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Several factors which influence Rickettsia tsutsugamushi infectivity of, and replication in, cultured cells have been investigated in order to establish standard laboratory conditions for reliable quantitation and maximization of rickettsial yield and to begin to explore the nature of the relationship between the rickettsia and its host cell. The extent of scrub typhus rickettsial association with host cells was dependent on the rickettsial and target cell concentrations and on the medium in which the infection took place. Brain heart infusion broth, a standard rickettsial diluent for infectivity, was markedly inhibitory. Both rickettsial and host cell replication were dependent on a component(s) supplied by serum to the tissue culture medium; rickettsial multiplication was less affected by small temperature variation (optimum 34 to 37°C) than was host cell growth (optimum 37°C). The antibiotic daunomycin was very useful in selectively inhibiting host cell replication and permitting attainment of somewhat greater rickettsial yields than in uninhibited cells. These findings have underscored the close relationship between parasite and infected cell and have led to more predictable, higher yields of rickettsiae in vitro.