Levels of non-dioxin-like PCBs in freshwater fish from the Danube

Full text

Environ Monit Assess
DOI 10.1007/s10661-010-1820-x
Non-dioxin-like PCBs in ten different fish species
from the Danube river in Serbia
Saša Jankovi´
c·Marijana ´
Curˇ
ci´
c·Tatjana Radiˇ
cevi´
c·
Sr ¯
dan Stefanovi´
c·Mirjana Lenhardt ·
Ksenija Durgo ·Biljana Antonijevi´
c
Received: 30 April 2010 / Accepted: 22 November 2010
© Springer Science+Business Media B.V. 2010
Abstract This work has been developed to exam-
ine the level of non-dioxin-like (ndl) PCBs (28,
52, 101, 138, 153 and 180) in (a) ten different
freshwater fish species from the Danube river, (b)
two sampling points: up and downstream of the
industrial zone of the city of Pancevo (ecological
hot spot in Serbia) and (c) two time points i.e.,
in 2001 and 2006. Obtained results would serve
to analyse spatial, temporal and congener profile
S. Jankovi´
c(
B)·T. Radiˇ
cevi´
c·S. Stefanovi´
c
Institute of Meat Hygiene and Technology,
Ka´
canskog 13, 11000 Belgrade, Serbia
e-mail: sasa@inmesbgd.com
M. ´
Curˇ
ci´
c·B. Antonijevi´
c
Department of Toxicology “Akademik Danilo
Soldatovi´
c”, Faculty of Pharmacy,
University of Belgrade, Vojvode Stepe 450,
11221 Belgrade, Serbia
M. Lenhardt
Institute for Multidisciplinary Research,
University of Belgrade, Kneza Višeslava 1a,
11030 Belgrade, Serbia
K. Durgo
Department for Biochemical Engineering,
Faculty of Food Technology and Biotechnology,
Zagreb University, Kršnjavoga 25,
1000 Zagreb, Croatia
characteristics of ndl PCBs cumulated in fish tis-
sues due to environmental pollution. Sixty-four
samples of the following species were collected:
wels (Silirus glanus), pike (Esox lucius), bream
(Abramis brama), crucian carp (Carassius caras-
sius), pike pearch (Stizostedion lucioperca), bar-
bel (Barbus barbus), tench (Tinca tinca), sterlet
(Acipenser ruthenus L.), common carp (Cyprinus
carpio) and bighead carp (Hypophthalmichthys
nobilis). Gas chromatography coupled with elec-
tron capture detector was used for analysis of
ndl PCBs. Total ndl PCBs content in upstream
samples ranged from 2.7 to 98.1 ng/g and from
4.9 to 68.3 ng/g in 2001 and 2006, respectively.
During the 5 years, ndl PCBs content increased
significantly in downstream samples i.e., ndl PCBs
varied from 13.7 to 46.1 ng/g and from 14.4
to 107.2 ng/g in 2001 and 2006, respectively.
PCBs 138 and 180 were predominant congeners
in 2001, while in 2006 the most abundant PCB
congeners were 138 and 153. In 2006, the pres-
ence of PCB 28 and PCB 52 has indicated a
recent contamination event. Data on continual
monitoring of PCBs in all relevant environmental
compartments together with appropriate biomon-
itoring data are expected to give comprehensive
insight into the fate and behaviour profile of these
contaminants.
Keywords ndl PCBs ·Fish ·Danube ·Serbia

Environ Monit Assess
Introduction
Polychlorinated biphenyls (PCBs) are a family
comprising 209 chemically related compounds
that were widely used in a variety of indus-
trial applications due to their insulating and fire-
retardant properties. Improper use and disposal,
as well as industrial accidents, due to the per-
sistence, liposolubility and tendency of PCBs to
bioaccumulate in animal tissues and milk resulted
in their spread in the environment often situated
remotely from the production or disposal site
(Smith and Gangolli 2002). Fish and other seafood
at all life stages, readily take up organochlori-
nated compounds, including PCBs, from water
with rather high bioconcentration factors making
them useful indicator organisms for the evaluation
of pollution of the aquatic environment. As a re-
sult of this bioconcentration, PCB levels in aquatic
organisms can be up to one million times higher
than their concentration in the aquatic environ-
ment. Accumulation of PCBs in fish depends on
their concentration in water, the life span, species
and fat content of the fish (Bayarri et al. 2001;
Bordajandi et al. 2006). Nowadays, they are of
great concern because of their persistence and
bioaccumulation through the food chain and their
toxic effects on wild life and humans. The main
human intake, almost 90% of PCBs, is via food
consumption (Liem et al. 2000; Lim et al. 2004).
