Second world record for two Calvatia species (Agaricaceae: Basidiomycota) / Segundo registro mundial para duas espécies de Calvatia (Agaricaceae: Basidiomycota)

Abstract Calvatia is a genus of gasteroid fungi, comprising about 47 species worldwide. In this paper we report the second worldwide occurrence of two poorly known species of Calvatia, recorded in the Cerrado biome of Brazil: C. oblongispora and C. nodulata. Detailed morphological descriptions and illustrations, including scanning electron micrographs of hyphae and basidiospores are provided, as well a discussion on their taxonomy and geographic distribution.

Resumo Calvatia é um gênero de fungos gasteroides que compreende cerca de 47 espécies em todo o mundo. Neste artigo relatamos a segunda ocorrência de duas espécies pouco conhecidas de Calvatia, registradas no bioma Cerrado do Brasil: C. oblongispora e C. nodulata. Descrições morfológicas detalhadas e ilustrações são fornecidas, incluindo micrografias eletrônicas de varredura de hifas e basidiósporos, bem como uma discussão sobre sua taxonomia e distribuição geográfica.

New records of Geastrum (Geastrales, Basidiomycota) for the Amazon / Novos registros de Geastrum (Geastrales, Basidiomycota) para a Amazônia

Fungi of the genus Geastrum are known as earthstar fungi, due to the stelliform morphology of their basidiomes. In the Amazon domain, there are 25 catalogued species of Geastrum, with several new records in recent years. Here we report three new species records of Geastrum for the Brazilian Amazon. Geastrum calycicoriaceum and G. rubropusillum were recorded in the southwestern transition zone between Amazon forest and Cerrado savanna. Geastrum calycicoriaceum was recorded for the first time in Brazil.(AU)

Os fungos do gênero Geastrum são conhecidos como fungos estrela-da-terra, devido à morfologia esteliforme de seus basidiomas. No domínio amazônico, existem 25 espécies catalogadas de Geastrum, com vários novos registos nos últimos anos. Aqui relatamos três novos registos de espécies de Geastrum para a Amazônia brasileira. Geastrum calycicoriaceum e G. rubropusillum foram registrados na zona de transição sudoeste entre a floresta amazônica e o Cerrado. Geastrum calycicoriaceum foi registado pela primeira vez no Brasil.(AU)

Characterization of wheat-Thinopyrum bessarabicum genetic stock for stripe rust and Karnal bunt resistance / Caracterização do estoque genético de trigo-Thinopyrum bessarabicum para resistência de ferrugem e Karnal bunt

Abstract Utilization of modern breeding techniques for developing high yielding and uniform plant types ultimately narrowing the genetic makeup of most crops. Narrowed genetic makeup of these crops has made them vulnerable towards disease and insect epidemics. For sustainable crop production, genetic variability of these crops must be broadened against various biotic and abiotic stresses. One of the ways to widen genetic configuration of these crops is to identify novel additional sources of durable resistance. In this regard crops wild relatives are providing valuable sources of allelic diversity towards various biotic, abiotic stress tolerance and quality components. For incorporating novel variability from wild relative's wide hybridization technique has become a promising breeding method. For this purpose, wheat-Th. bessarabicum amphiploid, addition and translocation lines have been screened in field and screen house conditions to get novel sources of yellow rust and Karnal bunt resistant. Stripe rust screening under field conditions has revealed addition lines 4JJ and 6JJ as resistant to moderately resistant while addition lines 3JJ, 5JJ, 7JJ and translocation lines Tr-3, Tr-6 as moderately resistant wheat-Thinopyrum-bessarabicum genetic stock. Karnal bunt screening depicted addition lines 5JJ and 4JJ as highly resistant genetic stock. These genetic stocks may be used to introgression novel stripe rust and Karnal bunt resistance from the tertiary gene pool into susceptible wheat backgrounds.

Resumo A utilização de técnicas modernas de melhoramento para o desenvolvimento de tipos de plantas uniformes e de alto rendimento, em última análise, estreitando a composição genética da maioria das culturas. A composição genética restrita dessas plantações tornou-as vulneráveis a doenças e epidemias de insetos. Para uma produção agrícola sustentável, a variabilidade genética dessas culturas deve ser ampliada contra vários estresses bióticos e abióticos. Uma das maneiras de ampliar a configuração genética dessas culturas é identificar novas fontes adicionais de resistência durável. A esse respeito, os parentes selvagens das culturas estão fornecendo fontes valiosas de diversidade alélica para vários componentes de qualidade e tolerância ao estresse abiótico e biótico. Para incorporar a nova variabilidade da ampla técnica de hibridização de parente selvagem tornou-se um método de reprodução promissor. Para esse efeito, trigo-Th. As linhas anfiploides, de adição e translocação de bessarabicum foram selecionadas em condições de campo e de casa de tela para obter novas fontes de ferrugem amarela e resistência ao bunt de Karnal. A triagem de ferrugem em faixas em condições de campo revelou as linhas de adição 4JJ e 6JJ como resistentes a moderadamente resistentes, enquanto as linhas de adição 3JJ, 5JJ, 7JJ e as linhas de translocação Tr-3, Tr-6 como estoque genético de trigo-Thinopyrum bessarabicum moderadamente resistente. A triagem Karnal bunt descreveu as linhas de adição 5JJ e 4JJ como estoque genético altamente resistente. Esses estoques genéticos podem ser usados para introgressão da nova ferrugem e resistência ao bunt de Karnal do pool genético terciário em origens de trigo suscetíveis.

