RESUMO: Estudos sobre a microbiota fúngica de répteis e anfíbios são necessários para melhor compreender as interações hospedeiro-microrganismo e o estabelecimento de doenças fúngicas nesses animais. No entanto, esses estudos são limitados. O objetivo da presente pesquisa foi identificar leveduras isoladas de répteis e anfíbios do bioma Caatinga e avaliar a produção de fatores de virulência, a sensibilidade a antifúngicos no crescimento planctônico e de biofilme e a patogenicidade de Candida famata. Vinte e nove isolados dos gêneros Candida, Cryptococcus e Rhodotorula foram identificados por métodos fenotípicos e/ou moleculares e a produção de enzimas hidrolíticas in vitro por esses gêneros de fungos foi avaliada. Além disso, foi avaliada a suscetibilidade de células planctônicas e biofilmes a azólicos e anfotericina B. A patogenicidade de C. famata, a espécie de levedura isolada mais prevalente, foi avaliada usando Caenorhabditis elegans. C. famata foi a levedura mais prevalente na microbiota de anfíbios e répteis. A produção de fosfolipase e protease foi observada em 18/29 e 11/29 dos isolados de levedura, respectivamente, enquanto 100% formaram biofilmes. O itraconazol apresentou altas concentrações inibitórias mínimas contra C. famata e C. tropicalis. A anfotericina B reduziu a biomassa e atividade metabólica dos biofilmes. C. famata induziu a mortalidade de C. elegans. Em conclusão, répteis e anfíbios são colonizados por leveduras capazes de produzir importantes fatores de virulência, especialmente por cepas de Candida spp. que apresentam baixa suscetibilidade a azólicos que podem resultar de desequilíbrio no ecossistema. Por fim, C. famata isolados desses animais apresentaram alta patogenicidade, mostrando a importância do estudo da microbiota fúngica de répteis e anfíbios.

ABSTRACT: Studies on the fungal microbiota of reptiles and amphibians are necessary to better understand of host-microbe interactions and the establishment of fungal disease in these animals. However, these studies are limited. The present researchidentified yeasts from free-ranging reptiles and amphibians from the Caatinga biome andevaluated the virulence factors production, the antifungal susceptibility in planktonic and biofilm growth and the pathogenicity of Candida famata isolates. Twenty-nine isolates of the genera Candida, Cryptococcus and Rhodotorula were identified by phenotypic and/or molecular methods and production of hydrolytic enzymes in vitro by these genera of fungi was evaluated. In addition, susceptibility of planktonic cells and biofilms to azoles and amphotericin B was evaluated. The pathogenicity of C. famata, the most prevalent yeast species isolated, was evaluated using Caenorhabditis elegans model. C. famata was the most prevalent yeast in amphibian and reptilian microbiota. Phospholipase and protease production was observed in 18/29 and 11/29 of the yeast isolates, respectively, while 100% formed biofilms. Itraconazole presented high minimal inhibitory concentrations against C. famata and C. tropicalis. Amphotericin B reduced the biomass and metabolic activity of biofilms. C. famata induced the mortality of C. elegans. In conclusion, reptiles and amphibians are colonized by yeasts capable of producing important virulence factors, especially by Candida spp. that present low susceptibility to azoles which may result from imbalances in ecosystem. Finally, C. famata isolated from these animals presented high pathogenicity, showing the importance of the study of reptile and amphibians fungal microbiota.

ABSTRACT A list of spider species is presented for the Belém Area of Endemism, the most threatened region in the Amazon Basin, comprising portions of eastern State of Pará and western State of Maranhão, Brazil. The data are based both on records from the taxonomic and biodiversity survey literature and on scientific collection databases. A total of 319 identified species were recorded, with 318 occurring in Pará and only 22 in Maranhão. About 80% of species are recorded at the vicinities of the city of Belém, indicating that sampling effort have been strongly biased. To identify potentially high-diversity areas, discounting the effect of variations in sampling effort, the residues of a linear regression between the number of records and number of species mapped in each 0.25°grid cells were analyzed. One grid, representing the Alto Turiaçu Indigenous land, had the highest deviation from the expected from the linear regression, indicating high expected species richness. Several other grid cells showed intermediate values of the regression residuals, indicating species richness moderately above to the expected from the model.

