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Jennifer A. Fike

Publications and source records attributed to Jennifer A. Fike.

At least 19 recordsLinked to original sources

Designing biosurveillance using target analysis for a cryptic invasive species, the brown treesnake (Boiga irregularis)

Biosurveillance is especially important for invasive species that are difficult to control or remove once established. Target analysis, or the strategic evaluation of factors that affect or improve detection of organisms, is one approach to improve biosurveillance over time. Due to their ecologically and economically devastating effects, brown treesnakes ( Boiga irregularis ), an invasive snake in the United States territory of Guam, have a dedicated biosecurity program to prevent their spread. Guam has been occupied by brown treesnakes since the 1940s, and they are distributed throughout the island. The neighboring Saipan Island, otherwise known as Saipan, participates in biosecurity programs designed to prevent their establishment. We evaluated biosurveillance protocols within rapid response and detection exercises for brown treesnakes. Our goal was to identify, account for, and reduce potential error when inferring absence from lack of detection using data collected in Guam and Saipan. We evaluated 1) whether brown treesnake detection probability was density dependent; 2) whether using terrestrial environmental DNA ( eDNA ) sampling detected snakes in occupied areas; 3) the likelihood brown treesnakes would have been detected in Saipan, given sampling effort during a biosurveillance exercise, and whether lizards could be used as a proxy on snake-free islands for estimating factors that affect detection; 4) the effect of transect design on pacing and subsequently pacing on lizard counts; and 5) the importance of observer training in surveying for cryptic reptiles and completing broad versus targeted bioinventories. We found that brown treesnake detection probability was density dependent when using visual surveys and declined with abundance in Guam, supporting that greater effort was required to have high confidence that lack of detection supported absence in areas with low snake densities. The terrestrial eDNA sampling protocol we developed was effective at detecting snakes in occupied locations in Guam and may be a complementary tool. However, further validation of eDNA protocols would still be required for early detection application. We also found that among trained observers, the ability to detect lizards was positively predictive of the number of snakes they detected at the standard snake densities in Guam. Thus, lizard detection appeared to be a reasonable proxy for evaluating visual search protocols for snakes on snake-free islands. Overall, creating easy pace-setting relationships between time and distance (i.e., 10 m per min), with flagging that clearly marked location on a transect, greatly reduced variability in search pace across survey nights. Observers that were continuously trained to maintain brown treesnake search profiles detected twice as many, on average, cryptic reptiles compared to individuals who had never received training, which was not true for the non-reptiles (birds and small mammals). Additionally, conducting broad bioinventories with many target species reduced detection of the target species, and this was increasingly true as years since the observer completed training for the target species increased. We thus found that designing biosurveillance exercises to evaluate search protocols can help improve or refine survey methods to reduce noise in the data, and accounting for that can improve the estimated confidence that failure to detect an invasive species infers absence.

NeoBiota

Riverscape genetics of nonnative Brook Trout to inform native cutthroat trout conservation

Objective Understanding how riverscape features influence gene flow is critical for managing population connectivity in freshwater species. We examined how landscape and stream characteristics shape the spatial genetic structure of nonnative Brook Trout Salvelinus fontinalis in a headwater stream network proposed for reintroduction of federally threatened Greenback Cutthroat Trout Oncorhynchus virginalis stomias . Brook Trout were studied to evaluate the suitability of this habitat for supporting a native trout metapopulation. Methods We genotyped 757 Brook Trout from 22 sites across a 60-km stream network using 12 microsatellite loci. Spatial genetic structure was assessed using clustering analysis (program STRUCTURE) and pairwise differentiation metrics ( F ST and Jost’s D ). A spatial network modeling approach was used to quantify the effects of riverscape features (e.g., stream gradient, stream order, waterfalls, and flow direction) on trout gene flow. Results Genetic clustering identified four distinct tributary groups, while estimates of pairwise genetic differentiation indicated some genetic connectivity across the network (mean F ST = 0.04; mean Jost’s D = 0.06). Trout gene flow was impeded by waterfalls, steep stream gradients, and increased hydrologic distance. Higher stream order and downstream flow direction were associated with stronger gene flow, and stream segments containing waterfalls and steeper gradients showed greater asymmetries between upstream and downstream gene flow. Conclusions Brook Trout populations in this stream network are spatially structured, but gene flow persists and is mediated by physical riverscape features and hydrologic distance. The observed patterns of genetic connectivity suggest that this habitat can support connectivity among populations of reintroduced Greenback Cutthroat Trout. In future native trout reintroduction efforts, prioritizing habitats with gradual stream gradients and fewer waterfalls would promote population connectivity.

