The experiment followed a factorial design, and
combinations were made from five temperatures (15,
20, 25, 30 and 35 ºC) and four salinities (15, 25, 35
and 45 ‰). In each combination 3 random females
were used from a minimum set of 6 hatchings, each
female with 2 replicates of 15 larvae. Larvae were
chosen from amongst the most active of respective
batches. Each replicate was cultured in 400 ml
glass bowls. Water was periodically brought from
the shore, and filtered through a series of Millipore
filters down to 0.5 µm. Dilution was done using
de-ionised water and concentration was obtained
by evaporation. Temperatures were regulated in
enclosed chambers to precision of within ± 0.5 ºC.
Larvae were transferred to new cultures each
day and fed on newly-hatched Artemia nauplii.
While transferring, larvae were checked for
mortality and sorted by developmental stage. At
high temperatures (30 and 35 ºC) the cultures were
covered with adhesive plastic to prevent
evaporation and consequent salinity increase. The
experiment lasted until the juvenile stage was
reached.