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Sugars were extracted with four successive 250 l aliquots of 80% (v/v) ice-cold ethanol and further grinding.
Each pool was divided in two equal aliquots and processed in duplicate.
Due to the small amount of material available, only 15 aliquots of the sample could be prepared by settling in acetone on aluminum discs.
Two 100 ml aliquots were taken from the vials and the number of nematodes contained in 100 ml was determined under stereo-microscope.
If helminth eggs were found in any of the five aliquots of a given sample, this was recorded as 'contaminated'.
Further serial dilutions of the cercarial suspension were made to give five aliquots having densities of 20, 15, 10, 5 and 2.5 cercariae ml21.
A suspension of eggs was fertilised at time 0 and aliquots fixed at the indicated time points.
The drug solutions were prepared from frozen aliquots of stock solution, dissolved in extracellular solution, and applied from seven-barrel pneumatic puffer pipettes.
The supernatants were divided in two aliquots for further analysis.
At the start and after 5 h of incubation, aliquots were removed and analysed.
The remaining homogenate was centrifuged for 2 min at 10,000 g before aliquots of supernatant were removed for the following assays.
Positive results were confirmed by repeating the assay using two aliquots from the same sample.
The bags were checked daily for evidence of green colour and aliquots were removed for absorbance spectra after 8 days.
Two percent aliquots of the total worm burden were counted.
The total number of eggs was calculated by counting eggs in multiple representative aliquots of each collection.
