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Id: biblio-974302
Autor: Barreiro, Juliana R; Gonçalves, Juliano L; Grenfell, Rafaella; Leite, Renata F; Juliano, Luiz; Santos, Marcos V.
Título: Direct identification of bovine mastitis pathogens by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry in pre-incubated milk
Fonte: Braz. j. microbiol;49(4):801-807, Oct.-Dec. 2018. tab, graf.
Idioma: en.
Projeto: FAPESP; . Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University.
Resumo: ABSTRACT The present study aimed to compare two MALDI-TOF identification methods [(a) direct sample identification after pre-incubation; or (b) use of bacteria isolated on pre-culture)] to standard, traditional bench microbiology. A total of 120 quarter milk samples from 40 Holstein lactating cows were screened based on culture-positive results obtained by microbiological culture (reference method) with the following numbers of quarters positive per cow: 4 cows with 1, 8 cows with 2, 12 cows with 3 and 16 cows with 4 infected quarters per cow. For direct identification method, quarter milk samples (n = 120) were skimmed by centrifugation (10,000 × g/10 min) and pre-incubated at 37 ºC for 12 h. After pre-incubation, quarter milk samples were submitted to total bacterial count by flow cytometry and for a preparation protocol for bacterial ribosomal protein extraction followed by MALDI-TOF MS analysis. The direct MALDI-TOF MS identification method compared to microbiological culture correctly identified isolates of coagulase-negative Staphylococci (27.2%), Streptococcus agalactiae (21.8%), Staphylococcus aureus (14.2%), and Streptococcus uberis (5.2%). The pre-incubation protocol of milk samples, associated to the direct identification method by MALDI-TOF MS, did not increase the identification at species level (score >2.0) of pathogens causing subclinical mastitis in comparison to the method without previous incubation.
Descritores: Staphylococcus/isolamento & purificação
Streptococcus/isolamento & purificação
Técnicas de Tipagem Bacteriana/métodos
Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos
Leite/microbiologia
Mastite Bovina/microbiologia
-Staphylococcus/genética
Staphylococcus/química
Streptococcus/genética
Streptococcus/química
Leite/química
Mastite Bovina/fisiopatologia
Limites: Animais
Feminino
Lactente
Bovinos
Responsável: BR1.1 - BIREME


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Vasconcellos, Silvio Arruda
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Id: biblio-974290
Autor: Karcher, Daniel; Grenfell, Rafaella C; Moreno, Andrea Micke; Moreno, Luisa Zanolli; Vasconcellos, Silvio Arruda; Heinemann, Marcos B; Almeida Junior, Joao N. de; Juliano, Luiz; Juliano, Maria A.
Título: Identification of pathogenic and nonpathogenic Leptospira species of Brazilian isolates by Matrix Assisted Laser Desorption/Ionization and Time Flight mass spectrometry
Fonte: Braz. j. microbiol;49(4):900-908, Oct.-Dec. 2018. tab, graf.
Idioma: en.
Projeto: FAPESP; . CNPq.
Resumo: ABSTRACT Matrix Assisted Laser Desorption/Ionization and Time of Flight mass spectrometry (MALDI-TOF MS) is a powerful tool for the identification of bacteria through the detection and analysis of their proteins or fragments derived from ribosomes. Slight sequence variations in conserved ribosomal proteins distinguish microorganisms at the subspecies and strain levels. Characterization of Leptospira spp. by 16S RNA sequencing is costly and time-consuming, and recent studies have shown that closely related species (e.g., Leptospira interrogans and Leptospira kirschneri) may not be discriminated using this technology. Herein, we report an in-house Leptospira reference spectra database using Leptospira reference strains that were validated with a collection of well-identified Brazilian isolates kept in the Bacterial Zoonosis Laboratory at the Veterinary Preventive Medicine and Animal Health Department at Sao Paulo University. In addition, L. interrogans and L. kirschneri were differentiated using an in-depth mass spectrometry analysis with ClinProTools™ software. In conclusion, our in-house reference spectra database has the necessary accuracy to differentiate pathogenic and non-pathogenic species and to distinguish L. interrogans and L. kirschneri.
Descritores: Técnicas de Tipagem Bacteriana/métodos
Espectrometria de Massas em Tandem/métodos
Leptospira/isolamento & purificação
Leptospirose/microbiologia
-Brasil
DNA Bacteriano/genética
RNA Ribossômico 16S/genética
Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos
Leptospira/classificação
Leptospira/genética
Leptospira/química
Limites: Seres Humanos
Tipo de Publ: Estudos de Avaliação
Responsável: BR1.1 - BIREME


