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Biotechnological application of extremophiles
R. AmilsCBMSO and CAB
Gran Sasso, November 2019
extreme environmentsgeophysical constrains
- high temperature: hipertehermophiles
- low temperature: psicrophiles- high ionic strength: halophiles
- high pressure: barophiles- high radiation
(adaptation)
Charles Darwin
Yellowstone
submarine hydrothermalism
Uyuni salt flat, Boliviaradiation + desecation
Volcan Illimani, Bolivia
Acidic hypersaline lake (SW-Australia)
radiation (nuclear plantJapan)
Russian drilling base at Vostok
subsurface
extreme environmentsgeophysical constrains
- high temperature: hipertehermophiles
- low temperature: psicrophiles- high ionic strength: halophiles
- high pressure: barophiles- high radiation
(adaptation)
geomicrobiology of metallic sulfides
pyrite, molibdenite, tungstenite (thiosulfate mec.)FeS2+6Fe3++3H2O → S2O3
2-+7Fe2++6H+
S2O32-+8Fe3++5H2O → 2SO4
2-+8Fe2++10H+
rest of sulfides (polisulfide mec.)8MS+8Fe3++8H+ → 8M2++4H2Sn+8Fe2+ (n≥2)4H2Sn+8Fe3+ → S8
o+8Fe2++8H+
S8o+4H2O (S oxidizers) → SO4
2-+8H+
Bacterias come-meteoritos
role of the microbial activity in the leaching of pyrite
SO42-+ H+
Fe2+
Fe3+
microbialactivity
chemical reaction
A B
isolation is fundamental to study the physiological
properties of microorganisms
Fe(II) + 4 O2 + 4H2O Fe(III) + 8H+
oxidation and reduction of Fe by At. ferrooxidans
Sº + Fe(III) + + 4H2O SO42- + Fe(II) + 8H+
Step 1Step 4
Step 3 Step 2
DGGE
phylogeny of cloned 16SrRNAs of Acidithiobacillus spp.
0.10
Thermithiobacillus tepidarius spp. AF023264
Acidithiobacillus spp. AF339743Acidithiobacillus spp. AF359940
Acidithiobacillus spp. AF023264
Leptospirillum ferrooxidans X86776
At. caldus AB023405
At. caldus Z29975At. caldus AF137369
Group II
Group I
Group IIIAcidithiobacillus spp. AF407402
At. ferrooxidans AF465607
At. ferrooxidans AJ457808At. ferrooxidans AJ278719
Acidithiobacillus spp. AF376020
Tinto 3
T1
At. ferrooxidans AF465604At. ferrooxidans X75268
Acidithiobacillus
DSM612 AJ459802
Cells hybridized with LEP636 probe (Cy3-labeled)specific for L. ferrooxidans
DAPI-stained cells
fluorescence in situ hybridization (FISH)
confocal microscopy of pyrite colonized with Acidithiobacillus ferrooxidans
(CARD-FISH)
phylogeny of acidophilic microorganisms detected in Rio Tinto
Firmicutes
Actinobacteria
OP2
OP9OP8
OP3
OP10
Aquificae
Chloroflexi
Thermomicrobia
Deinicocci
Bacteroidetes/Flavobacteria/
Sphingobacteriaa
Fibrobacteres
Spirochaetes
Fusobacteria
Chlorobia
Planctomycetacia
VerromicrobiaeChlamydiae
Acidobacteria
Cyanobacteria
a-Proteobacteria
e -Proteobacteria
b/g -ProteobacteriaEuryarchaeota
Crenarchaeota
Koraarchaeota
OP1
d -Proteobacteria
0.1
+H2O Fe(OH)3+H+
Fe2O3 ( )
SRBAt. ferrooxidansAt. thiooxidans
At. caldus
CO2
(CH2O)n
Acidiphilium spp.Acidimicrobium spp.Ferromicrobium spp.
