The burden of rotavirus disease in the Latin American region has been poorly understood despite the promise of effective vaccines. We describe here the implementation and results of a rotavirus surveillance network in the Latin American and Caribbean region. From 2005 through 2007, stool specimens and epidemiologic information were gathered from children <5 years of age who were hospitalized for acute diarrhea (3 looser-than-normal stools within <24 h) lasting <14 days with use of a standardized generic protocol. Stool samples were tested for rotavirus, and a proportion of detected strains were typed. The proportion of samples positive for rotavirus was applied to World Health Organization diarrhea-related mortality estimates to calculate rotavirus-associated mortality. In 2007, the network comprised 54 sites in 11 countries. During 2006-2007, specimens were collected from 19,817 children; 8141 of these specimens were positive for rotavirus. The median percentage of positive specimens in the country was 31.5% (range, 24%-47%). The risk of death from rotavirus diarrhea by age 5 years was 1 of 2874. Strong rotavirus winter seasonality was apparent, even in tropical Central America. Globally common strains (P[8] G1, P[8] G9, and P[4] G2) accounted for >75% of strains, although unusual strains, including G12, were detected at low levels. As rotavirus vaccines continue to be introduced in Latin America, maintenance of surveillance will provide robust pre-introduction data and a platform for estimating vaccine effectiveness and other measures of impact.