PALEOSALT 2008 Bivalves - VUB Rémy Mas, Fabrice Servaes, Maité Bauwens, Céline Poulain, Philippe...

PALEOSALT 2008Bivalves - VUB

Rémy Mas, Fabrice Servaes, Maité Bauwens, Céline Poulain, Philippe

Claeys, Eddy Keppens, Frank Dehairs

Objectives

• Understanding impact of external controls (environmental conditions) on Trace Elements and Isotopes incorporation in bivalve shells

Aquarium experiment

• Development of the δD salinity proxy applied to organic matter in bivalve shells

In situ work : Scheldt estuary and Auray river

Aquarium Experiments 1

• Sampling– 05 Feb 07– Mytilus edulis– knokke

• Acclimation• Same temperature and salinity as used for

experiment

• Experiment– Start 19 Feb 07– End 30 April 07

Knokke

T1

T2

S1

S2

S1

S2

F2

F2

F1

F2

F1

F2

T1 S2 F2Adult

Juvenile


– Temperature• T18°C ; T216°C

– Salinity• S228 (sea water)δ18Omean = -0.31 ‰

δ13CDIC = -0,36 ‰

• S118 (diluted water)δ18Omean = -2.85 ‰

δ13CDIC = -0,37 ‰

- Food : Tetracelmis suecica; δ13C=-30‰

F1=8mgDW/week/org.F2=16mgDW/week/org.

- Age : Juvenile (<1year) Vs ‘Adult’ (~1 year old)

Aquarium Experiments 3• Growing (∆L of the shell)

Difference statistically very significant

Difference statistically significant

Difference statistically not significant

dL shell Student test Count Average p

Temperature

T1S1F2 Vs T2S1F2 6 10 1.53 2.230.0001

2

T1S2F2 Vs T2S2F2 8 10 2.00 2.92 3.4E-06

Salinity

T1S1F2 Vs T1S2F2 6 8 1.53 2.000.0202

7

T2S1F1 Vs T2S2F1 10 10 2.24 2.820.0005

7

T2S1F2 Vs T2S2F2 10 10 2.23 2.92 2.1E-06

FoodT2S1F2 Vs T2S1F1 10 10 2.23 2.24

0.94732

T2S2F2 Vs T2S2F1 10 10 2.92 2.820.2553

5

AgeAT1S2F2 Vs

T1S2F2 7 8 1.69 1.460.0698

5Temperature ∆L Salinity ∆L


• C isotopic composition of mussel tissues

d13C tissues Student test Count Average p

Temperature

T1S1F2 Vs T2S1F2 14 15 -22.07 -22.070.9887

7

T1S2F2 Vs T2S2F2 17 17 -21.92 -22.080.3963

3

Salinity

T1S1F2 Vs T1S2F2 14 17 -22.07 -21.920.4320

7

T2S1F1 Vs T2S2F1 14 19 -21.92 -21.930.9068

2

T2S1F2 Vs T2S2F2 15 17 -22.07 -22.080.9576

1

Food T2S1F2 Vs T2S1F1 15 14 -22.07 -21.920.1973

7

T2S2F2 Vs T2S2F1 17 19 -22.08 -21.93 0.3391

AgeAT1S2F2 Vs T1S2F2 14 17 -22.23 -21.92

0.08727




d15N tissues Student test Count Average p

TemperatureT1S1F2 Vs T2S1F2 14 17 7.33 9.36 0.02165

T1S2F2 Vs T2S2F2 17 17 7.86 7.15 0.54666

Salinity

T1S1F2 Vs T1S2F2 14 17 7.33 7.86 0.66162

T2S1F1 Vs T2S2F1 14 19 8.94 7.19 0.13884

T2S1F2 Vs T2S2F2 17 17 9.36 7.15 0.01496

FoodT2S1F2 Vs T2S1F1 17 14 9.36 8.94 0.45873

T2S2F2 Vs T2S2F1 17 19 7.15 7.19 0.97474

Age AT1S2F2 Vs T1S2F2 14 17 9.42 7.86 0.08854

• N isotopic composition of mussel tissues


• Carbonate– Sampling with microdrilling

1cm

300μm

800μm

Drill line

T1S2F2 01

-9-8-7-6-5-4-3-2-10

0 500 1000 1500 2000 2500 3000 3500

Distance from shell extremity (um)