Transactions of the American Fisheries Society

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

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

Colorado

The potential influence of genome-wide adaptive divergence on conservation translocation outcome in an isolated greater sage-grouse population

Conservation translocations are an important conservation tool commonly employed to augment declining or reestablish extirpated populations. One goal of augmentation is to increase genetic diversity and reduce the risk of inbreeding depression (i.e., genetic rescue). However, introducing individuals from significantly diverged populations risks disrupting coadapted traits and reducing local fitness (i.e., outbreeding depression). Genetic data are increasingly more accessible for wildlife species and can provide unique insight regarding the presence and retention of introduced genetic variation from augmentation as an indicator of effectiveness and adaptive similarity as an indicator of source and recipient population suitability. We used 2 genetic data sets to evaluate augmentation of isolated populations of greater sage-grouse ( Centrocercus urophasianus ) in the northwestern region of the species range (Washington, USA) and to retrospectively evaluate adaptive divergence among source and recipient populations. We developed 2 statistical models for microsatellite data to evaluate augmentation outcomes. We used one model to predict genetic diversity after augmentation and compared these predictions with observations of genetic change. We used the second model to quantify the amount of observed reproduction attributed to transplants (proof of population integration). We also characterized genome-wide adaptive divergence among source and recipient populations. Observed genetic diversity ( H O = 0.65) was higher in the recipient population than predicted had no augmentation occurred ( H O = 0.58) but less than what was predicted by our model ( H O = 0.75). The amount of shared genetic variation between the 2 geographically isolated resident populations increased, which is evidence of periodic gene flow previously assumed to be rare. Among candidate adaptive genes associated with elevated fixation index ( F ST ) (143 genes) or local environmental variables (97 and 157 genes for each genotype–environment association method, respectively), we found clusters of genes with related functions that may influence the ability of transplants to use local resources and navigate unfamiliar environments and their reproductive potential, all possible reasons for low genetic retention from augmentation.

Conservation Biology

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

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

PLoS ONE

Genetic Connectivity in the Arizona toad (Anaxyrus microscaphus): implications for conservation of a stream dwelling amphibian in the arid Southwestern U.S.

The Arizona Toad ( Anaxyrus microscaphus ) is restricted to riverine corridors and adjacent uplands in the arid southwestern United States. As with numerous amphibians worldwide, populations are declining and face various known or suspected threats, from disease to habitat modification resulting from climate change. The Arizona Toad has been petitioned to be listed under the U.S. Endangered Species Act and was considered “warranted but precluded” citing the need for additional information – particularly regarding natural history (e.g., connectivity and dispersal ability). The objectives of this study were to characterize population structure and genetic diversity across the species’ range. We used reduced-representation genomic sequencing to genotype 3,601 single nucleotide polymorphisms in 99 Arizona Toads from ten drainages across its range. Multiple analytical methods revealed two distinct genetic groups bisected by the Colorado River; one in the northwestern portion of the range in southwestern Utah and eastern Nevada and the other in the southeastern portion of the range in central and eastern Arizona and New Mexico. We also found subtle substructure within both groups, particularly in central Arizona where toads at lower elevations were less connected than those at higher elevations. The northern and southern parts of the Arizona Toad range are not well connected genetically and could be managed as separate units. Further, these data could be used to identify source populations for assisted migration or translocations to support small or potentially declining populations.

