Biomarkers at the Individual and Biochemical Level: Effects of Pure and Formulated Lambda-Cyhalothrin in Boana pulchella Tadpoles (Duméril and Bibron, 1841).

We compared the effects of lambda-cyhalothrin as the pure active ingredient and as a formulated product (Zero®), on the larval stage of the autochthonous species Boana pulchella. We evaluated ecotoxicological endpoints, behavioral and developmental alterations, and the biochemical detoxifying, neurotoxic, and oxidative stress responses, covering a wide concentration range from environmental to high application levels. Both pyrethroid preparations displayed similar ecotoxicity (median lethal concentration of ~0.5 mg/L), with the lethal effect of Zero® being more pronounced than that of the active ingredient. Sublethal behavioral alterations in natatory activity were observed at 1000 times lower concentrations, indicating the ecological hazard of tadpole exposure to this pyrethroid at environmentally relevant concentrations. Biochemical endpoints in B. pulchella larvae showed significant responses to lambda-cyhalothrin in the ng/L range; these responses were different for the pure or the formulated product, and they were variable at higher concentrations. Principal components analysis confirmed the prevalence of biochemical responses as early endpoints at the lowest lambda-cyhalothrin concentrations; the Integrated Biomarker Response Index proportionally increased with pyrethroid concentration in a similar way for the pure and the formulated products. We conclude that lambda-cyhalothrin is of concern from an environmental perspective, with particular emphasis on autochthonous anuran development. The battery of biochemical biomarkers included in our study showed a consistent integrated biomarker response, indicating that this is a potent tool for monitoring impacts on amphibians. Environ Toxicol Chem 2024;00:1-11. © 2024 SETAC.

Ticks parasitizing cold-blooded animals from three different Brazilian biomes; with note for males of Amblyomma rotundatum.

Ticks parasitize a wide variety of wild animals, including amphibians and reptiles. In addition to the possibility of microorganism transmission to these hosts, ticks can also cause severe bleeding, and high parasitism can lead to death. Therefore, knowing the diversity of ticks parasitizing amphibians and reptiles is important for conservation and preservation measures for these vertebrates. In the present study, we report parasitism by ticks in amphibians and reptiles from different Brazilian biomes (Amazon, Caatinga, Cerrado and Atlantic Forest). Ticks were collected from amphibians and reptiles deposited from the Herpetological Collection of the Federal University of Maranhão (UFMA), São Luís (Maranhão State), the State University Santa Cruz (UESC), Ilhéus (Bahia State), and the Federal University of São Francisco Valley (Univasf), Petrolina (Pernambuco State). Additionally, ticks were collected from amphibians and reptiles captured and road-killed in the Amazon biome, at Maranhão and Amapá States. Specimens of ticks were photographed under a Zeiss stereomicroscope (5.1 zoom). Map with the locations were made using the Qgis program. Overall, 1973 specimens of amphibians and reptiles were examined. A total of 927 ticks were collected: 98 larvae, 421 nymphs and 408 adults. Six species of ticks were identified: Amblyomma rotundatum and Amblyomma dissimile the most frequent, and Amblyomma cajennense sensu stricto, Amblyomma sculptum, Amblyomma nodosum and Amblyomma humerale, occasionally. Surprisingly, a total of twelve males of A. rotundatum were collected. Here we report new records of association between cold-blooded animals and ticks and reinforce the absence of A. dissimile in the Caatinga, Cerrado and Atlantic Forest biomes. Additionally, we report new records of A. rotundatum males on reptiles in the Amazon biome. This last record allows us to speculate about a possible association of A. rotundatum males with reptiles and the Amazon biome.

Food web collapse and regime shift following goldfish introduction in permanent ponds.

