Amphibian Species Research Articles

Endemic amphibians of the Cerrado and Caatinga: species richness, geographic range and conservation

The Cerrado and the Caatinga have few formally protected areas and encompass a high diversity of amphibians, which is still relatively unknown. In this study we present the list of amphibian species endemic to the Cerrado or Caatinga and their geographic range, highlighting current conservation status. We obtained the species list from the compilation of scientific publications (up to July 2022). We obtained the occurrence records from the databases SpeciesLink, GBIF, Portal da Biodiversidade and literature. We classified the species in the categories of threat according to the Brazilian list of threatened species and the global list of threatened species. We compiled 2,659 occurrences of amphibians, 1,335 from the SpeciesLink platform, 414 from GBIF, 371 from the Biodiversity Portal and 539 obtained from the literature. We recorded 100 endemic species, 82 from the Cerrado and 18 from the Caatinga. The Cerrado has three species Vulnerable (VU) (Boana buriti, Bokermannohyla napolii and Scinax pinimus), one Critically Endangered (CR) (Proceratophrys moratoi), 14 species Data Deficient (DD), 46 in the Least Concern (LC) category and 18 Not Evaluated (NE). The Caatinga has three species CR (Adelophryne maranguapensis, Proceratophrys ararype, Rhinella casconi), three DD, five LC and seven NE. Threatened, DD and NE species have few records within the protected areas of Cerrado and Caatinga. We consider that the Cerrado and Caatinga present a rich diversity of endemic amphibians, which have a geographic range relatively reduced, especially the threatened ones, DD and NE. Distribution and taxonomy data are essential, because the lack can hinder the assessment of conservation status, since threatened species, DD, NE, including LC, may be undervalued and at risk. Keywords: amphibia, distribution, endemism, species list, species protection

Contrasting effects of ammonium nitrate on tadpole survival, growth and behavior in two common anuran species

Extensive fertilizer use often leads to an accumulation of chemicals in waterbodies that are in close proximity to agricultural fields. One such commonly used fertilizer, ammonium nitrate, has been shown to negatively affect anuran survival and behavior. We performed experiments on tadpoles of two amphibian species, the Agile frog (Rana dalmatina) and the Common toad (Bufo bufo), to test the effects of different concentrations (0 mg/L, 50 mg/L, 100 mg/L and 200 mg/L) of ammonium nitrate on survival, growth and movement activity. We also examined whether sensitivity to predator cues might be altered in the presence of ammonium nitrate. Finally, we tested for the capacity of the two species to exhibit avoidance behavior. Overall, we found that the Agile frog is more sensitive to ammonium nitrate than the Common toad in terms of survival, growth and movement activity. Sensitivity to predator cues was not affected by the fertilizer for the Agile frog, but was shown to alter the behavior of Common toads, whose movement activity did not decrease in the presence of predator cues. Neither species showed an avoidance behavior. Our results suggest that ammonium nitrate can have significant effects on Agile frog tadpoles through direct toxicological effects on survival and growth, but also through behavioral effects. For Common toads those effects were less obvious, but were nonetheless shown in terms of growth, activity and predator detection.

Amphibian conservation status in Brazil: Spatial patterns, threats, and challenges

Extinction risk assessments are valuable conservation tools, especially for sensitive groups like amphibians, which are facing escalating threats from habitat loss and degradation. In this scenario, Brazil stands out as a pivotal player in amphibian conservation due to its exceptional amphibian diversity. Between 2010 and 2014, 973 amphibian species were assessed in Brazil. More recently, from 2017 to 2018, this number increased to 1096. In this study, we (1) compare the extinction risk status of Brazilian amphibians between the 1st and 2nd assessments; (2) evaluate the reasons for changes in status; (3) identify the main threats and research recommendations; and (4) analyze the spatial patterns of threatened species in relation to protected areas and ecoregions in Brazil. Our findings reveal an increase in the number of threatened species from the first to the second assessment. The categories of Least Concern and Critically Endangered experienced the most significant increases, with 169 and 16 more species, respectively, while the count of Data Deficient species dropped by 71. The primary factor contributing to these changes is the availability of new, more precise information, affecting 134 species, or 85.4% of the cases. Threatened species are predominantly clustered within the Atlantic Forest biome, itself the most endangered biome in Brazil, hosting 46 species, or 78% of the total. Notably, one-third of threatened species in Brazil do not inhabit protected areas and thus require immediate conservation attention. While substantial progress has been made in understanding Brazilian amphibians in recent years, an urgent need persists for fundamental research. This research should encompass taxonomy, natural history, and the evaluation of threat impacts, providing a robust foundation for more effective conservation efforts concerning this group.

