Candida parapsilosis complex water isolates from a haemodialysis unit: biofilm production and in vitro evaluation of the use of clinical antifungals

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

A nested-PCR assay for detection of Xylella fastidiosa in citrus plants and sharpshooter leafhoppers

Aims: Detection of Xylella fastidiosa in citrus plants and insect vectors.Methods and Results: Chelex 100 resin matrix was successfully standardized allowing a fast DNA extraction of X. fastidiosa. An amplicon of 500 bp was observed in samples of citrus leaf and citrus xylem extract, with and without symptoms of citrus variegated chlorosis, using PCR with a specific primer set indicating the presence of X. fastidiosa. The addition of insoluble acid-washed polyvinylpyrrolidone (PVPP) prior to DNA extraction of insect samples using Chelex 100 resin together with nested-PCR permitted the detection of X. fastidiosa within sharpshooter heads with great sensitivity. It was possible to detect up to two bacteria per reaction. From 250 sharpshooter samples comprising four species (Dilobopterus costalimai, Oncometopia facialis, Bucephalogonia xanthopis and Acrogonia sp.), 87 individuals showed positive results for X. fastidiosa in a nested-PCR assay.Conclusions: the use of Chelex 100 resin allowed a fast and efficient DNA extraction to be used in the detection of X. fastidiosa in citrus plants and insect vectors by PCR and nested-PCR assays, respectively.Significance and Impact of the study: the employment of efficient and sensitive methods to detect X. fastidiosa in citrus plants and insect vectors will greatly assist epidemiological studies.

Chemical and microbiological changes and aerobic stability of marandu grass silages after silo opening

Esta pesquisa foi realizada com os objetivos de caracterizar a microbiologia e avaliar a estabilidade aeróbia de silagens de capim-marandu contendo polpa cítrica peletizada (PCP). A forragem colhida foi submetida aos seguintes tratamentos: silagem de capim-marandu; silagem de capim-marandu + 5% PCP e silagem do capim-marandu + 10% de PCP com base na matéria natural. As alterações químicas e microbiológicas foram feitas aos 0, 2, 4 e 6 dias após a abertura dos silos (tambores de metal com 80 cm de altura e 50 cm de diâmetro). Na avaliação da estabilidade aeróbia por meio da alteração da temperatura, 3 kg de silagem foram colocados em caixas de isopor, que foram armazenadas em câmara climática. Nos dois ensaios realizados, utilizou-se delineamento inteiramente ao acaso com cinco repetições, em esquema de parcelas subdivididas, de modo que os níveis de PCP na silagem corresponderam às parcelas e o tempo, às subparcelas. As silagens sem polpa apresentaram desenvolvimento de bacilos e enterobactérias e aumento do pH no decorrer do desabastecimento dos silos. A presença de leveduras foi detectada nas silagens contendo o aditivo e aumentou do primeiro ao sexto dia de aeração. A digestibilidade in vitro da matéria seca (DIVMS) reduziu com o aumento dos tempos de aeração. A temperatura nas silagens com 10% de PCP aumentou durante a aerobiose. Com a abertura dos silos, as silagens apresentaram-se instáveis, seja pelo desenvolvimento de bactérias seja pelo desenvolvimento de leveduras, o que reduziu seu valor nutritivo.

Fermentative and microbiological profile of marandu-grass ensiled with citrus pulp pellets

Forragens com alta umidade e baixa concentração de carboidratos solúveis, como é o caso dos capins tropicais, podem conduzir a condições desfavoráveis para a produção de silagens de qualidade satisfatória. Esse estudo objetivou conhecer o perfil fermentativo e microbiológico de silagens de capim-Marandu [Brachiaria brizantha (Hochst ex. A. Rich) Stapf cv. Marandu], colhido com 58 dias de crescimento, aditivadas com polpa cítrica peletizada (PCP),. Foram utilizados silos experimentais de PVC adaptados com válvula do tipo Bunsen, tendo a silagem atingindo densidade de 900 kg m-3. Os tratamentos foram constituídos por três proporções de PCP (0, 50 e 100 g kg-1 em relação a matéria natural) e sete tempos de abertura após a ensilagem (1, 4, 7, 14, 21, 28 e 56 dias). A presença de PCP aumentou os teores de CHOs em 15 a 20%, reduziu o pH (5,3 para 4,2) e diminuiu as concentrações de N-NH3. Houve crescimento da população de enterobactérias somente durante o primeiro dia de fermentação (média 3 UFC g-1), pequeno desenvolvimento de clostrideos (média 0,5 UFC g-1) e dominância de bactérias homo em relação às heterofermentativas, para os três tratamentos estudados. A adição de polpa cítrica durante a ensilagem do capim-Marandu foi benéfica, podendo ser recomendada desde que haja benefício econômico na sua adoção.