In a global context of invasive alien species (IAS), native predators are often eradicated by functionally different IAS, which may induce complex cascading consequences on ecosystem functioning because of the key role predators play in structuring communities and stabilizing food webs. In permanent ponds, the most abundant freshwater systems on Earth, global human-mediated introductions of alien omnivores such as the pet trade goldfish are driving broad-scale patterns of native predators' exclusion, but cascading consequences on food web structure and functioning are critically understudied. We compared food webs of naturally fishless ponds versus ponds where dominant native predators (newts) had been extirpated by invasive goldfish within the last decade. Integrating community-wide isotopic, taxonomic and functional traits approaches, our study reveals that pond food webs collapsed in both vertical and horizontal dimensions following goldfish introduction and the associated exclusion of native predators. Consumer taxonomic diversity was drastically reduced, essentially deprived of amphibians as well as predatory and mobile macroinvertebrates to the profit of burrowing, lower trophic level consumers (detritivores). Changes in community structure and function underlined a regime shift from a macrophyte-dominated system mainly characterized by benthic primary production (periphyton), to a macrophyte-depleted state of ponds hosting communities mainly associated with phytoplankton primary production and detritus accumulation, with higher tolerance to eutrophication and low dissolved oxygen concentration. Results underline major impacts of widely introduced omnivores such as the goldfish on the functioning of pond ecosystems with potentially dramatic consequences on the key ecosystem services they deliver, such as global biodiversity support or water quality improvement. They also shed light on the key role of submerged aquatic vegetation in supporting diverse communities and complex food webs in shallow lentic systems and call for urgent consideration of threats posed by IAS on ponds' ecosystems by managers and policymakers.

Dynamics of Amphibian Pathogen Detection Using Extended Museum Specimens.

Natural history collections have long served as the foundation for understanding our planet's biodiversity, yet they remain a largely untapped resource for wildlife disease studies. Extended specimens include multiple data types and specimen preparations that capture the phenotype and genotype of an organism and its symbionts-but preserved tissues may not always be optimized for downstream detection of various pathogens. Frogs are infected by an array of pathogens including Batrachochytrium dendrobatidis (Bd), Ranavirus (Rv), and Amphibian Perkinsea (Pr), which provides the opportunity to study differences in detection dynamics across tissue types. We used quantitative PCR protocols to screen two tissue types commonly deposited in museum collections, toe clips and liver, from two closely related host species, Rana catesbeiana and Rana clamitans. We compared Bd, Rv, and Pr infection prevalence and intensity between species and tissue types and found no significant difference in prevalence between species, but Bd intensity was higher in R. clamitans than R. catesbeiana. Toe tissue exhibited significantly higher Bd infection loads and was more useful than liver for detecting Bd infections. In contrast, Rv was detected from more liver than toe tissues, but the difference was not statistically significant. Our results support the use of extended specimen collections in amphibian disease studies and demonstrate that broader tissue sampling at the time of specimen preparation can maximize their utility for downstream multipathogen detection.

Variation in the sensitivity of intestine and skin of Bufo gargarizans and Rana chensinensis tadpoles in relation to zinc exposure.

Zinc (Zn) contaminants in the aquatic environment have an intricate impact on amphibians. Amphibian gut and skin microbiota are participated in regulating their normal physiological functions. Here, we investigated the effects of Zn on the gut and skin tissues and microbiota of Bufo gargarizans and Rana chensinensis tadpoles using histological methods and 16S rRNA sequencing technology. Our results showed a decrease in the height of enterocytes and skin epithelial cells after Zn treatment. Furthermore, Zn exposure elicited alterations in the composition and structure of the gut and skin microbiota at the phylum and genus levels in Bufo gargarizans and Rana chensinensis tadpoles. The feature predictions revealed an elevation in the abundance of potentially pathogenic bacteria and stress-tolerant bacteria in the gut and skin of both tadpoles after zinc exposure. We also speculated that microbiota from various species and organs exhibit varying degrees of sensitivity to zinc based on the functional predictions results. In the context of increasing environmental pollution and the global amphibians decline, our research enriches the current understanding of effects of zinc on amphibian microbiota and provides new framework for artificial breeding and amphibian conservation.

The Amphibian Genomics Consortium: advancing genomic and genetic resources for amphibian research and conservation.

