Rana Temporaria Research Articles

Greater Glasgow pond amphibian surveys 2022

In 2019, Clyde Amphibian and Reptile Group (CARG) decided to organise a spring 2020 common frogspawn (Rana temporaria) survey of the Greater Glasgow area, Scotland. The COVID-19 lockdown postponed this project until 2022. The 2022 survey area was spread across four council districts: Glasgow City, South Lanarkshire, Renfrewshire and East Renfrewshire. The primary aim of the surveys was to collect records of amphibian breeding throughout the Greater Glasgow Area, with secondary aims of: (1) revisiting the ponds to see if the ponds supported the spawn through to tadpole and froglet stages; (2) assessing the condition of the ponds for amphibians; (3) identifying areas for habitat enhancement work; and (4) collecting data on breeding common toads (Bufo bufo) and newts (Lissotriton, Ichthyosaura and Triturus spp.). A total of 45 volunteers participated in the surveys. Surveys were completed between March and May 2022, with a primary focus between March and April in order to capture frog spawning data. Surveyors were asked to record amphibian population data, and habitat data in order to estimate Habitat Suitability Indices. In total 162 ponds were surveyed, with evidence of breeding common frogs found in 66.1% of ponds with 11,478 frogspawn clumps counted on the first set of surveys. Only 12.2% of ponds were found to contain common toads, and small newts (Lissotriton spp.) were identified in only 7.4% of ponds. In relation to the quality of breeding habitat, 24.5% of ponds were assessed as drying out annually, which significantly reduces the suitability of these ponds for breeding. This figure is predicted to increase with anthropogenic climate change reducing the number of breeding ponds available to amphibians in the absence of intervention. Furthermore, the majority of the ponds surveyed were assessed as offering poor or below average suitability for great crested newts (Triturus cristatus) and nearly half of the ponds surveyed were found to have poor or bad water quality, which can increase stress levels for larval amphibians and may adversely affect mortality levels. However, as the analysis of water quality was subjective, these findings should be treated with caution.

An overview of the Scottish National Adder Survey 2022-24

A study was commissioned by NatureScot to analyse changes in the status of the adder (Vipera berus) and other native reptiles at 16 sites across mainland Scotland, the Isle of Skye and the Isle of Arran over a 30-year period from 1992-93 to 2022-24. In 2022-24, volunteer surveyors revisited sites that were previously surveyed in 1992-93 to identify changes in occupancy. Visual surveys recorded adder, common lizard (Zootoca vivipara), slow-worm (Anguis fragilis), common frog (Rana temporaria), common toad (Bufo bufo), smooth newt (Lissotriton vulgaris), and palmate newt (Lissotriton helveticus). Adders were observed in comparable or even greater numbers at eight sites (seven of the original sites surveyed in 1992-93), but an apparent loss of adders at three of the original sites, and major declines at a previously favourable site on the Scottish Borders are cause for concern. Of the other reptiles, common lizards were the most widespread, recorded at 12 sites; and slow-worms were least often recorded, in low numbers at only four sites, though this could be expected in visual surveys for this cryptic species. This publication reviews the factors most likely to be affecting adders in Scotland, particularly the loss of suitable habitat, the likely historic impact of persecution and the potential effects of disturbance by recreational site users. It highlights the importance of sympathetic habitat management, with a particular focus on maintaining favourable micro-habitats for reptiles.

Temperature and land use change are associated with Rana temporaria reproductive success and phenology.

Chemical pollution, land cover change, and climate change have all been established as important drivers of amphibian reproductive success and phenology. However, little is known about the relative impacts of these anthropogenic stressors, nor how they may interact to alter amphibian population dynamics. Addressing this gap in our knowledge is important, as it allows us to identify and prioritise the most needed conservation actions. Here, we use long-term datasets to investigate landscape-scale drivers of variation in the reproductive success and phenology of UK Common frog (Rana temporaria) populations. Consistent with predictions, we found that increasing mean temperatures resulted in earlier initialisation of spawning, and earlier hatching, but these relationships were not consistent across all sites. Lower temperatures were also linked to increased spawn mortality. However, temperature increases were also strongly correlated with increases in urban area, arable area, and nitrate levels in the vicinity of spawning grounds. As with spawning and hatching, there was marked spatial variation in spawn mortality trends, where some sites exhibited steady increases over time in the proportion of dead or diseased spawn. These findings support previous work linking warming temperatures to shifts in timing of amphibian breeding, but also highlight the importance of assessing the effect of land use change and pollution on wild amphibian populations. These results have implications for our understanding of the response of wild amphibian populations to climate change, and the management of human-dominated landscapes for declining wildlife populations.

