Scedosporium apiospermum, Scedosporium aurantiacum, Scedosporium minutisporum and Lomentospora prolificans: a comparative study of surface molecules produced by conidial and germinated conidial cells

BACKGROUND Scedosporium/Lomentospora species are opportunistic mould pathogens, presenting notable antifungal resistance. OBJECTIVES/METHODS We analysed the conidia and germinated conidia of S. apiospermum (Sap), S. aurantiacum (Sau), S. minutisporum (Smi) and L. prolificans (Lpr) by scanning electron microscopy and exposition of surface molecules by fluorescence microscopy. FINDINGS Conidia of Sap, Smi and Sau had oval, ellipsoidal and cylindrical shape, respectively, with several irregularities surrounding all surface areas, whereas Lpr conidia were rounded with a smooth surface. The germination of Sap occurred at the conidial bottom, while Smi and Sau germination primarily occurred at the centre of the conidial cell, and Lpr germination initiated at any part of the conidial surface. The staining of N-acetylglucosamine-containing molecules by fluorescein-labelled WGA primarily occurred during the germination of all studied fungi and in the conidial scars, which is the primary location of germination. Calcofluor white, which recognises the polysaccharide chitin, strongly stained the conidial cells and, to a lesser extent, the germination. Both mannose-rich glycoconjugates (evidenced by fluoresceinated-ConA) and cell wall externally located polypeptides presented distinct surface locations and expression according to both morphotypes and fungal species. In contrast, sialic acid and galactose-containing structures were not detected at fungal surfaces. MAIN CONCLUSIONS The present study demonstrated the differential production/exposition of surface molecules on distinct morphotypes of Scedosporium/Lomentospora species.

Conidial germination in Scedosporium apiospermum, S. aurantiacum, S. minutisporum and Lomentospora prolificans: influence of growth conditions and antifungal susceptibility profiles

In the present study, we have investigated some growth conditions capable of inducing the conidial germination in Scedosporium apiospermum, S. aurantiacum, S. minutisporum and Lomentospora prolificans. Germination in Sabouraud medium (pH 7.0, 37ºC, 5% CO2) showed to be a typically time-dependent event, reaching ~75% in S. minutisporum and > 90% in S. apiospermum, S. aurantiacum and L. prolificans after 4 h. Similar germination rate was observed when conidia were incubated under different media and pHs. Contrarily, temperature and CO2 tension modulated the germination. The isotropic conidial growth (swelling) and germ tube-like projection were evidenced by microscopy and cytometry. Morphometric parameters augmented in a time-dependent fashion, evidencing changes in size and granularity of fungal cells compared with dormant 0 h conidia. In parallel, a clear increase in the mitochondrial activity was measured during the transformation of conidia-into-germinated conidia. Susceptibility profiles to itraconazole, fluconazole, voriconazole, amphotericin B and caspofungin varied regarding each morphotype and each fungal species. Overall, the minimal inhibitory concentrations for hyphae were higher than conidia and germinated conidia, except for caspofungin. Collectively, our study add new data about the conidia-into-hyphae transformation in Scedosporium and Lomentospora species, which is a relevant biological process of these molds directly connected to their antifungal resistance and pathogenicity mechanisms.

THE POWER OF THE SMALL: THE EXAMPLE OF Paracoccidioides brasiliensis CONIDIA / O poder do pequeno: o exemplo dos conídios do Paracoccidioides brasiliensis

SUMMARYResearch on Paracoccidioides brasiliensis has centered in the yeast cell probably because of the lack of distinctive features in the mycelium. In 1942 and for the first time, lateral conidia were noticed in the fungus' hyphae. Later on, Brazilian, Venezuelan and Argentinean researchers described "aleurias" when the fungus was grown in natural substrates. In 1970 authors became interested in the conidia and were able to obtain them in large numbers and treat them as individual units. Their shape and size were defined and the presence of all the elements of a competent eukaryotic cell were demonstrated. Conidia exhibited thermal dimorphism and, additionally, when given intranasally to BALB/c male mice, they converted into yeasts in the lungs and produce progressive pulmonary lesions with further dissemination to other organs. Studies on the phagocyte-conidia interaction were revealing and showed that these versatile structures allow a better understanding of the host- P. brasiliensisinteractions.

RESUMOA investigação sobre Paracoccidioides brasiliensis tem-se centrado na célula de levedura, provavelmente devido à falta de características distintas no micélio. Em 1942 e, pela primeira vez, conídios laterais foram notados nos hifas dos fungos. Mais tarde, pesquisadores brasileiros, venezuelanos e argentinos descreveram "aleurias" quando o fungo foi cultivado em substratos naturais. Em 1970, os autores se interessaram pelos conídios e foram capazes de obtê-los em grande número e tratá-los como unidades individuais. A sua forma e tamanho foram definidos, e a presença de todos os elementos de uma célula eucariótica competente foram demonstrados. Conídios apresentam dimorfismo térmico e, além disso, quando administrados por via intranasal a camundongos BALB/c machos, são convertidos em leveduras nos pulmões e produzem lesões pulmonares progressivas com posterior disseminação para outros órgãos. Estudos sobre a interação de fagócitos-conídios foram reveladores e mostraram que estas estruturas versáteis permitem melhor compreensão das interacções entre hospedeiro e P. brasiliensis.

