For each analysis, approximately 200,000 nauplii were collected on 50 mm diameter
discs of 44 mm nylon screen. Fatty acids were analysed using techniques modified from
Dunstan et al. Ž1992.. Animals were rinsed into a reaction tube with dichloromethane
ŽDCM.:methanol:water Ž15 ml. and the sample homogenised and stored under nitrogen
at 5–68C for 12 h. Samples were filtered under gentle vacuum and rinsed with
DCM:methanol:water Ž50 ml.. Lipids were extracted in DCM from a mixture of
DCM:water Ž1:1 vrv.. The solvent was removed using a Bucciw rotary evaporator and
the remaining lipid dissolved in methanol Ž5 ml.. Samples were transferred to 20 ml
reaction tubes, combined with acidified methanol Ž4 ml., toluene Ž2 ml. and an internal
standard Žnonadecaenoic acid; 0.5 mg. and heated to 508C for 16 h. The cooled reaction
mixture was transferred to hexane Ž20 ml., washed twice with deionised water Ž20 ml.
and dried over anhydrous sodium sulphate.