From August to December 2018, we collected blood samples from 98 individuals of 11 mammal species to examine seroprevalence of leptospirosis at the Sherburne Wildlife Management Area in central Louisiana, USA. Overall, 21.4% of individuals tested positive for antibodies of at least one Leptospira interrogans serovar and six individuals were reactive for two or more serovars. The most prevalent serovar we detected was serovar Bratislava (19.4%), followed by serovar Grippotyphosa (6.1%), serovar Icterohaemorrhagiae and serovar Pomona (2.0%), and serovar Canicola and serovar Hardjo (1.0%). We detected the highest prevalence in fox squirrels (Sciurus niger), hispid cotton rats (Sigmodon hispidus), and feral swine (Sus scrofa), with serovar Bratislava being the most reactive for these three species. Positive samples returned titer results of 100–400 for all species and serovars, with the exception of one feral swine that returned a titer of 1,600 to serovar Bratislava, indicating an active infection. Although the potential effects of leptospirosis on our study species remains unclear, our data contribute information necessary to understand and manage potential risks of Leptospira exposure to wildlife, domestic animals, and humans.