Physiological action of dissolved organic matter in rainbow trout in the presence and absence of copper: sodium uptake kinetics and unidirectional flux rates in hard and softwater.

We investigated the physiological effects of dissolved organic matter (DOM) on sodium (Na+) transport in juvenile Oncorhynchus mykiss (approximately 2.5 g) in the presence and absence of simultaneous acute exposure to copper (Cu2+; 0, 70, and 300 microg l(-1)). Trout were acclimated in either hardwater (approximately 1000 microM Ca2+) or softwater (approximately 100 microM Ca2+), and DOM was tested at approximately 8 mg C l(-1) using a natural (NOM) and a commercial (AHA) source. Ion transport was evaluated based on kinetics estimates (maximum Na+ uptake rates, Jmax; substrate affinity, Km) and unidirectional flux measurements (Jin, Jout, Jnet). Jmax was higher and unidirectional flux rates were greater in softwater-acclimated trout. Fish exposed to DOM alone in hardwater exhibited an increased Na+ transport capacity indicated by both the kinetics (67% higher Jmax for AHA) and Jin measurements (153% higher for AHA and 125% higher for NOM). In softwater, the effects of DOM alone on kinetic parameters and unidirectional flux rates were negligible. Cu2+ affected Na+ uptake by a mixed-type inhibition (both non-competitive and competitive). In hardwater, only Km was increased (i.e., affinity decreased), whereas in softwater, Km was increased and Jmax was decreased, with more marked effects at the higher Cu2+ level. In hardwater, the stimulatory effect of AHA on Jmax persisted even in the presence of 300 microg l(-1) Cu2+, whereas both AHA and NOM prevented the increase in Km caused by Cu2+; these effects were reflected in Jin measurements. In softwater, AHA helped to protect against the increased Km caused by high Cu2+, but there was no protection against the inhibition of Jmax. Unidirectional flux measurements indicated that in softwater, Cu2+ inhibited Jin at 70 microg l(-1), whereas at 300 microg l(-1) Cu2+, Jout was also stimulated. Fish were more affected by Cu2+ in softwater, as indicated by the inability to control diffusive losses of Na+ and a reduced ability to take up Na+, but in the presence of DOM, losses were better controlled at the end of 6 h exposure. We conclude that DOM has direct effects on the gills, as well as protecting fish against acute Cu2+ toxicity. This occurs because DOM complexes Cu2+, and because it acts on the transport and permeability properties of the gills. These effects differ depending on both water hardness and the nature of the DOM source.