Figure 4
Activation and inhibition of H. contortus L3 to L4 development in vitro. (a) Δ7-Dafachronic acid (Δ7-DA) was added to EBSS culture medium/5% CO2/37 °C at 1–10 μM final concentration and development from L3 to L4 stage examined microscopically. Control wells contained the same volume of ethanol (DA solvent). Percentage of H. contortus L3 and L4 larvae after 72 h in the presence or absence of DA is shown. Significant differences were observed at all DA concentrations (two tailed t-test P = 0.0039 to P = 0.000056). Mean and standard deviation of three experiments are shown. (b) Morphology of L3 and L4 stages in vitro. Dark gut granules (arrow) and closed buccal cavity (asterisk) are observed in L3 larvae, while L4 stage show an open gut lumen (arrowhead) and open buccal cavity (asterisk). (c) Percentage of H. contortus L3 and L4 larvae after five days in the presence of inhibitors of both hco-mir-228 and hco-mir-235 (final concentration 2 μM). Inhibitors enhanced development from L3 to L4 relative to control inhibitor at same final concentration (t-test *P = 0.038). Mean and standard deviation of three replicate experiments are shown.