S2-
SO42-
At. ferrooxidans
Acidiphilium spp.(CH2O)n
CO2
Anoxic[O2] [O2]
Oxic
Fe2+
At. ferrooxidans
L. ferrooxidansFerroplasma spp.Acidimicrobium spp.Ferromicrobium spp.
Fe3+
geomicrobiological model of the water column of Río Tinto
ecological paradox
Bodo sp.
5
Euglena mutabilis
Stichococcus sp.
Mesotaenium sp.
Dunaliella bardawil
Chl orella sp.
Cyanid iumcaldarium
Oxytricha granuliferaColpidium sp.
Pinnularia sp.
Eolimna m inima
Actinophryis sp.
Rotaria sp.
Chlamidom onas sp.
Vahlkampfiaust iana
Animals
Fungi
ViridiplantaeStramenopiles
Alveolata
Amoebae
Red Algae
Cercozoa
Euglenozoa
acidophilic fungi from Río Tinto
endemic plants : Erica andevalensis
Fe hyperaccumulator plants such as Imperata cylindrica produce
jarosite and Fe oxides in the interior of their tissues
EO2
pHCO2 + CH4
MethanobrevibacterMethanosaeta
Methanosarcina
CO2 + H2O
S2- Fe2+ CO2
Acetate + CO2 +H2
S80
ThermodesulfobiumDesulfotomaculumDesulfosporosinusSyntrophobacter
Desulfobulbus
Acidithiobacillus, Sulfobacillus, Acidiphilium
SO42-
Acidiphilium, Pedobacter, Variovorax, PseudomonasAcidithiobacillus
Sulfobacillus, Alicyclobacillus, Ferroplasma, Leptospirillum,
Ferrimicrobium, Ferrithrix
Desulfurella, Thermoplasma, Acidithiobacillus
Chromatiales
NO3-
N2O/N2
Pseudomonas
Clostridium, Desulfitobacterium Propionibacterium, AcidovoraxLysinobacillus, RummelibacillusPseudomonas, DechloromonasSedimentibacter, Alcaligenes
Clostridium, GeobacterBacilllus, PaenibacillusDelftia, CommamonasPseudochrobactrum
NO3-
N2O/N2
(CH2O)n
Syntrophobacter
Organic acids/Alcohols
Paludibacter, Staphylococcus, ClostridiumPropionibacterium, Propionispora, Bacillus
AcidithiobacillusSulfobacillus
Acidiphilium, Acidobacterium, Sulfobacillus, Ferrimicrobium, Ferrithrix, Alicyclobacillus, Ferroplasma, Geobacter,
Aciditerrimonas, Desulfosporosinus NO3-
Fe3+
GEOMICROBIOLOGICAL MODEL OF TINTO RIVER SEDIMENTS
N2O/N2
Acidophilic bacteriophages from Rio Tinto
Phage ACD-RT1 – a myovirus infecting Acidiphilium sp., both host and phage isolated from Rio Tinto.
GEOmicrobiología
mineral deposits of Fe preserved in sedimentary
rocks
iron minerals on sedimentary rocks
Fe deposits older than 106 years
which is the origin of this peculiar extreme acidic environment?
subsurface microbiology
Existing woreholes
Artesian wells
Devoted drills
! "#$%&'() *+%, *+'- .'
! "#$%/ '(0++%1*+'- .'
$"%2*'
$"%//3/&'
45%/+(6.'
45%/+(7.'
45%// (6.'45%// (7.'
45%/+(6.'
45%//(6.'
