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1.
Nat Commun ; 12(1): 1523, 2021 03 09.
Artigo em Inglês | MEDLINE | ID: mdl-33750782

RESUMO

Enterococcus faecalis is a commensal and nosocomial pathogen, which is also ubiquitous in animals and insects, representing a classical generalist microorganism. Here, we study E. faecalis isolates ranging from the pre-antibiotic era in 1936 up to 2018, covering a large set of host species including wild birds, mammals, healthy humans, and hospitalised patients. We sequence the bacterial genomes using short- and long-read techniques, and identify multiple extant hospital-associated lineages, with last common ancestors dating back as far as the 19th century. We find a population cohesively connected through homologous recombination, a metabolic flexibility despite a small genome size, and a stable large core genome. Our findings indicate that the apparent hospital adaptations found in hospital-associated E. faecalis lineages likely predate the "modern hospital" era, suggesting selection in another niche, and underlining the generalist nature of this nosocomial pathogen.


Assuntos
Infecção Hospitalar/microbiologia , Enterococcus faecalis/genética , Enterococcus faecalis/metabolismo , Prednisolona/metabolismo , Prednisolona/farmacologia , Animais , Antibacterianos , Aves , Farmacorresistência Bacteriana/genética , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/isolamento & purificação , Genes MDR/genética , Genoma Bacteriano , Infecções por Bactérias Gram-Positivas/microbiologia , Hospitais , Especificidade de Hospedeiro , Humanos , Filogenia , Fatores de Virulência , Sequenciamento Completo do Genoma
2.
Int J Food Microbiol ; 345: 109130, 2021 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-33735781

RESUMO

Pélardon is an artisanal French raw goat's milk cheese, produced using natural whey as a backslop. The aim of this study was to identify key microbial players involved in the acidification and aroma production of this Protected Designation of Origin cheese. Microbial diversity of samples, collected from the raw milk to 3-month cheese ripening, was determined by culture-dependent (MALDI-TOF analysis of 2877 isolates) and -independent (ITS2 and 16S metabarcoding) approaches and linked to changes in biochemical profiles (volatile compounds and acids). In parallel, potential dominant autochthonous microorganism reservoirs were also investigated by sampling the cheese-factory environment. Complex and increasing microbial diversity was observed by both approaches during ripening although major discrepancies were observed regarding Lactococcus lactis and Lacticaseibacillus paracasei fate. By correlating microbial shifts to biochemical changes, Lactococcus lactis was identified as the main acidifying bacterium, while L. mesenteroides and Geotrichum candidum were prevalent and associated with amino acids catabolism after the acidification step. The three species were dominant in the whey (backslop). In contrast, L. paracasei, Enterococcus faecalis, Penicillium commune and Scopulariopsis brevicaulis, which dominated during ripening, likely originated from the cheese-making environment. All these four species were positively correlated to major volatile compounds responsible for the goaty and earthy Pélardon cheese aroma. Overall, this work highlighted the power of MALDI-TOF and molecular techniques combined with volatilome analyses to dynamically follow and identify microbial communities during cheese-making and successively identify the key-players involved in aroma production and contributing to the typicity of Pélardon cheese.


Assuntos
Bactérias/classificação , Bactérias/metabolismo , Queijo/microbiologia , Fungos/classificação , Fungos/metabolismo , Leite/microbiologia , Animais , Bactérias/isolamento & purificação , Enterococcus faecalis/isolamento & purificação , Enterococcus faecalis/metabolismo , Fungos/isolamento & purificação , Geotrichum/isolamento & purificação , Geotrichum/metabolismo , Cabras , Lactobacillus paracasei/isolamento & purificação , Lactobacillus paracasei/metabolismo , Lactococcus lactis/isolamento & purificação , Lactococcus lactis/metabolismo , Microbiota , Odorantes/análise , Penicillium/isolamento & purificação , Penicillium/metabolismo , Scopulariopsis/isolamento & purificação , Scopulariopsis/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
3.
BMJ Case Rep ; 14(1)2021 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-33504526

RESUMO

To report the clinical course and management of interface keratitis due to Enterococcus faecalis after Descemet membrane endothelial keratoplasty (DMEK). A 64-year-old man underwent DMEK, with unevenful immediate postoperative course, with a visual recovery of 20/30 at 2 weeks. At 3 months of clinical visit, interface keratitis was noted. DMEK graft removal with stromal bed scrapings was performed. A diagnosis of E. faecalis interface keratitis was made. The patient responded favourably to antibiotic susceptibility-guided intensive treatment with vancomycin 5% with complete resolution of infection. After 2 months of graft removal, Descemet stripping endothelial keratoplasty (DSEK) was performed. The corneal clarity was restored and the best corrected visual acuity was 20/40 at last follow-up of 1 year. E. faecalis should be kept as a differential in delayed onset interface keratitis after DMEK. After microbiological cure with antibiotic therapy, visual rehabilitation with DSEK restores corneal clarity and results in favourable visual outcome.


