Natural modulation of redox status throughout the ontogeny of Amazon frog Physalaemus ephippifer (Anura, Leptodactylidae).

During their development, amphibians undergo various physiological processes that may affect their susceptibility to environmental pollutants. Naturally occurring fluctuations caused by developmental events are often overlooked in ecotoxicological studies. Our aim is to investigate how biomarkers of oxidative stress are modulated at different stages of larval development in the Amazonian amphibian species, Physalaemus ephippifer. The premetamorphosis, prometamorphosis and metamorphic climax stages were used to analyze total antioxidant capacity (ACAP), glutathione S-transferase (GST) activity, lipid peroxidation (LPO) levels and the expression of genes nrf2, gst, gsr (glutathione reductase) and gclc (glycine-cysteine ligase, catalytic subunit). Although there was no difference in ACAP and the genes expression among the studied stages, individuals from the premetamorphosis and prometamorphosis showed higher GST activity than ones under the climax. LPO levels were highest in individuals from the metamorphic climax. The present study suggests that the oxidative status changes during ontogeny of P. ephippifer tadpoles, especially during the metamorphic climax, the most demanding developmental phase. Variations in the redox balance at different developmental stages may lead to a divergent response to pollution. Therefore, we recommend that studies using anuran larvae as biomonitors consider possible physiological differences during ontogeny in their respective analyses.

Patterns of allometric and heterochronic changes in the early ontogeny of species of Physalaemus (Anura: Leptodactylidae).

We studied the relationship between shape, size, and developmental time in the embryonic ontogeny of 15 species of the frog genus Physalaemus. As in other anuran exotrophic embryos, shape changes are correlated with size increase and mainly concern tail elongation, decrease in body height, and increase in fin height. Size ranges and developmental times vary interspecifically. Embryos of the P. signifer Clade and the P. gracilis Group are among the largest, are slightly peramorphic, and develop fast regarding congeneric species. Embryos of P. cicada combine the smallest sizes with fast development and the most peramorphic shapes. The paedomorphic shapes of embryos of P. biligonigerus and P. henselii groups are correlated with fast vs. slow developmental times respectively. Trajectories in the P. cuvieri Group are diverse and in general differ in size and developmental time. The embryos of P. cristinae and from the Argentinean lineage of P. cuvieri stand out with the longest development. Sequences of developmental events are overall conserved in the genus, and main differences concern mouthpart ontogeny. This study constitutes the first attempt to evaluate morphological, allometric, and heterochronic parameters of the early ontogeny of anurans and how these can vary and contribute to diversification in taxonomic groups.

Anurans from the upper basins of the Rio Grande and Rio Preto: eastern portion of the Serra da Mantiqueira Environmental Protection Area, state of Minas Gerais, Brazil / Anfíbios anuros das bacias hidrográficas do alto rio Grande e alto rio Preto: porção leste da Área de Proteção Ambiental da Serra da Mantiqueira, estado de Minas Gerais, Brasil

The Neotropical zone stands out for having the greatest richness of amphibians in the world. About 625 species of anurans are known on Atlantic Forest. The southeastern mountainous regions of Brazil called Serra do Mar and Serra da Mantiqueira, are very important areas for the diversity of anurans in the country because they shelter remaining strata of the Atlantic Forest that have several endemic species. The present study aims to expand the knowledge about the Serra da Mantiqueira anurans, specifically from the Rio Preto and Rio Grande upper basins. The study area comprised 10 sample centers. We realized 38 campaigns, lasting two days each. During this period, we used active search as visual and auditory cues. The passive search was comprised of pitfall traps. Bioacoustic analyses were used to determine the cryptic species. The sufficiency of the sampling effort was estimated through the accumulation curve using Bootstrap and Jackknife-1 as estimators. During the study 55 species were found, 40 of them in the basin of the Rio Preto, and 44 in the basin of the Rio Grande. Twenty-nine species were found in both areas. The Hylidae family was predominant, followed by Leptodactylidae. In addition to species of the Atlantic Forest, the place houses typical components of the Cerrado biome and altitudinal swamps with unexpected sympatries. This inventory expands the area of occurrence of Scinax duartei, a species considered threatened. Although adjacent to the oldest national park in Brazil, the composition of anuran species of this area was unknown until then.

