Kinetic ELISA for detection of antibodies to Psoroptes sp. (Acari: Psoroptidae) in bighorn sheep (Ovis canadensis)

A kinetic enzyme-linked immunosorbent assay (ELISA) was developed using antigenic extracts prepared from Psoroptes cuniculi mites and sera from 37 Psoroptes sp.-infested and 43 uninfested bighorn sheep (Ovis canadensis). Serial dilutions of these serum samples, representing 3 bighorn sheep subspecies and 9 geographic areas, gave parallel responses when plotted as log dilution versus log kinetic rate. Therefore, all 80 samples were run at a single dilution (1:100) and positive/negative cutoff values were established as the mean kinetic rate of all negative sera plus either 2, 3, or 4 standard deviations. The resulting ELISA was highly reproducible and accurate with sensitivities and specificities of 100% and 97.7%, 94.6% and 97.7%, and 94.6% and 100%, respectively. This immunoassay will be useful for prospective and retrospective studies assessing the distribution and prevalence of Psoroptes sp. infestations in bighorn sheep.     

Serodiagnostic antibody responses to Psoroptes sp. infestations in bighorn sheep

The antibody responses of bighorn sheep (Ovis canadensis) infected with Psoroptes sp. mites were investigated by enzyme linked immunosorbent assay on western blots of P. cuniculi antigens. Serum from 20 Psoroptes sp.-infested bighorn sheep (O. canadensis mexicana, O. canadensis nelsoni, O. canadensis canadensis) from New Mexico, Nevada, California, and Idaho reacted strongly with mite antigens ranging from 12 to 34 kd. Serum from 35 Psoroptes sp.-free bighorn sheep of unknown tick infestation status and from three Psoroptes sp.-free bighorn sheep infested with Dermacentor hunteri ticks did not react with these antigens. Psoroptes sp.-specific antibody responses were present throughout a 16 mo period in one infected bighorn sheep, but were not detectable 8 mo following successful treatment. These results demonstrate that specific serodiagnosis of Psoroptes sp. infestation is feasible in bighorn sheep and suggest that antibody responses are indicative of current or recent infestation.     

Antigenic characterization of Psoroptes spp. (Acari: Psoroptidae) mites from different hosts

Immunoblotting with defined antigens and antisera revealed extensive and nearly complete antigenic cross-reactivity between Psoroptes spp. mites from a bighorn sheep, a mule deer, a cow, and a rabbit. Antigenic differences were not detected between mites from the sympatric bighorn sheep and mule deer. However, minor antigenic differences between mites from the cow and rabbit suggested that these mites were distinct from each other, as well as from the mites from the bighorn sheep and mule deer. These results are consistent with earlier morphologic studies of these populations of mites and provide additional support for the hypothesis that putative populations and/or species of Psoroptes mites may not be reproductively or ecologically isolated, particularly when their hosts are sympatric.     

Morphometric analysis of Psoroptes spp. mites from bighorn sheep, mule deer, cattle, and rabbits

A morphometric study of Psoroptes spp. mites was conducted to address difficulties encountered in species identification and to provide insights into the phylogenetic relationships between mites found on different hosts. A discriminant analysis employing 9 morphologic characters revealed that the lengths of the outer opisthosomal setae and the lateral margins of the opisthosomal knobs were the 2 most important characters for grouping mites according to host species. This analysis clearly separated mites collected from allopatric populations of bighorn sheep, rabbits, and cattle into discrete groups. However, differences were not detected between mites collected from sympatric populations of infested mule deer and bighorn sheep, suggesting that these mites were not host specific and represented a single interbreeding population. Differences also were not detected among mites collected from the ears and body of bighorn sheep and rabbits, demonstrating that the location of mites on a given host should not be used as a primary criterion in species identification.     

