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The Coalition for Conservation Genetics: Working across organizations to build capacity and achieve change in policy and practice

The Coalition for Conservation Genetics (CCG) brings together four eminent organizations with the shared goal of improving the integration of genetic information into conservation policy and practice. We provide a historical context of conservation genetics as a field and reflect on current barriers to conserving genetic diversity, highlighting the need for collaboration across traditional divides, international partnerships, and coordinated advocacy. We then introduce the CCG and illustrate through examples how a coalition approach can leverage complementary expertise and improve the organizational impact at multiple levels. The CCG has proven particularly successful at implementing large synthesis-type projects, training early-career scientists, and advising policy makers. Achievements to date highlight the potential for the CCG to make effective contributions to practical conservation policy and management that no one “parent” organization could achieve on its own. Finally, we reflect on the lessons learned through forming the CCG, and our vision for the future.

Conservation Science and Practice

Comparing modern identification methods for wild bees: Metabarcoding and image-based morphological taxonomic assignment

With the decline of bee populations worldwide, studies determining current wild bee distributions and diversity are increasingly important. Wild bee identification is often completed by experienced taxonomists or by genetic analysis. The current study was designed to compare two methods of identification including: (1) morphological identification by experienced taxonomists using images of field-collected wild bees and (2) genetic analysis of composite bee legs (multiple taxa) using metabarcoding. Bees were collected from conservation grasslands in eastern Iowa in summer 2019 and identified to the lowest taxonomic unit using both methods. Sanger sequencing of individual wild bee legs was used as a positive control for metabarcoding. Morphological identification of bees using images resulted in 36 unique taxa among 22 genera, and >80% of Bombus specimens were identified to species. Metabarcoding was limited to genus-level assignments among 18 genera but resolved some morphologically similar genera. Metabarcoding did not consistently detect all genera in the composite samples, including kleptoparasitic bees. Sanger sequencing showed similar presence or absence detection results as metabarcoding but provided species-level identifications for cryptic species (i.e., Lasioglossum ). Genus-specific detections were more frequent with morphological identification than metabarcoding, but certain genera such as Ceratina and Halictus were identified equally well with metabarcoding and morphology. Genera with proportionately less tissue in a composite sample were less likely to be detected using metabarcoding. Image-based methods were limited by image quality and visible morphological features, while genetic methods were limited by databases, primers, and amplification at target loci. This study shows how an image-based identification method compares with genetic techniques, and how in combination, the methods provide valuable genus- and species-level information for wild bees while preserving tissue for other analyses. These methods could be improved and transferred to a field setting to advance our understanding of wild bee distributions and to expedite conservation research.

PLoS ONE

Genetic diversity and connectivity of moose (Alces americanus americanus) in eastern North America

Genetic diversity is critical to a population’s ability to overcome gradual environment change. Large-bodied wildlife existing in regions with relatively high human population density are vulnerable to isolation-induced genetic drift, population bottlenecks, and loss of genetic diversity. Moose ( Alces americanus americanus ) in eastern North America have a complex history of drastic population changes. Current and potential threats to moose populations in this region could be exacerbated by loss of genetic diversity and connectivity among subpopulations. Existing genetic diversity, gene flow, and population clustering and fragmentation of eastern North American moose are not well quantified, while physical and anthropogenic barriers to population connectivity already exist. Here, single nucleotide polymorphism (SNP) genotyping of 507 moose spanning five northeastern U.S. states and one southeastern Canadian province indicated low diversity, with a high proportion of the genomes sharing identity-by-state, with no consistent evidence of non-random mating. Gene flow estimates indicated bidirectionality between all pairs of sampled areas, with magnitudes reflecting clustering and differentiation patterns. A Discriminant Analysis of Principal Components analysis indicated that these genotypic data were best described with four clusters and indicated connectivity across the Saint Lawrence River and Seaway, a potential physical barrier to gene flow. Tests for genetic differentiation indicated restricted gene flow between populations across the Saint Lawrence River and Seaway, and between many sampled areas facing expanding human activity. These results document current genetic variation and connectivity of moose populations in eastern North America, highlight potential challenges to current population connectivity, and identify areas for future research and conservation.

