Molecular identification of naturally acquired strongylid infections in lambs--an investigation into how lamb age influences diagnostic sensitivity

Faecal samples (n=1155) were collected from n=111 (Farm A) and n=124 (Farm B) 2-6 week old female lambs on two farms in southern Western Australia across five sampling occasions (spanning 8 months). Genomic DNA was extracted directly from faecal samples and screened by PCR for ITS-2 nuclear ribosomal DNA to detect patent strongylid infections, specifically Teladorsagia circumcincta, Trichostrongylus spp., Haemonchus contortus, Oesophagostomum spp. and Chabertia ovina. The minimum amount of extracted genomic DNA necessary for successful PCR amplification was 2.0-5.0 pg. During the five sampling occasions for the two farms, the sensitivities for WEC and PCR identification of strongylid infections varied, with levels of agreement between the two sets of diagnostic results ranging from 85 to 100%. Strongylid species prevalences were high (90.3-97.3%), with T. circumcincta and Trichostrongylus spp. the most prevalent species and together they were the most common mixed strongylid infection; H. contortus was not identified in either flock. T. circumcincta was the only species associated with an increased risk of non-pelleted faeces on Farm B, where T. circumcincta-positive lambs were 2.3 and 2.6 times more likely to have non-pelleted faeces than negative lambs at the second and final samplings, respectively. The highest strongylid prevalence, mixed strongylid prevalence and mean number of strongylid species detected per lamb coincided with the highest average flock faecal worm egg counts (WECs) on both farms. There was a positive correlation between the number of strongyle species detected per lamb and both WEC and adjusted WEC (P<0.01; r(2) 0.026-0.591). These results indicate that strongylid eggs were likely to be the main source of strongylid DNA in the faecal DNA extracts. Despite the progress made by the molecular approach utilised in this study, it is incapable of distinguishing between patent and non-patent sources of strongylid DNA. However there is potential for further investigation into the development of a similar molecular procedure which could be used for early larvae detection on pastures.