Post-metamorphic Stages Research Articles

The impact of chemical pollution across major life transitions: a meta-analysis on oxidative stress in amphibians.

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.

Nor climate nor human impact factors: Chytrid infection shapes the skin bacterial communities of an endemic amphibian in a biodiversity hotspot.

The bacterial communities of the amphibian skin (i.e., the bacteriome) are critical to the host's innate immune system. However, it is unclear how different drivers can alter this function by modulating the bacteriome's structure. Our aim was to assess the extent to which different host attributes and extrinsic factors influence the structure of the bacterial communities of the skin. Skin bacterial diversity was examined in 148 individuals of the four-eyed frog (Pleurodema thaul) from 16 localities spanning almost 1800 km in latitude. The richness and beta diversity of bacterial families and the richness and abundance of Bd-inhibitory bacterial genera were used to describe their structure. Predictors associated with the host (developmental stage, genetic lineage, individual Batrachochytrium dendrobatidis [Bd] infection status) and the landscape (current climate, degree of anthropogenic disturbance) were used in the statistical modeling in an information theoretical approach. Bd infection and host developmental stage were the only predictors affecting bacteriome richness, with Bd+ individuals and postmetamorphic stages (adults and juveniles) having higher richness than Bd- ones and tadpoles. High diversity in Bd+ individuals is not driven by bacterial genera with known anti-Bd properties. Beta diversity was not affected by Bd infection and was mainly a consequence of bacterial family turnover rather than nestedness. Finally, for those bacterial genera known to have inhibitory effects on chytrid, Bd+ individuals had a slightly higher diversity than Bd- ones. Our study confirms an association between Bd infection and the host developmental stage with the bacterial communities of the skin of P. thaul. Unexpectedly, macroclimate and human impact factors do not seem to play a role in shaping the amphibian skin microbiome. Our study exemplifies that focusing on a single host-parasite system over a large geographic scale can provide essential insights into the factors driving host-parasite-bacteriome interactions.

Ontogeny drives shifts in skin bacterial communities in facultatively paedomorphic salamanders.

Microbiomes are major determinants of host growth, development and survival. In amphibians, host-associated bacteria in the skin can inhibit pathogen infection, but many processes can influence the structure and composition of the community. Here we quantified the shifts in skin-associated bacteria across developmental stages in the striped newt (Notophthalmus perstriatus), a threatened salamander species with a complex life history and vulnerable to infection by the amphibian chytrid fungus Batrachochytrium dendrobatidis and ranavirus. Our analyses show that pre-metamorphic larval and paedomorphic stages share similar bacterial compositions, and that the changes in the microbiome coincided with physiological restructuring during metamorphosis. Newts undergoing metamorphosis exhibited microbiome compositions that were intermediate between paedomorphic and post-metamorphic stages, further supporting the idea that metamorphosis is a major driver of host-associated microbes in amphibians. We did not find support for infection-related disruption of the microbiome, though infection replicates were small for each respective life stage.

Genesis and Evolution of the Yolindi Cu-Fe Skarn Deposit in the Biga Peninsula (NW Turkey): Insights from Genetic Relationships with Calc-Alkaline Magmatic Activity

