Tadpole Survival Research Articles

Effects of temperature on metamorphosis and endochondral ossification in Rana chensinensis tadpoles

Temperature is one of the important factors affecting the growth, development, and metamorphosis of amphibians. Endochondral ossification during metamorphosis plays a crucial role in amphibian survival and adaptation on land. In this study, we explored the effects of different temperature treatments on the growth, development, and endochondral ossification of Rana chensinensis tadpoles during metamorphosis. The results showed that high temperature exposure may affect the skeletal development of tadpoles during metamorphosis, such as reduction of bone length and ossification of limbs, thyroid gland damage and change of ossification-related genes expression levels，and ultimately affect the movement and survival of tadpoles in the terrestrial environment. These results provide an experimental reference for further research on the effects of temperature on amphibian growth and development and provide an important theoretical basis for the decline of the amphibian population caused by temperature.

Responses to saltwater exposure vary across species, populations and life stages in anuran amphibians.

To predict the impacts of environmental change on species, we must first understand the factors that limit the present-day ranges of species. Most anuran amphibians cannot survive at elevated salinities, which may drive their distribution in coastal locations. Previous research showed that coastal Hyla cinerea are locally adapted to brackish habitats in North Carolina, USA. Although Hyla squirella and Hyla chrysoscelis both inhabit coastal wetlands nearby, they have not been observed in saline habitats. We take advantage of naturally occurring microgeographic variation in coastal wetland occupancy exhibited by these congeneric tree frog species to explore how salt exposure affects oviposition site choice, hatching success, early tadpole survival, plasma osmolality and tadpole body condition across coastal and inland locations. We observed higher survival among coastal H. cinerea tadpoles than inland H. cinerea, which corroborates previous findings. But contrary to expectations, coastal H. cinerea had lower survival than H. squirella and H. chrysoscelis, indicating that all three species may be able to persist in saline wetlands. We also observed differences in tadpole plasma osmolality across species, locations and salinities, but these differences were not associated with survival rates in salt water. Instead, coastal occupancy may be affected by stage-specific processes like higher probability of total clutch loss as shown by inland H. chrysoscelis or maladaptive egg deposition patterns as shown by inland H. squirella. Although we expected salt water to be the primary filter driving species distributions along a coastal salinity gradient, it is likely that the factors dictating anuran ranges along the coast involve stage-, species- and location-specific processes that are mediated by ecological processes and life history traits.

Effects of temperature on intestinal microbiota and lipid metabolism in Rana chensinensis tadpoles.

Climate change such as global warming is considered a major threat to amphibians. The guts of amphibians are home to trillions of microbes, which are key regulators of gastrointestinal digestion and play a crucial role in lipid metabolites. The aim of this study was to evaluate the effect of temperature change on intestinal microbiota and lipid metabolism in Rana chensinensis tadpoles. Morphological and intestinal microbiota data of R. chensinensis larvae exposed to different temperatures (15°C, 21°C, and 26°C) were measured. The results show that the warm temperature causes histological damage to the intestinal epithelium. In addition, temperature treatments alter the diversity and composition of gut microbes in R. chensinensis tadpoles. At the phylum level of intestinal microbial community, Campilobacterota was detected only in the warm group. At the genera level, unclassified_f__Enterobacteriaceae was markedly declined in the warm group but was notably enriched in the cold group. For lipid metabolism-related genes, the expression levels of GPR109A, HDAC1, and APOA-I decreased significantly in both warm and cold treatment groups, while the expression levels of CLPS and LIPASE increased significantly. Collectively, these observations demonstrated that warm and cold temperatures may reduce the immune capacity of tadpoles by changing the composition of intestinal microorganisms and the expression of genes related to lipid metabolism, affecting the survival of tadpoles.

Evidence for the Predator Attraction Hypothesis in an amphibian predator-prey system.

