1 A conservation genetic analysis of the endangered Florida bonneted bat (Eumops floridanus) Valentina Alvarez 1 , Jeff A. Gore 2 , Cortney Pylant 1 , James D. Austin 1 1 Department of Wildlife Ecology and Conservation, University of Florida 2 Florida Fish and Wildlife Research Institute, Florida Fish and Wildlife Conservation Commission The Florida bonneted bat is one of the rarest bat species in North America, and has been designated as federally endangered since 2013. Very little is known about the basic biology of Florida bonneted bats, and this project was initiated to fill some important gaps regarding its social structure, genetic diversity and mating system of bonneted bats located at 12 artificial roosting colonies in Charlotte County, Florida. This location represents the largest known population of the species in Florida. Samples were collected over a 32-month period (April 2014-December 2016), during nine tri-annual trapping sessions by Florida Fish and Wildlife Conservation Commission biologists and volunteers. Wing punches were collected from all unique bats to allow for estimation of genetic diversity and molecular relatedness among bats to answer questions of the species’ mating system. The objective of this paper was to test whether roosts within the WMA display patterns of isolation by distance and to determine if relatedness among females is greater than among males within roosts. Genetic distance calculated from 13 microsatellite loci revealed a pattern of significant isolation by distance among roosts spanning approximately 16 kms. Relatedness within roosts was greater among females within roosts than among males, reflecting a pattern driven by male-biased dispersal. This is in line with the expectation of a polygynous mating system. These results provide an important initial genetic picture of the diversity and mating system of Florida bonneted bats. INTRODUCTION A thorough understanding of a species’ mating system and social patterns is necessary to understand the implications of its social structure (Burland 2001). Understanding social organization is particularly important when dealing with endangered species because different systems predispose species to certain conservation risks (Pritchard 2000). With a proper understanding of social structure and mating behavior, wildlife managers can formulate appropriate management plans and designate conservation units for endangered species. Many species in the order Chiroptera are highly elusive and nocturnal, making kinship assessment through direct observation difficult (Burland 1999). Molecular methods provide an opportunity for a more thorough assessment of kinship (Hughes 1998). The Florida bonneted bat (Eumops floridanus) is a communally roosting bat and most of the known population occurs within artificial roosts in the Babcock- Webb WMA in Charlotte county, FL (Figure 1). These artificial roosts, placed by the Florida Fish and Wildlife Conservation Commission, provide a unique opportunity to conduct kinship assessments of this species. The mating system and social behavior of E. floridanus is unknown and the initial step to understand the species’ natural history is to analyze dispersal, gene flow, and the genetic relatedness of individuals within and between roosts. Endemic to southern Florida, the Florida bonneted bat was recently elevated from subspecies of E. glaucinus to species status (Timm 2004). Fossil evidence suggests that E. floridanus was once much wider-ranging with a higher density of individuals, (Timm 2004) but populations have decreased due to pesticide use and the loss of appropriate roosting sites (IUCN 2016). The IUCN lists the Florida bonneted bat as a Vulnerable species and the Florida Fish and Wildlife Conservation Commission classifies it as an endangered species. In recent years, microsatellites have become the most widely used DNA marker for conducting kinship assessments since they are ubiquitous and highly variable (Queller 1993). Microsatellites are tandem repeats of short nucleotide motifs which possess high polymorphism, are non-coding, and exhibit codominant inheritance. Cross- specific utility of microsatellite markers is low for bat species (Dawson et al. 2004; Naidoo et al. 2013) and before my study, no species-specific microsatellite markers existed for E. floridanus. Thus, I developed novel microsatellite markers for E. floridanus to identify kinship amongst individuals residing in the artificial roosts within Babcock-Webb WMA.