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Collagen films with proton conduction are a candidate of next generation of fuel-cell electrolyte. To clarify a relation between proton conductivity and formation of water networks in the collagen film originating from a tilapia's scale, we systematically measured the ac conductivity, infrared absorption spectrum, and weight change as a function of relative humidity (RH) at room temperature. The integrated absorbance concerning an O-H stretching mode of water molecules increases above 60% RH in accordance with the weight change. The dc