Application of a sea urchin micronucleus assay to monitoring aquatic pollution: influence of sample osmolality

We have improved our sea urchin micronucleus assay for aquatic samples and used it to evaluate marine pollution. We found that the water samples we had collected for 2 years from the Tokyo bay coast near Tokyo, an industrial megalopolis, were positive due to the water samples being hypo-osmotic rather than to chemical pollutants. The evidence was as follows: (i) the osmolality and salinity of the samples were about half that of sea water; (ii) the micronucleus frequency induced in the water sample decreased to the control level when the osmolality was increased to that of sea water; (iii) artificial sea water diluted with distilled water induced micronuclei dilution-dependently. Since micronucleus induction in the sea urchin assay is influenced by sample osmolality, the osmolality must be adjusted to that of sea water for the assay and osmotic pressure must be considered when evaluating water pollution.