Fish consumption, although with relatively small
share in total humans’ diet, compared with other
food products (meat and milk), can contribute up
to 50% of the intake of PCBs (Kiviranta et al.
2004; Antonijevic et al. 2007). Some social groups
may be exposed to higher PCBs concentration as
a result of their particular diet. Thus, the aquatic
environment is the major route of PCBs entry to
the food chain.
Different PCB congeners have different bi-
ological properties. Dioxin-like (dl) PCBs are
able to bind to the aryl hydrocarbon (Ah) re-
ceptor and have toxicological properties simi-
lar to those of the highly toxic compound 2,3,
7,8-tetrachlorodibenzo-p-dioxin. Non-dioxin-like
(ndl) PCBs have been shown to elicit different
types of responses than the dl PCBs, includ-
ing neurological, neuroendocrine, endocrine, im-
munological and carcinogenic effects (EFSA
2005). These effects occur via multiple toxicity
pathways, not involving the Ah receptor. There-
fore, unlike dl PCBs, no health based guidance
value for human exposure has been established
for ndl PCBs because their toxicity is still in-
sufficiently understood. Since ndl PCBs constitute
a major part of the PCBs found in food and
human tissues, several regulatory and advisory
boards have recommended that more information
should be gathered about the toxicity of ndl PCBs
(ATSDR 2000).
Some PCB congeners are highly persistent and
can, therefore, currently be found in the environ-
mental media, particularly in sediment and fish
matrices. Such congeners were considered as PCB
tracers, irrespectively of the dl or ndl PCBs types.
In this respect, analysis of food matrix contami-
nation profiles allows us to identify six most com-
monly found congeners (PCB-28, 52, 101, 138, 153
and 180), which account for approximately 50%
of all PCB congeners present in human food of
animal origin and in human fat, and are called
indicator PCBs (Arnich et al. 2009). Among these
indicator PCBs, the highly persistent di-ortho sub-
stituted PCB-138, 153 and 180 have been already
covered by specific regulations in some European
countries. The less persistent PCB-28, 52 and 101
were included since they were found in significant
amounts in some contaminated foodstuffs or were
considered as indicators of recent contamination
(Arnich et al. 2009).
United Nations Environment Programme
(UNEP)/United Nations Centre for Human
Settlement (UNCHS) established the Balkan
task force, with the aim of carrying out the
assessment of impact of the recent conflict on the
environment in Serbia (UNEP/UNCHS 1999).
The task-force, in the report, declared city of
Pancevo, (southern part of Serbian province of
Vojvodina, 15 km northeast from Belgrade-44◦52
N20
◦38E), as one of the ecological hot spots and
proposed immediate implementation of measures
to be taken in order to avoid serious consequences
on humans’ health. Additionally, Pancevo is a
city where a major industrial complex is situated
(petrochemical plant, fertilizer plant and major oil
refinery). These facilities are considered the most
important sources of pollutants in the Danube
river ecosystem. All these data, including data

Environ Monit Assess
on sediment concentrations of PCBs in this area,
ranging from 80 to 1,600 ng/g (UNEP/UNCHS
1999) have indicated that the levels of ndl PCBs
in fish could reflect environmental contamination.
Therefore, this work has been developed to
examine the level of ndl PCBs (28, 52, 101, 138,
153 and 180) in (a) ten different freshwater fish
species from the Danube river, (b) two sampling
points: up and downstream of the industrial zone
of the city of Pancevo and (c) two time points
i.e., in 2001 and 2006. Obtained results would
serve to analyse spatial, temporal and congener
profile characteristics of ndl PCBs cumulated in
fish tissues due to environmental pollution.
