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1.
Clin Exp Dent Res ; 9(2): 341-348, 2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-36752174

RESUMO

OBJECTIVES: Successful endodontic therapy is based on the reduction of infecting bacteria by cleaning, shaping, and disinfecting of the root canal system, thus the use of intracanal dressing is necessary for optimal success of root canal treatment. This study was designed to evaluate the effect of chitosan and propolis as intracanal medicaments against Enterococcus faecalis compared to calcium hydroxide in primary root canals. MATERIAL AND METHODS: Ninety-six extracted primary second molars were collected. Teeth preparation was completed to size 30 K-file. They were randomly divided into four groups; (A): chitosan, (B): propolis, (C): calcium hydroxide, and (D): control group (saline). The tooth specimens were inoculated with E. faecalis. Then, tested materials were applied for all groups in accordance to the groups each tooth belonged to. Following this, the bacterial colonies were counted after 24 h, 72 h, and 1 week of applying dressing materials and incubation. Finally, one-way analysis of variance and Fisher's least significant difference tests were used for statistical comparisons between the groups at a significance level of .05. RESULTS: No statistically significant difference was found between groups A, B, and C for both 24 h and a week (p ≥ .05). Yet, a statistical difference between groups A, B, C, and D after 72 h and 1 week were seen (p ≤ .05). CONCLUSIONS: Chitosan and propolis medicaments were as effective as calcium hydroxide against E. faecalis in primary root canal treatment and might be considered as an alternative dressing material between treatment sessions.


Assuntos
Antibacterianos , Quitosana , Própole , Tratamento do Canal Radicular , Antibacterianos/farmacologia , Hidróxido de Cálcio/farmacologia , Quitosana/farmacologia , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/isolamento & purificação , Própole/farmacologia , Dente Decíduo/microbiologia , Tratamento do Canal Radicular/métodos , Humanos
2.
BMC Cancer ; 21(1): 1353, 2021 Dec 22.
Artigo em Inglês | MEDLINE | ID: mdl-34937552

RESUMO

BACKGROUND: The association between specific bacteria and colorectal cancer (CRC) has been proposed. Only a few studies have, however, investigated this relationship directly in colorectal tissue with conflicting results. So, we aimed to quantitate Streptococcus gallolyticus, Fusobacterium spp, Enterococcus faecalis and enterotoxigenic Bacteroides fragilis (ETBF) in formalin-fixed and paraffin-embedded (FFPE) colorectal tissue samples of Iranian CRC patients and healthy controls. METHODS: A total of 80 FFPE colorectal tissue samples of CRC patients (n = 40) and healthy controls (n = 40) were investigated for the presence and copy number of above bacterial species using quantitative PCR. Relative quantification was determined using ΔΔCT method and expressed as relative fold difference compared to reference gene. RESULTS: Relative abundance and copy number of E. faecalis and ETBF were significantly higher in CRC samples compared to control group. E. faecalis was more prevalent than ETBF in tumor samples. Frequency of ETBF and E. faecalis in late stages (III/IV) of cancer was significantly higher than early stages (I/II). We did not detect a significant difference in abundance of S. gallolyticus and Fusobacterium spp between two groups. CONCLUSION: Our study revealed the higher concentration of E. faecalis and ETBF in FFPE samples of CRC patients than controls. However, additional investigations on fecal and fresh colorectal cancer tissue samples are required to substantiate this correlation.


Assuntos
Infecções por Bacteroides/epidemiologia , Bacteroides fragilis/isolamento & purificação , Neoplasias Colorretais/microbiologia , Enterococcus faecalis/isolamento & purificação , Infecções por Bactérias Gram-Positivas/epidemiologia , Adulto , Idoso , Infecções por Bacteroides/diagnóstico , Infecções por Bacteroides/microbiologia , Infecções por Bacteroides/patologia , Bacteroides fragilis/genética , Estudos de Casos e Controles , Colo/microbiologia , Colo/patologia , Neoplasias Colorretais/diagnóstico , Neoplasias Colorretais/patologia , DNA Bacteriano/isolamento & purificação , Enterococcus faecalis/genética , Feminino , Infecções por Bactérias Gram-Positivas/diagnóstico , Infecções por Bactérias Gram-Positivas/microbiologia , Infecções por Bactérias Gram-Positivas/patologia , Humanos , Mucosa Intestinal/microbiologia , Mucosa Intestinal/patologia , Irã (Geográfico) , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Inclusão em Parafina , Prevalência
3.
Int J Mol Sci ; 22(21)2021 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-34768762

