Presentazione di PowerPoint

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CNR IAMC - Istituto per lAmbiente Marino
Costiero Messina, Italy Università degli Studi
di Messina
WP3
State of the Art May 2004
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16 rRNA Distribution at Family level In study
gt830 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 LAtalante from 4 of salinity to
BB 1st Statistical aproach Family level
(-aceae) 2nd Statistical aproach Division
Level
3
16S rRNA Bannock Sampling Profile
Urania
Bannock
Eh
Eh
-350
-250
-150
-50
50
-300
-250
-200
-150
-100
-50
0
50
0
1,338
0
1,335
1,34
1,340
1,342
5
3,8
4,2
1,345
3,9
5
6
5,1
1,344
1,350
6,7
9,8
10
10,2
1,346
1,355
11
7,7
8,2
12,8
8,7
1,360
9
1,348
14,4
15
10
15,4
1,365
10,9
10,9
1,35
16,9
19
1,370
1,352
20
20,6
13,4
1,375
13,9
22.4
14,6
1,354
23,2
15
15
15,3
1,380
15,6
24.6
25
16
1,356
1,385
28
1,358
30
1,390
20
1,36
LAtalante
Discovery
Eh
Eh
-350
-300
-250
-200
-150
-100
-50
0
-300
-250
-200
-150
-100
-50
0
50
0
1,335
0
1,34
1,34
4
1,35
4,4
4,8
5
5,2
5
1,345
6,2
7
7
7,8
1,36
1,35
9
10
10
1,355
12,8
13
1,37
15
1,36
15
15,4
1,38
1,365
18
19,4
20
20
1,37
21,2
21,2
1,39
23,8
1,375
24
24,2
24,4
25
25
25,4
25,6
1,4
26
1,38
27,6
30
1,385
30
1,41
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nMDS analysis
DHABs Diversity (families)
Stress 0,18
Urania
Bannock
Discovery
L'Atalante
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nMDS analysis
DHABs Diversity (families)
Stress 0,18
A4
Urania
U3.8
B28
UBB
DBB
U5.1
U8.7
B3.9
Bannock
U15.6
B12.8
U8.2
U10.9
B16.9
A12,8
A7,8
U14.6
U14
B24.6
B14.4
Discovery
A23,8
A19,4
B22.4
D9
D24.4
A27.6
D21.2
ABB
D15.4
L'Atalante
D17
D26
D6.2
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Diversity indexes (families)
High diversity Low dominance
Low diversity, High dominance
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1. At level of bacterial Family every basin seems
to be a different world!
2. There is a grouping made by SW and BB
layer. Real o Bias?
3. Urania, Bannock, and LAtalante shows the
highest diversity
4. Urania, Bannock, and LAtalante 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
7. Urania show a regular increasing of diversity
along the gradient
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Urania
Bannock
Discovery
LAtalante
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Urania
Direct and Inverse Correlations (Pearsons test,
Plt0.05 (uncorr))
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Urania
Bannock
Direct and Inverse Correlations (Pearsons test,
Plt0.05 (uncorr))
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Discovery
Direct and Inverse Correlations (Pearsons test,
Plt0.05 (uncorr))
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LAtalante
Direct and Inverse Correlations (Pearsons test,
Plt0.05 (uncorr))
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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
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nMDS analysis
DHABs Groups (Divisons)
Stress 0,16
Urania
Bannock
Discovery
L'Atalante
2nd Statistical aproach Divisions Level
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nMDS analysis
DHABs Groups (Divisons)
