Protostrongylid parasites and pneumonia in captive and wild thinhorn sheep (Ovis dalli)

We describe health significance of protostrongylid parasites (Parelaphostrongylus odocoilei and Protostrongylus stilesi) and other respiratory pathogens in more than 50 naturally infected Dall's sheep (Ovis dalli dalli) from the Mackenzie Mountains, Northwest Territories (1998-2002) as well as in three Stone's sheep (O. d. stonei) experimentally infected with P. odocoilei (2000-2002). Histological lesions in the brain and distribution of P. odocoilei in the muscles of experimentally and naturally infected sheep were consistent with a previously hypothesized "central nervous system to muscle" pattern of migration for P. odocoilei. Dimensions of granulomas associated with eggs of P. odocoilei and density of protostrongylid eggs and larvae in the cranial lung correlated with intensity of larvae in feces, and all varied with season of collection. Prevalence of P. stilesi based on the presence of larvae in feces underestimated true prevalence (based on examination of lungs) in wild Dall's sheep collected in summer and fall. Similarly, counts of both types of protostrongylid larvae in feces were unreliable indicators of parasitic infection in wild Dall's sheep with concomitant bacterial pneumonia associated with Arcanobacterium pyogenes, Pasteurella sp., and Mannheimia sp. Diffuse, interstitial pneumonia due to P. odocoilei led to fatal pulmonary hemorrhage and edema after exertion in one experimentally infected Stone's sheep and one naturally infected Dall's sheep. Bacterial and verminous pneumonia associated with pathogens endemic in wild Dall's sheep in the Mackenzie Mountains caused sporadic mortalities. There was no evidence of respiratory viruses or bacterial strains associated with domestic ruminants, from which this population of wild sheep has been historically isolated.     

Development and pathogenesis of Parelaphostrongylus odocoilei (nematoda: protostrongylidae) in experimentally infected thinhorn sheep (Ovis dalli )

Recently, the protostrongylid nematode Parelaphostrongylus odocoilei has been reported in a new host species, thinhorn sheep (Ovis dalli). For the first time, we completed the life cycle of P. odocoilei in three Stone's sheep (O. dalli stonei) and two thinhorn hybrids (O. dalli stonei x O. dalli dalli), each infected with 200 third-stage larvae from slugs (Deroceras laeve). The prepatent period ranged from 68 days to 74 days, and shedding of first-stage larvae (L1) peaked at >10,000 L1 per gram of feces between 90 and 110 days postinfection. A total of 75, 27, and 14 adult P. odocoilei were recovered from skeletal muscles of three Stone's sheep. Starting in the prepatent period, all infected sheep lost weight and developed peripheral eosinophilia. At 2 wk before patency, two thinhorn hybrids developed neurologic signs (hind end ataxia, loss of conscious proprioception, and hyperesthesia) that resolved at patency. Eosinophilic pleocytosis and antibody to Parelaphostrongylus spp. were detected in the cerebrospinal fluid of the affected sheep, suggesting that the migration route of the "muscleworm" P. odocoilei may involve the central nervous system. Twenty days after treatment with ivermectin, neurologic signs recurred and larval shedding ceased in one infected thinhorn hybrid, whereas multiple treatments transiently suppressed but did not eliminate larval shedding in the other. During patency, two Stone's sheep with numerous eggs and larvae of P. odocoilei in the lungs died of respiratory failure following anesthesia or exertion. Parelaphostrongylus odocoilei has widespread geographic distribution, high prevalence, the possibility of causing neurologic and respiratory disease, resistance to treatment, and may constitute a significant emerging disease risk for thinhorn sheep.     

Muskox lungworm (Umingmakstrongylus pallikuukensis) does not establish in experimentally exposed thinhorn sheep (Ovis dalli)

