We investigate tracer diffusion at the domain boundaries in an adsorption layer, an effect that corresponds to grain boundary diffusion in 3D polycrystalline solids. Experiments were performed on adsorbed O atoms on a Ru(0001) surface in a layer of CO molecules. The CO molecules form a (3x3)R30∘ structure which displays translational domains. High-speed scanning tunneling microscopy (STM) was used to image the motion of the O atoms. The data show that single O atoms preferentially move along the domain walls which in the STM movies appear as disordered, fluctuating stripes between the ordered domains. The diffusion coefficient of the O atoms is one order of magnitude higher than the diffusion coefficient in the ordered domains. By comparison with previous experiments on completely disordered CO layers, it is concluded that the diffusion is similarly promoted by the enhanced fluctuations in the disordered domain walls.