New York, Vermont, New Hampshire, Maine, and Massa

Population genetics of museum specimens indicate decreasing genetic resiliency: The case of two bumble bees of conservation concern

Genetic resiliency is the likelihood that populations retain sufficient genetic diversity to respond to environmental change. It is rarely examined through time in conservation genetic studies due to challenges of acquiring and sequencing historical specimens. Focusing on populations of two sibling bumble bee species of conservation concern with different recent patterns of decline, we used museum specimens collected between 1960 and 2020 and 15 microsatellite markers to assess genetic resiliency (allelic richness, expected heterozygosity, and inbreeding) through time and across geographic space. We find evidence of decreasing allelic richness through time, starting at least 30 years before observed abundance declines in one species and at least 20 years before present in a species with apparently stable abundance. We also found increasing expected heterozygosity through time, indicating increased inbreeding, in the putatively stable species. We demonstrate that genetic measurements taken from specimens collected through time can be used to detect population decline in imperiled species before decreases in abundance are detected. We also demonstrate the importance of interpreting population genetic metrics within the context of historical patterns to assess species' conservation statuses. Finally, we discuss the limitations of currently available population genetic methods, including the influence of isolation by distance and sampling density on measurements of genetic structure, and the influence of demographic characteristics and choice of genetic markers on estimates of genetic diversity and structure. We call for further development of individual-based modeling methods to measure genetic structure, as opposed to commonly applied population-based metrics, to overcome these limitations.

Biological Conservation

Global genetic diversity status and trends: Towards a suite of Essential Biodiversity Variables (EBVs) for genetic composition

Biodiversity underlies ecosystem resilience, ecosystem function, sustainable economies, and human well-being. Understanding how biodiversity sustains ecosystems under anthropogenic stressors and global environmental change will require new ways of deriving and applying biodiversity data. A major challenge is that biodiversity data and knowledge are scattered, biased, collected with numerous methods, and stored in inconsistent ways. The Group on Earth Observations Biodiversity Observation Network (GEO BON) has developed the Essential Biodiversity Variables (EBVs) as fundamental metrics to help aggregate, harmonize, and interpret biodiversity observation data from diverse sources. Mapping and analyzing EBVs can help to evaluate how aspects of biodiversity are distributed geographically and how they change over time. EBVs are also intended to serve as inputs and validation to forecast the status and trends of biodiversity, and to support policy and decision making. Here, we assess the feasibility of implementing Genetic Composition EBVs (Genetic EBVs), which are metrics of within-species genetic variation. We review and bring together numerous areas of the field of genetics and evaluate how each contributes to global and regional genetic biodiversity monitoring with respect to theory, sampling logistics, metadata, archiving, data aggregation, modeling, and technological advances. We propose four Genetic EBVs: ( i ) Genetic Diversity; ( ii ) Genetic Differentiation; ( iii ) Inbreeding; and ( iv ) Effective Population Size ( N e ). We rank Genetic EBVs according to their relevance, sensitivity to change, generalizability, scalability, feasibility and data availability. We outline the workflow for generating genetic data underlying the Genetic EBVs, and review advances and needs in archiving genetic composition data and metadata. We discuss how Genetic EBVs can be operationalized by visualizing EBVs in space and time across species and by forecasting Genetic EBVs beyond current observations using various modeling approaches. Our review then explores challenges of aggregation, standardization, and costs of operationalizing the Genetic EBVs, as well as future directions and opportunities to maximize their uptake globally in research and policy. The collection, annotation, and availability of genetic data has made major advances in the past decade, each of which contributes to the practical and standardized framework for large-scale genetic observation reporting. Rapid advances in DNA sequencing technology present new opportunities, but also challenges for operationalizing Genetic EBVs for biodiversity monitoring regionally and globally. With these advances, genetic composition monitoring is starting to be integrated into global conservation policy, which can help support the foundation of all biodiversity and species' long-term persistence in the face of environmental change. We conclude with a summary of concrete steps for researchers and policy makers for advancing operationalization of Genetic EBVs. The technical and analytical foundations of Genetic EBVs are well developed, and conservation practitioners should anticipate their increasing application as efforts emerge to scale up genetic biodiversity monitoring regionally and globally.

