Tadpole Development Research Articles

A computational model approach to assess the effect of climate change on the growth and development of tadpoles

All of the environmental conditions in nature act on an organism simultaneously. However, in experimental studies of the factors influencing metamorphosis, each factor needs to be examined individually in order to disentangle its specific effects. However, it is challenging to then build properly integrated models which include data on all of the different factors evaluated in different experiments. This study set out to develop a predictive model which could synthesize the results of several experiments on survival, development and growth of Natterjack toad (Epidalea calamita) tadpole guilds. The proposed Population Dynamic P System (PDP) model enables estimates of growth and development during the larval phase, under different environmental conditions, weather conditions, predator density, and pond characteristics and management. The architecture of the model allows the inclusion of an indefinite number of parameters and interactions, with all inputs interacting in parallel, and enables solutions to complex modeling approaches. Using the model with a range of field data, we found that the importance of predation pressure on Natterjack toad tadpole guilds exceeds the potential effects of variations in temperature and precipitation. The impact of introduced invasive predators therefore arguably poses the greatest threat to this species. This type of model holds promise as a reliable management and conservation tool for this and other species, especially where interactions between environmental factors make the impacts of individual factors difficult to predict.

Impact of desiccation pre-exposure on deltamethrin-induced oxidative stress in Bombina variegata juveniles

Global warming represents a severe threat to existing ecosystems, especially for anuran tadpoles who encounter significant fluctuations in their habitats. Decreasing water levels in permanent and temporary water bodies is a significant risk for larval survival or fitness. On the other hand, the natural environment of amphibians is extremely polluted by various xenobiotics. This study evaluated how pre-exposure of Bombina variegata tadpoles to chronic environmental stress (desiccation) modulates the biochemical response of juvenile individuals to following acute chemical stressor (pesticide deltamethrin). Our results demonstrated that individually applied pesticide changed the thiol and lipid status of the treated juveniles but animals subjected solely to desiccation pressure were more tolerant to free radicals and showed no induction of lipid peroxidation. Comparison of juveniles exposed to deltamethrin revealed that desiccation pretreatment during the larval stage of development modified cellular protection in the juveniles. Higher activities of CAT, GSH-Px and GR were recorded in the pre-exposed group, as well as a lower degree of lipid peroxidation relative to the group that was not pre-exposed to low water stress. Pre-desiccated groups displayed the greatest range of coordination of investigated antioxidant parameters, supported by Pearson's correlations. Activation of the GSH-redox system is a significant marker in juveniles against stress caused by desiccation and a chemical stressor. The stressful environment experienced during tadpole development produced an adaptive reaction to subsequent exposure to another stressor in juveniles. To develop relevant management and conservation strategies, more studies of the interactive effects of environmental and chemical stressors are necessary.

Experimental confirmation of effects of leaf litter type and light on tadpole performance for two priority amphibians

AbstractIsolated wetlands embedded within longleaf pine savannas support a high proportion of regional biodiversity including many amphibian species. Today, remnant isolated wetlands are often overgrown and hydrologically altered due to fire exclusion or incompatible, cool season fire regimes. In the absence of warm season fires when wetlands are dry, shrubs and trees succeed herbaceous plants, which alters wetland productivity via effects on light and detritus quality. We used a factorial aquatic mesocosm study to test the effects of altered detritus and shade on the growth, development, and survival of tadpoles of two priority amphibian species: gopher frogs (Rana capito) and ornate chorus frogs (Pseudacris ornata). Gopher frog survival was higher among maidencane, sedge, and pine treatments compared to oak and sweetgum treatments. While gopher frog larval periods were lowest in the sedge treatment, there was a nominal general effect of litter type on gopher frog larval periods, growth rates, and mass at metamorphosis. Shading had a nominal and inconsistent effect on gopher frog growth rates, but did extend larval periods in all litter treatments, decreased survival in all litter treatments except oak, and decreased mass at metamorphosis in all litter treatments except pine and sweetgum. Ornate chorus frog survival was minimally affected by shading and litter treatments, but growth rates and mass at metamorphosis were highest in maidencane and sedge treatments, and larval periods were extended with shading in all litter treatments. Shading also decreased growth rates in maidencane and sedge litters and decreased mass at metamorphosis in pine and sweetgum litters. Our results demonstrate that succession of isolated wetlands can reduce tadpole performance for two priority species both through changes in leaf litter and shading, though the effect on survival, larval growth, larval period, and size at metamorphosis can differ between species. These results support management recommendations to restore and maintain open canopy, grassy conditions in isolated wetlands for conservation of priority amphibian species.

