Método rápido de extração de DNA de bactérias

Desenvolveu-se um método rápido para extração de DNA de bactérias, que ao contrário de outros métodos, não requer o uso de enzimas, como lisozima e proteinase K, previamente, utilizado-se o carbonato de silício (carborundum) como agente físico para efetuar a quebrar da parede celular da bactéria. Com este método conseguiu-se extrair DNA bacteriano num menor tempo, além de mais rápido, ele mostrou-se mais simples e econômico, quando comparado aos métodos convencionais. O DNA obtido pode ser utilizado para diversas finalidades relacionadas ao DNA de bactérias, obtendo-se uma quantidade razoável de DNA, que varia de 725 µg/mL a 1170 µg/mL por cada 0,1 g de célula bacteriana, com ótima qualidade.

Avaliação, identificação e atividade enzimática de bactérias psicotróficas presentes no leite cru refrigerado

Extended storage of refrigerated milk can lead to reduced quality of raw and processed milk, which is a consequence of the growth and metabolic activities of psychrotrophic bacteria, able to grow under 7oC or lower temperatures. Although most of these microorganisms are destroyed by heat treatment, some have the potential to produce termoresistant proteolytic and lipolytic enzymes that can survive even UHT processing and reduce the processed products quality. Recently, the IN 51 determineds that milk should be refrigerated and stored at the farm what increased the importance of this group of microorganisms. In this work, psychrotrophic bacteria were isolated from 20 communitarian bulk tanks and 23 individual bulk tanks from dairy farms located at Zona da Mata region of Minas Gerais State and from southeastern Rio de Janeiro. Selected milk dilutions were plated on standard agar and after incubation for 10 days at 7oC, five colonies were isolated, firstly using nutrient agar and after using McConkey agar for 24 hours at 21oC. The isolates were identified by morphology, Gram stain method, catalase production, fermentative/oxidative metabolism and by API 20E, API 20NE, API Staph, API Coryne or API 50 CH (BioMerieux). In order to ensure reproductibility, API was repeated for 50% of the isolates. Species identification was considered when APILAB indexes reached 75% or higher. 309 strains were isolated, 250 Gram negative and 59 Gram positive. 250 Gram negative isolates were identified as: Acinetobacter spp. (39), Aeromonas spp. (07), A. Hydrophila (16), A. sobria (1), A. caviae (1), Alcaligenes feacalis (1), Burkholderia cepacia (12), Chryseomonas luteola (3), Enterobacter sp. (1), Ewingella americana(6), Hafnia alvei (7), Klebsiella sp. (1), Klebsiella oxytoca (10), Yersinia spp. (2), Methylobacterium mesophilicum (1), Moraxella spp. (4), Pantoea spp. (16), Pasteurella sp. (1), Pseudomonas spp. (10), P. fluorescens (94), P. putida (3), Serratia spp. (3), Sphigomonas paucomobilis (1). Five isolates kept unidentified. Pseudomonas was the predominant bacteria found (43%) and P. fluorescens the predominant species (37.6%), in accordance with previous reports. Qualitative analysis of proteolytic and lipolytic activity was based on halo formation using caseinate agar and tributirina agar during 72 hours at 21oC and during 10 days at 4°C, 10oC and 7°C. Among 250 Gram negative bacteria found, 104 were identified as Pseudomonas spp. and 60,57% of this group showed proteolytic and lipolytic acitivities over all four studied temperatures. 20% of Acinetobacter, Aeromonas, Alcaligenes, Burkholderia, Chryseomonas, Methylobacterium, Moraxella presented only lipolytic activity. Some isolates presented enzymatic activity in one or more studied temperatures. Among Gram positive bacteria, 30.51% were proteolytic and lipolytic at 10oC, 8.47% were proteolytic at 7oC, 10oC, and 21oC, 8.47% were proteolytic at all studied temperatures (4oC, 7oC, 10oC and 21oC) and 3.38% were proteolytic only at 21oC. At 4oC, only one isolate showed proteolytic activity and six isolates were lipolytic. In relation to Gram negative microorganisms, 4% were proteolytic and lipolytic at 7oC, 10oC and 21oC, 10% were proteolytic at 10oC and 4.4% were lipolytic at 4oC, 7oC, 10oC and 21oC, while 6.4% of all isolates were proteolytic and lipolytic at 10oC and 21oC as well as lipolytic at 4oC and 7oC. These findings are in accordance with previous researches that pointed out Pseudomonas as the predominant psycrotrophic flora in stored refrigerated raw milk

Influência da temperatura sobre fisiologia e crescimento de Xanthomonas citri subsp. citri

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Caracterização fenotípica e genotípica de bactérias recuperadas de implante ortopédico de conjunto placa-parafuso e parafuso de aço inoxidável austenítico após remoção cirúrgica

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação do potencial probiótico de bactérias acidoláticas produtoras de substância antimicrobiana isoladas de mussarela de búfala