Amphibians represent a diverse group of tetrapods, marked by deep divergence times between their three systematic orders and families. Studying amphibian biology through the genomics lens increases our understanding of the features of this animal class and that of other terrestrial vertebrates. The need for amphibian genomics resources is more urgent than ever due to the increasing threats to this group. Amphibians are one of the most imperiled taxonomic groups, with approximately 41% of species threatened with extinction due to habitat loss, changes in land use patterns, disease, climate change, and their synergistic effects. Amphibian genomics resources have provided a better understanding of ontogenetic diversity, tissue regeneration, diverse life history and reproductive modes, antipredator strategies, and resilience and adaptive responses. They also serve as critical models for understanding widespread genomic characteristics, including evolutionary genome expansions and contractions given they have the largest range in genome sizes of any animal taxon and multiple mechanisms of genetic sex determination. Despite these features, genome sequencing of amphibians has significantly lagged behind that of other vertebrates, primarily due to the challenges of assembling their large, repeat-rich genomes and the relative lack of societal support. The advent of long-read sequencing technologies, along with computational techniques that enhance scaffolding capabilities and streamline computational workload is now enabling the ability to overcome some of these challenges. To promote and accelerate the production and use of amphibian genomics research through international coordination and collaboration, we launched the Amphibian Genomics Consortium (AGC) in early 2023. This burgeoning community already has more than 282 members from 41 countries (6 in Africa, 131 in the Americas, 27 in Asia, 29 in Australasia, and 89 in Europe). The AGC aims to leverage the diverse capabilities of its members to advance genomic resources for amphibians and bridge the implementation gap between biologists, bioinformaticians, and conservation practitioners. Here we evaluate the state of the field of amphibian genomics, highlight previous studies, present challenges to overcome, and outline how the AGC can enable amphibian genomics research to "leap" to the next level.

Impacts of cattle management and agricultural practices on water quality through different approaches: physicochemical and ecotoxicological parameters.

The intensification of livestock farming can pose risks to the environment due to the increased use of veterinary products and the generation of waste in confined areas. The quality of water bodies near livestock establishments (Areco River (A) and Doblado stream (D), San Antonio de Areco, Buenos Aires, Argentina) was studied by physicochemical parameters, metals, pesticides, emerging contaminants, and lethal and sublethal toxicity (neurotoxicity and oxidative stress) in larvae of the native amphibian Rhinella arenarum. Six sites were selected: upstream (S1A and S1D), at the level (S2A and S2D), and downstream (S3A and S3D) from the establishments. A low concentration of dissolved oxygen was observed in Doblado stream (< 2.34 mg/L). Cu, Mn, V, and Zn exceeded the limits for the protection of aquatic life at various sites. Between 24 and 34 pesticides were detected in all sites, with 2,4-D, atrazine, and metolachlor being the most recurrent. In water and sediment, the concentrations of ivermectin (S2A, 1.32 µg/L and 58.18 µg/kg; S2D, 0.8 µg/L and 85.22 µg/kg) and oxytetracycline (S2A, < 1 mg/L and < 1 mg/kg; S2D, 11.8 mg/L and 39 mg/kg) were higher at sites near the establishments. All sites caused between 30 and 38.3% of lethality and produced neurotoxicity and alterations in the reduced glutathione content. Moreover, larvae exposed to samples from all sites incorporated ivermectin. These results demonstrate the degradation of the studied sites in relation to the agricultural activities of the area, highlighting the need to take measures to protect and preserve aquatic ecosystems.

Using comparative extinction risk analysis to prioritize the IUCN Red List reassessments of amphibians.

Assessing the extinction risk of species based on the International Union for Conservation of Nature (IUCN) Red List (RL) is key to guiding conservation policies and reducing biodiversity loss. This process is resource demanding, however, and requires continuous updating, which becomes increasingly difficult as new species are added to the RL. Automatic methods, such as comparative analyses used to predict species RL category, can be an efficient alternative to keep assessments up to date. Using amphibians as a study group, we predicted which species are more likely to change their RL category and thus should be prioritized for reassessment. We used species biological traits, environmental variables, and proxies of climate and land-use change as predictors of RL category. We produced an ensemble prediction of IUCN RL category for each species by combining 4 different model algorithms: cumulative link models, phylogenetic generalized least squares, random forests, and neural networks. By comparing RL categories with the ensemble prediction and accounting for uncertainty among model algorithms, we identified species that should be prioritized for future reassessment based on the mismatch between predicted and observed values. The most important predicting variables across models were species' range size and spatial configuration of the range, biological traits, climate change, and land-use change. We compared our proposed prioritization index and the predicted RL changes with independent IUCN RL reassessments and found high performance of both the prioritization and the predicted directionality of changes in RL categories. Ensemble modeling of RL category is a promising tool for prioritizing species for reassessment while accounting for models' uncertainty. This approach is broadly applicable to all taxa on the IUCN RL and to regional and national assessments and may improve allocation of the limited human and economic resources available to maintain an up-to-date IUCN RL.

