Chilean copper production has been growing in the last 20 years reaching an annual production of 5,557,000 tons of Cu in 2007. For each ton of copper produced, about 200 tons of sterile and low grade ore and 100 tons of tailings are discharged in the environment. Most of these wastes contain significant amounts of sulphide minerals, mainly pyrite, and once submitted to weathering, may produce acid mine drainage. On the other hand, the high price of copper raised the interest for processing by leaching the low grade ore deposited in large dumps. An important part of these mining wastes and low grade ores is located in the Andes, where the mean temperature is usually ~5°C or less. The rate at which bioleaching reactions occur is directly related to the temperature at which the microorganisms (bacteria and archaea) develop. A temperature decrease causes both a decrease of the rate of the involved chemical reactions and a decrease or inhibition of microbial growth. In this work we present the results of the isolation of microorganisms from an old tailing deposit, exposed at low temperatures (5oC) during most of the winter. The isolated microorganisms initially showed a low capacity to oxidize 3g/L Fe(II) sulfate at pH 1.6, and tetrathionate 0.01 M, with an initial pH 4 both at 5oC. However, after successive cultures, microorganisms showed a slow capacity to oxidize both substrates, as well as the sulphide contained in the tailings samples. The terminal Restriction Fragment Length Polymorphism (tRFLP) of the isolated cells grown in basal medium containing Fe(II) showed a nearly pure culture of Acidithiobacillus ferrooxidans. The present study indicates that, even at very low temperatures, microorganisms play an important role in the generation of acid mine drainage and in the oxidation and leaching of sulphide ores.