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Ocelots (Leopardus pardalis) are a wide-ranging felid species, occurring from southern United States to northern Argentina. They occupy various habitats and are usually considered the most abundant wild cat species in the Neotropics. However, genetic studies that include free-ranging Mesoamerican ocelots are rare and generally based on small sample sizes. This is the first conservation genetics study on ocelots in Costa Rica and the second one in Mesoamerica that has conducted a genetic assessment of the species at a countrywide scale. We evaluated genetic diversity and population structure of ocelots using 15 microsatellite loci in 28 successfully genotyped individuals from throughout the country. We also compared genetic diversity of Costa Rican ocelots with that of jaguars (Panthera onca) and pumas (Puma concolor) in the country, and with ocelots in Belize. Genetic diversity of ocelots in Costa Rica was relatively high as measured by rarified allelic richness (AR = 5.50 ± 1.36) and expected heterozygosities (HE = 0.79 ± 0.08). We did not detect patterns of genetic substructure, suggesting high levels of gene flow throughout the country and no strong barriers to movement. As expected, genetic diversity of Costa Rican ocelots was higher than co-occurring jaguars and pumas. Additionally, levels of genetic diversity were slightly higher in Costa Rican ocelots when compared with their counterparts in Belize, confirming the south to north decrease in genetic diversity reported in other studies. Our study provides critical baseline information to understand the status of wild ocelot populations in Costa Rica. Future studies on ocelots and other threatened or keystone species should also integrate genetic monitoring and conservation genetics analysis to properly inform management decisions, guarantee their long-term survival, and improve the resilience of ecosystems.

Monitoring the demographics and genetics of reintroduced populations is critical to evaluating reintroduction success, but species ecology and the landscapes that they inhabit often present challenges for accurate assessments. If suitable habitats are restricted to hierarchical dendritic networks, such as river systems, animal movements are typically constrained and may violate assumptions of methods commonly used to estimate demographic parameters. Using genetic detection data collected via fecal sampling at latrines, we demonstrate applicability of the spatial capture–recapture (SCR) network distance function for estimating the size and density of a recently reintroduced North American river otter (Lontra canadensis) population in the Upper Rio Grande River dendritic network in the southwestern United States, and we also evaluated the genetic outcomes of using a small founder group (n = 33 otters) for reintroduction. Estimated population density was 0.23–0.28 otter/km, or 1 otter/3.57–4.35 km, with weak evidence of density increasing with northerly latitude (β = 0.33). Estimated population size was 83–104 total otters in 359 km of riverine dendritic network, which corresponded to average annual exponential population growth of 1.12–1.15/year since reintroduction. Growth was ≥40% lower than most reintroduced river otter populations and strong evidence of a founder effect existed 8–10 years post‐reintroduction, including 13–21% genetic diversity loss, 84%–87% genetic effective population size decline, and rapid divergence from the source population (F ST accumulation = 0.06/generation). Consequently, genetic restoration via translocation of additional otters from other populations may be necessary to mitigate deleterious genetic effects in this small, isolated population. Combined with non‐invasive genetic sampling, the SCR network distance approach is likely widely applicable to demogenetic assessments of both reintroduced and established populations of multiple mustelid species that inhabit aquatic dendritic networks, many of which are regionally or globally imperiled and may warrant reintroduction or augmentation efforts., Monitoring the demographics and genetics of reintroduced populations is critical to evaluating reintroduction success, but species ecology and the landscapes that they inhabit often present challenges for accurate assessments. Using genetic detection data collected via fecal sampling at latrines, we demonstrate applicability of the spatial capture–recapture (SCR) network distance function for estimating the size and density of a recently reintroduced North American river otter (Lontra canadensis) population in the Upper Rio Grande River dendritic network in the southwestern United States, and we also evaluated the genetic outcomes of using a small founder group for reintroduction. Combined with noninvasive genetic sampling, the SCR network distance approach is likely widely applicable to demogenetic assessments of both reintroduced and established populations of multiple mustelid species that inhabit aquatic dendritic networks, many of which are regionally or globally imperiled and may warrant reintroduction or augmentation efforts.

The last known red wolves were captured in southwestern Louisiana and eastern Texas in 1980 to establish a captive breeding population. Before their extirpation, gene flow with coyotes resulted in the persistence of endangered red wolf genetic variation in local coyote populations. We assessed genomic ancestry and morphology of coyotes in southwestern Louisiana. We detected that 38 to 62% of the coyote genomes contained red wolf ancestry acquired in the past 30 years and have an admixture profile similar to that of the canids captured before the extirpation of red wolves. We further documented a positive correlation between ancestry and weight. Our findings highlight the importance of hybrids and admixed genomes as a reservoir of endangered species ancestry for innovative conservation efforts. Together, this work presents an unprecedented system that conservation can leverage to enrich the recovery program of an endangered species.

BackgroundHybridization can be a conservation concern if genomic introgression leads to the loss of an endangered species’ unique genome, or when hybrid offspring are sterile or less fit than their parental species. Yet hybridization can also be an adaptive management tool if rare populations are inbred and have reduced genetic variation, and there is the opportunity to enhance genetic variation through hybridization. The red wolf (Canis rufus) is a critically endangered wolf endemic to the eastern United States, where all extant red wolves are descended from 14 founders which has led to elevated levels of inbreeding over time. Red wolves were considered extirpated from the wild by 1980, but before they disappeared, they interbred with encroaching coyotes creating a genetically admixed population of canids along coastal Texas and Louisiana. In 2018, a genetic study identified individuals on Galveston Island, Texas with significant amounts of red wolf ancestry. We collected 214 samples from Galveston for a more in-depth analysis of this population to identify the amount of red wolf ancestry present and potential mechanisms that support retention of red wolf ancestry on the landscape. ResultsWe identified 23 individual coyotes from Galveston Island and 8 from mainland Texas with greater than 10% red wolf ancestry. Two of those individuals from mainland Texas had greater than 50% red wolf ancestry estimates. Additionally, this population had 6 private alleles that were absent in the North America reference canid populations used in this study, which included 107 southeastern coyotes, 19 captive red wolves, and 38 gray wolves, possibly representing lost red wolf genetic variation. We also identified several individuals on Galveston Island and the mainland of Texas that retained a unique red wolf mitochondrial haplotype present in the red wolf founding population. On Galveston Island, we identified a minimum of four family groups and found coyotes on the island to be highly related, but not genetically depauperate. We did not find clear associations between red wolf ancestry estimates and landscape features, such as open green space or developed areas. ConclusionOur results confirm the presence of substantial red wolf ancestry persisting on Galveston Island and adjacent mainland Texas. This population has the potential to benefit future red wolf conservation efforts through novel reproductive techniques and possibly through de-introgression strategies, with the goals of recovering extinct red wolf genetic variation and reducing inbreeding within the species.

