Environmental drivers of tropical forest snake phenology: Insights from citizen science.

Museum specimens and citizen science initiatives are valuable sources of information on how anthropogenic activities affect biodiversity and how species respond to rapid global change. Although tropical regions harbor most of the planet's biodiversity, investigations on species' phenological changes are heavily biased toward temperate regions. Such unevenness in phenological research is also taxonomically biased, with reptiles being the least studied group among tetrapod species regarding animal phenology. Herein, we used long-term time-series data to investigate environmentally driven changes in the activity pattern of tropical forest snakes. We gathered natural history collection and citizen science data for 25 snake species (five venomous and 20 non-venomous) from an Atlantic Forest region in southeastern Brazil. Using circular mixed-effects models, we investigate whether snake activity patterns followed the variation in environmental variables over a decade. Our results show that the activity pattern of Atlantic Forest snakes was seasonal and largely driven by average temperature and relative humidity. Since snakes are ectothermic animals, they are particularly sensitive to temperature variations, especially at small scales. Moreover, relative humidity can affect snake's seasonal activities through physiological constraints and/or prey availability. Most specimens were registered during the rainy season, with highly venomous snakes (lanceheads and coral snakes) emerging as the most abundant taxa. We highlight the importance of citizen science and natural history collections in better understanding biodiversity. Furthermore, our data obtained from local collectors underscore the need for environmental education programs and collaboration between researchers and local decision-makers to raise awareness and reduce conflicts between people and snakes in the region.

Seasonal and habitat structure of an anuran assemblage in a semideciduous forest area in Southeast Brazil.

In this study, we evaluated the reproductive activity and the temporal and spatial distributions of anuran assemblages in three environments within a semideciduous forest in Southeast Brazil, located at Municipality of Barão de Monte Alto, State of Minas Gerais, Brazil. The field activities were carried out during three consecutive days, monthly throughout the rainy seasons of 2013-2014 and 2014-2015. We recorded 28 anurans species, distributed in eight families. We observed the spatial-temporal distribution of some species, and their associated reproductive behaviors through exploration of vocalizations at different sites. The spatial and temporal distribution of the species seems to adapt to abiotic and biotic factors of their environment.

Cytogenetics of Four Species of the Green Clade Aplastodiscus Lutz, 1950 (Anura: Cophomantinae): New Insights into the Chromosomal Evolution of the Genus.

The tree frog Aplastodiscus is a Neotropical taxon that encompasses 15 species in the Atlantic forest biome, with one isolated species in the Central Brazilian Cerrado. To date, only 8 species have been karyotyped, showing high levels of diploid number variation, which allowed clustering species in chromosome number groups: 2n = 24 (Aplastodiscus perviridis group), 2n = 22 (Aplastodiscus albofrenatus group), 2n = 20, and 2n = 18 (both within Aplastodiscus albosignatus group). This study aims to report karyotypic information on 4 species from the last 2 groups using classical and molecular cytogenetic techniques and hypothesize chromosomal evolutionary trends within the species groups. Aplastodiscus weygoldti showed 2n = 22; Ag-NOR and FISH 18S rDNA signals were located in the interstitial region of the short arms of chromosome pair 6. Aplastodiscus cavicola, Aplastodiscus sp. 4, and Aplastodiscus sp. 6 showed 2n = 18; Ag-NOR and FISH 18S rDNA bands were located in the terminal region of the long arm of chromosome pair 9. Our results support multiple and independent chromosome fusion events within Aplastodiscus, including a new chromosome fission event. Ag-NOR and FISH 18S rDNA patterns were restricted to the small chromosome pairs, similar to the other species within this genus, and confirm overall chromosome morphology conservation among the genera of Cophomantinae.

Distribution pattern of anurans from three mountain complexes in southeastern Brazil and their conservation implications.