Uso del análisis comparativo del riesgo de extinción para priorizar la reevaluación de los anfibios en la Lista Roja de la UICN Resumen El análisis del riesgo de extinción de una especie con base en la Lista Roja (LR) de la Unión Internacional para la Conservación de la Naturaleza (UICN) es clave para guiar las políticas de conservación y reducir la pérdida de la biodiversidad. Sin embargo, este proceso demanda recursos y requiere de actualizaciones continuas, lo que se complica conforme se añaden especies nuevas a la LR. Los métodos automáticos, como los análisis comparativos usados para predecir la categoría de la especie en la LR, pueden ser una alternativa eficiente para mantener actualizados los análisis. Usamos a los anfibios como grupo de estudio para predecir cuáles especies tienen mayor probabilidad de cambiar de categoría en la LR y que, por lo tanto, se debería priorizar su reevaluación. Usamos las características biológicas de la especie, las variables ambientales e indicadores climáticos y del cambio de uso de suelo como predictores de la categoría en la LR. Elaboramos una predicción de ensamble de la categoría en la LR de la UICN para cada especie mediante la combinación de cuatro algoritmos diferentes: modelos de vínculo acumulativo, menor número de cuadros filogenéticos generalizados, bosques aleatorios y redes neurales. Con la comparación entre las categorías de la LR y la predicción de ensamble y con considerar la incertidumbre entre los algoritmos identificamos especies que deberían ser prioridad para futuras reevaluaciones con base en el desfase entre los valores predichos y los observados. Las variables de predicción más importantes entre los modelos fueron el tamaño de la distribución de la especie y su configuración espacial, las características biológicas, el cambio climático y el cambio de uso de suelo. Comparamos nuestra propuesta de índice de priorización y los cambios predichos en la LR con las reevaluaciones independientes de la LR de la UICN y descubrimos un buen desempeño tanto para la priorización como para la direccionalidad predicha de los cambios en las categorías de la LR. El modelo de ensamble de la categoría de la LR esa una herramienta prometedora para priorizar la reevaluación de las especies a la vez que considera la incertidumbre del modelo. Esta estrategia puede generalizarse para aplicarse a todos los taxones de la LR de la UICN y a los análisis regionales y nacionales. También podría mejorar la asignación de los recursos humanos y económicos limitados disponibles para mantener actualizada la LR de la UICN.

Assessment of Animal Welfare at an Exotic Animal Fair in Poland: A Focus on the Quality of Exhibition Containers for Reptiles and Amphibians.

Given the growing number of events involving exotic animals, it is crucial to prioritize the well-being of the animals involved. This study aims to evaluate the quality of animal presentation at a selected fair in Poland and assess the level of animal welfare evident in the exhibition boxes, contributing to the ongoing dialogue on this important issue. The evaluators used a five-point Likert scale and a Yes/No system to analyze the living conditions during the fair, including the size of containers, presence of substrate, and environmental enrichment. They also assessed the occurrence of visual abnormal postures and behaviors to gauge the overall level of welfare. To ensure the reliability and consistency of the data and minimize potential bias, each evaluator repeated the rating process three times, with a three-week interval between each session. An average value was then calculated for each aspect. A total of 818 animals were present at the fair, with 688 being reptiles (84.11%) and 130 being amphibians (15.89%). This study revealed that the provision of substrate scored higher for reptiles compared to amphibians, while the size of containers for amphibians received higher ratings than those for reptiles. Visual abnormalities in posture and behavior were more common in reptiles than in amphibians. Display containers for snakes received the lowest ratings and showed more visual abnormalities in posture and behavior, raising concerns about their welfare. Despite the presence of environmental enrichment, the overall level of animal welfare was assessed as being medium/low. Pearson's correlation coefficient indicated good reliability among the evaluators during the assessment process, with most assessments showing values > 0.8. Despite existing regulations for exhibitors, neglect remains prevalent. These findings highlight the potential negative impact of animal exposure at fairs on animal welfare. Display containers were often inadequately sized for the animals, particularly for snakes, chameleons, monitor lizards, and salamanders.