Earlier Snowmelt Advances Breeding Phenology of the Common Frog (Rana temporaria) but Increases the Risk of Frost Exposure and Wetland Drying

The alarming decline of amphibians around the world calls for complementary studies to better understand their responses to climate change. In mountain environments, water resources linked to snowmelt play a major role in allowing amphibians to complete tadpole metamorphosis. As snow cover duration has significantly decreased since the 1970s, amphibian populations could be strongly impacted by climate warming, and even more in high elevation sites where air temperatures are increasing at a higher rate than at low elevation. In this context, we investigated common frog (Rana temporaria) breeding phenology at two different elevations and explored the threats that this species faces in a climate change context. Our objectives were to understand how environmental variables influence the timing of breeding phenology of the common frog, and explore the threats that amphibians face in the context of climate change in mountain areas. To address these questions, we collected 11 years (2009–2019) of data on egg-spawning date, tadpole development stages, snowmelt date, air temperature, rainfall and drying up of wetland pools at ∼1,300 and ∼1,900 m a.s.l. in the French Alps. We found an advancement of the egg-spawning date and snowmelt date at low elevation but a delay at high elevations for both variables. Our results demonstrated a strong positive relationship between egg-spawning date and snowmelt date at both elevations. We also observed that the risk of frost exposure increased faster at high elevation as egg-spawning date advanced than at low elevation, and that drying up of wetland pools led to tadpole mortality at the high elevation site. Within the context of climate change, egg-spawning date is expected to happen earlier in the future and eggs and tadpoles of common frogs may face higher risk of frost exposure, while wetland drying may lead to higher larval mortality. However, population dynamics studies are needed to test these hypotheses and to assess impacts at the population level. Our results highlight climate-related threats to common frog populations in mountain environments, but additional research should be conducted to forecast how climate change may benefit or harm amphibian populations, and inform conservation and land management plans in the future.

Effects of Trematode Parasites on Snails and Northern Leopard Frogs (Lithobates pipiens) in Pesticide-Exposed Mesocosm Communities

Chemical contamination of aquatic environments is widespread, but we have a limited understanding of how contaminants alter critical host–parasite interactions that can influence disease dynamics. We manipulated Northern Leopard Frog (Lithobates pipiens) exposure to pesticides (no pesticides, the insecticide Bacillus thuringiensis israelensis [Bti], or the herbicide atrazine) and trematode-infected (Ribeiroia ondatrae and Echinostoma spp.) snails in outdoor mesocosms. Bti exposure extended host larval period, and atrazine exposure had a nonsignificant trend toward reducing host survival; however, neither pesticide influenced parasite success nor magnified the effects of parasites on their hosts. Parasites negatively influenced tadpole development and, by metamorphosis, parasitized frogs had severe limb deformities and greater mass than unparasitized frogs. The greater mass in parasitized frogs may have resulted from reduced competition between tadpoles and snails for algal resources because parasites decreased snail abundance in mesocosms. Reduced competition between tadpoles and snails may offset the direct negative effects of trematodes on tadpoles, enabling them to survive with high infection intensities. Trematodes may further facilitate their own success by inducing limb deformities that likely increase anuran consumption by definitive hosts. Our results demonstrate how common pesticides and parasites impact amphibians and suggest that, at environmentally relevant concentrations, these pesticides may not dramatically alter host–parasite dynamics.

Altered temperature affect body condition and endochondral ossification in Bufo gargarizans tadpoles.

Bufo gargarizans is one kind of economic animals with higher medicinal value in China. In this study, B. gargarizans (Bufo gargarizans) tadpoles were reared at three different water temperature (15, 22 and 29°C) from Gosner stages 28-46. We investigated the effects of temperature on growth, development, survival, metamorphic duration, size and skeletal ossification at Gosner stage 40, 42, and 46, as well as thyroid tissue reached metamorphic climax (Gs42). Besides, we examined the transcription levels of endochondral ossification-related genes in hind limb at metamorphic climax (Gs42). Our results showed that the growth and development of tadpoles conform to the temperature-size rule (TSR). While warm temperature resulted in the decrease in body size and hind limb length, and shorten larval period, cold temperature led to increase in body size and hind limb length but prolonged larval period. Histological examinations revealed that warm and cold temperatures caused damage to thyroid tissue. Also, warm and cold temperatures inhibited the degree of ossification with the double staining methodology. Additionally, the real-time PCR results suggested that warm and cold temperatures significantly up-regulated Runx2, VEGF and VEGFR mRNA levels, and down-regulated TRβ, MMP9, MMP13 and Runx3 mRNA levels. The up-regulation of Dio2 level and down-regulation of Dio3 level were observed in warm temperature. TRα mRNA level was significantly increased in warm temperature, but decreased in cold temperature. Collectively, these observations demonstrated that warm and cold temperatures affected endochondral ossification in B. gargarizans tadpoles, which might influence their capacity to terrestrial locomotion.