Arizona, Nevada, New Mexico

High genetic diversity, low population genetic structure, strong natal philopatry, and longevity revealed in the Black Swift (Cypseloides niger borealis)

Genetic diversity is a critical cornerstone of biodiversity and is a central goal in management and conservation biology. Such diversity has implications for survivability, adaptability, and resiliency of a species. This study aimed to determine levels of genetic diversity and population genetic structure in the Northern Black Swift ( Cypseloides niger borealis ). This species nests across western North America as far north as Canada and south into Mexico, migrating annually to wintering areas in western Brazil. Colonies occur in isolated areas where appropriate nesting habitat is available, and in many cases, colonies are widely separated from each other. Banding data revealed that adults of this long-lived species return to their same breeding colony annually for many years. Additionally, some females exhibit natal philopatry, returning to and successfully raising offspring at the colony where they were hatched. This allowed us to hypothesize that we might expect some amount of genetic structure in the samples we studied. The life history characteristics of this species suggest that breeding colonies might be more genetically differentiated than other migratory birds that tend to have limited population genetic structure across their ranges. We used newly developed species-specific microsatellite primers to examine levels of genetic diversity and connectivity among 6 Black Swift colonies in the western United States. Levels of genetic diversity were generally high (expected heterozygosity ranging from 0.67 to 0.75) and comparable across 3 breeding sites with sufficient sample sizes ( N > 5). Principal coordinates analysis and STRUCTURE analysis showed no real clustering of individuals in regard to colonies, suggesting one panmictic metapopulation rather than multiple populations that are genetically distinct as we had previously hypothesized based on banding data. This is the first study to elucidate the genetic structure among colonies of Black Swift.

Colorado, Idaho, New Mexico

Genetic mark–recapture analysis reveals large annual variation in pre-breeding sex ratio of greater sage-grouse

Sex ratio, and the extent to which it varies over time, is an important factor in the demography, management, and conservation of wildlife populations. Greater sage-grouse Centrocercus urophasianus populations in western North America are monitored using counts of males at leks in spring. Population estimates derived from lek-count data typically assume a constant, female-biased sex ratio, yet few rigorous, empirically derived estimates of sex ratio are available to test that assumption. We estimated pre-breeding sex ratio of greater sage-grouse in a peripheral, geographically isolated population in northwestern Colorado during two consecutive winters using closed-population, robust-design, multi-state, genetic mark–recapture models in program MARK. Sex ratio varied markedly between years, with estimates of 3.29 (95% CI: 2.36–4.59) females per male in winter 2012–2013 and 1.54 (95% CI: 1.22–1.95) females per male in winter 2013–2014. Rather than assuming a constant sex ratio, biologists should consider the potential for large annual variation in sex ratio of greater sage-grouse populations when estimating population size or trend from male lek-count data.

Wildlife Biology

Assessing arthropod diversity metrics derived from stream environmental DNA: Spatiotemporal variation and paired comparisons with manual sampling

Background Benthic invertebrate (BI) surveys have been widely used to characterize freshwater environmental quality but can be challenging to implement at desired spatial scales and frequency. Environmental DNA (eDNA) allows an alternative BI survey approach, one that can potentially be implemented more rapidly and cheaply than traditional methods. Methods We evaluated eDNA analogs of BI metrics in the Potomac River watershed of the eastern United States. We first compared arthropod diversity detected with primers targeting mitochondrial 16S (mt16S) and cytochrome c oxidase 1 (cox1 or COI) loci to that detected by manual surveys conducted in parallel. We then evaluated spatial and temporal variation in arthropod diversity metrics with repeated sampling in three focal parks. We also investigated technical factors such as filter type used to capture eDNA and PCR inhibition treatment. Results Our results indicate that genus-level assessment of eDNA compositions is achievable at both loci with modest technical noise, although database gaps remain substantial at mt16S for regional taxa. While the specific taxa identified by eDNA did not strongly overlap with paired manual surveys, some metrics derived from eDNA compositions were rank-correlated with previously derived biological indices of environmental quality. Repeated sampling revealed statistical differences between high- and low-quality sites based on taxonomic diversity, functional diversity, and tolerance scores weighted by taxon proportions in transformed counts. We conclude that eDNA compositions are efficient and informative of stream condition. Further development and validation of scoring schemes analogous to commonly used biological indices should allow increased application of the approach to management needs.