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Id: biblio-974336
Autor: Zhao, Junfeng; Zhang, Chong; Lu, Zhaoxin.
Título: Differential proteomics research of Bacillus amyloliquefaciens and its genome-shuffled saltant for improving fengycin production
Fonte: Braz. j. microbiol;49(supl.1):166-177, 2018. tab, graf.
Idioma: en.
Projeto: National Natural Science Foundation of China.
Resumo: Abstract In the previous study, we used genome shuffling to improve fengycin production of the original strain Bacillus amyloliquefaciens ES-2-4. After two rounds of genome shuffling, a high-yield recombinant FMB72 strain that exhibited 8.30-fold increase in fengycin production was obtained. In this study, comparative proteomic analysis of the parental ES-2-4 and genome-shuffled FMB72 strains was conducted to examine the differentially expressed proteins. In the shuffled strain FMB72, 50 differently expressed spots (p < 0.05) were selected to be excised and analyzed using Matrix-Assisted Laser Desorption/Ionization Time of Flight/Time of Flight Mass Spectrometry, and finally 44 protein spots were confidently identified according to NCBI database. According to clusters of orthologous groups (COG) functional category analysis and related references, the differentially expressed proteins could be classified into several functional categories, including proteins involved in metabolism, energy generation and conversion, DNA replication, transcription, translation, ribosomal structure and biogenesis, cell motility and secretion, signal transduction mechanisms, general function prediction. Of the 44 identified proteins, signaling proteins ComA and Spo0A may positively regulate fengycin synthesis at transcriptional level. Taken together, the present study will be informative for exploring the exact roles of ComA and Spo0A in fengycin synthesis and explaining the molecular mechanism of fengycin synthesis.
Descritores: Proteínas de Bactérias/metabolismo
Lipopeptídeos/biossíntese
Bacillus amyloliquefaciens/genética
Bacillus amyloliquefaciens/metabolismo
-Proteínas de Bactérias/genética
Proteínas de Bactérias/química
Genoma Bacteriano
Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
Embaralhamento de DNA
Proteômica
Bacillus amyloliquefaciens/química
Responsável: BR1.1 - BIREME


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Id: biblio-828202
Autor: Jha, Sujata S; Joshi, Sanket J; S. J., Geetha.
Título: Lipopeptide production by Bacillus subtilis R1 and its possible applications
Fonte: Braz. j. microbiol;47(4):955-964, Oct.-Dec. 2016. tab, graf.
Idioma: en.
Resumo: Abstract The possible application of a bacterial strain - Bacillus subtilis R1, isolated from an oil contaminated desert site in India, as biocontrol agent and its biosurfactant in microbial enhanced oil recovery are discussed. The biosurfactant production in minimal medium was carried out at different temperatures and salt concentrations, where it produced an efficient biosurfactant at 30-45 °C and in presence of up to 7% salt. It significantly reduced the surface tension from 66 ± 1.25 mN/m to 29 ± 0.85 mN/m within 24 h. In order to enhance the biosurfactant production, random mutagenesis of B. subtilis R1 was performed using chemical mutagen - ethyl methanesulfonate. Majority of the isolated 42 mutants showed biosurfactant production, but the difference was statistically insignificant as compared with parent strain R1. Therefore none of the mutants were selected for further study, and only parent strain R1 was studied. The biosurfactant was quite stable under harsh conditions for up to 10 days. The biosurfactant was extracted and characterized as similar to the lipopeptide group - surfactins and fengycin. The crude oil displacement experiments using biosurfactant broth in sand pack glass columns showed 33 ± 1.25% additional oil recovery. The strain also showed inhibition of various plant pathogenic fungi on potato dextrose agar medium.
Descritores: Bacillus subtilis/metabolismo
Lipopeptídeos/biossíntese
-Tensoativos/metabolismo
Tensoativos/farmacologia
Bacillus subtilis/classificação
Bacillus subtilis/genética
RNA Ribossômico 16S/genética
Testes de Sensibilidade Microbiana
Mutagênese
Espectroscopia de Infravermelho com Transformada de Fourier
Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
Lipopeptídeos/farmacologia
Engenharia Metabólica
Concentração de Íons de Hidrogênio
Antifúngicos/metabolismo
Antifúngicos/farmacologia
Responsável: BR1.1 - BIREME