Regulation mechanism of endochondral ossification in Rana zhenhaiensis during metamorphosis based on histomorphology and transcriptome analyses.

Endochondral ossification plays a crucial role in the limb development of amphibians. This study explored the ossification sequence in the hindlimb of Rana zhenhaiensis tadpoles and the correlation between thyroid hormones (THs) and endochondral ossification via histomorphology and transcriptional analyses. Our results suggest that ossification of the femur and tibiofibula was initiated during the period of high THs activity (metamorphosis climax). In addition, the results of differentially expressed gene analyses in the hindlimb and tail showed that systemic factors, transcription factors, and locally secreted factors interacted with each other during the metamorphosis climax to regulate the occurrence of endochondral ossification. These results will enrich the morphological data of anurans and provide scientific reference for the evolutionary history of vertebrates.

Early life exposure to high temperature enhances locomotor performance without alteration in thermal ecology in different populations of Thoropa taophora tadpoles (Anura, Cycloramphidae).

Understanding the intricate relationship between temperature and physiological processes in ectotherm vertebrates is crucial for predicting how these animals respond to environmental changes, including those associated with climate change. This is particularly relevant for the anurans, given their limited capacity for thermoregulation, particularly in larval stages. Herein, we investigated the capacity for thermal acclimatization in Thoropa taophora tadpoles, an endemic species in the Atlantic rainforest of Southeast Brazil, inhabiting distinct thermal environments. These semi-terrestrial tadpoles develop on rocky surfaces with some populations inhabiting exposed regions near the marine coast where temperatures may reach up to 30 °C in sunny conditions, while other populations occupy forested areas near waterfalls that maintain lower temperatures. We aimed to understand the effects of temperature on locomotor performance and on the activity of metabolic enzymes that support performance in tadpoles sampled in four different populations. Moreover, we measured several aspects of thermoregulation including the critical thermal maximum (CTmax), the body temperature of activity (Tb), the preferred temperature (Tpref) and the effectiveness of thermoregulation (E). Despite differences in body size, tadpoles from warmer environments consistently demonstrate higher locomotor performance, with minimal or no acclimatization seen in other variables. Correlations between habitat temperature and biological endpoints underscore the significance of maximum locomotor performance in shaping physiological responses. Our results show how temperature can impact tadpole behavior and performance, without change in many organismal measures of thermal acclimatization, providing insights into potential ecological implications, particularly in the context of climate change.

Generation of genome-wide SNP markers from minimally invasive sampling in endangered animals and applications in species ecology and conservation.

High-density genotyping methods have revolutionized the field of population and conservation genetics in the past decade. To exploit the technological and analytical advances in the field, access to high-quality genetic material is a key component. However, access to such samples in endangered and rare animals is often challenging or even impossible. Here, we used a minimally invasive sampling method (MIS) in the endangered cave salamander Proteus anguinus, the olm, to generate thousands of genetic markers using ddRADseq for population and conservation genomic analyses. Using tail clips and MIS skin swabs taken from the same individual, we investigated genotyping data properties of the two different sampling types. We found that sufficient DNA can be extracted from swab samples to generate up to 200,000 polymorphic SNPs in divergent Proteus lineages. Swab and tissue samples were highly reproducible exhibiting low SNP genotyping error rates. We found that SNPs were most frequently (~50%) located within genic regions, while the rest mapped to mostly flanking regions of repetitive DNA. The vast majority of DNA recovered from swabbing was host DNA. However, a fraction of DNA recovered from swabs contained additional ecological information on the species, including eDNA from the surrounding environment and bacterial skin fauna. Most exogenous DNA recovered from swabs were bacteria (~80%), followed by vertebrates (~20%). Our results demonstrate that MIS can be used to (i) generate tens of thousands of ddRADseq markers for conservation and population genomic analyses and (ii) inform on the species health status and ecology from exogenous DNA.

