Blood samples were analyzed from 30 domestic cats (Felis domesticus) from the Petén region of Guatemala to determine the seroprevalence of common pathogens that may pose a potential risk to native wild felids. Eight of the cats had been vaccinated previously; however, owners were unable to fully describe the type of vaccine and date of administration. In addition, blood samples were obtained from two captive margays (Leopardus wiedii). Samples were tested for antibodies to feline immunodeficiency virus, Dirofilaria immitis, feline panleukopenia virus, feline herpesvirus, feline coronavirus, canine distemper virus, and Toxoplasma gondii and for feline leukemia virus (FeLV) antigen. Fifty percent or more of the cats sampled were seropositive for feline herpesvirus (22 of 30), feline panleukopenia (15 of 30), and T. gondii (16 of 30). Five cats were positive for FeLV antigen. Both margays were seropositive for feline coronavirus and one was strongly seropositive to T. gondii. All animals were seronegative for D. immitis. This survey provides preliminary information about feline diseases endemic to the Petén region.