A direct oxidation fuel cell of this invention has at least one unit cell including: a membrane-electrode assembly including an electrolyte membrane sandwiched between an anode and a cathode, each of the anode and the cathode including a catalyst layer and a diffusion layer; an anode-side separator with a fuel flow channel for supplying a fuel to the anode; and a cathode-side separator with an oxidant flow channel for supplying an oxidant to the cathode. The catalyst layer of at least one of the anode and the cathode includes high-porosity regions and low-porosity regions, and the high-porosity regions and the low-porosity regions are arranged in a mixed configuration.