RESUMO

The use of phenicol antibiotics in animals has increased. In recent years, it has been reported that the transferable gene mediates phenicol-oxazolidinone resistance. This study analyzed the prevalence and characteristics of phenicol-oxazolidinone resistance genes in Enterococcus faecalis and Enterococcus faecium isolated from food-producing animals and meat in Korea in 2018. Furthermore, for the first time, we reported the genome sequence of E. faecalis strain, which possesses the phenicol-oxazolidinone resistance gene on both the chromosome and plasmid. Among the 327 isolates, optrA, poxtA, and fexA genes were found in 15 (4.6%), 8 (2.5%), and 17 isolates (5.2%), respectively. Twenty E. faecalis strains carrying resistance genes belonged to eight sequence types (STs), and transferability was found in 17 isolates. The genome sequences revealed that resistant genes were present in the chromosome or plasmid, or both. In strains EFS17 and EFS108, optrA was located downstream of the ermA and ant(9)-1 genes. The strains EFS36 and EFS108 harboring poxtA-encoding plasmid cocarried fexA and cfr(D). These islands also contained IS1216E or the transposon Tn554, enabling the horizontal transfer of the phenicol-oxazolidinone resistance with other antimicrobial-resistant genes. Our results suggest that it is necessary to promote the prudent use of antibiotics through continuous monitoring and reevaluation.


Assuntos
Anti-Infecciosos/farmacologia , Cloranfenicol/farmacologia , Farmacorresistência Bacteriana Múltipla/genética , Enterococcus faecalis/genética , Enterococcus faecium/genética , Carne/microbiologia , Oxazolidinonas/farmacologia , Animais , Bovinos/microbiologia , Biologia Computacional , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/isolamento & purificação , Enterococcus faecium/efeitos dos fármacos , Enterococcus faecium/isolamento & purificação , Análise de Alimentos , Transferência Genética Horizontal , Genes Bacterianos/efeitos dos fármacos , Genoma Bacteriano , Tipagem de Sequências Multilocus , Plasmídeos , República da Coreia , Suínos/microbiologia , Sequenciamento Completo do Genoma
4.
Microbiol Spectr ; 9(2): e0081721, 2021 10 31.
Artigo em Inglês | MEDLINE | ID: mdl-34668732

RESUMO

The use of enterococci as a fecal indicator bacterial group for public health risk assessment has been brought into question by recent studies showing that "naturalized" populations of Enterococcus faecalis exist in the extraenteric environment. The extent to which these naturalized E. faecalis organisms can confound water quality monitoring is unclear. To determine if strains isolated from different habitats display different survival strategies and responses, we compared the decay patterns of three E. faecalis isolates from the natural environment (environmental strains) against three human gut isolates (enteric strains) in laboratory mesocosms that simulate an oligotrophic, aerobic freshwater environment. Our results showed similar overall decay rates between enteric and environmental isolates based on viable plate and quantitative PCR (qPCR) counts. However, the enteric isolates exhibited a spike in copy number ratios of 16S rRNA gene transcripts to 16S rRNA gene DNA copies (rRNA:rDNA ratios) between days 1 and 3 of the mesocosm incubations that was not observed in environmental isolates, which could indicate a different stress response. Nevertheless, there was no strong evidence of differential gene expression between environmental and enteric isolates related to habitat adaptation in the accompanying mesocosm metatranscriptomes. Overall, our results provide novel information on how rRNA levels may vary over different growth conditions (e.g., standard lab versus oligotrophic) for this important indicator bacteria. We also observed some evidence for habitat adaptation in E. faecalis; however, this adaptation may not be substantial or consistent enough for integration in water quality monitoring. IMPORTANCE Enterococci are commonly used worldwide to monitor environmental fecal contamination and public health risk for waterborne diseases. However, closely related enterococci strains adapted to living in the extraenteric environment may represent a lower public health risk and confound water quality estimates. We developed an rRNA:rDNA viability assay for E. faecalis (a predominant species within this fecal group) and tested it against both enteric and environmental isolates in freshwater mesocosms to assess whether this approach can serve as a more sensitive water quality monitoring tool. We were unable to reliably distinguish the different isolate types using this assay under the conditions tested; thus, environmental strains should continue to be counted during routine water monitoring. However, this assay could be useful for distinguishing more recent (i.e., higher-risk) fecal pollution because rRNA levels significantly decreased after 1 week in all isolates.


Assuntos
Adaptação Fisiológica/fisiologia , DNA Ribossômico/genética , Enterococcus faecalis/genética , Microbioma Gastrointestinal/genética , RNA Ribossômico 16S/genética , Biologia Computacional/métodos , Enterococcus faecalis/isolamento & purificação , Monitoramento Ambiental , Fezes/microbiologia , Água Doce/microbiologia , Dosagem de Genes/genética , Humanos , Intestinos/microbiologia , Transcriptoma/genética , Microbiologia da Água , Qualidade da Água
5.
Microbiol Spectr ; 9(2): e0006721, 2021 10 31.
Artigo em Inglês | MEDLINE | ID: mdl-34585952

