Promastigotes of Leishmania braziliensis, L. donovani, L. major and L. mexicana recently derived from tissue amastigotes were cultured in Schneider's Drosophila medium supplemented with 20% (v/v) heat-inactivated foetal calf serum and 25 μg gentamicin sulfate/ml at pH 5·5. These cultures produced more metacyclic promastigotes in their stationary-phase populations than others cultured at pH 7·0. Metacyclic promastigotes possessed a short ([les ]8 μm) and narrow ([les ]1·5 μm) cell body with a flagellum twice or more the length of the cell body. Promastigotes from acidic cultures were more resistant to complement-mediated lysis and more infective in vivo than those grown at neutral pH. These results demonstrate that induction of metacyclogenesis by acidic pH is a response conserved across a variety of species of Leishmania.