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WP5 - MICROBIAL METABOLIC ACTIVITIES CNRS DR 12 - LMM BACTERIAL ACTIVITIES * STRICT ANAEROBIC ACTIVITIES * Methane Production (MP) * Sulfate Reduction.

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Presentation on theme: "WP5 - MICROBIAL METABOLIC ACTIVITIES CNRS DR 12 - LMM BACTERIAL ACTIVITIES * STRICT ANAEROBIC ACTIVITIES * Methane Production (MP) * Sulfate Reduction."— Presentation transcript:

1 WP5 - MICROBIAL METABOLIC ACTIVITIES CNRS DR 12 - LMM BACTERIAL ACTIVITIES * STRICT ANAEROBIC ACTIVITIES * Methane Production (MP) * Sulfate Reduction (SR) * BACTERIAL PRODUCTION & ACTIVITY * Bacterial Production (BP) * Glutamate & Glucose Uptake * Enzymatic Activities D. MARTY, J. GARCIN, C. TAMBURINI, A. BIANCHI BIODEEP Meeting VIII VENEZIA 13-15 October 2003

2 MICROBIAL ACTIVITIES BIODEEP CRUISES 2001 & 2002 L’ATALANTE, BANNOCK, DISCOVERY, URANIA * 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)

3 STRICT ANAEROBIC ACTIVITIES * METHANE PRODUCTION (MP) CH 4 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 REDUCTIONSRR 35 SH 2 produced from 35 SO 4 (µmol/L/d) * BIODEEP-3 CRUISE 2002 * SR ± Methane Production inhibitor (BES)

4 BACTERIAL PRODUCTION & ACTIVITIES * BACTERIAL PRODUCTION(Biomass Production) * Incorporation of 3 H-Leucine * ENZYMATIC ACTIVITIES BIODEEP-2 Cruise 2001 * Aminopeptidase * Phosphatase * GLUTAMATE UPTAKE * Assimilation & Respiration of 14 C-Glutamate * GLUCOSE UPTAKE BIODEEP-3 Cruise 2002 * Assimilation & Respiration of 14 C-Glucose

5 L’ATALANTE URANIA L’ATALANTE DISCOVERY Discovered in 1993 HypersalineS‰ ~10-times SW Brine layer homogeneous Stratification T°C - 3 layers UPPER INTERFACE 01 LOWER INTERFACE 01 BODY BRINE 01 & 02 BOTTOM BRINE 01 & 02 SEDIMENT LEVEL A 01 & 02 SEDIMENT LEVEL B 01 & 02 SEAWATER INTERFACE BRINE BOTTOM BRINE SEDIMENT 3390 m 3295 m psu nM mM T°CS‰ CH 4 SO 4 13.84 39 2-6 31 13.82 13.91 400. 10 3 14.06 320 188. 10 3 333 40 m 30 m 16 m NaCl dominant (180 g Cl/L) Potassium up to 390 mM SW

6 L’ATALANTE BASIN S‰, [CH4], MP & SR Salinity S‰ (PSU) Dissolved [CH4] (µM) Methane Production Natural (µM CH4) Methane Production Potential (µM CH4) Methane Production Rate (µM CH4/d) Sulfate Reduction Rate (µM H2S/d) 0 200 400 600 0 50 100 150 0 50 100 150 INTERFACE BRINE-01 BRINE-02 BOTTOM BRINE SED-A SED-B INTERFACE BRINE-01 BRINE-02 BOTTOM BRINE SED-A SED-B S‰ & [CH4]MP nat & MP potSRR & MPR INTERFACE BRINE-01 BRINE-02 BOTTOM BRINE SED-A SED-B

