Frog hatchlings use early environmental cues to produce an anticipatory resource-use phenotype.

Developmental plasticity can occur at any life stage, but plasticity that acts early in development may give individuals a competitive edge later in life. Here, we asked if early (pre-feeding) exposure to a nutrient-rich resource impacts hatchling morphology in Mexican spadefoot toad tadpoles, Spea multiplicata. A distinctive carnivore morph can be induced when tadpoles eat live fairy shrimp. We investigated whether cues from shrimp--detected before individuals are capable of feeding--alter hatchling morphology such that individuals could potentially take advantage of this nutritious resource once they begin feeding. We found that hatchlings with early developmental exposure to shrimp were larger and had larger jaw muscles--traits that, at later stages, increase a tadpole's competitive ability for shrimp. These results suggest that early developmental stages can assess and respond to environmental cues by producing resource-use phenotypes appropriate for future conditions. Such anticipatory plasticity may be an important but understudied form of developmental plasticity.

Evolution of hyperossification expands skull diversity in frogs.

Frogs (Anura) are one of the most diverse vertebrate orders, comprising more than 7,000 species with a worldwide distribution and extensive ecological diversity. In contrast to other tetrapods, frogs have a highly derived body plan and simplified skull. In many lineages of anurans, increased mineralization has led to hyperossified skulls, but the function of this trait and its relationship with other aspects of head morphology are largely unexplored. Using three-dimensional morphological data from 158 species representing all frog families, we assessed wide-scale patterns of shape variation across all major lineages, reconstructed the evolutionary history of cranial hyperossification across the anuran phylogeny, and tested for relationships between ecology, skull shape, and hyperossification. Although many frogs share a conserved skull shape, several extreme forms have repeatedly evolved that commonly are associated with hyperossification, which has evolved independently more than 25 times. Variation in cranial shape is not explained by phylogenetic relatedness but is correlated with shifts in body size and ecology. The species with highly divergent, hyperossified skulls often have a specialized diet or a unique predator defense mechanism. Thus, the evolution of hyperossification has repeatedly facilitated the expansion of the head into multiple new shapes and functions.

Ontogeny of the anuran urostyle and the developmental context of evolutionary novelty.

Developmental novelties often underlie the evolutionary origins of key metazoan features. The anuran urostyle, which evolved nearly 200 MYA, is one such structure. It forms as the tail regresses during metamorphosis, when locomotion changes from an axial-driven mode in larvae to a limb-driven one in adult frogs. The urostyle comprises of a coccyx and a hypochord. The coccyx forms by fusion of caudal vertebrae and has evolved repeatedly across vertebrates. However, the contribution of an ossifying hypochord to the coccyx in anurans is unique among vertebrates and remains a developmental enigma. Here, we focus on the developmental changes that lead to the anuran urostyle, with an emphasis on understanding the ossifying hypochord. We find that the coccyx and hypochord have two different developmental histories: First, the development of the coccyx initiates before metamorphic climax whereas the ossifying hypochord undergoes rapid ossification and hypertrophy; second, thyroid hormone directly affects hypochord formation and appears to have a secondary effect on the coccygeal portion of the urostyle. The embryonic hypochord is known to play a significant role in the positioning of the dorsal aorta (DA), but the reason for hypochordal ossification remains obscure. Our results suggest that the ossifying hypochord plays a role in remodeling the DA in the newly forming adult body by partially occluding the DA in the tail. We propose that the ossifying hypochord-induced loss of the tail during metamorphosis has enabled the evolution of the unique anuran bauplan.

Transcriptomic bases of a polyphenism.

