ALBERT

Description: Physical evidence of life (physical biomarkers) from the deposits of carbonate hot springs were documented at the scale of microorganisms--submillimeter to submicrometer. The four moderate-temperature (57 to 72 degrees C), neutral pH springs reported on in this study, support diverse communities of bacteria adapted to specific physical and chemical conditions. Some of the microbes coexist with travertine deposits in endolithic communities. In other cases, the microbes are rapidly coated and destroyed by precipitates but leave distinctive mineral fabrics. Some microbes adapted to carbonate hot springs produce an extracellular polymeric substance which forms a three-dimensional matrix with living cells and cell remains, known as a biofilm. Silicon and iron oxides often coat the biofilm, leading to long-term preservation. Submicrometer mineralized spheres composed of calcium fluoride or silica are common in carbonate hot spring deposits. Sphere formation is biologically mediated, but the spheres themselves are apparently not fossils or microbes. Additionally, some microbes selectively weather mineral surfaces in distinctive patterns. Hot spring deposits have been cited as prime locations for exobiological exploration of Mars. The presence of preserved microscopic physical biomarkers at all four sites supports a strategy of searching for evidence of life in hot spring deposits on Mars.

Description: Rock, air and service water samples were collected for microbial analyses from 3.2 kilometers depth in a working Au mine in the Witwatersrand basin, South Africa. The approx. 1 meter wide mined zone was comprised of a carbonaceous, quartz, sulfide, uraninite and Au bearing layer, called the Carbon Leader, sandwiched by quartzite and conglomerates. The microbial community in the service water was dominated by mesophilic aerobic and anaerobic, alpha, beta, and gamma-Proteobacteria with a total biomass concentration approx. 10(exp 4) cells/ml, whereas, that of the mine air was dominated by members of the Chlorobi and Bacteroidetes groups and a fungal component. The microorganisms in the Carbon Leader were predominantly mesophilic, aerobic heterotrophic, nitrate reducing and methylotrophic, beta and gamma-Proteobacteria that were more closely related to service water microorganisms rather than air microbes. Rhodamine WT dye and fluorescent microspheres employed as contaminant tracers, however, indicated that service water contamination of most of the rock samples was 〈 0.01% during acquisition. The microbial contaminants most likely originated from the service water, infiltrated the low permeability rock through and accumulated within mining-induced fractures where they survived for several days prior to being mined. Combined PLFA and terminal restriction fragment length profile (T-RFLP) analyses suggest that the maximum concentration of indigenous microorganisms in the Carbon Leader was 〈 10(exp 2) cells/g. PLFA, (35)S autoradiography and enrichments suggest that the adjacent quartzite was less contaminated and contained approx. 10(exp 3) cells/gram of a thermophilic, sulfate reducing bacteria, SRB, some of whom are delta Proteobacteria. Pore water and rock geochemical analyses suggest that these SRB's may have been sustained by sulfate diffusing from the adjacent U-rich, Carbon Leader where it was formed by radiolysis of sulfide.

Description: Rock, air and service water samples were collected for microbial analyses from 3.2 kilometers depth in a working Au mine in the Witwatersrand basin, South Africa. The approx. 1 meter wide mined zone was comprised of a carbonaceous, quartz, sulfide, uraninite and Au bearing layer, called the Carbon Leader, sandwiched by quartzite and conglomerates. The microbial community in the service water was dominated by mesophilic aerobic and anaerobic, alpha, beta and gamma-Proteobacteria with a total biomass concentration approx. l0(exp 4) cells/ ml, whereas, that of the mine air was dominated by members of the Chlorobi and Bacteroidetes groups and a fungal component. The microorganisms in the Carbon Leader were predominantly mesophilic, aerobic heterotrophic, nitrate reducing and methylotrophic, beta and gamma - Proteobacteria that were more closely related to service water microorganisms rather than air microbes. Rhodamine WT dye and fluorescent microspheres employed as contaminant tracers, however, indicated that service water contamination of most of the rock samples was less that 0.01% during acquisition. The microbial contaminants most likely originated from the service water, infiltrated the low permeability rock through and accumulated within mining-induced fractures where they survived for several days prior to being mined. Combined PLFA and terminal restriction fragment length profile (T-RFLP) analyses suggest that the maximum concentration of indigenous microorganisms in the Carbon Leader was less than lo(exp 2) cells/ g. PLFA, S-35 autoradiography and enrichments suggest that the adjacent quartzite was less contaminated and contained -10(exp 3) cells/gram of a thermophilic, sulfate reducing bacteria, SRB, some of who are delta Proteobacteria. Pore water and rock geochemical analyses suggest that these SRB's may have been sustained by sulfate diffusing from the adjacent U-rich, Carbon Leader where it was formed by radiolysis of sulfide.

Description: Previous studies of the subsurface biosphere have deduced average cellular doubling times of hundreds to thousands of years based upon geochemical models. We have directly constrained the in situ average cellular protein turnover or doubling times for metabolically active micro-organisms based on cellular amino acid abundances, D/L values of cellular aspartic acid, and the in vivo aspartic acid racemization rate. Application of this method to planktonic microbial communities collected from deep fractures in South Africa yielded maximum cellular amino acid turnover times of approximately 89 years for 1 km depth and 27 C and 1-2 years for 3 km depth and 54 C. The latter turnover times are much shorter than previously estimated cellular turnover times based upon geochemical arguments. The aspartic acid racemization rate at higher temperatures yields cellular protein doubling times that are consistent with the survival times of hyperthermophilic strains and predicts that at temperatures of 85 C, cells must replace proteins every couple of days to maintain enzymatic activity. Such a high maintenance requirement may be the principal limit on the abundance of living micro-organisms in the deep, hot subsurface biosphere, as well as a potential limit on their activity. The measurement of the D/L of aspartic acid in biological samples is a potentially powerful tool for deep, fractured continental and oceanic crustal settings where geochemical models of carbon turnover times are poorly constrained. Experimental observations on the racemization rates of aspartic acid in living thermophiles and hyperthermophiles could test this hypothesis. The development of corrections for cell wall peptides and spores will be required, however, to improve the accuracy of these estimates for environmental samples.