The rise and fall of psoroptic scabies in bighorn sheep in the San Andres Mountains, New Mexico

Between 1978 and 1997, a combination of psoroptic scabies (Psoroptes spp.), mountain lion (Puma concolor) predation, and periodic drought reduced a population of native desert bighorn sheep (Ovis canadensis) in the San Andres Mountains (SAM), New Mexico, from >200 individuals to a single ewe. In 1999, this ewe was captured, ensured to be Psoroptes-free, and released back into the SAM. Eleven radio-collared rams were translocated from the Red Rock Wildlife Area (RRWA) in New Mexico into the SAM range and monitored through 2002 to determine whether Psoroptes spp. mites were still in the environment. None of these sentinel rams acquired scabies during this period, and no additional native sheep were found to be present in the range. In 2002, 51 desert bighorn sheep were translocated into the SAM from the Kofa National Wildlife Refuge in Arizona (n = 20) and the RRWA in New Mexico (n = 31). Twenty-one bighorn sheep have died in the SAM since that time, but Psoroptes spp. mites have not been detected on any of these animals, nor have they been found on mule deer (Odocoileus hemionus) sampled since 2000. We conclude that psoroptic scabies is no longer present in the San Andres bighorn sheep population and that psoroptic scabies poses a minimal to nonexistent threat to the persistence of this population at this time.     

Optimized serodiagnosis of sheep fascioliasis by Fast-D protein liquid chromatography fractionation of Fasciola hepatica excretory-secretory antigens

Current methods for the serodiagnosis of sheep fascioliasis show suboptimal sensitivity, specificity, or both. With the aim of developing an improved method, we fractionated native Fasciola hepatica excretory-secretory antigens (ESAs) by size-exclusion FPLC (fast protein liquid chromatography) on a Superdex 75 HR 10/30 column and then tested the serodiagnostic value of the antigens contained in each one of the 4 peaks obtained (peaks I-IV). Serodiagnostic value was assessed using sera from sheep naturally infected with F. hepatica (group A); sera from the individuals of a fluke-free herd (most of which also had other intestinal nematodes, lung nematodes, Moniezia spp., and/or Cysticercus tenuicollis) sera from a fluke-free herd (group B); sera from lambs experimentally infected with 10-40 F. hepatica metacercariae (group C); and sera from uninfected control lambs (group D). Enzyme-linked immunosorbent assay (ELISA) with peak I or II as target antigens (and to a lesser extent with peak III as target) showed reactivity with negative sera, so that it was not possible to establish cutoff values discriminating infected and uninfected animals. In contrast, when peak IV was used as target, a low cutoff value of 0.235 optical density units (mean + 4 SD) discriminated infected and uninfected animals, with 100% sensitivity and 100% specificity. ELISA with peak IV as a target identified infected animals (even animals that had received only 10 metacercariae) within 3-5 wk of infection and subsequently throughout the rest of the 14-wk monitoring period. In Western blotting analysis, again only the antigens contained in peak IV (range 7-40 kDa, under reducing conditions) were specific for diagnosis of infected animals. These results indicate that molecular sieving of F. hepatica ESAs by this procedure is a fast, simple, reproducible way of obtaining antigens useful for serodiagnosis of sheep fascioliasis.     

Psoroptic scabies in Rocky Mountain bighorn sheep (Ovis canadensis canadensis) from Wyoming

Thirteen Rocky Mountain bighorn sheep (Ovis canadensis canadensis) with clinical signs of psoroptic scabies were captured in Wyoming. Signs included droopy ears, depilation on the head and neck, and exudate in the ears. Mites were identified as either Psoroptes cervinus or P. equi. Two ewes with scabies at the time of original capture had no clinical signs of mite infection 1 and 2 yr later.     

