WP3WP3WP3WP3

State of the Art May 2004State of the Art May 2004

CNRCNRIAMC - Istituto per l’Ambiente Marino IAMC - Istituto per l’Ambiente Marino

CostieroCostieroMessina, ItalyMessina, Italy

Università degli Studi di MessinaUniversità degli Studi di Messina

CNRCNRIAMC - Istituto per l’Ambiente Marino IAMC - Istituto per l’Ambiente Marino

CostieroCostieroMessina, ItalyMessina, Italy

Università degli Studi di MessinaUniversità degli Studi di Messina

16 rRNA Distribution at Family level

In study: >830 sequences analysed

9 levels for Urania from 3.8% of Salinity to BB

7 Levels for Bannock from 3.9% to 28% of salinity

8 Levels for Discovery from 6.2% of salinity to BB

7 Levels for L’Atalante from 4% of salinity to BB

1st Statistical aproach:1st Statistical aproach: Family level (-aceae)Family level (-aceae)

2nd Statistical aproach: Division Level

9,8

12,8

16,91920,6

23,2

28

24.622.4

1110,2

15,414,4

64,2 3,9

0

5

10

15

20

25

30

-300 -250 -200 -150 -100 -50 0 501,3351,3401,3451,3501,3551,3601,3651,3701,3751,3801,3851,390

16S rRNA Bannock Sampling ProfileS

alin

ity %

44,84,45,27,8

12,8

19,421,2

23,824,225,625,426

27,6

0

5

10

15

20

25

30

-350 -300 -250 -200 -150 -100 -50 0Eh

1,335

1,34

1,345

1,35

1,355

1,36

1,365

1,37

1,375

1,38

1,385

6,2779

1315,4

18

21,2

2424,4

0

5

10

15

20

25

30

-300 -250 -200 -150 -100 -50 0 50Eh

1,34

1,35

1,36

1,37

1,38

1,39

1,4

1,41

3,8

6,77,78,28,79

10,910,9

13,413,91515,3

5,1

15,614,616

0

5

10

15

20

-350 -250 -150 -50 50Eh

1,338

1,34

1,342

1,344

1,346

1,348

1,35

1,352

1,354

1,356

1,358

1,36

Re

frac

tion

Ind

ex

Eh

L’AtalanteL’AtalanteDiscoveryDiscovery

UraniaUrania BannockBannock

DHABs Diversity (families)

Urania

Bannock

Discovery

L'Atalante

Stress: 0,18

nMDS analysisnMDS analysis

Urania

Bannock

Discovery

L'Atalante

U3.8

U5.1

U8.2

U8.7

U10.9

U14U14.6

U15.6

UBB

B3.9B12.8

B14.4

B16.9

B22.4B24.6

B28

D6.2

D9

D15.4D17*

D21.2

D24.4

D26*

DBB

A4

A7,8A12,8

A19,4A23,8

A27.6

ABB

Stress: 0,18

DHABs Diversity (families)nMDS analysisnMDS analysis

Diversity indexes (families)Diversity indexes (families)

Urania

0

0,2

0,4

0,6

0,8

1

U3.8 U5.1 U8.2 U8.7 U10.9 U14 U14.6 U15.6 UBB

Sample(%salinity)

Dom

inan

ce

0

0,5

1

1,5

2

2,5

Div

ersi

ty

(Sha

nnon

)

Dominance

Shannon Bannock

0

0,2

0,4

0,6

0,8

1

B3.9 B12.8 B14.4 B16.9 B22.4 B24.6 B28

Sample(%salinity)

Dom

inan

ce

0

0,5

1

1,5

2

2,5

Div

ersi

ty

(Sha

nnon

)

Dominance

Shannon

L'Atalante

0

0,2

0,4

0,6

0,8

1

A4 A7,8 A12,8 A19,4 A23,8 A27.6 ABB

Sample(%salinity)

Dom

inan

ce

0

0,5

1

1,5

2

2,5D

iver

sity

(S

hann

on)

Dominance

ShannonDiscovery

0

0,2

0,4

0,6

0,8

1

D6.2 D9 D15.4 D17* D21.2 D24.4 D26* DBB

Sample(%salinity)

Dom

inan

ce

0

0,5

1

1,5

2

2,5

Div

ersi

ty

(Sha

nnon

)

Dominance

Shannon

High diversity – Low dominance Low diversity, High dominance

7. Urania show a regular increasing of diversity along the gradient

1. At level of bacterial Family every basin seems to be a different world!Family every basin seems to be a different world!

