Whole‐genome resequencing uncovers molecular signatures of natural and sexual selection in wild bighorn sheep

The identification of genes influencing fitness is central to our understanding of the genetic basis of adaptation and how it shapes phenotypic variation in wild populations. Here, we used whole‐genome resequencing of wild Rocky Mountain bighorn sheep (Ovis canadensis) to >50‐fold coverage to identify 2.8 million single nucleotide polymorphisms (SNPs) and genomic regions bearing signatures of directional selection (i.e. selective sweeps). A comparison of SNP diversity between the X chromosome and the autosomes indicated that bighorn males had a dramatically reduced long‐term effective population size compared to females. This probably reflects a long history of intense sexual selection mediated by male–male competition for mates. Selective sweep scans based on heterozygosity and nucleotide diversity revealed evidence for a selective sweep shared across multiple populations at RXFP2, a gene that strongly affects horn size in domestic ungulates. The massive horns carried by bighorn rams appear to have evolved in part via strong positive selection at RXFP2. We identified evidence for selection within individual populations at genes affecting early body growth and cellular response to hypoxia; however, these must be interpreted more cautiously as genetic drift is strong within local populations and may have caused false positives. These results represent a rare example of strong genomic signatures of selection identified at genes with known function in wild populations of a nonmodel species. Our results also showcase the value of reference genome assemblies from agricultural or model species for studies of the genomic basis of adaptation in closely related wild taxa.     

Genetic Diversity and Divergence among Bighorn Sheep from Reintroduced Herds in Washington and Idaho

Bighorn sheep (Ovis canadensis) populations in the western United States have undergone widespread declines and extirpations since the late nineteenth century as a consequence of introduced diseases, competition with livestock, and unregulated hunting. Washington, Idaho, USA, and British Columbia, Canada were historically thought to be occupied by 2 bighorn lineages or subspecies: Rocky Mountain (O. c. canadensis) and California (O. c. californiana). The putative California lineage was completely extirpated in the United States, and reintroductions to reestablish populations were sourced directly or indirectly from a single region in southern British Columbia. Restoration efforts have attempted to maintain the diversity and divergence of these 2 lineages, sometimes referred to as subspecies although taxonomic classifications have changed over time. In this study we describe genetic variation in a subset of native and reintroduced herds of California and Rocky Mountain bighorn sheep. We examined genetic diversity and divergence between bighorn sheep herds using 15 microsatellite loci, including 4 loci linked to genes involved in immune function. We analyzed 504 samples from reintroduced herds in Washington (n = 10 California herds, n = 4 Rocky Mountain herds) and Idaho (n = 5 California), and source herds in Oregon (n = 1 Rocky Mountain) and British Columbia (n = 5 California, 1 Rocky Mountain). Genetic structure reflected known reintroduction history, and geographic proximity also was associated with decreased genetic divergence. Herds in Washington and Idaho sourced from California bighorn sheep were less genetically diverse than those sourced from Rocky Mountain herds. Also, levels of relatedness within and across California herds were higher than in Rocky Mountain herds and similar to what would be expected for full and half siblings. Lower diversity and higher relatedness among California herds is a concern for long-term fitness and likely related to past population bottlenecks, fewer source populations, and management history, such as entirely sourcing California herds from British Columbia. Genetic divergence of neutral loci between California and Rocky Mountain herds was greater than that of adaptive loci, potentially indicating that balancing selection has maintained similar genetic diversity across lineages in loci associated with immune and other adaptive functions. Thus, we recommend future reintroductions and augmentations should continue to use source populations from the appropriate California or Rocky Mountain lineage to avoid potential outbreeding depression and maintain possible adaptive differences. This could be accomplished by obtaining sheep from ≥1 source within the genetic lineage, while avoiding sourcing from admixed herds. Future work encompassing a broader geographic sampling of populations and a greater portion of the genome is necessary to better evaluate the degree to which contemporary divergence between lineages is associated with recent founder effects and genetic isolation or evolutionary adaptation. © 2021 The Wildlife Society     