Spatial ecology of female bighorn sheep in a prairie landscape in Nebraska

Understanding the spatial ecology of small populations at the periphery of their range is important for identifying factors limiting population growth and developing sound management strategies. Bighorn sheep were reintroduced to Nebraska in 1981 and persist in a small population at the easternmost extent of the distribution of the species, where 1 of the 2 subpopulations is declining. Bighorn sheep space use and movement has been studied extensively in mountain and desert populations, but information is sparse from prairie populations in the Northern Great Plains. We investigated the spatial ecology of female bighorn sheep in Nebraska, USA, with global positioning system (GPS) telemetry. We tested the hypothesis that space use and movements would vary across seasons, years, and individuals but predicted that migration would involve relatively short distances in this translocated population (relative to native populations) occupying a fragmented landscape. Overall, females used smaller seasonal home ranges (3.3–7.8 km²) than most of those reported previously for the species and exhibited a high degree of variability in space use and movements across seasons, subpopulations, and individuals. Most females (92–100%) exhibited fidelity to their home ranges across seasons and years. Six females migrated between spatially distinct core lambing and winter ranges, although the distances (range = 7.9–8.7 km) and mean elevations (range = 31–41 m) between these core seasonal ranges were less than those reported for most native, migratory bighorn sheep populations. After accounting for variation in season, subpopulation, and years, home range size was positively associated with road density in both subpopulations (P < 0.001), suggesting that females incur greater energetic costs associated with greater space use in areas of higher fragmentation. Our results establish the basic spatial ecology of female bighorn sheep in Nebraska where their behavior appears to reflect the isolated nature of suitable habitat in this fragmented prairie landscape.     

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.     

Psoroptes sp. in two llamas (Lama glama) in Washington.

Psoroptes sp. mites were isolated from two llamas (Lama glama), a 4-mo-old male and its dam, in Washington. Mites were restricted to the ears and were responsible for head-shaking and incoordination. Infestations were treated successfully with a subcutaneous injection of ivermectin at 0.2 mg/kg of body weight and with 2 drops of ivermectin diluted in saline given topically in each ear. This is the first known report of Psoroptes sp. in llamas in the United States.     

Spatial and seasonal distribution of potential vectors of hemorrhagic disease viruses to peninsular bighorn sheep in the Santa Rosa Mountains of southern California.

Blood-feeding midges (Culicoides sp. and Leptoconops sp.) were sampled in the Santa Rosa Mountains, Riverside County, California (USA), to determine which species might be involved in the transmission of bluetongue and epizootic hemorrhagic disease viruses to peninsular bighorn sheep (Ovis canadensis cremnobates). Host-seeking midges were sampled with CO2-baited suction traps over a period of 30 mo. Nineteen species of Culicoides and seven of Leptoconops were collected. Five of the Culicoides sp. recovered are previously undescribed. The most abundant and widely distributed Culicoides sp. during spring (presumed virus transmission period to lambs) were C. (Selfia) brookmani, C. variipennis, C. (Avaritia) sp. (a new species near C. pusillus), and C. lahontan. Of these, C. brookmani (all elevations) and C. (Avaritia) sp. (elevations greater than 750 m) were common in the mountainous terrain inhabited by bighorn sheep. Culicoides variipennis, a vector of bluetongue virus in agricultural settings, and C. lahontan were numerous in sandy washes but were much less common in the mountains themselves. Leptoconops belkini and L. foulki were occasionally common in upper Deep Canyon in spring (April-June), while L. torrens was very abundant in the same area for 2 wk following heavy summer rains. Parity (an indicator of longevity and success in finding hosts and oviposition sites) in mountain areas was very low in C. variipennis (5%), low-moderate in C. (Avaritia) sp. (13%) and C. lahontan (21%), and relatively high in C. brookmani (40%). Vectorial capacity of Culicoides spp. for these hemorrhagic disease viruses is discussed, and it is suggested that species in addition to C. variipennis be considered as potential vectors of hemorrhagic disease viruses to desert bighorn sheep.     

