Amphibian Diversity Research Articles

First documentation on herpetofauna diversity in Gunung Belumut Amenity Forest, Peninsular Malaysia: Implications for conservation in an Environmentally Sensitive Area (ESA).

Malaysia is blessed with lush tropical rainforests that harbour an exceptional diversity of amphibians and reptiles. However, compared to other animal groups, amphibians and reptiles have received limited attention in research, despite their ecological significance. With amphibians and reptile species having been declining rapidly due to anthropogenic activities, there is a pressing need to conserve these species and their habitats. Environmentally Sensitive Areas (ESAs) are designated regions that are beneficial due to their critical role in providing essential ecosystem services and serving as repositories of biodiversity. Nonetheless, the classification of ESAs in Malaysia lacks biological elements and only focuses on physical attributes. To enhance the current ESA classification framework by integrating biological components, there is an urgent need to obtain information on diversity and habitat in Malaysia. Therefore, the objectives of this study were twofold: to determine the diversity of amphibians and reptiles in Gunung Belumut Amenity Forest and to conduct a comparative analysis between the herpetofauna assemblages in Gunung Belumut with other forest reserves in Peninsular Malaysia. The survey was carried out between March and June 2022, with additional sampling conducted in February 2023. The Visual Encounter Survey (VES) and pitfall trap methods were employed to survey the herpetofauna species, focusing on both aquatic and terrestrial habitats within the study area. A total of 210 individuals representing 38 species of herpetofauna were recorded, comprising 18 amphibian and 20 reptile species. Amongst the observed species, Limnonectesblythii was the most frequently encountered amphibian, with 59 individuals observed, while the dominant reptile species was Cyrtodactylusconsobrinus, represented by eight individuals. This pioneering study serves as a vital baseline documentation of the amphibian and reptile assemblages in Gunung Belumut Amenity Forest. It provides valuable information for identifying extant herpetofauna species, including those of potential conservation concern or rarity. These findings contribute to ongoing conservation efforts dedicated to the preservation of herpetofauna within the region. By understanding the diversity and distribution patterns of amphibians and reptiles in Gunung Belumut, effective conservation strategies can be developed to protect these species and their habitats.

Using multiplatform LiDAR to identify relationships between vegetation structure and the abundance and diversity of woodland reptiles and amphibians

AbstractRemotely sensed measures of vegetation structure have been shown to explain patterns in the occurrence and diversity of several animal taxa, including birds, mammals, and invertebrates. However, very little research in this area has focused on reptiles and amphibians (herpetofauna). Moreover, most remote sensing studies on animal–habitat associations have relied on airborne or satellite data that provide coverage over relatively large areas but may not have the resolution or viewing angle necessary to measure vegetation features at scales that are meaningful to herpetofauna. Here, we combined terrestrial laser scanning (TLS), unmanned aerial vehicle laser scanning (ULS), and fused (FLS) data to provide the first test of whether vegetation structural attributes can help explain variation in herpetofauna abundance, species richness, and diversity across a woodland landscape. We identified relationships between the abundance and diversity of herpetofauna and several vegetation metrics, including canopy height, skewedness, vertical complexity, volume of vegetation, and coarse woody debris. These relationships varied across species, groups, and sensors. ULS models tended to perform as well or better than TLS or FLS models based on the methods we used in this study. In open woodland landscapes, ULS data may have some benefits over TLS data for modeling relationships between herpetofauna and vegetation structure, which we discuss. However, for some species, only TLS data identified significant predictor variables among the LiDAR‐derived structural metrics. While the overall predictive power of models was relatively low (i.e., at most R2 = 0.32 for ULS overall abundance and R2 = 0.32 for abundance at the individual species level [three‐toed skink (Chalcides striatus)]), the ability to identify relationships between specific LiDAR structural metrics and the abundance and diversity of herpetofauna could be useful for understanding their habitat associations and managing reptile and amphibian populations.