The last red wolves were captured along the Gulf Coast in 1980, where they hybridized with coyote, to establish the captive breeding population. However, red wolf ancestry persists in local coyotes and could be leveraged by genomic innovations to support species persistence. We assessed genomic ancestry and morphology of coyotes in southwestern Louisiana, and find they carried 38-62% red wolf ancestry acquired in the last 30 years, which is enriched on land with minimal coyote hunting. These coyotes were also similar in ancestry to canids captured in the 1970s that initiated the red wolf captive breeding program. Further, we reported that coyotes with higher red wolf ancestry are larger in size. Our findings evidence the importance of hybrids as a reservoir of endangered species ancestry for contemporary conservation efforts. Admixed genomes are at the forefront of innovative solutions, with red wolf survival a prime candidate for this new paradigm.TeaserCoyotes along the American Gulf Coast carry genetic variation critical for the survival of the endangered red wolf species.

Specimens from natural history collections (NHCs) are increasingly being used for genetic studies and can provide information on extinct populations, facilitate comparisons of historical and contemporary populations, produce baseline data before environmental changes, and elucidate patterns of change. Destructive sampling for DNA may be in disagreement with NHC goals of long-term care and maintenance. Differentiating quality among sample sources can direct destructive sampling to the source predicted to yield the highest quality DNA and most reliable data, potentially reducing damage to specimens, laboratory costs, and genotyping errors. We used the kit fox (Vulpes macrotis) as a model species and evaluated the quality and reliability of genetic data obtained from carnivoran specimens via three different sample sources: cranial bones, nasal bones, and toepads. We quantified variation in microsatellite amplification success and genotyping error rates and assessed the reliability of source-specific genic data. Toepads had the highest amplification success rates and lowest genotyping error rates. Shorter loci had higher amplification success and lower allelic dropout rates than longer loci. There were substantial differences in the reliability of resulting multilocus genotypes. Toepads produced the most reliable data, required the fewest replicates, and therefore, had the lowest costs to achieve reliable data. Our results demonstrate that the quality of DNA obtained from specimens varies by sample source and can inform NHCs when evaluating requests for destructive sampling. Our results suggest that prior to large-scale specimen sampling, researchers should conduct pilot studies to differentiate among source-specific data reliability, identify high performing loci, reduce costs of analyses, and minimize destructive sampling.

Information about population demography is crucial for developing and implementing conservation measures. The brown bear in the Gobi desert of southwestern Mongolia (referred to as the Gobi bear) is one of the smallest and most isolated brown bear populations in the world. We conducted genetic sampling (n = 2660 samples collected) using hair corrals around feeding sites at 13 water sources during 2009, 2013, and 2017 to evaluate population size, survival, and population trend. Bears were identified using 13 microsatellite loci and one sex marker. We detected 51 unique individuals (15F and 36M) from our targeted surveys in 2009, 2013, and 2017. Based on capture–mark–recapture robust design, population estimates were 23 (95% CI: 21–32) in 2009, 28 (95% CI: 25–35) in 2013, and 31 (95% CI: 29‐38) individuals in 2017. Spatial capture–recapture analysis suggested abundance was very low (N^ = 27; 95% CI: 22–35), and there was no significant change from 2009 to 2017. The population density was 0.93 bears/1000 km2 (95% CI: 0.74–1.17). Our population estimates suggested a stable population trend. However, the population is still very small, and the sex ratio is skewed toward males, raising concerns for future persistence. Annual survival based on Robust design CMR was 0.85. Low abundance and apparent survival for both sexes in this unhunted population coupled with a skewed sex ratio highlight the need for on‐the‐ground conservation action to conserve this isolated population of bears.

Loss of genetic diversity has serious conservation consequences (e.g., loss of adaptive potential, reduced population viability), but is difficult to evaluate without developing long‐term, multigenerational datasets. Alternatively, historical samples can provide insights into changes in genetic diversity and effective population size (N e). Kit foxes (Vulpes macrotis) are a species of conservation concern across much of their range. In western Utah, kit fox abundance has declined precipitously from historical levels, causing concern about population persistence. We analyzed genetic samples from museum specimens and contemporary scats to evaluate temporal changes in (a) genetic diversity and (b) N e for kit foxes in western Utah, and (c) discuss our findings with respect to population risk and conservation. The N e of kit foxes in western Utah has decreased substantially. When compared to established conservation thresholds for N e (e.g., the 50/500 rule), observed levels suggest the population may be at risk of inbreeding depression and local extinction. In contrast, we found no significant decrease in genetic diversity associated with declining N e. We detected evidence of low levels of immigration into the population and suspect genetic diversity may have been maintained by this previously undescribed gene flow from adjacent populations. Low or intermittent immigration may serve to temper the potential short‐term negative consequences of low N e. We recommend that kit fox conservation efforts focus on evaluating and maintaining landscape connectivity. We demonstrate how historical specimens can provide a baseline of comparison for contemporary populations, highlighting the importance of natural history collections to conservation during a period of declining funding and support.

