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1.
Food Microbiol ; 86: 103335, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31703853

RESUMO

The presence of eight common structural enterocin genes, singly or in varying combinations, in the genome of 15 antagonistic Enterococcus spp. previously isolated from artisan Greek Graviera and Galotyri retail cheeses was tested and associated with the mode of enterocin (Ent+) antilisterial activity of each isolate in three liquid culture media. The isolates were assigned to nine distinct strain genotypes of E. faecium (4 strains), E. durans (2) and E. faecalis (3). All strains were non-hemolytic, except for a cyl-positive E. faecalis genotype isolated from Galotyri cheese, which was strongly listericidal. All other strains varied from being listeriostatic to weakly listericidal in MRS and M17 broth, whereas all failed to inhibit listerial growth in skim milk. Two E. faecium strains retained strong Ent+ activity following neutralization and filter-sterilization of their MRS or M17 co-culture supernatants, whereas, all others required contact or proximity of their viable cells with L. monocytogenes cells in order to display activity. Additional studies to evaluate safety and potential synergistic effects of each strain genotype with starter LAB species in real milk environments will reveal the most active and truly harmless Enterococcus genotypes to be applied as co-starter or bioprotective adjunct cultures in traditional Greek cheese technologies.


Assuntos
Queijo/microbiologia , Enterococcus/química , Listeria monocytogenes/efeitos dos fármacos , Leite/microbiologia , Animais , Hidrocarbonetos Aromáticos com Pontes/química , Hidrocarbonetos Aromáticos com Pontes/metabolismo , Hidrocarbonetos Aromáticos com Pontes/farmacologia , Bovinos , Meios de Cultura/química , Meios de Cultura/metabolismo , Enterococcus/genética , Enterococcus/isolamento & purificação , Enterococcus/metabolismo , Grécia , Listeria monocytogenes/crescimento & desenvolvimento
2.
BMC Infect Dis ; 19(1): 1062, 2019 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-31852453

RESUMO

BACKGROUND: To investigate the cumulative incidence of and factors associated with mortality among patients with infective endocarditis (IE) at Thailand's largest national tertiary referral center. METHODS: Medical charts of adult patients diagnosed with IE by Duke criteria at Siriraj Hospital during January 2005 to May 2015 were retrospectively reviewed. RESULTS: Of 380 patients, 66.3% had definite IE, and 81.3% had native valve IE (NVE). Cumulative IE incidence was 5.67/1000 admissions. The most common pathogens were viridans group streptococci (VGS) (39.7%), methicillin-sensitive Staphylococcus aureus (MSSA) (13.1%), and beta-hemolytic streptococci (11.5%) in NVE; and, MSSA (20.3%), VGS (20.3%), and Enterococcus spp. (16.9%) in prosthetic valve (PVE) or device-related IE (DRIE). Overall in-hospital mortality was 18.4%. Mortality was significantly higher in PVE/DRIE than in NVE (26.8% vs. 16.5%, p = 0.047). End-stage renal disease (ESRD) (aOR: 9.43, 95% CI: 2.36-37.70), diabetes mellitus (DM) (aOR: 2.81, 95% CI: 1.06-7.49), neurological complication (aOR: 14.16, 95% CI: 5.11-39.22), congestive heart failure (aOR: 4.32, 95% CI: 1.91-9.75), hospital-acquired infection (aOR: 3.78, 95% CI: 1.66-8.57), renal complication (aOR: 3.12, 95%CI: 1.32-7.37), and other complication during admission (aOR: 3.28, 95% CI: 1.41-7.61) were independently associated with mortality. CONCLUSIONS: The incidence of IE, and the mortality rate among those diagnosed with IE are both increasing in Thailand - particularly among those with PVE or DRIE. End-stage renal disease, diabetes mellitus, and development of IE-related complications during admission were found to be independent predictors of mortality.


Assuntos
Endocardite Bacteriana/epidemiologia , Enterococcus/isolamento & purificação , Infecções Estafilocócicas/epidemiologia , Staphylococcus aureus/isolamento & purificação , Infecções Estreptocócicas/epidemiologia , Streptococcus/isolamento & purificação , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Infecção Hospitalar , Endocardite Bacteriana/microbiologia , Endocardite Bacteriana/mortalidade , Feminino , Mortalidade Hospitalar , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/mortalidade , Infecções Estreptocócicas/microbiologia , Infecções Estreptocócicas/mortalidade , Tailândia/epidemiologia , Estreptococos Viridans/isolamento & purificação , Adulto Jovem
3.
BMC Infect Dis ; 19(1): 1083, 2019 Dec 27.
Artigo em Inglês | MEDLINE | ID: mdl-31881851