d13

C

-1.8

-1.3

-0.8

-0.3

0.2

0.7

1.2

1.7

d18

O d13C

d18O

T1 Test 1

-9-8-7-6-5-4-3-2-10

0 500 1000 1500 2000 2500 3000 3500


d13

C

-1.8

-1.3

-0.8

-0.3

0.2

0.7

1.2

1.7

d18

O d13C

d18O

Adulteδ18Oss = 1.20 Tcalc= 9.5

Juvenileδ18Os =1.07

Tcalc = 10.0

T1=8°CS2=28

δ180w = -0.31

Wanamaker et al. 2007T(°C)=16.19-4.69*(δ18Os-δ18Ow)+0.17(δ18Os-δ18Ow)²

Adult

Juvenile

T2S2F2 01

-9-8-7-6-5-4-3-2-10

0 500 1000 1500 2000 2500 3000 3500


d13

C

-1.8

-1.3

-0.8

-0.3

0.2

0.7

1.2

1.7

d18

O d13C

d18O

T2S2F1 01

-9-8-7-6-5-4-3-2-10

0 500 1000 1500 2000 2500 3000 3500


d13

C

-1.8

-1.3

-0.8

-0.3

0.2

0.7

1.2

1.7

d18

O d13C

d18O

T2 S2 F2δ18Os = -0.23 Tcalc = 15.8

T2 S2 F1δ18Os = -0.54 Tcalc = 17.3

T2=16°CS2=28

δ18Ow = -0.31

T2S1F2 01

-9-8-7-6-5-4-3-2-10

0 500 1000 1500 2000 2500 3000 3500


d13

C

-1.8

-1.3

-0.8

-0.3

0.2

0.7

1.2

1.7

d18

O d13C

d18O

T2 S1 F2δ18Os = -0.17 Tcalc = 4.8

T2=16°CS1=18

δ18Ow = -2.85

Problem

T2S1F1 01

-9-8-7-6-5

-4-3-2-10

0 500 1000 1500 2000 2500 3000 3500 4000

distance (um)

d13

C

-1.8

-1.3

-0.8

-0.3

0.2

0.7

1.2

1.7

d13C

d18O

T2 S1 F1δ18Os = -0.33 Tcalc = 5.5


• Trace Elements (Ba, Sr, Mg, Mn…)– New machine : laser UP193FX(Fast

excimer) Merchantec 193nm. coupled to a ICP-MS Thermo X series 2. (Summer 2008)

Scheldt & Auray 1

• Sampling

LOCMARIAQUER

BONO

1 km

10 km

KN

HFGR

OS

Mytilus edulis

Ruditapesphilippinarum

0

5

10

15

20

25

30

35

2001-04 2002-09 2004-01 2005-05 2006-10 2008-02

Time

Sal

init

y

Breskens haven landzijde

Hansweert geul

Hoedekenskerke boei 4

Terneuzen boei 20

Vlissingen boei SSVH

Wielingen

KN

HF

GR

OS

Scheldt & Auray 2

Ruditapes philippinarumShell organic matrix

Mytilus edulis Shell organic matrix

Scheldt & Auray 3

Sea

Sea

• δ13C & δ15N from bivalves– Tissues– Shell OM

Scheldt & Auray 4

• Lipids– GC-MS

• Extraction of total lipids (modified Bligh & Dyer extraction) from bulk shell (without decarbonatation)

• Derivatization : Silylation BSTFA 60°C, 30min

• Method GC : – Column : DB-5 30m, Ø 0.25mm– 50°C (2min) 300°C (15min)

4°C/min

Mytilus edulis

Ruditapes

RT: 0.00 - 79.49

0 10 20 30 40 50 60 70Time (min)

