FIELD WORK AT LUPIN MINE, NUNAVAT TERRITORY CANADA MAY 2004 - OCTOBER 2005

Scientists from two NASA Astrobiology Institute teams, IPTAI and MSU, participated in an initial field trip to the Kinross Lupin gold mine in May 2004. Protocols for sampling anaerobic and aerobic eukaryotes will be developed for th e Lupin project by a third Astrobiology team which is directed by Mitch Sogin at the Marine Biological Laboratory, Woods Hole Oc eanographic Institutes. We anticipate scientists from all of these participating in a second field trip to Lupin which is tentatively schedule for fall 2004.

With the assistance of Drs. Ruskeeniemi (Geological Survey of Finland) ) and Monique Hobbs (Ontario Power Generation), brines of widely varying salinity samples were collected from 11 subsurface sites in May 2004. Brines below the permafrost were collected from six drill holes outfitted with valves and pressure gages and located at the 5 to 6 boreholes of varying salinity were collected at levels 1130 and 880 m levels . Dripping water from open fractures in the roof was collected at the 1130 and 250 m levels. Dripping water at the 250 m level is within the current permafrost. Water recirculated within the mine for drilling activities (service water) was sampled from an open drain at the 1130 m level.
Samplings of water, rocks, biofilms, and microbial mats collected during the May 2004 field trip have been distributed to numerous laboratories and the following research activities are in progress:

•  Growth, isolation and characterization of psychrophilic aerobic and anaerobic microorganisms from environmental (i.e. filter or permafrost cores) samples (Bakermans-Michigan State and Amaral-Marine Biological Laboratory). Characterization of isolates would include phylogenetic identity, membrane lipid composition (Pfiffner-Tennessee), temperature tolerance (including freezing tolerance), salt tolerance, pH range, utilization of substrates (i.e. short chain fatty acids and H2) for carbon and energy sources and stable isotope fractionation (Pratt-Indiana and Sherwood-Lollar-Toronto).

•  DNA (both 16SrDNA and functional genes) analyses of environmental (i.e. filter or permafrost cores) samples and flow cytometry of water samples (Onstott-Princeton) to determine biodiversity and biomass of all microorganisms including those noncultivatable.

•  Phospholipid fatty acid (PLFA) analyses of environmental (i.e. filter or cores) samples (Pfiffner-Tennessee) to determine biodiversity, stress level and biomass of cultivable and noncultivable.

•  Sulfur isotope analyses of both sulfide and sulfate species (Pratt-Indiana) will be combined with 35S activity measurements and DSR and APS reductase gene analyses to delineate the microbial S cycle in environmental (i.e. both water and rock core) samples. 35S microautoradiography could be performed on freshly obtained rock/permafrost core to determine the spatial distribution of this activity with respect to mineralogy and physical properties. These results of these analyses would also complement the S and O isotopic analyses of sulfate being performed by Dr. Frape (Univ. of Waterloo) to identify the sources of sulfate.

•  Organic chemistry of environmental (i.e. both water and rock core) samples (Pratt-Indiana) to establish important electron donors and carbon substrates for microbial respiration and growth. The results of these analyses would also complement the inorganic aqueous chemistry being performed by Dr. Frape (Univ. of Waterloo) and Dr. Ruskeeniemi (Geological Survey Finland).

•  Dissolved H2 and CO by residual gas analysis water samples (Onstott-Princeton) to establish abundance of these important electron donors for microbial respiration and growth (CO). These results would also complement the gaseous chemistry analyses being performed by Dr. Frape (Univ. of Waterloo).

•  N-cycle. Concentration and N isotopic composition of dissolved NH4+ (if any) of environmental samples (water and permafrost core) to constrain the origin of N for microbial growth. DNA analyses of functional genes (e.g., NAR, NIR, NOR, NIF, and AMO) utilized by nitrate reducers, N2 fixers and nitrifiers (Onstott-Princeton). These results would also complement the nitrate and N2 isotopic chemistry being performed by Dr. Frape (Univ. of Waterloo).

• 7. Depending upon success of 1 and 2, culturing of non-cultivable microorganisms using microgel technology (Brockman-PNNL and Diversa Corp.)

• 7. Depending upon success of 1, 2 and 8, complete genome sequence of select psychrophilic microorganisms to identify genes associated with environmental adaptations to freezing stress and nutrient deprivation (Hazen-LBL and JGI ).

• 8. Depending upon the success of 1 and 2, perform microbial activity measurements under in situ conditions and determine the S and C isotopic fractionation associated with microbial sulfate reduction and methanogenesis at low temperatures (Pratt-I n diana, Sherwood-Lollar-Toronto, Bakermans-M ichigan State, Pfiffner-Tennessee ). These data can be combined with qPCR of the methanogenic gene (MCR) to assess in situ rates of methanogenesis. The results of these studies would complement ongoing characterization of stable isotopic composition of aqueous, gas and rock pore species by Dr. Frape (Waterloo) of the Univ. of Waterloo .

• 9. Examine the adaptations of microorganisms to low temperature with depth (which corresponds to time of exposure to low temperatures) to understand the evolution of these low temperature adaptive traits (Bakermans-M ichigan State). Examine changes in the composition of the microbial community with depth to determine how low temperatures are selecting for and against individuals within the community.

• 10. Ground penetrating radar (GPR) survey at low frequencies using the European Space Agency (ESA) Netlander GPR, a version of which is currently deployed around Mars on ESA Mars Odyssey Orbiter. This GPR device is designed to penetrate ~1 km into Martian subsurface, but has not been tested at all on Earth, let alone in a terrain analogous to Mars like that present at Lupin. The subsurface characterization permits unprecedented ground truthing and will constrain interpretations of the current Mars orbiter (Clifford-Universities Space Research Association).

An important component of this research project is the development of devices and instruments for borehole analyses and fluid management (i.e. packers and filtering), life detection, and of biosignatures and autonomous drilling in permafrost terrain with low water/rock ratios. Another important aspect of coring in permafrost will be the testing of alternative drilling fluids, such as gases, that will minimize contamination, preserve ice and methane clathrates and could be adapted to Martian drilling conditions. Initial time frame for NASA Astrobiology activities at Lupin mine is 3 years.