Japanese climbing fern (Lygodium japonicum) is an invasive vine that has been identified as an economic and ecological threat in forest ecosystems of the Southeast. In two separate studies, we examined the use of directed sprays of glyphosate, imazapyr, and metsulfuron-methyl herbicides, alone and in combination, for control of Japanese climbing fern and for impacts to associated vegetation in mixed bottomland hardwood–cypress forests at two first terrace sites of the Apalachicola River in north Florida. Two yr after treatment, various rates of glyphosate alone generally provided greater percent control (84 to 95%) than imazapyr alone (−25 to 86%) or metsulfuron-methyl alone (−5 to 53%). Combinations of 2% glyphosate and imazapyr and/or metsulfuron-methyl resulted in 81 to 97% control at 2 yr after treatment, with no significant differences among the combination treatments or 2% glyphosate alone. Change in percent cover of associated vegetation groups was not influenced by treatment except for one location where graminoid cover increased 1 yr after metsulfuron-methyl treatment but decreased after imazapyr or glyphosate application. Species richness of dominant understory vegetation remained nearly constant at both locations regardless of treatment. These data indicate that glyphosate alone or in combination with imazapyr or metsulfuron-methyl will provide effective, although not complete, control of Japanese climbing fern, and that directed herbicide sprays minimally impact nontarget vegetation.