RESUMO

BACKGROUND: Treatment of enterococcal periprosthetic joint infections (PJI) is challenging due to non-standardized management strategies and lack of biofilm-active antibiotics. The optimal surgical and antimicrobial therapy are unknown. Therefore, we evaluated characteristics and outcome of enterococcal PJI. METHODS: Consecutive patients with enterococcal PJI from two specialized orthopedic institutions were retrospectively analyzed. Both institutions are following the same diagnostic and treatment concepts. The probability of relapse-free survival was estimated using Kaplan-Meier survival curves and compared by log-rank test. Treatment success was defined by absence of relapse or persistence of PJI due to enterococci or death related to enterococcal PJI. Clinical success was defined by the infection-free status, no subsequent surgical intervention for persistent or perioperative infection after re-implantation and no PJI-related death within 3 months. RESULTS: Included were 75 enterococcal PJI episodes, involving 41 hip, 30 knee, 2 elbow and 2 shoulder prostheses. PJI occurred postoperatively in 61 episodes (81%), hematogenously in 13 (17%) and by contiguous spread in one. E. faecalis grew in 64 episodes, E. faecium in 10 and E. casseliflavus in one episode(s). Additional microorganism(s) were isolated in 38 patients (51%). Enterococci were susceptible to vancomycin in 73 of 75 isolates (97%), to daptomycin in all 75 isolates, and to fosfomycin in 21 of 22 isolates (96%). The outcome data was available for 66 patients (88%). The treatment success after 3 years was 83.7% (95% confidence interval [CI]; 76.1-96.7%) and the clinical success was 67.5% (95% CI; 57.3-80.8%). In 11 patients (17%), a new PJI episode caused by a different pathogen occurred. All failures occurred within 3 years after surgery. CONCLUSION: About half of enterococcal PJI were polymicrobial infections. The treatment success was high (84%). All treatment failures occurred within the first 3 years after revision surgery. Interestingly, 17% of patients experienced a new PJI caused by another pathogen at a later stage. TRIAL REGISTRATION: The study was retrospectively registered with the public clinical trial identification NCT0253022 at https://www.clinicaltrials.gov on 15 July 2015.


Assuntos
Artroplastia de Substituição/efeitos adversos , Infecções Bacterianas/microbiologia , Coinfecção/microbiologia , Enterococcus/isolamento & purificação , Infecções Relacionadas à Prótese/microbiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/uso terapêutico , Infecções Bacterianas/tratamento farmacológico , Biofilmes/efeitos dos fármacos , Coinfecção/tratamento farmacológico , Intervalo Livre de Doença , Feminino , Seguimentos , Fosfomicina/uso terapêutico , Humanos , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Infecções Relacionadas à Prótese/tratamento farmacológico , Recidiva , Reoperação/efeitos adversos , Estudos Retrospectivos , Falha de Tratamento
4.
BMC Infect Dis ; 19(1): 1066, 2019 Dec 19.
Artigo em Inglês | MEDLINE | ID: mdl-31856747

RESUMO

BACKGROUND: Symptomatic and asymptomatic enteric infections in early childhood are associated with negative effects on childhood growth and development, especially in low and middle-income countries, and food may be an important transmission route. Although basic food hygiene practices might reduce exposure to faecal pathogens and resulting infections, there have been few rigorous interventions studies to assess this, and no studies in low income urban settings where risks are plausibly very high. The aim of this study is to evaluate the impact of a novel infant food hygiene intervention on infant enteric infections and diarrhoea in peri-urban settlements of Kisumu, Kenya. METHODS: This is a cluster randomized control trial with 50 clusters, representing the catchment areas of Community Health Volunteers (CHVs), randomly assigned to intervention or control, and a total of 750 infants recruited on a rolling basis at 22 weeks of age and then followed for 15 weeks. The intervention targeted four key caregiver behaviours related to food hygiene: 1) hand washing with soap before infant food preparation and feeding; 2) bringing all infant food to the boil before feeding, including when reheating or reserving; 3) storing all infant food in sealed containers; and, 4) using only specific utensils for infant feeding which are kept separate and clean. RESULTS: The primary outcome of interest is the prevalence of one or more of 23 pre-specified enteric infections, determined using quantitative real-time polymerase chain reaction for enteric pathogen gene targets. In addition, infant food samples were collected at 33 weeks, and faecal indicator bacteria (Enterococcus) isolated and enumerated to assess the impact of the intervention on infant food contamination. CONCLUSION: To our knowledge this is the first randomized controlled trial to assess the effect of an infant food hygiene intervention on enteric infections in a high burden, low income urban setting. Our trial responds to growing evidence that food may be a key pathway for early childhood enteric infection and disease and that basic food hygiene behaviours may be able to mitigate these risks. The Safe Start trial seeks to provide new evidence as to whether a locally appropriate infant food hygiene intervention delivered through the local health extension system can improve the health of young children. TRIAL REGISTRATION: The trial was registered at clinicaltrial.gov on March 16th 2018 before enrolment of any participants (https://clinicaltrials.gov/ct2/show/NCT03468114).


Assuntos
Diarreia/epidemiologia , Diarreia/microbiologia , Enterite/epidemiologia , Enterite/microbiologia , Desinfecção das Mãos/métodos , Pobreza , Cuidadores , Culinária , Diarreia/prevenção & controle , Enterite/prevenção & controle , Enterococcus/isolamento & purificação , Fezes/microbiologia , Feminino , Contaminação de Alimentos/prevenção & controle , Armazenamento de Alimentos , Doenças Transmitidas por Alimentos/microbiologia , Doenças Transmitidas por Alimentos/prevenção & controle , Humanos , Lactente , Controle de Infecções , Quênia/epidemiologia , Masculino , Saúde Pública , Sabões , Saúde da População Urbana
5.
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
6.
Int J Mol Sci ; 20(17)2019 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-31480694