The effects of edge influence on the microhabitat, diversity and life-history traits of amphibians in western Ecuador

Abstract Edge effects change biodiversity patterns and ecological processes, particularly in tropical forests. To understand the synergistic impact of multiple edges, this study examines how edge influence (EI) is associated with life-history traits (snout-vent length and body temperature), diversity and microhabitat of amphibians as well as habitat characteristics in a tropical forest in Ecuador. We used EI, a metric that calculates cumulative effects across all nearby edges, in combination with five environmental variables that are part of the amphibians’ microhabitat (temperature, humidity, slope, canopy cover and leaf litter depth) to understand how their biodiversity patterns are impacted. Our results show that most amphibian species tend to be habitat specialists, and many had an affinity for forest edges and warmer habitats. We do not find significant correlations between EI and amphibian life-history traits and diversity. Our findings corroborate previous results that many amphibian species tend to be positively associated with habitat fragmentation and show that this association is likely driven by thermal regulation.

Mountain caves of the central region of Veracruz: A vertebrate biodiversity reservoir in a Neotropical hotspot.

The mountain region of central Veracruz, Mexico hosts a large system of karst and volcanic caves that are unexplored. In particular, the vertebrates that inhabit these subterranean ecosystems are unknown. This study evaluated the diversity of mammals, birds, reptiles, amphibians, and fish in three environments (euphotic, disphotic, and aphotic) of 16 caves of different geological origin (12 karst caves and 4 volcanic caves) distributed along an altitudinal gradient (300-2400 m a.s.l.). We found a richness of 242 vertebrate species (184 birds, 30 mammals, 15 reptiles, 12 amphibians, and 1 fish) and an abundance of a total of 11,323 individuals (4,969 mammals, 6,483 birds, 36 reptiles, 27 amphibians, and 5 fish). The richness of all vertebrate classes was higher in karst than in volcanic caves. Vertebrate diversity was also higher at mid-altitudes between 600-899 m a.s.l. Diversity varied between environments, where bird and reptile richness was higher in the euphotic environment, while mammal and amphibian diversity was higher in the aphotic environment. The similarity in the composition of vertebrate species does not depend on the distance between karstic and volcanic caves. Volcanic and karst caves shared on average up to 70% and 55% of vertebrate species, which indicates that only 30% and 45% of species, respectively, is different in each cave type. Given the vulnerability and fragility of these subterranean ecosystems, as well as the important diversity that they contain, we recommend including the caves of the central region of Veracruz in the conservation agenda of local governments and communities. Community-based conservation can help ensure the presence of vertebrate species in the caves of this region.

Landscape use and Habitat Configuration Effects on Amphibian Diversity in Southern Brazil Wetlands

Landscape use and Habitat Configuration Effects on Amphibian Diversity in Southern Brazil Wetlands

Diversity and Habitat Preferences of Amphibians in Abbottabad, Pakistan

Diversity and Habitat Preferences of Amphibians in Abbottabad, Pakistan

A comparative assessment of 16S ribosomal RNA and Cytochrome C Oxidase Subunit I (COI) Primers for Amphibian DNA Barcoding

Amphibians, a diverse and ecologically important group, are facing global declines due to various factors, including habitat loss and climate change. Accurate species identification is crucial for effective conservation efforts, and DNA barcoding has emerged as a powerful tool in this regard. This study compares the efficacy of two DNA barcoding primer sets, targeting the 16S ribosomal RNA gene and the Cytochrome Oxidase I (COI) gene, for identifying 20 amphibian species. While both primer sets successfully amplified sequences, the 16S rRNA gene region identified all 20 samples, whereas the COI region identified 14. The amplified sequences, approximately 550 base pairs for 16S rRNA and 658 base pairs for COI facilitated precise taxonomic placement within amphibian families using Neighbor-Joining phylogenetic trees. These findings enhance DNA barcoding methodology and aid in understanding amphibian diversity, crucial for effective conservation strategies amidst global declines drives by habitat loss, diseases, and climate change.