Experimental
During 2001 and 2006, 64 samples of ten different
fish species have been collected using random
sampling design: wels (Silirus glanus), pike (Esox
lucius), bream (Abramis brama), common carp
(Cyprinus carpio),cruciancarp(Carassius caras-
sius), pike pearch (Stizostedion lucioperca), bar-
bel (Barbus barbus), tench (Tinca tinca), sterlet
(Acipenser ruthenus L.) and bighead carp (Hy-
pophthalmichthys nobilis). Two locations were
chosen as sampling points: up and downstream
from city of Pancevo, Serbia (Fig. 1). According to
our knowledge, the upstream sampling site is free
of any major industrial complex that could cause
pollution in this location. The distance between
up and downstream fishing sites is approximately
300 km, allowing the minimization of the influence
of fish migration behaviour on results obtained in
this study.
After being caught, all fish specimens were kept
frozen at −20◦C before analysis. The head, tail,
backbone and fins were removed from the par-
tially frozen fish. Edible parts were chopped into
2 to 3 cm thick portions and homogenized.
Residues of PCBs in the fat extracted
from fish samples were analyzed according
to the USDA Analytical Chemistry Laboratory
Guidebook (1991). All standards and reagents
were purchased from Promochem (Wesel,
Germany). PCBs were extracted and separated
by elution from fat in small glass columns filled
with partially deactivated alumina. The eluate was
evaporated to an appropriate volume. An aliquot
Fig. 1 Sampling
locations, upstream and
downstream from
Pancevo, Serbia

Environ Monit Assess
Table 1 Freshwater fish
species collected from the
Danube upstream and
downstream from
Pancevo
Fish species and 2001 2006
number of samples Upstream Downstream Upstream Downstream
Carp +− ++
Pike +− +−
Barbel −− ++
Wels +− ++
Sterlet −+ ++
Bighead carp −− −+
Bream −+ −+
Pike pearch ++ −+
Tench −− −+
Crucian carp ++ −+
of 1 μL was injected into a gas chromatograph
coupled with electron capture detector.
Gas chromatograph GC Varian Model 3800
equipped with a 63Ni electron capture detector
(ECD) and Varian VF 5-ms column (30 m ×
0.25 mm i.d. and 0.25 μm film thickness) were
used for analysis of PCBs. Operating conditions
were as follows: injector 250◦C; detector 300◦C;
column oven program: initial 50◦C raised to 200◦C
at 50◦C/min, hold 2 min then raised to 215◦Cat
2.5◦C/min, hold 5 min and finally raised to 230◦C
at 2◦C/min, hold 9.5 min. The highly purified ni-
trogen carrier gas flow was 1 mL/min. Data acqui-
sition was performed by Varian Star software.
Table 2 Content of ndl
PCBs in the Danube fish
(ng/g fresh weight)
upstream and
downstream from
Pancevo (2001)
nd not detected
Species 28 52 101 138 153 180 PCBs
Upstream
Carp nd nd 1.7 10 5.9 18.1 35.7
Carp nd nd 3.8 14.4 9.4 24.8 52.4
Carp nd nd 2 6.6 4.6 8.8 22
Carp nd nd 1.4 8.4 4.5 9.2 23.5
Crucian carp nd nd 1.2 1.2 0.4 3.2 6
Crucian carp nd nd 4.4 11.2 6.7 51.2 73.5
Pike nd nd 1.4 2.1 1.5 2.6 7.6
Pike pearch nd nd 0.4 0.7 0.4 1.2 2.7
Pike pearch nd nd 0.5 1 0.5 1 3
Wels nd nd 4 16.5 9.7 23.7 53.9
Wels nd nd 1.2 6.2 3.8 3.7 14.9
Wels nd nd 8 27.9 16.6 45.6 98.1
Range – – 0.4–8 0.7–27.9 0.4–16.6 1–51.2 2.7–98.1
Mean – – 2.5 8.8 5.3 16.1 32.8