Checklist of Amazonian gasteroid fungi (Agaricomycetidae, Phallomycetidae, Basidiomycota) / Checklist de fungos gasteroides da Amazônia (Agaricomycetidae, Phallomycetidae, Basidiomycota)

The Amazon rainforest is the largest tropical rainforest in the world, with high biodiversity of organisms. Despite the increasing knowledge about fungal diversity in the last few years, many fungal taxa, including gasteroid clades, are still poorly known. Gasteroid fungi are recognized by the maturation of basidiospores inside the basidiomata and by the passive release of spores. We conducted a detailed survey of published data on the species that occur in Amazon forests. We confirmed the occurrence of 83 species distributed in 22 genera, eight families and four orders. The most representative genera was Geastrum, followed by Cyathus, Lycoperdon and Phallus. We present an identification key for Amazonian gasteroid genera and a map of record distribution. This review is expected to contribute to identify distribution gaps for further research on gasteroid fungi and to subsidize policies for the conservation of fungi in the Amazon region.(AU)

A floresta amazônica é a maior floresta tropical do mundo, com alta biodiversidade de organismos. Apesar do crescente aumento sobre o conhecimento da diversidade fúngica nos últimos anos, muitos táxons de fungos, especialmente clados gasteróides, ainda são pouco conhecidos. Os fungos gasteróides são reconhecidos pela maturação dos basidiósporos no interior do basidioma e pela liberação passiva de esporos. Baseado em dados publicados, realizamos um levantamento detalhado das espécies que ocorrem na floresta amazônica. Nós confirmamos a ocorrência de 83 espécies distribuídas em 22 gêneros, 8 famílias e 4 ordens. Os gêneros mais representativos foram Geastrum, seguido por Cyathus, Lycoperdon e Phallus. Apresentamos uma chave de identificação para gêneros de gasteróides amazônicos e um mapa de distribuição de registros. Espera-se que esta revisão contribua para identificar lacunas de distribuição para futuras pesquisas sobre fungos gasteroides e para subsidiar políticas de conservação de fungos na região amazônica.(AU)

Two new species of rust fungi (Pucciniales) from the Brazilian Amazon: Aecidium margaritariae on Margaritaria and Uromyces amapaensis on Jatropha / Duas novas espécies de ferrugens (Pucciniales) da Amazônia brasileira: Aecidium margaritariae sobre Margaritaria e Uromyces amapaensis sobre Jatropha

Two new species of Pucciniales fungi on plants of the Fabid clade are described from samples deposited in the herbarium of Museu Paraense Emilio Goeldi, collected in the state of Amapá, in the Brazilian Amazon. They are Aecidium margaritariae found on Margaritaria nobilis (Phyllanthaceae), and Uromyces amapaensis on Jatropha gossypiifolia (Euphorbiaceae). The microstructures of the specimens were analyzed using optical and scanning electron microscopy. Aecidium margaritariae is characterized by the presence of globose, subglobose to slightly ellipsoid aeciospores with warty walls and smooth areas usually in the basal portion. Uromyces amapaensis is distinguished by the presence of uredinia with paraphyses which are thickened and rounded at the tip, and pedicellate and smooth teliospores. Descriptions, illustrations, and taxonomic comments are presented for each species.

Duas novas espécies de fungos Pucciniales sobre plantas do clado das fabídeas são descritas a partir de amostras depositadas no herbário do Museu Paraense Emílio Goeldi, coletadas no estado do Amapá, na Amazonia Brasileira. Aecidium margaritariae ocorrendo sobre Margaritaria nobilis (Phyllanthaceae) e Uromyces amapaensis sobre Jatropha gossypiifolia (Euphorbiaceae). As microestruturas dos espécimes foram analisadas em microscópio óptico e em microscopia eletrônica de varredura. Aecidium margaritariae se caracteriza por apresentar eciósporos globosos, subglobosos a levemente elipsoides, parede verrugosa com áreas lisas geralmente na extremidade basal. Uromyces amapaensis se diferencia por apresentar uredínios com paráfises engrossadas e arredondadas no ápice e teliósporos pedicelados, lisos. São apresentadas descrições, ilustrações e comentários taxonômicos para cada espécie.

Two new species of rust fungi (Pucciniales) from the Brazilian Amazon: Aecidium margaritariae on Margaritaria and Uromyces amapaensis on Jatropha / Duas novas espécies de ferrugens (Pucciniales) da Amazônia brasileira: Aecidium margaritariae sobre Margaritaria e Uromyces amapaensis sobre Jatropha

Two new species of Pucciniales fungi on plants of the Fabid clade are described from samples deposited in the herbarium of Museu Paraense Emilio Goeldi, collected in the state of Amapá, in the Brazilian Amazon. They are Aecidium margaritariae found on Margaritaria nobilis (Phyllanthaceae), and Uromyces amapaensis on Jatropha gossypiifolia (Euphorbiaceae). The microstructures of the specimens were analyzed using optical and scanning electron microscopy. Aecidium margaritariae is characterized by the presence of globose, subglobose to slightly ellipsoid aeciospores with warty walls and smooth areas usually in the basal portion. Uromyces amapaensis is distinguished by the presence of uredinia with paraphyses which are thickened and rounded at the tip, and pedicellate and smooth teliospores. Descriptions, illustrations, and taxonomic comments are presented for each species.(AU)

