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Objectives were to evaluate effects of supplemental zinc hydroxychloride (HYD; Micronutrients, Indianapolis, IN) on gut permeability, metabolism, and inflammation during feed restriction (FR). Holstein cows (n = 24; 159 ± 8 d in milk; parity 3 ± 0.2) were enrolled in a 2 × 2 factorial design and randomly assigned to 1 of 4 treatments: (1) ad libitum fed (AL) and control diet (ALCON; 75 mg/kg Zn from zinc sulfate; n = 6); (2) ad libitum fed and HYD diet (ALHYD; 75 mg/kg Zn from HYD; n = 6); (3) 40% of ad libitum feed intake and control diet (FRCON; n = 6); or (4) 40% of ad libitum feed intake and HYD diet (FRHYD; n = 6). Prior to study initiation, cows were fed their respective diets for 21 d. The trial consisted of 2 experimental periods (P) during which cows continued to receive their respective dietary treatments. Period 1 (5 d) served as the baseline for P2 (5 d), during which cows were fed ad libitum or restricted to 40% of P1 feed intake. In vivo total-tract permeability was evaluated on d 4 of P1 and on d 2 and 5 of P2, using the paracellular permeability marker chromium (Cr)-EDTA. All cows were euthanized at the end of P2 to assess intestinal architecture. As anticipated, FR cows lost body weight (∼46 kg), entered into calculated negative energy balance (−13.86 Mcal/d), and had decreased milk yield. Circulating glucose, insulin, and glucagon decreased, and nonesterified fatty acids and β-hydroxybutyrate increased in FR relative to AL cows. Relative to AL cows, FR increased lipopolysaccharide-binding protein, serum amyloid A (SAA), and haptoglobin (Hp) concentrations (2-, 4-, and 17-fold, respectively); and peak SAA and Hp concentrations were observed on d 5. Circulating SAA and Hp from FRHYD tended to be decreased (47 and 61%, respectively) on d 5 relative to FRCON. Plasma Cr area under the curve increased (32%) in FR treatments on d 2 and tended to be increased (17%) on d 5 of P2 relative to AL treatments. No effects of diet were observed on Cr appearance. Relative to AL cows, FR increased jejunum villus width and decreased jejunum crypt depth and ileum villus height and crypt depth. Relative to FRCON, ileum villus height tended to increase in FRHYD cows. Feed restriction tended to decrease jejunum and ileum mucosal surface area, but the decrease in the ileum was ameliorated by dietary HYD. In summary, FR induced gut hyperpermeability to Cr-EDTA, and feeding HYD appeared to benefit some key metrics of barrier integrity. [ABSTRACT FROM AUTHOR]