Search Results (14)

Mitochondrial genomes (mitogenomes) play a key role in species identification and phylogenetic studies due to their stable gene arrangements and evolutionary insights. Dryophytes flaviventris, classified in 2020 and closely related to D. suweonensis, lacks mitochondrial DNA data for differentiation. This gap hinders accurate species identification, highlighting the need for further genomic studies. The complete mitogenome size of two D. flaviventris were 18,616–18,617 bp and those for two D. suweonensis were 18,610–18,616 bp, the mitogenomes of the two species consisting of 13 protein-coding genes (PCGs), two ribosomal RNA genes, 22 transfer RNA (tRNA) genes, and a D-loop. Phylogenetic analysis confirmed that the mitochondrial DNA of all four individuals formed a monophyletic group, showing no genetic differentiation. As a result, the two species do not form distinct clades, and mitogenomes could not differentiate them, suggesting they are not reciprocally monophyletic. This study presents the first mitogenome data for D. flaviventris and provides valuable insights into tree frog taxonomy. Full article

The amphibian chytrid fungus, Batrachochytrium dendrobatidis (Bd), has been implicated as an agent of acute declines in amphibian populations worldwide. East Asian amphibians have been coexisting with Bd for long periods and thus are considered resistant; among the many is the Japanese tree frog, Dryophytes japonicus. Our study focused Bd infection effects on reproductive behaviors and physiological parameters in D. japonicus as a function of better understanding the chronic effect of the disease on long-term population viability. During the peak breeding season, we captured 70 males and quantified the chorus size, calling behaviors, physiological states, innate immunity, and sperm quality of individuals. In a simple comparison, all parameters were not significantly different. However, in the NMDS analysis, we were able to confirm subtle trends in some items according to infection and correlations between several items. Importantly, sperm density and sperm vitality tend to increase with Bd infection load, suggesting increased reproductive effort following infection. Additionally, this analysis indicated that innate immunity was positively related to Bd infection intensities, indicating the activation of immunity upon infection. These findings indicate that Bd-resistant D. japonicus maintains reproductive capabilities and physio-logical stability despite Bd infection, likely due to a co-evolved immune system. The present work offers insight into how amphibian populations may have some endurance in the presence of Bd and points out the importance of studying resistant species as a means to understand long-term ecological effects. Our results suggest that resistance to Bd may not simply prevent cata-strophic declines but actively contribute to the dynamics of Bd prevalence in amphibian communities, and confer implications for conservation strategies. Full article

The grey tree frog, Dryophytes versicolor, survives whole-body freezing for weeks during cold winter months. Survival in a state devoid of available food, water, or oxygen forces a reliance on metabolic rate depression (MRD) and the reprioritization of bodily functions. This study utilizes next-generation sequencing (NGS) and bioinformatic analyses to characterize changes in the microRNAome of D. versicolor. When comparing control to frozen groups, five microRNAs (miRNA) were found to be differentially regulated (miR-143-3p, miR-30e-3p, miR-10a-5p, miR-140-3p, and miR-148a-3p), suggesting that they play key roles in freeze survival. The KEGG and GO analyses of these changes predicted a significant negative enrichment of terms associated with cell proliferation and active metabolism while simultaneously predicting the upregulation of cell signalling terms. These results suggest a fast-acting regulatory role for miRNA in contributing to the reorganization of gene expression and the limitation of energy-expensive processes during MRD in the hind leg skeletal muscle of the frog. Full article

Extreme weather poses huge challenges for animals that must adapt to wide variations in environmental temperature and, in many cases, it can lead to the local extirpation of populations or even the extinction of an entire species. Previous studies have found that one element of amphibian adaptation to environmental stress involves changes in mitochondrial gene expression at low temperatures. However, to date, comparative studies of gene expression in organisms living at extreme temperatures have focused mainly on nuclear genes. This study sequenced the complete mitochondrial genomes of five Asian hylid frog species: Dryophytes japonicus, D. immaculata, Hyla annectans, H. chinensis and H. zhaopingensis. It compared the phylogenetic relationships within the Hylidae family and explored the association between mitochondrial gene expression and evolutionary adaptations to cold stress. The present results showed that in D. immaculata, transcript levels of 12 out of 13 mitochondria genes were significantly reduced under cold exposure (p < 0.05); hence, we put forward the conjecture that D. immaculata adapts by entering a hibernation state at low temperature. In H. annectans, the transcripts of 10 genes (ND1, ND2, ND3, ND4, ND4L, ND5, ND6, COX1, COX2 and ATP8) were significantly reduced in response to cold exposure, and five mitochondrial genes in H. chinensis (ND1, ND2, ND3, ND4L and ATP6) also showed significantly reduced expression and transcript levels under cold conditions. By contrast, transcript levels of ND2 and ATP6 in H. zhaopingensis were significantly increased at low temperatures, possibly related to the narrow distribution of this species primarily at low latitudes. Indeed, H. zhaopingensis has little ability to adapt to low temperature (4 °C), or maybe to enter into hibernation, and it shows metabolic disorder in the cold. The present study demonstrates that the regulatory trend of mitochondrial gene expression in amphibians is correlated with their ability to adapt to variable climates in extreme environments. These results can predict which species are more likely to undergo extirpation or extinction with climate change and, thereby, provide new ideas for the study of species extinction in highly variable winter climates. Full article

