Chemical and biological responses to simultaneous additions of acid, aluminium and lime were investigated in contiguous 250m-reaches of a chronically acidic stream in Wales. Treatments were applied for 24 h, and from the upstream end were as follows: zone A-untreated, pH 5.0, 0.37 mg litre(-1) filterable Al; zone B-acidified to pH 4.5, 0.40 mg Al litre(-1) (47% of Al attributed to release from the stream bed due to acid additions); zone C-acidified to pH 4.5 and Al dosed to 0.67 mg litre(-1); zone D-dosed with limestone slurry, resulting in pH 7.2, 0.13 mg Al litre(-1). In all reaches, the chemistry of the interstitial water at depths of 0.15 and 0.3 m never fell below pH 5.5, with corresponding decreases in Al and increases in base cation concentrations. Brown trout, Salmo trutta, and crayfish, Austropotamobius pallipes, held in the stream showed decreases in plasma [Na(+)] and haemolymph [Na(+)], respectively, in all acidic zones (A, B, C): these responses were mitigated by liming (zone D). Thus both chronic and simulated episodic levels of pH and dissolved Al were sub-lethally toxic to test species of aquatic fauna. This experiment also demonstrates a stream bed source/sink of Al, and the availability of a possible refuge from acidic surface waters within the substratum.