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Non-invasive DNA sampling to identify and enumerate species is critical to population monitoring and for developing effective management strategies. However, individual DNA identification is often limited by degraded and low template DNA (LT-DNA) that routinely yields partial profiles prone to technical artifacts, thus limiting their utility/reliability. Massively parallel, genotyping-by-sequencing (GBS) assays present an opportunity to amplify not only a large suite of molecular markers simultaneously, providing higher resolution to identify individuals, but also higher levels of sequence redundancy to enable quality metric evaluations of profiles from LT-DNA. Taxidea taxus jacksoni is an endangered badger subspecies in Canada, with low levels of genetic diversity, complicating individual identifications from closely related DNA sequences. Challenges arise from the small number of hairs collected from snag traps set in badger burrows that rarely provide full profiles. We designed a GBS assay to obtain microsatellite profiles compatible with pre-existing databases generated with conventional capillary electrophoresis (CE) genotyping. We assessed the assay’s reproducibility via a dilution series to mimic LT-DNA and tested if the assay produced similar CE-generated results. While GBS offers the potential to genotype large numbers of individuals and markers at the same time, we found low concordance between GBS- and CE-based genotypes from DNA templates

Ranaviruses have been associated with chelonian mortality. In Canada, the first two cases of ranavirus were detected in turtles in 2018 in Ontario, although a subsequent survey of its prevalence failed to detect additional positive cases. To confirm the prevalence of ranavirus in turtles in Ontario, we used a more sensitive method to investigate if lower level persistent infection was present in the population. Here we report results via a combination of qPCR, PCR, Sanger sequencing and genome sequencing from turtles from across Ontario, with no clinical signs of illness. We found 2 positives with high viral load and 5 positives with low viral load. Histopathology found subtle histological changes. DNA sequences identified two types of frog virus 3 (FV3), and genome sequencing identified a ranavirus similar to wild-type FV3. Our results show that the virus has been present in Ontario’s turtles as subclinical infections.

Muskoxen (

Skin is a key aspect of the immune system in the defence against pathogens. Skin pH regulates the activity of enzymes produced both by hosts and by microbes on host skin, thus implicating pH in disease susceptibility. Skin pH varies inter- and intra-specifically and is influenced by a variety of intrinsic and extrinsic variables. Increased skin alkalinity is associated with a predisposition to cutaneous infections in humans and dogs, and inter-specific and inter-individual variation in skin pH is implicated in differential susceptibility to some skin diseases. The cutaneous pH of bats has not been characterized but is postulated to play a role in susceptibility to white-nose syndrome (WNS), a fungal infection that has decimated several Nearctic bat species. We used non-invasive probes to measure the pH of bat flight membranes in five species with differing susceptibility to WNS. Skin pH ranged from 4.67 to 8.59 and varied among bat species, geographic locations, body parts, age classes, sexes and seasons. Wild Eptesicus fuscus were consistently more acidic than wild Myotis lucifugus, Myotis leibii and Perimyotis subflavus. Juvenile bats had more acidic skin than adults during maternity season but did not differ during swarming. Male M. lucifugus were more acidic than females during maternity season, yet this trend reversed during swarming. Bat skin was more acidic in summer compared to winter, a pattern also reported in humans. Skin pH was more acidic in captive than wild E. fuscus, suggesting environmental impacts on skin pH. The pH of roosting substrates affects skin pH in captive bats and may partially explain seasonal patterns in wild bats that use different roost types across seasons. Future research on the influence of pH on microbial pathogenic factors and skin barrier function may provide valuable insights on new therapeutic targets for treating bat skin conditions.

There is a recognized disconnect in priority and synergy between academic and practitioners in forensic science. In this work, we personally reflect on our experiences in conducting research studies that directly involve academic and practitioner stakeholders. We believe, amongst others, that this "gap" can be mitigated through regular and productive communication. We also emphasize the need to create stronger and national research strategies which identifies the current and pressing needs of enforcement officials in order to bring these needs directly to academia. As part of this, researchers should actively seek to make sure what they study will be relevant within the discipline. Our reflection is geared on direct feedback from an entomological study in large scale sampling of blowflies and workshops in bloodstain pattern analysis using a forensic blood substitute.

The Canadian Arctic is an extreme environment with low floral and faunal diversity characterized by major seasonal shifts in temperature, moisture, and daylight. Muskoxen (Ovibos moschatus) are one of few large herbivores able to survive this harsh environment. Microbiome research of the gastrointestinal tract may hold clues as to how muskoxen exist in the Arctic, but also how this species may respond to rapid environmental changes. In this study, we investigated the effects of season (spring/summer/winter), year (2007–2016), and host genetic structure on population‐level microbiome variation in muskoxen from the Canadian Arctic. We utilized 16S rRNA gene sequencing to characterize the fecal microbial communities of 78 male muskoxen encompassing two population genetic clusters. These clusters are defined by Arctic Mainland and Island populations, including the following: (a) two mainland sampling locations of the Northwest Territories and Nunavut and (b) four locations of Victoria Island. Between these geographic populations, we found that differences in the microbiome reflected host‐associated genetic cluster with evidence of migration. Within populations, seasonality influenced bacterial diversity with no significant differences between years of sampling. We found evidence of pathogenic bacteria, with significantly higher presence in mainland samples. Our findings demonstrate the effects of seasonality and the role of host population‐level structure in driving fecal microbiome differences in a large Arctic mammal., We analyzed the microbiome and nuclear microsatellites from muskoxen from two genetically distinct Canadian populations (Mainland and Victoria Island). Our results showed differences in the microbiome reflected host‐associated genetic cluster with evidence of migration between these two regions. We found evidence of pathogenic bacteria, with significantly higher presence in Mainland samples.