Assuntos
Transplante de Córnea , Enterococcus faecalis/isolamento & purificação , Infecções por Bactérias Gram-Positivas/diagnóstico , Ceratite/diagnóstico , Infecção da Ferida Cirúrgica/diagnóstico , Antibacterianos/uso terapêutico , Farmacorresistência Bacteriana Múltipla , Enterococcus faecalis/fisiologia , Infecções por Bactérias Gram-Positivas/tratamento farmacológico , Humanos , Ceratite/tratamento farmacológico , Masculino , Pessoa de Meia-Idade , Infecção da Ferida Cirúrgica/tratamento farmacológico , Tomografia de Coerência Óptica , Vancomicina/uso terapêutico
5.
J Med Microbiol ; 69(9): 1151-1168, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32840477

RESUMO

Introduction. Enterococcus faecalis is a facultative, anaerobic, opportunistic pathogen associated with medical and dental diseases. Bacterial phenotypic traits and pathogenesis are often influenced by lysogeny.Aim. The aim of this study was to characterize both the morphology and complete genome sequences of induced prophages purified from E. faecalis clinical isolates.Methodology. E. faecalis isolates were recovered from the roots of teeth of patients attending an endodontic clinic. The morphological features of isolated phage were characterized using transmission electron microscopy (TEM). DNA sequencing was performed using the Illumina MiSeq platform.Results. TEM indicated that the isolated φEf-vB1 prophage belongs to the family Siphoviridae. The φEf-vB1 prophage was stable over a wide range of temperatures and pH. Sequencing of φEf-vB1 DNA revealed that the phage genome is 37 561 bp in length with a G+C content of 37.6mol% and contained 53 ORFs. Comparison with previously predicted prophage genomes using blast revealed that φEf-vB1 has a high sequence similarity to previously characterized phage genomes. The lysogenic E. faecalis strain exhibited a higher biofilm formation capacity relative to the non-lysogenic strain.Conclusion. The current findings highlight the role of lysogeny in modification of E. faecalis properties and reveal the potential importance of prophages in E. faecalis biology and pathogenesis.


Assuntos
Bacteriófagos/fisiologia , Enterococcus faecalis/fisiologia , Enterococcus faecalis/virologia , Prófagos/fisiologia , Siphoviridae/isolamento & purificação , Composição de Bases , Cavidade Pulpar/microbiologia , Enterococcus faecalis/genética , Enterococcus faecalis/isolamento & purificação , Genoma Viral , Infecções por Bactérias Gram-Positivas/microbiologia , Humanos , Lisogenia , Fases de Leitura Aberta , Periodontite , Prófagos/classificação , Prófagos/genética , Prófagos/isolamento & purificação , Siphoviridae/classificação , Siphoviridae/genética , Siphoviridae/fisiologia
6.
Rev. Asoc. Odontol. Argent ; 108(2): 46-51, mayo-ago. 2020. tab
Artigo em Espanhol | LILACS | ID: biblio-1121108

RESUMO

Objetivos: Comparar ex vivo la eficacia del instrumento XP-endo Finisher y del sistema EndoActivator en la reducción/eliminación del biofilm microbiano en conductos radiculares infectados. Materiales y métodos: Se utilizaron 23 premolares inferiores humanos extraídos cuya longitud fue estandarizada en 17 mm. Todos los conductos se prepararon con el sistema WaveOne Gold Medium (#35.06). Los dientes se esterilizaron, se inocularon con Enterococcus faecalis y se separaron en dos grupos experimentales de 10 piezas cada uno. De los 3 dientes remanentes, 1 fue utilizado como control positivo y 2, como controles negativos. En el grupo 1, las soluciones irrigantes se agitaron con XP-endo Finisher. En el grupo 2, se utilizó EndoActivator. Se tomaron muestras antes de la contaminación, luego de esta y después de la agitación de los irrigantes mediante conos de papel estériles. La carga microbiana fue sembrada en agar sangre y los conos se cultivaron en caldo tripteína de soja. La remoción de la carga microbiana se determinó por la presencia o ausencia de turbiedad del medio. Las unidades formadoras de colonias (UFC) remanentes se cuantificaron y los resultados se categorizaron como R1 (≤10 UFC) o R2 (>10 UFC). Los datos fueron analizados mediante la prueba de Fisher. Resultados: No hubo diferencias significativas entre XP-endo Finisher y EndoActivator (P>0,05). El número de usos no influyó sobre la capacidad operativa de ambos instrumentos (AU)