RESUMO

Detection of bacterial DNA within meconium is often cited as evidence supporting in utero colonization. However, many studies fail to adequately control for contamination. We aimed to define the microbial content of meconium under properly controlled conditions. DNA was extracted from 141 meconium samples and subjected to cpn60-based microbiome profiling, with controls to assess contamination throughout. Total bacterial loads of neonatal meconium, infant stool, and controls were compared by 16S rRNA quantitative PCR (qPCR). Viable bacteria within meconium were cultured, and isolate clonality was assessed by pulsed-field gel electrophoresis (PFGE). Meconium samples did not differ significantly from controls with respect to read numbers or taxonomic composition. Twenty (14%) outliers with markedly higher read numbers were collected significantly later after birth and appeared more like transitional stool than meconium. Total bacterial loads were significantly higher in stool than in meconium, which did not differ from that of sequencing controls, and correlated well with read numbers. Cultured isolates were most frequently identified as Staphylococcus epidermidis, Enterococcus faecalis, or Escherichia coli, with PFGE indicating high intraspecies diversity. Our findings highlight the importance of robust controls in studies of low microbial biomass samples and argue against meaningful bacterial colonization in utero. Given that meconium microbiome profiles could not be distinguished from sequencing controls, and that viable bacteria within meconium appeared uncommon and largely consistent with postnatal skin colonization, there does not appear to be a meconium microbiota. IMPORTANCE Much like the recent placental microbiome controversy, studies of neonatal meconium reporting bacterial communities within the fetal and neonatal gut imply that microbial colonization begins prior to birth. However, recent work has shown that placental microbiomes almost exclusively represent contamination from lab reagents and the environment. Here, we demonstrate that prior studies of neonatal meconium are impacted by the same issue, showing that the microbial content of meconium does not differ from negative controls that have never contained any biological material. Our culture findings similarly supported this notion and largely comprised bacteria normally associated with healthy skin. Overall, our work adds to the growing body of evidence against the in utero colonization hypothesis.


Assuntos
Bactérias/classificação , DNA Bacteriano/isolamento & purificação , Fezes/microbiologia , Mecônio/microbiologia , Microbiota/genética , Adulto , Bactérias/genética , Bactérias/isolamento & purificação , Carga Bacteriana , Biomassa , DNA Bacteriano/genética , Eletroforese em Gel de Campo Pulsado , Enterococcus faecalis/genética , Enterococcus faecalis/isolamento & purificação , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Feminino , Humanos , Recém-Nascido , Masculino , Gravidez , Pele/microbiologia , Staphylococcus epidermidis/genética , Staphylococcus epidermidis/isolamento & purificação
6.
ScientificWorldJournal ; 2021: 6664636, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34421400

RESUMO

Recently, the interest in donkey milk has increased considerably because it proved high nutritive and functional values of their ingredients. Its chemical composition is widely studied, but its microbiota, especially lactic acid bacteria, remains less studied. This study focuses on analyzing, isolating, and identifying lactic acid bacteria and evaluating their capacity to produce biomolecules with antibacterial activity. Among 44 strains identified, 43 are Gram-positive, and most are catalase-negative and cocci-shaped. Five strains were selected to evaluate their antibacterial activity against Listeria monocytogenes, Staphylococcus aureus, and Escherichia coli. Different induction methods allowed to amplify the antibacterial effects against these pathogenic strains.


Assuntos
Aerococcus/isolamento & purificação , Antibacterianos/farmacologia , Meios de Cultivo Condicionados/farmacologia , Enterococcus faecalis/isolamento & purificação , Enterococcus/isolamento & purificação , Leuconostoc mesenteroides/isolamento & purificação , Aerococcus/química , Aerococcus/metabolismo , Animais , Indústria de Laticínios/métodos , Enterococcus/química , Enterococcus/metabolismo , Enterococcus faecalis/química , Enterococcus faecalis/metabolismo , Equidae , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Escherichia coli/patogenicidade , Feminino , Microbiologia de Alimentos , Lactação/fisiologia , Leuconostoc mesenteroides/química , Leuconostoc mesenteroides/metabolismo , Listeria monocytogenes/efeitos dos fármacos , Listeria monocytogenes/crescimento & desenvolvimento , Listeria monocytogenes/patogenicidade , Testes de Sensibilidade Microbiana , Leite/microbiologia , Marrocos , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus aureus/patogenicidade
7.
Sci Rep ; 11(1): 16010, 2021 08 06.
Artigo em Inglês | MEDLINE | ID: mdl-34362987

RESUMO

Enterococcus faecalis is a life-threatening bacterium that resists high levels of antibiotics or chemical preservatives. In this study, we aimed to investigate the inactivation of E. faecalis in fresh pineapple juice (FPJ) with two different cold atmospheric plasmas (CAP) reinforced by H2O2/H2O cold vapor: a plasma jet and a surface dielectric barrier discharge (SDBD). CAP treatments for 300 s with plasma jet and 420 s with SDBD caused an 8.2 log reduction of E. faecalis. The concentration of reactive oxygen and nitrogen species induced in FPJ by plasmas was also evaluated. In terms of quality attributes of FPJ, no noticeable color changes (ΔE) were observed. Furthermore, a trifle of loss of organic content such as sugars and organic acids was observed after treatments. These results suggest that our rapid CAP strategy effectively inactivated E. faecalis in FPJ with no change of color and negligible effects on other physicochemical properties.


