Metal Content in Blood of Free Ranging Harbor Seals

(Phoca vitulina) and their Effect on

Immunological Functions

Simone Griesel1; Antje Kakuschke1; Elizabeth Valenine-Thon2; Andreas Prange1

1 GKSS-Research Centre Geesthacht, Max-Planck-Straße 1, 21502 Geesthacht, Germany

2 Laboratory Dr. med. M.Sandkamp, B.Köster, Dr. med R.Hiller, MELISA (LTT) Center, Norderoog 2, 28259 Bremen, Germany

Fresh whole blood samples of 81 harbour seals (Phoca vitulina)

were collected between 2003 and 2005

Conclusions

- The concentrations of 27 elements in whole blood could be detected in the range of 1000 mg/L to 0,01 µg/L

- The levels of the essential elements P, S, K, Ca, Fe are reported in order to develop reference ranges for the health status

determination of live harbour seals

- The distribution patterns of trace elements reveal the actual body burden of seals in the German and Danish Wadden Seas

- The elements Al, Ti, Pd, Sn and Pt show significant geographical difference, but no significant age or sex related differences were found

- Seals with a Al-, Cr- and Ni- hypersensitivity also showed the highest concentrations of these metals in blood

Number of animals

location sex adult juvenile total number

Lorenzenplate m 20 12 32

f 5 17 22

L result 25 29 54

Römö m 21 1 22

f 3 2 5

R result 24 3 27

total number 49 32 81

age

Concentration of elements at Lorenzenplate

0,00

5,00

10,00

15,00

20,00

25,00

30,00

35,00

40,00

L 1 L 2 L 3 L 4 L 5 L 6 L 7 L 8 L 9 L 10 L 11 L 12 L 13 L 14 L 15 L 16 L 17 L 18 L 19 L 20

animal

concentr

ati

on ppb

Al Ti Pd Sn Pt

Concentration of elements at Römö

0,00

5,00

10,00

15,00

20,00

25,00

30,00

35,00

40,00

R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 R 13 R 14 R 15 R 16 R 17 R 18 R 19 R 20

animal

concentra

tio

n ppb

Al Ti Pd Sn Pt

0,01

0,10

1,00

10,00

100,00

1000,00

10000,00

100000,00

1000000,00

10000000,00

0 10 20 30 40 50 60 70 80

animal

Co

ncen

tratio

n o

f ele

men

ts (

log

pp

b)

P

S

K

Ca

Fe

Al

Ni

Pd

Cd

Sn

Pt

-Essential elements show less variation in element level

concentration (standard deviation = 12-25%) because of

homeostatic mechanisms

- Trace elements show wide variations (SD = 30-287%)

and therefore better reflect the exposure situation

Metal specific hypersensitivity

(black points)

coincided with elevated metal levels in blood

Geographical differencesFisher's Omnibus test 2 = 29,01 df = 16 P =0,023

Immunological Impact

Results

Total X-ray Fluorescence Spectrometry

(TXRF)P, S, K, Ca, Fe, Cu, Zn, As, Se, Rb, Sr

Inductively Coupled Plasma-Mass Spectrometry

(ICP-MS)collision cell

Be, Al, V, Cr, Mn, Co, Ni, Mo,

Pd, Cd, In, Sn, Pt, Pb, U

Ti

Memory lymphocyte immuno-stimulation assay

MELISA ®

Be, Al, Ti, Co, Cu, Mo, Pd, Cd,

Sn, Pt, EtHg, MeHg, HgCl, Pb

Lymphocyte transformation test (LTT) for

detecting antigen-specific metal sensitivities

Sector-Field ICP-MS (SF-ICP-MS)

Analytical Methods

Introduction

Sampling

- Marine mammals may serve as indicators of ecosystem change [Trilateral Dutch-German-Danish Monitoring and Assessment Program (TMAP)]

- Seals appear to be good indicators of accumulated hazardous substances in the marine environment

Element concentration

Chrom

0

10

20

30

40

50

60

Pv 2

25

6

Pv 2

25

7

Pv 2

25

8

Pv 2

26

4

Pv 2

47

4

Pv 2

47

5

Pv 2

47

6

Pv 2

50

8

Pv 2

50

9

Pv 2

51

0

Pv 2

51

2

Cr-

Ko

nze

ntr

atio

n (

g/L

)

0

1

2

3

4

5

SI

(Cr)

Nickel

< LOD0

5

10

15

20

25

Pv 2

256

Pv 2

257

Pv 2

258

Pv 2

264

Pv 2

474

Pv 2

475

Pv 2

476

Pv 2

508

Pv 2

509

Pv 2

510

Pv 2

512

Ni-K

onzentr

ation (

g/L

)

0

1

2

3

4

5

SI

(Ni)

µg/L Be Al P S K Ca Ti V Cr

Median 0.1704 14.89 439505 1534332 159877 42092 2.25 0.84 8.90

Min 0.092 0.169 153730 601494 96217 16060 0.58 0.32 0.358

Max 1.5 419.92 732078 2194144 199286 61920 10.80 4.48 115.52

Std % 111 199 24 21 14 22 78 59 132

µg/L Mn Fe Co Ni Cu Zn As Se Rb

Median 80.2 737819 0.74 3.84 809 3390 190 885 78

Min 41.18 471168 0.068 0.29 428 1914 46 459 55.9

Max 131.4 890749 5.04 33.58 1081 6264 780 2900 134

Std % 25 12 69 101 16 16 63 38 23

µg/L Sr Mo Pd Cd In Sn Pt Pb U

Median 46 5.066 0.840 0.212 0.100 0.440 0.210 1.82 0.081

Min 25.5 1.48 0.119 0.090 0.060 0.039 0.039 0.020 0.040

Max 95.00 21.62 13.80 0.600 0.294 4.00 2.45 8.10 0.360

Std % 30 47 113 50 69 102 127 64 70

With the collaboration of the FTZ Westcoast