Biogeographic tools support spatial distribution pattern hypotheses and help to determine priority areas for conservation. Our aim was to verify biogeographic patterns for anurans in three mountain complexes in southeastern Brazil, as well as to discuss the status of species conservation recorded and the biogeographical units evaluated. We selected 16 areas distributed in the Serra da Mantiqueira complex, south of Serra do Espinhaço and Serra da Canastra. We used the occurrence (geographic coordinates) of each species in the localities to determine areas of endemism applying the Endemicity Analysis method. We also tested whether similarity between areas was explained by geographic distance (Multiple Regression on distance Matrices-MRM). The Serra do Itatiaia, Serra da Canastra, Plateau of Poços de Caldas and Serra do Cipó were the areas that presented the highest number of species restricted to them. Through the Endemicity Analysis, we identified four areas of endemism with higher scores. The MRM revealed that the geographic distance explained 41% of species dissimilarity between areas. Most of the endemic species from these areas have inaccurate conservation statuses (data deficient or unevaluated). These results highlight the need for greater research efforts towards understanding species restricted by distribution, as well as the priority in conserving these endemic areas.

Molecular phylogeny of Neotropical rock frogs reveals a long history of vicariant diversification in the Atlantic forest.

The Brazilian Atlantic coastal forest is one of the most heterogeneous morphoclimatic domains on earth and is thus an excellent region in which to examine the role that habitat heterogeneity plays in shaping diversification of lineages and species. Here we present a molecular phylogeny of the rock frogs of the genus Thoropa Cope, 1865, native to the Atlantic forest and extending to adjacent campo rupestre of Brazil. The goal of this study is to reconstruct the evolutionary history of the genus using multilocus molecular phylogenetic analyses. Our topology reveals 12 highly supported lineages among the four nominal species included in the study. Species T. saxatilis and T. megatympanum are monophyletic. Thoropa taophora is also monophyletic, but nested within T. miliaris. Populations of T. miliaris cluster in five geographically distinct lineages, with low support for relationships among them. Although all 12 lineages are geographically structured, some T. miliaris lineages have syntopic distributions with others, likely reflecting a secondary contact zone between divergent lineages. We discuss a biogeographic scenario that best explains the order of divergence and the distribution of species in Atlantic forest and adjacent areas, and outline the implications of our findings for the taxonomy of Thoropa.

The antimicrobial peptide HS-1 inhibits dengue virus infection.

Dengue virus (DENV) is an arbovirus that belongs to the Flaviviridae family. Studies reveal that peptides secreted by amphibians have many functions, such as antiviral and antimicrobial activities. As there is no antiviral drug effective against the DENV, the antiviral activity of a synthetic peptide called HS-1, derived from the secretion of the anuran Hypsiboas semilineatus, has been evaluated. The assays of neutralization in the Vero cells show a complete inhibition of infection of the serotypes 2 and 3. Furthermore, the direct action of peptides on the viral particle can be observed through atomic force microscopy. In vivo tests display 80% protection against the dengue-2 virus due to the presence of HS-1, which reveals its potential as an antiviral against the DENV.

Redescription of the Advertisement Call of Five Species of Thoropa (Anura, Cycloramphidae), Including Recordings of Rare and Endangered Species.

Frogs of the genus Thoropa comprise six endemic Brazilian species on the Eastern side of the country. Little is known about their natural history, especially about their acoustic communication. Therefore, aiming to provide an overview of their vocalizations, we analyzed and redescribed male advertisement calls of three living and two possibly extinct species. The smaller species, T. petropolitana and T. lutzi, produce simple calls (one single note) with a higher frequency range than the remaining larger ones. On the other hand, the larger species present complex calls, with more than one note: T. megatympanum calls have three notes, T. taophora calls have four notes, and T. miliaris calls varies from three to six notes. Population snout-vent length negatively correlated with peak of dominant frequency as expected. However, highlighted differences between two populations of T. lutzi, which could indicate need of further taxonomic evaluation of those lineages. Peculiar morphology, such as the absence of vocal sacs and slits, may have contributed to their call variation and highly banded frequency structure. If the observed population differences reflect species-level differences, T. lutzi may be classified as a critically endangered species, as T. petropolitana. Furthermore, we provided a suggestion to an unusual behavior in frogs: calling with the mouth open in the smaller species of the genus.

Karyological study of Ololygon tripui (Lourenço, Nascimento and Pires, 2009), (Anura, Hylidae) with comments on chromosomal traits among populations.