Polyphenism-in which multiple distinct phenotypes are produced from a single genotype owing to differing environmental conditions-is commonplace, but its molecular bases are poorly understood. Here, we examine the transcriptomic bases of a polyphenism in Mexican spadefoot toads (Spea multiplicata). Depending on their environment, their tadpoles develop into either a default "omnivore" morph or a novel "carnivore" morph. We compared patterns of gene expression among sibships that exhibited high versus low production of carnivores when reared in conditions that induce the carnivore morph versus those that do not. We found that production of the novel carnivore morph actually involved changes in fewer genes than did the maintenance of the default omnivore morph in the inducing environment. However, only body samples showed this pattern; head samples showed the opposite pattern. We also found that changes to lipid metabolism (especially cholesterol biosynthesis) and peroxisome contents and function might be crucial for establishing and maintaining differences between the morphs. Thus, our findings suggest that carnivore phenotype might have originally evolved following the breakdown of robustness mechanisms that maintain the default omnivore phenotype, and that the carnivore morph is developmentally regulated by lipid metabolism and peroxisomal form, function, and/or signaling. This study also serves as a springboard for further exploration into the nature and causes of plasticity in an emerging model system.

Genetic evidence for polyandry in the threatened green and golden bell frog.

Identifying which species exhibit polyandry may lead to further insights into evolutionary biology and social behaviour. However, confirming polyandry can be difficult. High-resolution genetics provides a useful means to gain evidence. Although the threatened Pelodryadid frog, the green and golden bell frog Litoria aurea, has been subject to numerous ecological studies, there is uncertainty surrounding its reproductive ecology. Polyandry has not been formally identified in L. aurea or any species within the Pelodryadidae family. We aimed to identify if there was genetic evidence of polyandry in a population occurring in a wetland complex on Kooragang Island, New South Wales. To accomplish this, we collected genetic samples of tadpoles within the same size cohort about 20-30 days after explosive breeding events. Genotypes of 14 females, nine males and 70 tadpoles were analysed with COLONY (1988 single nucleotide polymorphisms after filtering) to identify parentage, full-siblings and half-siblings. We found support for the hypothesis that L. aurea is polyandrous. Based on previous observations of multi-male matings and the narrow time periods that breeding occurred in, it is likely this species exhibits simultaneous polyandry. We discuss these results in regards to behavioural adaptive processes and avenues for further research.

Rise in temperature causes decreased fitness and higher extinction risks in endemic frogs at high altitude forested wetlands in northern Pakistan.

The larvae of some amphibian species grow and develop more slowly at lower temperature and attain larger body size at the end of metamorphosis, but this is not true for all amphibians. We have for the first time provided empirical data by using a combination of field and lab-based studies with a robust sample size (360 adults, 1000 tadpoles) for Hazara Torrent Frog (Allopaa hazarensis) and Murree Hills Frog (Nanorana vicina). In this study, we examined how tadpoles responded to different temperatures through metamorphosis, body size, developmental complications or deformities, fitness and survival. We found that the tadpoles of A. hazarensis and N. vicina showed the same response to elevated temperatures under laboratory conditions including faster metamorphosis, reduction in the body size, more frequent developmental complications or deformities such as edema and tail kinks, lower fitness and higher mortality at elevated temperatures (>26 °C). The comparison of thermal sensitivity between the two species showed that N. vicina was more sensitive to higher temperatures. Pakistan has been experiencing changes in climatic patterns, and the ecosystems in northern Pakistan are losing biodiversity due to increasing temperature, frequency of drought and intensity of floods. It is feared that these frogs may experience local extinction in future. Our findings contribute to filling the information gap regarding impacts of temperature increase on biodiversity of high altitude forested montane ecosystems and is an important contribution to future studies associating biodiversity and climate change.

Observations on spindly leg syndrome in a captive population of Andinobates geminisae.