45%/&'
45%/ ) '
8/ '8&'
Site 1 Site 2
R1
R2
L1 L2
Core Processing Steps
Cores brought to surface Cores in plastic liners, cut, labeled
Bags filled with N2 Anaerobic chamber
Profiling BH10After
drilling
XRD Analysis
cromatografía iónica, BH10
613 m0Depth
NO3-
NO2-
PO4=
SO4=
ppm
cromatografía iónica BH10, ácidos orgánicos
0
0.1
0.2
01
2
3
100
50
0
0
5
10
0
3
6
ppm
Tartrate
Propionate
Acetate
Formate
Oxalate
613 m0Depth
cromatografía de gases
0
0,2
0,4
0,6
0,8
1
1,2
1,4
BH10
%H2 %CO2 %CH4
Total proteins ans sugars, BH10
100 612
LD300Chip, sample from 538mbs, BH10
1 23
45
6
7
0
10000
20000
30000
40000
1→Sulfobacillus acidophilus
2→Bacterioferritin protein5→DPS protein3→Pyrococcus furiosus
4→NAG-NAM polymers
6→Shewanella gelidimarina
7→Poly-Glutamic acid
1 23 456 7
Chip tp detect 16S rRNA genes
enrichment cultures:-pyrite, Fe and S oxidizes- ferric iron and sulfate reducers-methanogens- methanotrophs- denitrifiers- acetogenic bacteria
isolated bacteria from enrichment cultures
• Tessarococcus profundi (-139m) Tessarococcus lapidicaptus (-284m, -336m))• Shewanella hafniensis (-121m) Desulfivibrio oxamicus (-450m)• Rhodoplanes piscinae (-420m) Pseudomonas stutzeri (-420m)• Rhizobium selenitireducens (-538m) Microbacterium saccharophilum (-284m)• Acetoanaerobium notareae (-45m) Citrobacter amalonaticus (-336m)• Cellulomonas fimi (-450m) Pleomorphomonas oryzae (-63m)• Macelibacteroides fermentans (-63m) Oerskovia turbata (-414m)
Parabacteroides chartae (-450m) Nocardiodes pyridinolyticus (-420m)Propinicimonas paludícola (-414m)
IPBSL CARD-FISH
BH10-414,80
THIO820
BET42aGAM42a
SAMPLE EUB338 I-III EUB338 II ALF968 ACD840 BET42a ACI145 GAM42a THIO820 THIO1 SBR385 DSS658 LGC354a LGC354b SUL228 HGC69a SS_HOL1400 CF319a LF655 CYA361 ARC915 MSSH859 MC1109 MG1200 MEB859
BH10-50,00 - - -
BH10-75,00 - - -
BH10-90,00 + + + - - - - - - - - + - + + + + - - + - - - -
BH10-102,60 - + - - - - - - - - - - - - - + - - - - - + - -
BH10-103,50 + + - - - - - + - - - + - + + + - - - + - + - -
BH10-121,80 - + - - + - - - - + - - - + - - - - - - - - - -
BH10-130,80 - - - - + - - - - - - - - - - - - - - - - - - -
BH10-139,40 + + - - + + - - - + - + - + + - - - - - - - - -
BH10-206,60 + - + - - + - - + + + + - + + + - - + + + - - +
BH10-228,60 + + - - - - - - - - - - - - - + - - - + - - - -
BH10-249,80 + - + - + - + + - - + - - - + - - + + - - - - +
BH10-266,30 + + - - - - - - - - - - - - - - - - - - + - - +
BH10-284,00 + + + + + + + + - + + - - - + + + + - + +/++ - - -
BH10-294,45 + + + - - - + - - - - + - + + + - - - - - - - -
BH10-294,65 - - - - - - - - - - - - - + + - - - - - - - - +
BH10-311,10 + - + - + - - - - - + + - - - - - - - - - - - -
BH10-352,65 + - - - - - - - - - - + - + - - - - - + - - - -
BH10-353,15 - - - - - - - - - - - - - - - - - - + - - - - -
BH10-355,70 + + - - - - - - - - - + + - - + - - - - + - - +
BH10-392,90 + - - - - - - - - - - - - - - - - - + + +/+ - - -