3. Urania, Bannock, and L’Atalante shows the highest diversity

4. Urania, Bannock, and L’Atalante shows the lowest dominance

5. Discovery shows lower level of diversity

6. Discovery shows dominance of specific families at medium salinity concentration along the gradient

2. There is a grouping made by SW and BB layer………. “Real o Bias?”

U3.8

U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6

UB

B

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2

D9

D15.4

D17*

D21.2

D24.4

D26*

DB

B

A4

A7,8

A12,8

A19,4

A23,8

A27.6

AB

B

ArcheonDHABs [CD1]Thermotog.CFBActinobacteriaCandidate division sed 3KB1VerrucomicrobialesClostridialesFlexibact.DHABs[CD2]DHABs[CD3]Spirochaet.Acidobact.Rhizob.Rhodobact.Magnetic BacteriumDHABs[CD4]Desulfuromon.Desulfobact.Desulfoarc.Geobact.Desulfohalob.Desulfobulb.Syntrophobact.SB1UnculturedHelicobact.

Campylobact.Moraxellac.Psudomonad.OceanospirillalesEnterobact.Pisciricketts.Alteromon.UnculturedGDechlorimarinusEctothiorhodospir.Chromat.Alcanivorax.

Urania Bannock Discovery L’Atalante

U3.8

U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6

UB

B

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2

D9

D15.4

D17*

D21.2

D24.4

D26*

DB

B

A4

A7,8

A12,8

A19,4

A23,8

A27.6

AB

B

UraniaUrania

ArcheonDHABs [CD1]Thermotog.CFBActinobacteriaCandidate division sed 3KB1VerrucomicrobialesClostridialesFlexibact.DHABs[CD2]DHABs[CD3]Spirochaet.Acidobact.Rhizob.Rhodobact.Magnetic BacteriumDHABs[CD4]Desulfuromon.Desulfobact.Desulfoarc.Geobact.Desulfohalob.Desulfobulb.Syntrophobact.SB1UnculturedHelicobact.

Campylobact.Moraxellac.Psudomonad.OceanospirillalesEnterobact.Pisciricketts.Alteromon.UnculturedGDechlorimarinusEctothiorhodospir.Chromat.Alcanivorax.

Archeon Flexibact. 0.793Archeon Pisciricketts. 0.758DHABs [CD1] Helicobact. 0.886DHABs [CD1] Pisciricketts. 0.811Thermotog. KB1 1

Actinobacteria Moraxellac. 1Actinobacteria Enterobact. 0.834Cand. Div. sed 3 DHABs[CD3] 1Cand. Div. sed 3 Spirochaet. 1Cand. Div. sed 3 Magnetic Bacterium 1Cand. Div. sed 3 Desulfohalob. 0.837KB1 Acidobact. 0.796Clostridiales Rhizob. 1Clostridiales Geobact. 0.735Clostridiales Ectothiorhodospir. 1DHABs[CD3] Spirochaet. 1DHABs[CD3] Magnetic Bacterium 1DHABs[CD3] Desulfohalob. 0.837Spirochaet. Magnetic Bacterium 1Spirochaet. Desulfohalob. 0.837Acidobact. Desulfobact. 0.766Acidobact. Desulfobulb. 0.785Rhizob. Geobact. 0.735Rhizob. Ectothiorhodospir. 1Magnetic BacteriumDesulfohalob. 0.837Geobact. Ectothiorhodospir. 0.735Helicobact. Campylobact. 0.736Helicobact. Pisciricketts. 0.782Moraxellac. Enterobact. 0.834Psudomonad. Enterobact. 0.977

Direct and Inverse Correlations (Pearson’s test, P<0.05 (uncorr))

U3.8

U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6

UB

B

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2

D9

D15.4

D17*

D21.2

D24.4

D26*

DB

B

A4

A7,8

A12,8

A19,4

A23,8

A27.6

AB

B

BannockBannock

Archeon Desulfobulb. 0,983DHABs [CD1] Desulfobact. 0,794CFB Desulfoarc. 1CFB UnculturedG 1Actinobacteria Pisciricketts. 0,735Cand. Div. sed 3 Clostridiales 0,936Cand. Div. sed 3 DHABs[CD2] 0,807KB1 Verrucomicrobiales 0,99KB1 DHABs[CD3] 0,938

