Bioleaching as a low cost and environment-friendly process could be a promising option for the enrichment of froth flotation products. Flotation reagents (collectors, frothers, etc.) are effective on the bacteria growth and oxidation activity; however, their impact has not been widely investigated. In this study, the effect of conventional reagents for sulfide flotation; collectors: potassium amylxanthate (KAX), potassium isobutyl-xanthate (KIBX), sodium ethylxanthate (NaEX), potassium isopropyl xanthate (KIPX) and Dithiophosphate (Aero3477), and frothers; pine oil (PO) and methyl isobutyl carbinol (MIBC) in various concentrations have been examined on Acidithiobacillus ferrooxidans activities. The results of this study demonstrate these flotation surfactants may have positive or negative influences on the bio-oxidation, based on their chemical compositions and/or concentrations. In general, the inhabitation effects of collectors would be increased in higher dosages and based on differences between results of various conditioning tests with the control test (without reagent) in different days, this effect could be considered by the following order: for 0.01 g/L: KAX > KIPX > KIBX > Aero3477 > NaEX, 0.1 g/L: NaEX > KIPX > KAX > KIBX > Aero3477, and 1 g/L: NaEX > KIPX > KIBX > KAX > Aero3477, and for frothers: MIBC > PO in all concentrates. These outputs potentially can be used for the selection of flotation surfactants when the flotation products are going to be further processed by bioleaching for the metallurgical extraction.