Assuntos
Ananas/efeitos dos fármacos , Antibacterianos/farmacologia , Enterococcus faecalis/efeitos dos fármacos , Sucos de Frutas e Vegetais/análise , Gases em Plasma/farmacologia , Ananas/microbiologia , Enterococcus faecalis/isolamento & purificação , Sucos de Frutas e Vegetais/microbiologia
8.
Ann Clin Microbiol Antimicrob ; 20(1): 49, 2021 Jul 28.
Artigo em Inglês | MEDLINE | ID: mdl-34321002

RESUMO

Clustered regularly interspaced short palindromic repeat (CRISPR)-Cas systems are one of the factors which can contribute to limiting the development and evolution of antibiotic resistance in bacteria. There are three genomic loci of CRISPR-Cas in Enterococcus faecalis. In this study, we aimed to assess correlation of the CRISPR-Cas system distribution with the acquisition of antibiotic resistance among E. faecalis isolates. A total of 151 isolates of E. faecalis were collected from urinary tract infections (UTI) and dental-root canal (DRC). All isolates were screened for phenotypic antibiotic resistance. In addition, antibiotic resistance genes and CRISPR loci were screened by using polymerase chain reaction. Genomic background of the isolates was identified by random amplified polymorphic DNA (RAPD)-PCR. The number of multidrug-resistant E. faecalis strains were higher in UTI isolates than in DRC isolates. RAPD-PCR confirmed that genomic background was diverse in UTI and DRC isolates used in this study. CRISPR loci were highly accumulated in gentamycin-, teicoplanin-, erythromycin-, and tetracycline-susceptible strains. In concordance with drug susceptibility, smaller number of CRISPR loci were identified in vanA, tetM, ermB, aac6'-aph(2"), aadE, and ant(6) positive strains. These data indicate a negative correlation between CRISPR-cas loci and antibiotic resistance, as well as, carriage of antibiotic resistant genes in both of UTI and DRC isolates.


Assuntos
Antibacterianos/farmacologia , Sistemas CRISPR-Cas , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas , Farmacorresistência Bacteriana Múltipla/genética , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/genética , Proteínas de Bactérias/genética , Enterococcus faecalis/isolamento & purificação , Genótipo , Gentamicinas , Humanos , Infecções Urinárias
9.
BMC Infect Dis ; 21(1): 531, 2021 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-34090359

RESUMO

BACKGROUND: Group B Streptococcus (GBS) is an important cause of invasive infection in neonates and infants. Cerebrospinal fluid (CSF) findings and culture may not show evidence of infection early in GBS meningitis. Next-generation sequencing (NGS) has the potential to detect microbial genetic material in patients with infectious diseases. We report two cases of infantile sepsis of GBS meningitis with negative results for CSF culture tests, but positive results for NGS analysis. CASE PRESENTATION: Patient 1 was a 22-day-old male infant diagnosed with sepsis and meningitis. His CSF findings showed pleocytosis, decreased glucose, and increased protein levels. However, CSF and blood culture results at admission were negative. He received a total of 3 weeks of treatment with ampicillin and cefotaxime, and showed clinical improvement. GBS was detected through NGS analysis of CSF collected at admission. Patient 2 was a 51-day-old male infant with sepsis. CSF findings on admission were normal, and blood and CSF cultures were also negative. Intravenous ampicillin and cefotaxime treatment were initiated. Treatment was de-escalated to ampicillin alone because Enterococcus faecalis was cultured from urine. He was discharged after a total of 1 week of antibiotic treatment. Six days after discharge, he was re-hospitalized for sepsis. Blood and CSF cultures were negative, and E. faecalis was again cultured from urine. He received a total of 3 weeks of ampicillin treatment for enterococcal-induced nephritis and did not relapse thereafter. NGS pathogen searches were retrospectively performed on both blood and CSF collected at the first and second admission. GBS was detected in the CSF collected at the first admission, but no significant pathogen was detected in the other samples. Inadequate treatment for GBS meningitis at the first admission may have caused the recurrence of the disease. CONCLUSION: Infantile sepsis may present bacterial meningitis that is not diagnosed by either culture testing or CSF findings. NGS analysis for CSF may be useful for confirming the diagnosis of bacterial meningitis.