Development and optimization of a novel environmental DNA‐based method for moor frog (Rana arvalis) monitoring and comparison with a commercial detection kit

AbstractMoor frogs (Rana arvalis) are protected by the European Union's Habitats Directive, often making them a target for environmental impact assessments. However, moor frog detection with the traditional method using their mating calls is difficult, time‐consuming, and limited to the short mating season. Environmental DNA (eDNA)‐based methods could be beneficial in moor frog detection since moor frogs are shy and co‐occurring common frogs (R. temporaria) are morphologically similar. Additionally, eDNA‐based methods could even extend the detection period of moor frogs. We tested two different DNA isolation methods and two different eDNA detection methods, Sanger sequencing and quantitative PCR (qPCR), and compared the results to the traditional mating call survey method. In addition, we tested whether linear polyacrylamide treatment would improve eDNA detection. We sampled 27 sites in Finland, of which moor frogs were detected in 17 sites by traditional acoustic survey method, in 20 sites with Sanger sequencing‐based eDNA method, and in 21 sites with qPCR‐based eDNA method. We sampled water bodies during and 3–4 weeks after the end of the mating season. Our results show that both tested eDNA methods (Sanger sequencing and qPCR) can be used to detect moor frogs and extend the moor frog detection period by at least 3–4 weeks, but qPCR slightly outperforms Sanger sequencing. Linear polyacrylamide treatment did not improve the results. Detection probabilities were similar or higher with the eDNA‐based methods than with the traditional survey method, as moor frogs were also detected in water bodies in which mating calls were not heard. Based on our results, eDNA‐based detection methods can be highly beneficial in monitoring the presence of moor frogs and can be used to complement traditional surveys.

Amphibian survey of Ko Pha-gnan in Surat Thani Province, Thailand.

Insular amphibian species are often overlooked, rendering them vulnerable to habitat encroachment and other anthropogenic threats. The aim of this study was to compile a comprehensive list of amphibian species on Ko Pha-ngan in Surat Thani Province, Thailand. Data were collected via transect surveys and drift line fence traps in three different habitat types from February 2021 to September 2023. Our efforts detected 12 unique amphibian species in each of the three habitat types. The highest number of detections was observed in the Ko Pha-ngan-Than Sadet National Park protected areas. The common tree frog (Polypedatesleucomystax) and the common Asian toad (Duttaphrynusmelanostictus) were the two most abundantly found species on the island, whereas the Koh Tao caecilian (Ichthyophiskohtaoensis) and the newly described false Doria's fanged frog (Limnonectespseudodoriae) where the least commonly found species. In addition, Microhylaheymonsi and Fejervaryalimnocharis tadpoles were observed developing in high-salinity water bodies. Many species have shown a high tolerance in human-dominated landscapes. This study sheds light on the need for additional monitoring to better understand the dynamics of endemic species in addition to the impact tourism-driven development and habitat destruction has on a species with an insularly finite habitat.

Baseline haematological parameters in three common Australian frog species.

Amphibians are experiencing declines globally, with emerging infectious diseases as one of the main causes. Haematological parameters present a useful method for determining the health status of animals and the effects of particular diseases, but the interpretation of differential cell counts relies on knowing the normal ranges for the species and factors that can affect these counts. However, there is very little data on either normal haematological parameters or guides for blood cell types for free-ranging frog species across the world. This study aims to 1) create a visual guide for three different Australian frog species: Litoria paraewingi, Limnodynastes dumerilii, and Crinia signifera, 2) determine the proportions of erythrocytes to leukocytes and 3) differential leukocytes within blood smears from these three species and 4) assess the association between parasites and differential counts. We collected blood samples from free-ranging frogs and analysed blood smears. We also looked for ectoparasites and tested for the fungal disease chytridiomycosis. Overall, we found that the differentials of erythrocytes to leukocytes were not affected by species, but the proportions of different leukocytes did vary across species. For example, while lymphocytes were the most common type of leukocyte across the three species, eosinophils were relatively common in Limnodynastes dumerilii but rarely present in the other two species. We noted chytridiomycosis infection as well as ectoparasites present in some individuals but found no effect of parasites on blood parameters. Our results add baseline haematological parameters for three Australian frog species and provide an example of how different frog species can vary in their differential blood cell counts. More information is needed on frog haematological data before these parameters can be used to determine the health status of wild or captive frogs.