Evaluating wildlife translocations using genomics: A bighorn sheep case study

Wildlife restoration often involves translocation efforts to reintroduce species and supplement small, fragmented populations. We examined the genomic consequences of bighorn sheep (Ovis canadensis) translocations and population isolation to enhance understanding of evolutionary processes that affect population genetics and inform future restoration strategies. We conducted a population genomic analysis of 511 bighorn sheep from 17 areas, including native and reintroduced populations that received 0–10 translocations. Using the Illumina High Density Ovine array, we generated datasets of 6,155 to 33,289 single nucleotide polymorphisms and completed clustering, population tree, and kinship analyses. Our analyses determined that natural gene flow did not occur between most populations, including two pairs of native herds that had past connectivity. We synthesized genomic evidence across analyses to evaluate 24 different translocation events and detected eight successful reintroductions (i.e., lack of signal for recolonization from nearby populations) and five successful augmentations (i.e., reproductive success of translocated individuals) based on genetic similarity with the source populations. A single native population founded six of the reintroduced herds, suggesting that environmental conditions did not need to match for populations to persist following reintroduction. Augmentations consisting of 18–57 animals including males and females succeeded, whereas augmentations of two males did not result in a detectable genetic signature. Our results provide insight on genomic distinctiveness of native and reintroduced herds, information on the relative success of reintroduction and augmentation efforts and their associated attributes, and guidance to enhance genetic contribution of augmentations and reintroductions to aid in bighorn sheep restoration.     

Temporal dynamics of linkage disequilibrium in two populations of bighorn sheep

Linkage disequilibrium (LD) is the nonrandom association of alleles at two markers. Patterns of LD have biological implications as well as practical ones when designing association studies or conservation programs aimed at identifying the genetic basis of fitness differences within and among populations. However, the temporal dynamics of LD in wild populations has received little empirical attention. In this study, we examined the overall extent of LD, the effect of sample size on the accuracy and precision of LD estimates, and the temporal dynamics of LD in two populations of bighorn sheep (Ovis canadensis) with different demographic histories. Using over 200 microsatellite loci, we assessed two metrics of multi‐allelic LD, D′, and χ^′2. We found that both populations exhibited high levels of LD, although the extent was much shorter in a native population than one that was founded via translocation, experienced a prolonged bottleneck post founding, followed by recent admixture. In addition, we observed significant variation in LD in relation to the sample size used, with small sample sizes leading to depressed estimates of the extent of LD but inflated estimates of background levels of LD. In contrast, there was not much variation in LD among yearly cross‐sections within either population once sample size was accounted for. Lack of pronounced interannual variability suggests that researchers may not have to worry about interannual variation when estimating LD in a population and can instead focus on obtaining the largest sample size possible.     

Restoration Potential of Bighorn Sheep in a Prairie Region

Efforts to recover Rocky Mountain bighorn sheep (Ovis canadensis canadensis) throughout western North America have had limited success with the majority of current populations remaining in small and isolated areas on a fraction of their historical range. Prairie environments with rugged topography throughout the Northern Great Plains ecoregion were historically occupied by relatively robust bighorn sheep populations. We predicted there is likely unrealized potential habitat for restoring bighorn sheep to these areas; however, relatively little attention has been devoted to identifying habitat in unoccupied prairie regions. We used global positioning system (GPS)-collar data collected from 43 female bighorn sheep in 2 populations located in the eastern Montana, USA, portion of the Northern Great Plains during 2014–2018 to estimate a population-level annual resource selection model and identify the important factors affecting bighorn sheep resource selection. We extrapolated model predictions across eastern Montana's prairie region and identified potential habitat to understand restoration potential and assist with future translocations of bighorn sheep. Resource selection of bighorn sheep was most strongly associated with terrain slope and ruggedness, tree canopy cover, and a normalized difference vegetation index metric. Within currently unoccupied areas of the historical range, the model extrapolation predicted 7,211 km² of habitat, with most owned and managed by private landowners (44%), Bureau of Land Management (33%), and the United States Fish and Wildlife Service (15%). Our results provide an empirical evaluation of landscape covariates influencing resource selection of bighorn sheep occupying prairie environments and provide a habitat model that may be generalizable to other areas in the Northern Great Plains ecoregion. We demonstrate substantial potential for restoration opportunities of bighorn sheep in the Northern Great Plains ecoregion. Broad restoration of bighorn sheep across the ecoregion would likely require strong collaboration among and between public resource managers, private landowners, and livestock producers given the heterogeneous land ownership patterns, management strategies, and domestic sheep distributions. © 2020 The Wildlife Society.     

