Characterization of halophiles in natural MgSO4 salts and laboratory enrichment samples: Astrobiological implications for Mars

The presence of sulfate salts and limited subsurface water (ice) on Mars suggests that any liquid water on Mars today will occur as (magnesium) sulfate-rich brines in regions containing sources of magnesium and sulfur. The Basque Lakes of British Columbia, Canada, represent a hypersaline terrestrial analogue site, which possesses chemical and physical properties similar to those observed on Mars. The Basque Lakes also contain diverse halophilic organisms representing all three Kingdoms of life, growing in surface and near-subsurface environments. Of interest from an astrobiological perspective, crushed magnesium sulfate samples that were analyzed using a modified Lowry protein assay contained biomass in every crystal inspected, with biomass values from 0.078 to 4.21 mg/g; average=0.74±0.7 mg/g. Bacteria and Archaea cells were easily observed even in low-biomass samples using light microscopy, and bacteria trapped within magnesium sulfate crystals were observed using confocal microscopy. Regions within the salt also contained bacterial pigments, e.g., carotenoids, which were separate from the cells, indicating that cell lysis might have occurred during entrapment within the salt matrix. These biosignatures, cells, and any 'soluble' organic constituents were primarily found trapped within fluid inclusions or fluid-filled void spaces between intergrown crystals. Diffuse reflectance infrared Fourier transform spectroscopy (reflectance IR) analysis of enrichment cultures, containing cyanobacteria, Archaea, or dissimilatory sulfate-reducing bacteria, highlighted molecular biosignature features between 550-1650 and 2400-3000 cm. Spectra from natural salts demonstrated that we can detect biomass within salt crystals using the most sensitive biosignatures, which are the 1530-1570 cm, C-N, N-H, -COOH absorptions and the 1030-1050 cm C-OH, C-N, PO bond features. The lowest detection limit for a biosignature absorption feature using reflectance IR was with a natural sample that possessed 0.78 mg/g. In a model cell, i.e., a 0.5 by 1 μm bacillus, this biomass value corresponds to approximately 7.8×10 cells/g. Based on its ability to detect biomass entrapped within natural sulfate salts, reflectance IR may make an effective remote-sensing tool for finding enrichments of organic carbon within outcrops and surficial sedimentary deposits on Mars