RESUMO

A comparative study on potential risks was carried out in a collection of 50 enterococci isolated from faeces of mice fed a standard or a high-fat diet enriched with extra virgin olive oil, refined olive oil or butter, at the beginning, after six weeks and after twelve weeks of experiments. Strains were biochemically assessed and genetically characterized. E. faecalis and E. casseliflavus were the most frequently isolated species in any diet and time points. Apart from the fact of not having isolated any strain from the virgin olive oil group during the last balance, we found statistically significant differences p < 0 . 05 among the diets in the percentage of antibiotic resistance and in the presence of the enterococcal surface protein gene (esp), as well as a tendency p < 0 . 1 for the presence of the tyrosine decarboxylase gene (tdc) to increase over time in the group of isolates from the standard diet. When the resistance of the strains to virgin or refined olive oil was studied, only the group of enterococci from high fat diets showed a significantly higher percentage of resistance to refined olive oil p < 0 . 05 , while both types of oil equally inhibited those isolated from the standard diet p > 0 . 05 .


Assuntos
Dieta Hiperlipídica , Resistência Microbiana a Medicamentos , Enterococcus/isolamento & purificação , Fezes/microbiologia , Animais , Aminas Biogênicas/metabolismo , Resistência Microbiana a Medicamentos/efeitos dos fármacos , Enterococcus/patogenicidade , Masculino , Camundongos Endogâmicos ICR , Azeite de Oliva/farmacologia , Filogenia , Técnica de Amplificação ao Acaso de DNA Polimórfico , Fatores de Virulência/metabolismo
7.
Acta Med Port ; 32(9): 600-605, 2019 Sep 02.
Artigo em Português | MEDLINE | ID: mdl-31493364

RESUMO

INTRODUCTION: Infections are a major problem and the presence of drug-resistant microorganisms has significant clinical and economic impact. The present study aims to evaluate the epidemiological profile of "problem" microorganisms isolated in a hospital in the north of Portugal. MATERIAL AND METHODS: All isolated microorganisms were analyzed, between January 2014 and June 2015. Data obtained was then processed using statistical software. RESULTS: We analyzed 8146 microbiological isolations and found a prevalence of 23% of 'problem' microorganisms (in descending order of frequency: Enterococcus, Pseudomonas, Staphylococcus aureus e Streptococcus pneumonia), 57.55% of which isolated in male patients. The most frequent mechanism of drug resistance for the overall sample was the production of extended-spectrum beta-lactamase, and resistance to oxacillin for 'problem' microorganisms. DISCUSSION: In this sample, we observed a much higher prevalence of 'problem' microorganisms than that reported in other countries, which shows the need of improvement of surveillance mechanisms and treatment of these cases. Microorganisms that showed higher resistance were Staphylococcus aureus (resistant to oxacillin) and Enterococcus (resistant to vancomycin). Those were isolated in patients with a higher mean age compared to non-resistant microorganisms. Most of these microorganisms were isolated in hospitalized patients or intermediate and intensive care units, what relates them with healthcare associated infections. CONCLUSION: The prevalence of infection by 'problem' microorganisms during the studied period was 23%. The detection and control of the spread of these microorganisms are paramount due to its impact on health costs, morbidity and survival of patients.


Assuntos
Infecções Bacterianas/microbiologia , Farmacorresistência Bacteriana , Enterococcus/isolamento & purificação , Pseudomonas/isolamento & purificação , Staphylococcus aureus/isolamento & purificação , Streptococcus pneumoniae/isolamento & purificação , Infecções Bacterianas/tratamento farmacológico , Infecções Bacterianas/epidemiologia , Enterococcus/efeitos dos fármacos , Feminino , Humanos , Masculino , Oxacilina/farmacologia , Pseudomonas/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Streptococcus pneumoniae/efeitos dos fármacos , Vancomicina/farmacologia
8.
J Environ Sci Health B ; 54(12): 925-929, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31382830

RESUMO

Dissemination of vancomycin resistance in enterococci has been associated with horizontal transfer of mobile genetic elements. Aim of the study was to evaluate if milk matrix is a suitable environment to support transferability of vancomycin resistance (vanA) gene from clinical vancomycin-resistant Enterococcus faecium to vancomycin-sensitive Enterococcus faecalis. Enterococci strains were firstly screened for the presence of cpd (inducible sex pheromone determinant) gene, vanA and tetL genes (vancomycin and tetracycline resistance markers, respectively) and the gelE (extracellular metalloendopeptidase) gene to define the mating pairs. Based on these selection markers, we investigated the transferability of eight plasmid-borne vanA harbored by E. faecium (vanA+, cpd-, tetL- and gelE-) into two E. faecalis (vanA-, cpd+, tetL + and gelE+) recipient strains in milk matrix. The strains were mated in a 1:1 ratio in 7% reconstituted milk and incubated at 37 °C. Transconjugants emerged from all 16 matings within 2 h of incubation and were evidenced by dual antibiotic resistance (vancomycin and tetracycline). The vancomycin-resistance of trasconjugants was maintained even after ten subsequent passages on nonselective medium. Transconjugants were positive for vanA, tetL and gelE genes. This study indicates milk matrix as suitable environment to support gene exchange between Enterococcus species.