Low genetic diversity is associated with low fitness and evolutionary potential, yet the demographic and life-history traits of some species contribute to low genetic diversity, without empirical evidence of negative impacts on fitness. Modelling past and future trajectories of genetic diversity under different demographic scenarios can provide insight into how genetic variation might impact population fitness. The muskox is an Arctic species that has undergone multiple population bottlenecks and, although populations have rebounded repeatedly, two large populations have recently declined by > 50%. It is unclear how these demographic patterns influence muskox genetic diversity and fitness. We compared the genetic diversity of Canadian muskox populations undergoing opposing population trends. Genotyping 84 mainland and 244 Victoria Island individuals at ten microsatellite loci revealed low genetic variation (Victoria Island, mean allelic richness 1.66, expected heterozygosity 0.16; mainland, mean allelic richness 2.58, expected heterozygosity 0.41), with no evidence of further reductions in diversity subsequent to recent demographic declines. Bayesian modelling showed that a 1900s bottleneck contributed to the lack of diversity in contemporary populations, and forward-in-time simulations suggested little effect on genetic diversity over the next 100 years. Muskoxen might have reached a genetic diversity minimum, and additional research will be needed to determine their capacity to adapt to rapid changes in selective pressures in a rapidly changing Arctic.

ONRAB® is a human adenovirus rabies glycoprotein recombinant vaccine developed to control rabies in wildlife. To support licensing and widespread use of the vaccine, safety studies are needed to assess its potential residual impact on wildlife populations. We examined the persistence of the ONRAB® vaccine virus in captive rabies vector and non-target mammals. This research complements work on important rabies vector species (raccoon, striped skunk, and red fox) but also adds to previous findings with the addition of some non-target species (Virginia opossum, Norway rats, and cotton rats) and a prolonged period of post vaccination monitoring (41 days). Animals were directly inoculated orally with the vaccine and vaccine shedding was monitored using quantitative real-time PCR applied to oral and rectal swabs. ONRAB® DNA was detected in both oral and rectal swabs from 6 h to 3 days post-inoculation in most animals, followed by a resurgence of shedding between days 17 and 34 in some species. Overall, the duration over which ONRAB® DNA was detectable was shorter for non-target mammals, and by day 41, no animal had detectable DNA in either oral or rectal swabs. All target species, as well as cotton rats and laboratory-bred Norway rats, developed robust humoral immune responses as measured by competitive ELISA, with all individuals being seropositive at day 31. Similarly, opossums showed good response (89% seropositive; 8/9), whereas only one of nine wild caught Norway rats was seropositive at day 31. These results support findings of other safety studies suggesting that ONRAB® does not persist in vector and non-target mammals exposed to the vaccine. As such, we interpret these data to reflect a low risk of adverse effects to wild populations following distribution of ONRAB® to control sylvatic rabies.

Populations are exposed to different types and strains of pathogens across heterogeneous landscapes, where local interactions between host and pathogen may present reciprocal selective forces leading to correlated patterns of spatial genetic structure. Understanding these coevolutionary patterns provides insight into mechanisms of disease spread and maintenance. Arctic rabies (AR) is a lethal disease with viral variants that occupy distinct geographic distributions across North America and Europe. Red fox (Vulpes vulpes)are a highly susceptible AR host, whose range overlaps both geographically distinct AR strains and regions where AR is absent. It is unclear if genetic structure exists among red fox populations relative to the presence/absence of AR or the spatial distribution of AR variants. Acquiring these data may enhance our understanding of the role of red fox in AR maintenance/spread and inform disease control strategies. Using a genotyping-by-sequencing assay targeting 116 genomic regions of immunogenetic relevance, we screened for sequence variation among red fox populations from Alaska and an outgroup from Ontario, including areas with different AR variants, and regions where the disease was absent. Presumed neutral SNP data from the assay found negligible levels of neutral genetic structure among Alaskan populations. The immunogenetically-associated data identified 30 outlier SNPs supporting weak to moderate genetic structure between regions with and without AR in Alaska. The outliers included SNPs with the potential to cause missense mutations within several toll-like receptor genes that have been associated with AR outcome. In contrast, there was a lack of genetic structure between regions with different AR variants. Combined, we interpret these data to suggest red fox populations respond differently to the presence of AR, but not AR variants. This research increases our understanding of AR dynamics in the Arctic, where host/disease patterns are undergoing flux in a rapidly changing Arctic landscape, including the continued northward expansion of red fox into regions previously predominated by the arctic fox (Vulpes lagopus).

