New host and geographic records for two protostrongylids in Dall's sheep.

Biodiversity survey and inventory have resulted in new information on the distribution of Protostrongylidae in Dall's sheep (Ovis dalli dalli) from the Northwest Territories (NT, Canada) and from Alaska (AK, USA). In 1998, Parelaphostrongylus odocoilei adults were found for the first time in the skeletal muscles of Dall's sheep in the Mackenzie Mountains (NT). Adult P. odocoilei were associated with petechial and ecchymotic hemorrhages and localized myositis; eggs and larvae in the lungs were associated with diffuse granulomatous pneumonia. Experimental infections of the slugs Deroceras laeve and Deroceras reticulatum with dorsal-spined first-stage larvae assumed to be P. odocoilei, from ground-collected feces from Dall's sheep in the Mackenzie Mountains, yielded third-stage larvae by at least 28 (in D. laeve) and 48 (in D. reticulatum) days post-infection. Third-stage larvae emerged from D. laeve between days 19 and 46 post-infection and emergence occurred both at room temperature and at 10 to 12 C. Protostrongylus stilesi were definitively identified from the lungs of Dall's sheep collected in the Mackenzie Mountains, NT in 1998. Specimens collected from sheep in the Mackenzie Mountains, NT in 1971-72, and the Alaska Range, AK in 1972 were also confirmed as P. stilesi. Lung pathology associated with adults, eggs, and larvae of P. stilesi was similar to that described in bighorn sheep (Ovis canadensis). Concurrent infections with P. odocoilei and P. stilesi in a single host have not been previously reported.     

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.     

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.     

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.     

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.     

Parelaphostrongylus odocoilei in Columbian black-tailed deer from Oregon

Documenting the occurrence of Parelaphostrongylus odocoilei has historically relied on the morphological examination of adult worms collected from the skeletal muscle of definitive hosts, including deer. Recent advances in the knowledge of protostrongylid genetic sequences now permit larvae to be identified. Dorsal-spined larvae (DSLs) collected in 2003-2004 from the lung and feces of six Columbian black-tailed deer (Odocoileus hemionus columbianus) from Oregon were characterized genetically. The sequences from unknown DSLs were compared to those from morphologically validated adults and larvae of P. odocoilei at both the second internal transcribed spacer (ITS-2) of ribosomal DNA and the mitochondrial cytochrome oxidase II gene. We provide the first unequivocal identification of P. odocoilei in Columbian black-tailed deer from Oregon. The broader geographic distribution, prevalence, and pathology of P. odocoilei are not known in populations of Oregon deer.     

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.     

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.     

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.     

Patterns in size and shedding of Fasciola hepatica eggs by naturally and experimentally infected murid rodents

Using samples collected on the island of Corsica, a comparative study was done of the morphometry of Fasciola hepatica eggs shed by cattle and by naturally and experimentally infected murid rodents (wild Mus musculus and Rattus rattus and Rattus norvegicus Wistar laboratory strain). Eggs shed by murids are smaller in size than those shed by naturally infected cattle. A second study analyzed the number of F. hepatica eggs shed in murid feces at different time intervals, i.e., months, days, and 6-hr periods, by the Kato-Katz technique. Both experimentally and naturally infected black rats (R. rattus) were used, and Wistar rats were experimentally infected and included for comparison. The present studies prove that black rats R. rattus are able to shed eggs independently from the liver fluke isolate and that egg shedding occurs throughout the life of this host species, uninterrupted during all the months analyzed in a 2-yr period. Moreover, the results suggest that this shedding is continuous, with eggs appearing in the feces daily. The results on egg shedding by wild black rats R. rattus reach their maximum shedding in spring and autumn and a maximum during twilight hr. These chronobiological patterns appear to favor parasite transmission, both seasonally and daily.     

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.     

Frequency of deposition and location of Baylisascaris procyonis eggs in raccoon feces

Baylisascaris procyonis is a large ascarid nematode found in the small intestine of raccoons (Procyon lotor). Infection with larvae of B. procyonis can produce visceral, ocular, and neural larval migrans in humans. Infected raccoons can shed millions of eggs a day in their feces. However, it is unknown whether eggs are consistently shed or whether eggs occur at irregular intervals by the population of female nematodes within a host. We trapped, infected, and collected daily fecal samples from 11 raccoons maintained in captivity. Eggs from B. procyonis were obtained from anterior, central, and posterior sections of raccoon feces, isolated by flotation, and quantified under 100× magnification. Naturally infected raccoons were collected and used as a comparison with the experimentally infected group. All raccoons in the experimental group (n=11) became infected with B. procyonis after consuming one infected mouse. Additionally, differential egg deposition rates were observed among individual raccoons from the experimental and naturally infected groups. Mean number of eggs per gram of feces (means±SE) was 16,563±4,321, which was less than previously reported for the species. However, no differences (F(2,30)=0.84, P=0.45) were noted in mean number of eggs per gram of feces among fecal sections. Wildlife biologists, veterinarians, health officials, and researchers of B. procyonis should collect daily fecal samples for a minimum of 3 days before identifying a raccoon as negative for B. procyonis infection. However, it does not matter where within the fecal matter the sample is obtained.     

Parasites of the Dall's porpoise (Phocoenoides dalli True)

The prevalences of three helminths, Campula oblonga, Halocercus dalli and Crassicauda sp., recovered from Dall's porpoises which were net-entrapped incidentally in the vicinity of the Western Aleutian Islands in the northwest Pacific are reported. Specimens of Campula oblonga were found within the bile ducts of 46% of 127 livers examined. The prevalence of hepatic trematodiasis increased with the age of the host. Pulmonary nodules associated with Halocercus dalli were noted in 71% of the Dall's porpoises. Adult H. dalli were recovered from the main stem bronchi of heavily infected lungs. Younger animals exhibited a relatively higher prevalence. Specimens of Crassicauda sp. were found within the main lactiferous canal of 69% of 29 mammary glands examined. The prevalence was highest in mature porpoises. Possible detrimental effects and the modes of transmission of the three species of parasites are also considered.     

