Aflatoxin B-1 (AFB(1)) and deoxynivalenol (DON) are important food-borne mycotoxins that have been implicated in animal and human health. In this study, individual and combinative effects of AFB(1) and DON were tested in primary hepatocytes of Cyprinus carpio. The results indicated that the combinative effects of AFB(1) and DON (0.01 mu g/mL AFB(1) and 0.25 mu g/mL DON; 0.02 mu g/mL AFB(1) and 0.25 mu g/mL DON; 0.02 mu g/mL AFB(1) and 0.5 mu g/mL DON) were higher than that of individual mycotoxin (P < 0.05). The activity of AST, ALT and LDH in cell supernatant was higher than that of control group (P < 0.05) when the mycotoxins were exposed to primary hepatocytes for 4 h. The decreased cell number was observed in tested group by inverted light microscopy. The mitochondrial swelling, endoplasmic reticulum dilation and a lot of lipid droplets were observed in primary hepatocytes by transmission electron microscope. Therefore, this combination was classified as an additive response of the two mycotoxins.