Comparison of three fecal steroid metabolites for pregnancy detection used with single sampling in bighorn sheep (Ovis canadensis)

We conpared three fecal steroid metabolite assays for their usefulness in detecting pregnalcy among free-ranging Rocky Mountain bighorn sheep (Ovis canadensis canadensis) from Bighorn Canyon National Recreation Area, Wyoming and Montana (USA) and captive bighorn ewes at ZooMontana in Billings, Montana. Fecal samples were collected from 11 free-ranging, radio-collared bighorn ewes in late January-May 2001 and from 20 free-ranging, radio-collared ewes in late March to mid-May 2002. Free-ranging ewes were monitored the following spring to determine whether or not they lambed. In addition, two captive ewes were studied at ZooMontana. With three exceptions, free-ranging bighorn ewes that produced lambs had nonspecific progesterone metabolite (iPdG) levels of >1800 ng/g feces and iPdG levels >7000 ng/gm feces when samples were collected between early March and mid-May. Samples collected earlier in the year were inconclusive. One false negative was suspected to be the result of sample collection error. Of the captive ewes, nonspecific pregnanediol-3alpha-glucuronide (PdG) and iPdG followed a predictable curve over the course of the 180-day pregnancies. We conclude that estrone conjugates are not useful in diagnosing pregnancy; however, fecal steroid analysis of PdG and iPdG can be used to accurately determine pregnancy and reproductive function in bighorn sheep. This holds great potential as a noninvasive technique for understanding the role of reproductive disease in wild bighom 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.     

Fecal counts of lungworm larvae and reproductive effort in bighorn sheep, Ovis canadensis

Because parasite resistance and reproduction require metabolic resources, life-history models predict a tradeoff between current reproduction and parasite load. These tradeoffs have been widely studied in birds, but few studies have been conducted on mammals. We monitored lungworm ( Protostrongylus spp.) larvae counts in bighorn sheep ( Ovis canadensis ) over four years to examine how individual differences in fecal output of lungworm larvae (LPG) by yearlings and adults were affected by season, sex, body mass, age and reproductive effort. We also compared lamb mass at six months and LPG. Overall, we found that LPG varies seasonally, peaking in females prior to lambing and in males during the rut. Age had no effect on LPG for either sex. During autumn, we found no effect of age or mass on LPG for sheep one year and older. Lamb body size or sex did not affect LPG. Females that weaned a lamb had higher counts than females that did not produce a lamb or females whose lamb died during summer. For rams, social rank and testosterone levels were not related to LPG but LPG increased with time spent searching for estrous ewes during the rut. Our results suggest a tradeoff between parasite resistance and reproductive effort in bighorn sheep of both sexes.     

Influence of rainfall on lungworm infections in bighorn sheep.

From 1959 through 1968, lungs from 124 Rocky Mountain bighorn sheep (Ovis c. canadensis) from the Sun River herd in western Montana were examined for lungworm infections. All lungs were infected with Protostrongylus stilesi and 104 (84%) contained concurrent infections of P. stilesi and P. rushi. Significant correlations were observed between levels of lungworm infection and total rainfall during April, May, and June of each year. An explanation of this in terms of terrestrial snail (intermediate host) populations and a suggestion for the possible use of these data in developing a predictive model for forecasting lungworm levels for use in in bighorn sheep management are given.     

Evaluation of management treatments intended to increase lamb recruitment in a bighorn sheep herd

We administered a suite of treatments to a herd of Rocky Mountain bighorn sheep (Ovis canadensis canadensis) that was experiencing poor lamb recruitment and showing signs of respiratory disease. Despite 3 yr of treatment with various combinations of anthelmentics, antibiotics, vaccines, and hyperimmune serum products, recruitment was not improved.     

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.     

Comparison of Endocrine Response to Stress Between Captive-Raised and Wild-Caught Bighorn Sheep

ABSTRACT Stress hormones in Rocky Mountain bighorn sheep (Ovis canadensis canadensis), produced in response to environmental changes, road development, or high population density, may impact their immune systems to a threshold level that predisposes them to periodic, large-scale mortality. We compared the stress response to a novel environmental situation and repeated handling between bighorn sheep born and raised in captivity (CR) and bighorn sheep born in the wild (WC) and brought into captivity. We measured plasma epinephrine, norepinephrine, cortisol, and fecal glucocorticoid metabolites (FGM). Three weeks after each group's arrival we used a one-time drop-net event to elicit an acute stress response, and we collected blood samples from each sheep over 35 minutes, as well as one fecal sample. We collected blood and fecal samples from both groups on 7 other occasions over the subsequent 6 months. We also collected fecal samples from the pen at approximately 24-hour intervals for 3 days following every handling event to monitor the stress response to handling. We found that CR sheep had a stronger autonomic nervous system response than WC sheep, as measured by epinephrine and norepinephrine levels, but we found a very similar hypothalamic-pituitary–adrenal axis (HPA) response, measured by cortisol levels, to the acute stress event of a drop-net restraint. We also found that once the WC sheep had acclimated, as indicated by the return to the initial baseline FGM levels within 12 weeks, the CR and WC groups' HPA responses to sampling events were not significantly different from one another. Fecal samples can provide a noninvasive mechanism for managers to monitor baseline FGM for a given herd. Using long-term monitoring of FGM rather than values from a single point in time may allow managers to correlate these levels to outside influences on the herd and better understand the impacts of management changes, population density, or increased human developments on the health of the sheep population.     

Freemartinism in a captive herd of Rocky Mountain bighorn sheep (Ovis canadensis).