Admixture resulting from natural dispersal processes can potentially generate novel phenotypic variation that may facilitate persistence in changing environments or result in the loss of population-specific adaptations. Yet, under the US Endangered Species Act, policy is limited for management of individuals whose ancestry includes a protected taxon; therefore, they are generally not protected under the Act. This issue is exemplified by the recently re-established grey wolves of the Pacific Northwest states of Washington and Oregon, USA. This population was likely founded by two phenotypically and genetically distinct wolf ecotypes: Northern Rocky Mountain (NRM) forest and coastal rainforest. The latter is considered potentially threatened in southeast Alaska and thus the source of migrants may affect plans for their protection. To assess the genetic source of the re-established population, we sequenced a ~ 300 bp portion of the mitochondrial control region and ~ 5 Mbp of the nuclear genome. Genetic analysis revealed that the Washington wolves share ancestry with both wolf ecotypes, whereas the Oregon population shares ancestry with NRM forest wolves only. Using ecological niche modelling, we found that the Pacific Northwest states contain environments suitable for each ecotype, with wolf packs established in both environmental types. Continued migration from coastal rainforest and NRM forest source populations may increase the genetic diversity of the Pacific Northwest population. However, this admixed population challenges traditional management regimes given that admixture occurs between an adaptively distinct ecotype and a more abundant reintroduced interior form. Our results emphasize the need for a more precise US policy to address the general problem of admixture in the management of endangered species, subspecies, and distinct population segments.

North American Burbot Lota lota maculosa occupy habitats from deep, cold lakes to prairie streams. Controlled laboratory trials were conducted to evaluate the growth, immune response, and thermal stress physiology of juvenile Burbot subjected to fluctuating, high diel temperatures. Age-0 Burbot from a captive propagation program associated with population restoration in the Kootenai River, Idaho, were tested in temperature cycles that imitated summer conditions observed in regional stream habitats. The diel temperature cycle for the treatment group varied from 18°C at night to 27°C during the day, whereas the control group was held at a constant temperature of 14°C. We evaluated survival, growth, feed efficiency, heat shock protein 70 (hsp70) levels, hsp70 gene expression, serum lysozyme levels, and proximate body composition in samples taken from fish before, during, and at the completion of the trials. Both treatments had high survival and similar growth rates, but the control group gained signi...

The expansion of coyotes (Canis latrans) into the eastern United States has had major consequences for ecological communities and wildlife managers. Despite this, there has been little investigation of the genetics of coyotes across much of this region, especially outside of the northeast. Understanding patterns of genetic structure and interspecific introgression would provide insights into the colonization history of the species, its response to the modern environment, and interactions with other canids. We examined the genetic characteristics of 121 coyotes from the mid-Atlantic states of West Virginia and Virginia by genotyping 17 polymorphic nuclear DNA microsatellite loci. These genotypes were compared with those from other canid populations to evaluate the extent of genetic introgression. We conducted spatial clustering analyses and spatial autocorrelation to assess genetic structure among sampled coyotes. Coyotes across the 2 states had high genetic diversity, and we found no evidence of genetic structure. Six to sixteen percent of individuals displayed some evidence of genetic introgression from other species depending on the method and criteria used, but the population possessed predominantly coyote ancestry. Our findings suggested introgression from other canid populations has played less of a role in shaping the genetic character of coyotes in these states compared with populations closer to the Canadian border. Coyotes appear to display a panmictic population structure despite high habitat heterogeneity and heavy human influence in the spatial environment, underscoring the adaptability of the species.

Summary Although phytophagous insects can vary genetically in host use and exhibit long-range movements, the combined implications of these phenomena for pest management have received limited attention. To address this, we surveyed the genetic diversity of pea aphid Acyrthosiphon pisum using twelve microsatellite loci and assessed host association patterns and annual movement from a putative source region (Columbia River Basin) to the Palouse region of northern Idaho and western Washington, where the aphid is a pest of pea Pisum sativum. A total of 320 identified unique genotypes clustered into four genetic groups, with two host plant associations: alfalfa Medicago sativa (three genetic groups), and pea Pisum sativum and vetch Vicia villosa (one genetic group). All four genetic groups occurred in the Columbia River Basin and in migrant aphids collected in pan traps during spring colonization in the Palouse during 2 years of this study. Patterns of group arrival on the Palouse were spatially structured early in the season, consistent with differing migration patterns. Despite genetic diversity of migrants, a single genetic group became predominant in pea crops each year. Clonal laboratory colonies of pea aphids established from field-collected specimens and representing two predominant genetic groups exhibited reciprocal performance trade-offs, with alfalfa being a poor host for a pea-associated aphid genotype and vice versa. Synthesis and applications. Annual spring migrants of pea aphids in the pea production region of the Palouse are genetically diverse, with different host plant affinities consistent with origination from source populations in the Columbia River Basin. As the season progresses, a single genetic group adapted to pea becomes predominant in the crop. Management of pea aphid in the Palouse will be improved by monitoring the temporal and spatial variation of specific genetic groups of the aphid arriving as immigrants during each crop season, providing this information to producers and adjusting estimates of risk of crop damage accordingly. The principle could apply to other pest species with host-adapted populations that colonize crops on an annual basis.