Biological Reviews

High-throughput sequencing reveals distinct regional genetic structure among remaining populations of an endangered salt marsh plant in California

Conservation of rare species requires careful consideration to both preserve locally adapted traits and maintain genetic diversity, as species’ ranges fluctuate in response to a changing climate and habitat loss. Salt marsh systems in California have been highly modified and many salt marsh obligate species have undergone range reductions and habitat loss with concomitant losses of genetic diversity and connectivity. Remaining salt marshes are threatened by rising sea levels, and so these habitats will likely require active restoration and re-establishment efforts. This study aims to provide a reference point for the current status of genetic diversity and range-wide population structure of a federally and state listed endangered plant, Salt Marsh Bird’s Beak (Chloropyron maritimum subsp. maritimum) that can inform future preservation and restoration efforts. We used historical data and current monitoring information to locate and sample all known occurrences throughout the species range in Southern California, and three additional occurrences from Baja California, Mexico. We used flow cytometry and single nucleotide polymorphic markers (SNPs), generated by double-digest restriction-site associated DNA sequencing (ddRAD), to assess relative ploidy, and estimate genetic diversity and population structure across the region. Overall, we found four to five distinct genetic clusters that coincide with geographic regions. Genetic diversity was greatest in the southern part of the range including Baja California and San Diego. These findings can bolster management and restoration efforts by identifying potentially isolated occurrences and areas that are rich sources of allelic diversity, and by providing insight into the amount of genetic differentiation across the species range.

California

Considerations for maximizing the adaptive potential of restored coral populations in the western Atlantic

Active coral restoration typically involves two interventions: crossing gametes to facilitate sexual larval propagation; and fragmenting, growing, and outplanting adult colonies to enhance asexual propagation. From an evolutionary perspective, the goal of these efforts is to establish self‐sustaining, sexually reproducing coral populations that have sufficient genetic and phenotypic variation to adapt to changing environments. Here, we provide concrete guidelines to help restoration practitioners meet this goal for most Caribbean species of interest. To enable the persistence of coral populations exposed to severe selection pressure from many stressors, a mixed provenance strategy is suggested: genetically unique colonies (genets) should be sourced both locally as well as from more distant, environmentally distinct sites. Sourcing three to four genets per reef along environmental gradients should be sufficient to capture a majority of intraspecies genetic diversity. It is best for practitioners to propagate genets with one or more phenotypic traits that are predicted to be valuable in the future, such as low partial mortality, high wound healing rate, high skeletal growth rate, bleaching resilience, infectious disease resilience, and high sexual reproductive output. Some effort should also be reserved for underperforming genets because colonies that grow poorly in nurseries sometimes thrive once returned to the reef and may harbor genetic variants with as yet unrecognized value. Outplants should be clustered in groups of four to six genets to enable successful fertilization upon maturation. Current evidence indicates that translocating genets among distant reefs is unlikely to be problematic from a population genetic perspective but will likely provide substantial adaptive benefits. Similarly, inbreeding depression is not a concern given that current practices only raise first‐generation offspring. Thus, proceeding with the proposed management strategies even in the absence of a detailed population genetic analysis of the focal species at sites targeted for restoration is the best course of action. These basic guidelines should help maximize the adaptive potential of reef‐building corals facing a rapidly changing environment.

Ecological Applications

Population genomic surveys for six rare plant species in San Diego County, California

