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The U.S. Fish and Wildlife Service allows acoustical surveys and automated identification software to determine the presence of the endangered northern long-eared bat (Myotis septentrionalis) and Indiana bat (Myotis sodalis). Analytical software is required to assess presence probability on a site-night basis using a maximum likelihood estimator (MLE) that accounts for interspecific bat misclassification rates. The current standard for occupancy is a returned MLE P-value < 0.05 at the nightly level irrespective of the number of files identified as either northern long-eared bats or Indiana bats. These MLE P-values can vary based on presence of other bat species with similar calls and the relative proportions of all species recorded. Accordingly, there is concern that with few nightly northern long-eared bat or Indiana bat recordings or the presence of large numbers of high frequency bats, false-negative findings from a swamping effect could result. Using data collected in 2020–2021 by the U.S. Fish and Wildlife Service to set nationwide acoustic monitoring guidelines, we examined the relationship of returned software MLE P-values from 4873 site-nights of acoustic detector data relative to nightly counts of northern long-eared bats and Indiana bats, overall counts of other high-frequency bats, and habitat cover type. For both northern long-eared bats and Indiana bats, nights with one or more echolocation pass files identified as either species but above the MLE P-value threshold largely occurred where nightly counts of the target species was <15 and their proportion to the count of high-frequency bat species was low. We followed this analysis with a simulation using a known call library and observed similar patterns. Accordingly, with few nightly echolocation passes, post-hoc visual assessment following automated software identification easily could be undertaken. Evidence of swamping by other high-frequency species causing positive file identification creating false

Southern fox squirrels (Sciurus niger niger) have been declining due to habitat fragmentation, cover type conversion, and fire suppression in the Southeast. A decrease in growing season burns has led to hardwood encroachment and forest mesophication that benefit the competing eastern gray squirrels (S. carolinensis). In the southern Coastal Plain and Piedmont of Virginia, these pattern raises the question of whether gray squirrels are competitively excluding southern fox squirrels in these altered landscapes. From October 2019 to October 2020, we conducted continual camera trapping for southern fox squirrels and gray squirrels on the Big Woods/Piney Grove Complex (BWPGC) and at Fort Barfoot (FB) in the Coastal Plain and lower Piedmont of Virginia, respectively. Both sites are among the few areas that still contain large, intact pine savanna and mixed-pine hardwood forests in southeastern Virginia. We used two-species occupancy modeling to investigate occupancy estimates of southern fox squirrels and possible competition with gray squirrels, based on detection histories collected from camera traps on BWPGC and FB. We then conducted informed single-species occupancy modeling to estimate the necessary level-of effort (LOE) required to determine the probable absence of southern fox squirrels at sampling sites in the region. No fox squirrels were observed at FB. Our top, two species occupancy model showed that gray squirrel occupancy increased with increasing time since last burn. However, southern fox squirrel occupancy, in the absence of gray squirrels, decreased with increasing time since last burn. Gray squirrels typically inhabited hardwood-dominant closed canopy areas whereas southern fox squirrels did so at BWPGC only in the absence of gray squirrels. This suggests that southern fox squirrels are selecting areas on BWPGC based on resource needs and possibly competition with gray squirrels. A single-season occupancy model confirmed that southern fox squirrel occupa