Frog virus 3 (FV3) and FV3-like ranaviruses can infect a variety of cold-blooded aquatic species and present a primary threat to amphibians across the globe. Previous studies of FV3-like viruses have largely investigated higher-level phylogenetic distinctions of these pathogens via portions of the conserved major capsid protein (MCP), and the putative virulence gene vIF-2α. Few studies, however, have investigated the spatial distribution of FV3 variants at the population level3-data that can be used to further understand the spatial epidemiology of this disease. In this study, we sequenced the MCP and vIF-2α of 127 FV3-positive amphibians sampled from Canadian water bodies in Ontario, northeastern Alberta, and southern Northwest Territories to explore whether intraspecific genetic variation exists within FV3. There was a lack of variation at the 2 markers across these regions, suggesting that there is a lack of FV3 sequence diversity in Canada, which may hint at a single source of infection that has spread. However, an undocumented variant termed Wood Buffalo ranavirus (WBRV) was detected in samples from 3 sites in Alberta and Northwest Territories that clustered within the FV3-like lineage with 99.3% sequence homology for MCP. For vIF-2α, all sequences were the expected truncated variant except for 6 samples in Ontario. These latter sequences were suggestive of recombination with common midwife toad virus (CMTV). The lack of variation suggests that higher-resolution genome analyses will be required to further explore the spatial spread and intraspecific variation of the disease.

Pseudogymnoascus destructans is the causal agent of bat white-nose syndrome (WNS), which is devastating some North American bat populations. Previous transcriptome studies provided insight regarding the molecular mechanisms involved in WNS; however, it is unclear how different environmental parameters could influence pathogenicity. This information could be useful in developing management strategies to mitigate the negative impacts of P. destructans on bats. We cultured three P. destructans isolates from Atlantic Canada on two growth media (potato dextrose agar and Sabouraud dextrose agar) that differ in their nitrogen source, and at two separate incubation temperatures (4 C and 15 C) that approximate the temperature range of bat hibernacula during the winter and a temperature within its optimal mycelial growth range. We conducted RNA sequencing to determine transcript levels in each sample and performed differential gene expression (DGE) analyses to test the influence of growth medium and incubation temperature on gene expression. We also compared our in vitro results with previous RNA-sequencing data sets generated from P. destructans growing on the wings of a susceptible host, Myotis lucifugus. Our findings point to a critical role for substrate and incubation temperature in influencing the P. destructans transcriptome. DGE analyses suggested that growth medium plays a larger role than temperature in determining P. destructans gene expression and that although the psychrophilic fungus responds to different nitrogen sources, it may have evolved for continued growth at a broad range of low temperatures. Further, our data suggest that down-regulation of the RNA-interference pathway and increased fatty acid metabolism are involved in the P. destructans-bat interaction. Finally, we speculate that to reduce the activation of host defense responses, P. destructans minimizes changes in the expression of genes encoding secreted proteins during bat colonization.

Road mortality is identified as a threat to American badger (Taxidea taxus (Schreber, 1777)) populations across Canada. Understanding habitat selection and movement in relation to roads is therefore vital to their conservation. Using telemetry data and road-kill locations of badgers in southwestern Ontario, we examined the relationship between habitat selection, movement patterns, and roads at three spatial scales. At the study-area scale, we assessed the effects of habitat attributes on burrow site selection. Several individuals placed their burrows closer to primary highways than expected, suggesting that badgers are not sensitive to human disturbances at this scale. Using straight-line movement trajectories between burrows, we analyzed individual movement patterns within home ranges. All badgers showed some degree of road avoidance, either crossing fewer roads or roads that posed lower mortality risk. At the road-crossing scale, we compared landscape features surrounding road-kill locations to random locations along the same roadway. There was a positive relationship between road-kill locations and number of water-based linear features and higher density of hedgerow cover. Our results provide evidence that badger movement is influenced by roads at multiple scales, which has important implications for managers interested in developing road-mitigation strategies for this endangered population.

White‐nose syndrome (WNS) has devastated populations of hibernating bats in eastern North America, leading to emergency conservation listings for several species including the previously ubiquitous little brown myotis (Myotis lucifugus). However, some bat populations near the epicenter of the WNS panzootic appear to be stabilizing after initial precipitous declines, which could reflect a selective immunogenetic sweep. To investigate the hypothesis that WNS exerts significant selection on the immunome of affected bat populations, we developed a novel, high‐throughput sequence capture assay targeting 138 adaptive, intrinsic, and innate immunity genes of putative adaptive significance, as well as their respective regulatory regions (~370 kbp of genomic sequence/individual). We used the assay to explore baseline immunogenetic variation in M. lucifugus and to investigate whether particular immune genes/variants are associated with WNS susceptibility. We also used our assay to detect 1,038 putatively neutral single nucleotide polymorphisms and characterize contemporary population structure, providing context for the identification of local immunogenetic adaptation. Sequence capture provided a cost‐effective, “all‐in‐one” assay to test for neutral genetic and immunogenetic structure and revealed fine‐scale, baseline immunogenetic differentiation between sampling sites

Ranaviruses are pathogens associated with the decline of amphibian populations across much of their distribution. In North America, frog virus 3 (FV3) is a widely distributed pathogen with wild populations of amphibians harboring different lineages and putative recombinants between FV3 and common midwife toad virus (CMTV). These recombinants have higher pathogenicity, and CMTV-derived genes associated with virulence are reported in wild strains in Canada. However, while FV3 is linked to amphibian die-offs in North America, CMTVs have been reported only in commercial frog farms in North America. We sequenced complete genomes of 18 FV3 isolates from three amphibian species to characterize genetic diversity of the lineages in Canada and infer possible recombinant regions. The 18 FV3 isolates displayed different signals of recombination, varying from none to interspersed recombination with previously isolated CMTV-like viruses. In general, most recombination breakpoints were located within open reading frames (ORFs), generating new ORFs and proteins that were a mixture between FV3 and CMTV. A combined spatial and temporal phylogeny suggests the presence of the FV3 lineage in Canada is relatively contemporary (

