 86 and 24 ± 86 on Caco-2 cells and HT-29 cells, respectively, and it significantly reduced adhesion of Listeria monocytogenes by 65 and 49%, respectively. In Macfarlane broth (simulating the nutrient content of the colon), counts of L. monocytogenes were reduced by 2 log 10 cycles after 24 h in co-culture with E. faecium LCW 44, compared to the increase of 4 log 10 cycles when cultured alone. Comparison with a bacteriocin-non-producing mutant of E. faecium LCW 44 strongly suggests that inhibition of L. monocytogenes was due to bacteriocin production. Altogether, E. faecium LCW 44 thus has potential for use as a probiotic for humans and veterinary medicine.
Kishen, Anil; Sum, Chee-Peng; Mathew, Shibi; Lim, Chwee-Teck
Enterococcus faecalis is frequently associated with post-treatment endodontic infections. Because adherence of bacteria to a substrate is the earliest stage in biofilm formation, eliciting the factors that links adherence of this bacterium to dentin would help in understanding its association with treatment-failed root canals. This investigation aimed to study the effects of endodontic irrigants on the adherence of E. faecalis to dentin. The bacteria adherence assay was conducted by using fluorescence microscopy, and the adhesion force was measured by using atomic force microscopy. There were significant increases in adherence and adhesion force after irrigation of dentin with ethylenediaminetetraacetic acid (EDTA), whereas sodium hypochlorite (NaOCl) reduced it. With the use of chlorhexidine (CHX), the force of adhesion increased, but the adherence assay showed a reduction in the number of adhering bacteria. The irrigation regimen of EDTA, NaOCl, and CHX resulted in the least number of adhering E. faecalis cells. This study highlighted that chemicals that alter the physicochemical properties of dentin will influence the nature of adherence, adhesion force, and subsequent biofilm formation of E. faecalis to dentin.
Di Lodovico, Silvia; Cataldi, Valentina; Di Campli, Emanuela; Ancarani, Elisabetta; Cellini, Luigina; Di Giulio, Mara
Nowadays, the bacterial contamination in the hospital environment is of particular concern because the hospital-acquired infections (HAIs), also known as nosocomial infections, are responsible for significant morbidity and mortality. This work evaluated the capability of Enterococcus hirae to form biofilm on different surfaces and the action of two biocides on the produced biofilms. The biofilm formation of E. hirae ATCC 10541 was studied on polystyrene and stainless steel surfaces through the biomass quantification and the cell viability at 20 and 37 °C. The effect of LH IDROXI FAST and LH ENZYCLEAN SPRAY biocides on biomasses was expressed as percentage of biofilm reduction. E. hirae at 20 and 37 °C produced more biofilm on the stainless steel in respect to the polystyrene surface. The amount of viable cells was greater at 20 °C than with 37 °C on the two analyzed surfaces. Biocides revealed a good anti-biofilm activity with the most effect for LH ENZYCLEAN SPRAY on polystyrene and stainless steel at 37 °C with a maximum biofilm reduction of 85.72 and 86.37%, respectively. E. hirae is a moderate biofilm producer depending on surface material and temperature, and the analyzed biocides express a remarkable antibiofilm action. The capability of E. hirae to form biofilm can be associated with its increasing incidence in hospital-acquired infections, and the adoption of suitable disinfectants is strongly recommended.
Higor Fábio Carvalho Bezerra
Full Text Available The objective of this research was to evaluate the effect of whey fermented by Enterococus faecium in consortium with Veilonella parvula in vitro on ruminal microorganisms in different substrates, with or without monensin. The first experiment was carried out in a completely randomized design, in a 6 × 3 factorial arrangement (six substrates × three whey levels with two replicates. In experiment two, a 2 × 3 × 4 factorial arrangement (with and without monensin, three foods and four levels of fermented whey was used, in a randomized design with four replicates, totaling 24 treatments. There was no interaction among the wheys and the substrates in the variable for pectin, starch, and carboxymethyl cellulose. There was a greater growth of amylolytic and pectinolytic microorganisms and a lower growth of proteolytic and cellulolytic microorganisms. A significant effect of optical density was found in the media without substrate and that containing trypticase and glucose due to the addition of fermented whey. There was interaction for the pH at 24 hours among whey, food and monensin. For ammonia at 24 hours there was effect for food, whey and monensin, and interaction among factors. For microbial protein at 24 hours, there was effect for food, whey, monensin and no interaction among sources of variation. The use of whey fermented by bacteria Enterococcus faeciumand Veilonella parvula improves microbial protein synthesis by ruminal bacteria in media containing different energy sources. The combination of fermented whey and monensin shows variable results in relation to microbial growth.
Wulkersdorfer, Beatrix; Jaros, David; Eberl, Sabine; Poschner, Stefan; Jäger, Walter; Cosentini, Enrico; Zeitlinger, Markus; Schwameis, Richard
It has been known from previous studies that body fluids, such as cerebrospinal fluid, lung surfactant, and urine, have a strong impact on the bacterial killing of many anti-infective agents. However, the influence of human bile on the antimicrobial activity of antibiotics is widely unknown. Human bile was obtained and pooled from 11 patients undergoing cholecystectomy. After sterilization of the bile fluid by gamma irradiation, its effect on bacterial killing was investigated for linezolid (LZD) and tigecycline (TGC) against Enterococcus faecalis ATCC 29212. Further, ciprofloxacin (CIP), meropenem (MEM), and TGC were tested against Escherichia coli ATCC 25922. Time-kill curves were performed in pooled human bile and Mueller-Hinton broth (MHB) over 24 h. Bacterial counts (in CFU per milliliter after 24 h) of bile growth controls were approximately equal to MHB growth controls for E. coli and approximately 2-fold greater for E. faecalis , indicating a promotion of bacterial growth by bile for the latter strain. Bile reduced the antimicrobial activity of CIP, MEM, and TGC against E. coli as well as the activity of LZD and TGC against E. faecalis This effect was strongest for TGC against the two strains. Degradation of TGC in bile was identified as the most likely explanation. These findings may have important implications for the treatment of bacterial infections of the gallbladder and biliary tract and should be explored in more detail. Copyright © 2017 American Society for Microbiology.
Yu, Wen; Hallinen, Kelsey; Wood, Kevin
Enterococcus faecalis are commonly associated with hospital acquired infections, because they readily form biofilms on instruments and medical devices. Biofilms are inherently more resistant to killing by antibiotics compared to planktonic bacteria, in part because of their heterogeneous spatial structure. Surprisingly, however, subminimal inhibitory concentrations (sub-MICs) of some antibiotics can actually promote biofilm formation. Unfortunately, much is still unknown about how low drug doses affect the composition and spatial structure of the biofilm. In this work, we investigate the effects of sub-MICs of ampicillin on the formation of E. faecalis biofilms. First, we quantified biofilm mass using crystal violet staining in polystyrene microtiter plates. We found that total biofilm mass is increased over a narrow range of ampicillin concentrations before ultimately declining at higher concentrations. Second, we show that sub-MICs of ampicillin can increase mass of E. faecalis biofilms while simultaneously increasing extracellular DNA/RNA and changing total number of viable cells under confocal microscopy. Further, we use RNA-seq to identify genes differentially expressed under sub-MICs of ampicillin. Finally, we show a mathematical model to explain this phenomenon. This work was funded by The Hartwell Foundation Individual Biomedical Research Award and NSF CAREER 1553208 to KBW.
Mahmoudpour, Ali; Rahimi, Saeed; Sina, Mahmood; Soroush, Mohammad H; Shahi, Shahriar; Shahisa, Shahriar; Asl-Aminabadi, Naser
This study was designed to survey the incidence of Enterococcus faecalis infection in symptomatic and asymptomatic root canals of necrotic teeth using PCR and to isolate the bacterium for further screening. Sixty patients categorized according to their clinical symptoms were used for sampling by insertion of paper points into the root canals and absorbing all the fluids present within them. The samples were incubated in 1.0 ml 2xYT (containing 16 g bacto tryptone, 10 g yeast extract and 5.0 g NaCl per liter) for 24 h at 37 degrees C without aeration prior to multiplex PCR analysis. To assist the isolation of E. faecalis, sub-samples were further grown in the same medium supplemented with 6.5% NaCl and back-inoculated into bile esculin. Using multiple cultivation-dependent and PCR analyses, 6 cases (10%) of E. faecalis were identified. Four isolates were obtained from asymptomatic cases of chronic apical periodontitis, and the other two were associated with phoenix abscess and acute apical abscess, respectively. No E. faecalis infection was found in 5 patients with acute apical periodontitis or in 9 with chronic suppurative periodontitis. Our results indicate that there is no significant difference in the incidence of E. faecalis between symptomatic and asymptomatic necrotic dental root canals (P > 0.05).
Full Text Available Enterococcus faecalis is a Gram-positive lactic acid intestinal opportunistic bacterium with virulence potential. For a better understanding of the adapation of this bacterium to the host conditions, we performed a transcriptome analysis of bacteria isolated from an infection site (mouse peritonitis by RNA-sequencing. We identified a total of 211 genes with significantly higher transcript levels and 157 repressed genes. Our in vivo gene expression database reflects well the infection process since genes encoding important virulence factors like cytolysin, gelatinase or aggregation substance as well as stress response proteins, are significantly induced. Genes encoding metabolic activities are the second most abundant in vivo induced genes demonstrating that the bacteria are metabolically active and adapt to the special nutrient conditions of the host. α- and β- glucosides seem to be important substrates for E. faecalis inside the host. Compared to laboratory conditions, the flux through the upper part of glycolysis seems to be reduced and more carbon may enter the pentose phosphate pathway. This may reflect the need of the bacteria under infection conditions to produce more reducing power for biosynthesis. Another important substrate is certainly glycerol since both pathways of glycerol catabolism are strongly induced. Strongly in vivo induced genes should be important for the infection process. This assumption has been verified in a virulence test using well characterized mutants affected in glycerol metabolism. This showed indeed that mutants unable to metabolize this sugar alcohol are affected in organ colonisation in a mouse model.
Alshareef, A.; Laird, K.; Cross, R. B. M.
Silver nanoparticles (AgNPs) have been shown to exhibit strong antibacterial activity against both Gram-positive bacteria and Gram-negative bacteria including antibiotic resistant strains. This study aims to compare the bactericidal effect of different shaped AgNPs (spherical and truncated octahedral) against Escherichia coli and Enterococcus faecium. The antimicrobial activity of a range of concentrations (50, 100, 1000 μg/ml) was determined over 24 h using both optical density and viable counts. Truncated octahedral AgNPs (AgNOct) were found to be more active when compared with spherical AgNPs (AgNS). The difference in shape resulted in differences in efficacy which may be due to the higher surface area of AgNOct compared to AgNS, and differences in active facets and surface energies, with AgNPs having a bacteriostatic effect and AgNOct being bactericidal after 4 h. The results suggest that AgNPs can be used as effective growth inhibitors in different microorganisms, rendering them applicable to various medical devices and antimicrobial control systems.
Bucci, Vanni; Caballero, Silvia; Djukovic, Ana; Toussaint, Nora C.; Equinda, Michele; Lipuma, Lauren; Ling, Lilan; Gobourne, Asia; No, Daniel; Taur, Ying; Jenq, Robert R.; van den Brink, Marcel R. M.; Xavier, Joao B.
Bacteria causing infections in hospitalized patients are increasingly antibiotic resistant. Classical infection control practices are only partially effective at preventing spread of antibiotic-resistant bacteria within hospitals. Because the density of intestinal colonization by the highly antibiotic-resistant bacterium vancomycin-resistant Enterococcus (VRE) can exceed 109 organisms per gram of feces, even optimally implemented hygiene protocols often fail. Decreasing the density of intestinal colonization, therefore, represents an important approach to limit VRE transmission. We demonstrate that reintroduction of a diverse intestinal microbiota to densely VRE-colonized mice eliminates VRE from the intestinal tract. While oxygen-tolerant members of the microbiota are ineffective at eliminating VRE, administration of obligate anaerobic commensal bacteria to mice results in a billionfold reduction in the density of intestinal VRE colonization. 16S rRNA gene sequence analysis of intestinal bacterial populations isolated from mice that cleared VRE following microbiota reconstitution revealed that recolonization with a microbiota that contains Barnesiella correlates with VRE elimination. Characterization of the fecal microbiota of patients undergoing allogeneic hematopoietic stem cell transplantation demonstrated that intestinal colonization with Barnesiella confers resistance to intestinal domination and bloodstream infection with VRE. Our studies indicate that obligate anaerobic bacteria belonging to the Barnesiella genus enable clearance of intestinal VRE colonization and may provide novel approaches to prevent the spread of highly antibiotic-resistant bacteria. PMID:23319552
Full Text Available Bacterial isolates from raw camel milk were screened for antibacterial activity using the agar diffusion assay. Ten isolates selected for their inhibition of Gram-positive bacteria were identified by 16S sequencing as Enterococcus faecium or durans. An isolate named E. faecium LCW 44 exhibited the broadest antibacterial spectrum with an inhibitory activity against several Gram-positive strains belonging to the genera Clostridium, Listeria, Staphylococcus, and Lactobacillus. E. faecium LCW 44 was shown to produce N-formylated enterocins L50A and L50B, as revealed by mass spectrometry and PCR analyses. This isolate did not harbor any of the virulence factors tested and was shown to be sensitive to all tested antibiotics. It showed high resistance to gastric and intestinal conditions (78 ± 4% survival. Its adhesion index was evaluated at 176 ± 86 and 24 ± 86 on Caco-2 cells and HT-29 cells, respectively, and it significantly reduced adhesion of Listeria monocytogenes by 65 and 49%, respectively. In Macfarlane broth (simulating the nutrient content of the colon, counts of L. monocytogenes were reduced by 2 log10 cycles after 24 h in co-culture with E. faecium LCW 44, compared to the increase of 4 log10 cycles when cultured alone. Comparison with a bacteriocin-non-producing mutant of E. faecium LCW 44 strongly suggests that inhibition of L. monocytogenes was due to bacteriocin production. Altogether, E. faecium LCW 44 thus has potential for use as a probiotic for humans and veterinary medicine.
Nami, Yousef; Abdullah, Norhafizah; Haghshenas, Babak; Radiah, Dayang; Rosli, Rozita; Khosroushahi, Ahmad Yari
Forty-five lactic acid bacteria (LAB) were isolated from the vaginal specimens of healthy fertile women, and the identities of the bacteria were confirmed by sequencing of their 16S rDNA genes. Among these bacteria, only four isolates were able to resist and survive in low pH, bile salts and simulated in vitro digestion conditions. Lactococcus lactis 2HL, Enterococcus durans 6HL, Lactobacillus acidophilus 36YL and Lactobacillus plantarum 5BL showed the best resistance to these conditions. These strains were evaluated further to assess their ability to adhere to human intestinal Caco-2 cells. Lactococcus lactis 2HL and E. durans 6HL were the most adherent strains. In vitro tests under neutralized pH proved the antimicrobial activity of both strains. Results revealed that the growth of Escherichia coli O26, Staphylococcus aureus and Shigella flexneri was suppressed by both LAB strains. The antibiotic susceptibility tests showed that these strains were sensitive to all nine antibiotics: vancomycin, tetracycline, ampicillin, penicillin, gentamicin, erythromycin, clindamycin, sulfamethoxazole and chloramphenicol. These data suggest that E. durans 6HL and Lactococcus lactis 2HL could be examined further for their useful properties and could be developed as new probiotics. © 2014 The Authors.
Morgan, Daniel J; Murthy, Rekha; Munoz-Price, L Silvia; Barnden, Marsha; Camins, Bernard C; Johnston, B Lynn; Rubin, Zachary; Sullivan, Kaede V; Shane, Andi L; Dellinger, E Patchen; Rupp, Mark E; Bearman, Gonzalo
Whether contact precautions (CP) are required to control the endemic transmission of methicillin-resistant Staphylococcus aureus (MRSA) or vancomycin-resistant Enterococcus (VRE) in acute care hospitals is controversial in light of improvements in hand hygiene, MRSA decolonization, environmental cleaning and disinfection, fomite elimination, and chlorhexidine bathing. To provide a framework for decision making around use of CP for endemic MRSA and VRE based on a summary of evidence related to use of CP, including impact on patients and patient care processes, and current practices in use of CP for MRSA and VRE in US hospitals. A literature review, a survey of Society for Healthcare Epidemiology of America Research Network members on use of CP, and a detailed examination of the experience of a convenience sample of hospitals not using CP for MRSA or VRE. Hospital epidemiologists and infection prevention experts. No high quality data support or reject use of CP for endemic MRSA or VRE. Our survey found more than 90% of responding hospitals currently use CP for MRSA and VRE, but approximately 60% are interested in using CP in a different manner. More than 30 US hospitals do not use CP for control of endemic MRSA or VRE. Higher quality research on the benefits and harms of CP in the control of endemic MRSA and VRE is needed. Until more definitive data are available, the use of CP for endemic MRSA or VRE in acute care hospitals should be guided by local needs and resources.
Ana María Sánchez-Díaz
Full Text Available Enterococcus faecium and Streptococcus gallolyticus subsp. gallolyticus (S. gallolyticus were classically clustered into the Lancefield Group D streptococci and despite their taxonomic reclassification still share a similar genetic content and environment. Both species are considered as opportunistic pathogens. E. faecium is often associated with nosocomial bacteraemia, and S. gallolyticus is sporadically found in endocarditis of colorectal cancer patients. In both cases, the source of infection is commonly endogenous with a translocation process that launches through the intestinal barrier. To get new insights into the pathological processes preceding infection development of both organisms, we used an in vitro model with Caco-2 cells to study and compare the adhesion, invasion and translocation inherent abilities of 6 E. faecium and 4 S. gallolyticus well-characterized isolates. Additionally, biofilm formation on polystyrene, collagen I and IV was also explored. Overall results showed that E. faecium translocated more efficiently than S. gallolyticus, inducing a destabilization of the intestinal monolayer. Isolates Efm106, Efm121 and Efm113 (p < .001 compared to Ef222 exhibited the higher translocation ability and were able to adhere 2-3 times higher than S. gallolyticus isolates. Both species preferred the collagen IV coated surfaces to form biofilm but the S. gallolyticus structures were more compact (p = .01. These results may support a relationship between biofilm formation and vegetation establishment in S. gallolyticus endocarditis, whereas the high translocation ability of E. faecium high-risk clones might partially explain the increasing number of bacteraemia.
Khine Zar Wynn Myint; Khin Mya Lwin; Myo Myint
The microbial gibberellic acid (GA3) production of Pseudomonas spp., was studied and qualitatively indentified by UV spectrophotometer. 20 strains of Pseudomonas spp., were isolated and screened the gibberellic acid productivily in King's B medium. Among them, only four strains can produce microbial gibberellic acid. The Rf values and colour appearance under UV were the same as authentic gibberellic acid. Moreover, the gibberellic acid producer strains were identified as Pseudomonas spp., by cultural, biochemical and drug sensitivity pattern.
Full Text Available Valeriana spp. is a flowering plant that is well known for its essential oils, iridoid compounds such as monoterpenes and sesquiterpenes, flavonoids, alkaloids, amino acids, and lignanoids. Valeriana spp. exhibits a wide range of biological activities such as lowering blood pressure and heart rate, antimyocardial ischemia reperfusion injury, antiarrhythmia, and regulation of blood lipid levels. This review focuses on the chemical constituents and cardiovascular activities of Valeriana spp.
Doni, Febri; Anizan, I.; Che Radziah C. M., Z.; Yusoff, Wan Mohtar Wan
Trichoderma spp. are considered as successful plant growth promoting fungi and have positive role in habitat engineering. In this study, the potential for Trichoderma spp. to regulate transpiration process in rice plant was assessed experimentally under greenhouse condition using a completely randomized design. The study revealed that Trichoderma spp. have potential to enhance growth of rice plant through transpirational processes. The results of the study add to the advancement of the understanding as to the role of Trichoderma spp. in improving rice physiological process.
... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3375 Mycoplasma... fluorescent dye (immunofluorescent reagents) used to identify Mycoplasma spp. directly from clinical specimens...
Slovensky plynarensky priemysel (SPP) officially indicated that it intended to drop its project of using geothermal energy in the Kosicka kotlina. This spring it published an advert that it was looking for a company that wished to acquire a majority stake in the company, Geoterm Kosice. The company was established to commercially develop this geothermal source. But it seems SPP does not want to drop the project completely. It has kept some important cards, such as control over the land where the boreholes are located Any company that wants to use geothermal energy needs a ruling issued by the Ministry of Environment defining the exploration area. Geothermal sources were found in the villages of Durkov, Svinica, Bidovce and Olsovany. Not so long ago the area was assigned to Geoterm but from May 9 the area can be explored by Slovgeoterm. Both companies have the same majority shareholder - SPP. It controls 96% of Geoterm shares and 50% of Slovgeoterm. So far it has only officially announced its intention to sell the Geoterm shares. But as far as the use of the geothermal resource is concerned since May Slovgeoterm has played a key role.The company focuses on the utilization of geothermal energy. In addition to the project in the Kosice region, it has also participated in a project to heat more than a thousand flats using geothermal water in Galanta and a project to heat greenhouses in Podhajske. There are also other geothermal projects running in Presov and Michalovce. Icelandic company, Enex, with the same specialisation controls 28% of the company and a further 20% is owned by the investment group, NEFCO based in Helsinki. Two percent of the company is owned by its general director and the general proxy of Geoterm, Otto Halas. And so without the agreement of this company no-one can start any activities related to the utilization of geothermal energy. (authors)
The genus of Phytophthora is a destructive plants pathogen. However the existence of these species in plant tissue was hardly to detect because the fungus may also be present as resistant propagules in soil or spread through waterways. This study aimed to test the FTA card as a direct bait to obtain the DNA of Phytophthora spp and also to investigate the suitability of FTA card as a sampling method. This research used several Phytophthora baits including FTA card followed by DNA extraction a...
Shi, Xiang; Wang, Shufeng; Sun, Haijing; Chen, Yitai; Wang, Dongxue; Pan, Hongwei; Zou, Yazhu; Liu, Jianfeng; Zheng, Linyu; Zhao, Xiulian; Jiang, Zeping
A pot experiment was conducted to evaluate the feasibility of using tree seedlings for the phytoremediation of lead/zinc (Pb/Zn) mine tailings. Seedlings of three Quercus spp. (Q. shumardii, Q. phellos, and Q. virginiana) and rooted cuttings of two Salix spp. (S. matsudana and S. integra) were transplanted into pots containing 50 and 100 % Pb/Zn mine tailings to evaluate their tolerance of heavy metals. The five species showed different tolerance levels to the Pb/Zn tailings treatments. Q. virginiana was highly tolerant to heavy metals and grew normally in the Pb/Zn tailings. The root systems showed marked differences between the Quercus spp. and Salix spp., indicating that different mechanisms operated to confer tolerance of heavy metals. The maximum efficiency of photosystem II photochemistry value of the five species showed no differences among the treatments, except for Q. shumardii. All species showed low metal translocation factors (TFs). However, S. integra had significantly higher TF values for Zn (1.42-2.18) and cadmium (1.03-1.45) than did the other species. In this respect, Q. virginiana showed the highest tolerance and a low TF, implying that it is a candidate for phytostabilization of mine tailings in southern China. S. integra may be useful for phytoextraction of tailings in temperate regions.