Median – – 1.6 7.5 4.6 9 22.8
Downstream
Bream nd nd 3 4.4 2.7 5 15.1
Bream nd nd 7.4 8.2 4.6 11.6 31.8
Bream nd nd 2.5 5.2 2.8 5 15.4
Bream nd nd 1.6 5 2.5 4.6 13.7
Crucian carp nd nd 6.7 3.3 2 2.1 14.1
Crucian carp nd nd 10.5 11.9 6.4 17.3 46.1
Pike pearch nd nd 2.2 2.8 1.6 14.1 20.7
Sterlet nd nd 4.6 6.2 3 4.4 18.4
Sterlet nd nd 4.4 9.9 5.3 9.8 29.4
Range – – 1.6–10.5 2.8–11.9 1.6–6.4 2.1-17.3 13.7–46.1
Mean – – 4.8 6.3 3.4 8.2 22.7
Median – – 4.4 5.2 2.8 5 18.2

Environ Monit Assess
Table 3 Content of ndl
PCBs in the Danube fish
(ng/g fresh weight)
upstream and
downstream from
Pancevo (2006)
Species 28 52 101 138 153 180 PCBs
Upstream
Barbel 2.6 3 10.7 23.9 19.4 8.7 68.3
Carp 0.6 1.1 2.5 7 8.8 4.5 24.5
Pike 0.5 0.5 1.5 0.5 1.4 0.5 4.9
Pike 0.9 0.9 2.4 11 12.2 5.1 32.5
Sterlet 1.7 1.4 8.1 5.8 6.8 2.2 26
Sterlet 1.6 1.4 8.1 5.6 6.6 2.5 25.8
Sterlet 0.8 1 6.8 4 4.1 1.1 17.8
Sterlet 1.2 1.6 6.4 5.8 5.5 2.1 22.6
Sterlet 1.8 1 6.4 5.2 4.5 1.7 20.6
Sterlet 1.6 1.3 5.6 5.9 5.9 3.1 23.4
Sterlet 1.3 1.3 6.5 5.2 4.6 2.3 21.2
Sterlet 1.7 1 6 3.8 3.9 1.2 17.6
Sterlet 1.8 1 7.6 4.8 4.2 1.7 21.1
Sterlet 1.6 1.3 7 9 8.4 3.2 31.5
Sterlet 2 1.5 8.2 7 7 5.5 31.2
Sterlet 2 1.2 8.3 10.8 10.6 7 39.9
Sterlet 1.6 1.3 6.6 7.1 6.6 3 26.2
Sterlet 1 1 5.1 6 5.9 3.1 22.1
Wels 1.3 1.3 4.5 21.2 17.6 9.8 55.7
Range 0.5–2.6 0.5–3 1.5–10.7 0.5–23.9 1.4–19.4 0.5–9.8 4.9–68.3
Mean 1.5 1.3 6.2 7.9 7.6 3.6 28
Median 1.6 1.3 6.5 5.9 6.6 3 24.5
Downstream
Barbel 1.3 1.9 6.2 9.6 9.9 4.4 33.3
Bighead carp 3.7 4.1 10.4 24.5 22.6 15.7 81
Bighead carp 3.5 4.8 7.4 15.6 14.4 6.3 51.8
Bream 2.3 2.6 6.6 10.7 11.6 6.6 40.4
Bream 2.2 2.4 15.9 7.2 6.1 3 36.8
Carp 5.2 4.4 10.7 40.9 34.3 11.7 107.2
Crucian carp 2.3 2 4.3 6.6 7.4 3.7 26.3
Crucian carp 2.3 1.5 2 6.4 6.5 3.6 22.3
Pike pearch 1.5 1.4 2.5 5.7 6.2 3.5 20.8
Pike pearch 2 2.2 6.9 9.4 8.7 2.8 32
Tench 1.8 2.5 2.1 2.6 3.7 1.7 14.4
Sterlet 6.5 8.1 13 14.1 14.1 10.2 52
Sterlet 6.4 12.3 18.4 21.5 22.4 13.3 94.3
Sterlet 2.9 6.1 8.5 11.5 11.5 9.5 50
Sterlet 3.9 5.9 7.8 10.8 10.4 6.6 45.4
Sterlet 3.3 5.4 8.9 11.8 11.9 7 48.3
Sterlet 1.1 1.7 7.5 12.8 12.5 3.8 39.4
Sterlet 1.4 1.8 5.4 8.9 9.1 5.2 31.8
Sterlet 2.1 2.4 8.1 9.3 9.1 6 37
Sterlet 5.8 9.1 7.9 12.3 11.6 9.7 56.4
Sterlet 3.5 3.9 3.4 7 7.2 5.6 30.6
Wels 2.8 3.7 11.9 13.7 14.4 10.7 57.2
Wels 2.3 3.1 9.6 35.7 37 12.5 100.2
Wels 2.5 3.1 12.2 28.5 23.6 8.6 78.5
Range 1.1–6.5 1.4–12.3 2–18.4 2.6–40.9 3.7–37 1.7–15.7 14.4–107.2
Mean 3 4 8.2 14 13.6 7.2 49.5
Median 2.4 3.1 7.8 11.2 11.6 6.4 42.9

Environ Monit Assess
Analysis of sample blank showed no interfer-
ence peaks with the individual PCB congener
analysis. The limit of determination for each con-
gener was determined as the mean of 10 times
background noise from five reagent blank sam-
ples. Methods limits of quantification, which
depended on congener type, were in the range
0.2–0.5 ng/g. Analytical quality control was
achieved by using certified reference material
ERM-BB446 (IRMM, Belgium). Accuracy and
intermediate precision were fulfilled according to
the specific requirements for determination of
ndl PCBs (Community Reference Laboratory for
Dioxins and PCBs in Feed and Food, Freiburg,
Germany 2008).
The concentrations of target individual con-
