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D. MARTY, J. GARCIN, C. TAMBURINI, A. BIANCHI. BIODEEP Meeting VIII VENEZIA 13-15 October 2003. BACTERIAL ACTIVITIES. * STRICT ANAEROBIC ACTIVITIES * Methane Production (MP) * Sulfate Reduction (SR). * BACTERIAL PRODUCTION & ACTIVITY * Bacterial Production (BP)

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Wp5 microbial metabolic activities cnrs dr 12 lmm

D. MARTY, J. GARCIN, C. TAMBURINI, A. BIANCHI

BIODEEP Meeting VIII VENEZIA 13-15 October 2003

BACTERIAL ACTIVITIES

  • * STRICT ANAEROBIC ACTIVITIES

    • * Methane Production (MP)

    • * Sulfate Reduction (SR)

* BACTERIAL PRODUCTION & ACTIVITY

* Bacterial Production (BP)

* Glutamate & Glucose Uptake

* Enzymatic Activities

WP5 - MICROBIAL METABOLIC ACTIVITIESCNRS DR 12 - LMM


Microbial activities

* BRINES

* INTERFACE (Seawater-Brine)

Seawater, Upper & Lower Interface (2001)

Along Interface (2002)

* BRINES

Brines, Upper Brine, Heavy Brine (Urania)

* MUD PIT (Urania)

* SEDIMENT

* OVERLAYING BRINE (or BOTTOM BRINE)

* SEDIMENT (Levels A, B, C)

MICROBIAL ACTIVITIES

BIODEEP CRUISES 2001 & 2002

L’ATALANTE, BANNOCK, DISCOVERY, URANIA


* METHANE PRODUCTION (MP)

CH4 produced (µmol/L)

* NATURAL MP & POTENTIAL MP

* No Amendment & Pool of Substrates

* BIODEEP-3 CRUISE (2002)

* MPR = Methane Production Rate

* MP ± Sulfate Reduction inhibitor (Molybdate)

* SULFATE REDUCTION SRR

35SH2 produced from 35SO4 (µmol/L/d)

* BIODEEP-3 CRUISE 2002

* SR ± Methane Production inhibitor (BES)

STRICT ANAEROBIC ACTIVITIES


Bacterial production activities

* BACTERIAL PRODUCTION(Biomass Production)

* Incorporation of 3H-Leucine

* ENZYMATIC ACTIVITIES BIODEEP-2 Cruise 2001

* Aminopeptidase

* Phosphatase

* GLUTAMATE UPTAKE

* Assimilation & Respiration of 14C-Glutamate

* GLUCOSE UPTAKE BIODEEP-3 Cruise 2002

* Assimilation & Respiration of 14C-Glucose

BACTERIAL PRODUCTION & ACTIVITIES


L atalante

Discovered in 1993

Hypersaline S‰ ~10-times SW

Brine layer homogeneous

Stratification T°C - 3 layers

L’ATALANTE

DISCOVERY

URANIA

psu nM mM

T°C S‰ CH4 SO4

13.84 39 2-6 31

13.82

13.91 400. 103

14.06 320 188. 103 333

SEAWATER

3295 m

INTERFACE

SW

UPPER INTERFACE01

LOWER INTERFACE 01

40 m 30 m 16 m

BRINE

BODY BRINE01 & 02

NaCl dominant (180 g Cl/L)

Potassium up to 390 mM

BOTTOM BRINE

BOTTOM BRINE01 & 02

SEDIMENT LEVEL A01 & 02

SEDIMENT LEVEL B 01 & 02

SEDIMENT

3390 m

L’ATALANTE


Salinity S‰ (PSU)

Dissolved [CH4] (µM)

S‰ & [CH4]

MP nat & MP pot

SRR & MPR

INTERFACE

BRINE-01

BRINE-02

BOTTOM BRINE

SED-A

SED-B

INTERFACE

BRINE-01

BRINE-02

BOTTOM BRINE

SED-A

SED-B

INTERFACE

BRINE-01

BRINE-02

BOTTOM BRINE

SED-A

SED-B

Sulfate Reduction Rate (µM H2S/d)

0 200 400 600 0 50 100 150 0 50 100 150

Methane Production Natural (µM CH4)

Methane Production Potential (µM CH4)

Methane Production Rate (µM CH4/d)

