ars.els-cdn.com · Web viewIn August 2015, the ESB sampled 20 bream in the same stretch of the Danube that was also sampled in the present study. Sampling and processing were highly

Biota monitoring under the Water Framework Directive: On tissue choice and fish species selection

Annette Fliedner1, Heinz Rüdel1, Nina Lohmann2, Georgia Buchmeier3, Jan Koschorreck4

1 Fraunhofer Institute for Molecular Biology and Applied Ecology (Fraunhofer IME), Department

Environmental Specimen Bank and Elemental Analysis, 57392 Schmallenberg, Germany2 Eurofins GfA Lab Service GmbH, Neulaender Kamp 1a, 21079 Hamburg, Germany3 Bavarian Environment Agency (LfU), Demollstr. 31, 82407 Wielenbach, Germany4 German Environment Agency (Umweltbundesamt), 06813 Dessau-Rosslau, Germany

SUPPLEMENTARY MATERIAL

Part 1: Additional information

Part 2: Application of the conversion equation and -factors on comparative data

References

1

Part 1

Table S1: Biometrical characterization of fish sampled 2015 at Kelheim / Danube, and preparation of fish pools. Gray shaded fields: fish not included in pool.

a) chub

chub No.

length [cm]

weight whole fish[g]

Age [years]

sex weight carcass*

[g]

weighted portion carcass*

pool

fraction of pool

[%]

weight fillet [g]

weighted portion

fillet pool

fraction of pool

[%]

pool chub 115 40.0 843 5 F 642 87 17 192 92 1821 32.0 403 5 F 314 43 9 86 41 822 33.0 446 5 M 350 48 10 90 43 923 35.5 551 5 F 424 58 12 122 59 1224 33.0 470 5 M 369 50 10 93 44 925 32.5 361 5 M 275 37 7 81 39 826 38.0 683 5 F 537 73 15 139 66 1327 35.5 498 5 F 369 50 10 125 60 1228 35.0 515 5 M 394 54 11 117 56 11

pool chub 29 45.5 1350 7 F 1071 63 13 264 59 12

10 42.5 876 7 M 687 40 8 183 41 811 44.0 957 8 F 770 45 9 180 40 812 41.0 952 6 M 742 43 9 200 45 913 43.5 1143 6 M 885 52 10 248 56 1114 41.0 746 7 M 593 35 7 146 33 716 41.0 867 7 F 683 40 8 175 39 817 41.0 824 6 M 627 37 7 187 42 818 44.5 1216 7 F 955 56 11 246 55 1119 41.5 1009 7 F 791 46 9 199 45 920 42.0 961 6 F 754 44 9 192 43 9

pool chub 31 49.5 1923 11 F 1480 70 14 421 74 152 51.0 1878 9 F 1448 68 14 418 74 153 49.0 1745 9 F 1388 65 13 347 61 124 49.0 1657 10 F 1305 61 12 341 60 125 49.0 1649 10 F 1286 61 12 352 62 126 49.0 1841 9 F 1444 68 14 386 68 147 46.5 1463 9 F 1155 54 11 298 52 108 49.0 1403 9 F 1116 53 11 281 49 10

F: female; M: male, *included in carcass is one fillet

Table S1 (cont.): Biometrical characterization of fish sampled 2015 at Kelheim / Danube, and preparation of fish pools. Gray shaded fields: fish not included in pool.

2

b) bream

bream No.

length

[cm]


Age [years]

Sex weight carcass*

[g]


pool

fraction of pool

[%]

weight fillet [g]

weighted portion

fillet pool

fraction of pool

[%]

1 59.5 2840 12 F 2199 77 15 617 88 182 50.5 1333 9 F 1043 37 7 251 36 73 60.0 2696 12 F 2185 77 15 490 70 144 57.0 2615 10 F 2004 70 14 564 81 165 50.5 1542 11 F 1252 44 9 276 40 86 58.5 2449 10 F 1947 68 14 502 72 147 56.5 2451 11 F 1940 68 14 488 70 14

10 56.0 1993 9 F 1689 59 12 303 43 98 37 600.6 4 F 440.1 151.29 33 489.7 4 F 365 121.1

11 51.5 1670.4 8 M 1308.8 342.3

c) perch

perch No.

length [cm]


