Background: Displaced abomasum (DA) is one of the most important metabolic disorders of dairy cattle. In DA, ischaemic damage may occur as a result of impaired perfusion due to abomasal displacement, which may result in gastrointestinal mucosal damage. Objective: Investigation of gastrointestinal tissue damage in cattle with right displacement of the abomasum (RDA) and left displacement of the abomasum (LDA) using intestinal-related biomarkers. Methods: Forty-eight DA (24 LDA, 24 RDA) and 15 healthy Holstein dairy cows were enrolled between March 2021 and July 2022. Serum biomarkers including gamma-enteric smooth muscle actin (ACTG-2), liver-fatty acid binding proteins (L-FABP), platelet activating factor (PAF), trefoil factor-3 (TFF-3), leptin, claudin-3 and interleukin-8 (IL-8) concentrations were measured from venous blood samples. Results: L-FABP concentrations in the LDA group and TFF-3 concentrations in the RDA group were lower than in the control group. The leptin concentration of the RDA group was higher than that of the other groups. There was a negative correlation between lactate, leptin and IL-8 concentrations. There was a negative correlation between lactate and TFF-3, whereas leptin and lactate were positively correlated. Leptin was the more reliable biomarker for discriminating between RDA and LDA cases. Conclusion: Changes in serum L-FABP, TFF-3 and leptin concentrations in cattle with DA may reflect acute intestinal injury and the subsequent repair phase. However, these biomarkers had poor diagnostic performance in discriminating between healthy and cattle with DA, while leptin emerged as the most useful marker in differentiating LDA from RDA cases.