We determined mercury (Hg) concentrations in feathers of osprey (Pandion haliaetus), both nestlings (n = 95) and adults (n = 110), across peninsular Florida and the Florida Keys during February–August 2014. Feathers plucked from nestlings, aged 3–7 weeks, contained Hg concentrations that ranged from 0.338 to 45.79 mg/kg and averaged 6.92 ± 7.58 mg/kg (mean ± 1SD). Feathers shed from adults contained significantly higher concentrations ranging from 0.375 to 93.65 mg/kg, with an average of 17.8 ± 16.1 mg/kg. These levels were in the upper range of previously reported feather Hg concentrations of osprey and clearly show that Florida continues to have Hg hotspots that are elevated compared with many other regions. While these concentrations did not exceed levels previously reported in osprey from heavily Hg contaminated areas that showed no evidence of reproductive impairments, we cannot rule out potential individual-level effects to highly exposed nestlings after fledging. Mercury concentrations in nestlings were highest in coastal habitats of Collier and Monroe counties, where ongoing declines in osprey populations also have been documented.