Aim: To compare ex vivo the effectiveness of the XP-endo Finisher and the EndoActivator in biofilm reduction/ removal from infected root canals. Materials and methods: Twenty three extracted human single-rooted lower premolars were selected and standardised to 17 mm in length. All the canals were prepared with WaveOne Gold Medium reciprocating files (#35.06). The teeth were autoclaved and inoculated with Enterococcus faecalis. The infected teeth were then assigned to 2 experimental groups of 10 teeth each according to the final irrigation/agitation protocol. Of the three remaining teeth, one was used as a positive control, and the other two were used as negative controls. In Group 1 the irrigating solutions were agitated with XP-endo Finisher while in Group 2 the EndoActivator was used. All root canals were sampled before and after contamination, and again after irrigant agitation with sterile paper points. The microbial load was spread on blood agar plates and the paper points were cultured in sterile trypticase soy broth. The removal of the microbial load was determined by visual observation of the turbidity of the media and by quantification of the number of colony-forming units (UFC). The results were categorized as R1 (≤10 UFC) or R2 (>10 UFC). Data were analysed by the Fisher's exact test at P<0.05. Results: No significant differences was found between XP-endo Finisher and EndoActivator (P>0.05) regarding their effectiveness in the reduction/removal of the microbial biofilm. The number of uses of both instruments did not affect their operative performance (AU) Conclusion: XPF and EA were both equally effective for microbial biofilm reduction/removal from ex vivo infected root canals (AU)


Assuntos
Irrigantes do Canal Radicular/química , Equipamentos Odontológicos de Alta Rotação , Biofilmes , Instrumentos Odontológicos , Cavidade Pulpar/microbiologia , Técnicas In Vitro , Contagem de Colônia Microbiana/métodos , Eficácia , Análise Estatística , Enterococcus faecalis/isolamento & purificação , Meios de Cultura
7.
Acta Odontol Latinoam ; 33(1): 45-49, 2020 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-32621599

RESUMO

The gutta-percha cones used in endodontic treatment are produced in aseptic conditions and their composition includes zinc oxide, which is responsible for antibacterial activity. However, there is the possibility of microbial contamination by manipulation, aerosol or during storage. Although several chemical agents have been tested for their decontamination, there is no consensus on the best disinfection protocol to be used. The aim of this study was to evaluate the decontamination of gutta-percha cones contaminated with the bacteria Enterococcus faecalis, by using chlorhexidine digluconate (CHX) and sodium hypochlorite (NaClO) at different concentrations for short exposure times. For this purpose, gutta-percha cones (size 40) were selected at random from a sealed box and immersed for 1 min in a microbial suspension. Then they were immersed in specific Petri dishes for different groups containing: CHX 2%, NaClO 1% or NaClO 2.5% for 30 s or 1 min, and subsequently placed in tubes containing BHI broth. After incubating the tubes for 48 h, it was observed that 1% and 2.5% NaClO and 2% CHX were effective for decontaminating the cones at those exposure time intervals. Microbial growth was detected in one of the replicates of the group with CHX applied for 30 s. To prevent the possibility of failures at this stage, the exposure time of gutta-percha cones to the decontaminating agent should not be reduced.


Assuntos
Clorexidina/análogos & derivados , Descontaminação/métodos , Desinfetantes de Equipamento Odontológico/farmacologia , Enterococcus faecalis/efeitos dos fármacos , Guta-Percha , Irrigantes do Canal Radicular/farmacologia , Hipoclorito de Sódio/farmacologia , Esterilização/métodos , Anti-Infecciosos Locais , Clorexidina/farmacologia , Desinfetantes de Equipamento Odontológico/administração & dosagem , Enterococcus faecalis/isolamento & purificação , Contaminação de Equipamentos/prevenção & controle , Humanos , Materiais Restauradores do Canal Radicular , Irrigantes do Canal Radicular/administração & dosagem , Hipoclorito de Sódio/administração & dosagem
8.
BMC Infect Dis ; 20(1): 356, 2020 May 19.
Artigo em Inglês | MEDLINE | ID: mdl-32517758

