Cadmium forms neutral, lipophilic Cd(L)2(0) complexes with diethyldithiocarbamate (DDC) and with ethylxanthate (XANT). Uptake of these complexes bythree unicellularfreshwater green algae (Chlamydomonas reinhardtii, Chlorella fusca, and Pseudokirchneriella subcapitata) was determined at two pH values (7.0 and 5.5) and compared to uptake of the free, uncomplexed Cd2+ ion. Uptake of the lipophilic complexes over time, characterized by high initial uptake rates but tending toward a plateau after about 30 min, could be modeled successfully as the result of the following processes: first-order uptake from solution, partitioning of the accumulated Cd into two internal pools (labile and nonlabile), and first-order loss of Cd from the labile pool. At pH 7.0 initial uptake rates for both Cd(L)2(0) complexes were much higher than for Cd2+ alone (e.g., up to approximately 90 times higher for comparable dissolved Cd concentrations of approximately 0.4 nM). However, the initial uptake rates for the lipophilic complexes dropped dramatically when the pH was lowered from 7.0 to 5.5 (2- to 60-fold decrease, depending on the algal species and the nature of the neutral complex). Loss rates for the accumulated complexes also decreased atthe lower pH. The lipophilicity of the neutral complexes, as estimated from their octanol-water distribution coefficient (Dow), was not affected by the decrease in pH from 7.0 to 5.5. We thus conclude that the acidification of the external medium, i.e., the interaction of protons with the algal membrane, strongly affects algal membrane permeability.

Uptake of lipophilic metal complexes by freshwater algae has recently been shown to be pH dependent. Here we look at different physiological aspects that could influence the diffusion of the lipophilic Cd complex, Cd(diethyldithiocarbamate)2 (0) (Cd(DDC)2 (0) ), into algal cells at different exposure pH values. Changes in cell membrane permeability were assessed as a function of pH for three species of green algae [Chlamydomonas reinhardtii P. A. Dang., Pseudokirchneriella subcapitata (Korshikov) Hindák, and Chlorella fusca var. vacuolata Shihira et R. W. Kraus] using two neutral, nonionic probes, fluorescein diacetate (FDA) and D-sorbitol. In parallel experiments, we exposed algae to inorganic Cd or to Cd(DDC)2 (0) and monitored Cd intracellular metal distribution, together with phytochelatin synthesis. For the three algal species acclimated at pH 5.5 (w/wo DDC 1 μM) and exposed at this pH, their permeability to FDA and D-sorbitol was consistently lower than for algae growing at pH 7.0 and exposed at this pH (P < 0.001). The ratio of the FDA hydrolysis rate measured at pH 7.0 with respect to the rate measured at pH 5.5 (both in the presence of DDC) correlated with the ratio of the Cd(DDC)2 (0) initial internalization rate constant obtained at pH 7.0 versus that obtained at pH 5.5 (three algae species, n = 9, r = 0.85, P = 0.004). Our results strongly suggest that acidification affects metal availability to algae not only by proton inhibition of facilitated metal uptake but also by affecting membrane permeability.

We have examined the influence of pH and a natural humic acid on the short-term uptake (<40 min) of a neutral, lipophilic metal complex by a unicellular freshwater alga, Pseudokirchneriella subcapitata. Cadmium diethyldithiocarbamate ([Cd(DDC)2]0) was used as a model lipophilic metal complex and Suwannee River Humic Acid (SRHA) was chosen as a representative aquatic humic acid (6.5 mg C L−1). Under the experimental conditions virtually all the Cd was expected to be present as the lipophilic complex ([Cd]T = 0.38 nM; [DDC] 1 μM; [Cd2+] <10−15 M; pH 7.0, 6.0, or 5.5). Uptake of [Cd(DDC)2]0 proved to be sensitive to pH changes. It was lower at pH 6.0 and 5.5 than at pH 7.0. To our knowledge, this is the first demonstration of reduced uptake of a lipophilic metal complex at low pH. The presence of SRHA also affected uptake, either by binding the lipophilic complex in solution and reducing its bioavailability (pH 7.0) or by increasing the permeability of the algal membrane (pH 5.5).