Phenotypic plasticity and local adaptation in freeze tolerance: Implications for parasite dynamics in a changing world.

Marshallagia marshalli is a multi-host gastrointestinal nematode that infects a variety of artiodactyl species from temperate to Arctic latitudes. Eggs of Marshallagia are passed in host faeces and develop through three larval stages (L1, L2, and L3) in the environment. Although eggs normally hatch as L1s, they can also hatch as L3s. We hypothesised that this phenotypic plasticity in hatching behaviour may improve fitness in subzero and highly variable environments, and this may constitute an evolutionary advantage under current climate change scenarios. To test this, we first determined if the freeze tolerance of different free-living stages varied at different temperatures (-9 °C, -20 °C and -35 °C). We then investigated if there were differences in freeze tolerance of M. marshalli eggs sourced from three discrete, semi-isolated, populations of wild bighorn and thinhorn sheep living in western North America (latitudes: 40°N, 50°N, 64°N). The survival rates of eggs and L3s were significantly higher than L1s at -9 °C and -20 °C, and survival of all three stages decreased significantly with increasing freeze duration and decreasing temperature. The survival of unhatched L1s was significantly higher than the survival of hatched L1s. There was no evidence of local thermal adaptation in freeze tolerance among eggs from different locations. We conclude that developing to the L3 in the egg may result in a fitness advantage for M. marshalli, with the egg protecting the more vulnerable L1 under freezing conditions. This phenotypic plasticity in life-history traits of M. marshalli might be an important capacity, a potential exaptation capable of enhancing parasite fitness under temperature extremes.

Adaptations and phenotypic plasticity in developmental traits of Marshallagia marshalli.

Despite the economic, social and ecological importance of the ostertagiine abomasal nematode Marshallagia marshalli, little is known about its life history traits and its adaptations to cope with environmental extremes. Conserved species-specific traits can act as exaptations that may enhance parasite fitness in changing environments. Using a series of experiments, we revealed several unique adaptations of the free-living stages of M. marshalli that differ from other ostertagiines. Eggs were isolated from the feces of bighorn sheep (Ovis canadensis) from the Canadian Rocky Mountains and were cultured at different temperatures and with different media. Hatching occurred primarily as L1s in an advanced stage of development, morphologically very similar to a L2. When cultured at 20 °C, however, 2.86% of eggs hatched as L3, with this phenomenon being significantly more common at higher temperatures, peaking at 30 °C with 28.95% of eggs hatching as L3s. After hatching, free-living larvae of M. marshalli did not feed nor grow as they matured from L1 to infective L3. These life history traits seem to be adaptations to cope with the extreme environmental conditions that Marshallagia faces across its extensive latitudinal distribution in North America and Eurasia. In order to refine the predictions of parasite dynamics under scenarios of a changing climate, basic life history traits and temperature-dependent phenotypic behaviour should be incorporated into models for parasite biology.

MOLECULAR SURVEILLANCE FOR BARTONELLA, BORRELIA, AND RICKETTSIA SPECIES IN TICKS FROM DESERT BIGHORN SHEEP ( OVIS CANADENSIS) AND MULE DEER ( ODOCOILEUS HEMIONUS) IN SOUTHERN CALIFORNIA, USA.

: Ticks (Acari: Ixodidae) were collected from 44 desert bighorn sheep ( Ovis canadensis) and 10 mule deer ( Odocoileus hemionus) in southern California, US during health inspections in 2015-16. Specimens were identified and screened by PCR analysis to determine the presence and prevalence of Bartonella, Borrelia, and Rickettsia species in ticks associated with these wild ruminants. None of the 60 Dermacentor hunteri and 15 Dermacentor albipictus ticks tested yielded positive PCR results. Additional tick specimens should be collected and tested to determine the prevalence of these confirmed or suspected tickborne pathogens within ruminant populations.

Potential disease agents in domestic goats and relevance to bighorn sheep (Ovis canadensis) management.

Domestic goats are raised for meat, milk and hair production, in herds for rangeland weed control, and as pack animals. Domestic sheep, goats and wild bighorn sheep are all susceptible to a multifactorial pneumonia. We sampled 43 herd goats from 7 herds and 48 pack goats from 11 herds for viral and bacterial serology, parasitology, and Pasteurellaceae microbiology. The goats in this study were in generally good health, although most goats did harbor various pathogens and parasites including several bacteria, specifically Pasteurellaceae, which have been associated with pneumonia in free-ranging bighorn sheep. It is not known if domestic goats can transmit the Pasteurellaceae or other pathogens found in this study readily to wild bighorn sheep. However, due the possibility of transmission, domestic goats in areas in or near bighorn sheep habitat should be managed to minimize the risk of spreading disease agents to bighorn sheep.

Reinstatement of Dermacentor kamshadalus Neumann (Acari: Ixodidae) as a valid species parasitizing mountain goats and sheep in the United States, Canada, and Russia.

