Impact of Plant Cover on Fitness and Behavioural Traits of Captive Red-Eyed Tree Frogs (Agalychnis callidryas)

Despite the importance of ex situ conservation programmes as highlighted in the Amphibian Conservation Action Plan, there are few empirical studies that examine the influence of captive conditions on the fitness of amphibians, even for basic components of enclosure design such as cover provision. Maintaining the fitness of captive amphibian populations is essential to the success of ex situ conservation projects. Here we examined the impact of plant cover on measures of fitness and behaviour in captive red-eyed tree frogs (Agalychnis callidryas). We found significant effects of plant provision on body size, growth rates and cutaneous bacterial communities that together demonstrate a compelling fitness benefit from cover provision. We also demonstrate a strong behavioural preference for planted rather than non-planted areas. We also assessed the impact of plant provision on the abiotic environment in the enclosure as a potential driver of these behavioural and fitness effects. Together this data provides valuable information regarding enclosure design for a non-model amphibian species and has implications for amphibian populations maintained in captivity for conservation breeding programmes and research.     

A comparison of understanding of the amphibian crisis by zoo visitors across three countries

Despite the global declines in the rate of amphibians, evaluation of public understanding of the crisis has not yet been carried out. We surveyed visitors (n = 1,293) at 15 zoos in Brazil, New Zealand, and the United Kingdom, using a certainty‐based assessment method to compare visitor knowledge of the global amphibian crisis. We further analyzed zoo educational material about amphibians to explore its potential to raise awareness through amphibian‐focused environmental education. Visitors in the three countries had relatively little understanding of amphibians and the global amphibian crisis. When the degree of confidence in answering the questions (high, medium, and low) is accounted for, correct answers varied between 28% and 39%. This compared to scores of between 58% and 73% when the degree of confidence in responding was not accounted for. However, specific areas of knowledge (e.g., biology, conservation, biogeography, and conceptual ideas) varied significantly across the countries. Visitors had a weaker grasp of biogeographical and conservation issues than general amphibian biology. Zoo visitors in Brazil knew less about amphibian conservation than those in New Zealand or the United Kingdom. There was less amphibian‐focused content in educational materials in zoos in Brazil than there was in the United Kingdom. Improving information about the global amphibian crisis may increase support for future conservation actions. Outreach education is one of the most important approaches in any strategic planning for conservation of species. Amphibian‐focused environmental education at institutions such as zoos and aquaria can be a crucial intervention to support amphibian conservation worldwide.     

Batrachochytrium dendrobatidis infection of amphibians in the Doñana National Park, Spain

Amphibian chytridiomycosis, caused by infection with the non-hyphal, zoosporic chytrid fungus Batrachochytrium dendrobatidis (Bd), is an emerging infectious disease recognised as a cause of recent amphibian population declines and extinctions worldwide. The Do?ana National Park (DNP) is located in southwestern Spain, a country with widespread Bd infection. This protected area has a great diversity of aquatic habitats that constitute important breeding habitats for 11 native amphibian species. We sampled 625 amphibians in December 2007 and February to March 2008, months that correspond to the early and intermediate breeding seasons for amphibians, respectively. We found 7 of 9 sampled species to be infected with Bd and found differences in prevalence between sampling periods. Although some amphibians tested had higher intensities of infection than others, all animals sampled were apparently healthy and, so far, there has been no evidence of either unusually high rates of mortality or amphibian population declines in the DNP.     

Hurricane storm surge and amphibian communities in coastal wetlands of northwestern Florida