Center, Barbara J.; Giblin-Davis, Robin M.; Herre, E. Allen; Chung-Schickler, Genevieve C.
Syconia (enclosed infructescences) infested with host-specific species of Schistonchus (Aphelenchoididae) were collected from six species of Ficus (Moraceae) native to Florida or Panama. They were sectioned and histologically examined to assess the effects of parasitism. Parasitism by Schistonchus spp. was associated with hypertrophied cells, tissue necrosis, and the presence of an exudate in all species. Occasional hypertrophy of the outer epidermal cells occurred on seed florets, wasp florets, and on the endothecial cells of male florets in F. aurea (subgenus Urostigma) from Florida. Aberrations of the inner mesocarp occurred under the hypertrophied cells on seed florets. In F. laevigata (subgenus Urostigma) from Florida, Schistonchus sp. infested immature male florets and was associated with hypertrophy of endothecial cells, epidermal cells of the anther filaments, and anthers. Schistonchus sp. also caused aberrations of the anther filament, anthers, and pollen. Ficus poponoei (subgenus Urostigma) and F. glabrata (subgenus Pharmacosycea), both from Panama, had hypertrophied outer epidermal cells on seed florets. Ficus poponoei also had Schistonchus sp. within the pedicel of an aborted floret, with hypertrophy of the cortical parenchyma. Ficus trigonata (subgenus Urostigma) from Panama had hypertrophy of the outer epidermis of seed florets. When the outer epidermis on these florets was missing, the inner mesocarp was hypertrophied. Ficus maxima (subgenus Pharmacosycea) from Panama had hypertrophy on the outer epidermis of seed and aborted florets. Schistonchus spp. were not found in wasp larvae or pupae in any of the Ficus spp. examined. Hypertrophy was never observed in the absence of Schistonchus spp. PMID:19270912
Full Text Available Blackish or orange liquid-like spots were found on (n=100 fruits of chillies (Capsicum sold in five local markets in Kota Kinabalu, Sabah, Malaysia. Colletotrichum gloeosporioides and C. capsici were identified as the causal agents of an anthracnose disease. This is the first report of Colletotrichum spp. as the causal agent of anthracnose infected chillies in Sabah.
Marcielly F. Turatto
Full Text Available ABSTRACT Plant Growth Promoting Rhizobacteria (PGPR have different mechanisms of action in the development of plants, such as growth promotion, production of phytohormones and antibiotic substances and changes in root exudates. These help to control plant diseases. In order to evaluate the potential of microorganisms in the control of Meloidogyne javanica and Ditylenchus spp., five rhizobacteria isolated from rhizosphere of garlic cultivated in the Curitibanos (SC region were tested. Hatching chambers were set on Petri dishes, in which were added 10 mL of bacterial suspension and 1 mL of M. javanica eggs suspension, at the rate of 4500, on the filter paper of each chamber. The same procedure was performed with 300 juvenile Ditylenchus spp. The experimental design was completely randomized, with four replications. The evaluations were performed every 72 h for nine days. The antagonized population of nematodes was determined in Peters counting chamber, determining the percentage hatching (for M. javanica and motility (for Ditylenchus spp. Isolates CBSAL02 and CBSAL05 significantly reduced the hatching of M. javanica eggs (74% and 54.77%, respectively and the motility of Ditylenchus spp. (55.19% and 53.53%, respectively in vitro. Isolates were identified as belonging to the genera Pseudomonas (CBSAL05 and Bacillus (CBSAL02.
Furustrand Tafin, Ulrika; Meis, Jacques F; Trampuz, Andrej
We evaluated isothermal microcalorimetry for real-time susceptibility testing of non-Aspergillus molds. MIC and minimal effective concentration (MEC) values of Mucorales (n = 4), Fusarium spp. (n = 4), and Scedosporium spp. (n = 4) were determined by microbroth dilution according to the Clinical Laboratory Standard Institute M38-A2 guidelines. Heat production of molds was measured at 37 °C in Sabouraud dextrose broth inoculated with 2.5 × 10(4) spores/mL in the presence of amphotericin B, voriconazole, posaconazole, caspofungin, and anidulafungin. As determined by microcalorimetry, amphotericin B was the most active agent against Mucorales (MHIC 0.06-0.125 μg/mL) and Fusarium spp. (MHIC 1-4 μg/mL), whereas voriconazole was the most active agent against Scedosporium spp. (MHIC 0.25 to 8 μg/mL). The percentage of agreement (within one 2-fold dilution) between the MHIC and MIC (or MEC) was 67%, 92%, 75%, and 83% for amphotericin B, voriconazole, posaconazole, and caspofungin, respectively. Microcalorimetry provides additional information on timing of antifungal activity, enabling further investigation of drug-mold and drug-drug interaction, and optimization of antifungal treatment. Copyright © 2012 Elsevier Inc. All rights reserved.
Margalit Levi, Maayan; Nachum-Biala, Yaarit; King, Roni; Baneth, Gad
Babesia spp. and Hepatozoon spp. are apicomplexan parasites that infect a variety of animals, including canids. Their life-cycle includes an invertebrate hematophagous vector as a definitive host and vertebrates as intermediate hosts. The aims of this study were to investigate the prevalence and risk factors for Babesia spp. and Hepatozoon spp. infections in wild golden jackals (Canis aureus) and red foxes (Vulpes vulpes) in Israel and to compare spleen with blood sample polymerase chain reaction (PCR) for the detection of infection. Blood and spleen samples from 109 golden jackals and 21 red foxes were tested by PCR for the detection of Babesia spp. and Hepatozoon spp. using primers for the 18S ribosomal (r) RNA gene. Hepatozoon canis was detected in 50/109 (46%) of the jackals and 9/21 (43%) of the foxes. "Babesia vulpes" (the Babesia microti-like piroplasm) was detected in 4/21 (19%) of the foxes and in none of the jackals. A previously unknown genotype termed Babesia sp. MML related to Babesia lengau (96-97% identity) was detected in 1/109 (1%) of the jackals and 4/21 (19%) of the foxes. Further characterization of this genotype carried out by PCR of the rRNA internal transcribed spacer 2 (ITS2) indicated that it had only 87% identity with the B. lengau ITS2. Sex (male or female), age (juvenile or adult) and geographic zone (North, Central or South Israel) were not found to be significant risk factors for these protozoan infections. The prevalence of "B. vulpes" and Babesia sp. MML infections was significantly higher in foxes compared to jackals (χ 2 = 15.65, df = 1, P < 0.005), while there was no statistically significant difference in the rate of H. canis infection between these two canid species. A fair agreement beyond chance between identification in the blood and spleen of H. canis was found in 21 animals from which both blood and spleen samples were available (k = 0.33). This study describes a high prevalence of H. canis infection in
..., tonsillitis, and bronchitis due to Streptococcus spp., Staphylococcus spp., Escherichia coli, Proteus mirabilis, and Pasteurella spp., urinary tract infections (cystitis) due to Streptococcus spp., Staphylococcus spp., E., coli, P. mirabilis, and Enterococcus spp.; gastrointestinal infections due to...
Stefanie Bressan Waller
Full Text Available ABSTRACT Cases of sporotrichosis in humans and animals without satisfactory clinical response have increased, a warning sign of strains resistant to conventional antifungal agents. The urgent search for alternative therapies was an incentive for research on medicinal plants with anti-Sporothrix spp. properties. A bibliographic survey was performed based on scientific papers about in vitro and in vivo antifungal activity of essential oils and extracts of plants in differents solvents against the fungal of the Sporothrix schenckii complex. The study methodology consisted of a literature review in Google Scholar, Science Direct, Pubmed, Bireme and Springer link with papers from 1986 to 2015. We found 141 species of plants that were investigated, of which 100 species were concentrated in 39 botanical families that had confirmed anti-Sporothrix activity. Combretaceae, Asteraceae and Lamiaceae represented the botanical families with the greatest number of plants species with antifungal potential, using different methodologies. However, there are few studies with medicinal plants in experimental infection in animals that prove their activity in the treatment of sporotrichosis. It reinforces the need for further research related to standardization of in vitro methodologies and in vivo studies related to safety and to toxicity potential of these plants with anti-Sporothrix spp. activity.
Hirt, Helmut; Greenwood-Quaintance, Kerryl E; Karau, Melissa J; Till, Lisa M; Kashyap, Purna C; Patel, Robin; Dunny, Gary M
Cell-cell communication mediated by peptide pheromones (cCF10 [CF]) is essential for high-frequency plasmid transfer in vitro in Enterococcus faecalis To examine the role of pheromone signaling in vivo , we established either a CF-producing (CF+) recipient or a recipient producing a biologically inactive variant of CF (CF- recipient) in a germfree mouse model 3 days before donor inoculation and determined transfer frequencies of the pheromone-inducible plasmid pCF10. Plasmid transfer was detected in the upper and middle sections of the intestinal tract 5 h after donor inoculation and was highly efficient in the absence of antibiotic selection. The transconjugant/donor ratio reached a maximum level approaching 1 on day 4 in the upper intestinal tract. Plasmid transfer was significantly lower with the CF- recipient. While rescue of the CF- mating defect by coculture with CF+ recipients is easily accomplished in vitro , no extracellular complementation occurred in vivo This suggests that most pheromone signaling in the gut occurs between recipient and donor cells in very close proximity. Plasmid-bearing cells (donors plus transconjugants) steadily increased in the population from 0.1% after donor inoculation to about 10% at the conclusion of the experiments. This suggests a selective advantage of pCF10 carriage distinct from antibiotic resistance or bacteriocin production. Our results demonstrate that pheromone signaling is required for efficient pCF10 transfer in vivo In the absence of CF+ recipients, a low level of transfer to CF- recipients occurred in the gut. This may result from low-level host-mediated induction of the donors in the gastrointestinal (GI) tract, similar to that previously observed in serum. IMPORTANCE Horizontal gene transfer is a major factor in the biology of Enterococcus faecalis , an important nosocomial pathogen. Previous studies showing efficient conjugative plasmid transfer in the gastrointestinal (GI) tracts of experimental animals did
Full Text Available Background: Periapical lesions, are characterized by an immune response to the invading bacteria consequences periapical bone destruction. In root canal treatment failure was found Enterococcus faecalis (E. faecalis as most species. iNOS found an important role in protection against infection, plays vital roles in fighting pathogens and contributing to disease pathology. Purpose: This study was to observed the role of iNOS in teeth periapical lessions immunopathogenesis caused by E. faecalis. Methods: The randomized post-only control group design used in this study, This study used 24 Wistar rats, were divided into three groups (each group consisted of 8 rats, as negative controls group is a normal teeth, in the positive controls group was made by drilling the upper right first molar to penetrate the dental pulp and was induced with 10µl BHI-b then filled with Glass Ionomer Cement (GIC and the treatment group, after drilling the teeth, then inoculated with E. faecalis ATCC 29212 106 CFU into 10µl BHI-b then filled with GIC to prevent contamination. It takes 21 days to get periapical lesions and rat were sacrificed, and then the expression of iNOS was measured. Results: Statistical analysis using ANOVA found a significant differenced between control and treatment groups (p<0.05. Conclusion: This study concluded that iNOS role in teeth periapical lesions immunopathogenesis caused by E. faecalis.Latar belakang: Lesi periapikal merupakan hasil suatu respon imun untuk melawan invasi bakteri yang mengakibatkan destruksi tulang periapikal. Pada perawatan saluran akar yang mengalami kegagalan ditemukan Enterococcus faecalis sebagai spesies terbanyak. iNOS berperan penting untuk proteksi terhadap bakteri, mempunyai peran yang vital untuk melawan patogen dan berkonstribusi secara patologik untuk menyebabkan suatu penyakit. Tujuan: Penelitian ini bertujuan untuk mengobservasi peran iNOS secara imunohistokimia pada lesi periapikal tikus Wistar. Metode
Baron, Aleksandr; Lindsey, Kimberly; Sidow, Stephanie J; Dickinson, Douglas; Chuang, Augustine; McPherson, James C
The purpose of this investigation was to determine the effect of a sodium hypochlorite-surfactant combination on the removal of Enterococcus faecalis from infected teeth. Sixty-four extracted human single canal anterior teeth were prepared with rotary instrumentation and sterilized. Teeth were divided into 4 groups, N = 16. Three experimental groups were inoculated with E. faecalis and cultured for 21 days before use: positive control group, no irrigation; NaOCl group, irrigated with 5 mL 6% NaOCl; and NaOCl/BAK group, irrigated with 5 mL 6% NaOCl/0.008% benzalkonium chloride (BAK). The negative control group received medium only and no inoculate. Paper point sampling of the canals was obtained before irrigation (S1) for all 4 groups and for 2 groups after irrigation (S2) to determine remaining colony-forming units. After sampling, all teeth were split in half and evaluated for bacterial viability colony-forming units and penetration of dentinal tubules by using fluorescent vital dye staining and confocal laser scanning microscopy. Comparison of pre-irrigation and post-irrigation paper point samples from the 2 irrigated groups showed a significant reduction in bacterial canal load (P < .001, Kruskal-Wallis), with a significantly lower load in the NaOCl/BAK group than in the NaOCl group (P = .001, Mann-Whitney U test); 68.8% of the NaOCl/BAK samples gave no recoverable counts. In contrast, no significant difference between these groups was found for counts recovered from dentin. Confocal laser scanning microscopy showed no differences in tubule penetration. The addition of BAK to NaOCl significantly reduced the number of remaining bacteria within the canal after irrigation compared with NaOCl alone. Published by Elsevier Inc.
Tatiana Kelly da Silva Fidalgo
Full Text Available The present study evaluated the antimicrobial activity of three root canal irrigants against Enterococcus faecalis, Candida albicans, and Staphylococcus aureus. These microorganisms were incubated in the presence of citric acid (6 and 10%, EDTA (17%, and NaOCl (0.5, 1.0, 2.5, and 5.25%. Agar diffusion tests were performed and redox indicator resazurin was used to evaluate the inhibitory effect of the irrigants on the metabolic activity of these microorganisms. The mean diameters of the inhibition zones for the C. albicans cultures were 11.6 mm (17% EDTA, 5.5 mm (0.5% NaOCl, 12.9 mm (1% NaOCl, 22.1 mm (2.5% NaOCl, and 28.5 mm (5.25% NaOCl. The mean diameters of the inhibition zones for E. faecalis were 2.8 mm (1% NaOCl, 5.4 mm (2.5% NaOCl, and 8.3 mm (5.25% NaOCl. For S. aureus, the mean values were 8.0 mm (17% EDTA, 3.0 mm (1% NaOCl, 8.8 mm (2.5% NaOCl, and 10.0 mm (5.25% NaOCl. Most of the irrigant solutions presented effective antimicrobial activity against C. albicans. A high inhibitory effect on the metabolic activity of E. faecalis was detected when the microorganisms were incubated with 17% EDTA. The same result was reached when S. aureus was incubated in the presence of > 2.5% NaOCl. Altogether, these results indicate that 2.5% and 5.25% NaOCl are microbicides against S. aureus while 0.5% and 1% NaOCl are only microbiostatic against the tested bacteria. The 6% and 10% citric acid as well as 17% EDTA did not affect the viability of any of the assayed microorganisms.
Maekawa, Lilian Eiko; Rossoni, Rodnei Dennis; Barbosa, Júnia Oliveira; Jorge, Antonio Olavo Cardoso; Junqueira, Juliana Campos; Valera, Marcia Carneiro
Dried, fresh and glycolic extracts of Zingiber officinale were obtained to evaluate the action against G. mellonella survival assay against Enterococcus faecalis infection. Eighty larvae were divided into: 1) E. faecalis suspension (control); 2) E. faecalis + fresh extract of Z. officinale (FEO); 3) E. faecalis + dried extract of Z. officinale (DEO); 4) E. faecalis + glycolic extract of Z. officinale (GEO); 5) Phosphate buffered saline (PBS). For control group, a 5 μL inoculum of standardized suspension (107 cells/mL) of E. faecalis (ATCC 29212) was injected into the last left proleg of each larva. For the treatment groups, after E. faecalis inoculation, the extracts were also injected, but into the last right proleg. The larvae were stored at 37 °C and the number of dead larvae was recorded daily for 168 h (7 days) to analyze the survival curve. The larvae were considered dead when they did not show any movement after touching. E. faecalis infection led to the death of 85% of the larvae after 168 h. Notwithstanding, in treatment groups with association of extracts, there was an increase in the survival rates of 50% (GEO), 61% (FEO) and 66% (DEO) of the larvae. In all treatment groups, the larvae exhibited a survival increase with statistically significant difference in relation to control group (p=0.0029). There were no statistically significant differences among treatment groups with different extracts (p=0.3859). It may be concluded that the tested extracts showed antimicrobial activity against E. faecalis infection by increasing the survival of Galleria mellonella larvae.
Full Text Available Enterococcus faecalis is a natural inhabitant of the human gastrointestinal tract. However, as an opportunistic pathogen, it is able to colonize other host niches and cause life-threatening infections. Its adaptation to new environments involves global changes in gene expression. The EF3013 gene (here named mafR of E. faecalis strain V583 encodes a protein (MafR, 482 residues that has sequence similarity to global response regulators of the Mga/AtxA family. The enterococcal OG1RF genome also encodes the MafR protein (gene OG1RF_12293. In this work, we have identified the promoter of the mafR gene using several in vivo approaches. Moreover, we show that MafR influences positively the transcription of many genes on a genome-wide scale. The most significant target genes encode components of PTS-type membrane transporters, components of ABC-type membrane transporters, and proteins involved in the metabolism of carbon sources. Some of these genes were previously reported to be up-regulated during the growth of E. faecalis in blood and/or in human urine. Furthermore, we show that a mafR deletion mutant strain induces a significant lower degree of inflammation in the peritoneal cavity of mice, suggesting that enterococcal cells deficient in MafR are less virulent. Our work indicates that MafR is a global transcriptional regulator. It might facilitate the adaptation of E. faecalis to particular host niches and, therefore, contribute to its potential virulence.
Armelia Sari Widyarman
Full Text Available Background: Enterococcus faecalis (E. faecalis and Porphyromonas gingivalis (P. gingivalis are oral bacteria related to root canal infection and periodontal disease pathogenesis. Strawberries (Fragaria x ananassa fruit are rich in vitamins and minerals, have antibacterial and antioxidant effects. Objective: This study investigated the inhibition effect of strawberry extract on monospecies and multispecies E. faecalis and P. gingivalis bacteria grown as biofilms in vitro. Methods: This study used E. faecalis ATCC 29212 and P. gingivalis ATCC 33277. It analyzed the effect of strawberry extract on bacteria biofilm formation using a biofilm assay on microplate wells. Five concentrations of strawberry extracts were used (100%, 50%, 25%, 12.5%, and 6.25%, and the inhibition effect was observed after a 1h, 3h, 6h, and 24h incubation period. Biofilms without the strawberry extract were used as the negative controls, and crystal violet and safranin (0.5%w/v were used to count the biofilm mass. The biofilms grown on microplates were counted using an ELISA reader at 450 nm after 200 mL of 90% ethanol was added to attract the absorbed stain. The strawberry extract inhibition effectiveness on the biofilm formation of each bacterium tested was analyzed using one-way Anova, where p<0.05 was defined as a significant difference. Result: The strawberry extract inhibited the tested monospecies and multispecies bacteria biofilm formation. The optimal strawberry extract concentration for the inhibition of either monospecies biofilms was 100%. However, the optimal incubation time for the strawberry extract to inhibit the multispecies biofilm formation was 24h, which was the study’s biofilm maturity phase. Conclusions: The 100% strawberry extract concentration inhibited the formation of both the monospecies and multispecies E. faecalis and P. gingivalis biofilms. Future studies are needed to evaluate the potential of strawberry extract as an alternative dental
Adriana Lígia de Castilho
Full Text Available Enterococcus faecalis is an important pathogen associated with endodontic diseases, and its elimination and control are of paramount importance, as it represents one of the major causes of failure in the treatment of endodontic disease. Twenty-five plant extracts obtained from Brazilian forests were found to be effective against planktonic E. faecalis and were subjected to two traditional antibacterial assays, the microdilution broth assay (MDBA and the disk diffusion assay (DDA, using chlorhexidine (CHX as a control. Seven out of 25 extracts showed significant antibacterial activity and were tested in a biofilm assay, and three of these extracts were subjected to chemical fractionation. Residues were tested for their antibacterial activity, and the first chemical findings were described based on thin layer chromatography (TLC. Extracts obtained from Ipomoea alba, Symphonia globulifera and Moronobea coccinea showed significant bactericidal activity in the MDBA. The same I. alba and S. globulifera extracts, as well as the extract obtained from Connarus ruber var. ruber, showed significant activity in the DDA. RH2O obtained from Psidium densicomum and Stryphnodendron pulcherrimum showed better antibacterial activity compared to the respective crude extracts and CHX. TLC analysis showed that phenolic compounds and triterpenes represent the first findings of chemical groups that may occur in all species. The results of the present study include the discovery of six active extracts against planktonic E. faecalis and support further testing via assays involving biofilm formation, as well as the determination of the compounds' chemical profiles, as their activity was significantly better than that observed for CHX.
de Almeida, Josiane; Hoogenkamp, Michel; Felippe, Wilson T; Crielaard, Wim; van der Waal, Suzette V
Disruption of the matrix of endodontic biofilms will aid in their removal from a root canal. Therefore, the aim of this study was to investigate the efficacy of EDTA and a modified salt solution (MSS) to detach bacteria from biofilms. Forty-eight-hour-old Enterococcus faecalis biofilms were grown on glass coverslips and then treated for 1 hour by immersion in 17% EDTA or MSS. Phosphate-buffered saline served as a negative control. Then, residual biofilm cells on the substrate and the detached cells in the supernatant were collected. Viability was verified by the colony-forming unit (CFU) counting method. Propidium monoazide (PMA) treatment in conjunction with quantitative polymerase chain reaction (qPCR) was also performed to detect the presence of E. faecalis 16S ribonucleic RNA genes. Data were analyzed using 1-way analysis of variance and Tukey or Kruskal-Wallis and Dunn tests. The Pearson R test evaluated the correlation between results from CFU and PMA (α = 5%). qPCR showed that EDTA detached 99% of biofilm cells, and MSS detached 94% of biofilm cells (both P < .001). In contrast to EDTA, MSS was highly antimicrobial. The treatment promoted an ample log 7 reduction of the attached cells (P < .001), and almost no live cells were detected in the supernatant (P < .001). Positive correlations between CFU and qPCR with PMA were observed (r = 0.959 and r = 0.729). EDTA detached cells in biofilms with a minor antimicrobial effect. Besides a great antimicrobial effect, MSS also detached biofilm cells. These dispersals of biofilms give insights into new endodontic biofilm removal strategies. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.
