Vaccine development for pathogenic fungi: current status and future directions.

INTRODUCTION: Fungal infections are caused by a broad range of pathogenic fungi that are found worldwide with different geographic distributions, incidences, and mortality rates. Considering that there are relatively few approved medications available for combating fungal diseases and no vaccine formulation commercially available, multiple groups are searching for new antifungal drugs, examining drugs for repurposing and developing antifungal vaccines, in order to control deaths, sequels, and the spread of these complex infections. AREAS COVERED: This review provides a summary of advances in fungal vaccine studies and the different approaches under development, such as subunit vaccines, whole organism vaccines, and DNA vaccines, as well as studies that optimize the use of adjuvants. We conducted a literature search of the PubMed with terms: fungal vaccines and genus of fungal pathogens (Cryptococcus spp. Candida spp. Coccidioides spp. Aspergillus spp. Sporothrix spp. Histoplasma spp. Paracoccidioides spp. Pneumocystis spp. and the Mucorales order), a total of 177 articles were collected from database. EXPERT OPINION: Problems regarding the immune response development in an immunocompromised organism, the similarity between fungal and mammalian cells, and the lack of attention by health organizations to fungal infections are closely related to the fact that, at present, there are no fungal vaccines available for clinical use.

Deletion of AA9 Lytic Polysaccharide Monooxygenases Impacts A. nidulans Secretome and Growth on Lignocellulose.

Lytic polysaccharide monooxygenases (LPMOs) are oxidative enzymes found in viruses, archaea, and bacteria as well as eukaryotes, such as fungi, algae and insects, actively contributing to the degradation of different polysaccharides. In Aspergillus nidulans, LPMOs from family AA9 (AnLPMO9s), along with an AA3 cellobiose dehydrogenase (AnCDH1), are cosecreted upon growth on crystalline cellulose and lignocellulosic substrates, indicating their role in the degradation of plant cell wall components. Functional analysis revealed that three target LPMO9s (AnLPMO9C, AnLPMO9F and AnLPMO9G) correspond to cellulose-active enzymes with distinct regioselectivity and activity on cellulose with different proportions of crystalline and amorphous regions. AnLPMO9s deletion and overexpression studies corroborate functional data. The abundantly secreted AnLPMO9F is a major component of the extracellular cellulolytic system, while AnLPMO9G was less abundant and constantly secreted, and acts preferentially on crystalline regions of cellulose, uniquely displaying activity on highly crystalline algae cellulose. Single or double deletion of AnLPMO9s resulted in about 25% reduction in fungal growth on sugarcane straw but not on Avicel, demonstrating the contribution of LPMO9s for the saprophytic fungal lifestyle relies on the degradation of complex lignocellulosic substrates. Although the deletion of AnCDH1 slightly reduced the cellulolytic activity, it did not affect fungal growth indicating the existence of alternative electron donors to LPMOs. Additionally, double or triple knockouts of these enzymes had no accumulative deleterious effect on the cellulolytic activity nor on fungal growth, regardless of the deleted gene. Overexpression of AnLPMO9s in a cellulose-induced secretome background confirmed the importance and applicability of AnLPMO9G to improve lignocellulose saccharification. IMPORTANCE Fungal lytic polysaccharide monooxygenases (LPMOs) are copper-dependent enzymes that boost plant biomass degradation in combination with glycoside hydrolases. Secretion of LPMO9s arsenal by Aspergillus nidulans is influenced by the substrate and time of induction. These findings along with the biochemical characterization of novel fungal LPMO9s have implications on our understanding of their concerted action, allowing rational engineering of fungal strains for biotechnological applications such as plant biomass degradation. Additionally, the role of oxidative players in fungal growth on plant biomass was evaluated by deletion and overexpression experiments using a model fungal system.

Fungal Extracellular Vesicles Are Involved in Intraspecies Intracellular Communication.

