Mycoplasma iguanae proposed species nova was isolated from vertebral abscesses of two feral iguanas (Iguana iguana) from Florida. Three strains were evaluated for sensitivity to a variety of antibiotics. The minimum inhibitory concentrations for M. iguanae, assessed by broth dilution methods, of clindamycin, doxycycline, enrofloxacin, oxytetracycline, and tylosin (all <1 μg/ml) were lower than those of chloramphenicol (32 μg/ml) and erythromycin (64 μg/ml). The profile was identical to that of Mycoplasma alligatoris, previously isolated from American alligators (Alligator mississippiensis). M. iguanae strain 2327^T was subcultured without antibiotics to assess mycoplasmacidal activity. Clindamycin, doxycycline, oxytetracycline, and tylosin were bacteriostatic from 0.1 to 0.5 μg/ml, whereas enrofloxacin was bactericidal at 20 ng/ml. An enrofloxacin dosage of 5–10 mg/kg achieves peak plasma concentrations >1 μg/ml, with an elimination half-life of 6–20 hr, in alligators. Although concentrations achieved in the vertebrae by i.m. or i.v. injection are probably lower than those in plasma, these data suggest that enrofloxacin may be useful to treat M. iguanae mycoplasmosis in iguanas.