Effect of Landscape Composition and Configuration on the Diversity of Amphibians and Reptiles

Effect of Landscape Composition and Configuration on the Diversity of Amphibians and Reptiles

Herpetofauna of forest fragments within the Agata Mining Ventures Incorporated (AMVI), Agusan del Norte, Caraga Region, Philippines

The Philippines holds an extraordinary record of amphibian and reptile diversity and endemism; however, factors like habitat loss and fragmentation that may result from activities related to resource extraction, such as mining, pose a significant threat to their conservation. This study examined the herpetofauna composition in select forest fragments within one of the important mining sites in Caraga, the Agata Mining Ventures Incorporated, between 2018 and 2021. Standardized time searches along the established two 2km transects in two sampling stations (Lawigan and Tinigabasan) were conducted to record reptiles and amphibians accounting for 880 man-hours. A total of 513 individuals representing 12 species of amphibians, and 17 species of reptiles were recorded in the area in both surveys. Of these species, 62% (n = 18 spp.) are Philippine endemic, 35% (n = 10 spp.) native, and 3% (n = 1 spp.) were invasive alien species. Overall, the Shannon-Weiner diversity index is high (H’=2.52) and varied significantly across survey periods (Mann-Whitney U test = 248.5, p=0.007). The species-area relationship analysis showed a strong positive correlation for total herpetofauna (R2 = 0.98), amphibians (R2 = 0.87), and reptiles (R2 = 0.99), and a significant correlation existed between reptiles and forest area (p = 0.05). The result suggests a strong reliance of herpetofaunal richness to forested area. Combined forest clearing and the effects of mining activities currently happening in the area might threaten these environment-sensitive taxa. Preserving these forest fragments within AMVI and allocating buffer zones is highly recommended. Periodic scientific field-based surveys and re-surveys, encompassing not only amphibians and reptiles but also various other species are recommended to monitor species distributions, population trends and impacts of on-going activities. These efforts aim to update data-driven sources supporting the mining area’s rehabilitation program, thereby contributing to societal and environmental benefits.

Diversity, distribution, habitat preferences and community assemblages of Amphibians and Reptiles in the “Cilento, Vallo di Diano e Alburni” National Park (Campania, Southern Italy)

Amphibians and reptiles are important ecological indicators of ecosystem status and play significant ecological roles. However, their populations are declining globally due to factors such as climate and land-use change. To effectively manage these species, it is crucial to study their distribution in protected areas. In this study, we provide new information on the distribution and ecology of amphibians and reptiles in the “Cilento, Vallo di Diano e Alburni” (CVDA) National Park in southern Italy. We used a comprehensive dataset consisting of 2465 records obtained from fieldwork, citizen science, and literature to evaluate species’ rarity, habitat preferences and community assemblage. We investigated whether differences in species richness across different habitat types and elevations resulted from species selection of different environmental conditions. We quantified species probability of observation in different habitat types and estimated the significance of species-habitat associations. Species rarity was assessed by considering geographic range, population abundance, and habitat breadth. Collected data substantially improved knowledge of the distribution of numerous species compared to published data. Our findings are generally consistent with the species’ ecological information available in Italy, but we also found some species’ peculiar ecological aspects that are little-known. The CVDA National Park emerges as an effective protected area that ensures good conservation status of the herpetofauna, also in the case of species showing population and range declines at the national scale.

Discovering the diversity of tadpoles in the mid-north Brazil: morphological and molecular identification, and characterization of the habitat.

Brazil stands out for presenting the highest amphibian anuran diversity in the world. However, taxonomic studies that address characteristic of larval stage of anurans are incipient, representing only 62% of known species. We assess the species diversity of tadpoles from eastern Maranhão state, mid-northern region of Brazil based on morphological and molecular identification (i.e., 16S rRNA gene fragment), and we also provide characteristics of the habitats occupied by each species. We carried out 30 field samplings during 13 months in 16 environments along an ecotonal area, over five cities inside the limits of state of Maranhão, between the Maranhão Babaçu Forest and Cerrado ecoregions. We searched for tadpoles in a variety of water bodies, and the tadpoles that reached the developmental stage between 34 to 40 Gosner were morphologically identified. The tadpoles collected herein represent 26 species belonging to five families. The external morphology enabled the identification of 24 species, while the molecular data recognized 22 unique evolutionary units. The most represented family was Hylidae (Hylinae 11 spp., Phyllomedusinae one spp.) followed by Leptodactylidae (Leptodactylinae seven spp., Leiuperinae three spp.), Microhylidae (Gastrophryninae two spp.), and Bufonidae (two spp.). Our results show that oral morphology was the most important character for identifying tadpoles based on morphology, and the specific 16S rRNA primer was suitable for molecular identification. This study pioneers the use of both morphological and molecular data to identify tadpoles in the state of Maranhão. It also provides, for the first-time, habitat characteristic for the species. Our study reveals a high number of anuran species sampled at the larval stage in the region, identifies species that require further taxonomic and systematic attention, and extends the geographic distribution of six species, three of which represent new occurrences for the state. Our results strengthen the hypothesis that the diversity of amphibians from Maranhão is underestimated and highlight the importance of herpetological inventories in poorly sampled areas, decentralizing the knowledge of biodiversity.