Muskoxen (Ovibos moschlatus moschatus) on the northwestern mainland of Nunavut and Northwest Territories, Canada, are infected with the protostrongylid lungworm, Umingmaksrongylus pallikuuhkensis. The geographic range of this muskox population is expanding to the south and west, and it is anticipated that these animals will eventually become sympatric with Dall's sheep (Ovis dalli dalli) in the Mackenzie and Richardson Mountains. To address the concern of wildlife managers that U. pallikulkensis may infect and adversely affect Dalls sheep, four Dalls/Stone's (Ovis dalli stonei) hybrid lambs and one adult muskox (Ovibos moschatus wardi) were each given 100 third-stage larvae of U. pallikuukensis. All animals were intensively monitored for 9 mo postinfection (PI) using clinical examinations, fecal analyses, hematology, blood chiemistry and medical imaging. No first-stage larvae of U. pallikuinkensis were recovered from the lambs and monitoring revealed nio evidence that the parasite had established in any of these animals. First-stage larvae were found in the feces of the muskox beginning at 94 days PI, and typical parasite cysts were visible in lung radiographs at 188 days PI. This study addresses an important management and wildlife health issue associated with the potential for host-switching of pathogens and indicates that it is improbable that thinhorn sheep are suitable hosts for U. pallikuukensis.     

Geographic distribution of the muscle-dwelling nematode Parelaphostrongylus odocoilei in North America, using molecular identification of first-stage larvae

Molecular identification of dorsal-spined larvae (DSL) from fecal samples indicates that the protostrongylid parasite Parelaphostrongylus odocoilei occupies a broader geographic range in western North America than has been previously reported. We analyzed 2,124 fecal samples at 29 locations from thinhorn sheep (Ovis dalli dalli and O. d. stonei), bighorn sheep (Ovis canadensis canadensis and O. c. californiana), mountain goats (Oreamnos americanus), woodland caribou (Rangifer tarandus caribou), mule deer (Odocoileus hemionus hemionus), and black-tailed deer (O. h. columbianus). The DSL were recovered from populations of thinhorn sheep south, but not north, of the Arctic Circle, and they were not recovered from any of the bighorn sheep populations that we examined. In total, DSL were recovered from 20 locations in the United States and Canada (Alaska, Yukon Territory, Northwest Territories, British Columbia, Alberta, and California). The DSL were identified as P. odocoilei by comparing sequences of the second internal transcribed spacer (ITS2) region of ribosomal RNA among 9 protostrongylid species validated by adult comparative morphology. The ITS2 sequences were markedly different between Parelaphostrongylus and other protostrongylid genera. Smaller fixed differences served as diagnostic markers for the 3 species of Parelaphostrongylus. The ITS2 sequences (n = 60) of P. odocoilei were strongly conserved across its broad geographic range from California to Alaska. Polymorphism at 5 nucleotide positions was consistent with multiple copies of the ITS2 within individual specimens of P. odocoilei. This work combines extensive fecal surveys, comparative morphology, and molecular diagnostic techniques to describe comprehensively the host associations and geographic distribution of a parasitic helminth.     

Bionomics of larvae of Parelaphostrongylus odocoilei (Nematoda: Protostrongylidae) in experimentally infected gastropod intermediate hosts

Parelaphostrongylus odocoilei is a protostrongylid parasite that has recently been recognized at several locations in sub-Arctic, but not Arctic, North America. We investigated factors that may determine the distribution of P. odocoilei, including suitable gastropod intermediate hosts, temperature requirements for larval development in gastropods, and larval emergence facilitating overwinter transmission. We collected and experimentally infected gastropods from a site in the sub-Arctic where P. odocoilei is at the northern limit of its distribution. Deroceras laeve, Catinella sp., and Euconulus cf fulvus, but not members of the Pupillidae, were suitable intermediate hosts. We describe bionomics of larvae of P. odocoilei in D. laeve and Catinella sp. Infective larvae emerged from all slugs (D. laeve) and 60% of Catinella sp. snails, and emergence from D. laeve was intensity dependent. Emerged infective larvae survived up to 6 mo under conditions approximating that of the subnivean environment. In D. laeve, there was a direct relationship between temperature and development rate of larvae of P. odocoilei. Larvae of P. odocoilei did not develop to infective stage below the theoretical threshold (8.5 C), and required a minimum of 163 degree days to complete development. These developmental parameters can be incorporated into a model to predict larval development in the field. Knowledge of the factors influencing larval bionomics provides the foundation for predicting temporal and spatial patterns of parasite distribution, abundance, and transmission.     