KB1 SB1 0,839Verrucomicrobiales DHABs[CD3] 0,879

Verrucomicrobiales SB1 0,79Clostridiales Acidobact. 0,729Clostridiales Geobact. 0,729Flexibact. Moraxellac. 1Flexibact. Psudomonad. 0,821DHABs[CD3] Desulfohalob. 0,704DHABs[CD3] SB1 0,888Acidobact. Geobact. 1DHABs[CD3] Enterobact. -0,701Desulfoarc. UnculturedG 1Desulfohalob. Alteromon. 0,76Moraxellac. Psudomonad. 0,821SB1 Psudomonad. -0,74

Direct and Inverse Correlations (Pearson’s test, P<0.05 (uncorr))ArcheonDHABs [CD1]Thermotog.CFBActinobacteriaCandidate division sed 3KB1VerrucomicrobialesClostridialesFlexibact.DHABs[CD2]DHABs[CD3]Spirochaet.Acidobact.Rhizob.Rhodobact.Magnetic BacteriumDHABs[CD4]Desulfuromon.Desulfobact.Desulfoarc.Geobact.Desulfohalob.Desulfobulb.Syntrophobact.SB1UnculturedHelicobact.

Campylobact.Moraxellac.Psudomonad.OceanospirillalesEnterobact.Pisciricketts.Alteromon.UnculturedGDechlorimarinusEctothiorhodospir.Chromat.Alcanivorax.

1

U3.8

U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6

UB

B

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2

D9

D15.4

D17*

D21.2

D24.4

D26*

DB

B

A4

A7,8

A12,8

A19,4

A23,8

A27.6

AB

B

DiscoveryDiscovery

Archeon

Clostridiales

1Archeon

Spirochaet. 1Archeon

Dechlorimarinus

0,844CFB

Acidobact.

0,722CFB

Ectothiorhodospir. 0,896KB1

Verrucomicrobiales

0,851Clostridiales

Spirochaet. 1Clostridiales

Dechlorimarinus

0,844Flexibact.

Rhodobact. 1Flexibact.

Desulfobulb.

1DHABs[CD2]

UnculturedG 0,755DHABs[CD2]

Chromat. 1Spirochaet. Dechlorimarinu

s0,844

Rhodobact. Desulfobulb.

1DHABs[CD4]

Desulfuromon. 0,751DHABs[CD4]

Ectothiorhodospir. 0,722Desulfobact.

SB1

0,761Desulfobact.

Alcanivorax.

0,736SB1

Alcanivorax.

0,777Psudomonad.

Alteromon. 0,889UnculturedG Chromat. 0,755Dechlorimarinus

Alcanivorax.

0,84

Direct and Inverse Correlations (Pearson’s test, P<0.05 (uncorr))ArcheonDHABs [CD1]Thermotog.CFBActinobacteriaCandidate division sed 3KB1VerrucomicrobialesClostridialesFlexibact.DHABs[CD2]DHABs[CD3]Spirochaet.Acidobact.Rhizob.Rhodobact.Magnetic BacteriumDHABs[CD4]Desulfuromon.Desulfobact.Desulfoarc.Geobact.Desulfohalob.Desulfobulb.Syntrophobact.SB1UnculturedHelicobact.

Campylobact.Moraxellac.Psudomonad.OceanospirillalesEnterobact.Pisciricketts.Alteromon.UnculturedGDechlorimarinusEctothiorhodospir.Chromat.Alcanivorax.

2

U3.8

U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6

UB

B

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2

D9

D15.4

D17*

D21.2

D24.4

D26*

DB

B

A4

A7,8

A12,8

A19,4

A23,8

A27.6

AB

B

L’AtalanteL’Atalante

DHABs [CD1] DHABs[CD2] 1

KB1 Pisciricketts. -0,725

Rhodobact. Campylobact. 0,766

Campylobact. Pisciricketts. 0,828

Oceanospirillales Alcanivorax. 1

Direct and Inverse Correlations (Pearson’s test, P<0.05 (uncorr))ArcheonDHABs [CD1]Thermotog.CFBActinobacteriaCandidate division sed 3KB1VerrucomicrobialesClostridialesFlexibact.DHABs[CD2]DHABs[CD3]Spirochaet.Acidobact.Rhizob.Rhodobact.Magnetic BacteriumDHABs[CD4]Desulfuromon.Desulfobact.Desulfoarc.Geobact.Desulfohalob.Desulfobulb.Syntrophobact.SB1UnculturedHelicobact.