Морфологічна диференціація Anura (Amphibia) у пізньому онтогенезі: аналіз популяцій Rana temporaria з регіону Карпат

The study is devoted to the morphological features of the common frog of the Carpathian region of Ukraine, namely the expression, directionality and structure of sexual differences of this species. The work uses data obtained as a result of the processing of the scientific collection of Rana temporaria (Linnaeus, 1758) from the territory of the Ukrainian Carpathians (Lviv, Ivano-Frankivsk, and Zakarpattia oblasts), which is housed at the National Museum of Natural History, National Academy of Sciences of Ukraine (Kyiv). A total of 381 common frog specimens were studied—233 males (L = 24.6–84.5 mm) and 148 females (L = 23.2–94.2 mm). It is shown that individual differences in linear dimensions and body proportions in a sample of common frog males and females are structured and clearly reflect the morphological differentiation of individuals according to the age criterion. In the late ontogenesis of the common frog, generalised sex-related differences extend with the increasing age of individuals: in juveniles SqMD = 2.36, in subadults SqMD = 4.77, and in adults SqMD = 22.08. At the same time, not only does the expression of sex-related differences increase, but also their orientation changes significantly. According to our data, this year’s juveniles (juvenis) are individuals that did not winter at all (0+). At the same time, males are reliably (t = 4.50; p < 0.001) larger in body length (24.6–37.5 mm; Lmean = 29.87 mm) than females (23.2–32.7 mm; Lmean = 26.40 mm). But already in the group of semi-adult (subadultus) frogs, the orientation of sexual dimorphism in terms of body length changes completely—females are reliably larger than males, which is even more pronounced in adult, sexually mature individuals. Males of tailless amphibians grow and physiologically mature faster than females, so they begin to reproduce 1–2 years earlier. Females, which grow more slowly and mature later, join the reproduction having a larger size and older age. It is shown that sexual differences begin to form in the subadultus group. The generalised differences (SqMD) by sex are significantly smaller than by age. Adult sexually mature females compared to males are characterised by larger values of 18 body parameters, except for shoulder length (H.) and elbow joint diameter (Cr.a.c.), which are most reliably larger in males. These features of the proportions of the limbs provide the male with reliable holding of the female during mating (amplexus).

Географическая изменчивость длительности личиночного развития и размеров метаморфов травяных лягушек (Ranidae, Anura)

With the help of artificial inter-population crosses of adult Rana temporaria from four spatially remote populations of the European part of the species' range, the magnitude and direction of changes in metamorphic traits were estimated. The maximum values of the size of metamorphs at the end of metamorphosis and the rate of larval growth were found in the descendants of the parents of Belarusian populations (Turov and Minsk) and, as a rule, in hybrid offspring with the participation of the parents of these populations, when crossing with individuals from the population of the Moscow region (ZBS). Outbreeding depression of the time of larval development before the end of metamorphosis and growth rate was revealed when crossing ZBS females with Turov males. In a population from a region with a relatively low climatic temperature (Kirov), the minimum size of metamorphs was revealed, but not the time of larval development. In most crosses, a relatively greater contribution of non-additive genetic variability, as well as the maternal effect associated with the difference in egg sizes, to the formation of inter-population differences was revealed. The inter-population variability of studied traits related to fitness is often directed along the gradient of environmental conditions, and not counter-gradient (as should be expected based on the conception of counter-gradient selection), which is explained by the choice of different growth and development strategies in tadpoles formed in southern and northern populations.