A região Neotropical se destaca por possuir a maior riqueza de anfíbios do mundo. Cerca de 625 espécies de anuros são conhecidas na Mata Atlântica. As regiões montanhosas do sudeste do Brasil, denominadas Serra do Mar e Serra da Mantiqueira, são áreas muito importantes para a diversidade de anuros no país, pois abrigam estratos remanescentes da Mata Atlântica que possuem diversas espécies endêmicas. O presente estudo visa ampliar o conhecimento sobre os anuros da Serra da Mantiqueira, especificamente das altas bacias dos rios Preto e Grande. A área de estudo compreendeu 10 centros amostrais. Realizamos 38 campanhas, com duração de dois dias cada. Nesse período, utilizamos a busca ativa como pistas visuais e auditivas. A busca passiva foi composta por armadilhas de queda. Análises bioacústicas foram usadas para determinar as espécies crípticas. A suficiência do esforço amostral foi estimada através da curva de acumulação usando Bootstrap e Jackknife-1 como estimadores. Durante o estudo foram encontradas 55 espécies, sendo 40 delas na bacia do Rio Preto e 44 na bacia do Rio Grande. Vinte e nove espécies foram encontradas em ambas as bacias. A família Hylidae foi predominante, seguida por Leptodactylidae. Além de espécies da Mata Atlântica, o local abriga componentes típicos do bioma Cerrado e brejos de altitude com simpatrias inesperadas. Este inventário amplia a área de ocorrência de Scinax duartei, espécie considerada ameaçada. Embora adjacente ao parque nacional mais antigo do Brasil, a composição de espécies de anuros dessa área era desconhecida até então.

The tadpole of an insular population of Cycloramphus boraceiensis Heyer, 1983 (Anura: Cycloramphidae) with a review of larval descriptions for species in the genus

A literature survey for the external morphology of tadpoles for species in the genus Cycloramphus Tschudi, 1838, which is mainly represented by larval descriptions, resulted in finding some inconsistent character descriptions. In a few cases, some of the information is either not presented or mistakenly reported; in some cases, the illustrations provided the information not present in the descriptions. Here in we use a sample of tadpoles of an insular population identified as Cycloramphus boraceiensis, present a description for it, compare it to the original larval description for this species, and using it as a model, present a comparative review of the other larval descriptions for Cycloramphus tadpoles. We evaluate that most of the mistakes we found in the literature are associated to the unique morphology of these semiterrestrial larvae and some of their seemingly adaptation to their microhabitats, which for the majority of the known species is a film of water running associated to or near streams.(AU)

Geographic variation in shape and size of anuran tadpoles: Interpopulation comparisons in Scinax fuscovarius (Anura, Hylidae).

Among anuran species with biphasic life cycle, the occurrence of intraspecific larval morphotypes has been related to variations in developmental time, diet, geographical variation, or response to predators. Here, we evaluated the external morphological variation of larvae among three populations, located more than 270 km apart, of the anuran hylid Scinax fuscovarius by linear and geometric methods, to elucidate the presence of geographically different morphs. Comparisons targeted development, growth, and external morphology. Studied populations exhibited differences in reproductive seasonality, growth rate, timing of development, shape, and size. Shape and size comparisons revealed two well-differentiated morphs, one of them shared by the two closest populations. Morphological differences evidenced a smaller and depressed form of the entire body plan in the most distant population, which showed continuous reproduction throughout the rainy season and under more unpredictable conditions. We interpret the occurrence of the two different larval morphs in S. fuscovarius as a by-product of local geographical conditions, and discuss on possible associations with biotic and abiotic factors cues.

Synchrotron microtomography applied to the volumetric analysis of internal structures of Thoropa miliaris tadpoles.