BH10-401,90 - - - - - - - - - - - - - + - - - - + + + - - -
BH10-409,70 - - - + - - + - - - - - - + - - - - - - - - - -
BH10-414,00 + - - - - - - - - - - - - - - - - - - + - - - -
BH10-414,80 + - + - + + + + - - - - - - + - - - - - - - - -
BH10-415,30 - - - - - - - - - - - - - - - - - - - - - + + -
BH10-415,97 - - - - - - - - - - - - + - - - - ? - - - - - -
BH10-416,55 - + - - - - - - - - - - - + - - - - - - - - - +
BH10-420,00 + + - - - - - - - - - - - - + + - - + - + - - -
BH10-426,15 + - - + - - + - - + - - - - + - - - - + - - - -
BH10-450,30 + + + - + - + - - - - + + + - - - - - - - - - -
BH10-468,80 + + - - - - - - - + - - - + - - - - - - - + - -
BH10-477,45 + + - - - - - - - - - - - + - - - - - - - - - -
BH10-487,20 + + - + - - - - - - + - - + + + - - - - + - - -
BH10-492,60 + + - - - - - - - + - + - + + + + - - - + - - -
BH10-496,75 + + + - + - + - - - - - - + + + - - - + +/- - - -
BH10-519,05 + + - - - - - - - - - - - + - - - - - + + - - +
BH10-520,00 + + - - - - - + - - + + - + - - - - - + - - - -
BH10-544,00 + + - + + - - - - - - - - + - + - - - - + - - -
BH10-568,60 + + + - - - - - - - - - - + + + - - - - - - + +
BH10-607,60 + + - - - - + - - - - - - + + - - - - - - - - +
BH10-612,94 + + - - - - - + - - - + - - + - - - - + +/+ - - -
SONDAS
Fe and S cycles
10μmAcidovora
x
Sulfobacill
us
Acidiphilli
um
SRB
Sred SoxFe2+ Fe3+
Fe2+
Fe3+
10μm
-139.4m
H2S
Seco
ndar
ysu
lfide
s
SO42-Fe2+
CO2
Organic acids
CO2
S2-
SO4
2-
Methanotrophy
S2-
Pseudomonas
Rhodococcus
Actinobacteria
Desulfovibrio
Sulfobacillus
Desulfosporosinus
Methanococcus
Methanosarcina
Fermentation
SO42-
Chemoheterotrophy(Anaerobicrespiration)
S2-
SO42-
Acidiphillium
Shewanella
Desulfovibrio
Sulfobacillus
Desulfosporosinus
Tessaracoccus
Pseudomonas
Fe2+
e- donorsH2
S2FeSMFe2+
NO2-
CH4Organic acids
e- aceptorsCO2SO4
2-
NO3-
Fe3+
Nitratereductio
n SO42-
At. ferrooxidans
S0
Acidovorax
At. ferrooxidans
Organic matter(buried or produced
chemolithoautotrophically)
subsurface biofilms
-355.7m
-420m
-519.1m
LESSONS LEARNED
- It has been detected a high level of diversity and functional activity in the deep subsurface of the IPB (up to -610m) - Iron can be efficiently oxidized in anaerobic conditions- The detected subsurface microbial activities allow to explain the extreme characteristic conditions of Río Tinto- H2 has an important role as a source of energy in the deep subsurface of the IPB (possible origin: water radiolysis, geochemical, biological)- The most important biogeochemical cycles ( C, N, S, Fe) are operative along the different depths of the solid matrix of the IPB - Subsurface biofilms have been detected in situ for the first time. Biofilms seems to be common in the subsurface eventhough it is considered a high energy consuming strategy not recommended for oligotrophic environment like the subsurfaces
and which is the interest of these extreme acidic
environment ?
