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Caves are special environments that harbour an incredible diversity of life, including fungal species. Brazilian caves have been demonstrated to be biodiversity hotspots for known and unknown fungal species. We investigated the richness of culturable fungi in a tropical cave in Brazil by isolating these microorganisms from the sediment and air. The fungal abundance of colony-forming units (CFUs) was 3 178 in sediment and 526 in air. We used morphological features and phylogenetic analyses of actin (actA), calmodulin (cmdA), internal transcribed spacer regions and intervening 5.8S rRNA (ITS), large subunit (LSU) rDNA, RNA polymerase II second largest subunit (rpb2), translation elongation factor 1-alpha (tef1), and ß-tubulin (tub2) genes to identify these isolates. Forty-one species belonging to 17 genera of Ascomycota and two of Basidiomycota were identified, and the genus Aspergillus was most commonly observed in the cave (13 taxa). Twenty-four species were found in sediment (16 exclusives) and 25 species were found in air (17 exclusives). In this study, we introduced a new genus (Pseudolecanicillium gen. nov.) in the family Cordycipitaceae and six new species (14 % of the total taxa identified) of fungal isolates obtained from sediment and air: Aspergillus lebretii sp. nov., Malbranchea cavernosa sp. nov., Pseudohumicola cecavii sp. nov., Pseudolecanicillium caatingaense sp. nov., Talaromyces cavernicola sp. nov., and Tritirachium brasiliense sp. nov. In addition, we built a checklist of the fungal taxa reported from Brazilian caves. Our results highlight the contribution of Brazilian caves to the estimation of national and global fungal diversity. Citation: Alves VCS, Lira RA, Lima JMS, Barbosa RN, Bento DM, Barbier E, Bernard E, Souza-Motta CM, Bezerra JDP (2022). Unravelling the fungal darkness in a tropical cave: richness and the description of one new genus and six new species. Fungal Systematics and Evolution 10: 139-167. doi: 10.3114/fuse.2022.10.06.

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The aim of this study is to describe the characteristics of the semen from six-banded armadillos (Euphractus sexcinctus) collected by electroejaculation. Six mature males were physically restrained and electroejaculated twice for the collection of semen. Semen collected was immediately evaluated for appearance, volume, pH, sperm motility, vigor, morphology, percentage of live sperm and functional membrane integrity by light microscopy. Semen was obtained from all (100%) twelve attempts conducted for electroejaculation. Armadillos' semen had a white-translucent appearance, and great viscosity. Mean values obtained in analysis of the semen were: 353+/-86 microl for volume, 9 for pH, 45+/-14 x 10(6)sperm/ml for concentration, 61+/-7% motile sperm with 2+/-0.2 for vigor, 55+/-7% live sperm, 86+/-2% morphologic normal sperm, and 46+/-6% functional membrane integrity. In conclusion, semen from six-banded armadillos can be efficiently obtained by electroejaculation. The characteristics of semen collected by electroejaculation in six-banded armadillos provide background information that may be useful for assisted breeding programs in the members of the Xenarthra family.

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The serum levels of Gamma-GT in dogs naturally infected by Leishmania (Leishmania) chagasi were evaluated. Two groups of animals were used: the first with 41 naturally infect dogs and the second with 14 uninfect animals. The results showed 17.1% (7/41) dogs presented high levels of gamma-GT, but no difference between groups was observed. So, the serum level of gamma-GT is not a tool for diagnosis of visceral leishmaniasis in dogs.

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The serum levels of Gamma-GT in dogs naturally infected by Leishmania (Leishmania) chagasi were evaluated. Two groups of animals were used: the first with 41 naturally infect dogs and the second with 14 uninfect animals. The results showed 17.1% (7/41) dogs presented high levels of gamma-GT, but no difference between groups was observed. So, the serum level of gamma-GT is not a tool for diagnosis of visceral leishmaniasis in dogs.(AU)

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Human visceral leishmaniasis (HVL) is endemic in the tropical and sub-tropical regions of Africa, Asia, the Mediterranean, Southern Europe and South and Central America, with approximately 500,000 new cases reported annually. As dogs are considered to be the major reservoirs for HVL, the accurate diagnosis of disease in these animals is important. Diagnosis of canine visceral leishmaniasis (CVL) is performed mainly by direct parasitological methods that can yield false-negative results, either because of the very low number of Leishmania spp. organisms in clinical samples (bone marrow and lymph nodes) or because morphological identification is difficult. In addition, these methods are invasive. Conventional serological techniques are limited by cross-reactivity with other parasitic diseases and because several technical procedures have not been standardised. The development of polymerase chain reaction based approaches and immunoassays based on the use of recombinant antigens aimed at improving the sensitivity and specificity of CVL diagnosis is discussed.

