The invention provides a single-chamber microbial fuel cell, which takes a gas diffusion electrode as the negative electrode. The invention comprises a container, a positive electrode and a negative electrode; wherein, the positive electrode adopts a foamed metal electrode, the negative electrode adopts a gas diffusion electrode containing metallic catalyst, such as Ag, the container adopts a cylindric glass container, a water inlet opening is arranged at the lower end of the container, a water outlet opening is arranged at the upper end, a sealing cover is arranged at the upper part of the container, a sampling opening is arranged at the middle part of the sealing cover, the positive electrode is positioned at the inner side of the container, the negative electrode is positioned at the outer side of the container, the positive electrode and the negative electrode are arranged along the circumference of the container, copper conductors are connected between the both electrodes, and the both electrodes are connected with a load to form a closed loop. The gas diffusion electrode adopted by the invention exposes in the air, the oxygen gas in the air can be directly utilized as electron acceptors, an extra aerating device is not required; therefore the running cost is reduced. Organism, such as dextrose and discharge water can be used as the fuel of the cell, chemical energy can be effectively converted into electric energy; simultaneously, better discharge water processing effect can be achieved.