Klupp, E-M; Both, A; Belmar Campos, C; Büttner, H; König, C; Christopeit, M; Christner, M; Aepfelbacher, M; Rohde, H
Vancomycin-resistant enterococci (VRE) are of ever-increasing importance, most notably in high-risk patient populations. Therapy options are often limited for these isolates, and apart from tigecycline and daptomycin, oxazolidinone linezolid is frequently administered. The broad usage of linezolid, however, has driven the emergence of linezolid-resistant VRE strains (LR-VRE), further shortening therapeutic options. Second-generation oxazolidinone tedizolid has the advantage of being active against a specific subset of LR-VRE, i.e. isolates expressing the plasmid-encoded chloramphenicol-florfenicol resistance (cfr) gene. Here we tested tedizolid activity in a collection of 30 LR Enterococcus faecium VRE (MIC range 32-256 mg/l) isolated between 2012 and 2015 from clinical and screening specimens. By pulsed field gel electrophoresis (PFGE) isolates were assigned to 16 clonal lineages. In three cases, linezolid-susceptible progenitor isolates of LR-VRE were isolated, thus demonstrating the de-novo emergence of the linezolid-resistant phenotype. PCR did not detect cfr, cfr(B) or novel oxazolidinone resistance gene optrA in LR-VRE. All isolates, however, carried mutations within the 23S rDNA. Compared to linezolid, tedizolid MICs were lower in all isolates (MIC range 2-32 mg/l), but remained above the FDA tedizolid breakpoint for E. faecalis at 0.5 mg/l. Thus, related to the predominant resistance mechanism, tedizolid is of limited value for treatment of most LR-VRE and represents a therapeutic option only for a limited subset of isolates.
Mesnage, Stéphane; Chau, Françoise; Dubost, Lionel; Arthur, Michel
Identification of the full complement of peptidoglycan hydrolases detected by zymogram in Enterococcus faecalis extracts led to the characterization of two novel hydrolases that we named AtlB and AtlC. Both enzymes have a similar modular organization comprising a central catalytic domain fused to two LysM peptidoglycan-binding modules. AtlB and AtlC displayed N-acetylmuramidase activity, as demonstrated by tandem mass spectrometry analyses of peptidoglycan fragments generated by the purified enzymes. The genes encoding AtlB and AtlC were deleted either alone or in combination with the gene encoding AtlA, a previously described N-acetylglucosaminidase. No autolytic activity was detected in the triple mutant indicating that AtlA, AtlB, and AtlC account for the major hydrolytic activities in E. faecalis. Analysis of cell size distribution by flow cytometry showed that deletion of atlA resulted in the formation of long chains. Thus, AtlA digests the septum and is required for cell separation after cell division. We found that AtlB could act as a surrogate for AtlA, although the enzyme was less efficient at septum digestion. Deletion of atlC had no impact on cell morphology. Labeling of the peptidoglycan with N-[14C]acetylglucosamine revealed an unusually slow turnover as compared with model organisms, almost completely dependent upon the combined activities of AtlA and AtlB. In contrast to atlA, the atlB and atlC genes are located in putative prophages. Because AtlB and AtlC were produced in the absence of cell lysis or production of phage progeny, these enzymes may have been hijacked by E. faecalis to contribute to peptidoglycan metabolism.
Eckert, Catherine; Lecerf, Maxime; Dubost, Lionel; Arthur, Michel; Mesnage, Stéphane
The major peptidoglycan hydrolase of Enterococcus faecalis, AtlA, has been identified, but its enzyme activity remains unknown. We have used tandem mass spectrometry analysis of peptidoglycan hydrolysis products obtained using the purified protein to show that AtlA is an N-acetylglucosaminidase. To gain insight into the regulation of its enzyme activity, the three domains of AtlA were purified alone or in combination following expression of truncated forms of the atlA gene in Escherichia coli or partial digestion of AtlA by proteinase K. The central domain of AtlA was catalytically active, but its activity was more than two orders of magnitude lower than that of the complete protein. Partial proteolysis of AtlA was detected in vivo: zymograms of E. faecalis extracts revealed two catalytically active protein bands of 62 and 72 kDa that were both absent in extracts from an atlA null mutant. Limited digestion of AtlA by proteinase K in vitro suggested that the proteolytic cleavage of AtlA in E. faecalis extracts corresponds to the truncation of the N-terminal domain, which is rich in threonine and glutamic acid residues. We show that the truncation of the N-terminal domain from recombinant AtlA has no impact on enzyme activity. The C-terminal domain of the protein, which contains six LysM modules bound to highly purified peptidoglycan, was required for optimal enzyme activity. These data indicate that AtlA is not produced as a proenzyme and that control of the AtlA glucosaminidase activity is likely to occur at the level of LysM-mediated binding to peptidoglycan.
Eckert, Catherine; Lecerf, Maxime; Dubost, Lionel; Arthur, Michel; Mesnage, Stéphane
The major peptidoglycan hydrolase of Enterococcus faecalis, AtlA, has been identified, but its enzyme activity remains unknown. We have used tandem mass spectrometry analysis of peptidoglycan hydrolysis products obtained using the purified protein to show that AtlA is an N-acetylglucosaminidase. To gain insight into the regulation of its enzyme activity, the three domains of AtlA were purified alone or in combination following expression of truncated forms of the atlA gene in Escherichia coli or partial digestion of AtlA by proteinase K. The central domain of AtlA was catalytically active, but its activity was more than two orders of magnitude lower than that of the complete protein. Partial proteolysis of AtlA was detected in vivo: zymograms of E. faecalis extracts revealed two catalytically active protein bands of 62 and 72 kDa that were both absent in extracts from an atlA null mutant. Limited digestion of AtlA by proteinase K in vitro suggested that the proteolytic cleavage of AtlA in E. faecalis extracts corresponds to the truncation of the N-terminal domain, which is rich in threonine and glutamic acid residues. We show that the truncation of the N-terminal domain from recombinant AtlA has no impact on enzyme activity. The C-terminal domain of the protein, which contains six LysM modules bound to highly purified peptidoglycan, was required for optimal enzyme activity. These data indicate that AtlA is not produced as a proenzyme and that control of the AtlA glucosaminidase activity is likely to occur at the level of LysM-mediated binding to peptidoglycan. PMID:17041059
Pankey, George A; Ashcraft, Deborah S
An emerging pathogen is Enterococcus faecium resistant to both linezolid and vancomycin (LRVRE). Antimicrobial combinations may be required for therapy and need to be evaluated. The combination of daptomycin and rifampin has demonstrated good in vitro activity against gram-positive bacteria, including E faecium. Telavancin, a newer lipoglycopeptide, has shown in vitro activity against E faecium. We evaluated the combination of telavancin and rifampin and compared the results to the combination of daptomycin and rifampin used previously on the same isolates. Twenty-four genetically unique (by pulsed-field gel electrophoresis), clinical LRVRE isolates were collected in the United States from 2001-2004. Etest minimal inhibitory concentrations (MICs) (μg/mL) were 0.064-8 for telavancin, 1-4 for daptomycin, and 0.012 to >32 for rifampin. In vitro synergy testing was performed in triplicate by an Etest MIC:MIC ratio method, and summation fractional inhibitory concentration (ΣFIC) was calculated: synergy ≤0.5; indifference >0.5-4; and antagonism >4. The Etest method showed synergy (ΣFICs of 0.1-0.5) with telavancin + rifampin in 20/24 (83%) isolates and indifference (ΣFICs of 0.6-0.8) in 4/24 (17%) isolates. Similarly, the daptomycin + rifampin combination showed synergy (ΣFICs of 0.1-0.5) in 21/24 (88%) isolates and indifference (ΣFICs of 0.6-1.0) in 3/24 (12%) isolates by the Etest method. No antagonism was found. In vitro synergy with both combinations (rifampin + telavancin or daptomycin) was 83% and 88%, respectively, by Etest against these LRVRE isolates. Although both daptomycin and telavancin in combination with rifampin showed a high incidence of synergistic activity, further in vitro synergy testing with this combination should be performed against additional E faecium isolates. In vitro synergy may or may not translate into in vivo effectiveness.
Full Text Available Introduction: Adequate biomechanical preparations, antibacterial irrigants, and intracanal medications to promote the elimination of bacteria and their products are required to succeed root canal treatment. Enterococcus faecalis with its biofilm is known as an important etiological agent in endodontic treatment failures. Chitosan, as a natural product, has an antibacterial activity and is considered less toxic to the periapical tissue than other irrigants. However, the use of this natural product needs to be examined to determine its effectiveness as a root canal irrigant in endodontic treatment; this can be done by comparing it with the most common endodontic irrigant (NaOCl 5.25% as a positive control. Objective: The objective of the study was to compare the effectiveness between 1–3% chitosan acetate (CA and 1–3% chitosan citrate (CC against E. faecalis biofilm formation after treatment for 15, 30, and 60 minutes. Methods: The study was conducted using 12 groups, including 1–3% CA, 1–3% CC, and control groups. E. faecalis biofilms in 96-well plates were exposed to each sample for 15, 30, or 60 minutes. Subsequently, the biofilms were stained with crystal violet solution, and the optical density value was measured using a microtiter plate reader at a wavelength of 600nm. Results: CA and CC were effective in eradicating E. faecalis biofilm. However, the levels of effectiveness of CC and CA depended on the concentration and application time. Three-way analysis of variance (ANOVA showed a significant difference between the irrigants (F = 5.92; p<0.05 and three application times (F = 6.50; p< 0.05. The CA was effective in eradicating biofilm after 15 minutes of application, whereas the CC was more effective after 30 and 60 minutes of application. Conclusion: CC and CA are both effective in eradicating E. faecalis biofilm.
Ogaki, Mayara Baptistucci; Rocha, Katia Real; Terra, MÁrcia Regina; Furlaneto, MÁrcia Cristina; Maia, Luciana Furlaneto
In the current study, a total of 135 enterococci strains from different sources were screened for the presence of the enterocin-encoding genes entA, entP, entB, entL50A, and entL50B. The enterocin genes were present at different frequencies, with entA occurring the most frequently, followed by entP and entB; entL50A and L50B were not detected. The occurrence of single enterocin genes was higher than the occurrence of multiple enterocin gene combinations. The 80 isolates that harbor at least one enterocin-encoding gene (denoted "Gene(+) strains") were screened for antimicrobial activity. A total of 82.5% of the Gene(+) strains inhibited at least one of the indicator strains, and the isolates harboring multiple enterocin-encoding genes inhibited a larger number of indicator strains than isolates harboring a single gene. The indicator strains that exhibited growth inhibition included Listeria innocua strain CLIP 12612 (ATCC BAA-680), Listeria monocytogenes strain CDC 4555, Enterococcus faecalis ATCC 29212, Staphylococcus aureus ATCC 25923, S. aureus ATCC 29213, S. aureus ATCC 6538, Salmonella enteritidis ATCC 13076, Salmonella typhimurium strain UK-1 (ATCC 68169), and Escherichia coli BAC 49LT ETEC. Inhibition due to either bacteriophage lysis or cytolysin activity was excluded. The growth inhibition of antilisterial Gene+ strains was further tested under different culture conditions. Among the culture media formulations, the MRS agar medium supplemented with 2% (w/v) yeast extract was the best solidified medium for enterocin production. Our findings extend the current knowledge of enterocin-producing enterococci, which may have potential applications as biopreservatives in the food industry due to their capability of controlling food spoilage pathogens.
Shen, Jiangchuan; Walsh, Brenna J C; Flores-Mireles, Ana Lidia; Peng, Hui; Zhang, Yifan; Zhang, Yixiang; Trinidad, Jonathan C; Hultgren, Scott J; Giedroc, David P
Recent studies of hydrogen sulfide (H 2 S) signaling implicate low molecular weight (LMW) thiol persulfides and other reactive sulfur species (RSS) as signaling effectors. Here, we show that a CstR protein from the human pathogen Enterococcus faecalis ( E. faecalis), previously identified in Staphylococcus aureus ( S. aureus), is an RSS-sensing repressor that transcriptionally regulates a cst-like operon in response to both exogenous sulfide stress and Angeli's salt, a precursor of nitroxyl (HNO). E. faecalis CstR reacts with coenzyme A persulfide (CoASSH) to form interprotomer disulfide and trisulfide bridges between C32 and C61', which negatively regulate DNA binding to a consensus CstR DNA operator. A Δ cstR strain exhibits deficiency in catheter colonization in a catheter-associated urinary tract infection (CAUTI) mouse model, suggesting sulfide regulation and homeostasis is critical for pathogenicity. Cellular polysulfide metabolite profiling of sodium sulfide-stressed E. faecalis confirms an increase in both inorganic polysulfides and LMW thiols and persulfides sensed by CstR. The cst-like operon encodes two authentic thiosulfate sulfurtransferases and an enzyme we characterize here as an NADH and FAD-dependent coenzyme A (CoA) persulfide reductase (CoAPR) that harbors an N-terminal CoA disulfide reductase (CDR) domain and a C-terminal rhodanese homology domain (RHD). Both cysteines in the CDR (C42) and RHD (C508) domains are required for CoAPR activity and complementation of a sulfide-induced growth phenotype of a S. aureus strain lacking cstB, encoding a nonheme Fe II persulfide dioxygenase. We propose that S. aureus CstB and E. faecalis CoAPR employ orthogonal chemistries to lower CoASSH that accumulates under conditions of cellular sulfide toxicity and signaling.
José Chibebe Junior
Full Text Available Enterococcus faecium has emerged as one of the most important pathogens in healthcare-associated infections worldwide due to its intrinsic and acquired resistance to many antibiotics, including vancomycin. Antimicrobial photodynamic therapy (aPDT is an alternative therapeutic platform that is currently under investigation for the control and treatment of infections. PDT is based on the use of photoactive dye molecules, widely known as photosensitizer (PS. PS, upon irradiation with visible light, produces reactive oxygen species that can destroy lipids and proteins causing cell death. We employed Galleria mellonella (the greater wax moth caterpillar fatally infected with E. faecium to develop an invertebrate host model system that can be used to study the antimicrobial PDT (alone or combined with antibiotics. In the establishment of infection by E. faecium in G. mellonella, we found that the G. mellonella death rate was dependent on the number of bacterial cells injected into the insect hemocoel and all E. faecium strains tested were capable of infecting and killing G. mellonella. Antibiotic treatment with ampicillin, gentamicin or the combination of ampicillin and gentamicin prolonged caterpillar survival infected by E. faecium (P = 0.0003, P = 0.0001 and P = 0.0001, respectively. In the study of antimicrobial PDT, we verified that methylene blue (MB injected into the insect followed by whole body illumination prolonged the caterpillar survival (P = 0.0192. Interestingly, combination therapy of larvae infected with vancomycin-resistant E. faecium, with antimicrobial PDT followed by vancomycin, significantly prolonged the survival of the caterpillars when compared to either antimicrobial PDT (P = 0.0095 or vancomycin treatment alone (P = 0.0025, suggesting that the aPDT made the vancomycin resistant E. faecium strain more susceptible to vancomycin action. In summary, G. mellonella provides an invertebrate model host to
Bardossy, Ana Cecilia; Alsafadi, Muhammad Yasser; Starr, Patricia; Chami, Eman; Pietsch, Jennifer; Moreno, Daniela; Johnson, Laura; Alangaden, George; Zervos, Marcus; Reyes, Katherine
There are limited controlled data demonstrating contact precautions (CPs) prevent methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus (VRE) infections in endemic settings. We evaluated changes in hospital-acquired MRSA and VRE infections after discontinuing CPs for these organisms. This is a retrospective study done at an 800-bed teaching hospital in urban Detroit. CPs for MRSA and VRE were discontinued hospital-wide in 2013. Data on MRSA and VRE catheter-associated urinary tract infections (CAUTIs), ventilator-associated pneumonia (VAP), central line-associated bloodstream infections (CLABSIs), surgical site infections (SSIs), and hospital-acquired MRSA bacteremia (HA-MRSAB) rates were compared before and after CPs discontinuation. There were 36,907 and 40,439 patients hospitalized during the two 12-month periods: CPs and no CPs. Infection rates in the CPs and no-CPs periods were as follows: (1) MRSA infections: VAP, 0.13 versus 0.11 (P = .84); CLABSI, 0.11 versus 0.19 (P = .45); SSI, 0 versus 0.14 (P = .50); and CAUTI, 0.025 versus 0.033 (P = .84); (2) VRE infections: CAUTI, 0.27 versus 0.13 (P = .19) and CLABSI, 0.29 versus 0.3 (P = .94); and (3) HA-MRSAB rates: 0.14 versus 0.11 (P = .55), respectively. Discontinuation of CPs did not adversely impact endemic MRSA and VRE infection rates. Copyright © 2017 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.
Nallapareddy, Sreedhar R; Singh, Kavindra V; Murray, Barbara E
Enterococcus faecium is a multidrug-resistant opportunist causing difficult-to-treat nosocomial infections, including endocarditis, but there are no reports experimentally demonstrating E. faecium virulence determinants. Our previous studies showed that some clinical E. faecium isolates produce a cell wall-anchored collagen adhesin, Acm, and that an isogenic acm deletion mutant of the endocarditis-derived strain TX0082 lost collagen adherence. In this study, we show with a rat endocarditis model that TX0082 Deltaacm::cat is highly attenuated versus wild-type TX0082, both in established (72 h) vegetations (P Acm the first factor shown to be important for E. faecium pathogenesis. In contrast, no mortality differences were observed in a mouse peritonitis model. While 5 of 17 endocarditis isolates were Acm nonproducers and failed to adhere to collagen in vitro, all had an intact, highly conserved acm locus. Highly reduced acm mRNA levels (>or=50-fold reduction relative to an Acm producer) were found in three of these five nonadherent isolates, including the sequenced strain TX0016, by quantitative reverse transcription-PCR, indicating that acm transcription is downregulated in vitro in these isolates. However, examination of TX0016 cells obtained directly from infected rat vegetations by flow cytometry showed that Acm was present on 40% of cells grown during infection. Finally, we demonstrated a significant reduction in E. faecium collagen adherence by affinity-purified anti-Acm antibodies from E. faecium endocarditis patient sera, suggesting that Acm may be a potential immunotarget for strategies to control this emerging pathogen.
Full Text Available Cell-cell communication mediated by peptide pheromones (cCF10 [CF] is essential for high-frequency plasmid transfer in vitro in Enterococcus faecalis. To examine the role of pheromone signaling in vivo, we established either a CF-producing (CF+ recipient or a recipient producing a biologically inactive variant of CF (CF− recipient in a germfree mouse model 3 days before donor inoculation and determined transfer frequencies of the pheromone-inducible plasmid pCF10. Plasmid transfer was detected in the upper and middle sections of the intestinal tract 5 h after donor inoculation and was highly efficient in the absence of antibiotic selection. The transconjugant/donor ratio reached a maximum level approaching 1 on day 4 in the upper intestinal tract. Plasmid transfer was significantly lower with the CF− recipient. While rescue of the CF− mating defect by coculture with CF+ recipients is easily accomplished in vitro, no extracellular complementation occurred in vivo. This suggests that most pheromone signaling in the gut occurs between recipient and donor cells in very close proximity. Plasmid-bearing cells (donors plus transconjugants steadily increased in the population from 0.1% after donor inoculation to about 10% at the conclusion of the experiments. This suggests a selective advantage of pCF10 carriage distinct from antibiotic resistance or bacteriocin production. Our results demonstrate that pheromone signaling is required for efficient pCF10 transfer in vivo. In the absence of CF+ recipients, a low level of transfer to CF− recipients occurred in the gut. This may result from low-level host-mediated induction of the donors in the gastrointestinal (GI tract, similar to that previously observed in serum.
Full Text Available Introduction: The ultimate goal of endodontic treatment is to eliminate the bacterial infection in the root canal system. While mechanical debridement combined with chemical irrigation removes the bulk of microorganisms, residual bacteria are readily detectable in approximately one-half of teeth just prior to obturation. Laser light can be used to destroy bacteria. This in vitro study was performed to evaluate the effect of diode laser and calcium hydroxide on mono-infected dental canals.Methods: Fifty five single-rooted human premolars were prepared and contaminated with Enterococcus faecalis. After three weeks of incubation, the samples were divided into three experimental groups (n = 15 and two control groups (n = 5. In the first and second groups, the teeth were rinsed for 5 min with either sterile saline or 5.25% NaOCl and irradiated with a 810-nm diode laser at 1.5 W output for 5 × 4s. In the third group, the teeth were rinsed with 5.25% NaOCl and then Ca(OH2 paste was inserted in the canals for 1 week. Intracanal bacterial sampling was done and the samples were plated to determine the CFU count. Results: 5.25% NaOCl plus laser was as effective as calcium hydroxide and significantly more effective than sterile saline (P>0.05 in elimination of E. faecalis. Complete elimination of E. faecalis was seen only for the one week calcium hydroxide treatment. Conclusion: Combination therapy with NaOCl irrigation and diode laser irradiation can be recommended as an effective treatment option for elimination of E. faecalis from the root canal system.
Kiran Rahul Halkai
Full Text Available Background: The main objective of endodontic therapy is complete elimination and prevention of bacteria from the root canal system; however, it is difficult due to anatomical ramifications of root canal system and growing resistant microbes to available disinfectants. Therefore, to overcome this problem, newer antimicrobial agents have to be developed. Aims: The aim of the study was to evaluate the antibacterial efficacy of fungal-derived biosynthesized silver nanoparticles (AgNPs against Enterococcus faecalis. Materials and Methods: Freshly prepared silver nanoparticles using the endophytic fungi Fusarium semitectum, characterized by different techniques were used to evaluate the antibacterial efficacy against E. faecalis by agar well diffusion method measuring the zone of inhibition using different concentrations of nanoparticles (AgNPs (A [20 μl], B [40 μl], C [60 μl], D [80 μl], and E [100 μl], F (0.2% chlorhexidine [CHX], G (2% CHX, H (ampicillin, and I (distilled water were used as control groups. Statistical Analysis Used: One-way ANOVA followed by post hoc Tukey multiple comparison test was done. Results: AgNPs (100 ml showed highest zone of inhibition 19.5 mm against E. faecalis. CHX (0.2% 14.52 mm, CHX (2% 20.02 mm, and ampicillin showed highest mean zone of inhibition 20.5 mm and distilled water showed no zone of inhibition. Results indicate no significant difference between E (100 μl, G (2% CHX, and H (ampicillin (P < 0.0001. Conclusions: Biosynthesized AgNPs exhibit efficient antibacterial activity against E. faecalis and therefore can be used as root canal irrigant or intracanal medicament for root canal disinfection.