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Id: lil-780818
Autor: AlMogbel, Mohammed Suliman.
Título: Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry for identification of Clostridium species isolated from Saudi Arabia
Fonte: Braz. j. microbiol;47(2):410-413, Apr.-June 2016. tab, graf.
Idioma: en.
Resumo: Abstract The aim of this study was to identify different Clostridium spp. isolated from currency notes from the Ha’il region of Saudi Arabia in September 2014 using MALDI–TOF-MS. Clostridium spp. were identified by Bruker MALDI–TOF-MS and compared with VITEK 2. The confirmation of the presence of different Clostridium spp. was performed by determining the sequence of the 16S ribosomal RNA gene. In this study, 144 Clostridium spp. were isolated. Among these specimens, MALDI–TOF-MS could identify 88.8% (128/144) of the isolates to the species level and 92.3% (133/144) to the genus level, whereas, VITEK 2 identified 77.7% of the (112/144) isolates. The correct identification of the 144 isolates was performed by sequence analysis of the 500 bp 16S rRNA gene. The most common Clostridium spp. identified were Clostridium perfringens (67.36%), Clostridium subterminale (14.58%), Clostridium sordellii (9%) and Clostridium sporogenes (9%). The results of this study demonstrate that MALDI–TOF-MS is a rapid, accurate and user friendly technique for the identification of Clostridium spp. Additionally, MALDI–TOF-MS has advantages over VITEK 2 in the identification of fastidious micro-organisms, such as Clostridium spp. Incorporating this technique into routine microbiology would lead to more successful and rapid identification of pathogenic and difficult to identify micro-organisms.
Descritores: Clostridium/isolamento & purificação
Clostridium/química
Espectrometria de Massas em Tandem/métodos
-Arábia Saudita
Técnicas de Tipagem Bacteriana/métodos
Clostridium/classificação
Clostridium/genética
Infecções por Clostridium/microbiologia
Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos
Limites: Seres Humanos
Responsável: BR1.1 - BIREME


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Moreira, Beatriz Meurer
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Id: biblio-839355
Autor: Rodrigues, Naiara Miranda Bento; Bronzato, Greiciane França; Santiago, Gabrielli Stefaninni; Botelho, Larissa Alvarenga Batista; Moreira, Beatriz Meurer; Coelho, Irene da Silva; Souza, Miliane Moreira Soares de; Coelho, Shana de Mattos de Oliveira.
Título: The Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) identification versus biochemical tests: a study with enterobacteria from a dairy cattle environment
Fonte: Braz. j. microbiol;48(1):132-138, Jan.-Mar. 2017. tab.
Idioma: en.
Projeto: National Council for Scientific and Technological Development.
Resumo: Abstract Mastitis adversely affects milk production and in general cows do not regain their full production levels post recovery, leading to considerable economic losses. Moreover the percentage decrease in milk production depends on the specific pathogen that caused the infection and enterobacteria are responsible for this greater reduction. Phenotypic tests are among the currently available methods used worldwide to identify enterobacteria; however they tend to misdiagnose the species despite the multiple tests carried out. On the other hand The Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) technique has been attracting attention for its precise identification of several microorganisms at species level. In the current study, 183 enterobacteria were detected in milk (n = 47) and fecal samples (n = 94) from cows, and samples from water (n = 23) and milk lines (n = 19). All these samples were collected from a farm in Rio de Janeiro with the specific purpose of presenting the MALDI-TOF MS technique as an efficient methodology to identify Enterobacteriaceae from bovine environments. The MALDI-TOF MS technique results matched the biochemical test results in 92.9% (170/183) of the enterobacteria species and the gyrB sequencing confirmed 100% of the proteomic technique results. The amino acid decarboxylation test made the most misidentifications and Enterobacter spp. was the most misidentified genus (76.9%, 10/13). These results aim to clarify the current biochemical errors in enterobacteria identification, considering isolates from a bovine environment, and show the importance for more careful readings of phenotypic tests which are often used in veterinary microbiology laboratories.
Descritores: Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
Enterobacteriaceae/classificação
Enterobacteriaceae/metabolismo
-Fenótipo
Bovinos
Análise de Sequência de DNA
DNA Girase/genética
Proteômica/métodos
Leite/microbiologia
Enterobacteriaceae/isolamento & purificação
Genes Bacterianos
Mastite Bovina/diagnóstico
Mastite Bovina/microbiologia
Limites: Animais
Feminino
Responsável: BR1.1 - BIREME