Many species possess damage-released chemical alarm cues that function in alerting nearby individuals to a predator attack. One hypothesis for the evolution and/or maintenance of such cues is the Predator Attraction Hypothesis, where predators, rather than prey, are the "intended" recipients of these cues. If a predator attack attracts additional predators, these secondary predators might interfere with the predation event, providing the prey with a better chance to escape. In this study, we conducted two experiments to explore this hypothesis in an amphibian predator/prey system. In Experiment 1, we found that tiger salamanders (Ambystoma mavortium) showed a foraging attraction to chemical cues from wood frog (Lithobates sylvaticus) tadpoles. Salamanders that were experienced with tadpole prey, in particular, were strongly attracted to tadpole alarm cues. In Experiment 2, we observed experimental encounters between a tadpole and either one or two salamanders. The presence of the second predator caused salamanders to increase attack speed at the cost of decreased attack accuracy (i.e., increasing the probability that the tadpole would escape attacks). We also found that the mere presence of visual and chemical cues from a second predator did not affect this speed/accuracy trade-off but did cause enough of a distraction to increase tadpole survival. Thus, our findings are consistent with the Predator Attraction Hypothesis for the evolution and/or maintenance of alarm cues.

Effects of water level variations on survival, morphological phenotype and responsiveness of tadpoles to phytohemagglutinin

To investigate the responses of morphological and physiological characteristics in tadpoles of Pelophylax nigromaculatus to various water levels, a total of 480 tadpoles with similar sizes were treated with rapid, moderate and slow desiccation. Both the metamorphosis duration and average metamorphosis survival rate of specific developmental stages were counted. Their body and internal organ sizes, as well as the responsiveness to phytohemagglutinin (PHA-P) of froglets were measured after complete metamorphosis. The results showed that the metamorphosis duration of tadpoles in the control group was the longest. The average metamorphosis survival rate of different treatment groups ranged from 72.5% to 90.8%, which was the highest in the control group and the lowest in the rapid desiccation group. Body weight (BW) and body length (BL) of froglets in control and slow desiccation groups were significantly higher than those of moderate or rapid desiccation groups. Body width, ratio of BW to BL, carcass wet weight and the wet weight index of lung and fat body were the lowest in rapid desiccation group. There was no significant difference in the wet weight index of heart, spleen, kidneys and digestive organs, as well as the length index of different segments of whole digestive tract between groups. The maximal response value of froglets to PHA-P appeared at 3 h after injection in different treatments, which was significantly higher in moderate and slow desiccation groups than the control group. There was no significant difference between rapid desiccation group and any other groups. The tadpoles of P. nigromaculatus might accelerate the metamorphosis developmental process to cope with desiccation stress, accompanying with decreased atrophic body size and weakened cell-mediated immunity, which would harm tadpoles to successfully land.

Does atrazine induce changes in predator recognition, growth, morphology, and metamorphic traits of larval skipper frogs (Euphlyctis cyanophlyctis)?

Atrazine, an info disruptor, interferes with the olfaction of aquatic organisms by impairing the chemosensory system. Consequently, it affects behavior, physiology, and growth increases mortality and infections, and suppresses the immune system of aquatic animals. In this study, we wanted to determine the sensitivity of larval Euphlyctis cyanophlyctis to different concentrations of atrazine by assessing their antipredator behavior, growth, morphology, and metamorphic traits. The results indicate that exposure to atrazine did not affect the survival of tadpoles. However, it caused retarded growth at higher concentrations. Interestingly, the antipredator behavior of tadpoles toward conspecific alarm cues decreased in a dose-dependent manner with an increase in the concentration of atrazine. Tadpoles exposed to low concentrations of atrazine had deeper, wider bodies and tails while those exposed to higher concentrations had shallower and narrower bodies with shallower tail muscles. However, at low and moderate concentrations atrazine did not affect size at metamorphosis, it extended the larval duration at higher concentrations.

Ontogeny of risk assessment and escape-hatching performance by red-eyed treefrog embryos in two threat contexts.

