Intertidal harpacticoid copepods are commonly used in eco-toxicity tests worldwide. They predominately live in mid-high shore rock pools and often experience a wide range of temperature and salinity fluctuation. Most eco-toxicity tests are conducted at fixed temperature and salinity and thus the influence of these environmental factors on chemical toxicity is largely unknown. This study investigated the combined effect of temperature and salinity on the acute toxicity of the copepod Tigriopus japonicus against two common biocides, copper (Cu) and tributyltin (TBT) using a 2 x 3 x 4 factorial design (i.e. two temperatures: 25 and 35 degrees C; three salinities: 15.0 per thousand, 34.5 per thousand and 45.0 per thousand; three levels of the biocide plus a control). Copper sulphate and tributyltin chloride were used as the test chemicals while distilled water and acetone were utilised as solvents for Cu and TBT respectively. 96 h-LC50s of Cu and TBT were 1024 and 0.149 microg l(-1) respectively (at 25 degrees C; 34.5 per thousand) and, based on these results, nominal biocide concentrations of LC0 (i.e. control), LC30, LC50 and LC70 were employed. Analysis of Covariance using 'concentration' as the covariate and both 'temperature' and 'salinity' as fixed factors, showed a significant interaction between temperature and salinity effects for Cu, mortality increasing with temperature but decreasing with elevated salinity. A similar result was revealed for TBT. Both temperature and salinity are, therefore, important factors affecting the results of acute eco-toxicity tests using these marine copepods. We recommend that such eco-toxicity tests should be conducted at a range of environmentally realistic temperature/salinity regimes, as this will enhance the sensitivity of the test and improve the safety margin in line with the precautionary principle.