Isolated wetlands in the Southeastern United States are dynamic habitats subject to fluctuating environmental conditions. Wetlands located near marine environments are subject to alterations in water chemistry due to storm surge during hurricanes. The objective of our study was to evaluate the effect of storm surge overwash on wetland amphibian communities. Thirty-two wetlands in northwestern Florida were sampled over a 45-month period to assess amphibian species richness and water chemistry. During this study, seven wetlands were overwashed by storm surge from Hurricane Dennis which made landfall 10 July 2005 in the Florida panhandle. This event allowed us to evaluate the effect of storm surge overwash on water chemistry and amphibian communities of the wetlands. Specific conductance across all wetlands was low pre-storm (<100 μS/cm), but increased post-storm at the overwashed wetlands ([`(x)] \bar{x}  = 7,613 μS/cm). Increased specific conductance was strongly correlated with increases in chloride concentrations. Amphibian species richness showed no correlation with specific conductance. One month post-storm we observed slightly fewer species in overwashed compared with non-overwashed wetlands, but this trend did not continue in 2006. More species were detected across all wetlands pre-storm, but there was no difference between overwashed and non-overwashed wetlands when considering all amphibian species or adult anurans and larval anurans separately. Amphibian species richness did not appear to be correlated with pH or presence of fish although the amphibian community composition differed between wetlands with and without fish. Our results suggest that amphibian communities in wetlands in the southeastern United States adjacent to marine habitats are resistant to the effects of storm surge overwash.     

Calling phenology and detectability of a threatened amphibian (Litoria olongburensis) in ephemeral wetlands varies along a latitudinal cline: Implications for management

Amphibian reproductive biology and activity are highly sensitive to environmental cues. For amphibian conservation in the face of climate change, a clear priority is to investigate impacts of variable weather on breeding activity and detectability. This study assessed variation in the phenology, intensity and detectability of calling for a threatened temporary wetland breeding species, Litoria olongburensis. Acoustic and visual surveys were conducted monthly from August 2009 to July 2011 over the species latitudinal range in eastern Australia. Calling phenology of L. olongburensis varied spatially and temporally within and among latitudinal clusters. Detectability of frogs also varied among months and transects, and also with the type of survey conducted (visual vs. acoustic). Air temperature was a strong predictor for the onset of the calling season (early spring); however, peaks in calling abundances were primarily driven by monthly accumulated rainfall. The duration of the calling season varied among latitudinal clusters, becoming narrower moving south, reflecting clinal variation in weather patterns. This study presents a framework for amphibian species management using a combination of survey techniques and the climatic conditions influencing detectability. Applied along the latitudinal gradient, it provides data for predicting the likely impacts of climate change on amphibian activity and reproduction.     

Heterogeneous Vesicles in Mucous Epithelial Cells of Posterior Esophagus of Chinese Giant Salamander (Andrias Davidianus)

The Chinese giant salamander belongs to an old lineage of salamanders and endangered species. Many studies of breeding and disease regarding this amphibian had been implemented. However, the studies on the ultrastructure of this amphibian are rare. In this work, we provide a histological and ultra-structural investigation on posterior esophagus of Chinese giant salamander. The sections of amphibian esophagus were stained by hematoxylin & eosin (H&E). Moreover, the esophageal epithelium was observed by transmission electron microscopy (TEM). The results showed that esophageal epithelium was a single layer epithelium, which consisted of mucous cells and columnar cells. The esophageal glands were present in submucosa. The columnar cells were ciliated. According to the diverging ultrastructure of mucous vesicles, three types of mucous cells could be identified in the esophageal mucosa: i) electron-lucent vesicles mucous cell (ELV-MC); ii) electron-dense vesicles mucous cell (EDV-MC); and iii) mixed vesicles mucous cell (MV-MC).Key words: Amphibian, esophagus, ultrastructure, mucous cells, heterogeneous vesicles     

Amphibian skin fungal communities vary across host species and do not correlate with infection by a pathogenic fungus

Amphibian population declines caused by the fungus Batrachochytrium dendrobatidis (Bd) have prompted studies on the bacterial community that resides on amphibian skin. However, studies addressing the fungal portion of these symbiont communities have lagged behind. Using ITS1 amplicon sequencing, we examined the fungal portion of the skin microbiome of temperate and tropical amphibian species currently coexisting with Bd in nature. We assessed cooccurrence patterns between bacterial and fungal OTUs using a subset of samples for which bacterial 16S rRNA gene amplicon data were also available. We determined that fungal communities were dominated by members of the phyla Ascomycota and Basidiomycota, and also by Chytridiomycota in the most aquatic amphibian species. Alpha diversity of the fungal communities differed across host species, and fungal community structure differed across species and regions. However, we did not find a correlation between fungal diversity/community structure and Bd infection, though we did identify significant correlations between Bd and specific OTUs. Moreover, positive bacterial–fungal cooccurrences suggest that positive interactions between these organisms occur in the skin microbiome. Understanding the ecology of amphibian skin fungi, and their interactions with bacteria will complement our knowledge of the factors influencing community assembly and the overall function of these symbiont communities.     

