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Identification of evolutionary hotspots based on genetic data from multiple terrestrial and aquatic taxa and gap analysis of hotspots in protected lands encompassed by the South Atlantic Landscape Conservation Cooperative.

The southeastern United States is a recognized hotspot of biodiversity for a variety of aquatic taxa, including fish, amphibians, and mollusks. Unfortunately, the great diversity of the area is accompanied by a large proportion of species at risk of extinction . Gap analysis was employed to assess the representation of evolutionary hotspots in protected lands w h ere an evolutionary hotspot was defined as an area with high evolutionary potential and measured by atypical patterns of genetic divergence, genetic diversity, and to a lesser extent genetic similarity across multiple terrestrial or aquatic taxa. A survey of the primary literature produced 16 terrestrial and 14 aquatic genetic datasets for estimation of genetic divergence and diversity. Relative genetic diversity and divergence values for each terrestrial and aquatic dataset were used for interpolation of multispecies genetic surfaces and subsequent visualization using ArcGIS. The multispecies surfaces interpolated from relative divergences and diversity data identified numerous evolutionary hotspots for both terrestrial and aquatic taxa , many of which were afforded some current protection. For instance, 14% of the cells identified as hotspots of aquatic diversity were encompassed by currently protected areas. Additionally, 25% of the highest 1% of terrestrial diversity cells were afforded some level of protection. In contrast, areas of high and low divergence among species, and areas of high variance in diversity were poorly represented in the protected lands. Of particular interest were two areas that were consistently identified by several different measures as important from a conservation perspective. These included an area encompassing the panhandle of Florida and southern Georgia near the Apalachicola National Forest (displaying varying levels of genetic divergence and greater than average levels of genetic diversity) and a large portion of the coastal regions of North and South Carolina (displaying low genetic divergence and greater than average levels of genetic diversity) . Our results show the utility o f genetic data sets for identifying cross - species patterns of genetic diversity and divergence (i.e., evolutionary hotspots) in aquatic and terrestrial environments for use in conservation design and delivery across the southeastern United States.

Alabama, Florida, Georgia, North Carolina, South C

Generic reclassification and species boundaries in the rediscovered freshwater mussel ‘Quadrula’ mitchelli (Simpson in Dall, 1896)

The Central Texas endemic freshwater mussel, Quadrula mitchelli (Simpson in Dall, 1896), had been presumed extinct until relict populations were recently rediscovered. To help guide ongoing and future conservation efforts focused on Q . mitchelli we set out to resolve several uncertainties regarding its evolutionary history, specifically its unknown generic position and untested species boundaries. We designed a molecular matrix consisting of two loci ( cytochrome c oxidase subunit I and internal transcribed spacer I ) and 57 terminal taxa to test the generic position of Q . mitchelli using Bayesian inference and maximum likelihood phylogenetic reconstruction. We also employed two Bayesian species validation methods to test five a priori species models (i.e. hypotheses of species delimitation). Our study is the first to test the generic position of Q. mitchelli and we found robust support for its inclusion in the genus Fusconaia. Accordingly, we introduce the binomial, Fusconaia mitchelli comb. nov., to accurately represent the systematic position of the species. We resolved F. mitchelli individuals in two well supported and divergent clades that were generally distinguished as distinct species using Bayesian species validation methods, although alternative hypotheses of species delineation were also supported. Despite strong evidence of genetic isolation within F. mitchelli , we do not advocate for species-level status of the two clades as they are allopatrically distributed and no morphological, behavioral, or ecological characters are known to distinguish them. These results are discussed in the context of the systematics, distribution, and conservation of F. mitchelli .

Conservation Genetics

Integrative taxonomy resolves taxonomic uncertainty for freshwater mussels being considered for protection under the U.S. Endangered Species Act

Objectively delimiting species boundaries remains an important challenge in systematics and becomes urgent when unresolved taxonomy complicates conservation and recovery efforts. We examined species boundaries in the imperiled freshwater mussel genus Cyclonaias (Bivalvia: Unionidae) using morphometrics, molecular phylogenetics, and multispecies coalescent models to help guide pending conservation assessments and legislative decisions. Congruence across multiple lines of evidence indicated that current taxonomy overestimates diversity in the C. pustulosa species complex. The only genetically and morphologically diagnosable species in the C. pustulosa species complex were C. pustulosa and C. succissa and we consider C. aurea , C. houstonensis, C. mortoni , and C. refulgens to be synonyms of C. pustulosa . In contrast, all three species in the C. nodulata complex ( C. necki , C. nodulata , and C. petrina ) were genetically, geographically, and morphologically diagnosable. Our findings have important conservation and management implications, as three nominal species ( C. aurea , C. houstonensis , and C. petrina ) are being considered for protection under the Endangered Species Act.

