Search

The question of what the ancient life cycle of tetrapods was like forms a key component in understanding the origin of land vertebrates. The existence of distinct larval forms, as exemplified by many lissamphibians, and their transformation into adults is an important aspect in this field. The temnospondyls, the largest clade of Palaeozoic-Mesozoic non-amniote tetrapods, covered a wide ecomorphological range from fully aquatic to terrestrial taxa. In various species, rich ontogenetic data have accumulated over the past 130 years, permitting the study of early phases of temnospondyl development. In temnospondyls, eight ontogenetic phases have been identified in which the skeleton formed. In branchiosaurids and the eryopiform Sclerocephalus, large parts of the ossification sequence are now known. Most taxa in which small specimens are preserved had aquatic larvae with external gills that superficially resemble larval salamanders. In the edopoids, dvinosaurs, and eryopiforms, the larvae developed slowly, with incompletely ossified axial and appendicular skeletons, but possessed a fast-developing dermal skull with strong teeth. Irrespective of adult terrestriality or a fully aquatic life, there was no drastic transformation during later ontogeny, but a slow and steady acquisition of adult features. In dissorophoids, the limbs developed at a much faster pace, whereas skull formation was slowed down, especially in the amphibamiforms, and culminating in the neotenic Branchiosauridae. In the zatracheid Acanthostomatops, slow but profound transformation led to a fully terrestrial adult. The basal dissorophoid Stegops retained rapid development of dermal skull bones and established a fully dentigerous, strongly ossified palate early. In Micromelerpeton, formation of the last skull bones was slightly delayed and metamorphosis remained a long and steady phase of morphological transformations. In amphibamiforms, metamorphosis became more drastic, with an increasing number of events packed into a short phase of ontogeny. This is exemplified by Apateon, Platyrhinops, and Amphibamus in which this condensation was maximised. We distinguish three different types of metamorphosis (morphological, ecological and drastic) that evolved cumulatively in early tetrapods and within temnospondyls., (© 2024 The Authors. Biological Reviews published by John Wiley & Sons Ltd on behalf of Cambridge Philosophical Society.)

Initiatives to protect 30% of Earth by 2030 prompt evaluation of how to efficiently target shortcomings in the global protected area (PA) network. Focusing on amphibians, the most vulnerable vertebrate class, we illustrate the conservation value of microreserves, a term we employ here to refer to reserves of <10 km 2 . We report that the network continues to under-represent threatened amphibians and that, despite this clear shortcoming in land-based conservation, the creation of PAs protecting amphibians slowed after 2010. By proving something previously assumed-that amphibians generally have smaller ranges than other terrestrial vertebrates-we demonstrate that microreserves could protect a substantial portion of many amphibian ranges, particularly threatened species. We find existing microreserves are capable of hosting an amphibian species richness similar to PAs 1000-10,00X larger, and we show that amphibians' high beta diversity means that microreserves added to a growing PA network cover amphibian species 1.5-6x faster than larger size categories. We propose that stemming global biodiversity loss requires that we seriously consider the conservation potential of microreserves, using them to capture small-range endemics that may otherwise be omitted from the PA network entirely., (© 2024. The Author(s).)

Among human actions threatening biodiversity, the release of anthropogenic chemical pollutants which have become ubiquitous in the environment, is a major concern. Chemical pollution can induce damage to macromolecules by causing the overproduction of reactive oxygen species, affecting the redox balance of animals. In species undergoing metamorphosis (i.e. the vast majority of the extant animal species), antioxidant responses to chemical pollution may differ between pre- and post-metamorphic stages. Here, we meta-analysed ( N  = 104 studies, k =  2283 estimates) the impact of chemical pollution on redox balance across the three major amphibian life stages (embryo, tadpole, adult). Before metamorphosis, embryos did not experience any redox change while tadpoles activate their antioxidant pathways and do not show increased oxidative damage from pollutants. Tadpoles may have evolved stronger defences against pollutants to reach post-metamorphic life stages. In contrast, post-metamorphic individuals show only weak antioxidant responses and marked oxidative damage in lipids. The type of pollutant (i.e. organic versus inorganic) has contrasting effects across amphibian life stages. Our findings show a divergent evolution of the redox balance in response to pollutants across life transitions of metamorphosing amphibians, most probably a consequence of differences in the ecological and developmental processes of each life stage.

This study presents a comprehensive analysis of a two-patch, two-life stage SI model without recovery from infection, focusing on the dynamics of disease spread and host population viability in natural populations. The model, inspired by real-world ecological crises like the decline of amphibian populations due to chytridiomycosis and sea star populations due to Sea Star Wasting Disease, aims to understand the conditions under which a sink host population can present ecological rescue from a healthier, source population. Mathematical and numerical analyses reveal the critical roles of the basic reproductive numbers of the source and sink populations, the maturation rate, and the dispersal rate of juveniles in determining population outcomes. The study identifies basic reproduction numbers R 0 for each of the patches, and conditions for the basic reproduction numbers to produce a receiving patch under which its population. These findings provide insights into managing natural populations affected by disease, with implications for conservation strategies, such as the importance of maintaining reproductively viable refuge populations and considering the effects of dispersal and maturation rates on population recovery. The research underscores the complexity of host-pathogen dynamics in spatially structured environments and highlights the need for multi-faceted approaches to biodiversity conservation in the face of emerging diseases., (© 2024. The Author(s), under exclusive licence to the Society for Mathematical Biology.)

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

Amphibians and fishes play a central role in shaping the structure and function of freshwater environments. These organisms have a limited capacity to disperse across different habitats and the thermal buffer offered by freshwater systems is small. Understanding determinants and patterns of their physiological sensitivity across life history is, therefore, imperative to predicting the impacts of climate change in freshwater systems. Based on a systematic literature review including 345 experiments with 998 estimates on 96 amphibian (Anura/Caudata) and 93 freshwater fish species (Teleostei), we conducted a quantitative synthesis to explore phylogenetic, ontogenetic, and biogeographic (thermal adaptation) patterns in upper thermal tolerance (CT max ) and thermal acclimation capacity (acclimation response ratio, ARR) as well as the influence of the methodology used to assess these thermal traits using a conditional inference tree analysis. We found globally consistent patterns in CT max and ARR, with phylogeny (taxa/order), experimental methodology, climatic origin, and life stage as significant determinants of thermal traits. The analysis demonstrated that CT max does not primarily depend on the climatic origin but on experimental acclimation temperature and duration, and life stage. Higher acclimation temperatures and longer acclimation times led to higher CT max values, whereby Anuran larvae revealed a higher CT max than older life stages. The ARR of freshwater fishes was more than twice that of amphibians. Differences in ARR between life stages were not significant. In addition to phylogenetic differences, we found that ARR also depended on acclimation duration, ramping rate, and adaptation to local temperature variability. However, the amount of data on early life stages is too small, methodologically inconsistent, and phylogenetically unbalanced to identify potential life cycle bottlenecks in thermal traits. We, therefore, propose methods to improve the robustness and comparability of CT max /ARR data across species and life stages, which is crucial for the conservation of freshwater biodiversity under climate change., (© 2024 The Authors. Global Change Biology published by John Wiley & Sons Ltd.)

