In the bioleaching - solvent extraction - electrowinning practices for copper extraction, the solvent extraction process is used to enrich the copper in the leachate to a high lever to make the sequent electrowinning process possible. O/W type of interfacial emulsion always forms at the interface between organic phase and aqueous phase in the solvent extraction practices. Diversified microorganisms were discovered by microscope growing in the interface emulsion of copper solvent extraction. The diversity of them was analyzed by using 16s rRNA gene sequencing. It was indicated that there exist seven categories of bacteria in the interface emulsion, among which common bioleaching bacteria, such as Leptospirillum sp.,Acidithiobacillus ferrooxidans, Metallibacterium sp., are included. The dominant strains are Leptospirillum sp. E4-L9 and Acidithiobacillus ferrooxidans ATCC 23270, which account up to 40.48 % and 38.1 %, respectively. It was uncovered that bioleaching bacteria can be intercepted by interfacial emulsion of solvent extraction to make their population in the raffinate lower and the efficiency of bioleaching decrease consequently.