Effects of an introduced crayfish, Procambarus clarkii, on the distribution of south-western Iberian amphibians in their breeding habitats

The introduction of exotic species in aquatic habitats is one of the causes for the amphibian declines observed worldwide. In the 1970s, the red swamp crayfish Procambarus clarkii was introduced in the southwest Iberian Peninsula, where no native crayfish occur. In this study we assess the effect of P. clarkii presence in the breeding site distribution of each of the 13 southwest Iberian amphibians, while simultaneously accounting for the effects of potentially confounding habitat variables, as well as for the effects of the other large aquatic predators in the study area – predatory fish. Amphibian species richness was lower in places where P. clarkii was present than in places without P. clarkii , regardless of fish presence. After accounting for habitat variables and fish, crayfish presence was a negative predictor of the breeding probability for all urodeles ( Pleurodeles waltl , Salamandra salamandra , Triturus boscai and T. marmoratus ) and for two anurans ( Pelobates cultripes and Bufo bufo ). The majority of the species affected usually breed in temporary ponds without fish, but that may be colonized by the crayfish. The ongoing expansion of P. clarkii may eventually lead to a growing isolation of amphibian populations and ultimately to local extinctions and a permanent alteration of the amphibian communities in southwest Iberian Peninsula.     

Optimizing protection efforts for amphibian conservation in Mediterranean landscapes

Amphibians epitomize the modern biodiversity crisis, and attract great attention from the scientific community since a complex puzzle of factors has influence on their disappearance. However, these factors are multiple and spatially variable, and declining in each locality is due to a particular combination of causes. This study shows a suitable statistical procedure to determine threats to amphibian species in medium size administrative areas. For our study case, ten biological and ecological variables feasible to affect the survival of 15 amphibian species were categorized and reduced through Principal Component Analysis. The principal components extracted were related to ecological plasticity, reproductive potential, and specificity of breeding habitats. Finally, the factor scores of species were joined in a presence-absence matrix that gives us information to identify where and why conservation management are requires. In summary, this methodology provides the necessary information to maximize benefits of conservation measures in small areas by identifying which ecological factors need management efforts and where should we focus them on.     

甲状腺激素在两栖动物变态过程中的作用

两栖动物的幼体变态是研究甲状腺激素调节组织和器官重构的理想模式。本文主要综述了近年来两栖动物甲状腺激素合成过程中3种脱碘酶D1、D2和D3的特点及其生物学功能;甲状腺激素受体的蛋白结构、类型和机能;以及甲状腺激素对两栖动物幼体变态过程中各个类型组织和器官重构的调节;甲状腺激素、甲状腺激素受体和脱碘酶的互作,并展望了今后的研究方向。     

Complex impact of an invasive crayfish on freshwater food webs

Invasive alien species can have complex effects on native ecosystems, and interact with multiple components of food webs, making it difficult a comprehensive quantification of their direct and indirect effects. We evaluated the relationships between the invasive crayfish, Procambarus clarkii, amphibian larvae and predatory insects, to quantify crayfish impacts on multiple levels of food webs, and to evaluate whether crayfish predation of aquatic insects has indirect consequences for their preys. We used pipe sampling to assess the abundance of crayfish, amphibian larvae and their major predators (Ditiscidae, Notonectidae and larvae of Anisoptera) in invaded and uninvaded ponds within a human dominated landscape. We disentangled the multivariate effects of P. clarkii on different components of food web through a series of constrained redundancy analyses. The crayfish had a negative, direct impact on both amphibian communities and their predators. Amphibian abundance was negatively related to both predators. However, the negative, direct effects of crayfish on amphibians were much stronger than predation by native insects. Our results suggest that this crayfish impacts multiple levels of food webs, disrupting natural prey-predator relationships.     