San Diego County is a hotspot of biodiversity, situated at the intersection of the Baja peninsula, the California floristic province, and the desert southwest. This hotspot is characterized by a high number of rare and endemic species, which persist alongside a major urban epicenter. San Diego County has implemented a strategic management plan that identifies species, based on low numbers of occurrences or high level of threat, for which management practices are recommended. In creating a management plan for rare species, it is important to strike a balance between preserving locally adapted traits and maintaining genetic diversity, as species’ ranges fluctuate in response to a changing climate and habitat fragmentation. This project, in partnership with the San Diego Natural History Museum, aims to provide a reference point for the current status of genetic diversity of rare plant species that will inform future preservation and restoration efforts. We focused on six threatened or endangered plant species: Acanthomintha ilicifolia , Baccharis vanessae , Chloropyron maritimum ssp. maritimum , Deinandra conjugens , Dicranostegia orcuttiana , and Monardella viminea . For each species, botanists from the San Diego Natural History Museum visited all known occurrences in San Diego County and collected leaf tissue for genetic and cytological analysis. We then developed a panel of genetic markers to estimate genetic diversity and population structure. This population genetic survey provided insight into the amount of genetic differentiation across each species’ range, identified isolated occurrences potentially subject to inbreeding or genetic bottlenecks, and identified areas that are rich sources of allelic diversity. Finally, we convened a panel of experts to review results and compatible management options for each species. A summary of the management workshop is included in this report. Overall, we found low genetic differentiation among occurrences across the San Diego region for all species, with the exception of A. ilicifolia . Relative inbreeding was low and consistent across sites, and genetic diversity across sites was variable, with noted exceptions. These findings allow for a wide array of management options that are compatible with panmictic population structure in five of the six surveyed species.

California

Regional differentiation among populations of the Diamondback terrapin ( Malaclemys terrapin )

The Diamondback terrapin (Malaclemys terrapin) is a brackish-water turtle species whose populations have been fragmented due to anthropogenic activity such as development of coastal habitat and entrapment in commercial blue crab (Callinectes sapidus) fishing gear. Genetic analyses can improve conservation efforts for the long-term protection of the species. We used microsatellite DNA analysis to investigate levels of gene flow among and genetic variability within 21 geographically separate collections of the species distributed from Massachusetts to Texas. Quantified levels of genetic variability (allelic diversity, genotypic frequencies, and heterozygosity) revealed three zones of genetic discontinuity, resulting in four discrete populations: Northeast Atlantic, Coastal Mid-Atlantic, Florida and Texas/Louisiana. The average number of alleles and expected heterozygosity for the four genetic clusters were N A = 6.54 and H E = 0.050, respectively. However, the geographic boundaries of the populations did not correspond to accepted terrapin subspecies limits. Our results illuminate not only the need to sample terrapins in additional sites, specifically in the southeast, but also the necessity for allowing uninterrupted gene flow among population groupings to preserve current levels of genetic diversity.

Conservation Genetics

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

Conservation genomics reveals multiple evolutionary units within Bell’s Vireo ( Vireo bellii ).

The Bell’s Vireo ( Vireo bellii ) is a widespread North American species of bird that has declined since the mid-1960s primarily due to habitat modification. Throughout its range, Bell’s Vireo populations are regulated under varying degrees of protection; however, the species has never been characterized genetically. Therefore, the current taxonomy used to guide management decisions may misrepresent the true evolutionary history for the species. We sequenced 86 individuals for ND2 and genotyped 48 individuals for genome-wide SNPs to identify distinct lineages within Bell’s Vireo. Phylogenetic analyses uncovered two distinct clades that are separated in the arid southwestern United States, near the border of the Chihuahuan and Sonoran Deserts. These clades diverged from each other approximately 1.11–2.04 mya. The timing of diversification, geographic location, and niche modeling of the east/west divergence suggest vicariance as a mode of diversification for these two lineages. Analyses of the SNP dataset provided additional resolution and indicated the Least Bell’s Vireo populations are a distinct evolutionary lineage. Our genetic evidence, together with information from morphology and behavior, suggests that the Bell’s Vireo complex involves two species, each containing two separate subspecies. This new information has implications for the federal, state and other listing status of Bell’s Vireo throughout its range.

