Nutritional-requirements of Cryptococcus-laurentii in sugar-cane vinasse medium

The growth of the yeast Cryptococcus laurentii in complete broth for yeast and in liquid and semi-solid sugar cane vinasse media supplemented with several nitrogen and phosphorus sources was analysed in order to evaluate its potential utilization as biomass producer from vinasse. The trials were performed in a 1-liter fermentor, at 30-degrees-C, under magnetic agitation for 48 hours, and initial pH 5.0 for the liquid media. The parameters analysed were biomass, protein, and final pH. For the semi-solid media, agitated flasks under rotational agitation for 24 hours, at 30-degrees-C, and initial pH 5.0 were used. The results obtained showed that the yeast did not grow well in liquid sugar cane vinasse media, both supplemented or not, in comparison to the complete broth for yeast. The protein content was also lower in liquid sugar cane vinasse media. The medium pH did not alter markedly during the cultivation. The best results were obtained in semi-solid sugar cane vinasse media with supplementation, probably due to the known polysaccharide production by this species, which allows a better survival to solid substrates.

Avaliação audiomética em pacientes tratados de meningite por Cryptococcus gattii

Este estudo mostra a avaliação audiométrica, composta de audiometria tonal limiar e imitanciometria, em pacientes tratados de meningite por Cryptococcus gattii. Trata-se de um estudo tipo relato de casos. Foram avaliados seis pacientes com infecção comprovada do sistema nervoso central pelo Cryptococcus gattii, internados no Hospital Universitário João de Barros Barreto, entre janeiro de 2000 a novembro de 2007, para tratamento antifúngico. A idade dos pacientes analisados variou entre 12 e 37 anos. Três eram homens e três mulheres. Todos procedentes de municípios vizinhos, não sendo ninguém da capital Belém. As principais manifestações clínicas foram cefaléia e vômitos, presentes nas histórias de todos os pacientes, seguidas de febre e rigidez de nuca, cada uma delas ausentes em apenas um paciente, não coincidentes. O tempo de doença até o diagnóstico variou de 3 a 8 semanas. Dos três homens avaliados, dois apresentaram perda auditiva. Num deles a perda foi moderada e unilateral e no outro foi assimétrica e bilateral. Das três mulheres, apenas uma apresentou alteração auditiva. Nessa paciente houve apenas um rebaixamento leve numa freqüência isolada, de 4.000 Hz. Descreve-se achados audiométricos em relato de casos de pacientes tratados de meningite por Cryptococcus gattii.

Perfil clínico-epidemiológico da Criptococose em pacientes HIV positivos atendidos em uma unidade de referência em Belém do Pará

A criptococose em residentes no estado do Pará apresenta-se como endemia grave, ocorrendo nas diferentes mesorregiões do estado e na área metropolitana de Belém. Ocorre causando neurocriptococose, com letalidade acima de 40%. Para tentar elucidar o comportamento clínico e epidemiológico da criptococose em pacientes HIV positivos, realizamos um estudo retrospectivo, analítico de série de casos, com revisão de prontuários dos últimos 05 anos (2006 –2010), com pacientes HIV positivos em um hospital de referência em Belém – Pará. A doença ainda é uma das causas mais freqüentes de óbitos em pacientes HIV positivos na região norte e deve-se ao C. neoformans o maior número de casos, contribuindo com 57 (60,6%) de um total de 94 casos. No que diz respeito à forma clínica, da criptococose, desenvolvida pelos pacientes HIV positivos, observou-se que o acometimento do sistema nervoso está presente em 46,8% dos pacientes e a disseminação da doença em 27,7% dos casos. A relação entre a causa do óbito e a presença do Cryptococcus, ocorre por um conjunto complexo de fatores que devem ser esclarecidos e correlacionados com o óbito dos pacientes e o mais importante, estabelecerem o quão forte é essa relação uma vez detectada.

Ancient Dispersal of the Human Fungal Pathogen Cryptococcus gattii from the Amazon Rainforest

Over the past two decades, several fungal outbreaks have occurred, including the high-profile 'Vancouver Island' and 'Pacific Northwest' outbreaks, caused by Cryptococcus gattii, which has affected hundreds of otherwise healthy humans and animals. Over the same time period, C. gattii was the cause of several additional case clusters at localities outside of the tropical and subtropical climate zones where the species normally occurs. In every case, the causative agent belongs to a previously rare genotype of C. gattii called AFLP6/VGII, but the origin of the outbreak clades remains enigmatic. Here we used phylogenetic and recombination analyses, based on AFLP and multiple MLST datasets, and coalescence gene genealogy to demonstrate that these outbreaks have arisen from a highly-recombining C. gattii population in the native rainforest of Northern Brazil. Thus the modern virulent C. gattii AFLP6/VGII outbreak lineages derived from mating events in South America and then dispersed to temperate regions where they cause serious infections in humans and animals.