American badgers ( Taxidea taxus jacksoni) at the periphery of the species' range in Ontario, Canada, are listed as endangered because of an estimated population size of200 mature individuals. The main threats faced by this population include habitat loss and road mortality. However, on 18 November 2013, a radio-implanted badger was found nonresponsive in an agricultural field with signs consistent with canine distemper virus infection, which was subsequently confirmed. This prompted our investigation into the occurrence of pathogens in this endangered carnivore to better quantify the level of risk infectious disease poses to population persistence. We examined serum samples from nine live-trapped individuals and 27 whole badger specimens submitted for postmortem examination. We found evidence of exposure to canine distemper virus, canine parvovirus, and leptospires. However, infection associated with disease was not the leading cause of mortality. Future research into the effects of disease on kit survival and a comprehensive understanding of disease severity and spread from reservoir populations (e.g., raccoons [ Procyon lotor ] and striped skunks [ Mephitis mephitis ]) to badgers will be of particular importance to the conservation of this endangered population.

Blow fly (Diptera: Calliphoridae) larvae are commonly used in forensic cases to determine postmortem intervals using development rates and successional changes in community composition. Studies are conducted from different regions to provide these data. We wanted to know how widely applicable these data are. We examined whether urbanized landscapes have distinct urban blow fly communities or whether the community composition in urbanized areas is simply a variation of that found in the surrounding habitat or ecozone. Using liver baited traps, we sampled 7,272 flies from 32 sites across Canada and used mapping analysis to assess urban and rural landcover classifications, and compared urban and rural species abundance and composition. Blow fly species communities from urban areas across Canada were made up of similar species and differed from the communities found in nearby rural sites. Trapping at rural sites caught more blow flies compared with urban sites (mean flies/site 59.5 and 12.4). Of the 14 species caught, 8 were caught at urban sites, 61% of these being Cynomya cadaverina Robineau-Desvoidy, 14% Phormia regina Meigen, and 11% Lucilia sericata (Meigen). In rural sites, all 14 species were caught, 41% of specimens caught were P. regina, 21% C. cadaverina, 10% Calliphora vomitoria (Linnaeus), with only 4% L. sericata. These data suggest that regional studies are appropriate for forensic entomology applications in urban landscapes, given the similar trends across Canada, less so for wilderness or rural landscapes.

Spillover of viruses from bats to other animals may be associated with increased contact between them, as well as increased shedding of viruses by bats. Here, we tested the prediction that little brown bats (Myotis lucifugus) co-infected with the M. lucifugus coronavirus (Myl-CoV) and with Pseudogymnoascus destructans (Pd), the fungus that causes bat white-nose syndrome (WNS), exhibit different disease severity, viral shedding and molecular responses than bats infected with only Myl-CoV or only P. destructans. We took advantage of the natural persistence of Myl-CoV in bats that were experimentally inoculated with P. destructans in a previous study. Here, we show that the intestines of virus-infected bats that were also infected with fungus contained on average 60-fold more viral RNA than bats with virus alone. Increased viral RNA in the intestines correlated with the severity of fungus-related pathology. Additionally, the intestines of bats infected with fungus exhibited different expression of mitogen-activated protein kinase pathway and cytokine related transcripts, irrespective of viral presence. Levels of coronavirus antibodies were also higher in fungal-infected bats. Our results suggest that the systemic effects of WNS may down-regulate anti-viral responses in bats persistently infected with M. lucifugus coronavirus and increase the potential of virus shedding.

Protophormia terraenovae (Robineau-Desvoidy) (Diptera: Calliphoridae) and Phormia regina (Meigen) (Diptera: Calliphoridae) are morphologically similar blow fly species commonly used for estimating postmortem intervals. Field collection and storage of adults can result in color changes, in particular on calypters and palps; often collected specimens show damage such as wing fray or fungal growth. We measured the frons width: total head width ratio using photographs (ImageJ version 1.49) to differentiate these two species. Both sexes were distinguishable to species, with the greatest difference between males: 12.34% P. terraenovae versus 1.62% P. regina, less so for females: 40.25% P. terraenovae, versus 33.65% P. regina. Incorporating this feature into future blow fly keys would help with distinguishing field-caught specimens when other features are obstructed.

Humans have long relied on ungulates for food, clothing, manual labor, and transportation. Ungulates were among the first species to be domesticated and managed in the wild, but more than one-third of species are currently of conservation concern. Starting in the late twentieth century, ungulate research and management began employing genetic tools to assess attributes like the degree of population structure, inbreeding, and variation in functionally important genes. As sequencing technology advanced, research on ungulates shifted to now assay variation across the entire genome. More than 20 ungulates have had their genome assembled with a mean length of 2.6 Gb and N50 of 26 Mb. Genomic studies have provided deeper insights into the evolutionary relationships among giraffes and bovids, while camelids and horses have had their entire species demographic histories reconstructed using novel Markovian coalescent models. Moreover, artificial and natural selection has left clear signatures on ungulate genomes with high-throughput sequencing techniques being used to identify the genetic basis to important phenotypic traits. Novel assembly strategies and genomic assays are regularly being employed on ungulates, and research on this ecological and economically valuable group will help chart the course of the emerging field of wildlife genomics.

