Effects of temperature on growth, development, and survival of amphibian larvae: macroecological and evolutionary patterns.

Temperature affects the rate of biochemical and physiological processes in amphibians, influencing metamorphic traits. Temperature patterns, as those observed in latitudinal and altitudinal clines, may impose different challenges on amphibians depending on how species are geographically distributed. Moreover, species' response to environmental temperatures may also be phylogenetically constrained. Here, we explore the effects of acclimation to higher temperatures on tadpole survival, development, and growth, using a meta-analytical approach. We also evaluate whether the latitude and climatic variables at each collection site can explain differences in species' response to increasing temperature and whether these responses are phylogenetically conserved. Our results show that species that develop at relatively higher temperatures reach metamorphosis faster. Furthermore, absolute latitude at each collection site may partially explain heterogeneity in larval growth rate. Phylogenetic signal of traits in response to temperature indicates a non-random process in which related species resemble each other less than expected under Brownian motion evolution (BM) in all traits, except survival. The integration of studies in a meta-analytic framework allowed us to explore macroecological and macroevolutionary patterns and provided a better understanding of the effects of climate change on amphibians.

Following the footsteps of Burmeister's leaf frog (Phyllomedusa burmeisteri) in the Atlantic forest of Brazil.

Amphibians are organisms which mainly have a biphasic life cycle. When at the larval stage, their habitat is generally aquatic, and when adults, they become terrestrial. Pond-breeding amphibians are sensitive to some disturbances in their environment which lead to the decline of a population. The interactions between the species and their environment are performed through movement. Movement ecology combines and explains the movement data of organisms with biotic and abiotic factors and because of this, knowing the movement of these creatures is of great ecological importance. We used the spool-and-line methodology in individuals of the treefrog Phyllomedusa burmeisteri, at the Reserva Ecologica Michelin, located in the southern region of Bahia in Brazil to study their movement patterns in different environments inside and outside of the forest. We monitored 19 P. burmeisteri individuals that presented a mean total movement distance of 2160.76 cm (S.D. 1152.42). We found no significant difference in the trajectories of individuals in forested and open areas, as well as a positive relationship between the distances individuals moved and their weight only in forested areas. no relationship between distance and weight of these individuals. We observed that individuals followed non-linear paths and the number of steps from one place to another varied among individuals. Our movement ecology data allows us to answer questions about short-term movement patterns of Phyllomedusa burmeisteri. This is the first step to start understanding the spatial cognition of treefrogs from the Atlantic Forest and to fill gaps about life habits of these frogs. Information on the movement patterns of a species, as well as its home range can help to create conservation strategies, regarding the creation or delimitation of protected areas, for example.

Captive breeding, embryonic and larval development of Ranitomeyavariabilis (Zimmermann & Zimmermann, 1988), (Anura, Dendrobatidae).

A solid basis to address the conservation challenges of amphibians requires an increased knowledge on their natural history and biology. Recent data on reproductive modes in amphibians suggest that they are much more complex and variable than previously thought but understudied. However, detailed information on the reproductive history is especially important to fill the current knowledge gaps. Following recent taxonomic changes in Ranitomeyavariabilis, information about captive-breeding management, image-based measurements of total length and surface area of the silhouette for individuals from embryonic to metamorphic development, and detailed larval staging for captive-bred specimens are provided from French Guiana. The development of R.variabilis from the stage eight (Gosner 1960) through metamorphosis took 79 to 91 days (n = 6) with a survival rate of 46%. The developmental stages largely matched those of the generalized staging system of Gosner (1960), with differences in the stages when labia and teeth differentiation and atrophy of the oral apparatus occurred. Compared with other studies the total length of R.variabilis tadpoles was greater at given stages than those of R.variabilis from a Peruvian population and those of the sister species, R.amazonica. Other studies concerning growth curves based on surface area data revealed that R.variabilis tadpoles at peak size were larger than those of R.amazonica, R.imitator, R.reticulata, R.sirensis, and R.vanzolinii, but smaller than R.benedicta. Our results represent the first embryonic and larval staging for R.variabilis, and detailed information is provided on their initial life phases. These data may facilitate the identification of R.variabilis tadpoles in the wild, as well helping to clarify the biogeographical distribution and taxonomic arrangement of the species. In addition, knowledge is added to the captive-breeding methodology for the species.