Staphylococci, Reptiles, Amphibians, and Humans: What Are Their Relations?

Reptiles and amphibians are largely present in many environments, including domestic areas when they are kept as pet animals. They often harbor zoonotic pathogens, which can pose a serious risk of infection for humans, mainly immunocompromised individuals, the elderly, children, and pregnant women. Several studies have been carried out to verify the role of cold-blooded animals in the epidemiology of some bacteria, mainly Salmonella, whereas scarce attention has been focused on these animals as a source of staphylococci. These bacteria are often antimicrobial-resistant and they act as opportunistic pathogens, which can cause relevant infections in humans and animals, both domestic and wild. Asymptomatic reptiles and amphibians often harbor staphylococcal strains, such as Staphylococcus aureus and coagulase-negative Staphylococcus spp.; however, these bacteria have been associated with clinical conditions that usually appear in animals under stress conditions. In all cases, greater attention should also be focused on staphylococci in cold-blooded animals due to their implications in human and veterinary medicine.

Dehydration followed by restraint sustains high circulating corticosterone and improves immunity in toads.

Amphibians are suffering population declines due to a variety of factors such as increased ultraviolet radiation, climate change, habitat loss, pathogens, and pollution, or a combination of these. Such changes are associated with a reduction in the availability of water, exposing these animals to a greater risk of desiccation. In this context, understanding how dehydration can modulate the hypothalamic-pituitary-interrenal axis (HPI) and the immune response is an imperative question to predict how stressors can affect amphibian species. We investigated whether dehydration promotes long-lasting effects on toads' ability to respond to a consecutive stressor (restraint) even if the toads are allowed to rehydrate, as well as its effects on the immune function. We also tested the hypothesis that the toads showing more severe dehydration would exhibit lower responsiveness to restraint challenge, even if the animals were allowed to rehydrate. Individuals of R. ornata were dehydrated mildly and severely. Thereafter, they were submitted to a restraint stress challenge for 1 and 24 h. Our results show that dehydration increased hematocrit and CORT in R. ornata toads. The restraint induced an acute stress response in fully hydrated toads (increased CORT and neutrophil: lymphocyte ratio). Otherwise, restraint in moistened cloth bags allowed full rehydration in previously dehydrated toads and did not induce an additional increase in CORT, but those toads sustained elevated CORT up to 24 h of restraint. Also, these animals showed increased neutrophil: lymphocyte ratio and the phagocytic activity of blood cells, even when they rehydrated during restraint. These results point to a continuous activation of the HPA during dehydration and subsequent restraint, even when they recovered from the dehydration state. Also, acute stressors seem to promote immune cell redistribution and augmentation of immune cellular function in R. ornata toads.

Impact of rising temperature on physiological and biochemical alterations that affect the viability of blood cells in American bullfrog crossbreeds.

The study aimed to examine the impact of increasing environmental temperatures on physiological changes, oxidative stress, nitric oxide production, total antioxidant capacity, and blood cell viability in American bullfrog crossbreeds. Frogs and frog blood cells were exposed to temperature ranges of 25-33 °C and 25-37 °C, respectively. Physiological parameters (body temperature, pulse rate, ventilation rate, and oxygen saturation) and biochemical parameters (total antioxidant power, hydrogen peroxide, malondialdehyde, nitric oxide, and mitochondrial activity) were measured at every 2 °C increment. Results showed that body temperature rose with increased environmental temperature (P < 0.05). Pulse rates at 33 °C were higher than those at 25-31 °C (P < 0.05). Ventilation rates at 31 °C exceeded those at 25 °C and 27 °C (P < 0.05). Oxygen saturation levels remained stable at 25-33 °C (P > 0.05). Total antioxidant power at 25 °C was greater than at 27-37 °C (P < 0.05). Hydrogen peroxide levels at 27 °C were higher compared to 25 °C and 31-37 °C (P < 0.05). Malondialdehyde levels at 25-33 °C were higher than at 35 °C and 37 °C (P < 0.05). Nitric oxide levels at 37 °C were higher than at 25-33 °C (P < 0.05), and at 35 °C were higher than at 25-31 °C (P < 0.05). Blood cell viability at 25-31 °C was higher than at 37 °C (P < 0.05). These results suggest that at an environmental temperature of 33 °C, the frogs' body temperature approached 31 °C or higher, and were likely to be harmful to the frogs. Finally, the environmental temperature that caused frog blood cell death was 37 °C.