O aumento da incidência das infecções fúngicas, bem como o registro crescente de resistência e falha terapêutica, têm impulsionado a realização de estudos de prospecção de fitoquímicos com propriedades antifúngicas. Diante do exposto, o presente estudo investigou o potencial antifúngico de extratos de Baccharis ligustrina, B. schultzii, Croton jacobinensis, Licania rigida, Moringa oleifera, Vernonia sp. e V. brasiliana, e de óleos essenciais de Lippia alba (Quimiotipos 1, 2, 3 e 4) e Ocimum gratissimum. Inicialmente, foi realizada uma avaliação qualitativa da atividade antifúngica de cada amostra por meio do método de difusão em ágar, frente a cepas de Candida albicans e Microsporum canis, mostrando que apenas os extratos de M. oleifera (MLF-C) e Vernonia sp. (TVS-H) apresentaram atividade frente a C. albicans e M. canis, com halos de inibição =10mm. Também foram determinadas a concentração inibitória mínima (CIM), frente a 12 cepas de C. albicans e M. canis, e a toxicidade aguda de MLF-C e TVS-H, através de protocolos do Clinical and Laboratory Standards Institute (CLSI) e ensaio com Artemia sp., respectivamente. A CIM (80%) de MLF-C e TVS-H variou de 0,156 a 2,5mg mL-1 frente C. albicans e de 0,039 a 1,25 e 0,039 a 0,625mg mL-1 para M. canis, respectivamente. A CIM (100%) de MLF-C e TVS-H variou de 0,625 a 2,5mg mL-1 frente C. albicans é de 0,039 a 2,5 e 0,078 a 1,25mg mL-1 para M. canis, respectivamente. As doses letais (DL50) para o MLF-C e TVS-H foram de 201,09 e 204,17mg mL-1, respectivamente, sendo, portanto, demonstrada a baixa toxicidade desses extratos. Os extratos de M. oileifera e Vernonia sp. apresentaram atividade antifúngica frente cepas de C. albicans e M. canis, abrindo a perspectiva de estudos para caracterização dos seus componentes bioativos.

The increase in the incidence of fungal infections and the frequent report of resistance and therapeutic failure has promoted the performance of phytochemical screening for compounds with antifungal properties. Based on this, the present study investigated the antifungal potential of extracts of Baccharis ligustrina, B. schultzii, Croton jacobinensis, Licania rigida, Moringa oleifera, Vernonia sp. and V. brasiliana and of essential oils of Lippia alba (Chemotypes 1, 2, 3 and 4) and Ocimum gratissimum. Initially, a qualitative evaluation of the antifungal activity of each vegetal sample against strains of Candida albicans and Microsporum canis, through the agar diffusion method, was performed. Extracts of M. oleifera (MLF-C) and Vernonia sp. (TVS-H) presented activity against C. albicans and M. canis with inhibition halos =10mm. Then, minimum inhibitory concentrations (MICs) for MLF-C and TVS-H against 12 strains of C. albicans and M. canis were determined through the methodology established by the Clinical and Laboratory Standards Institute (CLSI), and acute toxicity tests against Artemia sp. were performed for both extracts. MICs (80%) for MLF-C and TVS-H varied from 0.156 to 2.5mg mL-1 against C. albicans and from 0.039 to 1.25mg mL-1 and 0.039 to 0.625mg mL-1 against M. canis, respectively. MICs (100%) for MLF-C and TVS-H varied from 0.625 to 2.5mg mL-1 for C. albicans and from 0.039 to 2.5mg mL-1 and 0.078 to 1.25mg mL-1 against M. canis, respectively. Lethal doses (DL50) of MLF-C and TVS-H were 201.09 and 204.17mg mL-1, respectively, being, therefore, demonstrated the low toxicity of these extracts. M. oleifera and Vernonia sp. extracts presented in vitro antifungal activity against C. albicans and M. canis, creating perspectives for the development of studies on the characterizations of their bioactive components.