An elusive giant: A new species of Vitreorana Guayasamin et al., 2009 (Anura: Centrolenidae) from the northern Atlantic Forest with an osteological description and comments on integumentary spicules.

We describe a new species of Vitreorana from the Atlantic Forest of southern Bahia state, in north-eastern Brazil. Vitreorana assuh sp. nov. is by far the largest species of the genus (snout-to-vent length [SVL] = 30.9 mm and 34.1 mm in one male and female, respectively; maximum SVL recorded for all other species = 28.0 mm). The new species is morphologically most similar to V. franciscana, from the Cerrado biome in Brazil. Besides its large size, the new species has, among other diagnostic characters, the presence of vomerine teeth and the occurrence of enameled iridophores in the pericardium, hepatic and urinary bladder peritonea. Furthermore, we describe and compare the complete mineralized skeleton of the new species and V. uranoscopa, and describe and discuss the variation of integumentary spicules in all species of the genus and their relevance to Vitreorana systematics.

Roundup Original DI® and thermal stress affect survival, morphology and thermal tolerance in tadpoles of Boana faber (Hylidae, Anura).

In amphibians, stressful environments can lead to accelerated metamorphosis at the expense of total length, resulting in the occurrence of morphological abnormalities. Many studies have linked the occurrence of these phenomena to the pollution of habitats by pesticides and thermal stress. Here, we assessed how exposure to Roundup Original DI® and higher constant temperatures affect the survival of Boana faber tadpoles and estimate the CL5096hs for the population. In addition, we evaluated how exposure to Roundup affects larval growth, morphology and thermal tolerance. Our findings suggest that even at sublethal doses, Roundup Original DI® may affect the survival of Boana faber larvae. There also appears to be an additive effect between Roundup and temperature increase on larval survival, however, we need to further explore this point to determine a pattern, proving to be a promising issue to be investigated in the future. We observed effects of chronic exposure to the herbicide formulation on the morphology and growth of the tadpoles, resulting in a reduction in total length and differences in the shape of the larvae. Although we did not recover any direct effects of herbicide exposure on CTMax, we did observe an upward trend in CTMax for tadpoles exposed to Roundup. Understanding how anthropogenic changes affect anuran persistence is fundamental for the management and conservation of the species and can be considered an initial step toward the formulation of legislations that regulate the use of herbicides.

DNA barcode reveals candidate species of Scinax and Ololygon (Anura: Hylidae) in Atlantic Forest.

Molecular species delimitation methods are efficient tools to identify species, including the discovery of new taxa and cryptic organisms, thus being useful to biodiversity studies. In the present work, 16S mitochondrial sequences and cytochrome oxidase I (COI) were used to evaluate the richness of species in the genus Scinax and Ololygon from a biodiversity hotspot in Atlantic Forest. A total of 109 specimens formally belonging to eight species of Scinax and three species of Ololygon were collected in 13 localities along the state of Bahia (northeastern Brazil) and one site in Espírito Santo (southeastern Brazil). Of the Scinax species collected in this study, three were morphologically differentiated from other described species and identified as putative new species (Scinax sp.1, Scinax sp.2 and Scinax sp.3). The species delimitations were inferred using three different methods: ABGD, PTP and mPTP which allowed recognizing 11 Scinax species and five Ololygon species. Scinax sp. 1, Scinax sp. 2 and Scinax sp. 3, have been confirmed as new putative species and Ololygon argyreornata possibly contains cryptic species. We suggest additional studies, including morphological and bioacoustic data to validate these new putative species.

Natural history predicts patterns of thermal vulnerability in amphibians from the Atlantic Rainforest of Brazil.

In the Brazilian Atlantic Rainforest (AF), amphibians (625 species) face habitat degradation leading to stressful thermal conditions that constrain animal activity (e.g., foraging and reproduction). Data on thermal ecology for these species are still scarce. We tested the hypothesis that environmental occupation affects the thermal tolerance of amphibian species more than their phylogenetic relationships. We evaluated patterns of thermal tolerance of 47 amphibian species by assessing critical thermal maxima and warming tolerances, relating these variables with ecological covariates (e.g., adult macro- and microhabitat and site of larval development). We used mean and maximum environmental temperature, ecological covariates, and morphological measurements in the phylogenetic generalized least squares model selection to evaluate which traits better predict thermal tolerance. We did not recover phylogenetic signal under a Brownian model; our results point to a strong association between critical thermal maxima and habitat and development site. Forest species were less tolerant to warm temperatures than open area or generalist species. Species with larvae that develop in lentic environment were more tolerant than those in lotic ones. Thus, species inhabiting forest microclimates are more vulnerable to the synergistic effect of habitat loss and climate change. We use radar charts as a quick evaluation tool for thermal risk diagnoses using aspects of natural history as axes.