geners (IUPAC numbers 28, 52, 101, 138, 153
and 180) were expressed in ng/g of fresh weight.
Values below the limit of detection were assigned
to zero.
Descriptive statistics were estimated using
ORIGIN program (version 7.1). Mann-Whitney
U non-parametric test and principal component
analysis (PCA) were performed with Statistica
7.0 software. The differences were considered sta-
tistically significant when p value was less than
0.05.
Results and discussion
Non-dioxin-like PCBs levels in fish samples were
examined as a measure of freshwater pollution
nearby Pancevo. Additionally, the results ob-
tained from upstream and downstream samples
in two different time points could also give us
the data on both the influence of industry on
aquatic ecosystem and current/previous history of
pollution.
A summary of freshwater fish species used for
ndl PCBs measurement in this study is presented
in Table 1.
The content of ndl PCBs in edible tissues of fish
samples from the Danube is shown in Tables 2
and 3.
In 2001, the sum of the six indicator PCBs
in fish caught upstream from Pancevo was in
the range 2.7–98.1 ng/g, with a median value of
22.8 ng/g, whereas in 2006, PCBs content ranged
from 4.9 to 68.3 ng/g with a median value of
24.5 ng/g (Tables 2and 3). Medians do not differ
significantly, pointing out that the upstream levels
of ndl PCBs stayed unchanged during the five
60.26%
23.43%
8.81%
6.23%
0.94%
0.33%
012345678
0
1
2
3
4
5
PCB 28
PCB 52
PCB 101
PCB 138
PCB 153
PCB 180
-1,0 -0,5 0,0 0,5 1,0
PC1 : 60.26%
-1,0
-0,5
0,0
0,5
1,0
PC2 : 23.43%
Fig. 2 Eigen values correlation matrix and loadings plot of
PC1 (PCB 28) vs. PC2 (PCB 52) from the data set contain-
ing information on indicator PCBs (28, 52, 101, 138, 153
and 180) content in ten fish species matrices. *According
to Morrison (1967) principal components should account
for approximately 75% of the variables. Resulting loading
plot has indicated difference in distribution between c and
d patterns, what actually presents statistically higher values
of indicator PCBs concentration in downstream samples
in 2006 comparing with samples from not contaminated
location collected in the same time interval

Environ Monit Assess
years period. In contrast, ndl PCBs content in
fish caught downstream of Pancevo in 2006 was
significantly higher than in fish caught at the same
locations in 2001 (Table 2). Total ndl PCBs con-
tent in fish collected downstream of Pancevo in
2006 was in the range 14.4–107.2 ng/g, with me-
dian value of 42.9 ng/g, which was significantly
higher than in samples of fish caught upstream
during the same year (Table 3). Within the study
frame it was not possible to ensure statistically
relevant number of individuals of the same fish
species, allowing us to avoid variability due to
interspecies differences. Intraspecies comparison
was only possible to undertake in the case of
sterlets (Table 1). Results of statistical analysis of
the data on ndl PCBs concentrations in sterlets
were in accordance with the results obtained when
all species had been included, proving that in
2006 there was significant increase (p=0.0013)in
contamination in downstream samples related to
upstream ones.
