The prevalence of antibody titers to Toxoplasma gondii in river otters (Lutra canadensis) from eastern North Carolina (USA) was investigated in a cross sectional study. Sera from 103 live trapped river otters were tested for antibodies to T. gondii using a commercially available latex agglutination kit. Forty-six (45%) of the sera were positive with titers ranging from 1:16 to >1:2,048. Adult otters (n = 78) had a seroprevalence of 47% and juvenile otters (n = 25) had a seroprevalence of 39%. Significant differences were not found between the sexes within either maturity class. The high prevalence of antibodies to T. gondii indicates that many animals in this population were exposed to the organism. This suggests handling of otters may be a zoonotic concern for fur trappers.