L’ATALANTE BASINS‰, [CH4], MP & SR


Bannock basin libeccio

2 layers: Upper Brine

Lower Brine

psu nM mM mM

pH S‰ T°C CH4 H2S SO4

8.2 39 13.5 2-6 0 31

6.5 321 14.41 128.103 1.34 99

326 15.14 2.7 130

Bottom Brine 37.103

SEAWATER

SW

3276 db

INTERFACE 1

UPPER INTERFACE01

LOWER INTERFACE01

INTERFACE 02

140 dbar

UPPER BRINE

BODY BRINE01 & 02

INTERFACE 2

LOWER BRINE

300 dbar

BOTTOM BRINE

BOTTOM BRINE01 & 02

SEDIMENT-A01 & 02

SEDIMENT-B 02

SEDIMENT- C 02

3778 db

SEDIMENT

BANNOCK BASIN LIBECCIO


Salinity S‰ (PSU)

Dissolved [CH4] (µM)

S‰ & [CH4]

BRINE-01

BRINE-02

BOTTOM BRINE

SED-A

SED-B

MP nat & MP pot

SRR & MPR

BRINE-01

BRINE-02

BOTTOM BRINE

SED-A

SED-B

100

INTERFACE

INTERFACE

80

60

3 340

40

Depth (db)

588

20

3 345

0

0

50

100

150

Sulfate Reduction Rate (µM H2S/d)

100

200

300

400

0

Methane Production Natural (µM CH4)

Methane Production Potential (µM CH4)

Methane Production Rate (µM CH4/d)

BANNOCK BASIN LIBECCIO sub-basinS‰, MP, [CH4] & SR


Salinity S‰ (PSU)

Salinity S‰ (PSU)

Dissolved [CH4] (µM)

Dissolved [CH4] (µM)

S‰ & [CH4]

S‰ & [CH4]

MP nat & MP pot

50

40

300

30

INTERFACE SW / BRINE

3 340

Depth (db)

20

200

3 341

3 341

3 345

3 339

3 339

3 340

3 340

3 342

3 342

3 343

3 343

10

0

100

0

100

200

300

400

0

Methane Production Natural (µM CH4)

Methane Production Potential (µM CH4)

BANNOCK BASIN INTERFACE SW / BRINE

S‰, [CH4] & MP


Discovery

Discovered in 1993-94 - Hypersaline

Area = 7.5 km2 - Volume = 2 x 108 m3

Brine layer homogeneous

L’ATALANTE

DISCOVERY

URANIA

psu nM mM mol/Kg

S‰ pH CH4 SO4 Na Mg Cl

39 7.8 2-6 31 0.486 0.06 0.57

>120 4.5 36.103 85 0.008 5.15 10.15

SEAWATER

3 516 m

INTERFACE

SW

UPPER INTERFACE01

LOWER INTERFACE01

INTERFACE 02

BRINE

BODY BRINE01 & 02

BOTTOM BRINE

SEDIMENT- A01 & 02

SEDIMENT-B 02

SEDIMENT-C 02

SEDIMENT

3 533 m

DISCOVERY


Salinity S‰/10

SEAWATER

BRINE-01

BRINE-02

SED-A

SED-B

SED-C

Dissolved [CH4] (µM)

S‰ & [CH4]

MP nat & MP pot

SRR & MPR

SEAWATER

BRINE-01

BRINE-02

SED-A

SED-B

SED-C

INTERFACE

INTERFACE

3 650

Depth (db)

3 655

3 660

320

Sulfate Reduction Rate (µM H2S/d)

0 10 20 30 40 50 60 70 0 10 20 30 40 50 60 0 10 20 30 40 50 60

Methane Production Natural (µM CH4)

Methane Production Potential (µM CH4)

Methane Production Rate (µM CH4/d)

DISCOVERY BASINS‰, [CH4], MP & SR


Salinity S‰/10

Salinity S‰/10

S‰ & [CH4]

Dissolved [CH4] (µM)

Dissolved [CH4] (µM)

10

3 652

3 652

3 653

3 653

3 654

3 654

3 655

3 655

3 656

3 656

3 657

3 657

S‰ & [CH4]

MP nat & MP pot

8

6

70

4

60

3 650

50

2

40

3 655

INTERFACE SW / BRINE

0

Depth (db)

30

3 660

20

10

0

0 10 20 30 40 50 60 70

Methane Production Natural (µM CH4)

Methane Production Potential (µM CH4)

DISCOVERY INTERFACE SW / BRINE S‰, [CH4] & MP


Urania

WEST

MIDDLE

Discovered in 1993 - 6 km long

Hypersaline S‰ ~5-8-times SW

Horseshoe-shaped depression

3 layers: Brine, Heavy Brine, Mud pit

EAST

psu nM mM mM

pH S‰ T°C CH4 H2S SO4

8.6 39 13.74 2-6 0 31

6.8 200 15.78 2692.103 10 85

210

45.3

SEAWATER

SW

UPPER INTERFACE 01

LOWER INTERFACE01

INTERFACE 02

3462 m

INTERFACE 1

EAST-BRINE 01

WEST-BRINE 02

BRINE

80 - 200 m

INTERFACE 2

EAST-INTERFACE-2 02

EAST-HEAVY BRINE 02

HEAVY BRINE

WEST-MUD PIT 02

MUD PIT

3714 m

MUD PIT

BOTTOM BRINE01

SEDIMENT-A01 & 02

SEDIMENT-B01 & 02

BOTTOM BRINE

3850 m

SEDIMENT

URANIA


[CH4]/10

Salinity S‰ (PSU)