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Knowledge of genetic diversity and population structure is critical for conservation and management planning at the population level within a species’ range. Many brown bear populations in Central Asia are small and geographically isolated, yet their phylogeographic relationships, genetic diversity, and contemporary connectivity are poorly understood. To address this knowledge gap, we collected brown bear samples from the Gobi Desert (n = 2360), Altai, Sayan, Khentii, and Ikh Khyangan mountains of Mongolia (n = 79), and Deosai National Park in the Himalayan Mountain Range of Pakistan (n = 5) and generated 927 base pairs of mitochondrial DNA (mtDNA) sequence data and genotypes at 13 nuclear DNA microsatellite loci. We documented high levels of mtDNA and nDNA diversity in the brown bear populations of northern Mongolia (Altai, Sayan, Buteeliin nuruu and Khentii), but substantially lower diversity in brown bear populations in the Gobi Desert and Himalayas of Pakistan. We detected 3 brown bear mtDNA phylogeographic groups among bears of the region, with clade 3a1 in Sayan, Khentii, and Buteeliin nuruu mountains, clade 3b in Altai, Sayan, Buteeliin nuruu, Khentii, and Ikh Khyangan, and clade 6 in Gobi and Pakistan. Our results also clarified the phylogenetic relationships and divergence times with other brown bear mtDNA clades around the world. The nDNA genetic structure analyses revealed distinctiveness of Gobi bears and different population subdivisions compared to mtDNA results. For example, genetic distance for nDNA microsatellite loci between the bears in Gobi and Altai (FST = 0.147) was less than that of the Gobi and Pakistan (FST = 0.308) suggesting more recent male-mediated nuclear gene flow between Gobi and Altai than between Gobi and the Pakistan bears. Our results provide valuable information for conservation and management of bears in this understudied region of Central Asia and highlight the need for special protection and additional research on Gobi brown bears.

An understanding of a species’ diet is required to make sound conservation and management decisions. Traditionally, morphological analyses of undigested hard parts from food items remaining in scats have been used to assess diets. More recently, molecular analyses of scats have been used to identify plant and prey species’ DNA, but no studies have compared morphological and molecular diet analyses for large, terrestrial carnivores. We used molecular tools to determine the percentage of black bear and coyote scats that contained 3 common prey species (caribou, moose, and snowshoe hares) in Newfoundland and compared the results to a traditional morphological analysis. We found that a ranking of relative prey frequencies was consistent between the 2 methods, but molecular methods tended to detect prey species in a greater percentage of scats for all prey species. However, there were individual scats in which a prey species was detected by morphological methods only, and we provide evidence that molecular methods could result in false negatives if prey DNA is not uniformly distributed throughout a scat or as a result of PCR inconsistency. We also found that the per sample cost comparison between morphological and molecular analyses was dependent upon whether or not a molecular test was needed to identify scats to the predator species, the cost of developing molecular methods, and the number of samples being processed. We recommend that controlled feeding studies be performed to validate molecular methods and investigate the utility of molecular techniques to estimate the proportions of food items consumed.

The interaction between brown bears (

Hybridization presents a unique challenge for conservation biologists and managers. While hybridization is an important evolutionary process, hybridization is also a threat formany native species. The endangered species recovery effort for the red wolf Canis rufus is a classic system for understanding and addressing the challenges of hybridization. From 1987?1993, 63 red wolves were released from captivity in eastern North Carolina, USA, to establish a free-ranging, non-essential experimental population. By 1999, managers recognized hybridization with invasive coyotes Canis latrans was the single greatest threat to successful recovery, and an adaptive management plan was adopted with innovative approaches for managing the threat of hybridization. Here we review the application and results of the adaptive management efforts from 1993 to 2013 by comparing: (1) the numbers of wolves, coyotes, and hybrids captured, (2) the numbers of territorial social groups with presumed breeding capabilities, (3) the number of red wolf and hybrid litters documented each year and (4) the degree of coyote introgression into the wild red wolf gene pool. We documented substantial increases in the number of known red wolves and red wolf social groups from 1987–2004 followed by a plateau and slight decline by 2013.The number of red wolf litters exceeded hybrid litters each year and the proportion of hybrid litters per year averaged 21%. The genetic composition of the wild red wolf population is estimated to include < 4% coyote ancestry from recent introgression since reintroduction. We conclude that the adaptive management plan was effective at reducing the introgression of coyote genes into the red wolf population, but population recovery of red wolves will require continuation of the current management plan, or alternative approaches, for the foreseeable future. More broadly, we discuss the lessons learned from red wolf adaptive management that could assist other endangered species recovery efforts facing the challenge of minimizing hybridization.

Endangered red wolves (Canis rufus) receive intense conservation efforts in the United States, and to date, population recovery has been challenged by hybridization with closely related coyotes (C. latrans) and illegal human-caused mortality. Ongoing review of the red wolf program in the single recovery area in North Carolina prompted us to compare demography (survival, recruitment) and cause of death of red wolves and coyotes/hybrids. In most respects, canids had similar demographic rates, although sterilization was effective in controlling coyote reproduction. Comparison of previous (1999-2007) to contemporary (2009-2014) causes of death revealed that shooting mortality consistently accounted for ∼25% of wolf mortality. As evidenced by the lack of coyote deaths from strife with wolves, and stationary/declining wolf numbers during the last 15 years, current conditions are inadequate to establish a viable self-sustaining wolf population. Accordingly, the program review should determine whether: (1) banning coyote hunting will sufficiently benefit wolf survival or recruitment; (2) the wolf population should be considered conservation-reliant under revised recovery goals; or (3) the recovery program in North Carolina should be abandoned.

