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This study was designed to compare one-step, two-step and spent medium bioleaching of spent catalyst by adapted Aspergillus niger in batch cultures. Aspergillus niger, which was adapted to heavy metal ions, Ni, Mo, Fe, and W, was grown in medium containing up to 5% (w/v) of spent catalyst. The main lixiviant in bioleaching was gluconic acid, which was produced at all pulp densities in the one-step bioleaching process. Gluconic acid was also produced in the two-step bioleaching process when the spent catalyst was present at pulp densities greater than 1% (w/v). In the spent medium leaching, however, the primary agent was citric acid. The pulp density of the spent catalyst was varied, and this resulted in different amounts of solubilized metals. A total of 3% (w/v) spent catalyst generally produced maximum extraction yields in the one-step bioleaching process; the amounts of leached metals were 100% of W …