title: Non-invasive assessment of parasitic nematode species diversity in wild Soay sheep using molecular markers.
abstract: 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.
authors.affiliation: Institute for Animal, Cell and Population Biology, University of Edinburgh, West Mains Road, EH9 3JT Edinburgh, Scotland, UK. b.wimmer@ed.ac.uk
authors.forename: B
authors.initials: B
authors.lastname: Wimmer
authors.forename: B H
authors.initials: BH
authors.lastname: Craig
authors.forename: J G
authors.initials: JG
authors.lastname: Pilkington
authors.forename: J M
authors.initials: JM
authors.lastname: Pemberton
mesh_terms: D000723:Ancylostomatoidea / Q000302:isolation & purification; D000818:Animals; D001483:Base Sequence; D044822:Biodiversity; D015415:Biomarkers / Q000032:analysis; D018509:DNA, Helminth / Q000032:analysis; D021903:DNA, Ribosomal Spacer / Q000032:analysis; D004021:Dictyocaulus / Q000302:isolation & purification; D005243:Feces / Q000469:parasitology; D008969:Molecular Sequence Data; D009348:Nematoda / Q000302:isolation & purification*; D009349:Nematode Infections / Q000469:parasitology / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000379:methods; D012756:Sheep; D012757:Sheep Diseases / Q000469:parasitology*; D013045:Species Specificity; D014254:Trichostrongylus / Q000302:isolation & purification; D014258:Trichuris / Q000302:isolation & purification
chemical_list: D015415:Biomarkers; D018509:DNA, Helminth; D021903:DNA, Ribosomal Spacer