Laboratory studies were carried out to investigate the cultural characteristics of salmonid alphaviruses (SAV) from Atlantic salmon (AS, Salmo salar) and rainbow trout (RT, Oncorhynchus mykiss), particularly in relation to cell line and temperature. In an initial study, SAV was isolated from 12 viraemic sera and passaged in Chinook salmon embryo (CHSE-214) cells at 15 °C. Geometric mean titres (GMT) after initial isolation were found to be significantly higher (P < 0.05) relative to those after two or four passages. Primary isolation of SAV was conducted from 12 viraemic sera (six AS and six RT) in seven different cell lines at 15 °C: CHSE-214, rainbow trout gonad (RTG-2), TO (derived from Atlantic salmon head kidney leucocytes), salmon head kidney (SHK-1), blue fin-2 (BF-2), fat head minnow (FHM) and Epithelioma papulosum cyprini (EPC). Overall, significant differences were found between cell lines in both the numbers of strains where growth was detected and in the GMT obtained. For both AS and RT strains, GMT values were significantly (P < 0.01) higher in both TO and BF-2 cells relative to the others, including CHSE-214 and RTG-2, the cell lines conventionally used for SAV. The effects of temperature of incubation (4, 10, 15 and 20 °C) on growth in TO, CHSE-214 and RTG-2 were investigated. In TO and RTG-2 growth was optimal at 15 °C, whereas in CHSE-214 results at 10 and 15 °C were more similar. Little or no growth was detected at 4 or 20 °C.