This study aims to analyze the thermal biology and climatic vulnerability of two closely related lizard species (Stenocercus festae and S. guentheri) inhabiting the Ecuadorian Andes at high altitudes. Four physiological parameters—body temperature (Tb), preferred temperature (Tpref), critical thermal maximum (CTmax), and critical thermal minimum (CTmin)—were evaluated to analyze the variation of thermophysiological traits among these populations that inhabit different environmental and altitudinal conditions. We also evaluate the availability of operative temperatures, warming tolerance, and thermal safety margin of each population to estimate their possible risks in the face of future raising temperatures. Similar to previous studies, our results suggest that some physiological traits (CTmax and Tb) are influenced by environmental heterogeneity, which brings changes on the thermoregulatory behavior. Other parameters (Tpref and CTmin), may be also influenced by phylogenetic constraints. Moreover, the fluctuating air temperature (Tair) as well as the operative temperatures (Te) showed that these lizards exploit a variety of thermal microenvironments, which may facilitate behavioral thermoregulation. Warming tolerance and thermal safety margin analyses suggest that both species find thermal refugia and remain active without reducing their performance or undergoing thermal stress within their habitats. We suggest that studies on the thermal biology of tropical Andean lizards living at high altitudes are extremely important as these environments exhibit a unique diversity of microclimates, which consequently result on particular thermophysiological adaptations.

Captive-bred organisms are widely used in ecology, evolution and conservation research, especially in scenarios where natural populations are scarce or at risk of extinction. Yet, it is still unclear whether captivity may alter thermal tolerances, crucial traits to predict species resilience to global warming. Here, we study whether captive-bred tadpoles of the gliding treefrog (Agalychnis spurrelli) show different thermal tolerances than wild-caught individuals. Our results show that there are no differences between critical thermal limits (CTmax and CTmin) of captive-bred and wild-caught tadpoles exposed to three-day acclimatization at 20 °C. Therefore, we suggest that the use of captive-bred amphibians is valid and may be appropriate in experimental comparisons to thermal physiological studies of wild populations.

Ecuador’s territory harbors a unique set of species and ecosystems, many of them endemic to the countries’ territory and subject to different sources of threat of anthropogenic origin. Despite national and subnational conservation strategies developed in Ecuador to conserve its biodiversity in the long run, including the National System of Protected Areas (PANE) and the forest conservation incentive program SocioBosque (PSB), further actions are needed to mitigate and reverse the effects of threats for the persistence of biodiversity. This study was designed to identify the most important areas for biodiversity conservation in mainland Ecuador that can contribute to preserving key species (i.e. endemic, threatened) and ecosystems in the wider landscape, thus complementing current conservation efforts (i.e. PANE). Species distribution models and recent maps were used to identify a set of 744 species and 87 ecosystems as surrogates of the country’s biodiversity. Marxan, a systematic reserve selection algorithm was used to identify important biodiversity areas that could represent between 10% and 20% of the remnant distribution of the surrogates. The optimized solution generated by Marxan included 24% (3.64 million ha) of Ecuador′s remnant vegetation, of which 35% is within the current national protected area system and 13% (456 000 ha) are included within SocioBosque communal and private conservation agreements. Major conservation shortfalls of the PANE were concentrated in the Southern Andes, Central Amazonia, and the Central and Southern portions of the Coastal plain.

This study aims to analyze the thermal biology and climatic vulnerability of two closely related lizard species (Stenocercus festae and S. guentheri) inhabiting the Ecuadorian Andes at high altitudes. Four physiological parameters—body temperature (Tb), preferred temperature (Tpref), critical thermal maximum (CTmax), and critical thermal minimum (CTmin)—were evaluated to analyze the variation of thermophysiological traits among these populations that inhabit different environmental and altitudinal conditions. We also evaluate the availability of operative temperatures, warming tolerance, and thermal safety margin of each population to estimate their possible risks in the face of future raising temperatures. Similar to previous studies, our results suggest that some physiological traits (CTmax and Tb) are influenced by environmental heterogeneity, which brings changes on the thermoregulatory behavior. Other parameters (Tpref and CTmin), may be also influenced by phylogenetic constraints. Moreover, the fluctuating air temperature (Tair) as well as the operative temperatures (Te) showed that these lizards exploit a variety of thermal microenvironments, which may facilitate behavioral thermoregulation. Warming tolerance and thermal safety margin analyses suggest that both species find thermal refugia and remain active without reducing their performance or undergoing thermal stress within their habitats. We suggest that studies on the thermal biology of tropical Andean lizards living at high altitudes are extremely important as these environments exhibit a unique diversity of microclimates, which consequently result on particular thermophysiological adaptations.