Dissolved [CH4]/10 (µM)

S‰ & [CH4]

MP nat & MP pot

SRR & MPR

3 520

INTERFACE-1

BRINE-01

BRINE-02

INTERFACE-2

HEAVY BRINE

MUD PIT

BOTTOM BRINE

SED-A

SED-B

INTERFACE-1

BRINE-01

BRINE-02

INTERFACE-2

HEAVY BRINE

MUD PIT

BOTTOM BRINE

SED-A

SED-B

3 540

Depth (db)

3 600

3 620

1219 1797

Sulfate Reduction Rate (µM H2S/d)

0 250 500 750 1000 0 250 500 750 1000 0 50 100 150 200 250 300

Methane Production Natural (µmol CH4/l)

Methane Production Potential (µmol CH4/l)

Methane Production Rate (µmol CH4/l/d

URANIA BASINS‰, [CH4], MP & SR


[CH4]/10

Salinity S‰ (PSU)

Salinity S‰ (PSU)

3 608

3 608

3 606

3 606

3 610

3 610

3 612

3 612

3 614

3 614

3 616

3 616

Dissolved [CH4]/10 (µM)

Dissolved [CH4]/10 (µM)

2000

3 520

3 520

3 524

3 524

3 528

3 528

3 532

3 532

3 536

3 536

3 540

3 540

MP nat & MP pot

SRR & MPR

S‰ & [CH4]

1500

3 520

1000

750

INTERFACE

SW / BRINE

3 540

500

1000

500

0

750

250

Depth (db)

500

3 600

0

500

250

400

INTERFACE-2

BRINE / HEAVY BRINE

3 620

0

300

200

100

0 250 500 750 1000

Methane Production Natural (µmol CH4/l)

0

Methane Production Potential (µmol CH4/l)

URANIA BASIN INTERFACES S‰, [CH4] & MP


L atalante bannock discovery urania s ch4

700

6000

600

ATALANTE

5000

BANNOCK

500

DISCOVERY

4000

URANIA

S‰

[CH4]

400

[CH4]

3000

300

BRINE

BOTTOM

BRINE

2000

200

100

1000

INTERFACE

0

0

[CH4] µM

S‰ (psu)

0

100

200

300

400

500

600

[CH4] µM

L'ATALANTE, BANNOCK, DISCOVERY & URANIAS‰ & [CH4]


L atalante bannock discovery urania methane production

1000

MP = METHANE PRODUCTION

µmol CH4 produced per liter

ATALANTE

BANNOCK

800

DISCOVERY

URANIA

600

BRINE

BOTTOM

BRINE

SED-A

SED-B

SED-C

200

MPR = METHANE PRODUCTION RATE

µmol CH4 produced per liter per day

INTERFACE

400

150

200

BRINE

BOTTOM

BRINE

SED-A

SED-B

SED-C

100

0

50

0

MP Natural : [CH4] µM

MPR Natural : [CH4] µM/d

L'ATALANTE, BANNOCK, DISCOVERY & URANIAMETHANE PRODUCTION


L atalante bannock discovery urania methane production1

60

MP = METHANE PRODUCTION

µmol CH4 produced per liter

ATALANTE

BANNOCK

50

DISCOVERY

35

40

BRINE

BOTTOM

BRINE

SED-A

SED-B

SED-C

INTERFACE

30

30

MPR = METHANE PRODUCTION RATE

µmol CH4 produced per liter per day

25

20

20

10

BRINE

BOTTOM

BRINE

SED-A

SED-B

SED-C

15

0

10

5

0

MPR Natural : [CH4] µM/d

MP Natural : [CH4] µM

L'ATALANTE, BANNOCK, DISCOVERY & URANIAMETHANE PRODUCTION


Urania basin methane production inhibitor of sr

MPR = METHANE PRODUCTION RATE

µmol CH4 produced per liter per day

MPR in the presence or absence of Molybdate

(inhibitor of SR)