Eugenia calycina Cambess extracts and their fractions: Their antimicrobial activity and the identification of major polar compounds using electrospray ionization FT-ICR mass spectrometry

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Immunoreactivity of synthetic peptides derived from proteins of Cryptococcus gattii

Aim: To determine the immunoreactivity of synthetic Cryptococcus-derived peptides. Materials & methods: A total of 63 B-cell epitopes from previously identified Cryptococcus gattii immunoreactive proteins were synthesized and evaluated as antigens in ELISAs. The peptides were first evaluated for their ability to react against sera from immunocompetent subjects carrying cryptococcal meningitis. Peptides that yielded high sensitivity and specificity in the first test were then retested with sera from individuals with other fungal pathologies for cross-reactivity determination. Results: Six of 63 synthetic peptides were recognized by antibodies in immunoassays, with a specificity of 100%, sensitivity of 78% and low cross-reactivity. Conclusion: We successfully determined the immunoreactivity of selected synthetic peptides of C. gattii derived proteins.

Triagem de substâncias antifúngicas com atividade inibitória de bombas de efluxo de isolados resistentes de Cryptococcus sp

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Análise do potencial de micro-organismos dos gêneros Lectthophora e Cryptococcus para a produção de biocumbustíveis

A busca de novas fontes de energia tem sido estimulada pela demanda crescente do mercado e o bioetanol é uma alternativa promissora para esta questão. A produção de etanol a partir de resíduos agroindustriais lignocelulósicos está sendo pesquisada em vários laboratórios do Brasil e do exterior. Os resíduos da agroindústria são formados, em grande parte, por fibra vegetal lignocelulósica, a qual é altamente resistente à degradação, dificultando a conversão dos componentes a monossacarídeos fermentescíveis. Desta forma, a prospecção de micro-organismos que viabilizem a conversão de material lignocelulósico em etanol tem aumentado consideravelmente. Sendo assim, o estudo de micro-organismos associados às formigas cortadeiras, os quais são adaptados ao uso da biomassa lignocelulolítica, pode ser promissor para a descoberta de enzimas adequadas à degradação eficiente da fibra vegetal, podendo significar um passo adiante na produção do etanol de segunda geração. Neste trabalho analisamos a ação hidrolítica e a liberação de açúcares fermentescíveis por fungos dimórficos do gênero Lecythophora e de leveduras do gênero Cryptococcus sobre o material lignocelulósico. A metodologia empregada consistiu na inoculação das linhagens em diferentes resíduos da agroindústria, como bagaço de cana-de-açúcar, casca de café, casca de arroz e sabugo de milho triturado, seguida da quantificação dos açúcares redutores liberados, bem como da produção de enzimas hidrolíticas extracelulares, como endoamilase, exoamilase, pectinase, endoglucanase, exoglucanase e xilanase. O gênero Lecythophora mostrou-se melhor produtor enzimático quando comparado com o gênero Cryptococcus, ambos em meio suplementado com bagaço de cana-de-açúcar. Ainda, foi realizado o inóculo de culturas mistas das melhores linhagens pertencentes ao gênero Lecythophora. A produção enzimática constatada indicou o potencial das linhagens...

Microbiota fúngica de passeriformes de cativeiros da Região Noroeste do Estado de São Paulo

Birds are hosts for a rich fungal microbiota which can act as potent pathogens for humans and other species of animals, causing thereby serious public health problems. The objective of this study was to evaluate the participation of birds kept in containers in the epidemiology of infectious diseases such as cryptococcosis and aspergillosis, thus verifying the maintenance and spread of pathogens in the environment. 36 samples of excretas of passeriformes were collected and were cultivated in Sabouraud Dextrose Agar 4% at room temperature and 37°C. The isolated fungal colonies were classified according to their morphological and staining characteristics. Subsequently, those in yeast form were peaked in Niger Agar, incubated at 30°C. In one sample showed growth of more than one type of colony and there was verified the presence of 25.0% of Penicillium spp., 19.4% of Trichosporon spp., 13.9% of C. gattii, 11.1% of C. neoformans, 11.1% of Candida spp., 8.3% of Rhizomucor spp., 8.3% of Aspergillus spp., 2.8% of Nigrospora spp. and 2,8% of Geotrichum spp. It can be conluded by the expost that birds shed continuously pathogenic microorganisms in their feces acting in definitive form in the infectious diseases ecoepidemiology.