Arizona, California, Missouri, Nebraska, Nevada, O

Genetic analyses of captive Alala (Corvus hawaiiensis) using AFLP analyses

Population level studies of genetic diversity can provide information about population structure, individual genetic distinctiveness and former population size. They are especially important for rare and threatened species like the Alala, where they can be used to assess extinction risks and evolutionary potential. In an ideal situation multiple methods should be used to detect variation, and these methods should be comparable across studies. In this report, we discuss AFLP (Amplified Fragment Length Polymorphism) as a genetic approach for detecting variation in the Alala , describe our findings, and discuss these in relation to mtDNA and microsatellite data reported elsewhere in this same population. AFLP is a technique for DNA fingerprinting that has wide applications. Because little or no prior knowledge of the particular species is required to carry out this method of analysis, AFLP can be used universally across varied taxonomic groups. Within individuals, estimates of diversity or heterozygosity across genomes may be complex because levels of diversity differ between and among genes. One of the more traditional methods of estimating diversity employs the use of codominant markers such as microsatellites. Codominant markers detect each allele at a locus independently. Hence, one can readily distinguish heterozygotes from homozygotes, directly assess allele frequencies and calculate other population level statistics. Dominant markers (for example, AFLP) are scored as either present or absent (null) so heterozygotes cannot be directly distinguished from homozygotes. However, the presence or absence data can be converted to expected heterozygosity estimates which are comparable to those determined by codominant markers. High allelic diversity and heterozygosity inherent in microsatellites make them excellent tools for studies of wild populations and they have been used extensively. One limitation to the use of microsatellites is that heterozygosity estimates are affected by the mutation rate at microsatellite loci, thus introducing a bias. Also, the number of loci that can be studied is frequently limited to fewer than 10. This theoretically represents a maximum of one marker for each of 10 chromosomes. Dominant markers like AFLP allow a larger fraction of the genome to be screened. Large numbers of loci can be screened by AFLP to resolve very small individual differences that can be used for identification of individuals, estimates of pairwise relatedness and, in some cases, for parentage analyses. Since AFLP is a dominant marker (can not distinguish between +/+ homozygote versus +/- heterozygote), it has limitations for parentage analyses. Only when both parents are homozygous for the absence of alleles (-/-) and offspring show a presence (+/+ or +/-) can the parents be excluded. In this case, microsatellites become preferable as they have the potential to exclude individual parents when the other parent is unknown. Another limitation of AFLP is that the loci are generally less polymorphic (only two alleles/locus) than microsatellite loci (often >10 alleles/locus). While generally fewer than 10 highly polymorphic microsatellite loci are enough to exclude and assign parentage, it might require up to 100 or more AFLP loci. While there are pros and cons to different methodologies, the total number of loci evaluated by AFLP generally offsets the limitations imposed due to the dominant nature of this approach and end results between methods are generally comparable. Overall objectives of this study were to evaluate the level of genetic diversity in the captive population of Alala, to compare genetic data with currently available pedigree information, and to determine the extent of relatedness of mating pairs and among founding individuals.

Open-File Report

Next‐generation conservation genetics and biodiversity monitoring

This special issue of Evolutionary Applications consists of 10 publications investigating the use of next‐generation tools and techniques in population genetic analyses and biodiversity assessment. The special issue stems from a 2016 Next Generation Genetic Monitoring Workshop, hosted by the National Institute for Mathematical and Biological Synthesis (NIMBioS) in Tennessee, USA. The improved accessibility of next‐generation sequencing platforms has allowed molecular ecologists to rapidly produce large amounts of data. However, with the increased availability of new genomic markers and mathematical techniques, care is needed in selecting appropriate study designs, interpreting results in light of conservation concerns, and determining appropriate management actions. This special issue identifies key attributes of successful genetic data analyses in biodiversity evaluation and suggests ways to improve analyses and their application in current population and conservation genetics research.