Duas novas espécies de fungos Pucciniales sobre plantas do clado das fabídeas são descritas a partir de amostras depositadas no herbário do Museu Paraense Emílio Goeldi, coletadas no estado do Amapá, na Amazonia Brasileira. Aecidium margaritariae ocorrendo sobre Margaritaria nobilis (Phyllanthaceae) e Uromyces amapaensis sobre Jatropha gossypiifolia (Euphorbiaceae). As microestruturas dos espécimes foram analisadas em microscópio óptico e em microscopia eletrônica de varredura. Aecidium margaritariae se caracteriza por apresentar eciósporos globosos, subglobosos a levemente elipsoides, parede verrugosa com áreas lisas geralmente na extremidade basal. Uromyces amapaensis se diferencia por apresentar uredínios com paráfises engrossadas e arredondadas no ápice e teliósporos pedicelados, lisos. São apresentadas descrições, ilustrações e comentários taxonômicos para cada espécie.(AU)

Obtaining monokaryotic and dikaryotic mycelial cultures of two Amazonian strains of Geastrum (Geastraceae, Basidiomycota) / Obtenção de culturas miceliais monocariótica e dicariótica de duas linhagens amazônicas de Geastrum (Geastraceae, Basidiomycota)

The high diversity of the genus Geastrum and the difficulty of obtaining mycelial cultures impairs the study of the ecophysiology and the exploration of the biotechnological potential of the taxon. In this study, different culture media were tested to obtain mycelial cultures for G. lloydianum and G. subiculosum collected in the Brazilian Amazon. Data on spore germination, and isolation of monokaryotic cultures and in vitro sexual reproduction are presented, as well as a brief morphological description of the cultures obtained. For both species, Potato Dextrose Agar (PDA) was the most promising of the tested culture media. The highest growth in agar culture ever recorded for this genus is reported (4.9 mm per week for G. lloydianum and 7.5 mm for G. subiculosum). In the PDA culture medium, spores germinated after 35-40 days of incubation and the isolation of monokaryotic cultures of the two species, as well as in vitro sexual crosses, were successfully performed.(AU)

A alta diversidade do gênero Geastrum e a dificuldade de obtenção de culturas miceliais prejudicam o estudo ecofisiológico e a exploração do potencial biotecnológico do táxon. Nesse estudo, foram testados diferentes meios de cultivo, visando a obtenção de culturas miceliais para G. lloydianum e G. subiculosum coletadas na Amazônia brasileira. A germinação dos esporos, o isolamento das culturas monocarióticas e o cruzamento sexual in vitro são apresentados, além de uma breve descrição morfológica das culturas obtidas. O meio de cultura Batata Dextrose Ágar (BDA) foi o mais promissor dentre os meios de cultura testados no cultivo das duas espécies. Reportamos o maior crescimento em cultura de ágar já registrado para esse gênero (4,9 mm por semana para G. lloydianum e 7,5 mm por semana para G. subiculosum). Nesse meio de cultivo, os esporos germinaram após 35-40 dias de incubação e o isolamento de culturas monocarióticas das duas espécies, assim com os cruzamentos sexuais in vitro, foram realizados com sucesso.(AU)

In vitro biological control of Ceratobasidium ramicola by using tannin extracts from Acacia villosa, Myristica fragrans, Acacia mangium, and Calliandra calothyrsus leaves / Controle biológico in vitro de Ceratobasidium ramicola usando extratos tânicos de folhas de Acacia villosa, Myristica fragrans, Acacia mangium e Calliandra calothyrsus

Ceratobasidium ramicola is a fitopathogenic fungus that harmful and causes various levels of damage on several types of forestry and horticultural crops. The purpose of this study was to examine the effect of Acacia villosa, Myristica fragrans, Acacia mangium and Calliandra calothyrsus leaf extracts as tannin sources related to the in vitro inhibition of Ceratobasidium ramicola growth. The in vitro inhibition was performed by employing solid potato dextrose agar (PDA) medium to obtain the radial inhibition, while liquid potato dextrose broth (PDB) medium was used to obtain the biomass inhibition. Experimental design was based on in time nested-completely randomized design and statistical analysis was carried out with SAS software version 9.1. The result of radial growth inhibition of Ceratobasidium ramicola showed that tannin extracts of A. mangium and M. Fragrans were not significantly different to each other. Treatment of tannin extracts from A. villosa, M. fragrans, A. mangium and C. Calothyrsus with a concentration of 1% were significantly different with other concentrations and resulted the greatest inhibition values. Tannin extract of A. Mangium at 1% concentration produced the greatest radial inhibition by 33.2%. In most cases, the effective inhibition from tannin extract occurred at the 24h of incubation. The greatest biomass inhibition was produced on 1% tannin extract of A. mangium by 64.3%, while the lowest was produced from 1% tannin extract of M. fragrans by 27.0%.(AU)