Age [years]

sex weight carcass*

[g]


pool

fraction of pool

[%]

weight fillet [g]

weighted portion

fillet pool

fraction of pool

[%]

pool perch 110 27.0 294 4 F 221 45 9 70 27 911 26.5 281 5 F 210 43 9 68 26 912 28.0 336 5 F 251 51 10 82 32 1113 27.0 333 4 M 257 53 11 73 29 1014 28.0 354 4 M 264 54 11 86 34 1115 27.5 321 4 F 241 49 10 76 30 1016 26.5 316 4 F 239 49 10 73 28 917 26.0 293 4 M 216 44 9 73 28 918 28.0 351 4 M 266 54 11 82 32 1119 29.0 368 4 F 278 57 11 87 34 11

pool perch 21 35.0 730 7 F 551 60 12 170 60 122 34.0 581 7 F 447 49 10 130 46 93 33.0 580 6 F 429 47 9 144 51 104 35.5 801 8 F 612 67 13 184 65 135 34.5 690 6 F 517 56 11 166 58 126 36.5 829 7 F 627 68 14 192 68 148 39.0 880 6 F 674 74 15 202 71 149 38.0 963 7 F 721 79 16 233 82 167 34.5 699 7 M 528.9 165.8

F: female; M: male, *included in carcass is one fillet

3

Table S2: Mercury concentrations [µg kg-1 wet weight] in chub (Squalius cephalus), bream (Abramis brama), and perch (Perca fluviatilis) sampled at Kelheim/Danube in 2015. Shown are measured concentrations in fillet and carcass* and calculated whole fish concentrations as well as fillet and whole concentrations normalized to 26% dry mass (DM).

fish species (number of

fish)

parameter Hg measured[µg kg-1 wet weight]

Hg normalized to 26% DM

[µg kg-1 wet weight]

fillet carcass* whole fish

WF-to-F ratio

fillet whole fish WF-to-F ratio

chub (n = 28)

MW 150 84.0 97.9 0.66 195 102 0.53SD 91.9 51.6 59.9 0.04 114 58.3 0.04

Median 125 66.1 77.6 0.67 170 86.5 0.54Min 44.8 24.7 29.0 0.55 57.7 31.8 0.43Max 349 212 242 0.72 431 232 0.59

bream (n = 11)

MW 174 91.3 107 0.63 233 109 0.48SD 113 58.3 67.1 0.05 171 77.3 0.03

Median 141 71.4 84.1 0.64 172 77.6 0.47Min 28.1 16.9 19.6 0.57 36.5 18.7 0.44Max 372 211 240 0.70 585 274 0.54

perch (n = 19)

MW 275 168 193 0.71 348 168 0.49SD 112 58.6 70.5 0.05 133 54.6 0.04

Median 258 170 192 0.71 341 171 0.49Min 131 80.9 93.1 0.64 173 92.2 0.42Max 509 301 348 0.86 605 275 0.59

*included in carcass is one fillet; WF-to-F ratio: whole fish–to-fillet ratio

4

Table S3: Significance of correlations between biometrical parameters and Hg concentrations in chub (Squalius cephalus, n = 28), bream (Abramis brama, n = 11), and perch (Perca fluviatilis, n = 19).

Colored fields mark comparable levels of significance: dark green: p < 0.001; bright green: p < 0.005; light olive: p < 0.05; light-green: p < 0.1; white: not significant

species parameter length weight whole fish

weight fillet weight carcass#

age sex Hg whole fish

Hg fillet Hg carcass#

WF-to-F ratio

DM fraction fillet

chub lengthweight whole fish <0.0001weight fillet <0.0001 <0.0001weight carcass# <0.0001 <0.0001 <0.0001age <0.0001 <0.0001 <0.0001 <0.0001sex 0.007 0.005 0.006 0.004 0.009Hg whole <0.0001 <0.0001 <0.0001 <0.0001 <0.0001 0.473Hg fillet <0.0001 <0.0001 <0.0001 <0.0001 <0.0001 0.401 <0.0001Hg carcass# <0.001 <0.0001 <0.0001 <0.0001 <0.0001 0.499* <0.0001 <0.0001Hg WF-to-F ratio 0.113* 0.116* 0.180* 0.100* 0.062* 0.150* 0.482* 0.244* 0.415DM fraction fillet 0.279 0.107 0.108 0.105 0.090 0.473* 0.128 0.068 0.163 0.005*DM fraction carcass# 0.004 0.001 0.001 0.001 0.001 0.247 0.003 0.001 0.004 0.022* <0.0001

bream lengthweight whole fish <0.0001weight fillet <0.001 <0.0001weight carcass# <0.0001 <0.0001 <0.0001age <0.0001 <0.001 0.001 <0.0001sex 0.486 0.403 0.429 0.396 0.353Hg whole 0.099 0.252 0.424 0.210 0.081 0.286*Hg fillet 0.094 0.253 0.438 0.208 0.095 0.254* <0.0001Hg carcass# 0.106 0.260 0.434 0.217 0.080 0.323* <0.0001 <0.0001Hg WF-to-F ratio 0.026* 0.114* 0.265* 0.087* 0.099* 0.139 0.027* 0.013* 0.036*DM fraction fillet 0.211 0.065 0.023 0.086 0.373 0.395 0.014* 0.016* 0.012* 0.253DM fraction carcass# 0.434 0.172 0.066 0.217 0.487 0.385 0.003* 0.002* 0.003* 0.069 <0.0001