Emergence of third-stage larvae of Umingmakstrongylus pallikuukensis from three gastropod intermediate host species

We investigated the emergence of third-stage larvae (L3) of Umingmakstrongylus pallikuukensis from the slugs Deroceras laeve, Deroceras reticulatum, and the snail Catinella sp. in the laboratory and from D. laeve on the tundra. Third-stage larvae emerged from 8 of 8 D. laeve and 8 of 8 D. reticulatum housed at 20 C in darkness and from 9 of 10 D. laeve and 5 of 5 Catinella sp. housed at 21 C with 10-12 hr of light/day. Larvae emerged from D. laeve and D. reticulatum over a wide range of infection intensities (2-179 and 20-65, respectively), and the patterns of emergence were independent of intensity. The majority of the L3 emerged from most of the Deroceras spp. by 58 or 60 days postinfection (PI). Lower rates of emergence were observed from Catinella sp. Larvae emerged from D. laeve on the tundra by 10 wk PI and were recovered from the vegetation in some experimental enclosures the following year. Third-stage larvae survived in tap and distilled water at 0-4 C for 13 mo. Emergence of L3 of U. pallikuukensis from the intermediate host may increase the temporal and spatial availability of L3 and enhance their survival and transmission.     

Population genetic structure of North American thinhorn sheep (Ovis dalli)

The thinhorn sheep (Ovis dalli ssp.) provides a rare example of a North American large mammal that occupies most of its native range and maintains close to ancestral population size. There are currently two recognized subspecies, Dall's sheep (O. d. dalli) and Stone's sheep (O. d. stonei), the validity of which remains uncertain. We investigated the spatial genetic structure of thinhorn sheep populations representing both subspecies by genotyping individuals (n = 919) from across the species range at 12 variable microsatellite loci. We found high levels of genetic diversity within (HE = 0.722) and significant genetic structure among the 24 sampled areas (FST = 0.160). Genetic distance measures and Bayesian clustering analyses revealed the presence of at least eight subpopulations that are delineated by mountain range topology. A strong overall pattern of isolation-by-distance is evident across the sampling range (r = 0.75, P < 0.001) suggesting limited dispersal and extensive philopatry. Partial Mantel tests of this relationship showed mountain range distinctions represent significant barriers to gene flow (P = 0.0001), supporting the Bayesian analyses. Genetic structure was more strongly pronounced in southern Yukon and Alaska than elsewhere. We also show evidence for genetic differences between the two currently recognized thinhorn subspecies.     

Failure to maintain interspecific pregnancy after transfer of Dall's sheep embryos to domestic ewes

Over a 3-year period, 32 Dall's sheep (Ovis dalli dalli) embryos were transferred into 24 domestic sheep (O. aries) recipients and 4 were transferred into 2 Dall's sheep recipients. In the first year, none of the 10 O. aries recipients was diagnosed pregnant. In the following 2 years, 9 (37%) of the domestic sheep recipients were pregnant on Day 18, 8 (33%) on Day 40, 6 (25%) on Day 90 and 4 (16%) on Day 120; 1 aborted at Day 125 and another at Day 145. Pregnancies were established only in ewes that had previously been recipients of Dall's sheep embryos. The 2 remaining pregnant sheep were treated with progesterone from Day 125 until the fetuses were determined to be dead at Day 145. Both of the Dall's sheep recipients (Year 2) established pregnancies; 1 live Dall's sheep lamb was born 174 days after mating. No differences in serum progesterone, oestrone, prostaglandin F-2 alpha metabolites or cortisol concentrations could be detected during pregnancy between recipients carrying Dall's sheep embryos, recipients receiving progesterone treatment or domestic ewes carrying domestic sheep pregnancies. Six fetuses were necropsied (1 at Day 125 and 5 at Day 145-146): all fetuses were premature and had various degrees of hydranencephaly. No significant differences were found when cotyledon numbers were compared among domestic ewes carrying Dall's sheep lambs. Dall's sheep ewes lambing naturally and domestic ewes lambing naturally. These results demonstrate that the transfer of Dall's sheep embryos to domestic ewes results in the establishment but subsequent loss of pregnancy and that these losses occur throughout gestation.     

Bighorn sheep, a new host record for Parelaphostrongylus odocoilei (Nematoda: Protostrongylidae)

Larval nematodes with a dorsal spine on the tail were recovered from fecal samples of California bighorn sheep (Ovis canadensis californiana) in northeastern Washington State, USA. The identity of these dorsal-spined larvae (DSL) was established by single-strand conformation polymorphism (SSCP) analyses of a partial fragment of the first internal transcribed spacer of the ribosomal DNA. The SSCP profiles of individual DSL from bighorn sheep were compared with those of DSL of five protostrongylid species (Parelaphostrongylus andersoni, P odocoilei, P. tenuis, Elaphostrongylus rangiferi, and Muellerius capillaris) but were identical to only those of P. odocoilei. This study represents the first confirmed identification of P. odocoilei in bighorn sheep.     