Assuntos
Antibacterianos/uso terapêutico , Meningites Bacterianas/diagnóstico , Meningites Bacterianas/tratamento farmacológico , Infecções Estreptocócicas/diagnóstico , Infecções Estreptocócicas/tratamento farmacológico , Ampicilina/uso terapêutico , Cefotaxima/uso terapêutico , Líquido Cefalorraquidiano/microbiologia , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/isolamento & purificação , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Lactente , Recém-Nascido , Masculino , Meningites Bacterianas/microbiologia , Estudos Retrospectivos , Sepse/diagnóstico , Sepse/tratamento farmacológico , Sepse/microbiologia , Streptococcus agalactiae/efeitos dos fármacos , Streptococcus agalactiae/genética , Streptococcus agalactiae/isolamento & purificação , Urina/microbiologia
10.
Diagn Microbiol Infect Dis ; 101(1): 115433, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34139401

RESUMO

Enterococcus faecalis can cause infective endocarditis and other complicated infections. We prospectively investigate the incidence of persistent bacteremia with E. faecalis. Of 50 episodes with monomicrobial E. faecalis bacteremia the control blood culture after 48 to 72 hours was positive in 5 episodes (10%) of which 4 had a complicated focal infection.


Assuntos
Bacteriemia/diagnóstico , Hemocultura , Enterococcus faecalis/isolamento & purificação , Infecções por Bactérias Gram-Positivas/diagnóstico , Idoso , Idoso de 80 Anos ou mais , Bacteriemia/epidemiologia , Bacteriemia/microbiologia , Endocardite Bacteriana/diagnóstico , Endocardite Bacteriana/epidemiologia , Endocardite Bacteriana/microbiologia , Feminino , Infecção Focal/diagnóstico , Infecção Focal/epidemiologia , Infecção Focal/microbiologia , Infecções por Bactérias Gram-Positivas/epidemiologia , Infecções por Bactérias Gram-Positivas/microbiologia , Humanos , Incidência , Masculino , Estudos Prospectivos
11.
Chest ; 159(6): e377-e380, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-34099153

RESUMO

CASE PRESENTATION: An 80-year old man presented to the ED after being found down in his home. He had gone to the restroom to perform straight catheterization, as he normally does for his enlarged prostate, and was found slumped over and confused. On arrival to the ED, he was found to be in atrial fibrillation with rapid ventricular response, febrile, and hypotensive. CT imaging of his head, chest, and abdomen-pelvis was obtained. He was started on broad-spectrum antibiotics and norepinephrine infusion for presumed urinary septic shock and admitted to the medical critical care unit for further management.


Assuntos
Anti-Infecciosos/administração & dosagem , Valva Aórtica , Endocardite , Enterococcus faecalis/isolamento & purificação , Próteses Valvulares Cardíacas/microbiologia , Infecções Relacionadas à Prótese , Choque Séptico , Idoso de 80 Anos ou mais , Valva Aórtica/diagnóstico por imagem , Valva Aórtica/fisiopatologia , Valva Aórtica/cirurgia , Fibrilação Atrial/diagnóstico , Fibrilação Atrial/fisiopatologia , Confusão/diagnóstico , Confusão/etiologia , Diagnóstico Diferencial , Ecocardiografia/métodos , Endocardite/microbiologia , Endocardite/fisiopatologia , Endocardite/terapia , Humanos , Masculino , Tomografia por Emissão de Pósitrons/métodos , Infecções Relacionadas à Prótese/microbiologia , Infecções Relacionadas à Prótese/fisiopatologia , Infecções Relacionadas à Prótese/terapia , Choque Séptico/microbiologia , Choque Séptico/fisiopatologia , Choque Séptico/terapia , Resultado do Tratamento , Sinais Vitais
12.
Exp Eye Res ; 208: 108614, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-33971221

RESUMO

The aim of this study was to investigate the efficacy of intravitreal povidone-iodine (PI) in the treatment of vancomycin-resistant Enterococcus faecalis (VRE) endophthalmitis. Fifty New Zealand white rabbits were divided into 5 groups (n = 10 in each group). After the induction of endophthalmitis using VRE (minimum inhibitory concentration [MIC] ≥ 40 µg/mL) in the right eye, Group A, B, C, and D received intravitreal injections of 0.1% PI, 0.3% PI, 0.05% vancomycin, and 0.5% vancomycin, respectively. Eyes in Group E were used as controls. Fundus photography, vitreous culture, electroretinography (ERG), and histologic examinations of the retina were conducted on day 14. A marked improvement in endophthalmitis was observed in Group A, B, C and D, compared to Group E. Fundus photographs showed mild vitreous opacities in Group A and B, and moderate vitreous opacity in Group C. All eyes in Group D had a clear vitreous. In vitreous culture, bacterial growth was found in 6 eyes (100, 200, 200, 400, 500, and 500 colony-forming units) in Group C, but not in Groups A, B, or D. ERG and histological examination also indicated intraocular damage in Group C. Our results show that intravitreal injection of PI, even at low concentrations, was effective for treatment of VRE endophthalmitis, although some vitreous opacity remained. Intravitreal vancomycin injection was also useful to treat resistant strains, if used at a higher concentration within the safety threshold.