300

without MOB

with MOB

250

200

150

MPnat: CH4 µM/l

100

50

0

BrineBrineMudPitMudpit Sed-BSed-B

+MOB +MOB +MOB

x 17 x 1.5 x 5.7

URANIA BASINMETHANE PRODUCTION ± INHIBITOR OF SR


L atalante bannock discovery urania sulfate reduction

350

300

ATALANTE

250

SRR = SULFATE REDUCTION RATE

µmol H2S produced per liter per day

BANNOCK

200

DISCOVERY

150

URANIA

150

125

100

BRINE

BOTTOM

BRINE

SED-A

SED-B

SED-C

100

50

INTERFACE

0

75

50

25

0

SRR : H2S mM/d

SRR : H2S mM/d

L'ATALANTE, BANNOCK, DISCOVERY & URANIASULFATE REDUCTION


L atalante bannock discovery urania enzymatic activities

ATALANTE

BANNOCK

DISCOVERY

URANIA

OXIC

SW 3 000 m

PHOSPHATASE ACTIVITY

nmol produced per liter per hour

AMINO-PEPTIDASE ACTIVITY

nmol produced per liter per hour

INTERFACE

BRINE

BOTTOM BRINE

SEDIMENT-A

SEDIMENT-B

SW 3 000 m

INTERFACE

BRINE

BOTTOM BRINE

SEDIMENT-A

SEDIMENT-B

0.1

0.1

1

1

10

10

100

100

1000

1000

10000

10000

100000

100000

pmol/l/h

pmol/l/h

L'ATALANTE, BANNOCK, DISCOVERY & URANIAENZYMATIC ACTIVITIES


L atalante bannock discovery urania glutamate glucose uptake

ATALANTE

100000

BANNOCK

DISCOVERY

10000

URANIA

SW

1000

GLUCOSE UPTAKE

pmol per liter per hour

GLUTAMATE UPTAKE

pmol per liter per hour

100

10

Ligurian Sea 2000 m

SW

INTERFACE

BRINE

BOTTOM BRINE

SEDIMENT-A

SEDIMENT-B

SW

BRINE

BOTTOM BRINE

SEDIMENT-A

SEDIMENT-B

1

0.1

1

10

100

1000

10000

100000

1000000

pmol/l/h

pmol/l/h

L'ATALANTE, BANNOCK, DISCOVERY & URANIAGLUTAMATE & GLUCOSE UPTAKE


L atalante bannock discovery urania bacterial production

Brine-DC

Brine-HP

BACTERIAL PRODUCTION

ng C per liter per day

BACTERIAL PRODUCTION 2001

ng C per liter per day

SW 3000 m

Interface

Brine

DC = Samples Decompressed

HP = Samples Ambient Pressure

0.8

SW

ATALANTE

BANNOCK

DISCOVERY

URANIA

0.6

2

0.4

1.5

0.2

1

0

0.5

SW

ATALANTE

BANNOCK

DISCOVERY

URANIA

0

L'ATALANTE, BANNOCK, DISCOVERY & URANIABACTERIAL PRODUCTION


L atalante bannock discovery urania effect of pressure conditions on bacterial activities

AA = Aminopeptidase Activity

PA = Phophatase Activity

GLUT = Glutamate Uptake

BACTERIAL ACTIVITIES

DC = Samples Decompressed

HP = Samples Ambient Pressure

SEAWATER ATALANTE BANNOCK

30

DC

56.30

HP

20

10

0

AA GLUT AA AA PA GLUT

SEAWATER ATALANTE BANNOCK

L'ATALANTE, BANNOCK, DISCOVERY & URANIAEFFECT OF PRESSURE CONDITIONS ONBACTERIAL ACTIVITIES


Conclusions

* PRESENCE OF ACTIVEARCHAEBACTERIA & BACTERIA

IN THE DHABs

* INTERFACE

* BRINES

* SEDIMENT

* BACTERIA ADAPTED TO IN SITU CONDITIONS

(HIGH PRESSURE, HIGH SALINITY, ANOXIA...)

AS ACTIVITY IS HIGHER UNDER AMBIENT

CONDITIONS THAN IN DECOMPRESSED SAMPLES

CONCLUSIONS


Conclusions1
CONCLUSIONS

* GLOBALLY, IN THE DIFFERENT DHABs

SRL'ATALANTE>DISCOVERY >URANIA≥ BANNOCK

MPURANIA>BANNOCK≥ L'ATALANTE>DISCOVERY

BABANNOCK>URANIA>L'ATALANTE>DISCOVERY

AABANNOCK>L'ATALANTE>URANIA>DISCOVERY

PABANNOCK>L'ATALANTE>URANIA>DISCOVERY

GLUTBANNOCK>L'ATALANTE>URANIA>DISCOVERY

GLUCURANIA>DISCOVERY>L'ATALANTE>BANNOCK


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