Gliotoxin (GT) is a major and highly potent mycotoxin produced by Aspergillus fumigatus. This study aimed to investigate the effects of GT on mice and its detoxification by clove (Syzygium aromaticum). Aspergillus fumigatus exhibited the highest production of GT on Yeast Extract Sucrose medium, with a concentration of 23.823 μg/kg using HPLC. Mice were divided into four groups: the control group, the GT-treated group (16.75 mg/kg body weight, orally), the clove-treated group (200 mg/kg body weight, orally), and the combined GT and clove-treated group (16.75 mg/kg GT and 200 mg/kg clove, orally). The treatments were administered for three months. This study evaluated the impact of GT on liver function by analyzing blood parameters, liver enzyme activities (alanine aminotransferase [ALT] and aspartate aminotransferase [AST]), and histopathological changes in the liver. The results indicated a significant decrease in hemoglobin concentration, packed cell volume (PCV%), and total red blood cell (RBC) count, leading to normocytic normochromic anemia in the GT-treated group. Additionally, changes in hepatic tissue included severe lymphocytic cuffing around blood vessels, dilated and congested blood vessels, and degeneration and necrosis of hepatocytes. This group also showed a significant increase in white blood cell (WBC) count and elevated ALT and AST activities. Conversely, no significant differences were observed in the groups treated with either the combined GT and clove extract or clove extract alone. Histopathological examination of liver sections supported the serum analysis findings. In conclusion, gliotoxin adversely affected the liver of mice, whereas clove extract effectively mitigated these effects.