We provide ecological information on Bothrops neuwiedi pauloensis (Amaral, 1925) regarding geographical distribution, habitat use, biometry, feeding habits, and reproduction, based on field studies and analysis of 175 preserved specimens. Bothrops n. pauloensis is endemic to the Brazilian Cerrado, where it occurs in open, seasonally dry savannas. Sexes are dimorphic in body length, relative tail length and relative body mass, but not in relative head length or diet. A wide array of prey, from centipedes to rodents, is consumed, and there is an ontogenetic dietary shift from ectotherms to endotherms. Prey-predator mass ratios ranged from 0.006–0.571. Although prey mass increased with snake mass, large snakes also ate many small prey. There was no difference in relative prey mass between sexes. Reproduction was highly seasonal, with a long vitellogenic period. Embryos were found only from October to December (rainy season). Litter size ranged from 4–20, and was dependent on female size. Although B. n. pauloensis occupies seasonally dry savannas, its ecological characteristics are similar to those described for Bothrops species inhabiting forested habitats.