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Application of genetics and genomics to wildlife epidemiology

Wildlife diseases can have significant impacts on wildlife conservation and management. Many of the pathogens that affect wildlife also have important implications for domestic animal and human health. However, management interventions to prevent or control wildlife disease are hampered by uncertainties about the complex interactions between pathogens and free-ranging wildlife. We often lack crucial knowledge about host ecology, pathogen characteristics, and host–pathogen dynamics. The purpose of this review is to familiarize wildlife biologists and managers with the application of genetic and genomic methodologies for investigating pathogen and host biology to better understand and manage wildlife diseases. The genesis of this review was a symposium at the 2013 annual Wildlife Society Conference. We reviewed the scientific literature and used our personal experiences to identify studies that illustrate the application of genetic and genomic methods to advance our understanding of wildlife epidemiology, focusing on recent research, new techniques, and innovative approaches. Using examples from a variety of pathogen types and a broad array of vertebrate taxa, we describe how genetics and genomics can provide tools to detect and characterize pathogens, uncover routes of disease transmission and spread, shed light on the ways that disease susceptibility is influenced by both host and pathogen attributes, and elucidate the impacts of disease on wildlife populations. Genetic and increasingly genomic methodologies will continue to contribute important insights into pathogen and host biology that will aid efforts to assess and mitigate the impacts of wildlife diseases on global health and conservation of biodiversity.

Journal of Wildlife Management

Extensive dispersal of Roanoke logperch ( Percina rex ) inferred from genetic marker data

The dispersal ecology of most stream fishes is poorly characterised, complicating conservation efforts for these species. We used microsatellite DNA marker data to characterise dispersal patterns and effective population size ( N e ) for a population of Roanoke logperch Percina rex , an endangered darter (Percidae). Juveniles and candidate parents were sampled for 2 years at sites throughout the Roanoke River watershed. Dispersal was inferred via genetic assignment tests (ATs), pedigree reconstruction (PR) and estimation of lifetime dispersal distance under a genetic isolation-by-distance model. Estimates of N e varied from 105 to 1218 individuals, depending on the estimation method. Based on PR, polygamy was frequent in parents of both sexes, with individuals spawning with an average of 2.4 mates. The sample contained 61 half-sibling pairs, but only one parent–offspring pair and no full-sib pairs, which limited our ability to discriminate natal dispersal of juveniles from breeding dispersal of their parents between spawning events. Nonetheless, all methods indicated extensive dispersal. The AT indicated unrestricted dispersal among sites ≤15 km apart, while siblings inferred by the PR were captured an average of 14 km and up to 55 km apart. Model-based estimates of median lifetime dispersal distance (6–24 km, depending on assumptions) bracketed AT and PR estimates, indicating that widely dispersed individuals do, on average, contribute to gene flow. Extensive dispersal of P. rex suggests that darters and other small benthic stream fishes may be unexpectedly mobile. Monitoring and management activities for such populations should encompass entire watersheds to fully capture population dynamics.

Ecology of Freshwater Fish

Genomic single-nucleotide polymorphisms confirm that Gunnison and Greater sage-grouse are genetically well differentiated and that the Bi-State population is distinct

Sage-grouse are iconic, declining inhabitants of sagebrush habitats in western North America, and their management depends on an understanding of genetic variation across the landscape. Two distinct species of sage-grouse have been recognized, Greater ( Centrocercus urophasianus ) and Gunnison sage-grouse ( C. minimus ), based on morphology, behavior, and variation at neutral genetic markers. A parapatric group of Greater Sage-Grouse along the border of California and Nevada ("Bi-State") is also genetically distinct at the same neutral genetic markers, yet not different in behavior or morphology. Because delineating taxonomic boundaries and defining conservation units is often difficult in recently diverged taxa and can be further complicated by highly skewed mating systems, we took advantage of new genomic methods that improve our ability to characterize genetic variation at a much finer resolution. We identified thousands of single-nucleotide polymorphisms (SNPs) among Gunnison, Greater, and Bi-State sage-grouse and used them to comprehensively examine levels of genetic diversity and differentiation among these groups. The pairwise multilocus fixation index ( F ST ) was high (0.49) between Gunnison and Greater sage-grouse, and both principal coordinates analysis and model-based clustering grouped samples unequivocally by species. Standing genetic variation was lower within the Gunnison Sage-Grouse. The Bi-State population was also significantly differentiated from Greater Sage-Grouse, albeit more weakly ( F ST = 0.09), and genetic clustering results were consistent with reduced gene flow with Greater Sage-Grouse. No comparable genetic divisions were found within the Greater Sage-Grouse sample, which spanned the southern half of the range. Thus, we provide much stronger genetic evidence supporting the recognition of Gunnison Sage-Grouse as a distinct species with low genetic diversity. Further, our work confirms that the Bi-State population is differentiated from other Greater Sage-Grouse. The level of differentiation is much less than the divergence between Greater and Gunnison sage-grouse, supporting the idea that the Bi-State represents a unique population within the Greater Sage-Grouse. New genomic methods like the restriction-site-associated DNA (RAD-tag) method used here illustrate how increasing the number of markers and coverage of the genome can better characterize patterns of genetic variation, particularly among recently diverged taxa, providing vital information for conservation and management.

