Sea-urchin Embryo Bioassay for in situ Evaluation of the Biological Quality of Coastal Seawater

The Paracentrotus lividus sea-urchin embryo bioassay, consisting of incubation of fertilized eggs in test water and measurement of the percentage of four-armed plutei larvae developed after the incubation period (2–3 days), has been adapted for in situ evaluation of seawater quality in coastal areas. Mature sea-urchins are dissectedin situ and fertilization is performed in the field; fertilized eggs are delivered into screw lid 50-ml cylinders with 20μm nylon mesh in both ends filled with sieved local seawater. The cylinders, tied to 60-cm ropes with weights on one end and buoys in the other one, are placed by scuba divers in the test sites at subtidal level and recovered after the incubation period. The contents of each cylinder are then transferred into a vial, fixed with formalin and observed directly under an inverted microscope to record the percentage (N=100) and size (length, N=25) of four-arm pluteus larvae. Our results show that the bioassay can discriminate between well known polluted and unpolluted sites, but further improvement is needed in order to: (1) take into account differences of temperature between sites; (2) minimize larval mortality due to reasons other than pollution.