Traffic and weather influence on small wildlife carcass persistence time on roads

The rapidly expanding road network threatens the persistence of many terrestrial species through mortality associated with wildlife-vehicle collisions. Roadkill monitoring under-estimate actual collision numbers as the time during which roadkill carcasses remain visible on the road is often shorter than the frequency of road monitoring. By placing passerine and amphibian carcasses on stretches of roads we surveyed every 2 h, we fine-tuned existing persistence estimates for these species. The time for half of the carcasses to disappear was less than 30 min for birds and 1–18 h for amphibians depending on the volume of traffic, which is much shorter than previous estimates. We show the implications of our results by performing roadkill surveys on common toads 3 h after the window of reproductive migration, showing roadkill is under-estimated by half, and by estimating from citizen-science data that passerine collisions could be as high as 10 individuals.km-2.year-1 in south-eastern France.

Movement behaviour of the common frog (Rana temporaria) in an anthropized mountain landscape: integrated step selection analysis of GPS tracking data

Abstract Movement is a key aspect of amphibian ecology as the life cycle of many species involves seasonal dispersal and migration. However, there is still a lack of knowledge about how amphibians move across landscapes in relation to landscape structure and resource availability. Modern lightweight GPS loggers and associated statistical tools offer new perspectives to fill this gap. We tracked the movements of 20 adult European common frogs (Rana temporaria) in a human-transformed mountain landscape during summer using GPS tags programmed with a constant 4-hour time interval between fixes. Using integrated step-selection analysis, we demonstrate firstly that frogs strongly selected aquatic habitats over open grassland. Second, ski runs have a negative effect on frogs, indicated by longer movement steps. Weather (air temperature and rain) had no effect on frog movements. These results constitute a promising first step towards studying anuran movement in relation to landscape structure at fine spatial and temporal scales.

Body size mediates latitudinal population differences in the response to chytrid fungus infection in two amphibians

Factors behind intraspecific variation in sensitivity to pathogens remain poorly understood. We investigated how geographical origin in two North European amphibians affects tolerance to infection by the chytrid fungus Batrachochytrium dendrobatidis (Bd), a generalist pathogen which has caused amphibian population declines worldwide. We exposed newly metamorphosed individuals of moor frog Rana arvalis and common toad Bufo bufo from two latitudinal regions to two different BdGPL strains. We measured survival and growth as infections may cause sub-lethal effects in fitness components even in the absence of mortality. Infection loads were higher in B. bufo than in R. arvalis, and smaller individuals had generally higher infection loads. B. bufo had high mortality in response to Bd infection, whereas there was little mortality in R. arvalis. Bd-mediated mortality was size-dependent and high-latitude individuals were smaller leading to high mortality in the northern B. bufo. Bd exposure led to sub-lethal effects in terms of reduced growth suggesting that individuals surviving the infection may have reduced fitness mediated by smaller body size. In both host species, the Swedish Bd strain caused stronger sublethal effects than the British strain. We suggest that high-latitude populations can be more vulnerable to chytrids than those from lower latitudes and discuss the possible mechanisms how body size and host geographical origin contribute to the present results.

Special Nuclear Structures in the Germinal Vesicle of the Common Frog with Emphasis on the So-Called Karyosphere Capsule.

The karyosphere (karyosome) is a structure that forms in the oocyte nucleus-germinal vesicle (GV)-at the diplotene stage of meiotic prophase due to the assembly of all chromosomes in a limited portion of the GV. In some organisms, the karyosphere has an extrachromosomal external capsule, the marker protein of which is nuclear F-actin. Despite many years of theories about the formation of the karyosphere capsule (KC) in the GV of the common frog Rana temporaria, we present data that cast doubt on its existence, at least in this species. Specific extrachromosomal strands, which had been considered the main elements of the frog's KC, do not form a continuous layer around the karyosphere and, according to immunogold labeling, do not contain structural proteins, such as actin and lamin B. At the same time, F-actin is indeed noticeably concentrated around the karyosphere, creating the illusion of a capsule at the light microscopy/fluorescence level. The barrier-to-autointegration factor (BAF) and one of its functional partners-LEMD2, an inner nuclear membrane protein-are not localized in the strands, suggesting that the strands are not functional counterparts of the nuclear envelope. The presence of characteristic strands in the GV of R. temporaria late oocytes may reflect an excess of SMC1 involved in the structural maintenance of diplotene oocyte chromosomes at the karyosphere stage, since SMC1 has been shown to be the most abundant protein in the strands. Other characteristic microstructures-the so-called annuli, very similar in ultrastructure to the nuclear pore complexes-do not contain nucleoporins Nup35 and Nup93, and, therefore, they cannot be considered autonomous pore complexes, as previously thought. Taken together, our data indicate that traditional ideas about the existence of the R. temporaria KC as a special structural compartment of the GV are to be revisited.