Integrative systematic revision of the Montseny brook newt (Calotriton arnoldi), with the description of a new subspecies.

The Montseny brook newt (Calotriton arnoldi), a glacial relict endemic to a small, isolated massif in northeast Spain, is considered the only Critically Endangered urodele in Europe. Its restricted range is divided by a deep valley that acts as an impassable barrier to dispersal, separating two isolated metapopulations (Western and Eastern) that correspond to independent lineages with different evolutionary trajectories, based on genetic and genomic data. Here, we address the ecological differentiation between lineages and discuss its potential effect on the phenotypic distinctness of each lineage. Based on multiple lines of evidence, we formally describe the Western Montseny brook newt as a new subspecies: Calotriton arnoldi laietanus ssp. nov. Finally, our study underscores the importance of considering taxonomic progress in the conservation policies of endangered species, ensuring appropriate management and protection of the newly described taxa.

Limited evidence for genetic differentiation or adaptation in two amphibian species across replicated rural-urban gradients.

Urbanization leads to complex environmental changes and poses multiple challenges to organisms. Amphibians are highly susceptible to the effects of urbanization, with land use conversion, habitat destruction, and degradation ranked as the most significant threats. Consequently, amphibians are declining in urban areas, in both population numbers and abundance, however, the effect of urbanization on population genetic parameters remains unclear. Here, we studied the genomic response to urbanization in two widespread European species, the common toad Bufo bufo (26 localities, 480 individuals), and the smooth newt Lissotriton vulgaris (30 localities, 516 individuals) in three geographic regions: southern and northern Poland and southern Norway. We assessed genome-wide SNP variation using RADseq (ca. 42 and 552 thousand SNPs in toads and newts, respectively) and adaptively relevant major histocompatibility complex (MHC) class I and II genes. The results linked most of the genetic differentiation in both marker types to regional (latitudinal) effects, which also correspond to historical biogeography. Further, we did not find any association between genetic differentiation and level of urbanization at local scales for either species. However, urban smooth newts, but not toads, have lower levels of within-population genome-wide diversity, suggesting higher susceptibility to the negative effects of urbanization. A decreasing level of genetic diversity linked to increasing urbanization was also found for MHC II in smooth newts, while the relationship between MHC class I diversity and urbanization differed between geographic regions. We did not find any effects of urbanization on MHC diversity in the toad populations. Although two genetic environment association analyses of genome-wide data, LFMM and BayPass, revealed numerous (219 in B. bufo and 7040 in L. vulgaris) SNPs statistically associated with urbanization, we found a marked lack of repeatability between geographic regions, suggesting a complex and multifaceted response to natural selection elicited by life in the city.

Path for recovery: an ecological overview of the Jambato Harlequin Toad (Bufonidae: Atelopus ignescens) in its last known locality, Angamarca Valley, Ecuador.

