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Lake Vostok, East Antarctica

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Lake Vostok, East Antarctica Новости с озера Восток в Антарктиде С.А. Булат (ФГБУ «ПИЯФ», Гатчина) indescribable Elusive microbial life at the uppermost water layer in subglacial Lake Vostok, East Antarctica Sergey Bulat* CryoAstrobiology group/labo FSBI PNPI, NRC Kurchatov Institute Leningrad region, Gatchina, RU *Russian-French GDRI consortium RUB from RFBR, RossHydroMeth GDRI consortium • PNPI FSBI, St Petersburg, RU Main BIO – Sergey Bulat - PI • AARI FSBI, St Petersburg, RU – Vladimir Lipenkov - PI – Valery Lukin – logistics INMI, Moscow • VNIIOceangeologia, St Petersburg, RU – German Leitchenkov LTHE, FR • LGGE CNRS-UJF, Grenoble, FR Jean Martins – Jean Robert Petit - PI – Michel Legrand (DOC) Culturing • Université de Bretagne Occidentale, Brest, FR Microscopy – Mohamed Jebbar • Station Biologique de Roscoff, Roscoff, FR Cell counts – Dominique Marie • Ecologie Microbienne UCB-Lyon I, Lyon, FR Repetition – Philippe Normand Culturing Bulat13 mars-14 FKS2013 Life around us? Searching for extraterrestrial life is one of the aims of astrobiology, and there are several potentially habitable worlds in the Solar System, in addition to Earth. Two of Jupiter’s moons (Ganymede and Europa) and two of Saturn’s moons (Titan and Enceladus) harbour liquid-water oceans. And Mars probably had vast liquid-water environments during much of its early history. Lazcano 2012 Nature 488: 160-1 Bulat13 mars-14 FKS2013 Pinheiro et al 2012 Science 336_341-344 Six new xeno-nucleic acids (XNAs) capable for Darwinian evolution and folding into defined structures Anhydrohexitol nucleic acid, or HNA amongst others: CeNA, LNA, ANA, FANA and TNA Working on Earth-known microbial DNA/RNA 4 primary nucleotides based on: cytosine, guanine, adenine and thymine - C, G, A, T GenBank sequence database “Dust-free” room at LGGE – class 10,000 Fine snow/ice dust particles et DNA analyses DNA/RNA BioCap hood class 100 Columns Centriprep Plus-70 (ml vol.) Biomax membrane 5 kD 135 bp DNA (45.9 nm) Bulat13 mars-14 FKS2013 Number of microbial species (red) Number of microbial genes (blue) Microbial inhabitants outnumber our body’s own cells by about ten to one Relman 2012 Nature 486_194-5 Vostok ice for Biology REQUIREMENTS: • Comprehensive Biological controls – Sham/mock DNA extraction – Negative PCR – Ice core wash water – Lab Environment (dust) – Vostok drill fluid – Outer-core (optionally) 235 seqs Fev 2013 Contaminant Bulat13 mars-14 FKS2013 databases Vostok ice for Biology REQUIREMENTS: • Authentication of findings by ecology (environmental conditions) “Everything is everywhere, but, the environment selects” – Baas Becking, 1934 Bulat13 mars-14 FKS2013 Clean cold and Gatchina – PNPI? DNA extraction certified clean laboratory block Primary PCR No PCR Dubna – JINR? products A framework for biological studies adopted for Lake Vostok Bulat13 mars-14 PNPI NRC Kurchatov Inst Gatchina clean et cold rooms astrobiology laboratory Bulat13 mars-14 FKS2013 Vostok station (+3488m) (78°S, 106°E) (since 1957) 67 kPa – 67% oxygen 1260 km from the