Cryptococcosis outbreak in psittacine birds in Brazil

An outbreak of cryptococcosis occurred in a breeding aviary in São Paulo, Brazil. Seven psittacine birds (of species Charmosyna papou, Lorius lory, Trichoglossus goldiei, Psittacula krameri and Psittacus erithacus) died of disseminated cryptococcosis. Incoordination, progressive paralysis and difficulty in flying were seen in five birds, whereas superficial lesions coincident with respiratory alterations were seen in two birds. Encapsulated yeasts suggestive of Cryptococcus sp. were seen in faecal smears stained with India ink in two cases. Histological examination of the birds showed cryptococcal cells in various tissues, including the beak, choana, sinus, lungs, air sacs, heart, liver, spleen, kidneys, intestines and central nervous system. High titres of cryptococcal antigen were observed in the serum of an affected bird. In this case, titres increased during treatment and the bird eventually died. Yeasts were isolated from the nasal mass, faeces and liver of one bird. Cryptococcus neoformans var. gattii serovar B was identified based on biochemical, physiological and serological tests. These strains were resistant (minimum inhibitory concentration 64 μ g/ml) to fluconazole. This is the first report of C. neoformans var. gattii occurring in psittacine birds in Brazil. © 2004 ISHAM.

Comparative study of disk diffusion and microdilution methods for evaluation of antifungal activity of natural compounds against medical yeasts Candida spp and Cryptococcus sp

Antifungal activity of natural products has been tested by adapting methods designed for synthetic drugs. In this study, two methods for the determination of antifungal activity of natural products, agar diffusion and broth microdilution, the CLSI reference methods for synthetic drugs, are compared and discussed. The microdilution method was more sensitive. The minimal inhibitory concentrations (MIC) of crude extracts, fractions and pure substances from different species of the plant families Piperaceae, Rubiaceae, Clusiaceae, Fabaceae and Lauraceae, from the Biota project, were determined. Antifungal activities against Candida albicans, C.krusei, C.parapsilosis and Cryptococcus neoformans were produced by several samples.

Cryptococcosis: epidemiology, fungal resistance, and new alternatives for treatment

Cryptococcosis is an important systemic mycosis and the third most prevalent disease in human immunodeficiency virus (HIV)-positive individuals. The incidence of cryptococcosis is high among the 25 million people with HIV/acquired immunodeficiency syndrome (AIDS), with recent estimates indicating that there are one million cases of cryptococcal meningitis globally per year in AIDS patients. In Cryptococcus neoformans, resistance to azoles may be associated with alterations in the target enzyme encoded by the gene ERG11, lanosterol 14α-demethylase. These alterations are obtained through mutations, or by overexpressing the gene encoding. In addition, C. gattii and C. neoformans present a heteroresistance phenotype, which may be related to increased virulence. Other species beyond C. neoformans and C. gattii, such as C. laurentii, have been diagnosed mainly in patients with immunosuppression. Infections of C. albidus have been isolated in cats and marine mammals. Recent evidence suggests that the majority of infections produced by this pathogen are associated with biofilm growth, which is also related with increased resistance to antifungal agents. Therefore, there is a great need to search for alternative antifungal agents for these fungi. The search for new molecules is currently occurring from nanoparticle drugs of plant peptide origin. This article presents a brief review of the literature regarding the epidemiology of cryptococcosis, as well as fungal resistance and new alternatives for treatment. © 2013 Springer-Verlag Berlin Heidelberg.

Sensibilidade à antifúngicos sintéticos e naturais anti-Cryptococcus neoformans determinados por citometria de fluxo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Perfil fenotípico e de expressão de proteínas de Cryptococcus neoformans após tratamento com substâncias obtidas da planta Pterogyne nitens

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Cryptococcus neoformans de fontes ambientais e biopropecção de extratos de plantas com atividade saneante

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

Caracterização morfológica e fenotípica de leveduras do gênero Cryptococcus com ênfase na pesquisa de Cryptococcus neoormans isoladas de amostras de água de mar e areia de três ambientes costeiros do estado de São Paulo: Canal de São Sebastião, Baixada Santista e Ubatuba

The yeast Cryptococcus neoformans is the etiologic agent of cryptococcosis, an infectious cosmopolitan disease that affects humans. Although rare, this disease is potentially fatal, especially for immunocompromised hosts. This pathogen is frequently isolated from excrements of pigeons and parrots, with many environmental sources such as birds, pigeon droppings, eucalyptus leaves, decaying trees, towers, churches and places of storage of grain (the port area). The isolation of this microorganism has been obtained also from the aquatic environment. The identification of environmental sources is needed to protect human health, especially susceptible populations such as immunocompromised. Therefore, this study investigated the presence of Cryptococcus neoformans in yeast isolates obtained from samples of sea water and sand from three regions of São Paulo: São Sebastião Channel, Santos and Ubatuba. Isolates were analyzed according to micro-and macroscopic characteristics and biochemical tests: microculture, urease, ink nankin, auxanograma, zymogram and phenol. We analyzed 199 isolates, 175 of which had features suggestive for Cryptococcus spp. in microculture. All these 175 isolates were sown in the Christensen urea middle to verify the production of urease and submitted to the technique nankin ink to visualize the capsule. Of these, only 24 were selected for the next test that was the auxanograma (assimilation of carbohydrate and nitrogen). Of the 24, 10 were tested in zymograms (fermented sugar), from which 5 were selected for the phenoloxidase test in medium containing dopamine. None of the 5 isolates tested had black or brown color characteristic of Cryptococcus neoformans. According to these tests, we arrived at 5 isolates identified to the genus Cryptococcus, but not the neoformans specie