Arboreal embryos of red-eyed treefrogs, Agalychnis callidryas, hatch prematurely in response to hypoxia when flooded and to mechanosensory cues in snake attacks, but hatching later improves tadpole survival. We studied ontogenetic changes in risk assessment and hatching performance of embryos in response to flooding and physical disturbance. We hypothesized that risk assessment decreases as hatchling survival improves and hatching performance increases as embryos develop. Because snakes eat faster than embryos asphyxiate, we hypothesized that embryos decide to hatch sooner and hatch faster in response to mechanosensory cues. We video-recorded individual embryos hatching in response to each cue type, then compared the incidence and timing of a series of events and behaviors from cue onset to complete hatching across ages and stimuli. Latency from cue to hatching decreased developmentally in both contexts and was shorter with mechanosensory cues, but the elements contributing to those changes differed. Hypoxia assessment involved position changes, which decreased developmentally along with assessment time. Mechanosensory cue assessment occurred more rapidly, without movement, and decreased with age. The first stages of hatching, membrane rupture and head emergence, were surprisingly age independent but faster with mechanosensory cues, congruent with greater effort under more immediate risk. In contrast, body emergence and compression showed ontogenetic improvement consistent with morphological constraints but no cue effect. Both appropriate timing and effective performance of hatching are necessary for continued development. Different stages of the process vary with development and environmental context, suggesting combinations of adaptive context- and stage-dependent behavior, cue-related constraints on information acquisition, and ontogenetic constraints on elements of performance.

Effects of two firefighting chemical formulations, Phos–Chek LC95W and BlazeTamer380, on striped marsh frog (Limodynastes peronii) tadpole survival, growth, development and behaviour

Global wildfire events are projected to become more frequent and severe due to the continual threat of climate change, resulting in increasing demand for effective fire mitigation methods. Firefighting chemicals (FFCs), including retardants, foams and water enhancers, are often used to prevent the spread of wildfires. However, the impact of FFCs on wildlife and ecosystems is poorly understood. We investigated the effects of two common FFC formulations, Phos-Chek LC95W and BlazeTamer380, on tadpole survival, growth, development and swimming behaviour. Tadpoles of the striped marsh frog (Limnodynastes peronii) were exposed to two concentrations of either Phos-Chek (0.25 and 1 g/L) or BlazeTamer (0.05 and 0.2 g/L) for 16 days. The highest concentration of Phos–Chek was lethal to tadpoles, with mortalities gradually increasing over time and only 8% of animals surviving to day 16. Both FFCs influenced the growth and development of tadpoles, though effects were more severe in tadpoles exposed to the Phos-Chek formulation. Phos-Chek was found to completely stop tadpole growth and development over the 16-day exposure, whereas BlazeTamer significantly delayed growth and development in comparison to controls. Nevertheless, treatments had no apparent effect on tadpole movement patterns and swimming activity. Greater toxicity caused by the Phos-Chek treatment likely relates to the increased ammonia and altered water quality parameters. Runoff or accidental application of commonly used FFCs into small waterways may therefore have important ramifications for aquatic biota.

Effects of isolated and combined exposures of Boana curupi (Anura: Hylidae) tadpoles to environmental doses of trichlorfon and ultraviolet radiation

The biodiversity collapse strongly affects the amphibian group and many factors have been pointed out as catalytic agents. It is estimated that several events in the amphibian population decline worldwide may have been caused by the interaction of multiple drivers. Thus, this study aimed to evaluate the stressful effects of the exposure to environmental doses of trichlorfon (TCF) pesticide (0.5 μg/L; and an additional 100-fold concentration of 50 µg/L) and ultraviolet radiation (UV) (184.0 kJ/m² of UVA and 3.4 kJ/m² of UVB, which correspond to 5% of the daily dose) in tadpoles of the Boana curupi species (Anura: Hylidae). The isolated and combined exposures to TCF happened within 24 h of acute treatments under laboratory-controlled conditions. In the combined treatments, we adopted three different moments (M) of tadpole irradiation from the beginning of the exposures to TCF (0 h – M1; 12 h – M2; and 24 h – M3). Then, we evaluated tadpole survival, change in morphological characters, induction of apoptotic cells, lipid peroxidation (LPO), protein carbonyl content (PCC), glutathione S-transferase (GST), non-protein thiols (NPSH), and acetylcholinesterase (AChE), as well as the induction of genomic DNA (gDNA) damage. UVB treatment alone resulted in high mortality, along with a high level of apoptosis induction. Both UVA, UVB, and TCF increased LPO, PC, and AChE, while decreased GST activity. Regarding co-exposures, the most striking effect was observed in the interaction between UVB and TCF, which surprisingly decreased UVB-induced tadpole mortality, apoptosis, and gDNA damage. These results reinforce the B. curupi sensitivity to solar UVB radiation and indicate a complex response in face of UVB interaction with TCF, which may be related to activation of DNA repair pathways and/or inhibition of apoptosis, decreasing UVB-induced tadpole mortality.