The development of high-throughput sequencing technologies is dramatically increasing the use of single nucleotide polymorphisms (SNPs) across the field of genetics, but most parentage studies of wild populations still rely on microsatellites. We developed a bioinformatic pipeline for identifying SNP panels that are informative for parentage analysis from restriction site-associated DNA sequencing (RADseq) data. This pipeline includes options for analysis with or without a reference genome, and provides methods to maximize genotyping accuracy and select sets of unlinked loci that have high statistical power. We test this pipeline on small populations of Mexican gray wolf and bighorn sheep, for which parentage analyses are expected to be challenging due to low genetic diversity and the presence of many closely related individuals. We compare the results of parentage analysis across SNP panels generated with or without the use of a reference genome, and between SNPs and microsatellites. For Mexican gray wolf, we conducted parentage analyses for 30 pups from a single cohort where samples were available from 64% of possible mothers and 53% of possible fathers, and the accuracy of parentage assignments could be estimated because true identities of parents were known a priori based on field data. For bighorn sheep, we conducted maternity analyses for 39 lambs from five cohorts where 77% of possible mothers were sampled, but true identities of parents were unknown. Analyses with and without a reference genome produced SNP panels with ≥95% parentage assignment accuracy for Mexican gray wolf, outperforming microsatellites at 78% accuracy. Maternity assignments were completely consistent across all SNP panels for the bighorn sheep, and were 74.4% consistent with assignments from microsatellites. Accuracy and consistency of parentage analysis were not reduced when using as few as 284 SNPs for Mexican gray wolf and 142 SNPs for bighorn sheep, indicating our pipeline can be used to develop SNP genotyping assays for parentage analysis with relatively small numbers of loci.

In a recent article, vonHoldt and colleagues ( 1 ) use whole-genome sequence data to address the relationships and evolutionary origins of several North American canid taxa. The authors conclude that two taxa, the red wolf ( Canis rufus ) and the eastern wolf ( Canis lycaon ), are not long-diverged lineages as other researchers have concluded from genetic ( 2 – 7 ) and fossil evidence ( 8 , 9 ) but are, rather, populations resulting from recent admixture between gray wolves ( Canis lupus ) and coyotes ( Canis latrans ). They use this conclusion to argue that endangered species policy should better account for hybridization and admixture. We agree that hybridization and admixture are widespread in the natural world and that the conservation policy needs to be updated to account for this biological reality. However, we strongly disagree with their conclusion that red and eastern wolves are of recent hybrid origin and we conclude that their analysis does not actually test the hypothesis of a recent hybrid origin. Their data are consistent with multiple hypotheses for the origins of red and eastern wolves, including relatively old origins of these taxa. Furthermore, we argue that their data do not support “a lack of unique ancestry in red and eastern wolves” ( 1 ); rather, substantial evidence still supports the conclusion that red and eastern wolves represent genetically distinct taxa among North American canids. Below, we detail this perspective and argue for further analyses that would directly test competing hypotheses for the evolutionary origins of these groups. Analysis of whole-genome sequence data necessarily relies on a relatively small number of individual samples that are taken to be representative of larger populations or taxa. In this case, the choice of representative samples is problematic in several respects. First, of the three individuals chosen as representative nonadmixed coyotes, …

Estimating densities of cougar (Puma concolor) is important for managing cougars and their prey but remains challenging because of cougar's elusive and solitary behavior. To evaluate a non-invasive, genetic capture–recapture method to estimate cougar population size and density, we surveyed a 220-km2 area using conservation detection dogs trained to locate scat over a 4-week sampling period in northeast Oregon. We collected 272 scat samples and conducted DNA analysis on 249 samples from which we determined individual identification from 73 samples that represented 21 cougars (9 males and 12 females). We evaluated 4 models to estimate cougar densities: Huggins closed population capture–recapture (Huggins), CAPWIRE, multiple detections with Poisson (MDP), and spatially explicit capture–recapture (SECR). Population estimates for cougars using our study area were 26 (95% CI = 22–35, 9 males and 17 females) from Huggins models, 24 (95% CI = 21–30, 9 males and 15 females) from CAPWIRE, and 27 (95% CI = 24–42, 9 males and 18 females) from the MDP model. We accounted for the edge effect in density estimates caused by individuals whose home ranges included only a portion of the survey grid by buffering the study area using the mean home range radius of 8 cougars equipped with global positioning system collars on or near the study area. We estimated densities of 4.6 cougars/100 km2 (95% CI = 3.8–8.3) for the Huggins model, 4.8 cougars/100 km2 (95% CI = 4.2–7.8) for the MDP model, 4.2 cougars/100 km2 (95% CI = 3.3–5.3) for the CAPWIRE model, and 5.0 cougars/100 km2 (95% CI = 3.2–7.7) for the SECR model. Our results suggested estimating cougar densities using scat detection dogs could be feasible at a broader scale with less effort than other methods currently being used. © 2014 The Wildlife Society.

While genetic rescue is known to benefit population viability, the duration of that benefit is poorly understood. We document what appears to be the waning benefit of genetic rescue after approximately 2–3 generations for the wolf population in Isle Royale National Park. The fitness benefit of genetic rescue declined because of inbreeding and population abundance declined when the inbred individuals exhibited low reproduction and survival. Only detailed studies of other cases will reveal what aspects of these dynamics represent general features of genetic rescue. We also present evidence indicating that numerous past immigration events have likely gone undetected. This finding is of particular significance because the Isle Royale wolf population has maintained good population viability for decades even though it was small and thought to be isolated from the mainland population of wolves. Past gene flow also suggests that human-assisted gene flow is necessary to conserve the ecosystem services associated with predation, since climate warming has reduced the frequency of ice bridges and with it the only opportunity for unassisted gene flow.

The pygmy rabbit Brachylagus idahoensis is a sagebrush-obligate species of conservation concern that occurs in the Great Basin and adjacent intermountain areas in the western United States. The species is not known to occur in Colorado, despite proximity to existing populations of pygmy rabbits in Wyoming. We provide the first documentation of the pygmy rabbit in Colorado. Fecal pellets diagnostic of pygmy rabbits were collected in the Vermillion Bluffs area of northwestern Colorado. Samples from 16 pellet clusters were collected for species identification via genetic analyses, and we were able to extract and amplify sufficient DNA from 7 of 16 pellet samples. All seven samples were identified as originating from pygmy rabbits based on a species-specific mitochondrial DNA fragment analysis test. To verify species identification, we also sequenced 225 base pairs of the mitochondrial DNA cytochrome b region from all seven pellet samples. Presence of pygmy rabbits was confirmed from three locations separated by 2.4–7.7 km and pellets represented both adult and juvenile rabbits. Based on the sparseness of burrows in the area, density of pygmy rabbits in the area likely is low; however, systematic surveys by trained observers are needed to delineate the range and density of this species in Colorado. Given the conservation concern for pygmy rabbits across their current range, the newly confirmed presence of this species suggests that assessment of their conservation status in Colorado is warranted.

