The South American water snake Helicops leopardinus is very abundant in the Pantanal wetlands of Central Brazil. We studied the biology of this species based on specimens collected by local workers in the Paraguay River. Females attained greater body lengths and had larger heads than males, but the latter had longer tails. Helicops leopardinus fed on a wide variety of fishes (70% of the total prey items) and frogs, with fishes of the order Gymnotiformes (33.34%) and frogs of the family Hylidae (23.34%) representing the most common prey items. There was relatively low diet overlap between the sexes and between juveniles and adults. Mean litter size was 6.6 and ranged from 4–11. The reproductive cycle was seasonal, with birth occurring late in the wet season (simultaneous with the flooding of the Paraguay River), facilitating dispersal of young snakes by floating vegetation. Floating vegetation may be a critical habitat for H. leopardinus, and management plans for their maintenance need to be developed.