Elevational differences in trait response to UV-B radiation by long-toed salamander populations

Amphibian species capable of optimizing trait response to environmental stressors may develop complex strategies for defending against rapid environmental change. Trait responses may differ between populations, particularly if stressor strength varies across spatial or temporal gradients. Ultraviolet-B (UV-B) radiation is one such stressor that poses a significant threat to amphibian species. We examined the ability of long-toed salamanders (Ambystoma macrodactylum) at high- and low-elevation breeding sites to cooperatively employ behavioral and physiological trait responses to mediate UV-B damage. We performed a microhabitat survey to examine differences in oviposition behavior and UV-B conditions among breeding populations at high- (n = 3; >1,500 m) and low-elevation (n = 3; <100 m) sites. We found significant differences in oviposition behavior across populations, with females at high-elevation sites selecting oviposition substrates in UV-B protected microhabitats. We also collected eggs (n = 633) from each of the breeding sites for analysis of photolyase activity, a photoreactivating enzyme that repairs UV-B damage to the DNA, using a photoproduct immunoassay. Our results revealed no significant differences in photolyase activity between long-toed salamander populations at high and low elevations. For high-elevation salamander populations, relatively low physiological repair capabilities in embryos appear to be buffered by extensive behavioral modifications to reduce UV-B exposure and standardize developmental temperatures. This study provides valuable insight into environmental stress responses via the assessment of multiple traits in allowing sensitive species to persist in rapidly changing landscapes.     

Kasstasin: A novel potent vasoconstrictor peptide from the skin secretion of the African red-legged running frog, Kassina maculata

Amphibian skin secretions are established sources of bioactive peptides. Here we describe the isolation, structural and pharmacological characterisation of a novel vasoconstrictor peptide from the skin secretion of the African hyperoliid frog, Kassina maculata, which exhibits no structural similarity to any known class of amphibian skin peptide. The peptide consists of 21 amino acid residues, FIKELLPHLSGIIDSVANAIK, and is C-terminally amidated. The provisional structure was obtained by MS/MS fragmentation using an Orbitrap mass spectrometer and L/I ambiguities were resolved following molecular cloning of biosynthetic precursor-encoding cDNA. A synthetic replicate of the peptide was found to possess weak antimicrobial and haemolytic activities but was exceptionally effective in constricting the smooth muscle of rat tail artery (EC₅₀ of 25pM). In reflection of its exceptional potency in constricting rat arterial smooth muscle, the peptide was named kasstasin, a derivation of Kassina and “stasis” (stoppage of flow). These data illustrate the continuing potential of amphibian skin secretions to provide novel natural peptide templates for biological evaluation.     

The distribution of amphibian species richness in protected areas of Chiapas,Mexico

Due to the current environmental crisis, many animal species face extinction problems. Amphibian populations have been affected by this crisis. Our goal is to study amphibian species diversity in Chiapas, which has 7.6% of the endemic amphibians in Mexico and 53 protected areas. Only 58% of the protected areas have management plans or information on their resident amphibians. We aim to determine the extent of protection provided by the network of natural areas for the conservation of amphibian species in the state and to discuss the effectiveness of this protection. Therefore, we compiled a georeferenced database of 112 amphibian species in Chiapas to create each distribution model. In addition, we carried out representativeness, beta diversity, and species richness analyses. As a result, we obtained a high degree of representativeness for the records and species distribution models. However, we found a decrease in the richness of amphibians involving 20% of total species, 13% of endemics, 18% threatened according to NOM-059, and 31% threatened according to IUCN between 1800 and 2020 and 1980–2020. We also identified two biodiversity hotspots in the Sierra Madre de Chiapas and the Northern Highlands physiographic regions. Finally, based on potential distributions, we found more endemic and threatened species outside protected natural areas than inside them. Our results give a broader picture of how amphibian richness is distributed in Chiapas. This information can help to prioritize conservation efforts toward those areas rich in threatened or endemic species, such as the Northern Mountains Hotspot we identified in northern Chiapas.     

