In vitro characterization of virulence factors among species of the Candida parapsilosis complex

Abstract INTRODUCTION: Candida parapsilosis complex species differ from each other with regard to their prevalence and virulence. METHODS: The hydrolytic enzyme activity, biofilm production, and adhesion to epithelial cells were analyzed in 87 C. parapsilosis complex strains. RESULTS: Among the studied isolates, 97.7%, 63.2%, and 82.8% exhibited very strong proteinase, esterase, and hemolysin activity, respectively. All the C. parapsilosis complex isolates produced biofilms and presented an average adherence of 96.0 yeasts/100 epithelial cells. CONCLUSIONS: Our results show that Candida parapsilosis complex isolates showed different levels of enzyme activity, biofilm production, and adhesion to epithelial cells.

Evaluation of hollocelulase production by Lentinula edodes (Berk.) Pegler during the submerged fermentation growth using RSM / Avaliação da produção de enzimas holocelulolíticas por Lentinula edodes (Berk.) Pegler durante o crescimento em fermentação submersa usando MSR

The cellulase proteins have a great importance in the enzymatic hydrolysis of woody biomass. Despite of costs being a major concern, it has been a stimulus to study basidiomycetes biochemical properties which degrade lignocellulosic material and have prompted the processes' study for obtaining cellulolytic enzymes in fungi. The objective of this research was to evaluate the effects of the initial nitrogen content on (ammonium sulfate) and on sugar cane bagasse, which hereby, acts as an inducer of hydrolytic enzymes to produce cellulases and xylanases, using three Lentinula edodes (Berk.) Pegler strains as a transformation agent. A factorial design with 22 replications in the central point was conducted, varying concentrations of ammonium sulfate and sugar cane bagasse. The submerged cultures carried out in synthetic culture medium and incubated at 25°C for 7 days on an orbital shaker at 150 rpm. The total protein and cellulase activity as endoglucanase, exoglucanase and β-glucosidase and the xylanase was also determined. The results showed that the production of hydrolytic enzymes was stimulated by the presence of high concentrations of sugar cane bagasse (30g/L), characterizing it as an inducer due to the demonstrated proportional relationship. Thus, ammonium sulfate acted as a reducing agent in the synthesis of enzymes, being the low concentrations (0.1g/L) indicated for the enzyme production system under study. Among the studied strains, the EF52 showed higher activity for xylanase, endoglucanases, β-glucosidase and also protein.

As celulases são proteínas de grande importância na hidrólise enzimática de biomassa florestal. No entanto, seu custo elevado tem estimulado o estudo de processos de obtenção de enzimas celulolíticas por fungos filamentosos, tais como os basidiomicetos que apresentam propriedades bioquímicas para degradação de material lignocelulósico. O objetivo deste trabalho foi avaliar os efeitos do teor inicial de nitrogênio (sulfato de amônia) e de um indutor de enzimas hidrolíticas (bagaço de cana de açúcar) na produção de xilanases e celulases utilizando três isolados de Lentinula edodes (Berk.) Pegler como agente de transformação. Foi realizado um planejamento fatorial 22 com repetição no ponto central, variando as concentrações de sulfato de amônia e bagaço de cana de açúcar. O cultivo submerso realizado em meio de cultivo sintético e incubado a 25°C por 7 dias em agitador orbital a 150 rpm. Foram determinados o teor de proteínas totais e a atividade de celulase como: endoglucanase, exoglucanase e β-glucosidase e ainda xilanase. Os resultados demonstraram que a produção das enzimas hidrolíticas foi estimulada pela presença de alta concentração de bagaço de cana (30g/L), caracterizando-o como agente indutor devido à relação de proporcionalidade demonstrada. Por sua vez, o sulfato de amônio atuou como redutor da síntese de enzimas, sendo as baixas concentrações (0,1g/L) indicadas para o sistema de produção das enzimas em estudo. Quanto às linhagens, a EF52 mostrou maior atividade para xilanase, endoglucanases, β-glucosidase e proteínas.

Pathogenic and enzyme activities of the entomopathogenic fungus Tolypocladium cylindrosporum (Ascomycota: Hypocreales) from Tierra del Fuego, Argentina

Tolypocladium cylindrosporum is an entomopathogenic fungi that has been studied as a biological control agent against insects of several orders. The fungus has been isolated from the soil as well as from insects of the orders Coleoptera, Lepidoptera, Diptera and Hymenoptera. In this study, we analyzed the ability of a strain of T. cylindrosporum, isolated from soil samples taken in Tierra del Fuego, Argentina, to produce hydrolytic enzymes, and to study the relationship of those activities to the fungus pathogenicity against pest aphids. We have made the traditional and molecular characterization of this strain of T. cylindrosporum. The expression of hydrolase activity in the fungal strain was estimated at three incubation temperatures (4ºC, 12ºC and 24ºC), on different agar media supplemented with the following specific substrates: chitin azure, Tween ® 20, casein, and urea for chitinase, lipase, protease, and urease activity, respectively. The hydrolytic-enzyme activity was estimated qualitatively according to the presence of a halo of clarification through hydrolase action, besides was expressed semi-quantitatively as the ratio between the hydrolytic-halo and colony diameters. The pathogenicity of the fungus was tested on adults of the aphid Rhopalosiphum padi at three temperatures of incubation (4ºC, 12ºC and 24ºC). The suspension was adjusted to a concentration of 1x10 7 conidia/ml. In pathogenicity assays at seven days post-inoculation, the fungus caused the mortality of adults of Ropalosiphum padi at different temperatures also showed a broad ability to grow on several agar-culture media, supplemented with different carbon sources at the three incubation temperatures tested. Although, the growth was greater with higher incubation temperatures (with maximum levels at 24°C), the fungus reached similar colony diameters after 15 days of incubation on the medium supplemented with Tween® 20 at the lower two incubation temperatures of 4°C or 12°C. In accordance with the results on colony diameters, the fungus revealed an ability to degrade casein, chitin derivatives, Tween® 20, and urea as evidenced by the appearance of a halo around the fungal colony. Because of its origin and temperature tolerance, this Argentine strain has great potential for use as a biocontrol agent for insect pest control in cold and temperate environments. Rev. Biol. Trop. 60 (2): 833-841. Epub 2012 June 01.