Phenotypic variation in Xenopus laevis tadpoles from contrasting climatic regimes is the result of adaptation and plasticity.

Phenotypic variations between populations often correlate with climatic variables. Determining the presence of phenotypic plasticity and local adaptation of a species to different environments over a large spatial scale can provide insight on the persistence of a species across its range. Amphibians, and in particular their larvae, are good models for studies of phenotypic variation as they are especially sensitive to their immediate environment. Few studies have attempted to determine the mechanisms that drive phenotypic variation between populations of a single amphibian species over a large spatial scale especially across contrasting climatic regimes. The African clawed frog, Xenopus laevis, occurs in two regions with contrasting rainfall regimes in southern Africa. We hypothesised that the phenotypic variation of life-history traits of X. laevis tadpoles emerges from a combination of plastic and genetic responses. We predictedthat plasticity wouldallow the development of tadpoles from both regions in each environment. We also predicted thatlocal adaptation of larval traits would drive the differentiation of reaction norms between populations and lower survival in tadpoles reared away from their home environment. We measured growth, time to metamorphosis, and survival in a reciprocal transplant experiment using outdoor mesocosms. Supporting our prediction, we found that the measured variation of all traits was explained by both adaptation and plasticity. However, the reaction norms differed between populations suggesting adaptive and asymmetric plasticity. All tadpoles experienced lower survival when translocated, but only translocated tadpoles from the winter rainfall region matched survival of local tadpoles. This has implications for the dynamics of translocated X. laevis into novel environments, especially from the winter rainfall region. Our discovery of their asymmetric capacity to overcome novel environmental conditions by phenotypic plasticity alone provides insight into their invasion success.

Variable temperature regimes and wetland salinity reduce performance of juvenile wood frogs

On a changing planet, amphibians must respond to weather events shifting in frequency and magnitude, and to how those temperature and precipitation changes interact with other anthropogenic disturbances that modify amphibian habitat. To understand how drastic changes in environmental conditions affect wood frog tadpoles, we tested five temperature manipulations, including Ambient (water temperatures tracking daily air temperatures), Elevated (+ 3 °C above ambient), Nightly (removal of nightly lows), Spike (+ 6 °C above ambient every third week), and Flux (alternating ambient and + 3 °C weekly) crossed with Low Salt (specific conductivity: 109-207 µS-cm) and High Salt (1900-2000 µS-cm). We replicated each of the ten resulting treatments four times. High-salinity conditions produced larger metamorphs than low-salinity conditions. Tadpole survival was reduced only by the Spike treatment (P = 0.017). Elevated temperatures did not shorten larval periods; time to metamorphosis did not differ among temperature treatments (P = 0.328). We retained 135 recently metamorphosed frogs in outdoor terrestrial enclosures for 10months to investigate larval environment carryover effects. Juvenile frogs grew larger in low-density terrestrial enclosures than high density (P = 0.015) and frogs from Ambient LowSalt larval conditions grew and survived better than frogs from manipulated larval conditions. Frogs from HighSalt larval conditions had lower survival than frogs from LowSalt conditions. Our results suggest that anthropogenic disturbances to larval environmental conditions can affect both larval and post-metamorphic individuals, with detrimental carryover effects of high-salinity larval conditions not emerging until the juvenile life stage.

Strigea robusta (Digenea: Strigeidae) infection effects on the gonadal structure and limb malformation in toad early development.