RESUMO

BACKGROUND: Vancomycin-resistant Enterococcus spp. (VRE) have spread all over the world. The present study aims to investigate the species distribution, specimen type and susceptibilities of Enterococcal species collected from Nanjing Drum Tower Hospital from 2013 to 2018. Additionally, distribution of VRE and prevalence of van gene among VRE isolates were also analyzed. METHODS: The susceptibilities of 3913 Enterococcus isolates were retrospectively investigated. Among these strains, 60 VRE strains were further anazlyed in this study. The minimum inhibitory concentrations (MICs) of the VRE strains towards vancomycin, teicoplanin and linezolid were determined by E-test. Polymerase chain reaction (PCR) and DNA sequencing were used to investigate the prevalence of van genes among VRE. Furthermore, the sequence types (STs) of VRE strains were explored by multi-locus sequence typing (MLST). RESULTS: Among the 3913 enterococci isolates, Enterococcus faecalis (n = 1870, 47.8%) and Enterococcus faecium (1738, 44.4%) were the main isolates. These Enterococcus strains were mainly isolated from urine (n = 1673, 42.8%), followed by secretions (n = 583, 14.9%) and ascites (n = 554, 14.2%). VRE displayed a decreasing trend year by year. Molecular analysis revealed that 49 out of 60 VRE isolates carried vanA gene, 10 carried vanM, and 1 carried both vanA and vanM genes. Sixteen distinct STs were identified among the 58 VREM, with ST78 (n = 16), ST192 (n = 8) and ST570 (n = 7) being the most dominant ones. CONCLUSIONS: E. faecalis and E. faecium were the major enterococci strains which are the main pathogens of urinary traction infections; vanA and vanM were the main determinants conferring resistance to vancomycin; ST78, ST192 and ST570 were the leading STs of VREM which displayed a decreasing trend of prevalence year by year.


Assuntos
Enterococcus/isolamento & purificação , Infecções por Bactérias Gram-Positivas/microbiologia , Enterococos Resistentes à Vancomicina/isolamento & purificação , Antibacterianos/farmacologia , China , DNA Bacteriano/metabolismo , Enterococcus/efeitos dos fármacos , Enterococcus/genética , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/genética , Enterococcus faecalis/isolamento & purificação , Enterococcus faecium/efeitos dos fármacos , Enterococcus faecium/genética , Enterococcus faecium/isolamento & purificação , Infecções por Bactérias Gram-Positivas/diagnóstico , Humanos , Testes de Sensibilidade Microbiana , Tipagem de Sequências Multilocus , Estudos Retrospectivos , Centros de Atenção Terciária , Vancomicina/farmacologia , Enterococos Resistentes à Vancomicina/efeitos dos fármacos , Enterococos Resistentes à Vancomicina/genética
9.
PLoS One ; 15(4): e0232165, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32343730

RESUMO

We have recently demonstrated that collagenolytic Enterococcus faecalis plays a key and causative role in the pathogenesis of anastomotic leak, an uncommon but potentially lethal complication characterized by disruption of the intestinal wound following segmental removal of the colon (resection) and its reconnection (anastomosis). Here we hypothesized that comparative genetic analysis of E. faecalis isolates present at the anastomotic wound site before and after surgery would shed insight into the mechanisms by which collagenolytic strains are selected for and predominate at sites of anastomotic disruption. Whole genome optical mapping of four pairs of isolates from rat colonic tissue obtained following surgical resection (herein named "pre-op" isolates) and then 6 days later from the anastomotic site (herein named "post-op" isolates) demonstrated that the isolates with higher collagenolytic activity formed a distinct cluster. In order to perform analysis at a deeper level, a single pair of E. faecalis isolates (16A pre-op and 16A post-op) was selected for whole genome sequencing and assembled using a hybrid assembly algorithm. Comparative genomics demonstrated absence of multiple gene clusters, notably a pathogenicity island in the post-op isolate. No differences were found in the fsr-gelE-sprE genes (EF1817-1822) responsible for regulation and production of collagenolytic activity. Analysis of unique genes among the 16A pre-op and post-op isolates revealed the predominance of transporter systems-related genes in the pre-op isolate and phage-related and hydrolytic enzyme-encoding genes in the post-op isolate. Despite genetic differences observed between pre-op and post-op isolates, the precise genetic determinants responsible for their differential expression of collagenolytic activity remains unknown.