Fungal infections are associated with high mortality rates in humans. The risk of fungal diseases creates the urgent need to broaden the knowledge base regarding their pathophysiology. In this sense, the role of extracellular vesicles (EVs) has been described to convey biological information and participate in the fungus-host interaction process. We hypothesized that fungal EVs work as an additional element in the communication routes regulating fungal responses in intraspecies interaction systems. In this respect, the aim of this study was to address the gene regulation profiles prompted by fungal EVs in intraspecies recipient cells. Our data demonstrated the intraspecies uptake of EVs in pathogenic fungi, such as Candida albicans, Aspergillus fumigatus, and Paracoccidioides brasiliensis, and the effects triggered by EVs in fungal cells. In C. albicans, we evaluated the involvement of EVs in the yeast-to-hypha transition, while in P. brasiliensis and A. fumigatus the function of EVs as stress transducers was investigated. P. brasiliensis and A. fumigatus were exposed to an inhibitor of glycosylation or UV light, respectively. The results demonstrated the role of EVs in regulating the expression of target genes and triggering phenotypic changes. The EVs treatment induced cellular proliferation and boosted the yeast to hyphal transition in C. albicans, while they enhanced stress responsiveness in A. fumigatus and P. brasiliensis, establishing a role for EVs in fungal intraspecies communication. Thus, EVs regulate fungal behavior, acting as potent message effectors, and understanding their effects and mechanism(s) of action could be exploited in antifungal therapies. IMPORTANCE Here, we report a study about extracellular vesicles (EVs) as communication mediators in fungi. Our results demonstrated the role of EVs from Candida albicans, Aspergillus fumigatus, and Paracoccidioides brasiliensis regulating the expression of target genes and phenotypic features. We asked whether fungal EVs play a role as message effectors. We show that fungal EVs are involved in fungal interaction systems as potent message effectors, and understanding their effects and mechanisms of action could be exploited in antifungal therapies.

Perception of fungi by farmers in the Cerrado / Percepção de agricultores do Cerrado sobre os fungos

Fungi are important in several aspects of human life. In particular, to agriculture, pathogenic fungi are of great importance, as they are responsible for production losses of the most diverse types. Because of this, knowledge about pathogenic fungus is of extreme importance for farmers and professionals working in agricultural areas. Among farmers who use specific agroecological practices, this knowledge is even more valuable, since by not adopting conventional methods of production, they resort to non-invasive alternatives that are less or not harmful at all to the environment in consideration of production management methods. This study aimed to assess farmer perception in the Cerrado biome in the city of Goiás (GO), Brazil, in order to understand their ethnomycological perceptions to verify historical management practices, their knowledge about phytopathogenic fungi, and how these producers perceive fungi. We used the theoretical reference method "From peasant to peasant" formulated by ANPA - National Association of Small Farmers. Some aspects of farmers' ethnomycological knowledge are discussed. These ease identification the representatives of the Fungi Kingdom is associated with organisms that present easily recognizable characteristics, such as wood-ears or disease-causing fungi. In general, farmers are able to identify representatives of the Fungi Kingdom that are found in their daily lives. The perception of farmers about fungi, a group still much unknown by society, is very relevant for future actions of ethnomycology.

Percepção dos fungos por agricultores do Cerrado – Fungos são organismos importantes em vários aspectos da vida humana. Em particular, para a agricultura, fungos patogênicos são de grande importância, pois são responsáveis por perdas de produção dos mais diversos tipos. Por isso, o conhecimento sobre fungos patogênicos é de extrema importância para agricultores e profissionais que trabalham em áreas agrícolas. Entre os agricultores que utilizam práticas agroecológicas, esse conhecimento é ainda mais valioso, pois, ao não adotar métodos convencionais de produção, recorrem a alternativas não invasivas que são menos ou não prejudiciais ao meio ambiente, considerando os métodos de gerenciamento da produção. Este estudo teve como objetivo avaliar a percepção de um grupo de agricultores do bioma Cerrado, na cidade de Goiás (GO), Brasil, a fim de compreender suas percepções etnomicológicas, de modo a verificar práticas históricas de manejo, seu conhecimento sobre fungos fitopatogênicos e como esses produtores percebem os fungos, de modo geral. Utilizamos o método "de camponês para camponês", formulado pela ANPA – Associação Nacional de Pequenos Agricultores. Discutimos alguns aspectos do conhecimento etnomicológico destes agricultores. A facilidade de identificação de representantes fúngicos está associada a organismos que apresentam características facilmente reconhecíveis, como estereótipo orelha-de-pau ou fungos causadores de doenças em cultivares. Em geral, os agricultores do estudo são capazes de identificar representantes do Reino Fungi que são encontrados em suas vidas cotidianas. A percepção dos agricultores sobre os fungos, um grupo ainda muito desconhecido pela sociedade, é muito relevante para ações futuras dentro da etnomicologia.