Conservation of bats declining from white-nose syndrome (WNS) impacts requires an understanding of both temporal and landscape-level habitat relationships. Traditionally, much of the research on bat ecology has focused on behavior of summer maternity colonies within species’ distribution cores, including that of the endangered northern long-eared bat (Myotis septentrionalis). To further our knowledge of this species, we evaluated multi-season activity patterns in eastern North Carolina and Virginia, including areas where populations were recently discovered. We used passive acoustic monitoring to assess relative and probable activity of northern long-eared bats from October 2016 to August 2021. Northern long-eared bat relative activity was greatest in areas containing greater proportions of woody wetlands and upland pine-dominated evergreen forests. However, the likelihood of recording northern long-eared bats was associated with smaller proportions of woody wetlands and open water resources. Furthermore, we observed a higher probability of recording northern long-eared bats during non-winter seasons. Probable activity was greatest at temperatures be- tween 10 and 25 C, potentially highlighting an optimal thermoneutral zone for the species regionally. Relative activity of northern long-eared bats on the Coastal Plain of Virginia and North Carolina was primarily driven by cover features, whereas probable activity was driven by a combination of cover features, seasonality, and temperature. Therefore, acoustical surveys for this species may be most effective when targeting woody wetlands adjacent to upland forests, particularly upland pine-dominated evergreen stands, during moderate temperatures of non-winter seasons (1 April–15 November). Moreover, conservation of a diverse mosaic of woody wetlands juxtaposed by upland forests may promote both roosting and overwintering habitat, thereby enhancing overwintering survival, maternity colony establishment, and ultimately

Understanding the relationships of biotic and abiotic factors to seasonal activity at hibernacula is important for the conservation of bats impacted by white-nose syndrome (WNS). Research on the relative and probable activity patterns of the federally endangered northern long-eared bat (Myotis septentrionalis) primarily has focused on summer maternity colonies, whereas surveys at hibernacula have traditionally relied on external capture and internal counts. We used passive acoustic monitoring to assess the relative and probable activity of northern long-eared bats at 13 hibernac- ula in western Virginia, from August 2020 to May 2022. Northern long-eared bats were most active near hibernacula during warmer weeks of the fall swarm and spring emergence, when rainfall was low. Similarly, the probability of northern long-eared bat activity was highest near hibernacula during the spring/summer season. However, unlike relative activity, the likelihood of recording northern long-eared bats was associated with more heteroge- neous, interior forests. Our results suggest that northern long-eared bat activity largely follows the described pre-WNS hibernation phenology of the species. Therefore, acoustical surveys to monitor northern long-eared bat populations at hibernacula should focus on entrances during peak activity periods (mid-April and early September), rather than the nearby landscape. Finally, management to promote resource-rich foraging habitat adjacent to hibernacula for use during swarming and emergence may increase survival during hibernation, fitness for spring migration, and ultimately, improve the reproductive success of northern long-eared bats in western Virginia.

The potential introduction of SARS-CoV-2, the virus responsible for the COVID-19 pandemic, into North American bat populations is of interest to wildlife managers due to recent disease-mediated declines of several species. Populations of little brown bats (Myotis lucifugus) have collapsed due to white-nose syndrome (WNS), a disease caused by the introduction and spread of the fungal pathogen Pseudogymnoascus destructans (Pd). Throughout much of the United States and southern Canada, large colonies of the species routinely established diurnal roosts in anthropogenic structures, thereby creating the potential for direct human contact and cross-species disease transmission. Given recent declines and the potential for further disease impacts, we collected oral swabs from eight little brown bat maternity colonies to assess the presence and prevalence of SARS-CoV-2 by RT-qPCR analysis. Little brown bat colonies in Maryland (n = 1), New Hampshire (n = 1), New Jersey (n = 2), New York (n = 1), Rhode Island (n = 2), and Virginia (n = 1) were taken during May-August, 2022. From 235 assayed individuals, no bat tested positive for SARS-CoV-2. Our results indicate that little brown bats may not contract SARS-CoV-2 or that the virus persists at undetectable levels in populations of the Mid-Atlantic and Northeast during summer months. Nonetheless, continued monitoring and future work addressing other seasons may still be warranted to conclusively determine infection status.