The trade-off between offspring size and number is central to life history strategies. Both the evolutionary gain of parental care or more favorable habitats for offspring development are predicted to result in fewer, larger offspring. However, despite much research, it remains unclear whether and how different forms of care and habitats drive the evolution of the trade-off. Using data for over 800 amphibian species, we demonstrate that, after controlling for allometry, amphibians with direct development and those that lay eggs in terrestrial environments have larger eggs and smaller clutches, while different care behaviors and adaptations vary in their effects on the trade-off. Specifically, among the 11 care forms we considered at the egg, tadpole and juvenile stage, egg brooding, male egg attendance, and female egg attendance increase egg size; female tadpole attendance and tadpole feeding decrease egg size, while egg brooding, tadpole feeding, male tadpole attendance, and male tadpole transport decrease clutch size. Unlike egg size that shows exceptionally high rates of phenotypic change in just 19 branches of the amphibian phylogeny, clutch size has evolved at exceptionally high rates in 135 branches, indicating episodes of strong selection; egg and tadpole environment, direct development, egg brooding, tadpole feeding, male tadpole attendance, and tadpole transport explain 80% of these events. By explicitly considering diversity in parental care and offspring habitat by stage of offspring development, this study demonstrates that more favorable conditions for offspring development promote the evolution of larger offspring in smaller broods and reveals that the diversity of parental care forms influences the trade-off in more nuanced ways than previously appreciated., Competing Interests: The authors have declared that no competing interests exist.

The chytrid fungal pathogens Batrachochytrium dendrobatidis and Batrachochytrium salamandrivorans cause the skin disease chytridiomycosis in amphibians, which is driving a substantial proportion of an entire vertebrate class to extinction. Mitigation of its impact is largely unsuccessful and requires a thorough understanding of the mechanisms underpinning the disease ecology. By identifying skin factors that mediate key events during the early interaction with B. salamandrivorans zoospores, we discovered a marker for host colonization. Amphibian skin associated beta-galactose mediated fungal chemotaxis and adhesion to the skin and initiated a virulent fungal response. Fungal colonization correlated with the skin glycosylation pattern, with cutaneous galactose content effectively predicting variation in host susceptibility to fungal colonization between amphibian species. Ontogenetic galactose patterns correlated with low level and asymptomatic infections in salamander larvae that were carried over through metamorphosis, resulting in juvenile mortality. Pronounced variation of galactose content within some, but not all species, may promote the selection for more colonization resistant host lineages, opening new avenues for disease mitigation., (© 2021. The Author(s).)

Amphibians display very diverse life cycles and development can be direct, where it occurs in ovo and a juvenile hatches directly, or biphasic, where an aquatic larva hatches and later undergoes metamorphosis followed by sexual maturation. In both cases, metamorphosis, corresponds to the post embryonic transition (PETr). A third strategy, only found in Urodeles, is more complex as larvae reach sexual maturity before metamorphosis, which can become accessory. The resulting paedomorphs retain their larval characters and keep their aquatic habitat. Does it mean that paedomorphs do not undergo PETr? Recent work using high throughput technologies coupled to system biology and developmental endocrinology revisited this question and provided novel datasets indicating that a paedomorph's "larval" tissue undergoes a proper developmental transition. Together with historical data, we propose that this transition is a marker of the PETr, which would be distinct from metamorphosis. This implies that (a) complex life cycles would result from the uncoupling of PETr and metamorphosis, and (b) biphasic life cycles would be a special cases where they occur simultaneously., (© 2021 American Association of Anatomists.)

Sex is determined genetically in amphibians; however, little is known about the sex chromosomes, testis-determining genes, and the genes involved in testis differentiation in this class. Certain inherent characteristics of the species of this group, like the homomorphic sex chromosomes, the high diversity of the sex-determining mechanisms, or the existence of polyploids, may hinder the design of experiments when studying how the gonads can differentiate. Even so, other features, like their external development or the possibility of inducing sex reversal by external treatments, can be helpful. This review summarizes the current knowledge on amphibian sex determination, gonadal development, and testis differentiation. The analysis of this information, compared with the information available for other vertebrate groups, allows us to identify the evolutionarily conserved and divergent pathways involved in testis differentiation. Overall, the data confirm the previous observations in other vertebrates-the morphology of the adult testis is similar across different groups; however, the male-determining signal and the genetic networks involved in testis differentiation are not evolutionarily conserved.

In this review article, topics of the embryonic origin of the adenohypophysis and hypothalamus and the development of the hypothalamo-hypophyseal system for the completion of metamorphosis in amphibians are included. The primordium of the adenohypophysis as well as the primordium of the hypothalamus in amphibians is of neural origin as shown in other vertebrates, and both are closely associated with each other at the earliest stage of development. Metamorphosis progresses via the interaction of thyroid hormone and adrenal corticosteroids, of which secretion is enhanced by thyrotropin and corticotropin, respectively. However, unlike in mammals, the hypothalamic releasing factor for thyrotropin is not thyrotropin-releasing hormone (TRH), but corticotropin-releasing factor (CRF) and the major releasing factor for corticotropin is arginine vasotocin (AVT). Prolactin, the release of which is profoundly enhanced by TRH at the metamorphic climax, is another pituitary hormone involved in metamorphosis. Prolactin has a dual role: modulation of the metamorphic speed and the development of organs for adult life. The secretory activities of the pituitary cells containing the three above-mentioned pituitary hormones are elevated toward the metamorphic climax in parallel with the activities of the CRF, AVT, and TRH neurons., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Alternative splicing is widespread throughout eukaryotic genomes and greatly increases transcriptomic diversity. Many alternative isoforms have functional roles in developmental processes and are precisely temporally regulated. To facilitate the study of alternative splicing in a developmental context, we created MeDAS, a Metazoan Developmental Alternative Splicing database. MeDAS is an added-value resource that re-analyses publicly archived RNA-seq libraries to provide quantitative data on alternative splicing events as they vary across the time course of development. It has broad temporal and taxonomic scope and is intended to assist the user in identifying trends in alternative splicing throughout development. To create MeDAS, we re-analysed a curated set of 2232 Illumina polyA+ RNA-seq libraries that chart detailed time courses of embryonic and post-natal development across 18 species with a taxonomic range spanning the major metazoan lineages from Caenorhabditis elegans to human. MeDAS is freely available at https://das.chenlulab.com both as raw data tables and as an interactive browser allowing searches by species, tissue, or genomic feature (gene, transcript or exon ID and sequence). Results will provide details on alternative splicing events identified for the queried feature and can be visualised at the gene-, transcript- and exon-level as time courses of expression and inclusion levels, respectively., (© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Research on the thermal ecology and physiology of free-living organisms is accelerating as scientists and managers recognize the urgency of the global biodiversity crisis brought on by climate change. As ectotherms, temperature fundamentally affects most aspects of the lives of amphibians and reptiles, making them excellent models for studying how animals are impacted by changing temperatures. As research on this group of organisms accelerates, it is essential to maintain consistent and optimal methodology so that results can be compared across groups and over time. This review addresses the utility of reptiles and amphibians as model organisms for thermal studies by reviewing the best practices for research on their thermal ecology and physiology, and by highlighting key studies that have advanced the field with new and improved methods. We end by presenting several areas where reptiles and amphibians show great promise for further advancing our understanding of how temperature relations between organisms and their environments are impacted by global climate change., (© 2020 Wiley Periodicals LLC.)