Toxicity assessment of a tebuconazole-based fungicide on the embryo-larval development of the common south American toad Rhinella arenarum.

Agrochemicals cause diverse effects on aquatic communities, and amphibian species are particularly threatened due the high susceptibility to contamination. Present study evaluates the toxicity of a widely used fungicide tebuconazole (Trigal®) by the assessment of mortality and developmental alterations at acute, subchronic, and chronic exposure during the embryo-larval development of the South American toad Rhinella arenarum. Also, the sensitivity of the different embryonic stages was evaluated with 24-h pulse exposure treatments. The results demonstrated that larvae were more sensitive than embryos at acute exposure (LC50-24 and 96 h = 74.62, 31.92 mg/L and 24.27, 16.81 mg/L for embryos and larvae, respectively). Nevertheless, embryos toxicity increased significantly achieving a sensitivity very similar to larvae at chronic exposure (LC50-168 and 504 h = 13.31, 4.35 mg/L and 14.47, 6.83 mg/L for embryos and larvae, respectively). Embryos exhibited several sublethal effects from 5 mg/L at 96 h onwards, such as delayed development, reduce body size, edemas, tail/axial flexures, weakness, and absence of movements. The teratogenic index at 96 h was 10.13, indicating the severe teratogenic potential of the fungicide. 24-h pulse exposure treatments showed an increased sensitivity in intermediate stages as S.11, S.18, S20, and S.23 (NOEC-96 h = 100, 200, 75, and 20 mg/L, respectively), while stage S.25 was the most sensitive to the fungicide (NOEC-96 h = 5 mg/L). About metamorphic process, tebuconazole caused an acceleration of metamorphosis at the lowest concentration (0.001 mg/L), but also an increase in mortality and in addition, significant differences in the weight in all treatments. The results obtained throughout this work indicate that tebuconazole cause several adverse effects in Rhinella arenarum embryo-larval development.

A novel tamanavirus (Flaviviridae) of the European common frog (Rana temporaria) from the UK.

Flavivirids are small, enveloped, positive-sense RNA viruses from the family Flaviviridae with genomes of ~9-13 kb. Metatranscriptomic analyses of metazoan organisms have revealed a diversity of flavivirus-like or flavivirid viral sequences in fish and marine invertebrate groups. However, no flavivirus-like virus has been identified in amphibians. To remedy this, we investigated the virome of the European common frog (Rana temporaria) in the UK, utilizing high-throughput sequencing at six catch locations. De novo assembly revealed a coding-complete virus contig of a novel flavivirid ~11.2 kb in length. The virus encodes a single ORF of 3456 aa and 5' and 3' untranslated regions (UTRs) of 227 and 666 nt, respectively. We named this virus Rana tamanavirus (RaTV), as BLASTp analysis of the polyprotein showed the closest relationships to Tamana bat virus (TABV) and Cyclopterus lumpus virus from Pteronotus parnellii and Cyclopterus lumpus, respectively. Phylogenetic analysis of the RaTV polyprotein compared to Flavivirus and Flavivirus-like members indicated that RaTV was sufficiently divergent and basal to the vertebrate Tamanavirus clade. In addition to the Mitcham strain, partial but divergent RaTV, sharing 95.64-97.39 % pairwise nucleotide identity, were also obtained from the Poole and Deal samples, indicating that RaTV is widespread in UK frog samples. Bioinformatic analyses of predicted secondary structures in the 3'UTR of RaTV showed the presence of an exoribonuclease-resistant RNA (xrRNA) structure standard in flaviviruses and TABV. To examine this biochemically, we conducted an in vitro Xrn1 digestion assay showing that RaTV probably forms a functional Xrn1-resistant xrRNA.