Along the mid-Atlantic coast of the United States, eastern red bats (Lasiurus borealis) are present during fall mating and migration, though little is currently known about most aspects of bat migration. To reveal migration patterns, and understand drivers of over-water flight, we captured and radio-tagged 115 eastern red bats using novel technology, and subsequently tracked and described their movements throughout the region. We compared over-water flight movements to randomly generated patterns using a use-availability framework, and subsequently used a generalized linear mixed effects model to assess the relationship of over-water flight to atmospheric variables. We used hidden Markov models to assess daily activity patterns and site residency. Most bats with long-distance movements traveled in a southwesterly direction, however path vectors were often oriented interior toward the continental landmass rather than along the coastline. We observed that some bats transited wide sections of the Chesapeake and Delaware bays, confirming their ability to travel across large water bodies. This over-water flight typically occurred in the early hours of the night and during favorable flying conditions. If flight over large water bodies is a proxy for over-ocean flight, then collision risk at offshore wind turbines – a major source of migratory bat fatalities – may be linked nightly to warm temperatures that occur early in the fall season. Risk, then, may be somewhat predictable and manageable with mitigation options linking wind-energy operation to weather conditions and seasonality.

Neotoma magister (Allegheny Woodrat) is a nocturnal, emergent rock-habitat specialist (i.e., inhabits rocky outcrops, boulderfields, and caves). Woodrat populations have declined range-wide due to habitat fragmentation, endoparasites, and interspecific competition. We estimated the diel activity curves of Allegheny Woodrats and assessed the effects of habitat type (exposed rock habitat/cave- exterior vs. cave-interior) and season (spring, summer, and fall) on curve shape. We also investigated the effect of 2 granivorous competitors' presence and activity curves (Peromyscus spp. and Tamias striatus [Eastern Chipmunk]) on woodrat activity. Additionally, we investigated whether the presence or absence of Procyon lotor (Raccoon), a primary carrier of Baylisascaris procyonis (Raccoon Roundworm), significantly affects the presence or absence of Allegheny Woodrats. We used remote-detecting cameras to document the diel cycles of Allegheny Woodrats and 2 competitors across 83 sites in western Virginia and 2 sites in West Virginia from 2017 to 2022. For 13,002 recorded events, we detected woodrats at 36 of 85 sites (3778 camera events). We observed a higher proportion of daytime activity by woodrats within cave interiors than cave exteriors. Allegheny Woodrat activity curves differed among seasons, with the greatest differences observed between summer and fall and with similar to 80% activity overlap. These activity curves differed significantly when co- occurring with versus not co-occurring with a competitor. Additionally, Allegheny Woodrats showed an inverse activity rate with Peromyscus spp. Thus, our results suggest that competition avoidance via temporal partitioning occurs between these species. Allegheny Woodrats and Raccoons occurred together more often than expected suggesting the presence of woodrats is currently not reduced by the presence of Raccoons. Our remote-detecting camera data help elucidate relationships of Allegheny Woodrats with presumptive competitors

The gray bat (Myotis grisescens) is a cave-obligate species that has been listed as federally endangered since 1976, following population declines from human disturbance at hibernation and maternity caves. However, with cave protection, most gray bat populations have increased. As part of a project examining bat use of transportation structures as day-roosts, we continuously acoustically monitored 12 riparian sites within the Clinch River Watershed of southwest Virginia from March through November, 2018–2020. We used 15 different landscape and weather-related variables in generalized linear mixed models to determine factors influencing gray bat presence and activity. Seasonal activity patterns were similar among years, but the number of nightly gray bat calls increased with each passing year, consistent with positive population trends observed at winter hibernacula. Year and average nightly temperatures were positively correlated with gray bat activity, as was, unexpectedly, average nightly wind speed. Total nightly precipitation, distance to the nearest hibernaculum in Tennessee, percent forested area within 2 km of a detector, mean elevation within 2 km of a detector, detector type, and amount of urban development within 2 km of a detector were negatively correlated with gray bat activity. Our findings show where and when gray bat presence is likely in southwest Virginia, thereby helping managers avoid negative impacts from activities such as bridge repair or replacement and planning of future monitoring to track population trends.