Left High and Dry: Froth Nesting Allows Eggs of the Anuran Amphibian to Complete Embryogenesis in the Absence of Free-Standing Water

For amphibians that oviposit in temporary aquatic systems, there is a high risk of desiccation-induced offspring mortality when water evaporates prior to the completion of embryo or tadpole development. Such a strong selective pressure has led to the evolution of a variety of traits in reproducing females and their offspring to improve the odds of reproductive success when free-standing water is temporarily available. Herein, we describe an adaptive function of froth nesting in the Sandpaper Frog, Lechriodus fletcheri, as a possible maternal strategy for protecting embryos from the immediate risk of desiccating in highly ephemeral pools that frequently dry prior to hatching. Field observations revealed that embryos located near the core of nests remained alive and continued to develop for several days after becoming stranded out of water due to declining water levels, with embryo viability maintained long enough for additional rainfall to recharge pools and support hatching of tadpoles into water in some cases. In laboratory trials, the proportion of embryos surviving in nests exposed to desiccating conditions was positively correlated with nest volume, while the rate of water loss relative to nest mass declined, both of which are likely a function of reduced surface area relative to volume in larger nests. We suggest that the encapsulation of embryos within an aerated mucus shields them from desiccation by trapping moisture around their external surfaces. As embryos of L. fletcheri complete development rapidly, the ability for the froth nest to protect against desiccation for several days may allow embryogenesis to be completed largely out of water despite the larval phase not being terrestrial. These results suggest froth nesting has played an important role in facilitating this species' use of ephemeral habitats that most other amphibians with aquatic reproductive modes are incapable of exploiting.

Ontogenetic changes in activity, locomotion and behavioural complexity in tadpoles

Abstract Metamorphosis is a widespread developmental process that involves considerable changes in morphology, habitat use, ecology and behaviour between early developmental (larval) stages and adult forms. Among amphibians, anuran larvae (tadpoles) undergo massive morphological and ecological changes during their development, with early stages characterized by somatic growth, whereas more conspicuous changes (i.e. metamorphosis) occur later during development. In this study, we examined how locomotor and behavioural traits covary with morphology (body size) and metamorphosis (hindlimb and forelimb development) across developmental stages in spined toad (Bufo spinosus) tadpoles. As expected, we found that locomotion and behaviour undergo significant changes during tadpole development. These changes are curvilinear across developmental stages, with a phase of increasing activity and locomotion followed by a phase of stasis and/or reduction in locomotion and behavioural complexity. All the metrics we investigated indicate that the peak of activity and associated behaviour is situated at a pivotal stage when somatic growth decreases and significant morphological changes occur (i.e. hindlimb growth). Future studies that aim to investigate determinants of locomotion should include developmental stages as covariates in order to assess whether the sensitivity of locomotion to environmental variables changes across developmental stages.

Influence of prey diet quality on predator-induced traits in wood frog tadpoles (Lithobates sylvaticus)

Abstract Diet quality and predation are two critical factors in determining the growth and development of organisms. Various anurans are susceptible to phenotypic changes influenced by these factors. Yet, few studies examined prey diet quality as potential influence over predator-induced traits. Using wood frog tadpoles (Lithobates sylvaticus) as a model species, we investigated the effects of three diet compositions (plant-based, animal-based, omnivorous) crossed with presence or absence of chemical cues from predatory dragonfly larvae (Aeshnidae). After 35 days, we recorded 11 morphological measurements, Gosner stage, and intestinal length of tadpoles to assess phenotypic changes under the six different experimental conditions. Our results showed the additive effects of both diet quality and predator chemical cue without detection of interactions between the two. Tadpoles receiving the omnivorous diet grew and developed faster with wider denticle rows than those receiving the plant or animal diets. The growth and development of tadpoles receiving only the animal diet were significantly hindered. These results emphasize the importance of diet quality in the growth and development of larval wood frogs. Chemical cues from predators significantly reduced tadpole body size but, in contrast to previous findings, did not affect tail size. Our experimental procedure of providing water containing predator and injured conspecific chemical cues on a weekly basis likely provided relatively weak predation risk perceived by tadpoles compared to previous studies using caged predators. The predator environment in our experiment, however, represents one ecologically relevant scenario in which predation risk is not urgent.

