Improved bioassay for estimation of inhibitory effects of ovine gastrointestinal mucus and anthelmintics on nematode larval migration

Abstract

A bioassay based on the ability of substances present in ovine gastrointestinal mucus or anthelmintics to paralyse third stage larval (L3) nematodes and inhibit their passage through 20 μm nylon mesh sieves [larval migration inhibition (LMI) activity] is described. Factors influencing the reproducibility of the bioassay were examined using exsheathed L3 of the sheep gastrointestinal nematode parasite Trichostrongylus colubriformis. Levamisole, morantel tartrate and piperazine were shown to inhibit L3 migration in the bioassay. The bioassay was used to demonstrate that gastrointestinal mucus from nematode-resistant sheep possessed greater L3 inhibitory activity than mucus from nematode-susceptible sheep. LMI activity of mucus used in this bioassay was significantly correlated with LMI activities obtained using two previously described similar bioassays. The action of mucus components on nematode larvae was shown to be reversible in the bioassay. The modified assay has advantages over other bioassays as it avoided the use of temperature-dependent agar blocks, reduced the number of L3 required to a more manageable size, and the whole experiment could be performed on a 48-well culture plate. The reproducibility, high correlation with other bioassays, ease of performance, suitability for testing a large number of samples and low cost make this modified assay the method of choice for determining antiparasitic activity of gastrointestinal mucus components and as a screen for potential anthelmintics.