Anthropogenic landscape change and amphibian diversity in tropical montane biodiversity hotspots: insights from satellite remote sensing in the Madagascar highlands

AbstractThe magnitude of anthropogenic landscape change in tropical montane biodiversity hotspots and its relationship with biodiversity is a global issue that remains ‘locked-in’ in the broad narrative of tropical change in Africa. Over a montane biodiversity hotspot of Madagascar highlands (Ankaratra Massif), we conducted analysis on land cover change with Landsat satellite sensor data to identify the magnitude of change (1995–2016) and on the habitat change–amphibian diversity relationship to understand links with biodiversity. The results evidenced that 17.8% of the biodiversity hotspot experienced change in land cover in only 20 years. That pressured the already threatened forests, particularly since 2005. Of the total forest area in 1995 (2062.7 ha), 21.5% was cleared by 2016 (1618.3 ha). Changes in forest cover followed a bidirectional pattern. While in the period 1995–2005, forests expanded at a rate of 2.0% year−1 (from 2062.7 to 2524.8 ha), the area declined between 2005 and 2016 at a rate of − 4.1% year−1, fourfold the rate reported nationally for Madagascar (− 1.1% year−1). Forest-to-shrubland transitions emerged as being of increasing concern to forest integrity. We identified a significant link between habitat change and amphibian diversity, but only for species richness. Counter to expectations, no significant relationship was found between species richness and deforestation rates, and between microendemism rates and any of the habitat change variables. Species richness responded to the spatiotemporal variability in vegetation dynamics represented by the standard deviation of the Normalized Differenced Vegetation Index (NDVI_std). Species richness was strongly negatively related to NDVI_std in the short-term (R2 = 0.91, p = 0.003) and long-term (R2 = 0.69, p = 0.03), increasing where the spatiotemporal variability in NDVI was lower. The magnitude of changes in this biodiversity hotspot suggests that region-specific assessments are necessary in the context of the tropical change narrative in Africa and should consider conservation policies tailored for local conditions. Reducing deforestation and land conversion rates through a management plan codesigned with local communities is urgent. Habitat change appears to impact on amphibian diversity by altering the functional attributes of the habitat and not just by reducing habitat extent. NDVI_std seems a relevant indirect metric for monitoring such change although other biophysical attributes obtained from satellite sensor data should be integrated and explored.

Amphibian diversity across three adjacent ecosystems in Área de Conservación Guanacaste, Costa Rica.

Amphibians are the most threatened species-rich vertebrate group, with species extinctions and population declines occurring globally, even in protected and seemingly pristine habitats. These 'enigmatic declines' are generated by climate change and infectious diseases. However, the consequences of these declines are undocumented as no baseline ecological data exists for most affected areas. Like other neotropical countries, Costa Rica, including Área de Conservación Guanacaste (ACG) in north-western Costa Rica, experienced rapid amphibian population declines and apparent extinctions during the past three decades. To delineate amphibian diversity patterns within ACG, a large-scale comparison of multiple sites and habitats was conducted. Distance and time constrained visual encounter surveys characterised species richness at five sites-Murciélago (dry forest), Santa Rosa (dry forest), Maritza (mid-elevation dry-rain forest intersect), San Gerardo (rainforest) and Cacao (cloud forest). Furthermore, species-richness patterns for Cacao were compared with historic data from 1987-8, before amphibians declined in the area. Rainforests had the highest species richness, with triple the species of their dry forest counterparts. A decline of 45% (20 to 11 species) in amphibian species richness was encountered when comparing historic and contemporary data for Cacao. Conservation efforts sometimes focus on increasing the resilience of protected areas, by increasing their range of ecosystems. In this sense ACG is unique containing many tropical ecosystems compressed in a small geographic space, all protected and recognised as a UNESCO world heritage site. It thus provides an extraordinary platform to understand changes, past and present, and the resilience of tropical ecosystems and assemblages, or lack thereof, to climate change.