The anomaly P is a mass morphological anomaly reported in some water frog populations across Europe. It was found that polydactyly is only a mild attenuated form of heavy cases of the anomaly P syndrome, which have strong deformations of the hindlimbs and, partly, forelimbs. It was shown that the anomaly P is caused by the trematode Strigea robusta and this syndrome can be considered as a special case of strigeosis in amphibians. The anomaly P for a long time considered to be specific for water frogs of the genus Pelophylax. Herein, we describe polydactyly and heavy forms of the anomaly P syndrome in toads of the genera Bufo and Bufotes, as a result of exposure to S.robusta cercariae. A total of 150 tadpoles of Bufo bufo, 60 tadpoles of Bufotes viridis, and 60 tadpoles of Bufotesbaturae were divided into fiveexperimental and fourcontrol groups (30 tadpoles in each group). All anomalies in the toads were similar to those observed in water frogs. The survival of tadpoles in the experimental groups was 76%. The anomaly P was observed in 57.9% of toad tadpoles (51.8% of mild forms and 6.1% of heavy forms). The occurrence of the anomaly P varied among groups from 19% to 78%. Heavy forms of the anomaly P were found in all experimental groups. We described rare asymmetrical cases of the anomaly P. According to severe modification of limb morphology, we supposed changes of gonadal morphology (any modifications of the germ and somatic cells). The gonadal development of infected tadpoles was however the same as in uninfected toad tadpoles, and heterochromatin distribution within gonocytes had no differences as well. It seems like the parasite doesn't have any effect on the gonadal development of the toads. The lack of heavy forms in natural populations of toads, as well as a development of gonads were discussed.

Stage-Specific Environmental Correlates of Reproductive Success in Boreal Toads (Anaxyrus boreas boreas)

Compensatory recruitment can facilitate the persistence of populations experiencing high adult mortality. Because early life-stages of many taxa, including amphibians, are difficult to mark and recapture, sources of variation in survival at these stages often are unknown, which creates barriers to improving in situ recruitment rates. We leveraged count data and open N-mixture models to examine the environmental factors associated with the hatching of egg clutches, tadpole survival, and probability of metamorphosis in Boreal Toads (Anaxyrus boreas boreas) that inhabit pastures leased for cattle grazing in western Wyoming, USA. We conducted weekly surveys and measured a suite of environmental variables at 20 breeding ponds during May–September 2018. The hatching of egg clutches was most strongly related to pond surface area, as clutches often desiccated at smaller ponds. Weekly tadpole survival was lowest in ponds with high abundance of aquatic predators. Predation did not preclude metamorphosis, which was more strongly associated with higher dissolved oxygen and vegetation cover. Cattle grazing reduced vegetation cover in and around breeding ponds, which resulted in lower levels of dissolved oxygen. Grazing-induced habitat changes are therefore likely to negatively influence tadpole metamorphosis both via indirect effects on dissolved oxygen, and direct effects on vegetation cover, which also serves as feeding sites and escape cover from predators. We demonstrate the success of three critical phases in early life-stage development (egg hatching, tadpole survival, metamorphosis) was associated with different environmental factors. The inclusion of stage-specific responses in demographic analyses is therefore critical for a thorough understanding of what limits populations.

Small monsters: Insect predation limits reproduction of yellow‐bellied toad Bombina variegata to ponds in their earliest successional stage