coast Absolute minimum -89.2°C Average ann. Temp. -55.1°C Lake Vostok (-3663m) RADARSAT, CSA Vostok SURFACE SNOW Vostok Surface snow up to 4 Kyr (0 – 130m ) Below 3m the snow is pristine Very clean ‘life-hostile’ environ NO FREE (only BOUND) WATER! • Low accumulation – 2.1-2.4g /cm2/a • Low Aeolian dust input (6-6.5 cm/a – snow / needles!) – 2 μm mode; 10-15 ppb (summer time) • Harsh irradiation (UV-B, C etc.) (‘ozone hole’ – Aug- Nov) – + Water-Oxygen free radicals – Oxidized organics (photochemical reactions at ice-air interface) • Low freezing temperatures – below -36oC (-55.1oC ann.) Bulat13 mars-14 FKS2013 as of September 11, 2012 Total ozone over the Antarctic 19 million square kilometers Bulat13 mars-14 FKS2013 NASA Ozone Watch May – August at Vostok Antarctic polar vortex effect 28% – 0% oxygen 10 – 50 km -57 – 0oC Bulat13 mars-14 www.ldeo.columbia.edu Vostok MEGA- et Colossal- snow 2011 Snow drifts by wind for max. 10 km distance CS11 (78,4722S, 106,7796E) (VostokCollossalSnow2011 (RAE 56) 220L ОК! Trouble! Vostok Colossal-snow 2011 Bulat13 mars-14 FKS2013 Courtesy of A. Ekaykin et al (RAE56) • 216km from coastal Progress – PMS11-1 – Crate 23L 8.6L melt PMS11-1 7034.925S, 8124.103E H 2376m • 29km from coastal Progress – PMS11-2 – Crate 23L 7.6L melt PMS11-2 6941.081S, 7633.621E H 865m Vostok MEGA-snow 2011 Метод Electrical Sensing Zone (Coulter counter, Beckman, США) Vostok MEGA- et Colossal- snow 2011 - RESULTS Dust particle counts Sample N/g M/g (ppb) Masse(0.8-2) Masse (2.2-5 ) Mass >5 Vivendi ELGA-26 1347 7.63 2.79 4.30 0.00 Ultra-pure water 577 1.45 1.45 0.00 0.00 3549 9.2 7.28 1.17 0.00 PMS11-1 4821 12.14 9.09 2.27 0.00 216 km Progress 5375 16.19 10.45 4.11 0.00 PMS11-2 4933 14.6 10.22 3.29 0.00 29 km Progress 4241 11.8 8.80 2.00 0.00 3811 10.7 7.90 1.86 0.00 CS11-15 Vostok 3643 11.94 7.52 3.41 0.00 2.8Bulat13-3.0 m marsdeep-14 3754 13.13FKS2013 7.93 4.07 0.00 CS11-15 – light microscopy OLYMPUS BH-2 Overall view (x500) Vostok MEGA- et Colossal- snow 2011 - RESULTS Sample Bacteria / ml Cell counts Roscoff sea water 30961 PMS11-1 - 216 km Progress 45 PMS11-2 - 29 km Progress 0 CS11-15 Vostok - 2.8-3.0 m deep 0 Bulat13 mars-14 FKS2013 Vostok MEGA- et Colossal- snow 2011 - RESULTS PCR - phylotypes Sample Bacteria / ml PMS11-1 - 216 km Progress 45 V4-v8 16S rDNA amplicon sequencing HA-Pseudomonas sp (alpha-proteo) - 80% 99% Ewingella americana (gamma-proteo, Enterobact.) Clinical specimens including wound, Bradyrhizobium sp(elkani) (alpha-proteo) sputum, urine, stool, blood etc. Sources of infection – domestic air conditioning units, ice baths etc. Vostok Contaminant Library Bulat13 mars-14 FKS2013 Human Normal Microbiome Pre CONCLUSION • Vostok [even 29km from coast!] SURFACE snow contains a few(?) cells / ml - NO CELL populations [lifeless] Background contamination? Bulat13 mars-14 FKS2013 Vostok SURFACE solar RADIATION (~1.6 x 105 cells) FKS2013 Summer (January) 2008: exposure during one month BIO-EXPOSURE 2011 Bulat13 mars-14 FKS2013 BIO-EXPOSURE 2011 (~107 cells/ml) RESULTS (only SW2-09 gDNA) No exposure at Vostok (9 days – each day) No signals (v4-v8890bp, v3-v5590bp, v6170bp reg. 16S rDNA) even in one[1st] day! Bulat13 