Amphibian Species Research Articles

New records of amphibians from Ha Tinh Province, Vietnam.

Since the establishment of the Vu Quang National Park in 2002 in Ha Tinh Province, central Vietnam, several surveys on the amphibian fauna have been undertaken in this protected area. In 2009, Nguyen et al. provided a list of 23 amphibian species from Vu Quang National Park. In addition, a new species was described in 2021 from the National Park, namely Vietnamophrynevuquangensis. As a result of our field surveys in 2013 and 2023, a total of 29 species of amphibians was recorded from the Vu Quang National Park. Six of them are recorded for the first time from Ha Tinh Province, comprising three species of Megophryidae, one species of Dicroglossidae, one species of Ranidae and one species of Rhacophoridae. In addition, we provide morphological data and ecological notes on the aforementioned species.

Species composition and conservation of amphibians in Central Panay Mountain Range, Antique, Philippines

Abstract. Flores GMA, Torres AG, Flores ABA, Jakosalem PGC, Guihawan JQ, Roa-Quiaoit HA. 2024. Species composition and conservation of amphibians in Central Panay Mountain Range, Antique, Philippines. Biodiversitas 25: 1797-1808. Assessing amphibian species composition and conservation status in Central Panay Mountain Range (CPMR), the largest contiguous forest in West Visayas in central Philippines, provides relevant and updated information on the status and conservation needs of the amphibians. Opportunistic, visual encounter and acoustic surveys were employed across diverse habitats-lowland and lower montane forests, streams, and grasslands. A total of 11 amphibian species from six families were documented, with notable findings including endemic species such as Rough-backed forest frog Platymantis corrugatus Duméril 1853, Dumeril’s wrinkled ground frog Platymantis dorsalis Duméril 1853, and Philippine narrow-mouth toad Kaloula conjuncta negrosensis Taylor 1922, as well as threatened species Panay forest frog Platymantis panayensis Brown, Brown and Alcala 1997, which is endemic to Panay Island. The P. dorsalis emerged as the most abundant, with 127 individuals recorded. Furthermore, two new distribution records of amphibians, narrow-mouth toad Kaloula sp. and panther flying frog Rhacophorus pardalis Günther 1858 were identified, potentially representing previously undocumented species endemic to the Philippines. The survey also revealed local threats posing significant risks to amphibian populations in the area. Several conservation measures have been proposed to protect amphibians and their habitats within CPMR. Given the area's rich biodiversity and ecological integrity, efforts are currently being implemented to establish CPMR as a nationally protected area, and this study contributes to this initiative by providing baseline data on amphibian diversity, underscoring the role of amphibians as ecological indicators of ecosystem health.

Building pondscapes for amphibian metapopulations.

The success of ponds constructed to restore ecological infrastructure for pond-breeding amphibians and benefit aquatic biodiversity depends on where and how they are built. We studied effects of pond and landscape characteristics, including connectivity, on metapopulation dynamics of 12 amphibian species in Switzerland. To understand the determinants of long-term occupancy (here summarized as incidence), environmental effects on both colonization and persistence should be considered. We fitted dynamic occupancy models to 20years of monitoring data on a pond construction program to quantify effects of pond and landscape characteristics and different connectivity metrics on colonization and persistence probabilities in constructed ponds. Connectivity to existing populations explained dynamics better than structural connectivity metrics, and simple metrics (distance to the nearest neighbor population, population density) were useful surrogates for dispersal kernel-weighted metrics commonly used in metapopulation theory. Population connectivity mediated the persistence of conservation target species in new ponds, suggesting source-sink dynamics in newly established populations. Population density captured this effect well and could be used by practitioners for site selection. Ponds created where there were 2-4 occupied ponds within a radius of ∼0.5km had >3.5 times higher incidence of target species (median) than isolated ponds. Species had individual preferences regarding pond characteristics, but breeding sites with larger (≥100m2) total water surface area, that temporarily dried, and that were in surroundings with maximally 50% forest benefitted multiple target species. Pond diversity will foster amphibian diversity at the landscape scale.