Initial Phylotranscriptomic Confirmation of Homoplastic Evolution of the Conspicuous Coloration and Bufoniform Morphology of Pumpkin-Toadlets in the Genus Brachycephalus.

The genus Brachycephalus is a fascinating group of miniaturized anurans from the Brazilian Atlantic Forest, comprising the conspicuous, brightly colored pumpkin-toadlets and the cryptic flea-toads. Pumpkin-toadlets are known to contain tetrodotoxins and therefore, their bright colors may perform an aposematic function. Previous studies based on a limited number of mitochondrial and nuclear-encoded markers supported the existence of two clades containing species of pumpkin-toadlet phenotype, but deep nodes remained largely unresolved or conflicting between data sets. We use new RNAseq data of 17 individuals from nine Brachycephalus species to infer their evolutionary relationships from a phylogenomic perspective. Analyses of almost 5300 nuclear-encoded ortholog protein-coding genes and full mitochondrial genomes confirmed the existence of two separate pumpkin-toadlet clades, suggesting the convergent evolution (or multiple reversals) of the bufoniform morphology, conspicuous coloration, and probably toxicity. In addition, the study of the mitochondrial gene order revealed that three species (B. hermogenesi, B. pitanga, and B. rotenbergae) display translocations of different tRNAs (NCY and CYA) from the WANCY tRNA cluster to a position between the genes ATP6 and COIII, showing a new mitochondrial gene order arrangement for vertebrates. The newly clarified phylogeny suggests that Brachycephalus has the potential to become a promising model taxon to understand the evolution of coloration, body plan and toxicity. Given that toxicity information is available for only few species of Brachycephalus, without data for any flea-toad species, we also emphasize the need for a wider screening of toxicity across species, together with more in-depth functional and ecological study of their phenotypes.

The phylogeny of Dendropsophini (Anura: Hylidae: Hylinae).

The relationships of the hyline tribe Dendropsophini remain poorly studied, with most published analyses dealing with few of the species groups of Dendropsophus. In order to test the monophyly of Dendropsophini, its genera, and the species groups currently recognized in Dendropsophus, we performed a total evidence phylogenetic analysis. The molecular dataset included sequences of three mitochondrial and five nuclear genes from 210 terminals, including 12 outgroup species, the two species of Xenohyla, and 93 of the 108 recognized species of Dendropsophus. The phenomic dataset includes 46 terminals, one per species (34 Dendropsophus, one Xenohyla, and 11 outgroup species). Our results corroborate the monophyly of Dendropsophini and the reciprocal monophyly of Dendropsophus and Xenohyla. Some species groups of Dendropsophus are paraphyletic (the D. microcephalus, D. minimus, and D. parviceps groups, and the D. rubicundulus clade). On the basis of our results, we recognize nine species groups; for three of them (D. leucophyllatus, D. microcephalus, and D. parviceps groups) we recognize some nominal clades to highlight specific morphology or relationships and facilitate species taxonomy. We further discuss the evolution of oviposition site selection, where our results show multiple instances of independent evolution of terrestrial egg clutches during the evolutionary history of Dendropsophus.

The phylogeny of the Casque-headed Treefrogs (Hylidae: Hylinae: Lophyohylini).