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Id: lil-621523
Autor: Pasternak, Jacyr.
Título: New methods of microbiological identification using MALDI-TOF / Novas metodologias de identificação de micro-organismos: MALDI-TOF
Fonte: Einstein (Säo Paulo);10(1):118-119, jan.-mar. 2012.
Idioma: en; pt.
Resumo: Rapid diagnosis of pathogens is decisive to guarantee adequate therapy in infections: culture methods are precise and sensitive, but rather slow. New resources are available to enable faster diagnosis, and the most promising is MALDI-TOF technology: mass spectrometry applied to microbiological diagnosis. Times as fast as 10 to 15 minutes to etiological diagnosis are possible after a positive blood culture result. We hope to have this technology in our laboratory, ANVISA permitting and improving their very slow rate of doing things... MALDI-TOF is basically putting a sample of culture or an enriched suspension of the probable pathogen over a small spot with a matrix and vaporizing it with a laser pulse: the products are aspired into a chamber, ionized and reach detectors at variable times: the detectors show time of arrival and quantity of the product, and each pathogen has its characteristic spectrum analyzed by a software.

O diagnóstico rápido de patógenos é crucial para a terapêutica adequada. O método clássico de cultura é preciso e sensível, mas demorado; novos métodos permitem um diagnóstico muito mais rápido, que pode ser tão precoce como 15 minutos após obter um material enriquecido. Em uma hemocultura, por exemplo, é possível saber o agente etiológico nesse prazo, assim que a cultura se mostrar positiva. Esse novo método é o MALDI-TOF, uma aplicação da espectrometria de massa à microbiologia: o material é colocado em uma placa com matriz e bombardeado com um laser que o evapora; um sistema ioniza e aspira o material volatilizado, que chega a detectores, os quais registram o tempo em que a substância chega ao detector e sua quantidade. Cada patógeno tem um espectro característico que é analisado por um software.
Descritores: Técnicas de Tipagem Bacteriana
Técnicas de Tipagem Micológica
Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
-Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
Fatores de Tempo
Limites: Seres Humanos
Responsável: BR1.1 - BIREME


  8 / 44 LILACS  
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Araújo, Flábio R
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Id: biblio-895409
Autor: Bier, Daniele; Tutija, Juliane F; Pasquatti, Taynara N; Oliveira, Tayná L; Araújo, Flábio R; Verbisck, Newton V.
Título: Identificação por espectrometria de massa MALDI-TOF de Salmonella spp. e Escherichia coli isolados de carcaças bovinas / MALDI-TOF mass spectrometry identification of Salmonella spp. and Escherichia coli isolated from bovine carcasses
Fonte: Pesqui. vet. bras = Braz. j. vet. res;37(12):1373-1379, dez. 2017. tab, graf.
Idioma: pt.
Projeto: Fundação de Apoio ao Desenvolvimento do Ensino, Ciência e Tecnologia do Estado de Mato Grosso do Sul; . Embrapa Gado de Corte.
Resumo: O objetivo deste trabalho foi introduzir a técnica de espectrometria de massa com fonte de ionização e dessorção a laser assistida por matriz e analisador de tempo-de-voo (MALDI-TOF) para incrementar o método tradicional microbiológico na detecção de Salmonella spp. e Escherichia coli em carcaças bovinas. Foram avaliadas 270 amostras de 90 carcaças de bovinos. Para isolamento de Salmonella spp. e E. coli, foram utilizadas, respectivamente, as metodologias descritas na ISO 6579:2002 e no Compendium of Methods for the Microbiological Examination of Foods. As análises por MALDI-TOF foram realizadas a partir de isolados cultivados em ágar nutriente ou em caldo triptona de soja, provenientes das amostras com características bioquímicas positivas (n=7), inconclusivas (n=4) e negativas (n=85) para Salmonella spp. e bioquímicas positivas (n=37) e negativas (n=85) para E. coli. Os perfis de massas foram adquiridos com o espectrômetro de massas MALDI-TOF Autoflex III SmartBeam e os espectros brutos foram processados usando o programa MALDI Biotyper (Bruker Daltonics). De acordo com a identificação preliminar, com base na morfologia das colônias e nas reações bioquímicas, sete isolados foram considerados positivos para Salmonella spp. Através do MALDI Biotyper, esses sete isolados foram classificados como pertencentes ao gênero Salmonella e, além disso, identificados como S. enterica. Quatro isolados que apresentaram características fenotípicas não usuais e resultados inconclusivos nos testes bioquímicos para Salmonella foram identificados como pertencentes aos gêneros Citrobacter e Proteus após análise por MALDI. Para E. coli, 37 amostras foram positivas pelos testes bioquímicos da espécie, o que foi confirmado por MALDI Biotyper. A metodologia MALDI-TOF permitiu a rápida confirmação da identidade de Salmonella spp. e E. coli, podendo ser utilizada para detecção desses microrganismos em isolados bacterianos de carcaças bovinas.(AU)