Scientific Reports

Genetic and morphological characterization of the freshwater mussel clubshell species complex (Pleurobema clava and Pleurobema oviforme) to inform conservation planning

The shell morphologies of the freshwater mussel species Pleurobema clava (federally endangered) and Pleurobema oviforme (species of concern) are similar, causing considerable taxonomic confusion between the two species over the last 100 years. While P. clava was historically widespread throughout the Ohio River basin and tributaries to the lower Laurentian Great Lakes, P. oviforme was confined to the Tennessee and the upper Cumberland River basins. We used two mitochondrial DNA (mtDNA) genes, 13 novel nuclear DNA microsatellite markers, and shell morphometrics to help resolve this taxonomic confusion. Evidence for a single species was apparent in phylogenetic analyses of each mtDNA gene, revealing monophyletic relationships with minimal differentiation and shared haplotypes. Analyses of microsatellites showed significant genetic structuring, with four main genetic clusters detected, respectively, in the upper Ohio River basin, the lower Ohio River and Great Lakes, and upper Tennessee River basin, and a fourth genetic cluster, which included geographically intermediate populations in the Ohio and Tennessee river basins. While principal components analysis (PCA) of morphometric variables (i.e., length, height, width, and weight) showed significant differences in shell shape, only 3% of the variance in shell shape was explained by nominal species. Using Linear Discriminant and Random Forest (RF) analyses, correct classification rates for the two species' shell forms were 65.5% and 83.2%, respectively. Random Forest classification rates for some populations were higher; for example, for North Fork Holston (HOLS), it was >90%. While nuclear DNA and shell morphology indicate that the HOLS population is strongly differentiated, perhaps indicative of cryptic biodiversity, we consider the presence of a single widespread species the most likely biological scenario for many of the investigated populations based on our mtDNA dataset. However, additional sampling of P. oviforme populations at nuclear loci is needed to corroborate this finding.

Indiana, Kentucky, Ohio, Pennsylvania, Tennessee,

Gene pool boundaries for the Yosemite toad (Anaxyrus canorus) reveal asymmetrical migration within meadow neighborhoods

The Yosemite toad ( Anaxyrus [ Bufo ] canorus ) is a federally threatened species of meadow-specializing amphibian endemic to the high-elevation Sierra Nevada Mountains of California. The species is one of the first amphibians to undergo a large demographic collapse that was well-documented, and is reputed to remain in low abundance throughout its range. Recent phylogeographic work has demonstrated that Pleistocene toad lineages diverged and then admixed to differing extents across an elevational gradient. Although lineage divisions may have significant effects on evolutionary trajectories over large spatial and temporal scales, present-day population dynamics must be delineated in order to manage and conserve the species effectively. In this study, we used a double-digest RADseq dataset to address three primary questions: (1) Are single meadows or neighborhoods of nearby meadows most correlated with population boundaries? (2) Does asymmetrical migration occur among neighborhoods of nearby meadows? (3) What topographic or hydrological variables predict such asymmetrical migration in these meadow neighborhoods? Hierarchical STRUCTURE and AMOVA analyses suggested that populations are typically circumscribed by a single meadow, although 84% of meadows exist in neighborhoods of at least two meadows connected by low levels of migration, and over half (53%) of neighborhoods examined display strong asymmetrical migration. Meadow neighborhoods often contain one or more large and flat “hub” meadows that experience net immigration, surrounded by smaller and topographically rugged “satellite” meadows with net emigration. Hubs tend to contain more genetic diversity and could be prioritized for conservation and habitat management and as potential sources for reestablishment efforts.