mars-14 FKS2013 CONCLUSION Intensive solar (UV-B?) radiation as effective cell/DNA ‘killing’ factor? Arctic et Antarctic PERMAFROST bioExposure trial in a progress Vostok ICE CORE Towards the subglacial Lake Vostok SALE Subglacial Antarctic Lake Environments - 379 Wright et Siegert , 2012 Lake Vostok known since 1994 Published in Nature (Kapitsa et al., 1996) RADARSAT, CSA Friendly environment? FREE (LIQUID) WATER! • Deeply ice buried (in dark) – 4 km • High pressure – 337-377 bar • Permanently BUT not very cold – -2.65°C • Likely oxygen supersaturated - 800 mg/L (upper bound 700-1300 mg/L) Bulat13 mars-14 FKS2013 OXYGEN in lake water <0.001 ) l / 700-1300 g 700-1300 m ( <0.001 2 O d e v l 175 o s s i 70 D 27 O2 0 1.5 4 30-50 Water cycles Time (t/tR) FKS2013 No life ISOLATED May host life Bulat13 mars-14 RADARSAT, CSA Lake Vostok settings • Huge subglacial lake (Masolov et al., SCAR OSC2008) – 275 x 65 km in size; 15 500 km2 area; 6 100 km3 water – Two main basins with the ridge between 150-200 m deep – Average depth - 300 m; max depth – ~1650 m (south basin) – Water renewal – 80-100 Kyr 2.0-0.8 Ma years old • Buried beneath 2.0-0.8 Myr old 3750-4200 m thick ice sheet • Isolated from surface biota for >14 Ma – Age of water (melted ice) - ~1 Ma • No hydrological links to other lakes Bulat13 mars-14 FKS2013 Vostok ice core 19.02.1990 (78°S, 106°E) North M South F Восток 5Г Meteoric ice ~4km 3650m Ледниковый лед from coast 0.8 Ma 2.0 Ma – 3538m 4200 м 3750 м3623 м Accretion Уровень м оря Melting zone Gases Lake water body 680m Озерный лед Вода Accretion (lake) ice FKS2013 Accr.Фундамент rate – <10 mm/a 300 250 200 150 100 50 0 Расстояние, км -42m ~345m 30m left! 3720 m - Jan 21, 2011 Enter! 3769.3 m – Fev 05, 2012 Bulat13 mars-14 FKS2013 Bulat13 mars-14 FKS2013 Lake water frozen Bulat13 mars-14 FKS2013 We believe Whatever we find It’s going to be special stuff, unique stuff, life forms we’ve never encountered before Accretion ice BIO features Accretion GAS DOC Major ICE NOcontents* GAS NO(ppbC) DOC ions* TYPE 1 Low 10 ± 9 Mg2+ Ca2+ (10-2) SO 2- (S2-) Sediments 4 + CO2, CH4, O2 (no NH4 , nitrate) TYPE 2 Very low 5.6 ± 4.6 - (10-3) (no NH +, Very clean 4 nitrate) (no O2, CH4) Conditions Low aerobic Ultra- No e-donors? Anaerobic Oligotrophic (Hydrogen?) No heterotrophs Many oligotrophic lakes Bulat13 mars-14 FKS2013 have DOC's in the range of 100 to 500 ppb 3608BK - Big ‘Kamina’ Mono-crystalline ice 6 x 8.5 x 15 (mm) 3608m RADARSAT, CSA Microbes in lake water ‘Oxygenophilic’(?) Earth-unknown ! chemolithoautotrophic piezophilic psychrophiles Superoxide, hydrogen peroxide, singlet O, hydroxyl radicals etc. vs. carotinoids, catalase, superoxide dismutase etc. Bacterial metabolic pathways Accretion ice type 1 et water E-donor E-acceptor Carbon Process Microbes 2- H2 SO4 CO2 Sulfate reduction Sulfate- reducers (H2S) H2 O2 (in mineral CO2 Hydrogen- Hydrogen- inclusions) oxidation oxidizers H2 CO2 CO2 Methanogenesis Methanogenic archaea (CH4) 2- 3- S O2 (NO ) CO2 Sulfur oxidation Sulfur- 2- S0 (SO4 - acidic) oxidizers In mineral inclusions RECALL: Gas content is very low - 0.001 vs.
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