Forensic entomology involves the use of insects and arthropods to assist a spectrum of medico-criminal investigations that range from identifying cases of abuse, corpse movements, and most commonly, post mortem interval estimates. Many of these applications focus on the use of blowflies given their predicable life history characteristics in their larval stages. Molecular tools have become increasingly important in the unambiguous identification of larval blowfly species, however, these same tools have the potential to broaden the array of molecular applications in forensic entomology to include individual identifications and population assignments. Herein, we establish a microsatellite profiling system for the blowfly, Phormia regina (Diptera: Calliphoridae). The goal being to create a system to identify the population genetic structure of this species and subsequently establish if these data are amenable to identifying corpse movements based on the geographic distribution of specific genetic clusters of blowflies. Using next generation sequencing technology, we screened a partial genomic DNA sequence library of P. regina, searching for di-, tetra-, and penta-nucleotide microsatellite loci. We identified and developed primers for 84 highly repetitive segments of DNA, of which 14 revealed consistent genotypes and reasonable levels of genetic variation (4–26 alleles/locus; heterozygosity ranged from 0.385 to 0.909). This study provides the first step in assessing the utility of microsatellite markers to track the movements and sources of corpses via blowflies.

Zoonotic wildlife diseases pose significant health risks not only to their primary vectors but also to humans and domestic animals. Rabies is a lethal encephalitis caused by rabies virus (RV). This RNA virus can infect a range of terrestrial mammals but each viral variant persists in a particular reservoir host. Active management of these host vectors is needed to minimize the negative impacts of this disease, and an understanding of the immune response to RV infection aids strategies for host vaccination. Current knowledge of immune responses to RV infection comes primarily from rodent models in which an innate immune response triggers activation of several genes and signalling pathways. It is unclear, however, how well rodent models represent the immune response of natural hosts. This study investigates the innate immune response of a primary host, the raccoon, to a peripheral challenge using the raccoon rabies virus (RRV). The extent and temporal course of this response during RRV infection was analysed using genes predicted to be upregulated during infection (IFNs; IFN regulatory factors; IL-6; Toll like receptor-3; TNF receptor). We found that RRV activated components of the innate immune system, with changes in levels of transcripts correlated with presence of viral RNA. Our results suggest that natural reservoirs of rabies may not mimic the immune response triggered in rodent models, highlighting the need for further studies of infection in primary hosts.

Rabies virus presents a global public health problem. Our current understanding of the molecular determinants of rabies virulence stems from rodent models and laboratory strains of the virus, however, it is unclear how well rodent models represent viral response in natural reservoirs. Here, we examined interactions between the raccoon variant of rabies virus (RRV) and its natural host, raccoons, to gain a better understanding of molecular determinants of virulence in this system. We found expression patterns of RRV genes under tight control until the virus reached the central nervous system where replication increased significantly. Further, our examination of viral variants within an individual revealed that variant diversity may have an effect on virulence. We found that a mutation at a region of a T helper cell epitope on the nucleoprotein was associated with viral challenge outcomes and could be associated with RRV pathogenicity.

In migratory birds, population-genetic structure is generally low, but philopatric species can have fine-scale patterns of differentiation. We investigated the population-genetic structure of the Canada Goose (Branta canadensis) across multiple spatial scales to determine whether genetic data support current delineations of subspecies and populations for management. We collected samples from two subspecies (B. c. interior and B. c.maxima) and four management populations across Ontario and Quebec. Using 7 microsatellites and 442 base pairs of mtDNA we found that genetic structure varied with scale. FST revealed low levels of genetic differentiation between subspecies and management populations, and individual-based clustering revealed no genetic differentiation. However, fine-scale spatial autocorrelation revealed significant levels of relatedness at distances

Spatially explicit capture–recapture (SECR) models are gaining popularity for estimating densities of mammalian carnivores. They use spatially explicit encounter histories of individual animals to estimate a detection probability function described by two parameters: magnitude (g 0), and spatial scale (σ). Carnivores exhibit heterogeneous detection probabilities and home range sizes, and exist at low densities, so g 0 and σ likely vary, but field surveys often yield inadequate data to detect and model the variation. We sampled American black bears (Ursus americanus) on 43 study areas in ON, Canada, 2006–2009. We detected 713 animals 1810 times; however, study area-specific samples were sometimes small (6–34 individuals detected 13–93 times). We compared AIC c values from SECR models fit to the complete data set to evaluate support for various forms of variation in g 0 and σ, and to identify a parsimonious model for aggregating data among study areas to estimate detection parameters more precisely. Models that aggregated data within broad habitat classes and years were supported over those with study area-specific g 0 and σ (ΔAIC c ≥ 30), and precision was enhanced. Several other forms of variation in g 0 and σ, including individual heterogeneity, were also supported and affected density estimates. If study design cannot eliminate detection heterogeneity, it should ensure that samples are sufficient to detect and model it. Where this is not feasible, combing sparse data across multiple surveys could allow for improved inference.