PeerJ

A genetic warning system for a hierarchically structured wildlife monitoring framework

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

Ecological Applications

Simulation of genetic change under four removal strategies for a wild horse population

Small and/or artificially reduced populations can suffer genetic erosion with long-term consequences to population fitness and persistence. However, managers must periodically remove free-roaming horses from the landscape to reduce the risk of habitat degradation. We developed an individual-based population simulation model to evaluate the expected change in genetic diversity after 100 years under each of four removal strategies for the wild horse population in the Pryor Mountain management area with removals occurring every 5 years, and assuming that no additional horses are introduced to the herd. We found that long generations and high survival rates of this wild horse population guard against rapid loss of genetic diversity for all scenarios in general. However, scenarios that included a removal strategy prioritizing individuals for removal based on relatedness initially increased mean genetic diversity that was subsequently maintained at a higher level than strategies that randomly selected individuals for removal.

Report

Genetic analyses provide new insight on the mating strategies of the American Black Swift (Cypseloides niger)

Avian mating strategies play a vital role in the demographic and genetic dynamics of a species and understanding avian reproductive tactics is important to conservation, population management and restoration. Classifications of avian mating strategies have historically been based on direct physical observations and tend to be rigid population-level generalizations that overlook the variations inherent in most ecological systems. Based on limited empirical field observations, the American Black Swift Cypseloides niger borealis is considered to be a socially monogamous species with pair bonds lasting for many years. To test this hypothesis, we collected genomic DNA samples from banded swifts from six American Black Swift colonies in the western United States from 2004 to 2019 and isolated and developed primers for highly polymorphic microsatellite loci and used them to genotype our samples. Our parentage analysis revealed that sampled females never mated with the same male in subsequent years, suggesting that they are not sexually monogamous with a single partner for many years as previously hypothesized.

Colorado, Idaho, New Mexico

New strategies for characterizing genetic structure in wide-ranging, continuously distributed species: a Greater Sage-grouse case study

Characterizing genetic structure across a species’ range is relevant for management and conservation as it can be used to define population boundaries and quantify connectivity. Wide-ranging species residing in continuously distributed habitat pose substantial challenges for the characterization of genetic structure as many analytical methods used are less effective when isolation by distance is an underlying biological pattern. Here, we illustrate strategies for overcoming these challenges using a species of significant conservation concern, the Greater Sage-grouse ( Centrocercus urophasianus ), providing a new method to identify centers of genetic differentiation and combining multiple methods to help inform management and conservation strategies for this and other such species. Our objectives were to (1) describe large-scale patterns of population genetic structure and gene flow and (2) to characterize genetic subpopulation centers across the range of Greater Sage-grouse. Samples from 2,134 individuals were genotyped at 15 microsatellite loci. Using standard STRUCTURE and spatial principal components analyses, we found evidence for four or six areas of large-scale genetic differentiation and, following our novel method, 12 subpopulation centers of differentiation. Gene flow was greater, and differentiation reduced in areas of contiguous habitat (eastern Montana, most of Wyoming, much of Oregon, Nevada, and parts of Idaho). As expected, areas of fragmented habitat such as in Utah (with 6 subpopulation centers) exhibited the greatest genetic differentiation and lowest effective migration. The subpopulation centers defined here could be monitored to maintain genetic diversity and connectivity with other subpopulation centers. Many areas outside subpopulation centers are contact zones where different genetic groups converge and could be priorities for maintaining overall connectivity. Our novel method and process of leveraging multiple different analyses to find common genetic patterns provides a path forward to characterizing genetic structure in wide-ranging, continuously distributed species.

California, Colorado, Idaho, Montana, Nevada, Nort

Population genetics reveals bidirectional fish movement across the Continental Divide via an interbasin water transfer

Interbasin water transfers are becoming an increasingly common tool to satisfy municipal and agricultural water demand, but their impacts on movement and gene flow of aquatic organisms are poorly understood. The Grand Ditch is an interbasin water transfer that diverts water from tributaries of the upper Colorado River on the west side of the Continental Divide to the upper Cache la Poudre River on the east side of the Continental Divide. We used single nucleotide polymorphisms to characterize population genetic structure in cutthroat trout ( Oncorhynchus clarkii ) and determine if fish utilize the Grand Ditch as a movement corridor. Samples were collected from two sites on the west side and three sites on the east side of the Continental Divide. We identified two or three genetic clusters, and relative migration rates and spatial distributions of admixed individuals indicated that the Grand Ditch facilitated bidirectional fish movement across the Continental Divide, a major biogeographic barrier. Previous studies have demonstrated ecological impacts of interbasin water transfers, but our study is one of the first to use genetics to understand how interbasin water transfers affect connectivity between previously isolated watersheds. We also discuss implications on native trout management and balancing water demand and biodiversity conservation.

