Measurement of EROD activity: caution on spectral properties of standards used.

The activity of the enzyme 7-ethoxy-resorufin- O-deethylase (EROD) has been extensively used in biomonitoring studies for more than a decade. Although the analytical procedure is simple, it is often poorly characterized. In this study spectral properties of particular standard compounds used to measure EROD activity (ethoxyresorufin and resorufin, standards from Molecular Probes) were tested in order to optimize excitation and emission wavelengths to be used in the fluorimetric assay of EROD activity. The optimal excitation wavelength for the detection of resorufin was 560 nm. At this wavelength the excitation represents only 37% of its maximum level for ethoxyresorufin, while it represents 86% for resorufin. This allows discrimination between the fluorescence emitted by both standards, favoring the formed resorufin. Our results demonstrate that any analytical work using spectrofluorometry to measure EROD activity should be preceded by precise determination of the spectral characteristics of each set of standards used.

Bioaccumulation and toxicity of four dissolved metals in Paracentrotus lividus sea-urchin embryo.

The bioaccumulation of four metals (Cd, Cu, Pb, Zn) dissolved in water was assessed measuring the concentrations recorded within sea-urchin larvae (Paracentrotus lividus, Lmk.) after a 48-h exposure period. Concurrently, the frequencies of abnormalities were evaluated at the 48-h pluteus stage to check the actual toxicity of such contaminants with regards to larval development. Maximum metal concentrations in the larvae reached 43.8 +/- 6.7 micrograms Cd g-1 dry weight (d.w.), 1269 +/- 392 micrograms Cu g-1 d.w., 3106 +/- 679 micrograms Pb g-1 d.w., 534 +/- 62 micrograms Zn g-1 d.w. Bioconcentration factors (BCFs) reached very high values for Pb (maximum value: 16,282 +/- 973) and indicated that the concentrations of Pb and Cd were directly related to the water concentration at least in the range of concentrations tested. Zn appeared to be partially regulated above a certain threshold concentration (between 5 and 25 micrograms l-1). Cu was accumulated with an active uptake at least below lethal threshold level (100 micrograms l-1). Nevertheless, above these thresholds, high mortality of embryos prevent any interpretation of Zn and Cu bioconcentration factors. Concurrent studies carried out on embryotoxicity confirmed the high toxicity of Cu towards sea-urchin pluteus larvae and highlighted the different behaviour of Cd. The internal Cd concentrations were directly related to the external ones, comparable to other non-essential metals, but Cd had little toxic effect on the initial larval development.

Concentration and mobilization of Cd, Cu, Pb and Zn by meiofauna populations living in harbour sediment: their role in the heavy metal flux from sediment to food web.

The bioavailability of Cd, Cu, Pb and Zn was assessed in different harbours of the Atlantic coast (France). The responses of the two dominant meiobenthic groups (nematodes and copepods) to a heavy metal contamination gradient measured in similar subtidal sediments were observed in field surveys. Heavy metal concentrations were measured in nematodes and copepods. Nematodes have higher Cd, Cu, Pb and Zn transfer factors than copepods or other benthic species. The flux of heavy metal through nematodes was estimated and appears to be important compared to plankton in the process of heavy metal transfer to benthic or pelagic food webs.

Contamination of a dredged material disposal site La Rochelle Bay,France.The use of the acetylcholinesterase activity of Mytilus edulis L. as a biomarker of pesticides: the need for a critical approach.

A single aspect of the toxic impact of a dredged material disposal site located near a mussel-farming zone was followed for eight months. Acetylcholinesterase activity (AChE) of Mytilus edulis was investigated as a biomarker for possible contamination by neurotoxic compounds (carbamates and/or organophosphorous pesticides). Our observations showed that the enzymatic activities (including AChE) of these harbour mussels were decreased in sites directly and indirectly influenced (according to hydrodynamic conditions) by the dumping of dredged sediments, suggesting possible contamination by pesticides. The strong correlations observed between AChE activity and growth parameters (length and weight) seems to show, however, that the enzyme activity is also indirectly controlled through growth restriction, which may imply limitation of the development of the nervous system in juveniles. The concentration of total proteins, as well as the spawning process also seem to disturb the assessment of AChE activity. These field observations clearly indicate that the use of this enzyme activity as a biomarker should proceed with caution. For example, the seasonal variability of such activity should be taken into account in a biomonitoring programme.