Hirtusneanoside, a new O-deoxyglycoside of a dimeric tetrahydroxanthone, was isolated from the lichen Usnea hirta. Its structure, including the absolute configuration, was determined by means of extensive spectroscopic data (UV, IR, MS, CD, 1D and 2D NMR) and chemical degradation. Hirtusneanoside has a unique structure comprising L-rhamnopyranoside of a tetrahydroxanthone dimer and showed growth inhibitory activities against Gram-positive bacteria.