Groundwater samples in Horonobe district, Hokkaido, were analyzed to evaluate the possibility that formic and acetic acids are active substrates for methanogens in Quaternary and Neogene (Koetoi formation) formations. ΔG_r corresponding to CH₄-producing reactions indicates that both acids could be active substrates in almost all sampling locations. However, acetic acid was recognized to be an active substrate only in the Koetoi formation on the basis of the principle of competitive exclusion (CE) of microorganisms. The limited possibility by the CE principle suggests that dynamic equilibrium between substrate production rates and consumption rates is established only in the Koetoi formation for acetic acid.