The South American and West Indian Casque-headed Treefrogs (Hylidae: Hylinae: Lophyohylini) include 85 species. These are notably diverse in morphology (e.g. disparate levels of cranial hyperossification) and life history (e.g. different reproductive modes, chemical defences), have a wide distribution, and occupy habitats from the tropical rainforests to semiarid scrubland. In this paper, we present a phylogenetic analysis of this hylid tribe based on sequence fragments of up to five mitochondrial (12S, 16S, ND1, COI, Cytb) and six nuclear genes (POMC, RAG-1, RHOD, SIAH, TNS3, TYR). We included most of its species (> 96%), in addition to a number of new species. Our results indicate: (i) the paraphyly of Trachycephalus with respect to Aparasphenodon venezolanus; (ii) the nonmonophyly of Aparasphenodon, with Argenteohyla siemersi, Corythomantis galeata and Nyctimantis rugiceps nested within it, and Ap. venezolanus nested within Trachycephalus; (iii) the polyphyly of Corythomantis; (iv) the nonmonophyly of the recognized species groups of Phyllodytes; and (v) a pervasive low support for the deep relationships among the major clades of Lophyohylini, including C. greeningi and the monotypic genera Itapotihyla and Phytotriades. To remedy the nonmonophyly of Aparasphenodon, Corythomantis, and Trachycephalus, we redefined Nyctimantis to include Aparasphenodon (with the exception of Ap. venezolanus, which we transferred to Trachycephalus), Argenteohyla, and C. galeata. Additionally, our results indicate the need for taxonomic work in the following clades: (i) Trachycephalus dibernardoi and Tr. imitatrix; (ii) Tr. atlas, Tr. mambaiensis and Tr. nigromaculatus; and (iii) Phyllodytes. On the basis of our phylogenetic results, we analyzed the evolution of skull hyperossification and reproductive biology, with emphasis on the multiple independent origins of phytotelm breeding, in the context of Anura. We also analyzed the inter-related aspects of chemical defences, venom delivery, phragmotic behaviour, co-ossification, and prevention of evaporative water loss.

A new species of Scinax Wagler (Hylidae: Scinaxini) from the tropical forests of Northeastern Brazil.

We describe a new species of the Scinax ruber clade from Northeastern Brazil that occurs in widely separated geographic areas in the Atlantic Forest of southern Bahia state and the Highland Humid Forest of Serra de Baturité, northeast Ceará state. Scinax tropicalia sp. nov. (holotype coordinates: -14.795694°, -39.172645°) is diagnosed from all 75 currently recognize species of the S. ruber clade by bioacoustical and morphological adult traits, such as duration (0.11-0.31 s) and dominant frequency (1.59-1.85 kHz) of the advertisement call, snout shape rounded, nearly rounded, or semi-circular in dorsal view and rounded to slightly protruding in profile, bilobate vocal sac, absence of pectoral glands and spicule-shaped papillary epidermal projections on nuptial pads, and color pattern on the dorsum of body and hidden surfaces of hindlimbs.

Quaternary climatic fluctuations influence the demographic history of two species of sky-island endemic amphibians in the Neotropics.

We evaluated the role of Quaternary climatic fluctuations on the demographic history and population structure of amphibian species endemic to the 'campo rupestre' in the Neotropics, evaluating their distributional shifts, demographic changes, and lineage formation from the end of Pleistocene to present. We chose two anurans endemic to the high-elevation 'campo rupestre' in the Espinhaço Range (ER) in northeastern and southeastern Brazil (Bokermannohyla alvarengai and Bokermannohyla oxente), as models to test the role of Quaternary climatic fluctuations over their distribution range in this region. We collected tissue samples throughout their distribution range and used statistical phylogeography to examine processes of divergence and population demography. We generated spatial-temporal reconstructions using Bayesian inference in a coalescent framework in combination with hind-cast projections of species distribution models (SDMs). We also used the results and literature information to test alternative diversification scenarios via approximate Bayesian computation (ABC). Our results show that Quaternary climatic fluctuations influenced the geographic ranges of both species showing population expansion during the last glacial maximum (LGM) and range contraction during interglacial periods, as inferred from selected ABC models and from past projections of SDMs. We recovered Pleistocene diversification for both species occuring in distinctly unique periods for each taxon. An older and range-restricted lineage was recovered in a geographically isolated geological massif, deserving conservation and further taxonomic study. The diversification and distribution of these amphibian species endemic to the Neotropical 'campo rupestre' were influenced by Quaternary climatic fluctuations. The expansion of cold adapted species restricted to higher elevations during glacial periods and their concomitant retraction during interglacial periods may have been crucial for producing patterns of species richness and endemism along elevation gradients in tropical and subtropical domains. Such processes may influence the evolution of the biota distributed in heterogeneous landscapes with varied topography.

Snakes of the Pernambuco Endemism Center, Brazil: diversity, natural history and conservation.