7 BANNOCK BASINLIBECCIO 2 layers:Upper Brine Lower Brine UPPER INTERFACE 01 LOWER INTERFACE 01 INTERFACE 02 BODY BRINE 01 & 02 BOTTOM BRINE 01 & 02 SEDIMENT-A 01 & 02 SEDIMENT-B 02 SEDIMENT- C 02 3778 db 3276 db 300 dbar 140 dbar SEAWATER INTERFACE 1 UPPER BRINE BOTTOM BRINE SEDIMENT INTERFACE 2 LOWER BRINE psu nM mM mM pH S‰ T°C CH 4 H 2 S SO 4 8.2 39 13.5 2-6 031 6.5 321 14.41 128.10 3 1.3499 326 15.14 2.7 130 Bottom Brine 37.10 3 SW

8 BANNOCK BASIN LIBECCIO sub-basin S‰, MP, [CH4] & SR Salinity S‰ (PSU) Dissolved [CH4] (µM) Methane Production Natural (µM CH4) Methane Production Potential (µM CH4) Methane Production Rate (µM CH4/d) Sulfate Reduction Rate (µM H2S/d) Depth (db) 3 340 3 345 588 0 100200300400050100150 0 2040 60 80 100 INTERFACE BRINE-01 BRINE-02 BOTTOM BRINE SED-A SED-B INTERFACE BRINE-01 BRINE-02 BOTTOM BRINE SED-A SED-B S‰ & [CH4]MP nat & MP potSRR & MPR

9 Salinity S‰ (PSU) Dissolved [CH4] (µM) Depth (db) 3 340 3 345 0 100200300400 Methane Production Natural (µM CH4) Methane Production Potential (µM CH4) 0 100200300 Salinity S‰ (PSU) Dissolved [CH4] (µM) 0 1020304050 3 339 3 340 3 341 3 342 3 343 3 339 3 340 3 341 3 342 3 343 INTERFACE SW / BRINE S‰ & [CH4] MP nat & MP pot BANNOCK BASIN INTERFACE SW / BRINE S‰, [CH4] & MP

10 DISCOVERY Discovered in 1993-94 - Hypersaline Area = 7.5 km 2 - Volume = 2 x 10 8 m 3 Brine layer homogeneous URANIA L’ATALANTE DISCOVERY SW psu nM mM mol/Kg S‰ pH CH 4 SO 4 Na Mg Cl 39 7.8 2-6 31 0.486 0.06 0.57 >120 4.5 36.10 3 85 0.008 5.15 10.15 SEAWATER INTERFACE BRINE BOTTOM BRINE SEDIMENT 3 516 m 3 533 m UPPER INTERFACE 01 LOWER INTERFACE 01 INTERFACE 02 BODY BRINE 01 & 02 SEDIMENT- A 01 & 02 SEDIMENT-B 02 SEDIMENT-C 02

11 DISCOVERY BASIN S‰, [CH4], MP & SR 0 10 20 30 40 50 60 70 0 10 20 30 40 50 60 0 10 20 30 40 50 60 3 650 3 655 3 660 Depth (db) Salinity S‰/10 Dissolved [CH4] (µM) Methane Production Natural (µM CH4) Methane Production Potential (µM CH4) Methane Production Rate (µM CH4/d) Sulfate Reduction Rate (µM H2S/d) 320 INTERFACE SEAWATER BRINE-01 BRINE-02 SED-A SED-B SED-C INTERFACE SEAWATER BRINE-01 BRINE-02 SED-A SED-B SED-C S‰ & [CH4]MP nat & MP potSRR & MPR

12 DISCOVERY INTERFACE SW / BRINE S‰, [CH4] & MP 0 10 20 30 40 50 60 70 3 650 3 655 3 660 Depth (db) Salinity S‰/10 Dissolved [CH4] (µM) Methane Production Natural (µM CH4) Methane Production Potential (µM CH4) Salinity S‰/10 Dissolved [CH4] (µM) 3 652 3 653 3 654 3 655 3 656 3 657 0 1020304050607002468 10 3 652 3 653 3 654 3 655 3 656 3 657 INTERFACE SW / BRINE S‰ & [CH4] MP nat & MP pot

