The study of microbial sulfide oxidation and oxidation rates of tailings requires the use of sterile control samples in laboratory experiments for comparison purposes. The need for sterile tailings sand is also apparent in the study of microbial communities and processes. This study compares six different methods with regard to sterilisation efficiency and investigates chemical and mineralogical changes in the samples as the result of sample treatment. In addition, the effect of sterilisation on mineral weathering rates is also studied. The efficiency of the different methods with regard to sterility is estimated by most probable numbers (MPN) of both sulfur and iron oxidising bacteria. The sterilisation methods that were compared are: washing with sterile water, washing with ethanol, applying antibiotics, repeated heating to 70°C, autoclaving, and γ-irradiation. There are several possible drawbacks with the different methods, including the risk of an applied chemical acting as a substrate for micro-organisms in the system, and the risk of altering the tailings. The results show a substantial difference in sterilisation efficiency between the treatments. For example, autoclaving, repeated heating, ethanol and irradiation efficiently kill micro-organisms, rendering an undetectable amount of bacteria after one month of incubation, whilst washing with water exhibits virtually no microbiological effect, with a bacterial content comparable to the control.
The use of antibiotics for sterilisation ranks intermediate: the lag-phase was twice as long in the antibiotics treatment compared with the controls. X-ray diffraction, optical and electron microscopy, Mössbauer spectroscopy, and mineral magnetism indicate no significant change in the bulk mineralogical composition. However, X-ray photoelectron spectroscopy analyses show that with some of the treatments, sulfides are partially oxidised, as indicated by the formation of sulfate and partially – oxidised sulfur species at the tailings surfaces. Aerated batch weathering experiments performed on sterilised tailings samples and untreated controls indicated that while the initial rates of element (S, Fe, Cu, Zn, Mg, Na, K, Ca) release may differ between the samples, over the course of ~100 days of experiment at pH 2 and room temperature, the rates declined toward a similar value for all samples.
To conclude, there does not appear to be one perfect method for tailings sterilisation. Autoclaving and heating damage the mineral grain structure and oxidise the sulfide surfaces. Ethanol, antibiotics and rinsing greatly alter the pore water chemistry through the washing procedure. In addition, ethanol and denatured antibiotics might serve as a substrate and also, in the case of antibiotics, adhere to mineral surfaces thus obstructing chemical reactions. Irradiation seems to be the preferable method on an overall basis, although the formation of surface Fe3+ was suggested by Mössbauer spectroscopy.
2003. 1013- p.
Sixth International Conference on Acid Rock Drainage (ICARD), Cairns, Australia, July 14 – 17, 2003