Genetic characterization of Anaplasma ovis strains from bighorn sheep in Montana

Wildlife reservoir species and genetic diversity of Anaplasma ovis (Rickettsiales: Anaplasmataceae) have been poorly characterized. Bighorn sheep (Ovis canadensis), captured in Montana from December 2004 to January 2005, were tested for antibodies to Anaplasma spp.; the presence of A. ovis was determined by the characterization of major surface protein msp4 sequences. Anaplasma antibodies were detected in 25/180 (14%) sampled bighorn sheep and A. ovis msp4 sequences were amplified by polymerase chain reaction (PCR) and sequenced from 9/23 (39%) of seropositive animals. All animals were negative by PCR for the related pathogens, Anaplasma phagocytophilum and Anaplasma marginale. All msp4 sequences identified in the bighorn sheep were identical and corresponded to a single A. ovis genotype that was identical to a sheep isolate reported previously from Idaho. The finding of a single genotype of A. ovis in this wild herd of bighorn sheep was in contrast to the genetic diversity reported for A. marginale in cattle herds in the western United States and worldwide. These results demonstrated that bighorn sheep may be a wildlife reservoir of A. ovis in Montana.     

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.     

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.     

History, biology and control of sheep scab

Sheep scab is one of the oldest known diseases of sheep. It is caused by either of the mange mites Psoroptes ovis (Fig. 1 and cover) or Sarcoptes scabiei. P. ovis causes irritation so intense that sheep become preoccupied with scratching, cease to feed and rapidly become emaciated. It should be eradicated on welfare grounds alone (Fig. 2).     

Early immunodiagnosis of fasciolosis in ruminants using recombinant Fasciola hepatica cathepsin L-like protease

A diagnostic ELISA with recombinant Fasciola hepatica cathepsin L-like protease as antigen was developed to detect antibodies against F. hepatica in sheep and cattle. The recombinant cathepsin L-like protease was generated by functional expression of the cDNA from adult stage F. hepatica flukes in Saccharomyces cerevisiae. Specificity and sensitivity of the cathepsin L enzyme-linked immunosorbent assay (ELISA) was assessed using sera from sheep and calves experimentally or naturally mono-infected with F. hepatica and six-seven other parasites. The sensitivity of the cathepsin L ELISA for sheep and cattle sera was 99.1 and 100%, respectively. In the experimental setting with established mono-infections, the specificity of the cathepsin L ELISA was 98.5% for cattle sera and 96.5% for sheep sera. In experimentally infected cattle and sheep, the first detection of F. hepatica-specific antibodies appeared first between 5 and 7 weeks post-infection, but depended on the infectious dose of F. hepatica. In ELISA the detection preceded first detection of the infection based on egg counts and remained detectable till at least 23 weeks after a primary F. hepatica infection. Detection of Fasciola gigantica infections was similar to detection of F. hepatica. The first detection occurred at week 5 and signals persisted for at least 20 weeks. All sera from naturally F. hepatica infected sheep were seropositive in the cathepsin L-like ELISA. The relevance of this ELISA format was also evaluated using sera from naturally infected cattle in the Netherlands, Ecuador and Vietnam and compared with results from egg-counts. For the latter two endemic areas with mixed parasitic infections the 'apparent' sensitivity of the cathepsin L ELISA was calculated for all serum samples together to be 90.2%. The 'apparent' specificity under these conditions was calculated to be 75.3%. In cattle, the cathepsin L ELISA was superior to the concurrently evaluated peptide ELISA format using a single epitope as the antigen both in controlled natural infections as well as in infections in endemic areas. The present ELISA-format contributes a relatively sensitive and reliable tool for the early serodiagnosis of bovine and ovine fasciolosis.     

