In this study, metal contamination experiments were conducted to investigate the effects of copper and cadmium on the growth of the marine toxic dinoflagellate Alexandrium catenella and on the production of dissolved organic matter (Dissolved Organic Carbon: DOC; Fluorescent Dissolved Organic Matter: FDOM). This species was exposed to increasing concentrations of Cu(2+) (9.93 × 10(-10)-1.00 × 10(-7)M) or Cd(2+) (1.30 × 10(-8)-4.38 × 10(-7)M), to simulate polluted environments. The drastic effects were observed at pCu(2+)=7.96 (Cu(2+): 1.08 × 10(-8)M) and pCd(2+)=7.28 (Cd(2+): 5.19 × 10(-8)M), where cyst formation occurred. Lower levels of Cu(2+) (pCu(2+)>9.00) and Cd(2+) (pCd(2+)>7.28) had no effect on growth. However, when levels of Cu(2+) and Cd(2+) were beyond 10(-7)M, the growth was totally inhibited. The DOC released per cell (DOC/Cell) was different depending on the exposure time and the metal contamination, with higher DOC/Cell values in response to Cu(2+) and Cd(2+), comparatively to the control. Samples were also analyzed by 3D-fluorescence spectroscopy, using the Parallel Factor Analysis (PARAFAC) algorithm to characterize the FDOM. The PARAFAC analytical treatment revealed four components (C1, C2, C3 and C4) that could be associated with two contributions: one, related to the biological activity; the other, linked to the decomposition of organic matter. The C1 component combined a tryptophan peak and a characteristic humic substances response, and the C2 component was considered as a tryptophan protein fluorophore. The C3 and C4 components were associated to marine organic matter production.

We studied seasonal variations in bacterial abundance and succession in phyto- and zooplankton assemblages (particularly small taxa) in two neighbouring shallow bays (near Toulon, Mediterranean Sea, France): Little Bay (polluted, eutrophic), and Niel Bay (less polluted, oligotrophic). In Little Bay, bacteria developed in northern spring and phytoplankton (Dinophyceae > 20 µm) in late northern winter–early spring. Zooplankton levels peaked at the end of northern spring and in autumn; this community was dominated by Oithona nana. In Niel Bay, bacterial levels peaked during northern spring and autumn. Phytoplankton (Dinophyceae, Bacillariophyceae) abundance was low and only peaked in June. Zooplankton levels peaked in northern mid-summer. Little Bay was influenced more by the land and by human activities than by the sea. Seasonal factors (e.g. water temperature) and sudden influences (e.g. rain and, indirectly, Mistral wind) may have modified the succession of the plankton communities in this bay. Successions did not follow Margalef’s model and the classical scheme for zooplankton. Conversely, Niel Bay functioning and plankton assemblages were most influenced by the physical environment of the sea than by the land or by human activities. Successions were closely related to the classical scheme of the Mediterranean Sea.

Dimethylsulfoniopropionate (DMSP) concentrations were observed from October 1999 to September 2000 in a Mediterranean ecosystem (Little Bay of Toulon) submitted to eutrophication. DMSP was measured in the particulate material (DMSP p ), and more specifically in the N90, 5-90 and 0.2-5 Am fractions. DMSP was first converted into dimethylsulfide (DMS) by a cold alkali treatment. DMS was then analysed by gas chromatography equipped with a flame photometric detector (FPD). DMSP p concentrations were relatively high, showing a strong temporal variability with maxima in February-March (58.8 nM). The most elevated values were recorded in the 5-90 Am size class, and represented between 60% and 100% of the total DMSP p . This fraction was mostly composed of Dinoflagellates whose biomass was significantly correlated with DMSP p concentrations. These concentrations showed better correlations with Ceratium furca, Dinophysis acuminata, Prorocentrum arcuatum and also Alexandrium minutum. The intracellular contents of DMSP were much higher in Dinoflagellates (124.9F5.7 mM) than in Diatoms (25.1F1.1 mM). A. minutum produced the strongest intracellular concentrations (3387.6F121.9 mM). High DMSP contents were also found in D. acuminata (477.4F64.3 mM) and P. arcuatum (442.2F22.9 mM). The N90 Am size class had a minor importance in DMSP production, generally below 20% of the total DMSP p . However, DMSP in the N90 Am fraction was well correlated with cladoceran abundance. Plankton cells between 0.2 and 5 Am contained a low part of the DMSP p pool, lower than 20%. Besides, no correlation was found between their algal abundances and DMSP in the 0.2-5 Am size class. Temperature and photoperiod could have influenced the Dinoflagellate development, and consequently, the DMSP p concentrations. The strong autumnal rains affected the composition of the phytoplankton community and the production of sulfur compounds. These particular climatic conditions have induced an increase in nutrient concentrations and a drop in salinity, which may explain the low autumnal DMSP p concentrations.