Perception of fungi by farmers in the Cerrado

Abstract Fungi are important in several aspects of human life. In particular, to agriculture, pathogenic fungi are of great importance, as they are responsible for production losses of the most diverse types. Because of this, knowledge about pathogenic fungus is of extreme importance for farmers and professionals working in agricultural areas. Among farmers who use specific agroecological practices, this knowledge is even more valuable, since by not adopting conventional methods of production, they resort to non-invasive alternatives that are less or not harmful at all to the environment in consideration of production management methods. This study aimed to assess farmer perception in the Cerrado biome in the city of Goiás (GO), Brazil, in order to understand their ethnomycological perceptions to verify historical management practices, their knowledge about phytopathogenic fungi, and how these producers perceive fungi. We used the theoretical reference method From peasant to peasant formulated by ANPA - National Association of Small Farmers. Some aspects of farmers ethnomycological knowledge are discussed. These ease identification the representatives of the Fungi Kingdom is associated with organisms that present easily recognizable characteristics, such as wood-ears or disease-causing fungi. In general, farmers are able to identify representatives of the Fungi Kingdom that are found in their daily lives. The perception of farmers about fungi, a group still much unknown by society, is very relevant for future actions of ethnomycology.

Resumo Percepção dos fungos por agricultores do Cerrado Fungos são organismos importantes em vários aspectos da vida humana. Em particular, para a agricultura, fungos patogênicos são de grande importância, pois são responsáveis por perdas de produção dos mais diversos tipos. Por isso, o conhecimento sobre fungos patogênicos é de extrema importância para agricultores e profissionais que trabalham em áreas agrícolas. Entre os agricultores que utilizam práticas agroecológicas, esse conhecimento é ainda mais valioso, pois, ao não adotar métodos convencionais de produção, recorrem a alternativas não invasivas que são menos ou não prejudiciais ao meio ambiente, considerando os métodos de gerenciamento da produção. Este estudo teve como objetivo avaliar a percepção de um grupo de agricultores do bioma Cerrado, na cidade de Goiás (GO), Brasil, a fim de compreender suas percepções etnomicológicas, de modo a verificar práticas históricas de manejo, seu conhecimento sobre fungos fitopatogênicos e como esses produtores percebem os fungos, de modo geral. Utilizamos o método de camponês para camponês, formulado pela ANPA Associação Nacional de Pequenos Agricultores. Discutimos alguns aspectos do conhecimento etnomicológico destes agricultores. A facilidade de identificação de representantes fúngicos está associada a organismos que apresentam características facilmente reconhecíveis, como estereótipo orelha-de-pau ou fungos causadores de doenças em cultivares. Em geral, os agricultores do estudo são capazes de identificar representantes do Reino Fungi que são encontrados em suas vidas cotidianas. A percepção dos agricultores sobre os fungos, um grupo ainda muito desconhecido pela sociedade, é muito relevante para ações futuras dentro da etnomicologia.

Perception of fungi by farmers in the Cerrado / Percepção de agricultores do Cerrado sobre os fungos

Fungi are important in several aspects of human life. In particular, to agriculture, pathogenic fungi are of great importance, as they are responsible for production losses of the most diverse types. Because of this, knowledge about pathogenic fungus is of extreme importance for farmers and professionals working in agricultural areas. Among farmers who use specific agroecological practices, this knowledge is even more valuable, since by not adopting conventional methods of production, they resort to non-invasive alternatives that are less or not harmful at all to the environment in consideration of production management methods. This study aimed to assess farmer perception in the Cerrado biome in the city of Goiás (GO), Brazil, in order to understand their ethnomycological perceptions to verify historical management practices, their knowledge about phytopathogenic fungi, and how these producers perceive fungi. We used the theoretical reference method "From peasant to peasant" formulated by ANPA - National Association of Small Farmers. Some aspects of farmers' ethnomycological knowledge are discussed. These ease identification the representatives of the Fungi Kingdom is associated with organisms that present easily recognizable characteristics, such as wood-ears or disease-causing fungi. In general, farmers are able to identify representatives of the Fungi Kingdom that are found in their daily lives. The perception of farmers about fungi, a group still much unknown by society, is very relevant for future actions of ethnomycology.

