Simple SummarySodium butyrate is an important nutritional additive for livestock animals and its potential for improving growth performance is of great interest. The present study reported the optimal sodium butyrate supplementation level in milk for preweaning dairy calves and the effects of sodium butyrate supplementation in milk on the growth performance and intestinal microbiota of calves around the newborn stage and provided evidence of the effects of sodium butyrate supplementation in milk for dairy calf feeding.AbstractThe aim of the present study was to investigate the effects of sodium butyrate (SB) supplementation on the growth and intestinal microbiota of preweaning dairy calves. Eighty newborn Holstein calves (56 female and 24 male) were randomly allocated to four treatment groups with 20 calves each (14 female and 6 male). The suckling milk for the four treatments was supplemented with 0, 4.4, 8.8, or 17.6 g/d SB. During the 6-week experiment, dry matter intake was recorded daily, body weight was measured weekly, and rectal fecal samples were collected in the 2nd week. The V3–V4 hypervariable regions of the microbial 16S rRNA were amplified and then sequenced. SB supplementation elevated average daily gains (ADGs) in the first and second weeks. The optimal SB supplementation level for the whole preweaning period was 8.78 g/d, as revealed by analyzing the whole preweaning period ADG using second-order polynomial regression (quadratic) equations. The alpha diversity (Shannon diversity index), beta diversity, core phyla and genera, and function of the intestinal microbiota were affected by SB supplementation. In addition, the Shannon diversity index and core phyla and genera of the intestinal microbiota were correlated with calf growth-related indices. Overall, SB supplementation in suckling milk improved the growth performance and intestinal microbiota development of dairy calves in a quadratic manner, and regression analysis indicated an optimal supplementation level of 8.78 g/d.