Acidic glycosphingolipid components were extracted from the mycelium form of the thermally dimorphic mycopathogen Sporothrix schenckii. Two fractions from the mycelium form (Ss-M1 and Ss-M2), having the highest Rf values on HPTLC analysis, were isolated and their structures elucidated by 1- and 2-D C-13- and H-1-nuclear magnetic resonance spectroscopy, and electrospray ionization mass spectrometry with lithium adduction of molecular ions, The structures of Ss-M1 and Ss-M2 were determined to be Man alpha1-->Ins1-P-1Cer and Man alpha1-->3Man alpha1-->Ins1-P-1Cer, respectively (where Ins = myoinositol, P = phosphodiester). The Man alpha1-->6Ins motif is found normally in diacylglycerol-based glycophosphatidylinositols of Mycobacteria, but this is the first unambiguous identification of the same Linkage making up the core structure of fungal glycosylinositol phosphoryl-ceramides (GIPCs). These results are discussed in relation to the structures of GIPCs of other mycopathogens, including Histoplasma capsulatum and Paracoccidioides brasiliensis.