Infectious diseases are considered major threats to biodiversity, however strategies to mitigate their impacts in the natural world are scarce and largely unsuccessful. Chytridiomycosis is responsible for the decline of hundreds of amphibian species worldwide, but an effective disease management strategy that could be applied across natural habitats is still lacking. In general amphibian larvae can be easily captured, offering opportunities to ascertain the impact of altering the abundance of hosts, considered to be a key parameter affecting the severity of the disease. Here, we report the results of two experiments to investigate how altering host abundance affects infection intensity in amphibian populations of a montane area of Central Spain suffering from lethal amphibian chytridiomycosis. Our laboratory-based experiment supported the conclusion that varying density had a significant effect on infection intensity when salamander larvae were housed at low densities. Our field experiment showed that reducing the abundance of salamander larvae in the field also had a significant, but weak, impact on infection the following year, but only when removals were extreme. While this suggests adjusting host abundance as a mitigation strategy to reduce infection intensity could be useful, our evidence suggests only heavy culling efforts will succeed, which may run contrary to objectives for conservation., Competing Interests: The authors have declared that no competing interests exist.

Landscape heterogeneity and fragmentation are key challenges for biodiversity conservation. As Earth's landscape is increasingly dominated by anthropogenic land use, it is clear that broad-scale systems of nature reserves connected by corridors are needed to enable the dispersal of flora and fauna. The European Union currently supports a continent-wide network of protected areas, the Natura 2000 program, but this program lacks the necessary connectivity component. To examine whether a comprehensive network could be built in order to protect amphibians and reptiles, two taxonomic groups sensitive to environmental changes due to their physiological constrains and low dispersal capacity, we used species' distribution maps, the sites of community interest (SCIs) in Romania, and landscape resistance rasters. Except Vipera ursinii rakosiensis, all amphibians and reptiles had corridors mapped that, when assembled, provided linkages for up to 27 species. Natura 2000 species were not good candidates for umbrella species as these linkages covered only 17% of the corridors for all species. Important Areas for Connectivity were identified in the Carpathian Mountains and along the Danube River, further confirming these regions as hot spots for biodiversity in Europe, where successful linkages are most likely. In the end, while such corridors may not be created just for amphibians and reptiles, they can easily be incorporated into more complex linkages with corridors for more charismatic species, therefore enhancing the corridors' value in terms of quality and structure.

Two species of parasitic fungi from the phylum Chytridiomycota (chytrids) are annihilating global amphibian populations. These chytrid species-Batrachochytrium dendrobatidis and B. salamandrivorans-have high rates of mortality and transmission. Upon establishing infection in amphibians, chytrids rapidly multiply within the skin and disrupt their hosts' vital homeostasis mechanisms. Current disease models suggest that chytrid fungi locate and infect their hosts during a motile, unicellular 'zoospore' life stage. Moreover, other chytrid species parasitize organisms from across the tree of life, making future epidemics in new hosts a likely possibility. Efforts to mitigate the damage and spread of chytrid disease have been stymied by the lack of knowledge about basic chytrid biology and tools with which to test molecular hypotheses about disease mechanisms. To overcome this bottleneck, we have developed high-efficiency delivery of molecular payloads into chytrid zoospores using electroporation. Our electroporation protocols result in payload delivery to between 75 and 97% of living cells of three species: B. dendrobatidis, B. salamandrivorans, and a non-pathogenic relative, Spizellomyces punctatus. This method lays the foundation for molecular genetic tools needed to establish ecological mitigation strategies and answer broader questions in evolutionary and cell biology.

Although metamorphosis is widespread in the animal kingdom, several species have evolved life-cycle modifications to avoid complete metamorphosis. Some species, for example, many salamanders and newts, have deleted the adult stage via a process called paedomorphosis. Others, for example, some frog species and marine invertebrates, no longer have a distinct larval stage and reach maturation via direct development. Here we study which ecological conditions can lead to the loss of metamorphosis via the evolution of direct development. To do so, we use size-structured consumer-resource models in conjunction with the adaptive-dynamics approach. In case the larval habitat deteriorates, individuals will produce larger offspring and in concert accelerate metamorphosis. Although this leads to the evolutionary transition from metamorphosis to direct development when the adult habitat is highly favorable, the population will go extinct in case the adult habitat does not provide sufficient food to escape metamorphosis. With a phylogenetic approach we furthermore show that among amphibians the transition of metamorphosis to direct development is indeed, in line with model predictions, conditional on and preceded by the evolution of larger egg sizes., (© 2020 The Authors. Evolution published by Wiley Periodicals LLC on behalf of The Society for the Study of Evolution.)

Size is a fundamental feature of biology that affects physiology at all levels, from the organism to organs and tissues to cells and subcellular structures. How size is determined at these different levels, and how biological structures scale to fit together and function properly are important open questions. Historically, amphibian systems have been extremely valuable to describe scaling phenomena, as they occupy some of the extremes in biological size and are amenable to manipulations that alter genome and cell size. More recently, the application of biochemical, biophysical, and embryological techniques to amphibians has provided insight into the molecular mechanisms underlying scaling of subcellular structures to cell size, as well as how perturbation of normal size scaling impacts other aspects of cell and organism physiology., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Most predictions of how populations and species of ectotherms will respond to global warming are based on estimates of the temperature at which organisms lose motor control (i.e., CT max - the Critical Thermal Maximum). Here, we describe a non-lethal protocol and ethograms to estimate the relative tolerance of amphibians to increasing temperatures. These methods-suitable for field or laboratory conditions-are replicable, inexpensive and applicable to both post-metamorphic stages and organisms with direct development. We illustrate the use of this standardized protocol for four amphibians from a tropical cloud forest in Veracruz, Mexico with contrasting life histories: a lungless salamander (Aquiloeurycea cafetalera: Plethodontidae), a leaf-litter frog (Craugastor rhodopis: Craugastoridae), a semiaquatic frog (Lithobates berlandieri: Ranidae), and a tree frog (Rheohyla miotympanum: Hylidae). We identified four behavioral responses preceding CT max for all amphibians included in this study: 1) Optimal Activity Range, 2) Supra-optimal Activity Range, 3) Heat Stress Range, and 4) Involuntary Movements Range. Additionally, we identified a fifth parameter associated with resilience to heat shock: 5) Recovery Stage after reaching CT max . We conclude that the behavioral responses preceding the Critical Thermal Maximum are as informative as CT max . Using behavioral responses to estimate thermal tolerance has the additional advantage of reducing the risk of injury or death of amphibians during physiological experiments., Competing Interests: Declaration of competing interest No competing interests declared., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The hypothesis here presented tries to explain why organ regeneration is present in fish and amphibians (anamniotes) but is absent in reptiles, birds and mammals (amniotes). Anamniotes possess complex life cycles including larvae and metamorphosis stages, the latter representing a physiological form of organ destruction and regeneration coded in their genome that can be reactivated in adults in the form of regeneration. Part of the genome for larvae and metamorphosis phases was likely lost in amniotes with the evolution of direct development, the potentiation of the immune system and the increase in complexity of the nervous system. These events consequently determined incapability for organ regeneration in extant amniotes with the exception of the lizard tail. This likely derives from the evolution of a mechanism of immunosuppression that allows the regeneration of the tail although the complete morphogenetic plane of tail embryogenesis is lost. The lizard model of imperfect but outstanding organ regeneration indicates the possibility to improve organ regeneration also in other amniotes. In fact, the induction of a blastema in the amputated lizard limb has stimulated the formation of short limbs containing cartilaginous bones of the femur, tibia and fibula, and these experiments foster some hope for future attempts to induce limb and digit regeneration also in mammals.

