Unravelling the fungal darkness in a tropical cave: richness and the description of one new genus and six new species.

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.

Diagnosis of canine visceral leishmaniasis: biotechnological advances.

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.

Canine visceral leishmaniosis: a comparative analysis of the EIE-leishmaniose-visceral-canina-Bio-Manguinhos and the IFI-leishmaniose-visceral-canina-Bio-Manguinhos kits.

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.

Description of semen characteristics from six-banded armadillos (Euphractus sexcinctus) collected by electroejaculation.

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.

Níveis séricos da gama-glutamiltransferase em cães com e sem infecção natural por Leishmania (Leishmania) chagasi / Serum levels of gamma-glutamyl transferase in dogs naturally infected or not by Leishmania (Leishmania) Chagasi

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.