Assuntos
Antibacterianos/farmacologia , Enterococcus/efeitos dos fármacos , Leite/microbiologia , Resistência a Vancomicina , Animais , Proteínas de Bactérias/genética , Bovinos , Conjugação Genética , Enterococcus/classificação , 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 , Contaminação de Alimentos/análise , Infecções por Bactérias Gram-Positivas/microbiologia , Humanos , Testes de Sensibilidade Microbiana , Resistência a Tetraciclina , Vancomicina/farmacologia
9.
BMJ Case Rep ; 12(8)2019 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-31466981

RESUMO

Liver transplant recipients are immunocompromised by the virtue of being on immunosuppressive agents which put them at risk of having infections from unusual and even multiple concomitant pathogens. We present a case of a 39-year-old man who developed septicaemia with Enterococcus casseliflavus, Streptococcus equinus and Klebsiella oxytoca in the setting of perinephric haematoma which resulted following a kidney biopsy performed to evaluate his nephrotic range proteinuria. E. casseliflavus has been known to cause infections in patients with liver disease/cirrhosis; however, simultaneous infection with S. equinus and K. oxytoca along with E. casseliflavus has never been reported earlier in post-transplant state.


Assuntos
Hematoma/etiologia , Nefropatias/complicações , Klebsiella oxytoca/isolamento & purificação , Transplante de Fígado/efeitos adversos , Sepse/microbiologia , Adulto , Aloenxertos/patologia , Antibacterianos/administração & dosagem , Antibacterianos/uso terapêutico , Biópsia/efeitos adversos , Drenagem/métodos , Enterococcus/isolamento & purificação , Hematoma/cirurgia , Humanos , Hospedeiro Imunocomprometido , Imunossupressores/efeitos adversos , Infusões Intravenosas , Rim/diagnóstico por imagem , Rim/microbiologia , Rim/patologia , Nefropatias/diagnóstico por imagem , Nefropatias/microbiologia , Nefropatias/patologia , Cirrose Hepática/complicações , Masculino , Proteinúria/diagnóstico , Sepse/complicações , Sepse/tratamento farmacológico , Streptococcus/isolamento & purificação , Resultado do Tratamento
10.
Mar Pollut Bull ; 146: 11-21, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31426138

RESUMO

A number of Bayesian Networks were developed in order to nowcast and forecast, up to 4 days ahead and in different locations, the likelihood of water quality within the 2018 Commonwealth Games Triathlon swim course exceeding the critical limits for Enterococci and Escherichia coli. The models are data-driven, but the identification of potential inputs and optimal model structure was performed through the parallel contribution of several stakeholders and experts, consulted through workshops. The models, whose main nodes were discretised with a customised discretisation algorithm, were validated over a test set of data and deployed in real-time during the Commonwealth Games in support to a traditional water quality monitoring program. The proposed modelling framework proved to be cost-effective and less time-consuming than process-based models while still achieving high accuracy; in addition, the added value of a continuous stakeholder engagement guarantees a shared understanding of the model outputs and its future deployment.


Assuntos
Enterococcus/crescimento & desenvolvimento , Escherichia coli/crescimento & desenvolvimento , Água Doce/microbiologia , Teorema de Bayes , Enterococcus/isolamento & purificação , Escherichia coli/isolamento & purificação , Qualidade da Água
11.
Int J Antimicrob Agents ; 54(3): 346-350, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31284042

RESUMO

Daptomycin is commonly prescribed in combination with other antibiotics for treatment of enterococcal bacteraemia. Whilst a free drug area under the concentration-time curve to minimum inhibitory concentration (fAUC/MIC) ratio >27.4 is associated with 30-day survival with daptomycin monotherapy, it is unknown whether receipt of other antibiotics affects this threshold. Data were pooled from seven published trials assessing outcomes in daptomycin-treated enterococcal bacteraemia, including patients receiving daptomycin (≥72 h) and any ß-lactam, intravenous aminoglycoside, linezolid, tigecycline and/or vancomycin. Exposures were calculated using a published population pharmacokinetic model based on creatinine clearance, 90% protein binding and daptomycin Etest MIC. The fAUC/MIC threshold predictive of 30-day survival was determined by classification and regression tree analysis. Following pooling of data, 240 adults were included; 137 (57.1%) were alive at 30 days. A majority of patients were immunosuppressed (65.8%) and received a ß-lactam (94.6%). Examining the threshold in low-acuity patients (n = 135) to control for co-morbidities, these patients were more likely to survive when fAUC/MIC >12.3 was achieved (63.2% vs. 20.0%; P = 0.015). The difference remained significant in a multivariable logistic regression model that controlled for infection source and immunosuppression (P = 0.017). This threshold is 2-fold lower than that observed with daptomycin monotherapy. Probabilities of threshold attainment using a 10 mg/kg/day dose were 100% for isolates with MICs ≤ 2 mg/L and 95.2% for a 12 mg/kg/day dose for MICs of 4 mg/L. These data support the use of high-dose daptomycin in combination with another antibiotic for treatment of enterococcal bacteraemia.