Evolutionary Applications

Sampling across 20 years (1996–2017) reveals loss of diversity and genetic connectivity in the Coachella Valley fringe-toed lizard ( Uma inornata )

The Coachella Valley fringe-toed lizard ( Uma inornata ) is a federally threatened, aeolian sand dune obligate, endemic to the Coachella Valley, California. Historically, U. inornata is thought to have formed a large interconnected metapopulation across the valley, with local dune habitat and population size fluctuations linked to stochastic droughts and flooding. Since the 1950s, aeolian habitat in Coachella Valley has declined by 91–95 percent. What remains is highly fragmented by highways and development in the urban communities of the Coachella Valley, raising concerns that fringe-toed lizard movement and gene flow among remaining habitat fragments is limited or nonexistent. We examined population genetic structure across three sample periods (1996, 2008, and 2017). Over that time, this species has shifted from a panmictic condition (1996) with little or no genetic structure between sites to the current (2017) condition where there are now genetically distinct populations. Two severe droughts (2000–04 and 2012–16) may have accelerated this shift through drought-related population declines and subsequent genetic bottlenecks. Using a combination of microsatellite loci and single nucleotide polymorphisms, we found patterns of decreasing genetic connectivity and diversity over time. These patterns are consistent with reduced fringe-toed lizard movement and gene flow among isolated sand dune systems. Low effective population sizes were recovered in some sites, suggesting genetic drift in smaller and fluctuating populations is likely responsible for loss of genetic diversity. A U.S. Fish and Wildlife Service recovery objective for this species is to maintain genetic diversity; however, evidence of fragmentation suggests that genetic cohesiveness has been altered and that the diversity maintained in individual fragments is lower than in the total metapopulation. Management actions that increase genetic diversity could be implemented, including translocation. We modeled increasing gene flow between 1–10 percent, which showed that allelic richness could increase rapidly if translocated individuals can survive and reproduce. Establishing translocation protocols could help to avoid the high mortality that has occurred with other reptile translocations. Successful translocations could be a useful strategy to replenish lost genetic diversity after bottlenecks and could mitigate the loss of natural gene flow among populations.

California

Genetic conservation and paddlefish propagation

The conservation of genetic diversity of our natural resources is overwhelmingly one of the central foci of 21st century management practices. Three recommendations related to the conservation of paddlefish Polyodon spathula genetic diversity are to (1) identify genetic diversity at both nuclear and mitochondrial DNA loci using a suggested list of 20 sampling locations, (2) use genetic diversity estimates to develop genetic management units, and (3) identify broodstock sources to minimize effects of supplemental stocking on the genetic integrity of native paddlefish populations. We review previous genetic work on paddlefish and described key principles and concepts associated with maintaining genetic diversity within and among paddlefish populations and also present a genetic case study of current paddlefish propagation at the U.S. Fish and Wildlife Service Gavins Point National Fish Hatchery. This study confirmed that three potential sources of broodfish were genetically indistinguishable at the loci examined, allowing the management agencies cooperating on this program flexibility in sampling gametes. This study also showed significant bias in the hatchery occurred in terms of male reproductive contribution, which resulted in a shift in the genetic diversity of progeny compared to the broodfish. This shift was shown to result from differential male contributions, partially attributed to the mode of egg fertilization. Genetic insights enable implementation of a paddlefish propagation program within an adaptive management strategy that conserves inherent genetic diversity while achieving demographic goals.

American Fisheries Society Symposium

Characterization of the genetic structure of four sucker species in the Klamath River. Final Report