The calling behavior of anurans should be studied in detail as it greatly influences their physiology and immunity, particularly in prolonged breeding species. The effect can be further complicated by the emergence timing in the breeding season. We conducted a study comparing the physiology and calling behavior of the Japanese tree frog (Dryophytes japonicus), a prolonged breeder species, according to the breeding timing. During the middle of the breeding season, a high chorus size appeared, indicating a breeding peak. However, chorus size did not dominate physiology and calling behavior. In the early breeding season, frogs had a high energy storage state and immunity. In the middle of the breeding season, individuals from the early breeding season were considered to have exhausted their energy stores and had low immunity. Towards the end of the breeding season, frogs appeared to have newly introduced, at which time energy stores and immunity were as high as in the beginning. However, unlike the physiology, the pattern of calling constantly varied as the breeding season progressed. Frogs from the early season conserved energy used for calling, and frogs from the late season showed a breeding spurt for mating. Our results can help in understanding the energy metabolism of calling behavior, physiology, and disease epidemiology in prolonged breeder species. They also suggest that individuals coordinate their participation in the breeding season and that the timing of their appearance at breeding sites may not be random. Full article

Hyla sanchiangensis (Anura: Hylidae) is endemic to China and is distributed across Anhui, Zhejiang, Fujian, Guangdong, Guangxi, Hunan, and Guizhou provinces. The mitogenomes of H. sanchiangensis from two different sites (Jinxiu, Guangxi, and Wencheng, Zhejiang) were sequenced. Phylogenetic analyses were conducted, including 38 mitogenomes of Hylidae from the NCBI database, and assessed the phylogenetic relationship of H. sanchiangensis within the analyzed dataset. Two mitogenomes of H. sanchiangensis showed the typical mitochondrial gene arrangement with 13 protein-coding genes (PCGs), two ribosomal RNA genes (12S rRNA and 16S rRNA), 22 transfer RNA (tRNA) genes, and one non-coding control region (D-loop). The lengths of the 12S rRNA and 16S rRNA genes from both samples (Jinxiu and Wencheng) were 933 bp and 1604 bp, respectively. The genetic distance (p-distance transformed into percent) on the basis of the mitogenomes (excluding the control region) of the two samples was calculated as 4.4%. Hyla sanchiangensis showed a close phylogenetic relationship with the clade of (H. annectans + H. tsinlingensis), which was supported by ML and BI analyses. In the branch-site model, five positive selection sites were found in the clade of Hyla and Dryophytes: Cytb protein (at position 316), ND3 protein (at position 85), and ND5 protein (at position 400) have one site, respectively, and two sites in ND4 protein (at positions 47 and 200). Based on the results, we hypothesized that the positive selection of Hyla and Dryophytes was due to their experience of cold stress in historical events, but more evidence is needed to support this conclusion. Full article

To cope with amphibian die-offs caused by ranavirus, it is important to know the underlying ranavirus prevalence in a region. We studied the ranavirus prevalence in tadpoles of two native and one introduced anuran species inhabiting agricultural and surrounding areas at 49 locations across eight provinces of South Korea by applying qPCR. The local ranavirus prevalence and the individual infection rates at infected locations were 32.6% and 16.1%, respectively, for Dryophytes japonicus (Japanese tree frog); 25.6% and 26.1% for Pelophylax nigromaculatus (Black-spotted pond frog); and 30.5% and 50.0% for Lithobates catesbeianus (American bullfrog). The individual infection rate of L. catesbeianus was significantly greater than that of D. japonicus. The individual infection rate of P. nigromaculatus was related to the site-specific precipitation and air temperature. The individual infection rate gradually increased from Gosner development stage 39, and intermittent infection was confirmed in the early and middle developmental stages. Our results show that ranavirus is widespread among wild amphibians living in agricultural areas of South Korea, and mass die-offs by ranavirus could occur at any time. Full article