Exposure impacts of Imazapyr formulation on larval development and thyroid histology of Xenopus laevis.

There are increased concerns about the thyroidal effects of many anthropogenic substances in the environment. These substances include agricultural pesticides and industrial and pharmaceutical chemicals among others. Their potential thyroidal effects are of serious health and ecological concerns, as thyroid hormones mediate numerous physiological processes, including growth regulation, general metabolism and metamorphosis in metamorphic animals. This study assessed thyroidal activities of Arsenal formulation (Imazapyr) at environmentally relevant concentrations of 0.5, 2.0 and 3.5 mg/L following a Xenopus metamorphosis assay (XEMA). The result shows that the Arsenal formulation significantly delayed the tadpole development, reduced the hind-limb length (HLL) and increased the whole-body mass (WBM) at a concentration of 3.5 mg/L relative to the control exposure. In histopathology, the formulation increased the epithelium height, at all exposure concentrations, but reduced the colloidal area at 0.5 and 2 mg/L, respectively, and the gland area at 2 mg/L relative to the control. Consequently, the Arsenal formulation is thyroid-active at environmentally relevant concentrations and poses a threat to both human and wildlife, especially metamorphic organisms. With this exposure impact, more studies are imperative to further characterise other endocrine-disrupting potential of this formulation, while future applications should be reduced or restricted to less risk environment, if it cannot be stopped from sensitive aquatic systems.

The Effects of Ditch Management in Agroecosystems on Embryonic and Tadpole Survival, Growth, and Development of Northern Leopard Frogs (Lithobates pipiens).

Agricultural drainage ditches help remove excess water from fields and provide habitat for wildlife. Drainage ditch management, which includes various forms of vegetation clearing and sediment dredging, can variably affect the ecological function of these systems. To determine whether ditch conditions following dredging/vegetation clearing management affected the survival, growth, and development of embryos and tadpoles of northern leopard frogs (Lithobates pipiens), we conducted three field studies using in situ cages over 2years. We measured nutrients, pesticides, and other water quality properties in vegetated/unmanaged (i.e., no clearing or dredging) and newly cleared/dredged (i.e., treeless, then dredged), clay-bottomed drainage ditches in a river basin in Eastern Ontario, Canada. Nutrients, atrazine, and total neonicotinoid concentrations were generally lower at the cleared/dredged sites, whereas glyphosate was at higher concentrations. In contrast, water-quality variables measured in situ, particularly temperature, dissolved oxygen, and turbidity, tended to be higher in the cleared/dredged sites. Total phosphorous and total organic carbon concentrations at all sites were above the recommended limits for amphibian assays. No significant differences were detected in the survival, hatching success, or development of embryos among the ditch management treatments, but premature hatching was observed at one vegetated/unmanaged site where high specific conductivity may have been formative. We found the cleared/dredged sites supported earlier tadpole growth and development, likely as a result of the higher water temperatures. Increased temperature may have offset other growth/development stressors, such as those related to water chemistry. However, the long-term consequences of these differences on amphibian populations requires further study.

Cadmium-induced toxicity to amphibian tadpoles might be exacerbated by alkaline not acidic pH level