The current work investigates the impact of magmatic fluids and metasomatic processes on the Yolindi Cu-Fe skarn deposit in the Biga Peninsula, Turkey. It traces the stages of skarn evolution, from prograde to retrograde alterations, and investigates findings within a broader geological, mineralogical, and geochemical framework. Additionally, it assesses the evolutionary history of the Yolindi deposit in relation to calc-alkaline magmatic activity in an island-arc environment and compares its mineral compositions and genesis with other global and regional Cu-Fe skarn deposits. The Yolindi Cu-Fe skarn deposit in the Biga Peninsula was formed by the intrusion of Şaroluk quartz monzonite pluton into Upper Paleozoic Torasan Formation rocks such as phyllite, schists, hornfels, marble, and serpentinites. During skarnification, reactions between the magmatic fluids from the Şaroluk quartz monzonite pluton and the Torasan Formation produced skarn minerals associated with metals such as Fe and Cu. Initially, these reactions formed prograde skarn minerals such as augite-rich pyroxenes and andradite garnets with magnetite and pyrite. As the system cooled, these initial minerals underwent retrograde alteration, leading to the formation of minerals such as epidote, actinolite, and chlorite, as well as other copper and iron minerals including chalcopyrite, bornite, secondary magnetite, and specular hematite. Therefore, four main stages influenced the formation of the Yolindi Cu-Fe deposit: metamorphic bimetasomatic, prograde metasomatic, and retrograde metasomatic stages. Later, oxidation and weathering resulted in supergene minerals such as cerussite, malachite, and goethite, which serve as examples of the post-metamorphic stage. The mineralogical shifts, such as the andradite–grossular transition, reflect changing hydrothermal fluid compositions and characteristics due to the addition of meteoric fluids. Importantly, the formation of magnetite after garnet and clinopyroxene during the retrograde stage is evidenced by magnetite crystals within garnet. The mineral associations of the Yolindi Cu-Fe skarn deposit align with the global skarn deposits and specific Turkish skarns (e.g., Ayazmant Fe-Cu and Evciler Cu-Au skarn deposits). The Yolindi Cu-Fe skarn deposit, in association with ore-bearing solutions having magmatic origins, developed in an island-arc setting.

Post-thaw quality assessment of testicular fragments as a source of spermatogonial cells for surrogate production in the flatfish Solea senegalensis.

Cryopreservation of germ cells would facilitate the availability of cells at any time allowing the selection of donors and maintaining quality control for further applications such as transplantation and germline recovery. In the present study, we analyzed the efficiency of four cryopreservation protocols applied either to isolated cell suspensions or to testes fragments from Senegalese sole. In testes fragments, the quality of cryopreserved germ cells was analyzed in vitro in terms of cell recovery, integrity and viability, DNA integrity (fragmentation and apoptosis), and lipid peroxidation (malondialdehyde levels). Transplantation of cryopreserved germ cells was performed to check the capacity of cells to in vivo incorporate into the gonadal primordium of Senegalese sole early larval stages (6 days after hatching (dah), pelagic live), during metamorphosis (10 dah) and at post-metamorphic stages (16 dah and 20 dah, benthonic life). Protocols incorporating dimethyl sulfoxide (DMSO) as a cryoprotectant showed higher number of recovered spermatogonia, especially in samples cryopreserved with L-15 + DMSO (0.39 ± 0.18 × 106 cells). Lipid peroxidation and DNA fragmentation were also significantly lower in this treatment compared with other treatments. An important increase in oxidation (MDA levels) was detected in samples containing glycerol as a cryoprotectant, reflected also in terms of DNA damage. Transplantation of L-15 + DMSO cryopreserved germ cells into larvae during early metamorphosis (10 dah, 5.2 mm) showed higher incorporation of cells (27.30 ± 5.27%) than other larval stages (lower than 11%). Cryopreservation of germ cells using testes fragments frozen with L-15 + DMSO was demonstrated to be a useful technique to store Senegalese sole germline.

Genome-wide chromatin accessibility and gene expression profiling during flatfish metamorphosis

Metamorphosis is a widely studied post-embryonic process in which many tissues undergo dramatic modifications to adapt to the new adult lifestyle. Flatfishes represent a good example of metamorphosis in teleost fishes. During metamorphosis of flatfish, organ regression and neoformation occur, with one of the most notable changes being the migration of one of the eyes to the other side of the body. In order to create a useful and reliable tool to advance the molecular study of metamorphosis in flatfish, we generated a chromatin accessible atlas as well as gene expression profile during four developmental stages ranging from a phylotypic to a post-metamorphic stage. We identified 29,019 differentially accessible chromatin regions and 3,253 differentially expressed genes. We found stage-specific regulatory regions and gene expression profiles, supporting the quality of the results. Our work provides strongly reproducible data for further studies to elucidate the regulatory elements that ensure successful metamorphosis in flatfish species.

