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MERC+GSI – DRAFT Test Plan
32
Table 8. Summary of Test Conditions and Test Acceptability Criteria for the Algae
Isochrysis galbana
and
Tetraselmiss uecica
Chronic Growth Test
Test type:
Static non-renewal (required)
Temperature:
20 ºC ± 1 ºC
Light quality
“Cool white” fluorescent lighting
(recommended)
Light intensity:
360-440 foot candles
Photoperiod:
Continuous illumination
Test chamber size:
250 ml
Test solution volume:
100 ml
No. replicate chambers per concentration:
4
Renewal of test solutions:
None
Age of test organisms:
Log growth phase
Initial cell density in test chambers:
1-2 X 10
4
cells/ml
Shaking rate:
100 rpm continuous on a mechanical shaker or
twice a day hand shaken
Aeration:
None
Nutrient solution
Algal assay culture medium nutrients added to
each replicate (Appendix A3 of ASTM
Designation E 1218-04; ASTM, 2006)
Dilution water:
Baltimore Harbor water collected at the same
time as the initial untreated ballast water
Test concentrations:
100, 56, 32, 18, and 10 % ballast discharge or
receiving water by volume plus a Baltimore
Harbor and a Wye River control
Dilution factor:
0.56 dilution series
Test duration:
96 hours
Endpoint:
Growth (cell counts)
Test acceptability criterion:
Mean cell density of at least 1 x 10
6
cells/ml in
the controls; and variability (CV%) among
control replicates less than or equal to 20%