0

50000000

100000000

150000000

200000000

250000000

300000000

350000000

400000000

450000000

500000000

550000000

600000000

Inte

nsi

ty

39.86

44.20

35.06

60.5164.68

50.139.22 54.30 67.729.58 34.1225.71 71.1217.537.38

NL:6.07E8TIC F: MS Me038KN

Fatty acids

Sterols

Me 038 KN

17:0

18:0

19:0

20:0

21:0

15:0

19

:1

Mytilus edulis

Scheldt & Auray 5

In the near future :• GC-MS

– Precise identification– Quantitative measurement

• GC-IRMS– δ13C measurements – Comparison of δ13C of specific

compounds with estuarine gradient

Perspectives

• Improvement of Mollusk shell Lipids extraction : Collaboration project with B. Farre; University Paris Sud; BIOCALC project; EUROCORES Program

• Deuterium analyses on specific lipidic compounds in collaboration with Dr. Stefan Schouten, Royal NIOZ

Thank you

C:\Xcalibur\...\blanc1_080606151011 6/6/2008 3:10:11 PM Me012GR

RT: 0.00 - 79.49

0 10 20 30 40 50 60 70Time (min)

0

10

20

30

40

50

60

70

80

90

100

Re

lativ

e A

bu

nd

an

ce

17.56

39.86

44.22

39.19

25.8535.06

16.289.17 64.6928.43 60.525.15 54.3212.43 18.83 67.7346.49 55.48 70.61

NL:5.88E8TIC F: MS blanc1_080606151011

R Time Intensity Formula Prob.

5.15 42855000 C8H18F3NOSi2 70.9 Acetamide, 2,2,2-trifluoro-N,N-bis(trimethylsilyl)-

9.17 58533636 C9H22OSi 29.7 Methoxydi(tert-butyl)silane

16.28 63595912 C6H14O2Si 27.2 Silanol, trimethyl-, propanoate

17.56 588324608 C10H26O3Si2 75.70 3,6,9-Trioxa-2,10-disilaundecane, 2,2,10,10-tetramethyl-

18.83 33368116 C12H32O3Si3 87.5 Trimethylsilyl ether of glycerol

22.58 31632980 C10H22N2O2Si2 70.9 2,4(1H,3H)-Pyrimidinedione, dihydro-1,3-bis(trimethylsilyl)-

25.85 101882128 C21H24O2 12 Benzene, 1,1'-(1-methylethylidene)bis[4-(2-propenyloxy)-

28.43 47217040 C13H20OSi 17.9 4-Trimethylsilyloxy-4-phenylbut-1-ene

35.06 86691232 C17H36O2Si 93.5 Tetradecanoic acid, trimethylsilyl ester

37.49 57155768 C18H38O2Si 68.5 n-Pentadecanoic acid, trimethylsilyl ester

39.19 138714112 C19H38O2Si 80 Palmitelaidic acid, trimethylsilyl ester

39.86 434806720 C19H40O2Si 94.2 Hexadecanoic acid, trimethylsilyl ester

43.59 113528232 C21H42O2Si 31 trans-9-Octadecenoic acid, trimethylsilyl ester

44.22 276898496 C21H44O2Si 94.7 Octadecanoic acid, trimethylsilyl ester

46.49 26053202 C23H50OSi 30.5 3,7,11,15-Tetramethyl-hexadecanol, trimethylsilyl ether

54.32 40401216 C27H56O4Si2 64.2 1-Monooleoylglycerol trimethylsilyl ether

55.48 16958486 C27H56O2Si 56.6 Tetracosanoic acid, trimethylsilyl ester

60.52 45148768 C30H54OSi 61.2 Cholesterol trimethylsilyl ether

64.69 51135540 C31H50P2 53.6 Methylenebis(2,4,6-triisopropylphenylphosphine)

(re p lib ) 3,6,9-Trio xa -2,10-d isila u n d e c a n e , 2,2,10,10-te tra m e th yl-50 80 110 140 170 200 230

0

50

10073

88

117

131147

160 191 207 235

OSi O

OSi

17.56

(m a in lib ) Be n ze n e , 1,1'-(1-m e th yle th ylid e n e )b is[4-(2-p ro p e n ylo xy)-50 90 130 170 210 250 290

0

50

100

55 77 91 115 165 183 211 252 277

293

308O

O 25.85

(re p lib ) n -Pe n ta d e c a n o ic a c id , trim e th ylsilyl e ste r50 90 130 170 210 250 290 330

0

50

100

55

73

98

117

129

201 224 255

299

314

O

OSi

37.49

(m a in lib ) Pa lm ite la id ic a c id , trim e th ylsilyl e ste r50 90 130 170 210 250 290 330

