The composition and content of fatty acids (FAs) in total lipids, triacylglycerols (TAG) and polar lipids (PL) in dominant groups of benthic invertebrates: gammarids (Gammaridae, Amphipoda), chironomid larvae (Chironomidae, Diptera), caddisfly larvae (Trichoptera) and mayfly larvae (Ephemeroptera) were studied in the Yenisei river. For the first time data on the FA composition of species belonging to Trichoptera (Insecta) are presented. The groups of aquatic insect larvae and gammarids weakly differed in total content of essential polyunsaturated fatty acids (PUFAs). Hence, the strong invasion of gammarids which occurred in the last decades in the Yenisei river should not result in a decrease in potential yield of essential PUFA in the ecosystem and corresponding decrease in food resource quality for fish in respect to PUFA content. Significant differences in biomarker FAs in TAG were found which correlated to specific food sources. Different levels of long-chain PUFA in PL of the invertebrates are discussed in relation to the genetic ability of particular taxa to form these FAs.