Pós-graduação em Microbiologia - IBILCE

Bactérias produtoras de biossurfactantes: isolamento, produção, caracterização e comportamento num sistema modelo

Pós-graduação em Engenharia e Ciência de Alimentos - IBILCE

Efeito da terapia fotodinâmica antimicrobiana sobre culturas planctônicas e biofilmes de bactérias e fungos utilizando curcumina e cloro-alumínio ftalocianina veiculada por nanoemulsões

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ocorrência de Escherichia coli O157:H7 e O26 sorbitol negativas em matadouro frigorífico de bovino e suscetibilidade a antimicrobianos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Expressão de Pentraxina 3 em membranas corioamnióticas de gestações prétermo complicadas por rotura prematura de membranas pré-termo ou trabalho de parto prematuro

Pentraxina 3 (PTX3) é um componente essencial da resposta imune inata e age como receptores de reconhecimento de padrões (PPRs) reconhecendo produtos microbianos, opsonizando fungos, bactérias Gram positivas e negativas, além de apresentar a capacidade de ativar o sistema complemento. Poucos estudos na literatura mundial têm investigado a expressão de PTX3 nas complicações gestacionais associadas à invasão microbiana da cavidade amniótica e resultados conflitantes têm sido descritos. Avaliar a expressão gênica e protéica de PTX3 em membranas corioamnióticas de gestações pré-termo complicadas por Trabalho de Parto Prematuro (TPP) e bolsa íntegra ou Rotura Prematura de Membranas Pré-Termo (RPM-PT), na presença de corioamnionite histológica (CA). Foram incluídas no estudo 30 gestantes com TPP, sendo 15 na ausência e 15 na presença de corioamnionite histológica e 30 gestantes com RPM-PT, sendo 15 na ausência e 15 na presença de corioamnionite histológica. Cortes parafinizados foram encaminhados à análise histopatológica para confirmação de corioamnionite histológica. Outros fragmentos de 1cm2 das membranas foram submetidos à extração de RNA total. Após a extração do RNA, as amostras com concentração entre 0,02 e 0,2g/ L de RNA foram submetidas à obtenção de cDNA para posterior utilização na quantificação da expressão gênica de PTX3 pela técnica da PCR em tempo real empregando-se o Sistema TaqMan® Gene Expression Assays. Fragmentos das mesmas amostras incluídas no estudo foram utilizados para verificar a expressão da proteína PTX-3 através da técnica de Western Blotting. Dentre as 60 membranas corioamnióticas incluídas no estudo, 56 (93,3%) expressaram PTX-3. Não houve diferença estatisticamente significativa na expressão de mRNA de PTX-3 (p=0,137) entre os grupos: RPM-PT na presença de CA (Md: 0.355; 0.11-1.03), RPM-PT na...(Resumo completo, clicar acesso eletrônico abaixo)

Estudo das interações de peptídeos antimicrobianos e sistemas miméticos de membrana por dinâmica molecular

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Caracterização de segurança e tecnológica de bactérias acidoláticas termofílicas autóctones e aplicação em queijo parmesão

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Obtenção de consórcio de bactérias fotoheterotróficas geradoras de H2 visando sua produção em reator anaeróbio em batelada

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Identificação taxonômica e análise conexionista de bactérias heterotróficas e fatores abióticos de ambientes lóticos sob influência de efluentes do refino no petróleo

This study used a multi-analytical approach based on traditional microbiological methods for cultivation and isolation of heterotrophic bacteria in the laboratory associated with the molecular identification of the isolates and physicochemical analysis of environmental samples. The model chosen for data integration was supported by knowledge from computational neuroscience, and composed by three modules: (i) microbiological parameters, contemplating taxonomic data obtained from the partial sequencing of the 16S rRNA gene from 80 colonies of heterotrophic bacteria isolated by plating method in PCA media. For bacterial colonies isolation were used water samples from Atibaia and Jaguarí rivers collected at the site of water captation for use in effluent treatment, upstream from the entrance of treated effluent from the Paulínia refinery (REPLAN/Petrobras) located in the Paulínia-SP municipality, from the output of the biological treatment plant with stabilization pond and from the raw refinery wastewater; (ii) chemical parameters, ending measures of dissolved oxygen (DO), chemical oxygen demand (COD), biochemical oxygen demand (BOD), chloride, acidity CaCO3, alkalinity, ammonia, nitrite, nitrate, dissolved ions, sulfides, oils and greases; and (iii) physical parameters, comprising the pH determination, conductivity, temperature, transparency, settleable solids, suspended and soluble solids, volatile material, remaining fixing material (RFM), apparent color and turbidity. The results revealed interesting theoretical relationships involving two families of bacteria (Carnobacteriaceae and Aeromonadaceae). Carnobacteriaceae revealed positive theoretical relationships with COD, BOD, nitrate, chloride, temperature, conductivity and apparent color and negative theoretical relationships with the OD. Positive theoretical relationships were shown between Aeromonadaceae and OD and nitrate, while this bacterial family showed negative theoretical...