The Jambato Harlequin toad (Atelopus ignescens), a formerly abundant species in the Andes of Ecuador, faced a dramatic population decline in the 1980s, with its last recorded sighting in 1988. The species was considered Extinct by the IUCN until 2016, when a fortuitous discovery of one Jambato by a local boy reignited hope. In this study, we present findings from an investigation conducted in the Angamarca parish, focusing on distribution, abundance, habitat preferences, ecology, disease susceptibility, and dietary habits of the species. In one year we identified 71 individuals at different stages of development in various habitats, with a significant presence in agricultural mosaic areas and locations near water sources used for crop irrigation, demonstrating the persistence of the species in a complex landscape, with considerable human intervention. The dietary analysis based on fecal samples indicated a diverse prey selection, primarily comprising arthropods such as Acari, Coleoptera, and ants. Amphibian declines have been associated with diseases and climate change; notably, our study confirmed the presence of the pathogen Batrachochytrium dendrobatidis (Bd), but, surprisingly, none of the infected Jambatos displayed visible signs of illness. When analyzing climatic patterns, we found that there are climatic differences between historical localities and Angamarca; the temporal analysis also exposes a generalized warming trend. Finally, in collaboration with the local community, we developed a series of management recommendations for terrestrial and aquatic environments occupied by the Jambato.

eDNA Metabarcoding Reveals the Species-Area Relationship of Amphibians on the Zhoushan Archipelago.

The species-area relationship is important for understanding species diversity patterns at spatial scales, but few studies have examined the relationship using environmental DNA (eDNA) techniques. We investigated amphibian diversity on 21 islands of the Zhoushan Archipelago and nearby mainland areas in China using the combination of eDNA metabarcoding and the traditional line transect method (TLTM) and identified the species-area relationship for amphibians on the islands. The mean detection probability of eDNA is 0.54, while the mean detection probability of TLTM is 0.24. The eDNA metabarcoding detected eight amphibian species on the islands and nine species in the mainland areas, compared with seven species on the islands and nine species in the mainland areas that were identified by TLTM. Amphibian richness on the islands increased with island area and habitat diversity. The species-area relationship for amphibians in the archipelago was formulated as the power function (S = 0.47A0.21) or exponential function (S = 2.59 + 2.41 (logA)). Our results suggested that eDNA metabarcoding is more sensitive for the detection of amphibian species. The combined use of eDNA metabarcoding and the traditional line transect method may optimize the survey results for amphibians.

An Annotated Checklist of Monogeneans (Platyhelminthes, Monogenea) from Aquatic Vertebrates in Peru: A Review of Diversity, Hosts and Geographical Distribution.

Monogeneans are flatworm parasites that infest fish gills primarily but can also infect various other vertebrates, including amphibians, aquatic reptiles, mammals like hippos, and occasionally invertebrates like copepods, isopods, and cephalopods. Despite their remarkable diversity, our knowledge of monogenean parasites in Peru remains significantly limited, resulting in substantial gaps in our comprehension of their taxonomic identities, host associations, and geographic distribution. To address these knowledge deficits, we present an extensively curated checklist of monogeneans associated with aquatic vertebrates in Peru. This comprehensive compilation is derived from meticulous literature surveys, the examination of specimens deposited in both international and national collections, and the inclusion of additional freshly collected specimens. The checklist offers a thorough repository of data encompassing the diversity, host associations, and geographical distribution of these parasites. Taxonomic discrepancies are addressed through a critical review of the existing literature, supplemented by the direct examination of specimens, including type or voucher specimens, deposited within scientific collections. Additionally, we provide data on the DNA sequences of individual taxa. The compiled list comprises records of 358 monogenean species, including 270 valid species and 88 taxa identified at the family or generic level, all reported across 145 host species in Peru. Predominantly, these parasitic species exhibit associations within fish, with 335 infecting teleosts and 20 affecting chondrichthyans. Three monogenean species have been documented as infecting amphibians, namely Mesopolystoma samiriensis, Polistoma sp. and Wetapolystoma almae. Among the monogeneans reported, 141 were found in marine environments and 214 in freshwater environments. The most diverse families were Dactylogyridae and Diplectanidae, comprising 217 and 24 species, respectively. The hosts that harbored the highest number of monogeneans were Pygocentrus nattereri (with 23 species), followed by Stellifer minor (13 spp.) and Triportheus angulatus (11 spp.). We detected many species that do not have any material deposited in a scientific collection due to the loss or deactivation of the collection. These findings represent only a fraction of the potential diversity, considering the wide variety of aquatic vertebrate hosts inhabiting the tropical and subtropical regions of Peru.