The biodiversity of acidophilic prokaryotes was determined in three AMD ponds (pH 2.7-6.5) in Turkey (Izmir-Halikoy antimony ore enrichment plant) using 16S rRNA cloning and denaturing gradient gel electrophoresis methods. Water samples were taken two times in March 2014 and June 2015. The microbial diversity identified includes species such as Acidiphilium angustum, Acidocella sp., Ferroplasma acidiphilum, Acidithiobacillus ferriphilus, Acidithiobacillus ferrivorans, Acidiphilium rubrum, Thiomonas sp., Acidiphilium multivorum, Acidiphilium cryptum, Ferrovum myxofaciens, Acidocella aluminiidurans with the used techniques. In addition to, it has been determined that biodiversity is variable in the operating mine pools. Aciditihobacillus ferriphilus, Acidiphilium angustum, and Acidiphilium rubrum are new records for Turkey.