Interspecific cytogenomic comparison reveals a potential chromosomal centromeric marker in Proceratophrys frog species.

Among the repetitive elements, satellite DNA (SatDNA) emerges as extensive arrays of highly similar tandemly repeated units, spanning megabases in length. Given that the satDNA PboSat01-176, previously characterized in P. boiei, prompted our interest for having a high abundance in P. boiei and potential for centromeric satellite, here, we employed various approaches, including low coverage genome sequencing, followed by computational analysis and chromosomal localization techniques in four Proceratophrys species and, investigating the genomic presence and sharing, as well as its potential for chromosomal centromere marker in Proceratophrys frog species. Our findings demonstrate that PboSat01-176 exhibits high abundance across all four Proceratophrys species, displaying distinct characteristics that establish it as the predominant repetitive DNA element in these species. The satellite DNA is prominently clustered in the peri/centromeric region of the chromosomes, particularly in the heterochromatic regions. The widespread presence of PboSat01-176 in closely related Proceratophrys species reinforces the validity of the library hypothesis for repetitive sequences. Thus, this study highlighted the utility of the satDNA family PboSat01-176 as a reliable centromeric marker in Proceratophrys species, with potential to be applied in other species of anuran amphibians. The observed sharing and maintenance of this sequence within the genus suggest possibilities for future research, particularly through expanded sampling to elucidate parameters that underlie the library hypothesis and the evolutionary dynamics of satDNA sequences.

Unnecessary splitting of genus-level clades reduces taxonomic stability in amphibians

Abstract Although the differentiation of clades at the species level is usually based on a justifiable and testable conceptual framework, the demarcation of supraspecific boundaries is less objective and often subject to differences of opinion. The increased availability of large-scale phylogenies has in part promulgated a practice of what we consider excessively splitting clades at the “genus” level. Many of these new genus-level splits are predicated on untenable supporting evidence (e.g., weakly supported phylogenies and purportedly “diagnostic” but actually variable, non-exclusive, or otherwise problematic opposing character state differences) without careful consideration of the effects on downstream applications. As case studies, we critically evaluate several recent examples of splitting established monophyletic genera in four amphibian families that resulted in the creation/elevation of 20 genus-level names (Dicroglossidae: Phrynoglossus, Oreobatrachus, Frethia split from Occidozyga; Microhylidae: Nanohyla split from Microhyla; Ranidae: Abavorana, Amnirana, Chalcorana, Humerana, Hydrophylax, Indosylvirana, Papurana, Pulchrana, Sylvirana split from Hylarana; Rhacophoridae: Tamixalus, Vampyrius, Leptomantis, Zhangixalus split from Rhacophorus, Rohanixalus split from Feihyla, Orixalus split from Gracixalus, and Taruga split from Polypedates), and also address the taxonomic status of the monotypic genus Pterorana relative to Hylarana. We reassess the original claims of diagnosability and justifications for splitting and argue that in many cases, the generic splitting of clades is not only unnecessary but also destabilizes amphibian taxonomy, leading to a host of downstream issues that affect categories of the user community (stakeholders such as taxonomists, conservationists, evolutionary biologists, biogeographers, museum curators, educators, and the lay public). As an alternative, we advocate for the use of the subgenus rank in some cases, which can be implemented to establish informative partitions for future research without compromising on information content, while avoiding gratuitous (and often transient) large-scale binomial (genus-species couplet) rearrangements. We encourage taxonomists to consider the actual needs and interests of the larger non-taxonomic end-user community who fund the majority of taxonomic research, and who require a system that remains reasonably stable and is relatively intuitive, without the need for inaccessible laboratory equipment or advanced technical scientific knowledge to identify amphibian species to the genus level.

Evaluating potential developmental toxicity of perfluoroalkyl and polyfluoroalkyl substances in Xenopus laevis embryos and larvae.

