Thiamine

1
Central archipelago
2

Biomarkers studies of female perch (Perca
fluviatilis) in a chronically polluted gradient
through the Stockholm archipelago L. Balk, B.
Liewenborg, M. Linderoth, H. Sundberg, E.
Noaksson, T. Hansson, U. Tjärnlund, G.
Åkerman. M. Hanson, Y. Zebühr Institute of
Applied Environmental Research, Stockholm
University, SE-106 91 Stockholm, Sweden D.
Schiedek et al. Baltic Sea Research Institute,
Seestrasse 15, D-18119 Rostock, Germany A.
Koehler et al. Alfred-Wegener-Institute for Polar
and Marine Research, Am Handelshafen 12, 27570
Bremerhaven, Germany J. Barsiene Institute of
Ecology, Vilnius University, Akademijos 2, 2600
Vilnius, Lithuania P. Vuorinen et al. Finnish
Game and Fisheries Research Institute, P.O.Box 6,
FIN-00721 Helsinki, Finland
3

--- Presentation outline ---
? Description of gradient
? Overview of biomarker data, previously
presented (recall Athens presentation)
? Further new biomarker data, not previously
presented
? Chemical analytical data
? -Conclusion-, -Future studies-
4

Description of gradient Stockholms aquatic
surrondings
BEEP-samples

• Downstream Stockholm
• Slussen 3 km
• 5) Kummelnäs 17 km
• 6) Torsbyfj. 26 km
• 7) Vindö 52 km
• 8) Lökholm. 72 km
• 9) Björksk. skärg. 84 km

Uppstream Stockholm 1) Adelsö 46 km
2) Färingsö 22 km 3) Riddarfj.
2 km
5

Map describing the 84 km gradient from downtown
Stockholm
Björksk. skg. (84)
Vindö (52)
Kummelnäs (17)
Lökholm. (72)
Torsbyfj. (26)
Slussen (3)
61 km
6

Normalization/standardization of sampling and of
biological material

• Fishing with
• gillnet
• mesh-size 35-40 mm
• 15 Sept.-15 Nov.
• ? Avoid migration
• ? Avoid interference
• with spawning
• ? Avoid minor obstacle such as ice
• ? Females is a major part of catch
• ? Well characterized
• time period for a
• number of biomarker

Female
Male
Female 19-39 cm
Female lt 19 cm
Female GSI 1 Sexually mature (SM)
Female GSI lt1 Sexually immature (SIM)
7

Overview of biomarker data, previously presented
Basic biodata/Supporting variables
/Physiological biomarkers/Biochemical variables
?
total length - no differences -
?
somatic condition factor - no differences -
?
spleen somatic index - no obvious differences
-
?
R20.95, Plt0,05
age - differences -
S
V
B
?
R20.83, Plt0,05
LSI - differences-
B
V
S
?
R20.99, Plt0,05
GSI - differences-
B
V
S
8

?
R20.90, Plt0,05
sexually mature adult females - differences-
V
B
S
R20.97, Plt0,05
S
?
visceral fat somatic index -differences-
B
V
K
R20.998, Plt0,05
?
blood haemoglobin - differences -
V
B
?
blood glucose - no differences -
R20.88, Plt0,063
R20.89, Plt0,22
?
CYP1A /EROD - differences -
B
S
V
DNA adducts - differences -
S
L
V
9

?
glutathione transferasee activity - no
differences -
?
glutathione reductase activity - no differences -
?
catalase activity - differences -
R20.89, Plt0,05
S
V
B
?
glucose-6-phosphate dehydrog. act. -difference-
S
K
?
CYP 19 activity- no differences -
?
acethylcholinesterase inhibition- differences -
R20.74, Plt0,06
V
S
10

Micronuclei in blood smears

Vindö
Slussen
Lökholmen
Kummelnäs
Björksk. skg.
Torsbyfjärden
Riddarfjärden
11

Lysosomal stability in liver tissue, peak 1
min.