Amphibians are models for studying applied ecological issues such as habitat loss, pollution, disease, and global climate change due to their sensitivity and vulnerability to changes in the environment. Developmental series of amphibians are informative about their biology, and X-ray based 3D reconstruction holds promise for quantifying morphological changes during growth-some with a direct impact on the possibility of an experimental investigation on several of the ecological topics listed above. However, 3D resolution and discrimination of their soft tissues have been difficult with traditional X-ray computed tomography, without time-consuming contrast staining. Tomographic data were initially performed (pre-processing and reconstruction) using the open-source software tool SYRMEP Tomo Project. Data processing and analysis of the reconstructed tomography volumes were conducted using the segmentation semi-automatic settings of the software Avizo Fire 8, which provide information about each investigated tissues, organs or bone elements. Hence, volumetric analyses were carried out to quantify the development of structures in different tadpole developmental stages. Our work shows that synchrotron X-ray microtomography using phase-contrast mode resolves the edges of the internal tissues (as well as overall tadpole morphology), facilitating the segmentation of the investigated tissues. Reconstruction algorithms and segmentation software played an important role in the qualitative and quantitative analysis of each target structure of the Thoropa miliaris tadpole at different stages of development, providing information on volume, shape and length. The use of the synchrotron X-ray microtomography setup of the SYRMEP beamline of Elettra Synchrotron, in phase-contrast mode, allows access to volumetric data for bone formation, eye development, nervous system and notochordal changes during the development (ontogeny) of tadpoles of a cycloramphid frog Thoropa miliaris. As key elements in the normal development of these and any other frog tadpole, the application of such a comparative ontogenetic study, may hold interest to researchers in experimental and environmental disciplines.

Variation in the growth and development of the hind limbs in frogs of the genus Telmatobius (Anura: Telmatobiidae).

There are remarkable interspecific differences in the sizes of the larvae of Andean frogs of the genus Telmatobius. This size variation seems to be associated with the duration of the larval stage and may affect the hind-limb morphology in Telmatobius. Larval, juvenile, and adult Telmatobius rubigo and T. oxycephalus were examined to determine the variation in relative sizes of hind-limb elements, their growth patterns during postmetamorphic life, and skeletal ontogeny. The results showed that the proportionately shorter hind limbs of T. rubigo relative to those of T. oxycephalus are associated with the protracted development and ossification of hind limbs during the prolonged larval life of T. rubigo. Postmetamorphically, the hind limbs grew faster than the body in juveniles of both species in contrast to the relative growth rates of the hind limbs and bodies of the adults. The growth phase of juvenile T. rubigo seems shorter than that of juvenile T. oxycephalus; possibly, this heightens the difference in the relative lengths of hind limbs after metamorphosis. Temperature affects the effects of thyroid hormone on growth and development, and T. rubigo lives at much higher, colder elevations than does T. oxycephalus. It is not clear whether the developmental differences described here are plastic (i.e., environmentally induced) or genetically fixed in each species.

Herbicides employed in sugarcane plantations have lethal and sublethal effects to larval Boana pardalis (Amphibia, Hylidae).

The increasing demand for biofuels favored the expansion of sugarcane and, as a consequence, in the consumption of pesticides in Brazil. Amphibians are subject to pesticide exposure for occurring in or around sugarcane fields, and for breeding at the onset of the rainy season when pesticide consumption is common. We tested the hypothesis that herbicides used in sugarcane crops, although employed for weed control and manipulated at doses recommended by the manufacturers, can cause lethal and sublethal effects on amphibian larvae. Boana pardalis was exposed to glyphosate, ametryn, 2,4-D, metribuzin and acetochlor which account to up to 2/3 of the volume of herbicides employed in sugarcane production. High mortality was observed following prolonged exposure to ametryn (76%), acetochlor (68%) and glyphosate (15%); ametryn in addition significantly reduced activity rates and slowed developmental and growth rates. AChE activity was surprisingly stimulated by glyphosate, ametryn and 2,4-D, and GST activity by ametryn and acetochlor. Some of these sublethal effects, including the decrease in activity, growth and developmental rates, may have important consequences for individual performance for extending the larval period, and hence the risk of dessication, in the temporary and semi-permanent ponds where the species develops. Future studies should seek additional realism towards a risk analysis of the environmental contamination by herbicides through experiments manipulating not only active ingredients but also commercial formulations, as well as interactions among contaminants and other environmental stressors across the entire life cycle of native amphibian species.

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.

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.

Histology and microscopic anatomy of the thyroid gland during the larval development of Pseudis platensis (Anura, Hylidae).