Assuntos
Antibacterianos/farmacologia , Antibacterianos/farmacocinética , Bacteriemia/tratamento farmacológico , Daptomicina/farmacologia , Daptomicina/farmacocinética , Enterococcus/efeitos dos fármacos , Infecções por Bactérias Gram-Positivas/tratamento farmacológico , Idoso , Antibacterianos/administração & dosagem , Bacteriemia/microbiologia , Daptomicina/administração & dosagem , Interações de Medicamentos , Quimioterapia Combinada/métodos , Enterococcus/isolamento & purificação , Feminino , Infecções por Bactérias Gram-Positivas/microbiologia , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Análise de Sobrevida , Resultado do Tratamento
12.
Int J Syst Evol Microbiol ; 69(10): 3191-3201, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31339483

RESUMO

Five novel lactic acid bacterial strains, isolated from Chinese traditional pickle juice, were characterised by a polyphasic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, determination of DNA G+C content, average nucleotide identity (ANI) analysis, in silico DNA-DNA hybridisation (isDDH), fatty acid methyl ester (FAME) analysis and an analysis of phenotypic features. Strains 241-2-2T, 63-4T and 190-7T were closely related to the type strains of Enterococcus devriesei, Enterococcus viikkiensis, Enterococcus pseudoavium, Enterococcus xiangfangensis, Enterococcus avium, Enterococcus malodoratus, Enterococcus raffinosus and Enterococcus gilvus, having 99.1-99.9 % 16S rRNA gene sequence similarities. Strain 94-2T was distantly related to the type strains of Enterococcus phoeniculicola, Enterococcus rivorum and Enterococcus faecalis, having 95.2-96.1 % 16S rRNA gene sequence similarities. Strain 85-4T was distantly related to the type strains of Enterococcus casseliflavus, Enterococcus gallinarum, Enterococcus dispar, Enterococcus canintestini, Enterococcus saigonensis, Enterococcus diestrammenae, Enterococcus asini, Enterococcus cecorum and Enterococcus columbae, having 95.7-97.8 % 16S rRNA gene sequence similarities. Less than 91.5 % ANI and 45.3 % isDDH values between strains 241-2-2T, 63-4T, 190-7T, 94-2T, 85-4T and type strains of phylogenetically related species showed that they represent five new species within the genus Enterococcus. Based upon the data of polyphasic characterisation obtained in the present study, five novel species, Enterococcus pingfangensis sp. nov., Enterococcus dongliensis sp. nov., Enterococcus hulanensis sp. nov., Enterococcus nangangensis sp. nov. and Enterococcus songbeiensis sp. nov., are proposed and the type strains are 241-2-2T (=NCIMB 15185T=CCM 8921T=LMG 31181T), 63-4T (=NCIMB 15178T=CCM 8922T), 190-7T (=NCIMB 15200T=CCM 8949T), 94-2T (=NCIMB 15180T=CCM 8920T=LMG 31180T) and 85-4T (=NCIMB 15179T=CCM 8923T=LMG 31183T), respectively.


Assuntos
Enterococcus/classificação , Microbiologia de Alimentos , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Enterococcus/isolamento & purificação , Ácidos Graxos/química , Genes Bacterianos , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
13.
Curr Microbiol ; 76(9): 1010-1019, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31183517

RESUMO

This study addressed the bacteriocin production in 116 lactic acid bacteria isolated from 143 fish and seafood samples. The screening for the production of antibacterial substances allowed for the selection of 16 LAB isolates endowed with inhibitory capability. Bacteriocins (bacLP17 and bacLP18) of two strains, Enterococcus mundtii LP17 and Enterococcus mundtii LP18, respectively, isolated from red mullet and sardine samples, determined large inhibition zones against all the Listeria species. Virulence traits and antibiotic resistances of all producers were verified, and no isolates presented dangerous characteristics, including the two best bacteriocin producers E. mundtii LP17 and E. mundtii LP18, which were subsequently investigated for their potential use in fish and seafood products biopreservation. For both strains, the highest level of bacteriocin production (1280 AU/ml) was recorded when cells were grown at 30 °C in MRS broth at pH ranging from 6.0 to 9.0, and high levels of adsorption of bacteriocins, bacLP17 and bacLP18, to the target cells Listeria monocytogenes were also observed. The results obtained in this study revealed that two strains of E. mundtii originating from seafood exhibited a strong inhibitory activity against L. monocytogenes and may be useful in controlling the growth of this pathogen in the same food products.


Assuntos
Antibacterianos/farmacologia , Bacteriocinas/farmacologia , Enterococcus/química , Enterococcus/isolamento & purificação , Listeria monocytogenes/efeitos dos fármacos , Alimentos Marinhos/microbiologia , Animais , Antibacterianos/biossíntese , Bacteriocinas/biossíntese , Enterococcus/crescimento & desenvolvimento , Enterococcus/metabolismo , Microbiologia de Alimentos , Listeria monocytogenes/crescimento & desenvolvimento , Smegmamorpha/microbiologia
14.
World J Microbiol Biotechnol ; 35(7): 102, 2019 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-31236715