The Condor

Ecology of Greater Sage-Grouse in the Bi-State Planning Area Final Report, September 2007

Conservation efforts for greater sage-grouse (Centrocercus urophasianus), hereafter sage-grouse, are underway across the range of this species. Over 70 local working groups have been established and are implementing on-the-ground sage-grouse oriented conservation projects. Early on in this process, the California Department of Fish and Game (CDFG) recognized the need to join in these efforts and received funding from the U.S. Fish and Wildlife Service (USFWS) under the Candidate Species Conservation Program to help develop a species conservation plan for sage-grouse in the Mono County area. This conservation plan covers portions of Alpine, Mono, and Inyo counties in California and Douglas, Esmeralda, Lyon, and Mineral counties in Nevada. A concurrent effort underway through the Nevada Governor's Sage-grouse Conservation Team established Local Area Working Groups across Nevada and eastern California. The Mono County populations of sage-grouse were encompassed by the Bi-State Local Planning Area, which was comprised of six population management units (PMUs). The state agencies from California (CDFG) and Nevada (Nevada Department of Wildlife; NDOW) responsible for the management of sage-grouse agreed to utilize the process that had begun with the Nevada Governor's Team in order to develop local plans for conservation planning and implementation. Resources from the USFWS were applied to several objectives in support of the development of the Bi-State Local Area Sage-grouse Conservation Plan through a grant to the U.S. Geological Survey (USGS). Objectives included: (1) participate in the development of the Bi-State Conservation Plan, (2) compile and synthesize existing sage-grouse data, (3) document seasonal movements of sage-grouse, (4) identify habitats critical to sage-grouse, (5) determine survival rates and identify causal factors of mortality, (6) determine nest success and brood success of sage-grouse, and (7) identify sage-grouse lek sites. Progress reports completed in 2004 and 2005 addressed each of the specific objectives and this final report focuses on the biological information gathered in support of local conservation efforts. Participation in the development of the Bi-State Local Area Conservation Plan was accomplished on multiple scales. Beginning in the fall of 2002, USGS personnel began participating in meetings of local stakeholders involved in the development of a sage-grouse conservation plan for the Bi-State planning area. This included attendance at numerous local PMU group meetings and field trips as well as participating on the technical advisory committee (TAC) for the Bi-State group. Whenever appropriate, ongoing results and findings regarding sage-grouse ecology in the local area were incorporated into these working group meetings. In addition, the USGS partnered with CDFG to help reorganize one of the local PMU groups (South Mono) and edited that portion of the Bi-State plan. The USGS also worked closely with CDFG to draft a description of the state of knowledge for sage-grouse genetic information for inclusion in the Bi-State Conservation Plan. The first edition of the Bi-State Conservation Plan for Greater Sage-Grouse was completed in June 2004 (Bi-State Sage-grouse Conservation Team 2004). This report is organized primarily by PMU to facilitate the incorporation of these research findings into the individual PMU plans that compose the Bi-State plan. Information presented in this report was derived from over 7,000 radio-telemetry locations obtained on 145 individual sage-grouse during a three year period (2003-2005). In addition, we collected detailed vegetation measurements at over 590 habitat sampling plots within the study area including canopy cover, shrubs, forbs, and grasses diversity. Vegetation data collection focused on sage-grouse nests, and brood-use areas. Additionally we collected data at random sites to examine sage-grouse habitat relationships within the study area. The majori