Brucellosis in captive Rocky Mountain bighorn sheep (Ovis canadensis) caused by Brucella abortus biovar 4

Nine (four female, five male) captive adult Rocky Mountain bighorn sheep (Ovis canadensis) contracted brucellosis caused by Brucella abortus biovar 4 as a result of natural exposure to an aborted elk (Cervus elaphus) fetus. Clinical signs of infection were orchitis and epididymitis in males and lymphadenitis and placentitis with abortion in females. Gross pathologic findings included enlargement of the testes or epididymides, or both, and yellow caseous abscesses and pyogranulomas of the same. Brucella abortus biovar 4 was cultured in all bighorn sheep from a variety of tissues, including testes/epididymides, mammary gland, and lymph nodes. All bighorn sheep tested were positive on a variety of standard Brucella serologic tests. This is the first report of brucellosis caused by B. abortus in Rocky Mountain bighorn sheep. It also provides evidence that bighorn sheep develop many of the manifestations ascribed to this disease and that infection can occur from natural exposure to an aborted fetus from another species. Wildlife managers responsible for bighorn sheep populations sympatric with Brucella-infected elk or bison (Bison bison) should be cognizant of the possibility of this disease in bighorn sheep.     

Genomic association with pathogen carriage in bighorn sheep (Ovis canadensis)

Genetic composition can influence host susceptibility to, and transmission of, pathogens, with potential population‐level consequences. In bighorn sheep (Ovis canadensis), pneumonia epidemics caused by Mycoplasma ovipneumoniae have been associated with severe population declines and limited recovery across North America. Adult survivors either clear the infection or act as carriers that continually shed M. ovipneumoniae and expose their susceptible offspring, resulting in high rates of lamb mortality for years following the outbreak event. Here, we investigated the influence of genomic composition on persistent carriage of M. ovipneumoniae in a well‐studied bighorn sheep herd in the Wallowa Mountains of Oregon, USA. Using 10,605 SNPs generated using RADseq technology for 25 female bighorn sheep, we assessed genomic diversity metrics and employed family‐based genome‐wide association methodologies to understand variant association and genetic architecture underlying chronic carriage. We observed no differences among genome‐wide diversity metrics (heterozygosity and allelic richness) between groups. However, we identified two variant loci of interest and seven associated candidate genes, which may influence carriage status. Further, we found that the SNP panel explained ~55% of the phenotypic variance (SNP‐based heritability) for M. ovipneumoniae carriage, though there was considerable uncertainty in these estimates. While small sample sizes limit conclusions drawn here, our study represents one of the first to assess the genomic factors influencing chronic carriage of a pathogen in a wild population and lays a foundation for understanding genomic influence on pathogen persistence in bighorn sheep and other wildlife populations. Future research should incorporate additional individuals as well as distinct herds to further explore the genomic basis of chronic carriage.     

Toxoplasmic encephalitis in a free-ranging Rocky Mountain bighorn sheep from Washington

A 4-mo-old free-ranging Rocky Mountain bighorn sheep (Ovis canadensis canadensis) from the Hells Canyon area (Washington, USA) was diagnosed with encephalitis associated with Toxoplasma gondii infection. The sheep had concurrent pneumonic pasteurellosis and resided in a geographic area with endemic Pasteurella-associated pneumonia and mortality in bighorn sheep. The brain had multifocal necrotizing and nonsuppurative encephalitis with intralesional protozoa. The protozoa were identified as T. gondii by immunohistochemistry. To our knowledge, this is the first report of T. gondii infection in a Rocky Mountain bighorn sheep.     

Muellerius capillaris (Mueller, 1889) (Nematoda: Protostrongylidae): an unusual finding in Rocky Mountain bighorn sheep (Ovis canadensis canadensis Shaw) in South Dakota

Lungs and fecal samples from nine hunter-killed Rocky Mountain bighorn sheep were examined for lungworms. All samples contained adults and/or larvae of Muellerius capillaris (Mueller, 1889). Protostrongylus spp., the lungworms commonly reported from bighorn sheep, were not present in any samples. Larvae of M. capillaris bear a spine on the dorsal side of the posterior end and are shorter than dorsal-spined larvae of other lungworms recorded from North American ungulates. Larvae similar in shape but longer than those of Muellerius were found in free-ranging bighorn sheep in Alberta and British Columbia. In addition, dorsal-spined larvae have been found in bighorn sheep in Montana, North Dakota, and Washington. The identity of the dorsal-spined larvae is known only from sheep in South Dakota. Thus, caution must be taken when diagnosing lungworm infections in Rocky Mountain bighorn sheep.     

