Co-cultivation of Legionella pneumophila serogroup I and Acanthamoeba palestinensis in Neff's medium at 35 degrees C resulted in the intracellular multiplication of the bacteria as demonstrated by electron microscopy and immunofluorescence. In the closed experimental system used, the number of legionellae rose from 10(7) colony forming units (c.f.u.)/ml initially to a maximum of 10(10) c.f.u./ml on day 5. Legionellae were seen in expelled phagosomes, in some amoebae filling the cytoplasm and in others in which the process of encystment appeared to have commenced. At 20 degrees C the acanthamoebae phagocytosed and digested the legionellae. The bacteria disappeared from the co-cultivation flask by day 2 but reappeared in low numbers (10(2) c.f.u./ml) by day 6 suggesting that even at this temperature some intra-amoebal multiplication occurred.