Ceratobasidium ramicola é um fungo fitopatogênico prejudicial que causa vários danos em diversas culturas florestais e agrícolas. O objetivo deste estudo foi examinar o efeito dos extratos de folhas de Acacia villosa, Myristica fragrans, Acacia mangium e Calliandra calothyrsus como fontes de tanino relacionadas à inibição do crescimento in vitro de Ceratobasidium ramicola. A inibição in vitro foi realizada empregando o meio de ágar batata dextrose para obter a inibição radial, enquanto o meio de caldo de batata dextrose líquido foi usado para obter a inibição da biomassa. O projeto experimental foi fundamentado in time. O delineamento utilizado foi inteiramente casualizado, e a análise estatística foi realizada com o software SAS, versão 9.1. O resultado da inibição do crescimento radial de Ceratobasidium ramicola mostrou que os extratos de tanino de A. mangium e M. fragrans não diferiram significativamente entre si. Os tratamentos de extratos de tanino de A. villosa, M. fragrans, A. mangium e C. Calothyrsus, utilizando uma concentração de 1%, foram significativamente diferentes em comparação com outras concentrações e resultaram em maiores valores de inibição. O extrato de tanino de A. mangium a 1% de concentração produziu a maior inibição radial, com taxa de 33,2%. Na maioria dos casos, a inibição efetiva do extrato de tanino ocorreu em 24 horas de incubação. A maior inibição de biomassa foi produzida em 1% de extrato de tanino de A. Mangium, com taxa de 64,3%, enquanto a menor foi produzida a partir de 1% de extrato de tanino de M. fragrans, com taxa de 27%.(AU)

Iron and zinc mycelial bioaccumulation in Agaricus subrufescens strains / Bioacumulação micelial de ferro e zinco em linhagens de Agaricus subrufescens

Agaricus subrufescens, a basidiomycete native to Brazil, is worldwide cultivated due to its medicinal properties. This fungus is capable of bioaccumulating metals in the mycelial biomass when cultured in the presence of them; however, this capacity is little explored for functional food production. This study aimed to evaluate the amount of iron or zinc bioaccumulated in the vegetative mycelium of sixteen strains of A. subrufescens. Mycelia were grown in malt extract agar added with 50 mg/L of iron or with 7.5 mg/L of zinc. The metal bioaccumulation in the mycelial biomass was strain-dependent. In general, metal can inhibit or stimulate the mycelial growth ranging from -81 to +78% for iron and from -86 to +100% for zinc. The highest bioaccumulated iron and zinc concentrations in the mycelial biomass was 2,595.65 mg/kg and 1,655.83 mg/kg, respectively and occurred in the U4-4 strain. The supplementation of mycelial biomass using iron or zinc is an alternative to develop food supplements, that can be used both in the human and animal diet and in the prevention of diseases.

Agaricus subrufescens, basidiomiceto nativo do Brasil, é cultivado mundialmente devido aos seus aspectos medicinais. Este fungo tem capacidade de bioacumulação de metais na biomassa micelial, no entanto, esta propriedade é pouco explorada para a produção de alimentos funcionais. O objetivo deste trabalho foi avaliar a quantidade de ferro ou zinco bioacumulada no micélio vegetativo de diversas linhagens de Agaricus subrufescens. Foram utilizadas 16 linhagens de A. subrufescens da coleção de culturas da Universidade Paranaense. O crescimento da biomassa micelial e a bioacumulação de ferro e zinco foram avaliados em micélio crescido em ágar extrato de malte adicionado de 50 mg L-1 de ferro ou 7,5 mg L-1 de zinco. A bioacumulação dos metais na biomassa micelial foi linhagem dependente. De forma geral, os metais podem inibir ou estimular o crescimento com variação entre de -81 a +78% para o ferro e de -86 a +100% para o zinco. A maior concentração de ferro na biomassa foi para o ferro de 2595,65 mg kg-1 e para o zinco de 1655,83 mg kg-1, ambos para a linhagem U4-4. O enriquecimento da biomassa micelial com ferro e/ou zinco é uma alternativa ao desenvolvimento de novos alimentos ou suplementos alimentares fonte de ferro e zinco de origem não-animal.

Iron and zinc mycelial bioaccumulation in Agaricus subrufescens strains / Bioacumulação micelial de ferro e zinco em linhagens de Agaricus subrufescens

Agaricus subrufescens, a basidiomycete native to Brazil, is worldwide cultivated due to its medicinal properties. This fungus is capable of bioaccumulating metals in the mycelial biomass when cultured in the presence of them; however, this capacity is little explored for functional food production. This study aimed to evaluate the amount of iron or zinc bioaccumulated in the vegetative mycelium of sixteen strains of A. subrufescens. Mycelia were grown in malt extract agar added with 50 mg/L of iron or with 7.5 mg/L of zinc. The metal bioaccumulation in the mycelial biomass was strain-dependent. In general, metal can inhibit or stimulate the mycelial growth ranging from -81 to +78% for iron and from -86 to +100% for zinc. The highest bioaccumulated iron and zinc concentrations in the mycelial biomass was 2,595.65 mg/kg and 1,655.83 mg/kg, respectively and occurred in the U4-4 strain. The supplementation of mycelial biomass using iron or zinc is an alternative to develop food supplements, that can be used both in the human and animal diet and in the prevention of diseases.(AU)

