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The paleoenvironmental conditions surrounding the origins of pastoralism and the movement of herders from eastern to southern Africa sometime between similar to 4000 and 2000 ybp have been much debated. We lack, however, detailed paleoenvironmental data from sites sampling the hunter-to-herder transition in southeastern Africa, the likely corridor from eastern to southern Africa for early pastoralists. Here we report on new paleoenvironmental data from a site in the under-sampled area of eastern Zambia, Makwe Rockshelter, which has two aggregates of archaeological horizons representing the mid-Holocene (similar to 5700-5000 ybp) and the late Holocene (similar to 1600-800 ybp). The mid-Holocene sediments at Makwe document a foraging society, whereas the late Holocene sediments include both wild game and domestic livestock. Using stable carbon isotopes (delta C-13) of herbivore enamel (n = 107), we show that the shift from mid-Holocene to late Holocene paleoenvironments was characterized by an increase in C4 vegetation. These data are complemented by paleoenvironmental records from Lake Malawi that show that C4 vegetation peaked after similar to 2000 ybp and was coincident with the onset of cooler, more arid climates. This combined paleoenvironmental record has implications for the spread of pastoralism across southeastern Africa between similar to 3000 and 2000 ybp and potential ’animal disease barriers’ these early herders may have faced. (C) 2016 Elsevier Ltd. All rights reserved.