Previous agglomerate-scale heap bioleaching studies have outlined the variations in cell numbers of the liquid and attached phases during colonisation of sterilised ore by a pure culture. In this study, a mixed mesophilic culture was used in agglomerate-scale columns containing non-sterilised low-grade copper ore. Over a six - month period, columns were harvested at various intervals to provide snapshots of the metal distribution and the quantity, location, and ecological variations of mineral-oxidizing microbes within the ore bed. The initial colonisation period in this experiment was dissimilar to previous work, as the indigenous community was retained within the ore-bed throughout acid agglomeration. The overall colonisation phase lasted for approximately 1,000 hours until cell concentrations stabilised. In each column, less than 0.05% of the total cells were found in the leachate, 15-20% in the interstitial phase and the remaining ~80% were attached to the mineral surface. Once cell numbers had stabilised, interstitial cell concentrations were approximately 2,000× greater than those in the leachate. This difference persisted for the duration of the experiment. Copper concentrations in the two liquid phases generally decreased over time, but were on average 50× higher in the interstitial phase. Iron concentrations were more stable, but again were 30× higher in the interstitial phase. This demonstrates that that the difference in cell concentration between the leachate and interstitial phases cannot be explained through diffusion gradients within the system as it is much greater than those observed for the dissolved metals. It also shows that the specific environmental conditions of the interstitial and attached cells are very different to those inferred through analysis of leachates alone.