0

50

100

55

73

96

117129

145

194 236258 283

311

326

OSiO

39.19

(re p lib ) Te tra d e c a n o ic a c id , trim e th ylsilyl e ste r50 100 150 200 250 300 350

0

50

100

55

73

117

132

145

159 201 241

285

300

OO

Si

35.06

(m a in lib ) 4-Trim e th ylsilylo xy-4-p h e n ylb u t-1-e n e50 80 110 140 170 200 230 260

0

50

100

51

73

91 105 129 149

179

205

OSi

28.43

(m a in lib ) 2,4(1H ,3H)-Pyrim id in e d io n e , d ih yd ro -1,3-b is(trim e th ylsilyl)-50 80 110 140 170 200 230 260

0

50

100

59

73

93

100

114 131

147

171201

217

243

257

O N

N

Si

Si

O

22.58(re p lib ) Trim e th ylsilyl e th e r o f g lyc e ro l

50 90 130 170 210 250 2900

50

100

59

73

103

133

147

177

205

218293

OO

O

Si

SiSi

18.83

(m a in lib ) M e th yle n e b is(2,4,6-triiso p ro p ylp h e n ylp h o sp h in e )

50 110 170 230 290 350 4100

50

100

53 73 103 149 203 249279 355 395

441

PH PH

(re p lib ) C h o le ste ro l trim e th ylsilyl e th e r50 110 170 230 290 350 410 470

0

50

100

73

95

129

145

213 255

329

353

368458

OSi

(m a in lib ) Te tra c o sa n o ic a c id , trim e th ylsilyl e ste r50 120 190 260 330 400 470 540

0

50

100

55

73

97

117

132

201241 299 341 381

425

OO

Si

(re p lib ) 1-M o n o o le o ylg lyc e ro l trim e th ylsilyl e th e r50 120 190 260 330 400 470

0

50

100

55

73

103

129

147

203

257 307 339

397

485

OOO

Si

Si

O

(m a in lib ) 3,7,11,15-Te tra m e th yl-h e xa d e c a n o l, trim e th ylsilyl e th e r50 100 150 200 250 300 350

0

50

10057

83

125

139167 196 252

355OSi

(re p lib ) He xa d e c a n o ic a c id , trim e th ylsilyl e ste r50 90 130 170 210 250 290 330

0

50

100

55

73

83 97

117

132145

159 201 227 269

313

328

O

OSi

(m a in lib ) tra n s-9-O c ta d e c e n o ic a c id , trim e th ylsilyl e ste r50 100 150 200 250 300 350

0

50

100

55

73

84

117

129

145

185 222 264 295

339

354

O

OSi

(re p lib ) O c ta d e c a n o ic a c id , trim e th ylsilyl e ste r50 100 150 200 250 300 350

0

50

100

55

73

83

117

132

145

159 201 297

341

356

O

OSi

39.86

43.59

44.22

46.49

54.32

55.48

60.52

64.69

Aquarium Experiments• Tissue Dry Weight after the end of

the experimentSoft

Tissue Dry

Weight Student test Count Average p

Temperature

T1S1F2 Vs T2S1F2 5 824.4

223.3

80.6913

8

T1S2F2 Vs T2S2F2 8 923.0

324.5

7 0.6243

Salinity

T1S1F2 Vs T1S2F2 5 824.4

223.0

30.6847

1

T2S1F1 Vs T2S2F1 7 920.0

026.4

80.0892

8

T2S1F2 Vs T2S2F2 8 923.3

824.5

70.6598

6

FoodT2S1F2 Vs T2S1F1 8 7

23.38

20.00

0.24168

T2S2F2 Vs T2S2F1 9 924.5

726.4

80.5681

1

AgeAT1S2F2 Vs

T1S2F2 6 845.7

823.0

3 0.0001




Differencebefore

experiment

PALEOSALT 2008 Bivalves - VUB Rémy Mas, Fabrice Servaes, Maité Bauwens, Céline Poulain, Philippe...

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Transcript of PALEOSALT 2008 Bivalves - VUB Rémy Mas, Fabrice Servaes, Maité Bauwens, Céline Poulain, Philippe...