Colorado

Gene flow and spatial population structure of Brook Trout in a large headwater stream network in Colorado

We studied gene flow of non-native Brook Trout Salvelinus fontinalis in a 60-km section of continuous stream network in the upper Cache la Poudre River basin, where a large-scale reclamation effort to restore federally threatened Greenback Cutthroat Trout Oncorhynchus clarkii stomias (GBCT) is taking place. This effort—the Poudre Headwaters Project—represents the most important recovery effort of the state fish of Colorado and could result in a fivefold increase in total occupied GBCT habitat. However, the reclamation area is currently dominated by non-native Brook Trout; key steps to ensure the success of the Poudre Headwaters Project include removing non-native Brook Trout and physically isolating the area from reinvasion. We examined existing genetic population structure in the reclamation area to provide science-based guidance for effective Brook Trout removal and subsequent GBCT reintroduction. During 2018 and 2019, tissue samples were collected from 23 sites in the Long Draw region. We genotyped 792 Brook Trout using 12 microsatellite loci to examine genetic population structure within the reclamation area. Our results show that fine-scale genetic population structure is present among sites (F ST = 0.038) and overall genetic diversity is comparable to previously published estimates throughout the native range of Brook Trout (H O = 0.610; HE = 0.618). Analysis of genetic differentiation among sites indicates that sites in the reclamation area may function as a metapopulation, with fine-scale genetic structure present among tributaries that are connected by gene flow. Understanding how the landscape influences connectivity and population persistence will provide sciencebased guidance for Brook Trout removal efforts and allow managers to release GBCT in stretches of key habitat that ensure the highest probability of reintroduction success.

Colorado

Historical effective population size of North American hoary bat (Lasiurus cinereus) and challenges to estimating trends in contemporary effective breeding population size from archived samples

Background Hoary bats ( Lasiurus cinereus ) are among the bat species most commonly killed by wind turbine strikes in the midwestern United States. The impact of this mortality on species census size is not understood, due in part to the difficulty of estimating population size for this highly migratory and elusive species. Genetic effective population size (Ne) could provide an index of changing census population size if other factors affecting Ne are stable. Methods We used the NeEstimator package to derive effective breeding population size (Nb) estimates for two temporally spaced cohorts: 93 hoary bats collected in 2009–2010 and an additional 93 collected in 2017–2018. We sequenced restriction-site associated polymorphisms and generated a de novo genome assembly to guide the removal of sex-linked and multi-copy loci, as well as identify physically linked markers. Results Analysis of the reference genome with psmc suggested at least a doubling of Ne in the last 100,000 years, likely exceeding Ne = 10,000 in the Holocene. Allele and genotype frequency analyses confirmed that the two cohorts were comparable, although some samples had unusually high or low observed heterozygosities. Additionally, the older cohort had lower mean coverage and greater variability in coverage, and batch effects of sampling locality were observed that were consistent with sample degradation. We therefore excluded samples with low coverage or outlier heterozygosity, as well as loci with sequence coverage far from the mode value, from the final data set. Prior to excluding these outliers, contemporary Nb estimates were significantly higher in the more recent cohort, but this finding was driven by high values for the 2018 sample year and low values for all other years. In the reduced data set, Nb did not differ significantly between cohorts. We found base substitutions to be strongly biased toward cytosine to thymine or the complement, and further partitioning loci by substitution type had a strong effect on Nb estimates. Minor allele frequency and base quality bias thresholds also had strong effects on Nb estimates. Instability of Nb with respect to common data filtering parameters and empirically identified factors prevented robust comparison of the two cohorts. Given that confidence intervals frequently included infinity as the stringency of data filtering increased, contemporary trends in Nb of North American hoary bats may not be tractable with the linkage disequilibrium method, at least using the protocol employed here.