Open-File Report

Hybridization and population genetics of Alligator Gar in Lake Texoma

The Alligator Gar Atractosteus spatula (AG) is a long-lived fish of growing management and conservation interest. Situated on the border of Texas and Oklahoma, Lake Texoma supports one of the last robust AG populations in Oklahoma; however, a genetic evaluation of this population is lacking. We genotyped AG individuals with 17 microsatellite loci, 7 of which also cross-amplified in three sympatric Lepisosteus species: the Longnose Gar L. osseus (LN), Shortnose Gar L. platostomus (SN), and Spotted Gar L. oculatus (SP). Bayesian assignment analyses conducted in STRUCTURE and NewHybrids confirmed that a field-identified hybrid was an F 1 AG × LN and identified five other individuals that were suspected backcrosses (three LN × SN; two SN × SP). Alligator Gar had the lowest observed heterozygosity (0.179) and the lowest allelic richness (1.682) among the nonhybrid individuals of the four gar species examined. We also examined the potential for population structure and differences in pairwise relatedness ( r ) between two areas where AG are commonly encountered within Lake Texoma: the Red River and Washita River arms. No population structure was detected using noninformative or location priors in STRUCTURE, and estimates of r produced by the TrioML estimator in COANCESTRY were not significantly different between arms (overall mean r = 0.199). Point estimates of effective population size ranging from 16.3 to 29.2 suggested that the AG population may be vulnerable to the effects of inbreeding depression and random genetic drift. Results provide a genetic status assessment of AG in Lake Texoma and a baseline for future management and conservation decisions within Lake Texoma and surrounding regions.

Oklahoma, Texas

Range-wide genetic analysis of an endangered bumble bee (Bombus affinis, Hymenoptera: Apidae) reveals population structure, isolation by distance, and low colony abundance

Declines in bumble bee species range and abundances are documented across multiple continents and have prompted the need for research to aid species recovery and conservation. The rusty patched bumble bee ( Bombus affinis ) is the first federally listed bumble bee species in North America. We conducted a range-wide population genetics study of B. affinis from across all extant conservation units to inform conservation efforts. To understand the species’ vulnerability and help establish recovery targets, we examined population structure, patterns of genetic diversity, and population differentiation. Additionally, we conducted a site-level analysis of colony abundance to inform prioritizing areas for conservation, translocation, and other recovery actions. We find substantial evidence of population structuring along an east-to-west gradient. Putative populations show evidence of isolation by distance, high inbreeding coefficients, and a range-wide male diploidy rate of ~15%. Our results suggest the Appalachians represent a genetically distinct cluster with high levels of private alleles and substantial differentiation from the rest of the extant range. Site-level analyses suggest low colony abundance estimates for B. affinis compared to similar datasets of stable, co-occurring species. These results lend genetic support to trends from observational studies, suggesting that B. affinis has undergone a recent decline and exhibit substantial spatial structure. The low colony abundances observed here suggest caution in overinterpreting the stability of populations even where B. affinis is reliably detected interannually. These results help delineate informed management units, provide context for the potential risks of translocation programs, and help set clear recovery targets for this and other threatened bumble bee species.

Iowa, Illinois, Indiana, Kentucky, Michigan, Minne

Low genetic variation and evidence of limited dispersal in the regionally important Belize manatee

The Antillean subspecies of the West Indian manatee Trichechus manatus is found throughout Central and South America and the Caribbean. Because of severe hunting pressure during the 17th through 19th centuries, only small populations of the once widespread aquatic mammal remain. Fortunately, protections in Belize reduced hunting in the 1930s and allowed the country's manatee population to become the largest breeding population in the Wider Caribbean. However, increasing and emerging anthropogenic threats such as coastal development, pollution, watercraft collision and net entanglement represent challenges to this ecologically important population. To inform conservation and management decisions, a comprehensive molecular investigation of the genetic diversity, relatedness and population structure of the Belize manatee population was conducted using mitochondrial and microsatellite DNA. Compared with other mammal populations, a low degree of genetic diversity was detected ( H E =0.455; N A =3.4), corresponding to the small population size and long-term exploitation. Manatees from the Belize City Cayes and Southern Lagoon system were genetically different, with microsatellite and mitochondrial F ST values of 0.029 and 0.078, respectively ( P ≤0.05). This, along with the distinct habitats and threats, indicates that separate protection of these two groups would best preserve the region's diversity. The Belize population and Florida subspecies appear to be unrelated with microsatellite and mitochondrial F ST values of 0.141 and 0.63, respectively ( P ≤0.001), supporting the subspecies designations and suggesting low vagility throughout the northern Caribbean habitat. Further monitoring and protection may allow an increase in the Belize manatee genetic diversity and population size. A large and expanding Belize population could potentially assist in the recovery of other threatened or functionally extinct Central American Antillean manatee populations.

