White-nose syndrome (WNS) has reduced the abundance of many bat species within the United States' mid-Atlantic region, including the endangered Indiana (Myotis sodalis), threatened Myotis septentrionalis (northern long-eared bat) and other bats of conservation concern, such as Perimyotis subflavus (tri-colored bat), Myotis leibii (eastern small-footed bat) and Myotis lucifugus (little brown bat). Impacts to Eptesicus fuscus (big brown bat) populations have been negative, positive or neutral. Migratory bat species such as Lasiurus borealis (eastern red bat) are not impacted by WNS. To determine changes within the National Park Service National Capital Region bat communities, I surveyed the area with mist netting and active acoustic sampling (2016–2018) and compared findings to pre-WNS (2003–2004) data. I documented a significant reduction in the numbers and distributions of M. lucifugus and P. subflavus, a decrease in the distribution of M. septentrionalis, and an increase in Eptesicus fuscus. Documented M. septentrionalis reproduction suggests that portions of the National Capital Region may be important bat conservation areas. To explore the relationship between bat life history and passive acoustic sampling echolocation call data, as well as to review the sampling effort required to detect the presence of extant species, I used mist-netting captures and acoustic call data collected 2015 – 2018 within the District of Columbia, Maryland, Pennsylvania, Virginia, and West Virginia. Based on locally estimated scatterplot smoothing line calculations, I determined that the highest levels of maternity season acoustic activity for Eptesicus fuscus and Lasiurus borealis corresponded primarily to lactation periods. To determine sampling effort, I developed simulations based on species accumulation curves for individual species within different physiographic regions and land-cover types. I determined that the smallest sampling efforts typically corresponded to sampling additional sites versus sampling the same sites with more nights. Detection effort varied greatly by species, physiographic region and land-cover type. Eptesicus fuscus diet has been well-studied throughout North America with visual identification methods from fecal and stomach content samples. Next-generation sequencing provides large genetic data sets analyses in a cost-effective manner and has been used to identify bat prey items. I collected Eptesicus fuscus fecal samples from mid-Atlantic regions and used next-generation sequencing to identify their prey. I documented high variation between survey areas, but did not note a clear pattern of urbanization or fragmentation impacts upon Insecta diversity. All order-level taxa that I documented had been previously documented in morphological studies; however, I did document new families, genera and species. However, I would suggest caution in using next-generation sequencing technologies as authoritative sources for documenting new diet taxa, as I noted frequent occurrences of confounding environmental DNA within the samples. / Doctor of Philosophy / White-nose syndrome (WNS) has reduced the abundance of many bat species within the United States' mid-Atlantic region, including the endangered Indiana (Myotis sodalis), threatened Myotis septentrionalis (northern long-eared bat) and other bats of conservation concern, such as the Perimyotis subflavus (tri-colored bat), Myotis leibii (eastern small-footed bat) and Myotis lucifugus (little brown bat). WNS-impacts to Eptesicus fuscus (big brown bat) populations have been negative, positive or neutral. Migratory bat species such as Lasiurus borealis (eastern red bat) are not impacted by WNS. To determine changes within the National Park Service National Capital Region bat communities, I captured bats with mist-netting techniques and recorded bat echolocation calls with active acoustics (20-minute periods; 2016–2018) and compared findings to pre-WNS (2003–2004) data. I documented a significant reduction in the numbers and distributions of M. lucifugus and P. subflavus, a decrease in the distribution of M. septentrionalis, and an increase in Eptesicus fuscus. Documented M. septentrionalis reproduction suggests that portions of the National Capital Region may be important bat conservation areas. To explore the relationship between bat echolocation calls recorded with passive (over-night) acoustic data and bat reproduction trends, as well as the amount of effort required to document bat species, I sampled 849 sites with passive acoustics and 482 sites with mist-netting during 2016 – 2018 within the District of Columbia, Maryland, Pennsylvania, Virginia, and West Virginia. I determined that the highest levels of acoustic activity for Eptesicus fuscus and Lasiurus borealis (eastern red bat) within the maternity season are associated with the period of highest levels of lactation. To determine sampling effort, I used simulations to detect species in different landscape conditions. I determined that sampling additional sites requires less sampling effort than sampling more nights. The amount of effort required to detect each bat species varied, as did the impact of different landscape conditions. Eptesicus fuscus diet has been well-studied throughout North America by identifying insect parts in feces and stomach contents. Next-generation sequencing is a process which produced large amounts of genetic data sets in a cost-effective manner, and it can be used to identify prey within bat feces. I collected Eptesicus fuscus fecal samples from mid-Atlantic regions and used next-generation sequencing to identify potential prey. I determined that bat diet varied greatly based on survey area and number of feces analyzed. At the order-level, I did not document new taxa: however, I did document new families, genera and species. However, I would suggest caution in using next-generation sequencing technologies as authoritative sources for documenting new diet taxa, as many taxa apparent within my samples were subsequently excluded, as the bats may have been incidentally ingested them.