Abstract The disappearance and deterioration of amphibian breeding habitats is a major cause of their global extinction. In Europe, this problem affects the yellow‐bellied toad Bombina variegata, which inhabits small ponds in the early stages of succession, where no predatory invertebrate species normally occur. Nonetheless, as habitats, small temporary ponds are highly endangered, a situation that is intensifying with urban spread. A system of 13 pairs of artificial ponds was constructed in 2012 for the breeding of B. variegata with the aim of reconnecting disjunct populations. During the 2 years of the study, one of the twin ponds was regularly cleared of emergent vegetation and invertebrates. The survival rates of the tadpoles released in the cleared and uncleared ponds were analysed. The survival rate of the tadpoles released into the cleared ponds was higher than of those introduced into the uncleared ones. Tadpole survival rates were also higher in new, single ponds, constructed in 2013. The lower survival rates of the introduced tadpoles are best explained by the presence of predatory aquatic invertebrates, particularly the large diving beetle Dytiscus marginalis. Experimentally obtained consumption rates indicate that one Dytiscus larva is capable of destroying a typical clutch of B. variegata tadpoles within a few hours, much faster than any of the other insect predators tested. The impact of freshwater invertebrates on tadpoles is frequently studied but rarely quantified. For conservation measures to be effective, quantitative standards need to be applied that describe the impact of predation in a predictable manner. Construction of artificial small ponds is broadly accepted as an important conservation measure compensating for the disappearance of natural amphibian breeding sites. This function of artificial ponds may be substantially enhanced by the periodic removal of invertebrate predators.

Ecotoxicity assessment of triphenyl phosphate (TPhP) exposure in Hoplobatrachus rugulosus tadpoles

Triphenyl phosphate (TPhP), a widely used aromatic organophosphate flame retardant, is known to accumulate in organisms through water, air, and soil, consequently, causing toxicity. This study is the first to evaluate the acute and sub-chronic toxicities of TPhP to amphibians. In the acute toxicity analysis, the 96-h median lethal concentration (LC50) for GS35 Hoplobatrachus rugulosus tadpoles was 2.893 mg/L, and the 10% effect concentration (EC10) was 289 μg/L. After two weeks of exposure to low TPhP concentrations, the survival and metamorphosis rates of H. rugulosus tadpoles decreased, and the metamorphosis time was prolonged as the TPhP concentration increased. The threshold concentration that affected tadpole survival and metamorphosis time was 50 μg/L and 100 μg/L, respectively. No significant differences were observed in the condition factor and hepatic somatic index of the tadpole after metamorphosis; however, tadpole body mass and TPhP concentration were negatively correlated. Further, TPhP inhibited the expressions of Cu–Zn sod and cat, thereby reducing the activities of superoxide dismutase and catalase in the tadpole liver. The threshold for affecting gene expression and enzymatic activity was 100 μg/L. These findings provide significant insights on the stress ecology of aquatic organisms.

Natural history traits of the terrestrial breeding frog Batrachyla taeniata (Anura: Batrachylidae) in wet meadows of Patagonia

ABSTRACT Anurans have diverse reproductive modes, and some species of the austral forests of Patagonia have become specialized in terrestrial reproduction. Batrachyla taeniata is one of these species: it inhabits wetlands, lays its eggs during late summer and fall in terrestrial conditions, and has overwintering tadpoles. We studied a population of B. taeniata that breeds in wet meadows surrounded by native Patagonian forest and described its life cycle parameters (breeding phenology, egg development, and coexistence with other anurans) over two different years with contrasting hydroperiods. We also analyzed the effect of predatory insects on tadpole survival. Embryo development occurred in wet soil, which may be exposed to the sun or not, and took from 48 to 63 days to hatch. Free-swimming larvae overwintered in the wetland and coexisted in spring with pond-breeding anurans that varied in composition and abundance according to the wetland hydroperiod. The survivorship of B. taeniata tadpoles was significantly greater than that of P. thaul tadpoles when both were exposed to a free belostomatid predator. We consider that the terrestrial reproductive mode and life cycle of B. taeniata enable this species to survive in the variable biotic conditions determined by hydroperiod length.

Tadpole-transporting frogs use stagnant water odor to find pools in the rainforest.

ABSTRACTBreeding sites are often a limited and ephemeral resource for rainforest frogs. This resource limitation has driven the evolution of diverse reproductive strategies that increase offspring survival. For example, poison frogs shuttle their tadpoles from terrestrial clutches to aquatic rearing sites, using various cues to assess pool suitability. Yet, how frogs find new pools is unknown. We tested the role of odor cues in the process of finding tadpole deposition sites by the poison frog Allobates femoralis. We created 60 artificial pools grouped into three conditions: stagnant water, tadpole water and clean water control. Fifteen pools were discovered within 6 days, with more tadpoles and more frogs directly observed at pools with stagnant odor cues. Our findings suggest that frogs use odor cues associated with stagnant water for the initial discovery of new breeding pools. These cues may be good indicators of pool stability and increased likelihood of tadpole survival.

