Sakurako Kimura

The microbial composition of acid streamers (macroscopic biofilms) in acidic, metal-rich waters in two locations (an abandoned copper mine and a chalybeate spa) in north Wales was studied using cultivation-based and biomolecular techniques. Known chemolithotrophic and heterotrophic acidophiles were readily isolated from disrupted streamers, but they accounted for only <1 to 7% of the total microorganisms present. Fluorescent in situ hybridization (FISH) revealed that 80 to 90% of the microbes in both types of streamers were beta-Proteobacteria. Terminal restriction fragment length polymorphism analysis of the streamers suggested that a single bacterial species was dominant in the copper mine streamers, while two distinct bacteria (one of which was identical to the bacterium found in the copper mine streamers) accounted for about 90% of the streamers in the spa water. 16S rRNA gene clone libraries showed that the beta-proteobacterium found in both locations was closely related to a clone detected previously in acid mine drainage in California and that its closest characterized relatives were neutrophilic ammonium oxidizers. Using a modified isolation technique, this bacterium was isolated from the copper mine streamers and shown to be a novel acidophilic autotrophic iron oxidizer. The beta-proteobacterium found only in the spa streamers was closely related to the neutrophilic iron oxidizer Gallionella ferruginea. FISH analysis using oligonucleotide probes that targeted the two beta-proteobacteria confirmed that the biodiversity of the streamers in both locations was very limited. The microbial compositions of the acid streamers found at the two north Wales sites are very different from the microbial compositions of the previously described acid streamers found at Iron Mountain, California, and the Rio Tinto, Spain.

The geochemical dynamics and composition of microbial communities within a low-temperature (≈ 8.5°C), long-abandoned (> 90 years) underground pyrite mine (Cae Coch, located in north Wales) were investigated. Surface water percolating through fractures in the residual pyrite ore body that forms the roof of the mine becomes extremely acidic and iron-enriched due to microbially accelerated oxidative dissolution of the sulfide mineral. Water droplets on the mine roof were found to host a very limited diversity of exclusively autotrophic microorganisms, dominated by the recently described psychrotolerant iron/sulfur-oxidizing acidophile Acidithiobacillus ferrivorans, and smaller numbers of iron-oxidizing Leptospirillum ferrooxidans. In contrast, flowing water within the mine chamber was colonized with vast macroscopic microbial growths, in the form of acid streamers and microbial stalactites, where the dominant microorganisms were Betaproteobacteria (autotrophic iron oxidizers such as 'Ferrovum myxofaciens' and a bacterium related to Gallionella ferruginea). An isolated pool within the mine showed some similarity (although greater biodiversity) to the roof droplets, and was the only site where archaea were relatively abundant. Bacteria not previously associated with extremely acidic, metal-rich environments (a Sphingomonas sp. and Ralstonia pickettii) were found within the abandoned mine. Data supported the hypothesis that the Cae Coch ecosystem is underpinned by acidophilic, mostly autotrophic, bacteria that use ferrous iron present in the pyrite ore body as their source of energy, with a limited role for sulfur-based autotrophy. Results of this study highlight the importance of novel bacterial species (At. ferrivorans and acidophilic iron-oxidizing Betaproteobacteria) in mediating mineral oxidation and redox transformations of iron in acidic, low-temperature environments.