It is a widely held belief that environmental contaminants contribute to the decline of amphibian populations. By spending most of their early life in water and later stages on the land, amphibians face a constant risk of exposure to pesticides and other chemical pollutants in both aquatic and terrestrial environments. This review presents an overview of the studies carried out in Italian amphibians to highlight hazardous effects of bioaccumulation of chemical pollutants in juveniles and adults in various contaminated environments. Further, the studies in the laboratory setting assessing the effects of chemical pollutants on reproductive and developmental processes are reported. These studies and their relative references have been summarized in a tabular form. Three prominent contaminant groups were identified: herbicides, insecticides, and fungicides; and only a few works reported the effects of other chemical pollutants. Each pollutant group has been delegated to a section. All through the literature survey, it is seen that interest in this topic in Italy is very recent and sparse, where only a few anuran and caudata species and only some chemical pollutants have been studied., (© 2019 Wiley Periodicals, Inc.)

In 1901 T.H. Morgan proposed in "Regeneration" that pattern formation in amphibian limb regeneration is a stepwise process. Since, biologist have continued to piece together the molecular components of this process to better understand the "patterning code" responsible for regenerate formation. Within this context, several different models have been proposed; however, all are based on one of two underlying hypotheses. The first is the "morphogen hypothesis" that dictates that pattern emerges from localized expression of signaling molecules, which produce differing position-specific cellular responses in receptive cells depending on the intensity of the signal. The second hypothesis is that cells in the remaining tissues retain memory of their patterning information, and use this information to generate new cells with the missing positional identities. A growing body of evidence supports the possibility that these two mechanisms are not mutually exclusive. Here, we propose our theory of hierarchical pattern formation, which consists of 4 basic steps. The first is the existence of cells with positional memory. The second is the communication of positional information through cell-cell interactions in a regeneration-permissive environment. The third step is the induction of molecular signaling centers. And the last step is the interpretation of these signals by specialized cell types to ultimately restore the limb in its entirety. Biological codes are intertwined throughout this model, and we will discuss their multiple roles and mechanisms., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Many animals undergo a transition during their lifetime from a larval to an adult form, a major developmental change known as metamorphosis. This developmental process, which involves behavioural, morphological, physiological and biochemical changes, has a broad phylogenetic distribution, occurring in diverse branches of the animal kingdom, from invertebrates (molluscs, arthropods, tunicates) to certain classes of vertebrates, including amphibians (Figure 1). This phenomenon, which has fascinated biologists for centuries, remains an attractive experimental model for studying mechanisms of post-embryonic development as well as molecular mechanisms underlying hormonal regulation., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

A growing number of biologists have begun asking whether environmentally induced phenotypic change--'phenotypic plasticity'--precedes and facilitates the origin and canalization of novel, complex phenotypes. However, such 'plasticity-first evolution' (PFE) remains controversial. Here, we summarize the PFE hypothesis and describe how it can be evaluated in natural systems. We then review the evidence for PFE from amphibians (a group in which phenotypic plasticity is especially widespread) and describe how phenotypic plasticity might have facilitated macroevolutionary change. Finally, we discuss what is known about the proximate mechanisms of PFE in amphibians. We close with suggestions for future research. As we describe, amphibians offer some of the best support for plasticity's role in the origin of evolutionary novelties., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The exponential increase in species introductions during the Anthropocene has brought about a major loss of biodiversity. Amphibians have suffered large declines, with more than 16% considered to be threatened by invasive species. We conducted a global meta-analysis of the impacts of alien species on native amphibians to determine which aspects of amphibian ecology are most affected by plant, invertebrate, fish, amphibian, reptile, or mammal introductions. Measures of fitness were most strongly affected; amphibian performance was consistently lower in the presence of alien species. While exposure to alien species caused a significant decrease in amphibian behavioural activity when compared with a no species control, this response was stronger towards a control of native impacting species. This indicates a high degree of prey naiveté towards alien species and highlights the importance of using different types of controls in empirical studies. Alien invertebrates had the greatest overall impact on amphibians. This study sets a new agenda for research on biological invasions, highlighting the lack of studies investigating the impacts of alien species on amphibian terrestrial life-history stages. It also emphasizes the strong ecological impacts that alien species have on amphibian fitness and suggests that future introductions or global spread of alien invertebrates could strongly exacerbate current amphibian declines.

Background Endochondral ossification, the process by which most of the skeleton is formed, is accurately regulated by many specific groups of molecules and extracellular matrix components. The molecular mechanisms of endochondral ossification have been extensively investigated in mammals. However, there are few studies about it in amphibians. Results Ossification of femur and tibiofibula was observed at Gs 40, and tarsals and metatarsals were ossified at Gs 42. Most of the skeletons in hindlimbs were completely ossified except for articular cartilages at Gs 46. Thirty-two genes related to endochondral ossification were found in the differentially expressed genes (DEGs) library of hindlimbs, and nine of these genes were validated by qRT-PCR during metamorphosis. Sox9 was expressed in the columnar, prehypertrophic, and upper hypertrophic zones, and the expression of Ihh was observed in prehypertrophic chondrocyte zone in hindlimbs of B. gargarizans. Conclusions The ossification of hindlimbs increased gradually, and the ossification sequence was from proximal to distal in B. gargarizans during metamorphosis. Thirty-two genes found in the DEGs library were related to the regulation of endochondral ossification of hindlimbs in amphibians. The present study will provide a valuable genomic resource for the future study of endochondral ossification in amphibian. Developmental Dynamics 247:1121-1134, 2018. © 2018 Wiley Periodicals, Inc., (© 2018 Wiley Periodicals, Inc.)