Assuntos
Endoftalmite/tratamento farmacológico , Enterococcus faecalis/isolamento & purificação , Infecções Oculares Bacterianas/tratamento farmacológico , Infecções por Bactérias Gram-Positivas/tratamento farmacológico , Povidona-Iodo/administração & dosagem , Resistência a Vancomicina , Animais , Anti-Infecciosos Locais/administração & dosagem , Contagem de Colônia Microbiana , Modelos Animais de Doenças , Eletrorretinografia , Endoftalmite/microbiologia , Endoftalmite/patologia , Infecções Oculares Bacterianas/microbiologia , Infecções Oculares Bacterianas/patologia , Infecções por Bactérias Gram-Positivas/microbiologia , Infecções por Bactérias Gram-Positivas/patologia , Injeções Intravítreas , Coelhos , Retina/patologia , Corpo Vítreo/microbiologia , Corpo Vítreo/patologia
13.
Elife ; 102021 05 25.
Artigo em Inglês | MEDLINE | ID: mdl-34032213

RESUMO

Upon exposure to harmful microorganisms, hosts engage in protective molecular and behavioral immune responses, both of which are ultimately regulated by the nervous system. Using the nematode Caenorhabditis elegans, we show that ingestion of Enterococcus faecalis leads to a fast pathogen avoidance behavior that results in aversive learning. We have identified multiple sensory mechanisms involved in the regulation of avoidance of E. faecalis. The G-protein coupled receptor NPR-1-dependent oxygen-sensing pathway opposes this avoidance behavior, while an ASE neuron-dependent pathway and an AWB and AWC neuron-dependent pathway are directly required for avoidance. Colonization of the anterior part of the intestine by E. faecalis leads to AWB and AWC mediated olfactory aversive learning. Finally, two transient receptor potential melastatin (TRPM) channels, GON-2 and GTL-2, mediate this newly described rapid pathogen avoidance. These results suggest a mechanism by which TRPM channels may sense the intestinal distension caused by bacterial colonization to elicit pathogen avoidance and aversive learning by detecting changes in host physiology.


Assuntos
Aprendizagem da Esquiva , Caenorhabditis elegans/microbiologia , Enterococcus faecalis/patogenicidade , Intestinos/microbiologia , Canais de Cátion TRPM/fisiologia , Animais , Enterococcus faecalis/isolamento & purificação , Interações Hospedeiro-Patógeno , Intestinos/patologia , Neurônios/metabolismo , Virulência
15.
Int J Mol Sci ; 22(9)2021 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-33946710

RESUMO

Considering the advent of antibiotic resistance, the study of bacterial metabolic behavior stimulated by novel antimicrobial agents becomes a relevant tool to elucidate involved adaptive pathways. Profiling of volatile metabolites was performed to monitor alterations of bacterial metabolism induced by biosynthesized silver nanoparticles (bio-AgNPs). Escherichia coli, Enterococcus faecalis, Klebsiella pneumoniae and Proteus mirabilis were isolated from pressure ulcers, and their cultures were prepared in the presence/absence of bio-AgNPs at 12.5, 25 and 50 µg mL-1. Headspace solid phase microextraction associated to gas chromatography-mass spectrometry was the employed analytical platform. At the lower concentration level, the agent promoted positive modulation of products of fermentation routes and bioactive volatiles, indicating an attempt of bacteria to adapt to an ongoing suppression of cellular respiration. Augmented response of aldehydes and other possible products of lipid oxidative cleavage was noticed for increasing levels of bio-AgNPs. The greatest concentration of agent caused a reduction of 44 to 80% in the variety of compounds found in the control samples. Pathway analysis indicated overall inhibition of amino acids and fatty acids routes. The present assessment may provide a deeper understanding of molecular mechanisms of bio-AgNPs and how the metabolic response of bacteria is untangled.


Assuntos
Bactérias/efeitos dos fármacos , Bactérias/metabolismo , Nanopartículas Metálicas/uso terapêutico , Lesão por Pressão/tratamento farmacológico , Lesão por Pressão/microbiologia , Prata/uso terapêutico , Compostos Orgânicos Voláteis/metabolismo , Antibacterianos/uso terapêutico , Bactérias/isolamento & purificação , Farmacorresistência Bacteriana , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/isolamento & purificação , Enterococcus faecalis/metabolismo , Escherichia coli/efeitos dos fármacos , Escherichia coli/isolamento & purificação , Escherichia coli/metabolismo , Humanos , Técnicas In Vitro , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/isolamento & purificação , Klebsiella pneumoniae/metabolismo , Redes e Vias Metabólicas/efeitos dos fármacos , Metaboloma/efeitos dos fármacos , Metabolômica , Testes de Sensibilidade Microbiana , Proteus mirabilis/efeitos dos fármacos , Proteus mirabilis/isolamento & purificação , Proteus mirabilis/metabolismo , Compostos Orgânicos Voláteis/química , Compostos Orgânicos Voláteis/classificação
16.
Toxins (Basel) ; 13(3)2021 03 02.
Artigo em Inglês | MEDLINE | ID: mdl-33801318