Stress 0,16
A4
U3.8
Urania
UBB
DBB
A12,8
B28
A7,8
B3.9
B12.8
Bannock
B16.9
D6.2
D9
ABB
A19,4
U14.6
A23,8
B22.4
Discovery
B14.4
U14
B24.6
L'Atalante
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nMDS analysis
DHABs Groups (Divisons)
Stress 0,16
U5.1
A4
U3.8
Urania
UBB
DBB
U15.6
A12,8
B28
A7,8
U8.7
B3.9
B12.8
U8.2
Bannock
U10.9
B16.9
D6.2
D9
ABB
A19,4
U14.6
A23,8
B22.4
Discovery
B14.4
U14
B24.6
L'Atalante
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nMDS analysis
DHABs Groups (Divisons)
Stress 0,16
Urania
Bannock
Discovery
L'Atalante
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At level of 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
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nMDS analysis
DHABs CD1
Stress 0,16
U3.8
U5.1
A4
UBB
DBB
U15.6
A12,8
B28
A7,8
U8.7
B3.9
B12.8
U8.2
U10.9
B16.9
D6.2
D9
ABB
A19,4
U14.6
A23,8
B22.4
B14.4
U14
D15.4
D26
B24.6
D24.4
A27.6
D17
D21.2
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nMDS analysis
DHABs CFB
Stress 0,16
U3.8
U5.1
A4
UBB
DBB
U15.6
A12,8
B28
A7,8
U8.7
B3.9
B12.8
U8.2
U10.9
B16.9
D6.2
D9
ABB
A19,4
U14.6
A23,8
B22.4
B14.4
U14
D15.4
D26
B24.6
D24.4
A27.6
D17
D21.2
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nMDS analysis
DHABs Actinobactereaceae
Stress 0,16
U3.8
U5.1
A4
UBB
DBB
U15.6
A12,8
B28
A7,8
U8.7
B3.9
B12.8
U8.2
U10.9
B16.9
D6.2
D9
ABB
A19,4
U14.6
A23,8
B22.4
B14.4
U14
D15.4
D26
B24.6
D24.4
A27.6
D17
D21.2
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nMDS analysis
DHABs Candidate Division Sediment 3
Stress 0,16
U3.8
U5.1
A4
UBB
DBB
U15.6
A12,8
B28
A7,8
U8.7
B3.9
B12.8
U8.2
U10.9
B16.9
D6.2
D9
ABB
A19,4
U14.6
A23,8
B22.4
B14.4
U14
D15.4
D26
B24.6
D24.4
A27.6
D17
D21.2
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nMDS analysis
DHABs KB1
Stress 0,16
U3.8
U5.1
A4
UBB
DBB
U15.6
A12,8
B28
A7,8
U8.7
B3.9
B12.8
U8.2
U10.9
B16.9
D6.2
D9
ABB
A19,4
U14.6
A23,8
B22.4
B14.4
U14
D15.4
D26
B24.6
D24.4
A27.6
D17
D21.2
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nMDS analysis
DHABs Verrucomicrobiales
Stress 0,16
U3.8
U5.1
A4
UBB
DBB
U15.6
A12,8
B28
A7,8
U8.7
B3.9
B12.8
U8.2
U10.9
B16.9
D6.2
D9
ABB
A19,4
U14.6
A23,8
B22.4
B14.4
U14
D15.4
D26
B24.6
D24.4
A27.6
D17
D21.2
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nMDS analysis
DHABs CD2
Stress 0,16
U3.8
U5.1
A4
UBB
DBB
U15.6
A12,8
B28
A7,8
U8.7
B3.9
B12.8
U8.2
U10.9
B16.9
D6.2
D9
ABB
A19,4
U14.6
A23,8
B22.4
B14.4
U14
D15.4
D26
B24.6
D24.4
A27.6
D17
D21.2
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nMDS analysis
DHABs CD3
Stress 0,16
U3.8
U5.1
A4
UBB
DBB
U15.6
A12,8
B28
A7,8
U8.7
B3.9
B12.8
U8.2
U10.9
B16.9
D6.2
D9
ABB
A19,4
U14.6
A23,8
B22.4
B14.4
U14
D15.4
D26
B24.6
D24.4
A27.6
D17
D21.2
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nMDS analysis
DHABs Spiro
Stress 0,16
U3.8
U5.1
A4
UBB
DBB
U15.6
A12,8
B28
A7,8
U8.7
B3.9
B12.8
U8.2
U10.9
B16.9
D6.2
D9
ABB
A19,4
U14.6
A23,8
B22.4
B14.4
U14
D15.4
D26
B24.6
D24.4
A27.6
D17
D21.2
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nMDS analysis
DHABs Acidobactereaceae
Stress 0,16
U3.8
U5.1
A4
UBB
DBB
U15.6
A12,8
B28
A7,8
U8.7
B3.9
B12.8
U8.2
U10.9
B16.9
D6.2
D9
ABB
A19,4
U14.6
A23,8
B22.4
B14.4
U14
D15.4
