Mitigation strategies are vital in minimizing the health and economic risks associated with dairy cattle exposure to aflatoxin (AF). The objective of this study was to determine the effects of a commercially available aluminosilicate clay in a traditional lactation diet during an AF challenge on inflammatory markers of multiparous lactating Holstein cows. Sixteen multiparous, lactating Holstein cows [BW = 758 ± 76 kg; DIM = 157 ± 43 d] were assigned to 1 of 4 treatments in a replicated 4 × 4 Latin Square design: no adsorbent and no AF challenge (CON), no adsorbent and AF challenge (POS), 113 g of aluminosilicate clay top-dressed on the TMR (adsorbent; PMI Nutritional Additives, Arden Hills, MN) with AF challenge (F4), and 227 g of adsorbent with AF challenge (F8). For each period, blood was sampled on d 14, 18, and 21, and liver was sampled on d 18. Gene expression and histological hepatocyte inflammation were assessed for liver tissue. One orthogonal contrast (POS vs. CON) was used along with the linear and quadratic effects of treatments POS, F4, and F8. Multivariable logistic mixed models considering the binary outcome variable histological hepatocyte inflammation score were constructed. Cows in CON had greater concentrations of serum cholesterol (202 mg/dL; P < 0.001) and plasma superoxide dismutase (2.8 U/mL; P = 0.04) than cows in POS (196 mg/dL and 2.6 U/mL, respectively). Plasma glutamate dehydrogenase increased as concentration of adsorbent in the diet increased (POS = 37.8, F4 = 39.3, F8 = 39.1 U/L; P = 0.05). NFKB1 expression was higher (P = 0.04) in liver of cows in POS (0.78 ± 0.04) compared with cows in CON (0.70 ± 0.04). MTOR expression was higher (P = 0.04) in liver of cows in CON (1.19 ± 0.06) compared with cows in POS (0.96 ± 0.06). When compared with cows in CON, cows in POS had higher (P = 0.05) odds ratio for hepatocyte inflammation (OR = 5.14; 95%CI = 0.97–27.33). When compared with cows in F8, cows in POS tended (P = 0.07) to have higher odds ratio for hepatocyte inflammation (OR = 4.60; 95%CI = 0.90–23.66). In conclusion, AF exposure lead to reduced function and increased inflammation of the liver. Additionally, the adsorbent used helped to decrease hepatocyte inflammation and neither hindered nor improved blood metabolites and liver gene expression.