Effects of helicopter net-gunning on survival of bighorn sheep

Wildlife capture, and the data collection associated with it, has led to major advancements in ecology that are integral to decision making pertaining to wildlife conservation. Capturing wildlife, however, can cause lethal and non-lethal risks to animals. Understanding the factors that contribute to the level of risk involved in wildlife capture is therefore important for the development and implementation of the safest and most effective methodologies. We used data from 736 animal captures of 389 individuals for 2 subspecies of female bighorn sheep (Rocky Mountain bighorn [Ovis canadensis canadensis], Sierra Nevada bighorn sheep [O. c. sierrae]) in Wyoming and California, USA, in 2002–2020 to evaluate the degree and extent of time that capture via helicopter net-gunning affects survival. We compared pre- and post-capture survival during a 10-week window centered on a capture event, and post-capture survival between captured animals and animals that were monitored but not captured during the 10-week window. Additionally, we evaluated the effects of handling techniques (number of times captured, season of capture event, handling time, chase time, and body temp) and biological factors (age and nutritional condition) on probability of capture mortality. Mean daily survival was 0.9992 during a 5-week pre-capture window, dropped to 0.9864 on the day of capture, and rebounded within 3 days of capture to pre-capture levels and that of sheep that were not captured. Overall, direct mortality resulting from capture was 1.36%, with 0.54% mortality occurring within the 3 days following a capture event for an overall 1.90% capture-related mortality. The only handling and biological metrics that influenced the probability of capture mortality were rectal temperature and nutritional condition; high initial rectal temperatures and poor body condition were associated with increased risk of mortality in the days following capture. Overall, helicopter net-gunning imposed low and short-term risk to survival of female bighorn sheep. To reduce bias in survival estimates, we recommend using a 3-day censorship window for post-capture mortalities as opposed to the common practice of a 2–5-week censor window. Helicopter net-gunning, including annual or seasonal recaptures, remains an effective and comparatively safe technique for capture and associated data collection of bighorn sheep.     

Is Habitat Constraining Bighorn Sheep Restoration? A Case Study

Rocky Mountain bighorn sheep (Ovis canadensis) restoration continues to be a challenge throughout western North America despite nearly a century of efforts dedicated to the species' recovery. Though bighorn sheep restoration may be constrained by several environmental factors and behavioral tendencies, areas with unrealized restoration potential may exist if novel restoration strategies are considered. We used global positioning system (GPS) location data from 27 female bighorn sheep within the southern portion of the Madison Range in southwest Montana, USA, 2015–2017, to develop and validate winter and summer habitat models, which we extrapolated throughout the entire Madison Range to identify potential seasonal habitat. We estimated potential bighorn sheep minimum population estimates within the extrapolation area by linking our top-ranked winter habitat model to population count data. During summer, female bighorn sheep selected areas characterized by rugged and steep terrain, reduced canopy cover, southwestern aspects, and ridgelines. During winter, female bighorn sheep selected areas characterized by low elevations, southwestern aspects, steep slopes, reduced canopy cover, ridgelines, high normalized difference vegetation index amplitude, and areas close to steep terrain. Predicted summer habitat was concentrated along the high-elevation ridgelines associated with steep slopes and reduced canopy cover. Predicted winter habitat occurred in a non-contiguous distribution primarily along the low-elevation, southwest-facing aspects along the western slopes of the Madison Range. Our results suggest that the Madison Range may be capable of supporting 780–1,730 animals, which is 2–4 times the number of bighorn sheep currently observed within the range. Further, our findings provide managers with a quantification of female bighorn sheep habitat and suggests that a strategy focused on establishing a metapopulation through a series of within-range translocations may enhance bighorn sheep restoration. We suggest that similar restoration opportunities may be common in other unoccupied areas of bighorn sheep historical range. © 2020 The Wildlife Society.     

Microsatellite evolution in congeneric mammals: domestic and bighorn sheep

We compared genotypes at eight (AC)n microsatellite loci in domestic sheep (Ovis aries) and wild Rocky Mountain bighorn sheep (O. canadensis). The domestic sheep had greater genetic variation, higher allele-size variances, and larger allele sizes than the wild sheep. Accumulating evidence from higher taxonomic comparisons shows that these parameters are biased if microsatellite loci are selected in one taxon and used in another. Our results demonstrate similar biases between congeneric species. We compared standard measures of genetic variation, differentiation, and distance within and between species (H, D, FST) to newer measures based on allele-size variance (SW, SB, RST). The size-based distances better detected species-level divergence, but standard measures better distinguished allopatric populations. Empirical calibration of these measures at the subspecies level is needed to establish their useful ranges.      