El hongo entomopatógeno Tolypocladium cylindrosporum ha sido estudiado como un agente de control biológico contra insectos de varios órdenes. Esta especie fue aislada del suelo, así como de insectos de los órdenes Coleoptera, Lepidoptera, Diptera e Hymenoptera. En el presente trabajo hemos analizado la capacidad de una cepa de T. cylindrosporum (LPSC Nº1065) aislada del suelo en Tierra del Fuego, Argentina, para producir enzimas hidrolíticas y determinar la relación de esta actividad con la patogenicidad del hongo para combatir la plaga de los áfidos en diferentes temperaturas (4º, 12º y 24ºC). En los ensayos de patogenicidad, siete días posteriores a la inoculación, se registró mortalidad en los adultos del áfido Ropalosiphum padi a diferentes temperaturas y también se demostró una amplia capacidad de crecer en varios medios de cultivos complementados con diferentes fuentes de carbono bajo las tres temperaturas de incubación ensayadas. Debido a su origen y a la tolerancia que tiene a bajas temperaturas esta cepa, presenta un gran potencial para su uso como agente de control biológico para las plagas de insectos de ambientes fríos y templados.

Produção de fatores de virulência in vitro por espécies patogênicas do gênero Candida / Production of virulence factors in vitro by pathogenic species of the genus Candida

Avaliou-se, in vitro, a capacidade de crescimento em 39ºC e 42ºC, a produção de enzimas hidrolíticas e a atividade hemolítica de 21 cepas clínicas e de referência de sete espécies de Candida spp, Candida dubliniensis e Candida krusei demonstraram menor potencial de virulência e Candida albicans maior.

The growth capacity at 39ºC and 42ºC, production of hydrolytic enzymes and hemolytic activity of 21 clinical and reference strains of seven species of Candida spp were evaluated in vitro.Candida dubliniensis and Candida krusei demonstrated lower virulence potential and Candida albicans higher potential.

Diversity and effectiveness of tropical mangrove soil microflora on the degradation of polythene carry bags

The diversity and load of heterotrophic bacteria and fungi associated with the mangrove soil from Suva, Fiji Islands, was determined by using the plate count method. The ability of the bacterial isolates to produce various hydrolytic enzymes such as amylase, gelatinase and lipase were determined using the plate assay. The heterotrophic bacterial load was considerably higher than the fungal load. There was a predominance of the gram positive genus, Bacillus. Other genera encountered included Staphylococcus, Micrococcus, Listeria and Vibrio. Their effectiveness on the degradation of commercial polythene carry bags made of high density polyethylene (HDPE) and low density polyethylene (LDPE) was studied over a period of eight weeks in the laboratory. Biodegradation was measured in terms of mean weight loss, which was nearly 5 % after a period of eight weeks. There was a significant increase in the bacterial load of the soil attached to class 2 (HDPE) polythene. After eight weeks of submergence in mangrove soil, soil attached to class 1 and class 3 polythene mostly had Bacillus (Staphylococcus predominated in class 2 polythene). While most of the isolates were capable of producing hydrolytic enzymes such as amylase and gelatinase, lipolytic activity was low. Class 2 HDPE suffered the greatest biodegradation. Rev. Biol. Trop. 55 (3-4): 777-786. Epub 2007 December, 28.

Se determinó la diversidad y la carga de bacterias heterotróficas, así como los hongos asociados al suelo del manglar de Suva, Islas Fiji, utilizando el método de conteo de placas, usado también para medir la capacidad de bacterias aisladas para producir enzimas hidrolíticas como amilasa, gelatinasa y lipasa. La carga bacteriana heterotrófica resultó ser considerablemente más alta que la carga funguicida. Hubo predominancia de bacterias "Gram-positivas" del género de Bacillus. Otros géneros encontrados fueron Staphylococcus, Micrococcus, Listeria y Vibrio. La eficacia de esta microflora en la degradación del polietileno comercial de bolsas hechas de polietileno de alta densidad (HDPE) y de baja densidad (LDPE) fue estudiada en el laboratorio por un periodo de ocho semanas. La biodegradación fue medida en términos de pérdida de peso, la cual indicó una disminución del 5 %. Después de ocho semanas en el suelo de un manglar, el polietileno clase 1 y clase 3 contenía fundamentalmente Bacillus, pero en el polietileno clase 2 predominó el género Staphylococcus. Mientras que la mayoría de bacterias aisladas fueron capaces de producir enzimas hidrolíticas como la amilasa y la gelatinasa, la actividad lipolítica fue muy baja. La clase 2 (HDPE) experimentó la mayor biodegradación.

Regulation and overexpression of the bacillus subtilis -manganese operon

Some of the regulatory features of the Bacillus subtilis beta- mannanase operon [man A] have been investigated. The results proved that man A is extremely sensitive to cell growth state as a modulator, glucose, as a catabolite repressor and galactomannan as an inducer. It seems likely that catabolite repression of man A is mediated by two routes, one of which is commonly responsive to readily metabolizable carbon sources while the other is specific to glucose. At optimized physiological condition, the maximum enzyme yield [34 U/ml] was achieved within 4 hours. It was also demonstrated that mutated sporulation genes may block spore formation without affecting enzyme production. This would allow an extended enzyme production phase especially in continuous culture