RESUMO

Wound necrosis and secondary infection are common complications after Naja atra bites. Clinical tools to evaluate the infection risk after Taiwan cobra bites are lacking. In this Cobra BITE study, we investigated the prevalence of wound infection, bacteriology, and corresponding antibiotic usage in patients presenting with Taiwan cobra snakebites. Patients with wound infection lacking tissue necrosis were included in developing Cobra BITE score utilizing univariate and multiple logistic regression, as patients with wound necrosis require antibiotics for infection treatment. 8,295,497 emergency department visits occurred in the span of this study, with 195 of those patients being diagnosed as having cobra bites. Of these patients, 23 had wound necrosis, and 30 had wound infection, resulting in a wound infection rate of 27.2% (53/195). Enterococcus faecalis and Morganella morganii were the main bacteria identified in the culture report regardless of whether patients' wounds had necrosis. As per our Cobra BITE score, the three factors predicting secondary wound infection after cobra bites are hospital admission, a white blood cell count (in 103/µL) × by neu-trophil-lymphocyte ratio value of ≥114.23, and the use of antivenin medication. The area under the receiver operating characteristic curve for the Cobra BITE score system was 0.88; ideal sensitivity and specificity were 0.89 and 0.76. This scoring system enables the assessment of wound infections after N. atra bites, and it could be modified and improved in the future for other Naja spp. bites.


Assuntos
Antibacterianos/uso terapêutico , Antivenenos/uso terapêutico , Venenos Elapídicos/antagonistas & inibidores , Enterococcus faecalis/efeitos dos fármacos , Morganella morganii/efeitos dos fármacos , Naja naja , Mordeduras de Serpentes/tratamento farmacológico , Infecção dos Ferimentos/tratamento farmacológico , Adulto , Idoso , Animais , Técnicas de Apoio para a Decisão , Venenos Elapídicos/imunologia , Enterococcus faecalis/isolamento & purificação , Feminino , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Morganella morganii/isolamento & purificação , Necrose , Estudos Retrospectivos , Medição de Risco , Fatores de Risco , Mordeduras de Serpentes/diagnóstico , Mordeduras de Serpentes/microbiologia , Resultado do Tratamento , Infecção dos Ferimentos/diagnóstico , Infecção dos Ferimentos/microbiologia
17.
Sci Rep ; 11(1): 8802, 2021 04 22.
Artigo em Inglês | MEDLINE | ID: mdl-33888775

RESUMO

Urinary tract infection is one of the most common bacterial infections leading to increased morbidity, mortality and societal costs. Current diagnostics exacerbate this problem due to an inability to provide timely pathogen identification. Surface enhanced Raman spectroscopy (SERS) has the potential to overcome these issues by providing immediate bacterial classification. To date, achieving accurate classification has required technically complicated processes to capture pathogens, which has precluded the integration of SERS into rapid diagnostics. This work demonstrates that gold-coated membrane filters capture and aggregate bacteria, separating them from urine, while also providing Raman signal enhancement. An optimal gold coating thickness of 50 nm was demonstrated, and the diagnostic performance of the SERS-active filters was assessed using phantom urine infection samples at clinically relevant concentrations (105 CFU/ml). Infected and uninfected (control) samples were identified with an accuracy of 91.1%. Amongst infected samples only, classification of three bacteria (Escherichia coli, Enterococcus faecalis, Klebsiella pneumoniae) was achieved at a rate of 91.6%.


Assuntos
Análise Espectral Raman/métodos , Infecções Urinárias/tratamento farmacológico , Enterococcus faecalis/isolamento & purificação , Escherichia coli/isolamento & purificação , Humanos , Klebsiella pneumoniae/isolamento & purificação , Propriedades de Superfície , Infecções Urinárias/microbiologia
18.
Emerg Microbes Infect ; 10(1): 929-938, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33913790

RESUMO

Context: Today, infective endocarditis (IE) caused by Enterococcus faecalis represents 10% of all IE and is marked by its difficult management and the frequency of relapses. Although the precise reasons for that remain to be elucidated, the evolution of the culprit strain under selective pressure through microdiversification could be, at least in part, involved. Material and methods: To further study the in situ genetic microdiversity and its possible phenotypic manifestations in E. faecalis IE, we sequenced and compared multiple isolates from the valves, blood culture and joint fluid of five patients who underwent valvular surgery. Growth rate and early biofilm production of selected isolates were also compared. Results: By sequencing a total of 58 E. faecalis genomes, we detected a considerable genomic microdiversity, not only among strains from different anatomical origins, but also between isolates from the same studied cardiac valves. Interestingly, deletions of thousands of bases including the well-known virulence factors ebpA/B/C, and srtC, as well as other large prophage sequences containing genes coding for proteins implicated in platelet binding (PlbA and PlbB) were evidenced. The study of mutations helped unveil common patterns in genes related to the cell cycle as well as central metabolism, suggesting an evolutionary convergence in these isolates. As expected, such modifications were associated with a significant impact on the in-vitro phenotypic heterogeneity, growth, and early biofilm production. Conclusion: Genome modifications associated with phenotypic variations may allow bacterial adaptation to both antibiotic and immune selective pressures, and thus promote relapses.


