wp5 - microbial metabolic activities cnrs dr 12 - lmgem

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WP5 - MICROBIAL METABOLIC ACTIVITIESCNRS DR 12 - LMGEM

BACTERIAL ACTIVITIES

* STRICT ANAEROBIC ACTIVITIES* Methane Production (MP)* Sulfate Reduction (SR)

* BACTERIAL PRODUCTION & ACTIVITY* Bacterial Production (BP)* Glutamate Uptake* Chemotrophic production* Hexadecane respiration

Contract EVK3-2000-00042

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

BIODEEP Meeting IX MILANO 17-19 May 2004

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Microbial activities measured Glutamate uptake, Bacterial & Chemolithotrophic production,

Hexadecane respiration

* Heterotrophic bacterial production representative of the processes due to the whole heterotrophic microbial community

* Chemolithotrophic production biomass production by chemolitrophic organisms growing without organic enrichment, and using CO2 as C source for biomass production * Assimilation and respiration of glutamate used as a rough index of the ratio linking bacterial biomass production processes to those used to produce energy * Hexadecane respiration occasionally measured as a paragon of ability for microbial populations to mineralize hydrocarbon compounds

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Bacterial production in the Upper interfacesrelatively to the salinity

0

5

10

15

20

25

30

35

40

0 10 20 30 40 50

pmol C l-1 h-1

S (

%)

Urania

L'Atalante

Bannock

Discovery

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Microbial activities within brines Heterotrophic bacterial & Chemolithotrophic production

Heterotrophic bacterial production

0

2

4

6

8

10

12

14

16

Urania L’Atalante Discovery Bannock

pm

ol C

l-1

h-1

Chemolitotrophic production

ND ND

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Microbial activities within brines Glutamate assimilation and respiration, Hexadecane

respiration

00,20,40,60,8

11,21,41,61,8

2

pm

ol

C l

-1 h

-1

00,20,40,60,8

11,21,41,61,8

2

Urania L’Atalante Discovery Bannock Urania L’Atalante Discovery Bannock

ND ND

Glutamate assimilationGlutamate respiration


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Microbial activities in near-bottom water

Heterotrophic bacterial & Chemolithotrophic production

Heterotrophic bacterial production

Chemolitotrophic production

0

5

10

15

20

25

30

35

pm

ol C

l-1

h-1


ND NDNo NBB available

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Microbial activities in near-bottom water

Glutamate assimilation and respiration, Hexadecane respiration

0

0,5

1

1,5

2

2,5

3

3,5

4

pm

ol

C l

-1 h

-1

0

0,5

1

1,5

2

2,5

3

3,5

4

Urania L’Atalante Discovery Bannock Urania L’Atalante Discovery Bannock

ND No NBB available No NBB availableND

Glutamate assimilationGlutamate respiration


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Microbial activities in the superficial sediments

Glutamate assimilation and respiration, Hexadecane respiration

Glutamate respiration with 1 µM initial concentration

Glutamate respiration with 5 µM initial concentration

0

1000

2000

3000

4000

5000

6000

pm

ol C

l-1

h-1

Hexadecane respiration with 5 µM initial concentration


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Toward the research of piezophile hydrocarbonoclastic bacteria from the

Urania Basin

* HP 1 10 mL inoculum brine + 100 mL sterilized brine + 40 µL Hexadecane + 20 mg (NH4)3PO4 + 8mM Na2S (ANOXIC CONDITIONS)

* HP 2 10 mL inoculum interface + 100 mL sterilized interface + 40 µL Hexadecane + 20 mg (NH4)3PO4

* HP 3 10 mL inoculum brine + 100 mL ONR7a + 40 µL Hexadecane + 20 mg (NH4)3PO4

* HP 4 10 mL inoculum interface + 100 mL ONR7a + 40 µL Hexadecane + 20 mg (NH4)3PO4

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Urania Basin

Inoculum(10 mL)

Medium(100 mL)

Oxic conditio

n

Hexadecane

(NH4)3PO4

HP 1 BrineStrilized

brineANOXIC(8mM Na2S)

40 µL 20 mg

HP 2 InterfaceStrilized interface

OXIC 40 µL 20 mg

HP 3 Brine ONR7a OXIC 40 µL 20 mg

HP4 Interface ONR7a OXIC 40 µL 20 mg

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Urania Basin

Pressure = 35 MPa

Agitation table

HP bottles

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Urania Basin

Results from Giuseppe D’Auria

After several months of incubation RNA (16S cDNA) extraction of subsamples of main culture show that an active population is living and growing in high-pressure conditions in each condition tested.

100bp Marker

HP bottles1 2 3 4

Cloning and sequencing are in progress…

PCR gel of 16s cDNA amplification

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What about our article in AEM?

Submission in September 2002

Response in January 2003 Major revisions : improve the English language, physical and chemical parameters, numbers of bacteriaPotential authors : Gasparonni, D’Auria for TC, chemist?,

May 2004 It would be very important to include other authors to improve the paper and corroborate some hypotheses we did


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Data from cruise I

Focus: Evidence of microbial life in DHABs


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wp5 - microbial metabolic activities cnrs dr 12 - lmgem

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