Amphibian health problems of unknown cause limit the success of the growing number of captive breeding programs. Spindly leg syndrome (SLS) is one such disease, where affected individuals with underdeveloped limbs often require euthanization. We experimentally evaluated husbandry-related factors of SLS in a captive population of the critically endangered frog, Andinobates geminisae. SLS has been linked to tadpole nutrition, vitamin B deficiency, water filtration methods, and water quality, but few of these have been experimentally tested. We tested the effects of water filtration method and vitamin supplementation (2017) and the effects of tadpole husbandry protocol intensity (2018) on time to metamorphosis and the occurrence of SLS. We found that vitamin supplementation and reconstituted reverse osmosis filtration of tadpole rearing water significantly reduced SLS prevalence and that reduced tadpole husbandry delayed time to metamorphosis. A fortuitous accident in 2018 resulted in a decrease in the phosphate content of rearing water, which afforded us an additional opportunity to assess the influence of phosphate on calcium sequestration. We found that tadpoles that had more time to sequester calcium for ossification during development had decreased the prevalence of SLS. Taken together, our results suggest that the qualities of the water used to rear tadpoles plays an important role in the development of SLS. Specifically, filtration method, vitamin supplementation, and calcium availability of tadpole rearing water may play important roles. Focused experiments are still needed, but our findings provide important information for amphibian captive rearing programs affected by high SLS prevalence.

An integrative phylogenomic approach illuminates the evolutionary history of Old World tree frogs (Anura: Rhacophoridae).

Rhacophoridae are one of the most speciose and ecologically diverse families of amphibians. Resolution of their evolutionary relationships is key to understanding the accumulation of biodiversity, yet previous hypotheses based on Sanger sequencing exhibit much discordance amongst generic relationships. This conflict precludes the making of sound macroevolutionary conclusions. Herein, we conduct the first phylogenomic study using broad-scale sampling and sequences of 352 nuclear DNA loci obtained using anchored hybrid enrichment targeted sequencing. The robust time-calibrated phylogenetic hypothesis clarifies several long-disputed relationships and facilitates the testing of evolutionary hypotheses on spatiotemporal diversification and reproductive modes. The major extant lineages of Rhacophoridae appear to have radiated in mainland Asia, and the spatiotemporal process corresponds with several common accumulations of biodiversity in Asia. Analyses do not detect any case of "Out of Himalaya" in Rhacophoridae. All transitions of reproductive modes appear to have evolved in an ordered, gradual sequence associated with gaining independence of standing water for larval development. The different reproductive modes are phylogenetically conserved and the completion of their transitions appear to have occurred over a period of ~30 Ma, which does not fit a pattern of a rapid burst of diversification. Innovations in reproductive modes associate statistically with the uneven distribution of species-richness between clades, where higher diversification is linked to increased terrestrial modes of reproduction. These results strengthen the hypothesis that breeding innovations drive diversification by providing new opportunities for ecological release and dispersion.

The mechanics of air breathing in gray tree frog tadpoles, Hyla versicolor (Anura: Hylidae).

We describe air-breathing mechanics in gray tree frog tadpoles (Hyla versicolor). We found that H. versicolor tadpoles breathe by 'bubble-sucking', a breathing mode typically employed by tadpoles too small to break the water's surface tension, in which a bubble is drawn into the buccal cavity and compressed into the lungs. In most tadpoles, bubble-sucking is replaced by breach breathing (breaking the surface to access air) at larger body sizes. In contrast, H. versicolor tadpoles bubble-suck throughout the larval period, despite reaching body sizes at which breaching is possible. Hyla versicolor tadpoles exhibit two bubble-sucking behaviors: 'single bubble-sucking', previously described in other tadpole species, is characterized by a single suction event followed by a compression phase to fill the lungs; 'double bubble-sucking' is a novel, apparently derived form of bubble-sucking that adds a second suction event. Hyla versicolor tadpoles transition from single bubble-sucking to double bubble-sucking at approximately 5.7 mm snout-vent length (SVL), which corresponds to a period of rapid lung maturation when they transition from low to high vascularization (6.0 mm SVL). Functional, behavioral and morphological evidence suggests that double bubble-sucking increases the efficiency of pulmonary gas exchange by separating expired, deoxygenated air from freshly inspired air to prevent mixing. Hyla versicolor, and possibly other hylid tadpoles, may have specialized for bubble-sucking in order to take advantage of this increased efficiency. Single and double bubble-sucking represent two- and four-stroke ventilation systems, which we discuss in the context of other anamniote air-breathing mechanisms.

Embryonic Epidermal Lectins in Three Amphibian Species, Rana ornativentris, Bufo japonicus formosus, and Cynops pyrrhogaster.