Sex-specific senescence has been construed as a function of mating system and differential investment in parental care, with males exhibiting low parental investment predicted to have more rapid senescence due to costly reproductive behavior. In monogamous mating systems, however, where parental investment may be more evenly distributed, rates of senescence are predicted to be more equivalent between the sexes than in polygynous mating systems. While many polygynous species do appear to support this pattern, evidence from monogamous species, particularly mammals, is scarce. Wolves are an excellent system with which to test this hypothesis, as they exhibit both monogamy and cooperative breeding, where parental investment is distributed across both breeders and non-breeders of both sexes. We examined patterns of age-specific reproduction in red wolves and red wolf-coyote hybrids. We found no evidence of decline in pup production with age in male wolves; among females, the production appeared better explained by hybrid status and age at first reproduction than age per se. Remarkably, however, there was strong evidence of sex differences in pup recruitment, with males, but not females, showing a steep decline in recruitment with age. Combined with previous work on aging in captive wolves, our findings contribute not only to the current understanding of the relationship between mating system and senescence but also to the plasticity of aging and the dynamics of female mate choice in the wild. In this study, we test for signs of reproductive aging in red wolves using a 20-year dataset documenting age-specific reproduction in the wild. We find evidence that reproductive aging in males is evident for number of pups recruited, though not for number of pups born; we find no evidence for reproductive aging at either stage in wild female wolves. This sex difference stands in contrast to the general prediction that patterns of aging would be more monomorphic among monogamous species. Furthermore, this is especially of interest given that red wolves are cooperative breeders, with costs of reproduction spread across family groups. In general, this study constitutes a valuable contribution to an emerging literature geared at understanding patterns of aging in the context of diverse social and ecological variables.

Bayesian clustering methods have emerged as a popular tool for assessing hybridization using genetic markers. Simulation studies have shown these methods perform well under certain conditions; however, these methods have not been evaluated using empirical data sets with individuals of known ancestry. We evaluated the performance of two clustering programs, baps and structure, with genetic data from a reintroduced red wolf (Canis rufus) population in North Carolina, USA. Red wolves hybridize with coyotes (C. latrans), and a single hybridization event resulted in introgression of coyote genes into the red wolf population. A detailed pedigree has been reconstructed for the wild red wolf population that includes individuals of 50-100% red wolf ancestry, providing an ideal case study for evaluating the ability of these methods to estimate admixture. Using 17 microsatellite loci, we tested the programs using different training set compositions and varying numbers of loci. structure was more likely than baps to detect an admixed genotype and correctly estimate an individual's true ancestry composition. However, structure was more likely to misclassify a pure individual as a hybrid. Both programs were outperformed by a maximum-likelihood-based test designed specifically for this system, which never misclassified a hybrid (50-75% red wolf) as a red wolf or vice versa. Training set composition and the number of loci both had an impact on accuracy but their relative importance varied depending on the program. Our findings demonstrate the importance of evaluating methods used for detecting admixture in the context of endangered species management.

Among cooperatively breeding species, delayed dispersal provides an opportunity for older siblings to help rear young. There is considerable variation in the extent to which cooperative breeders delay dispersal, however, and while delayed dispersal may be a consequence of adaptive evolution favouring family living, evidence for a genetic basis is lacking. Furthermore, the extent to which dispersal behaviour itself is under direct selection, or evolves indirectly in response to selection on correlated life-history traits, remains unclear. We tested for evidence of inter- and intraspecific quantitative genetic variation in age at first dispersal in a reintroduced population of the red wolf, Canis rufus , that naturally hybridizes with the coyote, Canis latrans , which delays dispersal to a lesser extent than the red wolf. Using analyses based on individual relationships to population founders and sire–offspring regressions, we found evidence for intraspecific genetic variation in dispersal age in male, but not female, red wolves. We also found evidence for interspecific genetic variation, in that red wolves had later dispersal age than coyote–red wolf hybrids. Intraspecific variation in dispersal age was not explained by variation in social and environmental factors nor alternative targets of selection such as body mass or age at first reproduction. Thus, selection may act directly on dispersal-related behaviours. However, life-history traits may continue to be influential in creating interspecific differences in dispersal age, as hybrids also showed earlier reproduction. Our findings suggest that evolution of delayed dispersal may be due to both direct and indirect selection on existing quantitative genetic variation, and that genetic and environmental mechanisms underlying dispersal may differ between the sexes.

Client religious and spiritual practices have recently emerged as beneficial to both mental and physical health. However, graduates of counseling programs indicate that they have not been adequately trained to address religious and spiritual issues with clients. This exploratory study focused on the perceptions of counselors-in-training regarding what messages they received about these issues during their training programs, as well as their potential behaviors when working with clients. Results indicated that potential behaviors are not always consistent with what they are taught. Implications for training and future research are discussed.