Percepção dos fungos por agricultores do Cerrado ­ Fungos são organismos importantes em vários aspectos da vida humana. Em particular, para a agricultura, fungos patogênicos são de grande importância, pois são responsáveis por perdas de produção dos mais diversos tipos. Por isso, o conhecimento sobre fungos patogênicos é de extrema importância para agricultores e profissionais que trabalham em áreas agrícolas. Entre os agricultores que utilizam práticas agroecológicas, esse conhecimento é ainda mais valioso, pois, ao não adotar métodos convencionais de produção, recorrem a alternativas não invasivas que são menos ou não prejudiciais ao meio ambiente, considerando os métodos de gerenciamento da produção. Este estudo teve como objetivo avaliar a percepção de um grupo de agricultores do bioma Cerrado, na cidade de Goiás (GO), Brasil, a fim de compreender suas percepções etnomicológicas, de modo a verificar práticas históricas de manejo, seu conhecimento sobre fungos fitopatogênicos e como esses produtores percebem os fungos, de modo geral. Utilizamos o método "de camponês para camponês", formulado pela ANPA ­ Associação Nacional de Pequenos Agricultores. Discutimos alguns aspectos do conhecimento etnomicológico destes agricultores. A facilidade de identificação de representantes fúngicos está associada a organismos que apresentam características facilmente reconhecíveis, como estereótipo orelha-de-pau ou fungos causadores de doenças em cultivares. Em geral, os agricultores do estudo são capazes de identificar representantes do Reino Fungi que são encontrados em suas vidas cotidianas. A percepção dos agricultores sobre os fungos, um grupo ainda muito desconhecido pela sociedade, é muito relevante para ações futuras dentro da etnomicologia.

Perception of fungi by farmers in the Cerrado / Percepção de agricultores do Cerrado sobre os fungos

Fungi are important in several aspects of human life. In particular, to agriculture, pathogenic fungi are of great importance, as they are responsible for production losses of the most diverse types. Because of this, knowledge about pathogenic fungus is of extreme importance for farmers and professionals working in agricultural areas. Among farmers who use specific agroecological practices, this knowledge is even more valuable, since by not adopting conventional methods of production, they resort to non-invasive alternatives that are less or not harmful at all to the environment in consideration of production management methods. This study aimed to assess farmer perception in the Cerrado biome in the city of Goiás (GO), Brazil, in order to understand their ethnomycological perceptions to verify historical management practices, their knowledge about phytopathogenic fungi, and how these producers perceive fungi. We used the theoretical reference method "From peasant to peasant" formulated by ANPA - National Association of Small Farmers. Some aspects of farmers' ethnomycological knowledge are discussed. These ease identification the representatives of the Fungi Kingdom is associated with organisms that present easily recognizable characteristics, such as wood-ears or disease-causing fungi. In general, farmers are able to identify representatives of the Fungi Kingdom that are found in their daily lives. The perception of farmers about fungi, a group still much unknown by society, is very relevant for future actions of ethnomycology.(AU)

