2025 Midwest Fish & Wildlife Conference

AUTHORS: Thomas Raad, University of Nebraska Omaha; Michaela Sanford, University of Nebraska Omaha Department of Biology; Daniel Peacock, University of Nebraska Omaha Department of Biology; Shayda Azadmanesh, University of Nebraska Omaha Department of Biology; Jeremy White, University of Nebraska Omaha Department of Biology; Guoqing Lu, University of Nebraska Omaha Department of Biology; Han Li, University of Nebraska Omaha Department of Biology

ABSTRACT: All bat species of greatest conservation need in the Midwest are nocturnal and cryptic. Traditional capture-based methods for detecting bats become challenging as populations experience drastic declines. With recent technological advancements, acoustic monitoring has become a common method for detecting these species. However, similarities in bat calls across species can make it challenging to confirm their presence. To address this challenge, it is important to explore additional species detection methods that complement capture and acoustic techniques. Bat guano samples are abundant and easy to collect in the field and extracting and sequencing host DNA from fecal samples for species identification has proven effective. Additionally, bat DNA has been collected and sequenced from roost surfaces, water, and soil, allowing species detection directly from the environments where they reside. Traditional fecal and environmental DNA sequencing methods often require polymerase chain reaction (PCR) amplification with species-specific primers. However, recent advances in genomic technology have introduced nanopore sequencing, which, through nanopore adaptive sampling algorithms, allows for selective sequencing of specified genomic regions. This method uses a user-defined reference file during sequencing to target DNA sequences of interest, eliminating the need for PCR amplification and enabling the generation of long DNA reads for further assembly. Moreover, it allows for portable, in-field sequencing. Mammalian mitochondrial DNA, with its high copy numbers in cells, is suitable for nanopore adaptive sampling. In this project, we explore the application of nanopore adaptive sampling for bat species identification. We opportunistically collected bat guano samples, verified species through morphological examination during capture, and are currently processing fecal samples in the lab. We will present the success rate of species identification from fecal DNA, as well as from environmental DNA collected at known bat roosts.