Compared to prevalent alkaline to neutral hypersaline environments, acidic hypersaline environments have been scarcely
studied. However, they hold interest to many researchers in that these environments have similar geological and
geochemical characteristics as those found in lithified strata on Mars. Fieldwork indicated that Lake Brown, located in
Western Australia, possessed pH values of 3.1-4.5 and salinity between 13.0-23.0%. Water column, groundwater, and
sediment samples were collected from the lake during the austral winter of 2005. These samples were analyzed with
both traditional culture and molecular methods. Modified growth media and minimal media were designed to match the
composition (Cl, Na, Mg, SO4, K, Ca, and Br) of Lake Brown surface and ground waters for the enrichment of
microorganisms. One of the isolates obtained, Brown 1, can grow in media that possesses pH values of 3-7 with optimal
growth at pH 4, salinity that ranged from 5% to saturation with optimal growth at 5% and could grow under
temperatures that ranged from 20°C to 65°C with optimal grow occurring at 37°C. The isolate's optimum growth
conditions are similar to those found in Lake Brown. The isolate is a Gram-negative rod that forms yellow colonies on
17% Phytogel. Its 16S rRNA gene can be amplified with bacterial primers but not with archaeal primers. Its 16S rRNA
gene sequence suggests that the isolate is a gamma proteobacterium. Studies on organisms isolated from environments
such as Lake Brown, an acid hypersaline lake, can provide an opportunity to both expand our knowledge of terrestrial
extremophiles and gain insight on the possible forms of life that might have existed on Mars.
The surface of Mars is exposed to higher levels of solar and galactic cosmic ray irradiation than Earth due to its
very weak magnetic field. Thus, microorganisms that could possibly survive in the shallow subsurface of Mars would
likely be radiotolerant. To better understand microorganisms that might reside in this environment of Mars, a number of
isolates were obtained from the area around a gamma-radiation source, 137Cs, located on the UMR campus. Radiation
sensitivity assays were performed on the isolates as well as on the common bacterium, E. coli. All the organisms tested
were able to withstand exposures up to 20 Gy. The E. coli control did not survive exposures of 200 Gy, while the isolate
designated 1B-1 could. Another isolate, Cont-1, also withstood this exposure. Each of the isolates produced white
growth on solid medium and their cells are rod-shaped. The study of these isolates and similar organisms could enhance
our knowledge of these unique extremophilic bacteria and might provide insight into the microorganisms that could be
present in the shallow subsurface of Mars.
Soap Lake is a haloalkaline lake located in central Washington. This lake is a remnant of the Missoula flood events that
created the landscape of western Montana, the southeastern portion of Washington state, and much of Oregon. It is
15,000 - 20,000 years old, and has maintained a stable meromixis for the last 10,000 years. This carbonate lake is
characterized by a brackish mixolimnion, and a monimolimnion with a salinity of ~14%. The pH of both layers of the
lake is approximately 10. Both layers also have a high concentration of dissolved sulfate, with the mineral mirabilite
(Na2SO4•10H2O) found in the monimolimnion sediments. Sulfide concentrations in the monimolimnion exceed 100 mM.
As part of the mission of the NSF Soap Lake Microbial Observatory, microorganisms involved in the sulfur cycle in this
lake were studied in terms of their diversity and function. High rates of sulfate reduction were measured in both layers of
the lake, with new species of sulfate-reducing bacteria seen in both areas. A particularly novel psychrophilic sulfur
oxidizer was isolated from the monimolimnion. This organism has the ability to induce the formation of mirabilite,
which was assumed to be an abiotically deposited evaporite mineral. This is the first evidence for a biogenic origin of
this mineral. This leads to the possibility that related sulfate minerals, such as those reported on the Mars surface, may
have a biogenic origin.
Recent findings by the NASA's Mars Exploration Rovers and ESA's Mars Express indicate that during an earlier era in the planets' evolution, evaporation of surface water may have left behind vast evaporite deposits1,2,3. This makes the possibility of finding biological material preserved in halite inclusions most intriguing4. The retrieval and characterization of microorganisms from ancient halite crystals5,6 suggests that it might be possible to locate their remains as biomarkers or even living cells from evaporites sampled from extraterrestrial environments. However, before this is possible, techniques for the detection of bacterial cells or biomolecules in halite and other evaporite crystals need further refining. Specifically, contamination must be minimized and quantified during the microbial analysis of such crystals. Aseptic techniques that allow for the direct extraction of fluid brines from micron to millimeter size inclusions significantly reduce the possibility for contamination. However, even with extreme precautions, the possibility for contamination cannot be entirely eliminated, particularly when culture-based methods are employed. We have elicited native fluorescence from a variety of biomolecules, including the aromatic amino acids and nucleic acids, by laser excitation at 248 and 224 nm from haloarchaea and haloarchaea residues trapped in halite. Energy to each sample, (positive control crystals with Halobacteria salinarum and bacteria-free negative control crystals), was 80 microwatts at 224 nm and 25 microwatts at 248 nm. A 30 s exposure of the inclusions within the positive control elicited easily detectable fluorescence while there was no response from the negative control crystals during the same exposure. Analysis of halite crystals sampled from the Waste Isolation Pilot Plant, Carlsbad, New Mexico yielded similar results. To minimize microbial damage from the high-energy 224-248 nm beams and to make the technique more widely available to the scientific community, we have examined the possibility of using a standard epi-fluorescent microscope for similar purposes. We have elicited a native fluorescence response from microscopic eukaryotes in rapid scanning, low magnification mode employing 365 nm excitation and are optimizing the visualization of prokaryotes with this system. Aseptic identification of epifluorescent biosignatures in evaporite inclusions would be of significant utility for planetary protection and preliminary screening protocols during a Mars sample return mission.
Conference Committee Involvement (4)
Instruments, Methods, and Missions for Astrobiology XI
12 August 2008 | San Diego, California, United States
Instruments, Methods, and Missions for Astrobiology X
28 August 2007 | San Diego, California, United States
Instruments, Methods, and Missions for Astrobiology IX
14 August 2006 | San Diego, California, United States
Instruments, Methods, and Missions for Astrobiology IX
31 July 2005 | San Diego, California, United States
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