Adhesion plays an important role in bacterial dissolution of metal sulfides, since the attached cells initiate the dissolution. In addition, biofilms, forming after bacterial attachment, enhance the dissolution. In this study, interactions between initial adhesion force, attachment behavior and copper recovery were comparatively analyzed for Acidithiobacillus ferrooxidans, Acidithiobacillus thiooxidans, and Leptospirillum ferrooxidans during bioleaching of chalcopyrite. The adhesion forces between bacteria and minerals were measured by atomic force microscopy (AFM). L. ferrooxidans had the largest adhesion force and attached best to chalcopyrite, while A. ferrooxidans exhibited the highest bioleaching of chalcopyrite. The results suggest that the biofilm formation, rather than the initial adhesion, is positively correlated with bioleaching efficiency.