It is theoretically possible to protect large fractions of species in relatively small regions. For plants, 85% of species occur entirely within just over a third of the Earth's land surface, carefully optimized to maximize the species captured. Well-known vertebrate taxa show similar patterns. Protecting half of Earth might not be necessary, but would it be sufficient given the current trends of protection? The predilection of national governments is to protect areas that are "wild," that is, typically remote, cold, or arid. Unfortunately, those areas often hold relatively few species. Wild places likely afford the easier opportunities for the future expansion of protected areas, with the expansion into human-dominated landscapes the greater challenge. We identify regions that are not currently protected, but that are wild, and consider which of them hold substantial numbers of especially small-ranged vertebrate species. We assess how successful the strategy of protecting the wilder half of Earth might be in conserving biodiversity. It is far from sufficient. (Protecting large wild places for reasons other than biodiversity protection, such as carbon sequestration and other ecosystem services, might still have importance.) Unexpectedly, we also show that, despite the bias in establishing large protected areas in wild places to date, numerous small protected areas are in biodiverse places. They at least partially protect significant fractions of especially small-ranged species. So, while a preoccupation with protecting large areas for the sake of getting half of Earth might achieve little for biodiversity, there is more progress in protecting high-biodiversity areas than currently appreciated. Continuing to prioritize the right parts of Earth, not just the total area protected, is what matters for biodiversity.

The amphibian Xenopus laevis has long been used as a model for studying vertebrate cell and developmental biology largely due to the easiness to manipulate this system in vivo and in vitro. While most of the developmental studies have been on Xenopus embryogenesis, considerable efforts have been made to understand its metamorphosis, a process mimicking postembryonic development in mammals when many organs mature into their adult forms in the presence of high levels of thyroid hormone (T3). Amphibian metamorphosis is totally dependent on T3 and offers a number of advantages for experimental analyses compared to the late stage, uterus-enclosed mammalian embryos. Earlier studies on metamorphosis in Xenopus laevis have revealed dual functions of T3 receptors (TR) during premetamorphic development and metamorphosis as well as important roles of TR-interacting corepressors and coactivators during these two periods, respectively. The development of gene-editing technologies that functions in amphibians in recent years has made possible for the first time to study function of endogenous TRs, especially in the highly related diploid anuran species Xenopus tropicalis. Here, we first review the current mechanistic understanding of the regulation of metamorphosis by T3 and TR, and then describe a detailed method to use TALEN to knock out TRα for studying its role in gene regulation by T3 in vivo and Xenopus development.

The gastrointestinal tract is a well-characterized target of thyroid hormones and thyroid hormone nuclear receptors TRs, as extensively described in the literature. The paradigm is its important remodelling in amphibians during thyroid hormone-dependent metamorphosis. Interestingly, several studies have described the conservation of this hormonal signal during intestinal development in mammals. Additional data suggested that it may also play a role in intestinal homeostasis, stem cell physiology and progenitor commitment as well as in tumour development. It is worth underlining that in the mammalian intestine the functionality of the TRα1 receptor is coordinated and integrated with other signalling pathways, such as Wnt and Notch, specifically at the level of stem/progenitor cell populations. Here, we summarize these data and concepts and discuss this new role for thyroid hormones and the TRα1 receptor in the biology of intestinal epithelial precursor cells., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Chytridiomycosis, due to the fungus Batrachochytrium dendrobatidis (Bd), has been associated with the alarming decline and extinction crisis of amphibians worldwide. Because conservation programs are implemented locally, it is essential to understand how the complex interactions among host species, climate and human activities contribute to Bd occurrence at regional scales. Using weighted phylogenetic regressions and model selection, we investigated geographic patterns of Bd occurrence along a latitudinal gradient of 1500 km within a biodiversity hot spot in Chile (1845 individuals sampled from 253 sites and representing 24 species), and its association with climatic, socio-demographic and economic variables. Analyses show that Bd prevalence decreases with latitude although it has increased by almost 10% between 2008 and 2013, possibly reflecting an ongoing spread of Bd following the introduction of Xenopus laevis. Occurrence of Bd was higher in regions with high gross domestic product (particularly near developed centers) and with a high variability in rainfall regimes, whereas models including other bioclimatic or geographic variables, including temperature, exhibited substantially lower fit and virtually no support based on Akaike weights. In addition, Bd prevalence exhibited a strong phylogenetic signal, with five species having high numbers of infected individuals and higher prevalence than the average of 13.3% across all species. Taken together, our results highlight that Bd in Chile might still be spreading south, facilitated by a subset of species that seem to play an important epidemiological role maintaining this pathogen in the communities, in combination with climatic and human factors affecting the availability and quality of amphibian breeding sites. This information may be employed to design conservation strategies and mitigate the impacts of Bd in the biodiversity hot spot of southern Chile, and similar studies may prove useful to disentangle the role of different factors contributing to the emergence and spread of this catastrophic disease., (© 2017 John Wiley & Sons Ltd.)

Urea-based herbicides are applied in agriculture to control broadleaf and grassy weeds, acting to either inhibit photosynthesis at photosystem II (phenylureas) or to inhibit acetolactate synthase acetohydroxyacid synthase (sulfonylureas). While there are different chemical formulas for urea-based herbicides, the phenylureas are a widely used class in North America and have been detected in aquatic environments due to agricultural run-off. Here, we summarize the current state of the literature, synthesizing data on phenylureas and their biological effects in two non-target animals, fish and amphibians, with a primary focus on diuron and linuron. In fish, although the acutely lethal effects of diuron in early life stages appear to be >1mg/L, recent studies measuring sub-lethal behavioural and developmental endpoints suggest that diuron causes adverse effects at lower concentrations (i.e. <0.1mg/L). Considerably less toxicity data exist for amphibians, and this is a knowledge gap in the literature. In terms of sub-lethal effects and mode of action (MOA), linuron is well documented to have anti-androgenic effects in vertebrates, including fish. However, there are other MOAs that are not adequately assessed in toxicology studies. In order to identify additional potential MOAs, we conducted in silico analyses for linuron and diuron that were based upon transcriptome studies and chemical structure-function relationships (i.e. ToxCast™, Prediction of Activity Spectra of Substances). Based upon these analyses, we suggest that steroid biosynthesis, cholesterol metabolism and pregnane X receptor activation are common targets, and offer some new endpoints for future investigations of phenylurea herbicides in non-target animals., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Recent expansion of the scale of human activities poses severe threats to Earth's life-support systems. Increasingly, protected areas (PAs) are expected to serve dual goals: protect biodiversity and secure ecosystem services. We report a nationwide assessment for China, quantifying the provision of threatened species habitat and four key regulating services-water retention, soil retention, sandstorm prevention, and carbon sequestration-in nature reserves (the primary category of PAs in China). We find that China's nature reserves serve moderately well for mammals and birds, but not for other major taxa, nor for these key regulating ecosystem services. China's nature reserves encompass 15.1% of the country's land surface. They capture 17.9% and 16.4% of the entire habitat area for threatened mammals and birds, but only 13.1% for plants, 10.0% for amphibians, and 8.5% for reptiles. Nature reserves encompass only 10.2-12.5% of the source areas for the four key regulating services. They are concentrated in western China, whereas much threatened species' habitat and regulating service source areas occur in eastern provinces. Our analysis illuminates a strategy for greatly strengthening PAs, through creating the first comprehensive national park system of China. This would encompass both nature reserves, in which human activities are highly restricted, and a new category of PAs for ecosystem services, in which human activities not impacting key services are permitted. This could close the gap in a politically feasible way. We also propose a new category of PAs globally, for sustaining the provision of ecosystems services and achieving sustainable development goals.

