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A retrospective cohort study was performed with the objective of determining whether the serum concentrations of nonesterified fatty acids (NEFA), β-hydroxybutyric acid (BHBA), or calcium were associated with the risk of culling within 60 d in milk (DIM) in Holstein cows, and to establish thresholds for each metabolite that were predictive of increased culling risk. Data from 5,979 cows in Ontario (Canada) and several US states were obtained from 4 previously reported studies. For each metabolite and each of 3 sampling weeks (−1, +1, and +2 relative to calving), an optimal threshold was calculated based on having the maximum combined sensitivity (Se) and specificity (Sp) and used to categorize the serum concentrations into high and low risk groups. Logistic regression models were built for each metabolite and each week of sampling, as well as considering together all metabolites in wk −1 and wk +1 relative to …