The Atlantic Forest is one of the largest and richest tropical rainforests on the planet, being one of the 25 world priorities for conservation. The Atlantic Forest portion located north of the São Francisco River corresponds to the Pernambuco Endemism Center (PEC). We describe the snake composition of the PEC, providing information about the diversity, natural history and geographical distribution of the species, based on records from five scientific collections and additional information from the literature. A total of 78 species of snakes distributed in eight families was registered in the Pernambuco Endemism Center. The Caatinga is the Brazilian biome that most shares species with the PEC, followed by Cerrado. On the other hand, seven species are considered endemic of this region. Most of the snake species in the PEC have been registered in forest (94.8%), followed by "Brejos Nordestinos" (46.1%), Tabuleiros (43.5%), Restingas (14.1%) and Mangroves (5.1%). The PEC snake fauna includes mainly terrestrial species (60.2%) and cryptozoic and/or fossorial species (21.7%), but also presents a high richness of semi-arboreal and arboreal species (29.5%). Vertebrates are the main food item consumed by the species (78% of species), among the main prey are mammals, lizards, and amphibians. Most species show a strictly nocturnal activity period (50%), followed by strictly diurnal (38%). The PEC is the most degraded and least known region of the Atlantic Forest, yet it has revealed a high richness of snake species, including seven endemic species. It is emphasized that regional conservation efforts need to be intensified, because few forests in the region are formally protected, and the majority consist of small and poorly protected fragments, which means that many species in the region may be in risk of extinction.

Amphibians and reptiles of Parque Nacional da Serra das Lontras: an important center of endemism within the Atlantic Forest in southern Bahia, Brazil.

Information gaps about species distribution hamper the evaluation of conservation status and decisions on biodiversity conservation, affecting to a greater extent, areas with high species richness and endemism. In this context, biological inventories are an important tool to fill these gaps by providing data on the composition, richness, and abundance of species in each locality. The Parque Nacional da Serra das Lontras (PNSL) protects various mountain range just up 1000 m. in altitude, and, together with other conservation units, forms an ecological corridor in the southern part of the state of Bahia, within the Atlantic Forest hotspot. We conducted systematic samplings on transects, and opportunistic records in ponds and streams, in order to record amphibian and reptile species in the PNSL. We complement the sampling with the information available in the literature and in scientific collections. A total of 100 species (49 amphibians and 51 reptiles) was recorded, 53 of them endemic to the Atlantic Forest, 13 to the state of Bahia, and two known only from the PNSL. Hylidae was the most diverse family of amphibians (22 spp.) and Colubridae of reptiles (33 spp.). New information on the distribution and natural history of these species is provided, many of which have not yet been assessed by the IUCN while others have already been categorized as at risk of extinction at the regional level. Results confirm the high species richness and rates of endemism in southern Bahia and highlight the importance of protecting high altitude areas for the preservation of evolutionary and ecological processes within the Atlantic Forest.

The advertisement call of the rare casque-headed frog Nyctimantis galeata (Anura: Hylidae) from its type locality, Morro do Chapéu, Bahia, Brazil.

Nyctimantis Boulenger, 1882 was considered a monotypic genus for over a century. In a recent phylogenetic analysis of the Lophyohylini tribe with more than 96% of its representatives, Nyctimantis was redefined with the well-supported inclusion of some species from the genus Aparasphenodon, Argenteohyla and Corythomantis (Blotto et al. 2020). Therefore, the genus Nyctimantis currently consists of seven described species: N. arapapa (Pimenta, Napoli Haddad, 2009); N. bokermanni (Pombal, 1993); N. brunoi (Miranda-Ribeiro, 1920); N. galeata (Pombal, Menezes, Fontes, Nunes, Rocha Van Sluys, 2012); N. pomba (Assis, Santana, Silva, Quintela Feio, 2013); N. rugiceps Boulenger (1882) and N. siemersi (Mertens, 1937).

A new large canopy-dwelling species of Phyllodytes Wagler, 1930 (Anura, Hylidae) from the Atlantic Forest of the state of Bahia, Northeastern Brazil.

The known diversity of treefrogs of the genus Phyllodytes has rapidly increased in recent years, currently comprising 14 species. Recent field work in the Atlantic Rainforest of the state of Bahia lead to the discovery of a new large species of Phyllodytes which is herein described based on multiple evidence including morphological, acoustical and genetic data. Phyllodytes sp. nov. is one of the largest species within the genus and presents immaculate yellowish dorsum and limbs. The advertisement call of the species is composed of 7-31 notes (half pulsed/pulsatile-half harmonic) with frequency-modulated harmonics. Phyllodytes sp. nov. has a karyotype of 2n = 22 chromosomes, as also found in other species of the genus. Genetic distance values of the 16S mitochondrial rRNA among Phyllodytes sp. nov. and its congeners range between 6.4 to 10.2%. The description of another new species for this state reinforces the need for further taxonomic work with Phyllodytes in this region that has been revealed as a priority area for research and conservation of this genus.