Necrophilia in amphibians is a poorly known behaviour despite its potential as a beneficial adaptation for improving reproductive success. Here, we describe the observation of a multiple amplexus involving necrophilia in the recently described Tsachila snouted treefrog, Scinax tsachila (Anura: Hylidae). We further provide an extensive review of published necrophilia in amphibians. At least 33 species of amphibians, mostly anurans, have shown a necrophiliac behaviour, with only one case of necrophilia in a caudate. Necrophilia has long been considered a maladaptive behaviour, since reproduction is usually not viable and is also associated with increased risk of death. However, necrophiliac behaviour has recently been proposed as an adaptive behaviour for some species because it may result in viable offspring.

Microbiome–pathogen interactions are increasingly recognized as an important element of host immunity. While these host-level interactions will have consequences for community disease dynamics, the factors which influence host microbiomes at larger scales are poorly understood. We here describe landscape-scale pathogen–microbiome associations within the context of post-epizootic amphibian chytridiomycosis, adisease caused by the panzootic chytrid fungus Batrachochytrium dendrobatidis. We undertook a survey of Neotropical amphibians across altitudinal gradients in Ecuador~30 years following the observed amphibian declines and collected skin swab-sam-ples which were metabarcoded using both fungal (ITS-2) and bacterial (r16S) ampli-cons. The data revealed marked variation in patterns of both B. dendrobatidis infection and microbiome structure that are associated with host life history. Stream breeding amphibians were most likely to be infected with B. dendrobatidis. This increased probability of infection was further associated with increased abundance and diversity of non-Batrachochytrium chytrid fungi in the skin and environmental microbiome.We also show that increased alpha diversity and the relative abundance of fungi are lower in the skin microbiome of adult stream amphibians compared to adult pond breeding amphibians, an association not seen for bacteria. Finally, stream tadpoles exhibit lower proportions of predicted protective microbial taxa than pond tadpoles,suggesting reduced biotic resistance. Our analyses show that host breeding ecology strongly shapes pathogen–microbiome associations at a landscape scale, a trait that may influence resilience in the face of emerging infectious diseases.

Aim: We analysed elevational and microclimatic drivers of thermal tolerance diversity in a tropical mountain frog clade to test three macrophysiological predictions: less spatial variation in upper than lower thermal limits (Bretts’ heat‐invariant hypoth‐ esis); narrower thermal tolerance ranges in habitats with less variation in temperature (Janzen's climatic variability hypothesis); and higher level of heat impacts at lower elevations. Location: Forest and open habitats through a 4,230‐m elevational gradient across the tropical Andes of Ecuador. Method: We examined variability in critical thermal limits (CTmax and CTmin) and thermal breadth (TB; CTmax–CTmin) in 21 species of Pristimantis frogs. Additionally, we monitored maximum and minimum temperatures at the local scale (tmax, tmin) and estimated vulnerability to acute thermal stress from heat (CTmax–tmax) and cold (tmin–CTmin), by partitioning thermal diversity into elevational and microclimatic variation. Results: Our results were consistent with Brett's hypothesis: elevation promotes more variation in CTmin and tmin than in CTmax and tmax. Frogs inhabiting thermally variable open habitats have higher CTmax and tmax and greater TBs than species restricted to forest habitats, which show less climatic overlap across the elevational gradient (Janzen's hypothesis). Vulnerability to heat stress was higher in open than forest habitats and did not vary with elevation.