Genotoxicity and antigenotoxicity assessment of shiitake (Lentinula edodes (Berkeley) Pegler) using the Comet assay

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

Evaluation of physicochemical, microbiological and sensorial characteristics of fermented milk beverages with buttermilk addition

This study evaluated three fermented milk beverages to which had been added sugar strawberry puree post-fermentation. The base was composed of 70% of milk, with whey and buttermilk in the concentrations of 30% and 0%, 15% and 15%, and 0% and 30%, respectively. The starter culture developed well with all formulations reaching pH 4.74.9 in 180 min of fermentation. Lactic acid bacteria in the products were above 8 log cfu/mL throughout the study. The beverages presented similar pH, acidity and viscosity. Buttermilk and whey can be interesting ingredients to be added into fermented milk beverages, because the consumers liked all the products equally, which had an average acceptance score exceeding liked moderately.

Evaluation of curcumin and cisplatin-induced DNA damage in PC12 cells by the alkaline comet assay

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Investigation of the Photodynamic Effects of Curcumin Against Candida albicans

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

MICROBIOLOGICAL CONTAMINATION of A HEMODIALYSIS CENTER WATER DISTRIBUTION SYSTEM

The microbiological monitoring of the water used for hemodialysis is extremely important, especially because of the debilitated immune system of patients suffering from chronic renal insufficiency. To investigate the occurrence and species diversity of bacteria in waters, water samples were collected monthly from a hemodialysis center in upstate São Paulo and tap water samples at the terminal sites of the distribution system was sampled repeatedly (22 times) at each of five points in the distribution system; a further 36 samples were taken from cannulae in 19 hemodialysis machines that were ready for the next patient, four samples from the reuse system and 13 from the water storage system. To identify bacteria, samples were filtered through 0.22 mu m-pore membranes; for mycobacteria, 0.45 mu m pores were used. Conventional microbiological and molecular methods were used in the analysis. Bacteria were isolated from the distribution system (128 isolates), kidney machine water (43) and reuse system (3). Among these isolates, 32 were Gram-positive rods, 120 Gram-negative rods, 20 Gram-positive cocci and 11 mycobacteria. We propose the continual monitoring of the water supplies in hemodialysis centers and the adoption of effective prophylactic measures that minimize the exposure of these immunodeficient patients to contaminated sources of water.

Evaluation of four molecular typing Methodologies as tools for determining taxonomy relations and for identifying. species among Yersinia isolates

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

In vivo assessment of DNA damage and protective effects of extracts from Miconia species using the comet assay and micronucleus test

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

Rapid detection of resistance to pyrazinamide in Mycobacterium tuberculosis using the resazurin microtitre assay

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Mutagenicity of New Lead Compounds to Treat Sickle Cell Disease Symptoms in a Salmonella/Microsome Assay

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Genotoxicity of Brosimum gaudichaudii measured by the Salmonella/microsome assay and chromosomal aberrations in CHO cells

The root bark of Brosimum gaudichaudii Trecul (Moraceae) is popularly used for treatment of vitiligo. In the present study the mutagenic activity of the aqueous and methanolic extract as well as of the n-butanolic fraction of this medicinal plant were evaluated using Salmonella typhimurium assays, TA100, TA98, TA102 and TA97a strains, while the clastogenic effect in Chinese hamster ovary (CHO) cells in the G(1)/S, S and G(2)/S phases of the cell cycle. The results showed mutagenic activity of the aqueous extract against TA102 in the presence of S9, and of methanolic extract, with and without metabolic activation. TA100 mutagenicity was only observed for the methanolic extract in the absence of S9. The n-butanolic fraction did not present mutagenic activity. In CHO cells only the methanolic extract induced a significant increase of chromosomal aberrations in the G(1)/S and S phases, whereas a decrease in the mitotic index was observed in the G(1)/S and G(2)/S phases. No clastogenicity was observed for the aqueous extract. The furocoumarins (psoralen and bergapten) presented in the extracts might contribute to the mutagenicity. The lower activity of the aqueous extract was probably due to the presence of smaller amount of furocoumarins compared to the methanolic extract. (C) 2002 Elsevier B.V. Ireland Ltd. All rights reserved.

p-iodophenol-enhanced luminol chemiluminescent assay applied to discrimination between acute lymphoblastic and minimally differentiated acute myeloid (FAB-M0) or acute megakaryoblastic (FAB-M7) leukemias

Introduction: In this report, we propose the application of the p-iodophenol-enhanced luminol chemiluminescent technique to the determination of peroxidase (myeloperoxidase and/or platelet peroxidase) activity in blasts of minimally differentiated acute myeloblastic leukemia (AML-M0) and acute megakaryoblastic leukemia (AML-M7).Methods: the frozen blast cells from 29 patients were thawed and submitted to the optimized protocol.Results: All cases of AML-M7 and AML-M0 exhibited integrated light emission greater than 73 (10(2) mV x s), which was the arbitrary cutoff point set for the discrimination between AML and acute lymphoblastic leukemia (ALL) (mean + 3 x s.d. of ALL samples, n = 10). In addition, five out of seven cases of AML-M0 showed results above the Cutoff point.Conclusion: This highly sensitive enhanced chemiluminescent technique may be applied to discriminate between ALL and AML-M7 or AML-M1 cases, and most AML-M0 cases. It is very simple, cheap and easy to perform compared to other procedures used to measure MPO activity in AML-leukemias including AML-M7 and AML-M0.