Reexamination of Dermacentor albipictus (Packard, 1869) holdings stored in the United States National Tick Collection revealed several collections of a morphologically distinct Dermacentor species. Comparison of these specimens with other Dermacentor taxa showed that they are identical to an old taxon originally described as Dermacentor variegatus kamshadalus Neumann, 1908. For more than a century, this taxon was known only from the male holotype specimen collected in Russia, and the name was considered a junior synonym of D. albipictus. D. kamshadalus is reinstated here to a full species rank, and its male is redescribed and its female and nymph are described for the first time. Adults of D. kamshadalus can be distinguished from those of D. albipictus by a short spur on trochanters I, shorter spurs on coxae I, shorter dorsal cornua, more numerous perforations on spiracular plates, less numerous and shorter setae on idiosoma, especially around spiracular plates, and considerably paler coloration of the conscutum and scutum. The nymph of D. kamshadalus can be differentiated from that of D. albipictus by shorter spurs on coxae I and the numerous perforations on the spiracular plates. Adults and nymphs ofD. kamshadalus are recorded from the United States, Canada, and Russia, where they have been collected from mountain goats, Oreamnos americanus (de Blainville), bighorn sheep, Ovis canadensis Shaw, and sheep, Ovis sp. of which the species was not stated.

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.

Muellerius capillaris dominates the lungworm community of bighorn sheep at the National Bison Range, Montana.

Lungworm infections are common among bighorn sheep (Ovis canadensis) in North America, and the predominant species reported are Protostrongylus stilesi and P. rushi. The only records of another lungworm species, Muellerius capillaris, infecting bighorns come from South Dakota, USA. At the National Bison Range (NBR), Montana, USA we found that across six sampling periods, 100% of wild bighorn sheep surveyed were passing first-stage dorsal-spined larvae (DSL) which appeared to be consistent with M. capillaris. By contrast, only 39% or fewer sheep were passing Protostrongylus larvae. Using molecular techniques, we positively identified the DSL from the NBR bighorns as M. capillaris. This is the first definitive record of M. capillaris infection in a free-ranging bighorn sheep population outside of South Dakota.

Experimental infection of bighorn sheep with liver flukes (Fasciola hepatica).

Nine Rocky Mountain bighorn sheep (Ovis canadensis canadensis) were each inoculated orally with 250 metacercariae of Fasciola hepatica. Blood and fecal samples were collected at the time of inoculation and at 5, 10, 14, and 20 wk after inoculation. Numbers of fluke eggs in feces, hemoglobin, packed cell volume, and eosinophil values were determined. Five of the bighorn sheep were treated with triclabendazole at 40 mg/kg of body weight 14 wk after inoculation. Based on fecal evaluations, all bighorns developed patent infections. Six weeks after treatment, fluke eggs were not detected in feces from the five treated animals but were present in two of four untreated animals. One untreated bighorn sheep was euthanized 20 wk after inoculation, and 57 adult F. hepatica were recovered from the liver. Results from this experiment indicated that bighorn sheep are efficient hosts for F. hepatica. Triclabendazole at 40 mg/kg of body weight was safe and, based on fluke egg recovery in feces, apparently an effective treatment. To my knowledge, this is the first published report of F. hepatica in bighorn sheep.

Transmission of lungworms (Muellerius capillaris) from domestic goats to bighorn sheep on common pasture.

Four domestic goats (Capra hircus) that were passing first-stage dorsal-spined larvae of Muellerius capillaris were copastured on a 0.82-ha pasture for 11 mo from May 2003 to April 2004 with seven Rocky Mountain bighorn sheep (Ovis canadensis) that were not passing dorsal-spined larvae. During the 11-mo experiment, two bighorn sheep died from pneumonia caused by Mannheimia (Pasteurella) haemolytica biotype A, serotype 2. The remaining five bighorn sheep and the four domestic goats remained healthy throughout the experiment. Muellerius larvae were detected from all domestic goats on a monthly basis throughout the experiment and were first detected from all five surviving bighorn sheep approximately 5 mo after the copasturing began. Once the bighorn sheep began passing Muellerius larvae, larvae were detected in low numbers from all bighorn sheep every month thereafter for the 6 mo the goats were still in the enclosure and continued to pass larvae for more than 3 yr after the goats were removed from the experiment. Six bighorn sheep in two similar enclosures that did not contain goats did not pass Muellerius larvae before, during, or after the experimental period. Results of this experiment indicate that M. capillaris from domestic goats is capable of infecting bighorn sheep when animals are copastured together on a common range.

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.

Candidate gene microsatellite variation is associated with parasitism in wild bighorn sheep.

The loss of genetic variation in host populations is thought to increase host susceptibility to parasites. However, few data exist to test this hypothesis in natural populations. Bighorn sheep (Ovis canadensis) populations occasionally suffer disease-induced population declines, allowing us to test for the associations between reduced genetic variation and parasitism in this species. Here, we show that individual mean heterozygosity for 15 microsatellite loci is associated with lungworm abundance (Protostrongylus spp.) in a small, recently bottlenecked population of bighorn sheep (linear regression, r2=0.339, p=0.007). This association remains significant for seven microsatellites located in genes (p=0.010), but not for eight neutral microsatellites (p=0.306). Furthermore, heterozygotes at three of four microsatellites located within disease-related genes had lower lungworm burdens. This study corroborates theoretical findings that increased parasitism and disease may be a consequence of reduced heterozygosity in wild populations, and that certain individual loci influence parasite resistance. The results illustrate the usefulness of using genomic information, strong candidate genes and non-invasive sampling for monitoring both genetic variation and fitness-related traits, such as parasite resistance, in natural populations.

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.