Although urea production and metabolism in lungfish have been thoroughly studied, we have little knowledge of how internal osmotic and electrolyte balance are controlled during estivation or in water. We tested the hypothesis that, compared with the body surface of teleosts, the slender African lungfish (Protopterus dolloi) body surface was relatively impermeable to water, Na ϩ , and Cl Ϫ due to its greatly reduced gills. Accordingly, we measured the tritiated water ( 3 H-H 2 O) flux in P. dolloi in water and during air exposure. In water, 3 H-H 2 O efflux was comparable with the lowest measurements reported in freshwater teleosts, with a rate constant (K) of 17.6% body water h Ϫ1 . Unidirectional ion fluxes, measured using 22 Na ϩ and 36 Cl Ϫ , indicated that Na ϩ and Cl Ϫ influx was more than 90% lower than values reported in most freshwater teleosts. During air exposure, a cocoon formed within 1 wk that completely covered the dorsolateral body surface. However, there were no disturbances to blood osmotic or ion (Na ϩ , Cl Ϫ ) balance, despite seven-to eightfold increases in plasma urea after 20 wk. Up to 13-fold increases in muscle urea (on a dry-weight basis) were the likely explanation for the 56% increase in muscle water * Corresponding author. Address for correspondence: Department of Biology, Wilfrid Laurier University, 75 University Avenue West, Waterloo, Ontario N2L 3C5, Canada; e-mail: mwilkie@wlu.ca. content observed after 20 wk of air exposure. The possibility that muscle acted as a "water reservoir" during air exposure was supported by the 20% decline in body mass observed during subsequent reimmersion in water. This decline in body mass was equivalent to 28 mL water in a 100-g animal and was very close to the calculated net water gain (approximately 32 mL) observed during the 20-wk period of air exposure. Tritiated water and unidirectional ion fluxes on air-exposed lungfish revealed that the majority of water and ion exchange was via the ventral body surface at rates that were initially similar to aquatic rates. The 3 H-H 2 O flux declined over time but increased upon reimmersion. We conclude that the slender lungfish body surface, including the gills, has relatively low permeability to water and ions but that the ventral surface is an important site of osmoregulation and ionoregulation. We further propose that an amphibian-like combination of ventral skin water and ion permeability, plus internal urea accumulation during air exposure, allows P. dolloi to extract water from its surroundings and to store water in the muscle when the water supply becomes limited. Physiological and Biochemical Zoology

A time course analysis using (110m)Ag, (24)Na(+), and (36)Cl(-) examined gill silver accumulation and the mechanism by which waterborne silver (4.0 x 10(-8) M; 4.3 microg/l) inhibits Na(+) and Cl(-) uptake in gills of freshwater rainbow trout. Analyses of gill and body fluxes allowed calculation of apical uptake and basolateral export rates for silver, Na(+), and Cl(-). To avoid changes in silver bioavailability, flow-through conditions were used to limit the buildup of organic matter in the exposure water. For both Na(+) and Cl(-) uptake, apical entry, rather than basolateral export, was the rate-limiting step; Na(+) and Cl(-) uptake declined simultaneously and equally initially, with both uptakes reduced by approximately 500 nmol.g(-1).h(-1) over the 1st h of silver exposure. There was a further progressive decline in Na(+) uptake until 24 h. Carbonic anhydrase activity was inhibited by 1 h, whereas Na(+)-K(+)-ATPase activity was not significantly inhibited until 24 h of exposure. These results indicate that carbonic anhydrase inhibition can explain the early decline in Na(+) and Cl(-) uptake, whereas the later decline is probably related to Na(+)-K(+)-ATPase blockade. Contrary to previous reports, gill silver accumulation increased steadily to a plateau. Despite the rapid inhibition of apical Na(+) and Cl(-) uptake, apical silver uptake (and basolateral export) increased until 10 h, before decreasing thereafter. Thus silver did not inhibit its own apical uptake in the short term. These results suggest that reduced silver bioavailability is the mechanism behind the pattern of peak and decline in gill silver accumulation previously reported for static exposures to silver.

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Rainbow trout were exposed to a range of silver concentrations (as AgNO3) in flowing synthetic soft water (0.05 mM Na+, 0.05 mM Cl-, 0.05 mM Ca2+, 0.02 mM Mg2+, 0.02 mM K+, pH 7.0, approximately 0.7 mg C/L dissolved organic carbon, 10 mg CaCO3/L, 10 +/- 2 degrees C) to investigate a possible relationship between short-term gill silver accumulation (3 h or 24 h) and acute silver toxicity (96-h mortality). We also investigated potential relationships between gill silver accumulation and inhibition of Na+ uptake plus inhibition of gill Na+K(+)-adenosine triphosphatase (ATPase) activity. The 96-h median lethal concentration (LC50) values were 13.3 microg total Ag L(-1) and 3.3 microg dissolved Ag L(-1). A relationship was demonstrated between 3-h and 24-h gill silver accumulation and 96-h mortality. A relationship also was demonstrated between gill silver accumulation and inhibition of Na+ uptake at 24 h of exposure. No relationship between gill silver accumulation and inhibition of gill Na+K(+)-ATPase activity was found. The 96-h median lethal gill accumulation (LA50) values of 129 (at 3 h) and 191 ng g(-1) (at 24 h) and a conditional equilibrium binding constant of 8.0 for Ag+ binding to the gills were calculated. These observations support use of the silver biotic ligand model (BLM) as a regulatory tool to predict acute silver toxicity.

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