RESUMO

Preparation of curd vary worldwide due to which its taste, texture and impact on human health also differ. In Assam, curd prepared from raw milk (RMC) is preferred over curd prepared from boiled milk (BMC), a tradition believed to have originated from the Mongoloid customs. Microbial diversity of raw milk (RM), boiled milk (BM), RMC and BMC collected from three farms were investigated by culture dependent and independent techniques. Additionally, metabolite profiles of RMC and BMC were studied by gas chromatography and mass spectroscopy. A total of 59 bacterial isolates were identified from the four different dairy products. In RM, lactic acid bacteria such as Lactococcus, Enterococcus, Lactobacillus and Leuconostoc were obtained along with the environmental bacteria like Bacillus, Staphylococcus, Acetobacter, Chryseobacterium, Streptococcus, Acinetobacter, Kocuria, Klebsiella and Macrococcus. Additionally, Prevotella, Oscillospira, Phascolarctobacterium and Akkermansia were also detected in BM by culture independent technique. In RMC and BMC, Lactococcus, Leuconostoc and Lactobacillus were prevalent. RM and RMC shared Enterococcus, Lactococcus, Streptococcus and Acinetobacter as common bacterial genera. However, no bacterial genus was common in BM and BMC. The correlation analysis revealed that Lactobacillus was negatively correlated to other bacterial genera. Oligotyping analysis revealed that Lactobacillus brevis and L.fermentum were abundant in RMC and BMC, respectively. In metabolomic study, ascorbic acid, dodecanoic acid and hexadecanoic acid were found to be significantly higher in RMC. Presence of different types of probiotics in these curds samples opens a new avenue to understand their effects on human health.


Assuntos
Bactérias/isolamento & purificação , Biodiversidade , Fermentação , Leite/microbiologia , Animais , Bactérias/metabolismo , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , Enterococcus/isolamento & purificação , Enterococcus/metabolismo , Manipulação de Alimentos , Microbiologia de Alimentos , Sequenciamento de Nucleotídeos em Larga Escala , Lactobacillales , Lactobacillus/isolamento & purificação , Lactobacillus/metabolismo , Lactococcus/isolamento & purificação , Lactococcus/metabolismo , Leuconostoc/isolamento & purificação , Leuconostoc/metabolismo , Metagenômica , Análise Multivariada , RNA Ribossômico 16S/genética , RNA Ribossômico 16S/isolamento & purificação
15.
Lett Appl Microbiol ; 69(3): 204-211, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31250457

RESUMO

Despite of the beneficial relevance of several lactic acid bacteria (LAB) in the food industry, micro-organisms belonging to this group can determine spoilage in food products and carry a number of virulence and antibiotic resistance-related genes. This study aimed on the characterization of beneficial and safety aspects of five bacteriocinogenic LAB strains (Lactobacillus curvatus 12-named L. curvatus UFV-NPAC1), L. curvatus 36, Weissela viridescens 23, W. viridescens 31 and Lactococcus garvieae 36) isolated from an artisanal Brazilian calabresa, a traditional meat sausage. Regarding their beneficial aspects, all tested isolates were positive for mub, while EF226-cbp, EF1249-fbp and EF2380-maz were detected in at least one tested strain; none of the isolates presented map, EFTu or prgB. However, evaluated strains presented a variable pattern of virulence-related genes, but none of the strains presented gelE, cylA, efsA, cpd, int-Tn or sprE. Moreover, other virulence-related genes evaluated in this study were detected at different frequencies. L. curvatus 12 was generated positive results for ace, ccf, int, ermC, tetL, aac(6')-Ie-aph(2″)-Ia, aph(2″)-Ib, aph(2″)-Ic, bcrB, vanB and vanC2; L. curvatus 36: hyl, asa1, esp, int, ermC, tetK, aph(3')-IIIa, aph(2'')-Ic and vanC2; L. garvieae 32: asa1, ant(4')-Ia, aph(2'')-Ib, catA, vanA and vanC1; W. viridescens 23: esp, cob, ermB, aph(3')-IIIa, aph(2'')-Ic, vanA, vanB and vanC2; W. viridescens 31: hyl, esp, ermC, aph(3')-IIIa, aph(2'')-Ib, aph(2'')-Ic, catA, vanA and vanB. Despite presenting some beneficial aspects, the presence of virulence and antibiotic resistance genes jeopardize their utilization as starter or biopreservatives cultures in food products. Considering the inhibitory potential of these strains, an alternative would be the use of their bacteriocins as semi-purified or pure technological preparation. SIGNIFICANCE AND IMPACT OF THE STUDY: The food industry has a particular interest in using bacteriocinogenic lactic acid bacteria (LAB) as starter, probiotics and/or biopreservatives in different food products. Characterization of additional beneficial features is important to identify new, multifunctional potential probiotic strains. However, these strains can only be applied in food products only after being properly characterized according their potential negative aspects, such as virulence and antibiotic resistance genes. A wide characterization of beneficial and safety aspects of bacteriocinogenic LAB is determinant to guide the proper utilization of these strains, or their purified bacteriocins, by the food industry.


Assuntos
Antibacterianos/farmacologia , Bacteriocinas/biossíntese , Lactobacillales/genética , Fatores de Virulência/genética , Brasil , Resistência Microbiana a Medicamentos/genética , Enterococcus/isolamento & purificação , Microbiologia de Alimentos , Lactobacillales/isolamento & purificação , Lactobacillus/efeitos dos fármacos , Lactococcus/isolamento & purificação , Produtos da Carne/microbiologia , Virulência
16.
Mar Pollut Bull ; 144: 36-47, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31180005

RESUMO

Non-point sources of sewage-related pollution in tropical marine waters are difficult to ascertain. Enterococci (ENT) are widely used as indicators of human waste but their efficacy in tropical waters is highly debated due to natural presence in tropical soils. Clostridium perfringens (CP) is often used as a secondary indicator of fecal contamination because its presence indicates sewage, and in tropical waters environmental sources are unlikely. We analyzed a 27-year dataset containing over 29,000 samples collected by the State of Hawaii, to determine a proposed CP standard for detecting human sewage, which has applicability throughout tropical marine waters globally. Measured ENT concentrations were highly correlated with turbidity. In three instances, sewage contamination was not detected by ENT samples alone, and impairments from non-point pollution may be highly misinformed in Hawaii. The EPA should examine relationships between CP and human health and implement CP as the primary FIB in tropical marine waters.


