Integrating Population Genetics With Long-Term Environmental Monitoring to Evaluate and Guide Vernal Pool Creation for Amphibian Conservation.

The decline of biodiversity, particularly among amphibians, is strongly associated with habitat loss and fragmentation. Vernal pools are a critical ecosystem for many pool-breeding amphibians, but they are often overlooked in wetland protection guidelines. Mitigation efforts include vernal pool creation and restoration, but these efforts have varying success in replacing lost functions. This study investigates the success of created vernal pools through long-term environmental monitoring of wood frogs and spotted salamanders (2014-2023) and integrates population genetics to assess the local population health of the wood frog. First, we monitored and compared environmental parameters and reproductive success of indicator species between natural and created pools in a Pennsylvania state park. We then used microsatellite loci to assess within- and between-pool measures of genetic diversity, population structuring, and gene flow for wood frogs. We found two carefully designed created pools positively contributed to local amphibian population persistence by maintaining similar measures of genetic diversity as compared to natural pools. On the other hand, one poorly created pool was genetically distinct and acted as a population sink. Although our findings offer valuable insights, they are based on a limited sample and may not fully represent the broader landscape. However, by integrating genetic information into long-term monitoring datasets, our interdisciplinary approach enhances our understanding of amphibian population dynamics in vernal pool ecosystems. Our findings imply that the most important factors for restoration practitioners to consider when creating or restoring vernal pools are hydroperiod (12-35 weeks), volume (> 50 m3), depth (≥ 30 cm), and surrounding forest land cover (> 60%). These variables are better predictors of indicator species success than pool type (i.e., natural or created). Ultimately, this study emphasizes the need to accompany restoration efforts with long-term monitoring programs that can be used to make adaptive management decisions in an era of extreme environmental change.

Field validation of an eDNA assay for nutria illuminates a role in invasive species management.

Nutria, or coypu (Myocastor coypus), are invasive semi-aquatic rodents present across the United States, Europe, and Asia. Despite successful eradication efforts in certain areas, nutria have resurged in the mid-Atlantic USA, underscoring the need for advanced monitoring tools. Environmental DNA (eDNA) has emerged as a promising technique for species detection and monitoring. Here, an eDNA assay for nutria using qPCR was field-validated in Virginia, USA, showcasing its potential as a tool for post-eradication monitoring. The findings reveal an association between water levels and detection of nutria eDNA, highlighting the importance of water levels in nutria behavior. A painted turtle assay was introduced to confirm nutria absence and demonstrate the potential of passive sampling. The study showcases the sensitivity and efficiency of eDNA assays, emphasizing their value for monitoring and verifying invasive species eradication.

SNAPSHOT USA 2021: A third coordinated national camera trap survey of the United States.

SNAPSHOT USA is a multicontributor, long-term camera trap survey designed to survey mammals across the United States. Participants are recruited through community networks and directly through a website application (https://www.snapshot-usa.org/). The growing Snapshot dataset is useful, for example, for tracking wildlife population responses to land use, land cover, and climate changes across spatial and temporal scales. Here we present the SNAPSHOT USA 2021 dataset, the third national camera trap survey across the US. Data were collected across 109 camera trap arrays and included 1711 camera sites. The total effort equaled 71,519 camera trap nights and resulted in 172,507 sequences of animal observations. Sampling effort varied among camera trap arrays, with a minimum of 126 camera trap nights, a maximum of 3355 nights, a median 546 nights, and a mean 656 ± 431 nights. This third dataset comprises 51 camera trap arrays that were surveyed during 2019, 2020, and 2021, along with 71 camera trap arrays that were surveyed in 2020 and 2021. All raw data and accompanying metadata are stored on Wildlife Insights (https://www.wildlifeinsights.org/), and are publicly available upon acceptance of the data papers. SNAPSHOT USA aims to sample multiple ecoregions in the United States with adequate representation of each ecoregion according to its relative size. Currently, the relative density of camera trap arrays varies by an order of magnitude for the various ecoregions (0.22-5.9 arrays per 100,000 km2), emphasizing the need to increase sampling effort by further recruiting and retaining contributors. There are no copyright restrictions on these data. We request that authors cite this paper when using these data, or a subset of these data, for publication. Any use of trade, firm, or product names is for descriptive purposes only and does not imply endorsement by the US Government.

PREVALENCE OF RANAVIRUS IN SPOTTED SALAMANDER (AMBYSTOMA MACULATUM) LARVAE FROM CREATED VERNAL POOLS IN WEST VIRGINIA, USA.

Ranavirosis is a disease of high concern for amphibians due to widespread documentation of its lethal and sublethal impacts and its high transmission potential across populations and species. We investigated whether spotted salamander (Ambystoma maculatum) ranavirus prevalence and viral load were associated with habitat characteristics, genetic diversity, corticosterone levels, and body size. In 2015 and 2016, we sampled 34 recently created vernal pools in the Monongahela National Forest, West Virginia, USA. We collected tail clippings from 1,128 spotted salamander larvae and waterborne hormone samples from 436 of those larvae, along with eight environmental characteristics of the pools. Over the 2-yr period, we detected ranavirus in 62% of pools, with prevalence ranging from 0% to 63% (mean, 7.68%). Spotted salamander size was positively correlated with ranavirus presence and viral load; however, we did not find associations between ranavirus prevalence or viral load and habitat characteristics, spotted salamander genetic diversity, relatedness, effective number of breeders, or corticosterone levels. The widespread occurrence of ranavirus in the vernal pools illustrates the potential for rapid natural introduction of the pathogen to created wetlands. Managers could consider monitoring local distributions of ranavirus before creation of new vernal pools to guide strategic placement of the wetlands to minimize occurrence and prevalence of this pathogen.