Biomining (biohydrometallurgy)
FeS2+6Fe3++3H2O → S2O32-+7Fe2++6H+
S2O32-+8Fe3++5H2O → 2SO4
2-+8Fe2++10H+
8MS+8Fe3++8H+ → 8M2++4H2Sn+8Fe2+ (n≥2)4H2Sn+8Fe3+ → S8
o+8Fe2++8H+
Mars exploration
biohydrometallurgy
heap bioleaching
Bio-reactor
the most extreme condition that chemolithotrophic
microorganisms have to deal with is the high concentration
of toxic heavy metals generated by their metabolism
- Fluorescence in situ hybridization (CARD-FISH) is a technology ready to be applied to biohydrometallurgical operations
Cells hybridized with Lep154 probe specific for
L.ferriphilum. Alexa488.
DAPI
L. ferriphilum in MGM cobaltiferrous concentrate tank reactor
0
10
20
30
40
50
60
70
80
90
Feed R1 R2 R3
Continuous Bioleaching reactors
Cells
det
ecte
d (%
)
EUB33 NTR12 GAM42a LEP154 THIO1 SUL1238 FER656+TMP65454
100
endemic plants: Erica andevalensis
future: combination of bio-techniques. Biomining +
specific sequester of metals using acidophilic fungi
acidophilic fungi
• Tabla 2. Eficiencia y especificidad de secuestro de metales pesados• _____________________________________________________•• Aislados concentración de metal secuestro %•• Bahusacala sp. O66 1mM Ag(I) 66• Scytalidium sp. P65 10mM Cd(II) 90• Penicillium sp. I25 200mM Zn(II) 93• Penicillium sp. P34 100mM Cu(II) 35• 10mM As(V) 68• Penicillium sp. V80 100mM Cr(III) 75• Alternaria sp. I14 0.1mM Hg(II) 95
Mars exploration
habitability
Misión MER, crater Endurance,2005
SPIRIT IN GUSEV CRATER
blueberries in Columbia Hills
sulfates exhumed in crater Gusev
silica exhumed (hidrotermal?)
MEX, wáter vapor (SPICAM)
Fe oxides distribution (MEX)
Curiosity
Crater Gale
MSL, arm with instruments
MSL ChemCam,elemental analysis
Mars Express
it can be concluded that on Mars there are sedimentary rocks that were formed in
acidic conditions (acidic lakes or oceans)
terrestrial analogues: - acidic environments
- hydrothermal activities
comparison between MARS and RIO TINTO
MP RTsurf RTss• - hematite ++ ++ +• - jarosite ++ ++ +• - goethite ++ ++ +• - ionic strength ++ ++ ++• - temperature suf low 4-35oC• - temperature subs ? 25-30oC• - methane + - +• - oxygen +/- ++ -• - µorganisms ? ++ +
the actual conditions on the surface of Mars, intens radiation and very oxidant conditions,
do not seems to be the ideal place for life development (mechanisms of protection,
methodological problems). Life on the subsurface has much more possibilities. It is
important to develop a martian drilling mission in a near future.
.
the exploration and characterization of the
Tinto ecosystem is important to understand
the properties of microorganisms that could develop on Mars. Also is a
good bench to test instruments designed to
detect life on Mars…
and to better understand the period in which life appeared
on Earth (Archean)
banded iron formations of Pilbara (Australia)
THANK YOU
isolation in acidic waters from a coal mine
of Thiobacillus
ferrooxidans
terrestrial acidic environments
natural acidic environments:- areas with volcanic activity
SO2 + H2S → S0 + H2O
Yellowstone
natural acidic environments:- metal mining activitiesFeS2 + H2O —> Fe3+ + SO4
2- + H+
in these cases the extreme acidic conditions are promoted by biological activity
Biomining, the future of miningR. Amils
CBMSO and CAB