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ABSTRACT Mascagnia rigida, a vine or shrub from the Malpighiaceae family popularly named in Brazil as tingui and timbó, is one of most well-known poisonous plants from Northeast and part of Southwest Brazil. It represents a significant cause of death in cattle and goats. In light of the lack of any previous published report of any case of its natural poisoning in sheep, the present article reports such poisoning for the first time, describing four outbreaks of natural poisoning by M. rigida in sheep. The outbreaks occurred in the counties of Mossoró (3 outbreaks) and Upanema (1 outbreak), state of Rio Grande do Norte, Brazil, in 2006. Clinical manifestations in affected animals consisted of apathy, muscle trembling, tachycardia, difficulty in standing up, dyspnea, convulsions, and death. Histological lesions were multifocal infiltrate of lymphocytes in the myocardium associated with edema and congestion of myocytes, and diffuse vacuolar degeneration in hepatocytes. In order to confirm the etiology of the disease, an experiment was performed with two female healthy sheep that received daily fresh leaves of M. rigida. On the first and second days the plant was administered at a dose of 10 g/kg body weight, whereas on the third and fourth days the dose was 20 g/kg. About 2 hours after the last dosing (fourth day) the animals presented clinical disturbances similar to natural poisoning, including death. Pathological findings were also similar to natural cases. Thus, sheep are also naturally affected by M. rigida poisoning, and there is a cumulative effect, the total lethal dose being 60 g/kg of body weight.

RESUMO Mascagnia rigida, cipó ou arbusto da família Malpighiaceae, denominada popularmente como tingui e timbó, é uma das plantas tóxicas mais conhecidas, presente na região Nordeste e parte da região Sudeste do Brasil. Representa uma significante causa de morte em bovinos e caprinos. No entanto, até o momento, não foi descrito nenhum caso de intoxicação natural em ovinos. Assim, o presente trabalho procura descrever, pela primeira vez, quatro surtos de intoxicação por M. rigida em ovinos. Os surtos ocorreram nos Municípios de Mossoró (três surtos) e Upanema (um surto), no Rio Grande do Norte, em 2006. A sintomatologia clínica dos animais afetados consistiu em apatia, tremores musculares, taquicardia, dificuldade em se manter em pé, dispnéia e convulsões seguidas de morte. As lesões histológicas observadas foram infiltrado multifocal de células linfocitárias no miocárdio associado a edema e degeneração dos miócitos, e degeneração vacuolar difusa dos hepatócitos. Para confirmar a etiologia da doença, foi realizado um experimento com duas ovelhas saudáveis administrando-se diariamente folhas frescas da M. rigida. No primeiro e no segundo dia foi administrada a planta na dose de 10 g/kg de peso vivo, enquanto no terceiro e no quarto dia, a dose foi de 20 g/kg. Aproximadamente 2 horas após a última administração (quarto dia), os animais apresentaram manifestações clínicas semelhantes à intoxicação natural, inclusive morte. Os achados patológicos também foram similares aos casos naturais. Assim, a espécie ovina também é naturalmente afetada pela intoxicação por M. rigida e há um efeito cumulativo, sendo a dose letal total de 60 g/kg de peso vivo.

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This study evaluated the performance of the EIE-leishmaniose-visceral-canina-Bio-Manguinhos (EIE-LVC) kit and to compare it with that of the IFI-leishmaniose-visceral-canina-Bio-Manguinhos (IFI-LVC) kit. Four groups of dogs were studied: group 1 (G1), dogs with clinical signs indicative of CVL and testing positive for the parasite (n = 25); group 2 (G2), dogs with only a presumed diagnosis of CVL (n = 62); group 3 (G3), dogs that had never lived in an area where CVL is endemic and never received a blood transfusion (n = 16); group 4 (G4), dogs carrying other parasites: such as babesiosis (n = 4), ehrlichiosis (n = 6) and demodicosis (n = 1). G1 and G3 were used for the calculation of sensitivity and specificity, respectively. The EIE-LVC showed a sensitivity of 72% (IC 95%: 50.4-87.1%) and a specificity of 87.5% (IC 95%: 60.4-97.8%). The value of the kappa index was 0.975 (CI 95%: 0.926-1.024), which represents an excellent fit. For IFI-LVC, the sensitivity was 68.0% (CI 95%: 46.4-84.3%) and the specificity 87.5% (CI 95%: 60.4-97.8%). When the tests were conducted in parallel, sensitivity was 92.0% (CI 95%: 72.5-98.6%) and specificity 75.0% (CI 95%: 47.4-91.7%). However, when conducted consecutively, the tests showed a sensitivity of 48.0% (CI 95%: 28.3-68.2%) and a specificity of 100.0% (CI 95%: 75.9-99.4%). The analysis of clinically suspected dogs using IFI-LVC and EIE-LVC kits in parallel, revealed that 26/62 animals were positive. Cross-reaction was observed in a dog with demodicosis. These results lead to the following conclusions: (1) the performance of the EIE-LVC kit is not statistically different from the IFI-LVC and (2) the kits must be used in parallel if higher sensitivity is required, reducing the number of false-negative results.

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