As part of the US Environmental Protection Agency's perfluoroalkyl and polyfluoroalkyl substances (PFAS) Action Plan, the agency is committed to increasing our understanding of the potential ecological effects of PFAS. The objective of these studies was to examine the developmental toxicity of PFAS using the laboratory model amphibian species Xenopus laevis. We had two primary aims: (1) to understand the developmental toxicity of a structurally diverse set of PFAS compounds in developing embryos and (2) to characterize the potential impacts of perfluorooctanesulfonic acid (PFOS), perfluorohexanesulfonic acid (PFHxS), perfluorooctanoic acid (PFOA), and hexafluoropropylene oxide-dimer acid (HFPO-DA a.k.a. GenX), on growth and thyroid hormone-controlled metamorphosis. We employed a combination of static renewal and flow-through exposure designs. Embryos were exposed to 17 structurally diverse PFAS starting at the midblastula stage through the completion of organogenesis (96 h). To investigate impacts on PFOS, PFOA, PFHxS, and HFPO-DA on development and metamorphosis, larvae were exposed from premetamorphosis (Nieuwkoop Faber stage 51 or 54) through pro metamorphosis. Of the PFAS tested in embryos, only 1H,1H,10H,10H-perfluorodecane-1,10-diol (FC10-diol) and perfluorohexanesulfonamide (FHxSA) exposure resulted in clear concentration-dependent developmental toxicity. For both of these PFAS, a significant increase in mortality was observed at 2.5 and 5mg/L. For FC10-diol, 100% of the surviving embryos were malformed at 1.25 and 2.5mg/L, while for FHxSA, a significant increase in malformations (100%) was observed at 2.5 and 5mg/L. Developmental stage achieved was the most sensitive endpoint with significant effects observed at 1.25 and 0.625 mg/L for FC10-diol and FHxSA, respectively. In larval studies, we observed impacts on growth following exposure to PFHxS and PFOS at concentrations of 100 and 2.5mg/L, respectively, while no impacts were observed in larvae when exposed to PFOA and HFPO-DA at concentration of 100 mg/L. Further, we did not observe impacts on thyroid endpoints in exposed larvae. These experiments have broadened our understanding of the impact of PFAS on anuran development.

Фенология начала нерестовых миграций бесхвостых амфибий (Anura, Amphibia) в долинах рек Саратовского Правобережья

The intra-century dynamics of the spawning migrations start of the three most numerous species of anuran amphibians (Pelobates vespertinus, Bombina bombina and Pelophylax ridibundus) was analyzed. Local populations of the above anurans in the valley of the middle course of the Medveditsa River (Don Basin, Saratov region) were the object of the study. From 1892 to 2023 the phenological norm of this phase of the annual cycle shifted to earlier dates by an average of 7 days for each species in a climate warming. By 2023, the henological norm of the spawning migration start of P. vespertinus, B. bombina and P. ridibundus was April 14, April 21, and April 22, respectively. In addition, the change rate of this phenological parameter in the model species was similar as well, about 8 days/100 years. The absolute value of the phenological norm change, as well as the rate of its transformation, is comparable among the three species of anuran amphibians, despite any differences in their thermobiological characteristics. The early snow clearence in the Northern Lower Volga region may be one of the reasons for shifting the start dates of amphibian spawning migrations to earlier dates. The continued phenological changes in the spring phase of the annual cycle require continued monitoring of this process. This will allow forming scientifically based forecasts of local population dynamics in the future. Possible consequences of this transformation of the spring phenology of anuran amphibians are increased probability of metamorph survival in low water availability in the floodplain, and spawning migration development in the false spring scenario and susceptibility to return cold weather. This is discussed in relation to the changes identified.

At home in Jiangsu: Environmental niche modeling and new records for five species of amphibian and reptile in Jiangsu, China

Environmental niche models are useful tools for generating hypotheses for the distribution of species and informing conservation planning, especially at the edge of species’ ranges and for those with limited data. Here we report on the recent documentation of four species of amphibian (Hylarana latouchii, Odorrana tianmuii, Polypedates braueri, and Zhangixalus dennysi) and one reptile (Protobothrops mucrosquamatus) with few or no previous geolocated records from Jiangsu, China. We combined our opportunistic field sampling data from Jiangsu, which is at the edge of each of these species’ ranges, with publicly available occurrence records and climatic data to generate environmental niche models for these five species using Maxent. All models showed good model performance with AUC values ranging from 0.899 to 0.983. Additional potentially suitable areas within southern Jiangsu were predicted for the four amphibian species, although the significant anthropogenic habitat modifications in the province may limit their contemporary distributions. For all five species, the climatic variable that contributed most to the model was the precipitation of the driest month (Bio 14), indicating they are limited by moisture availability. Our study adds new information about the climatic preferences of these five species and highlights the value of complementing environmental niche modeling with field surveys for robust inferences and conservation planning, particularly at the edge of species’ ranges.