Agaricus subrufescens, basidiomiceto nativo do Brasil, é cultivado mundialmente devido aos seus aspectos medicinais. Este fungo tem capacidade de bioacumulação de metais na biomassa micelial, no entanto, esta propriedade é pouco explorada para a produção de alimentos funcionais. O objetivo deste trabalho foi avaliar a quantidade de ferro ou zinco bioacumulada no micélio vegetativo de diversas linhagens de Agaricus subrufescens. Foram utilizadas 16 linhagens de A. subrufescens da coleção de culturas da Universidade Paranaense. O crescimento da biomassa micelial e a bioacumulação de ferro e zinco foram avaliados em micélio crescido em ágar extrato de malte adicionado de 50 mg L-1 de ferro ou 7,5 mg L-1 de zinco. A bioacumulação dos metais na biomassa micelial foi linhagem dependente. De forma geral, os metais podem inibir ou estimular o crescimento com variação entre de -81 a +78% para o ferro e de -86 a +100% para o zinco. A maior concentração de ferro na biomassa foi para o ferro de 2595,65 mg kg-1 e para o zinco de 1655,83 mg kg-1, ambos para a linhagem U4-4. O enriquecimento da biomassa micelial com ferro e/ou zinco é uma alternativa ao desenvolvimento de novos alimentos ou suplementos alimentares fonte de ferro e zinco de origem não-animal.(AU)

Cryopreservation at -75 °C of Agaricus subrufescens on wheat grains with sucrose

Abstract Agaricus subrufescens is a basidiomycete which is studied because of its medicinal and gastronomic importance; however, less attention has been paid to its preservation. This study aimed to evaluate the effect of sucrose addition to substrate and cryotube on the viability of Agaricus subrufescens cryopreserved at -20 °C and at -75 °C for one and two years. Zero, 10% or 20% sucrose was added to potato dextrose agar or wheat grain. The mycelia were cryopreserved in the absence of cryoprotectant or with sucrose solutions at 15%, 30% or 45%. After one or two years at -75 °C or at -20 °C, mycelia were thawed and evaluated about viability, initial time of growth, colony diameter and genomic stability. Cryopreservation at -20 °C is not effective to keep mycelial viability of this fungus. Cryopreservation at -75 °C is effective when sucrose is used in substrates and/or cryotubes. Without sucrose, cryopreservation at -75 °C is effective only when wheat grains are used. Physiological characteristic as mycelial colony diameter is negatively affected when potato dextrose agar is used and unaffected when wheat grain is used after two-year cryopreservation at -75 °C. The fungus genome does not show alteration after two-year cryopreservation at -75 °C.

Long-term cryopreservation of basidiomycetes

Abstract Basidiomycetes have several biotechnological and industrial applications such as enzyme production, bioremediation, pharmaceutical and functional food production. Due to climatic features, the preservation of several basidiomycetes is threatened, and to guarantee the preservation of this genetic resource, the development of long-term preservation techniques is necessary once there is no universal protocol for the cryopreservation of basidiomycetes. Cryopreservation is a technique in which microorganisms are submitted to ultralow temperatures. Therefore, this study aimed to collect information on the main conditions for long-term cryopreservation of basidiomycetes in the last 20 years. Scientific articles on cryopreservation of basidiomycetes published from 1997 to 2016, were researched, and only the studies on two intervals of cryopreservation were considered: from 1 to 2 years and for longer than 2 years. The analyzed conditions of basidiomycete cryopreservation were: most studied genera, cryopreservation temperature, substrate, cryoprotectant (and preservation substrate), cryopreservation period, thawing temperature and cultivation medium after thawing, physiological and genetic stability of basidiomycetes after thawing in cryopreservation. In this review, the viability of the main cryopreservation conditions of basidiomycetes studied in the last 20 years are presented and discussed.

Polycyclic aromatic hydrocarbons degradation by marine-derived basidiomycetes: optimization of the degradation process

Pyrene and benzo[a]pyrene (BaP) are high molecular weight polycyclic aromatic hydrocarbons (PAHs) recalcitrant to microbial attack. Although studies related to the microbial degradation of PAHs have been carried out in the last decades, little is known about degradation of these environmental pollutants by fungi from marine origin. Therefore, this study aimed to select one PAHs degrader among three marine-derived basidiomycete fungi and to study its pyrene detoxification/degradation. Marasmiellus sp. CBMAI 1062 showed higher levels of pyrene and BaP degradation and was subjected to studies related to pyrene degradation optimization using experimental design, acute toxicity, organic carbon removal (TOC), and metabolite evaluation. The experimental design resulted in an efficient pyrene degradation, reducing the experiment time while the PAH concentration applied in the assays was increased. The selected fungus was able to degrade almost 100% of pyrene (0.08 mg mL-1) after 48 h of incubation under saline condition, without generating toxic compounds and with a TOC reduction of 17%. Intermediate metabolites of pyrene degradation were identified, suggesting that the fungus degraded the compound via the cytochrome P450 system and epoxide hydrolases. These results highlight the relevance of marine-derived fungi in the field of PAH bioremediation, adding value to the blue biotechnology.(AU)

