The ability of Paracoccidioides to defend itself against reactive oxygen species (ROS) produced by host effector cells is a prerequisite to survive. To counteract these radicals, Paracoccidioides expresses, among different antioxidant enzymes, superoxide dismutases (SODs). In this study, we identified six SODs isoforms encoded by the Paracoccidioides genome. We determined gene expression levels of representative isolates of the phylogenetic lineages of Paracoccidioides spp. (S1, PS2, PS3 and Pb01-like) using quantitative RT-PCR. Assays were carried out to analyze SOD gene expression of yeast cells, mycelia cells, the mycelia-to-yeast transition and the yeast-to-mycelia germination, as well as under treatment with oxidative agents and during interaction with phagocytic cells. We observed an increased expression of PbSOD1 and PbSOD3 during the transition process, exposure to oxidative agents and interaction with phagocytic cells, suggesting that these proteins could assist in combating the superoxide radicals generated during the host-pathogen interaction. Using PbSOD1 and PbSOD3 knockdown strains we showed these genes are involved in the response of the fungus against host effector cells, particularly the oxidative stress response, and in a mouse model of infection. Protein sequence analysis together with functional analysis of knockdown strains seem to suggest that PbSOD3 expression is linked with a pronounced extracellular activity while PbSOD1 seems more related to intracellular requirements of the fungus. Altogether, our data suggests that P. brasiliensis actively responds to the radicals generated endogenously during metabolism and counteracts the oxidative burst of immune cells by inducing the expression of SOD isoforms.

Melanins are implicated in the pathogenesis of several important human diseases. This study confirmed the presence of melanin particles in Candida albicans in vitro and during infection. Dark particles were isolated from the digestion of C. albicans cultures and from infected tissue, as established by electron microscopy and immunofluorescence techniques.

Several cell wall constituents, including melanins or melanin-like compounds, have been implicated in the pathogenesis of a wide variety of microbial diseases caused by diverse species of pathogenic bacteria, fungi, and helminthes. Among these microorganisms, the dimorphic fungal pathogen Paracoccidioides brasiliensis produces melanin in its conidial and yeast forms. In the present study, melanin particles from P. brasiliensis were injected into BALB/c mice in order to produce monoclonal antibodies (MAbs). We identified five immunoglobulin G1 (IgG1) -chain and four IgM melanin-binding MAbs. The five IgG1 -chain isotypes are the first melanin-binding IgG MAbs ever reported. The nine MAbs labeled P. brasiliensis conidia and yeast cells both in vitro and in pulmonary tissues. The MAbs cross-reacted with melanin-like purified particles from other fungi and also with commercial melanins, such as synthetic and Sepia officinalis melanin. Melanization during paracoccidioidomycosis (PCM) was also further supported by the detection of IgG antibodies reactive to melanin from P. brasiliensis conidia and yeast in sera and bronchoalveolar lavage fluids from P. brasiliensisinfected mice, as well as in sera from human patients with PCM. Serum specimens from patients with other mycoses were also tested for melanin-binding antibodies by enzyme-linked immunosorbent assay, and crossreactivities were detected for melanin particles from different fungal sources. These results suggest that melanin from P. brasiliensis is an immunologically active fungal structure that activates a strong IgG humoral response in humans and mice.

Se informa un brote de histoplasmosis ocurrido en los integrantes de una familia y que comprometió a cuatro personas, dos mujeres, una niña y un hombre. El caso índice consultó por sintomatología respiratoria grave, de comienzo súbito, que requirió hospitalización. En los otros casos, la infección cursó de manera asintomática pero se puso en evidencia por la reactividad en las pruebas serológicas con histoplasmina. La búsqueda de una fuente común de contagio llevó a sospechar que era la tierra de un vivero que se había utilizado como fertilizante de las plantas caseras. Las suspensiones de las tierras de las macetas sirvieron para inocular ratones BALB/c, de cuyos órganos fue posible aislar el agente etiológico, Histoplasma capsulatum var. capsulatum. Si bien la histoplasmosis es más frecuente en ciertas ocupaciones y es propia de áreas rurales, las epidemias y los brotes son ahora comunes en áreas urbanas debido a actividades como la urbanización masiva, la tala de árboles, las demoliciones y el uso de tierras enriquecidas con abonos orgánicos (gallinaza, guano). Se llama la atención sobre el peligro que representa esta última actividad.Palabras clave: histoplasmosis, epidemiología, etiología, aislamiento y purificación, exposición ambiental, exposición por inhalación, Acute histoplasmosis outbreak in a family group: identification of infection sourceAn outbreak of acute histoplasmosis occurring in 4 members of the same family, two women, a girl and a male, is reported. The index case presented acute respiratory symptoms, severe enough to require hospitalization. In the remaining persons, the infection was asymptomatic but was evidenced by reactive histoplasmin serologic tests. Search for the common source of infection led to an enriched soil obtained in a local nursery for growing in-door plants. BALB/c mice were inoculated with suspensions of soils from the potted plants. Histoplasma capsulatum var. capsulatum was isolated from various internal organs of the mice. Although histoplasmosis is observed more frequently in persons with occupations implying risk of exposure and is connected to rural areas, outbreaks and intra-family cases are now common in urban areas. This is due to massive urbanization, deforestation, demolitions and the use of soils enriched with organic compounds, mainly bird/bat excrements. This report calls the attention on the danger involved in using such enriched soils for plant nutrition.