The update and optimization of an eDNA assay to detect the invasive rusty crayfish (Faxonius rusticus).

Environmental DNA (eDNA) is nuclear or mitochondrial DNA shed into the environment, and amplifying this DNA can serve as a reliable, noninvasive way to monitor aquatic systems for the presence of an invasive species. Assays based on the collection of eDNA are becoming increasingly popular, and, when optimized, can aid in effectively and efficiently tracking invasion fronts. We set out to update an eDNA assay to detect the invasive rusty crayfish, Faxonius rusticus. We tested for species specificity compared to other stream crayfish and field tested the assay at sites with known presence (N = 3) and absence (N = 4) in the Juniata River watershed in central Pennsylvania, USA. To maximize sensitivity, we field tested different storage buffers (Longmire's buffer and ethanol), DNA extraction methods (Qiagen's DNEasy and PowerWater kits), and quantitative polymerase chain reaction (qPCR) chemistries (TaqMan and SYBR green). Our assay confirmed the presence data and performed optimally when filter samples were stored in Longmire's buffer, DNA was extracted with DNeasy Blood and Tissue Kit, and TaqMan qPCR chemistry was utilized. With proper sample processing, our assay allows for accurate, noninvasive detection of F. rusticus in streams.

Genetic analyses reveal cryptic introgression in secretive marsh bird populations.

Hybridization is common in bird populations but can be challenging for management, especially if one of the two parent species is of greater conservation concern than the other. King rails (Rallus elegans) and clapper rails (R. crepitans) are two marsh bird species with similar morphologies, behaviors, and overlapping distributions. The two species are found along a salinity gradient with the king rail in freshwater marshes and the clapper in estuarine marshes. However, this separation is not absolute; they are occasionally sympatric, and there are reports of interbreeding. In Virginia, USA, both king and clapper rails are identified by the state as Species of Greater Conservation Need, although clappers are thought to be more abundant and king rails have a higher priority ranking. We used a mitochondrial DNA marker and 13 diagnostic nuclear single nucleotide polymorphisms (SNPs) to identify species, classify the degree of introgression, and explore the evolutionary history of introgression in two putative clapper rail focal populations along a salinity gradient in coastal Virginia. Genetic analyses revealed cryptic introgression with site-specific rates of admixture. We identified a pattern of introgression where clapper rail alleles predominate in brackish marshes. These results suggest clapper rails may be displacing king rails in Virginia coastal waterways, most likely as a result of ecological selection. As introgression can result in various outcomes from outbreeding depression to local adaptation, continued monitoring of these populations would allow further exploration of hybrid fitness and inform conservation management.

Limited influence of local and landscape factors on finescale gene flow in two pond-breeding amphibians.

Dispersal and gene flow within animal populations are influenced by the composition and configuration of the landscape. In this study, we evaluated hypotheses about the impact of natural and anthropogenic factors on genetic differentiation in two amphibian species, the spotted salamander (Ambystoma maculatum) and the wood frog (Lithobates sylvaticus) in a commercial forest in central Maine. We conducted this analysis at two scales: a local level, focused on factors measured at each breeding pond, and a landscape level, focused on factors measured between ponds. We investigated the effects of a number of environmental factors in six categories including Productivity, Physical, Land Composition, Land Configuration, Isolation and Location. Embryos were sampled from 56 spotted salamander breeding ponds and 39 wood frog breeding ponds. We used a hierarchical Bayesian approach in the program GESTE at each breeding pond and a random forest algorithm in conjunction with a network analysis between the ponds. We found overall high genetic connectivity across distances up to 17 km for both species and a limited effect of natural and anthropogenic factors on gene flow. We found the null models best explained patterns of genetic differentiation at a local level and found several factors at the landscape level that weakly influenced gene flow. This research indicates multiscale investigations that incorporate local and landscape factors are valuable for understanding patterns of gene flow. Our findings suggest that dispersal rates in this system are high enough to minimize genetic structuring and that current forestry practices do not significantly impede dispersal.

Characterizing the width of amphibian movements during postbreeding migration.

Habitat linkages can help maintain connectivity of animal populations in developed landscapes. However, the lack of empirical data on the width of lateral movements (i.e., the zigzagging of individuals as they move from one point to point another) makes determining the width of such linkages challenging. We used radiotracking data from wood frogs (Lithobates sylvaticus) and spotted salamanders (Ambystoma maculatum) in a managed forest in Maine (U.S.A.) to characterize movement patterns of populations and thus inform planning for the width of wildlife corridors. For each individual, we calculated the polar coordinates of all locations, estimated the vector sum of the polar coordinates, and measured the distance from each location to the vector sum. By fitting a Gaussian distribution over a histogram of these distances, we created a population-level probability density function and estimated the 50th and 95th percentiles to determine the width of lateral movement as individuals progressed from the pond to upland habitat. For spotted salamanders 50% of lateral movements were ≤13 m wide and 95% of movements were ≤39 m wide. For wood frogs, 50% of lateral movements were ≤17 m wide and 95% of movements were ≤ 51 m wide. For both species, those individuals that traveled the farthest from the pond also displayed the greatest lateral movement. Our results serve as a foundation for spatially explicit conservation planning for pond-breeding amphibians in areas undergoing development. Our technique can also be applied to movement data from other taxa to aid in designing habitat linkages.