Long-term cryopreservation of basidiomycetes

Basidiomycetes have several biotechnological and industrial applications such as enzyme production, bioremediation, pharmaceutical and functional food production. Due to climatic features, the preservation of several basidiomycetes is threatened, and to guarantee the preservation of this genetic resource, the development of long-term preservation techniques is necessary once there is no universal protocol for the cryopreservation of basidiomycetes. Cryopreservation is a technique in which microorganisms are submitted to ultralow temperatures. Therefore, this study aimed to collect information on the main conditions for long-term cryopreservation of basidiomycetes in the last 20 years. Scientific articles on cryopreservation of basidiomycetes published from 1997 to 2016, were researched, and only the studies on two intervals of cryopreservation were considered: from 1 to 2 years and for longer than 2 years. The analyzed conditions of basidiomycete cryopreservation were: most studied genera, cryopreservation temperature, substrate, cryoprotectant (and preservation substrate), cryopreservation period, thawing temperature and cultivation medium after thawing, physiological and genetic stability of basidiomycetes after thawing in cryopreservation. In this review, the viability of the main cryopreservation conditions of basidiomycetes studied in the last 20 years are presented and discussed.(AU)

Cryopreservation at -75 °C of Agaricus subrufescens on wheat grains with sucrose

Agaricus subrufescens is a basidiomycete which is studied because of its medicinal and gastronomic importance; however, less attention has been paid to its preservation. This study aimed to evaluate the effect of sucrose addition to substrate and cryotube on the viability of Agaricus subrufescens cryopreserved at -20 °C and at -75 °C for one and two years. Zero, 10% or 20% sucrose was added to potato dextrose agar or wheat grain. The mycelia were cryopreserved in the absence of cryoprotectant or with sucrose solutions at 15%, 30% or 45%. After one or two years at -75 °C or at -20 °C, mycelia were thawed and evaluated about viability, initial time of growth, colony diameter and genomic stability. Cryopreservation at -20 °C is not effective to keep mycelial viability of this fungus. Cryopreservation at -75 °C is effective when sucrose is used in substrates and/or cryotubes. Without sucrose, cryopreservation at -75 °C is effective only when wheat grains are used. Physiological characteristic as mycelial colony diameter is negatively affected when potato dextrose agar is used and unaffected when wheat grain is used after two-year cryopreservation at -75 °C. The fungus genome does not show alteration after two-year cryopreservation at -75 °C.(AU)

Dermatite fúngica associada à Rhodotorula glutinis em cão - Canis familiaris ): relato de caso / Fungal dermatitis associated to Rhodotorula glutinis in dog (Canis familiaris): case report

O fungo Rhodotorula sp. era considerado como não patogênico, mas, com o aumento de casos de humanos imunossuprimidos nas últimas duas décadas, as espécies R. mucilaginosa, R. glutinis e R. minuta se tornaram agentes potencialmente patogênicos. Poucos relatos clínicos veterinários associados à Rhodotorula spp. foram descritos, e o objetivo deste trabalho foi descrever um caso de dermatite fúngica causada por R. glutinis em cão imunossuprimido devido à leishmaniose visceral e osteossarcoma, na cidade de Cuiabá-Mato Grosso. Um cão, macho, 11 anos, sem raça definida, foi trazido ao Hospital Veterinário apresentando lesões de pele com diagnóstico de dermatofitose e foi prescrito tratamento tópico à base de óleo de melaleuca, sem melhora após um mês de tratamento. O animal retornou ao Hospital Veterinário com hemorragia na região lesionada da cauda, e foi realizada biópsia para exames microbiológico e histopatológico. Nas análises microbiológica e histopatológica, foi isolada uma levedura e detectada a presença de estrutura semelhante a leveduras na coloração de prata, respectivamente. A levedura isolada foi identificada como R. glutinis por meio do sequenciamento do DNA. Um novo tratamento foi instituído, sem melhora do quadro clínico. O animal foi diagnosticado também com leishmaniose e osteossarcoma, provavelmente ocasionando a immunossupressão, e seu quadro evoluiu a óbito.(AU)

The fungi Rhodotorula sp was considered nonpathogenic, but with the increase of immunosuppressed humans in the last two decades, the species R. mucilaginosa, R. glutinis and R. minuta became potentially pathogenic agents. There have been few veterinary clinical reports associated with Rhodotorula spp. and this work aims to describe the first case of fungal dermatitis caused by R. glutinis in immunosuppressed dog due to visceral leishmaniasis and osteosarcoma in the city of Cuiabá-Mato Grosso. An 11-year-old male mongrel dog was examined to the Veterinary Hospital with skin lesions and the diagnosis was dermatophytosis and the treatment was implemented with topical tea tree oil for one month, but the treatment failed. The animal returned to the Veterinary Hospital with bleeding in the injured area of the tail and biopsy was performed for microbiological and histopathology evaluation. In the microbiological and histopathological analysis, yeast was isolated and yeast-like structures in silver staining were observed, respectively. The isolated yeast was identified as R. glutinis by DNA sequencing. A new treatment was implemented without clinical improvement. The animal was diagnosed with leishmaniasis and osteosarcoma, which probably caused immunosuppression, and its clinical conditions evolved to death.(AU)

Dermatite fúngica associada à Rhodotorula glutinis em cão - Canis familiaris ): relato de caso / Fungal dermatitis associated to Rhodotorula glutinis in dog (Canis familiaris): case report

