Preparation of Animals for Test: - Remove all young daphnia culture bottles with a

pipette 6 hours prior to the beginning of the test. Wash daphnia three diluent water by
transferring to a bottle containing the diluent water and allow them to remain in water
for 5 minutes.
Procedure
Preliminary Test: - Prepare dilution covering a wide range of concentrations for
example 100, 10, 1, 0.1 present with diluted water. Place 10 or more of the daphnia in
each concentration and observed for a period of 24 hours.
Full Scale Bio-Assay
Set up six 150 ml bottles the various concentration of the sample under investigation
using diluted waters. Concentration in a series of dilution using logarithmic valued are
preferable. Keep the sixth bottle with dilute water only as control. Transfer 50 daphnia
prepared for test in each bottle. Observe and record the number of daphnia which are
imm obalised in each their container at the end of 12 hours and 24 hours, ater their
introduction and also at the end of 48 hours, if the test are continued beyond 24 hours.
Remove all young daphnia culture bottles with a pipette 6 hours prior to the beginning
of the test. Wash daphnia three diluent water by transferring to a bottle containing the
diluent water and allow them to remain in water for 5 minutes.
covering a wide range of concentrations for example 100, 10, 1, 0.1 present with diluted
water. Place 10 or more of the daphnia in each concentration and observed for a period
of 24 hours.
150 ml bottles the various concentration of the sample under investigation using diluted
waters. Concentration in a series of dilution using logarithmic valued are preferable.
Keep the sixth bottle with dilute water only as control. Transfer 50 daphnia prepared for
test in each bottle. Observe and record the number of daphnia which are imm obalised
in each their container at the end of 12 hours and 24 hours, ater their introduction and
also at the end of 48 hours, if the test are continued beyond 24 hours.