*: negative correlation; DM: dry mass; WF-to-F ratio: whole fish-to-fillet ratio; #carcass includes one fillet

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Table S3 (cont.): Significance of correlations between biometrical parameters and Hg concentrations in chub (Squalius cephalus, n = 28), bream (Abramis brama, n = 11), and perch (Perca fluviatilis, n = 19).

Colored fields mark comparable levels of significance: dark green: p < 0.001; bright green: p < 0.005; light olive: p < 0.05; light-green: p < 0.1; white: not significant.

species parameter length weight whole fish

weight fillet weight carcass#

age sex Hg whole fish

Hg fillet Hg carcass#

WF-to-F ratio

DM fraction fillet

perch lengthweight whole fish <0.0001weight fillet <0.0001 <0.0001weight carcass# <0.0001 <0.0001 <0.0001age <0.0001 <0.0001 <0.0001 <0.0001sex 0.071 0.096 0.098 0.097 0.069Hg whole <0.001 0.001 0.002 0.001 0.015 0.376Hg fillet 0.001 0.002 0.002 0.002 0.025 0.379 <0.0001Hg carcass# <0.001 0.001 0.001 0.001 0.011 0.370 <0.0001 <0.0001Hg WF-to-F ratio 0.236* 0.310* 0.311* 0.311* 0.333 0.434 0.009* 0.002* 0.019*DM fraction fillet 0.053 0.066 0.068 0.065 0.070 0.053 0.030 0.026 0.034 0.081*DM fraction carcass# 0.002 0.005 0.007 0.005 0.016 0.153 <0.001 <0.001 <0.001 0.187* 0.009

*: negative correlation DM: dry mass; WF-to-F ratio: whole fish-to-fillet ratio; #carcass includes one fillet

6

Table S4: PFOS concentrations [µg kg-1 wet weight] in chub (Squalius cephalus), bream (Abramis brama), and perch (Perca fluviatilis) sampled at Kelheim/Danube in 2015. Shown are measured concentrations in pool samples of fillet and carcass* and calculated whole fish concentrations as well as fillet and whole concentrations normalized to 26% dry mass (DM).

pool Nfish in

pool

PFOS concentration [µg kg-1 wet weight]measured normalized to 26% DMfillet carcass* whole fish WF-to-F

ratiofillet whole fish WF-to-F ratio

chub 1 9 4.71 13.9 11.9 2.52 6.18 13.0 2.10chub 2 11 3.76 11.0 9.52 2.53 5.16 10.4 2.02chub 3 8 1.92 6.89 5.85 3.05 2.37 5.66 2.39bream 8 11.3 33.8 29.4 2.60 14.1 28.0 1.99perch 1 10 8.06 29.3 24.3 3.01 10.4 22.0 2.11perch 2 8 12.3 43.2 36.0 2.92 15.4 30.4 1.98

*included in carcass is one remaining fillet; WF-to-F ratio: whole fish–to-fillet ratio.

Table S5: Concentrations of lipophilic priority substances in chub (Squalius cephalus), bream (Abramis brama), and perch (Perca fluviatilis) sampled at Kelheim/Danube in 2015. Shown are measured and 5% lipid normalized concentrations in pool samples of fillet and carcass* and calculated whole fish concentrations. Number of fish in pool samples: chub 1: n = 9; chub 2: n = 11; chub 2: n = 8; bream: n = 8; perch 1: n = 10; perch 2: n = 8

substance pool concentrationsmeasured normalized to 5% lipid

fillet carcass* whole fish WF-to-F ratio

fillet whole fish

WF-to-F ratio

[ng kg-1 wet weight]PCDD/F + dl-PCB

chub 1 0.55 2.07 1.74 3.15 1.98 2.40 1.21chub 2 1.11 3.65 3.13 2.82 3.28 4.46 1.36chub 3 4.81 9.56 8.56 1.78 6.40 6.56 1.03bream 4.67 9.27 8.37 1.79 6.85 7.56 1.10perch 1 0.64 8.20 6.40 9.99 2.61 4.81 1.85perch 2 1.45 14.4 11.4 7.84 6.20 7.36 1.19