O fungo Rhodotorula sp. era considerado como não patogênico, mas, com o aumento de casos de humanos imunossuprimidos nas últimas duas décadas, as espécies R. mucilaginosa, R. glutinis e R. minuta se tornaram agentes potencialmente patogênicos. Poucos relatos clínicos veterinários associados à Rhodotorula spp. foram descritos, e o objetivo deste trabalho foi descrever um caso de dermatite fúngica causada por R. glutinis em cão imunossuprimido devido à leishmaniose visceral e osteossarcoma, na cidade de Cuiabá-Mato Grosso. Um cão, macho, 11 anos, sem raça definida, foi trazido ao Hospital Veterinário apresentando lesões de pele com diagnóstico de dermatofitose e foi prescrito tratamento tópico à base de óleo de melaleuca, sem melhora após um mês de tratamento. O animal retornou ao Hospital Veterinário com hemorragia na região lesionada da cauda, e foi realizada biópsia para exames microbiológico e histopatológico. Nas análises microbiológica e histopatológica, foi isolada uma levedura e detectada a presença de estrutura semelhante a leveduras na coloração de prata, respectivamente. A levedura isolada foi identificada como R. glutinis por meio do sequenciamento do DNA. Um novo tratamento foi instituído, sem melhora do quadro clínico. O animal foi diagnosticado também com leishmaniose e osteossarcoma, provavelmente ocasionando a immunossupressão, e seu quadro evoluiu a óbito.(AU)

The fungi Rhodotorula sp was considered nonpathogenic, but with the increase of immunosuppressed humans in the last two decades, the species R. mucilaginosa, R. glutinis and R. minuta became potentially pathogenic agents. There have been few veterinary clinical reports associated with Rhodotorula spp. and this work aims to describe the first case of fungal dermatitis caused by R. glutinis in immunosuppressed dog due to visceral leishmaniasis and osteosarcoma in the city of Cuiabá-Mato Grosso. An 11-year-old male mongrel dog was examined to the Veterinary Hospital with skin lesions and the diagnosis was dermatophytosis and the treatment was implemented with topical tea tree oil for one month, but the treatment failed. The animal returned to the Veterinary Hospital with bleeding in the injured area of the tail and biopsy was performed for microbiological and histopathology evaluation. In the microbiological and histopathological analysis, yeast was isolated and yeast-like structures in silver staining were observed, respectively. The isolated yeast was identified as R. glutinis by DNA sequencing. A new treatment was implemented without clinical improvement. The animal was diagnosed with leishmaniasis and osteosarcoma, which probably caused immunosuppression, and its clinical conditions evolved to death.(AU)

Inhibition of symbiote fungus of the leaf cutter ant Atta sexdens by secondary metabolites from the bacterium Xenorhabdus szentirmaii associated with entomopathogenic nematodes / Inibição do fungo simbionte da formiga-cortadeira Atta sexdens por metabólitos secundários da bactéria Xenorhabdus szentirmaii associada a nematoides entomopatogênicos

Leaf-cutter ants (Hymenoptera: Formicidae) have evolved as dominant herbivores on the American continent. These social insects remove the leaves of economically important plant species to maintain their colony's food reserves, the symbiotic fungus Leucocoprinus gongylophorus, a basidiomycete. Such fungus can be used for applications of fungicide molecules from metabolites generated by symbiont bacteria (Xenorhabdus and Photorhabdus) from entomopathogenic nematodes (Steinernema and Heterorhabditis). Through isolation and multiplication in tryptic soy broth (TSB) medium of the bacteria Xenorhabdus szentirmaii isolated PAM 25, we conducted laboratorial tests using treatments with 10, 25, and 50% of the metabolites obtained in the sixth day of cultivation. The treatments were centrifuged and filtered to generate a supernatant, which was diluted in potato + dextrose + agar (PDA), to verify the consequences of exposure to the fungus L. gongylophorus in Petri dishes. To confirm metabolite efficiency, the control treatments in PDA only and mixed (PDA+TSB) media were conducted simultaneously for 14 days. We observed total inhibition of the symbiont fungus in both the 25 and 50% dilutions during the first days of the tests. Our results support that these metabolites have inhibitory effect on the development of symbiont fungus of leaf-cutter ants.(AU)

As formigas-cortadeiras (Hymenoptera: Formicidae) evoluíram estabelecendo-se no continente americano como herbívoros dominantes. Esses insetos sociais praticam a desfolha de espécies vegetais de interesse econômico, com a finalidade de manter a reserva alimentar da colônia, o fungo simbionte Leucocoprinus gongylophorus, um basidiomiceto. Tal fungo pode ser alvo de aplicações de moléculas fungicidas encontradas em metabólitos gerados por bactérias simbiontes (Xenorhabdus e Photorhabdus) de nematoides entomopatogênicos (Steinernema e Heterorhabditis). Por meio do isolamento e da multiplicação em meio caldo triptona de soja (TSB) da bactéria Xenorhabdus szentirmaii (isolado PAM 25), foram realizados testes laboratoriais com tratamentos contendo 10, 25 e 50% do metabólito obtido no sexto dia de cultivo. Para tanto, o metabólito foi centrifugado e filtrado, gerando um sobrenadante, o qual foi diluído em batata + dextrose + ágar (BDA), para verificar as consequências da exposição do fungo L. gongylophorus em placas de Petri. Os tratamentos de controle apenas com meio BDA e misto (BDA + TSB) também foram conduzidos simultaneamente por 14 dias, a fim de confirmar a eficiência dos metabólitos. Tanto na diluição de 25 e 50% houve total inibição do fungo simbionte já nos primeiros dias da condução dos testes. Diante dos resultados obtidos, pode-se afirmar que os metabólitos são a causa do efeito inibitório do desenvolvimento do fungo simbionte das formigas-cortadeiras.(AU)

