Glycosylated molecules expressed on the cell surface of Leishmania promastigotes contribute to the outcome of contact between the parasite and its invertebrate and vertebrate hosts. The expression of several such molecules is growth phase dependent. Information on the expression of carbohydrates by Leishmania of the Viannia subgenus (braziliensis complex), a widespread cause of morbidity in the Americas, is fragmentary. We have examined the relationship between growth phase and the expression of glycosylated surface structures in WHO reference strains of 3 species of the Viannia subgenus, i.e., L. panamensis, L. guyanensis, and L. braziliensis. Agglutination with lectins and the monoclonal antibody specific for the repeat unit of L. donovani lipophosphoglycan, CA7AE, distinguished logarithmic and stationary-phase promastigotes of all 3 species. Flow cytometry revealed increased heterogeneity and disparity in the expression of the repeat unit epitope in stationary-as compared to logarithmic-phase promastigotes. Biochemical analyses showed the LPG repeat unit of all 3 species reference strains to be constituted by mannose and galactose with little or no substitution and, hence, to be similar to the LPG of L. donovani. Initial quantitative analyses of L. braziliensis LPG indicated a 10-fold lower quantity of LPG in this species than L. donovani and an increase in the size of LPG in the stationary phase. These findings provide bases for isolating and biologically characterizing phenotypically distinct populations of promastigotes and for identifying molecular determinants of the host parasite-relationship among Leishmania Viannia.