Several hormones regulate anuran larval development, most notably thyroid hormones (THs). In anurans, metamorphosis fails when the thyroid gland is absent or inactivated, resulting in giant tadpoles. Larval gigantism occurs naturally in neotropical frogs of the genus Pseudis as a result of a prolonged larval period. Its thyroid function is poorly investigated and the focus of this study. We describe qualitative and quantitative variations in larval development for field-captured specimens of Pseudis platensis and compare those to the development of two sympatric species, Phyllomedusa sauvagii and Pithecopus azureus, which have small tadpoles and a shorter larval period. We describe morphological changes in the thyroid glands of larval and adult specimens. In contrast to other species with similar ecological requirements, P. platensis exhibits distinct glandular activity. During premetamorphosis, there was little or no thyroid activity, a period in which the tadpole reached 70% of its maximum size. Development and degree of activity of the thyroid gland determine the duration of the early stages of the larval period. Thyroid gland histology in tadpoles appears to correlate with the TH activity, and in turn with the diversity in anuran life history transitions.

First record of growth rings for 11 native subtropical anuran species of South America.

Skeletochronology is the most accurate method to estimate a population age structure. The methodology is based on the analysis of secondary bone tissue in order to count growth rings. This study aimed to provide initial data, allowing researchers to further work out in the age of individuals and populations, sampling evidence of the presence of growth rings in 11 native species (representing nine families) of a subtropical region of southern Brazil. Four bone samples of each specimen were used to perform the skeletochronological analysis: the penultimate phalanges of the 3rd and 4th fingers, the humerus, and the femur. The presence of growth rings was confirmed in the periosteal layer of the bones of all analyzed species. In comparison with phalanges, growth rings of humeri and femora are more irregular and less distinguishable. This is the first record of growth rings to the native species herein analised. The skeletochronology was proved to be an effective tool in determining the age of anuran amphibians from a subtropical region, since this environment presents well defined climatic seasonality.

Mechanisms of Convergent Egg Provisioning in Poison Frogs.

Parental provisioning of offspring with physiological products (nursing) occurs in many animals, yet little is known about the neuroendocrine basis of nursing in non-mammalian species. Within amphibians, maternal provisioning has evolved multiple times, with mothers of some species feeding unfertilized eggs to their developing offspring until tadpoles complete metamorphosis [1-3]. We conducted field studies in Ecuador and Madagascar to ask whether convergence at the behavioral level provides similar benefits to offspring and relies on shared neural mechanisms in dendrobatid and mantellid poison frogs. At an ecological level, we found that nursing allows poison frogs to provide chemical defenses to their tadpoles in both species. At the neural level, nursing was associated with increased activity in the lateral septum and preoptic area, demonstrating recruitment of shared brain regions in the convergent evolution of nursing within frogs and across vertebrates [4]. In contrast, only mantellids showed increased oxytocin neuron activity akin to that in nursing mammals [5], suggesting evolutionary versatility in molecular mechanisms. Our findings demonstrate that maternal provisioning provides similar potential benefits to offspring and relies on similar brain regions in poison frog species with convergently evolved toxicity and maternal care. VIDEO ABSTRACT.

Effect of soil disturbance by agricultural activities on the life history traits of monkey frog (Pithecopus azureus).

We assessed whether soil disturbance by agricultural activity influences the growth, development, and survival of individuals in the larval, metamorphic, and postmetamorphic stages of amphibians. Tadpoles of Pithecopus azureus (Cope, 1862) were reared in microcosms assembled with soil from two sites, a pristine site and a rice field. For 5 weeks, we recorded tadpole growth and development as well as physicochemical variable of the water: temperature, conductivity, dissolved oxygen, and pH. The results show that rice field soil produced a level of acidification in the water that influenced the growth and development rates of tadpoles. Tadpoles reared in rice soil had a significantly lower growth rate and body length, and during a specified period, the development rate of the tadpole was significantly lower than that of tadpoles in pristine soil. Overall, tadpoles in rice soil took 3 days longer to reach metamorphosis and 1 additional day to complete metamorphosis compared with tadpoles exposed to pristine soil. Our study shows that disturbed soils modify the physicochemical conditions of temporary ponds, impacting on the initial life stage of the anurans.

Postmetamorphic ontogenetic allometry and the evolution of skull shape in Nest-building frogs Leptodactylus (Anura: Leptodactylidae).