Animal Conservation

Estimating bighorn sheep (Ovis canadensis) abundance using noninvasive sampling at a mineral lick within a National Park Wilderness Area

Conservation of species requires accurate population estimates. We used genetic markers from feces to determine bighorn sheep abundance for a herd that was hypothesized to be declining and in need of population status monitoring. We sampled from a small but accessible portion of the population's range where animals naturally congregate at a natural mineral lick to test whether we could accurately estimate population size by sampling from an area where animals concentrate. We used mark-recapture analysis to derive population estimates, and compared estimates from this smaller spatial sampling to estimates from sampling of the entire bighorn sheep range. We found that estimates were somewhat comparable; in 2009, the mineral lick sample and entire range sample differed by 20 individuals, and in 2010 they differed by only one individual. However, we captured 13 individuals in the entire range sample that were not captured at the mineral lick, and thus violated a model assumption that all individuals had an equal opportunity of being captured. This eliminated the possibility of inferring a total population estimate from just animals visiting the mineral lick, but because estimates were relatively similar, monitoring at the mineral lick can provide a useful index for management and conservation. We compared our results to a radio-collar study conducted in 2003–2004 and confirmed that the population remained stable since 2004. Our population estimates were 78 (CI 62–114) in 2009 and 95 (CI 77–131) in 2010. Between 7 and 11 sampling dates were needed to achieve a CV of 20% for population estimates, assuming a capture probability between 0.09 and 0.13. We relied on citizen science volunteers to maximize data collection and reduce costs; 71% of all fecal samples were collected by volunteers, compared to 29% collected by paid staff. We conclude that our technique provides a useful monitoring tool for managers. The technique could be tested and applied in similar populations where animals congregate with high fidelity at a mineral lick or other area.

Colorado

Are captive tortoises a reservoir for conservation? An assessment of genealogical affiliation of captive Gopherus agassizii to local, wild populations

The conservation of tortoises poses a unique situation because several threatened species are commonly kept as pets within their native ranges. Thus, there is potential for captive populations to be a reservoir for repatriation efforts. We assess the utility of captive populations of the threatened Agassiz’s desert tortoise ( Gopherus agassizii ) for recovery efforts based on genetic affinity to local areas. We collected samples from 130 captive desert tortoises from three desert communities: two in California (Ridgecrest and Joshua Tree) and the Desert Tortoise Conservation Center (Las Vegas) in Nevada. We tested all samples for 25 short tandem repeats and sequenced 1,109 bp of the mitochondrial genome. We compared captive genotypes to a database of 1,258 Gopherus samples, including 657 wild caught G. agassizii spanning the full range of the species. We conducted population assignment tests to determine the genetic origins of the captive individuals. For our total sample set, only 44 % of captive individuals were assigned to local populations based on genetic units derived from the reference database. One individual from Joshua Tree, California, was identified as being a Morafka’s desert tortoise, G. morafkai , a cryptic species which is not native to the Mojave Desert. Our data suggest that captive desert tortoises kept within the native range of G. agassizii cannot be presumed to have a genealogical affiliation to wild tortoises in their geographic proximity. Precautions should be taken before considering the release of captive tortoises into the wild as a management tool for recovery.