Immunohistochemical Characterization of Langerhans Cells in the Skin of Three Amphibian Species.

The amphibian taxon includes three orders that present different morphological characteristics: Anura, Caudata, and Apoda. Their skin has a crucial role: it acts as an immune organ constituting a physical, chemical, immunological, and microbiological barrier to pathogen insult and conducts essential physiological processes. Amphibians have developed specialized features to protect the vulnerable skin barrier, including a glandular network beneath the skin surface that can produce antimicrobial and toxic substances, thus contributing to the defense against pathogens and predators. This study aims to characterize Langerhans cells in the skin of Lithobates catesbeianus (order: Anura; Shaw, 1802), Amphiuma means (order: Caudata; Garden, 1821), and Typhlonectes natans (order: Apoda; Fischer, 1880) with the following antibodies: Langerin/CD207 (c-type lectin), Major Histocompatibility Complex (MHC)II, and Toll-like receptor (TLR)2 (expressed by different types of DCs). Our results showed Langerhans cells positive for Langerin CD/207 in the epidermis of the three species; moreover, some antigen-presenting cells (APCs) in the connective tissue expressed TLR2 and MHCII. The distribution of the Langerhans cells is very similar in the three amphibians examined, despite their different habitats. A greater knowledge of the amphibian immune system could be useful to better understand the phylogeny of vertebrates and to safeguard amphibians from population declines. Furthermore, the similarities between amphibians' and human skin concerning immunological features may be useful in both biology and translational medicine.

The importance of quantifying fitness-determining traits throughout life to assess the application of reproductive technologies for amphibian species recovery

The application of reproductive technologies to amphibian conservation breeding programs is gaining momentum and the field is poised to contribute significantly toward amphibian species recovery. We briefly discuss the opportunities for reproductive technologies to enhance conservation breeding outcomes, including their potential to enhance the genetic management, and in turn, the fitness of threatened species. Despite this potential, an important consideration that is not yet well understood is the degree to which specific reproductive technologies might influence (either enhance, or in some instances potentially decrease) individual fitness and lead to shifts in population viability. The development of a standardised approach to monitoring offspring throughout life-stages to detect changes to morphology, behaviour, physiology, survivorship, and developmental trajectories is essential. The primary focus of this review is to provide a ‘best-practise’ framework for quantifying key fitness determining traits expected to contribute to the fitness of individuals and long-term viability of populations, which will ultimately allow us to progress the field of amphibian reproductive technologies and assess the impact of protocol refinement.

Ranavirus and helminth parasite co-infection in invasive American bullfrogs in the Atlantic forest, Brazil

Emerging infectious diseases threaten amphibian species across the globe. In Brazil, the American bullfrog (Aquarana catesbeiana) is a highly invasive species that can potentially transmit parasites and pathogens to native amphibians. This is the first assessment of co-infection of Ranavirus and helminth macroparasites in invasive populations of bullfrogs in South America. We collected, measured, and euthanized 65 specimens of A. catesbeiana sampled from 9 sites across three states of Brazil in the Atlantic Forest biome. We collected and identified helminth macroparasites and sampled host liver tissue to test for the presence and load of Ranavirus with quantitative PCR. We documented patterns of prevalence, parasite load, and co-infection with generalized linear mixed models, generalized logistic regressions, and randomization tests. Most individual bullfrogs did not exhibit clinical signs of infection, but the overall Ranavirus prevalence was 27% (95% confidence interval, [CI 17–38]). Bullfrogs were infected with helminth macroparasites from 5 taxa. Co-infection of helminth macroparasites and Ranavirus was also common (21% CI [12–31]). Bullfrog size was positively correlated with total macroparasite abundance and richness, and the best-fitting model included a significant interaction between bullfrog size and Ranavirus infection status. We observed a negative correlation between Ranavirus viral load and nematode abundance (slope = −0.22, P = 0.03). Invasive bullfrogs (A. catesbeiana) in Brazil were frequently infected with both Ranavirus and helminth macroparasites, so adult bullfrogs could serve as reservoir hosts for both pathogens and parasites. However, many macroparasites collected were encysted and not developing. Coinfection patterns suggest a potential interaction between Ranavirus and macroparasites because helminth abundance increased with bullfrog size but was lower in Ranavirus infected individuals. Future studies of bullfrogs in the Atlantic Forest should investigate their potential role in pathogen and parasite transmission to native anurans.