Groundwater decline has negatively affected the well-preserved amphibian community of Doñana National Park (SW Spain)

Abstract The success of amphibian conservation efforts is highly dependent on the preservation of amphibian breeding habitats. Doñana National Park contains an abundance of ponds with heterogeneous characteristics, which has historically favoured its amphibian community. However, most of the park’s ponds are groundwater dependent, and aquifer overexploitation outside the park is leading to shorter hydroperiods and even desiccation in the pond network. This problem has been exacerbated by the scarcity of rain over the last decade. In 2021-2022, we surveyed the occurrence of the park’s 11 amphibian species, with a view to comparing our findings with those from the last previous survey in 2003-2004. We mapped the species occurrence within the park for the two survey periods. While all 11 amphibian species could still be found in Doñana, their occurrence had decreased across the board. In 2003-2004, 6 species were present across more than 50% of the sampling area. In contrast, 18 years later, such was the case for only two species (Pelophylax perezi and Hyla meridionalis). Declines were greatest for Epidalea calamita, followed by Pelobates cultripes, Triturus pygmaeus, and Lissotriton boscai. The mean number of species per sampling unit (i.e., grid cell) dropped from 4.3 to 3.1. To preserve Doñana’s amphibian community, it is important to restore the park’s pond network, which implies reducing regional groundwater overexploitation.

Hidden hotspots of amphibian biodiversity in China

Identifying and protecting hotspots of endemism and species richness is crucial for mitigating the global biodiversity crisis. However, our understanding of spatial diversity patterns is far from complete, which severely limits our ability to conserve biodiversity hotspots. Here, we report a comprehensive analysis of amphibian species diversity in China, one of the most species-rich countries on Earth. Our study combines 20 y of field surveys with new molecular analyses of 521 described species and also identifies 100 potential cryptic species. We identify 10 hotspots of amphibian diversity in China, each with exceptional species richness and endemism and with exceptional phylogenetic diversity and phylogenetic endemism (based on a new time-calibrated, species-level phylogeny for Chinese amphibians). These 10 hotspots encompass 59.6% of China's described amphibian species, 49.0% of cryptic species, and 55.6% of species endemic to China. Only four of these 10 hotspots correspond to previously recognized biodiversity hotspots. The six new hotspots include the Nanling Mountains and other mountain ranges in South China. Among the 186 species in the six new hotspots, only 9.7% are well covered by protected areas and most (88.2%) are exposed to high human impacts. Five of the six new hotspots are under very high human pressure and are in urgent need of protection. We also find that patterns of richness in cryptic species are significantly related to those in described species but are not identical.

Historical Demography and Climate Driven Range Shifts in the Blue-spotted Salamander Under the Climate Change Scenarios

This study integrates phylogeography with distributional analysis to understand the demographic history and range dynamics of a limited dispersal capacity amphibian species, Blue-spotted Salamander (Ambystoma laterale), under several climate change scenarios. For this we used an ecological niche modeling approach, together with Bayesian based demographic analysis, to develop inferences regarding this species' demographic history and range dynamics. The current model output was mostly congruent with the present distribution of the Blue-spotted Salamander. However, under both the Last Interglacial and the Last Glacial Maximum bioclimatic conditions, the model predicted a substantially narrower distribution than the present. These predictions showed almost no suitable area in the current distribution range of the species during almost the last 22.000 y before present (ybp). The predictions indicated that the distribution of this species shifted from eastern coast of northern North America to the southern part of the current distribution range of the species. The Bayesian Skyline Plot analysis, which provided good resolution of the effective population size changes over the Blue-spotted Salamander history, was mostly congruent with ecological niche modeling predictions for this species. This study provides the first investigation of the Blue-spotted Salamander’s late-Quaternary history based on ecological niche modeling and Bayesian-based demographic analysis. In terms of the main result of this study, we found that the species' present genetic structure has been substantially affected by past climate changes, and this species has reached current distribution range almost from nothing since the Last Glacial Maximum.

Spatio-temporal analysis of toxigenic genes expression in the growing Bufo gargarizans based on RNA sequencing data