Comparison of pulmonary defense mechanisms in Rocky Mountain bighorn (Ovis canadensis canadensis) and domestic sheep

Alveolar macrophages were obtained from Rocky Mountain bighorn sheep (Ovis canadensis canadensis) and domestic sheep for the purpose of comparing pulmonary host defense mechanisms in the two species. Specific variables studied included (1) characterization of the cell types present in the lung, (2) alveolar macrophage phagocytic and bactericidal functions, (3) measurement of protein levels in lavage fluid, and (4) measurement of cortisol levels in lavage fluid. While phagocytic cell populations were similar between bighorn and domestic sheep, a significantly higher percentage of lymphocytes were present in bighorns than domestics (20% in bighorn versus 6% in domestic sheep). Significant differences were not observed in the phagocytic or bactericidal functions of macrophages between the two species. Significant differences were not observed in either lavage fluid protein levels or in cortisol levels.     

Hemorrhagic disease in bighorn sheep in Arizona

Two bighorn sheep from Arizona (USA) were submitted for necropsy. One was a Rocky Mountain bighorn (Ovis canadensis canadensis) and the other was a desert bighorn (Ovis canadensis mexicana). Both had lesions consistent with those of hemorrhagic disease (HD). Epizootic hemorrhagic disease virus (EHDV) type-2 and bluetongue virus (BTV) type-17, respectively, were isolated from the sheep tissues. To our knowledge, HD caused by either EHDV or BTV infection has not been documented previously in Arizona bighorn sheep.     

Bighorn Sheep Response to Road-Related Disturbances in Rocky Mountain National Park,Colorado

ABSTRACT Bighorn sheep (Ovis canadensis) use of Sheep Lakes mineral site, Rocky Mountain National Park, Colorado, USA, has decreased since 1996. Officials were concerned that human disturbance may have been contributing to this decline in use. We evaluated effects of vehicular traffic and other road-related disturbance on bighorn use of Sheep Lakes in the summers of 2002 and 2003. We found that the time and number of attempts required by bighorn to reach Sheep Lakes was positively related to the number of vehicles and people present at Sheep Lakes. Further, the number of bighorn individuals and groups attempting to visit Sheep Lakes were negatively affected by disturbance associated with the site. The number of vehicles recorded the hour before bighorn tried to access Sheep Lakes best predicted an animal's failure to cross Fall River Road and reach Sheep Lakes. We conclude that human and road-related disturbance at Sheep Lakes negatively affected bighorn use of the mineral site. Because Sheep Lakes may be important for bighorn sheep, especially for lamb production and survival, the negative influence of disturbance may compromise health and productivity of the Mummy Range bighorn sheep.     

Susceptibility of two Rocky Mountain bighorn sheep to experimental infection with Anaplasma ovis.

In North America, the role of wild ruminants in the epidemiology of anaplasmosis has not been clearly defined. Such information is particularly meager in regard to bighorn sheep. We report the susceptibility of two Rocky Mountain bighorn sheep (Ovis canadensis canadensis) to experimental infection with a well characterized field isolate of Anaplasma ovis obtained from domestic sheep in Idaho. Both bighorn sheep developed infection resulting in severe clinical disease, with relatively high parasitemias, icterus and anemia. One animal required tetracycline therapy and responded well to treatment, while the other recovered uneventfully without treatment. Both bighorn sheep were spleen-intact, a condition that in A. ovis-exposed domestic sheep typically is associated with mild infection. The results indicate that bighorn sheep may be adversely affected if exposed to the organism in nature.     

Paratuberculosis (Johne's disease) in bighorn sheep and a Rocky Mountain goat in Colorado.

Between May, 1972 and February, 1978, six cases of paratuberculosis (Johne's Disease) caused by Mycobacterium paratuberculosis were diagnosed in free-ranging Rocky Mountain bighorn sheep (Ovis canadensis) and one Rocky Mountain goat (Oreamnos americanus) on or near Mt. Evans in Colorado. Diagnosis of paratuberculosis was based on gross and histopathologic examination of the animals and by isolation of M. paratuberculosis from three sheep and the goat. The clinical signs and pathologic changes seen in the bighorn sheep resembled those described in cattle, while the lesions in the goat were similar to those described for domestic sheep and goats.     