[µg kg-1 wet weight]ndl-PCB chub 1 8.62 25.4 21.7 2.52 31.01 30.03 0.97

chub 2 19.5 49.4 43.3 2.22 57.69 61.70 1.07chub 3 68.7 132 119 1.73 91.36 90.98 1.00bream 54.3 103 93.5 1.72 79.62 84.43 1.06perch 1 8.23 77.8 61.3 7.45 33.46 46.01 1.38perch 2 15.9 151 119 7.50 67.95 77.24 1.14

[µg kg-1 wet weight]PBDE chub 1 0.43 1.45 1.23 2.86 1.54 1.70 1.10

chub 2 0.77 2.39 2.06 2.68 2.27 2.93 1.29chub 3 2.95 7.56 6.59 2.24 3.92 5.05 1.29bream 4.13 9.38 8.36 2.02 6.06 7.55 1.25perch 1 0.66 8.28 6.47 9.83 2.67 4.86 1.82perch 2 1.36 15.3 12.0 8.85 5.81 7.79 1.34

[µg kg-1 wet weight]α-HBCDD chub 1 0.76 1.54 1.37 1.81 2.72 1.89 0.69


[µg kg-1 wet weight]HCB chub 1 0.31 1.20 1.01 3.24 1.12 1.39 1.25


*included in carcass is one remaining fillet; WF-to-F ratio: whole fish-to-fillet ratio.

7

Table S6: Significance of correlations between substance concentrations, stable isotopes, lipid, and dry mass in six pool samples of whole fish, fillet or carcass# (3 chub pools, 1 bream pool, 2 perch pools). Colored fields mark comparable levels of significance: dark green: p < 0.001; bright green: p < 0.005; light olive: p < 0.05; light-green: p < 0.1; white: not significant

whole fish

parameter δ13C δ15N lipid fraction

dry mass fraction

WHO(2005)-PCDD/F

WHO(2005)- dl-PCB

PCDD/F+dl-PCB

ndl-PCB PBDE α-HBCDD

HCB PFOS

δ15N 0.146*lipid fraction 0.393 0.063dry mass fraction 0.422* 0.018 0.001PCDD/F 0.406* 0.097 0.020 0.010dl-PCB 0.350 0.138 0.004 0.007 0.003PCDD/F+dl-PCB 0.381 0.135 0.005 0.007 0.002 <0.0001ndl-PCB 0.207 0.296 0.020 0.043 0.016 0.001 0.001PBDE 0.461* 0.066 0.006 0.002 <0.001 0.001 <0.001 0.012α-HBCDD 0.498* 0.386 0.290 0.235 0.048 0.152 0.129 0.153 0.118HCB 0.495* 0.061 0.002 0.003 0.014 0.012 0.012 0.039 0.011 0.177PFOS 0.083* 0.013 0.104 0.031 0.027 0.101 0.087 0.219 0.031 0.117 0.082Hg§ 0.369 0.063 0.003 0.003 0.030 0.005 0.007 0.022 0.007 0.366 0.025 0.102

#included in carcass is one remaining fillet

8

fillet parameter δ13C δ15N lipid fraction

dry mass fraction

WHO(2005)-PCDD/F

WHO(2005)-

dl-PCB

PCDD/F+dl-PCB

ndl-PCB

PBDE α-HBCDD

HCB PFOS

δ15N 0.146*lipid fraction 0.148 0.132*dry mass fraction 0.330 0.324 0.122PCDD/F 0.404 0.372* 0.012 0.059dl-PCB 0.160 0.224* <0.001 0.064 0.004PCDD/F+dl-PCB 0.186 0.240* 0.001 0.061 0.003 <0.0001ndl-PCB 0.107 0.170* <0.001 0.090 0.013 <0.0001 <0.001PBDE 0.362 0.351* 0.008 0.060 <0.0001 0.002 0.001 0.008α-HBCDD 0.353 0.242* 0.003 0.102 <0.001 0.002 0.001 0.007 <0.001HCB 0.143 0.176* <0.0001 0.083 0.014 <0.001 0.001 <0.001 0.009 0.005PFOS 0.062* 0.029 0.332* 0.240 0.299 0.449* 0.483* 0.355* 0.334 0.431 0.346*Hg§ 0.318 0.092 0.477 0.078 0.338 0.332 0.331 0.361 0.333 0.467 0.424 0.170

