When the research on the ecology of monkeypox virus entered the latest stage in 1984, three groups of animals were considered priority candidates for maintenance of virus circulations in nature because of their relatively higher population density: terrestrial rodents, squirrels, and gregarious bats. Following the isolation of the virus from a wild squirrel in 1985 another survey was carried out in Bumba zone of Zaïre in January-February 1986, which included collection of animal samples, later on tested by WHO collaborating laboratories. No antibodies were found in 233 rodents tested. A high prevalence of monkeypox-specific antibodies (24.7%) found in 320 squirrels of Funisciurus anerythrus species suggested that these animals sustain virus transmission in the areas surrounding human settlements. A consistently high level of antibody prevalence found among squirrels Heliosciurus rufobrachium suggested that this species is also steadily involved in the transmission. The squirrels, the only mammals frequently infected by monkeypox in the areas of human activities, which are frequently trapped by the population, are obviously a major source of infection for human beings. Out of 39 primates collected during the survey from the primary forest, three were found seropositive for monkeypox. At the moment, however, it is difficult to say whether the primates play an important role in sustaining virus transmission in nature, or (like human beings) that they are an occasional host of the virus. Possibilities are discussed of monkeypox virus transmission being sustained by squirrels outside the areas of major human activities.