California

Use of non-invasive genetics to generate core-area population estimates of a threatened predator in the Superior National Forest, USA

Canada lynx (Lynx canadensis) are found in boreal forests of Canada and Alaska and range southward into the contiguous United States. Much less is understood about lynx in their southern range compared to northern populations. Because lynx are currently listed as threatened under the US Endangered Species Act but have recently been recommended for delisting, information on their southern populations is important for lynx recovery, conservation, and management. We used non-invasive, genetic data collected during lynx snowtracking surveys from 2012-2017 to generate core-area estimates of abundance, trend and density in selected core areas of the Superior National Forest of Minnesota, USA. Lynx abundance estimates averaged 41.8 (SD=14.7, range=24-67) during 2012-2017 in the smaller core areas and averaged 52.3 (SD=8.3, range=43-59) during 2015-2017 in the larger core areas. We found no evidence for a decrease or increase in abundance during either period. Lynx density estimates were approximately 7-10 times lower than densities of lynx in northern populations at the low of the snowshoe hare (Lepus americanus) population cycle. To our knowledge, our results are the first attempt to estimate abundance, trend and density of lynx in Minnesota using non-invasive genetic capture-mark-recapture. Estimates such as ours provide useful benchmarks for future comparisons by providing a context with which to assess 1) potential changes in forest management that may affect lynx recovery and conservation, and 2) possible effects of climate change on the depth, density, and duration of annual snow cover and correspondingly, potential effects on snowshoe hares as well.

Minnesota

The needs of salmon and steelhead in balancing their conservation and use

Over the past 100 years, Pacific salmon Oncorhynchus spp. and steelhead trout O. mykiss populations in the Pacific Northwest have experienced dramatic declines as a result of human population growth and associated development of the region's natural resources. Any strategy to reverse those declines will depend on achieving consensus among a diverse group of stakeholders willing to (1) restore damaged habitats and watersheds; (2) consider the biological needs of salmonid fishes; and (3) understand, conserve, and manage existing levels of biodiversity within an ecosystem context. The objective of this chapter is to review and summarize the needs of Pacific salmon and steelhead trout from perspectives that promote their sustainability and perpetuity, and, within that context, provide a framework, for the development of a sustainable fisheries strategy. Although volumes of information are available on the life histories and habitat requirements of Pacific salmonids, new concepts have begun to address the importance of habitat complexity, genetic diversity among locally adapted populations, and the need for appropriate units of conservation. Habitat variability and complexity are the templates that produce diverse, locally adapted populations of Pacific salmon and steelhead trout. Molded by conditions in the environments they colonized, salmon and steelhead have unique adaptations they may have taken hundred i?? perhaps thousands i?? of years to evolve. Any strategy to reverse the declines of salmon and steelhead must focus on conserving both current levels of genetic diversity as well as restoration and maintenance of habitats compatible with the specific needs of individual populations. An argument can be made that we know enough already about the needs of our salmon and steelhead to allow us to make appropriate decisions. There is strong need to integrate what we know about local salmonid adaptations and habitat needs with conservation and management strategies that address the problems we have created. It may be possible to have healthy, sustainable salmonid populations coastwide if we do not wait too long to make up our minds.

Book chapter

Data mining reveals tissue-specific expression and host lineage-associated forms of Apis mellifera filamentous virus

Apis mellifera filamentous virus (AmFV) is a large double-stranded DNA virus of uncertain phylogenetic position that infects honey bees ( Apis mellifera ). Little is known about AmFV evolution or molecular aspects of infection. Accurate annotation of open-reading frames (ORFs) is challenged by weak homology to other known viruses. This study was undertaken to evaluate ORFs (including coding-frame conservation, codon bias, and purifying selection), quantify genetic variation within AmFV, identify host characteristics that covary with infection rate, and examine viral expression patterns in different tissues.

PeerJ

Status of greater sage-grouse in the Bi-State Distinct Population Segment—An evaluation of population trends, habitat selection, and efficacy of conservation actions

Preface This study was completed to provide timely scientific information regarding greater sage-grouse population trends, habitat selection, and the efficacy of previous conservation actions implemented to benefit the Bi-State Distinct Population Segment (DPS). Specifically, we provide these analyses to inform the current (2024) status review and pending listing decision for the DPS being undertaken by the U.S. Fish and Wildlife Service. These findings provide updated, detailed, and comprehensive information regarding the status of a geographically isolated and genetically distinct population of a species of high conservation concern and their habitat. Importantly, this report also provides information on the efficacy of previously implemented conservation actions targeting the Bi-State DPS in a framework that is transferable throughout the species’ range.