Evaluating apparent competition in limiting the recovery of an endangered ungulate

Predation can disproportionately affect endangered prey populations when generalist predators are numerically linked to more abundant primary prey. Apparent competition, the term for this phenomenon, has been increasingly implicated in the declines of endangered prey populations. We examined the potential for apparent competition to limit the recovery of Sierra Nevada bighorn sheep (Ovis canadensis sierrae), an endangered subspecies under the US Endangered Species Act. Using a combination of location, demographic, and habitat data, we assessed whether cougar (Puma concolor) predation on endangered bighorn sheep was a consequence of their winter range overlap with abundant mule deer (Odocoileus hemionus). Consistent with the apparent competition hypothesis, bighorn sheep populations with higher spatial overlap with deer exhibited higher rates of cougar predation which had additive effects on adult survival. Bighorn sheep killed by cougars were primarily located within deer winter ranges, even though those areas constituted only a portion of the bighorn sheep winter ranges. We suspect that variation in sympatry between bighorn sheep and deer populations was largely driven by differences in habitat selection among bighorn sheep herds. Indeed, bighorn sheep herds that experienced the highest rates of predation and the greatest spatial overlap with deer also exhibited the strongest selection for low elevation habitat. Although predator-mediated apparent competition may limit some populations of bighorn sheep, it is not the primary factor limiting all populations, suggesting that the dynamics of different herds are highly idiosyncratic. Management plans for endangered species should consider the spatial distributions of key competitors and predators to reduce the potential for apparent competition to hijack conservation success.     

Pathologic changes and microorganisms found in bighorn sheep during a stress-related die-off

An all-age die-off of Rocky Mountain bighorn sheep (Ovis c. canadensis Shaw) occurred from late October 1980 through March 1981 in Waterton Canyon, Colorado, with a loss of 75 to 85% of the sheep. The cause of death was a subacute to chronic bronchopneumonia and the primary etiologic agents isolated from the respiratory system were a Pasteurella sp., P. multocida, Corynebacterium pyogenes, and Protostrongylus stilesi Dikmans, 1931. The underlying predisposing factors that initiated this die-off were believed to be related to multiple chronic environmental stressors associated with the building of a dam which included human contact, vehicular traffic, atmospheric dust, noise and harassment. The die-off was succeeded by a 100% lamb mortality the following summer and a 67% lamb mortality the next two summers. The pneumonia found in these lambs was similar to that found in adult sheep during the previous die-off, except that mature lungworms were absent.     

Ecology of gastropod and bighorn sheep hosts of lungworm on isolated, semiarid mountain ranges in Utah, USA

Isolated, nonmigratory populations of bighorn sheep (Ovis canadensis) may experience high exposure to lungworms (Protostrongylus spp.) through a build-up of fecal material. However, semiarid climates may hinder lungworm transmission by limiting terrestrial gastropods, the intermediate hosts. We assessed potential for lungworm transmission, documented occurrence of transmission, and identified habitat types where transmission was likely to occur on ranges of two recently introduced populations of bighorn sheep in northern Utah. Gastropods were collected weekly on Antelope Island and the Newfoundland Mountains, May-August 2001-02, from each of the four major habitat types (riparian, rock, desert shrub, and grass). Distribution of 113 bighorn sheep groups was observed, and 421 fecal pellet groups were collected to estimate lungworm levels. A total of 1,595 gastropods representing five genera were collected from both ranges. Vallonia made up 85% of all gastropods collected. Of 980 gastropods collected on Antelope Island in 2002, only Vallonia were found infected with protostrongylid-type larvae (10 of 980=1%). Lungworm prevalence in bighorn fecal samples was 97% on Antelope Island and 90% on the Newfoundland Mountains. Lungworm prevalence in lambs indicated lungworm transmission was occurring on Antelope Island. Lungworm transmission was likely occurring in riparian habitat due to abundant gastropods, presence of infected gastropods, and reliance by bighorn sheep on few water sources. Differences in spatial distribution between ram and nursery groups may partly explain higher fecal larvae counts in nursery than in ram groups. We suggest lungworm levels in bighorn sheep on semiarid ranges may increase in dry years as bighorn sheep concentrate use on fewer perennial water sources.