BackgroundBufo gargarizans Cantor, a widely distributed amphibian species in Asia, produces and releases toxins through its retroauricular and granular glands. Although various tissues have been sequenced, the molecular mechanisms underlying the toxin production remain unclear. To elucidate these mechanisms, abdominal skin (non-toxic secretory glands) and retroauricular gland (toxic secreting glands) samples were collected at different time points (3, 6, 12, 24, and 36 months) for RNA sequencing (RNA-seq) and analysis. ResultsIn comparison to the S group during the same period, a total of 3053, 3026, 1516, 1028, and 2061 differentially expressed genes (DEGs) were identified across five developmental stages. Gene Ontology (GO) analysis revealed that DEGs were primarily enriched in biological processes including cellular processes, single-organism processes, metabolic processes, and biological regulation. In terms of cellular components, the DEGs were predominantly localized in the cell and cell parts, whereas molecular function indicated significant enrichment in binding and catalytic activity. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that the metabolism and synthesis of various substances, such as lipid metabolism, cofactor and vitamin metabolism, tryptophan metabolism, steroid biosynthesis, and primary bile acid biosynthesis, were accompanied by the development of toads. Additionally, using trend analysis, we discovered candidate genes that were upregulated in the retroauricular glands during development, and the abundance of these genes in the abdominal skin was extremely low. Finally, we identified 26 genes that are likely to be involved in toxin production and that are likely to be involved in toxin anabolism. ConclusionOverall, these results provide new insights into the genes involved in toxin production in B. gargarizans, which will improve our understanding of the molecular mechanisms underlying toxigenic gene expression.

Small Paa Frog and Marbled Cascade Frog are not endemic to Nepal: a response to Tachamo-Shah et al. 2023

Response to ‘Wetland biodiversity of Ramaroshan Lake complex: a need for conservation’ on JoTT Vol 16 issue 12 <https://doi.org/10.11609/jott.7918.15.12.24299-24320>. Many information in the article promising, highlighting the additional attention the area should receive for conservation. However, there are some aspects, particularly that are related to conservation status and endemism of amphibian species which is misleading, making me to realize to write this response.

Investigations on Xenopus laevis body composition and feeding behavior in a laboratory setting

The African clawed frog, Xenopus laevis, has been used as a laboratory animal for decades in many research areas. However, there is a lack of knowledge about the nutritional physiology of this amphibian species and the feeding regimen is not standardized. The aim of the present study was to get more insights into the nutrient metabolism and feeding behavior of the frogs. In Trial 1, adult female X. laevis were fed either a Xenopus diet or a fish feed. After 4 weeks, they were euthanized, weighed, measured for morphometrics and dissected for organ weights and whole-body nutrient analysis. There were no significant differences between the diet groups regarding the allometric data and nutrient contents. The ovary was the major determinant of body weight. Body fat content increased with body weight as indicator of energy reserves. In Trial 2, 40 adult female frogs were monitored with a specifically developed digital tracking system to generate heat-maps of their activity before and up to 25 min after a meal. Three diets (floating, sinking, floating & sinking) were used. The main feed intake activity was fanning the feed into the mouth, peaking until 20 min after the meal. The different swimming characteristics of the diets thereby influenced the activity of the animals. Our dataset helps to adjust the feeding needs to the physical composition and also to meet the natural behavioral patterns of feed intake as a prerequisite of animal wellbeing and animal welfare in a laboratory setting.

Variations in Genetic Diversity of Invasive Species Lithobates catesbeianus in China.

The introduction and subsequent range expansion of the American bullfrog (Lithobates catesbeianus) is part of a rising trend of troublesome biological invasions happening in China. This detrimental amphibious invasive species has strong adaptability. After its introduction and spread, it established its own ecological niche in many provinces of China, and its range has continued to expand to more areas. Previous studies recorded the introduction time of bullfrogs and calculated the changes in their genetic diversity in China using mitochondria, but the specific introduction route in China is still unknown. Expanding upon previous research, we employed whole-genome scans (utilizing 2b-RAD genomic sequencing) to examine single nucleotide polymorphisms (SNPs) and microsatellites within Lithobates catesbeianus to screen the genomes of these invasive amphibian species from eight Chinese provinces and two U.S. states, including Kansas, where bullfrogs originate. A total of 1,336,475 single nucleotide polymorphic loci and 17 microsatellite loci were used to calculate the genetic diversity of bullfrogs and their migration pathways. Our results suggest that the population in Hunan was the first to be introduced and to spread, and there may have been multiple introductions of subpopulations. Additionally, the genetic diversity of both the SNP and microsatellite loci in the Chinese bullfrog population was lower than that of the US population due to bottleneck effects, but the bullfrogs can adapt and spread rapidly. This study will offer crucial insights for preventing and controlling future introductions into the natural habitats in China. Additionally, it will assist in devising more precise strategies to manage the existing populations and curtail their continued expansion, as well as aim to improve clarity and originality while mitigating plagiarism risk.

