EHAs were conducted in a microtiter plate format. Briefly, 200 無 of 2% agar was deposited into each well of a 96-well microtiter plate. After this had solidified, 30 無 of nematode egg solution was aliquotted into each well. Water (10 無) was added to the control wells, while 10 無 of cyclotide solution (varying concentrations) was aliquotted into the treatment wells. The final concentration range was 104?833 痢/mL (35?288 然) for EHAs. For each assay, triplicate wells were examined at each concentration. The plates were incubated overnight at 28 蚓 and the unhatched eggs and L1 larvae present in each well were counted. The percentage inhibition of egg hatching was estimated by comparing the percentage hatch at each cyclotide concentration with the percentage hatch in the control wells.