Lumpy jaw in wild sheep and its evolutionary implications

The distribution and prevalence of mandibular osteomyelitis, lumpy jaw, and other dental anomalies in wild sheep were investigated and their biological and evolutionary implications were assessed. Our survey was based on 3,363 mandibles of wild sheep and 1,028 from domesticated varieties. Lumpy jaw is widespread in wild sheep of North America, but it is rare or absent in wild sheep from Eurasia. Among the subspecies of Ovis spp. in North American, the thinhorn sheep (Ovis dalli) were the most seriously impacted, with a prevalence in Dall's sheep (O. dalli dalli) of 23.3% and 29.3% in Stone's sheep (O. dalli stonei). Among the bighorns (O. canadensis), the Rocky Mountain subspecies (O. canadensis canadensis) had a higher rate (12.1%) than other subspecies. Lumpy jaw was not documented in the desert sheep of Baja California (O. canadensis cremnobates, O. canadensis weemsii). Based on data from affected thinhorn sheep, it appears there is an inverse relationship between age of a subspecies in a long term evolutionary context and susceptibility to lumpy jaw. In Eurasian wild sheep lumpy jaw is rare or absent with prevalences ranging from 0 to 7.1% among suspecies, and in domesticated breeds the prevalence averaged 5.0%. The impact of lumpy jaw on different age classes or longevity is equivocal, although females are more susceptible than males. Lumpy jaw appears to effect horn development in males.     

Serologic and experimental investigations of contagious ecthyma in Alaska

Serologic evidence of contagious ecthyma (CE) was found in domestic sheep (Ovis aries), domestic goats (Capra hircus), Dall sheep (Ovis dalli), and musk-ox (Ovibos moschatus) in Alaska. A moose (Alces alces) calf and a caribou (Rangifer tarandus) fawn were susceptible to experimental infection and both developed antibody titers as a result. CE virus was isolated from lesions of Dall sheep which were involved in a natural outbreak of the disease.     

An epizootic of Mycoplasma ovipneumoniae infection in captive Dall's sheep (Ovis dalli dalli)

In late spring of 1986, 10 of 23 Dall's sheep (Ovis dalli dalli) at the Metropolitan Toronto Zoo were moved to a new exhibit, where all developed severe respiratory signs refractory to anthelmintic and antibiotic therapy. In July, two animals died with chronic active bronch-pneumonia, and a third was euthanized because of pneumonia several months later. Bacteria were not isolated from the lungs of the first, steptococci and Pasteurella hemolytica were isolated from the other two, respectively; Mycoplasma ovipneumoniae was isolated from both. Pulmonary lesions in all three sheep were consistent with Mycoplasma sp. infection. Nasal swabs of the remaining animals yielded no consistent bacterial isolates; however, four of eight sheep were positive for M. ovipneumoniae. Viral cultures yielded an as yet unidentified herpesvirus. Sheep in the original and new herds had no serologic titers to parainfluenza-3, equine viral rhinopneumonitis, or infectious bovine rhinotracheitis, and had variable titers against bovine respiratory syncytial virus. No titers against M. ovipneumoniae were present in 13 sheep still in the original exhibit, but titers varied from 1:32 to 1:256 in eight pneumonic sheep. Sera taken from three sheep before or early in the outbreak were all negative for antibody to M. ovipneumoniae. Two of the affected Dall's sheep had been in contact with domestic sheep in the winter of 1985-1986, and M. ovipneumoniae was subsequently cultured from the domestic flock. Exposure to a new pathogen, and environmental and social stress in a new exhibit may have resulted in this severe disease in Dall's sheep.     