Conservation Genetics

Determining the distribution, status, and linkages of Agassiz's desert tortoise populations in the uplands surrounding the Coachella Valley

In support of the goals of the Coachella Valley Conservation Commission and the Bureau of Land Management, we performed surveys to determine the status, distribution, demographics, and possible genetic linkages of Agassiz’s desert tortoise (Gopherus agassizii) populations within the Coachella Valley Multiple Species Habitat Conservation Plan (CVMSHCP) area during a multi-decadal megadrought. Federal, state, and numerous parcels of privately owned conservation lands in the uplands around the periphery of the Coachella Valley were surveyed. Desert tortoise distribution data collected will be used to develop a species habitat model. These data will be overlaid with location data of flammable invasive plant species to inform fuels management and invasive plant control efforts in sensitive species habitat, as well as continue to identify high quality habitat and important desert tortoise linkage areas around the Coachella Valley. We also surveyed two established G. agassizii study plots – one at the Mesa Wind Energy Facility near Palm Springs, California and one at the Boyd Deep Canyon Desert Research Center. At those locations, we used resurveys and radio telemetry to assess habitat use, demography, and population status. We also used camera trapping at Deep Canyon to observe tortoise activities and behaviors in an unusual low elevation tortoise habitat.

California

Where east meets west: Phylogeography of the high Arctic North American brant goose

Genetic variation in Arctic species is often influenced by vicariance during the Pleistocene, as ice sheets fragmented the landscape and displaced populations to low- and high-latitude refugia. The formation of secondary contact or suture zones during periods of ice sheet retraction has important consequences on genetic diversity by facilitating genetic connectivity between formerly isolated populations. Brant geese ( Branta bernicla ) are a maritime migratory waterfowl (Anseriformes) species that almost exclusively uses coastal habitats. Within North America, brant geese are characterized by two phenotypically distinct subspecies that utilize disjunct breeding and wintering areas in the northern Pacific and Atlantic. In the Western High Arctic of Canada, brant geese consist of individuals with an intermediate phenotype that are rarely observed nesting outside this region. We examined the genetic structure of brant geese populations from each subspecies and areas consisting of intermediate phenotypes using mitochondrial DNA (mtDNA) control region sequence data and microsatellite loci. We found a strong east–west partition in both marker types consistent with refugial populations. Within subspecies, structure was also observed at mtDNA while microsatellite data suggested the presence of only two distinct genetic clusters. The Western High Arctic (WHA) appears to be a secondary contact zone for both Atlantic and Pacific lineages as mtDNA and nuclear genotypes were assigned to both subspecies, and admixed individuals were observed in this region. The mtDNA sequence data outside WHA suggests no or very restricted intermixing between Atlantic and Pacific wintering populations which is consistent with published banding and telemetry data. Our study indicates that, although brant geese in the WHA are not a genetically distinct lineage, this region may act as a reservoir of genetic diversity and may be an area of high conservation value given the potential of low reproductive output in this species.

Ecology and Evolution

Glacial vicariance and secondary contact shape demographic histories in a freshwater mussel species complex

Characterizing the mechanisms influencing the distribution of genetic variation in aquatic species can be difficult due to the dynamic nature of hydrological landscapes. In North America’s Central Highlands, a complex history of glacial dynamics, long-term isolation, and secondary contact have shaped genetic variation in aquatic species. Although the effects of glacial history have been demonstrated in many taxa, responses are often lineage- or species-specific and driven by organismal ecology. In this study, we reconstruct the evolutionary history of a freshwater mussel species complex using a suite of mitochondrial and nuclear loci to resolve taxonomic and demographic uncertainties. Our findings do not support Pleurobema rubrum as a valid species, which is proposed for listing as threatened under the U.S. Endangered Species Act. We synonymize P. rubrum under Pleurobema sintoxia —a common and widespread species found throughout the Mississippi River Basin. Further investigation of patterns of genetic variation in P. sintoxia identified a complex demographic history, including ancestral vicariance and secondary contact, within the Eastern Highlands. We hypothesize these patterns were shaped by ancestral vicariance driven by the formation of Lake Green and subsequent secondary contact after the last glacial maximum. Our inference aligns with demographic histories observed in other aquatic taxa in the region and mirrors patterns of genetic variation of a freshwater fish species ( Erimystax dissimilis ) confirmed to serve as a parasitic larval host for P. sintoxia . Our findings directly link species ecology to observed patterns of genetic variation and may have significant implications for future conservation and recovery actions of freshwater mussels.

