Capacidade de invasão, persistência intracelular e atividade citotóxica de cepas de Aeromonas spp. em modelo celular in vitro e ex vivo

As Aeromonas são consideradas patógenos em potenciais para o homem e animais e estão amplamente distribuídas no ambiente sendo a água e os alimentos importantes veículos de transmissão. Muitos estudos têm demonstrado que a patologia causada pela infecção por Aeromonas é complexa e envolvem inúmeros fatores de virulência, dentre eles a aderência, invasão, enterotoxinas, hemolisinas, exoenzimas, sideróforos, flagelos, formação de biofilme e mecanismos de secreção. No presente estudo, analisamos os mecanismos de patogênese mediados por A. caviae e A. hydrophila, avaliando a participação desses microrganismos nos processos de adesão, invasão, persistência intracelular e citotoxidade celular. Foram utilizados ensaios quantitativos in vitro para testar associação, invasão e persistência intracelular em linhagens celulares HEp-2 e/ou T84. A interação de tecidos intestinais de coelho cultivados in vitro (IVOC) com três cepas de A. caviae originárias de fezes diarréicas também foi avaliada. Observamos que 10 (62,5%) das 16 cepas de Aeromonas spp. de diferentes origens, submetidas aos testes de invasão quantitativos foram capazes de invadir células HEp-2 e T84 em 6 horas de incubação. As cepas positivas nos testes de invasão foram submetidas ao teste quantitativo de persistência em células HEp-2 e sobreviveram no ambiente intracelular por 48 e/ou 72 horas sem multiplicação. A interação de três cepas de A. caviae com a mucosa intestinal de coelho ex vivo resultou em aderência, produção de muco e alterações como, intensa vacuolização e drástica desorganização estrutural que levaram a destruição das microvilosidades intestinais. Este estudo demonstrou que subconjuntos de cepas de A. caviae e A. hydrophila de diversas origens, foram capazes de invadir, persistir ou destruir linhagens celulares in vitro. Nosso estudo também evidenciou que cepas de A. caviae causaram expressivas alterações morfológicas que resultaram na destruição de epitélios intestinais de coelho ex vivo. Finalmente, nossos resultados contribuíram para reforçar o potencial patogênico de cepas de Aeromonas, em especial, as de origem vegetal e clínica.

Fungemia by yeasts in Brazil: occurrence and phenotypic study of strains isolated at the Public Hospital, Botucatu, São Paulo

Objective. - In this study strains of yeasts isolated from the blood of human patients were analyzed taxonomically, their virulence factors were determined and compared, and phenotypic markers were used to compare the samples with respect to phenotypic differences across the range of patients as well as between samples isotated from the same patient.Methods. - the study involved a total of 75 strains of yeast isolated from the blood of in-patients of the Public Hospital, Botucatu, S (a) over tildeo Paulo, Brazil, with a clinical profile of fungemia. The hospital wards with the largest number of fungemias were neonatal intensive care units (ICUs) (32%) followed by gastric surgery (13.4%) and pediatric wards (10.7%). After identification, the samples were analyzed for the production of phospholipase and proteinase enzymes, and biotyped according to their susceptibility to killer toxins.Results. - the most frequent species found was Candida albicans (38.7%) followed by C. parapsilosis (30.7%). In terms of enzyme production, 98.7% of the 75 samples of yeast presented a strongly positive activity for proteinase; however, 78.7% did not present any phospholipasic activity. Six different biotypes were identified, the most frequent being 511 and 888.Conclusion. In association with phenotypic methods, genetic analyses should also be made of the samples under study to help in the rational development of a wider range of preventive measures and better control of hospital-contracted infections. (c) 2005 Published by Elsevier SAS.