D26
B24.6
D24.4
A27.6
D17
D21.2
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nMDS analysis
DHABs CD4
Stress 0,16
U3.8
U5.1
A4
UBB
DBB
U15.6
A12,8
B28
A7,8
U8.7
B3.9
B12.8
U8.2
U10.9
B16.9
D6.2
D9
ABB
A19,4
U14.6
A23,8
B22.4
B14.4
U14
D15.4
D26
B24.6
D24.4
A27.6
D17
D21.2
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nMDS analysis
DHABs Alpha Proteobacteria
Stress 0,16
U3.8
U5.1
A4
UBB
DBB
U15.6
A12,8
B28
A7,8
U8.7
B3.9
B12.8
U8.2
U10.9
B16.9
D6.2
D9
ABB
A19,4
U14.6
A23,8
B22.4
B14.4
U14
D15.4
D26
B24.6
D24.4
A27.6
D17
D21.2
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nMDS analysis
DHABs Delta Proteobacteria
Stress 0,16
U3.8
U5.1
A4
UBB
DBB
U15.6
A12,8
B28
A7,8
U8.7
B3.9
B12.8
U8.2
U10.9
B16.9
D6.2
D9
ABB
A19,4
U14.6
A23,8
B22.4
B14.4
U14
D15.4
D26
B24.6
D24.4
A27.6
D17
D21.2
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nMDS analysis
DHABs Epsilon Proteobacteria
Stress 0,16
U3.8
U5.1
A4
UBB
DBB
U15.6
A12,8
B28
A7,8
U8.7
B3.9
B12.8
U8.2
U10.9
B16.9
D6.2
D9
ABB
A19,4
U14.6
A23,8
B22.4
B14.4
U14
D15.4
D26
B24.6
D24.4
A27.6
D17
D21.2
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nMDS analysis
DHABs SB1
Stress 0,16
U3.8
U5.1
A4
UBB
DBB
U15.6
A12,8
B28
A7,8
U8.7
B3.9
B12.8
U8.2
U10.9
B16.9
D6.2
D9
ABB
A19,4
U14.6
A23,8
B22.4
B14.4
U14
D15.4
D26
B24.6
D24.4
A27.6
D17
D21.2
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nMDS analysis
DHABs Gamma Proteobacteria
Stress 0,16
U3.8
U5.1
A4
UBB
DBB
U15.6
A12,8
B28
A7,8
U8.7
B3.9
B12.8
U8.2
U10.9
B16.9
D6.2
D9
ABB
A19,4
U14.6
A23,8
B22.4
B14.4
U14
D15.4
D26
B24.6
D24.4
A27.6
D17
D21.2
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gt Gamma Epsilon DesulfobacgtDesulfobulb
ArchaeaDesulfobact (D) Geobactereaceae HIGH
DIVERSITY ZONE KB1VerrgtDesulfobact.
(D) Desulfohalobgt Desulfobact DesulfobulbgtDesulfo
bact(B) KB1Verr (D) gt Gamma
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Proposed metabolic pathways in the DHABs
O2
Oxic
3380dbar
NO3
Anoxic
MnO2
Volatile fatty acids
(FeOH)3
Acetate
e-
SO42-
e-
e-
e-
CO2
e-
e-
CO2
e-
e-
e-
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Discovery 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)
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Discovery Anaerobic Methane Oxidation related
(ANME) - bacteria (16SrRNA)
Methanocaldococcus
sp.
Methanococcus voltae
Methanococcus aeolicus
Methanothermococcus okinawensis
0.1
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Discovery Desulfobactereaceae distribution clones
- bacteria (16SrRNA)
6-19
Desulfobacula sp.
Desulfotignum phosphitoxidans
Desulfospira joergensenii
Desulfobacula toluolica
Desulfobacterium vacuolatum
11-35
6-26
6-22
6-21
6-3
6-35
11-20
Desulfobacterium phenolicum
11-31
Uncultured clone DLIB-25
6-5
Uncultured clone DBBB-72
6-36
0.1
Desulfobacula toluolica
6-7
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Discovery basin SRB clones 6-11
Dissimilatory sulfite reductase Alpha subunit
(dsrA)
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Discovery basin
SRB clones 6-11
Dissimilatory sulfite reductase Beta subunit
(dsrB)
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- Archaea
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