carcass# parameter δ13C δ15N lipid fraction

dry mass fraction

WHO(2005)-PCDD/F

WHO(2005) dl-PCB

PCDD/F+ dl-PCB

ndl-PCB

PBDE α-HBCDD

HCB PFOS

δ15N 0.146*lipid fraction 0.464 0.029dry mass fraction 0.395* 0.010 0.001PCDD/F 0.387* 0.063 0.020 0.008dl-PCB 0.396 0.089 0.005 0.005 0.003PCDD/F+dl-PCB 0.423 0.087 0.006 0.006 0.001 <0.0001ndl-PCB 0.248 0.216 0.025 0.035 0.012 0.001 0.001PBDE 0.445* 0.045 0.006 0.002 <0.001 <0.001 <0.001 0.008α-HBCDD 0.264* 0.360 0.348 0.266 0.067 0.183 0.158 0.176 0.143HCB 0.450* 0.033 0.002 0.002 0.013 0.011 0.012 0.039 0.011 0.210PFOS 0.087* 0.010 0.072 0.025 0.019 0.072 0.061 0.165 0.024 0.117 0.059Hg§ 0.384 0.056 0.002 0.003 0.017 0.001 0.002 0.009 0.003 0.347 0.018 0.086

*negative correlation; § refers to a hypothetical pool considering only those fish that were included in the pool samples; #carcass includes one fillet

9

Table S7: Influence of δ13C, δ15N and lipid whole fish-to-fillet ratio on the body distribution of substances in fish. Shown are significance levels (p-values) of correlations between the whole fish-to-fillet ratios based on measured concentration of PCDD/F+dl-PCB, ndl-PCB, PBDE, α-HBCDD, HCB, PFOS, and Hg, and δ13C, δ15N and the lipid whole fish-to-fillet ratio. Data refer to pool samples (3 chub pools, 1 bream pool, 2 perch pools) of fish sampled at Kelheim / Danube in 2015. Colored fields mark comparable levels of significance: bright green: p < 0.005; light olive: p < 0.05; light-green: p < 0.1; white: not significant

Whole fish-to-fillet ratios δ13C δ15N

Lipid whole fish-to-fillet

ratio

PCDD/F+dl-PCB 0.239* 0.007 0.007

ndl-PCB 0.257* 0.003 0.001

PBDE 0.270* 0.004 0.002

α-HBCDD 0.195* 0.002 0.002

HCB 0.252* 0.003 0.0002

PFOS 0.179 0.181 0.150

Hg§ 0.197* 0.012 0.011

Lipid 0.215* 0.0003*negative correlation; § refers to a hypothetical pool considering only those fish that were included in the pool samples

10

Table S8: Compliance of whole fish concentrations (wet weight ww) with biota quality standards. Data refer to fish sampled at Kelheim / Danube in 2015. PCDD/F+dl-PCB, ndl-PCB, PBDE, α-HBCDD, HCB, and PFOS data relate to pool samples of chub (Squalius cephalus, chub 1: n = 9, chub 2: n = 11, chub 3: n = 8), bream (Abramis brama, n = 8), and perch (Perca fluviatilis, perch 1: n = 10, perch 2: n = 8). Hg data refer to mean concentrations of individual fish (chub = 28, bream: n = 11, perch n = 19).

Substance sample Whole fish conc.

(measured)

Whole fish conc.

(normalized)*

EU EQS Environment Canada

(whole fish)

US wildlife valuese

(whole fish)PCDD/F+dl-PCB TEQ ng kg-1 ww

Chub 1 1.74 2.40 6.5Chub 2 3.13 4.46Chub 3 8.56 6.56Bream 8.37 7.56Perch 1 6.40 4.81Perch 2 11.4 7.36

ndl-PCB µg kg-1 ww

Chub 1 21.7 30.0 180 (otter)130 (mink)

440 (kingfisher)Chub 2 43.3 61.7Chub 3 119 91.0Bream 93.5 84.4Perch 1 61.3 46.0Perch 2 119 77.2

PBDE µg kg-1 ww

Chub 1 1.23 1.70 0.0085 32 (mammals)13 (birds)aChub 2 2.06 2.93

Chub 3 6.59 5.05Bream 8.36 7.55Perch 1 6.47 4.86Perch 2 12.0 7.79

α-HBCDD§ µg kg-1 ww

Chub 1 1.37 1.89 167 40000 (mammals)bChub 2 2.94 4.19


HCB µg kg-1 ww

Chub 1 1.01 1.39 10Chub 2 1.02 1.45Chub 3 2.36 1.81Bream 2.44 2.20Perch 1 2.82 2.12Perch 2 2.74 1.77

PFOS µg kg-1 ww

Chub 1 11.9 13.0 9.1 4.6 (mammals) 8.2 (birds)cChub 2 9.52 10.4


Hg µg kg-1 ww

Chub 97.9 102 20 33d 100 (otter)70 (mink)