Dall's sheep horn growth and harvest management in the Mackenzie Mountains,Northwest Territories,Canada

Across most of their native North American range, the horns of mountain sheep (Ovis spp.) males are getting smaller, a pattern attributed to selective hunting pressure. We measured the horns of 755 Dall's sheep males (Ovis dalli dalli) in the southern Mackenzie Mountains, Northwest Territories, between 2002 and 2017. For each male, we measured the circumference and length of each annulus for the right horn and calculated horn volume for each year. We examined changes in horn size in 4 different outfitter areas, using age at harvest as a covariate. Hunting pressure across years in the study area was consistently low, and this population did not experience the decline in horn size observed in several other mountain sheep populations in Canada. Over the 16-year period, the average horn volume of harvested males was stable and even increased in 1 outfitter area. Local management of Dall's sheep delivered independently by the guide outfitters in the Mackenzie Mountains appears to contribute to maintaining a population of males that has not been adversely affected by strong selective hunting pressure. The resilience of this management strategy may be challenged by environmental changes associated with rapid warming in northern mountain environments.     

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.     

Phylogenetic analysis of snow sheep (Ovis nivicola) and closely related taxa

Based on mitochondrial cytochrome b gene sequence analysis, the history of true sheep (Ovis) began approximately 3.12 million years ago (MYA). The evolution of Ovis resulted in three generally accepted genetic groups: Argaliforms, Moufloniforms, and Pachyceriforms. The Pachyceriforms of the subgenus Pachyceros comprise the thin-horn sheep Ovis nivicola (snow sheep), Ovis dalli (Dall and Stone sheep), and Ovis canadensis (Rocky Mountain and desert bighorn). North America wild sheep (O. canadensis and O. dalli) evolved separately from Eurasian wild sheep and diverged from each other about 1.41 MYA. Ancestral stock that gave rise to snow sheep, Moufloniforms, and Argaliforms occurred 2.3 MYA, which then gave rise to two different extant lines of snow sheep that diverged from each other about 1.96 MYA. The more recent nivicola line is genetically closer to the North American wild sheep and may represent a close association during the refugium when Alaska and Siberia were connected by the Bering land bridge. The earlier period of evolution of the Pachyceriforms suggests they may have first evolved in Eurasia, the oldest ancestor then giving rise to North American wild sheep, and that a canadensis-like ancestor most likely gave rise to nivicola. Cytogenetic analysis further validates that the standard diploid number for modern nivicola is 52.     

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.     

Eimeria dalli sp. n. (Protozoa: Eimeriidae) from Dall Sheep Ovis dalli

Eimeria dalli sp. n. is described from fecal samples collected from Dall sheep, Ovis dalli, from the Kenai Peninsula, Alaska. The oocysts are spherical or subspherical with mean dimensions of 43.7 × 37.4 μm. The outer oocyst wall is rough and irregular. No micropyle, micropylar cap or residuum was observed. Sporocysts were elongate ovoid with mean dimensions of 19.0 × 10.7 μm. Stieda bodies were not discernible.     

Serum antibody prevalence of malignant catarrhal fever viruses in seven wildlife species from Alaska

Blood samples were collected from seven species of free-ranging ungulates in Alaska. Sera were tested for evidence of exposure to malignant catarrhal fever viruses (MCFV) by means of a competitive enzyme-linked immunosorbent assay. Antibody prevalences were as follows: muskox (Ovibos moschatus) 100 positive samples of 104 tested (96%); Dall sheep (Ovis dalli) 212 of 222 (95%); elk (Cervus elaphus) 14 of 51 (27%); bison (Bison bison) 34 of 197 (17%); caribou (Rangifer tarandus) nine of 232 (4%); Sitka black-tailed deer (Odocoileus hemionus sitkensis) one of 49 (2%); and moose (Alces alces) three of 219 (1%). Antibody prevalence in a bison population from the Interior was stable over a 5 yr period. These results indicate that at least one virus in the MCF group is enzootic in Dall sheep and muskox in Alaska. Lower antibody prevalences in the other species in this survey suggest that MCFV are latent or subclinical in these free-ranging ruminants. Whole blood samples were collected from 14 Dall sheep and subjected to a polymerase chain reaction assay. Fragments of ovine herpesvirus-2 DNA were detected in six of the samples. The significance of these findings for the health of free-ranging ungulates in Alaska is unknown.     

Low Temperatures Suppress Growth of the Ciliate Parasite, Tetrahymena rostrata , and Pathogenicity to Field Slugs, Deroceras reticulatum

The slug parasitic ciliate protozoan, Tetrahymena rostrata , was isolated from the slug Deroceras reticulatum and used in bioassays against D. reticulatum at 10 and 17oC and in in vitro growth experiments at a range of temperatures. The parasite reproduced, and significantly reduced the life span of juvenile D. reticulatum at 17oC, but not at 10oC.