Loss of amphibian species alters periphyton communities in montane ponds

Amphibian larvae can affect the structure and functioning of freshwater ecosystems, but their effects have been little explored although amphibian biodiversity is rapidly declining. Given that larvae of different amphibian species belong to different trophic levels, their effects on freshwater communities and processes can be expected to differ, with herbivores likely having direct effects on algae and predators having indirect effects through trophic cascades. We explored this question through a mesocosm experiment conducted in montane ponds, using an anuran and a urodele species affected by emergent diseases. We used different scenarios of reduction and loss of one or both species, and compared them to a control scenario representing a typical amphibian community in the study area composed of four species, with total larval density held constant. Loss of the anuran resulted in lower chlorophyll concentration and algal density, likely due to replacement by more efficient grazers. Loss of the urodele produced similar trends but weaker, possibly due to an increase of invertebrate grazing activity in the absence of this predator. Our study shows how the loss of amphibian species can alter the structure of montane ponds, but also how the mechanisms involved and the intensity of effects differ for different species.

Mammal species biodiversity in a Costa Rican protected tropical rainforest environment

Tropical rainforest deforestation has led to old-growth forest loss and declines in species diversity worldwide. In the 1980’s, Costa Rica began innovative reforestation efforts and currently 52% of Costa Rica is a tropical forest environment. However, the impact of Costa Rica’s reforestation efforts on regaining species biodiversity is less known. The aim of this study was to consider longitudinal changes in mammal biodiversity at La Selva Biological Station (LSBS) following reforestation. Mammal species census data were collected between 1994 and 2017 by 30-60 trained and experienced Citizen Scientists. We calculated the total number of different species sighted per year, their relative abundance, and the Shannon Index to estimate biodiversity. Linear regression was used to consider change over time and ANOVA was used to consider differences in land and water use by mammal species (p<0.05). Results suggest the total number of different species observed declined, their relative abundance declined, and the Shannon index of biodiversity declined (p<0.05). This decline is similar to that reported for avian fauna, terrestrial amphibian, and dung beetle species reported at LSBS. Our study highlights the concern that past deforestation continues to lead to species decline and the ongoing need for tropical forest stewardship.

Reptiles and amphibians from the fragments of seasonal Atlantic Forest in northern Paraná, Brazil

Amphibians and reptiles play vital roles within intricate trophic networks as predators, prey, pollinators, and seed dispersers, contributing to nutrient cycling and bioturbation. Tropical regions, characterized by their conducive climates, harbor substantial reptilian and amphibian diversity, highlighting their importance for biodiversity conservation. This study endeavors to bridge knowledge gaps by presenting a comprehensive inventory of amphibian and reptile species within the northern region of the Paraná State, Brazil. Employing active searches and acoustic monitoring, we documented 28 species across diverse microhabitats, reflecting ecological diversity. Analyses of abundance, distribution, and diversity revealed comparable trends between dry and wet seasons. Notably, a broad range of generalist species was observed, adapted to varying ecological niches. Our findings emphasize the relevance of detailed faunal inventories to comprehend and manage biodiversity, particularly in understudied regions, contributing to effective conservation strategies in Seasonal Atlantic Rainforest ecosystems.

Long-term, landscape-level assessment of aquatic pesticide exposure to identify amphibian ontological traits affecting vulnerability.

Amphibians worldwide are threatened by habitat loss, some of which is driven by a changing climate, as well as exposure to pesticides, among other causes. The timing and duration of the larval development phase vary between species, thereby influencing the relative impacts of stochastic hydroregime conditions as well as potential aquatic pesticide exposure. We describe the stages of breeding through metamorphosis for eight amphibian species, based on optimal hydroregime conditions, and use a model of pesticide fate and exposure representative of central Florida citrus groves to simulate hydrodynamics based on observed weather data over a 54-year period. Using the Pesticide in Water Calculator and Plant Assessment Tool, we estimated daily wetland depth and pyraclostrobin exposure, with label-recommended application quantities. Species' timing and duration of larval development determined the number of years of suitable hydroregime for breeding and the likelihood of exposure to peak aquatic concentrations of pyraclostrobin. Although the timing of pesticide application determined the number of surviving larvae, density-dependent constraints of wetland hydroregime also affected larval survival across species and seasons. Further defining categorical amphibian life history types and habitat requirements supports the development of screening-level assessments by incorporating environmental stochasticity at the appropriate temporal resolution. Subsequent refinement of these screening-level risk assessment strategies to include spatially explicit landscape data along with terrestrial exposure estimates would offer additional insights into species vulnerability to pesticide exposure throughout the life cycle. Computational simulation of ecologically relevant exposure scenarios, such as these, offers a more realistic interpretation of differential agrichemical risk among species based on their phenology and habits and provides a more situation-specific risk assessment perspective for threatened species. Integr Environ Assess Manag 2024;20:1667-1676. Published 2024. This article is a U.S. Government work and is in the public domain in the USA.