30 (kingfisher)Bream 107 109Perch 193 168

*normalization to 5% lipid in the case of PCDD/F+dl-PCB, ndl-PCB, PBDE, α-HBCDD, HCB, respectively 26% dry mass in the case of PFOS and Hg §: biota quality standards refer to the sum of α-, β-, and γ-HBCDD of which about 80% is α-HBCDD in fisha: Federal Wildlife Dietary Guideline (FWiDG) for piscivorous mammalian and avian species (Environment Canada 2013). Refers to the commercial PentaBDE formulation DE-71b: Federal Wildlife Dietary Guideline (FWiDG) for piscivorous mammalian species (Environment Canada 2016)c: Federal Wildlife Dietary Guideline (FWiDG) for piscivorous mammalian and avian species (Environment Canada 2017)d: Canadian Tissue Residue Quality Guidelines for the Protection of Wildlife Consumer of Aquatic Biota (CCME 2017)e: U.S. Wildlife values (Lazorchak et al., 2003)

11

Fig. S1: Hg concentrations (µg kg-1 wet weight) in fillet of chub (upper row), bream (middle row) and perch (lower row) sampled in 2015 at Kelheim/Danube relative to fish age, length and weight. The frames highlight the areas with relative small variations in Hg concentrations.

Fig. S2: PBDE congener pattern in fillet and whole fish sampled at Kelheim / Danube in 2015. Data refer to pool samples of chub (Squalius cephalus, chub 1: n = 9, chub 2: n = 11, chub 3: n = 8), bream (Abramis brama, n = 8), and perch (Perca fluviatilis, perch 1: n = 10, perch 2: n = 8). F: Fillet, WF: whole fish

12

Fig. S3: Effects of normalization to 26% dry mass (DM) on concentrations of Hg (A) and PFOS (B) in fillet and whole fish of chub (Squalius cephalus), bream (Abramis brama), and perch (Perca fluviatilis) sampled 2015 at Kelheim / Danube. Hg refers to mean concentrations of individual fish (chub n = 28; bream: n = 11; perch: n = 19), PFOS refers to concentrations in pools samples (chub 1, n = 9; chub 2: n = 11; chub 3: n = 8; bream: n = 8; perch 1: n = 10; perch 2: n = 8). Dark columns: measured concentrations; light shaded columns: 26% DM normalized data.

13

Fig. S4: Concentrations in fillet vs. whole fish for pool samples of chub (Squalius cephalus, chub 1: n = 9; chub 2: n = 11, chub 3: n = 8), bream (Abramis brama, n = 8), and perch (Perca fluviatilis, perch 1: n = 10; perch 2: n = 8) sampled 2015 at Kelheim / Danube. Left side: measured concentrations of PCDD/F+dl-PCB (A), ndl-PCB (C), PBDE (E); HCB (G) and α-HBCDD (I). Right side: data normalized to 5% lipid (Fig. B, D, F, H, J). Concentrations refer to µg kg-1 wet weight, respectively ng kg-1 wet weight WHO(2005)-TEQ in the case of PCDD/F+dl-PCB. α-HBCDD (J): invalid regression

14

15

Part 2: Application of the conversion equation and –factors on comparative data

Two monitoring programs ran in parallel to the present study, one by the German

Environmental Specimen Bank (ESB), the other by the Bavarian Environment Agency (LfU).

In August 2015, the ESB sampled 20 bream in the same stretch of the Danube that was also

sampled in the present study. Sampling and processing were highly standardized following

the protocols of the ESB (www.umweltprobenbank.de). Bream were dissected and the fillets

were combined to one pool sample. Chemical analysis was consistent with the present

study.

The LfU sampled chub (n = 11) in September 2015 at a sampling site about 5 km upstream

of Kelheim (sampling site Kelheim oberhalb Klösterl, Danube River-km 2417 - 2420,

upstream of the confluence of Danube and Rhine-Main-Danube Canal). The samples were

analyzed by the LfU-laboratory. Hg was measured according to EPA method 7473 using a

dedicated atomic absorption spectrometer (DMA 80, Milestone). The sample was weighed

into a nickel vessel and oxidized by flowing oxygen in an electrically heated decomposition

furnace. The decomposition products are carried to an amalgamator, where mercury is

trapped and released into an atomic absorption spectrometer. Lipid determination was

performed gravimetrically according to the method described by Smedes (1999).

Ndl-PCB and HCB were determined by gas chromatography and mass spectrometry

(GC/MS) after pressurized liquid extraction (LfU 2013a). Liquid chromatography and mass

spectrometry (LC/MS/MS) was used for the analysis of PFOS (LfU 2013b). After pressurized

liquid extraction and a multi-column clean-up process, PCDD/F were analyzed by gas

chromatography and high resolution mass spectrometry (GC/HRMS) and dl-PCB and PBDE

by gas chromatography and tandem mass spectrometry (GC-MS/MS) (LfU 2016a). After

purification on silica/sulfuric acid followed by a Florisil column, HBCDD (sum of all isomers)

was determined in an aliquot of the same extract by GC-MS/MS with EI ionization (LfU

2016b). Chemical analysis was performed on a pool sample. The homogeneity of the pool

sample was controlled by additional measurements of Hg, ndl-PCB and HCB in the individual

fish.

