Wild and feral medium-sized mammals were live trapped at two natural protected areas within the Mexico City limits to determine antibody prevalence for the most common infectious diseases (rabies, toxoplasmosis, and canine parvovirus) in dogs and cats. Mammals were trapped during the dry (March–April) and rainy seasons (July– August) of 1996 and 1997. A total of 68 individuals were captured, representing 8 species: opossums (Didelphis virginiana), ringtails (Bassariscus astutus), spotted skunks (Spilogale gracilis), weasels (Mustela frenata), rock squirrels (Spermophilus variegatus), Mexican gray squirrels (Sciurus aureogaster), feral cats (Felis catus), and feral dogs (Canis familiaris). There was marked seroprevalence for parvovirus (86.6%) and lower seroprevalences for both toxoplasma (23.9%) and rabies (17.9%). There were no significant prevalence differences among mammals in both protected areas, which were of contrasting size and isolation (i.e., small and isolated versus large and nonisolated). We suggest that high seroprevalence of these three infectious agents in wild mammals is a result of the high densities of feral dogs and cats in the two areas sampled. Feral dogs are able to maintain the infectious agents in these localities regardless of the protected area size and isolation. However, the native mammals of the small and isolated reserve are more vulnerable to infectious diseases because of small population size and genetic bottlenecks. Our results indicate that natural areas in and around Mexico City are a refugium for latent infectious agents, several of which are zoonotic. These findings suggest that conservation measures, such as eradication of feral mammals and vaccination programs, in the protected areas and surrounding areas could be beneficial.