Abeer A Al-Mahdi
Full Text Available Aim: The aim of this investigation was to compare the efficacy of passive ultrasonic irrigation (PUI with either 2.5% sodium hypochlorite (NaOCl or saline, with that of conventional syringe irrigation on intraradicular Enterococcus faecalis biofilm. Materials and Methods: Biofilms of E. faecalis were established over 21 days in 80 single roots that had undergone biomechanical preparation followed by gamma radiation. Biofilms were treated for 1 min with 2.5% NaOCl/PUI (Group 1, 2.5% NaOCl (Group 2, sterile saline/PUI (Group 3, and sterile saline (Group 4. The positive control (n = 4 was used to confirm the presence of biofilm before various treatments. Additional four samples that served as a negative control were used to confirm the sterility of the samples. Biofilm eradication was evaluated by Colony Forming Unit (CFU quantification and scanning electron microscopy (SEM. Results: The median of CFUs of S1 was significantly higher than that of S2 in all experimental groups. SEM examination showed a significant difference between the positive control and the experimental groups (P < 0.001, with the highest score of biofilm in the positive control group followed by Group 4 and both groups were not statistically significant from each other (P = 0.067. Following various treatments, the highest scores of biofilm were observed in the coronal third and the least were in the apical third. Conclusions: PUI did not increase the effectiveness of NaOCl irrigation on biofilm removal, however, PUI enhanced biofilm disturbance when used with saline. The least mean score of remaining biofilm was in the apical third of all treatment groups compared to other thirds.
Teik Hwa Ong
Full Text Available Propolis obtained from bee hives is a natural substance with antimicrobial properties. It is limited by its insolubility in aqueous solutions; hence ethanol and ethyl acetate extracts of Malaysian propolis were prepared. Both the extracts displayed antimicrobial and anti-biofilm properties against Enterococcus faecalis, a common bacterium associated with hospital-acquired infections. High performance liquid chromatography (HPLC analysis of propolis revealed the presence of flavonoids like kaempferol and pinocembrin. This study investigated the role of propolis developed into nanoparticles with chitosan for its antimicrobial and anti-biofilm properties against E. faecalis. Bacteria that grow in a slimy layer of biofilm are resistant to penetration by antibacterial agents. The use of nanoparticles in medicine has received attention recently due to better bioavailability, enhanced penetrative capacity and improved efficacy. A chitosan-propolis nanoformulation was chosen based on ideal physicochemical properties such as particle size, zeta potential, polydispersity index, encapsulation efficiency and the rate of release of the active ingredients. This formulation inhibited E. faecalis biofilm formation and reduced the number of bacteria in the biofilm by ~90% at 200 μg/ml concentration. When tested on pre-formed biofilms, the formulation reduced bacterial number in the biofilm by ~40% and ~75% at 200 and 300 μg/ml, respectively. The formulation not only reduced bacterial numbers, but also physically disrupted the biofilm structure as observed by scanning electron microscopy. Treatment of biofilms with chitosan-propolis nanoparticles altered the expression of biofilm-associated genes in E. faecalis. The results of this study revealed that chitosan-propolis nanoformulation can be deemed as a potential anti-biofilm agent in resisting infections involving biofilm formation like chronic wounds and surgical site infections.
Chang, Ping Chin; Li, Hua Yu; Tang, Hung Jen; Liu, Jien Wei; Wang, Jhi Joung; Chuang, Yin Ching
Little is known about the possible synergism of baicalein, a bioactive flavone of Scutellariae radix (a Chinese herb), when used in conjunction with other antimicrobial agents against vancomycin-resistant Enterococcus (VRE). This in vitro study examined the possible synergism of the combination of baicalein and gentamicin against VRE. Minimal inhibitory concentrations (MICs) of baicalein as well as gentamicin were determined against 39 clinical isolates of VRE by the agar dilution method. Synergistic activities were determined using the checkerboard method based on the fractional inhibitory concentration indices and also the time-kill method. Further time-kill studies were conducted with these two agents against one randomly chosen clinical isolate, VRE-096. Minimal concentrations inhibiting 50% (MIC(50)) and 90% (MIC(90)) of isolates for baicalein and gentamicin were all >256 microg/mL. Synergism between baicalein and gentamicin was demonstrated against four clinical isolates of VRE (VRE-70, VRE-940, VRE-096 and VRE-721). When approximately 5 x 10(5) colony-forming units/mL of VRE-096 was incubated with both baicalein at a concentration of 32 microg/mL (1/8 x MIC) and gentamicin at a concentration of 128 microg/mL (1/2 x MIC), there was an inhibitory effect against VRE that persisted for 48 h. At 48 h, the combination of baicalein and gentamicin at these respective concentrations resulted in a reduction of growth by approximately 2 orders of magnitude compared to that for the starting inoculum and by 3 orders of magnitude compared to that for baicalein alone, the more active single agent. This study demonstrated that baicalein and gentamicin can act synergistically in inhibiting VRE in vitro.
... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3355 Listeria spp... from clinical specimens. Additionally, some of these reagents consist of Listeria spp. antisera... clinical specimens. The identification aids in the diagnosis of listeriosis, a disease caused by bacteria...
Schwaiger, Karin; Bauer, Johann; Hörmansdorfer, Stefan; Mölle, Gabriele; Preikschat, Petra; Kämpf, Peter; Bauer-Unkauf, Ilse; Bischoff, Meike; Hölzel, Christina
Ampicillin and vancomycin are important antibiotics for the therapy of Enterococcus faecalis infections. The ampicillin resistance gene pbp5 is intrinsic in Enterococcus faecium. The vanC1 gene confers resistance to vancomycin and serves as a species marker for Enterococcus gallinarum. Both genes are chromosomally located. Resistance to ampicillin and vancomycin was determined in 484 E. faecalis of human and porcine origin by microdilution. Since E. faecalis are highly skilled to acquire resistance genes, all strains were investigated for the presence of pbp5 (and, in positive strains, for the penicillin-binding protein synthesis repressor gene psr) and vanC1 (and, in positive strains, for vanXYc and vanT) by using polymerase chain reaction (PCR). One porcine and one human isolate were phenotypically resistant to ampicillin; no strain was vancomycin resistant. Four E. faecalis (3/1 of porcine/human origin) carried pbp5 (MIC=1 mg/L), and four porcine strains were vanC1 positive (minimum inhibitory concentration [MIC]=1 mg/L). Real-time reverse transcriptase (RT)-PCR revealed that the genes were not expressed. The psr gene was absent in the four pbp5-positive strains; the vanXYc gene was absent in the four vanC1-positive strains. However, vanT of the vanC gene cluster was detected in two vanC1-positive strains. To our knowledge, this is the first report on the presence of pbp5, identical with the "E. faecium pbp5 gene," and of vanC1/vanT in E. faecalis. Even if resistance is not expressed in these strains, this study shows that E. faecalis have a strong ability to acquire resistance genes-and potentially to spread them to other bacteria. Therefore, close monitoring of this species should be continued.
Leila Shafiei Bafti
Full Text Available BACKGROUND AND AIM: Due to their antimicrobial and dental plaque control activity, mouthwashes lead to an improvement in oral health. Although chemical mouthwashes have demonstrated the greatest antimicrobial and anti-inflammatory effects, their usage has been limited because of their numerous side effects. This study was conducted in vivo to determine the antibacterial and antifungal effects of Persica herbal mouthwash containing Salvadore persica, mint, and yarrow in comparison with a placebo. METHODS: In this experimental, single-blind study, 80 dentistry students, who were eager to participate in the study, were randomly allocated into two groups of forty. One group was given Persica while the other group received a placebo. They were asked to apply the mouthwash twice a day for four weeks. The participants were unaware of the mouthwash type. Saliva sampling was conducted in all cases before and after mouthwash application and the samples were dispatched to a laboratory for microbial culture (Sabouraud and Clark-Kenner media culture. After two weeks, the washout times in groups were swapped with each other and the sampling was conducted just like before. Finally, the data were analyzed using independent and paired t-test. P values of less than 0.05 were considered significant. RESULTS: The mean age of participants was 23.20 ± 4.14. Persica mouthwash significantly decreased the count of Candida albicans (1.43 ± 0.15 to 0.8 ± 0.35 (P < 0.001 and Enterococcus faecalis (0.93 ± 1.76 to 0.71 ± 2.10 (P = 0.008. CONCLUSION: The statistical tests revealed that applying Persica mouthwash would result in a significant decrease in the Candida albicans and Enterococcus faecalis count. Regarding the significant and desirable effect of Persica on Candida albicans and Enterococcus faecalis, and its herbal origin, its application could be suggested to reduce oral microbes and infections from fungi and enterococci.
Kumar, Manoj; Srivastava, Sheela
Enterococci are well-known producers of antimicrobial peptides (enterocins) that possess potential as biopreservatives in food. In this study, divalent cations and release of intracellular potassium were used to assess the mechanism of interaction and killing of enterocin LR/6 produced by Enterococcus faecium LR/6 on three target Gram-positive and Gram-negative bacteria, namely Micrococcus luteus, Enterococcus sp. strain LR/3 and Escherichia coli K-12. Whilst treatment with enterocin LR/6 in all cases led to a significant loss of viability, suggesting a bactericidal mode of action, E. coli K-12 showed better tolerance than the other two strains. Bacteriocins have generally been reported to create pores in the membrane of sensitive cells and this function is diminished by divalent cations. In this study it was shown that Ca(2+) and Mg(2+) markedly improved the viability of enterocin LR/6-treated cells in a concentration-dependent manner. K(+) release as a sign of membrane leakiness was higher in M. luteus compared with the other two test strains. In agreement with the viability response, pre-exposure to Ca(2+) and Mg(2+) substantially reduced the amount of K(+) leakage by M. luteus and Enterococcus sp.; in the case of E. coli K-12, no leakage of K(+) was recorded. These results suggest that enterocin LR/6, which possesses good antibacterial potential, may not be very effective as a preservative in foods containing high concentrations of calcium and magnesium. Copyright © 2011 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.
Full Text Available Background: Enterococci have emerged as a major cause of nosocomial infections and within this group, Enterococcus faecalis and Enterococcus faecium cause the majority of human and livestock enterococcal infections. In this article, we tried to determine antibiotics and metals resistance patterns of E. faecalis and E. faecium strains. Methods: One hundred sixty different strains of E. faecalis and E. faecium were collected from livestock sewage and the human fecal waste during 15 months. Then bacterial antibiotics sensitivity tests were carried out using the Agar disc diffusion method. Results: Generally, 100% of E. faecalis strains separated from human and livestock sources (i.e. sheep showed penicillin (P/ kanamycin (K/ nitrofurantoin (N/ loracarbef (L/ Ciprofloxacin (Cc/ ampicillin (AN/ nalidixic acid (NA/ sulfamethoxazole (S antibiotics resistance patterns. In addition, 55% of isolated E. faecium showed P/S/AN/NA antibiotics resistance patterns. Each strain showed a resistance to at least two aminoglycoside antibiotics. However, E. faecalis strains from human and the livestock sources showed 94% and 100% of resistance to nitrofurantoin, respectively. The effects of different metal concentrations was evaluated in both strains. The agar dilution method was applied in this stage. Hg at 0.05 mmol/L of minimum inhibitory concentration (MIC showed toxicity to both the human and livestock Enterococcus strains. Cadmium at 1 mmol/L and 0.5 mmol/L concentrations had the most toxicity to E. faecalis and E. faecium strains, respectively. Obviously, toxicity to bacteria is less than other metals. As a result, Zn/Ni/Cu/Co resistance pattern is suggested for both strains. Finally, antibiotics and heavy metals resistance patterns were monitored simultaneously. Conclusion: Almost all E. faecalis strains isolated from humans and livestock showed antibiotics and heavy metals resistance patterns of P/K/L/Cc/S/AN/NA/Zn/Cu/Co simultaneously. Moreover, 55% of E
Gaaloul, N; ben Braiek, O; Hani, K; Volski, A; Chikindas, M L; Ghrairi, T
To assess the antimicrobial properties of lactic acid bacteria from Tunisian raw bovine milk. A bacteriocin-producing Enterococcus faecium strain was isolated from raw cow milk with activity against Gram-positive and Gram-negative bacteria. Antimicrobial substances produced by this strain were sensitive to proteolytic enzymes and were thermostable and resistant to a broad range of pH (2-10). Mode of action of antimicrobial substances was determined as bactericidal. Maximum activity was reached at the end of the exponential growth phase when checked against Listeria ivanovii BUG 496 (2366.62 AU ml(-1)). However, maximum antimicrobial activity against Pseudomonas aeruginosa 28753 was recorded at the beginning of the exponential growth phase. Enterococcus faecium GGN7 was characterized as free from virulence factors and was susceptible to tested antibiotics. PCR analysis of the micro-organism's genome revealed the presence of genes coding for enterocins A and B. Mass spectrometry analysis of RP-HPLC active fractions showed molecular masses corresponding to enterocins A (4835.77 Da) and B (5471.56 Da), and a peptide with a molecular mass of 3215.5 Da active only against Gram-negative indicator strains. The latter was unique in the databases. Enterococcus faecium GGN7 produces three bacteriocins with different inhibitory spectra. Based on its antimicrobial properties and safety, Ent. faecium GGN7 is potentially useful for food biopreservation. The results suggest the bacteriocins from GGN7 strain could be useful for food biopreservation. © 2014 The Society for Applied Microbiology.
Nomura, Takahiro; Tanimoto, Koichi; Shibayama, Keigo; Arakawa, Yoshichika; Fujimoto, Shuhei; Ike, Yasuyoshi; Tomita, Haruyoshi
Five VanN-type vancomycin-resistant Enterococcus faecium strains were isolated from a sample of domestic chicken meat in Japan. All isolates showed low-level resistance to vancomycin (MIC, 12 mg/liter) and had the same pulsed-field gel electrophoresis profile. The vancomycin resistance was encoded on a large plasmid (160 kbp) and was expressed constitutively. The VanN-type resistance operon was identical to the first resistance operon to be reported, with the exception of a 1-bp deletion in vanT(N) and a 1-bp substitution in vanS(N).
Radhouani, Hajer; Poeta, Patrícia; Pinto, Luís; Miranda, Júlio; Coelho, Céline; Carvalho, Carlos; Rodrigues, Jorge; López, María; Torres, Carmen; Vitorino, Rui; Domingues, Pedro; Igrejas, Gilberto
Enterococci have emerged as the third most common cause of nosocomial infections, requiring bactericidal antimicrobial therapy. Although vancomycin resistance is a major problem in clinics and has emerged in an important extend in farm animals, few studies have examined it in wild animals. To determine the prevalence of vanA-containing Enterococcus strains among faecal samples of Seagulls (Larus cachinnans) of Berlengas Natural Reserve of Portugal, we developed a proteomic approach integrated with genomic data. The purpose was to detect the maximum number of proteins that vary in different enterococci species which are thought to be connected in some, as yet unknown, way to antibiotic resistance. From the 57 seagull samples, 54 faecal samples showed the presence of Enterococcus isolates (94.7%). For the enterococci, E. faecium was the most prevalent species in seagulls (50%), followed by E. faecalis and E. durans (10.4%), and E. hirae (6.3%). VanA-containing enterococcal strains were detected in 10.5% of the 57 seagull faecal samples studied. Four of the vanA-containing enterococci were identified as E. faecium and two as E. durans. The tet(M) gene was found in all five tetracycline-resistant vanA strains. The erm(B) gene was demonstrated in all six erythromycin-resistant vanA strains. The hyl virulence gene was detected in all four vanA-containing E. faecium isolates in this study, and two of them harboured the purK1 allele. In addition these strains also showed ampicillin and ciprofoxacin resistance. The whole-cell proteomic profile of vanA-containing Enterococcus strains was applied to evaluate the discriminatory power of this technique for their identification. The major differences among species-specific profiles were found in the positions corresponding to 97-45 kDa. Sixty individualized protein spots for each vanA isolate was identified and suitable for peptide mass fingerprinting measures by spectrometry measuring (MALDI/TOF MS) and their identification
Full Text Available Abstract Background Enterococci have emerged as the third most common cause of nosocomial infections, requiring bactericidal antimicrobial therapy. Although vancomycin resistance is a major problem in clinics and has emerged in an important extend in farm animals, few studies have examined it in wild animals. To determine the prevalence of vanA-containing Enterococcus strains among faecal samples of Seagulls (Larus cachinnans of Berlengas Natural Reserve of Portugal, we developed a proteomic approach integrated with genomic data. The purpose was to detect the maximum number of proteins that vary in different enterococci species which are thought to be connected in some, as yet unknown, way to antibiotic resistance. Results From the 57 seagull samples, 54 faecal samples showed the presence of Enterococcus isolates (94.7%. For the enterococci, E. faecium was the most prevalent species in seagulls (50%, followed by E. faecalis and E. durans (10.4%, and E. hirae (6.3%. VanA-containing enterococcal strains were detected in 10.5% of the 57 seagull faecal samples studied. Four of the vanA-containing enterococci were identified as E. faecium and two as E. durans. The tet(M gene was found in all five tetracycline-resistant vanA strains. The erm(B gene was demonstrated in all six erythromycin-resistant vanA strains. The hyl virulence gene was detected in all four vanA-containing E. faecium isolates in this study, and two of them harboured the purK1 allele. In addition these strains also showed ampicillin and ciprofoxacin resistance. The whole-cell proteomic profile of vanA-containing Enterococcus strains was applied to evaluate the discriminatory power of this technique for their identification. The major differences among species-specific profiles were found in the positions corresponding to 97-45 kDa. Sixty individualized protein spots for each vanA isolate was identified and suitable for peptide mass fingerprinting measures by spectrometry measuring
Nallapareddy, Sreedhar R; Sillanpää, Jouko; Ganesh, Vannakambadi K; Höök, Magnus; Murray, Barbara E
Strains of Enterococcus faecium express a cell wall-anchored protein, Acm, which mediates adherence to collagen. Here, we (i) identify the minimal and high-affinity binding subsegments of Acm and (ii) show that anti-Acm immunoglobulin Gs (IgGs) purified against these subsegments reduced E. faecium TX2535 strain collagen adherence up to 73 and 50%, respectively, significantly more than the total IgGs against the full-length Acm A domain (28%) (P Acm adherence with functional subsegment-specific antibodies raises the possibility of their use as therapeutic or prophylactic agents.
Caruso, Gabriella; Monticelli, L S; Caruso, R; Bergamasco, A
A direct, microscopic fluorescent antibody method was developed to detect the occurrence of Enterococcus faecium in coastal aquatic environments and was compared with the conventional membrane filtering method. The "in situ" application of the antibody-based protocol in the analysis of water samples collected from coastal polyhaline habitats demonstrated good sensitivity and ease of implementation. Data obtained with the microscopic technique were in agreement with those obtained from culture counts. The fluorescent antibody method proved to be a rapid and reliable technique for the detection of E. faecium. The advantages and limitations intrinsic to the method are discussed, highlighting the potential of this new technique for monitoring coastal aquatic environments.
Nishi, Letícia; Bergamasco, Rosângela; Toledo, Max Jean de Ornelas; Falavigna, Dina Lúcia Morais; Gomes, Mônica Lúcia; Mota, Lúcio Tadeu; Falavigna-Guilherme, Ana Lúcia
The objective of this study was to investigate the occurrence of cysts of Giardia spp. and oocysts of Cryptosporidium spp. in waters of the Ivaí Indigenous Land, Brazil. Samples of river and spring water and of treated water were filtered and analyzed by direct immunofluorescence (Merifluor kit, Meridian Bioscience, Cincinnati, Ohio). Of 21 samples, 7 from each locality, 3 (3/7, 42.8%) from a river were positive for Giardia (mean concentration 2.57 cysts/L), and 1 (1/7, 14.3%) was positive for Cryptosporidium (6 oocysts/L). From springs, 1 sample (1/7, 14.3%) was positive for Cryptosporidium (6 oocysts/L). One sample (1/7, 14.3%) from treated water was positive for both, with 4 oocysts/L and 2 cysts/L. Giardia was the more frequent protozoan present.
Beyhan, Yunus Emre; Mungan, Mesut; Babür, Cahit
Ticks are a threat to human health by blood sucking and vectoring many disease agents. Tick-borne diseases are seen all over the world and play an important role in the dissemination of diseases. Although many of the tick species are present in Turkey, Amblyomma genus is seen more in South America and Africa. In this case,a person returning to the country after travelling to Africa who presented to the hospital complaining of tick bites and brought ticks to the parasitology laboratory was identified as Ablyomma spp. nymph. This case is a report concerning care of tick bites when travelling abroad and shows that these ticks can transmit disease agents from abroad.
Full Text Available Urinary tract infections (UTIs in women are a growing clinical concern. The most frequent risk factors of UTIs with fungal aetiology in women are: antibiotic therapy (especially broad-spectrum antibiotics, immunosuppressive therapy, diabetes, malnutrition, pregnancy, and frequent intercourse. The aim of the study was to analyse urinary tract infections with Candida spp. aetiology in women hospitalised at the Clinical Hospital in Poznań, Poland, between 2009 and 2011. The investigations revealed that as many as 71% of positive urine cultures with Candida fungi came from women. The following fungi were most frequently isolated from the patients under analysis: C. albicans (47%, C. glabrata (31%, C. tropicalis (6%, C. krusei (3%. In order to diagnose a UTI the diagnosis cannot be based on a single result of a urine culture. Due to the small number of antifungal drugs and high costs of treatment, antifungal drugs should be applied with due consideration and care.
Lafri, Ismail; El Hamzaoui, Basma; Bitam, Idir; Leulmi, Hamza; Lalout, Reda; Mediannikov, Oleg; Chergui, Mohamed; Karakellah, Mohamed; Raoult, Didier; Parola, Philippe
Argasid ticks (soft ticks) are blood-feeding arthropods that can parasitize rodents, birds, humans, livestock and companion animals. Ticks of the Ornithodoros genus are known to be vectors of relapsing fever borreliosis in humans. In Algeria, little is known about relapsing fever borreliosis and other bacterial pathogens transmitted by argasid ticks. Between May 2013 and October 2015, we investigated the presence of soft ticks in 20 rodent burrows, 10 yellow-legged gull (Larus michahellis) nests and animal shelters in six locations in two different bioclimatic zones in Algeria. Six species of argasid ticks were identified morphologically and through 16S rRNA gene sequencing. The presence and prevalence of Borrelia spp., Bartonella spp., Rickettsia spp. and Anaplasmataceae was assessed by qPCR template assays in each specimen. All qPCR-positive samples were confirmed by standard PCR, followed by sequencing the amplified fragments. Two Borrelia species were identified: Borrelia hispanica in Ornithodoros occidentalis in Mostaganem, and Borrelia cf. turicatae in Carios capensis in Algiers. One new Bartonella genotype and one new Anaplasmataceae genotype were also identified in Argas persicus. The present study highlights the presence of relapsing fever borreliosis agents, although this disease is rarely diagnosed in Algeria. Other bacteria of unknown pathogenicity detected in argasid ticks which may bite humans deserve further investigation.
Full Text Available Argasid ticks (soft ticks are blood-feeding arthropods that can parasitize rodents, birds, humans, livestock and companion animals. Ticks of the Ornithodoros genus are known to be vectors of relapsing fever borreliosis in humans. In Algeria, little is known about relapsing fever borreliosis and other bacterial pathogens transmitted by argasid ticks.Between May 2013 and October 2015, we investigated the presence of soft ticks in 20 rodent burrows, 10 yellow-legged gull (Larus michahellis nests and animal shelters in six locations in two different bioclimatic zones in Algeria. Six species of argasid ticks were identified morphologically and through 16S rRNA gene sequencing. The presence and prevalence of Borrelia spp., Bartonella spp., Rickettsia spp. and Anaplasmataceae was assessed by qPCR template assays in each specimen. All qPCR-positive samples were confirmed by standard PCR, followed by sequencing the amplified fragments. Two Borrelia species were identified: Borrelia hispanica in Ornithodoros occidentalis in Mostaganem, and Borrelia cf. turicatae in Carios capensis in Algiers. One new Bartonella genotype and one new Anaplasmataceae genotype were also identified in Argas persicus.The present study highlights the presence of relapsing fever borreliosis agents, although this disease is rarely diagnosed in Algeria. Other bacteria of unknown pathogenicity detected in argasid ticks which may bite humans deserve further investigation.