It is well known that the hypothalamic neuropeptide gonadotropin-releasing hormone (GnRH) plays an important role as a primary factor regulating gonadotropin secretion in reproductive processes in vertebrates. The discovery of the presence of a gonadotropin-inhibitory hormone (GnIH) in the brains of birds has further contributed to our understanding of the reproduction control by the brain. GnIH plays a key role in inhibition of reproduction and acts on the pituitary gland and GnRH neurons via a novel G protein-coupled receptor (GPR147). GnIH decreases gonadotropin synthesis and release, thus inhibiting gonadal development and maintenance. The GnRH and GnIH neuronal peptidergic systems are well reported in mammals and birds, but limited information is available regarding their presence and localization in the brains of other vertebrate species, such as reptiles, amphibians and fishes. The aim of this review is to compile and update information on the localization of GnRH and GnIH neuronal systems, with a particular focus on amphibians, summarizing the neuroanatomical distribution of GnIH and GnRH and emphasizing the discovery of GnIH based on RFamide peptides and GnIH orthologous peptides found in other vertebrates and their functional significance., (Copyright © 2016 Elsevier Inc. All rights reserved.)

In vertebrates, sexual differentiation of the reproductive system and brain is tightly orchestrated by organizational and activational effects of endogenous hormones. In mammals and birds, the organizational period is typified by a surge of sex hormones during differentiation of specific neural circuits; whereas activational effects are dependent upon later increases in these same hormones at sexual maturation. Depending on the reproductive organ or brain region, initial programming events may be modulated by androgens or require conversion of androgens to estrogens. The prevailing notion based upon findings in mammalian models is that male brain is sculpted to undergo masculinization and defeminization. In absence of these responses, the female brain develops. While timing of organizational and activational events vary across taxa, there are shared features. Further, exposure of different animal models to environmental chemicals such as xenoestrogens such as bisphenol A-BPA and ethinylestradiol-EE2, gestagens, and thyroid hormone disruptors, broadly classified as neuroendocrine disrupting chemicals (NED), during these critical periods may result in similar alterations in brain structure, function, and consequently, behaviors. Organizational effects of neuroendocrine systems in mammals and birds appear to be permanent, whereas teleost fish neuroendocrine systems exhibit plasticity. While there are fewer NED studies in amphibians and reptiles, data suggest that NED disrupt normal organizational-activational effects of endogenous hormones, although it remains to be determined if these disturbances are reversible. The aim of this review is to examine how various environmental chemicals may interrupt normal organizational and activational events in poikilothermic vertebrates. By altering such processes, these chemicals may affect reproductive health of an animal and result in compromised populations and ecosystem-level effects.

Small heat shock proteins (sHSPs) are a superfamily of molecular chaperones with important roles in protein homeostasis and other cellular functions. Amphibians, reptiles, fish and birds have a shsp gene called hsp30, which was also referred to as hspb11 or hsp25 in some fish and bird species. Hsp30 genes, which are not found in mammals, are transcribed in response to heat shock or other stresses by means of the heat shock factor that is activated in response to an accumulation of unfolded protein. Amino acid sequence analysis revealed that representative HSP30s from different classes of non-mammalian vertebrates were distinct from other sHSPs including HSPB1/HSP27. Studies with amphibian and fish recombinant HSP30 determined that they were molecular chaperones since they inhibited heat- or chemically-induced aggregation of unfolded protein. During non-mammalian vertebrate development, hsp30 genes were differentially expressed in selected tissues. Also, heat shock-induced stage-specific expression of hsp30 genes in frog embryos was regulated at the level of chromatin structure. In adults and/or tissue culture cells, hsp30 gene expression was induced by heat shock, arsenite, cadmium or proteasomal inhibitors, all of which enhanced the production of unfolded/damaged protein. Finally, immunocytochemical analysis of frog and chicken tissue culture cells revealed that proteotoxic stress-induced HSP30 accumulation co-localized with aggresome-like inclusion bodies. The congregation of damaged protein in aggresomes minimizes the toxic effect of aggregated protein dispersed throughout the cell. The current availability of probes to detect the presence of hsp30 mRNA or encoded protein has resulted in the increased use of hsp30 gene expression as a marker of proteotoxic stress in non-mammalian vertebrates., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Vestibulo-ocular reflexes (VOR) ensure gaze stability during locomotion and passively induced head/body movements. In precocial vertebrates such as amphibians, vestibular reflexes are required very early at the onset of locomotor activity. While the formation of inner ears and the assembly of sensory-motor pathways is largely completed soon after hatching, angular and translational/tilt VOR display differential functional onsets and mature with different time courses. Otolith-derived eye movements appear immediately after hatching, whereas the appearance and progressive amelioration of semicircular canal-evoked eye movements is delayed and dependent on the acquisition of sufficiently large semicircular canal diameters. Moreover, semicircular canal functionality is also required to tune the initially omnidirectional otolith-derived VOR. The tuning is due to a reinforcement of those vestibulo-ocular connections that are co-activated by semicircular canal and otolith inputs during natural head/body motion. This suggests that molecular mechanisms initially guide the basic ontogenetic wiring, whereas semicircular canal-dependent activity is required to establish the spatio-temporal specificity of the reflex. While a robust VOR is activated during passive head/body movements, locomotor efference copies provide the major source for compensatory eye movements during tail- and limb-based swimming of larval and adult frogs. The integration of active/passive motion-related signals for gaze stabilization occurs in central vestibular neurons that are arranged as segmentally iterated functional groups along rhombomere 1-8. However, at variance with the topographic maps of most other sensory systems, the sensory-motor transformation of motion-related signals occurs in segmentally specific neuronal groups defined by the extraocular motor output targets.

Amphibians are currently the most threatened and rapidly declining group of vertebrates and this has raised concerns about their potential sensitivity and exposure to plant protection products and other chemicals. Current environmental risk assessment procedures rely on surrogate species (e.g. fish and birds) to cover the risk to aquatic and terrestrial life stages of amphibians, respectively. Whilst a recent meta-analysis has shown that in most cases amphibian aquatic life stages are less sensitive to chemicals than fish, little research has been conducted on the comparative sensitivity of terrestrial amphibian life stages. Therefore, in this paper we address the questions "What is the relative sensitivity of terrestrial amphibian life stages to acute chemical oral exposure when compared with mammals and birds?" and "Are there correlations between oral toxicity data for amphibians and data for mammals or birds?" Identifying a relationship between these data may help to avoid additional vertebrate testing. Acute oral amphibian toxicity data collected from the scientific literature and ecotoxicological databases were compared with toxicity data for mammals and birds. Toxicity data for terrestrial amphibian life stages are generally sparse, as noted in previous reviews. Single-dose oral toxicity data for terrestrial amphibian life stages were available for 26 chemicals and these were positively correlated with LD50 values for mammals, while no correlation was found for birds. Further, the data suggest that oral toxicity to terrestrial amphibian life stages is similar to or lower than that for mammals and birds, with a few exceptions. Thus, mammals or birds are considered adequate toxicity surrogates for use in the assessment of the oral exposure route in amphibians. However, there is a need for further data on a wider range of chemicals to explore the wider applicability of the current analyses and recommendations., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Model organisms have long been important in biology and medicine due to their specific characteristics. Amphibians, especially Xenopus, play key roles in answering fundamental questions on developmental biology, regeneration, genetics, and toxicology due to their large and abundant eggs, as well as their versatile embryos, which can be readily manipulated and developed in vivo. Furthermore, amphibians have also proven to be of considerable benefit in human disease research due to their conserved cellular developmental and genomic organization. This review gives a brief introduction on the progress and limitations of these animal models in biology and human disease research, and discusses the potential and challenge of Microhyla fissipes as a new model organism.