Bone alterations due to metabolic bone disease in captive animal populations can have a negative impact on repopulation and research initiatives. This investigation has the purpose of describing the principal radiographic and anatomopathological findings present in nine gliding leaf frogs (Agalychnis spurrelli) kept in captivity with alterations in their spines and long bones. The observed histopathological findings were in the canalis vertebralis, paraspinal muscle and long bones, and included deformed bones with alteration of the adjacent tissues, alterations in the ossification process, bone degeneration and resorption, decreased number of osteocytes and deposition of osteoid and fibrous material in the compact bone tissue. Additionally, the spinal cord showed compressed white matter, chronic meningitis in the duramater, alteration in the number of glial cells and loss of delimitation between the gray and white matter. Radiographical changes were found mainly in the long bones and included moth-eaten osteolysis, solid periosteal reaction, bone deformities, cortical tunneling and inflammation of adjacent soft tissues. Also, pathological fractures of the femur and urostyle were observed together with spinal column deviations with increased bone density.

The Andes have experienced an unprecedented wave of amphibian declines and extinctions that are linked to a combination of habitat reduction and the spread of the fungal pathogen, Batrachochytrium dendrobatidis (Bd). In the present study, a range of high-altitude habitats in Southern Ecuador were surveyed for the presence of Bd. With a particular focus on Yacuri National Park, infection data are presented from across the resident amphibian community. This community contains a once putatively extinct species which was rediscovered in 2016, the Podocarpus Stubfoot Toad (Atelopus podocarpus). Across species, local Bd prevalence was 73% in tadpoles (n = 41 individuals from three species) and 14% in adults (n = 43 individuals from 14 species). Strikingly, 93% (14/15) of tested tadpoles of the recently described local endemic, Gastrotheca yacuri, were infected with a high pathogen load, suggesting that this species likely acts as a reservoir of infection in Yacuri. These fndings show that the threat of disease for A. podocarpus still exists, and that this species requires urgent action to ensure its survival

Amphibian skin is highly variable in structure and function across anurans, and plays an important role in physiological homeostasis and immune defense. For example, skin sloughing has been shown to reduce pathogen loads on the skin, such as the lethal fungus Batrachochytrium dendrobatidis (Bd), but interspecific variation in sloughing frequency is largely unknown. Using phylogenetic linear mixed models, we assessed the relationship between skin turnover rate, skin morphology, ecological traits and overall evidence of Bd-driven declines. We examined skin sloughing rates in 21 frog species from three continents, as well as structural skin characteristics measured from preserved specimens. We found that sloughing rate varies significantly with phylogenetic group, but was not associated with evidence of Bd-driven declines, or other skin characteristics examined. This is the first comparison of sloughing rate across a wide range of amphibian species, and creates the first database of amphibian sloughing behavior. Given the strong phylogenetic signal observed in sloughing rate, approximate sloughing rates of related species may be predicted based on phylogenetic position. While not related to available evidence of declines, understanding variation in sloughing rate may help explain differences in the severity of infection in genera with relatively slow skin turnover rates (e.g. Atelopus)

Perhaps the most well-studied aspect of anuran communication, vocalizations play an integral role in mate attraction (Littlejohn 1977; Gerhardt 1991; Tobias et al. 1998), demarcation of territory (Wagner 1989; Byrne 2009), and myriad other social interactions (Wells 1977; Wells and Schwartz 2007). Environmental factors such as temperature and humidity, as well as individual intrinsic factors such as size, are known to cause variations in call frequency and duration (Lingnau and Bastos 2007; De Toledo et al. 2009; Costa and Toledo 2013). However, the majority of variation is caused by changes in behavioral context. The genus Pristimantis (Craugastoridae) has undergone extensive taxonomic revision and contains ca. 530 currently recognized species (Hedges et al. 2008; Frost 2019). As with most members of this group, species of Pristimantis are highly cryptic, reclusive, and almost exclusively nocturnal (Lynch and Duellman 1997). For this reason, species data on calls and behavior are sparse (Duellman and Lehr 2009).

