A spectrofluorometric assay for the estimation of the tremorgenic mycotoxin verruculogen in crude mycelial extract has been devised and used to determine concentrations as low as 0.2 microgram ml-1. Verruculogen production by Penicillium estinogenum has been extended from surface culture to submerged culture in 60 1 stirred fermenters, in which the maximum cell-associated mycotoxin yield [5 mg (100 ml culture)-1] was obtained within 7 d. It was found necessary to supplement the medium (Czapek Dox broth plus 0.5% yeast extract) with calcium chloride (2%) to induce profuse sporulation (2 X 10(7) conidia ml-1).