To determine the relative importance of two main factors, diet or culture environment, that affect the proximate composition and main nutritive ingredients (vitamin A, vitamin E, cholesterol, fatty acid composition) in cultured eels, we analyzed the composition of eels fed diets of formula feed (FF) produced by four different companies and of eels cultured at five different eel farms that provided only one of the four different FFs. The four commercial eel FFs did not markedly differ in proximate composition or major nutritive compounds, and consequently, these variables did not significantly differ in cultured eels fed the different FFs. The FF imported from Japan was marginally superior to the two domestic commercial FFs and the FF imported from Taiwan in terms of the proximate composition and main nutritive ingredients of both the FF itself and the eels cultured on it. However, proximate composition and main nutritive ingredients significantly differed among eels cultured at the five farms that used a different FF and among eels fed the four different FFs at the same farm. In conclusion, the difference in quality between domestic and Japanese FFs in terms of eel culture was small, whereas physical or chemical environmental differences among farms during eel culture may more strongly affect the proximate composition and levels of the main nutritive ingredients in cultured eels.