Histological changes of the skin during postembryonic development of the crested newt Triturus ivanbureschi (Urodela, Salamandridae)

BackgroundAmphibian skin has been studied for many decades, especially the metamorphic changes in the skin of frogs. Less attention has been paid to salamander skin. Here, we describe changes in the skin structure during postembryonic development in a salamandrid species, the Balkan crested newt Triturus ivanbureschi. MethodUsing traditional histological techniques we examined the skin in the trunk region of three premetamorphic larval stages (hatchling, mid larval and late larval) and two postmetamorphic stages (juvenile, just after metamorphosis, and adult). ResultsIn larval stages, skin consists only of the epidermis, which gradually develops from the single epithelial cell layer in hatchlings, to a stratified epidermis with gland nests and characteristic Leydig cells at the late larval stage. During metamorphosis, Leydig cells disappear, and the dermal layer develops. In postmetamorphic stages, skin is differentiated on stratified epidermis and the dermis with well-developed glands. Three types of glands were observed in the skin of the postmetamorphic stages: mucous, granular and mixed. Gland composition appears to be stage- and sex-specific, with juveniles and adult female being more similar to each other. In juveniles and adult female, there are a similar proportion of glands in both dorsal and ventral skin, whereas in adult male granular glands dominated the dorsal skin, while mixed glands dominated the ventral skin. ConclusionOur results provide a baseline for future comparative research of skin anatomy in salamanders.

Influence of pyrethroid and neonicotinoid insecticides on post-metamorphic amphibians (literature review)

Agricultural activity in the global world is accompanied by the use of a significant number of synthetic insecticides for the control of insect pests. Pyrethroid and neonicotinoid insecticides are among the widely used insecticides in many countries for the control of crop pests. They are a generation of synthetic insecticides that have replaced the more environmentally stable organophosphorus and organochlorine compounds. Pyrethroid and neonicotinoid insecticides were thought to have low toxicity to vertebrates, leading to their widespread use and increased production. However, many studies have emerged in recent decades that have shown that, under certain conditions, these substances can cause significant damage to the internal systems of amphibians. Recently, special studies have also revealed the toxic effects of pyrethroids and neonicotinoids on the post-metamorphic stages of amphibians, which had previously been ignored. It has also been noted that abnormalities in gastrointestinal tract functions occur, leading to abnormalities in the digestive system. Pyrethroid and neonicotinoid insecticides have been shown to affect the biochemical and histological parameters of amphibians. The possible genotoxicity of these insecticides resulted in producing erythrocytes with abnormal nuclei and an increased number of micronuclei in amphibian cells. Meanwhile, changes in the activity of antioxidant enzymes and increases in lipid peroxidation products could be used as biomarkers of oxidative stress in amphibians under the influence of pyrethroid and neonicotinoid insecticides. The available literature also indicates that these insecticides appear to affect the nervous system of amphibians and induce changes in their behaviour. At the same time, our data suggest that it is neuromolecular biomarkers that can be practised to determine the toxic effects of insecticides on non-target species. Such biomarkers can be used in the context of the low-dose influence of insecticides, which however requires additional research on amphibians.

Early exposure to UV radiation causes telomere shortening and poorer condition later in life.

ABSTRACTDetermining the contribution of elevated ultraviolet-B radiation (UVBR; 280–315 nm) to amphibian population declines is being hindered by a lack of knowledge about how different acute UVBR exposure regimes during early life-history stages might affect post-metamorphic stages via long-term carryover effects. We acutely exposed tadpoles of the Australian green tree frog (Litoria caerulea) to a combination of different UVBR irradiances and doses in a multi-factorial laboratory experiment, and then reared them to metamorphosis in the absence of UVBR to assess carryover effects in subsequent juvenile frogs. Dose and irradiance of acute UVBR exposure influenced carryover effects into metamorphosis in somewhat opposing manners. Higher doses of UVBR exposure in larvae yielded improved rates of metamorphosis. However, exposure at a high irradiance resulted in frogs metamorphosing smaller in size and in poorer condition than frogs exposed to low and medium irradiance UVBR as larvae. We also demonstrate some of the first empirical evidence of UVBR-induced telomere shortening in vivo, which is one possible mechanism for life-history trade-offs impacting condition post-metamorphosis. These findings contribute to our understanding of how acute UVBR exposure regimes in early life affect later life-history stages, which has implications for how this stressor may shape population dynamics.