Bolonia, november 2018
»
isolation in acidic waters from a coal mine of Thiobacillus
ferrooxidans
metabolisms involved
la irrupción de técnicas moleculares al estudio de la ecología microbiana ha sido
una auténtica revolución
DGGE
mesocosms
Cells hybridized with LEP636 probe (Cy3-labeled)specific for L. ferrooxidans
DAPI-stained cells
hibridación in situ con marcadores fluorescentes (FISH)
16S rRNA gene-based oligonucleotide microarray
filogenia de los microorganismos acidófilos detectados en la cuenca del Tinto
Firmicutes
Actinobacteria
OP2
OP9OP8
OP3
OP10
Aquificae
Chloroflexi
Thermomicrobia
Deinicocci
Bacteroidetes/Flavobacteria/
Sphingobacteriaa
Fibrobacteres
Spirochaetes
Fusobacteria
Chlorobia
Planctomycetacia
VerromicrobiaeChlamydiae
Acidobacteria
Cyanobacteria
a-Proteobacteria
e -Proteobacteria
b/g -ProteobacteriaEuryarchaeota
Crenarchaeota
Koraarchaeota
OP1
d -Proteobacteria
0.1
UV radiation protection with Fe3+
radiation protection by ferric iron
01234567
cells x10+5/ml
96 hours incubation
Dunaliella control, 0 hours
Dunaliella control, 96 h
Dunaliella RT media, 5 mWcm-2
Dunaliella RT media, 10 mWcm-2
Dunaliella A media, 5 mWcm-2
Dunaliella A media, 10 mWcm-2
Fe meteorites (irons) chemolithoautotrophy
t0
1 día
3 días
7 días
“irons” as a source of energy
meteorite oxidation by chemolithoautotrophic microorganisms
phyllosilicates can be generated in acidic conditions (Río Tinto)
basic knowledge is needed to improve the efficiency
of bioleaching
Probes used in Petiknas Zn 35ºC, Petiknas Zn 35ºC and Aguablanca 35ºCProbe Targe
tet Sequence (5’ to 3’) (%) FMa Specificity Reference
EUB338 16S GCT GCC TCC CGT AGG AGT 0-35 Bacteria domain Amann, 1990
EUB338-II 16S GCA GCC ACC CGT AGG TGT 0-35 Planctomyces Daims, 1999
EUB338-III 16S GCT GCC ACC CGT AGG TGT 0-35 Verrumicrobia (and others) Daims, 1999
THC642 16S CAT ACT CCA GTC AGC CCG T 35 Acidithiobacillus caldus Bond, 2000
LEP154 16S TTG CCC CCC CTT TCG GAG 35 Leptospirillum ferriphilumGonzález-Toril, 2003
SUL141 16S CGG CCC GAT ATC CCC CAC 35 Sulfobacillus spp. Diez et al. BIOMINE
FER656 16S CGT TTA ACC TCA CCC GAT C 35 Ferroplasma spp. Edwards, 2000
NON338 ----- ACT CCT ACG GGA GGC AGC 35 Negative control Amann, 1990
Cells hybridized with Sul141 probe specific for Sulfobacillus
sp. Alexa488.
DAPI
Tabla 3. Resistencia constitutiva e inducible a metales pesados__________________________________________________
Aislados resistencia constitutiva máximo nivel de resistencia
Cladosporium sp. Y18 < 1mM Cr(III) 400mM Cr(III)Nigrospora sp. V12 1mM Ag(I) 1mM Ag(I)
Penicillium sp. P34 < 1mM Cu(II) 200mM Cu(II)Penicillium sp. Y22 < 1mM Zn(II) 400mM Zn(II)
Tabla 4. Eficiencia del secuestro específico de Cr(III) de Penicillium V80___________________________________________________________
tipo de crecimiento biomasa tiempo de exposición (días) medio secuestro %
crecimiento activo 0.93g (final) 7 YEPD 75.7crecimiento activo 0.8g (final ) 7 50% YEPD 34.9crecimiento activo 0.76g (final) 7 C 39.8fase estacionaria 1g 1 YEPD 1.2células rotas 1g 1 YEPD 0.9
Tabla 5. Comparación de eficiencias de recuperación de V80 utilizando distintos tratamientos
________________________________________________________________número de ciclos [Cr(III)] (mM) inóculo tiempo (días) eficiencia% Cr (moles)
tratamiento #1
