Organohalogen concentrations and a gross and histologic ...
Organohalogen concentrations and a gross and histologic ...
Organohalogen concentrations and a gross and histologic ...
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Histology<br />
All tissue samples were trimmed, processed conventionally, embedded in<br />
paraffin, sectioned at about 4 µm <strong>and</strong> stained with Haematoxylin (Al-<br />
Haematein)-Eosin (HE) <strong>and</strong> periodic acid-Schiff (PAS) for routine diagnostics,<br />
Best's carmine to demonstrate glycogen storage <strong>and</strong> perls’ prussian blue<br />
reaction <strong>and</strong> Smorl for detecting haemosiderin <strong>and</strong> lipofuscin, respectively<br />
(Lyon et al., 1991, Bancroft <strong>and</strong> Stevens, 1996). All slides were examinated on<br />
a Leica DMLB microscope with 50, 100, 200, 400, 630 <strong>and</strong> 1000 x magnification<br />
<strong>and</strong> pictures were digitialised by a Leica DC300 digital camera.<br />
Histopathological changes (mononuclear cell infiltrations, granulomas, bile<br />
duct proliferation, portal fibrosis, lipid accumulation <strong>and</strong> necroses) were<br />
evaluated semi-quantitatively by examinating the liver tissue in 10 r<strong>and</strong>omly<br />
selected low to high power fields (200-400x magnification). Based on<br />
these observations each liver tissue was ascribed into one of three groups<br />
(absent-mild-moderate). Areas with severe freezing artifacts were omitted<br />
from evaluation.<br />
X-ray (osteodensitometry)<br />
X-ray osteodensitometry (DXA, Norl<strong>and</strong> XR 26) was applied to detect osteoporosis<br />
in the skulls of a sub-set of 44 polar bars. The method of determining<br />
bone mineral density (BMD) in these bears are described in detail in<br />
Sonne et al., (Accepted with revision).<br />
Analyses of PCBs <strong>and</strong> OCs<br />
Polar bear adipose tissue samples (n=67) were analysed for PCBs (Poly-<br />
ChlorinatedBiphenyls), DDTs (DichloroDiphenylTrichloroethanes), HCHs<br />
(HexaCycloHexanes), CHLs (CHLordanes), HCB (HexaChloroBenzene) <strong>and</strong><br />
dieldrin according to S<strong>and</strong>ala et al. (2004) <strong>and</strong> Dietz et al. (2004) at the Great<br />
Lakes Institute for Environmental Research (GLIER), University of Windsor,<br />
Canada. An external st<strong>and</strong>ard quantification approach used for PCBs <strong>and</strong><br />
OCs in the adipose tissues was based on peak area of the GC-µECD response,<br />
which is described in detail in Dietz et al. (2004). Briefly, ∑-PCBs is<br />
the sum (s) of the <strong>concentrations</strong> of the 51 individual or co-eluting congeners<br />
(if detected): CB # 31/28, 52, 49, 44, 42, 64/71, 74, 70, 66/95, 60, 101/84, 99,<br />
97, 87, 110, 151, 149, 118, 146, 153, 105, 141, 179, 138, 158, 129/178, 182/187,<br />
183, 128, 174, 177, 171/202/156, 200, 172, 180, 170/190, 201, 203/196, 195,<br />
194, 206. ∑DDTs is the sum of 4,4’-DDT, 4,4’-DDD <strong>and</strong> 4,4’-DDE. ∑-HCHs is<br />
the sum of the α-, β- <strong>and</strong> γ-hexachlorocyclohexane. ∑-CHLs is the sum of<br />
oxychlordane, trans-chlordane, cis-chlordane, trans-nonachlor, cis-nonachlor<br />
<strong>and</strong> heptachlor epoxide. Contaminant fractions were subsequently sent to<br />
the National Water Research Institute (Environment Canada, Burlington,<br />
Ontario, Canada L7R 4A6 (NWRI)) for determination of brominated diphenyl<br />
ether (PBDE) flame retardants.<br />
Analyses of PBDEs<br />
Polar bear adipose tissue samples (n=68) were analysed for BDPEs by electron<br />
capture negative ion (low resolution) MS using an external st<strong>and</strong>ard.<br />
Briefly, ∑-PBDEs is the sum (s) of the <strong>concentrations</strong> of the 35 individual or<br />
co-eluting congeners (if detected): BDE# 10, 7, 11, 8, 12/13, 15, 30, 32, 28/33,<br />
35, 37, 75, 71, 66, 47, 49, 77, 100, 119, 99, 116, 85, 155/126, 105, 154, 153, 140,<br />
145