Rock materials are often affected by water in underground engineering. In this study, the mechanical and failure characteristics of tuff under different moisture content were studied using laboratory tests. The moisture content variation of tuff was studied in water absorption tests, and the mechanical and failure characteristics of tuff under different moisture contents were studied through uniaxial compression tests with a Micro-II acoustic emission (AE) control acquisition system. The results showed that the moisture content of tuff increases rapidly at the initial stage of water absorption tests and stabilizes after 180 h of immersion. According to the results of uniaxial compression tests, both uniaxial compressive strength and elasticity modulus decreased with the increase in moisture content. The AE parameters analyses showed that, when the moisture content increased, the accumulated AE counts and energy gradually decreased, and the “quiet period” at the initial stage of uniaxial compression tests lasted longer, and the RA (rise time⁄amplitude)–AF (AE counts⁄duration) distribution and the failure characteristics verified that the failure patterns evolved from shear failure to tensile failure. Scanning electron microscopy was used to observe the morphology of the fracture surface and analyze the influence of moisture content on the fracture characteristics of the tuff at the mesoscopic level. The results of this research can be used as a basis for studying the influence of water on tuff.