Aflatoxin contamination of foods and livestock feeds is an ongoing problem. In this research, NovaSil PLUS (NSP), a calcium montmorillonite clay that is commonly used as an anticaking agent in feeds, was evaluated for its ability to bind aflatoxin B1 (AfB1) in vitro and to prevent the onset of aflatoxicosis and vitamin A depletion in broiler chicks in vivo. Isothermal analyses were conducted with NSP and AfB1 to quantitate and characterize critical sorption parameters at equilibrium, i.e., ligand saturation capacities, affinity constants, and thermodynamics of the sorption process. In vitro results indicated that AfB1 was tightly sorbed onto the surface of NSP, which provided a high capacity and high affinity for the ligand. Thermodynamics favored sorption of AfB1 to NSP. The process was exothermic and spontaneous with a mean heat of sorption equal to approximately -50 kJ/mol, suggesting chemisorption (or tight binding). In addition to the in vitro studies, the effectiveness of NSP as an aflatoxin enterosorbent to attenuate the onset of aflatoxicosis in broiler chicks was determined at 3 different inclusion levels in the diet (0.5, 0.25, and 0.125%). NSP alone was not toxic to chicks at a level as high as 0.5% in the total diets (based on body and organ weights, feed intake, and hepatic vitamin A levels). NSP in the diet significantly protected chicks from the effects of highlevel exposure to aflatoxins (i.e., 5 mg/kg) and preserved hepatic vitamin A levels, even at lower dietary intake of clay.