For cooperatively breeding groups composed of close relatives, whether and how a group avoids inbreeding are questions of key evolutionary and conservation importance. A number of strategies for inbreeding avoidance may be employed by cooperative breeders, including extrapair reproduction, reproductive suppression, and juvenile dispersal. However, population-wide information on the prevalence of different strategies is difficult to obtain. We investigated the prevalence of inbreeding and potential mechanisms for inbreeding avoidance in a reintroduced population of the red wolf. Using long-term data on individuals of known pedigree, we determined that inbreeding among first-degree relatives was rare. Potential mechanisms for inbreeding avoidance included low levels of philopatric reproduction in spite of delayed dispersal, and reproductive suppression prior to dispersal. Inbreeding avoidance among siblings may have been further facilitated by independent dispersal trajectories, as many young wolves spent time alone or in small nonbreeding packs composed of unrelated individuals. The dominant pattern of breeding-pair formation involved the union of 2 unrelated individuals in a new home range. Replacement of 1 or both members of an existing breeding pair involved new immigrants to a pack or, in a small number of cases, ascendance of either resident offspring or adopted pack members to vacant breeding positions. Extrapair reproduction was rare, suggesting that it was not a major mechanism for outbreeding. We conclude that there are several prevalent behavioral strategies within the red wolf population that may work together to minimize inbreeding and any associated fitness costs, helping make cooperative breeding an evolutionarily viable strategy. Key wor ds: Canis r ufus, competition, lone wolves, monogamy, nonbreeding packs, pair formation. [Behav Ecol]

Mating with close kin can lead to inbreeding depression through the expression of recessive deleterious alleles and loss of heterozygosity. Mate selection may be affected by kin encounter rate, and inbreeding avoidance may not be uniform but associated with age and social system. Specifically, selection for kin recognition and inbreeding avoidance may be more developed in species that live in family groups or breed cooperatively. To test this hypothesis, we compared kin encounter rate and the proportion of related breeding pairs in noninbred and highly inbred canid populations. The chance of randomly encountering a full sib ranged between 1–8% and 20–22% in noninbred and inbred canid populations, respectively. We show that regardless of encounter rate, outside natal groups mates were selected independent of relatedness. Within natal groups, there was a significant avoidance of mating with a relative. Lack of discrimination against mating with close relatives outside packs suggests that the rate of inbreeding in canids is related to the proximity of close relatives, which could explain the high degree of inbreeding depression observed in some populations. The idea that kin encounter rate and social organization can explain the lack of inbreeding avoidance in some species is intriguing and may have implications for the management of populations at risk.

The pygmy rabbit (Brachylagus idahoensis) is a small lagomorph of the western United States that specializes in sagebrush (Artemisia spp.) habitat. Intensive habitat loss and modification have increased the vulnerability of pygmy rabbit populations, but the current geographic distribution and population status remain unclear. To aid in detection and population monitoring, we developed a species identification test that uses mitochondrial DNA species-specific primers to distinguish among six sympatric lagomorph species using DNA isolated from faecal pellets. Applying this test, we successfully identified the species of origin for all pellet samples that produced a positive PCR result (77% of 283 pellets collected). Pellets collected during the winter (December-February) had higher PCR success rate (93%) than pellets collected at other times of the year (72%). This test, using non-invasive genetic sampling of faecal pellets, provides an efficient method for assessing site occupancy and distribution of pygmy rabbits and other lagomorphs across large geographic areas.

Genetic rescue, in which the introduction of one or more unrelated individuals into an inbred population results in the reduction of detrimental genetic effects and an increase in one or more vital rates, is a potentially important management tool for mitigating adverse effects of inbreeding. We used molecular techniques to document the consequences of a male wolf (Canis lupus) that immigrated, on its own, across Lake Superior ice to the small, inbred wolf population in Isle Royale National Park. The immigrant's fitness so exceeded that of native wolves that within 2.5 generations, he was related to every individual in the population and his ancestry constituted 56 per cent of the population, resulting in a selective sweep of the total genome. In other words, all the male ancestry (50% of the total ancestry) descended from this immigrant, plus 6 per cent owing to the success of some of his inbred offspring. The immigration event occurred in an environment where space was limiting (i.e. packs occupied all available territories) and during a time when environmental conditions had deteriorated (i.e. wolves' prey declined). These conditions probably explain why the immigration event did not obviously improve the population's demography (e.g. increased population numbers or growth rate). Our results show that the beneficial effects of gene flow may be substantial and quickly manifest, short-lived under some circumstances, and how the demographic benefits of genetic rescue might be masked by environmental conditions.

The existence of cooperative breeding in diverse animal taxa has inspired much interest in what nonbreeding helpers gain from participation in rearing nondescendent young. A major theoretical explanation for this phenomenon has revolved around the notion of inclusive fitness, where delayed dispersers in a family-based group gain indirect fitness benefits by fostering the viability of close relatives. There is potential, however, for direct fitness benefits in delayed dispersal itself. We explored the relationship between delayed dispersal and lifetime fitness in a reintroduced population of the cooperatively breeding red wolf, Canis rufus, which exhibits delayed dispersal but few opportunities to breed in the natal pack. We present evidence that male wolves that delayed dispersal to later ages had lower mortality risk from natural and anthropogenic sources combined and increased probability of becoming reproductive in their lifetimes. Furthermore, delayed dispersal did not result in delayed age at first reproduction. For females, however, the relative costs and benefits of delaying dispersal to later ages were more complex. In general, we provide evidence that there are direct fitness benefits to delaying dispersal in red wolves even in the absence of reproductive opportunities in the natal pack. Thus, we lend support to the hypothesis that direct fitness benefits may in themselves be sufficient to facilitate the evolution of delayed dispersal requisite to cooperatively breeding social systems. Key words: age at first dispersal, age at first reproduction, canids, cooperative breeding, lifetime reproductive probability, lifetime reproductive success, survival. [Behav Ecol]

