Glacial ice cores: A model system for developing extraterrestrial decontamination protocols

Evidence gathered from spacecraft orbiting Mars has shown that water ice exists at both poles and may form a large subsurface reservoir at lower latitudes. The recent exploration of the martian surface by unmanned landers and surface rovers, and the planned missions to eventually return samples to Earth have raised concerns regarding both forward and back contamination. Methods to search for life in these icy environments and adequate protocols to prevent contamination can be tested with earthly analogues. Studies of ice cores on Earth have established past climate changes and geological events, both globally and regionally, but only recently have these results been correlated with the biological materials (i.e., plant fragments, seeds, pollen grains, fungal spores, and microorganisms) that are entrapped and preserved within the ice. The inclusion of biology into ice coring research brings with it a whole new approach towards decontamination. Our investigations on ice from the Vostok core (Antarctica) have shown that the outer portion of the cores have up to 3 and 2 orders of magnitude higher bacterial density and dissolved organic carbon (DOC) than the inner portion of the cores, respectively, as a result of drilling and handling. The extreme gradients that exist between the outer and inner portion of these samples make contamination a very relevant aspect of geomicrobiological investigations with ice cores, particularly when the actual numbers of ambient bacterial cells are low. To address this issue and the inherent concern it raises for the integrity of future investigations with ice core materials from terrestrial and extraterrestrial environments, we employed a procedure to monitor the decontamination process in which ice core surfaces are painted with a solution containing a tracer microorganism, plasmid DNA, and fluorescent dye before sampling. Using this approach, a simple and direct method is proposed to verify the authenticity of geomicrobiological results obtained from ice core materials. Our protocol has important implications for the design of life detection experiments on Mars and the decontamination of samples that will eventually be returned to Earth.