The aim of this study was to introduce matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry to improve the traditional microbiological method for the detection of Salmonella spp. and Escherichia coli in beef carcasses. Two hundred seventy samples from 90 beef carcasses were evaluated. The methodologies described in ISO 6579:2002 and in the Compendium of Methods for the Microbiological Examination of Foods were used for Salmonella spp. and E. coli isolation, respectively. MALDI-TOF analysis were performed on tryptone soya broth suspension isolates or directly from nutrient agar colonies, from the positive, inconclusive or negative biochemically tested samples for Salmonella and E. coli. Mass profiles were acquired on an Autoflex III SmartBeam MALDI-TOF mass spectrometer and the raw spectra were processed using the MALDI Biotyper software (Bruker Daltonics). According to the preliminary identification based on colony morphology and the biochemical reactions, seven isolates were positive for Salmonella spp. Through MALDI Biotyper these seven isolates were also classified as belonging to the genus Salmonella and further identified as S. enterica. Four isolates showing unusual phenotypic characteristics and inconclusive results in biochemical tests for Salmonella were identified as belonging to Citrobacter and Proteus genera after MALDI analysis. Regarding Escherichia coli, 37 were positive for species biochemical testing which MALDI Biotyper confirmed. MALDI-TOF methodology allowed rapid Salmonella spp. and E. coli identity confirmation and may be used to detect these microrganisms within bacterial isolates from beef carcasses.(AU)
Descritores: Salmonella
Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/veterinária
Escherichia coli
Carne/microbiologia
-Espectrometria de Massas/veterinária
Matadouros
Enterobacteriaceae
Limites: Animais
Bovinos
Responsável: BR68.1 - Biblioteca Virginie Buff D'Ápice


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Id: biblio-1047051
Autor: Araújo, Max Roberto Batista; Wolf, Vanessa; Seabra, Luisa Ferreira.
Título: Identification of Chromobacterium violaceum by MALDI-TOF mass spectrometry in blood culture
Fonte: Clin. biomed. res;38(3):308-310, 2018.
Idioma: en.
Resumo: Report of Chromobacterium violaceum isolation from blood culture. Identification by MALDI-TOF mass spectrometry. Relevant report due to the site affected, infection severity, and importance of correct and rapid identification for a successful treatment and lower risk of morbidity and mortality. (AU)
Descritores: Chromobacterium/patogenicidade
-Sepse/diagnóstico
Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
Hemocultura/métodos
Limites: Seres Humanos
Masculino
Meia-Idade
Tipo de Publ: Relatos de Casos
Responsável: BR18.1 - Biblioteca FAMED/HCPA


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Id: biblio-1025675
Autor: Araújo, Max Roberto Batista; Santos, Elaine Gomes de Morais; Wolf, Vanessa; Seabra, Luisa Ferreira.
Título: Identification of Cryptococcus neoformans by MALDI-TOF mass spectrometry in blood culture
Fonte: Clin. biomed. res;38(2), 2018.
Idioma: en.
Resumo: This is a report of isolation of Cryptococcus neoformans from blood culture. Identification was conducted by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. The relevance of this report is related to the site affected, the severity of the infection, and the importance of correct and rapid identification of the causative pathogen for a successful treatment and for reducing the risk of morbidity and mortality. (AU)
Descritores: Criptococose/diagnóstico
Cryptococcus neoformans/patogenicidade
Cryptococcus neoformans/virologia
-Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
Hemocultura/métodos
Limites: Seres Humanos
Feminino
Idoso
Tipo de Publ: Relatos de Casos
Responsável: BR18.1 - Biblioteca FAMED/HCPA



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