Assuntos
Clostridium perfringens/isolamento & purificação , Enterococcus/isolamento & purificação , Monitoramento Ambiental/métodos , Água do Mar/microbiologia , Esgotos/microbiologia , Microbiologia da Água/normas , Fezes/microbiologia , Hawaii , Humanos , Reprodutibilidade dos Testes
17.
Nat Microbiol ; 4(11): 1826-1831, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31209308

RESUMO

Recent work has highlighted the importance of confounder control in microbiome association studies1,2. For instance, multiple pathologies previously linked to gut ecosystem dysbiosis display concomitant changes in stool consistency3-6, a major covariate of microbiome variation2,7. In those cases, observed microbiota alterations could largely reflect variation in faecal water content. Moreover, stool moisture variation has been linked to fluctuations in faecal microbial load, inducing artefacts in relative abundance profile analyses8,9. Hence, the identification of associations between the gut microbiota and specific disease manifestations in pathologies with complex aetiologies requires a deconfounded, quantitative assessment of microbiome variation. Here, we revisit a disease association microbiome data set comprising 106 patients with primary sclerosing cholangitis (PSC) and/or inflammatory bowel disease10. Assessing quantitative taxon abundances9, we study microbiome alterations beyond symptomatic stool moisture variation. We observe an increased prevalence of a low cell count Bacteroides 2 enterotype across the pathologies studied, with microbial loads correlating inversely with intestinal and systemic inflammation markers. Quantitative analyses allow us to differentiate between taxa associated with either intestinal inflammation severity (Fusobacterium) or cholangitis/biliary obstruction (Enterococcus) among previously suggested PSC marker genera. We identify and validate a near-exclusion pattern between the inflammation-associated Fusobacterium and Veillonella genera, with Fusobacterium detection being restricted to Crohn's disease and patients with PSC-Crohn's disease. Overall, through absolute quantification and confounder control, we single out clear-cut microbiome markers associated with pathophysiological manifestations and disease diagnosis.


Assuntos
Bactérias/classificação , Colangite Esclerosante/microbiologia , Doenças Inflamatórias Intestinais/microbiologia , Metagenômica/métodos , Bactérias/genética , Bactérias/isolamento & purificação , Bacteroides/isolamento & purificação , Enterococcus/isolamento & purificação , Fezes/microbiologia , Feminino , Fusobacterium/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade
18.
BMC Infect Dis ; 19(1): 531, 2019 Jun 17.
Artigo em Inglês | MEDLINE | ID: mdl-31208347

RESUMO

BACKGROUND: Clinical microbiology laboratories are asked to process large numbers of urine specimens for culture, but only 20-40% of them are positive. Therefore, a rapid, reliable screening method is necessary to speed up the reporting of a negative result. In this study, we evaluated the iQ200/iChem workstation, which is a combination of digital imaging software and a strip reader to predict negative urine culture. METHOD: A total of 1942 urine specimens were processed through both culture and iQ200/ iChem workstation. We analyzed the performance using two definition of positive urine culture; one or two potential uropathogens at a concentration of ≥105 CFU/ml and ≥ 104 CFU/ml. We assessed combinations of parameters (ASP; all small particles, WBC; leukocyte, BACT; bcteria, LE; leukocyte esterase) applying various cut-offs which can achieve the negative predictive value (NPV) ≥97% and culture reduction rate ≥ 50%. RESULTS: The culture positive rate was 12.8 and 18.4% applying the criteria of ≥105 CFU/ml and ≥ 104 CFU/ml, respectively. The area under the curve (AUC) of each parameter for ≥105 CFU/ml / ≥104 CFU/ml bacteriuria was 795 /0.719 for WBC, 0.722 / 0.701 for ASP and 0.740 /0.704 for bacteria. Therefore, we investigated the combination of the parameters. With the fixed parameter of BACT≥1/HPF and positive LE, the combinations of WBC ≥ 4/HPF and ASP ≥8500/µl or WBC ≥ 6/HPF and ASP≥5500/µl showed good performance for detecting ≥105 CFU/ml uropathogen. The ranges of sensitivity, specificity, negative predictive value and culture reduction rate were 91.5-92.3%, 49.8-52.6%, 97.7-97.9% and 50.4-53.0%, respectively. However, none of the combined setting yielded acceptable range of NPV for detecting ≥104 CFU/ml uropathogen (NPV 92.9-94.9%). Enterococcus spp. was the most common uropathogen causing the false negative results (55.7%), and also the main pathogen among the positive culture of 104-5 CFU/ml bacteriuria (45%). CONCLUSIONS: iQ200/iChem workstation was excellent in detection of ≥105 CFU/ml uropathogen, but unsatisfactory in detection of 104-5 CFU/ml uropathogen and Enterococcus spp. It can be useful for screening of urine specimens to reduce bacterial culture. However, notice from clinician will be necessary for specimens from the patients with high risk for UTI, such as pregnant woman, infant, elderly or immune compromised patients.


