Deforestation and conversion of native habitats to exotic pasture and crops, plus inefficient agricultural and cattle management practices, are placing great pressures on natural resources in the Pantanal and Cerrado. To prevent further deforestation and protect biodiversity, areas already developed for farming and ranching need to be managed more efficiently and profitably, so that economic incentives for additional deforestation are minimized. To that end, the Wildlife Conservation Society (WCS) has been working with rural community partners to promote best-management practices that optimize profitability and efficient use of developed lands, while minimizing pressures on natural resources. To improve pasture conditions and reduce cattle impacts, we evaluated the use of rotational grazing as a management tool by monitoring native pasture and cattle within continuous and rotational grazing areas on a southern Pantanal ranch. Monthly comparisons of the grazing systems showed that forage dry mass in the rotational area was greater compared to that of continuously grazed areas. After 17 months, mean cattle weights were 15% heavier and pregnancy rates 22% higher for the herd using the rotational system. Based on forage allowance estimates, the potential stocking rates of the rotational system were 2 to 6 times higher than rates typical of continuously grazed areas in the Pantanal. Results support the use of rotational grazing in native pasture areas of the Pantanal.