An isolate of Fusarium sporotrichioides Sherbakoff obtained from hay from Kindersley, Saskatchewan, and producing metabolites toxic to insect larvae, rats, and mice was cultured on sterile rye grain and extracted with ethyl acetate. The most toxic fraction derived from column chromatography on silica gel was purified by thin-layer chromatography of silica gel and elution of the main band. This compound was identified as T-2 toxin by comparison with the pure chemical by thin-layer chromatography and, after crystallization, by determination of melting point, infrared spectrum, and nuclear magnetic resonance spectrum. This work demonstrates that strains of F. sporotichioides capable of producing T-2 toxin may be present on hay in western Canada.