un ciclo (A) 100 fresco1/100 7 73.9 73.9dos ciclos (A+B) 26.1 ½ A 2 55.1 87.5tres ciclos (A+B+C) 12.5 ½ B 2 11.1 88.9
volumen total tratado: 100ml
tratamiento #2
un ciclo (M) 100 fresco 1/100 7 79.9 79.9dos ciclos (M+N) 100 ½ A 2 40.2 120.1tres ciclos (M+N+L) 100 ½ B 2 3.1 123.2
volumen total tratado: 300 ml
Themal area of Gunhuver, Iceland
contact/no-contact (direct/indirect)
confocal microscopy of pyrite colonized with Acidithiobacillus ferrooxidans
(CARD-FISH)
Fe content of the core 8,68c
MARTE project, CARD-FISH of sample from core 8,50a
(-107m)
Multianalyte Sample
Incubation with theAb microarray (50-150 m spot diameter)
Addition and incubation with
fluorescent antibodies
Wash
Scanning
Image
Wash
0Antibodies
Quantification of the signal
Immunoprofiling by Sandwich Microarray Immunoassay (SMI)
0Antibodies
C2- Buffer
0Antibodies
Inte
nsity C1- Burnt sample Sample
5 mm
10 mm
800 spots
IPBSL
Nature Reviews Microbiology 6, 339-348
CARD-FISH: Catalyzed reporter deposition Fluorescence In Situ Hybridization
Alexa-488
anaerobic methanogenicconsortium
IPBSL
Sulfolobus acidocaldarius
0,0
10,0
20,0
30,0
40,0
50,0
60,0
70,0
80,0
90,0
100,0
Petiknas Zn 35 Petiknas Zn 45
EUB338+II+III
THC642
LEP154
SUL141
FER656
41,2
25,5
3,9
28,5
62,8
70,9
22,9
1,8
3,9
92,1% o
f m
icro
orga
nism
s
Samples
heap leaching
Tabla 1. Perfiles de resistencia a metales exhibidas por hongos acidófilos___________________________________________________________
Aislados valores máximos de resistencia otras resistencias
Scytalidium sp. O64 1mM Hg(II), 400mM As(V) 200mM Cr(III)Cladosporium sp. I18 400mM Zn(II) y Cr(III) 50mM Cu(II)
Cladosporium sp. P72 400mM As(V) 100mM Zn(II), 200 mM Cr(III)Alternaria sp. I14 1mM Ag(I) ---
Aspergillus sp. P37 --- 50mM Cu(II), As(V) y Cr(III)Aspergillus sp. P51 100mM Cr(III) ---Bahusakala sp. O62 1mM Ag(I) ---
Bahusakala sp. O66 400mM As(V), 1mM Ag(I) y Hg(II) 10mM Ni(II) y Cd(II) Penicillium sp. P54 1mM Ag(I), 400mM As(V) 50mM Cu(II), 100mM Cr(III)
Penicillium sp. V80 200mM Cr(III) ---Penicillium sp. P34 200mM Cu(II), 1mM Ag(I) 50mM As(V), 100mM Cr(III)
Hormonema sp. I12 400mM As(V), 1mM Ag(I) 200mM Cr(III)Hormonema sp. I17 1mM Ag(I) 50mM Zn(II)
Nodulisporium sp. V56 400 mM As(V), 10mM Ni(II) 50mM Cr(III), 10mM Zn(II)Nodulisporium sp. V58 400mM As(V) 100mM Cr(III)Trichoderma viride O6 1mM Ag(I), 10mM Ni(II) y Cd(II) 200mM As(V)
Trichoderma viride CECT 2423 --- ---
EL CAPITAN (MP)
temperature gradient (Endurence)
clauds at Meridiani Planum
Viking I
16S rRNA gene-based oligonucleotide microarray
enrichment cultures:- pyrite, Fe and S oxidizers- Fe and sulfate reducers-methanogens- metanotrophs- denitrifiers- acetogenic bacteria
4H2 + CO2 → CH4 + 2H2O BH2 + SO4
2- + H+→ HS- + 4H2O BFe2+ + H2S→FeS + H2 Q
FeS + H2S → H2 + FeS2 Q FeS2+Fe3++8H2O→2SO4
2-+ Fe2++16H+ BFe2+ + NO3
-→Fe3+ + NO2- B
Cn(H2O)n+Fe3++H2O→CO2+Fe2+4H+ BH2 + HCO3
- + H+→ CH3COO- + H2O BC3H6O3 + SO4
2- → CH3COO- + S2- B
H2O on Mars
MER OPPORTUNITY AT MERIDIANI PLANUM
Crater Eagle
rock outcrop at Eagle crater (MP)
robotic arm with instruments
blueberries, Endurance crater,
SPIRIT AT GUSEV CRATER
blueberries at Columbia Hills
exhumed sulfates at Gusev crater
exhumed silica (hydrothermal?)