Carryover effects of chronic exposure to ammonium during the larval stage on post-metamorphic frogs.

Water contamination poses an important challenge to aquatic fauna, including well-documented effects on amphibian larvae. However, little is known about how contamination during the larval stages may affect post-metamorphic phases, or whether resistance may have evolved in some populations. In this work, we tested the hypothesis that chronic exposure to ammonium (a common contaminant in agroecosystems with confirmed effects on anuran tadpoles) during the larval stage of Pelophylax perezi frogs would affect growth and locomotor performance of metamorph, juvenile, subadult and adult stages. We also predicted that the effects of ammonium would be milder in offspring originated from parental agroecosystem frogs than those originating from forests. We compared tadpoles from both habitats either reared in untreated water or chronically exposed to ammonium. We found that exposure to ammonium during the larval stage inflicted effects on morphology (different measures of body size) and swimming speed after metamorphosis until adulthood. However, these effects were not always consistent through post-metamorphic stages and the effects differed as a function of treatment and habitat. In adults, body size and condition were greater in non-ammonium and ammonium exposed individuals, respectively. These differences were not detectable in metamorphs, for which only ammonium-exposed individuals from agroecosystem showed reduced body size in intermediate post-metamorphic stages. In turn, treatment reduced jumping distance only in agroecosystem adults, subadults and juveniles, which was opposite to the trend observed just after metamorphosis. These changes of patterns throughout the ontogeny of P. perezi could be due to processes such as compensatory growth, delayed energy costs derived from it, or early sexual differences that could be present even before they can be accounted for. In summary, this study suggests that exposure to ammonium during larval stages can result in diverse biological and long-term outcomes in later life stages.

Vertical distribution patterns of early stages of mesopelagic fishes along 110 °E, south-east Indian Ocean

This study examines vertical distribution of the early stages of mesopelagic fishes in the south-eastern Indian Ocean and oceanographic factors that may affect it. This investigation along the 110 °E transect formed part of the second International Indian Ocean Expedition (IIOE-2). Depth-stratified samples to 500 m depth of early stages of mesopelagic fishes were obtained by means a multinet EZ-1 m2. This net captured larvae and transforming stages, but juveniles and adults were also caught, particularly at night. Diel vertical patterns of the different species and stages were analysed relative to the oceanographic conditions encountered at the 20 stations along the transect. This paper provides the ranges of vertical occurrence and weighted mean depth for larvae (135 taxa), and post-metamorphic stages (75 taxa) of fishes caught during the survey. Despite the varying vertical profiles of oceanographic parameters encountered, there were consistent patterns within species and families along the transect, with broader vertical distribution of larvae at the southern stations, where the upper mixed layer was deeper. During both day and night, larval stages of all the species were concentrated in the epipelagic zone, with some differences in terms of weighted mean depth (WMD) between taxa, but with no significant differences in vertical location between day and night. Transforming and juvenile-adult stages, however, presented a wider vertical distribution range than larval stages, and showed important differences among families and species. For example, Myctophidae and Phosichthyidae species presented the highest concentrations in the upper 50–100 m at night, and a much deeper distribution during the day, thereby indicating night vertical migration behaviour of transforming and juvenile-adult stages. By contrast, during these stages the Gonostomatidae Cyclothone spp. and the Sternoptychidae Argyropelecus spp. occurred deeper than the epipelagic layer during both day and night. Although these vertical patterns concur with results from other oceans, the WMD along 110 °E was slightly deeper than that observed in other areas, such as the eastern Atlantic, possibly related to water transparency in this oligotrophic part of the world's oceans.