Campylobact.Moraxellac.Psudomonad.OceanospirillalesEnterobact.Pisciricketts.Alteromon.UnculturedGDechlorimarinusEctothiorhodospir.Chromat.Alcanivorax.

1. There are a specific DHABs Configuration

2. There are some unique and sporadic phylotypes (probably “contamination”?)

3. There are Specific direct correlations within tipical DHABs flora.

4. Gamma proteobacteria shows to be often inverse related with typical DHABs Flora

5. Discovery Basin show a peculiar configuration with two big groups(KB1 and Desulfobatcereaceae) domainig two different layer of the basin interface

6. In Discovery and Bannock Basin KB1 and Verrucomicrobiales go very well related along the basin interface gradient

8. In Atalante, there are very well defined bacterial distributions

Urania

Bannock

Discovery

L'Atalante

Stress: 0,16

2nd Statistical aproach: Divisions Level2nd Statistical aproach: Divisions Level

nMDS analysisnMDS analysis DHABs Groups (Divisons)

Urania

Bannock

Discovery

L'Atalante

Stress: 0,16

U14

U14.6

UBB

B3.9B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2D9

DBB

A4

A7,8A12,8

A19,4A23,8

ABB

U3.8

nMDS analysisnMDS analysis DHABs Groups (Divisons)

Urania

Bannock

Discovery

L'Atalante

Stress: 0,16

U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6UBB

B3.9B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2D9

DBB

A4

A7,8A12,8

A19,4A23,8

ABB

U3.8

nMDS analysisnMDS analysis DHABs Groups (Divisons)

DHABs Groups (Divisons)

Urania

Bannock

Discovery

L'Atalante

Stress: 0,16

U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6UBB

B3.9B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2D9

D15.4

D17*D21.2

D24.4D26*

DBB

A4

A7,8A12,8

A19,4A23,8

A27.6

ABB

U3.8

nMDS analysisnMDS analysis

At level of functional group:functional group:

1. There is a common significance that join the more saline Urania layers with the Bannock and Atalante basins

2. Discovery maintains its peculiar bizarre world

3. Some layers of Urania seems to drop out from the common distribution

DHABs [CD1]

U3.8U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6UBB

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2D9

D15.4

D17*D21.2

D24.4D26*

DBB

A4

A7,8A12,8

A19,4A23,8

A27.6

ABB

Stress: 0,16

nMDS analysisnMDS analysis

DHABs CFB

U3.8U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6UBB

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2D9

D15.4

D17*D21.2

D24.4D26*

DBB

A4

A7,8A12,8

A19,4A23,8

A27.6

ABB

Stress: 0,16

nMDS analysisnMDS analysis

DHABs Actinobactereaceae

U3.8U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6UBB

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2D9

D15.4

D17*D21.2

D24.4D26*

DBB

A4

A7,8A12,8

A19,4A23,8

A27.6

ABB

Stress: 0,16

nMDS analysisnMDS analysis

DHABs Candidate Division Sediment 3

U3.8U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6UBB

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2D9

D15.4

D17*D21.2

D24.4D26*

DBB

A4

A7,8A12,8

A19,4A23,8

A27.6

ABB

Stress: 0,16

nMDS analysisnMDS analysis

DHABs KB1

U3.8U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6UBB

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2D9

D15.4

D17*D21.2

D24.4D26*

DBB

A4

A7,8A12,8

A19,4A23,8

A27.6

ABB

Stress: 0,16

nMDS analysisnMDS analysis

DHABs Verrucomicrobiales

U3.8U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6UBB

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2D9

D15.4

D17*D21.2

D24.4D26*

DBB

A4

A7,8A12,8

A19,4A23,8

A27.6

ABB

Stress: 0,16

nMDS analysisnMDS analysis

DHABs [CD2]

U3.8U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6UBB

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2D9

D15.4

D17*D21.2

D24.4D26*

DBB

A4

A7,8A12,8

A19,4A23,8

A27.6

ABB

Stress: 0,16

nMDS analysisnMDS analysis

DHABs [CD3]