Amphibian and reptile communities of rainforest fragments: minimum patch size to support high richness and abundance

Information on animal communities inhabiting Neotropical fragmented landscapes is important for developing conservation strategies. The structure of amphibian and reptile communities in six tropical rainforest fragments (<20?ha) and two reference areas in continuous forest at Los Tuxtlas, Mexico was studied. A total of 3,481 individuals of 51 species of amphibians and reptiles were recorded across 12 bimonthly surveys during 2 years. Taxonomic composition was different between the smallest fragments and the reference areas. Six species were exclusive to large undisturbed forest and richness was significantly lower in the five smallest fragments (1.4–6.6?ha) compared with the largest patch, one or both of the reference areas. Amphibian abundance tended to be higher in large areas, while reptiles were more abundant in the five smallest fragments. Craugastor loki and Anolis uniformis were the dominant species in all sites, and particularly in the smaller fragments. Amphibian and reptile richness was positively related to larger patch sizes, deeper leaf litter, closed canopy cover, and higher relative humidity and negatively related to linear patch shape and high temperatures. Abundance of reptiles was positively associated to high temperatures, high density of woody debris, and closed canopy cover; it was negatively affected by linear patch shape, low humidity levels, and steeper slopes. While amphibian and reptile communities were strongly affected in vegetation fragments, these patches retained a considerable number of rainforest species. Fragments up to at least 17?ha have the potential for preserving communities with similar structure to those occurring in large tropical rainforests.     

Dispersal and the metapopulation paradigm in amphibian ecology and conservation: are all amphibian populations metapopulations?

Amphibians are frequently characterized as having limited dispersal abilities, strong site fidelity and spatially disjunct breeding habitat. As such, pond‐breeding species are often alleged to form metapopulations. Amphibian species worldwide appear to be suffering population level declines caused, at least in part, by the degradation and fragmentation of habitat and the intervening areas between habitat patches. If the simplification of amphibians occupying metapopulations is accurate, then a regionally based conservation strategy, informed by metapopulation theory, is a powerful tool to estimate the isolation and extinction risk of ponds or populations. However, to date no attempt to assess the class‐wide generalization of amphibian populations as metapopulations has been made. We reviewed the literature on amphibians as metapopulations (53 journal articles or theses) and amphibian dispersal (166 journal articles or theses for 53 anuran species and 37 salamander species) to evaluate whether the conditions for metapopulation structure had been tested, whether pond isolation was based only on the assumption of limited dispersal, and whether amphibian dispersal was uniformly limited. We found that in the majority of cases (74%) the assumptions of the metapopulation paradigm were not tested. Breeding patch isolation via limited dispersal and/or strong site fidelity was the most frequently implicated or tested metapopulation condition, however we found strong evidence that amphibian dispersal is not as uniformly limited as is often thought. The frequency distribution of maximum movements for anurans and salamanders was well described by an inverse power law. This relationship predicts that distances beneath 11–13 and 8–9 km, respectively, are in a range that they may receive one emigrating individual. Populations isolated by distances approaching this range are perhaps more likely to exhibit metapopulation structure than less isolated populations. Those studies that covered larger areas also tended to report longer maximum movement distances – a pattern with implications for the design of mark‐recapture studies. Caution should be exercised in the application of the metapopulation approach to amphibian population conservation. Some amphibian populations are structured as metapopulations – but not all.     

Senegalin: a novel antimicrobial/myotropic hexadecapeptide from the skin secretion of the African running frog, Kassina senegalensis

Amphibian skin is a rich and unique source of novel bioactive peptides most of which are endowed with either antimicrobial or pharmacological properties. Here, we report the identification and structural characterization of a novel peptide, named senegalin, which possesses both activities. Senegalin is a hexadecapeptide amide (FLPFLIPALTSLISSL-NH₂) of unique primary structure found in the skin secretion of the African running frog, Kassina senegalensis. The structure of the biosynthetic precursor of senegalin, deduced from cloned skin cDNA, consists of 76 amino acid residues and displays the typical domain organization of an amphibian skin peptide precursor. Both natural senegalin and its synthetic replicate displayed antimicrobial and myotropic activities. Senegalin was active against Staphylococcus aureus (MIC 50 μM) and Candida albicans (MIC 150 μM) but was non-haemolytic at concentrations up to and including 150 μM. In contrast, senegalin induced a dose-dependent contraction of rat urinary bladder smooth muscle (EC₅₀ 2.9 nM) and a dose-dependent relaxation of rat tail artery smooth muscle (EC₅₀ 37.7 nM). Senegalin thus represents a prototype biologically active amphibian skin peptide and illustrates the fact that amphibian skin secretion peptidomes continue to be unique sources of such molecules.     