The potential for false-positive diagnosis of protostrongyliasis by extraction of larvae from feces

The potential of protostrongylid first-stage larvae (L1) to survive passage through the alimentary canal of non-infected mammals was investigated. Parelaphostrongylus tenuis L1 were collected from feces of an experimentally infected white-tailed deer (Odocoileus virginianus). We utilized two red deer (Cervus elaphus) and four laboratory rats (Rattus norvegicus) which were each fed the L1 of P. tenuis. Larvae were recovered, intact and alive, from the fecal samples of all six animals. Larvae of P. tenuis, and probably of other related species, can survive passage through the alimentary canal of uninfected mammals and they can be collected from feces using the Baermann technique and other related larval extraction methods. Rain water was found to be successful in the dispersal of P. tenuis L1 from the feces of infected animals. These findings raise the possibility of ingestion of L1 and their subsequent passage, by uninfected animals. This potential for false-positive diagnosis of infection in live animals necessitates accurate interpretation of a host's infection-status. Such findings reinforce the need for a reliable method of diagnosing infections in live animals.     

Identification of dorsal-spined larvae from free-ranging wapiti (Cervus elaphus) in southwestern Manitoba, Canada

Dorsal-spined first-stage larvae recovered from feces of free-ranging wapiti (Cervus elaphus) were passaged through snails (Triodopsis multilineata) and two hand-raised white-tailed deer fawns (Odocoileus virginianus). A total of 74 adult Parelaphostrongylus tenuis were recovered from the fawns; no other protostrongylid nematodes were recovered. The study indicates that wapiti may be infected with natural infections of meningeal worm and pass larvae suitable for transmission to gastropod intermediate hosts. Wapiti from areas endemic with P. tenuis should not be translocated to areas currently free of the parasite.     

An ultrasensitive capture ELISA for detection of Fasciola hepatica coproantigens in sheep and cattle using a new monoclonal antibody (MM3)

A capture enzyme-linked immunosorbent assay (ELISA) using a new monoclonal antibody (mAb MM3) is reported for the detection of Fasciola hepatica excretory-secretory antigens (ESAs) in feces of infected hosts. The mAb MM3 was produced by immunization of mice with a 7- to 40-kDa purified and O-deglycosylated fraction of F. hepatica ESAs, which has previously been shown to be specific for the parasite. The specificity and sensitivity of the MM3 capture ELISA were assessed using feces from sheep and cattle. Sheep feces were obtained from a fluke-free herd (with most animals harboring other nematodes and cestodes), from lambs experimentally infected with 5-40 F. hepatica metacercariae and in some cases treated with triclabendazole at 14 wk postinfection (PI), and from uninfected control lambs. Cattle feces were collected at the slaughterhouse from adult cows naturally infected with known numbers of flukes (from 1 to 154) or free of F. hepatica infection (though in most cases harboring other helminths). The MM3 capture ELISA assay had detection limits of 0.3 (sheep) and 0.6 (cattle) ng of F. hepatica ESA per milliliter of fecal supernatant. The assay detected 100% of sheep with 1 fluke, 100% of cattle with 2 flukes, and 2 of 7 cattle with 1 fluke. The false-negative animals (5/7) were probably not detected because the F. hepatica individuals in these animals were immature (5-11 mm in length). As expected, coproantigen concentration correlated positively (r = 0.889; P < 0.001) with parasite burden and negatively (r = 0.712; P < 0.01) with the time after infection at which coproantigen was first detected. Nevertheless, even in animals with low fluke burdens (1-36 parasites), the first detection of F. hepatica-specific coproantigens by the MM3 capture ELISA preceded the first detection in egg count by 1-5 wk. In all sheep that were experimentally infected and then untreated, coproantigen remained detectable until at least 18 wk PI, whereas in sheep that were experimentally infected and then flukicide treated, coproantigen became undetectable from 1 to 3 wk after treatment. None of the fecal samples from sheep or cattle negative for fascioliasis but naturally infected with other parasites including Dicroelium dendriticum showed reactivity in the MM3 capture ELISA. These results indicate that this assay is a reliable and ultrasensitive method for detecting subnanogram amounts of F. hepatica antigens in feces from sheep and cattle, facilitating early diagnosis.     

Non-invasive assessment of parasitic nematode species diversity in wild Soay sheep using molecular markers

Considerable effort has been put into detecting and identifying parasitic nematodes in live ruminants, but to date most studies are limited to a small group of nematodes and/or to experimentally infected sheep. In this study, a PCR-based assay using species-specific primer pairs, located in the second internal transcribed spacer ribosomal DNA, was developed to identify nine different species from six different families of parasitic nematodes in a wild, unmanaged and naturally infected population of sheep. Each primer pair was tested for its specificity and sensitivity and it exclusively amplified the species it was designed for and exhibited a high degree of sensitivity. The method was applied to eggs and cultured larvae to identify the parasitic nematodes present in a pooled faecal sample from several host individuals with unknown parasite burden. To test detection reliability, a faecal sample from an individual with known parasite burden (through post-mortem analysis) was also examined. All species present could be correctly identified by PCR, but detecting very low levels and/or early stages of infection proved to be difficult. The method was also tested for its applicability to high through-put screening of faecal samples.