Intelectins (Itlns) are secretory lectins found in several chordate species that recognize carbohydrates on the bacterial cell surface depending on Ca2 + . In newly hatched larvae of Rana ornativentris (R. orn), Bufo japonicus formosus (B. jpn), and Cynops pyrrhogaster (C. pyr), an anti-Itln monoclonal antibody (mAb) labeled a subset of epidermal cells in whole-mount immunocytochemical assays. In western blot analyses, the mAb identified protein bands at approximately 33-37 kDa in the larval extracts and concentrated larval culture media. Using RT-PCR and RACE techniques, we isolated cDNAs from newly hatched larvae that encoded proteins of 343 (R. orn), 336 (B. jpn), and 337 (C. pyr) amino acids having 70%, 71%, and 60% identities with that of the Xenopus laevis embryonic epidermal lectin (XEEL), respectively. The proteins, designated REEL, BEEL, and CEEL, showed characteristics conserved among reported Itln proteins, and their amino acid sequences following the signal peptides were identical to those of the N-terminal peptides determined on Itln proteins in the respective larval extracts. Recombinant REEL (rREEL), rBEEL, and rCEEL proteins produced by HEK-293T cells were homo-oligomers of 34-37 kDa subunit peptides, which were similar to the Itlns found in the newly hatched larvae. The rEELs showed carbohydrate-binding specificities similar to that of XEEL and agglutinated Escherichia coli and Staphylococcus aureus cells depending on Ca2 + . These results suggest that REEL, BEEL, and CEEL are Itlns produced and secreted by epidermal cells of R. orn, B. jpn, and C. pyr larvae, respectively, and that Itlns have a conserved role as pathogen recognition molecules in the larval innate immune system.

Tissues injury and pathological changes in Hyla intermedia juveniles after chronic larval exposure to tebuconazole.

Tebuconazole (TBZ), an azole pesticide, is one of the most frequently detected fungicides in surface water. Despite its harmful effects, mainly related to endocrine disturbance, the consequences of TBZ exposure in amphibians remain poorly understood. Here, we investigated the adverse and delayed effects of TBZ chronic exposure on a native anuran species, often inhabiting cultivated areas, the Italian tree frog (Hyla intermedia). To disclose the multiple mechanisms of action through which TBZ exerts its toxicity we exposed tadpoles over the whole larval period to two sublethal TBZ concentrations (5 and 50 µg/L), and we evaluated histological alterations in three target organs highly susceptible to xenobiotics: liver, kidney, and gonads. We also assessed morphometric and gravimetric parameters: snout-vent length (SVL), body mass (BM), liver somatic index (LSI), and gonad-mesonephros complex index (GMCI) and determined sex ratio, gonadal development, and differentiation. Our results show that TBZ induces irreversible effects on multiple target organs in H. intermedia, exerting its harmful effects through several pathological pathways, including a massive inflammatory response. Moreover, TBZ markedly affects sexual differentiation also by inducing the appearance of sexually undetermined individuals and a general delay of germ cell maturation. Given the paucity of data on the effects of TBZ in amphibians, our results will contribute to a better understanding of the environmental risk posed by this fungicide to the most endangered group of vertebrates.

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.

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.

Larger Genomes Linked to Slower Development and Loss of Late-Developing Traits.

Genome size varies widely among organisms and is known to affect vertebrate development, morphology, and physiology. In amphibians, genome size is hypothesized to contribute to loss of late-forming structures, although this hypothesis has mainly been discussed in salamanders. Here we estimated genome size for 22 anuran species and combined this novel data set with existing genome size data for an additional 234 anuran species to determine whether larger genome size is associated with loss of a late-forming anuran sensory structure, the tympanic middle ear. We established that genome size is negatively correlated with development rate across 90 anuran species and found that genome size evolution is correlated with evolutionary loss of the middle ear bone (columella) among 241 species (224 eared and 17 earless). We further tested whether the development of the tympanic middle ear could be constrained by large cell sizes and small body sizes during key stages of tympanic middle ear development (metamorphosis). Together, our evidence suggests that larger genomes, slower development rate, and smaller body sizes at metamorphosis may contribute to the loss of the anuran tympanic middle ear. We conclude that increases in anuran genome size, although less drastic than those in salamanders, may affect development of late-forming traits.