Nahar, Kamrun; Goyer, Claudia; Zebarth, Bernie J; Burton, David L; Whitney, Sean
Potato cultivars vary in their tolerance to common scab (CS), however how they affect CS-causing Streptomyces spp. populations over time is poorly understood. This study investigated the effects of potato cultivar on pathogenic Streptomyces spp. abundance, measured using quantitative PCR, in three spatial locations in a CS-infested field: 1) soil close to the plant (SCP); 2) rhizosphere (RS); and 3) geocaulosphere (GS) soils. Two tolerant (Gold Rush, Hindenburg) and two susceptible cultivars (Green Mountain, Agria) were tested. The abundance of pathogenic Streptomyces spp. significantly increased in late August compared with other dates in RS of susceptible cultivars in both years. Abundance of pathogenic Streptomyces spp., when averaged over locations and time, was significantly greater in susceptible cultivars compared with tolerant cultivars in 2014. Principal coordinates analysis showed that SCP and RS soil properties (pH, organic carbon and nitrogen concentrations) explained 68% and 76% of total variation in Streptomyces spp. abundance among cultivars in 2013, respectively, suggesting that cultivars influenced CS pathogen growth conditions. The results suggested that the genetic background of potato cultivars influenced the abundance of pathogenic Streptomyces spp., with 5 to 6 times more abundant Streptomyces spp. in RS of susceptible cultivars compared with tolerant cultivars, which would result in substantially more inoculum left in the field after harvest. .
Aguilar Galvez A.
Full Text Available The aim of this work was to isolate and to characterize strains of lactic acid bacteria (LAB with bacteriocin-like inhibitory activity from 27 traditional cheeses artisanal-produced obtained from different Peruvian regions. Twenty Gram+ and catalasenegative strains among 2,277 isolates exhibited bacteriocin-like inhibitory activity against Listeria monocytogenes CWBIB2232 as target strain. No change in inhibitory activity was observed after organic acid neutralization and treatment with catalase of the cell-free supernatant (CFS. The proteinic nature of the antimicrobial activity was confirmed for the twenty LAB strains by proteolytic digestion of the CFS. Two strains, CWBI-B1431 and CWBI-B1430, with the best antimicrobial activity were selected for further researches. These strains were taxonomically identified by phenotypic and genotypic analyses as Enterococcus mundtii (CWBI-B1431 and Enterococcus faecium (CWBI-B1430. The two strains were sensitive to vancomycin (MIC 2 μg.ml-1 and showed absence of haemolysis.
N. V. Gonchar
Full Text Available Introduction. Possibilities of using probiotic enterococci in premature neonates undergoing inpatient antibacterial therapy remains understudied. The article is aimed at analyzing clinical and pharmacoeconomic reasonability of using probiotic Enterococcus faecium L3 strain in premature infants with very low body weight in the framework of complex inpatient developmental care. Patients and methods. 55 children randomized into 2 groups were observed: the control group (n = 26 was undergoing standard developmental care program, the primary group (n = 29 was introduced liquid probiotic Enterococcus faecium L3 strain (titer — 108 CFU/ml or more (0.5 ml TID for 14 days after attaining the enteral feeding volume of 5.0 ml. Results. Analysis of the clinical symptoms characteristic of non-smooth course of developmental care over premature infants helped to reveal higher frequency of infectious complications in the control group children than in the primary group (14 [53.8%] vs. 6 [20.7%]; p < 0.05. Acute food intolerance was observed less frequently in the primary group than in the control group (6 [20.7%] vs. 10 [38.5%], p > 0.05. The primary group's children featured significant decrease in the frequency of monocytosis, positive changes of intestinal microbiotic composition (increase in the amount of bifidum bacteria, lactobacilli, enterococci, decrease in the amount of Clostridium difficile and antibiotic-resistant clinical Klebsiella pneumoniae strains. Conclusion. Favorable outcome of developmental care over premature infants (absence of infectious complications was less expensive in the primary group's children.
Full Text Available he aim of this study was to determine the prevalence and antibiotic resistance of enterococcii and E. coli strains isolated from dairy calves and lambs. Susceptibilities of isolated enterococci were tested using the disk diffusion method. The interpretation of inhibition zones around the disks was according to CLSI 2004 Performance standards for antimicrobial susceptibility testing. In our study, all isolates (E. coli and enterococci were multiresistant (100% to tetracycline, streptomycin and compound sulphonamides. Lower levels of resistance to enrofloxacin were noted. Antimicrobial resistance profiles of Enterococcus sp. isolated from lambs indicated that the highest percentage of susceptibility was exhibited to tetracycline (100% and streptomycin (100% and compound sulphonamides (100%. The intermediate resistance was exhibited against compound enrofloxacin (80%. The high frequencies of resistant isolates of Enterococcus sp. from calves were documented in tetracycline (100%, streptomycin (100% and compound sulphonamides (100% and enrofloxacin (50%. The high percentage (compound sulphonamides-100%, tetracycline-100% and streptomycin- 100% of multiresistant E. coli (isolates from dairy calves was noticed. There were no significant correlations between groups.
Full Text Available The vanC1 gene, which is chromosomally located, confers resistance to vancomycin and serves as a species marker for Enterococcus gallinarum. Enterococcus faecium TJ4031 was isolated from a blood culture and harbours the vanC1gene. Polymerase chain reaction (PCR assays were performed to detect vanXYc and vanTc genes. Only the vanXYc gene was found in the E. faecium TJ4031 isolate. The minimum inhibitory concentrations of vancomycin and teicoplanin were 2 µg/mL and 1 µg/mL, respectively. Real-time reverse transcription-PCR results revealed that the vanC1and vanXYc genes were not expressed. Pulsed-field gel electrophoresis and southern hybridisation results showed that the vanC1 gene was encoded in the chromosome. E. faecalis isolated from animals has been reported to harbour vanC1gene. However, this study is the first to report the presence of the vanC1gene in E. faecium of human origin. Additionally, our research showed the vanC1gene cannot serve as a species-specific gene of E. gallinarum and that it is able to be transferred between bacteria. Although the resistance marker is not expressed in the strain, our results showed that E. faecium could acquire the vanC1gene from different species.
Belguesmia, Y; Choiset, Y; Rabesona, H; Baudy-Floc'h, M; Le Blay, G; Haertlé, T; Chobert, J-M
The aim of this work was to study the antifungal properties of durancins isolated from Enterococcus durans A5-11 and of their chemically synthesized fragments. Enterococcus durans A5-11 is a lactic acid bacteria strain isolated from traditional Mongolian airag cheese. This strain inhibits the growth of several fungi including Fusarium culmorum, Penicillium roqueforti and Debaryomyces hansenii. It produces two bacteriocins: durancin A5-11a and durancin A5-11b, which have similar antimicrobial properties. The whole durancins A5-11a and A5-11b, as well as their N- and C-terminal fragments were synthesized, and their antifungal properties were studied. C-terminal fragments of both durancins showed stronger antifungal activities than other tested peptides. Treatment of D. hansenii LMSA2.11.003 strain with 2 mmol l(-1) of the synthetic peptides led to the loss of the membrane integrity and to several changes in the ultra-structure of the yeast cells. Chemically synthesized durancins and their synthetic fragments showed different antimicrobial properties from each other. N-terminal peptides show activities against both bacterial and fungal strains tested. C-terminal peptides have specific activities against tested fungal strain and do not show antibacterial activity. However, the C-terminal fragment enhances the activity of the N-terminal fragment in the whole bacteriocins against bacteria. © 2012 The Society for Applied Microbiology.
Shekh Raeesh M
Full Text Available Abstract Background Because Candida albicans is resistant to several antifungal antibiotics, there is a need to identify other less toxic natural products, particularly antimicrobial proteins, peptides or bacteriocin like inhibitory substances. An attempt has been made to purify and characterise an anti-Candida compound produced by Enterococcus faecalis. Results An anti-Candida protein (ACP produced by E. faecalis active against 8 C. albicans strains was characterised and partially purified. The ACP showed a broad-spectrum activity against multidrug resistant C. albicans MTCC 183, MTCC 7315, MTCC 3958, NCIM 3557, NCIM 3471 and DI. It was completely inactivated by treatment with proteinase K and partially by pronase E. The ACP retained biological stability after heat-treatment at 90°C for 20 min, maintained activity over a pH range 6–10, and remained active after treatment with α-amylase, lipase, organic solvents, and detergents. The antimicrobial activity of the E. faecalis strain was found exclusively in the extracellular filtrate produced in the late logarithmic growth phase. The highest activity (1600 AU mL-1 against C. albicans MTCC 183 was recorded at 48 h of incubation, and activity decreased thereafter. The peptide showed very low haemagglutination and haemolytic activities against human red blood cells. The antimicrobial substance was purified by salt-fractionation and chromatography. Partially purified ACP had a molecular weight of approximately 43 KDa in Tricine-PAGE analysis. The 12 amino acid N terminal sequence was obtained by Edman degradation. The peptide was de novo sequenced by ESI-MS, and the deduced combined sequence when compared to other bacteriocins and antimicrobial peptide had no significant sequence similarity. Conclusions The inhibitory activity of the test strain is due to the synthesis of an antimicrobial protein. To our knowledge, this is the first report on the isolation of a promising non-haemolytic anti
L. A. Chavez
Full Text Available El consumo de probióticos se ha asociado con mejoras en algunos parámetros productivos como la conversión alimenticia y la ganancia de peso vivo, lo que se ve reflejado en el desarrollo y salud de las aves. El objetivo de este trabajo fue evaluar la influencia de la inclusión de cepas probióticas en la alimentación de pollos de engorde sobre parámetros productivos de importancia económica. Se utilizaron 180 pollos machos (Cobb de un día de edad, alimentados con cinco dietas: dieta comercial con y sin la adición de antibióticos, y a esta última se le adicionó una de tres diferentes cepas probióticas (Lactobacillus casei, Lactobacillus acidophilus o Enterococcus faecium en el agua de bebida (108 UFC/ml durante 42 días. Se evaluaron parámetros zootécnicos: consumo de alimento, peso corporal, conversón alimenticia (CA y ganancia de peso (GDP; e indicadores productivos: supervivencia, factor de eficiencia americana (FEA, índice productivo (IP, eficiencia europea (EE y eficiencia alimenticia (EA. El diseño estadístico utilizado fue de bloques al azar. La inclusión de probióticos, específicamente E. faecium, mejoró parámetros productivos (P < 0,05 como peso (2.730 g, conversión (1,55, GDP (53,59 g/día, FEA (172, IP (393, EE (400 y EA (63,11%. Por todo lo anterior, la utilización de probióticos, especialmente E. faecium, puede ser considerada como factor promotor de crecimiento durante todo el ciclo de producción del ave debido a que demostró tener efectos positivos, tanto en el desempeño productivo, como en el rendimiento económico del lote.
Tennert, Christian; Feldmann, Katharina; Haamann, Edwina; Al-Ahmad, Ali; Follo, Marie; Wrbas, Karl-Thomas; Hellwig, Elmar; Altenburger, Markus J
To determine the antibacterial effect of photodynamic Therapy on Enterococcus faecalis (E. faecalis) biofilms in experimentally infected human root canals in primary infections and endodontic retreatments. One hundred and sixty single-rooted extracted teeth with one root canal were prepared using ProTaper instruments. Seventy specimens were left without root canal filling and autoclaved. The root canals of another 70 specimens were filled with Thermafil and AH Plus and the root canal fillings were removed after 24 hours using ProTaper D files and plasma sterilized. The specimens were infected with a clinical isolate of E. faecalis for 72 hours. Samples were taken using sterile paper points to determine the presence of E. faecalis in the root canals. The specimens were randomly divided into groups according to their treatment with 20 teeth each and a control. In the PDT group the teeth were treated using PDT, consisting of the photosensitizer toluidine blue and the PDT light source at 635 nm. In the NaOCl (sodium hypochlorite) group the root canals were rinsed with 10 mL of 3% NaOCl. In the NaOCl-PDT group the root canals were rinsed with 10 mL of 3% of sodium hypochlorite and then treated with PDT. Samples were taken after treatments using sterile paper points. Additionally, remaining root canal filling material was recovered from the root canal walls. Survival fractions of the samples were calculated by counting colony-forming units. A one-way analysis of variance (ANOVA) was applied to the data to assess the effect of different treatment techniques. Antimicrobial treatment of root canals caused a significant reduction of bacterial load in all groups. NaOCl irrigation eliminated E. faecalis most effectively. PDT alone was less effective compared to NaOCl irrigation and the combination of NaOCl irrigation and PDT. CFU levels recovered from the filling material after NaOCl irrigation of the root canals were 10fold higher compared to PDT and the combination of Na
Amachawadi, Raghavendra G; Giok, Felicia; Shi, Xiaorong; Soto, Jose; Narayanan, Sanjeev K; Tokach, Mike D; Apley, Mike D; Nagaraja, T G
Probiotics, an antibiotic alternative, are widely used as feed additives for performance benefits in cattle and swine production systems. Among bacterial species contained in probiotics, Enterococcus faecium is common. Antimicrobial resistance (AMR), particularly multidrug resistance, is a common trait among enterococci because of their propensity to acquire resistance and horizontally transfer AMR genes. Also, E. faecium is an opportunistic pathogen, and in the United States, it is the second most common nosocomial pathogen. There has been no published study on AMR and virulence potential in E. faecium contained in probiotic products used in cattle and swine in the United States. Therefore, our objectives were to determine phenotypic susceptibilities or resistance to antimicrobials, virulence genes (asa1, gelE, cylA, esp, and hyl) and assess genetic diversity of E. faecium isolated from commercial products. Twenty-two commercially available E. faecium-based probiotic products used in cattle (n = 13) and swine (n = 9) were procured and E. faecium was isolated and species confirmed. Antimicrobial susceptibility testing to determine minimum inhibitory concentrations was done by micro-broth dilution method using National Antimicrobial Resistance Monitoring Systems Gram-positive Sensititre panel plate (CMV3AGPF), and categorization of strains as susceptible or resistant was as per Clinical Laboratory and Standards Institute's guidelines. E. faecium strains from 7 products (3 for swine and 4 for cattle) were pan-susceptible to the 16 antimicrobials tested. Strains from 15 products (6 for swine and 9 for cattle) exhibited resistance to at least one antimicrobial and a high proportion of strains was resistant to lincomycin (10/22), followed by tetracycline (4/22), daptomycin (4/22), ciprofloxacin (4/22), kanamycin (3/22), and penicillin (2/22). Four strains were multidrug resistant, with resistant phenotypes ranging from 3 to 6 antimicrobials or class. None of the E
Perumal, Venkatesh; Repally, Ayyanna; Dasari, Ankaiah; Venkatesan, Arul
A novel bacteriocin produced by avian duck isolated lactic acid bacterium Enterococcus faecalis DU10 was isolated. This bacteriocin showed a broad spectrum of antibacterial activity against important food-borne pathogens and was purified by size exclusion chromatography followed by reverse-phase high-performance liquid chromatography in a C-18 column. Tricine-SDS PAGE revealed the presence of a band with an estimated molecular mass of 6.3 kDa. The zymogram clearly linked the antimicrobial activity with this band. This result was further confirmed by mass-assisted laser desorption ionization time-of-flight mass spectrometry, since a sharp peak corresponding to 6.313 kDa was detected and the functional groups were revealed by Fourier transform infrared spectroscopy. Bacteriocin DU10 activity was found sensitive to proteinase-K and pepsin and partially affected by trypsin and α-chymotrypsin. The activity of bacteriocin DU10 was partially resistant to heat treatments ranging from 30 to 90°C for 30 min. It also withstood a treatment at 121°C for 10 min. Cytotoxicity of bacteriocin DU10 by methyl-thiazolyl-diphenyl-tetrazolium bromide assay showed that the viability of HT-29 and HeLa cells decreased 60 ± 0.7% and 43 ± 4.8%, respectively, in the presence of 3,200 AU/mL of bacteriocin. The strain withstood 0.3% w/v of bile oxgall and pH 2 affected the bacterial growth between 2 and 4 hr of incubation. Adhesion properties examined with HT-29 cell line showed 69.85% initial population of strain E. faecalis DU10, which was found to be strongly adhered to this cell line. These results conclude bacteriocin DU10 may be used as a potential biopreservative and E. faecalis DU10 may be used as a potential probiont to control Salmonella infections.
Kobayashi, Nobumichi; Nagashima, Shigeo
We carried out the first study of Enterococcus faecalis clinical isolates in Cuba by multilocus sequence typing linking the molecular typing data with the presence of virulence determinants and the antibiotic resistance genes. A total of 23 E. faecalis isolates recovered from several clinic sources and geographic areas of Cuba during a period between 2000 and 2005 were typed by multilocus sequence typing. Thirteen sequence types (STs) including five novel STs were identified, and the ST 64 (clonal complex [CC] 8), ST 6 (CC2), ST 21(CC21), and ST 16 (CC58) were found in more than one strain. Sixty-seven percent of STs corresponded to STs reported previously in Spain, Poland, and The Netherlands, and other STs (ST115, ST64, ST6, and ST40) were genetically close to those detected in the United States. Prevalence of both antimicrobial resistance genes [aac(6′)-aph(2″), aph(3′), ant(6), ant(3″)(9), aph(2″)-Id, aph(2″)-Ic, erm(B), erm(A), erm(C), mef(A), tet(M), and tet(L)] and virulence genes (agg, gelE, cylA, esp, ccf, and efaAfs) were examined by polymerase chain reaction. Aminoglycoside resistance genes aac(6′)-Ie-aph(2″)-Ia, aph(3′), ant(6), ant(3″)(9) were more frequently detected in ST6, ST16, ST23, ST64, and ST115. The multidrug resistance was distributed to all STs detected, except for ST117 and singleton ST225. The presence of cyl gene was specifically linked to the ST64 and ST16. Presence of the esp, gel, and agg genes was not specific to any particular ST. This research provided the first insight into the population structure of E. faecalis in Cuba, that is, most Cuban strains were related to European strains, whereas others to U.S. strains. The CC2, CC21, and CC8, three of the biggest CCs in the world, were evidently circulating in Cuba, associated with multidrug resistance and virulence traits. PMID:19857135
Gomes Luiz Humberto
Full Text Available A simple DNA isolation method was developed with routine chemicals that yields high quality and integrity preparations when compared to some of the most well known protocols. The method described does not require the use of lysing enzymes, water bath and the DNA was obtained within 40 minutes The amount of nucleic acid extracted (measured in terms of absorbancy at 260 nm from strains of Xanthomonas spp., Pseudomonas spp. and Erwinia spp. was two to five times higher than that of the most commonly used method.
A. G. S. Cuthbertson
Full Text Available Very little information exists in regards to the control options available for potato flea beetles, Epitrix spp. This short review covers both chemical and ecological options currently available for control of Epitrix spp. Synthetic pyrethroids are the weapon of choice for the beetles. However, the impetus in integrated pest management is to do timely (early-season applications with something harsh which will give long-term protection at a time when there are not a lot of beneficials in the field. Finding the balance for control of Epitrix spp. is proving difficult.
Jonatas Campos Almeida
Full Text Available The purpose of this study was to investigate the occurrence of Cryptosporidium spp. and Giardia spp. in a public water-treatment system. Samples of raw and treated water were collected and concentrated using the membrane filtration technique. Direct Immunofluorescence Test was performed on the samples. DNA extraction using a commercial kit was performed and the DNA extracted was submitted to a nested-PCR reaction (n-PCR and sequencing. In the immunofluorescence, 2/24 (8.33% samples of raw water were positive for Giardia spp.. In n-PCR and sequencing, 2/24 (8.33% samples of raw water were positive for Giardia spp., and 2/24 (8.33% samples were positive for Cryptosporidium spp.. The sequencing showed Cryptosporidium parvum and Giardia duodenalis DNA. In raw water, there was moderate correlation among turbidity, color and Cryptosporidium spp. and between turbidity and Giardia spp.. The presence of these protozoans in the water indicates the need for monitoring for water-treatment companies.
Ruiz-Garbajosa, P.; de Regt, M.; Bonten, M.; Baquero, F.; Coque, T. M.; Canton, R.; Harmsen, H. J. M.; Willems, Rob J. L.
Enterococcus faecium belonging to the polyclonal subcluster CC17, with a typical ampicillin-resistant E. faecium (AREfm) phenotype, have become prevalent among nosocomial infections around the world. High-density intestinal AREfm colonization could be one of the factors contributing to the
The U.S. Environmental Protection Agency (EPA) has provided recommended beach advisory values in its 2012 recreational water quality criteria (RWQC) for states wishing to use quantitative polymerase chain reaction (qPCR) for the monitoring of Enterococcus fecal indicator bacteria...
Prichula, Janira; Campos, Fabricio Souza; Pereira, Rebeca Inhoque; Cardoso, Leonardo Almansa; Wachholz, Guilherme Raffo; Pieta, Luiza; Mariot, Roberta Fogliatto; de Moura, Tiane Martin; Tavares, Maurício; d’Azevedo, Pedro Alves; Frazzon, Ana Paula Guedes
Enterococcus faecalis strains have a ubiquitous nature that allows them to survive in different niches. Studies involving enterococci isolated from marine animals are scarce. Therefore, in this study, we report the complete genome sequence of E. faecalis strain P8-1 isolated from feces of a Magellanic penguin on the south coast of Brazil. PMID:26769928
van der Waal, S. V.; van der Sluis, L. W. M.; Ozok, A. R.; Exterkate, R. A. M.; van Marle, J.; Wesselink, P. R.; de Soet, J. J.
van der Waal SV, van der Sluis LWM, Ozok AR, Exterkate RAM, van Marle J, Wesselink PR, de Soet JJ. The effects of hyperosmosis or high pH on a dual-species biofilm of Enterococcus faecalis and Pseudomonas aeruginosa: an in vitro study. International Endodontic Journal, 44, 11101117, 2011. Aim To
Paganelli, Fernanda L.; Huebner, Johannes; Singh, Kavindra V.; Zhang, Xinglin; Van Schaik, Willem; Wobser, Dominique; Braat, Johanna C.; Murray, Barbara E.; Bonten, Marc J M; Willems, Rob J L; Leavis, Helen L.
Enterococcus faecium is a common cause of nosocomial infections, of which infective endocarditis is associated with substantial mortality. In this study, we used a microarray-based transposon mapping (M-TraM) approach to evaluate a rat endocarditis model and identified a gene, originally annotated
Licht, Tine Rask; Laugesen, D.; Jensen, Lars Bogø
A new animal model, the streptomycin-treated mini-pig, was developed in order to allow colonization of defined strains of Enterococcus faecalis in numbers sufficient to study plasmid transfer. Transfer of the pheromone-inducible pCF10 plasmid between streptomycin-resistant strains of E. faecalis OG...
