The role of the macrophage in destruction of virulent treponemes is disputed. A major obstacle has been the inability to demonstrate quantitative phagocytosis of treponemes by macrophages. Treponema pallidum pertenue was attached to polycarbonate filters for assessment of treponemal phagocytosis by macrophages. The disappearance of treponemes due to phagocytosis was measured by enumeration with immunofluorescence. Resident and lipopolysaccharide-activated macrophages were found to phagocytize treponemes equally well. The phagocytosis of killed treponemes by macrophages was enhanced by opsonization with immune serum. Macrophages successfully phagocytized Staphylococcus aureus organisms when they were incubated on filters under identical conditions. Treatment of macrophages with cytochalasin B, a known inhibitor of phagocytosis, prevented the disappearance of treponemes and phagocytosis of S. aureus. In addition, fluorescent treponemal debris was observed only inside macrophages cultured with treponemes. These results demonstrate that macrophages can phagocytize pathogenic treponemes on polycarbonate filters.