Allometry constitutes an important source of morphological variation. However, its influence in head development in anurans has been poorly explored. By using geometric morphometrics followed by statistical and comparative methods we analyzed patterns of allometric change during cranial postmetamorphic ontogeny in species of Nest-building frogs Leptodactylus (Leptodactylidae). We found that the anuran skull is not a static structure, and allometry plays an important role in defining its shape in this group. Similar to other groups with biphasic life-cycle, and following a general trend in vertebrates, ontogenetic changes mostly involve rearrangement in rostral, otoccipital, and suspensorium regions. Ontogenetic transformations are paralleled by shape changes associated with evolutionary change in size, such that the skulls of species of different intrageneric groups are scaled to each other, and small and large species show patterns of paedomorphic/peramorphic features, respectively. Allometric trajectories producing those phenotypes are highly evolvable though, with shape change direction and magnitude varying widely among clades, and irrespective of changes in absolute body size. These results reinforce the importance of large-scale comparisons of growth patterns to understand the plasticity, evolution, and polarity of morphological changes in different clades.

Lethal toxicity of the herbicides acetochlor, ametryn, glyphosate and metribuzin to tropical frog larvae.

Despite the high amphibian biodiversity and increasing pesticide use in tropical countries, knowledge on the sensitivity of tropical amphibians to pesticides remains limited. The aim of this study was to evaluate the acute toxicity of the active ingredients of four of the main herbicides used in Brazilian sugarcane production to tadpoles of two tropical frog species: Physalaemus cuvieri and Hypsiboas pardalis. The calculated 96 h-LC50 (median lethal concentration; in mg a.s./L) values for P. cuvieri and H. pardalis were 4.4 and 7.8 (acetochlor); 15 and <10 (ametryn); 115 and 106 (glyphosate); and 85 and 68 (metribuzin), respectively. These toxicity values demonstrated little interspecies variation and the toxicity of the herbicides appeared to be at least partly related with the respective octanol-water coefficient. Published acute toxicity data of fish and amphibians for herbicides were also compiled from the US-EPA ECOTOX database. These data indicated little difference in herbicide sensitivity between tropical amphibians and both non-tropical amphibians and fish. These findings indicate that temperate (fish and amphibian) herbicide toxicity data are also protective for tropical amphibians. Constraints in such extrapolations and indications for future research are discussed.

The poverty of adult morphology: Bioacoustics, genetics, and internal tadpole morphology reveal a new species of glassfrog (Anura: Centrolenidae: Ikakogi) from the Sierra Nevada de Santa Marta, Colombia.

Ikakogi is a behaviorally and morphologically intriguing genus of glassfrog. Using tadpole morphology, vocalizations, and DNA, a new species is described from the Sierra Nevada de Santa Marta (SNSM), an isolated mountain range in northern Colombia. The new taxon is the second known species of the genus Ikakogi and is morphologically identical to I. tayrona (except for some larval characters) but differs by its genetic distance (14.8% in mitochondrial encoded cytochrome b MT-CYB; ca. 371 bp) and by the dominant frequency of its advertisement call (2928-3273 Hz in contrast to 2650-2870 Hz in I. tayrona). They also differ in the number of lateral buccal floor papillae, and the position of the buccal roof arena papillae. Additionally, the new species is differentiated from all other species of Centrolenidae by the following traits: tympanum visible, vomerine teeth absent, humeral spines present in adult males, bones in life white with pale green in epiphyses, minute punctuations present on green skin dorsum, and flanks with lateral row of small, enameled dots that extend from below eye to just posterior to arm insertion. We describe the external and internal larval morphology of the new species and we redescribe the larval morphology of Ikakogi tayrona on the basis of field collected specimens representing several stages of development from early to late metamorphosis. We discuss the relevance of larval morphology for the taxonomy and systematics of Ikakogi and other centrolenid genera. Finally, we document intraspecific larval variation in meristic characters and ontogenetic changes in eye size, coloration, and labial tooth-rows formulas, and compare tadpoles of related species. Ikakogi tayrona has been proposed as the sister taxon of all other Centrolenidae; our observations and new species description offers insights about the ancestral character-states of adults, egg clutches, and larval features in this lineage of frogs.