Assuntos
Anastomose Cirúrgica , Colo/cirurgia , Enterococcus faecalis/genética , Anastomose Cirúrgica/efeitos adversos , Fístula Anastomótica/etiologia , Fístula Anastomótica/microbiologia , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Mapeamento Cromossômico , Colagenases/genética , Colagenases/metabolismo , Enterococcus faecalis/enzimologia , Enterococcus faecalis/isolamento & purificação , Microbioma Gastrointestinal/genética , Genoma Bacteriano , Intestinos/microbiologia , Ratos , Virulência/genética
10.
J Dairy Sci ; 103(5): 4068-4077, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32197843

RESUMO

The enterococci are ubiquitous bacteria able to colonize the human and animal gastrointestinal tracts and fresh and fermented food products. Their highly plastic genome allows Enterococcus spp. to gain resistance to multiple antibiotics, making infections with these organisms difficult to treat. Food-borne enterococci could be carriers of antibiotic resistance determinants. The goal of this work was to study the characteristics of Enterococcus spp. in fermented milk products from Poland and their antibiotic resistance gene profiles. A total of 189 strains were isolated from 182 dairy products out of 320 samples tested. The predominant species were Enterococcus faecium (53.4%) and Enterococcus faecalis (34.4%). Isolates were resistant to streptomycin (29.1%), erythromycin (14.3%), tetracycline (11.6%), rifampicin (8.7%), and tigecycline (8.1%). We also detected 2 vancomycin-resistant and 3 linezolid-resistant strains; however, no vanA or vanB genes were identified. A total of 57 high-level aminoglycoside resistance strains (30.2%) were identified, most of which have the ant(6')-Ia gene, followed by the aac(6')-Ie-aph(2″)-Ia and aph(3″)-IIIa genes. Resistance to tetracycline was most often conferred by tetM and tetL genes. Macrolide resistance was most frequently encoded by ermB and ermA genes. Conjugative mobile genetic element (transposon Tn916-Tn1545) was identified in 15.3% of the strains, including 96.3% of strains harboring the tetM gene. This study found that enterococci are widely present in retail ready-to-eat dairy products in Poland. Many isolated strains are antibiotic resistant and carry transferable resistance genes, which represent a potential source of transmission of multidrug-resistant bacteria to humans.


Assuntos
Antibacterianos/farmacologia , Laticínios/microbiologia , Farmacorresistência Bacteriana Múltipla , Enterococcus faecalis/isolamento & purificação , Enterococcus faecium/isolamento & purificação , Animais , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecium/efeitos dos fármacos , Microbiologia de Alimentos , Genótipo , Testes de Sensibilidade Microbiana , Polônia
12.
J Am Coll Cardiol ; 75(5): 482-494, 2020 02 11.
Artigo em Inglês | MEDLINE | ID: mdl-32029130

RESUMO

BACKGROUND: Enterococcal endocarditis (EE) is a growing entity in Western countries. However, quality data from large studies is lacking. OBJECTIVES: The purpose of this study was to describe the characteristics and analyze the prognostic factors of EE in the GAMES cohort. METHODS: This was a post hoc analysis of a prospectively collected cohort of patients from 35 Spanish centers from 2008 to 2016. Characteristics and outcomes of 516 cases of EE were compared with those of 3,308 cases of nonenterococcal endocarditis (NEE). Logistic regression and Cox proportional hazards regression analysis were performed to investigate risk factors for in-hospital and 1-year mortality, as well as relapses. RESULTS: Patients with EE were significantly older; more frequently presented chronic lung disease, chronic heart failure, prior endocarditis, and degenerative valve disease; and had higher median age-adjusted Charlson score. EE more frequently involved the aortic valve and prosthesis (64.3% vs. 46.7%; p < 0.001; and 35.9% vs. 28.9%; p = 0.002, respectively) but less frequently pacemakers/defibrillators (1.5% vs. 10.5%; p < 0.001), and showed higher rates of acute heart failure (45% vs. 38.3%; p = 0.005). Cardiac surgery was less frequently performed in EE (40.7% vs. 45.9%; p = 0.024). No differences in in-hospital and 1-year mortality were found, whereas relapses were significantly higher in EE (3.5% vs. 1.7%; p = 0.035). Increasing Charlson score, LogEuroSCORE, acute heart failure, septic shock, and paravalvular complications were risk factors for mortality, whereas prior endocarditis was protective and persistent bacteremia constituted the sole risk factor for relapse. CONCLUSIONS: Besides other baseline and clinical differences, EE more frequently affects prosthetic valves and less frequently pacemakers/defibrillators. EE presents higher rates of relapse than NEE.