Percepção dos fungos por agricultores do Cerrado Fungos são organismos importantes em vários aspectos da vida humana. Em particular, para a agricultura, fungos patogênicos são de grande importância, pois são responsáveis por perdas de produção dos mais diversos tipos. Por isso, o conhecimento sobre fungos patogênicos é de extrema importância para agricultores e profissionais que trabalham em áreas agrícolas. Entre os agricultores que utilizam práticas agroecológicas, esse conhecimento é ainda mais valioso, pois, ao não adotar métodos convencionais de produção, recorrem a alternativas não invasivas que são menos ou não prejudiciais ao meio ambiente, considerando os métodos de gerenciamento da produção. Este estudo teve como objetivo avaliar a percepção de um grupo de agricultores do bioma Cerrado, na cidade de Goiás (GO), Brasil, a fim de compreender suas percepções etnomicológicas, de modo a verificar práticas históricas de manejo, seu conhecimento sobre fungos fitopatogênicos e como esses produtores percebem os fungos, de modo geral. Utilizamos o método "de camponês para camponês", formulado pela ANPA Associação Nacional de Pequenos Agricultores. Discutimos alguns aspectos do conhecimento etnomicológico destes agricultores. A facilidade de identificação de representantes fúngicos está associada a organismos que apresentam características facilmente reconhecíveis, como estereótipo orelha-de-pau ou fungos causadores de doenças em cultivares. Em geral, os agricultores do estudo são capazes de identificar representantes do Reino Fungi que são encontrados em suas vidas cotidianas. A percepção dos agricultores sobre os fungos, um grupo ainda muito desconhecido pela sociedade, é muito relevante para ações futuras dentro da etnomicologia.(AU)

Comparative Proteomic Analysis of Histoplasma capsulatum Yeast and Mycelium Reveals Differential Metabolic Shifts and Cell Wall Remodeling Processes in the Different Morphotypes.

Histoplasma capsulatum is a thermally dimorphic fungus distributed worldwide, but with the highest incidence in the Americas within specific geographic areas, such as the Mississippi River Valley and regions in Latin America. This fungus is the etiologic agent of histoplasmosis, an important life-threatening systemic mycosis. Dimorphism is an important feature for fungal survival in different environments and is related to the virulence of H. capsulatum, and essential to the establishment of infection. Proteomic profiles have made important contributions to the knowledge of metabolism and pathogenicity in several biological models. However, H. capsulatum proteome studies have been underexplored. In the present study, we report the first proteomic comparison between the mycelium and the yeast cells of H. capsulatum. Liquid chromatography coupled to mass spectrometry was used to evaluate the proteomic profile of the two phases of H. capsulatum growth, mycelium, and yeast. In summary, 214 and 225 proteins were only detected/or preferentially abundant in mycelium or yeast cells, respectively. In mycelium, enzymes related to the glycolytic pathway and to the alcoholic fermentation occurred in greater abundance, suggesting a higher use of anaerobic pathways for energy production. In yeast cells, proteins related to the tricarboxylic acid cycle and response to temperature stress were in high abundance. Proteins related to oxidative stress response or involved with cell wall metabolism were identified with differential abundance in both conditions. Proteomic data validation was performed by enzymatic activity determination, Western blot assays, or immunofluorescence microscopy. These experiments corroborated, directly or indirectly, the abundance of isocitrate lyase, 2-methylcitrate synthase, catalase B, and mannosyl-oligosaccharide-1,2-alpha-mannosidase in the mycelium and heat shock protein (HSP) 30, HSP60, glucosamine-fructose-6-phosphate aminotransferase, glucosamine-6-phosphate deaminase, and N-acetylglucosamine-phosphate mutase in yeast cells. The proteomic profile-associated functional classification analyses of proteins provided new and interesting information regarding the differences in metabolism between the two distinct growth forms of H. capsulatum.

Fungal endophytes in germinated seeds of the common bean, Phaseolus vulgaris.

We conducted a survey of fungal endophytes in 582 germinated seeds belonging to 11 Colombian cultivars of the common bean (Phaseolus vulgaris). The survey yielded 394 endophytic isolates belonging to 42 taxa, as identified by sequence analysis of the ribosomal DNA internal transcribed spacer (ITS) region. Aureobasidium pullulans was the dominant endophyte, isolated from 46.7 % of the samples. Also common were Fusarium oxysporum, Xylaria sp., and Cladosporium cladosporioides, but found in only 13.4 %, 11.7 %, and 7.6 % of seedlings, respectively. Endophytic colonization differed significantly among common bean cultivars and seedling parts, with the highest colonization occurring in the first true leaves of the seedlings.