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.
Wimmer
B
B
Institute for Animal, Cell and Population Biology, University of Edinburgh, West Mains Road, EH9 3JT Edinburgh, Scotland, UK. b.wimmer@ed.ac.uk
Craig
B H
BH
Pilkington
J G
JG
Pemberton
J M
JM
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
D015415:Biomarkers; D018509:DNA, Helminth; D021903:DNA, Ribosomal Spacer