MEX, water wapor (SPICAM)
Fe oxides distribution on Mars (MEX)
HRSC-MRO
HRSC-MRO
paleo-ocean (K, Th, Fe) Gamma Ray spectrometer (Mars Odissey)
MEX, phylosilicates (OMEGA)
MEX, H2O-ice in the South Pole (MARSIS)
Phoenix landing site, june 2008
MGS
craterización reciente, MGS
MEX, methane (PFS)
Curiosity
Crater Gale
meteorito de Fe-Ni marciano
MSL, brazo con instrumentación
MSL ChemCam, análisis elemental
se puede concluir que en Marte existen rocas sedimentarias
formadas en condiciones ácidas (lagos o océanos ácidos)
análogos terrestres:
- ambientes ácidos- hidrotermalismo submarino
comparación entre MeridianiPlanum y Río Tinto
MP RTsurf RTss• - hematites ++ ++ +• - jarosita ++ ++ +• - goetita ++ ++ +• - fuerza iónica ++ ++ ++• - T superficie low 4-35oC• - T subsuelo ? 25oC• - metano + - +• - oxígeno +/- ++ -• - µorganismos ? ++ +
Las condiciones actuales de la superficie de Marte, fuerte irradiación UV y condiciones muy oxidantes, no parecen ser el lugar ideal para el desarrollo de
la vida (mecanismos de protección, problemas metodológicos). La vida en el subsuelo tiene
muchas más posibilidades que en la superficie. Es importante diseñar y desarrollar una misión de perforación si queremos detectar vida en Marte
Mars Express
.
La exploración y caracterización del ecosistema del Tinto es importante para
entender las propiedades de los microorganismos que se podrían
desarrollar en Marte. Además es un buen banco de pruebas para probar las
prestaciones de los instrumentos que volarán a Marte en futuras misiones de
exploración…
y para entender mejor el periodo en el que apareció la
vida sobre la Tierra (Archean)
formaciones de hierro bandeado de Pilbara (Australia)
the irruption of molecular biology techniques into microbial ecology has produced an authentic
revolution
• Proteobacteria (Acidithiobacillus, Acidiphilium, Acidiferrobacter, Ferrovum)
• Nitrospira (Leptospirillum)• Firmicutes (Alicyclobacillus, Sulfobacillus)• Actinobacteria (Ferrimicrobium,
Acidimicrobium, Ferritrix)• Archaea (Sulfolobus, Acidianus,
Metallosphaera, Sulfurisphaera, Ferroplasma)
Fungal diversity
Eurotiomycetes
Índice de bootstrap:100%
90-99%80-89%
XRD and Mössbauer spectra
MARTE project, SEM of a sample from core 8,68c (-162m)
MEX: OMEGA
meteorito de Fe-Ni marciano
MARTE project: geomicrobiological exploration of the Iberian Pyritic Belt
subsurface