Intraspecific developmental variation in the life cycle of the Andean Treefrog (Boana riojana): A temporal analysis.

Intraspecific variation during the anuran larval period has been analyzed mainly in relation to the timing of metamorphosis and body size at metamorphosis. However, other traits may vary as well. We examined two developmental series of Boana riojana from the same population in two consecutive years and describe intraspecific variation in larvae of this species. We discuss how variation, if present, may influence its life cycle. We found that both larval series differed in the larval period length, one twice as long as the other. This variation primarily depended on when breeding occurred, metamorphosis was achieved during late spring in both generations and at similar sizes, and only the rate of larval development during premetamorphosis varied extensively between years. This is consistent with thyroid gland activity because when it became active the developmental trajectory became more canalized. No variation of staging sequence occurred in relation to the different durations of the larval period. However, in the long-lasting series we found two different morphs. Also, integument, thyroid gland, skeleton, and testis differentiation events occurred at the same developing stages. In contrast, ovarian differentiation proceeded at the same absolute age in both series. Sexual dimorphism becomes evident within the year after metamorphosis. The intraspecific heterochrony that we describe for the larval development of B. riojana does not lead to phenotypic variation at the end of metamorphosis. We discuss the importance of analyzing growth and development independently. Each proceeds differently in time, but with an interdependence at some point, because size and shape do not vary at the end of metamorphosis.

Effect of hydric stress on locomotion and morphology of tadpoles from temporary ponds.

Many frog species reproduce in temporary ponds maintained exclusively by rainfall, thereby being exposed to drought and possibly mortality of eggs and tadpoles. Some tadpoles, however, can survive for up to 5 days out of water but few data are available regarding the effects of dehydration on their development. The aim of this study was to evaluate whether hydric stress affects the locomotor capacity and the morphology in tadpoles of two leptodactylid frog species showing different reproductive modes (Leptodactylus fuscus and Physalaemus nattereri), examining specifically: (a) difference in survival rate and body mass between tadpoles at different hydration levels, (b) the hydric stress effect on locomotor performance, (c) difference in external morphology, and (d) visceral volume among tadpoles suffering hydric stress. Tadpoles for both species were divided into two groups, one staying in 100 ml of water and the other maintained on absorbent paper with 4 ml of water for 12, 24, and 72 hr (n = 20 each). Significant differences in weight loss were found between the groups of both species, the treatment losing more weight in all stress levels. Almost half of P. nattereri tadpoles died within 36 hr of hydric stress. We found no difference in locomotor performance between groups of L. fuscus tadpoles, but significant differences in locomotor performance, tail morphometry, and visceral volume between groups of P. nattereri tadpoles. Our results suggest that hydric stress has a significant effect on locomotion and morphology of P. nattereri tadpoles but not in L. fuscus.

Monitoring the morphological integrity of neotropical anurans.

Amphibians are considered to be excellent bioindicators to their morphophysiological characteristics and life cycle. In this context, the present study investigated the morphological integrity of anuran larvae collected in preserved environments in the Emas National Park, in the municipality of Mineiros (Goiás state, Brazil), and in environments representative of the agricultural matrix of the Rio Verde region, also in Goiás, where there is a long history of the use of agricultural pesticides. Samples of water from temporary ponds, permanent dams, and veredas were analyzed for the presence of pesticides and, especially atrazine (5350 µg/L), found at significantly higher concentrations in the agricultural matrix. We observed a high percentage (approximately 10%) of morphological malformations including alterations of the fins in Boana albopunctatus and Scinax fuscovarius; alteration in oral structures in B. albopunctatus, Dematonotus muelleri, Physalaemus centralis, Physalaemus cuvieri, and Leptodactylus fuscus mainly in the tadpoles collected in the agricultural environment in comparison with those from the protected area (3.5%; P < 0.0001, χ2 31.75). However, changes in the eyes, mouth, intestines, and nostrils, as well as amelia were observed only in the agricultural environment. The vast majority of the observed malformations were associated with the tail and oral disc, which suggests that these anatomical parameters may be used as sensitive morphological biomarkers. Given these findings, we reinforce that areas of agricultural land may have a deleterious effect on the morphological integrity of the tadpoles and consequently, on their development, and that these features may be used as indicators of environmental quality and health.