The toxicity of 15 flotation reagents (including xanthates, carbamates, thiophosphates, a mercaptobenzthiazole and a frothing reagent) used for concentrating sulfide minerals to five species of mineral-oxidising, moderately thermophilic and acidophilic microorganisms was assessed. The acidophiles tested included four bacteria (a Leptospirillum isolate, Acidimicrobium ferrooxidans, Acidithiobacillus caldus and a Sulfobacillus isolate) and one archaeon (a Ferroplasma isolate). There was wide variation both in terms of the relative toxicities of the different flotation reagents and the sensitivities of the microorganisms tested. In general, the dithiophosphates and the mercaptobenzothiol were the most toxic, while the Leptospirillum and Ferroplasma isolates were the most sensitive of the acidophilic microorganisms. The significance of these findings, in view of the expanding application of ore concentrates bioprocessing, is discussed.