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)

Composition and antifungal activity against Candida albicans, C. parapsilopisis, Candida krusei and Cryptococcus neoformans of essential oils from leaves of Piper and Peperomia species

This study was addressed to investigate the composition and antifungal activity of essential oils from leaves of Piperaceae species (Piper aduncum, Piper amalago, Piper cernuum, Piper diospyrifolium, Piper crassinervium, Piper gaudichaudianum, Piper solmsianum, Piper regnellii, Piper tuberculatum, Piper umbelata and Peperomia obtusifolia) against Candida albicans, C. parapsilosis, C. krusei and Cryptococcus neoformans. The essential oils from P. aduncum, P. gaudichaudianum and P. solmsianum showed the highest antifungal activity against Cryptococcus neoformans with the MIC of 62.5, 62.5 and 3.9 mg.mL-1, respectively. The oil from P. gaudichaudianum showed activity against C. krusei with MIC of 31.25 mg.mL-1.

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.

In Vitro Photodynamic Inactivation of Cryptococcus neoformans Melanized Cells with Chloroaluminum Phthalocyanine Nanoemulsion

The selection of fungi resistant to currently used fungicides and the emergence of new pathogenic species make the development of alternative fungus-control techniques highly desirable. Photodynamic antimicrobial chemotherapy (PACT) is a promising method which combines a nontoxic photosensitizer (PS) with visible light to cause selective killing of microbial cells. The development of PACT to treat mycoses or kill fungi in the environment depends on identifying effective PS for the different pathogenic species and delivery systems able to expand and optimize their use. In the present study, the in vitro susceptibility of Cryptococcus neoformans melanized cells to the photodynamic effects of the PS agent ClAlPc in nanoemulsion (ClAlPc/NE) was examined. Cells were killed in a PS concentration- and light dose-dependent manner. Treatment with ClAlPc/NE, using PS concentrations (e.g. 4.5 mu m) and light doses (e.g. 10 J cm-2) compatible with PACT, resulted in a reduction of up to 6 logs in survival. Washing the cells to remove unbound PS before light exposure did not inhibit fungal photodynamic inactivation. Internalization of ClAlPc by C.neoformans was confirmed by confocal fluorescence microscopy, and the degree of uptake was dependent on PS concentration.

COMPARISON OF PRIMERS FOR RAPD-PCR FROM ENVIRONMENTAL ISOLATES OF CRYPTOCOCCUS NEOFORMANS, CRYPTOCOCCUS ALBIDUS AND CRYPTOCOCCUS LAURENTII COMPLEX

Various organisms have been characterized by molecular methods, including fungi of the genus Cryptococcus. The purposes of this study were: to determine the discriminatory potential of the RAPD (Random Amplified Polymorphic DNA) primers, the pattern of similarity of the Cryptococcus species, and discuss their useful application in epidemiological studies. We analyzed 10 isolates of each specie/group: C. albidus, C. laurentii complex, C. neoformans var. grubii, all from environmental source, and two ATCC strains, C. neoformans var. grubii ATCC 90112, and C. neoformans var. neoformans ATCC 28957 by RAPD-PCR using the primers CAV1, CAV2, ZAP19, ZAP20, OPB11 and SEQ6. The primers showed a good discriminatory power, revealing important differences between them and between species; the SEQ6 primer discriminated a larger number of isolates of three species. Isolates of C. laurentii showed greater genetic diversity than other species revealed by all six primers. Isolates of C. neoformans were more homogeneous. Only the primer CAV2 showed no amplification of DNA bands for C. albidus. It was concluded that the use of limited number of carefully selected primers allowed the discrimination of different isolates, and some primers (e. g., CAV2 for C. albidus) may not to be applied to some species.

Comparison of primers for RAPD-PCR from environmental isolates of Cryptococcus neoformans, Cryptococcus albidus and Cryptococcus laurentii complex

Various organisms have been characterized by molecular methods, including fungi of the genus Cryptococcus. The purposes of this study were: to determine the discriminatory potential of the RAPD (Random Amplified Polymorphic DNA) primers, the pattern of similarity of the Cryptococcus species, and discuss their useful application in epidemiological studies. We analyzed 10 isolates of each specie/group: C. albidus, C. laurentii complex, C. neoformans var. grubii, all from environmental source, and two ATCC strains, C. neoformans var. grubii ATCC 90112, and C. neoformans var. neoformans ATCC 28957 by RAPD-PCR using the primers CAV1, CAV2, ZAP19, ZAP20, OPB11 and SEQ6. The primers showed a good discriminatory power, revealing important differences between them and between species; the SEQ6 primer discriminated a larger number of isolates of three species. Isolates of C. laurentii showed greater genetic diversity than other species revealed by all six primers. Isolates of C. neoformans were more homogeneous. Only the primer CAV2 showed no amplification of DNA bands for C. albidus. It was concluded that the use of limited number of carefully selected primers allowed the discrimination of different isolates, and some primers (e.g., CAV2 for C. albidus) may not to be applied to some species.