ABSTRACT
To re-identify and further group 25 isolates of Trichosporon spp. identified morphologically previously, sequences of D1/D2 region of large subunit (LSU) of ribosomal DNA (rDNA) of 25 tested strains for identification and those of ribosomal intergenic space 1 (IGS1) region of 11 strains for sub-grouping were detected. The identifications of tested strains were changed except 6 strains. According to the alignment of the IGS1 region, 6 T. asahii isolates tested fell into 4 groups and 5 T. faecale isolates into 3 groups. Polymorphism of 2 T.japonicum isolates was found in 10 positions. With the alignments obtained in this research compared with the relative GenBank entries, it was found that T. asahii, T.faecale and T.japonicum species were divided into 7, 3 and 2 subtypes respectively. Morphological and biophysical methods are not sufficient for Trichosporon spp. identification. Sequencing becomes neces-sary for Trichosporon diagnosis. There is obvious diversity within a species.
ABSTRACT
A paracoccidioidomicose (PCM) é uma micose sistêmica, restrita à América Latina, com maior incidência noBrasil. O camundongo ddY tem sido empregado como modelo murino de PCM e, no entanto, não há informaçõesa respeito da resposta imune desse animal frente à infecção. O presente estudo tem como objetivo avaliar aresposta imune humoral específica para o principal antígeno, gp43, do fungo Paracoccidioides brasiliensis,em camundongos ddY infectados com a cepa virulenta Pb 18. Foram realizadas análises da antigenemia ehistopatológico em vários órgãos e em diferentes tempos pós-infecção. Os resultados obtidos demonstraramaumento nos níveis de IgG anti-gp43 nos dias 14, 17, 21, 24, 28 e 56 pós-infecção e aumento no nível de gp43solúvel aos 28 dias pós-infecção. As células fúngicas foram detectadas em todos os órgãos analisados(cérebro, coração, pulmão, fígado, baço e rim) e em todos os períodos. As lesões granulomatosas tornaramsepredominantes 14 dias pós-infecção. Os resultados evidenciaram que o camundongo ddY produz respostaimune humoral frente ao principal antígeno de P. brasiliensis, apresentando-se elevado até 56 dias pósinfecção.A redução do nível de gp43 solúvel na fase crônica, supostamente devido ao início do controle dainfecção, requer estudos complementares adicionais.
Paracoccidioidomycosis (PCM) is a systemic mycosis, restrict to Latin America, with higher incidence inBrazil. ddY mice have been used as experimental PCM model, although there is no data regarding immuneresponse. The aim of the present study was evaluated specific humoral response against the main specificantigen of the fungal Paracoccidioides brasiliensis, the gp43, in ddY mice infected with virulent Pb 18.Antigenemia analysis and histophatological exam in several organs were performed in different time post-infection The results showed increased levels of anti-gp43 IgG on days 14, 17, 21, 24, 28 and 56 post-infectionand increased levels of soluble gp-43 on day 28 post-infection. The fungal cells were detected in all organsanalyzed (brain, heart, lung, liver, spleen and kidney) in all investigated periods. The granulomatous lesionsbecame predominant 14 days after infection. The results evidence that ddY mice produce humoral immuneresponse to main P. brasiliensis antigen, with high levels until 56 days after infection. Further studies areneeded to show that reduction of soluble gp43 in chronic phase correlates with infection control.
Subject(s)
Mice , Immunity, Mucosal , ParacoccidioidomycosisABSTRACT
Declining incidence of oropharyngeal candidosis and opportunistic infections over recent years can be attributed to the use of highly active anti-retroviral therapy (HAART). Infection with C. albicans generally involves adherence and colonization of superficial tissues. During this process, budding yeasts are able to transform to hyphae and penetrate into the deep tissue. Using the biocell tracer system, C. albicans hyphal growth was dynamically observed at the cellular level. Ritonavir was effective in the inhibition of hyphal growth with growth rate of 0.8 mum/min. This study showed the in vitro effect of HIV anti-retroviral drug on the growth rate of the C. albicans hyphae.
O declínio na incidência de candidose orofaríngea e infecções oportunistas associadas a infecção pelo HIV tem sido atribuído a introdução da terapia antiretroviral combinada (HAART). Infecção por C. albicans envolve aderência e colonização da mucosa superficial. Durante este processo leveduras são capazes de transformar-se na forma de hifas e penetrar nos tecidos mais profundos. Usando o sistema "Bio-Cell Tracer", o crescimento de hifas de C. albicans foi observado dinamicamente a nível celular. Ritonavir, inibidor de protease do HIV, foi efetivo na inibição do crescimento de hifas com media de 0.8 mim/min.O presente estudo demonstrou o efeito in vitro de um agente anti-retroviral HIV sobre o crescimento de hifas de C. albicans.