Internal controls for quantitative RT-PCR analysis of gene expression in response to ocean acidification in edible oysters

The increase of CO2 by anthropogenic activities leads to a decrease of pH in the ocean surface due to ocean acidification (OA) process. Generally, OA not only reduces the rate of calcification in marine environments but also affects various physiological activities, especially in calcifiers, including edible oysters. Quantitative real-time PCR (qRT-PCR) is often used to detect gene expression in response to OA, which relies on the stability of internal control. However, the appropriate internal controls for OA experiments remain scarce especially in the marine calcifiers. Hence, this study developed internal controls for qRT-PCR assays using the Hong Kong oyster (Crassostrea hongkongensis) as a model to reveal gene expression profile during development under OA. In this study, 17 housekeeping genes were selected as the possible candidate of the internal controls. After a comprehensive interpretation from the multiple algorithms and software, GAPDH paired with RL23 is recommended for the normalization for planktonic larvae and benthic juveniles, but beyond that, TUBB and EF2 are recommended for post-metamorphic stage. Moreover, GAPDH and EF2 were suitable for various pH treatments, and TUBB, RL35 and RL23 could be the alternatives for OA experiments. These results are instrumental for the selection of internal control in Crassostrea hongkongensis during the development, and shed light on other molecular OA experiments in marine invertebrates for reference.

Carry-Over Effects of Desiccation Stress on the Oxidative Status of Fasting Anuran Juveniles.

Amphibians are sensitive to deteriorating environmental conditions, especially during transition to a terrestrial environment which is full of uncertainties. Harsh conditions, such as desiccation during earlier stages, affect different larval traits with possible carry-over effects on juvenile and adult life histories. The first consequences of the effects can be seen in juveniles in the challenges to find food and the ability to survive without it in a terrestrial habitat. Body size and the internal energy reserves acquired during the larval phase play an important role in this period. Herein, we tested how different water regimes (low water availability, desiccation and constant high-water availability) during larval development reflect on the oxidative status and ability of yellow belly toad (Bombina variegata) juveniles to endure short-term fasting. The desiccation regime significantly reduced the body size of metamorphs. The same was observed after 2 weeks of fasting, while the feeding treatment reduced differences mostly in the body mass of individuals from different water regimes. This was the result of a greater gain in mass in juveniles pre-exposed to desiccation. Pre-exposure to desiccation also modified the parameters of the antioxidant system (AOS) under feeding conditions, leading to higher values of superoxide dismutase, glutathione reductase and glutathione S-transferase, glutathione and sulfhydryl group concentrations, and lower glutathione peroxidase in comparison to juveniles reared under constant water. The increase in the AOS of juveniles can be considered as a physiological carry-over effect of desiccation, probably as the result of compensatory growth and/or earlier exposure to chronic stress. However, water levels during larval development did not exert significant effects on the oxidative status of juveniles subjected to food unavailability. Fasting juveniles, both control and desiccated, were exposed to oxidative stress, significantly higher lipid peroxide concentrations, lower superoxide dismutase, glutathione peroxidase, glutathione S-transferase, glutathione and sulfhydryl group values in comparison to feeding individuals. The lack of food in juvenile anurans activated the AOS response in the same manner, regardless of body size and stress pre-exposure, suggesting that the generally accepted hypothesis about the influence of metamorphic body size on the fitness of the postmetamorphic stage should be tested further.

Changes in marine turbot (Scophthalmus maximus) epidermis and skin mucus composition during development from bilateral larvae to juvenile flat fish.

Alike other flat fish, marine turbot has the particularity that changes from larvae with bilateral symmetry to adult with asymmetry, in terms of the position of the eyes. As expected, the skin configuration of this species is also affected by the development and transformation suffered by fish during metamorphosis. In this context, changes in the epidermis of marine turbot were studied using conventional staining and histochemical techniques using six lectins (UEA-I, PNA, RCA-I, WGA, Con A and SBA). During development from larvae to juvenile (3-300 days post-hatching), the epidermis increased in both thickness and the number of cell layers. In fact, the simple cuboidal epithelium observed in larvae at day 3 already became stratified at days 10-12, which sequentially increase in thickness with fish development. Turbot epidermis is composed basically of four cell types: epithelial and mucous or secretory cells that are present through the development, and pigmented cells and a type that the authors described as club-like cells that appear during and post-metamorphosis. The Alcian blue-periodic acid Schiff (AB-PAS) histochemical method revealed the presence of neutral glycoconjugates in mucous and club-like cells at post-metamorphic stages of fish. Accordingly, lectin analysis showed mucous cells containing glycoproteins rich in fucose (UEA-I labelling) and glycoconjugates rich in the sequence galactose-N-acetyl galactosamine (PNA and RCA-I labelling) when this cell type appears. Interestingly, melanophores were observed in the dorsal epidermis of post-metamorphic juveniles. This type of cell contains a black-to-brown pigment that provides the skin the typical colour of this fish species. Changes in mucous coat composition were observed during fish development, which was attributed to different roles of the glycoconjugates.

