Abstract: The effect of Fe2+, NH4+ and glucose on the leaching of nickel-bearing pyrrhotite by Thiobacillus ferrooxidans (T.fs) was investigated. Fe2+, NH4+ amendment obviously enhanced the leaching of Ni, Cu and Co. Certain concentration of glucose in the medium inhabited the dissolution of Ni and Co, increased the leaching of Mg and did not obviously affect the leaching of Cu. Ni was leached from nickel-bearing pyrrhotite by both direct reaction of bacteria with minerals and indirect reaction of Fe3+ with minerals, Fe3+ did not obviously influence the leaching of Cu. The leaching of Mg depended on pH of leaching solution.