Biogeochemical markers in ecology are useful for indicating geographic origin and movement patterns of species on various temporal and spatial scales. By assessing these markers in a tissue that is chemically inert once formed (e.g., claws) and is grown incrementally (i.e., deposited in layers), changes in an individual's foraging environment would be captured in the chemical signatures in these tissues. To determine whether trace elements can be used as a marker to track movement in terrestrial carnivores, we resolved multi-elemental data at a fine spatial scale in the claw keratin of a model species (Taxidea taxus) using high resolution laser ablation inductively coupled plasma mass spectrometry (LA ICP-MS). The unique chemical signatures we detected in claws suggested that these chemical variations were, in part, a reflection of the chemical environment and were not be attributable to sex or random deposition. Neighbouring claw samples were not more chemically related than distant ones, which suggests ...

[Aim] Understanding colonization dynamics is crucial for management of invasive species. We compare the genetic structure of historical (Central Europe) and recent (Spain) invasive populations with native and captive populations of the North American raccoon (Procyon lotor). Our aim was to analyse the effects of colonization age on genetic population structure, understand the role of captive individuals as potential founders and test the role of rivers for the dispersal of the species., [Location] North America, Spain, Central Europe., [Methods] We genotyped wild‐caught raccoons from Spain and Central Europe (N = 596), zoos (N = 57) and the native range (N = 153) at 16 microsatellite loci and sequenced a mitochondrial DNA fragment (Control Region). We analysed population genetic structuring with Bayesian assignment methods and a FCA. In a landscape genetic analysis, we tested the effect of waterways in the dispersal of the species., [Results] We detected 16 genetic clusters (in baps), supporting the hypothesis of multiple introductions and ongoing releases in the invasive range. The native population showed nearly no genetic structure, the Central European clusters showed signals of admixture, whereas the Spanish clusters were clearly separated. Admixture of the Central European clusters was probably caused by recent contact of populations with different origin. The landscape genetic analysis showed that rivers represent neither barriers nor corridors in Central Europe., [Main conclusions] As the Spanish populations are genetically more diverse than the Central European, we expect increased within‐population diversity when the still isolated populations merge after range expansion. As our results provide evidence for gene flow between zoos and free‐ranging populations, better control of pet trade is essential in the management efforts concerning this invasive species. Our study shows that genetic analyses can help to reconstruct invasion processes, which is important for better understanding and effective management of invasive species., The work was funded by the German Science Foundation (DFG, GRK 1319) as part of the interdisciplinary graduate school ‘Cooperation of Science and Jurisprudence in Improving Development and Use of Standards for Environmental Protection – Strategies for Risk Assessment and Management’.

The processes leading to genetic isolation influence a population’s local extinction risk, and should thus be identified before conservation actions are implemented. Natural or human-induced circumstances can result in historical or contemporary barriers to gene flow and/or demographic bottlenecks. Distinguishing between these hypotheses can be achieved by comparing genetic diversity and differentiation in isolated vs. continuous neighboring populations. In Ontario, American black bears (Ursus americanus) are continuously distributed, genetically diverse, and exhibit an isolation-by-distance structuring pattern, except on the Bruce Peninsula (BP). To identify the processes that led to the genetic isolation of BP black bears, we modelled various levels of historical and contemporary migration and population size reductions using forward simulations. We compared simulation results with empirical genetic indices from Ontario black bear populations under different levels of geographic isolation, and conducted additional simulations to determine if translocations could help achieve genetic restoration. From a genetic standpoint, conservation concerns for BP black bears are warranted because our results show that: i) a recent demographic bottleneck associated with recently reduced migration best explains the low genetic diversity on the BP; and ii) under sustained isolation, BP black bears could lose between 70% and 80% of their rare alleles within 100 years. Although restoring migration corridors would be the most effective method to enhance long-term genetic diversity and prevent inbreeding, it is unrealistic to expect connectivity to be re-established. Current levels of genetic diversity could be maintained by successfully translocating 10 bears onto the peninsula every 5 years. Such regular translocations may be more practical than landscape restoration, because areas connecting the peninsula to nearby mainland black bear populations have been irreversibly modified by humans, and form strong barriers to movement.

Small and isolated populations often exhibit low genetic diversity due to drift and inbreeding, but may simultaneously harbour adaptive variation. We investigate spatial distributions of immunogenetic variation in American badger subspecies (Taxidea taxus), as a proxy for evaluating their evolutionary potential across the northern extent of the species’ range. We compared genetic structure of 20 microsatellites and the major histocompatibility complex (MHC DRB exon 2) to evaluate whether small, isolated populations show low adaptive polymorphism relative to large and well‐connected populations. Our results suggest that gene flow plays a prominent role in shaping MHC polymorphism across large spatial scales, while the interplay between gene flow and selection was stronger towards the northern peripheries. The similarity of MHC alleles within subspecies relative to their neutral genetic differentiation suggests that adaptive divergence among subspecies can be maintained despite ongoing gene flow along subspecies boundaries. Neutral genetic diversity was low in small relative to large populations, but MHC diversity within individuals was high in small populations. Despite reduced neutral genetic variation, small and isolated populations harbour functional variation that likely contribute to the species evolutionary potential at the northern range. Our findings suggest that conservation approaches should focus on managing adaptive variation across the species range rather than protecting subspecies per se.