The karst area of southwest China (KASC) is the largest piece of karst landscape on the earth and a global biodiversity hot-spot with high concentrations of endemic species. Although a number of nature reserves (NRs) have been established across the region, the representativeness of biodiversity of the NR system is still unknown. Based on comprehensive literature and field surveys, and intensive consultations with zoologists and wildlife managers, we compiled distributions of 1,204 terrestrial vertebrate species and 271 NRs in KASC. We found Jinxiu, Mengla, Hekou, and Jinghong have the richest amphibian species; Jinxiu has the highest species richness of reptiles; Jinghong, Menghai, and Mengla have the largest numbers of avian species; whereas, Mengla, Longzhou, and Ningming have the greatest mammalian diversity in the region. Gap analysis among NR system, species richness pattern, and five biogeographic indicators found insufficient representation of the NR system on territorial vertebrate diversity. The conservation effectiveness in Guizhou Province was much lower than that in Guangxi and Yunnan Provinces. Under-representation and over-representation simultaneously occurred in many of the ecoregions, elevation classes, vegetation types, landcover categories, and human disturbance intensity gradients. For conservation of terrestrial vertebrates in KASC, several suggestions were presented in this study.

Several reports suggested that rice seedling nursery-box application of some systemic insecticides (neonicotinoids and fipronil) is the cause of the decline in dragonfly species noted since the 1990s in Japan. We conducted paddy mesocosm experiments to investigate the effect of the systemic insecticides clothianidin, fipronil and chlorantraniliprole on rice paddy field biological communities. Concentrations of all insecticides in the paddy water were reduced to the limit of detection within 3 months after application. However, residuals of these insecticides in the paddy soil were detected throughout the experimental period. Plankton species were affected by clothianidin and chlorantraniliprole right after the applications, but they recovered after the concentrations decreased. On the other hand, the effects of fipronil treatment, especially on Odonata, were larger than those of any other treatment. The number of adult dragonflies completing eclosion was severely decreased in the fipronil treatment. These results suggest that the accumulation of these insecticides in paddy soil reduces biodiversity by eliminating dragonfly nymphs, which occupy a high trophic level in paddy fields.

The type specimens of amphibians and reptiles of the Museo de Historia Natural Alfredo Dugès, at the University of Guanajuato (MADUG) were reviewed following Smith & Necker's (1943) summary. Owing to this collection's eventful history and its historical importance as the oldest herpetological collection in Mexico, a review of its conservation status was needed. After many years, the collection has received proper recognition at the University of Guanajuato with a portion of the herpetological types considered "Precious Assets" of the university. We found 34 type specimens pertaining to 18 taxa; six are additional specimens to those previously reported; six herpetological types are missing, including the body of the type of Adelophis copei. All specimens are in good to reasonable condition except for the type of Rhinocheilus antonii, which has dried out completely. All specimens are illustrated to show their condition.

Extensive surveys at and around the type locality of Ichthyophis husaini Pillai & Ravichandran, 1999 (an 'unstriped' Ichthyophis) in North East India between 2008 and 2014 yielded tens of striped but no unstriped specimens of Ichthyophis. Contrary to the original description, we observed a lateral stripe on the holotype of I. husaini. The holotype of I. husaini is very similar to the holotype of the striped species I. garoensis Pillai & Ravichandran, 1999, which was collected from a locality only 20 km away from the type locality of I. husaini. The holotypes of these two species are also very similar to 12 newly collected specimens of striped Ichthyophis from this area. We conclude that the holotypes of I. husaini and I. garoensis represent but a single species, and we designate Ichthyophis husaini Pillai & Ravichandran, 1999 as a junior subjective synonym of Ichthyophis garoensis Pillai & Ravichandran, 1999. This taxonomic action and newly reported material extends the known geographic range of I. garoensis. It also reduces the number of Ichthyophis species of North East India to eight, and restricts unstriped members of the genus within this region to a single species (I. sikkimensis Taylor, 1960) that is known only from the far west of the region.

Forest buffers are a primary tool used to protect wetland-dependent wildlife. Though implemented widely, buffer efficacy is untested for most amphibian species. Consequently, it remains unclear whether buffers are sufficient for maintaining amphibian populations and if so, how wide buffers should be. We present evidence from a six-year, landscape-scale experiment testing the impacts of clearcutting, buffer width, and hydroperiod on body size and condition and biomass of breeding adults for two amphibian species at 11 vernal pools in the northeastern United States. We randomly assigned treatments (i.e., reference, 100m buffer, 30m buffer) across pools, clearcut to create buffers, and captured all spotted salamanders and wood frogs. Clearcuts strongly and negatively impacted size, condition, and biomass, but wider buffers mitigated effect magnitude and duration. Among recaptured individuals, for example, 30m-treatment salamanders were predicted to be about 9.5 mm shorter than, while 100m-treatment salamanders did not differ in length from, reference-treatment salamanders. Similarly, among recaptured frogs, mean length in the 30m treatment was predicted to decrease by about 1 mm/year, while in the 100m and reference treatments, length was time-invariant. Some, but not all, metrics recovered with time. For example, female new-captured and recaptured salamanders were predicted, respectively and on average, to weigh 4.5 and 7 g less in the 30m versus reference treatment right after the cut. While recaptured-female mass was predicted to recover by 9.5 years post-cut, new-captured-female mass did not recover. Hydroperiod was an important mediator: in the 100m treatment, cutting predominately affected pools that were stressed hydrologically. Overall, salamanders and female frogs were impacted more than male frogs. Our results highlight the importance of individualized metrics like body size, which can reveal sublethal effects and illuminate mechanisms by which habitat disturbance impacts wildlife populations. Individualized metrics thus provide critical insights that complement species occurrence and abundance-based population assessments.