The biometrical data of the fish and the respective data of the present study are summarized

in Table S9.

The ESB bream were slightly smaller and younger compared to bream of the present study,

their lipid content and TL, however, were slightly higher.

Chub sampled by the LfU were somewhere in between the chub in pools chub 1 and chub 2

of the present study with respect to size, age and fat content (Table S9). Their δ15N values,

however, were slightly higher indicating that they feed on a higher trophic level. 16

Table S10 summarizes the contaminant concentrations. Comparison of lipophilic substances

was based on 5% lipid normalized concentrations because only for these could conversion

equations and –factors be derived (Table 2, main paper).

Contaminant levels of ESB bream and bream in the present study were mostly within the

same range differing by no more than a factor of 1.8.

For LfU chub the homogeneity of the pool was verified (i.e., the calculated and measured

concentrations were in good accordance, Table S10).

Hg levels in fillets of the LfU chub were in good agreement with the data of pool chub 2 of the

present study. PFOS levels were slightly higher in LfU chub which may be related to the

higher trophic position of these fish (recall the TL-dependency of PFOS accumulation with

significant correlations p < 0.05 between δ15N and PFOS in all matrices, Table S6).

Concentrations of PCDD/F+dl-PCBs, PBDE and HCB in LfU chub were quite similar to the

concentrations in pool chub 1 while ndl-PCB levels were in between pools chub 1 and chub

2.

In the case of HBCDD, the LfU analyzed the sum of the HBCDD diastereomers whereas only

the α-diastereomer was determined in the present study. Nevertheless, the total HBCDD

concentration in LfU chub was clearly lower than the α-HBCDD levels in chub of the present

study. Differences in exposure but also in measurement may be responsible for these

discrepancies.

With respect to the calculation of the ESB- and LfU- whole fish concentrations, it must be

kept in mind, that the underlying data base of the conversion equations for lipophilic

substances and PFOS was relatively small and heterogeneous (n = 6, pool samples of three

species; different size classes). The resulting whole fish concentrations should therefore be

interpreted with caution. When applying the conversion equations for PCDD/F+dl-PCB, ndl-

PCB, PBDE and HCB on the fillet data of the present study the calculated whole fish

concentrations differed from the actually determined ones by up to ± 37 % (respectively up to

± 30% when using the conversion factors). For PFOS, however, the discrepancies were

smaller (max. ± 6%).

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Table S9: Biometrical data and characteristics of bream (Abramis brama) and chub (Squalius cephalus) sampled in 2015 at the Danube sites Kelheim (this study and the German environmental specimen bank ESB), respectively Kelheim oberhalb Klösterl (Bavarian Environmental Agency LfU).

Parameter Bream Chub 1 Chub 2 Chub 3 ESB bream LfU chubN fish 8 9 11 8 20 11Mean length [cm] 56.1 34.9 42.5 49.0 52.1 37.4Mean weight [g] 2240 530 991 1695 1676 667Mean age [years] 10.5 5.0 6.7 9.5 9.8 6.2Sex 8F 5F, 4M 6F, 5M 8F 10F, 10M 4F, 7MLipid (fillet) [%] 3.4 1.4 1.7 3.8 3.6 1.6b / 1.7c

dry mass (fillet) [%] 20.8 19.8 19.0 21.121,3b / 21,4c22b

/ c

δ13C -28.9 -28.6 -27.8 -26.2 -29.1 -29.6δ15N 14.5 13.6 13.3 13.0 14.7 14.1TLa 3.1 2.8 2.7 2.7 3.3 3.0

a: based on δ15N of zebra mussels sampled at Kelheim in 2014 b: refers to mean value of individual fish c: refers to pool

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Table S10: Concentrations of contaminants (wet weight) in fillets of bream (Abramis brama) and chub (Squalius cephalus) sampled in 2015 at the Danube sites Kelheim (this study and the German environmental specimen bank ESB), respectively Kelheim oberhalb Klösterl (Bavarian Environmental Agency LfU). Hg, PFOS: measured concentrations; all other substances: 5% lipid normalized concentrations. Whole fish concentrations for ESB bream and LfU chub were calculated using the conversion equations (CE), respectively the conversion factors (CF) given in Table 2.