Interaction between Paracoccidioides brasiliensis conidia and the coagulation system: involvement of fibrinogen

The infectious process starts with an initial contact between pathogen and host. We have previously demonstrated that Paracoccidioides brasiliensis conidia interact with plasma proteins including fibrinogen, which is considered the major component of the coagulation system. In this study, we evaluated the in vitro capacity of P. brasiliensis conidia to aggregate with plasma proteins and compounds involved in the coagulation system. We assessed the aggregation of P. brasiliensis conidia after incubation with human serum or plasma in the presence or absence of anticoagulants, extracellular matrix (ECM) proteins, metabolic and protein inhibitors, monosaccharides and other compounds. Additionally, prothrombin and partial thromboplastin times were determined after the interaction of P. brasiliensis conidia with human plasma. ECM proteins, monosaccharides and human plasma significantly induced P. brasiliensis conidial aggregation; however, anticoagulants and metabolic and protein inhibitors diminished the aggregation process. The extrinsic coagulation pathway was not affected by the interaction between P. brasiliensis conidia and plasma proteins, while the intrinsic pathway was markedly altered. These results indicate that P. brasiliensis conidia interact with proteins involved in the coagulation system. This interaction may play an important role in the initial inflammatory response, as well as fungal disease progression caused by P. brasiliensis dissemination.

Efecto de la calidad del agua de criaderos de mosquitos (Diptera: Culicidae) sobre la patogenicidad e infectividad de las zoosporas del hongo Leptolegnia chapmanii (Straminipila: Peronosporomycetes)

Effect of water quality in mosquito breeding sites on the pathogenicity and infectivity of zoospores from the fungus Leptolegnia chapmanii (Straminipila: Peronosporomycetes). The fungus Leptolegnia chapmanii is highly pathogenic to mosquito larvae in Argentina. We studied if physical and chemical characteristics of the water from mosquito breeding sites affect pathogenicity, and the infectivity of zoospores of L. chapmanii. Water samples were taken from pools filled by rains, urban ditches with domestic waste water, pools filled by overflow from Río de la Plata, and flower vases from the Cemetery of La Plata city. Sub-samples of water were analyzed for physical and chemical characteristics, while other sub-samples were used for laboratory bioassays. Containers with 150 ml of water samples, 25 Aedes aegypti larvae, and 2.8 x 105 zoospores of L. chapmanii, were incubated under controlled environment, and larval mortality was recorded after 48 h. There were highly significant differences among mortalities in water from cemetery vases (70.2%), rain pools water (99.5%), and pools with water from Rio de la Plata (95%). There were no significant differences among larval mortalities in water from ditches, rain pools and Río de la Plata pools. Leptolegnia chapmanii was successful as a biological control agent in all kinds of tested water qualities, producing high larval mortality. Rev. Biol. Trop. 57 (1-2): 371-380. Epub 2009 June 30.

Leptolegnia chapmanii es un potente patógeno de larvas de mosquitos. El objetivo del trabajo fue determinar si las características físico-químicas del agua de criaderos de culícidos de importancia sanitaria a nivel mundial afectan la patogenicidad e infectividad de las zoosporas de L. chapmanii. Se tomaron muestras de cuatro tipos de agua en Argentina: de lluvia, de zanja, del Río de La Plata, y de floreros del cementerio local. A una parte del agua se le realizaron estudios físicos-químicos, y otra parte fue utilizada en ensayos. Se colocó 150 ml de cada muestra en recipientes plásticos, con 25 larvas sanas de Aedes aegypti y 2.8 x 105 zoosporas de L. chapmanii. Fueron incubadas a 25 ºC y 12-12 (L-O). La mortalidad larval fue registrada a las 48 horas. Se utilizaron tres recipientes y un control por tratamiento, con tres repeticiones realizadas en distintos días. Se observaron diferencias altamente significativas entre la mortalidad larval en aguas de floreros del cementerio (70.2%), aguas de lluvias (99.53%), y del río de La Plata (95%). No hubo diferencias significativas entre la mortalidad larval en agua de zanja, lluvia y río. Leptolegnia chapmanii resultó exitoso en los cuatro tipos de aguas evaluados, produciendo elevados niveles de mortalidad larval.