An extensive database on the traits and occurrences of amphibian species in Turkey

Amphibians are the most endangered taxa among vertebrates, and they face many threats during their complex life cycles. The species’ life history traits and occurrence database help understand species responses against ecological factors. Consequently, the species-level-trait database has gained more prominence in recent years as a useful tool for understanding the dimensions of communities, assembly processes of communities, and conserving biodiversity at the ecosystem level against environmental changes. However, in Turkey, there are deficiencies in the knowledge of the ecological traits of amphibians compared to other vertebrate taxa, as most studies have focused on their distribution or taxonomic status. Consequently, there is a need to create such a database for future research on all known extant amphibians in Turkey. We compiled a species-level data set of species traits and occurrences for all amphibians in Turkey using 436 literature sources. We completed 36 trait categories with 5611 occurrence data for 37 amphibian species in Turkey. This study provides an open, useful, and comprehensive database for macroecological and conservation studies on amphibians in Turkey.

Diversity and Molecular Evolution of Antimicrobial Peptides in Caecilian Amphibians.

Antimicrobial peptides (AMPs) are key molecules in the innate immune defence of vertebrates with rapid action, broad antimicrobial spectrum, and ability to evade pathogen resistance mechanisms. To date, amphibians are the major group of vertebrates from which most AMPs have been characterised, but most studies have focused on the bioactive skin secretions of anurans (frogs and toads). In this study, we have analysed the complete genomes and/or transcriptomes of eight species of caecilian amphibians (order Gymnophiona) and characterised the diversity, molecular evolution, and antimicrobial potential of the AMP repertoire of this order of amphibians. We have identified 477 candidate AMPs within the studied caecilian genome and transcriptome datasets. These candidates are grouped into 29 AMP families, with four corresponding to peptides primarily exhibiting antimicrobial activity and 25 potentially serving as AMPs in a secondary function, either in their entirety or after cleavage. In silico prediction methods were used to identify 62 of those AMPs as peptides with promising antimicrobial activity potential. Signatures of directional selection were detected for five candidate AMPs, which may indicate adaptation to the different selective pressures imposed by evolutionary arms races with specific pathogens. These findings provide encouraging support for the expectation that caecilians, being one of the least-studied groups of vertebrates, and with ~300 million years of separate evolution, are an underexplored resource of great pharmaceutical potential that could help to contest antibiotic resistance and contribute to biomedical advance.

Variation in amphibian maturation rates influences population vulnerability to disease‐induced declines

AbstractUnderstanding factors that influence population‐level responses to emerging threats in declining species is crucial for informed conservation action. In amphibian species impacted by the chytrid fungus (Batrachochytrium dendrobatidis), a pathogen that has caused amphibian declines globally, a commonly reported pattern is that more severe population declines tend to occur at higher elevations. Previous research has suggested that this pattern could be driven by reduced environmental suitability for chytrid fungus at lower elevations. However, delayed amphibian maturation, which is common in cold, high elevation populations, could also increase vulnerability to population decline. Here, we tackle this key knowledge gap, focusing on the critically endangered corroboree frogs (Pseudophryne corroboree and P. pengilleyi), which have experienced a pattern of extirpation at higher elevations, with remnant populations persisting at lower elevations. First, we quantify the age structure of two extant low elevation P. pengilleyi populations and museum specimens (both species) collected before the emergence of chytrid fungus in Australia. Male age to maturation varied from 1 to 3 years, with the extant population with higher chytrid prevalence displaying severe age structure truncation. Second, we use population simulations to calculate elasticity values under a range of scenarios with varying ages to maturation and chytrid‐associated mortality. When the population growth rate was fixed at 1, adult survival became increasingly important as age to maturation increases, particularly under a scenario of high chytrid‐associated mortality. Our simulation results indicate that delayed maturation could be a previously underappreciated factor associated with an increased risk of amphibian population decline and that earlier maturation could contribute to population persistence. Our study highlights the importance of examining variation in life history traits to better understand population‐level responses to novel threats and guide the development of appropriate conservation actions.