Grazing for biodiversity: Assessing the effects of cattle management practices on wetlands and amphibian communities in central Argentina

Argentina is widely recognized as a key player in the production and consumption of beef, occupying a prominent position in the global context. However, there is a lack of comprehensive research on the environmental consequences of beef production, particularly concerning wetlands and aquatic biodiversity. We aim to assess the effects of cattle management practices (grazing types and stocking density) on wetland attributes and associated amphibian diversity in central Argentina. During two amphibian breeding seasons, we surveyed 117 wetland sites in paddocks with different cattle grazing types (continuous grazing, glyphosate-promoted pastures and rotational grazing) and stocking densities (low and high). We analyzed the effects of cattle management practices on wetland attributes, including water quality parameters, nutrient concentrations, wetland morphometry, and vegetation cover. Effects on amphibian communities were explored using richness, abundance, and species occurrence. General Mixed-effects Models revealed significant increases in nutrients (total solid dissolved, total nitrogen and soluble reactive phosphorous) and a significant decrease in vegetation cover in sites with high cattle stocking density. Likewise, we observed increased soluble reactive solids in wetlands associated with rotational grazing and decreased vegetation cover in those wetlands related to glyphosate-promoted pastures. Generalized Mixed-effects Models revealed that amphibian communities were negatively affected mainly by high stocking density, while abundance and species occurrence of hylids (Boana pulchella and Scinax squalirostris) were also affected by glyphosate-promoted pastures and rotational grazing treatments. Our results indicate that different types of grazing at low stocking density favor amphibian diversity and improve wetland water quality, therefore, reduced stocking density may result in improved wetland conditions and more diverse amphibian communities. Our results also suggest that glyphosate-promoted pastures and rotational grazing, although beneficial in terms of pasture management, may have unintended consequences on water quality in wetlands and amphibian communities. This study will contribute to our understanding of how cattle management practices influence wetland ecosystems and aquatic biodiversity in the most important cattle breeding area of South America, providing valuable insights for conservation efforts in cattle ranching landscapes.

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.

Natural area visitors’ willingness to pay for amphibian conservation in a global biodiversity hotspot

Transfer of harmful pathogens from imported pet animals to natural populations of wildlife is a new challenge in protected area management. Understanding visitors’ preferences for, and value they place on, protecting biodiversity from such disturbances is important in developing effective management interventions. By conducting a visitor survey at two natural areas in the Southern Appalachian region of the United States, a global hotspot of amphibian biodiversity, this study examined natural area visitors’ willingness to pay (WTP) to conserve wild amphibian populations under two different assumptions for the distribution of WTP – normal and log-transformed. Results from the retained model assuming a normal distribution for WTP showed that visitors were willing to pay US$10.55 (95% CI US$9.61—US$11.50) per visit in donation to ensure the persistence of healthy amphibian populations. Determinants of individual WTP under both model specifications included the perceived threat to amphibians from pathogen transmission in natural areas they visit, their self-assessed familiarity with general knowledge about amphibians, the level of importance they placed on the benefits to humans from amphibian biodiversity, and annual household income. These findings are useful in understanding the economic value of amphibian diversity to natural area visitors, and in assessing the viability of a visitor-supported mechanism to fund projects mitigating anthropogenic threats to amphibian biodiversity.

Amphibians and reptiles of the Sierra Gorda de Querétaro Biosphere Reserve, Mexico: species richness, conservation status and comparison with other natural protected areas in central Mexico

The Sierra Gorda de Querétaro Biosphere Reserve (SGQBR) is one of the largest natural protected areas in Mexico; however, little is known about the richness and diversity of amphibians and reptiles. We present an updated list of species of both groups, the conservation status of these species, as well as an analysis of their diversity with respect to other protected natural areas (NPAs) in central Mexico. The SGQBR contains 132 herpetofauna species (35 amphibians and 97 reptiles). The richest and most diverse families for amphibians were Hylidae (anurans) and Plethodontidae (caudates), and for reptiles Phrynosomatidae (lizards), Colubridae and Dipsadidae (snakes). The values of taxonomic diversity of the SGQBR were similar to those for the regional pool considering others NPAs. However, it did not achieve the highest values compared to the adjacent Los Mármoles National Park or Sierra Gorda de Guanajuato Biosphere Reserve. There was a high complementarity in the species composition between El Chico National Park and SGQBR for both herpetofauna groups. Although a formal list is presented, it is necessary to carry out a greater number of studies focused on analyzing diversity, considering functional attributes of the species and the richness by vegetation types.