Assuntos
Urinálise/instrumentação , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Bacteriúria/diagnóstico , Hidrolases de Éster Carboxílico/análise , Criança , Pré-Escolar , Enterococcus/isolamento & purificação , Feminino , Humanos , Processamento de Imagem Assistida por Computador , Lactente , Recém-Nascido , Leucócitos , Masculino , Programas de Rastreamento , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Software , Urinálise/métodos , Infecções Urinárias/microbiologia , Urina/microbiologia , Adulto Jovem
19.
World J Microbiol Biotechnol ; 35(7): 100, 2019 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-31222403

RESUMO

The spontaneously fermented curdled milk product from Burkina Faso, lait caillé is prepared by traditional processing from raw unpasteurised milk. The fermentation lasts 1-3 days. This study aims to identify the predominant microbiota involved in lait caillé fermentation from cow milk. A survey on lait caillé end-products from local markets showed pH ranges of 3.5 to 4.2. Counts of total lactic acid bacteria (LAB) were 7.8 ± 0.06 to 10.0 ± 0.03 log CFU/g and yeast counts were 5.3 ± 0.06 to 8.7 ± 0.01 log CFU/g, together with considerate amounts of Enterobacteriaceae < 3.00 to 8.4 ± 0.14 log CFU/g. Sampling throughout the entire fermentation of lait caillé was performed at a traditional house-hold production site. A drop in pH from 6.7 ± 0.01 at 0 h to 4.3 ± 0.08 in the end-product (59 h) was found. Total LAB counts increased to 8.6 ± 0.02 log CFU/g in the end-product, while yeast and Enterobacteriaceae counts reached 6.4 ± 0.11 and 6.7 ± 0.00 log CFU/g, respectively. LAB and yeasts isolated during the fermentation were clustered by (GTG)5 repetitive-PCR fingerprinting followed by 16S and 26S rRNA gene sequencing, respectively. Microbial successions were observed with Leuconostoc mesenteroides being the predominant LAB followed by Pediococcus pentosaceus and Weissella paramesenteroides at the onset, while Lactococcus lactis and Enterococcus spp. where the predominant LAB after 7 h of fermentation. During the first 18 h Candida parapsilosis was the dominant yeast species, while from 35 h to the end-product, Saccharomyces cerevisiae predominated. The microbial safety risk pointed out in this study, showed the need for implementation of good manufacturing practices including pasteurisation and use of well-defined starter cultures.


Assuntos
Produtos Fermentados do Leite/microbiologia , Microbiota/genética , Burkina Faso , Contagem de Colônia Microbiana , Enterobacteriaceae/genética , Enterobacteriaceae/isolamento & purificação , Enterococcus/genética , Enterococcus/isolamento & purificação , Fermentação , Manipulação de Alimentos , Microbiologia de Alimentos , Concentração de Íons de Hidrogênio , Lactobacillales/genética , Lactobacillales/isolamento & purificação , Lactococcus lactis/genética , Lactococcus lactis/isolamento & purificação , RNA Ribossômico/genética , RNA Ribossômico/isolamento & purificação , RNA Ribossômico 16S/genética , RNA Ribossômico 16S/isolamento & purificação , Análise de Sequência de RNA , Leveduras/genética , Leveduras/isolamento & purificação
20.
Int J Syst Evol Microbiol ; 69(8): 2506-2513, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31204971

RESUMO

A Gram-stain-positive and catalase negative coccus, designated strain Gos25-1T, isolated from a cotton flower (Gossypium hirsutum L.) collected from Khao Wong district, Kalasin province, Thailand. The taxonomic position of this strain was systematically studied based upon polyphasic taxonomic methods. The strain was facultatively anaerobic and produced l-lactic acid from glucose. The predominant cellular fatty acids were the straight-chain fatty acids C18 : 1ω9c and C16 : 0. According to 16S rRNA and phenylalanyl-tRNA synthase alpha subunit (pheS) gene sequence similarity, this strain was closely related to Enterococcus pallens NBRC 100697T, E. hermanniensis CIP 108559T, E. avium NBRC 100477T and E. raffinosus NBRC 100492T with 98.9-99.1 % and 77.0-82.0 % sequence similarities, respectively. Phylogenetic analysis indicated that strain Gos25-1T was clearly distinguished from closely related species of the genus Enterococcus. Draft genome of Gos25-1T had a size of 3.99 Mb which was contained 3788 coding sequences with in silico G+C content of 42.4 mol%. The ANIb and a digital DNA-DNA hybridisation (dDDH) values between strain Gos25-1T and the closest related species, E. pallens NBRC 100697T were 73.65 and 21.10 %, respectively. According to polyphasic characterisation, this strain represents a novel species of the genus Enterococcus, for which the name Enterococcus florum sp. nov. is proposed. The type strain is Gos25-1T (=CIP 110956T=LMG 29007T=NBRC 111461T=TISTR 2382T).


Assuntos
Enterococcus/classificação , Flores/microbiologia , Gossypium/microbiologia , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Enterococcus/isolamento & purificação , Ácidos Graxos/química , Genes Bacterianos , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Tailândia
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