Limited contribution of photoenzymatic DNA repair in mitigating carry-over effects from larval UVB exposure: Implications for frog recruitment

Solar ultraviolet-B (UVB) radiation has increased due to stratospheric ozone depletion, climate and ecosystem changes and is a driver of amphibian population declines. Photoenzymatic repair (PER) is a critical mechanism for limiting UVB lethality in amphibian larvae. However, the link between PER and the UVB-induced effects remains understudied through long-term investigations in vivo. Here, we assessed how larval PER determines the lethal and sublethal effects induced by environmentally relevant acute UVB exposure until the juvenile phase in the Neotropical frog Odontophrynus americanus. We conducted laboratory-based controlled experiments in which tadpoles were or were not exposed to UVB and subsequently were exposed to light (for PER activation) or dark treatments. Results showed that the rates of mortality and apoptosis observed in post-UVB dark treatment are effectively limited in post-UVB light treatment, indicating PER (and not dark repair, i.e. nucleotide excision repair) is critical to limit the immediate genotoxic impact of UVB-induced pyrimidine dimers. Nonetheless, even tadpoles that survived UVB exposure using PER showed sublethal complications that extended to the juvenile phase. Tadpole responses included alterations in morphology, chromosomal instability, increased skin susceptibility to fungal proliferation, as well as increased generation of reactive oxygen species. The short-term effects were carried over to later stages of life because metamorphosis time increased and juveniles were smaller. No body abnormalities were visualized in tadpoles, metamorphs, and juveniles, suggesting that O. americanus is UVB-resistant concerning these responses. This study reveals that even frog species equipped with an effective PER are not immune to carry-over effects from early UVB exposure, which are of great ecological relevance as late metamorphosis and smaller juveniles may impact individual performance and adult recruitment to breeding. Future ecological risk assessments and conservation and management efforts for amphibian species should exercise caution when linking PER effectiveness to UVB resistance.

Identifying Strategies for Effective Biodiversity Preservation and Species Status of Chilean Amphibians.

Resources are limited in global biodiversity conservation efforts, which emphasizes the significance of setting conservation priorities. Using standardized criteria, we evaluated 58 amphibian species in Chile to determine their conservation priority (CP). Species with insufficient historical data had their values marked as missing. With a median value of p = 1.67, the results demonstrated CP values ranging from p = 0.48 to p = 3.0, classifying species into priority and non-priority groups. Four levels were established for the priority categories: no priority, low priority, medium priority, and high priority. Additionally, the Telmatobiidae and Alsodidae families were identified as two more priority families. Notably, the species with the highest priority were found to be T. halli, T. fronteriensis, T. philippii, T. chusmisensis, A. pehuenche, and Alsodes tumultuosus, where T. philippii and T. fronteriensis have equal priority for conservation at the national level according to the conservation priority analysis. Eight priority families-the Alsodidae, Batrachylidae, Bufonidae, Ceratophryidae, Leptodactylidae, Rhinodermatidae, and Telmatobiidae-were determined, and 14 species-or 24% of the species examined-need further study. Based on the conservation priority analysis, the species T. fronteriensis and T. philippii share the highest priority for conservation at the national level (p = 2.50). With 70% of the amphibians under study being threatened mainly by habitat loss, pollution, and emerging diseases, the creation of conservation categories made the threat assessment process easier. Due to a lack of information on geographic distribution and abundance, quantitatively classifying amphibians in Chile remains difficult. The analysis of conservation priorities and potential extinction threats informs appropriate management strategies.

Improvement of Soil Water Assessment Tool (SWAT) wetland module for modelling of ephemeral pond hydrology

AbstractEphemeral ponds (EPs) are seasonally flooded isolated wetlands that provide a variety of hydroecological benefits, including the provision of breeding habitat for several amphibian and invertebrate species. However, the lack of their explicit representation in hydrological models limits a comprehensive understanding of their interaction with surrounding landscapes and their vulnerability in the context of human interventions and climate change. The purpose of this research was to improve the isolated wetland module of the Soil Water Assessment Tool (SWAT) to better represent EP hydrology. The changes include (1) representation of groundwater and hypodermic flow as the only inflows from the pond drainage surface, due to the intermittent and negligible presence of inflow from surface runoff in forested ponds, (2) revision of how evapotranspiration within EPs is represented and (3) implementation of distinct volume‐area‐depth relationships for ponds based on their geometrical shape. The accuracy of these improvements was assessed against that of a previous isolated wetland formulation in replicating water depth observations of 10 EPs of a portion of the Kenauk forest (68 km2) in the Canadian Shield of the Outaouais region (Québec, Canada). The comparison results show that the revised SWAT model presented here significantly improves the distinct filling and drying water cycle of EPs (average root mean square error of 0.1 m of the revised model vs. 0.23 m for the original model). Besides, the new module allowed to identify that hypodermic flow, evapotranspiration and seepage to the underlying soil are the main EP source and sinks. The new module also allowed to explicitly quantify the differences in filling/drying pattern of the EPs of the Kenauk forest and unlike the original model structure, the new module was able to closely replicate the interannual variation of spring and annual hydroperiod duration.