Gutierrez, Jorge; Larsen, Rasmus; Cintas, Luis M.; Kok, Jan; Hernandez, Pablo E.
Enterocin P (EntP), a sec-dependent bacteriocin from Enterococcus faecium P13, was produced by Lactococcus lactis. The EntP structural gene (entP) with or without the EntP immunity gene (entiP) was cloned in (1), plasmid pMG36c under control of the lactococcal constitutive promoter P-32, (2) in
Waar, Karola; van der Mei, Henderina; Harmsen, Hermie JM; de Vries, Jacob; Atema-Smit, Jelly; Degener, John E; Busscher, Hendrik
Enterococcus faecalis is one of the leading causes of hospital-acquired infections, and indwelling medical devices are especially prone to infection. E faecalis expressing aggregation substance (Agg) adheres to biomaterial surfaces by means of positive cooperativity, i.e. the ability of one adhering
Hammerum, Anette Marie; Fussing, Vivian; Aarestrup, Frank Møller
Forty-eight vancomycin-resistant and 35 vancomycin-sensitive Danish Enterococcus faecium isolates obtained from pigs, chickens and humans, as well as the human vanA reference isolate BM4147, were characterized by EcoRI RiboPrinting and Smal pulsed-field gel electrophoresis. RiboPrinting of the 84...
de Diego-Diaz, Beatriz; Treu, Laura; Campanaro, Stefano
The whole genome sequence of Enterococcus mundtii strain EM01 is reported here. The isolate proved to be the cause of flacherie in Bombyx mori To date, the genomes of 11 other E. mundtii strains have been sequenced. EM01 is the only strain that displayed active pathological effects on its...
Guzman Prieto, Ana M.; Urbanus, Rolf T.; Zhang, Xinglin; Bierschenk, Damien; Koekman, C. Arnold; van Luit-Asbroek, Miranda; Ouwerkerk, Janneke P.; Pape, Marieke; Paganelli, Fernanda L.; Wobser, Dominique; Huebner, Johannes; Hendrickx, Antoni P. A.; Bonten, Marc J. M.; Willems, Rob J. L.; van Schaik, Willem
Enterococcus faecium is a commensal of the mammalian gastrointestinal tract, but is also found in non-enteric environments where it can grow between 10 degrees C and 45 degrees C. E. faecium has recently emerged as a multi-drug resistant nosocomial pathogen. We hypothesized that genes involved in
Stojanović Nikola; Krunić Jelena; Popović Branka; Stojičić Sonja; Živković Slavoljub
Introduction. Because apical periodontitis is recognizably an infectious disease, elimination or reduction of intracanal bacteria is of utmost importance for optimum treatment outcome. Objective. The prevalence of Enterococcus faecalis and Porphyromonas gingivalis in infected root canals was studied Also, the effect of endodontic therapy by using intracanal medicaments, calcium hydroxide paste (CH) or gutta-percha points containing calcium hydroxide (CH-GP)...
Peters, Remco P. H.; van Agtmael, Michiel A.; Gierveld, Sonja; Danner, Sven A.; Groeneveld, A. B. Johan; Vandenbroucke-Grauls, Christina M. J. E.; Savelkoul, Paul H. M.
Direct detection of bacterial DNA in blood offers a fast alternative to blood culture and is presumably unaffected by the prior use of antibiotics. We evaluated the performance of two real-time PCR assays for the quantitative detection of Staphylococcus aureus bacteremia and for Enterococcus
Application of the biosurfactants produced by Bacillus spp. (SH 20 and SH 26) and P. aeruginosa SH 29 isolated from the rhizosphere soil of an Egyptian salt marsh plant for the cleaning of oil - contaminataed vessels and enhancing the biodegradat.
Jun 2, 2014 ... Introduction. Application of antibiotics in the treatment of bacterial ... Keywords: Bacillus spp, antibacterial activity, eyes pathogens, conjunctiva. African Health ... ml of respective test organism and allowed to dry. In the agar ...
Sofya K. Garushyants
Full Text Available While most endosymbiotic bacteria are transmitted only vertically, Holospora spp., an alphaproteobacterium from the Rickettsiales order, can desert its host and invade a new one. All bacteria from the genus Holospora are intranuclear symbionts of ciliates Paramecium spp. with strict species and nuclear specificity. Comparative metabolic reconstruction based on the newly sequenced genome of Holospora curviuscula, a macronuclear symbiont of Paramecium bursaria, and known genomes of other Holospora species shows that even though all Holospora spp. can persist outside the host, they cannot synthesize most of the essential small molecules, such as amino acids, and lack some central energy metabolic pathways, including glycolysis and the citric acid cycle. As the main energy source, Holospora spp. likely rely on nucleotides pirated from the host. Holospora-specific genes absent from other Rickettsiales are possibly involved in the lifestyle switch from the infectious to the reproductive form and in cell invasion.
The mucosal barriers of catfish (Ictalurus spp.) constitute the first line of defense against pathogen invasion while simultaneously carrying out a diverse array of other critical physiological processes, including nutrient adsorption, osmoregulation, waste excretion, and environmental sensing. Catf...
... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3870 Trypanosoma... consist of antigens and antisera used in serological tests to identify antibodies to Trypanosoma spp. in...
... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3065 Bordetella... serological tests to identify Bordetella spp. from cultured isolates or directly from clinical specimens. The...
Gore, P.S.; Raveendran, O.; Unnithan, R.V.
Occurrence, isolation and oxidative activity of Thiobacilli spp. from some sandy beaches of Kerala are reported. These organisms were encountered in polluted beaches and were dominant during monsoon in all the beaches...
Sagor, Rakibul Hasan; Alsunaidi, Mohammad A.; Ooi, Boon S.
The non-linear propagation of Surface-Plasmon-Polaritons (SPP) in single interface of metal and chalcogenide glass (ChG) is considered. A time domain simulation algorithm is developed using the Finite Difference Time Domain (FDTD) method
Havelaar AH; During M; Versteegh JFM
Door middel van onderzoek van zes kunstmatig besmette gesimuleerde monsters drinkwater ( 4 Aeromonas spp., 1 Klebsiella oxytoca, 1 Pseudomonas aeruginosa) werd een vergelijking gemaakt van de resultaten van Aeromonas onderzoek in 14 laboratoria. De tellingen in de deelnemende laboratoria
had significant (P≤ 0.05) effect on bean plant girth, number of leaves, number of branches, mean number of flowers, total fresh ... Beans (Phaseolus spp) belong to one of several genera .... Meng (2016), that found that applying coffee pulp.
Dec 15, 2009 ... Available online at http://www.academicjournals.org/AJB. ISSN 1684–5315 ... Molecular characterization of Azotobacter spp. nifH .... MATERIALS AND METHODS ..... rapidly expanding and is currently composed of over.
Full Text Available Cronobacter spp. (Enterobacter sakazakii is an opportunistic bacterial pathogen capable of causing disease and even fatalities in newborn infants within the first weeks of life if consumed as part of the diet. Premature and immunocompromised newborn infants are at particular risk. The microorganism has been isolated from a variety of foods including contaminated infant milk formula powder and milk powder substitute. The study aimed to evaluate the level of microbiological contamination in 47 samples of mozzarella cheese made with cow’s milk collected from artisan cheese producers in Southern Italy. Samples were collected from commercial sales points and underwent qualitative and quantitative microbiological analyses to test for the bacterial contaminants most commonly found in milk and cheese products. The 47 samples underwent qualitative and quantitative microbiological tests according to ISO UNI EN standards. Analyses focused on Staphylococcus aures, Salmonella spp., Listeria monocytogenes, Pseudomonas spp., E. coli, Yersinia spp., total coliforms and Cronobacter sakazakii. The ISO/TS 22964:2006 method was used to investigate possible contamination by C. sakazakii. Biochemical identification was carried out using an automated system for identification and susceptibility tests. None of the samples examined resulted positive for Salmonella spp. or Listeria spp. Only one sample resulted positive for Staphylococcus aureus. Pseudomonas spp. was isolated in 10 (21% of 47 samples. High levels of total coliforms were found in 10 of 47 samples. Cronobacter spp. (Enterobacter sakazakii was isolated in one sample. This is the first study to confirm isolation of C. sakazakii in artisan mozzarella cheese made from cow’s milk. The presence of C. sakazakii could be related to external contamination during the phases of production or to the use of contaminated milk. Since mozzarella is recommended in the diet of children and adults of all ages, this
The research about potential of some probiotic isolates as an antibacterial on the growth of Vibrio spp had been done. This research aimed to know the antibacterial potency from some isolates probiotic on the growth of Vibrio spp. This research to tested the inhibition on the three species of Vibrio that are Vibrio harveyi, Vibrio prahaemolyticus, and Vibrio cholerae using agar diffusion method. Probiotic isolates come from lactic acid bacteria group that provide beneficial effects on health ...
Full Text Available Objective. To characterize and identify yeasts of the genus Malassezia by phenotypic features. Materials and methods. First, the macroscopic and microscopic morphological characteristics were described. In addition we performed biochemical and physiological assays as Tweens and Cremophor, including more. Results. Our results evidenced of 105 isolates obtained from dogs diagnosed with external otitis, it was possible to identify two distinct species from 46 isolates within the Malassezia genus: 36.19% (n=38 were identified as M. pachydermatis and 7.62% (n=8 as M. furfur. According to phenotypic patterns the remaining 56.19% (n=59 were reported as Malassezia spp., possibly corresponding to M. furfur and/or M. pachydermatis. Conclusions. Results emphasize the necessity to characterize according to species. It is not feasible to define Malassezia by species based on morphological, biochemical, and physiological findings. Therefore, molecular genotyping should be performed to identify markers allowing a more precise isolate identification. This would broaden our epidemiological knowledge regarding different species involved in canine otitis pathologies.
Full Text Available Prosopis is a commercially important plant genus, which has been used since ancient times, particularly for medicinal purposes. Traditionally, Paste, gum, and smoke from leaves and pods are applied for anticancer, antidiabetic, anti-inflammatory, and antimicrobial purposes. Components of Prosopis such as flavonoids, tannins, alkaloids, quinones, or phenolic compounds demonstrate potentials in various biofunctions, such as analgesic, anthelmintic, antibiotic, antiemetic, microbial antioxidant, antimalarial, antiprotozoal, antipustule, and antiulcer activities; enhancement of H+, K+, ATPases; oral disinfection; and probiotic and nutritional effects; as well as in other biopharmaceutical applications, such as binding abilities for tablet production. The compound juliflorine provides a cure in Alzheimer disease by inhibiting acetylcholine esterase at cholinergic brain synapses. Some indirect medicinal applications of Prosopis spp. are indicated, including antimosquito larvicidal activity, chemical synthesis by associated fungal or bacterial symbionts, cyanobacterial degradation products, “mesquite” honey and pollens with high antioxidant activity, etc. This review will reveal the origins, distribution, folk uses, chemical components, biological functions, and applications of different representatives of Prosopis.
Full Text Available Brachiaria species are the most important grasses for cattle production in Brazil. However, a limiting factor for the use of Brachiaria spp. is their toxicity. Most outbreaks of hepatogenous photosensitization are caused by B. decumbens; however B. brizantha, B. humidicola and B. ruziziensis can also cause poisoning. The poisoning affects cattle, sheep, goats and buffalo. Sheep are more susceptible than other animal species and the young are more susceptible than adults. There are differences in susceptibility among animals of the same species and it has been suggested that this resistance is genetic. Also has been suggested that buffalo and probably some sheep are resilient, i.e. when poisoned these animals have histologic lesions and high GGT serum concentrations, but do not show clinical signs. In general, saponin concentrations are higher in growing plants, but outbreaks occur all over the year, probably due to unexplained rise in saponin concentration in the plant. A clinical syndrome of progressive weight loss and death, without photosensitization, has been reported in cattle poisoned by B. decumbens. Main preventive measures are based on the selection of resistant or resilient animals and on the development of Brachiaria species or varieties with low saponin concentration.
Longyan, G.; Xinguo, L.; Lingxia, W.; Xuefei, J.
Being one of the important tropical and subtropical fruit trees, banana (Musa spp.) belongs to the family Musaceae and the order Scitaminae with two genera, Musa and Ensete. In a field survey, research team has discovered a potential banana mutant strain HD-1 with a sound economic value. The results of the finding are as follows: based on Simmonds classification, the pseudostem of banana strain HD-1 is relatively short and purplish red; its upright outward petiole groove has red edges and wraps its pseudostem loosely. Its ploidy is 3, AAA type. Karyotype analysis shows that the number of chromosomes is 33, the karyotype formula is 2n=3x=33=2L + 3 M2 + 4 M1 + 2 S, HD-1 is classified as 1B type. With the help of ISSR molecular markers, we find thatbanana HD-1 has the closest relationship with Pubei and Tianbao dwarf banana; the similarity coefficient is 0.81. In an artificial simulation tests of cold, drought and salt resistance environment changes of physiological and biochemical indexes indicate that HD-1 exhibits stronger defense capability than Brazil banana. By way of inoculation with injury of root dipping method, we respectively treat two kinds of banana seedlings inoculated Banana Fusarium wilt race 4 small species. The results show that their resistance evaluation scores are 3 and 4, disease levels are susceptible and high sensitivity respectively. We conclude that HD-1 has stronger resistance ability to Fusarium wilt than Brazil banana. (author)
Greenhouse and field experiments were undertaken to determine the possibility of using soil amendments with different C:N levels or applied at different rates and times in the control of root-knot nematodes (Meloidogyne spp.)in tomato c.v Cal J.A naturally infested field was used while artificial inoculation was done in the greenhouse. Root galling was rated on a scale of 0-10, nematode population was estimated by counting second stage juveniles extracted from 200 cm 3 soil and fruit yields were recorded at the end of the season. Nematode population densities and galling indices were significantly (P< or=0.05) lower in amended soils compared to the control. Application of the amendments also resulted in significant (P< or=0.05) increase in yields. Chicken manure, compost manure, neem products and pig manure were were the most effective amendments. Fresh chicken manure had a more suppressive effect on nematode than when the manure was decomposed within or outside a nematode infested field. A general decrease in juvenile populations and galling was observed with increase of organic amendments applied
Bento, C S; de Souza, A G; Sudré, C P; Pimenta, S; Rodrigues, R
This study aimed to identify Capsicum genotypes with resistance to bacterial spot (BS), anthracnose and Pepper yellow mosaic virus (PepYMV). Fifty-four genotypes of Capsicum spp were evaluated. Resistance reaction against BS was evaluated using three replicates, testing hypersensitivity and quantitative resistance in leaves. After evaluation, inoculated leaves were detached from the plants, being then cultivated until reproductive stage for evaluations anthracnose resistance in immature and mature fruit, totalizing 18 fruits per genotype. For PepYMV resistance was performed with five replications. Each genotype reaction was evaluated by a scoring scale, using the area under the disease progress curve for each pathosystem, and incubation period for the three systems. The latent period was evaluated only for the pathosystem Capsicum-Colletotrichum gloeosporioides. Means were grouped by the Scott-Knott test. Measures of dissimilarity matrix among the genotypes were obtained by Gower's algorithm and the grouping was obtained by the UPGMA clustering method. The accessions belonging to the Capsicum frutescens were the most susceptible to the three diseases. At least one genotype of Capsicum baccatum var. pendulum, Capsicum annuum, and Capsicum chinense showed resistance potential to BS and PepYMV, for use in breeding programs. The accession UENF 1381 (C. annuum) was resistant to the three pathogens.
Gomes, Carmen; Moreira, Rosana G; Castell-Perez, Elena
The FDA recently approved irradiation treatment of leafy greens such as spinach up to 1 kGy; however, it is important to reduce the dose required to decontaminate the produce while maintaining its quality. Thus, the objectives of this study were: (1) to assess the radiation sensitivities of Salmonella spp. and Listeria spp. inoculated in ready-to-eat baby spinach leaves under modified atmosphere packaging (MAP) and irradiated using a 1.35-MeV Van de Graff accelerator (the leaves were irradiated both at room temperature and at -5 °C); and (2) to understand and optimize the synergistic effect of MAP and irradiation by studying the radiolysis of ozone formation under different temperatures, the effect of dose rate on its formation, and its decomposition. Results showed that increased concentrations of oxygen in the packaging significantly increased the radiation sensitivity of the test organisms, ranging from 7% up to 25% reduction in D(10)-values. In particular, radiosensitization could be effected (P radiation under modified atmosphere packaging (100% O(2) and N(2):O(2)[1:1]) may be a viable tool for reducing microbial populations or eliminating Salmonella spp. and Listeria spp. from baby spinach. A suggested treatment to achieve a 5-log reduction of the test organisms would be irradiation at room temperature under 100% O(2) atmosphere at a dose level of 0.7 kGy. Practical Application: Decontamination of minimally processed fruits and vegetables from food-borne pathogens presents technical and economical challenges to the produce industry. Internalized microorganisms cannot be eliminated by the current procedure (water-washed or treated with 200-ppm chlorine). The only technology available commercially is ionizing radiation; however, the actual radiation dose required to inactivate pathogens is too high to be tolerated by the product without unwanted changes. This study shows a new approach in using MAP with 100% O(2), which is converted to ozone to radiosensitize
Gelpud Chaves Cristian
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El cultivo del lulo (Solanum quitoense L. presenta una disminución en su productividad, debido al ataque de patógenos como el nematodo del nudo radical Meloidogyne spp., en el Departamento de Nariño (Colombia, se han reportado incidencias cercanas al 79%, y pérdidas del 50%. En la presente investigación, se colectaron 45 genotipos de (Solanum quitoense L. en los Departamentos de Nariño y Putumayo y 4 genotipos silvestres (S. mammosum, S. hirtum, S. marginatum y S. umbellatum buscando fuentes de resistencia al nematodo. Se inocularon 9 plantas de cada genotipo de dos meses de edad con 10000 huevos de Meloidogyne spp., dejando tres testigos por cada material. Las variables evaluadas fueron: altura de planta, severidad, incidencia, peso fresco (tallo y raíz y especies prevalentes de Meloidogyne spp. Se hizo una clasificación de genotipos mediante escala de resistencia y regresión entre la severidad y las demás variables para establecer el efecto de Meloidogyne spp. sobre los genotipos de planta. Los resultados mostraron 100% de incidencia del nematodo en todos los genotipos, 2.04% genotipos resistentes, 34.7% moderadamente resistentes, 42.8% moderadamente susceptibles, 18.3% susceptibles, y 2.04% altamente susceptibles. El genotipo SQbr05 resistente, no se vio afectado por la severidad, al contrario SQbc04 genotipo susceptible, mostró reducciones significativas en peso fresco de tallo y raIz, (R2 = 0.71 y 0.98,el genotipo silvestre (S. mammosum es altamente susceptible, Meloidogyne incognita presentó 55.31% de presencia. El genotipo SQbr05 es promisorio para ser evaluado en campo.
Samantha Yuri Oshiro Branco Valadas
Full Text Available South American opossums are the definitive hosts of Sarcocystis neurona, Sarcocystis falcatula, Sarcocystis speeri and Sarcocystis lindsayi. The sporocysts of these species of Sarcocystis are morphologically similar and methods like infectivity and pathogenicity for intermediate hosts (immunodeficient mice and psittacine birds and molecular tools are used for identification. Opossums are synanthropic wild animals, and widely distributed in Brazilian territory. Previous studies have shown high environmental contamination with S. neurona sporocysts in several Brazilian regions. This paper reviews information on Sarcocystis spp. shed by various opossum species and its occurrence in Brazil.
Alves, Carlos Roberto; Souza, Raquel Santos de; Charret, Karen Dos Santos; Côrtes, Luzia Monteiro de Castro; Sá-Silva, Matheus Pereira de; Barral-Veloso, Laura; Oliveira, Luiz Filipe Gonçalves; da Silva, Franklin Souza
Serine proteases have significant functions over a broad range of relevant biological processes to the Leishmania spp lifecycle. Data gathered here present an update on the Leishmania spp serine proteases and the status of these enzymes as part of the parasite degradome. The serine protease genes (n = 26 to 28) in Leishmania spp, which encode proteins with a wide range of molecular masses (35 kDa-115 kDa), are described along with their degrees of chromosomal and allelic synteny. Amid 17 putative Leishmania spp serine proteases, only ∼18% were experimentally demonstrated, as: signal peptidases that remove the signal peptide from secretory pre-proteins, maturases of other proteins and with metacaspase-like activity. These enzymes include those of clans SB, SC and SF. Classical inhibitors of serine proteases are used as tools for the characterization and investigation of Leishmania spp. Endogenous serine protease inhibitors, which are ecotin-like, can act modulating host actions. However, crude or synthetic based-natural serine protease inhibitors, such as potato tuber extract, Stichodactyla helianthus protease inhibitor I, fukugetin and epoxy-α-lapachone act on parasitic serine proteases and are promising leishmanicidal agents. The functional interrelationship between serine proteases and other Leishmania spp proteins demonstrate essential functions of these enzymes in parasite physiology and therefore their value as targets for leishmaniasis treatment. Copyright © 2017 Elsevier Inc. All rights reserved.
Razina, I N; Chesnokova, M G; Nedoseko, V B
The aim of the study was to assess the correlation of Candida spp. incidence in periodontal tissues with various clinical manifestations of chronic periodontal disease (CPD). Ninety patients with CPD were included in the study in which Candida spp. was evaluated in periodontal pockets content and gingival biopsy material. In severe CPD more Candida spp. were seen in gingival biopsy than in periodontal pockets (p=0.0006). Candida spp. incidence and quantity correlated directly with the disease grade showing incidence increase from 40 to 73.3% and quantity increase from 0.8±0.18 до 3.6±0.49 lg CFU/ml in light and severe CPD, correspondingly Candida spp. had statistically significant association with cyanotic gingival color (p=0.0018), tongue plaque and swelling (р=0.0042), lip exfoliation (р=0.0030), periodontal pockets depth >5 mm (р=0.0030), oral mucosa hyperemia (р=0.0157), alveolar bone destruction >1/2 of root length (р=0.0157). These data prove the relevance of Candida spp. and mycological assessment of gingival biopsy in CPD patients.
Full Text Available Soil microbes associated with rhizosphere are important for promoting plant growth and inducing resistance to diseases. The research was conducted to study the ability of Trichoderma spp. and Penicillium spp. isolated from rhizosphere in lowland swampy area for controlling damping-off disease caused by Rhizoctonia solani Khun. Trichoderma spp. and Penicillium spp. were cultured in bran, corn meal, and rice straw containing media and applied as inoculum to 2-weeks old seedlings. Application of two fungi isolates effectively induced resistance of chili plants to damping-off disease. Trichoderma spp. and Penicillium spp. were significantly reduced disease incidence by 61.5–100% to 46.2–100%, respectively and disease severity by 50–100% and 30–95.9%, respectively. This experiment showed the potential of Trichoderma spp. and Penicillium spp. as biocontrol agents to control damping-off disease on chili.
