Background: Appetite traits and feeding practices have been linked to children's weight status. For example, eating in response to food cues (high food responsiveness (FR)), and poor regulation of intake (low satiety responsiveness (SR)), increase risk for obesity. Appetitive traits of infants, and feeding practices, are typically measured by parent-report. The purpose of this study was to use a modified eating in the absence of hunger (EAH) paradigm, measuring infants' intake 30 min after a typical meal, to test whether infant acceptance of a second meal is associated with parent-reported appetitive traits or feeding practices. Methods: Healthy infants aged 3–5 months (N = 54) were fed a typical meal and then offered a second meal by bottle 30 min later. Appetitive traits and feeding practices were assessed with surveys. Analyses of covariance were used to assess whether appetitive traits differed by acceptance of the second meal after adjusting for covariates. Results: Fifty-nine percent of infants accepted the second meal and these infants had greater parent-reported FR (M = 3.06 ± 0.58 vs M = 2.43 ± 0.80, p < 0.01) and lower SR (M = 1.87 ± 0.62 vs M = 2.67 ± 0.87, p = 0.01), compared to infants who rejected it. Group differences remained after adjusting for infant age, feeding mode, weight-for-length, and maternal body mass index. No other appetitive traits or feeding practices differed by group. Conclusions: Results expand research in older children by showing that infant response to a modified EAH protocol is associated with parent-reported FR and SR. Future research with this protocol in infants should investigate the consistency of this behavior across time and examine whether response to this protocol predicts subsequent growth.