PCA as the multivariate analytical tool is used
to reduce a set of original variables and to ex-
tract a small number of latent factors (princi-
pal components-PCs) for analyzing relationship
among the observed variables. Data submitted for
the analysis were arranged in matrices. For analy-
sis of correlation among locations of sampling
and collecting time 6 ×64 correlation matrix was
established. Furthermore, for analysis of correla-
tion between sampling locations in 2001, 6 ×12
and 6 ×9 matrices were established, respectively,
while for analysis of correlation between sampling
locations in 2006, 6 ×19 and 6 ×24 matrices were
established, respectively (Figs. 2and 3). Finally,
for analysis of correlation between the median
concentrations of ndl PCBs Mann–Whitney non-
parametric test was performed. Using PCA analy-
sis in certain combinations, results have shown
statistically significant difference between up and
downstream samples collected in 2006 that had
been already pointed out in first step of PCA
analysis (Figs. 2and 3).
For the purpose of comparison, levels of ndl
PCBs on freshwater fish published in available lit-
erature are presented in Table 4. Results similar to
ours were published by Vojinovic-Miloradov et al.
(2002) for pearch, carp and pike, also caught from
Fig. 3 Score plot of the
first two principal
component from data set
containing information on
indicator PCBs content in
ten different fish species
matrices. PC1 =PCB 28;
PC2 =PCB52 andl PCB
congeners in upstream
samples in 2001; bndl
PCB congeners in
downstream samples in
2001; cndl PCB
congeners in upstream
samples in 2006; dndl
PCB congeners in
upstream samples in 2006
a
a
a
a
a
a
a
a
a
a
a
a
b
b
b
b
b
b
b
b
bc
c
c
c
c
c
cc
c
c
c
cc
cc
c
c
c
c
d
d
d
d
d
d
d
d
dd
d
dd
d
d
d
d
d
d
d
d
d
d
d
-20246
PC1: 60.26%
-4
-3
-2
-1
0
1
2
PC2: 23.43%

Environ Monit Assess
Table 4 Literature data on the levels of ndl PCBs in freshwater fish
ndl PCBs Congener Fish species Source Reference
28 52 101 138 153 180
ng/g fresh weight (*ng/g lipids)
1,037.8 11.5 15.8 63 135 262.5 550 Roach Arrone, Italy Bazzanti (1997)
6–22.5 – – – – – – Perch, Carp, Pike Danube, Serbia Vojinovic-Miloradov et al. (2002)
370–1,100* – – – – – – Pearch Odra, Poland Falandysz et al. (2004)
8–177 1–5.6 3.3–5.2 0.9–0.65 0.4–4.5 1–1.8 0–2.4 different Sava, Croatia Bošnir et al. (2005)
7.8–56.9 – – – – – – different Drome, France Mazet et al. (2005)
7.48 0.14 0.3 0.94 2.2 2.4 1.5 Brown trout Redo, Spain Vives et al. (2005)
37–87 – – – – – – Pike pearch Nether land van Leeuwen et al. (2007)
1,451 7.6–233.1 0.5–124.5 3–159.7 14.8–509.8 10.3–226.7 10.3–200 Crucian carp China Zhao et al. (2007)
26.8 – – – – – – different France Arnich et al. (2009)
1.3–6.1 – – – – – – Pike pearch, Sulejowski Waszak and Dabrowska (2009)
Pearch reservoir, Poland
the Danube; Bošnir et al. (2005) for different fish
species from the Sava river which is the biggest
tributary of the Danube in Serbia; Mazet et al.
(2005) for fish from the Drome river (France);
van Leeuwen et al. (2007) for pike pearch from
the Lek river (Netherlands) and Arnich et al.