Polymethylmethacrylate nanoplastics can cause developmental malformations in early life stages of Xenopus laevis

Polymethylmethacrylate (PMMA) production has increased almost 20% over the last years. With its release into the aquatic environment, its breakdown or degradation to nano dimensions (nanoplastics-NPLs) due to biological and physical/mechanical action is, theoretically, anticipated. The occurrence of PMMA-NPLs in aquatic ecosystems may thus cause adverse effects particularly to early life stages of amphibians, which may be in contact with PMMA-NPLs suspended in the water column or deposited in upper layers of the sediments. Accordingly, this work aimed at assessing the effects of PMMA-NPLs to aquatic early life stages of the model anuran species Xenopus laevis. To attain this objective, two types of toxicity assays were carried out by exposing embryos [Nieuwkoop and Faber (NF) stage 8-11] or tadpoles (NF 45) to three concentrations of PMMA-NPLs (1, 100 and 1000 μg/L): i) 96-h embryo teratogenicity assay, where survival, malformation, and total body length (BL) of embryos were assessed; and ii) 48-h feeding rate assay, where survival, feeding (FR), malformations and growth rates (body weight-BW and BL) of tadpoles were evaluated. PMMA-NPLs exposure had no significant effects on mortality, malformations of X. laevis embryos but BL was lower at 1000 μg PMMA-NPLs/L. In tadpoles, no effects on survival or FR were observed after exposure to PMMA-NPLs, but significant changes occured in BW and BL. Moreover, anatomical changes in the abdominal region (externalization of the gut) were observed in 62.5% of the tadpoles exposed to 1000 μg PMMA-NPLs/L. Despite the lack of knowledge regarding the environmental levels of NPLs, it is expected that sediments constitute a sink for these contaminants, where they can become available for organisms that, like tadpoles, feed on the organic matter at the surface of sediments. Considering the continuous release and subsequent accumulation of PMMA, the malformations obtained in the feeding assays suggest that, in the future, these nano-polymers may constitute a risk for aquatic life stages of amphibians.

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.

Evaluation of the genotoxic, mutagenic, and histopathological hepatic effects of polyoxyethylene amine (POEA) and glyphosate on Dendropsophus minutus tadpoles

Herbicides improve the productivity of a monoculture by eliminating weeds, although they may also be toxic and have negative effects on non-target organisms, such as amphibians. The present study evaluated the genotoxic, mutagenic, and histopathological hepatic responses of Dendropsophus minutus tadpoles to acute exposure (96 h) to the herbicide glyphosate (GLY, 65, 130, 260 and 520 μg/L) and the surfactant polyoxyethylene amine (POEA, 1.25, 2.5, 5 and 10 μg/L). On average, 174 % more genomic damage was observed in the tadpoles exposed to all concentrations of POEA in comparison with the control, while up to seven times more micronuclei were recorded, on average, at a concentration of 5 μg/L of POEA. All the individuals exposed to 10 μg/L of POEA died. The tadpoles exposed to GLY presented 165 % more DNA damage than the control, on average, at the highest concentrations (260 and 520 μg/L), and up to six times more micronuclei at 520 μg/L. The Erythrocyte Nuclear Abnormality test (ENA) detected a relatively high frequency of cells with lobed nuclei in the tadpoles expose to POEA at 5 μg/L and binucleated cells in those exposed to GLY at 520 μg/L. The hepatic histopathological observations revealed several types of lesions in the tadpoles exposed to both GLY and POEA. Overall, then, the results of the study indicate that both GLY and POEA have potential genotoxic, mutagenic, and hepatotoxic effects in D. minutus tadpoles. We emphasize the need for further studies to monitor the amphibian populations, such as those of D. minutus, which breed in aquatic environments associated with agricultural areas. The release of pollutants into natural habitats may have significant long-term impacts on the survival of anuran tadpoles.