1. Test type: Static, renewal 2. Temperature (0C). 25 + 1C 3. Light qua1ity. Ambient 1aboratory illumination 4. Photoperiod: 16 hr. light, 8 hr. dark 5. Test chamber size: 30 mi 6. Test solution vo1ume: 15 mi 7. Renewa1 of test solutions: Daily using most recently collected sample 8. Age of test organisms: Less than 24 hr.; and all released within an 8 hr. period of each other. 9. Number of' neonates per test chamber: 1 10. Number of replicate test chambers per treatment: 10 11. Number of neonates per test concentration: 10 12. Feeding regime: Feed 0.1 ml each of YTC and algal suspension per exposure chamber daily. 13. Aeration: None 14. Dilution water: Receiving Water, other surface water, moderately hard synthetic water (prepared using either Millipore MilliQ® or equivalent deionized water and reagent grade chemicals) or deionized water combined with mineral water. 15. Effluent concentrations:³ 5 concentrations and a control. An additiona1 dilution at the permitted effluent concentration (% effluent) is required if it is not included in the dilution series. 16. Dilution factor: 0.5 17. Test duration: Until 60% of control females have three broods (may require 7 days). 18. End Points: survival and reproduction 19. Test acceptability: 80% or greater survival and an average of 15 or more young/fema1e in the control so1utions. At least 60% of surviving females in controls must have produced third brood. 20. Sampling requirements: For on-site tests, samples are collected daily and used within ~4 hr. of the time they are removed from the samp1ing device. For offsite tests, a minimum of three samp1es are collected (e.g. days 0, 3, 5) and used for renewal (see Sec IV). Off-site tests samples must be first used within 48 hours of collection