Assuntos
Endocardite Bacteriana/epidemiologia , Enterococcus faecalis/isolamento & purificação , Infecções por Bactérias Gram-Positivas/epidemiologia , Idoso , Estudos de Coortes , Endocardite Bacteriana/microbiologia , Feminino , Infecções por Bactérias Gram-Positivas/microbiologia , Humanos , Masculino , Pessoa de Meia-Idade , Espanha/epidemiologia
13.
J Endod ; 46(3): 419-424, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31980201

RESUMO

INTRODUCTION: The aim of this study was to evaluate the effect of using the XP-endo Finisher (XPF; FKG Dentaire, La Chaux de Fonds, Switzerland) in teeth that have a traditional access cavity (TEC) and a contracted access cavity (CEC) design on the amount of decrease in the number of Enterococcus faecalis bacteria within the root canal system. METHODS: Eighty mandibular first molar teeth were selected and randomly divided into 2 groups: TEC and CEC (n = 40/group). After access cavity preparation in the 2 groups, 80 mesiobuccal root canals were contaminated with Enterococcus faecalis for 4 weeks. After the first sampling (S1), in order to perform root canal instrumentation, the TEC and CEC groups were further divided into 4 subgroups (10 teeth/group): Reciproc (VDW GmbH, Munich, Germany) and ProTaper Next (Dentsply Maillefer, Ballaigues, Switzerland) with or without XPF. Bacterial sampling from the root canals was performed with sterile paper points before (S1) and after (S2) instrumentation to determine the bacterial load. The bacterial reduction was counted as colony-forming units/mL and analyzed statistically by 3-factor repeated measures analysis of variance. Multiple comparisons of the main factor effect were performed using the Bonferroni correction (α < .05), all at 5% significance. RESULTS: The number of E. faecalis bacteria in all the samples with different cavity designs were significantly reduced after instrumentation. The lowest value of bacterial decrease percentage was observed in the CEC-Reciproc-XPF (82.8%) group. CONCLUSIONS: The bacterial reduction counts of E. faecalis were a similar level in the TEC and CEC cavities, and the use of XPF did not show significant differences between groups.


Assuntos
Cavidade Pulpar , Enterococcus faecalis , Preparo de Canal Radicular , Instrumentos Odontológicos , Enterococcus faecalis/isolamento & purificação , Alemanha , Preparo de Canal Radicular/métodos
14.
Arch Microbiol ; 202(4): 765-772, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-31822952

RESUMO

This research was conducted using 50 samples of popular traditional cheeses and 160 enterococcal clinical isolates. Phenotypic and genotypic methods used for identification of enterococci. Then, the incidences of antibacterial resistance and virulence traits were investigated. In total, 165 E. faecalis and 43 E. faecium obtained from traditional cheeses and different clinical isolates were analyzed in the study. Antibiotic susceptibility testing revealed 175(84.1%) isolates with multi-drug resistance (MDR) patterns, which was more common among clinical sources. The predominant virulence profile, including gelE, asa1 and cpd was detected within 47 (22.6%) of the MDR isolates. Our results showed that traditional cheeses and clinical E. faecalis isolates have distinct patterns of virulence traits. The identified enterococci with antibiotic resistance and associated virulence factors, could provide a potential risk to the public health.


Assuntos
Queijo/microbiologia , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/patogenicidade , Enterococcus faecium/efeitos dos fármacos , Enterococcus faecium/patogenicidade , Infecções por Bactérias Gram-Positivas/microbiologia , Antibacterianos/farmacologia , Farmacorresistência Bacteriana/efeitos dos fármacos , Enterococcus faecalis/genética , Enterococcus faecalis/isolamento & purificação , Enterococcus faecium/genética , Enterococcus faecium/isolamento & purificação , Genótipo , Humanos , Irã (Geográfico) , Testes de Sensibilidade Microbiana , Fenótipo , Fatores de Virulência/genética
15.
J Dairy Sci ; 103(2): 1223-1237, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31759592