Heavy metal pollution in natural water bodies generally interacting with other environmental stressors produces toxic effects on aquatic organisms. However, toxicological studies exploring interactive effects of these stressors are still limited. Here, tadpoles of the Zhenhai brown frog (Rana zhenhaiensis) were exposed to a 3 × 3 factorial combination, with three cadmium (Cd) concentrations (0, 10 and 100 μg/L) and three pH levels (5.0, 7.23 and 9.0) throughout the developmental period to assess combined toxic effects of Cd × pH on tadpole growth, development and physiology. Nearly all measured traits [including survival, metamorphosis and abnormality rate, metamorphosis time, post-metamorphic size, hepatic metal content, locomotor performance, antioxidant enzyme activity, and erythrocytic nuclear abnormality (ENA) frequency] were affected by Cd exposure, indicating notable Cd-induced toxicity to R. zhenhaiensis tadpoles. The pH level and its interaction with Cd also had significant impacts on most measured traits, such as survival rate, metamorphosis time, froglet jumping distance, hepatic Cd content, ENA frequency. Acidic (or alkaline) environment itself was toxic to tadpoles. However, high pH (but not low pH) level appeared to exacerbate Cd-induced toxicity to tadpoles. Excess free hydrogen ions under acidic environments might inhibit Cd2+ ions binding to cell surface, which reduced Cd accumulation in tissues. Under alkaline environments, other forms of Cd complexes in the aqueous phase probably contributed to promoting Cd accumulation. Our results indicated that Cd exposure could interact with different pH levels, producing diverse combined toxicities to amphibian larvae.

Vitamin C dietary supplementation influence tadpoles of bullfrog Lithobates catesbeianus reared in low water temperature

Vitamin C supplementation is important for the growth and development of bullfrog tadpoles under optimum water temperature conditions. Therefore, an experiment was carried out to evaluate the effects of vitamin C supplementation on the diet of bullfrog tadpoles at a low temperature. A total of 480 tadpoles with a mean weight of 0.078 g were distributed in 12 aquariums each containing 40 L of water in a closed water recirculation system. The experimental design was entirely randomized with four treatments (0, 150, 300, and 600 mg kg-1 of L-ascorbic acid monophosphate) and three replicates. The productive performance was measured by the weight gain, feed conversion, diet consumption, protein efficiency, carcass yield, hepatosomatic index, viscerosomatic index, visceral fat index, dry matter, and ethereal carcass extract. The water temperature during the experimental period was 21.74 ± 0.43 °C. Vitamin C supplementation did not influence carcass yield and viscerosomatic index. However, there was a quadratic effect of vitamin C supplementation on the weight gain, apparent feed conversion, protein efficiency, visceral fat index, hepatosomatic index, and ethereal carcass extract. Based on these results, bullfrog tadpoles should be supplemented with 600 mg vitamin C kg-1 of the diet when subjected to water temperatures of around 22 °C.

Complex trade-offs in oviposition site selection in a cannibalistic frog

For oviparous animals such as amphibians, the presence or absence of conspecifics can influence site selection, with each life history stage potentially influencing the decision-making process in a different manner. In the present study, we tested the effect of conspecific life history stage on oviposition site selection in the sandpaper frog, Lechriodus fletcheri. We quantified preferences for spawning in pools that were unused by conspecifics, as well as pools with conspecific spawn or tadpoles present, or both. The study included two ecological contexts: a system of naturally occurring breeding pools and a field experiment with 40 artificial pools that controlled for habitat variation. Mothers preferred to oviposit in pools that already contained conspecific spawn, despite the likelihood of intense resource competition upon offspring hatching. This could be due to the potential benefits obtained by providing offspring access to a significant nutrient supply upon hatching via cannibalism of conspecific tadpoles and might be an important adaptation for completing tadpole development in highly ephemeral and resource-limited pools prior to desiccation. In contrast, mothers avoided ovipositing in pools with conspecific tadpoles, a probable adaptation to avoid their own offspring from becoming victims of cannibalism themselves prior to hatching. Such nuances in the effect of conspecific presence on offspring survival highlight the often complex decision-making process that amphibians need to make when selecting oviposition sites, as well as the influence cannibalism can have on the evolution of reproductive behaviour.

Bottom-up effects of fungicides on tadpoles of the European common frog (Rana temporaria).

Biodiversity is under pressure worldwide, with amphibians being particularly threatened. Stressors related to human activity, such as chemicals, are contributing to this decline. It remains, however, unclear whether chemicals exhibiting a fungicidal activity could indirectly affect tadpoles that depend on microbially conditioned leaf litter as food source. The indirect effect of fungicides (sum concentration of a fungicide mixture composed of azoxystrobin, carbendazim, cyprodinil, quinoxyfen, and tebuconazole: 100 µg/L) on tadpoles was assessed relative to leaf litter colonized by microbes in absence of fungicides (control) and a worst‐case scenario, that is leached leaf litter without microbial colonization. The quality of leaf litter as food for tadpoles of the European common frog (Rana temporaria) was characterized through neutral lipid fatty acid profiles and microbial sum parameters and verified by sublethal responses in tadpoles (i.e., feeding rate, feces production, growth, and fatty acid composition). Fungicides changed the nutritious quality of leaf litter likely through alterations in leaves’ neutral lipid fatty acid profiles (i.e., changes in some physiologically important highly unsaturated fatty acids reached more than 200%) in combination with a potential adsorption onto leaves during conditioning. These changes were reflected by differences in the development of tadpoles ultimately resulting in an earlier start of metamorphosis. Our data provide a first indication that fungicides potentially affect tadpole development indirectly through bottom‐up effects. This pathway is so far not addressed in fungicide environmental risk assessment and merits further attention.

