The dietary utilization of cyanobacterial carbon by fish communities is poorly understood. We examined the transfer of cyanobacterial carbon to fish in a eutrophic lake using fatty acid biomarkers and measuring the stable carbon isotope ratios of fatty acid and bulk nitrogen. We collected five species of fish (Hypomesus nipponensis, Carassius sp., Cyprinus carpio, Tridentiger brevispinis, and Gymnogobius castaneus) as well as the seston from June to November 2016 from Lake Hachiro, Japan. Cyanobacterial blooms were observed from August to October. From June to August, cyanobacterial fatty acid biomarkers (18:2ω6 and 18:3ω3) accounted for only 1.4-4.3% of total fatty acids in these fish species, indicating a low contribution of cyanobacteria to fish diets during this period. However, the contribution of the cyanobacterial fatty acid biomarkers in these fish species increased sharply in September (10.5-17.1%), except in second-year H. nipponensis. In September, the stable carbon isotope ratios of 18:3ω3 in these fish species were almost equivalent to those in the seston, which was primarily composed of cyanobacteria. The trophic positions of the collected fish species ranged from 1.6 to 3.4, based on their stable nitrogen isotope values, indicating that some fish ingested cyanobacteria directly, while others acquired cyanobacteria indirectly, through the food chain. These findings indicate that cyanobacterial carbon is transferred up the food chain in eutrophic lake ecosystems with cyanobacterial blooms.