Non-invasive assessment of parasitic nematode species diversity in wild Soay sheep using molecular markers |
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Authors: | Wimmer B Craig B H Pilkington J G Pemberton J M |
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Affiliation: | Institute for Animal, Cell and Population Biology, University of Edinburgh, West Mains Road, EH9 3JT Edinburgh, Scotland, UK. b.wimmer@ed.ac.uk |
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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. |
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Keywords: | Parasitic nematodes Parasite diversity Species identification High through-put Soay sheep Natural infection |
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