Gene cloning and characterization of novel antinociceptive peptide from the brain of the frog, Odorrana grahami

Amphibian opiate peptides including dermorphins and deltorpins have been recently found only in the skin of South American frogs belonging to the subfamily Phyllomedusinae (Phyllomedusa, Agalychnis and Pachymedusa species). No opiate peptides have ever been identified from other amphibians or organs except skin. Here we report the purification and characterization of a novel antinociceptive peptide named odorranaopin from the homogenates of the frog brains, Odorrana grahami, which is also the first antinociceptive peptide found in Ranidae amphibian. Odorranaopin comprises 17 amino acid residues with the sequence of DYTIRTRLHQESSRKVL (Mr 2102 Da). The cDNA encoding odorranaopin was cloned from the frog brain cDNA library, and it was confirmed to be a specific gene. The odorranaopin precursor deduced is composed of 61 amino acid residues including the predicted signal peptide, acidic spacer peptide and mature odorranaopin positioned at the C-terminus. Odorranaopin could inhibit nociceptive responses induced by formalin and acetic acid. It also inhibited the contractile responses of ileum smooth muscle induced by bradykinin, implying that the antinociceptive activity of odorranaopin possibly results from its blockade on bradykinin or bradykinin receptor functions. Odorranaopin is the first antinociceptive peptide found in Ranidae amphibian.     

Climate change correlates with rapid delays and advancements in reproductive timing in an amphibian community

Climate change has had a significant impact globally on the timing of ecological events such as reproduction and migration in many species. Here, we examined the phenology of reproductive migrations in 10 amphibian species at a wetland in South Carolina, USA using a 30 year dataset. We show for the first time that two autumn-breeding amphibians are breeding increasingly later in recent years, coincident with an estimated 1.2°C increase in local overnight air temperatures during the September through February pre-breeding and breeding periods. Additionally, two winter-breeding species in the same community are breeding increasingly earlier. Four of the 10 species studied have shifted their reproductive timing an estimated 15.3 to 76.4 days in the past 30 years. This has resulted in rates of phenological change that range from 5.9 to 37.2 days per decade, providing examples of some of the greatest rates of changing phenology in ecological events reported to date. Owing to the opposing direction of the shifts in reproductive timing, our results suggest an alteration in the degree of temporal niche overlap experienced by amphibian larvae in this community. Reproductive timing can drive community dynamics in larval amphibians and our results identify an important pathway by which climate change may affect amphibian communities.     

Metacommunity dynamics of amphibians in years with differing rainfall

Many studies investigated the habitat preference and behaviour ecology of individual amphibian species while we know less about how their community assembly reflects changes in environmental factors, including the role of climatic extremes. Community-level studies also allow us to apply trait-based analyses that are crucial for a better understanding of the functioning of amphibian communities and metacommunities. In two years with contrasting rainfall (2012 and 2013), we found amphibian species in 85 different waterbodies of a heterogeneous landscape in Central Europe (Hungary). Within the metacommunity framework, the contributions of local, landscape and spatial variables to community assembly were assessed. We also measured the local extinction–colonisation rates in the ponds for all species between the two years. To investigate the role of dispersal traits in explaining the spatial distribution of species, we studied the relationship between body size and the pure spatial fraction of variation. According to our results, the main drivers were the same in both the dry and wet year, but their relative contribution changed. Local variables played a predominant role in the assembly of the amphibian metacommunity. Spatial signals were more evident in the dry year. This implies not only the adverse effect of decreased connectivity due to the drying out of several habitats but also a loss of breeding sites for the studied amphibians. Local colonisation rates were higher in primarily terrestrial species (Hyla arborea, Pelobates fuscus, Bufo bufo) which only visit ponds during breeding. We found a negative relationship between the pure spatial effect and body size, suggesting an increased level of dispersal limitation in small-bodied species. Our results showed that while the strength and relative role of local and spatial processes changed between years, the role of dispersal traits in explaining the spatial distribution of species was similar. Specialisation to different habitats seems to be a major process in determining vertebrate metacommunities in landscapes. Dispersal traits of different species should be taken more into consideration in the practical conservation of amphibian habitats.