contaminantmatrix Bream fillet /

poolChub fillet /

pool 1Chub fillet /

pool 2Chub fillet /

pool 3ESB bream fillet / pool

LfU chub fillet / individual fish

LfU chub fillet / pool

Hg µg kg-1

fillet 180.5a 85.6a 133.3a 252.2a 128.3 135b 120whole fish 116 56.3a 87.9a 160a 81.2 (CE 2) 94.5 (CF 0.7) 84 (CF: 0.7)

PFOS µg kg-1

fillet 11.3 4.71 3.76 1.92 12.9 8.5whole fish 29.4 11.9 9.52 5.85 36.3 (CE 6) 23.7 (CE 6)

PCDD/F+dl-PCB ng kg-1

fillet 6.85 1.98 3.28 6.40 7.65 1.97whole fish 7.56 2.40 4.46 6.56 9.94 (CF: 1.3) 2.6 (CF: 1.3)

ndl-PCB µg kg-1

fillet 79.6 31.0 57.7 91.4 65.5 41.3b 37.1whole fish 84.4 30.0 61.7 90.1 72.0 (CF: 1.1) 45.4 (CF: 1.1) 40.8 (CF: 1.1)

PBDE µg kg-1

fillet 6.06 1.54 2.27 3.92 4.83 1.41whole fish 7.55 1.70 2.93 5.05 6.76 (CF: 1.4) 1.98 (CF: 1.4)

α-HBCDD µg kg-1

fillet 5.73 2.72 2.70 3.75 10.2 0.85c (sum)whole fish 21.6 1.89 4.19 4.89 * *

HCB µg kg-1

fillet 1.36 1.12 0.95 1.46 0.82 1.21b 0.94whole fish 2.20 1.39 1.45 1.81 1.23 (CF: 1.5) 1.81 (CF: 1.5) 1.41 (CF: 1.5)

a:mean value of hypothetical pool that considers only those fish that were included in the pool sampleb : mean valuec : sum of α-, β-, γ-HBCDD* no valid conversion equation available

Conversion equations:Hg: conversion equation for bream (CE 2): ln (concwhole fish) = 0.9301 * ln (conc.fillet) – 0.1175

PFOS: conversion equation (CE 6): Conc.whole fish = 2.8459 * Conc.fillet - 0.4636

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References:

CCME Canadian Council of Ministers of Environment (2017): Canadian Environmental Quality Guidelines. Methylmercury. http://st-ts.ccme.ca/en/index.html?chems=133&chapters=all (accessed 4/2017)

Environment Canada (2013): Canadian Environmental Protection Act, 1999 Federal Environmental Quality Guidelines Polybrominated Diphenyl Ethers (PBDEs). http://www.ec.gc.ca/ese-ees/default.asp?lang=En&n=05DF7A37-1 (accessed 4/2017)

Environment Canada (2016): Canadian Environmental Protection Act, 1999 Federal Environmental Quality Guidelines Hexabromocyclododecane (HBCD). http://www.ec.gc.ca/ese-ees/default.asp?lang=En&n=8BA57E1C-1 (accessed 4/2017)

Environment Canada (2017): Canadian Environmental Protection Act, 1999 Federal Environmental Quality Guidelines Perfluorooctane Sulfonate (PFOS). http://www.ec.gc.ca/ese-ees/default.asp?lang=En&n=38E6993C-1 (accessed 4/2017)

Lazorchak JM, McCormick FH, Henry TR, Herlihy AT (2003): Contamination of fish in streams of the Mid-Atlantic region: An approach to regional indicator selection and wildlife assessment. Environ Toxicol Chem 22:545-553

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LfU (2013b): Medienübergreifende Umweltanalytik persistenter Perfluortenside und Bestimmung (semi)flüchtiger Vorläuferverbindungen. Report. Bayerisches Landesamt für Umwelt, Series UmweltSpezial. Pp. 92 (in German)

LfU (2016a): Etablierung eines GC-MS/MS-Gerätes mit erhöhter Massenauflösung für die Umweltanalytik und Anwendung bei Immissionsmessungen. Report. Bayerisches Landesamt für Umwelt, Series UmweltSpezial. Pp 31. (in German)

LfU (2016b): Untersuchungen zur Akkumulation verschiedener persistenter Schadstoffe in terrestrischen Wildtieren. Report. Bayerisches Landesamt für Umwelt, Series UmweltSpezial. Pp 66. (in German)

Smedes F (1999): Determination of total lipid using non-chlorinated solvents. Analyst 124: 1711-1718

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Documents

ars.els-cdn.com · Web viewIn August 2015, the ESB sampled 20 bream in the same stretch of the Danube that was also sampled in the present study. Sampling and processing were highly