To increase the number of cytogenetic characters used in Ololygon tripui systematics, we applied some cytogenetic techniques such as Giemsa, C- and NOR-banding, and fluorescence in situ hybridization (FISH) with 18S rDNA and repetitive microsatellite DNA probes to the study of four populations from Minas Gerais State (southeastern Brazil). All populations showed 2n = 24 and FN = 48, and chromosomal formula 8m + 10sm + 6st. Nucleolar organizing regions (NORs) were located on chromosome pair 6 in all populations, although in the Tripuí locality additional markings were observed on one homologue of chromosome pair 3. These patterns were partially congruent with results obtained using the 18S rDNA FISH probe. The microsatellites repetitive DNA (GA)15 and (CAT)10 probes accumulated predominantly in the terminal region of all chromosomes. Chromosome morphology and Ag-NOR were conserved among populations, a conserved pattern in Ololygon Fitzinger, 1843. Repetitive DNA FISH probes patterns were similar among populations, but they revealed species-specific differences when compared with other species of the genus Ololygon, suggesting that molecular cytogenetics are potentially more informative in karyologically conservative taxa.

On the diagnosis and conservation of the poorly known bromeligenous Scinax arduous Peixoto, 2002 (Amphibia; Anura; Hylidae).

Since Scinax arduous description, many other populations belonging to the Scinax perpusillus group have been recorded for the States of Espírito Santo and Minas Gerais, Southeastern Brazil. Both in collections and publications most of these new specimens are identified as S. arduous, Scinax cf. arduous, Scinax cf. perpusillus, Scinax gr. perpusillus, S. perpusillus and S. v-signatus. Such state of affairs may be due to the lack of information on the original description of S. arduous. Only two individuals [the holotype (female) and the paratype (male)] were used in the original description and diagnosis, therefore, information on variation and distribution were not available. Furthermore, in S. arduous description, the section on coloration in life was based only on two juveniles raised in laboratory. Herein we redefine Scinax arduous based on 44 males and 17 females from the type locality, the Municipality of Santa Teresa, in the State of Espírito Santo. In addition, we provide information on its conservation status, distribution, natural history, vocalization, tadpoles, and detailed photographs of both adults and larvae.

Identification and characterization of an antimicrobial peptide of Hypsiboas semilineatus (Spix, 1824) (Amphibia, Hylidae).

The multidrug-resistant bacteria have become a serious problem to public health. In this scenery the antimicrobial peptides (AMPs) derived from animals and plants emerge as a novel therapeutic modality, substituting or in addition to the conventional antimicrobial. The anurans are one of the richest natural sources of AMPs. In this work several cycles of cDNA cloning of the skin of the Brazilian treefrog Hypsiboas semilineatus led to isolation of a precursor sequence that encodes a new AMP. The sequence comprises a 27 residue signal peptide, followed by an acidic intervening sequence that ends in the mature peptide at the carboxy terminal. The AMP, named Hs-1, has 20 amino acids residues, mostly arranged in an alpha helix and with a molecular weight of 2144.6 Da. The chemically synthesized Hs-1 showed an antimicrobial activity against all Gram-positive bacteria tested, with a range of 11-46 µM, but it did not show any effect against Gram-negative bacteria, which suggest that Hs-1 may have a selective action for Gram-positive bacteria. The effects of Hs-1 on bacterial cells were also demonstrated by transmission electron microscopy. Hs-1 is the first AMP to be described from H. semilineatus.

Anurans in a forest remnant in the transition zone between cerrado and Atlantic Rain Forest domains in Southeastern Brazil.

This study presents the species richness, temporal distribution and reproductive activity of anurans from the Uaimií State Forest (Floresta Estadual do Uaimií - FLOE Uaimií), situated in the Quadrilátero Ferrífero region, municipality of Ouro Preto, Minas Gerais state, Brazil. Field activities were performed monthly from September 2009 to August 2010. We recorded 36 anurans species, distributed in 10 families. The greatest richness of the sampled sites corresponds to a permanent rivulet in a secondary forest. The majority of anuran species presented seasonal vocalization activity pattern, mainly in the rainy season. The anuran species composition of FLOE Uaimií is similar to others studied areas from the Quadrilátero Ferrífero region.