Characterization and population genetic analysis of multilocus genes, such as those found in the major histocompatibility complex (MHC) is challenging in nonmodel vertebrates. The traditional method of extensive cloning and Sanger sequencing is costly and time-intensive and indirect methods of assessment often underestimate total variation. Here, we explored the suitability of 454 pyrosequencing for characterizing multilocus genes for use in population genetic studies. We compared two sample tagging protocols and two bioinformatic procedures for 454 sequencing through characterization of a 185-bp fragment of MHC DRB exon 2 in wolverines (Gulo gulo) and further compared the results with those from cloning and Sanger sequencing. We found 10 putative DRB alleles in the 88 individuals screened with between two and four alleles per individual, suggesting amplification of a duplicated DRB gene. In addition to the putative alleles, all individuals possessed an easily identifiable pseudogene. In our system, sequence variants with a frequency below 6% in an individual sample were usually artefacts. However, we found that sample preparation and data processing procedures can greatly affect variant frequencies in addition to the complexity of the multilocus system. Therefore, we recommend determining a per-amplicon-variant frequency threshold for each unique system. The extremely deep coverage obtained in our study (approximately 5000×) coupled with the semi-quantitative nature of pyrosequencing enabled us to assign all putative alleles to the two DRB loci, which is generally not possible using traditional methods. Our method of obtaining locus-specific MHC genotypes will enhance population genetic analyses and studies on disease susceptibility in nonmodel wildlife species.

Conservation strategies have a tendency to discount range peripheries, but recent evidence suggests that range edges may be important to species persistence by harboring genetic variants not found in core distributions. Wolverines in Canada are recognized as existing in two populations—an endangered eastern population and an extant western population thought to be largely panmictic. Studies from western North America identified strong patterns of female philopatry and increased genetic structure at the current southwestern periphery. Due to the paucity of data from the contemporary eastern periphery, it remains unclear if similar patterns exist at this range edge. Using neutral microsatellite and mitochondrial DNA markers from a broad geographic extent (>2,500 km), we found that wolverines at the eastern periphery displayed strong patterns of genetic distinctiveness from northwestern populations. While the microsatellite data suggest contemporary genetic structure exists, the haplotypic composition of the eastern periphery drastically differed from the core, indicating longstanding differences between regions. Further research is needed to determine if wolverines from the eastern periphery show evidence of interactions with the functionally extirpated eastern population, if functional markers display similar patterns of genetic diversity, and what relevance these may have in their evolutionary potential. Pronounced environmental fluctuations at range boundaries likely contribute to peripheral populations having genotypes with a greater capacity to respond to future selection pressures like climate change and may become a vital source of genetic diversity should core regions become replaced by edge habitats, and thus warrant separate conservation consideration.

In the absence of obvious barriers to dispersal microsatellite studies of vagile mammalian carnivores frequently find panmictic-like genetic structure over wide scales, whereas high levels of differentiation at much finer scales are detected with mitochondrial DNA (mtDNA). Given the maternal inheritance of mtDNA, these differences are often attributed to male-biased dispersal, remnants of postglacial range expansion, or both. Based on such contrasting results, it is not always clear how to delineate contemporary populations. We investigated the genetic structure of American black bears (Ursus americanus) over a wide geographic area (.1,700 km) that has no obvious physiogeographic barriers to gene flow. We analyzed a 315-base pair fragment of the mtDNA control region from 660 individual bears from 23 regions of Ontario, Canada. Relative to black bear studies based on nuclear data, mitochondrial analyses revealed much stronger patterns of genetic structure among regions (0.09 , FST , 0.44), even at small-scale intervals (,150 km), which likely reflects strong female philopatry combined with male-biased dispersal. The patterns of genetic differentiation among regions were consistent with previously described historical patterns in black bears, specifically the division of the species into 2 phylogeographic clades (coastal and continental). We confirmed that further subdivision of the continental clade occurs in a region where obvious physiogeographic barriers do not exist. We postulate that this small-scale differentiation can be explained by residual patterns from postglacial recolonization routes on either side of the Great Lakes. We suggest that it was maintained through extreme female philopatry due to habitat saturation following the postglacial geographic expansion. Based on our results, we propose that a combination of several molecular markers can be more useful in defining population units for conservation and management decisions than biparentally inherited microsatellites.

Many highly mobile species, such as migratory birds, can move and disperse over long distances, yet exhibit high levels of population genetic structuring. Although movement capabilities may enable dispersal, gene flow may be restricted by behavioural constraints such as philopatry. In the present study, we examined patterns of genetic differentiation across the range of a highly mobile, colonial waterbird. American white pelicans (Pelecanus erythrorhynchos) breed across continental North America and are currently experiencing a range expansion, especially on the eastern range limit. To assess patterns of genetic structuring, we sampled 333 individuals from 19 colonies across their North American range. The use of ten variable microsatellite markers revealed high levels of allelic richness with no population differentiation. Both Bayesian and frequentist approaches to examining genetic structuring revealed a single panmictic population. We found no evidence of genetic structuring across the Continental Divide or between migratory and non-migratory colonies. The lack of any genetic structure across the range indicates that, unlike other waterbirds with similar life-history characteristics, extensive gene flow and presumably low philopatry appear to preclude genetic differentiation. The lack of population genetic structure in American white pelicans provides an example of range-wide panmixia, a rare phenomenon in any terrestrial species. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 102, 583‐592.