Desert bighorn sheep habitat selection,group size,and mountain lion predation risk

Bighorn sheep (Ovis canadensis) evolved for thousands of years in the presence of numerous predators, including mountain lions (Puma concolor). Bighorn sheep have presumably developed predator avoidance strategies; however, the effectiveness of these strategies in reducing risk of mountain lion predation is not well understood. These strategies are of increasing interest because mountain lion predation on bighorn sheep has been identified as a leading cause of mortality in some sheep populations. Therefore, we investigated how mountain lions affect both bighorn sheep habitat selection and risk of mortality in Arizona, USA. We used 2 approaches to investigate the predator-prey relationship between mountain lions and bighorn sheep. We fit 103 bighorn sheep (81 females and 22 males) with global positioning system radio-collars in 2 Arizona populations from 2013 to 2017, and used a negative binomial resource selection probability function to evaluate whether bighorn sheep selected for habitat features in accordance with presumed predator avoidance strategies, including terrain ruggedness, slope, topographic position, and horizontal obstruction, in 2 seasons (winter and summer). We then estimated how habitat features such as terrain ruggedness, slope, horizontal obstruction, and group size, influence the risk of mortality due to mountain lion predation using an Andersen-Gill proportional hazards model. Generally, both sexes selected areas with lower horizontal obstruction and intermediate ruggedness and slope, but selection patterns differed between seasons and sexes. The use of more rugged areas and steeper slopes decreased the risk of mortality due to mountain lion predation, consistent with presumed predator avoidance strategies. Increased group size decreased risk of bighorn sheep mortality due to mountain lion predation but this effect became marginal at approximately 10 individuals/group. We did not identify a relationship between horizontal obstruction and bighorn sheep mortality risk. Our findings can be used in habitat and population management decisions such as the prioritization of habitat restoration sites or selection of translocation sites. In addition, we suggest that augmentation of low-density bighorn sheep populations may reduce mountain lion predation risk by increasing group size, and that releasing large groups of bighorn sheep in population augmentation and reintroduction efforts may help to reduce mountain lion predation.     

Minimum sample sizes for population genomics: an empirical study from an Amazonian plant species

High‐throughput DNA sequencing facilitates the analysis of large portions of the genome in nonmodel organisms, ensuring high accuracy of population genetic parameters. However, empirical studies evaluating the appropriate sample size for these kinds of studies are still scarce. In this study, we use double‐digest restriction‐associated DNA sequencing (ddRADseq) to recover thousands of single nucleotide polymorphisms (SNPs) for two physically isolated populations of Amphirrhox longifolia (Violaceae), a nonmodel plant species for which no reference genome is available. We used resampling techniques to construct simulated populations with a random subset of individuals and SNPs to determine how many individuals and biallelic markers should be sampled for accurate estimates of intra‐ and interpopulation genetic diversity. We identified 3646 and 4900 polymorphic SNPs for the two populations of A. longifolia, respectively. Our simulations show that, overall, a sample size greater than eight individuals has little impact on estimates of genetic diversity within A. longifolia populations, when 1000 SNPs or higher are used. Our results also show that even at a very small sample size (i.e. two individuals), accurate estimates of F_ST can be obtained with a large number of SNPs (≥1500). These results highlight the potential of high‐throughput genomic sequencing approaches to address questions related to evolutionary biology in nonmodel organisms. Furthermore, our findings also provide insights into the optimization of sampling strategies in the era of population genomics.     

Bighorn Sheep Abundance Following a Suspected Pneumonia Epidemic in Rocky Mountain National Park

Abstract: Anecdotal evidence of a pneumonia epizootic among bighorn sheep (Ovis canadensis canadensis) in Rocky Mountain National Park (RMNP), Colorado, USA, during the mid-1990s prompted park officials to examine the current condition of the herds. Here we present a mark—resight study design to estimate population abundance that, in many circumstances, is a reliable and cost-effective alternative to traditional mark—recapture or to indices of population abundance. We captured 59 adult females and radiocollared them via helicopter net-gunning during winter 2002–2003. From ground resighting surveys conducted May—September, we estimated the total RMNP bighorn population at 389.9 (SE = 34.9, CI = 327.2–464.6) in 2003 and 366.4 (SE = 34.7, CI = 304.4–441.0) in 2004. Previous abundance estimates suggest a park-wide decline has occurred between the late 1980s and the suspected pneumonia epidemic of the mid-1990s. Although the 2 years of data from our study are not enough to predict whether the herds are capable of recovering to previous levels, they provide park officials the tools necessary to make the most informed decisions for future monitoring and management of this fragile species.