Estudo in vivo dos efeitos da terapia fotodinâmica, mediada pelo Photothazine® e luz led, sobre Cândida Albicans resistente a Fluconazol

Pós-graduação em Reabilitação Oral - FOAR

Streptococcus mutans-derived extracellular matrix in cariogenic oral biofilms

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

Atividade da fração enriquecida em fenólicos de Buchenavia tomentosa e de algumas substâncias isoladas antes e após encapsulação com beta-ciclodextrina em Candida albicans

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Activity and biomass of small benthic biota in the central Arctic Ocean

Sediment samples collected during the expedition "Arctic Ocean '96" with the Swedish ice-breaker ODEN were investigated to estimate for the first time heterotrophic activity and total microbial biomass (size range from bacteria to small metazoans) from the perennially ice-covered central Arctic Ocean. Benthic activities and biomass were evaluated analysing a series of biogenic sediment compounds (i.e. bacterial exoenzymes, total adenylates, DNA, phospholipids, particulate proteins). In contrast to the very time-consuming sorting, enumeration and weight determination, analyses of biochemical sediment parameters may represent a useful method for obtaining rapid information on the ecological situation in a given benthic system. Bacterial cell numbers and biomass were estimated for comparison with biochemically determined biomass data, to evaluate the contribution of the bacterial biomass to the total microbial biomass. It appeared that bacterial biomass made up only 8-31% (average of all stations = 20%) of the total microbial biomass, suggesting a large fraction of other small infaunal organisms within the sediment samples (most probably fungi, yeasts, protozoans such as flagellates, ciliates or amoebae, as well as a fraction of small metazoans). Activity and biomass values determined within this study were generally extremely low, and often even slightly lower than those given for other deep oceanic regions, thus characterizing the seafloor of the central Arctic Ocean as a "benthic desert". Nevertheless, some clear trends in the data could be found, e.g. generally sharply decreasing values within the sediment column, a vague tendency for declining values with increasing water depth of sampling stations, and also differences between various Arctic deep-sea regions.

Pigments of sediment cores of the Northwest European Continental Margin

In the context of the European OMEX Programme this investigation focused on gradients in the biomass and activity of the small benthic size spectrum along a transect across the Goban Spur from the outer Celtic Sea into Porcupine Abyssal Plain. The effects of food pulses (seasonal, episodic) on this part of the benthic size spectrum were investigated. Sediments sampled during eight expeditions at different seasons covering a range from 200 m to 4800 m water depth were assayed with biochemical bulk measurements: determinations of chloroplastic pigment equivalents (CPE), the sum of chlorophyll a and its breakdown products, provide information concerning the input of phytodetrital matter to the seafloor; phospholipids were analyzed to estimate the total biomass of small benthic organisms (including bacteria, fungi, flagellata, protozoa and small metazoan meiofauna). A new term 'small size class biomass' (SSCB) is introduced for the biomass of the smallest size classes of sediment-inhabiting organisms; the reduction of fluorescein-di-acetate (FDA) was determined to evaluate the potential activity of ester-cleaving bacterial exoenzymes in the sediment samples. At all stations benthic biomass was predominantly composed of the small size spectrum (90% on the shelf; 97-98% in the bathyal and abyssal parts of the transect). Small size class biomass (integrated over a 10 cm sediment column) ranged from 8 g C/m**2 on the shelf to 2.1 g C/m**2 on the adjacent Porcupine Abyssal Plain, exponentially decreasing with increasing water depth. However, a correlation between water depth and SSCB, macrofauna biomass as well as metazoan meiofauna biomass exhibited a significantly flatter slope for the small size classes in comparison to the larger organisms. CPE values indicated a pronounced seasonal cycle on the shelf and upper slope with twin peaks of phytodetrital deposition in mid spring and late summer. The deeper stations seem to receive a single annual flux maximum in late summer. SSCB and heterotrophic activity are significantly correlated to the amount of sediment-bound pigments. Seasonality in pigment concentrations is clearly followed by SSCB and activity. In contrast to macro- and megafauna which integrate over larger periods (months/years), the small benthic size classes, namely bacteria and foraminifera, proved to be the most reactive potential of the benthic communities to any perturbations on short time scales (days/weeks). The small size classes, therefore, occupy a key role in early diagenetic processes.