Chytrid fungi and viruses within the genus Ranavirus have been associated with mass mortality events and declines in amphibian populations worldwide. The fungus Batrachochytrium dendrobatidis (Bd) was reported in Ecuador; however, other chytrid fungi like Batrachochytrium salamandrivorans (Bsal) or ranaviruses have not been described in the country so far. To prevent the introduction of pathogens into amphibian populations under conservation programs and to implement a successful disease surveillance program, the development of a sensitive and specific diagnostic assay was required. We describe here the optimization of one TaqMan probe-based multiplex quantitative polymerase chain reaction (qPCR) assay that enables the simultaneous detection of Bsal and ranavirus, and one monoplex TaqMan qPCR assay for the detection of Bd. Standard curves, with a high linear correlation (r2 > 0.995), were generated using a synthetic genome template (gBlocks®) containing the target sequences from all three pathogens. Different samples from skin, liver, kidney, spleen, and lung from six different amphibian species were tested, and both qPCR assays showed highly reproducible and reliable results. To our knowledge, this method is the first multiplex qPCR system developed in Ecuador for identifying amphibian pathogens and represents a valuable tool for the early detection of these pathogens and for infection and co-infection monitoring in future epidemiological surveillance of amphibian species at risk of extinction.

Objective: Edema syndrome is highly prevalent but under researched in captive frogs around the world. The objective of the present study was to characterize at a basic microbiological and cytological level of the bacteria of the edema fuid of 20 individuals of the genus Gastrotheca to determine the presence of possible anaerobic and aerobic bacteria. Results: Fourteen types of bacteria were identified in the edema fuid, 12 of them at the species level (Pasteurella haemolytica, Hafnia alvei, Enterobacter agglomerans, Aeromonas hydrophila, Pseudomonas fuorescens, Burkholderia pseudomallei, Salmonella arizonae, Enterobacter gergoviae, Enterobacter sakazakii, Yersinia enterocolitica, Klebsiella oxytoca, and Klebsiella ozaenae) and two at the genus level (Enterococcus spp. and Streptococcus spp.). The most fre‑ quently identifed cells were lymphocytes (37.7% in females and 46.4% in males), erythrocytes (23.5% in females and 17.5% in males) and neutrophils (4.2% in females and 2.8% in males). Finally, no relationship was found between the data obtained and the sex of the individuals studied.

Here, we describe the extreme plasticity in colour variation in the tadpole of the executioner clownfrog, Dendropsophus carnifex (Anura: Hylidae), found across nearby ponds in Mindo (Ecuador). Tadpole coloration was compared between individuals from four distinct ponds revealing two explicit colour schemes, a pale phenotype which is commonly described for this species and a bright phenotype which, to our knowledge, has not been described before. In addition, the bright phenotype revealed reversibility to the pale phenotype in laboratory conditions. We discuss the functionality and driving factors for these contrasting colour phenotypes including the potential role of distinct ecological conditions.

Many amphibian species are at risk of extinction in their natural habitats due to the presence of the fungal pathogen Batrachochytrium dendrobatidis (Bd). For the most highly endangered species, captive assurance colonies have been established as an emergency measure to avoid extinction. Experimental research has suggested that symbiotic microorganisms in the skin of amphibians play a key role against Bd. While previous studies have addressed the effects of captivity on the cutaneous bacterial community, it remains poorly studied whether and how captive conditions affect the proportion of beneficial bacteria or their anti-Bd performance on amphibian hosts. In this study we sampled three amphibian species of the highly threatened genus, Atelopus, that remain in the wild but are also part of ex situ breeding programs in Colombia and Ecuador. Our goals were to (1) estimate the diversity of culturable bacterial assemblages in these three species of Atelopus, (2) describe the effect of captivity on the composition of skin microbiota, and (3) examine how captivity affects the bacterial ability to inhibit Bd growth. Using challenge assays we tested each bacterial isolate against Bd, and through sequencing of the 16S rRNA gene, we identified species from thirteen genera of bacteria that inhibited Bd growth. Surprisingly, we did not detect a reduction in skin bacteria diversity in captive frogs.