13 URANIA Discovered in 1993 - 6 km long Hypersaline S‰ ~5-8-times SW Horseshoe-shaped depression 3 layers: Brine, Heavy Brine, Mud pit WEST EAST MIDDLE 3714 m 80 - 200 m 3462 m 3850 m SEAWATER INTERFACE 1 BRINE BOTTOM BRINE SEDIMENT HEAVY BRINE INTERFACE 2 MUD PIT UPPER INTERFACE 01 LOWER INTERFACE 01 INTERFACE 02 EAST-BRINE 01 WEST-BRINE 02 EAST-INTERFACE-2 02 EAST-HEAVY BRINE 02 WEST-MUD PIT 02 BOTTOM BRINE 01 SEDIMENT-A 01 & 02 SEDIMENT-B 01 & 02 psu nM mM mM pH S‰ T°C CH 4 H 2 S SO 4 8.6 39 13.74 2-6 0 31 6.8 200 15.78 2692.10 3 10 85 210 45.3 SW MUD PIT

14 URANIA BASIN S‰, [CH4], MP & SR Depth (db) 0 250 500 750 1000 0 250 500 750 1000 0 50 100 150 200 250 300 [CH4]/10 3 520 3 540 3 600 3 620 Salinity S‰ (PSU) Dissolved [CH4]/10 (µM) Methane Production Natural (µmol CH4/l) Methane Production Potential (µmol CH4/l) Methane Production Rate (µmol CH4/l/d Sulfate Reduction Rate (µM H2S/d) 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 1219 1797 S‰ & [CH4]MP nat & MP potSRR & MPR

15 URANIA BASIN INTERFACES S‰, [CH4] & MP 0 250 500 750 1000 Depth (db) [CH4]/10 3 520 3 540 3 600 3 620 Salinity S‰ (PSU) Dissolved [CH4]/10 (µM) INTERFACE SW / BRINE INTERFACE-2 BRINE / HEAVY BRINE S‰ & [CH4] Methane Production Natural (µmol CH4/l) Methane Production Potential (µmol CH4/l) 0 2505007501000 3 606 3 608 3 610 3 612 3 614 3 616 Salinity S‰ (PSU) Dissolved [CH4]/10 (µM) 0 500100015002000 3 520 3 524 3 528 3 532 3 536 3 540 0 100200300400500 3 520 3 524 3 528 3 532 3 536 3 540 0 250500750 3 606 3 608 3 610 3 612 3 614 3 616 MP nat & MP potSRR & MPR

16 L'ATALANTE, BANNOCK, DISCOVERY & URANIA S‰ & [CH4] ATALANTE BANNOCK DISCOVERY URANIA S‰ 0 100200300400500600700 S‰ (psu) [CH4] 0 100020003000400050006000 [CH4] µM 0100200300400500 600 [CH4] µM [CH4] BRINE BOTTOM BRINE INTERFACE

17 L'ATALANTE, BANNOCK, DISCOVERY & URANIA METHANE PRODUCTION ATALANTE BANNOCK DISCOVERY URANIA MP = METHANE PRODUCTION µmol CH4 produced per liter MP Natural : [CH4] µM 0 2004006008001000 MPR Natural : [CH4] µM/d MPR = METHANE PRODUCTION RATE µmol CH4 produced per liter per day 0 50100150200 BRINE BOTTOM BRINE SED-A SED-B SED-C BRINE BOTTOM BRINE SED-A SED-B SED-C INTERFACE

18 L'ATALANTE, BANNOCK, DISCOVERY & URANIA METHANE PRODUCTION ATALANTE BANNOCK DISCOVERY 0 102030405060 MP Natural : [CH4] µMMPR Natural : [CH4] µM/d 05 101520253035 MP = METHANE PRODUCTION µmol CH4 produced per liter MPR = METHANE PRODUCTION RATE µmol CH4 produced per liter per day BRINE BOTTOM BRINE SED-A SED-B SED-C BRINE BOTTOM BRINE SED-A SED-B SED-C INTERFACE

