Sympatric and independently evolving lineages in the Thoropa miliaris - T. taophora species complex (Anura: Cycloramphidae).

Species delimitation can be challenging and affected by subjectivity. Sibling lineages that occur in sympatry constitute good candidates for species delimitation regardless of the adopted species concept. The Thoropa miliaris + T. taophora species complex exhibits high genetic diversity distributed in several lineages that occur sympatrically in the southeastern Atlantic Forest of Brazil. We used 414 loci obtained by anchored hybrid enrichment to characterize genetic variation in the Thoropa miliaris species group (T. saxatilis, T megatympanum, T. miliaris, and T. taophora), combining assignment analyses with traditional and coalescent phylogeny reconstruction. We also investigated evolutionary independence in co-occurring lineages by estimating gene flow, and validated lineages under the multispecies coalescent. We recovered most previously described lineages as unique populations in assignment analyses; exceptions include two lineages within T. miliaris that are further substructured, and the merging of all T. taophora lineages. We found very low probabilities of gene flow between sympatric lineages, suggesting independent evolution. Species tree inferences and species delimitation yielded resolved relationships and indicate that all lineages constitute putative species that diverged during the Pliocene and Pleistocene, later than previously estimated.

Does reproductive effort influence the metabolic rate of Tomodon dorsatus snakes?

In Squamate reptiles, vitellogenesis in females is marked by an increase in the number and size of follicles, leading to an increase in aerobic metabolism to support the rising energy demand during reproductive activity. Male gametes are smaller than female gametes, and spermiogenesis requires less energy than vitellogenesis. This investigation compares the relationship between aerobic metabolism and reproductive effort in male and female viviparous Tomodon dorsatus snakes. We evaluated the influence of the males' and females' reproductive stages, along with the masses of follicles and testicles, on their aerobic metabolic rate. Our findings show that females have higher maintenance cost of pregnancy than metabolic costs of vitellogenesis. In males, we found no evidence of the reproductive effort influencing aerobic metabolic rates. In T. dorsatus, females have a long period of development and high fecundity. Therefore, the metabolic costs of vitellogenesis represent a critical component in the reproductive effort. On the other hand, males of T. dorsatus maintain a similar mass of gonads throughout the year with constant energy demand. These results reflect the difference in reproductive efforts between females and males of T. dorsatus.

Herpetofauna from an Atlantic Forest fragment in São Paulo, Brazil

Habitat loss via urbanization is a cause of decline for reptile and amphibian species around the world. The Atlantic Forest is the second largest rainforest of South America and holds one of the most extraordinary herpetofauna in the world, even after decades of fragmentation. The Parque Estadual das Fontes do Ipiranga (PEFI) is one of the most important remnants of Atlantic Forest inside the city of São Paulo (Brazil), but its herpetofauna richness has not been studied. Between 2011 and 2014, we carried out a herpetofaunal inventory in the PEFI and made notes on habitat use by using pitfall traps with drift fences combined with acoustic and visual encounter surveys. We also collected data from opportunistic encounters and historical information from scientific collections. We found that the PEFI harbors 22 native species of anurans, 24 native species of reptiles, and three exotic species of reptiles. Also, through surveying scientific collections, we found three species of amphibians and one of the snakes that were detected in the past but were not detected during our survey. The composition of amphibian species in the PEFI is similar to proximal small urban and periurban Atlantic Forest fragments, but its species richness is higher than these fragments even when compared to areas with similar or larger size. In conclusion, the highly urban PEFI supports considerable herpetofaunal diversity in comparison with other regional areas and our data reinforce the necessity of conserving this and other urban forest fragments through strategic management across the Atlantic Forest.

Does reproductive effort influence the metabolic rate of Tomodon dorsatus snakes?

In Squamate reptiles, vitellogenesis in females is marked by an increase in the number and size of follicles, leading to an increase in aerobic metabolism to support the rising energy demand during reproductive activity. Male gametes are smaller than female gametes, and spermiogenesis requires less energy than vitellogenesis. This investigation compares the relationship between aerobic metabolism and reproductive effort in male and female viviparous Tomodon dorsatus snakes. We evaluated the influence of the males' and females' reproductive stages, along with the masses of follicles and testicles, on their aerobic metabolic rate. Our findings show that females have higher maintenance cost of pregnancy than metabolic costs of vitellogenesis. In males, we found no evidence of the reproductive effort influencing aerobic metabolic rates. In T. dorsatus, females have a long period of development and high fecundity. Therefore, the metabolic costs of vitellogenesis represent a critical component in the reproductive effort. On the other hand, males of T. dorsatus maintain a similar mass of gonads throughout the year with constant energy demand. These results reflect the difference in reproductive efforts between females and males of T. dorsatus.

