The present study is part of the Flora of Espírito Santo project and aimed to survey and describe the morphological diversity of species of Bixaceae from Espírito Santo state, Brazil, as well as to provide data on geographical distribution, biology, and habitat of these species. To do so, fieldwork was carried out in several areas of Espírito Santo and the most important herbaria collections in the state were examined. Bixaceae in Espírito Santo is represented by the genus Bixa, with two species. Bixa atlantica, recently described and repeatedly misidentified as Bixa arborea, is a forest tree up to 20 m tall (rarely appearing as treelet or shrub in savannah-like vegetation), with spine fruits and lacking nectaries in the pedicel apex. Bixa orellana, on the other hand, is recognized as a shrub or treelet up to 5-6 m tall, with conspicuous nectaries in the pedicel apex. The species origin is uncertain, though it occurs sub-spontaneously in Espírito Santo state, where it is widely cultivated due to its value as a spice. In this paper we provide taxonomic descriptions, an identification key, geographic distribution maps, photographic plates and comments on the ecology, conservation, and recognition of both species.