The number of science projects with citizen (volunteer) participants is increasing nowadays. Despite some advantages citizen science have which directly influence science fields, the largest weakness of citizen science is the issue of the reliability of data collected by non-specialists. In this study, we assess the reliability of data collected by citizens and identify the factors that impede the credibility. We selected two species, Pelophylax chosenicus and Dryophytes suweonensis, which have specific habitat, breeding and brumation positions. The location data for both species were collected by the global citizen engagement project “iNaturalist”. As a result, 89.3% for P. chosenicus and 37.1% for D. suweonensis of the location data were estimated to be erroneous. No difference was observed in the data accuracy between experts and citizens for D. suweonensis because the mating call is the main classification key for the species. On the other hand, a significant difference was confirmed in the case of P. chosenicus, whose external characteristics are its classification key. Our study shows that efforts to improve reliability, including appropriate species selection and survey methods, are necessary to use citizen participation data in science research. Full article

As the advantages of wind energy as an eco-friendly strategy for power generation continue to be revealed, the number of offshore wind farms also increases worldwide. However, wind turbines can induce behavioral and physiological responses in animals by emitting various types of noises. In this study, we investigated the behavioral, physiological, and immunological responses of male Japanese tree frogs (Dryophytes japonicus) when exposed to wind turbine noise. To determine the effects during the breeding season, frogs were collected from areas with and without wind turbines. Additionally, we exposed the frogs to recorded wind turbine noise at a site without a wind generator for 1 h to 24 h to analyze the short-term effects. Three types of calling patterns (dominant frequency, note duration, and call rate) were analyzed to investigate behavioral responses. Physiological responses were assessed using two steroid hormones assays, namely testosterone and corticosterone detection in the saliva. The immunity of each individual was assessed using a bacterial killing assay. The wind turbine group in the field had a higher call rate and corticosterone levels and lower immunity than the group in the field without turbines present, and all three of these variables were correlated with each other. Conversely, in the noise exposure experiment, a higher call rate was only observed post-exposure compared to pre-exposure. Thus, turbine noise seems to induce decreased immunity in Japanese tree frogs as an increase in energy investment that triggers a behavioral response rather than acting as a sole physiological response that leads to a direct increase in corticosterone. This decreased immunity due to energy tradeoff or physiological response can change the disease epidemiology of the population and create new adaptive patterns in these habitats. Full article

Determining the range, status, ecology and behaviour of species from areas where surveys and samplings are uncommon or difficult to conduct is a challenge, such as in the Democratic People’s Republic of Korea (DPR Korea). Here, we used genetic samples, field surveys, call recordings, photographic identification and a literature review to estimate the presence, range and status of amphibians in the DPR Korea. From our combined results and based on the IUCN Red List categories and criteria, we were able to estimate the national threat levels for most species. Our results demonstrated the presence of 18 native species and the suspected presence of Karsenia koreana and two Onychodactylus species. We reported the first record for Rana uenoi in the vicinity of Pyongyang using molecular tools and similarly confirmed the presence of Dryophytes japonicus at the same location. Based on distribution and modelling, we can expect the contact zone between species within the Rana and Onychodactylus genera to be located along the Changbai Massif, a mountain range that marks a shift in ecoregions and acts as a barrier to dispersion. The species richness was higher in the lowlands and at lower latitudes, with such areas populated by up to 11 species, while more northern regions were characterised by species richness of about half of that value. The combination of ecological models and known threats resulted in the recommendation of ten species as threatened at the national level following the IUCN Red List categories and criteria. This high number of threatened species was anticipated based on the high threat level to amphibians in bordering nations and globally. While the ecology of species in the DPR Korea is still understudied, we argue that species relying on agricultural wetlands such as rice paddies are not under imminent threat due to the enduring presence of extensive agricultural landscapes with low rates of chemical use and mechanisation. The maintenance of such landscapes is a clear benefit to amphibian species, in contrast to more industrialised agricultural landscapes in neighbouring nations. In comparison, the status of species dependent on forested habitats is unclear and threat levels are likely to be higher because of deforestation, as in neighbouring nations. Full article