A bird-like genome from a frog: Mechanisms of genome size reduction in the ornate burrowing frog, Platyplectrum ornatum

The diversity of genome sizes across the tree of life is of key interest in evolutionary biology. Various correlates of variation in genome size, such as accumulation of transposable elements (TEs) or rate of DNA gain and loss, are well known, but the underlying molecular mechanisms driving or constraining genome size are poorly understood. Here, we study one of the smallest genomes among frogs characterized thus far, that of the ornate burrowing frog (Platyplectrum ornatum) from Australia, and compare it to other published frog and vertebrate genomes to examine the forces driving reduction in genome size. At ∼1.06 gigabases (Gb), the P. ornatum genome is like that of birds, revealing four major mechanisms underlying TE dynamics: reduced abundance of all major classes of TEs; increased net deletion bias in TEs; drastic reduction in intron lengths; and expansion via gene duplication of the repertoire of TE-suppressing Piwi genes, accompanied by increased expression of Piwi-interacting RNA (piRNA)-based TE-silencing pathway genes in germline cells. Transcriptomes from multiple tissues in both sexes corroborate these results and provide insight into sex-differentiation pathways in Platyplectrum Genome skimming of two closely related frog species (Lechriodus fletcheri and Limnodynastes fletcheri) confirms a reduction in TEs as a major driver of genome reduction in Platyplectrum and supports a macroevolutionary scenario of small genome size in frogs driven by convergence in life history, especially rapid tadpole development and tadpole diet. The P. ornatum genome offers a model for future comparative studies on mechanisms of genome size reduction in amphibians and vertebrates generally.

Double-edged sword: Fluoxetine and ibuprofen as development jeopardizers and apoptosis' inducers in common toad, Bufo bufo, tadpoles

Nowadays the presence of pharmaceuticals in the environment is a real problem. Ending up in aquatic environments they negatively affect non-target organisms. Considering the limited studies on the negative effects of pharmaceuticals in amphibians, a better understanding of the mechanisms underlying the sub-lethal effects of drug mixtures in wildlife is an urgent call. Representing particularly vulnerable organisms currently at risk of extinction, amphibians are perfect non-target organisms to explore the consequences of pharmaceuticals during sensitive life-stages. To address this existing research gap, the effects of two drugs, the antidepressant fluoxetine and the anti-inflammatory ibuprofen, as well as their combination has been studied. Tadpoles of Bufo bufo were exposed for seven days to two environmentally realistic concentrations of fluoxetine, ibuprofen and their mixture. The development, behavior and erythron profile were then evaluated as endpoints of exposure response. Both drugs negatively affected tadpoles' growth and development by significantly delayed their time to metamorphosis and reduced body weight. Behaviors were also impaired with a significant increase of unresponsiveness to different stimuli. Mutagenic analysis of blood revealed a significant increase in the frequency of cellular and nuclear abnormalities. Given the complexity of systems and functions affected, our work confirms the toxicological potential of fluoxetine and ibuprofen in B. bufo tadpoles by emphasizing their role as tadpole development delayers and erythrocyte apoptosis-inducers. To our knowledge, this is the first study trying to elucidate the potentially toxic effects of a mixture of an antidepressant with a non-steroidal anti-inflammatory drug using bullfrog tadpole as model organism. Both drugs interacted in impairing development and fitness in tadpoles, which might affect long-term species perpetuation and population dynamic. More in-depth research is needed to elucidate the nature of interaction and molecular mechanisms of mixed pharmaceutical compounds on non-targeted organisms.

A new reproductive mode in anurans: Natural history of Bokermannohyla astartea (Anura: Hylidae) with the description of its tadpole and vocal repertoire.