Dissimilarity among Species and Higher Taxa of Amphibians in a Hotspot of Biodiversity and Endemism in the Neotropics

The Mexican Republic ranks fifth in the world in terms of amphibian diversity, and within Mexico, the state of Oaxaca has the greatest amphibian richness and endemism. Unfortunately, various factors, such as land use change and global warming, have caused a global crisis that threatens the conservation of this class. In the face of these threats, an analysis of beta diversity provides information that can be applied to conservation strategies, since its study reveals the spatial scaling of diversity loss and clarifies the mechanisms of regional diversity maintenance. In this work, we analyzed the beta diversity at the species and higher taxa level (order, family, subfamily, genus and species) for the amphibians of Oaxaca and their replacement components and the differences in richness for anurans and caudates separately between physiographic subprovinces. Very high beta diversity was recorded, with higher diversity occurring among caudates (0.92) than among anurans (0.84). Species replacement was the component that most contributed to this result, and the subprovinces with substantial environmental differences had the most dissimilar amphibian communities. The results of this study show the need to implement conservation strategies in subprovinces with high amphibian richness and endemism levels, following the example of Sierra Madre de Oaxaca (SMO), where local communities have developed conservation actions in most of the territory.

Amphibians at risk: Effects of climate change in the southwestern North American drylands

Climate change can affect species' geographical range in several ways, leading to species' geographical range expansion and contraction. Amphibians are the most threatened vertebrate class, and deterioration is due, in part, to the loss of suitable habitat. This taxonomic group is especially sensitive because of their biphasic life cycles (aquatic and terrestrial) and microhabitat requirements. Herein, we propose a combined approach, which uses biological and life history information to understand species' sensitivity to climate change. The goals of our study were to (1) identify the contribution of environmental variables to the broad-scale distribution of amphibians in drylands of the southwestern United States and Mexico, (2) describe how the habitat suitability of amphibians can potentially respond to a range of climate change scenarios across drylands, and (3) identify amphibian traits related to increased vulnerability and adaptation to the specific pressures and mechanisms of climate change projected for the dryland areas. Results show that climate strongly influences the geographic distribution of amphibian diversity in drylands and that most amphibians are expected to lose habitat in the upcoming decades. This study also shows that areas with the greatest loss of high habitat suitability are related to temperature increases and precipitation reductions. Our results build a framework that will significantly improve the current state of knowledge on the drylands' biogeography while providing comprehensive information and analyses for decision-makers to conserve and protect endangered and threatened amphibian species and their habitats.

Surgically Treated Hind-Limb Fibromyxochondroma in a Cranwell’s Horned Frog (Ceratophrys cranwelli)

Ceratophrys cranwelli, commonly known as the Cranwell’s horned frog or Pacman frog, is an amphibian species native to South America. This species has gained interest both as a pet among hobbyists and as a subject of scientific inquiry in veterinary medicine. A two-year-old pet male albino Ceratophrys cranwelli was presented exhibiting lethargy, weight loss, and anorexia, persisting for two months. Clinical examination revealed a fracture localized to the proximal third of the left femur accompanied by an osteolytic process. The patient was discharged with a treatment regimen for suspected secondary nutritional hyperparathyroidism and an ulcerative skin lesion. Nevertheless, due to the progressive deterioration of the left thigh, amputation of the affected limb was proposed and performed at the coxofemoral joint. Histopathological analysis of the excised mass revealed an atypical mesenchymal mass consistent with a fibromyxochondroma. The surgical procedure was conducted under anesthesia induced by a combination of alfaxalone, ketamine, medetomidine, and butorphanol, with postoperative treatment consisting of enrofloxacin, meloxicam, and butorphanol. The patient showed good post-surgical recovery, exhibiting normal physiological and behavioral activities. This report highlights the management and the diagnostic challenges of a progressive limb lesion in a Ceratophrys cranwelli, offering insights into potential therapeutic approaches for similar cases in amphibians.