Identificação taxonômica e análise conexionista de bactérias heterotróficas e fatores abióticos de ambientes lóticos sob influência de efluentes do refino no petróleo

This study used a multi-analytical approach based on traditional microbiological methods for cultivation and isolation of heterotrophic bacteria in the laboratory associated with the molecular identification of the isolates and physicochemical analysis of environmental samples. The model chosen for data integration was supported by knowledge from computational neuroscience, and composed by three modules: (i) microbiological parameters, contemplating taxonomic data obtained from the partial sequencing of the 16S rRNA gene from 80 colonies of heterotrophic bacteria isolated by plating method in PCA media. For bacterial colonies isolation were used water samples from Atibaia and Jaguarí rivers collected at the site of water captation for use in effluent treatment, upstream from the entrance of treated effluent from the Paulínia refinery (REPLAN/Petrobras) located in the Paulínia-SP municipality, from the output of the biological treatment plant with stabilization pond and from the raw refinery wastewater; (ii) chemical parameters, ending measures of dissolved oxygen (DO), chemical oxygen demand (COD), biochemical oxygen demand (BOD), chloride, acidity CaCO3, alkalinity, ammonia, nitrite, nitrate, dissolved ions, sulfides, oils and greases; and (iii) physical parameters, comprising the pH determination, conductivity, temperature, transparency, settleable solids, suspended and soluble solids, volatile material, remaining fixing material (RFM), apparent color and turbidity. The results revealed interesting theoretical relationships involving two families of bacteria (Carnobacteriaceae and Aeromonadaceae). Carnobacteriaceae revealed positive theoretical relationships with COD, BOD, nitrate, chloride, temperature, conductivity and apparent color and negative theoretical relationships with the OD. Positive theoretical relationships were shown between Aeromonadaceae and OD and nitrate, while this bacterial family showed negative theoretical...

Avaliação do potencial de formigas (Hymenoptera: formicidae) como vetores mecânicos de bactérias do gêneroSstaphylococcus no ambiente hospitalar.

In a hospital environment, these bacteria can be spread by insects such as ants, which are characterized by high adaptability to the urban environment. Staphylococcus is a leading cause of hospital infection. In Europe, Latin America, USA and Canada, the group of coagulase negative staphylococci (CoNS) is the second leading cause of these infections, according to SENTRY (antimicrobial surveillance program- EUA). In this study, we investigated the potential of ants (Hymenoptera: Formicidae) as vehicle mechanics of Staphylococcus bacteria in a public hospital, in Natal-RN. The ants were collected, day and night, from June 2007 to may 2008, in the following sectors: hospitals, laundry, kitchen, blood bank. The ants were identified according to the identification key of Bolton, 1997. For the analysis of staphylococci, the ants were incubated in broth Tryptic Soy Broth (TSB) for 24 hours at 35 º C and then incubated on Mannitol Salt Agar. The typical colonies of staphylococci incubated for 24 hours at 35 ° C in Tryptic Soy Agar for the characterization tests (Gram stain, catalase, susceptibility to bacitracin and free coagulase). The identification of CoNS was performed through biochemical tests: susceptibility to novobiocin, growth under anaerobic conditions, presence of urease, the ornithine decarboxylation and acid production from the sugars mannose, maltose, trehalose, mannitol and xylose. The antimicrobial susceptibility examined by disk-diffusion technique. The technique of Polymerase Chain Reaction was used to confirm the presence of mecA gene and the ability to produce biofilm was verified by testing in vitro using polystyrene inert surface, in samples of resistant staphylococci. Among 440 ants, 85 (19.1%) were carrying coagulase-negative staphylococci (CoNS) of the species Staphylococcus saprophyticus (17), Staphylococcus epidermidis (15), Staphylococcus xylosus (13), Staphylococcus hominis hominis (10), Staphylococcus lugdunensis (10), Staphylococcus warneri (6), Staphylococcus cohnii urealyticum (5), Staphylococcus haemolyticus (3), Staphylococcus simulans (3), Staphylococcus cohnii cohnii (2), and Staphylococcus capitis (1). No Staphylococcus aureus was found. Among the isolates, 30.58% showed resistance to erythromycin. Two samples of CoNS (2.35%), obtained from the ant Tapinoma melanocephalum collected in the post-surgical female ward, S. Hominis hominis and S. lugdunensis harbored the mecA gene and were resistant to multiple antibiotics, and the specie S. hominis hominis even showed to be a biofilm producer. This study proves that ants act as carriers of multidrug-resistant coagulase-negative Staphylococci and biofilm producers and points to the risk of the spreading of pathogenic microorganisms by this insect in the hospital environment