Anurans have the greatest diversity of reproductive modes among tetrapod vertebrates, with at least 41 being currently recognized. We describe a new reproductive mode for anurans, as exhibited by the Paranapiacaba Treefrog, Bokermannohyla astartea, an endemic and poorly known species of the Brazilian Atlantic Forest belonging to the B. circumdata group. We also describe other aspects of its reproductive biology, that are relevant to understanding the new reproductive mode, such as courtship behavior, spawning, and tadpoles. Additionally, we redescribe its advertisement call and extend its vocal repertoire by describing three additional call types: courtship, amplectant, and presumed territorial. The new reproductive mode exhibited by B. astartea consists of: (1) deposition of aquatic eggs in leaf-tanks of terrestrial or epiphytic bromeliads located on or over the banks of temporary or permanent streams; (2) exotrophic tadpoles remain in the leaf-tanks during initial stages of development (until Gosner stage 26), after which they presumably jump or are transported to streams after heavy rains that flood their bromeliad tanks; and (3) tadpole development completes in streams. The tadpoles of B. astartea are similar to those of other species of the B. circumdata group, although with differences in the spiracle, eyes, and oral disc. The vocal repertoire of B. astartea exhibits previously unreported acoustic complexity for the genus. Bokermannohyla astartea is the only bromeligenous species known to date among the 187 known species within the tribe Cophomantini. We further discuss evolutionary hypotheses for the origin of this novel reproductive mode.

Toxicity of Cypermethrin-chlorpyrifos and Butachlor to Tadpole of Quasipaa boulengeri

The tadpole of Quasipaa boulangeri was taken as the research object in this paper. The toxic effects of cypermethrin-chlorpyrifos and butachlor on tadpole of Q.boulangeri were studied by acute toxicity experiment and subchronic toxicity experiment in order to master the toxicity of different types of pesticides on tadpole of Q.boulangeri and provide scientific basis for amphibian protection. The results of acute toxicity experiment were analyzed by the method of probability unit regression analysis. The results of the acute toxicity experiment were analyzed by means of the probability-unit regression analysis method, and the LC50 of cypermethrin-chlorpyrifos and butachlor to the 2-month-old frog tadpoles at 24h, 48h, 72h and 96h were 0.419, 0.357, 0.346L, 0.332 and 1.183, 1.108, 0.925 and 0.876(mg/L), respectively. In the subchronic toxicity experiment, we observed the accumulation effect of different concentrations of pesticides on tadpoles through the statistics of the growth and development of tadpoles exposed to the drugs for a long time. The results suggested that the body length and body weight of tadpoles increased slowly with the increase of time and concentration of pesticide in the solution during the experiment.

Effects of temperature on growth, development and the leptin signaling pathway of Bufo gargarizans.

Climate change is one of the most important causes of the decline in amphibians. Changes in temperature have an important effect on the growth and development and energy metabolism of amphibians. The aim of this study is to unravel the effects of temperature on the leptin signaling pathway of Bufo gargarizans and its molecular mechanisms. Our results showed that high temperature accelerated the development rate of tadpoles, but reduced body size and mass, while low temperature deferred the development of tadpoles, but increased size and mass. Both high temperature and low temperature exposure caused pathological damage of the liver in B. gargarizans. The results of RT-qPCR revealed that the high temperature treatment significantly upregulated the transcript levels of genes related to thyroid hormone (DIO2 (D2), Thyroid Hormone Receptor-α (TRα)) and the leptin signaling pathway (Leptin Receptor (LepR), Janus kinase 1 (JAK1), Janus kinase 2 (JAK2), Tyrosine kinase 2 (TYK2), Signal Transducer And Activator Of Transcription 3 (STAT3), Signal Transducer And Activator Of Transcription 3.1 (STAT3.1), and Signal Transducer And Activator Of Transcription 6 (STAT6)), while there was a decrease of mRNA expression of these genes (TRα, Thyroid Hormone Receptor-Beta (TRβ), LepR, JAK1, and TYK2) in the liver of tadpoles exposed to high temperature compared with the intermediate temperature treatment. Therefore, our results suggested that temperature extremes might interfere with the thyroid and leptin signaling pathways and affect the growth and development of B. gargarizans. Furthermore, tissue injury of the liver could occur due to exposure to temperature extremes. This work promotes public awareness of environmental protection and species conservation needs, also provides valuable experimental data and a theoretical basis for the protection of amphibians.