Аналіз спектру живлення жаби трав’яної (Rana temporaria) та ропухи сірої (Bufo bufo) в антропогенно-трансформованих оселищах Українського Розточчя

As a result of the research, 109 individuals of two amphibian species: the common frog (Rana temporaria) and the common toad (Bufo bufo) were selected. These two species are the most numerous among other species of amphibians within the studied territory, therefore, they can be considered typical species in the anthropogenically transformed habitats of the Ukrainian Roztochchia. During the analysis of the collected material, Ivlev’s selectivity index was used to determine the selective attitude of the studied amphibian species to the number of consumed objects. Comparing data obtained from Barber’s traps and material extracted from the stomachs of amphibians, it was concluded that depending on the habitat type, both the common frog (Rana temporaria) and the common toad (Bufo bufo) show a certain feeding selectivity. The common toad consumes mostly the representatives of Annelida (15%), Arachnida (15%) and Coleoptera (18%), what makes up 48% of all consumed objects in total. The common frog, under the conditions of anthropogenically transformed environment, prefers Mollusca (15%), Myriapoda (20%) and Hemiptera (20%) mostly, what is 55% in total. In half of the stomachs we examined for, both amphibian species, plant residues were found, specifically parts of leaves and branches. Additionally, in individuals of the Bufo bufo species, stones were discovered in the stomach. Most likely, these items ended up there accidentally during the capture of primary food objects. Species of tailless amphibians are highly vulnerable to the negative impact of various anthropogenic factors. Deforestation, water bodies drainage, and global or local environmental pollution can significantly affect diverse ecosystems. This, in turn, may lead to the disappearance of segments of the dietary spectrum for amphibians. Monitoring the trophic interactions of this animal class is crucial, given the importance of amphibians as vital links in food chains. This factor underscores the need for periodic trophic monitoring, as amphibians play a pivotal role in sustaining ecological balance within their vulnerable populations.

Colouration matters in dull toads: ultraviolet adornment for ladies and agrochemicals fading effects.

Integument colouration can influence many aspects of fitness, and is under strong sexual selection. Amphibians often express sexual dichromatism, and ultra-violet (UV) colouration is usually biased toward males as a sexual signal. As an honest signal, colouration is related to several individual traits, but can also be related to environmental factors such as anthropogenic pollutants, to which amphibians are highly sensitive. In this study, we investigated sexual dichromatism and UV reflectance covering a large visual spectrum (wavelength ranging from 300 to 700nm) on different body areas (throat, ventral and dorsal areas), in a widespread amphibian species, the spiny toad (Bufo spinosus). Then, we tested the impact of chronic exposure to two widespread herbicides (glyphosate's primary metabolite [AMPA] and Nicosulfuron) on their colouration. We found a strong but unexpected sexual dichromatism with females reflecting more in the UV spectrum (throat and ventral area) than males, suggesting these body parts might be critical in intra-specific signalling. Females with higher ventral UV reflectance were in better body condition, suggesting an honest signal role of UV reflectance which could influence male choice. Throat colouration was further differentially influenced by agrochemicals according to sexes. In AMPA-exposed males, throat was more saturated in yellow-orange than in control males, and Nicosulfuron exposure decreased the throat's reflectance hue in females, which can bear consequences on mate attractiveness. Future studies need to investigate the underlying mechanisms that are altered by agrochemical exposure.

Geographical Variation in Body Size in the Asian Common Toad (Duttaphrynus melanostictus).

The geographic variation in life-history traits of organisms and the mechanisms underlying adaptation are interesting ideas in evolutionary biology. This study investigated age and body size of the Asian common toad (Duttaphrynus melanostictus) among five populations along a geographical gradient. We found that geographical variation in age was non-significant among populations but there was a significant and positive correlation between mean age and body size. Although the body size values at 1043 m are quite different from other sites, after controlling for age effects, there was a significant positive correlation between altitude and body size. Our findings followed the predictions of Bergmann's rule, suggesting that the body size of D. melanostictus is potentially influenced by the low air temperatures at higher altitudes.