U3.8U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6UBB

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2D9

D15.4

D17*D21.2

D24.4D26*

DBB

A4

A7,8A12,8

A19,4A23,8

A27.6

ABB

Stress: 0,16

nMDS analysisnMDS analysis

DHABs Spiro

U3.8U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6UBB

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2D9

D15.4

D17*D21.2

D24.4D26*

DBB

A4

A7,8A12,8

A19,4A23,8

A27.6

ABB

Stress: 0,16

nMDS analysisnMDS analysis

DHABs Acidobactereaceae

U3.8U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6UBB

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2D9

D15.4

D17*D21.2

D24.4D26*

DBB

A4

A7,8A12,8

A19,4A23,8

A27.6

ABB

Stress: 0,16

nMDS analysisnMDS analysis

DHABs [CD4]

U3.8U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6UBB

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2D9

D15.4

D17*D21.2

D24.4D26*

DBB

A4

A7,8A12,8

A19,4A23,8

A27.6

ABB

Stress: 0,16

nMDS analysisnMDS analysis

DHABs Alpha Proteobacteria

U3.8U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6UBB

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2D9

D15.4

D17*D21.2

D24.4D26*

DBB

A4

A7,8A12,8

A19,4A23,8

A27.6

ABB

Stress: 0,16

nMDS analysisnMDS analysis

DHABs Delta Proteobacteria

U3.8U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6UBB

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2D9

D15.4

D17*D21.2

D24.4D26*

DBB

A4

A7,8A12,8

A19,4A23,8

A27.6

ABB

Stress: 0,16

nMDS analysisnMDS analysis

DHABs Epsilon Proteobacteria

U3.8U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6UBB

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2D9

D15.4

D17*D21.2

D24.4D26*

DBB

A4

A7,8A12,8

A19,4A23,8

A27.6

ABB

Stress: 0,16

nMDS analysisnMDS analysis

DHABs SB1

U3.8U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6UBB

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2D9

D15.4

D17*D21.2

D24.4D26*

DBB

A4

A7,8A12,8

A19,4A23,8

A27.6

ABB

Stress: 0,16

nMDS analysisnMDS analysis

DHABs Gamma Proteobacteria

U3.8U5.1

U8.2

U8.7

U10.9

U14

U14.6

U15.6UBB

B3.9

B12.8

B14.4

B16.9

B22.4

B24.6

B28

D6.2D9

D15.4

D17*D21.2

D24.4D26*

DBB

A4

A7,8A12,8

A19,4A23,8

A27.6

ABB

Stress: 0,16

nMDS analysisnMDS analysis

> Gamma [] [] []

Epsilon

Desulfobac>Desulfobulb

Archaea–Desulfobact (D) GeobactereaceaeHIGH DIVERSITY ZONE

KB1+Verr>Desulfobact. (D)Desulfohalob> Desulfobact

Desulfobulb>Desulfobact(B)

KB1+Verr (D)

> Gamma [] [] []

Proposed metabolic pathways in the DHABsProposed metabolic pathways in the DHABs

3380dbar

e-

e-

e-

e-

e-

e-

e-

e-

e-

Volatile fatty acids

CO2

MnMn++++ BaBa++++ SulphateSulphate SulphideSulphide SulphurSulphur

Oxic

Anoxic

Acetate

RedOx Potential

NONO33

MnOMnO22

(FeOH)(FeOH)33

SOSO442-2-

COCO22

OO22

Eh:-232 -313

* Evidence of in situ active cells or functional genes involved in the pathways

e-e-

CO2

CH4

Methanogens *AOM andMethanohalophilus 16SrRNA

SO42-

S-

S0SRB

e-

e-*dsrAB func.genes

Fe(OH)3

Fe2+

FeRBe- *

Geobacteriaceae 16SrRNA

MnOMnO22

MnMn2+2+

BaBa++++

BaBa++++Manganese oxidisers

*

Arcobacter 16SrRNA

Discovery Functional charachterisationDiscovery Functional charachterisation

In study: Distribution along the Discovery salinity graident of:

Anaerobic Methane Oxidation related (ANME) bacteria (rRNA)

Desulfobatcteraceae distribution (16rRNA)

DSR Subunit A gene (RNA)DSR Subunit B gene (RNA)

0.1

Methanocaldococcus sp.

