The fungus Neozygites floridana is a pathogen commonly associated with the cassava green mite (CGM), Mononychellus tanajoa, and the twospotted spider mite (TSM), Tetranychus urticae, in South America. Difficulty in culturing N. floridana in vitro has hampered its development as a bioacaricide. A Colombian isolate from TSM was multiplied in TSM and hyphal bodies from infected specimens were inoculated into 18 mycological and tissue culture media. Grace's insect tissue culture medium plus 0.33% lactalbumin hydrolysate and 0.33% yeastolate (GS) afforded vigorous multiplication of hyphal bodies (over 106/mL). The number of hyphal bodies increased with aeration. Final concentration of hyphal bodies was not affected by fetal bovine serum (FBS), but growth was somewhat enhanced. Hyphal bodies produced germ tubes when transferred from GS medium to Grace's medium plus 10% FBS and shaken at 200 rpm for 24 h. The germinated hyphal bodies, when placed on water agar, produced primary conidia from which capilliconidia were produced. These capilliconidia were infective to both TSM and CGM.