A Small Proportion of Breeders Drive American Bullfrog Invasion of the Yellowstone River Floodplain, Montana

The American bullfrog (Lithobates catesbeianus) is a non-native invader of aquatic habitats across the northwestern United States. It recently invaded the Yellowstone River, Montana, and has spread to over 140 km of floodplain habitat. We analyzed seven microsatellites in 528 tadpoles sampled across nearly the entire Yellowstone River invasion (about 140 river km) to characterize invasion genetics, compare our results with those of a recent mtDNA study (Kamath et al. 2016), and to inform control efforts. Microsatellite variation supports the mtDNA-based hypothesis of at least two independent introductions to the floodplain from genetically divergent populations in the midwestern United States, followed by massive range expansion. One introduction is associated with the upstream extent of the invasion near Park City, Montana and the other more broadly with downstream populations. All sites were characterized by small effective numbers of breeders (Nb; harmonic mean = 9.97), and therefore, a small proportion of highly successful adults may drive the invasion by producing large families. Microsatellites and mtDNA produced discordant estimates of genetic admixture between the upstream and downstream invasions, which may reflect small effective population size. Although we observed isolation by distance using both types of markers, microsatellites appear to reflect population structure resulting from secondary contact between the two introductions, as opposed to structure resulting from equilibrium between gene flow and genetic drift. Most sites showed evidence for genetic bottlenecks, which supports the recent history of invasion. Small Nb paired with known high localized extinction rates following colonization suggests focused removal of post metamorphic life stages at sites less likely to go extinct on their own could help limit invasion by bullfrogs.

Patrón de expresión tisular de los genes elovl4 en Sparus aurata y Solea senegalensis: de larva a adulto

Very long-chain (˃ C24) fatty acids (VLC-FA) play critical roles during early development of vertebrates, since these compounds are accumulated in the rapidly forming neural tissues, ensuring their normal function. The functionality of VLC-FA has aroused scientific interest, focusing on the study of elongases protein 4 of very long chain fatty acid (Elovl4), which are responsible for their biosynthesis from shorter fatty acids (precursors). For a better understanding of the metabolism and the potential tissue-specific requirements of VLC-FA in marine teleosts, the present study aimed to determine the tissue-expression pattern of the genes that encode for Elovl4 isoforms, i.e. elovl4a and elovl4b, in different windows of development (larval and adult stages) of Gilthead seabream Sparus aurata and Senegalese sole Solea senegalensis. The results indicated that in S. aurata larvae, elovl4a is widely expressed in the head, while elovl4b is strongly focused in the eyes. Interestingly, in S. senegalensis larvae an opposite tissue-expression pattern was observed for both elovl4 isoforms. In adults of both fish, although elovl4 transcripts were detected in most tissues analyzed, elovl4a and elovl4b genes were strongly expressed in brain and eyes, respectively. Importantly, the differential tissue-expression pattern of both elovl4 isoforms associated to the pre- and post-metamorphic stage of S. senegalensis could be indicative of the VLC-FA particular needs linked to neural tissues functionality in each development stage. These findings can contribute to a better understanding of the species-specific VLC-FA metabolism in marine teleosts.

Review of post-metamorphic frog-eat-frog predation, with a description of a new cases of anurophagy

Predatory interactions among anurans (Lissamphibia; Order: Anura) within food webs are important to assessing mortality threats in a conservation context. Anurans are a significant prey item for other anurans in post-metamorphic stage; however, to date, there are few studies addressing the distribution of frog-eat-frog predatory behavior across anuran taxa. Therefore, in this work, we review frog-eat-frog predation (hereafter, post-metamorphic anurophagy) in post-metamorphic individuals. Moreover, we describe the first case of anurophagy by the Colombian endemic frog Craugastor metriosistus preying on a juvenile of the Truando Toad Rhaebo haematiticus and on a juvenile of unidentified frog. In the literature, we found 161 records of anurophagy among 59 anuran predator species belonging to 29 genera and 16 families. Further, 134 species from 62 genera and 22 families were recorded as anuran prey. Anurophagy is mainly observed in large-bodied species belonging to families with generalist behaviors, such as Ceratophryidae, Leptodactylidae, and Ranidae. We also descibe the first record of anurophagy in the family Craugastoridae and suggest that this intra-Order predatory behavior might not be widespread across anuran taxa.