Plasticity-led evolution: evaluating the key prediction of frequency-dependent adaptation.

Plasticity-led evolution occurs when a change in the environment triggers a change in phenotype via phenotypic plasticity, and this pre-existing plasticity is subsequently refined by selection into an adaptive phenotype. A critical, but largely untested prediction of plasticity-led evolution (and evolution by natural selection generally) is that the rate and magnitude of evolutionary change should be positively associated with a phenotype's frequency of expression in a population. Essentially, the more often a phenotype is expressed and exposed to selection, the greater its opportunity for adaptive refinement. We tested this prediction by competing against each other spadefoot toad tadpoles from different natural populations that vary in how frequently they express a novel, environmentally induced carnivore ecomorph. As expected, laboratory-reared tadpoles whose parents were derived from populations that express the carnivore ecomorph more frequently were superior competitors for the resource for which this ecomorph is specialized-fairy shrimp. These tadpoles were better at using this resource both because they were more efficient at capturing and consuming shrimp and because they produced more exaggerated carnivore traits. Moreover, they exhibited these more carnivore-like features even without experiencing the inducing cue, suggesting that this ecomorph has undergone an extreme form of plasticity-led evolution-genetic assimilation. Thus, our findings provide evidence that the frequency of trait expression drives the magnitude of adaptive refinement, thereby validating a key prediction of plasticity-led evolution specifically and adaptive evolution generally.

Community disassembly and disease: realistic-but not randomized-biodiversity losses enhance parasite transmission.

Debates over the relationship between biodiversity and disease dynamics underscore the need for a more mechanistic understanding of how changes in host community composition influence parasite transmission. Focusing on interactions between larval amphibians and trematode parasites, we experimentally contrasted the effects of host richness and species composition to identify the individual and joint contributions of both parameters on the infection levels of three trematode species. By combining experimental approaches with field surveys from 147 ponds, we further evaluated how richness effects differed between randomized and realistic patterns of species loss (i.e. community disassembly). Our results indicated that community-level changes in infection levels were owing to host species composition, rather than richness. However, when composition patterns mirrored empirical observations along a natural assembly gradient, each added host species reduced infection success by 12-55%. No such effects occurred when assemblages were randomized. Mechanistically, these patterns were due to non-random host species assembly/disassembly: while highly competent species predominated in low diversity systems, less susceptible hosts became progressively more common as richness increased. These findings highlight the potential for combining information on host traits and assembly patterns to forecast diversity-mediated changes in multi-host disease systems.

Differences in responsiveness and sensitivity to exogenous disruptors of the thyroid gland in three anuran species.

Anuran larval development comprises tissues/organs/systems that are: exclusively of larvae, able to be remodelled, and those of postmetamorphic stages. Also, the anuran larval development is characterized by inter-related parameters: time, size and shape forming part of growth and differentiation. The anuran metamorphosis starts when growth and differentiation achieve a threshold that differs among species since it is regulated by a number of external (environmental) and internal (hormonal) processes. Here we explore the consequences of exogenous disruptors on the thyroid gland (e.g., methimazole and thyroxine as T4) of three species by immersing premetamorphic tadpoles in predetermined concentrations of the disruptors for short periods (10 or 16 days). The species were Pleurodema borellii, Leptodactylus chaquensis, and Dermatonotus muelleri, which all breed in small temporary ponds during the summer, but differ in their ecomorphology. The experiments were conducted to evaluate the effects of these substances on larval development (based in Gosner larval stages), morphometric variation in body parameters (snout-vent and total length by larval stages), and thyroid gland histopathology at the end of the assays. In P. borelli and L. chaquensis, methimazole produces significant increment of size measurements (nonparametric Kruskal-Wallis, p < .05) during stages of digit differentiation and induced thyroid gland hypertrophy. In the three species, T4 exposure accelerated limb development and caused atrophy of thyroid gland. Prolonged T4 exposure in L. chaquensis and D. muelleri triggered metamorphic transformation in the gut and skull cartilages. Discussion about interspecific differences in responsiveness and sensitivity elucidates the importance of hormonal signals to morphological evolution.