Spatial capture-recapture models can produce unbiased estimates of population density, but sparse detection data often plague studies of social and territorial carnivores. Integrating multiple types of detection data can improve estimation of the spatial scale parameter (sigma), activity center locations, and density. Noninvasive genetic sampling is effective for detecting carnivores, but social structure and territoriality could cause differential detectability among population cohorts for different detection methods. Using three observation models, we evaluated the integration of genetic detection data from noninvasive hair and scat sampling of the social and territorial coyote (Canis latrans). Although precision of estimated density was improved, particularly if sharing sigma between detection methods was appropriate, posterior probabilities of sigma and posterior predictive checks supported different sigma for hair and scat observation models. The resulting spatial capture-recapture model described a scenario in which scat-detected individuals lived on and around scat transects, whereas hair-detected individuals had larger sigma and mostly lived off of the detector array, leaving hair but not scat samples. A more supported interpretation is that individual heterogeneity in baseline detection rates (lambda(0)) was inconsistent between detection methods, such that each method disproportionately detected different population cohorts. These findings can be attributed to the sociality and territoriality of canids: Residents may be more likely to strategically mark territories via defecation (scat deposition), and transients may be more likely to exhibit rubbing (hair deposition) to increase mate attraction. Although this suggests that reliance on only one detection method may underestimate population density, integrating multiple sources of genetic detection data may be problematic for social and territorial carnivores. These data are typically sparse, modeling individual heterogeneity in lambda(0) and/or sigma with sparse data is difficult, and positive bias can be introduced in density estimates if individual heterogeneity in detection parameters that is inconsistent between detection methods is not appropriately modeled. Previous suggestions for assessing parameter consistency of sigma between detection methods using Bayesian model selection algorithms could be confounded by individual heterogeneity in lambda(0) in noninvasive detection data. We demonstrate the usefulness of augmenting those approaches with calibrated posterior predictive checks and plots of the posterior density of activity centers for key individuals. U.S. Fish and Wildlife Service [F15AC01292] Louisiana Department of Wildlife and Fisheries University of Kentucky Department of Forestry and Natural Resources This study was funded by Cooperative Agreement Award #F15AC01292 from the U.S. Fish and Wildlife Service. Supplemental funds were provided by Louisiana Department of Wildlife and Fisheries and the University of Kentucky Department of Forestry and Natural Resources. We thank Emily Carrollo and Andrea Petrullo for assistance with data collection and Meaghan Clark and Michelle Keyes for assistance with laboratory genetic analysis. We are grateful for the support from Robert Gosnell, Leopoldo Miranda, and Billy Leonard. We also thank staff at Southwest Louisiana National Wildlife Refuge Complex, Louisiana Ecological Services Office, and Louisiana Department of Wildlife and Fisheries for providing logistical support. Author contributions: SMM conceived and designed the study, acquired funding, collected data, analyzed capture-recapture data, and led writing; BCA developed MCMC algorithms and analyzed capture-recapture data; JRA and LPW conducted laboratory genetic analysis; and JJC contributed to study design and acquired supplemental funding. All authors participated in discussion of findings, writing and reviewing of the manuscript, and provided final approval for publication. To the best of our knowledge, no conflict of interest, financial, relational, or other, exists.

The authors note that although the Diagnostic and Statistical Manual of Mental Disorders (4th ed., text rev.; American Psychiatric Association, 2000) provides a useful tool for assessment and treatment planning, there has been debate over the lack of attention to issues of diversity. The elements of this debate are presented, along with arguments for inclusion of client strengths and resources as alternatives for diagnosis alone.

Utilizing creative techniques within the counseling session makes the content come alive for both counselor and client. When combined with a solid theoretical foundation, creative metaphors provide a powerful tool for empowering clients and helping them sustain change. This article describes a creative way to use the Winnie the Pooh characters to illustrate the Transactional Analysis ego states. Case examples are included.

Understanding the genetics of a population is a critical component of developing conservation strategies. We used archived tissue samples from golden eagles (Aquila chrysaetos canadensis) in three geographic regions of western North America to conduct a preliminary study of the genetics of the North American subspecies, and to provide data for United States Fish and Wildlife Service (USFWS) decision-making for golden eagle management. We used a combination of mitochondrial DNA (mtDNA) D-loop sequences and 16 nuclear DNA (nDNA) microsatellite loci to investigate the extent of gene flow among our sampling areas in Idaho, California and Alaska and to determine if we could distinguish birds from the different geographic regions based on their genetic profiles. Our results indicate high genetic diversity, low genetic structure and high connectivity. Nuclear DNA Fst values between Idaho and California were low but significantly different from zero (0.026). Bayesian clustering methods indicated a single population, and we were unable to distinguish summer breeding residents from different regions. Results of the mtDNA AMOVA showed that most of the haplotype variation (97%) was within the geographic populations while 3% variation was partitioned among them. One haplotype was common to all three areas. One region-specific haplotype was detected in California and one in Idaho, but additional sampling is required to determine if these haplotypes are unique to those geographic areas or a sampling artifact. We discuss potential sources of the high gene flow for this species including natal and breeding dispersal, floaters, and changes in migratory behavior as a result of environmental factors such as climate change and habitat alteration. Our preliminary findings can help inform the USFWS in development of golden eagle management strategies and provide a basis for additional research into the complex dynamics of the North American subspecies.

Hybridization with coyotes ( Canis latrans ) continues to threaten the recovery of endangered red wolves ( Canis rufus ) in North Carolina and requires the development of new strategies to detect and remove coyotes and hybrids. Here, we combine a spatially targeted faecal collection strategy with a previously published reference genotype data filtering method and a genetic test for coyote ancestry to screen portions of the red wolf experimental population area for the presence of nonred wolf canids. We also test the accuracy of our maximum-likelihood assignment test for identifying hybrid individuals using eight microsatellite loci instead of the original 18 loci and compare its performance to the Bayesian approach implemented in NEWHYBRIDS . We obtained faecal DNA genotypes for 89 samples, 73 of which were matched to 23 known individuals. The performance of two sampling strategies — comprehensive sweep and opportunistic spot-check was evaluated. The opportunistic spot-check sampling strategy required less effort than the comprehensive sweep sampling strategy but identified fewer individuals. Six hybrids or coyotes were detected and five of these individuals were subsequently captured and removed from the population. The accuracy and power of the genetic test for coyote ancestry is decreased when using eight loci; however, nonred wolf canids are identified with high frequency. This combination of molecular and traditional field-based approaches has great potential for addressing the challenge of hybridization in other species and ecosystems.