1. Aquatic and terrestrial ecosystems differ fundamentally in the abundance of longchained polyunsaturated fatty acids (PUFAs), such as eicosapentaenoic acid (EPA; 20:5n-3), which are produced by aquatic algae, but only in low quantities by terrestrial plants. Aquatic insects, such as Chironomidae (non-biting midges) feed on algae during their larval stage, making them rich in EPA and therefore high-quality prey for insectivores after emergence. However, the magnitude of EPA subsidies from aquatic insects may be different among water bodies in response to abiotic (e.g. nutrient load) as well as biotic factors (e.g. food web structure).2. To test the predation effects of crucian carp (Carassius carassius), nutrient concentrations, and Chironomidae community composition on the fatty acid export from aquatic ecosystems, we conducted a 25-day experiment across 20 1,500-L mesocosms covering a total phosphorus (TP) gradient of 20-1,000 µg/L.3. Twice a week, we collected adult emerging Chironomidae and found differences in fatty acid composition in the two most abundant chironomid species emerging from the mesocosms. Two PUFAs, α-linolenic acid and EPA, contributed to most of the variation in Chironomidae fatty acid content across the nutrient gradient.Whereas the proportions of α-linolenic acid were positively correlated to the mesocosm TP concentration, we found a negative correlation for the proportions of Chironomidae EPA and mesocosm TP concentration. However, despite lower biomass-specific EPA content at higher TP, higher biomass of emerging Chironomidae at intermediate TP concentrations resulted in higher total export of PUFAs from water to land. 4. Predation pressure from carp decreased the biomass of emerging Chironomidae on average 8-fold. Chironomidae biomass showed a hump-shaped relationship along the TP-gradient and was strongly influenced by periphyton biomass. 5. Export rates of EPA and fatty acids in general responded in a quadratic manner along the nutrient gradient, reaching a maximum value at a TP of 400 µg/L and decreasing thereafter.