California, Nevada

Northwest Boreal Landscape Conservation Cooperative strategic plan 2015 - 2025

The Northwest Boreal Landscape Conservation Cooperative (NWB LCC) is a voluntary, diverse, self-directed management-science partnership, informing and promoting integrated science, sustainable natural and cultural resource management, and conservation to address impacts of climate change and other stressors within and across ecosystems. The NWB LCC area includes parts of Alaska, Yukon, Northwest Territories, and British Columbia. Our partnership reflects both the broad geographic scope and an extensive array of active and engaged participants including resource management organizations, government representatives, policy makers, Tribes and First Nations, industry leaders, researchers, non-governmental organizations, and research/education institutions. Bringing together diverse partners will help assure the northwest boreal is a functioning, sustainable landscape. We live in an era of profound conservation challenges, including the loss and fragmentation of habitats, genetic isolation, invasive species, and unnatural wildfire. The effects of rapidly changing climate are already evident on the landscape. In these circumstances, it is imperative that natural resource management agencies, science providers, Tribes, First Nations, conservation organizations, and other stakeholders work together to understand the drivers and impacts of landscape change and to determine how best to address those challenges. Further, it is essential that the public and communities receive clear communication about the vision and activities of the NWB LCC. Open public access to NWB LCC activities and products will promote acceptance and support of the science that guides potential changes in management action and conservation strategy. This strategic plan provides a great opportunity for the NWB LCC to share our approach and intentions to the LCC members, collaborators, communities, and the public at large.

Report

A genetic assessment of natural barriers for isolating a habitat network proposed for Greenback Cutthroat Trout reintroduction

Objective: Native inland trout conservation efforts rely on physical barriers to exclude nonnative salmonids from target habitats. We used genetic techniques to evaluate a series of natural waterfalls for their potential to serve as barriers to prevent nonnative salmonids from entering a proposed reintroduction area for federally threatened Greenback Cutthroat Trout Oncorhynchus virginalis stomias . Methods: Genetic samples were collected from nonnative Brook Trout Salvelinus fontinalis at 11 sampling reaches above and below natural waterfalls (height: ~1–3 m under base flow conditions) along a 33-km segment of Colorado's upper Cache la Poudre River near the outflow of the proposed reintroduction area. To evaluate whether upstream movement of Brook Trout is restricted by any of these waterfalls, we characterized longitudinal trends in genetic diversity along the river corridor and examined patterns of genetic differentiation and population structure in relation to waterfall locations using a panel of microsatellites. Result: We found no evidence that the waterfalls served as complete movement barriers for nonnative Brook Trout based on genetic clustering analyses, estimates of population differentiation, and longitudinal genetic patterns. Our multilocus assessment did not identify alleles restricted to downstream reaches, and the river segment was genetically homogenized. Conclusion: Our evaluation suggests that the existing waterfalls do not fully prevent upstream movement by nonnative Brook Trout, and thus barrier modification would be needed to establish an isolated Greenback Cutthroat Trout population in the proposed wilderness area.

Colorado

A genetic warning system for a hierarchically structured wildlife monitoring framework

Genetic variation is a well-known indicator of population fitness yet is not typically included in monitoring programs for sensitive species. Additionally, most programs monitor populations at one scale, which can lead to potential mismatches with ecological processes critical to species' conservation. Recently developed methods generating hierarchically nested population units (i.e., clusters of varying scales) for greater sage-grouse ( Centrocercus urophasianus ) have identified population trend declines across spatiotemporal scales to help managers target areas for conservation. The same clusters used as a proxy for spatial scale can alert managers to local units (i.e., neighborhood-scale) with low genetic diversity, further facilitating identification of management targets. We developed a genetic warning system utilizing previously developed hierarchical population units to identify management-relevant areas with low genetic diversity within the greater sage-grouse range. Within this warning system we characterized conservation concern thresholds based on values of genetic diversity and developed a statistical model for microsatellite data to robustly estimate these values for hierarchically nested populations. We found that 41 of 224 neighborhood-scale clusters had low genetic diversity, 23 of which were coupled with documented local population trend decline. We also found evidence of cross-scale low genetic diversity in the small and isolated Washington population, unlikely to be reversed through typical local management actions alone. The combination of low genetic diversity and a declining population suggests relatively high conservation concern. Our findings could further facilitate conservation action prioritization in combination with population trend assessments and (or) local information, and act as a base-line of genetic diversity for future comparison. Importantly, the approach we used is broadly applicable across taxa.

