The temporal variability of the abundance and the incorporation of (3)H-thymidine and (14)C-glucose by attached and free-living bacteria, as well as their relation with environmental factors, were analyzed in a coastal marine ecosystem during a year. Both communities were quantitatively very different. Attached bacteria represented only 6.8% of the total bacterial abundance, whereas free-living bacteria represented 93.2%. The environmental factors most closely linked to the abundance and activity of free-living bacteria were temperature and the concentration of dissolved nutrients. Moreover, the free-living community showed similar temporal variations in abundance and in activity, with lower values in the cold months (from October to May). The attached community did not present the same pattern of variation as the free-living one. The abundance of the attached bacteria was mainly correlated to the concentration of particulate material, whereas their activity was correlated to temperature. We did not find a significant correlation between the abundance and the activity of the attached community. On the other hand, the activity per cell of the two communities did not present a clear temporal variation. Attached bacteria were more active than free-living ones in the incorporation of radiolabeled substrates on a per cell basis (five times more in the case of glucose incorporation and twice as active in thymidine incorporation). However, both communities showed similar specific growth rates. The results suggest that the two aquatic bacterial communities must not be considered as being independent of each other. There appears to be a dynamic equilibrium between the two communities, regulated by the concentrations of particulate matter and nutrients and by other environmental factors.

The kinetics of bacterial hydrolytic ectoenzymatic activity and the uptake of monomeric compounds were investigated in the Northwestern Mediterranean Sea. Aminopeptidase and alpha- and beta-glucosidase activities were analyzed by using fluorogenic substrates at 15-22 concentrations ranging from 1 nM to 500 µM. Radiolabeled glucose and a mixture of amino acids were chosen as representatives of monomeric compounds, and the bacterial uptake rates (assimilation plus respiration) were determined over a wide range of substrate concentrations (from 0.2 nM to 3 µM). We found biphasic kinetics both for hydrolytic enzymes and uptake systems: high affinity enzymes at low concentrations of substrates (Km values ranged from 48 nM to 2.7 µM for ectoenzymes and from 1.4 nM to 42 nM for uptake systems), and low affinity enzymes at high concentrations of substrates (Km values ranged from 18 µM to 142 µM for ectoenzymes and from 0.1 µM to 1.3 µM for uptake systems). Transition between high and low affinity enzymes was observed at 10 µM for aminopeptidase and from 1 µM to 25 µM for glucosidases, and it was more variable and less pronounced for the uptake of glucose (40 nM-0.28 µM) and amino acids (10 nM-0.16 µM). Results showed that the potential rates of hydrolysis and uptake are tightly coupled only if the high affinity hydrolytic ectoenzymes and the low affinity uptake systems are operating simultaneously.

Production and specific growth rates of attached and free-living bacteria were estimated in an oligotrophic marine system, La Salvaje Beach, Vizcaya, Spain, and in a freshwater system having a higher nutrient concentration, Butron River, Vizcaya, Spain. Production was calculated from [methyl-3H]thymidine incorporation by estimating specific conversion factors (cells or micrograms of C produced per mole of thymidine incorporated) for attached and free-living bacteria, respectively, in each system. Conversion factors were not statistically different between attached and free-living bacteria: 6.812 x 101" and 8.678 x 1011 ,ug of C mol-' for free-living and attached bacteria in the freshwater system, and 1.276 x 10" and 1.354 x 101" ,ug of C moli-for free-living and attached bacteria in the marine system. Therefore, use of a unique conversion factor for the mixed bacterial population is well founded. However, conversion factors were higher in the freshwater system than in the marine system. This could be due to the different trophic conditions of the two systems. Free-living bacteria contributed the most to production in the two systems (85 % in the marine system and 67 % in the freshwater system) because of their greater contribution to total biomass. Specific growth rates calculated from production data and biomass data were similar for attached and free-living bacteria.

Bacterivory on aggregates and bulk seawater by three nanoflagellates and a microciliate was investigated. A new approach to measure bacterivory on the attached bacterial community is proposed. Macroaggregates containing attached fluorescently labeled bacteria (FLB) were obtained by adding labeled planktonic bacteria to rolling tanks filled with natural seawater, and they were used to measure protistan grazing rates on the bacteria attached to aggregates. Protistan grazing on free-suspended bacteria was measured according to the monodispersed FLB uptake technique. The four protists showed low grazing rates when feeding on free-suspended bacteria at the low densities usually found in seawater. Surprisingly, the analyzed protists also showed low grazing rates when feeding on aggregates despite the very high bacterial densities inhabiting them. The analysis of the influence of prey density on bacterial grazing rates suggests that feeding on bacteria attached to aggregates involves a relevant additional effort when compared to feeding on free-suspended bacteria. The paradox between these results and the very high abundance of protistan communities usually found in aggregates is discussed attending to the view of aggregates as biological reactors. Therefore, marine macroaggregates can be considered as microniches able to support the maintenance of a protistan community without food limitation, as well as to attract protistan colonizers by generating a water layer enriched in organic matter and bacteria.

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