Evidence for Expensive Tissue Hypothesis in the Asian Common Toad (<italic>Duttaphrynus melanostictus</italic>)

Evidence for Expensive Tissue Hypothesis in the Asian Common Toad (<italic>Duttaphrynus melanostictus</italic>)

Perception of con- and heterospecific injury cues in tadpoles of dendrobatid, ranid and bufonid frogs (Anura)

Predator-prey interactions are vital for organismal survival. They shape anti-predator mechanisms and often depend on sensory abilities. Tadpoles use chemical cues, such as injury cues (alarm cues), to assess predation risks and modify their life-history, morphology, and behaviours accordingly. However, the prevalence of chemically mediated anti-predator responses in species with distinct ecological niches (e.g. within phytotelmata) remains unknown, hindering our understanding of the ecological significance and evolution of alarm substances. Therefore, our study aimed to investigate chemically mediated anti-predator responses in tadpoles of two Neotropical poison dart frogs, Ranitomeya sirensis and Epipedobates anthonyi (and compare their responses to two Palearctic model organisms, Rana temporaria and Bufo bufo, which are known to utilise alarm substances). Through behavioural bioassays, we exposed predator-naïve tadpoles to extracts of each species (i.e. con- and heterospecific cues), including water as a control (i.e. five treatments per species). We assessed changes in their activity before and after stimulus introduction. Our results show that E. anthonyi did not respond to any of the stimuli, whereas R. sirensis displayed increased activity levels exclusively in response to conspecific cues, but not to heterospecific cues. With this, our findings suggest a specialized recognition system in R. sirensis, potentially directed at conspecific competitors but likely unrelated to anti-predator mechanisms. In contrast, E. anthonyi may be insensitive to injury cues or utilize alternative sensory modalities to respond to acute predation events. This study sheds light on the chemical alarm response system of Neotropical poison dart frog tadpoles, providing foundational understanding of how dendrobatids react to injury cues. It prompts questions about the ecological significance and evolutionary implications of chemical communication in species facing extreme resource limitation during development and underscores the importance of comparative research for understanding chemical communication in diverse aquatic ecosystems.

OCCURRENCE OF PATHOGENIC CHYTRID FUNGI BATRACHOCHYTRIUM SALAMANDRIVORANS AND BATRACHOCHYTRIUM DENDROBATIDIS IN THE HONG KONG NEWT (PARAMESOTRITON HONGKONGENSIS) AND OTHER WILD AND IMPORTED AMPHIBIANS IN A SUBTROPICAL ASIAN REGION.

One of the major threats for the massive loss in global amphibian diversity is chytridiomycosis, caused by chytrid fungi Batrachochytrium dendrobatidis (Bd) and B. salamandrivorans (Bsal). Following its discovery in 2013, Bsal has emerged as a severe threat to the global survival of urodelans. In 2018, a study reported a high prevalence of Bsal (65.6%) in the Hong Kong newts (Paramesotriton hongkongensis, Near Threatened) of a southern China population adjacent to Hong Kong (HK). Uncertainty regarding the Bsal infection status of P. hongkongensis inhabiting HK raised deep concern over the risk of introducing Bsal from that population. We screened the skin swabs from wild individuals of P. hongkongensis, 15 sympatric amphibian species, and 16 imported amphibian species in HK for chytrids. We found that both Bsal and Bd occur in low prevalences in P. hongkongensis (Bsal 1.7%, 5/293; Bd 0.34%, 1/293), Hong Kong cascade frog, Amolops hongkongensis, family Ranidae (Bsal only, 5.26%, 1/19), and Asian common toad, Duttaphrynus melanostictus, family Bufonidae (Bsal only, 5.88%, 1/17), populations of HK, with infected individuals being asymptomatic, suggesting a potential role of these species as reservoirs of Bsal. Conversely, Bd, but not Bsal, was present on 13.2% (9/68) of imported amphibians, indicating a high chytrid introduction risk posed by international amphibian trade. Long-term surveillance of the presence of Bd and Bsal in wild and captive amphibians would be advisable, and we recommend that import and export of nonnative chytrid carriers should be prevented, especially to those regions with amphibian populations naïve to Bd and Bsal.