Batrachochytrium dendrobatidis and Hannemania mite’s relationships with Mexican amphibians in disturbed environments

The rapid transformation and pollution of ecosystems have severely impacted biodiversity. Specifically, anthropogenic activities have imposed adverse effects on amphibians, with evidence suggesting that these activities alter parasite and pathogen interactions within their hosts. To investigate these interactions in areas affected by different anthropogenic activities, our study focused on analyzing a pathogen and a parasite known to interact within the amphibian skin (spongy epidermis layer) and both compromising amphibian health: Batrachochytrium dendrobatidis (Bd), a fungus responsible for chytridiomycosis, a disease associated with massive population declines in amphibians and the Hannemania sp. mite in Mexico. Four sampling areas along the Sonora River were selected, representing different human activities: mining, livestock, wastewater discharge, agriculture, and one in an urban zone. We analyzed 135 amphibians across 10 anuran species. Among these, the most abundant species (Lithobates yavapaiensis) exhibited the highest prevalence of both pathogen and parasite (90.1% and 27.3%, respectively) and was significantly associated with the intensity of Bd-infection. The prevalence of Hannemania mites varied significantly across sampling sites as did Bd prevalence and infection load, with the highest Bd load found at the wastewater discharge site. A significant association between the intensity of Bd-infection and both mite abundance and amphibian species was observed when the sampling site was considered. Additionally, sites with Bd-positive individuals and Hannemania parasitism coincide with refractory elements characterized by mechanical or corrosion resistance. The persistence of these elements in the environment, along with the small particle size (<850 nm) found in sediments, poses a potential risk of internalization, bioaccumulation (e.g., Fe, Co, and Ti), and their transfer through the food chain. It is thus essential to consider monitoring environmental and biotic factors that modulate the relationships between parasites, pathogens, and amphibians if we are to propose conservation strategies adapted to disturbed environments.

Functional traits drive tadpole responses to land use in coastal areas

Abstract Land use changes are considered a significant cause of amphibian declines worldwide. However, little is known regarding which land uses have more pervasive impacts and which help to supplement the biodiversity conserved in protected areas. In addition, amphibian species respond to a broad array of geographical constraints, which may blur responses to disturbance, depending on the species' natural history. Here, we explored whether functional diversity patterns of amphibian assemblages are linked to human land use while considering spatial autocorrelation and phylogenetic relatedness among species. We surveyed tadpoles along 500 km in coastal wetlands in southern Brazil and quantified four traits related to tadpole habitat use and predator avoidance. Spatial components influenced functional richness to a greater extent than land use. Functional evenness was higher in ponds surrounded by more exotic reforestation, and to a lesser extent an effect of crop area. Functional divergence was also associated with reforestation areas. Our results showed that some land uses (reforestation with exotic species and temporary crops) have more pervasive impacts on tadpole communities. Such changes in amphibian communities occurred even at low levels of landscape change. Our findings highlight that conversion to human land use requires complementary approaches (i.e., tadpole and adult responses) to provide early warning signals about habitat modification effects.

Optimised PCR assays for detecting elusive waterfowl from environmental DNA.

For many aquatic and semiaquatic mammal, amphibian and fish species, environmental DNA (eDNA) methods are employed to detect species distribution and to monitor their presence, but eDNA is much less employed for avian species. Here, we developed primers for the detection of true geese and swan species using eDNA and optimised a PCR protocol for eDNA. We selected taiga bean goose (Anser fabalis fabalis) as our focal (sub)species and sampled water from lakes, from which the presence of taiga bean goose was visually confirmed. To test, if taiga bean goose DNA could be detected among DNA of other goose species, we similarly sampled eDNA from a zoo pond housing several Anatidae species. We were able to detect taiga bean goose DNA in all but one of the tested lakes, including the zoo pond. The primers developed are not species-specific, but rather specific to the genus Anser, due to the close relatedness of Anser species, which prevented the development of species-specific primers and the use of, for example, quantitative PCR. We also developed eDNA primers for Branta species and Cygnus species and tested these primers using the same samples. Canada goose (B. canadensis) and barnacle goose (B. leucopsis) DNA were only detected in the zoo pond (in which they were present), as the sampled natural lakes fall outside the range of these species. We detected whooper swan (C. cygnus) DNA in three lakes and the zoo pond (in which the species was present). The eDNA method presented here provides a potential means to monitor elusive goose species and to study the co-occurrence of large waterfowl.