U.S. Environmental Protection Agency — Quantity of Legionella spp., Mycobacterium spp., Acanthamoeba,Vermamoeba vermiformis and Pseudomonas aeruginosa were estimated using qPCR methods. This dataset is...
Abdelfattah M. Selim
Full Text Available Abortion among dairy cattle is one of the major causes of economic losses in the livestock industry. This study describes a 1-step multiplex real-time polymerase chain reaction (PCR to detect Brucella spp., Leptospira spp. and Campylobacter foetus, these are significant bacteria commonly implicated in bovine abortion. ß-actin was added to the same PCR reaction as an internal control to detect any extraction failure or PCR inhibition. The detection limit of multiplex real-time PCR using purified DNA from cultured organisms was set to 5 fg for Leptospira spp. and C. foetus and to 50 fg for Brucella spp. The multiplex real-time PCR did not produce any non-specific amplification when tested with different strains of the 3 pathogens. This multiplex real-time PCR provides a valuable tool for diagnosis, simultaneous and rapid detection for the 3 pathogens causing abortion in bovine.
Viken, Knut Ole
Abstract The objectives of this study were (1) to develop models for estimation of stand-level tree biomass for spruce (picea spp.)- pine (pinus spp.)- and broadleaved-dominated forest in Norway and, (2) develop biomass expansion factors (BEFs; ratio of stem volume to biomass) which convert stem volume to whole tree biomass for Norwegian forest conditions. A dataset from a 5 year period (2006 – 2010) from the Norwegian National Forest Inventory (NFI) were used to develop the...
Cui, Yanyan; Zhang, Yan; Jian, Fuchun; Zhang, Longxian; Wang, Rongjun; Cao, Shuxuan; Wang, Xiaoxing; Yan, Yaqun; Ning, Changshen
Theileria spp. and Anaplasma spp., which are important tick-borne pathogens (TBPs), impact the health of humans and animals in tropical and subtropical areas. Theileria and Anaplasma co-infections are common in sheep and goats. Following alignment of the relevant DNA sequences, two primer sets were designed to specifically target the Theileria spp. 18S rRNA and Anaplasma spp. 16S rRNA gene sequences. Genomic DNA from the two genera was serially diluted tenfold for testing the sensitivities of detection of the primer sets. The specificities of the primer sets were confirmed when DNA from Anaplasma and Theileria (positive controls), other related hematoparasites (negative controls) and ddH 2 O were used as templates. Fifty field samples were also used to evaluate the utility of single PCR and duplex PCR assays, and the detection results were compared with those of the PCR methods previously published. An optimized duplex PCR assay was established from the two primer sets based on the relevant genes from the two TBPs, and this assay generated products of 298-bp (Theileria spp.) and 139-bp (Anaplasma spp.). The detection limit of the assay was 29.4 × 10 -3 ng per μl, and there was no cross-reaction with the DNA from other hematoparasites. The results showed that the newly developed duplex PCR assay had an efficiency of detection (P > 0.05) similar to other published PCR methods. In this study, a duplex PCR assay was developed that can simultaneously identify Theileria spp. and Anaplasma spp. in sheep and goats. This duplex PCR is a potentially valuable assay for epidemiological studies of TBPs in that it can detect cases of mixed infections of the pathogens. Copyright © 2017 Elsevier Inc. All rights reserved.
Leite, Diniz Pereira; Yamamoto, Ana Caroline Akeme; Amadio, Janaína Vasconcellos Ribeiro de Souza; Martins, Evelin Rodrigues; do Santos, Fábio Alexandre Leal; Simões, Sara de Almeida Alves; Hahn, Rosane Christine
Atmospheric air is the most common vehicle for the dispersion of fungi. Fungi belonging to the genera Aspergillus and Penicillium are cosmopolitan and are classified in the family Trichocomaceae. Species of the genera are commonly found in soil, decaying organic materials, animal feed, stored grains, and other materials. This study aimed to determine the taxonomic diversity of airborne fungi of the genera Aspergillus and Penicillium residing in the dust of library environments to contribute to current knowledge of these characteristic genera. Three libraries in the city of Cuiaba, State of Mato Grosso, Brazil, were selected as the study areas. A total of 168 samples were collected at randomized sites within each library in areas containing journals, archives, in study rooms, and in collection storage areas in two different periods, the dry season (n = 42) and the rainy season (n = 42). Samples were collected by exposing Petri dishes containing Sabouraud agar with chloramphenicol to the environmental air. Additional samples were collected with sterile swabs which were rubbed over the surface of randomly chosen books on the shelves; the swabs were subsequently incubated in the laboratory. The genus Aspergillus was highlighted as one of the principal airborne fungi present in indoor environments. Aspergillus spp was identified in 1,277 (89.6%) samples and Penicillium spp in 148 (10.4%). The dry period exhibited a greater number of isolates of the two taxons.
Full Text Available Tuberculosis and brucellosis remain important causes of morbidity and mortality in many countries, for the detection of both diseases requires efficient and sensitive tool for effectuate the diagnosis. This study was aimed to evaluate and compare the duplex PCR versus the nested PCR, for detection of Brucella spp. (BR and Mycobacterium spp. (TB. A total of 100 samples of tissues from tracheo-bronchial lymph nodes, bovine lung and bacterial isolate as positive controls were used. Were evaluated ten combinations of primers which were designed to flank the segment of the 16S rRNA sequence (RB and antigen gen MPB70 (TB, the best result for the Duplex PCR was obtained with the primers Bru-2F/Bru-2R for BR and Tub-1F/Tub-N-R for TB. The amplification of the products was 225 and 230-bp respectively. In order to compare the results of the proposed technique, all samples were initially analyzed and compared between PCR and nested PCR (Kappa, k = 0.85 and the concordance between Duplex PCR and nested PCR (k = 0.88 for the two bacteria was very good.
Fancelli, Marilene; Sanches, Nilton F.; Dantas, Jorge L.L.; Caldas, Ranulfo C.; Morales, Cinara F.G.
The papaya borer weevil, Pseudopiazurus papayanus (Marshall), is generally considered a secondary pest, but it has been reported in high infestations in Northeast Brazil. This work aimed at evaluating the occurrence of P. papayanus and reporting its infestation level in papaya genotypes kept at the germplasm bank of EMBRAPA Cassava and Tropical Fruits (Cruz das Almas, Bahia, Brazil). The number of larvae, pupae and adults found in each plant of 65 Carica spp. genotypes and of three Vasconcella spp. genotypes was registered in three to five plants of each genotype, by cutting the exsudating trunks lengthwise. Papaya borer weevil was found in C. papaya and V. cauliflora but not in those of V. quercifolia. Among the evaluated genotypes, 52.4% of those belonging to the Solo group were infested, against 25.0% of the Formosa group. Larval infestation was the best criterion for sorting out genotypes concerning this insect infestation. This is also the first occurrence of the papaya borer weevil . (author)
Full Text Available Abstract Pathogens transmitted by ticks are an emerging problem worldwide, this study aimed to diagnose the causal agents of infection in dogs presenting suspected hemoparasitoses. Fifty-eight dogs with clinical signs such as depression, hemorrhagic diathesis and fever were evaluated regarding clinical presentation, hemogram, blood smears and serological tests, using the indirect immunofluorescence method for the agents Babesia vogeli and Ehrlichia canis and conventional PCR for Babesia spp. (gene 18S rRNA, Rangelia vitalii (gene 18S rRNA and Ehrlichia spp. (gene dsb. Five (8.6% of the 58 dogs were serologically positive for Babesia spp. and three (5.1% for E. canis. Four dogs (6.8% were positive for R. vitalii through the molecular diagnosis. The PCR products were sequenced and the DNA from R. vitalii was found to be 99% genetically identical to samples of R. vitalii that had been isolated in Brazil. No presence of Babesia spp. or E. canis was observed through PCR on the dogs evaluated here. The results indicate the presence of R. vitalii and exposure to Babesia spp. and Ehrlichia spp. among the dogs analyzed.
Full Text Available In recent years haemosporidian infection by protozoa of the genus Plasmodium and Haemoproteus, has been considered one of the most important factors related to the extinction and/or population decline of several species of birds worldwide. In Brazil, despite the large avian biodiversity, few studies have been designed to detect this infection, especially among wild birds in captivity. Thus, the objective of this study was to analyze the prevalence of Plasmodium spp. and Haemoproteus spp. infection in wild birds in captivity in the Atlantic Forest of southeastern Brazil using microscopy and the polymerase chain reaction. Blood samples of 119 different species of birds kept in captivity at IBAMA during the period of July 2011 to July 2012 were collected. The parasite density was determined based only on readings of blood smears by light microscopy. The mean prevalence of Plasmodium spp. and Haemoproteus spp. infection obtained through the microscopic examination of blood smears and PCR were similar (83.19% and 81.3%, respectively, with Caracara plancus and Saltator similis being the most parasitized. The mean parasitemia determined by the microscopic counting of evolutionary forms of Plasmodium spp. and Haemoproteus spp. was 1.51%. The results obtained from this study reinforce the importance of the handling of captive birds, especially when they will be reintroduced into the wild.
Talaga-Ćwiertnia, Katarzyna; Bulanda, Małgorzata
Vancomycin-resistant Enterococcus faecium (VREfm) strains have become an important hospital pathogen due to their rapid spread, high mortality rate associated with infections and limited therapeutic options. Vancomycin resistance is predominantly mediated by VanA or VanB phenotypes, which differ as regards maintaining sensitivity to teicoplanin in the VanB phenotype. The majority of VREfm cases in the United States, Europe, Korea, South America and Africa are currently caused by the VanA phenotype. However, the epidemics in Australia and Singapore are chiefly brought about by the VanB phenotype. The rate of VREfm isolate spread varies greatly. The greatest percentage of VREfm is now recorded in the USA, Ireland and Australia. Supervision of VRE is implemented to varying degrees. Therefore, the epidemiological situation in some countries is difficult to assess due to limited data or lack thereof.
Singh, Kavindra V.; La Rosa, Sabina Leanti; Somarajan, Sudha R.; Roh, Jung Hyeob
EfbA is a PavA-like fibronectin adhesin of Enterococcus faecalis previously shown to be important in experimental urinary tract infection. Here, we expressed and purified the E. faecalis OG1RF EfbA and confirmed that this protein binds with high affinity to immobilized fibronectin, collagen I, and collagen V. We constructed an efbA deletion mutant and demonstrated that its virulence was significantly attenuated (P endocarditis model. Furthermore, efbA deletion resulted in diminished ability to bind fibronectin (P endocarditis (P = 0.008 versus control). Taken together, our results demonstrate that EfbA is an important factor involved in E. faecalis endocarditis and that rEfbA immunization is effective in preventing such infection, likely by interfering with bacterial adherence. PMID:26351286
Wegener, Henrik Caspar; Aarestrup, Frank Møller; Jensen, Lars Bogø
on the Tn1546 transposon. Furthermore, glycopeptide-resistant strains, as well as resistance determinants, can be transmitted from animals to humans. Two antimicrobial classes expected to provide the future therapeutic options for treatment of infections with vancomycin-resistant enterococci have analogues......Supplementing animal feed with antimicrobial agents to enhance growth has been common practice for more than 30 years and is estimated to constitute more than half the total antimicrobial use worldwide. The potential public health consequences of this use have been debated; however, until recently......, clear evidence of a health risk was not available. Accumulating evidence now indicates that the use of the glycopeptide avoparcin as a growth promoter has created in food animals a major reservoir of Enterococcus faecium, which contains the high level glycopeptide resistance determinant vanA, located...
Gorduysus, Melahat; Nagas, Emre; Torun, Ozgur Yildirim; Gorduysus, Omer
The aim of this study was to compare the in vitro reduction of a bacterial population in a root canal by mechanical instrumentation using three rotary systems and hand instrumentation technique. The root canals contaminated with a suspension of Enterococcus faecalis were instrumented using ProTaper, K3, HeroShaper and K-file hand instrumentation technique. Later the root canals were sampled. After serial dilutions, samples were incubated in culture media for 24 h. Bacterial colonies were counted and the results were given as number of colony-forming units per millilitre. The results showed that all the canal instrumentation systems reduced the number of bacterial cells in the root canals. Statistically, ProTaper instruments were more effective in reducing the number of bacteria than the other rotary files or hand instruments. © 2010 The Authors. Australian Endodontic Journal © 2010 Australian Society of Endodontology.
Sadeghifard, Nourkhoda; Soheili, Sara; Sekawi, Zamberi; Ghafourian, Sobhan
The current study was conducted to investigate the relationship between vancomycin-resistant Enterococcus faecalis (VRE) and the presence of mazEF toxin-antitoxin (TA) system, which may be useful as target for novel antimicrobial therapy concepts. The susceptibility of E. faecalis was determined by MIC, and the presence of the mazEF TA system was evaluated by PCR. Among 200 E. faecalis isolates 39.5% showed resistance to vancomycin (VRE), while 60.5% were susceptible strains (VSE). The mazEF TA system was positive in all VRE isolates (100%), but less prevalent (38/121, 31.4%) among the 121 VSE strains. In conclusion, our study demonstrated a positive relationship between the presence of vancomycin resistance and mazEF TA system. This observation may introduce therapeutic options against a novel antimicrobial target in enterococci.
Izquierdo, Esther; Wagner, Camille; Marchioni, Eric; Aoude-Werner, Dalal; Ennahar, Saïd
Enterococcus faecalis WHE 96, a strain isolated from soft cheese based on its anti-Listeria activity, produced a 5,494-Da bacteriocin that was purified to homogeneity by ultrafiltration and cation-exchange and reversed-phase chromatographies. The amino acid sequence of this bacteriocin, named enterocin 96, was determined by Edman degradation, and its structural gene was sequenced, revealing a double-glycine leader peptide. After a comparison with other bacteriocins, it was shown that enterocin 96 was a new class II bacteriocin that showed very little similarity with known structures. Enterocin 96 was indeed a new bacteriocin belonging to class II bacteriocins. The activity spectrum of enterocin 96 covered a wide range of bacteria, with strong activity against most gram-positive strains but very little or no activity against gram-negative strains. PMID:19411428
Mareková, Mária; Lauková, Andrea; Skaugen, Morten; Nes, Ingolf
The new bacteriocin, termed enterocin M, produced by Enterococcus faecium AL 41 showed a wide spectrum of inhibitory activity against the indicator organisms from different sources. It was purified by (NH4)2SO4 precipitation, cation-exchange chromatography and reverse phase chromatography (FPLC). The purified peptide was sequenced by N-terminal amino acid Edman degradation and a mass spectrometry analysis was performed. By combining the data obtained from amino acid sequence (39 N-terminal amino acid residues was determined) and the molecular weight (determined to be 4628 Da) it was concluded that the purified enterocin M is a new bacteriocin, which is very similar to enterocin P. However, its molecular weight is different from enterocin P (4701.25). Of the first 39 N-terminal residues of enterocin M, valine was found in position 20 and a lysine in position 35, while enterocin P has tryptophane residues in these positions.
Izquierdo, Esther; Wagner, Camille; Marchioni, Eric; Aoude-Werner, Dalal; Ennahar, Saïd
Enterococcus faecalis WHE 96, a strain isolated from soft cheese based on its anti-Listeria activity, produced a 5,494-Da bacteriocin that was purified to homogeneity by ultrafiltration and cation-exchange and reversed-phase chromatographies. The amino acid sequence of this bacteriocin, named enterocin 96, was determined by Edman degradation, and its structural gene was sequenced, revealing a double-glycine leader peptide. After a comparison with other bacteriocins, it was shown that enterocin 96 was a new class II bacteriocin that showed very little similarity with known structures. Enterocin 96 was indeed a new bacteriocin belonging to class II bacteriocins. The activity spectrum of enterocin 96 covered a wide range of bacteria, with strong activity against most gram-positive strains but very little or no activity against gram-negative strains.
Safari, Reza; Adel, Milad; Lazado, Carlo Cabacang
The present study evaluated the benefits of dietary administration of host-derived candidate probiotics Enterococcus casseliflavus in juvenile rainbow trout Oncorhynchus mykiss. Experimental diets were prepared by incorporating the microorganisms in the basal feed at 3 inclusion levels (i.e. 107...... CFU g-1 of feed [T1], 108 CFU g-1 of feed [T2], 109 CFU g-1 of feed [T3]). The probiotic feeds were administered for 8 weeks, with a group fed with the basal diet serving as control. The effects on growth performance, gut health, innate immunity and disease resistance were evaluated.Results showed...... that growth performance parameters were significantly improved in T2 and T3 groups. Activities of digestive enzymes such as trypsin and lipase were significantly higher in these two groups as well. Gut micro-ecology was influenced by probiotic feeding as shown by the significant increase in intestinal lactic...
Strickertsson, Jesper A. B; Desler, Claus; Martin-Bertelsen, Tomas
Background Achlorhydria caused by e.g. atrophic gastritis allows for bacterial overgrowth, which induces chronic inflammation and damage to the mucosal cells of infected individuals driving gastric malignancies and cancer. Enterococcus faecalis (E. faecalis) can colonize achlohydric stomachs and we...... therefore wanted to study the impact of E. faecalis infection on inflammatory response, reactive oxygen species (ROS) formation, mitochondrial respiration, and mitochondrial genetic stability in gastric mucosal cells. Methods To separate the changes induced by bacteria from those of the inflammatory cells...... we established an in vitro E. faecalis infection model system using the gastric carcinoma cell line MKN74. Total ROS and superoxide was measured by fluorescence microscopy. Cellular oxygen consumption was characterized non-invasively using XF24 microplate based respirometry. Gene expression...
Suneel Kumar Chinni
Full Text Available Aim: To determine the antibacterial efficacy of Nisin in comparison with Calcium hydroxide and Chlorhexidine. Materials and Methods: Fifty extracted lower premolar single-rooted teeth were collected and were instrumented with K3 rotary files. Then, root canals were inoculated with a bacterial solution of Enterococcus faecalis. After 21 days, the canals were inoculated with Saline, Nisin, Vancomycin, Calcium hydroxide, and Chlorhexidine. The roots were left for 7 days and on the 8 th day, to investigate the degree of infection of the radicular dentin, specimens of the dentin chips from the full length of the root canal were harvested using a sterile rotary K3 instrument (size 25 6% taper. Results: The results of the present study showed that Nisin and Chlorhexidine showed none of Colony Forming Units (CFU in their respective group. Conclusion: Within the limits of the study, Nisin was effective in eradicating E. faecalis cells in pure culture and root canal dentin.
Marinalda Anselmo Vilela
Full Text Available The isolation of vancomycin resistant enterococci (VRE in Brazil has rapidly increased, following the world wide tendency. We report in the present study the first isolation of vancomycin resistant Enterococcus faecalis (VRE in the Northeast of Brazil. The four VRE isolates were characterized for antimicrobial susceptibility, genotypic typing by macro restriction of chromosomal DNA followed by pulsed-field gel electrophoresis and for characterization of the Tn1546-like element and plasmid contents. The isolates showed resistance to multiple antibiotics and a single genotype profile, suggesting the dissemination of a single clone among the patients. Tn1546 associated to genetic elements as plasmids shows the importance of infection control measures to avoid the spreading of glycopetide resistance by conjugative transfer of VanA elements.
Full Text Available Background and Aims: Instrumentation of the root canals results in formation of smear layer which covers the dentinal tubules. In infected teeth, it is ideal to achieve a material that has the ability to remove the smear layer besides antimicrobial activity. Therefore, this study was designed to evaluate the antimicrobial effect of Citrus aurantifolia extracts (lime juice and rind extract on Enterococcus faecalis within dentinal tubules in the presence of smear layer.Materials and Methods: One-hundred and forty dentin tubes were prepared from bovine incisors. After removal the smear layer, the specimens were infected with Enterococcus faecalis. Then, the smear layer was reformed. Test solutions were used as the irrigants in study roups as follows: group 1: 5.25% NaOCl; group 2: 17% EDTA; group 3: NaOCl+EDTA; group 4: Lime juice; group 5: ethanolic rind extract of C.aurantifolia; group 6: 96% ethanol. Dentin chips were collected from inner and outer layers of dentinal walls and optical density was measured. The data were analyzed using one-way ANOVA and Tamhane tests.Results: In outer layer of dentin, the efficacy of rind extract was less than that of NaOCl+EDTA (P<0.05. Also Lime juice was less effective than EDTA, NaOCl and NaOCl+EDTA (P<0.05. In inner layer of dentin, Lime juice was significantly less effective than NaOCl and NaOCl+EDTA (P<0.05. The efficacy of rind extract was less than that of NaOCl+EDTA (P<0.05.Conclusion: In the presence of smear layer, the antimicrobial activity of Lime juice was less than that of NaOCl but the efficacy of rind extract was similar to that of NaOCl.
Ben Braïek, Olfa; Ghomrassi, Hamdi; Cremonesi, Paola; Morandi, Stefano; Fleury, Yannick; Le Chevalier, Patrick; Hani, Khaled; Bel Hadj, Omrane; Ghrairi, Taoufik
Screening for lactic acid bacteria (LAB) from fresh shrimp samples (Penaeus vannamei) collected from retail seafood markets in the Tunisian's coast, resulted in the isolation of an Enterococcus strain termed Q1. This strain was selected for its antagonistic activity against pathogenic bacteria such as Listeria monocytogenes, Pseudomonas aeruginosa, Lactococcus garvieae and against fungi (Aspergillus niger and Fusarium equiseti). The Q1 strain was characterised using standard morphological and biochemical tests, growth assays at different temperatures, pH and salinity. 16S rRNA, rpoA and pheS gene sequencing, as well as the 16S-23S rRNA intergenic spacer analyses, were combined to identify strain Q1 as a strain of Enterococcus lactis. The bacteriocin produced by E. lactis Q1 is thermostable, active in the pH range from 4.0 to 9.0 and has a bactericidal mode of action. The enterocin P structural gene was detected by specific PCR in strain E. lactis Q1, which is in good agreement with SDS-PAGE data of the purified bacteriocin. A lack of significant antibiotic resistance genes and virulence determinants was confirmed by specific PCRs. This work provides the first description of an enterocin P producer E. lactis strain isolated from a fresh shrimp. Based on its safety properties (absence of haemolytic activity, virulence factors and antibiotic resistance genes), this strain has the potential to be used as a natural additive or adjunct protective culture in food biopreservation and/or probiotic culture.