19 URANIA BASIN METHANE PRODUCTION ± INHIBITOR OF SR MPR in the presence or absence of Molybdate (inhibitor of SR) MPR = METHANE PRODUCTION RATE µmol CH4 produced per liter per day Brine Brine MudPit Mudpit Sed-B Sed-B +MOB +MOB +MOB x 17 x 1.5 x 5.7 MPnat: CH4 µM/l 0 50 100 150 200 250 300 without MOB with MOB

20 L'ATALANTE, BANNOCK, DISCOVERY & URANIA SULFATE REDUCTION 0 255075100125150 SRR : H2S mM/d ATALANTE BANNOCK DISCOVERY URANIA SRR : H2S mM/d 0 50100150200250300350 SRR = SULFATE REDUCTION RATE µmol H2S produced per liter per day BRINE BOTTOM BRINE SED-A SED-B SED-C INTERFACE

21 BRINE BOTTOM BRINE SEDIMENT-A SEDIMENT-B AMINO-PEPTIDASE ACTIVITY nmol produced per liter per hour pmol/l/h 0.1110100100010000100000 SW 3 000 m ATALANTE BANNOCK DISCOVERY URANIA OXIC PHOSPHATASE ACTIVITY nmol produced per liter per hour INTERFACE BRINE BOTTOM BRINE SEDIMENT-A SEDIMENT-B pmol/l/h 0.1110100100010000100000 SW 3 000 m L'ATALANTE, BANNOCK, DISCOVERY & URANIA ENZYMATIC ACTIVITIES

22 GLUCOSE UPTAKE pmol per liter per hour SW BRINE BOTTOM BRINE SEDIMENT-A SEDIMENT-B pmol/l/h 1 101001000 10000 100000 Ligurian Sea 2000 m ATALANTE BANNOCK DISCOVERY URANIA SW GLUTAMATE UPTAKE pmol per liter per hour SW INTERFACE BRINE BOTTOM BRINE SEDIMENT-A SEDIMENT-B pmol/l/h 0.11101001000100001000001000000 L'ATALANTE, BANNOCK, DISCOVERY & URANIA GLUTAMATE & GLUCOSE UPTAKE

23 L'ATALANTE, BANNOCK, DISCOVERY & URANIA BACTERIAL PRODUCTION BACTERIAL PRODUCTION ng C per liter per day SWATALANTEBANNOCKDISCOVERYURANIA 0 0.2 0.4 0.6 0.8 Interface Brine SW 3000 m BACTERIAL PRODUCTION 2001 ng C per liter per day DC = Samples Decompressed HP = Samples Ambient Pressure SWATALANTEBANNOCKDISCOVERYURANIA 0 0.5 1 1.5 2 Brine-DC Brine-HP

24 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 DC HP 0 10 20 30 56.30 SEAWATER ATALANTE BANNOCK AAGLUTAA AA PA GLUT SEAWATER ATALANTE BANNOCK

25 CONCLUSIONS * PRESENCE OF ACTIVE ARCHAEBACTERIA & 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

26 CONCLUSIONS * GLOBALLY, IN THE DIFFERENT DHABs SR L'ATALANTE > DISCOVERY > URANIA ≥ BANNOCK MP URANIA > BANNOCK ≥ L'ATALANTE > DISCOVERY BA BANNOCK > URANIA > L'ATALANTE > DISCOVERY AABANNOCK > L'ATALANTE > URANIA > DISCOVERY PABANNOCK > L'ATALANTE > URANIA > DISCOVERY GLUT BANNOCK > L'ATALANTE > URANIA > DISCOVERY GLUC URANIA > DISCOVERY > L'ATALANTE > BANNOCK

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