We examined the mitochondrial genetic structure of American white pelicans (Pelecanus erythrorhynchos) to: 1) verify or refute whether American white pelicans are panmictic and 2) understand if any lack of genetic structure is the result of contemporary processes or historical phenomena. Sequence analysis of mitochondrial DNA control region haplotypes of 367 individuals from 19 colonies located across their North American range revealed a lack of population genetic or phylogeographic structure. This lack of structure was unexpected because: 1) Major geographic barriers such as the North American Continental Divide are thought to limit dispersal; 2) Differences in migratory behavior are expected to promote population differentiation; and 3) Many widespread North American migratory bird species show historic patterns of differentiation resulting from having inhabited multiple glacial refugia. Further, high haplotype diversity and many rare haplotypes are maintained across the species' distribution, despite frequent local extinctions and recolonizations that are expected to decrease diversity. Our findings suggest that American white pelicans have a high effective population size and low natal philopatry. We suggest that the rangewide panmixia we observed in American white pelicans is due to high historical and contemporary gene flow, enabled by high mobility and a lack of effective physical or behavioral barriers.

Several American badger subspecies are of conservation concern and most notably Taxidea taxus jacksoni and T. t. jeffersonii are both considered endangered in Canada. Previous studies report extremely low genetic diversity in T. t. jacksoni making individual genetic identifications from non-invasive samples challenging with existing markers. We used 454 next generation sequencing to identify additional panels of polymorphic microsatellites for the American badger. We designed nineteen dinucleotide microsatellite primers of >17 uninterrupted repeat motifs. Of these markers, only eight showed polymorphism in T. t. jacksoni. We also amplified these loci in T. t. jeffersonii and T. t. taxus. Total allele number per locus ranged from 2 to 11 and expected heterozygosity from 0.3 to 0.87. T. t. taxus showed higher heterozygosity across loci, whereas T. t. jacksoni showed the lowest genetic diversity. These microsatellites will provide an enhanced capacity for identifying closely related individuals from non-invasive samples.

We review the role of biogeochemical signatures, such as stable isotopes and trace elements, in the cornified claw tissue as a means of studying movement and foraging behaviour of vertebrates because this approach is noninvasive and can capture contemporary and historic signatures. Because biogeochemical techniques are still relatively new in studies of animal movement, we are only beginning to understand how the growth patterns of the cornified claw sheath may affect our ability to interpret the biogeochemical signals in these tissues. To move towards resolving this, we review the morphology of the epidermal cornified claw sheath in several taxa that illustrate substantial variation in growth patterns both between taxa and between individual distinct claw regions. For instance, in mammalian claws, deposition of keratinizing cells from the epidermis is nonlinear because the claw tip is composed of old and new cornified epidermal cells, whereas the cornified blade horn covering the claw’s lateral walls is deposited continuously and without assortment, providing unbroken time-series data. We also consider patterns of growth in mammalian hooves, as well as reptilian, avian, and amphibian cornified claw sheaths, and address the need for expanded research in this field. We conclude this synthesis by describing a noninvasive technique for monitoring growth rates in a model mammal, the American badger ( Taxidea taxus (Schreber, 1777)), and provide guidelines for future sampling of claw keratin, which will improve our ability to back-calculate the time of biogeochemical integration into this tissue.

Summary 1. Population density is a critical ecological parameter informing effective wildlife management and conservation decisions. Density is often estimated by dividing capture–recapture (C–R) estimates of abundance ( ^ N) by size of the study area, but this relies on the assumption of geographic closure – a situation rarely achieved in studies of large carnivores. For geographically open populations ^ N is overestimated relative to the size of the study area because animals with only part of their home range on the study area are available for capture. This bias (‘edge effect’) is more severe when animals such as large carnivores range widely. To compensate for edge effect, a boundary strip around the trap array is commonly included when estimating the effective trap area ( ^ A). Various methods for estimating the width of the boundary strip are proposed, but ^ N ⁄ ^ A estimates of large carnivore density are generally mistrusted unless concurrent telemetry data are available to define ^ A. Remote sampling by cameras or hair snags may reduce study costs and duration, yet without telemetry data inflated density estimates remain problematic. 2. We evaluated recently developed spatially explicit capture–recapture (SECR) models using data from a common large carnivore, the American black bear Ursus americanus, obtained by remote sampling of 11 geographically open populations. These models permit direct estimation of population density from C–R data without assuming geographic closure. We compared estimates derived using this approach to those derived using conventional approaches that estimate density as ^ N ⁄ ^ A. 3. Spatially explicit C–R estimates were 20–200% lower than densities estimated as ^ N ⁄ ^ A .A ICc supported individual heterogeneity in capture probabilities and home range sizes. Variable home range size could not be accounted for when estimating density as ^ N ⁄ ^ A. 4. Synthesis and applications. We conclude that the higher densities estimated as ^ N ⁄ ^ A compared to estimates from SECR models are consistent with positive bias due to edge effects in the former. Inflated density estimates could lead to management decisions placing threatened or endangered large carnivores at greater risk. Such decisions could be avoided by estimating density by SECR when bias due to geographic closure violation cannot be minimized by study design.

A landscape genetic simulation modelling approach is used to understand factors affecting raccoon rabies disease spread in southern Ontario, Canada. Using the Ontario Rabies Model, we test the hypothesis that landscape configuration (shape of available habitat) affects dispersal, as indicated by genetic structuring. We simulated range expansions of raccoons from New York into vacant landscapes in Ontario, in two areas that differed by the presence or absence of a landscape constriction. Our results provide theoretical evidence that landscape constriction acts as a vicariant bottleneck. We discuss implications for raccoon rabies spread.