RESUMO

Raw milk contains wide microbial diversity, composed mainly of lactic acid bacteria (LAB), which are used as probiotics in both human and animal husbandry. We isolated, characterized, and evaluated LAB from indigenous Bangladeshi raw milk to assess probiotic potential, including antagonistic activity (against Escherichia coli O157: H7, Enterococcus faecalis, Salmonella Typhimurium, Salmonella Enteritidis, and Listeria monocytogenes), survivability in simulated gastric juice, tolerance to phenol and bile salts, adhesion to ileum epithelial cells, auto- and co-aggregation, hydrophobicity, α-glucosidase inhibitory activity, and antibiotic susceptibility tests. The 4 most promising LAB strains showed probiotic potential and were identified as Lactobacillus casei, Lactobacillus plantarum (which produced plantaricin EF), Lactobacillus fermentum, and Lactobacillus paracasei. These strains inhibited all pathogens tested at various degrees, and competitively excluded pathogens with viable counts of 3.0 to 6.0 log cfu/mL. Bacteriocin, organic acids, and low-molecular-weight substances were mainly responsible for antimicrobial activity by the LAB strains. All 4 LAB strains were resistant to oxacillin and 3 were resistant to vancomycin and streptomycin, with multiple antibiotic resistance indices >0.2. After further in vivo evaluation, these LAB strains could be considered probiotic candidates with application in the food industry.


Assuntos
Lactobacillales/fisiologia , Leite/microbiologia , Probióticos , Animais , Bacteriocinas/metabolismo , Bovinos , Enterococcus faecalis/isolamento & purificação , Enterococcus faecalis/fisiologia , Feminino , Suco Gástrico/microbiologia , Cabras , Humanos , Lactobacillales/isolamento & purificação , Lactobacillus casei/isolamento & purificação , Lactobacillus casei/fisiologia , Lactobacillus fermentum/isolamento & purificação , Lactobacillus fermentum/fisiologia , Lactobacillus plantarum/isolamento & purificação , Lactobacillus plantarum/fisiologia , Probióticos/farmacologia
16.
Nature ; 575(7783): 505-511, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31723265

RESUMO

Chronic liver disease due to alcohol-use disorder contributes markedly to the global burden of disease and mortality1-3. Alcoholic hepatitis is a severe and life-threatening form of alcohol-associated liver disease. The gut microbiota promotes ethanol-induced liver disease in mice4, but little is known about the microbial factors that are responsible for this process. Here we identify cytolysin-a two-subunit exotoxin that is secreted by Enterococcus faecalis5,6-as a cause of hepatocyte death and liver injury. Compared with non-alcoholic individuals or patients with alcohol-use disorder, patients with alcoholic hepatitis have increased faecal numbers of E. faecalis. The presence of cytolysin-positive (cytolytic) E. faecalis correlated with the severity of liver disease and with mortality in patients with alcoholic hepatitis. Using humanized mice that were colonized with bacteria from the faeces of patients with alcoholic hepatitis, we investigated the therapeutic effects of bacteriophages that target cytolytic E. faecalis. We found that these bacteriophages decrease cytolysin in the liver and abolish ethanol-induced liver disease in humanized mice. Our findings link cytolytic E. faecalis with more severe clinical outcomes and increased mortality in patients with alcoholic hepatitis. We show that bacteriophages can specifically target cytolytic E. faecalis, which provides a method for precisely editing the intestinal microbiota. A clinical trial with a larger cohort is required to validate the relevance of our findings in humans, and to test whether this therapeutic approach is effective for patients with alcoholic hepatitis.


Assuntos
Bacteriófagos/fisiologia , Enterococcus faecalis/patogenicidade , Enterococcus faecalis/virologia , Microbioma Gastrointestinal , Hepatite Alcoólica/microbiologia , Hepatite Alcoólica/terapia , Terapia por Fagos , Alcoolismo/complicações , Alcoolismo/microbiologia , Animais , Enterococcus faecalis/isolamento & purificação , Etanol/efeitos adversos , Fígado Gorduroso/complicações , Fígado Gorduroso/microbiologia , Fezes/microbiologia , Feminino , Vida Livre de Germes , Hepatite Alcoólica/complicações , Hepatite Alcoólica/mortalidade , Hepatócitos/efeitos dos fármacos , Hepatócitos/patologia , Humanos , Fígado/efeitos dos fármacos , Fígado/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Perforina/metabolismo
18.
Microb Cell Fact ; 18(1): 188, 2019 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-31690323

