This paper aimed to study the effect of copper sulfate fertilization (90 kg/hm2) on molybdenum (Mo) poisoning of the Przewalski’s gazelle (P. przewalskii) in the Qinghai Lake basin. Soil, pasture, and the Przewalski’s gazelle’s blood and hair samples were collected from the animal rescue center in this study. The biochemical parameters of the Przewalski’s gazelle’s blood and the mineral contents of the fodder and soil were ascertained. The findings demonstrated that the impacted pastures’ soil and forage had considerably greater Mo levels (p < 0.01) than those of the healthy pastures. Compared to the control group, the experimental gazelles had significantly lower Mo content in the blood and hair, while the impacted gazelles had significantly higher copper and sulfur contents in the blood and hair (p < 0.01). The levels of blood Hb, PCV, MCV, and MCH of the Przewalski’s gazelle in the fertilized group had been extensively greater than those in the control group (p < 0.01). The tiers of blood AST, LDH, CPK, and ALP of the Przewalski’s gazelle in the fertilized group had been extensively greater than those in the control group. The contents of TP, ALB, and GLB in the fertilized group had been substantially decreased than those in the control group. In comparison to the control group, the fertilized group exhibited considerably higher activity of serum GSH-Px, SOD, T-AOC, and CAT and a significantly lower MDA concentration. The results showed that the appropriate application of copper sulfate could significantly increase the copper content in forage, regulate the molybdenumcopper ratio in soil, and thus effectively improve the molybdenum toxicity symptoms of Przewalski’s gazelle, improve the antioxidant ability of the gazelle, and alleviate the molybdenum toxicity condition.