Ecological Applications

A rapid, strong, and convergent genetic response to urban habitat fragmentation in four divergent and widespread vertebrates

Background: Urbanization is a major cause of habitat fragmentation worldwide. Ecological and conservation theory predicts many potential impacts of habitat fragmentation on natural populations, including genetic impacts. Habitat fragmentation by urbanization causes populations of animals and plants to be isolated in patches of suitable habitat that are surrounded by non-native vegetation or severely altered vegetation, asphalt, concrete, and human structures. This can lead to genetic divergence between patches and in turn to decreased genetic diversity within patches through genetic drift and inbreeding. Methodology/Principal Findings: We examined population genetic patterns using microsatellites in four common vertebrate species, three lizards and one bird, in highly fragmented urban southern California. Despite significant phylogenetic, ecological, and mobility differences between these species, all four showed similar and significant reductions in gene flow over relatively short geographic and temporal scales. For all four species, the greatest genetic divergence was found where development was oldest and most intensive. All four animals also showed significant reduction in gene flow associated with intervening roads and freeways, the degree of patch isolation, and the time since isolation. Conclusions/Significance: Despite wide acceptance of the idea in principle, evidence of significant population genetic changes associated with fragmentation at small spatial and temporal scales has been rare, even in smaller terrestrial vertebrates, and especially for birds. Given the striking pattern of similar and rapid effects across four common and widespread species, including a volant bird, intense urbanization may represent the most severe form of fragmentation, with minimal effective movement through the urban matrix.

California

Genome-wide SNP data and morphology support the distinction of two new species of Kovarikia Soleglad, Fet & Graham, 2014 endemic to California (Scorpiones, Vaejovidae)

Morphologically conserved taxa such as scorpions represent a challenge to delimit. We recently discovered populations of scorpions in the genus Kovarikia Soleglad, Fet & Graham, 2014 on two isolated mountain ranges in southern California. We generated genome-wide single nucleotide polymorphism data and used Bayes factors species delimitation to compare alternative species delimitation scenarios which variously placed scorpions from the two localities with geographically adjacent species or into separate lineages. We also estimated a time-calibrated phylogeny of Kovarikia and examined and compared the morphology of preserved specimens from across its distribution. Genetic results strongly support the distinction of two new lineages, which we describe and name here. Morphology among the species of Kovarikia was relatively conserved, despite deep genetic divergences, consistent with recent studies of stenotopic scorpions with limited vagility. Phylogeographic structure discovered in several previously described species also suggests additional cryptic species are probably present in the genus.

California

Genetic structure among greater white-fronted goose populations of the Pacific Flyway

An understanding of the genetic structure of populations in the wild is essential for long-term conservation and stewardship in the face of environmental change. Knowledge of the present-day distribution of genetic lineages (phylogeography) of a species is especially important for organisms that are exploited or utilize habitats that may be jeopardized by human intervention, including climate change. Here, we describe mitochondrial (mtDNA) and nuclear genetic (microsatellite) diversity among three populations of a migratory bird, the greater white-fronted goose ( Anser albifrons ), which breeds discontinuously in western and southwestern Alaska and winters in the Pacific Flyway of North America. Significant genetic structure was evident at both marker types. All three populations were differentiated for mtDNA, whereas microsatellite analysis only differentiated geese from the Cook Inlet Basin. In sexual reproducing species, nonrandom mate selection, when occurring in concert with fine-scale resource partitioning, can lead to phenotypic and genetic divergence as we observed in our study. If mate selection does not occur at the time of reproduction, which is not uncommon in long-lived organisms, then mechanisms influencing the true availability of potential mates may be obscured, and the degree of genetic and phenotypic diversity may appear incongruous with presumed patterns of gene flow. Previous investigations revealed population-specific behavioral, temporal, and spatial mechanisms that likely influence the amount of gene flow measured among greater white-fronted goose populations. The degree of observed genetic structuring aligns well with our current understanding of population differences pertaining to seasonal movements, social structure, pairing behavior, and resource partitioning.

Ecology and Evolution

Museum genomics provide evidence for persistent genetic differentiation in a threatened seabird species in the Western Atlantic