Robustness of life histories to environmental variability in complex versus simple life cycles

Most animal species have a complex life cycle (CLC) with metamorphosis. It is thus of interest to examine possible benefits of such life histories. The prevailing view is that CLC represents an adaptation for genetic decoupling of juvenile and adult traits, thereby allowing life stages to respond independently to different selective forces. Here I propose an additional potential advantage of CLCs that is, decreased variance in population growth rate due to habitat separation of life stages. Habitat separation of pre- and post-metamorphic stages means that the stages will experience different regimes of environmental variability. This is in contrast to species with simple life cycles (SLC) whose life stages often occupy one and the same habitat. The correlation in the fluctuations of the vital rates of life stages is therefore likely to be weaker in complex than in simple life cycles. By a theoretical framework using an analytical approach, I have (1) derived the relative advantage, in terms of long-run growth rate, of CLC over SLC phenotypes for a broad spectrum of life histories, and (2) explored which life histories that benefit most by a CLC, that is avoid correlation in vital rates between life stages. The direction and magnitude of gain depended on life history type and fluctuating vital rate. One implication of our study is that species with CLCs should, on average, be more robust to increased environmental variability caused by global warming than species with SLCs.

A comparison of the ultrastructure of the cornea of the pre- and post-metamorphic axolotl (Ambystoma mexicanum, Amphibia)

The corneal ultrastructure of the pre- and post-metamorphic stages of the neotenic axolotl Ambystoma mexicanum is examined using light microscopy and both scanning and transmission electron microscopy to reveal whether there are any morphological changes associated with a switch in lifestyle. Although the complement of corneal layers remains the same, there are significant quantitative changes in corneal, epithelial and stromal thickness, epithelial and endothelial cell size and density, and the thickness of Bowman's layer and Desçemet's membrane. Microholes in the epithelium and vertical sutures within the stroma are predominant features in the pre-metamorphic stage but are rarely seen in the post-metamorphic stage. There are also significant quantitative centro-peripheral differences in the thickness of the whole cornea, primarily due to differences in the thickness of the stroma in both metamorphic stages. These changes may reflect the physiological demands on the cornea as it switches from a purely aquatic to an amphibious lifestyle, which includes venturing onto land.

The first report on circulating microRNAs at Pre- and Post-metamorphic stages of axolotls.

MicroRNAs (miRNAs) are endogenously coded small RNAs, implicated in post-transcriptional gene regulation by targeting messenger RNAs (mRNAs). Circulating miRNAs are cell-free molecules, found in body fluids, such as blood and saliva, and emerged recently as potential diagnostic biomarkers. Functions of circulating miRNAs and their roles in target tissues have been extensively investigated in mammals, and the reports on circulating miRNAs in non-mammalian clades are largely missing. Salamanders display remarkable regenerative potential, and the Mexican axolotl (Ambystoma mexicanum), a critically endangered aquatic salamander, has emerged as a powerful model organism in regeneration and developmental studies. This study aimed to explore the circulating miRNA signature in axolotl blood plasma. Small RNA sequencing on plasma samples revealed 16 differentially expressed (DE) circulating miRNAs between neotenic and metamorphic stages out of identified 164 conserved miRNAs. Bioinformatics predictions provided functional annotation of detected miRNAs for both stages and enrichment of DE miRNAs in cancer-related and developmental pathways was notable. Comparison with previous reports on axolotl miRNAs unraveled common and unique members of the axolotl circulating miRNome. Overall, this work provides novel insights into non-mammalian aspects of circulating miRNA biology and expands the multi-omics toolkit for this versatile model organism.