Background: Mealtime best practices for obesity prevention in child care have been developed from experimental studies and expert opinion. Our objective was to describe adherence to best practices in child care centers and to evaluate the association between mealtime practices and children's dietary intake. Methods: We conducted an observational study of 349 preschoolers, ages 36 to 72 months, from 30 child care centers in Cincinnati, Ohio (November 2009 to January 2011). Trained observers recorded providers' behaviors related to six mealtime best practice recommendations and documented children's intake (n = 60 group lunches). General linear mixed models were used to evaluate the association between practice use and children's total energy (caloric consumption) and fruit and vegetable consumption. Results: Adherence to individual mealtime best practices was variable (0%-77%). Staff sitting with children at lunch was associated with lower energy intake and higher vegetable intake. Staff eating some of the same foods was associated with higher energy intake and higher vegetable intake. Staff encouraging children to try new/less-favorite foods more than once was associated with lower fruit intake. Staff having general conversations with children (not addressed in recommendations) was associated with lower vegetable intake. Family-style meal service, staff talking about healthy foods, and staff helping children assess hunger before seconds were not significantly associated with intake. Conclusions: Few mealtime best practices were associated with dietary intake. Given the number of meals children consume in child care and the prevalence of childhood obesity, efforts to identify mealtime practices that improve children's dietary intake are crucial for obesity prevention. (author abstract)