Connectivity among wildlife populations facilitates exchange of genetic material between groups. Changes to historical connectivity patterns resulting from anthropogenic activities can therefore have negative consequences for genetic diversity, particularly for small or isolated populations. DNA obtained from museum specimens can enable direct comparison of temporal changes in connectivity among populations, which can aid in conservation planning and contribute to understanding of population declines. However, museum DNA can be degraded and only available in low quantities, rendering it challenging for use in population genomic analyses. Applications of genomic methodologies such as targeted sequencing address this issue by enabling capture of shared variable sites, increasing quantity and quality of recovered genomic information. We used targeted sequencing of Ultra-conserved Elements (UCEs) to evaluate potential changes in connectivity and genetic diversity of roseate terns ( Sterna dougallii ) with a breeding distribution in the Northwestern Atlantic and the Caribbean. Both populations experienced range contractions and population declines due to anthropogenic activity in the 20 th century, which has the potential to alter historical connectivity regimes. Instead, we found that the two populations were differentiated historically as well as contemporaneously, with little evidence of migration between them for either time period. We also found no evidence for temporal changes in genetic diversity, although these interpretations may have been limited due to sequencing artifacts caused by the degraded nature of the museum samples. Population structuring in migratory seabirds is typically reflective of low rates of divergence and high connectivity among geographically segregated subpopulations. Our contrasting results suggest the potential presence of ecological mechanisms driving population differentiation, and highlight the value of targeted sequencing on DNA derived from museum specimens to uncover long-term patterns of genetic differentiation in wildlife populations.

Integrative and Comparative Biology

Cryptic invasion of Northern Leopard Frogs ( Rana pipiens ) across phylogeographic boundaries and a dilemma for conservation of a declining amphibian

Anthropogenic introduction of species is a major contributor to loss of biodiversity. Translocations within the range of a species are less frequently recognized, but have the potential for negative effects as well. Genetic mixing may lead to loss of local adaptations or further decline through outbreeding depression. These cryptic invasions may be quite difficult to recognize, but genetic tools can be used to recognize and monitor such intraspecific introductions. Conversely, translocations within species can be an important conservation tool to reduce inbreeding depression and replace lost genetic diversity. Thus, cryptic invasions can be either an aid or a hindrance to conservation efforts. We tested for the presence of non-native genotypes and assessed the extent and nature of introgression in populations of Northern Leopard Frog ( Rana pipiens ) in the southwestern US, where populations have declined to a few remnant populations. The most abundant and diverse complex of populations in the region contained a mitochondrial haplotype that was not native to the western US, probably resulting from the introduction of released pets, laboratory animals, or release during fish stocking. These non-native haplotypes were well integrated into a large complex of ponds and lakes, contributing to high genetic diversity in this area. Logistically, the geographic extent of non-native genetic influence within this population precludes eliminating or controlling the non-native component of this population. We recommend assessing the progress and fate of the introgression over time—along with population fitness parameters—to determine whether this introduction is beneficial or detrimental to population persistence. Meanwhile, translocations from nearby locations with similar environmental conditions have the best prospects for avoiding problems with outbreeding depression in other declining populations and will also most effectively preserve regional genetic diversity.

Biological Invasions

Puerto Rico and Florida manatees represent genetically distinct groups

The West Indian manatee ( Trichechus manatus ) populations in Florida ( T. m. latirostris ) and Puerto Rico ( T. m. manatus ) are considered distinct subspecies and are listed together as endangered under the United States Endangered Species Act. Sustained management and conservation efforts for the Florida subspecies have led to the suggested reclassification of the species to a threatened or delisted status. However, the two populations are geographically distant, morphologically distinct, and habitat degradation and boat strikes continue to threaten the Puerto Rico population. Here, 15 microsatellite markers and mitochondrial control region sequences were used to determine the relatedness of the two populations and investigate the genetic diversity and phylogeographic organization of the Puerto Rico population. Highly divergent allele frequencies were identified between Florida and Puerto Rico using microsatellite (F ST = 0.16; R ST = 0.12 (P < 0.001)) and mitochondrial (F ST = 0.66; &Phi; ST = 0.50 (P < 0.001)) DNA. Microsatellite Bayesian cluster analyses detected two populations (K = 2) and no admixture or recent migrants between Florida (q = 0.99) and Puerto Rico (q = 0.98). The microsatellite genetic diversity values in Puerto Rico (H E = 0.45; N A = 3.9), were similar, but lower than those previously identified in Florida (H E = 0.48, N A = 4.8). Within Puerto Rico, the mitochondrial genetic diversity values (&pi; = 0.001; h = 0.49) were slightly lower than those previously reported (&pi; = 0.002; h = 0.54) and strong phylogeographic structure was identified (F ST global = 0.82; &Phi; ST global = 0.78 (P < 0.001)). The genetic division with Florida, low diversity, small population size (N = 250), and distinct threats and habitat emphasize the need for separate protections in Puerto Rico. Conservation efforts including threat mitigation, migration corridors, and protection of subpopulations could lead to improved genetic variation in the endangered Puerto Rico manatee population.

Florida