RESUMO

BACKGROUND: Probiotic bacteria can provide health benefits when delivered in functional foods. This study involved isolation of lactic acid bacteria (LAB) from traditionally dried and salted anchovy fish and characterization of their survival in simulated gastrointestinal digestion. Promising strains were used to prepare fermented fish sausages which were then evaluated for cytotoxicity activity against two cancer cell-lines, antidiabetic activity as determined by α-amylase and α-glucosidase inhibition, and antioxidant and proteolytic activities in vitro, as compared to non-fermented control sausages. RESULTS: Out of 85 LAB obtained, 13 isolates with high tolerance to simulated gastrointestinal digestion were obtained, which were identified as Enterococcus spp. Four E. faecium strains, one E. faecalis, and one E. durans were used separately to make fermented fish sausages. The α-amylase and α-glucosidase inhibition from fish sausages fermented by Enterococcus spp. ranged from 29.2 to 68.7% and 23.9 to 41.4%, respectively, during 21 days of storage. The cytotoxicity activities against Caco2 and MCF-7 cells of fish sausages fermented with Enterococcus spp. ranged from 18.0 to 24% and 13.9 to 27.9%, respectively. Cytotoxicity activities correlated positively with proteolysis and antioxidant activities, α-amylase and α-glucosidase inhibition activities, but negatively with the pH in fermented fish sausages. Strains also exhibited antimicrobial activity against foodborne pathogens and presented no significant concerns with regards to antibiotic resistance or virulence gene content. CONCLUSIONS: Fish sausages fermented by potential probiotic isolates of Enterococcus spp. from dried fish had valuable health-promoting benefits compared with non-fermented control sausages.


Assuntos
Enterococcus faecalis , Enterococcus faecium , Enterococcus , Produtos Pesqueiros/microbiologia , Microbiologia de Alimentos , Probióticos , Linhagem Celular Tumoral , Diabetes Mellitus/terapia , Enterococcus/isolamento & purificação , Enterococcus/metabolismo , Enterococcus faecalis/isolamento & purificação , Enterococcus faecalis/metabolismo , Enterococcus faecium/isolamento & purificação , Enterococcus faecium/metabolismo , Humanos , Probióticos/isolamento & purificação , Probióticos/metabolismo
20.
Appl Environ Microbiol ; 85(22)2019 11 15.
Artigo em Inglês | MEDLINE | ID: mdl-31471308

RESUMO

Industrial farms are unique, human-created ecosystems that provide the perfect setting for the development and dissemination of antibiotic resistance. Agricultural antibiotic use amplifies naturally occurring resistance mechanisms from soil ecologies, promoting their spread and sharing with other bacteria, including those poised to become endemic within hospital environments. To better understand the role of enterococci in the movement of antibiotic resistance from farm to table to clinic, we characterized over 300 isolates of Enterococcus cultured from raw chicken meat purchased at U.S. supermarkets by the Consumers Union in 2013. Enterococcus faecalis and Enterococcus faecium were the predominant species found, and antimicrobial susceptibility testing uncovered striking levels of resistance to medically important antibiotic classes, particularly from classes approved by the FDA for use in animal production. While nearly all isolates were resistant to at least one drug, bacteria from meat labeled as raised without antibiotics had fewer resistances, particularly for E. faecium Whole-genome sequencing of 92 isolates revealed that both commensal- and clinical-isolate-like enterococcal strains were associated with chicken meat, including isolates bearing important resistance-conferring elements and virulence factors. The ability of enterococci to persist in the food system positions them as vehicles to move resistance genes from the industrial farm ecosystem into more human-proximal ecologies.IMPORTANCE Bacteria that contaminate food can serve as a conduit for moving drug resistance genes from farm to table to clinic. Our results show that chicken meat-associated isolates of Enterococcus are often multidrug resistant, closely related to pathogenic lineages, and harbor worrisome virulence factors. These drug-resistant agricultural isolates could thus represent important stepping stones in the evolution of enterococci into drug-resistant human pathogens. Although significant efforts have been made over the past few years to reduce the agricultural use of antibiotics, continued assessment of agricultural practices, including the roles of processing plants, shared breeding flocks, and probiotics as sources for resistance spread, is needed in order to slow the evolution of antibiotic resistance. Because antibiotic resistance is a global problem, global policies are needed to address this threat. Additional measures must be taken to mitigate the development and spread of antibiotic resistance elements from farms to clinics throughout the world.


Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana Múltipla/genética , Enterococcus/efeitos dos fármacos , Carne/microbiologia , Aves Domésticas/microbiologia , Agricultura , Animais , Galinhas/microbiologia , Ecossistema , Enterococcus/genética , Enterococcus/isolamento & purificação , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/genética , Enterococcus faecalis/isolamento & purificação , Enterococcus faecium/efeitos dos fármacos , Enterococcus faecium/genética , Enterococcus faecium/isolamento & purificação , Fazendas , Infecções por Bactérias Gram-Positivas/microbiologia , Infecções por Bactérias Gram-Positivas/transmissão , Instalações de Saúde , Humanos , Testes de Sensibilidade Microbiana , Alimentos Crus/microbiologia , Fatores de Virulência/genética
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