Four species of suckers (family Catostomidae) inhabit the Klamath River Basin of Oregon and California: Lost River suckers (LRS; Deltistes luxatus), shortnose suckers (SNS; Chasmistes brevirostris), Klamath largescale suckers (KLS; Catostomus snyderi), and Klamath smallscale suckers (KSS; Catostomus rimiculus). All but Klamath smallscale suckers are endemic and restricted to the Klamath River Basin where they occur sympatrically in large lakes and reservoirs, including the Lost River and Klamath Lake subbasins (Figure 1; USFWS 2012). Population declines, primarily due to loss or degradation of spawning, rearing, and adult habitat, have resulted in Lost River and shortnose suckers being listed as endangered throughout their entire range under the U.S. Endangered Species Act (USFWS 1988). Continued population declines coupled with failed adult recruitment prompted the USFWS to initiate an assisted rearing program in 2015 as a part of their recovery strategy (Childress et al. 2019). The program was designed to maintain as much genetic diversity as possible while improving recruitment by averting high early life stage mortality (Day et al. 2017). However, while assisted rearing efforts are targeted towards endangered LRS and SNS, species differentiation of larval and juvenile suckers is problematic in the Klamath River Basin. This, in turn, complicates the management of these species as well as the population modeling used to evaluate recovery efforts. Maintaining as much as possible of the genetic resources, or “evolutionary legacy” of a species is a goal common to conservation and endangered species recovery strategies. Inappropriate assumptions regarding species’ evolutionary lineages, and genetic characteristics may lead to the mismanagement of an endangered species through a failure to recognize and appropriately manage species boundaries and genetic population structure. Despite a considerable amount of research, the partitioning of genetic diversity within and among the four species of suckers in the Klamath River Basin remains unclear. Previously developed genetic markers are effective at differentiating some species, but fail to effectively differentiate all four species of suckers in the basin (Tranah et al. 2001; Wagman 2003; Tranah and May 2006; Hoy and Ostberg 2015; Dowling et al. 2016). Peer-reviewed publications describing the morphological characteristics of (Markle et al. 2005) and genetic relationships among (Dowling et al. 2016; Tranah and May 2006) Klamath River Basin suckers have not resolved uncertainties regarding the systematic relationships among the four currently recognized taxa. Specifically, genetic and morphological data generally support LRS and KSS as being distinct entities, but genetic evidence does not support a distinction between KLS and SNS. All three publications above refer to unpublished information regarding ecological differences between KLS and SNS as evidence to support the existence of two entities. However, the authors also acknowledge that overlap in morphological characters (Markle et al. 2005) and a lack of genetic differentiation (Dowling et al. 2016; Tranah and May 2006) between KLS and SNS raises the question of their specific identity. This is particularly problematic in the Lost River subbasin, where overlap in morphological characters between KLS and SNS is greatest. In our opinion, the basis of the strong genetic similarity between KLS and SNS has not been resolved. Morphological characters mostly support the existence of two distinct species, while genetic characters do not (i.e., genetic divergence between KLS and SNS is less than divergence among populations of each species; (Smith et al. 2015). Some have suggested that introgressive hybridization may have resulted in a lack of genetic differentiation between KLS and SNS and a breakdown of monophyletic species (Dowling et al. 2016; Tranah and May 2006), and that this hybridization may be an important process in

California, Oregon

Atlantic salmon (Salmo salar) culture manual

The primary objective of the Atlantic Salmon Research Program established at the U.S. Geological Survey Tunison Laboratory of Aquatic Science as mandated by the Great Lakes Restoration Initiative is to restore Atlantic salmon (Linnaeus, 1758; Salmo salar ) into Lake Ontario. This objective focuses on evaluating the survival of stocked Atlantic salmon in current Lake Ontario conditions to create a genetic strain that is robust to overcome current physiological and physical barriers. To complete this goal, new and innovative hatchery techniques based on past successful methods and protocols to grow Atlantic salmon to various life stages are described in this standard operating manual.

Techniques and Methods

Disentangling genetic diversity of Myotis septentrionalis: population structure, demographic history, and effective population size

Myotis septentrionalis (Northern Long-eared Bat) has recently suffered a >90% decline in population size in North America due to white-nose syndrome (WNS). We assessed genetic diversity, population structure, current effective population size, and demographic history of M. septentrionalis distributed across the United States to determine baseline levels pre-WNS. We analyzed RADseq data from 81 individuals from Kentucky, Louisiana, Michigan, Minnesota, North Carolina, Oklahoma, and Wisconsin. Additionally, we examined population genetic structure using discriminant analysis of principal components, fastStructure, and STRUCTURE. We then estimated effective population size and demographic history using fastsimcoal2. Similar levels of genetic diversity were found across all samples. We found no population genetic structure in the varied analyses from these contemporary samples. The best model for demographic history estimated a rapid population expansion followed by a slower expansion approximately 340,000 years ago. The vagility of M. septentrionalis , along with male dispersal and random mating, may provide a buffer against serious bottleneck effects stemming from rapid population declines due to WNS. This research provides a baseline for tracking and monitoring the influence of WNS on genetic diversity such as potential reduced diversity or increased population structuring in the future.

Kentucky, Louisiana, Michigan, Minnesota, North Ca