Chromosome evolution in Lophyohylini (Amphibia, Anura, Hylinae).

The hyline tribe Lophyohylini includes 87 species of treefrogs, of which cytogenetics aspects have been studied in less than 20% of them. In order to evaluate the evolution of some of its chromosome characters (NOR position, C-bands, and DAPI/CMA3 bands), we studied the karyotypes of 21 lophyohylines, 16 of them for the first time, and analyzed them in a phylogenetic context. Most species showed similar karyotypes regarding chromosome number (2n = 24) and morphology (FN = 48), excepting Phyllodytes edelmoi and Osteocephalus buckleyi with 2n = 22 (FN = 44) and 2n = 28 (FN = 50), respectively. The NOR location was variable among species and provided valuable phylogenetic information. This marker was located in pair 11 in all species of Trachycephalus, Itapotihyla langsdorffii, and Nyctimantis arapapa, representing the plesiomorphic condition of Lophyohylini. Besides, other apomorphic states were recovered for the clades comprising N. rugiceps and N. siemersi (NOR in pair 5), and Dryaderces pearsoni, Osteocephalus, and Osteopilus (NOR in pair 9). Phyllodytes presented variation for NORs position; they were in pair 2 in P. edelmoi, pair 7 in P. melanomystax, and pair 8 in P. gyrinaethes and P. praeceptor. Polymorphisms in size, number, and activity of this marker were observed for N. siemersi, Osteocephalus fuscifacies, and some species of Trachycephalus. Remarkably, in N. siemersi NORs were detected on a single chromosome in the two specimens studied by this technique, raising the question of how this complex polymorphism is maintained. Interstitial telomeric sequences were found in P. edelmoi, P. melanomystax, and Osteocephalus buckleyi, and their presence seems to be not related to the chromosome reorganization events. Finally, some species showed spontaneous rearrangements, possibly as a consequence of an uncommon phenomenon in anuran cytogenetics: the presence of fragile sites or secondary constrictions not associated with NORs. We propose that this rare feature would have played an important role in the evolution of this group of frogs. From the evidence obtained in this and previous studies, we conclude that Lophyohylini presents a complex chromosome evolution.

The advertisement call of Proceratophrys redacta (Anura, Odontophrynidae).

The genus Proceratophrys Miranda-Ribeiro, 1920, frogs from the family Odontophrynidae Lynch, currently contains 41 nominal species with poorly resolved phylogenetic relationships (Frost 2019; Mângia et al. 2018). Molecular data from 15 and 18 Proceratophrys species support the monophyletic hypothesis of the genus (Teixeira-Jr et al. 2012; Dias et al. 2013; respectively) but do not fully resolve the phylogenetic relationship among the species. Currently, there are advertisement call parameters provided for 31 species of Proceratophrys in the literature (Ferreira et al. 2016; Andrade et al. 2018; Mângia et al. 2018; Nascimento et al. 2019). The Proceratophrys advertisement call is composed of pulsed notes, amplitude modulation, short to medium duration calls (0.05-4 s) and only one note in almost all species (see Nascimento et al. 2019 for details). Considering the importance of bioacoustics for taxonomy (Köhler et al. 2017), further descriptions of advertisement calls for the genus could also improve our understanding of the species' relationships.

Redescription of the tadpole of Leptodactylus flavopictus Lutz, 1926 (Anura: Leptodactylidae) from Pilar do Sul, São Paulo, Brazil.

Frogs of the subfamily Leptodactylinae Werner can be found from southern Texas (USA) to southern Brazil, including the northern Antilles (Frost 2019). This subfamily is composed of 98 recognized species and the genus Leptodactylus Fitzinger contains the largest number of species (74 sp.) (Frost 2019). A study conducted by de Sá et al. (2014) provided a phylogeny of Leptodactylus based on molecular analysis, adult and larval morphology, and behavioral data. Currently, four species groups are recognized within this genus: L. fuscus group, L. pentadactylus group, L. latrans group, and L. melanonotus group (de Sá et al. 2014).