Fazal, Nadeem; Shelip, Alla; Siddiqui, Erum; Ali, Ashraf; Azim, Anser C; Al-Ghoul, Walid M
Recently we found that superimposition of Enterococcus faecalis infection on burn injury caused an eruption of host mortality not seen with either individual challenge. We hypothesized that the Enterococcus bacteria, and/or factors related to these organisms, aggravate burn-induced modulations in host defense by neutrophils. Our study focuses on alterations in neutrophils' oxidative, proteolytic, and adhesive functions and transendothelial migration of neutrophils in burn rats inoculated with E. faecalis. Rats were subjected to burn (30% total body surface area) and then intra-abdominally inoculated with E. faecalis (10(4)CFU kg(-1) b.w). Polymorphonuclear neutrophils (PMNs) were harvested from circulating/blood and tissue/peritoneal cavity at day-2 post injury. Extracellular release of O(-)(2) anion production was determined by luminometry, and intracellular production of reactive oxygen species was measured by digital imaging technique. Fluoroscan analysis and confocal microscopy determined intracellular elastase production. The expression of adhesion molecule CD11b/CD18 was performed by flow cytometry. Calcein AM-labeled PMNs were co-cultured with TNF-α-stimulated rat lung microvascular endothelial cells, and their ability to adhere was assessed by fluorometry and digital imaging and finally, chemotaxis was measured by neutrophil transmigration assays. The results showed differential effector responses by circulatory and/or tissue PMNs. Tissue/peritoneal PMNs produced more O(-)(2), less intracellular elastase, and increased expression of CD11b/CD18 accompanied with increased adhesivity of MIP-2-stimulated PMNs to endothelial cells as compared to circulatory/blood PMNs. This differential effect was more pronounced following burn plus E. faecalis infection, indicating that the combined injury changed neutrophil functions. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.
Svetoslav D. Todorov
Full Text Available Boza is a low-pH and low-alcohol cereal-based beverage produced in the Balkan Peninsula. Barley was cooked and prepared according to a traditional recipe and inoculated with Enterococcus mundtii ST4V (a potential probiotic and bacteriocin-producing strain, commercially produced boza, Saccharomyces cerevisiae, and a combination of strain E. mundtii ST4V and Saccharomyces cerevisiae. Fermentation was carried out at 37 °C for 3 h. The organoleptic properties of fermented products were evaluated by a qualified taste panel. No significant differences in rheological properties were observed, suggesting that E. mundtii ST4V had no effect on the quality of the final product. Microbial cell numbers remained relatively unchanged during one week of storage. The preservative properties of bacteriocin ST4V were evaluated by contaminating boza with Lactobacillus sakei DSM 20017. Changes in microbial populations were monitored by using classical microbiological methods, PCR with species-specific primers and denaturing gradient gel electrophoresis (DGGE. Adsorption of bacteriocin ST4V to target cells is pH-dependent, with the highest adsorption (88 % recorded at pH=8.0 and pH=10.0. Maximum adsorption of bacteriocin ST4V (75 % to Enterococcus faecalis and Listeria innocua was recorded at 25 to 37 °C. Growth of E. mundtii ST4V was inhibited only by a few antibiotics and anti-inflammatory medicaments, suggesting that the strain may be used as a probiotic by individuals receiving medical treatment.
Full Text Available Ash (Fraxinus spp. is a dominant tree species throughout urban and forested landscapes of North America (NA. The rapid invasion of NA by emerald ash borer (Agrilus planipennis, a wood-boring beetle endemic to Eastern Asia, has resulted in the death of millions of ash trees and threatens billions more. Larvae feed primarily on phloem tissue, which girdles and kills the tree. While NA ash species including black (F. nigra, green (F. pennsylvannica and white (F. americana are highly susceptible, the Asian species Manchurian ash (F. mandshurica is resistant to A. planipennis perhaps due to their co-evolutionary history. Little is known about the molecular genetics of ash. Hence, we undertook a functional genomics approach to identify the repertoire of genes expressed in ash phloem.Using 454 pyrosequencing we obtained 58,673 high quality ash sequences from pooled phloem samples of green, white, black, blue and Manchurian ash. Intriguingly, 45% of the deduced proteins were not significantly similar to any sequences in the GenBank non-redundant database. KEGG analysis of the ash sequences revealed a high occurrence of defense related genes. Expression analysis of early regulators potentially involved in plant defense (i.e. transcription factors, calcium dependent protein kinases and a lipoxygenase 3 revealed higher mRNA levels in resistant ash compared to susceptible ash species. Lastly, we predicted a total of 1,272 single nucleotide polymorphisms and 980 microsatellite loci, among which seven microsatellite loci showed polymorphism between different ash species.The current transcriptomic data provide an invaluable resource for understanding the genetic make-up of ash phloem, the target tissue of A. planipennis. These data along with future functional studies could lead to the identification/characterization of defense genes involved in resistance of ash to A. planipennis, and in future ash breeding programs for marker development.
Pfister, J A; Gardner, D R; Panter, K E; Manners, G D; Ralphs, M H; Stegelmeier, B L; Schoch, T K
Larkspurs (Delphinium spp.) are toxic plants that contain numerous diterpenoid alkaloids which occur as one of two structural types: (1) lycotonine, and (2) 7,8-methylenedioxylycoctonine (MDL-type). Among the lycoctonine type alkaloids are three N-(methylsuccinimido) anthranoyllycoctonine (MSAL-type) alkaloids which appear to be most toxic: methyllycaconitine (MLA), 14-deacetylnudicauline (DAN), and nudicauline. An ester function at C-18 is an important structural requirement for toxicity. Intoxication results from neuromuscular paralysis, as nicotinic acetylcholine receptors in the muscle and brain are blocked by toxic alkaloids. Clinical signs include labored breathing, rapid and irregular heartbeat, muscular weakness, and collapse. Toxic alkaloid concentration generally declines in tall larkspurs with maturation, but alkaloid concentration varies over years and from plant to plant, and is of little use for predicting consumption by cattle. Knowledge of toxic alkaloid concentration is valuable for management purposes when cattle begin to eat larkspur. Cattle generally begin consuming tall larkspur after flowering racemes are elongated, and consumption increases as larkspur matures. Weather is also a major factor in cattle consumption, as cattle tend to eat more larkspur during or just after summer storms. Management options that may be useful for livestock producers include conditioning cattle to avoid larkspur (food aversion learning), grazing tall larkspur ranges before flowering (early grazing) and after seed shatter (late grazing), grazing sheep before cattle, herbicidal control of larkspur plants, and drug therapy for intoxicated animals. Some potentially fruitful research avenues include examining alkaloid chemistry in low and plains larkspurs, developing immunologic methods for analyzing larkspur alkaloids, developing drug therapy, and devising grazing regimes specifically for low and plains larkspur.
Hagen, Ralf Matthias; Frickmann, Hagen; Ehlers, Julian; Krüger, Andreas; Margos, Gabriele; Hizo-Teufel, Cecilia; Fingerle, Volker; Rakotozandrindrainy, Raphael; Kalckreuth, Vera von; Im, Justin; Pak, Gi Deok; Jeon, Hyon Jin; Rakotondrainiarivelo, Jean Philibert; Heriniaina, Jean Noël; Razafindrabe, Tsiry; Konings, Frank; May, Jürgen; Hogan, Benedikt; Ganzhorn, Jörg; Panzner, Ursula; Schwarz, Norbert Georg; Dekker, Denise; Marks, Florian; Poppert, Sven
The occurrence of tick-borne relapsing fever and leptospirosis in humans in Madagascar remains unclear despite the presence of their potential vectors and reservoir hosts. We screened 255 Amblyomma variegatum ticks and 148 Rhipicephalus microplus ticks from Zebu cattle in Madagascar for Borrelia-specific DNA. Borrelia spp. DNA was detected in 21 Amblyomma variegatum ticks and 2 Rhipicephalus microplus ticks. One Borrelia found in one Rhipicephalus microplus showed close relationship to Borrelia theileri based on genetic distance and phylogenetic analyses on 16S rRNA and flaB sequences. The borreliae from Amblyomma variegatum could not be identified due to very low quantities of present DNA reflected by high cycle threshold values in real-time-PCR. It is uncertain whether these low numbers of Borrelia spp. are sufficient for transmission of infection from ticks to humans. In order to determine whether spirochaete infections are relevant in humans, blood samples of 1009 patients from the highlands of Madagascar with fever of unknown origin were screened for Borrelia spp. - and in addition for Leptospira spp. - by real-time PCR. No target DNA was detected, indicating a limited relevance of these pathogens for humans in the highlands of Madagascar. Copyright © 2017 Elsevier B.V. All rights reserved.
Lu, J; Struewing, I; Vereen, E; Kirby, A E; Levy, K; Moe, C; Ashbolt, N
This study investigated waterborne opportunistic pathogens (OPs) including potential hosts, and evaluated the use of Legionella spp. for indicating microbial water quality for OPs within a full-scale operating drinking water distribution system (DWDS). To investigate the occurrence of specific microbial pathogens within a major city DWDS we examined large volume (90 l drinking water) ultrafiltration (UF) concentrates collected from six sites between February, 2012 and June, 2013. The detection frequency and concentration estimates by qPCR were: Legionella spp. (57%/85 cell equivalent, CE l(-1) ), Mycobacterium spp. (88%/324 CE l(-1) ), Pseudomonas aeruginosa (24%/2 CE l(-1) ), Vermamoeba vermiformis (24%/2 CE l(-1) ) and Acanthamoeba spp. (42%/5 cyst equivalent, CE l(-1) ). There was no detection of the following microorganisms: human faecal indicator Bacteroides (HF183), Salmonella enterica, Campylobacter spp., Escherichia coli O157:H7, Giardia intestinalis, Cryptosporidium spp. or Naegleria fowleri. There were significant correlations between the qPCR signals of Legionella spp. and Mycobacterium spp., and their potential hosts V. vermiformis and Acanthamoeba spp. Sequencing of Legionella spp. demonstrated limited diversity, with most sequences coming from two dominant groups, of which the larger dominant group was an unidentified species. Other known species including Legionella pneumophila were detected, but at low frequency. The densities of Legionella spp. and Mycobacterium spp. were generally higher (17 and 324 folds, respectively) for distal sites relative to the entry point to the DWDS. Legionella spp. occurred, had significant growth and were strongly associated with free-living amoebae (FLA) and Mycobacterium spp., suggesting that Legionella spp. could provide a useful DWDS monitoring role to indicate potential conditions for non-faecal OPs. The results provide insight into microbial pathogen detection that may aid in the monitoring of microbial water
Duplan, Florent; Davies, Saran; Filler, Serina; Abdullah, Swaid; Keyte, Sophie; Newbury, Hannah; Helps, Chris R; Wall, Richard; Tasker, Séverine
Ticks derived from cats have rarely been evaluated for the presence of pathogens. The aim of this study was to determine the prevalence of Anaplasma phagocytophilum, Bartonella spp., haemoplasma species and Hepatozoon spp. in ticks collected from cats in the UK. Five hundred and forty DNA samples extracted from 540 ticks collected from cats presenting to veterinarians in UK practices were used. Samples underwent a conventional generic PCR assay for detection of Hepatozoon spp. and real-time quantitative PCR assays for detection of Anaplasma phagocytophilum and three feline haemoplasma species and a generic qPCR for detection of Bartonella spp. Feline 28S rDNA served as an endogenous internal PCR control and was assessed within the haemoplasma qPCR assays. Samples positive on the conventional and quantitative generic PCRs were submitted for DNA sequencing for species identification. Feline 28S rDNA was amplified from 475 of the 540 (88.0%) ticks. No evidence of PCR inhibition was found using an internal amplification control. Of 540 ticks, 19 (3.5%) contained DNA from one of the tick-borne pathogens evaluated. Pathogens detected were: A. phagocytophilum (n = 5; 0.9%), Bartonella spp. (n = 7; 1.3%) [including Bartonella henselae (n = 3; 0.6%) and Bartonella clarridgeiae (n = 1; 0.2%)], haemoplasma species (n = 5; 0.9%), "Candidatus Mycoplasma haemominutum" (n = 3; 0.6%), Mycoplasma haemofelis (n = 1; 0.2%), "Candidatus Mycoplasma turicensis" (n = 1; 0.2%), Hepatozoon spp. (n = 2; 0.4%), Hepatozoon felis (n = 1; 0.2%) and Hepatozoon silvestris (n = 1; 0.2%). These data provide important information on the prevalence of tick-borne pathogens in ticks infesting cats, with the identification of haemoplasma species, A. phagocytophilum, H. felis and Bartonella spp. (including B. henselae and B. clarridgeiae). This study also documents the first report of H. silvestris in ticks collected from domestic cats.
del Socorro Santos Díaz, María; Barba de la Rosa, Ana-Paulina; Héliès-Toussaint, Cécile; Guéraud, Françoise
Opuntia species have been used for centuries as food resources and in traditional folk medicine for their nutritional properties and their benefit in chronic diseases, particularly diabetes, obesity, cardiovascular diseases, and cancer. These plants are largely distributed in America, Africa, and the Mediterranean basin. Opuntia spp. have great economic potential because they grow in arid and desert areas, and O. ficus-indica, the domesticated O. species, is used as a nutritional and pharmaceutical agent in various dietary and value-added products. Though differences in the phytochemical composition exist between wild and domesticated (O. ficus-indica) Opuntia spp., all Opuntia vegetatives (pear, roots, cladodes, seeds, and juice) exhibit beneficial properties mainly resulting from their high content in antioxidants (flavonoids, ascorbate), pigments (carotenoids, betalains), and phenolic acids. Other phytochemical components (biopeptides, soluble fibers) have been characterized and contribute to the medicinal properties of Opuntia spp. The biological properties of Opuntia spp. have been investigated on cellular and animal models and in clinical trials in humans, allowing characterization and clarification of the protective effect of Opuntia-enriched diets in chronic diseases. This review is an update on the phytochemical composition and biological properties of Opuntia spp. and their potential interest in medicine. PMID:28491239
María del Socorro Santos Díaz
Full Text Available Opuntia species have been used for centuries as food resources and in traditional folk medicine for their nutritional properties and their benefit in chronic diseases, particularly diabetes, obesity, cardiovascular diseases, and cancer. These plants are largely distributed in America, Africa, and the Mediterranean basin. Opuntia spp. have great economic potential because they grow in arid and desert areas, and O. ficus-indica, the domesticated O. species, is used as a nutritional and pharmaceutical agent in various dietary and value-added products. Though differences in the phytochemical composition exist between wild and domesticated (O. ficus-indica Opuntia spp., all Opuntia vegetatives (pear, roots, cladodes, seeds, and juice exhibit beneficial properties mainly resulting from their high content in antioxidants (flavonoids, ascorbate, pigments (carotenoids, betalains, and phenolic acids. Other phytochemical components (biopeptides, soluble fibers have been characterized and contribute to the medicinal properties of Opuntia spp. The biological properties of Opuntia spp. have been investigated on cellular and animal models and in clinical trials in humans, allowing characterization and clarification of the protective effect of Opuntia-enriched diets in chronic diseases. This review is an update on the phytochemical composition and biological properties of Opuntia spp. and their potential interest in medicine.
Del Socorro Santos Díaz, María; Barba de la Rosa, Ana-Paulina; Héliès-Toussaint, Cécile; Guéraud, Françoise; Nègre-Salvayre, Anne
Opuntia species have been used for centuries as food resources and in traditional folk medicine for their nutritional properties and their benefit in chronic diseases, particularly diabetes, obesity, cardiovascular diseases, and cancer. These plants are largely distributed in America, Africa, and the Mediterranean basin. Opuntia spp. have great economic potential because they grow in arid and desert areas, and O. ficus-indica , the domesticated O . species, is used as a nutritional and pharmaceutical agent in various dietary and value-added products. Though differences in the phytochemical composition exist between wild and domesticated ( O. ficus-indica ) Opuntia spp., all Opuntia vegetatives (pear, roots, cladodes, seeds, and juice) exhibit beneficial properties mainly resulting from their high content in antioxidants (flavonoids, ascorbate), pigments (carotenoids, betalains), and phenolic acids. Other phytochemical components (biopeptides, soluble fibers) have been characterized and contribute to the medicinal properties of Opuntia spp. The biological properties of Opuntia spp. have been investigated on cellular and animal models and in clinical trials in humans, allowing characterization and clarification of the protective effect of Opuntia -enriched diets in chronic diseases. This review is an update on the phytochemical composition and biological properties of Opuntia spp. and their potential interest in medicine.
Full Text Available Campylobacter spp. is a common cause of bacterial food-borne illness. Birds, especially poultry are primary reservoirs of C. jejuni. The aim of this study was to evaluate the occurrence of Campylobacter spp. in chicken cuts purchased in supermarkets of Londrina, Parana. A total of 50 samples of chicken cuts, such as breasts, thighs and drumsticks were analyzed. The confirmation of the presence of Campylobacter spp. was performed by identifying the suspected colonies on the selective medium using the polymerase chain reaction. Of the 50 samples analyzed, 28 (56% were positive for Campylobacter spp. Chicken meat, as observed in this study, is a possible source of Campylobacter transmission to humans. This study alerts for the importance to analyze the occurrence of Campylobacter in chicken meat, due to the significant number of positive samples observed and no available epidemiological data in Brazil. The correct orientation about handling and cooking of chicken meat is also necessary to prevent human infection by Campylobacter spp.
Grimaldi, A; Bartowsky, E; Jiranek, V
To assess glycosidase activities from a range of Lactobacillus and Pediococcus species and characterize these activities under conditions pertinent to the wine industry. Lactic acid bacteria were cultured in MRS broth supplemented with apple juice before being harvested, washed and assayed for glycosidase activity using p-nitrophenol-linked substrates. All strains exhibited a detectable capacity for the hydrolysis of the beta- and alpha-d-glucopyranosides. The magnitude of these activities and their response to the physico-chemical parameters investigated varied in a strain-dependent manner. The use of an assay buffer with a pH below 4 generally resulted in a reduced hydrolysis of both substrates while temperature optima ranged between 35 and 45 degrees C. The effect of the inclusion of ethanol in the assay buffer (up to 12%, v/v) ranged from near complete inhibition to increases in activity approaching 80%. With the clear exception of a single strain, glucose and fructose (0.1-20 g l(-1)) acted as inhibitors. An assessment of glycosidase activity during simultaneous exposure to glucose and ethanol at a pH of 3.5 suggested that ethanol decreased loss of activity under these wine-like conditions. Lactobacillus spp. and Pediococcus spp. possess varying degrees of beta- and alpha-d-glucopyranosidase activities, which in turn are influenced differently by exposure to ethanol and/or sugars, temperature and pH. Several strains appeared suited for further evaluation under winemaking conditions. This work highlights the fact that strains of Lactobacillus and Pediococcus have the potential to influence the glycoside composition of wine. Tailoring of wine may therefore be possible through selective application of strains or enzymatic extracts thereof.
Nunes, Maria Antónia; Rodrigues, Francisca; Alves, Rita C; Oliveira, Maria Beatriz P P
Herbs have been used from ancient times for infusion preparation based on their potential health effects. In particular, the consumption of Hibiscus sabdariffa L., Crataegus spp. and Panax spp. has been largely associated to cardiovascular benefits. In this work, the label information of 52 herbal products for infusion preparation containing the referred herbs was analyzed and discussed, taking into consideration the European Union regulation for herbal products, which intends to protect public health and harmonize the legal framework in Member States. Details about the cardiovascular-related statements and warning notifications about consumption were considered. Also, regulatory issues and possible herb-drug interactions were explored and discussed. A total of 14 of the 52 herbal products selected presented health claims/statements on the label. Hibiscus was present in the majority of the products and, in some cases, it was mentioned only in the ingredients list and not on the product front-of-pack. Despite the promising outcomes of these plants to modulate cardiovascular risk markers, consumers with some sort of cardiovascular dysfunction and/or under medication treatments should be aware to carefully analyze the labels and consult additional information related to these herbal products. Manufacturers have also a huge responsibility to inform consumers by presenting awareness statements. Lastly, health professionals must advise and alert their patients about possible interactions that could occur between the concomitant consumption of drugs and herbs. Overall, there is still a real need of additional studies and clinical trials to better understand herbs effects and establish a science-based guidance to assess their safety. Copyright © 2017 Elsevier Ltd. All rights reserved.
Atividade antimicrobiana in vitro de quinupristina/dalfopristina para cocos gram-positivos isolados de cinco centros brasileiros: resultado do estudo de vigilância L-SMART Antimicrobial in vitro activity of quinupristin/dalfopristin against gram-positive cocci isolated from 5 Brazilian centers: results from the local smart (L-SMART surveillance study
Full Text Available Há alguns anos tem-se verificado um aumento progressivo da resistência de alguns cocos gram-positivos a determinados antimicrobianos. Este aumento da resistência tem sido observado principalmente no ambiente hospitalar, e as bactérias mais comumente envolvidas são os Staphylococcus spp. e os Enterococcus spp. Devido a este fato, novos antimicrobianos são avaliados para o tratamento de infecções causadas por estas cepas multirresistentes. A associação quinupristina/dalfopristina (Q/D, também conhecida como Synercid®, é um antibacteriano da classe das estreptograminas, de uso endovenoso, composto por dois derivados semi-sintéticos da pristinamicina. A combinação das estreptograminas B e A na razão de 30:70 tem atividade antimicrobiana voltada para cocos gram--positivos, como Staphylococcus spp., Streptococcus spp., incluindo S. pneumoniae e Enterococcus faecium, sendo o E. faecalis habitualmente resistente. Neste estudo foi avaliada a atividade in vitro de Q/D e outros oito antimicrobianos frente a 631 amostras de cocos gram-positivos isoladas de cinco centros brasileiros, complementadas com outras 20 cepas de E. faecium resistentes à vancomicina, provenientes dos Estados Unidos. Para a avaliação da sensibilidade aos antimicrobianos foi determinada a concentração inibitória mínima (MIC pelo método do Etest (AB Biodisk, Solna, Suécia e as cepas testadas foram: Staphylococcus aureus (n = 267, Staphylococcus coagulase negativo (n = 131, Streptococcus pneumoniae (n = 130, Streptococcus beta-hemolíticos (n = 28, Enterococcus faecalis (n = 44 e E. faecium (n = 51. A Q/D demonstrou excelente atividade contra Staphylococcus spp., independente de serem sensíveis ou resistentes à oxacilina. Para S. pneumoniae, a Q/D apresentou igualmente uma ótima atividade, inclusive para as cepas com resistência intermediária ou total para penicilina. Entre as cepas de E. faecium sensíveis à vancomicina, o MIC90 de Q/D obtido foi de 3µg
Full Text Available Our study aimed at using PCR to identify the incidence of Bartonella spp. in blood of dogs. Altogether 286 dogs of 92 breeds aged 3 month to 17 years were tested from October 2008 to December 2009. Healthy dogs as well as dogs with various clinical symptoms of disease were included in the group. Samples were tested by polymerase chain reaction (PCR specific for the presence of Bartonella spp. Following the DNA examination in 286 dogs by PCR and subsequent sequencing, two samples were identified as Bartonella henselae (0.7%. Other species of Bartonella were not found. It was the first time in the Czech Republic when incidence of Bartonella spp. was determined in dogs.
An IRAF package to assist in SPP code development and debugging is described. SPP is the machine-independent programming language used by virtually all IRAF tasks. Tools have been written to aide both novice and advanced SPP programmers with development and debugging by providing tasks to check the code for the number and type of arguments in all calls to IRAF VOS library procedures, list the calling sequences of IRAF tasks, create a database of identifiers for quick access, check for memory which is not freed, and a source code formatter. Debugging is simplified since the programmer is able to get a better understanding of the structure of his/her code, and IRAF library procedure calls (probably the most common source of errors) are automatically checked for correctness.