Vernal Pool Amphibian Inventories in the Temperate Forests of Eastern North America: Can Environmental DNA Replace the Traditional Methods?

Amphibian populations have been globally declining since at least 1990. In the temperate forests of eastern North America, vernal pools offer important breeding habitats to many amphibians but are usually not considered for forest management because of their small size and temporary status. The effective monitoring and management of amphibians rely on accurate knowledge of their spatiotemporal distributions, which is often expensive to collect due to the amount of fieldwork required. In this study, we tested whether eDNA metabarcoding could identify the same amphibian communities as the traditional inventory protocols. We collected eDNA samples in twelve vernal pools in the spring of 2019 and identified their communities via metabarcoding. At each pool, three traditional amphibian inventory methods were used: call surveys, trapping, and active search surveys. In total, 13 amphibian species were detected, with most of them being detected using both eDNA and the traditional methods. When comparing the results of eDNA with the traditional methods, we found that species ecology and behavior are key factors of its detectability via a specific method. With its higher taxonomical precision and repeatability, eDNA metabarcoding allows for the inventorying of amphibian species living and reproducing in vernal pools and their vicinity with accuracy. As eDNA metabarcoding is inexpensive compared to the traditional methods, we conclude that eDNA sampling should be considered for integration as a standard monitoring tool, after an initial assessment of amphibian diversity.

Amphibian Diversity: Where everything starts to flood, Cáceres Municipality, North Pantanal, Central-West Brazil

Faunal inventories contribute to our understanding of regional diversity, and are fundamental for policy and decision-making regarding the management and conservation of large natural areas. This study aimed to inventory and compile information on amphibian species occurring in the North Pantanal region, in the municipal limits of Cáceres, Mato Grosso, Brazil. We used three methods to inventory amphibian species: (1) fieldwork, (2) analysis of specimens deposited in scientific collections, and (3) literature reviews. We registered 49 amphibian species in Cáceres. Of them, 48 species belonged to the Anura order and were distributed across eight families and 20 genera, and one species belonged to the Gymnophiona order (Siphonops paulensis). The families Leptodactylidae (20 spp.) and Hylidae (17 spp.) were dominant in terms of richness, accounting for 75.5% of all species found in Cáceres. The remaining families had between four and one species each. The municipality is strongly influenced by non-forested formations (e.g., the Cerrado and Pantanal) and presents a high species richness for a non-forested location in Brazil. Our findings highlight Cáceres as one of the richest areas in amphibian species in the North Pantanal region, expanding our knowledge regarding frog diversity. This study provides a foundation for future conservation strategies and additional assessments of amphibian species in light of potential population declines and other emerging threats.

Detection of environmental DNA from amphibians in Northern Europe applied in citizen science

AbstractMany species of amphibians in Northern Europe are threatened and the local distributions are rarely described in detail. Application of modern molecular methods provides an important supplementary tool for monitoring the distribution and diversity of amphibians. For this purpose, we designed, tested, validated, and optimized 14 species‐specific assays on genomic DNA extracted from tissue samples to use for quantitative polymerase chain reaction (qPCR) setups targeting mitochondrial DNA from amphibians in freshwater samples. The tests confirmed species specificity for all assays. Considering a systematic definition of the limit of detection for each of the assays, the presented qPCR assays are unlikely to return false positive detection from any co‐occurring species in northern Europe. For field validation, the qPCR assays were applied in a large‐scale nationwide citizen science project in which sampling and qPCR analysis was carried out by high school students. Data from the citizen science project returned the expected results when compared to the known regional distribution of the target species and confirmed the presence of nine out of 14 Danish species of amphibians in the collected freshwater samples. Four out of 2550 qPCR test sets carried out by the high school students required a professional reanalysis in multiple replicates due to initial unexpected results. This emphasizes that efforts from citizen science may generate large amounts of valuable data, as long as the results are carefully scrutinized by experts.