Subject(s)
AIDS-Related Opportunistic Infections , Candida albicans , HIV Protease Inhibitors , Hyphae/growth & development , Hyphae/isolation & purification , In Vitro Techniques , Methods , Sampling StudiesABSTRACT
Objectives To identify the route and time of transmission by Candida species from mothers' vagina to their neonates' mouth.Methods Specimens for fungal cultures were obtained from vaginal discharge of mothers just before delivery and also from the mouth of their offspring just after birth.Eleven mother-infant pairs were investigated.Candida species was identified based on morphology,biochemical analysis,and sequencing of D1/D2 domain of the large subunit of ribosomal DNA (LSUrDNA).Electrophoretic karyotyping (EK) and random amplified polymorphic DNA analysis (RAPD) were performed to search for DNA homology.Results Candida isolates (16 strains) from 8 mother-infant pairs were identified as Candida albicans by 100% homology of their D1/D2 sequences with reference strain C.albicans Y-12983 (GenBank access No.U45776).Similarly,4 strains from two mother-infant pairs and 2 isolates from the other pair were identified as Candida glabrata and Candida krusei,respectively,by 100% homology in sequences alignment of the domains with reference strains,C.glabrata Y-65(U44808) and C.krusei Y-5396 (U76347).The same EK profiles were found for each C.albicans or C.krusei strain pair from both mother and her neonate.Although different EK bands with various molecular size were generated for each C.glabrata isolate pair,they were still considered to be homologous based on the fact that main EK bands were identical.Each isolate pair from mother and her infant presented almost the same RAPD profile,except for one pair,isolates F7n and F7m,which showed minor diverse DNA bands.Conclusion Eleven Candida isolates from neonates have identical molecular characteristics with their mother's isolates.Vertical transmission may be the main pathway of Candida spp.from mothers to their neonates.
ABSTRACT
Objective To investigate the DNA polymorphism of Pseudallescheria boydii and Scedosporium apiospermum and to analyze the relationship between random amplification of polymorphic DNA (RAPD) patterns and geographic origins.Methods The genomic DNAs of 13 Pseudallescheria boydii strains and 18 Scedosporium apiospermum strains isolated from 5 countries were amplified with RAPD technique.Results All strains tested were classified into 31 patterns by combination of the results obtained with 3 primers.The cluster tendency was identified based on species difference,namely,P.boydii or S.apiospermum strains,however,no such cluster tendency was revealed based on geographic origins of the most of strains,by dendrogram analysis.Conclusions The infraspecific variability of P.boydii and S.apiospermum is considerable.The cluster tendency of RAPD profiles is of consistency with morphological properties of P.boydii and S.apiospermum to some degree,however,is of no correlation with geographic origins of the pathogenic strains.
ABSTRACT
Recently, some Mucorales species have been reported as protease producers. The production of extracellular proteases by Mucor circinelloides using glucose as substrate was studied. Experiments were carried out with different D-glucose concentrations (40, 60 and 80 g/L). Biomass, pH and protease activity were determined. Although biomass production had reached best yields for the medium containing D-glucose in a concentration of 80 g/L, the enzymatic production was higher when the substrate concentration was reduced to 40 g/L. The yield factor for product on cell growth and the yield factor for product on carbon substrate were higher when the microorganism grew in medium containing 40 g/L glucose. The kinetics parameters suggest that this strain seems to be promising as an alternative microorganism for protease production.
Subject(s)
Clinical Enzyme Tests , Fungi , Glucose , In Vitro Techniques , Mucorales , Protease Inhibitors , Culture Media , MethodsABSTRACT
O papel do conteúdo nutritivo do meio de cultura e de oxigênio na produçäo de clamidósporos pela fase micelial do Pracoccidioides brasiliensis foi investigado. Quatro cepas do fungo (18, Bt4, 1183, Pb9) foram cultivadas, a 25-C, em meio sólido rico e pobre em nutrientes. As cepas 18 e 1183 foram também cultivadas em anerobiose em atmosfera de nitrogênio. A cepa 18 produziu grande número de clamidósporos terminais e intercalares após 7-10 dias de cultura em meio sólido pobre em nutrientes (agar 2%, com dextrose e polipeptona 0,1%). As outras três cepas produziram número significativamente menor de esporos. A cepa 18 näo produziu clamidósporos quando cultivada em dois meios ricos em nutrientes (infusäo de cérebro e coraçäo, e agar destrose de batata). A incubaçäo anaeróbica da cepa 18 em atmosfera de nitrogênio apresentou pequeno crescimento micelial com a presença de numerosos clamidósporos. A nivel ultraestrutural, os clamidósporos apresentam um ou mais núcleos e numerosas mitocôndrias, indicativos de potencial para posterior desenvolvimento. Assim, os esporos produziram gemulaçäo múltipla 1 dia após incubaçäo a 35-C. Os resultados demonstraram que, sob condiçöes ambientais adversas, a fase micelial do P. brasiliensis produz clamidósporos em curto período de tempo. É possível que o fungo encontre condiçöes semelhantes no solo, produzindo os esporos, que poderiam desempenhar papel na propagaçäo da paracoccidioidomicose