Nitrogen fixation is often cited as an important nitrogen source for termites. However, the rate of nitrogen fixation between experiments and termite species is highly variable due to termite diversity and sampling limitations. New evidence suggests that subterranean termites (Coptotermes formosanus Shiraki) (Blattodea, Rhinotermitidae) seek alternative dietary nitrogen sources for colony growth and development. This study compares N2 fixation rates and nitrogenase gene expression rates to a diverse assemblage of feeding groups. It was observed that nitrogenase expression was much higher in one-piece nesting termites from the family Kalotermitidae which lack access to soil. On the other hand, subterranean termites from the family Rhinotermitidae fix nitrogen at much lower rates, not differing from ambient nitrogenase activity in soil.