Efecto de la heterogeneidad espacial y estacional del suelo sobre la abundancia de esporas de hongos micorrizógenos arbusculares en el valle semiárido de Tehuacán-Cuicatlán, México / Effect of the spatial and seasonal soil heterogeneity over arbuscular mycorrhizal fungal spore abundance in the semi-arid valley of Tehuacán-Cuicatlán, Mexico

Estudios recientes señalan que algunas especies de Mimosa (Leguminosae-Mimosoideae) forman islas de recursos (IR), ricas en materia orgánica y nutrientes del suelo, así como con esporas de hongos micorrizógenos arbusculares (HMA), en el Valle semiárido de Tehuacán-Cuicatlán. La importancia de este hecho radica en que las zonas áridas y semiáridas se caracterizan por la baja fertilidad del suelo y por su escasa precipitación, lo que limita el crecimiento y desarrollo de las especies vegetales, de manera que la presencia de HMA puede ser una ventaja para las plantas desérticas que forman micorrizas. Las fluctuaciones en el número de esporas de HMA del suelo pueden estar relacionadas con factores ambientales, edáficos y estacionales que afectan la esporulación de estos hongos, así como la historia de vida de la planta hospedera. El objetivo de este estudio fue evaluar el impacto de la heterogeneidad espacial (IR vs áreas abiertas, AA) y estacional (estación de lluvias vs inicio de sequía vs sequía) del suelo sobre la distribución y abundancia del número de esporas de HMA, en cuatro sitios de estudio en el Valle de Tehuacán-Cuicatlán. Se registraron esporas de HMA en las 120 muestras de suelo analizadas. Se registraron diferencias significativas al comparar entre el número de esporas de HMA del suelo bajo el dosel de las especies de Mimosa (IR) y el de las AA (IR > AA), así como al comparar entre los tres puntos de muestreo de suelo de las IR (suelo cercano al tronco > suelo debajo de la parte media de la copa > suelo debajo del margen de la copa). Sin embargo, no se registró diferencia significativa entre el número de esporas del suelo cercano al tronco y el del medio, así como entre el suelo del margen y el del AA. Finalmente, se reportó un mayor número de esporas en el suelo durante la estación de lluvias que durante la estación de sequía (lluvias > inicio de sequía > sequía). Por tanto, la distribución de esporas de HMA...

Recent studies have shown that some species of Mimosa (Leguminosae-Mimosoideae) create resource islands (RI), rich in soil organic matter and nutrients, as well as in arbuscular mycorrhyzal fungal (AMF) spores, in the semi-arid Valley of Tehuacán-Cuicatlán. The relevance of this fact is that arid and semi-arid regions are characterized by low fertility soils and scarce precipitation, limiting plant species growth and development; this explains why the presence of AM fungi may be advantageous for mycorrhizal desert plants. Fluctuations in AMF spore numbers could be related to environmental, seasonal and soil factors which affect AMF sporulation, in addition to the life history of the host plant. The aim of this study was to asses the impact of spatial (resource islands vs open areas, OA) and seasonal (wet season vs start of dry season vs dry season) soil heterogeneity in the distribution and abundance of AMF spores in four different study sites within the Valley. We registered AMF spores in the 120 soil samples examined. Significant differences in the number of AMF spores were reported in the soil below the canopy of Mimosa species (RI) comparing with OA (RI > OA), and between Mimosa RI themselves when comparing along a soil gradient within the RI (soil near the trunk > soil below the middle of the canopy > soil in the margin of the canopy > OA); however, there were no significant differences between the soil closest to the trunk vs middle, and margin vs OA. Finally, more spores were reported in the soil collected during the wet season than during the dry season (wet > start of dry > dry). Therefore, the distribution of AMF spores is affected by spatial and seasonal soil heterogeneity. This study points out the relevance of Mimosa RI as AMF spore reservoirs and the potential importance of AM fungi for plant species survivorship and establishment in semi-arid regions. AM fungi have recently...

Auxotrophic requirement for sporangiospore--yeast transformation of Dimorphomyces diastaticus strain C 12.

The filamentous microorganism tentatively known as Dimorphomyces diastaticus originally isolated from fermenting soursop extract, in contradistinction grew in the considiogeneous form in glucose-ammonium sulphate-basal salts buffered medium which induced polar budding yeast cells of another dimorphic microorganism, D. pleomorphis (1). Further experiments using this medium showed that a number of growth factors including inositol, thymine and uracil separately incorporated inducted yeast morphology. Carbon substrates gave rise to polar budding yeast cells in the following order: galactose, malic acid, maltose, but mycelial fragments were preponderant in the reverse order. There was no growth with citric acid as substrate. With maltose-substrate and uracil incorporation in buffered basal salts medium, only polar budding yeast cells were induced, while granular particles abound with thymine supplementation. It was concluded that D. diastaticus strain C12 auxotrophically required a dissacharide, maltose, and pyrimidine base, uracil, for sporangiospore-yeast transformation in buffered ammonium sulphate-basal salts medium, pH 3.5.