SYMPOSIA PAPER Published: 01 January 1985
STP33589S

Rapid Aquatic Toxicity Assay Using Incorporation of Tritiated-Thymidine into Sea Urchin, , Embryo: Evaluation of Toxicant Exposure Procedures

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Toxicity of substances in seawater was measured using growth inhibition of embryonic sea urchins during a short period after fertilization. Growth of Arbacia punctulata embryos was monitored by incorporation of tritium-labeled thymidine, a nucleoside incorporated into newly synthesized deoxyribonucleic acid (DNA). Sensitivity and ranking ability of this rapid Arbacia embryo test were judged by comparison of toxicity values for several organic industrial chemicals using this test and two standard acute aquatic toxicity tests. Median effective toxicity values (EC50 values) for early embryos compared favorably to median lethal values (LC50 values) obtained in standard acute toxicity tests using Pimephales promelas (fathead minnow) and Daphnia magna (water flea).

This paper presents a comparison of toxicant exposure procedures using the Arbacia embryo thymidine incorporation test. Toxicant exposure began before, at the time of, or after fertilization and continued for 4 h following fertilization. In addition to the eight organic chemicals tested for comparison to acute toxicity values for other species, several chemicals with embryotoxic potentials (tumor promoters and teratogens) were tested to determine differential sensitivities of exposed life-stages: unfertilized egg, fertilization, and early embryo. EC50 values for any one substance were not significantly changed by exposure modification. Toxicity values for exposures that included fertilization as well as early embryo growth were at least as sensitive as post-fertilization exposure values for all compounds tested except one. Because of technical ease and potential sensitivity, toxicant exposure that includes fertilization as well as early embryo growth (but not unfertilized egg exposure) is recommended for future testing.

Additional development, refinement, and standardization of this fertilization and early embryo test should further increase its utility in toxicity testing. Because this test requires only 4 h to exposure and can be correlated with standard measurements of toxicity, it could be used in early stage hazard evaluation in conjunction with other aquatic bioassays.

Author Information

Nacci, DE
JRB Associates, Marine Services Branch, Narragansett, RI
Jackim, E
U.S. Environmental Protection Agency, Narragansett, RI
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Developed by Committee: E47
Pages: 382–394
DOI: 10.1520/STP33589S
ISBN-EB: 978-0-8031-4952-6
ISBN-13: 978-0-8031-0437-2