Achieving a target population size is often the first goal of species restorations. From 2012 to 2014, the Virginia Department of Wildlife Resources released 75 elk (Cervus canadensis) originating from Kentucky into Buchanan County in southwestern Virginia. These individuals were ear tagged with unique numbers upon release with an additional 33 elk tagged within the Virginia Elk Management Zone (VEMZ) from 2019 through early 2022. To assess post-release population size, we conducted visual driving surveys throughout Buchanan County from January through mid-April, 2021 and January through March, 2022, counting elk and noting sex, age class, and tagged individuals when observed. We conducted four surveys annually, each consisting of pooled elk counts from eight driving routes, and calculated a Lincoln-Petersen population estimate with Chapman’s bias correction for each survey, then averaged estimates for each year. The population estimate in Buchanan County was 250 (95% CI: 100–400) elk in 2021 and 303 (155–452) in 2022. Our elk population estimates indicate Virginia is on the trajectory of meeting the first goal in their 2019–2028 elk management plan of achieving a viable elk population.

Hierarchical conservation and management of Indiana bat (Myotis sodalis) habitat may benefit from use of species distribution models. White-nose syndrome has caused additional declines for this endangered bat, requiring use of historical presence locations for habitat-related analyses. We created random forest presence/pseudo-absence models to assess the distribution and availability of Indiana bat habitat across the 670,000-ha Monongahela National Forest (MNF), West Virginia, USA. We collated historical roost and capture locations, both individually and in combination, to examine impacts of various biotic and abiotic predictors on roosting and foraging habitat of Indiana bats. Our final concordance map suggests that Indiana bat habitat was abundant (37.2% of the MNF) but localized, with predicted suitable areas often associated with edges of dry-calcareous forests. We observed significant variation between models, with the capture-only model independently identifying the greatest amount of potential habitat (47.8%). However, 21.9% of all potential Indiana bat habitat was identified by complete inter-model agreement. Our SDM outputs may assist land managers in identifying avoidance areas and new survey sites (i.e., capture and acoustic sampling) to support forest management activities.

Since the spread of white-nose syndrome in North America, several bat species have shown precipitous declines in abundance and distribution. With lower netting detection probabilities for the currently threatened but proposed endangered northern long-eared bat (Myotis septentrionalis) and endangered Indiana bats (Myotis sodalis), determination of presence or absence for regulatory clearance often has shifted to the use of acoustic surveys. However, acoustic surveys are unable to differentiate between non-reproductive individuals versus a maternity colony. We used recorded nightly echolocation pass counts of bat species-specific probabilities with maximum likelihood estimator (MLE) scores to determine thresholds by cover type and reproductive period whereby the potential for northern long-eared bat or Indiana bat maternity colonies occurs. Where nightly MLE P-values were < 0.05) were higher for sites with observed maternity activity for both bat species across forest, forest-field edge, and riparian areas versus sites where no maternity activity was known. For northern long-eared bats, nightly pass counts were highest in the juvenile volancy period (after 15 July) whereas, for Indiana bats, nightly pass counts were highest in the lactation period (16 June to 15 July). Except for edge conditions for northern long-eared bats, a MLE P < 0.05 combined with nightly pass counts above thresholds developed from surveys at known maternity colony sites for both species may indicate potential presence of a maternity colony locally and provide a tool to more efficiently use targeted mist-netting for further determination.

Species distribution models enable resource managers to avoid and mitigate impacts to, or enhance habitat of, target species at the landscape level. Persistent declines of northern long-eared bats (Myotis septentrionalis) due to white-nose syndrome have made acquisition of contemporary data difficult. Therefore, use of legacy data may be necessary for creation of species distribution models. We used historical roost and capture records, both individually and in combination, to assess the distribution and availability of northern long-eared bat habitat across the 670,000-ha Monongahela National Forest (MNF), West Virginia, USA. We created random forest presence/pseudo-absence models to examine influences of various biotic and abiotic predictors on both roosting and foraging presence locations of northern long-eared bats. Predicted northern long-eared bat habitat was abundant (43.1% of the MNF) and widely dispersed. Generally, all models suggested that northern long-eared bat habitat was characterized by interior forests containing linear edge features. We observed only 3.4% spatial overlap of habitat based on complete model agreement, but 38.5% of all habitat areas resulted from agreement between capture-only and combination models. Our models provide important assessments of habitat availability necessary for addressing state and federal conservation requirements on the MNF and adjacent eastern West Virginia mountains.