Background: A model of thyroxine and metamorphosis of highland amphibians is frequently mentioned as an example of experiments on extremely diluted substances in discussions around 'homeopathy'., Methods: The model was scrutinized by reanalysing the results of the initial researcher A and a second researcher B as well as of 5 external researchers C between 1990 and 2013. Rana temporaria larvae were taken from an alpine highland biotope. The test solution was thyroxine 10(-30) (T30x), tetra-iodo-thyronine sodium pentahydrate diluted with pure water in 26 steps of 1:10, being agitated after each step. Analogously prepared water (W30x) was used for control. Tadpoles were observed from the 2-legged to the 4-legged stage. Experiments were performed in different years, at different times of season, and their duration could vary. Frequencies of 4-legged animals, effect sizes and areas under the curves (AUCs) were calculated and regression analyses were performed to investigate possible correlations between year, season, duration etc. Experiments were in line with animal protection guidelines., Results: The total set of data A + B + C as well as subsets A (initial researcher, N=286+293), B (second centre, 965 + 965) and C (5 external researchers, 690 + 690) showed an effect of extremely diluted agitated thyroxine reverse to that known of molecular thyroxin, i.e. test values were below control by 11.4% for A, 9.5% for B and 7.0% for C (p<0.001 for each of the subsets). The effect size (Cohen's d) was >0.8 (large) for both A and B and 0.74 (medium) for C., Conclusion: Although a perfect reproducibility was not obtained, this paradoxical phenomenon was generally consistent in different observations. Correlations were found between details of laboratory handling, as well as environment temperature, and the size of the results., (Copyright © 2015 The Faculty of Homeopathy. Published by Elsevier Ltd. All rights reserved.)

The order Caudata includes about 660 species and displays a variety of important developmental traits such as cleavage pattern and egg size. However, the cleavage process of tailed amphibians has never been analyzed within a phylogenetic framework. We use published data on the embryos of 36 species concerning the character of the third cleavage furrow (latitudinal, longitudinal or variable) and the magnitude of synchronous cleavage period (up to 3-4 synchronous cell divisions in the animal hemisphere or a considerably longer series of synchronous divisions followed by midblastula transition). Several species from basal caudate families Cryptobranchidae (Andrias davidianus and Cryptobranchus alleganiensis) and Hynobiidae (Onychodactylus japonicus) as well as several representatives from derived families Plethodontidae (Desmognathus fuscus and Ensatina eschscholtzii) and Proteidae (Necturus maculosus) are characterized by longitudinal furrows of the third cleavage and the loss of synchrony as early as the 8-cell stage. By contrast, many representatives of derived families Ambystomatidae and Salamandridae have latitudinal furrows of the third cleavage and extensive period of synchronous divisions. Our analysis of these ontogenetic characters mapped onto a phylogenetic tree shows that the cleavage pattern of large, yolky eggs with short series of synchronous divisions is an ancestral trait for the tailed amphibians, while the data on the orientation of third cleavage furrows seem to be ambiguous with respect to phylogeny. Nevertheless, the midblastula transition, which is characteristic of the model species Ambystoma mexicanum (Caudata) and Xenopus laevis (Anura), might have evolved convergently in these two amphibian orders.

The loss of genetic diversity, due to exposure to chemical contamination (genetic erosion), is a major threat to population viability. Genetic erosion is the loss of genetic variation: the loss of alleles determining the value of a specific trait or set of traits. Almost a third of the known amphibian species is considered to be endangered and a decrease of genetic variability can push them to the verge of extinction. This review indicates that loss of genetic variation due to chemical contamination has effects on: 1) fitness, 2) environmental plasticity, 3) co-tolerance mechanisms, 4) trade-off mechanisms, and 5) tolerance to pathogens in amphibian populations., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The decline of amphibian populations, particularly frogs, is often cited as an example in support of the claim that Earth is undergoing its sixth mass extinction event. Amphibians seem to be particularly sensitive to emerging diseases (e.g., fungal and viral pathogens), yet the diversity and geographic distribution of infectious agents are only starting to be investigated. Recent work has linked a previously undescribed protist with mass-mortality events in the United States, in which infected frog tadpoles have an abnormally enlarged yellowish liver filled with protist cells of a presumed parasite. Phylogenetic analyses revealed that this infectious agent was affiliated with the Perkinsea: a parasitic group within the alveolates exemplified by Perkinsus sp., a "marine" protist responsible for mass-mortality events in commercial shellfish populations. Using small subunit (SSU) ribosomal DNA (rDNA) sequencing, we developed a targeted PCR protocol for preferentially sampling a clade of the Perkinsea. We tested this protocol on freshwater environmental DNA, revealing a wide diversity of Perkinsea lineages in these environments. Then, we used the same protocol to test for Perkinsea-like lineages in livers of 182 tadpoles from multiple families of frogs. We identified a distinct Perkinsea clade, encompassing a low level of SSU rDNA variation different from the lineage previously associated with tadpole mass-mortality events. Members of this clade were present in 38 tadpoles sampled from 14 distinct genera/phylogroups, from five countries across three continents. These data provide, to our knowledge, the first evidence that Perkinsea-like protists infect tadpoles across a wide taxonomic range of frogs in tropical and temperate environments, including oceanic islands.

Size is a primary feature of biological systems that varies at many levels, from the organism to its constituent cells and subcellular structures. Amphibians populate some of the extremes in biological size and have provided insight into scaling mechanisms, upper and lower size limits, and their physiological significance. Body size variation is a widespread evolutionary tactic among amphibians, with miniaturization frequently correlating with direct development that occurs without a tadpole stage. The large genomes of salamanders lead to large cell sizes that necessitate developmental modification and morphological simplification. Amphibian extremes at the cellular level have provided insight into mechanisms that accommodate cell-size differences. Finally, how organelles scale to cell size between species and during development has been investigated at the molecular level, because subcellular scaling can be recapitulated using Xenopus in vitro systems., (Copyright © 2016 Cold Spring Harbor Laboratory Press; all rights reserved.)

The acquisition of sensory information by animals is central to species interactions. In aquatic environments, most taxa use chemical cues to assess predation risk and other key ecological factors. A number of laboratory studies suggest that anthropogenic pollutants can disrupt chemoreception, even when at low, non-toxic concentrations, but there are few tests of whether real-world variation in water quality affects chemoreception. Here we investigate whether chemosensory perception of predators by the gray treefrog, Hyla versicolor, depends on water quality. We evaluated the anti-predator response of anuran tadpoles housed in water collected from three sites that represent strong contrasts in the concentration and types of dissolved solids: de-chlorinated tap water, water from an impaired stream, and treated wastewater effluent. Behavioral assays were conducted in laboratory aquaria. Chemical cues associated with predation were generated by feeding tadpoles to dragonfly predators held in containers, and then transferring aliquots of water from dragonfly containers to experimental aquaria. Tadpoles housed in tap water responded to predator cues with an activity reduction of 49%. Tadpoles housed in stream water and wastewater effluent responded to predator cues by reducing activity by 29% and 24% respectively. The results of factorial ANOVA support the hypothesis that the response to predator cues depended on water type. These results show that alteration of the chemical environment can mediate chemical perception of predators in aquatic ecosystems. Because most aquatic species rely on chemoreception to gather information on the location of food and predators, any impairment of sensory perception likely has important ecological consequences.