Rapid toxicity assessment using an in vivo enzyme test for Brachionus plicatilis (Rotifera).
A 1-hr in vivo enzyme inhibition assay based on esterase activity has good potential for marine toxicity assessment. A test was developed for the rotifer Brachionus plicatilis based on the nonfluorescent substrate fluorescein diacetate (FDA), which is metabolized by esterases to a fluorescent product. Enzyme inhibition, as determined by reduced fluorescence, can be scored visually or quantified using a fluorometer. Quantification of fluorescence permits the calculation of NOEC, LOEC, chronic value, and IC20. The 1-hr esterase inhibition test has sensitivity comparable to that of 24-hr rotifer acute tests for several compounds. The toxicity of six compounds was examined using the quantified assay. The resulting IC20s were within a factor of 3 of the 24-hour LC50s. IC20 values ranged from 0.017 mg/l for tributyltin to 3.1 mg/l for zinc, with an average coefficient of variation of 17.8%. Electrophoretic analysis of rotifer homogenates suggested that a single C esterase (acetylesterase) was responsible for FDA metabolism in B. plicatilis. Several other aquatic species are capable of metabolizing FDA, including Brachionus calyciflorus, Mysidopsis bahia, Menidia beryllina, Pimephales promelas, Ceriodaphnia dubia, Daphnia pulex, Artemia salina, and Ophryotrocha sp. The esterase inhibition test is an attractive tool for assessing aquatic toxicity because of its speed, simplicity, sensitivity, and applicability to a broad range of aquatic species.
Moffat
B D
BD
Department of Environmental and Occupational Health, University of South Florida, Tampa 33612, USA.
Snell
T W
TW
D000818:Animals; D017610:Calcium Compounds / Q000032:analysis / Q000633:toxicity; D003300:Copper / Q000032:analysis / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D004586:Electrophoresis; D004950:Esterases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D005452:Fluoresceins / Q000378:metabolism*; D005470:Fluorometry; D008628:Mercury / Q000032:analysis / Q000633:toxicity; D010416:Pentachlorophenol / Q000032:analysis / Q000633:toxicity; D012403:Rotifera / Q000187:drug effects* / Q000201:enzymology; D012623:Seawater; D013379:Substrate Specificity; D014220:Trialkyltin Compounds / Q000032:analysis / Q000633:toxicity; D014874:Water Pollutants, Chemical / Q000032:analysis / Q000633:toxicity*; D015032:Zinc / Q000032:analysis / Q000633:toxicity
D017610:Calcium Compounds; D005452:Fluoresceins; D014220:Trialkyltin Compounds; D014874:Water Pollutants, Chemical; C004488:calcium hypochlorite; C011559:tributyltin; D003300:Copper; D010416:Pentachlorophenol; D004950:Esterases; D008628:Mercury; D015032:Zinc; C018506:diacetylfluorescein