Assuntos
Endocardite Bacteriana/microbiologia , Enterococcus faecalis/classificação , Variação Genética , Infecções por Bactérias Gram-Positivas/microbiologia , Idoso , Idoso de 80 Anos ou mais , Códon sem Sentido , Enterococcus faecalis/genética , Enterococcus faecalis/isolamento & purificação , Feminino , Genoma Bacteriano , Valvas Cardíacas/microbiologia , Humanos , Masculino , Fenótipo , Deleção de Sequência , Sequenciamento Completo do Genoma
19.
Eur J Med Chem ; 219: 113402, 2021 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-33845234

RESUMO

A collection of potent antimicrobials consisting of novel 1,3-bis-benzoic acid and trifluoromethyl phenyl derived pyrazoles has been synthesized and tested for antibacterial activity. The majority of trifluoromethyl phenyl derivatives are highly potent growth inhibitors of Gram-positive bacteria and showed low toxicity to human cultured cells. In particular, two compounds (59 and 74) were selected for additional studies. These compounds are highly effective against Staphylococcus aureus as shown by a low minimum inhibitory concentration (MIC), a bactericidal effect in time-kill assays, moderate inhibition of biofilm formation as well as biofilm destruction, and a bactericidal effect against stationary phase cells representing non-growing persister cells. Multistep resistance assays showed a very low tendency for S. aureus and Enterococcus faecalis to develop resistance through mutation. Additionally, in vivo mouse model studies showed no harmful effects at doses up to 50 mg/kg using 14 blood plasma organ toxicity markers or TUNEL assay in liver and kidney. Investigations into the mode of action by performing macromolecular synthesis inhibition studies showed a broad range of inhibitory effects, suggesting targets that have a global effect on bacterial cell function.


Assuntos
Compostos de Anilina/química , Antibacterianos/farmacologia , Bactérias Gram-Positivas/efeitos dos fármacos , Pirazóis/química , Compostos de Anilina/síntese química , Compostos de Anilina/farmacologia , Antibacterianos/química , Biofilmes/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Farmacorresistência Bacteriana/efeitos dos fármacos , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/isolamento & purificação , Enterococcus faecalis/fisiologia , Bactérias Gram-Positivas/isolamento & purificação , Humanos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Testes de Sensibilidade Microbiana , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/isolamento & purificação , Staphylococcus aureus/fisiologia , Relação Estrutura-Atividade
20.
PLoS One ; 16(4): e0250084, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33852628

RESUMO

Enterococcus faecalis are hospital-associated opportunistic pathogens and also causative agents of post-operative endophthalmitis. Patients with enterococcal endophthalmitis often have poor visual outcomes, despite appropriate antibiotic therapy. Here we investigated the genomic and phenotypic characteristics of E. faecalis isolates collected from 13 patients treated at the University of Pittsburgh Medical Center Eye Center over 19 years. Comparative genomic analysis indicated that patients were infected with E. faecalis belonging to diverse multi-locus sequence types (STs) and resembled E. faecalis sampled from clinical, commensal, and environmental sources. We identified known E. faecalis virulence factors and antibiotic resistance genes in each genome, including genes conferring resistance to aminoglycosides, erythromycin, and tetracyclines. We assessed all isolates for their cytolysin production, biofilm formation, and antibiotic susceptibility, and observed phenotypic differences between isolates. Fluoroquinolone and cephalosporin susceptibilities were particularly variable between isolates, as were biofilm formation and cytolysin production. In addition, we found evidence of E. faecalis adaptation during recurrent endophthalmitis by identifying genetic variants that arose in sequential isolates sampled over eight months from the same patient. We identified a mutation in the DNA mismatch repair gene mutS that was associated with an increased rate of spontaneous mutation in the final isolate from the patient. Overall this study documents the genomic and phenotypic variability among E. faecalis causing endophthalmitis, as well as possible adaptive mechanisms underlying bacterial persistence during recurrent ocular infection.


Assuntos
Endoftalmite/microbiologia , Enterococcus faecalis/genética , Enterococcus faecalis/metabolismo , Idoso de 80 Anos ou mais , Antibacterianos/farmacologia , Farmacorresistência Bacteriana/genética , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Endoftalmite/metabolismo , Enterococcus/genética , Enterococcus faecalis/isolamento & purificação , Feminino , Genômica/métodos , Infecções por Bactérias Gram-Positivas/microbiologia , Humanos , Masculino , Testes de Sensibilidade Microbiana , Fatores de Virulência/genética
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