(2009) for freshwater fish from France. However,
in some other studies, PCBs concentrations were
either higher or lower than the levels found in
our study. For example, in the area for the dis-
assembly of obsolete transformers and electrical
waste in China, Zhao et al. (2007)aswellas
Bazzanti (1997) in river Arrone (Italy) after the
major contamination episode, determined 10–20
times higher levels of PCBs. In fish from freshwa-
ter reservoir in Poland (Waszak and Dabrowska
2009), or from high mountain lake Redo (Spain)
(Vives et al. 2005) reasonably lower concentra-
tions of PCBs were found due to low pollution
input.
Concerning the congeners’ profile, the pres-
ence of congeners 101, 138, 153 and 180 is evi-
dent in all the samples collected in 2001 (Fig. 4).
PCBs’ profile also showed that congeners 138
and 180 were predominant accounting for 27%
and 49%, respectively, in fish caught upstream,
and 28% and 36%, respectively, in fish caught
downstream of Pancevo. Congeners 28 and 52,
known as the indicators of recent exposure were
not detected in 2001, pointing out that there was
no additional contamination of the Danube river
by PCBs usually attributed to accidental release of
industrial sludge to the environment. In 2006, the
measurements indicated the presence of all indi-
cator congeners. Most abundant PCBs congener
was 138 accounting for 28% in both locations
(Fig. 4). The presence of PCB 28 and PCB 52
congeners has indicated a recent contamination
event. Congeners’ profile in our study are sim-
ilar to those published in several recent studies
(Bazzanti 1997;Zhaoetal.2007; Vives et al.
2005; Carubelli et al. 2007; Zuccato et al. 2008),
confirming the fact that highly persistent di-ortho
substituted PCB-138, 153 and 180 are the prevail-
ing congeners in biological samples.
During the 5-year period, there was an in-
crease in total ndl PCBs. Additionally, in 2006,
significantly higher concentrations were measured
in downstream samples, implying that the industry

Environ Monit Assess
Fig. 4 Congeners’ profile
for the six indicator ndl
PCBs
situated in the vicinity of Pancevo could be poten-
tial source of environmental contamination. Com-
plementary to this are the measurements of PCBs
in sediment revealing the concentrations from 80
to 1,600 ng/g (UNEP/UNCHS 1999). Obtained
results of our study can be of public concern since
there is a global trend of lowering the PCBs levels
in different media due to significant reduction of
their usage in industrial purposes (Bábek et al.
2008; Linderholm et al. 2010; El-Shaarawi et al.
2010). Besides, another issue related to the con-
sumption of fish and consequent human health
risk seems to be appropriate to address. Namely,
obtained ndl PCBs concentrations are below the
maximum residue level (MRL) of 3,000 ng/g
fresh weight, established by Serbian Anonymous
(1992), and are also below the limit of 2,000 ng/g
proposed by Food and Drug Administration,
USA (FDA 2001). For the sake of completeness,
it should be mentioned that no maximum levels
for ndl PCB in feed and food have been set in the
European Union so far. According to Commis-
sion Regulation (EC) No. 199/2006 (2006), MRLs
based on cumulative risk assessment and relative
toxicity regarding 2,3,7,8-TCDD, have been given
only for dl PCBs. The maximum level of 100 ng/g
fresh weight for the sum of 6 ndl PCBs in fish
has been proposed by the European Commission
(EC) draft regulation (AFSSA 2007). It can be
seen from the Tables 2and 3that, except two
samples collected downstream of Pancevo in 2006,
all the other values of ndl PCBs fall below the
European draft maximum level.
Conclusions
Results obtained in this work have shown tem-
poral increase in ndl PCBs levels in freshwater
fish caught from the Danube probably due to
certain environmental pollution. Even more, the
presence of PCB 28 and PCB 52 has indicated
a recent contamination event. Data on continual
monitoring of PCBs in all relevant environmental
compartments together with appropriate biomon-
itoring data are expected to give comprehensive
insight into the fate and behavior profile of these
contaminants. Further studies are also expected
to offer more precise assessment of biological
response i.e., inter- and intraspecies variability in
relation to bioaccumulaton capacity and congener
profile.
Acknowledgements This study is a part of the Project
No.TR20212A granted by Ministry of Science and Tech-
nological Development of the R. Serbia.

Environ Monit Assess
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