Feline hemoplasmas, consisting of Mycoplasma haemofelis (M. haemofelis), Candidatus Mycoplasma haemominutum (Ca. M. haemominutum), and Candidatus Mycoplasma turicensis (Ca. M. turicensis), cause feline infectious anemia and zoonoses. Using multiplex PCR and phylogenetic analysis based on 16S rRNA, 22 blood samples from fishing cats (Prionailurus viverrinus) living in Khao Sam Roi Yot National Park, Thailand were determined positive for M. haemofelis (13.6%)and Ca. M. haemominutum (22.7%). M. haemofelis and Ca. M. haemominutum infection can result severe anemia and asymptomatic, respectively. However, not all positive cases exhibit anemia symptoms. Future study of hemoplasma infection in wild felids is necessary for conservation and the preservation of public health in Thailand.