How do red-eyed treefrog embryos sense motion in predator attacks? Assessing the role of vestibular mechanoreception.

The widespread ability to alter timing of hatching in response to environmental cues can serve as a defense against threats to eggs. Arboreal embryos of red-eyed treefrogs, Agalychnis callidryas, can hatch up to 30% prematurely to escape predation. This escape-hatching response is cued by physical disturbance of eggs during attacks, including vibrations or motion, and thus depends critically on mechanosensory ability. Predator-induced hatching appears later in development than flooding-induced, hypoxia-cued hatching; thus, its onset is not constrained by the development of hatching ability. It may, instead, reflect the development of mechanosensor function. We hypothesize that vestibular mechanoreception mediates escape-hatching in snake attacks, and that the developmental period when hatching-competent embryos fail to flee from snakes reflects a sensory constraint. We assessed the ontogenetic congruence of escape-hatching responses and an indicator of vestibular function, the vestibulo-ocular reflex (VOR), in three ways. First, we measured VOR in two developmental series of embryos 3-7 days old to compare with the published ontogeny of escape success in attacks. Second, during the period of greatest variation in VOR and escape success, we compared hatching responses and VOR across sibships. Finally, in developmental series, we compared the response of individual embryos to a simulated attack cue with their VOR. The onset of VOR and hatching responses were largely concurrent at all three scales. Moreover, latency to hatch in simulated attacks decreased with increasing VOR. These results are consistent with a key role of the vestibular system in the escape-hatching response of A. callidryas embryos to attacks.

Metabolic switch in energy metabolism mediates the sublethal effects induced by glyphosate-based herbicide on tadpoles of a farmland frog Microhyla fissipes.

Glyphosate-based herbicides (GBHs) are widely-used agricultural chemicals, bringing potential detriments to aquatic organisms. Currently, our understanding of sublethal effects and underlying toxicologic mechanisms of GBHs are still limited, especially in amphibians. Here, the sublethal effects of a commercial GBH (KISSUN®) on tadpoles of a farmland dwelling frog, Microhyla fissipes, were investigated. The 10-d LC50 of "KISSUN®" GBH was 77.5 mg/L. Tadpoles exposed to 60-120 mg/L showed increased preference to higher temperature. After 10 days exposure, obvious growth suppression was observed in survived GBH-stressed tadpoles, characterized by dosage depended decrement in body mass, body width, total length, etc. GBH-stressed tadpoles also showed decreased tail length/snout-vent length ratio and smaller tail muscle fiber diameter. Comparative transcriptomics (control, 60 mg/L and 90 mg/L groups) was conducted to analyze the underlying molecular processes. GBH-stressed tadpoles showed downregulated transcription of ribosomal proteins and cytoskeleton proteins, which could explain their suppressed whole body and tail muscle growth. Moreover, GBH-stressed tadpoles showed transcriptional downregulation of carbohydrate and lipid catabolism, but upregulation of amino acid catabolism. It suggested a metabolic switch from carbohydrate and lipid to amino acid in these tadpoles. Accordingly, there was a trade-off between protein synthesis and energy production in respect to amino acid allocation, and it provided a metabolic explanation for why protein synthesis was downregulated and growth was suppressed in GBH-stressed tadpoles. In combination with existing literatures, we speculated that GBH might directly target the enzymes in carbohydrate and lipid catabolism, and this metabolic effect of GBH might be common to fish and amphibians. In conclusion, our study provided a systematic insight into the sublethal symptoms of GBH-stressed tadpoles, and a metabolic switch from carbohydrate and lipid to amino acid likely underlay some common toxic symptoms of GBHs on both fish and tadpoles.