Numerous amphibian species are declining because of habitat loss and fragmentation due to urbanization of landscapes and the construction of roads. This is a mounting threat to species restricted to habitats close to urban areas, such as agricultural wetlands in North East Asia. The Suweon treefrog (Dryophytes suweonensis) falls into the list of species threatened with habitat loss and most populations are under threat of extirpation. Over the last decades, sub-populations have become increasingly disconnected and specifically the density of paved roads has increased around the only site connecting northern and southern Seoul populations. We surveyed this locality in Hojobeol, Siheung, Republic of Korea in 2012, 2015 and 2019 to first confirm the decline in the number of sites where D. suweonensis was present. The second objective was to analyze the habitat characteristics and determine the remaining suitable habitat for D. suweonensis through a species distribution model following the maximum entropy method. Our results show that rice paddy cover and distance from the paved road are the most important factor defining suitable habitat for D. suweonensis. At this locality, uninterrupted rice paddies are a suitable habitat for the species when reaching at least 0.19 km², with an average distance of 138 ± 93 m² from the roads. We link the decrease in the number of sites where D. suweonensis is present with the decrease in rice paddy cover, generally replaced by localized infrastructures, greenhouses and habitat fragmentation. Rice paddies should remain connected over a large area for the protection of the remaining populations. In addition, habitat requirements should be integrated in the requisites to designate protected areas. Full article

Threat assessment is important to prioritize species conservation projects and planning. The taxonomic resolution regarding the status of the “Dryophytes immaculatus group” and the description of a new species in the Republic of Korea resulted in a shift in ranges and population sizes. Thus, reviewing the IUCN Red List status of the three species from the group: D. immaculatus, D. suweonensis and D. flaviventris and recommending an update is needed. While the three species have similar ecological requirements and are distributed around the Yellow Sea, they are under contrasting anthropological pressure and threats. Here, based on the literature available, I have applied all IUCN Red List criterion and tested the fit of each species in each criteria to recommend listing under the appropriate threat level. This resulted in the recommendation of the following categories: Near Threatened for D. immaculatus, Endangered following the criteria C2a(i)b for D. suweonensis and Critically Endangered following the criteria E for D. flaviventris. All three species are declining, mostly because of landscape changes as a result of human activities, but the differences in range, population dynamics and already extirpated subpopulations result in different threat levels for each species. Dryophytes flaviventris is under the highest threat category mostly because of its limited range segregated into two subpopulations; and several known extirpated subpopulations. Immediate actions for the conservation of this species are required. Dryophytes suweonensis is present in both the Republic of Korea and the Democratic Republic of Korea (DPR Korea) and is under lower ecological pressure in DPR Korea. Dryophytes immaculatus is present in the People’s Republic of China, over a very large range despite a marked decline. I recommend joint efforts for the conservation of these species. Full article

Amphibians are in the midst of a sixth mass extinction, and human activities play a major role in pushing species towards extinction. Landscape anthropisation has impacts that indirectly threaten species, in addition to the obvious destruction of natural habitats. For instance, land modification may bring human-commensal species in contact with sister-clades from which they were previously isolated. The species in these new contact zones are then able to hybridise to the point of reaching lineage fusion, through which the gene pool of the two species merges and one of the parental lineages becomes extirpated. Here, we documented the patterns of hybridisation between the spatially restricted D. suweonensis and the widespread D. japonicus. On the basis of the analysis of Cytochrome c oxidase subunit I mitochondrial DNA sequences (404 individuals from 35 sites) and six polymorphic microsatellites (381 individuals from 34 sites), we revealed a generalised, bi-directional, and geographically widespread hybridisation between the two species. Evidence of fertile back-crosses is provided by relatively high numbers of individuals in cyto-nuclear disequilibrium, as well as the presence of hybrid individuals further south than the species distribution limit, determined on the basis of call properties. Hybridisation is an additional threat to the endangered D. suweonensis. Full article

Amphibian skin secretions are enriched with complex cocktails of bioactive molecules such as proteins, peptides, biogenic amines, alkaloids guanidine derivatives, steroids and other minor components spanning a wide spectrum of pharmacological actions exploited for centuries in folk medicine. This study presents evidence on the protein profile of the skin secretions of the canyon tree frog, Dryophytes arenicolor. At the same time, it presents the reverse-phase liquid chromatography isolation, mass spectrometry characterization and identification at mRNA level of a novel 58 amino acids Kunitz-like polypeptide from the skin secretions of Dryophytes arenicolor, arenin. Cell viability assays performed on HDFa, CaCo2 and MCF7 cells cultured with different concentrations of arenin showed a discrete effect at low concentrations (2, 4, 8 and 16 µg/mL) suggesting a multi-target interaction in a hormetic-like dose-response. Further work is required to investigate the mechanisms underlying the variable effect on cell viability produced by different concentrations of arenin. Full article