Bidimensional and monodimensional polyacrylamide gel electrophoresis were used to study protein expression from zooplankton collected in thirteen stations of Toulon Bay (NW Mediterranean). In this ecosystem, Little Bay showed higher trace metal concentrations (13.5-23.8 nM for Cu, 0.73-1.24 nM for Pb, 27.8-58.7 nM for Zn) than Large Bay (Cu 2.2-15.6 nM; Pb 0.19-0.78 nM; Zn 9.0-38.8 nM). Trace metals positively correlated (p < 0.05) with expression of four zooplankton proteins (MW in kDa/pI: 25.0/5.6; 48.8/4.1; 38.2/4.4; 38.3/5.8) and with biomass of Oithona nana, predominant copepod in Little Bay. Sequencing by LC-MS/MS putatively provided zooplankton identity of these proteins: they were cytoskeleton actin, except one protein that was the chaperone calreticulin. We suggest that actin and calreticulin could be regarded as zooplankton markers of metal stress and be involved in a possible tolerance of O. nana to contamination, contributing to its development in a marine perturbed ecosystem.

A Pseudomonas fluorescens strain was isolated from oxic marine sediments obtained from the strand zone of the St Anne Bay (a moderately metal-contaminated site to the west of Cherbourg harbour). The strain, which exhibited a high tolerance to metal contamination when cultivated (minimal inhibitory concentration=950 microM [62 mg L(-1)] for Zn, 660 microM [42 mg L(-1)] for Cu, and 505 microM [57 mg L(-1)] for Cd), was further characterized by its physiological and biochemical responses to metal additions to the culture medium. Bacterial growth was significantly disturbed by 380 microM Zn (25 mg L(-1)), 315 microM Cu (20 mg L(-1)) and 90 microM Cd (10 mg L(-1)). The Zn-containing alkaline phosphatase was studied as an intoxication biomarker. Its activity was stimulated (+9%) by an excess of Zn, but inhibited by Cd (-55%) and Cu (-10%), these two elements could displace the native Zn or/and disturb the enzyme 3D-structure. Bacterial O(2) consumption was recorded as a global physiological response to metal stress. This parameter dropped with increasing Cd and Cu contamination (-49% and -45%, respectively, at 20 mg L(-1)). By contrast, Zn increased O2 consumption (approximately +40% for the different tested concentrations). The proteomes of bacteria grown in the presence or absence of 20 mg metal L(-1) were characterized by 2D-gel electrophoresis. The number of spots exhibiting a difference in intensity between the contaminated sample and the control was 65, 68, and 103, for Zn, Cu and Cd, respectively. Among them, 45, 61 and 82 spots respectively appeared de novo or increased in intensity, indicative of metal-stimulated synthesis, particularly for Cu and Cd. In summary, whereas Cd and Cu treatments both stressed cells and slowed down primary metabolism to differing extents, Zn has a stimulating action on several physiological and biochemical parameters.