Unexpected reproductive fidelity in a polygynous frog.

Polygynous mating systems with group fidelity are a common animal organization, typically consisting of multiple females in a mated group with a single male for an extended period (sometimes referred to as harem polygyny). Single-male polygyny with reproductive fidelity occurs in invertebrates, bony fishes, and some tetrapods, such as lizards, mammals, and birds. In amphibians, reproductive fidelity in polygynous groups is not fully demonstrated. Combining data on larval development, molecular paternity assignment, and in situ behavioral observations, we reveal high fidelity during a prolonged breeding season in a Neotropical polygynous frog. Males dominate scarce breeding sites, guarding offspring, and mating exclusively with multiple females that exhibit dominance rank. This system likely evolved in response to intense competition for breeding sites and intrasexual competition for mates.

Thermal and salinity effects on locomotor performance of Thoropa taophora tadpoles (Anura, Cycloramphidae).

It is well known that environmental temperature influences several biological functions of ectotherms, notably in amphibians. The high permeability of anuran skin, associated with the effect of elevated environmental temperature, potentiates the dehydration process and this combination may restrict locomotor performance. Thoropa taophora is an endemic species from the Atlantic Rainforest whose tadpoles are semiterrestrial and predominantly diurnal, and are found in rocky seashores where they are exposed to sea spray and high temperatures. In this study we investigated how temperature and salinity conditions affect the locomotor performance in Thoropa taophora tadpoles. We also assessed how different osmotic concentrations affect the activity of the metabolic pathways that support muscle function. We measured the sprint speed of tadpoles of various sizes at different temperatures and salinities in the field. We also measured the activity of the enzymes pyruvate kinase (PK), lactate dehydrogenase (LDH) and citrate synthase (CS) in different temperatures and osmotic concentrations, and calculated the thermal sensitivity and the activity constants for each osmolality. Our results showed that, in general, sprint speed decreased with increasing temperature and salinity. However, whereas the effect of increased salinity was similar in smaller and larger tadpoles, increased temperature had a higher negative impact on sprint speed of larger tadpoles, thus indicating low thermal sensitivity of small tadpoles. PK and LDH thermal sensitivities and LDH constant of activity decreased as the osmolality increased. In conclusion, the locomotor capacity of tadpoles was decreased by temperature and salinity, which may be related to a decrease in anaerobic metabolism both in terms of sensitivity and total energy turnover through enzymatic activity. We discuss the ecological consequences, including the potential impacts on predator escape behavior promoted by changes in metabolism and locomotor performance in an early stage of development of this species.

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.

Geographical variation in genetic structure of an Atlantic Coastal Forest frog reveals regional differences in habitat stability.

Climatic oscillations throughout the Pleistocene combined with geological and topographic complexity resulted in extreme habitat heterogeneity along the Atlantic coast of Brazil. Inferring how these historic landscape patterns have structured the current diversity of the region's biota is important both for our understanding of the factors promoting diversification, as well as the conservation of this biodiversity hotspot. Here we evaluate potential historical scenarios of diversification in the Atlantic Coastal Forest of Brazil by investigating the population genetic structure of a frog endemic to the region. Using mitochondrial and nuclear sequences, we generated a Bayesian population-level phylogeny of the Thoropa miliaris species complex. We found deep genetic divergences among five geographically distinct clades. Southern clades were monophyletic and nested within paraphyletic northern clades. Analyses of historical demographic patterns suggest an overall north to south population expansion, likely associated with regional differences in habitat stability during the Pliocene and early Pleistocene. However, genetic structure among southern populations is less pronounced and likely represents more recent vicariant events resulting from Holocenic sea-level oscillations. Our analyses corroborate that the Atlantic Coastal Forest has been a biogeographically dynamic landscape and suggest that the high diversity of its fauna and flora resulted from a combination of climatic and geologic events from the Pliocene to the present.