Vindö
Slussen
Kummelnäs
Torsbyfjärden
(peak 2, same tendency although no significant
difference)
12

Apoptosis in liver tissue

Adelsö
Vindö
Slussen
Lökholmen
Färingsö
Kummelnäs
Björksk. skg.
Riddarfjärden
Torsbyfjärden
13

PAH conjugates in bile (1-OH-pyrene)
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Chemical analysis of a perfluorinated surfactant,
PFOS
K. Holmström, U. Järnberg, D. Berggren, C.
Johansson and L. Balk, Perfluorooctane sulfonate
concentrations i Swedish urban gradient fish
samples. Submitted manuscript.
15

Sum of 17 PCB congeners, equal amount (5-10gram)
epaxon muscle tissue pooled from 18 individuals
at each station
ng/g lipid
Sexually immature females
x 17
Adelsö
Vindö
Slussen
Lökholmen
Färingsö
Kummelnäs
Björksk. skg.
Riddarfjärden
Torsbyfjärden
34 km
84 km
Recalculated One percent lipid and 1/10 of liver
value gives 1mg/gram liver
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17 PCB congeners, what about human consumption?
Well, analogous to Aroclor 1254 and Food and Drug
Administration in Sweden the Recommended Daily
Intake is 20 ng/kg day
ng/g lipid
A kid of 20 kg1,2 gram
72 kg, 125 g portion, 1 lipid
Adelsö
Vindö
Slussen
Lökholmen
Färingsö
Kummelnäs
Björksk. skg.
Riddarfjärden
Torsbyfjärden
17

Sum of DDT, DDE, DDD, op-, pp- ng/g lipid, equal
amount (5-10gram) epaxon muscle tissue pooled
from 18 individuals at each station
ng/g lipid
Sexually immature females
x 6,8
Adelsö
Vindö
Slussen
Lökholmen
Färingsö
Kummelnäs
Björksk. skg.
Riddarfjärden
Torsbyfjärden
Comment not as steep gradients as for PCBs
18

Sum of 16 PBDE congeners, equal amount (5-10gram)
epaxon muscle tissue pooled from 18 individuals
at each station
ng/g lipid
Sexually immature females
na
Adelsö
Vindö
Slussen
Lökholmen
Färingsö
Kummelnäs
Björksk. skg.
Riddarfjärden
Torsbyfjärden
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Repeated chemical analysis for the year
2001 results in preliminary summary
? PCPs Slussen and Lökholmen, half levels
? PBDEs Slussen, Vindö and Lökholmen, identical
levels
? PFOS Slussen, Vindö and Lökholmen, identical
levels
20

Surface sediment exposure of Sea trout after ?2
days, showing the presence and bioavailable of
highly potent inducer(s) in central part of
Stockholms waterways presence
EROD activity
Vindö
Slussen
Control, no sediment
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Extraction and fractionation of anthropogenic
substances
Settling particulate matter
-Soxhlet extraction toluene-
Raw extract
-Silica-column-
Hexane fraction
Acetone fraction
-HPLC-amino column-
Monoaromat fraction
Diaromatic fraction
Polyaromatic fraction
22
Newly fertilized eggs are injected
Eggs are hold in 1 agarose gel
Endogenous carrier
23
Various disorders could be analyzed

24
Raw extract
CYP1A (EROD) directed fractionation Slussen.
97/98 seston
0.1 g
10 g
1.0 g
control
Acetone fraction
25
75
Hexane fraction
0.1 g
10 g
1.0 g
control
0.1 g
10 g
1.0 g
control
25
Hexane fraction (75)
Mono fraction
0.1 g
10 g
1.0 g
control
0
0.1 g
10 g
1.0 g
control
86
14
Di fraction
Poly fraction
0.1 g
10 g
1.0 g
0.1 g
10 g
1.0 g
control
control
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Conclusion lab study
The major toxicity could be followed from the
raw extract, via the hexane fraction, to the poly
fraction Lack of correlation to di fraction
strongly suggest dioxines, furans and PCBs not
responsible for the major potential adverse
biological effects in the area
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Conclusions
Surprisingly low biomarker response for some
biomarkers, while other shows a satisfactory
response through the entire 84 km long
gradient Relatively strong organ/tissue
response
show that biomarker works in pollution
gradients, if we now how to evaluate them
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Risk evaluation
High chronic exposure have caused resistance to
toxicity through the AhR receptor   Mechanism? U
ptake ? Immunology ? Genetic ? Metabolic
adjustment? Etc. ?   Long-term environmental
consequences ?   -UNKNOWN-
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STILL IN THERE ?! THANKS AGAIN ! !
Qs?
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Outer archipelago
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Percentage of adult female perch (19-39 cm) with
a developing gonad, i. e. sexually mature
females (n1000)
(?)
(?)

Adelsö
Vindö
Färingsö
Slussen
Klubben
Lökholmen
Riddarfjärden
Kummelnäs
Björksk. skg.
Torsbyfjärden
32

Upstream downtown Stockholm sites (km)
Färingsö (22)
Riddarfjärden (2)
Adelsö (46)
34 km
Downton Stockholm (Old town)