Inhibition of symbiote fungus of the leaf cutter ant Atta sexdens by secondary metabolites from the bacterium Xenorhabdus szentirmaii associated with entomopathogenic nematodes / Inibição do fungo simbionte da formiga-cortadeira Atta sexdens por metabólitos secundários da bactéria Xenorhabdus szentirmaii associada a nematoides entomopatogênicos

Leaf-cutter ants (Hymenoptera: Formicidae) have evolved as dominant herbivores on the American continent. These social insects remove the leaves of economically important plant species to maintain their colony's food reserves, the symbiotic fungus Leucocoprinus gongylophorus, a basidiomycete. Such fungus can be used for applications of fungicide molecules from metabolites generated by symbiont bacteria (Xenorhabdus and Photorhabdus) from entomopathogenic nematodes (Steinernema and Heterorhabditis). Through isolation and multiplication in tryptic soy broth (TSB) medium of the bacteria Xenorhabdus szentirmaii isolated PAM 25, we conducted laboratorial tests using treatments with 10, 25, and 50% of the metabolites obtained in the sixth day of cultivation. The treatments were centrifuged and filtered to generate a supernatant, which was diluted in potato + dextrose + agar (PDA), to verify the consequences of exposure to the fungus L. gongylophorus in Petri dishes. To confirm metabolite efficiency, the control treatments in PDA only and mixed (PDA+TSB) media were conducted simultaneously for 14 days. We observed total inhibition of the symbiont fungus in both the 25 and 50% dilutions during the first days of the tests. Our results support that these metabolites have inhibitory effect on the development of symbiont fungus of leaf-cutter ants.(AU)

As formigas-cortadeiras (Hymenoptera: Formicidae) evoluíram estabelecendo-se no continente americano como herbívoros dominantes. Esses insetos sociais praticam a desfolha de espécies vegetais de interesse econômico, com a finalidade de manter a reserva alimentar da colônia, o fungo simbionte Leucocoprinus gongylophorus, um basidiomiceto. Tal fungo pode ser alvo de aplicações de moléculas fungicidas encontradas em metabólitos gerados por bactérias simbiontes (Xenorhabdus e Photorhabdus) de nematoides entomopatogênicos (Steinernema e Heterorhabditis). Por meio do isolamento e da multiplicação em meio caldo triptona de soja (TSB) da bactéria Xenorhabdus szentirmaii (isolado PAM 25), foram realizados testes laboratoriais com tratamentos contendo 10, 25 e 50% do metabólito obtido no sexto dia de cultivo. Para tanto, o metabólito foi centrifugado e filtrado, gerando um sobrenadante, o qual foi diluído em batata + dextrose + ágar (BDA), para verificar as consequências da exposição do fungo L. gongylophorus em placas de Petri. Os tratamentos de controle apenas com meio BDA e misto (BDA + TSB) também foram conduzidos simultaneamente por 14 dias, a fim de confirmar a eficiência dos metabólitos. Tanto na diluição de 25 e 50% houve total inibição do fungo simbionte já nos primeiros dias da condução dos testes. Diante dos resultados obtidos, pode-se afirmar que os metabólitos são a causa do efeito inibitório do desenvolvimento do fungo simbionte das formigas-cortadeiras.(AU)

Ergosterol isolated from the basidiomycete Pleurotus salmoneostramineus affects Trypanosoma cruzi plasma membrane and mitochondria

Abstract Background Major drawbacks of the available treatment against Chagas disease (American trypanosomiasis) include its toxicity and therapeutic inefficiency in the chronic phase of the infection, which makes it a concern among neglected diseases. Therefore, the discovery of alternative drugs for treating chronic Chagas disease requires immediate action. In this work, we evaluated the mushroom Pleurotus salmoneostramineus in the search for potential antiparasitic compounds. Methods Fruit bodies of the basidiomycete Pleurotus salmoneostramineus were triturated and submitted to organic solvent extraction. After liquid-liquid partition of the crude extract, three fractions were obtained and the bioguided fractionation study was conducted to isolate the active metabolites. The elucidation of the chemical structure was performed using GC-MS and NMR techniques. The biological assays for antiparasitic activity were carried out using trypomastigotes of Trypanosoma cruzi and murine macrophages for mammalian cytotoxicity. The mechanism of action of the isolated compound used different fluorescent probes to evaluate the plasma membrane permeability, the potential of the mitochondrial membrane and the intracellular levels of reactive oxygen species (ROS). Results The most abundant fraction showing the antiparasitic activity was isolated and chemically elucidated, confirming the presence of ergosterol. It showed anti-Trypanosoma cruzi activity against trypomastigotes, with an IC50 value of 51.3 g/mL. The compound demonstrated no cytotoxicity against mammalian cells to the maximal tested concentration of 200 g/mL. The mechanism of action of ergosterol in Trypanosoma cruzi trypomastigotes resulted in permeabilization of the plasma membrane, as well as depolarization of mitochondrial membrane potential, leading to parasite death. Nevertheless, no increase in ROS levels could be observed, suggesting damages to plasma membrane rather than an induction of oxidative stress in the parasite. Conclusions The selection of naturally antiparasitic secondary metabolites in basidiomycetes, such as ergosterol, may provide potential scaffolds for drug design studies against neglected diseases.