Methanococcus voltae Methanococcus aeolicus

Methanothermococcus okinawensis

Halogeometricum borinquense Halophilic archaeon PalaeII

AMO-11%-B24

Unidentified archaeon

AMO11%-10AMO11%-5AMO11%-9

Deep Hidrotermal Vent Environment

Group 3

Uncultured archaeon SAGMA-10

Unidentified Crenoarchaeota

Uncultured Crenoarchaeota

Uncultured archaeon 19a-14

A20-11%-B1A20-6%-27

A20-11%-B2A20-6%-12A20-6%-8A20-6%-29

Methanohalophilus euhalobius

Methanohalophilus portucalensis

AMO-19%-6AMO-19%-7AMO-11%-2AMO-11%-3A20-11%-19A20-11%-11AMO-19%-5AMO-11%-1

ANME-2c

Halophilic Metanogens

and ANME

Discovery Anaerobic Methane Oxidation related (ANME) - bacteria (16SrRNA)Discovery Anaerobic Methane Oxidation related (ANME) - bacteria (16SrRNA)

0.1

6%-226%-3

Desulfobacterium phenolicum

Uncultured clone DBBB-72

Desulfobacula toluolica

6%-7

6%-56%-36

Uncultured clone DLIB-25

11%-31

6%-35

Desulfobacterium vacuolatum

11%-20

Desulfotignum phosphitoxidans

6%-19

Desulfobacula sp.

Desulfospira joergensenii

Desulfobacula toluolica11%-35

6%-216%-26

SRB clones at 6% salinity

SRB clones at 11% salinity

Discovery Desulfobactereaceae distribution clones - bacteria (16SrRNA)Discovery Desulfobactereaceae distribution clones - bacteria (16SrRNA)

0.1

Guaymas basin clone B01P012Guaymas basin clone B01P020

Desulfobacula toluolica

Desulfotignum balticum

Archaeoglobus fulgidus M95624

Thermodesulfovibrio islandicus

Desulfobacter latus U58124

Desulfotomaculum thermobenzoicum

Desulfoarculus baarsii

Desulfovibrio vulgarisDesulfovibrio desulfuricans

Desulfotomaculum alkaliphilum Desulfotomaculum putei

Pelotomaculum sp. MGP

Desulfosporosinus orientensis

KYF_135

Thermodesulforhabdus norvegica

Desulforhabdus amnigena

Thermodesulfobacterium commune

Guaymas basin clone B04P001

Desulfococcus multivorans

Desulforhopalus vacuolatus

Desulfobacterium vacuolatum

11%-54

11%-5111%-82

Discovery basin SRB clones 6-11%

Dissimilatory sulfite reductase Alpha subunit (dsrA)

11%-82

SRB clones at 6% salinity

SRB clones at 11% salinity

0.1

Thermodesulfovibrio islandicus

Thermodesulfovibrioyellowstonii

Thermodesulforhabdus norvegica

Pelotomaculum sp. MGP

Desulforhabdus amnigena

Thermodesulfobacterium commune

Desulfomonile tiedjei

Desulfotomaculum thermobenzoicum

Guaymas basin clone B04P001

Desulfobacterium vacuolatum

Desulfotignum balticum11%-51

Desulfobacula toluolica

B01P020

11%-77 Desulforhopalus vacuolatusDesulforhopalus singaporensis

Desulfovibrio desulfuricansDesulfovibrio vulgaris

Desulfoarculus baarsii

Desulfococcus multivorans

Desulfotomaculum putei

Discovery basinSRB clones 6-11%

SRB clones at 6% salinity

SRB clones at 11% salinity

Dissimilatory sulfite reductase Beta subunit (dsrB)

1. At 6% of salinity of Discovery basin we found only members related to crenoarcheota division

2. At 11% and 19% the new DHVE3

3. The anaerobic methanophiles was found at 11% and 19% of salinity

- Bacteria

4. 16SrRNA from 6% and 11% of salinity shows mainly Desulfobacteraceae related bacteria

5. dsrA and dsrB distribution shows two levels of diversity (macro and micro)

6. the dsr clusters related to the Desulfobacteriaceae show an high ratio of microdiversity with two defined sub-clusters:

i) 6% of salinity clones ii) 6% and 11% togeter

- Archaea