Comparison of molecular and McMaster microscopy techniques to confirm the presence of naturally acquired strongylid nematode infections in sheep

Abstract

Patent strongylid nematode infections were identified using McMaster worm egg counts (WEC) and PCR assays (ITS-2 nuclear ribosomal DNA) to screen genomic DNA extracted directly from lamb faecal samples. Lambs from four different farms in southern Western Australia were sampled rectally on two separate occasions, with McMaster WECs and PCRs conducted on a total of 858 samples. Negative controls (n=96) (WEC<50eggs per gram [epg]) and positive controls (n=96) (faecal samples spiked with a 100μL suspension of third-stage larvae (L3) containing approximately equal proportions of Teladorsagia circumcincta, Trichostrongylus colubriformis, Haemonchus contortus, Oesophagostomum spp. and Chabertia ovina) were generated. All control samples amplified in accordance with positive controls. High levels of agreement (Kappa values≥0.93) were identified between the two diagnostic tests. PCRs detected an additional 2.0% of samples as strongylid-positive but there was no significant difference in the number of strongylid-positive samples identified using PCR or McMaster WEC.