Mississippi River basin

Spatial segregation between phenotypes of the diablotin black-capped petrel Pterodroma hasitata during the non-breeding period

Despite growing support for ecosystem-based approaches, conservation is mostly implemented at the species level. However, genetic differentiation exists within this taxonomic level, putting genetically distinct populations at risk of local extinction. In the diablotin black-capped petrel Pterodroma hasitata , an endangered gadfly petrel endemic to the Caribbean, 2 phenotypes have been described: a smaller dark form and a heavier light form, which are genetically distinct. To assess possible differences in the marine distributions of phenotypes, in May 2019, we captured 5 adult black-capped petrels of each phenotype at sea in the western North Atlantic and equipped them with satellite transmitters. We used generalized linear mixed models to test the importance of phenotype on geographic distribution. Using kernel density estimations, we located use areas, quantified spatial overlap between forms, and assessed form-specific exposure to marine threats. Petrels were tracked for 11 to 255 d ( mean ± SD: 102.1 ± 74.2 d). During the non-breeding period, all individuals ranged from 28.4 to 43.0° latitude. Phenotypes had significantly distinct non-breeding distributions, independent of time of year. The dark form used waters of the Carolinian marine ecoregion, and the light form used pelagic waters of the Virginian ecoregion, to the north. The dark form was more exposed to marine threats than the light form, in particular to mercury, microplastics, and marine traffic. The light form overlapped with proposed wind energy areas off the central US coast. These differences in exposure suggest possible differences in vulnerability, which can have repercussions on the viability of this imperiled species.

North Carolina

Genetic structure and historic demography of endangered unarmoured threespine stickleback at southern latitudes signals a potential new management approach

Habitat loss, flood control infrastructure, and drought have left most of southern California and northern Baja California's native freshwater fish near extinction, including the endangered unarmoured threespine stickleback ( Gasterosteus aculeatus williamsoni ). This subspecies, an unusual morph lacking the typical lateral bony plates of the G. aculeatus complex, occurs at arid southern latitudes in the eastern Pacific Ocean and survives in only three inland locations. Managers have lacked molecular data to answer basic questions about the ancestry and genetic distinctiveness of unarmoured populations. These data could be used to prioritize conservation efforts. We sampled G. aculeatus from 36 localities and used microsatellites and whole genome data to place unarmoured populations within the broader evolutionary context of G. aculeatus across southern California/northern Baja California. We identified three genetic groups with none consisting solely of unarmoured populations. Unlike G. aculeatus at northern latitudes, where Pleistocene glaciation has produced similar historical demographic profiles across populations, we found markedly different demographics depending on sampling location, with inland unarmoured populations showing steeper population declines and lower heterozygosity compared to low armoured populations in coastal lagoons. One exception involved the only high elevation population in the region, where the demography and alleles of unarmoured fish were similar to low armoured populations near the coast, exposing one of several cases of artificial translocation. Our results suggest that the current “management-by-phenotype” approach, based on lateral plates, is incidentally protecting the most imperilled populations; however, redirecting efforts toward evolutionary units, regardless of phenotype, may more effectively preserve adaptive potential.

Baja California, California

Evaluating the central–marginal hypothesis: Introgression and genetic variation at the trailing edge of Quercus bicolor

The central–marginal hypothesis (CMH) predicts reduced genetic diversity and increased differentiation in range-edge populations due to ecological marginality and limited gene flow. Deviations from this pattern, however, can result from historical demographic processes, variation in reproductive strategies or interspecific hybridization. The genus Quercus , known for hybridization and long-distance pollination, offers an excellent model to examine the spatial patterns of genetic diversity, structure and introgression across species distributions. Here, we investigate these dynamics in Quercus bicolor Willd., a widespread eastern North American oak. Using RADseq, we genotyped 142 individuals from 12 sites at the fragmented trailing range edge and nine sites from the range core. To detect introgression, we incorporated reference data from six sympatric white oak species. We reveal extensive introgression, particularly from Q. lyrata Walt., in nearly all southern edge populations, but none in core populations despite sympatry with closely related congeners. Southern populations also showed increased genetic structure and differentiation, but not reduced diversity or increased inbreeding, even when only examining non-admixed individuals. Regression analyses reveal relationships between introgressed ancestry and heterozygosity, inbreeding and differentiation, indicating that introgression may buffer range-edge populations against genetic erosion by introducing novel alleles. Hindcast, current and forecast ecological niche models demonstrate temporally changing degrees of overlap between the geographic range of Q. lyrata and Q. bicolor and suggest higher hybridization potential in the future. These findings offer mixed support for the CMH while underscoring the evolutionary relevance of introgression in shaping genetic landscapes at range margins with significant implications for conservation.