Mitogenomics reveals extremely low genetic diversity in the endangered Jilin clawed salamander: Implications for its conservation.

The Jilin clawed salamander (Onychodactylus zhangyapingi) is an endemic, endangered, and level-two protected amphibian species of China. In the context of serious threats to amphibians worldwide, conservation studies of this endangered species are urgently needed. In this study, mitogenomic conservation genetics and species distribution modeling analyses were performed for O. zhangyapingi. Sixty-three samples were collected from nine different locations, and the complete mitochondrial genome was sequenced. Population genetic analyses revealed that O. zhangyapingi exhibits only one genetic structure with extremely low nucleotide diversity. Late Pleistocene climate cooling may have led to a reduction in effective population size and extremely low mitogenomic nucleotide diversity in this salamander, and the subsequent temperature increase (~20 kya to present) provided the opportunity for rapid population growth. The continuous highly suitable region for O. zhangyapingi is only approximately 3000 km2 on the southeastern boundary of Jilin Province, China. Fortunately, there are three large forested national nature reserves within the distribution of O. zhangyapingi that can effectively protect endangered species. Our findings suggest that O. zhangyapingi is a vulnerable species with a narrow distribution and extremely low genetic diversity, and we should pay more attention to the conservation management of this species.

A portable Raspberry Pi-based camera set-up to record behaviours of frogs and other small animals under artificial or natural shelters in remote locations.

We describe a Raspberry Pi-based camera system that is portable, robust and weatherproof, with a close-up focus (2.5 cm). We show that this camera system can be used in remote locations with high rainfall and humidity. The camera has an Infrared LED light to film in dark places and can continuously record up to 21 days (504 h). We also describe how to make concrete artificial shelters to mount the camera in. One of the great strengths of this shelter/camera set-up is that the animals choose to take up residence and can then be filmed for extended periods with no disturbance. Furthermore, we give examples of how shelters and cameras could be used to film a range of behaviours in not only many small cryptic amphibian species but also other small vertebrates and invertebrates globally.

Microplastics and TiO2 nanoparticles mixture as an emerging threat to amphibians: A case study on bullfrog embryos

Emerging contaminants can act as contributing factors to the decline of amphibian populations worldwide. Recently, scientists have drawn attention to the potential ecotoxicity of microplastics and nanomaterials in amphibians, however, their possible effects on embryonic developmental stages are still absent. Thus, the present study analyzed the developmental toxicity of environmentally relevant concentrations of polyethylene microplastics (PE MPs; 60 mg/L) and titanium dioxide nanoparticles (TiO2 NPs; 10 μg/L), isolated or in combination (Mix group) on bullfrog embryos, Aquarana catesbeiana, adapting the Frog Embryo Teratogenesis Assay (FETAX, 96h). Allied to the FETAX protocol, we also analyzed the heart rate and morphometric data. The exposure reduced the survival and hatching rates in groups exposed to TiO2 NPs, and to a lesser extent, also affected the Mix group. TiO2 NPs possibly interacted with the hatching enzymes of the embryos, preventing hatching, and reducing their survival. The reduced effects in the Mix group are due to the agglomeration of both toxicants, making the NPs less available for the embryos. PE MPs got attached to the gelatinous capsule of the chorion (confirmed by fluorescence microscopy), which protected the embryos from eventual direct effects of the microplastics on the hatching and survival rates. Although there were no cardiotoxic effects nor morphometric alterations, there was a significant increase in abdominal edemas in the hatched embryos of the PE MPs group, which indicates that osmoregulation might have been affected by the attachment of the microplastics on the embryos’ gelatinous capsule. This study presents the first evidence of developmental toxicity of environmental mixtures of microplastics and nanoparticles on amphibians and reinforces the need for more studies with other amphibian species, especially neotropical specimens that could present bigger sensibility. Our study also highlighted several features of the FETAX protocol as useful tools to evaluate the embryotoxicity of several pollutants on amphibians.