Cryptococcus haglerorum, sp nov., an anamorphic basidiomycetous yeast isolated from nests of the leaf-cutting ant Atta sexdens

A yeast strain (CBS 8902) was isolated from the nest of a leaf-cutting ant and was shown to be related to Cryptococcus humicola. Sequencing of the D1/D2 region of the 26S ribosomal DNA and physiological characterization revealed a separate taxonomic position. A novel species named Cryptococcus haglerorum is proposed to accommodate strain CBS 8902 that assimilates n-hexadecane and several benzene compounds. Physiological characteristics distinguishing the novel species from some other members of the C. humicola complex are presented. The phylogenetic relationship of these strains to species of the genus Trichosporon Behrend is discussed.

Virulence of Cryptococcus sp. biofilms in vitro and in vivo using Galleria mellonella as an alternative model.

2016

A multi-criteria analysis approach for ranking and selection of microorganisms for the production of oils for biodiesel production

Oleaginous microorganisms have potential to be used to produce oils as alternative feedstock for biodiesel production. Microalgae (Chlorella protothecoides and Chlorella zofingiensis), yeasts (Cryptococcus albidus and Rhodotorula mucilaginosa), and fungi (Aspergillus oryzae and Mucor plumbeus) were investigated for their ability to produce oil from glucose, xylose and glycerol. Multi-criteria analysis (MCA) using analytic hierarchy process (AHP) and preference ranking organization method for the enrichment of evaluations (PROMETHEE) with graphical analysis for interactive aid (GAIA), was used to rank and select the preferred microorganisms for oil production for biodiesel application. This was based on a number of criteria viz., oil concentration, content, production rate and yield, substrate consumption rate, fatty acids composition, biomass harvesting and nutrient costs. PROMETHEE selected A. oryzae, M. plumbeus and R. mucilaginosa as the most prospective species for oil production. However, further analysis by GAIA Webs identified A. oryzae and M. plumbeus as the best performing microorganisms.

Microbial oil production from palm oil empty fruit bunch hydrolysates

This study explores the potential use of empty fruit bunch (EFB) residues from palm oil processing residues, as an alternative feedstock for microbial oil production. EFB is a readily available, lignocellulosic biomass that provides cheaper substrates for oil production in comparison to the use of pure sugars. In this study, potential oleaginous microorganisms were selected based on a multi-criteria analysis (MCA) framework which utilised Analytical Hierarchy Process (AHP) with Preference Ranking Organization Method for Enrichment Evaluation (PROMETHEE) aided by Geometrical Analysis for Interactive Aid (GAIA). The MCA framework was used to evaluate several strains of microalgae (Chlorella protothecoides and Chlorella zofingiensis), yeasts (Cryptococcus albidus and Rhodotorula mucilaginosa) and fungi (Aspergillus oryzae and Mucor plumbeus) on glucose, xylose and glycerol. Based on the results of PROMETHEE rankings and GAIA plane, fungal strains A. oryzae and M. plumbeus and yeast strain R. mucilaginosa showed great promise for oil production from lignocellulosic hydrolysates. The study further cultivated A. oryzae, M. plumbeus and R. mucilaginosa on EFB hydrolysates for oil production. EFB was pretreated with dilute sulfuric acid, followed by enzymatic saccharification of solid residue. Hydrolysates tested in this study are detoxified liquid hydrolysates (LH) and enzymatic hydrolysate (EH).

Gut-inhabiting yeasts from the gut of Australian passalid beetles

Passalidae (Polyphaga, Coleoptera) is a family of beetles with approximately 960 species that are distributed worldwide. Preliminary studies that characterized ascomycete and basidiomycete yeasts in the gut of these wood-eating beetles from the USA, Guatemala, and Thailand, demonstrated associations between certain yeast taxa and passalids. We extended the study to include yeasts and beetles from tropical forests near Cairns and Brisbane, Queensland, Australia. We isolated more than 1000 yeast strains from about 150 beetles belonging to 10 species. LSU and ITS rRNA markers were used to identify a subset of 250 yeast strains, which revealed that the gut of Australian passalids contained undescribed ascomycetes in the Debaryomyces, Dipodascus, Kazachstania, Ogataea, Scheffersomyces, Sugiyamaella, Spathaspora, Torulaspora, and Zygowilliopsis clades, as well as basidiomycetes in the genera Cryptococcus and Trichosporon. A close relative of Candida subhashii (Spathaspora clade) and the xylose-fermenting yeast Scheffersomyces stipitis were the most common species isolated in Queensland. These results agree with those of previous studies that showed a common association of xylose-fermenting yeasts in the gut of lignicolous insects. Species and higher taxa of yeasts, however, vary between Queensland passalids and those previously collected in distant regions.