Molecular Ecology

Where has turtle ecology been, and where is it going?

Over 9000 articles have been published on turtles and tortoises (excluding sea turtles) since 1950 according to the Web of Science, including over 8000 contained in a personal bibliography that we analyze in this paper. Research had a slow start from 1900 to 1950, with mostly anecdotal additions to our knowledge until the contributions of F. Cagle and A. Carr took turtle research to new levels as the cofathers of turtle ecology in the middle of the last century. Books written in 1939, 1952, and 1972 that compiled existing literature on turtles in the United States and Canada set the stage for growing interest in turtles. The first global compilation of turtle species did not become available until 1961. Publication frequency increased in the 1960s and especially the 1970s as interest in turtles grew, and a wave of turtle biologists emerged from doctoral degree programs. We briefly review the contributions of scientists who published extensively on turtle ecology in those and later decades up to the present. We also review advances in our knowledge of various topics, including the global distribution of turtle research efforts; changes in our perceptions of turtle species diversity over time; turtle community ecology; sex ratios, sex-determination, and climate change; overwintering behavior; sexual size dimorphism and sexual dichromatism; analyses of population genetics; turtles and vocalization; and the emergence of turtle conservation biology efforts. We conclude with a discussion of future opportunities and challenges for working with turtles.

Herpetologica

The distribution of nuclear genetic variation and historical demography of sea otters

The amount and distribution of population genetic variation is crucial information for the design of effective conservation strategies for endangered species and can also be used to provide inference about demographic processes and patterns of migration. Here, we describe variation at a large number of nuclear genes in sea otters Enhydra lutris ssp. We surveyed 14 variable microsatellite loci and two genes of the major histocompatibility complex (MHC) in up to 350 California sea otters Enhydra lutris nereis , which represents ∼10% of the subspecies' population, and 46 otters from two Alaskan sites. We utilized methods for detecting past reductions in effective population size to examine the effects of near extinction from the fur trade. Summary statistic tests largely failed to find a signal of a recent population size reduction (within the past 200 years), but a Bayesian method found a signal of a strong reduction over a longer time scale (up to 500 years ago). These results indicate that the reduction in size began long enough ago that much genetic variation was lost before the 19th century fur trade. A comparison of geographic distance and pairwise relatedness for individual otters found no evidence of kin-based spatial clustering for either gender. This indicates that there is no population structure, due to extended family groups, within the California population. A survey of population genetic variation found that two of the MHC genes, DQB and DRB, had two alleles present and one of the genes, DRA, was monomorphic in otters. This contrasts with other mammals, where they are often the most variable coding genes known. Genetic variation in the sea otter is among the lowest observed for a mammal and raises concerns about the long-term viability of the species, particularly in the face of future environmental changes.

Animal Conservation

Genetic diversity and structure from Antillean manatee (Trichechus manatus manatus) in the southern Gulf of Mexico: Comparison between connected and isolated populations

Antillean manatees ( Trichechus manatus manatus ), a subspecies of the West Indian manatee, is listed as endangered species in the Red List of Threatened Species of the International Union for Conservation of Nature. The aims of this research were to survey on the possible regional genetic structure in the southern Gulf of Mexico and to compare genetic status of a landlocked population in Laguna de las Ilusiones (IL) with individuals from localities with no barriers to displacement and breed (open population [OP]). We analyzed 45 manatee skin samples collected from different locations in Tabasco ( n = 38, including 19 from IL), Veracruz ( n = 3), Campeche ( n = 2), and Chiapas ( n = 2). The genomic DNA was isolated and PCR amplifications were performed for each sample using 28 microsatellite loci, previously designed for West Indian manatees and described as polymorphic for this species. Two clusters ( k = 2) were identified by STRUCTURE. The analysis of both a priori populations (IL and OP) indicate that the global values of F ST and R ST ( F ST =0.049, R ST =0.077) were significant. The H E for IL was 0.38 ± 0.03 and for OP was 0.49 ± 0.01. The average number of alleles N A for IL was 2.21 ± 0.09 and for OP was 2.32 ± 0.09. The overall inbreeding coefficient was F IS =−0.013 for analyzed populations. Genetic diversity was low. The IL population had slightly lower genetic diversity compared with OP, which could be explained by isolation of that small group, so conservation plans for IL should be considered as priority.

Tropical Conservation Science