PeerJ

Feral horse space use and genetic characteristics from fecal DNA

Feral horses ( Equus ferus caballus ) in the western United States are managed by the Bureau of Land Management (BLM) and United States Forest Service in designated areas on public lands with a goal of maintaining populations in balance with multiple uses of the landscape. Small, isolated populations can be at risk of extirpation from stochastic events and deleterious genetic effects resulting from inbreeding and reduced heterozygosity. The genetic diversity of feral horse herds is periodically monitored using blood or hair samples collected during management gathers (i.e., occasions when the herd is rounded up). We conducted a study to examine genetic characteristics of the feral horse population at the BLM Little Book Cliffs Herd Management Area (HMA) in Colorado, USA, using non-invasively collected fecal samples. Additionally, we explored whether genotypes could be used to document space use and potential sub-population development. We used a random sampling scheme, walking transects in sampling areas covering most of the HMA to find and collect fecal samples of all ages, except those that were deteriorating. We collected >1,800 fecal samples from across the study area in May, August, and October 2014. We then identified unique individuals using a suite of microsatellite loci. Our estimates of genetic diversity from fecal samples were higher than those reported from blood and hair samples taken during recent horse gathers, likely because our sample size and spatial distribution was larger. Genotypes revealed that some individuals were found only in certain parts of the study area and at a higher proportion than random; thus, they could be considered residents in those sampling areas. Using discriminant function analyses, we detected 5 genetic groups in the sample population, but these did not correspond to individuals in specific parts of the study area. Our results support the use of fecal DNA to augment direct observations of horse presence and could be used to detect habitat use and areas of high density. Non-invasive techniques such as fecal DNA sampling can help managers decide whether new individuals need to be translocated to a closed population to maintain genetic diversity without the human safety and animal welfare concerns associated with gathers and invasive techniques.

Colorado

Composition and distribution of fish environmental DNA in an Adirondack watershed

Background Environmental DNA (eDNA) surveys are appealing options for monitoring aquatic biodiversity. While factors affecting eDNA persistence, capture and amplification have been heavily studied, watershed-scale surveys of fish communities and our confidence in such need further exploration. Methods We characterized fish eDNA compositions using rapid, low-volume filtering with replicate and control samples scaled for a single Illumina MiSeq flow cell, using the mitochondrial 12S ribosomal RNA locus for taxonomic profiling. Our goals were to determine: (1) spatiotemporal variation in eDNA abundance, (2) the filtrate needed to achieve strong sequencing libraries, (3) the taxonomic resolution of 12S ribosomal sequences in the study environment, (4) the portion of the expected fish community detectable by 12S sequencing, (5) biases in species recovery, (6) correlations between eDNA compositions and catch per unit effort (CPUE) and (7) the extent that eDNA profiles reflect major watershed features. Our bioinformatic approach included (1) estimation of sequencing error from unambiguous mappings and simulation of taxonomic assignment error under various mapping criteria; (2) binning of species based on inferred assignment error rather than by taxonomic rank; and (3) visualization of mismatch distributions to facilitate discovery of distinct haplotypes attributed to the same reference. Our approach was implemented within the St. Regis River, NY, USA, which supports tribal and recreational fisheries and has been a target of restoration activities. We used a large record of St. Regis-specific observations to validate our assignments. Results We found that 300 mL drawn through 25-mm cellulose nitrate filters yielded greater than 5 ng/µL DNA at most sites in summer, which was an approximate threshold for generating strong sequencing libraries in our hands. Using inferred sequence error rates, we binned 12S references for 110 species on a state checklist into 85 single-species bins and seven multispecies bins. Of 48 bins observed by capture survey in the St. Regis, we detected eDNA consistent with 40, with an additional four detections flagged as potential contaminants. Sixteen unobserved species detected by eDNA ranged from plausible to implausible based on distributional data, whereas six observed species had no 12S reference sequence. Summed log-ratio compositions of eDNA-detected taxa correlated with log(CPUE) (Pearson’s R = 0.655, P < 0.001). Shifts in eDNA composition of several taxa and a genotypic shift in channel catfish ( Ictalurus punctatus ) coincided with the Hogansburg Dam, NY, USA. In summary, a simple filtering apparatus operated by field crews without prior expertise gave useful summaries of eDNA composition with minimal evidence of field contamination. 12S sequencing achieved useful taxonomic resolution despite the short marker length, and data exploration with standard bioinformatic tools clarified taxonomic uncertainty and sources of error.

New York