ALBERT

Description: The Licancabur volcano (6017 m) hosts the highest and one of the least explored lakes in the world in its summit crater. It is located 22 deg.50 min. South / 67 deg.53 min. West at the boundary of Chile and Bolivia in the High-Andes. In a freezing environment, the lake located in volcano-tectonic environment combines low-oxygen, low atmospheric pressure due to altitude, and high-UV radiation (see table). However, its bottom water temperature remains above 0 C year-round. These conditions make Licancabur a unique analog to Martian paleolakes considered high-priority sites for the search for life on Mars.

Description: Between October 16th and November 9th 2002, the first NASA Ames DDF Licancabur multidisciplinary expedition initiated the investigation of the biology and environment for life in the highest lake on Earth located at the summit of the Licancabur volcano (6017 m/20,056 ft) at the boundary of Chile and Bolivia. The low oxygen, low atmospheric pressure, high-UV radiation, average temperature, volcano-tectonic and hydrothermal environment make the site a close analog to Martian paleolakes 3.5 billion years ago. The overall goal of the project is to understand through a series of high altitude scientific expeditions what strategies life is using to defend itself against killer-level UV radiation and environmental extreme conditions at this altitude. Several other lakes are located at 4300 m at the foot of the Licancabur volcano (hereafter named laguna Blanca and Laguna Verde). They were also investigated using identical experiments and methods as for the summit lake in order to compare the results and better understand the evolution of survival strategies at transitioning elevations. The lagunas are geothermally heated and many springs provide water at various temperatures. Sources of heat are also suspected for the summit lake as its surface water temperature was measured during the successful ascent at +6 C in a -9 C ambient crater environment (with a wind chill factor of -25 C with a wind blowing almost constantly). Results of this project are expected to provide critical keys to help searching and identifying potential sites for life (extant/extinct) on Mars and developing instruments, experiments and technologies for future missions.

Description: We are currently investigating the biological population present in the highest and least explored perennial lakes on earth in the Bolivian and Chilean Andes, including several volcanic crater lakes of more than 6000 m elevation, in combination of microbiological and molecular biological methods. Our samples were collected in saline lakes of the Laguna Blanca Laguna Verde area in the Bolivian Altiplano and in the Licancabur volcano crater (27 deg. 47 min S/67 deg. 47 min. W) in the ongoing project studying high altitude lakes. The main goal of the project is to look for analogies with Martian paleolakes. These Bolivian lakes can be described as Andean lakes following the classification of Chong. We have attempted to isolate pure cultures and phylogenetically characterize prokaryotes that grew under laboratory conditions. Sediment samples taken from the Licancabur crater lake (LC), Laguna Verde (LV), and Laguna Blanca (LB) were analyzed and cultured using enriched liquid media under both aerobic and anaerobic conditions. All cultures were incubated at room temperature (15 to 20 C) and under light exposure. For the reported isolates, 36 hours incubation were necessary for reaching optimal optical densities to consider them viable cultures. Ten serial dilutions starting from 1% inoculum were required to obtain a suitable enriched cell culture to transfer into solid media. Cultures on solid medium were necessary to verify the formation of colonies in order to isolate pure cultures. Different solid media were prepared using several combinations of both trace minerals and carbohydrates sources in order to fit their nutrient requirements. The microorganisms formed individual colonies on solid media enriched with tryptone, yeast extract and sodium chloride. Cells morphology was studied by optical and electronic microscopy. Rodshape morphologies were observed in most cases. Total bacterial genomic DNA was isolated from 50 ml late-exponential phase culture by using the CTAB miniprep protocol. The 16S rRNA genes were amplified by PCR using both Bacteria- and Archaeauniversal primer sets: 27f and 1492r, 21f and 1492r respectively. Sequences of 16S rRNA gene were determined and initially compared with reference sequences contained in the EMBL nucleotide sequence database by using the BLAST program and were subsequently aligned with 16S rRNA reference sequences in the ARB package (http://www.mikro.biologie.tu-muenchen.de). Aligned sequences were inserted within a stable phylogenetic tree by using the ARB parsimony tool. In this work we report the morphology and phylogenetic characterization of two isolates belonged to Laguna Blanca sediments.

Description: We present data and results from an ongoing project of astrobiological high-altitude expeditions investigating the highest and least explored perennial lakes on Earth in the Bolivian and Chilean Andes, including several volcanic crater lakes nearing and beyond 6,000 m in elevation. In the next five years, they will provide the first integrated long-term astrobiological characterization and monitoring of lacustrine environments and their biology for such altitude. These extreme lakes are natural laboratories. They provide the field data missing beyond 4,000 m to complete our understanding of terrestrial lakes and biota. Research on the effects of UV has been performed in lower altitude lakes and models of UV flux over time are being developed. Lakes showing a high content of dissolved organic material (DOM) shield organisms from UV. DOM acts as a natural sunscreen as it influences the water transparency, therefore is a determinant of photic zone depth. In sparsely vegetated alpine areas, lakes are clearer and offer less protection from UV to organisms living in the water. Transparent water and high UV irradiance may maximize the penetration and effect of UV radiation. Shallow-water communities in these lakes are particularly sensitive to UV radiation. The periphyton can live on various susbtrates. While on rocks, it includes immobile species that cannot seek low UV refuges unlike sediment-dwelling periphyton or alpine phytoflagellates which undergo vertical migration. Inhibition of algal photosynthesis by UV radiation has been documented in laboratory and showed that phytoplankton production is reduced by formation of nucleic acid lesions or production of peroxides and free oxygen radicals. of peroxides and free oxygen radicals. Our project is providing the field data that is missing from natural laboratories beyond 4,000 m and will complement the vision of the effects of UV on life and its adaptation modes (or lack thereof).

Description: The phylum Bacteroidetes represents one of the most abundant bacterial groups of marine and freshwater bacterioplankton. We investigated the diversity of Bacteroidetes in water and sediment samples from three evaporitic basins located in the highlands of northern Chile. We used both 16S rRNA gene clone libraries created with targeted Bacteroidetes-specific primers and separation of specifically amplified gene fragments by denaturing gradient gel electrophoresis (DGGE). DGGE analysis revealed a reduced richness of these organisms in samples from Salar de Huasco (two to four DGGE bands) increasing in Salar de Ascotán (two to seven DGGE bands) and Laguna Tebenquiche at Salar de Atacama (four to eight DGGE bands). Cluster analysis (WPGMA) of DGGE bands showed that bands from Salar de Huasco and Salar de Ascotán grouped together and samples from Salar de Atacama formed separate clusters in water and sediment samples, reflecting different Bacteroidetes communities between sites. Most of the sequences analyzed belonged to the family Flavobacteriaceae and clustered with the genera Psychroflexus, Gillisia, Maribacter, Muricauda, Flavobacterium, and Salegentibacter. The most abundant phylotype was highly related to Psychroflexus spp. and was recovered from all three study sites. The similarity of the analyzed sequences with their closest relatives in GenBank was typically 〈97% and notably lower when compared with type strains, demonstrating the unique character of these sequences. Culture efforts will be necessary to get a better description of the diversity of this group in saline evaporitic basins of northern Chile.