Freemartinism in two animals from a captive herd of Rocky Mountain bighorn sheep (Ovis canadensis) at the Denver Zoological Gardens (Denver, Colorado, USA) is described. A young ewe had female external genitalia, a masculine appearance, and demonstrated male behavior as she matured. Another ewe with female external genitalia died as a yearling. Necropsy revealed a non-patent vagina and internal male genitalia. Both females were chimeric with karyotypes containing XX and XY sex chromosomes.     

Experimental infections of free-ranging Rocky Mountain bighorn sheep with lungworms (Protostrongylus spp.; Nematoda: Protostrongylidae)

Twelve free-ranging Rocky Mountain bighorn lambs (Ovis canadensis canadensis), each exposed experimentally to 125-1,000 infective third-stage larvae of Protostrongylus stilesi and P. rushi, shed significantly more first-stage larvae in their feces than did control lambs, but showed no clinical signs of illness and had equivalent summer and overwinter survival as control lambs. Two adult ewes, each exposed to 925 infective larvae, showed no increase in numbers of first-stage larvae in their feces; both survived at least 14 mo postexposure. Experimentally exposed lambs did not differ from control lambs in numbers of larvae in their feces in the following summer. Three experimental lambs had 313-402 adult P. stilesi and 0-97 adult P. rushi on necropsy; two control lambs had 255 and 270 P. stilesi and no P. rushi. The presence of these numbers of lungworms did not appear to be sufficient to precipitate lungworm pneumonia in bighorn lambs under the conditions of this study.     

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.     

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.     

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.     

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.     

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.     

Isolation of a chlamydial agent from Rocky Mountain bighorn sheep

A total of 53 clinical specimens from both healthy and diseased Rocky Mountain bighorn sheep (Ovis canadensis) were examined for Chlamydia. An agent consistently lethal for chicken embryos was recovered from a nasal swab taken from a normal ewe. This agent, designated BHS-15, possesses antigens which fix complement in the presence of anti-chlamydial serum, is susceptible to chlortetracycline, and is resistant to sodium sulfadiazine and streptomycin. Attempts to culture the isolate in quality control media, including blood agar, thioglycolate broth, and PPLO broth and agar were unsuccessful. A recommendation is made for classification of agent BHS-15 as a member of the species Chlamydia psittaci. The possible relationship of the isolate to the pneumonia complex in bighorn sheep is discussed.     

Intrasexual Competition and Mate Choice in Rocky Mountain Bighorn Sheep

Rocky Mountain bighorn (Ovis canadensis canadensis) rams employed three distinct mating tactics. When tending, rams defended single estrous ewes. In coursing, rams forced copulations with defended ewes, and, in blocking, rams sequestered ewes from more dominant rams. Ewes utilized a traditional area when tended, attempted to escape to this area when blocked, and resisted coursing ram attempts to force copulations. Between-year variation in the dispersion of estrous ewes about the tending area strongly influenced the consort and probably mating success of dominant rams. Thus, ewe spatial predictability during estrus — achieved by clustering in tended estrus and resisting blocking rams — appears to be an important mechanism of mate choice in this species. Ewes apparently did not gain material or risk-related benefits by mating dominant rams. That such males provide ewes with “good genes” is an attractive remaining possibility.     

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.     

Prevalence of Dictyocaulus viviparus infection in Rocky Mountain elk in Teton County, Wyoming.

Dictyocaulus viviparus infections in Rocky Mountain elk (Cervus canadensis of Teton Countywere surveyed by fecal analyses during spring, summer and winter and by fecal analyses and necropsies during fall hunting seasons, 1968-1973. Prevalance of the lungworms was relatively high: 32-70% during the spring; slightly lower, 30-47%, during the summer; 21-39% in the fall; and declined to the annual low of 8-19% during the winter. Conversely, elk summering on Big Game Ridge showed an increase in prevalance of D. viviparus from 1969 to 1973. Decreases in prevalance of lungworms were noted on the National Elk Refuge at Jackson after management changes were effected in 1971.     

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.     

Occurrence, diagnosis, and strain typing of Mycobacterium avium subspecies paratuberculosis infection in Rocky Mountain bighorn sheep (Ovis canadensis canadensis) in southwestern Alberta

The role that wildlife may play in the transmission of Mycobacterium avium subspecies paratuberculosis (Map), the causative agent of Johne's disease (JD), and the potential consequences of infection in these populations are being given increasing consideration. A yearling male Rocky Mountain bighorn sheep (Ovis canadensis canadensis) from southwestern Alberta, Canada, was found infected with Map in August 2009. Clinical signs of emaciation and diarrhea and histologic findings of diffuse granulomatous enteritis of the distal ileum, lymphadenitis of the mesenteric lymph nodes, and lymphangitis of the ileum were similar to previously described cases of JD in bighorn sheep. Infection with Map was confirmed by bacterial isolation through fecal culture, acid-fast staining, and polymerase chain reaction (PCR) of IS900. The Map1506 gene was sequenced, and the isolate was identified as a Cattle (Type II) strain. In a follow-up herd-level survey, three of 44 fecal samples (7%) from individual bighorn sheep from the same herd as the index case were PCR-positive and identified as Type II Map strains. Twenty-five samples from a distant bighorn population were negative. Additional strain typing of the isolates from the index case and the positive fecal samples was done by sequencing three discriminatory short sequence repeat (SSR) regions. All four SSR profiles differed from one another, suggesting multiple introductions or a long-existing circulation of Map within this bighorn population. Detailed molecular analyses are essential for understanding and managing diseases at the wildlife-livestock interface.