The aim of the present study was to evaluate the occurrence of some avian Haemosporidia, Coxiella burnetii and Francisella tularensis in waterfowl from Tuscany wetlands. One-hundred and thirty-three samples of spleen were collected from regularly hunted wild birds belonging to 13 different waterfowl species. DNA extracted from each sample was submitted to PCR assays and sequencing to detect the pathogens. Thirty-three samples (24.81%) were positive with PCR for at least one pathogen: 23 (17.29%) for Leucocytozoon spp., 6 (4.51%) for Plasmodium spp., 4 (3%) for C. burnetii , 2 (1.5%) for Haemoproteus spp. No specific F. tularensis amplifications (0%) were detected. To the best of our knowledge, this study firstly reports data about haemosporidian and C. burnetii infections in waterfowl from Italy.