The condition of three intertidal Paracentrotus lividus populations, which inhabit an area between Rabat and Casablanca (Morocco) known by its high wave energy and its substantial anthropogenic disturbances were described. The growth rate of the juvenile sea urchins in this intertidal zone was low compared with more sheltered populations. A sharp increase in the gonad index was observed between January and March at the beginning of a period of algal production, and was concomitant with increases in seawater temperature and food consumption rate. This gonad index increase was followed by a single spawning period occurring between March and June, probably triggered by the phytoplankton bloom induced by upwelling conditions. Larval development in all three populations was severely inhibited and could be explained, in part, by high and generalized Cu contamination throughout the region. Specific contamination of particular populations by Pb or Hg was attributed to local industries and the incomplete dispersion of pollutants in spite of the high hydrodynamics along the Moroccan Atlantic coast. One of the three populations studied showed strong indications of abnormal development; the degradation of its condition appeared to be due to the presence of a wadi (temporary river) which can seasonally reduce the salinity, directly affecting the sea urchin physiology and indirectly enhancing the metal toxicity.

Crassostrea gigas D-shaped larvae were subjected to different conditions of temperature and salinity for 24 h and four biomarkers (acetylcholinesterase (AChE) activity, thiobarbituric acid reactive substances (TBARS) levels, glutathione S-transferase (GST) and catalase (CAT) activities) were measured. AChE activity decreased when salinity increased from 25 to 30 and 35 psu at 20 and 25 degrees C. Temperature did not seem to have an influence on AChE activity. TBARS levels increased as a function of salinity when the temperature was maintained at 20 degrees C, whereas at 25 degrees C no effect of salinity could be observed. Variations in GST and CAT activities were not significant with salinity and temperature except that catalase activity was higher at 25 degrees C than at 20 degrees C. Exposure experiments were conducted at 23 degrees C and 30 psu with carbofuran (100 and 1000 microg/l) and malathion (100 and 300 microg/l). There was an inhibition of AChE activity with carbofuran, and a toxic effect shown by an increase in TBARS levels counteracted by increases in GST and CAT activities which protected the larvae. When two pairs of adults producing larvae were taken into consideration, significant differences in biomarker levels were noted between the larval offspring of each pair. Malathion induced a decrease in AChE activity and an increase in CAT activity.

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Genotoxicity biomarkers are widely measured in ecotoxicology as molecular toxic endpoints of major environmental pollutants. However, the long-term consequences of such damage still have to be elucidated. Some authors have suggested that the accumulation of unrepaired DNA lesions could explain the embryotoxicity of certain chemical pollutants. As embryotoxicity exerts a direct impact on the recruitment rate, genotoxicity could be closely related to disturbances of ecological concern and produce a possible impact upon population dynamics. The aim of the present work was to study the genotoxicity and the embryotoxicity of three relevant pollutants for oyster embryos: the polycyclic aromatic hydrocarbon, benzo[a]pyrene (BaP), the synthetic estrogenic hormone, 17alpha-ethinylestradiol (EE2), and the organochlorine pesticide, endosulfan (ES). For each substance, gamete fertilization was performed and embryo development followed in contaminated reference seawater. Following exposure, embryotoxicity was evaluated by calculating the percentage of abnormal D-larvae obtained at 20 h development. Genotoxicity was measured in parallel by conducting a comet assay on enzymatically dissociated cells of pre-shelled larvae (16 h development). The oxidized DNA base, 8-oxodGuo, was also measured by HPLC coupled to electrochemical detection. For each contaminant, the relationship between genotoxicity and embryotoxicity was then studied to check for the possible significance of genotoxicity in the population dynamics of marine bivalves from polluted areas. For BaP, embryotoxicity and DNA strand breakage were both observed from the lowest tested concentration of 0.2 nM. Induction of 8-oxodGuo was significant from 20 nM. Endosulfan exposure resulted in similar effects for oyster embryos but from higher concentrations and followed a concentration-dependent manner. Embryotoxicity and genotoxicity in terms of DNA strand breaks were observed for endosulfan from 300 and 150 nM, respectively. No change in 8-oxodGuo level was observed following endosulfan exposure. EE2 displayed no toxic effect for oyster embryos within the range of tested concentrations (from 0.02 to 1.7 nM). Taking into account all the data collected during this study, a positive and significant correlation was demonstrated in oyster embryos between genotoxicity as measured by the comet assay and embryotoxicity.

Larval stages of bivalve molluscs are highly sensitive to pollutants. Oysters from a hatchery from Normandy (English Channel) were induced to spawn, and fertilized eggs were exposed to copper or cadmium for 24 h. Metal accumulation (from 0.125 to 5 microg Cu L(-1) and from 25 to 200 microg Cd L(-1)) and MT concentrations were measured in larvae. Compared to controls, larvae accumulated copper and cadmium with an increase in MT concentrations particularly with cadmium (i.e. 130.96 ng Cu (mg protein)(-1) and 12.69 microg MT (mg protein)(-1) at 1 microg Cu L(-1) versus 23.19 ng Cu (mg protein)(-1) and 8.92 microg MT (mg protein)(-1) in control larvae; 334.3 ng Cd (mg protein)(-1) and 11.70 microg MT (mg protein)(-1) at 200 microg Cd L(-1) versus 0.87 ng Cd (mg protein)(-1) and 4.60 microg MT (mg protein)(-1) in control larvae). Larvae were also obtained from oysters of a clean area (Arcachon Bay) and a polluted zone (Bidassoa estuary) and exposed to copper in the laboratory, their MT concentration was measured as well as biomarkers of oxidative stress. Biomarker responses and sensitivity to copper for the larvae from Arcachon oysters were higher than for those from Bidassoa.