The present study was conducted to evaluate the effect of low-dose gamma irradiation (0, 1, 3 and 5 kGy) on fatty acid composition of Rainbow trout (Oncorhynchus mykiss) fillet. Among all of the fatty acids, oleic acid (C18:1) (with mean 33.50±3.02 g/100 g fatty acids) and myristoleic acid (C14:1) (with mean 0.41±0.26 g/100 g fatty acids) were the most predominant and the lowest fatty acids in all irradiated and non-irradiated samples, respectively. Statistical analysis showed that there were no significant differences (P>0.05) in level of all fatty acids, saturated fatty acids (SFA), unsaturated fatty acids (USFA), mono unsaturated fatty acids (MUFA) and poly unsaturated fatty acids (PUFA) between Rainbow trout fillet control and irradiated in 1, 3 and 5 kGy. Therefore irradiation process and different doses of irradiation in this study (1, 3 and 5 kGy) had no significant effect (P>0.05) on fatty acid composition.