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Biomonitoring of heavy metals in fish from
the Danube River

Snjeana Zrni, Draen Orai, Marko

aleta, eljko Mihaljevi, Davor Zanella
& Nina Bilandi

Environmental Monitoring and

Assessment
An International Journal Devoted to
Progress in the Use of Monitoring Data
in Assessing Environmental Risks to
Man and the Environment

ISSN 0167-6369

Environ Monit Assess

DOI 10.1007/s10661-012-2625-x

1 23
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 Author's personal copy
Environ Monit Assess
DOI 10.1007/s10661-012-2625-x
Biomonitoring of heavy metals in fish from the Danube River
Snjeana Zrni & Draen Orai & Marko aleta &
eljko Mihaljevi & Davor Zanella & Nina Bilandi
Received: 24 August 2011 / Accepted: 2 April 2012
# Springer Science+Business Media B.V. 2012
Abstract The Croatian part of the Danube River
extends over 188 km and comprises 58 % of the
countrys overall area used for commercial freshwater
fishing. To date, the heavy metal contamination of fish
in the Croatian part of the Danube has not been stud-
ied. The main purpose of this study was to determine
heavy metal levels in muscle tissue of sampled fish
species and to analyze the measured values according
to feeding habits of particular groups. Lead ranged
from 0.015 g1 dry weight in planktivorous to
0.039 g1 dry weight in herbivorous fish, cadmium
from 0.013 g 1 dry weight in herbivorous to
0.018 g1 dry weight in piscivorous fish, mercury
from 0.191 g 1 dry weight in omnivorous to
0.441 g1 dry weight in planktivorous fish and arse-
nic from 0.018 g1 dry weight in planktivorous to
0.039 g1 dry weight in omnivorous fish. Among the
analyzed metals in muscle tissue of sampled fish, only
mercury exceeded the maximal level (0.5 mg kg1)
permitted according to the national and EU regulations
determining maximum levels for certain contaminants
S. Zrni (*) : D. Orai : . Mihaljevi : N. Bilandi
Croatian Veterinary Institute,
Savska 143,
Zagreb, Croatia
e-mail: zrncic@irb.hr
M. aleta : D. Zanella
Faculty of Science, Department of Zoology,
Rooseveltov trg 6,
Zagreb, Croatia
in foodstuffs, indicating a hazard for consumers of fish
from the Danube River.
Keywords Heavy metals . Danube River . Freshwater
fish . Mercury
Introduction
Globally, freshwater fish are the most heavily
exploited aquatic resources, representing about a quar-
ter (some 20 million tons per year) of the worlds food
from water (FAO 2008). Aquaculture production in
European countries is decreasing due to low produc-
tion efficiency. The decrease in aquaculture produc-
tion has inevitably led to an increase in fishing
pressures on freshwater fishery resources because of
increasing market demand for caught fish. A consid-
erable portion of the catch comes from recreational
fisheries, which increases the access of inhabitants to
fish from inland waters and leads to increased expo-
sure to all treats from insufficiently controlled open
waters.
Fish from open waters are considered wild ani-
mals as there is no possibility to control the compo-
sition of their growing environment (Clarkson 1998).
Those fish as part of the aquatic food chain are the
most likely route of human exposure to contaminants.
The frequency and intensity of water contamination by
heavy metals depends mainly on human activities and
their levels increase due to urbanization, industry,
 Author's personal copy
agriculture, or mining. Sometimes, contamination of
the aquatic environment can be of geological origin,
i.e., from ore-bearing rocks, forest fires, or vegetation
(Fernandez and Olalla 2000). Heavy metals may enter
the fish either through direct consumption of water or
organisms or by uptake through epithelia like the gills,
skin, and digestive tract (Burger et al. 2002). A third
possible route is via sediment (Bervoets et al. 2001)
which constitutes the most important reservoir of met-
als and other toxicants in aquatic environments. Fish
feeding on benthic organisms are directly exposed to
contaminated sediments while others are exposed
when toxicants from sediments are resuspended into
the water column. In the aquatic environment, metal
toxicity can be influenced by various abiotic factors
such as oxygen, hardness, pH, alkalinity, or water
temperature (Ghillebaert et al. 1995; Adhikari et al.
2006).
The Danube River, Europes second longest river
(2,857 km) flows through nine European countryes.
The Croatian part of the Danube is 188 km long and
forms part of Croatias eastern border with Serbia. The
Danube catchments area in Croatia comprises
32,800 km2 or 58 % of the countrys overall area and
it is widely used for commercial freshwater fishing. To
date, heavy metal contamination in the Croatian part
of the Danube has not been studied, while there are
some results of heavy metal research in the Danube on
fish caught in Germany, Austria, and Hungary (Wachs
2000): in mud, water, algae, and fish in the vicinity of
Vienna (Rehwoldt et al. 1975); in freshwater mussels
in the Vienna area (Gundacker 2000); in sediment in
the Hungarian part of the river (Gruiz et al. 1998); in
suspended solids and sediments along the entire river
flow (Woitke et al. 2003); in suspended solids, sedi-
ments, and mussels along the whole river (ICPDR
2002); in sediment and aquatic plants in the Serbian
part of the Danube (Pajevi et al. 2008); in liver, gills,
and skin of starlet at one sampling point in Hungary
and two in Serbia (Poleksic et al. 2010), and in the
same organs of Pontic shad from the Danube River in
Serbia (Vinji-Jefti et al. 2010). Studies have also
been conducted on the aquatic environment, aquatic
vegetation, macroinvertebrates, and fishes from the
Danube delta in Romania (Andreji and Stranai 2004;
Tudor et al. 2006; Diaconescu et al. 2008).
Traditionally, commercial and sport fisheries are
carried out in the Croatian part of the Danube River
and local inhabitants usually consume the caught fish.
Environ Monit Assess
Because of the highly developed industrial areas and
intensive agricultural production upstream and adja-
cent to the river flow in Croatia, the main goal of the
study was to determine heavy metal levels in sampled
specimens. The muscle tissues of several fish species
collected at three sampling points along the Danube
coast were analyzed with the aim of assessing the
possible health risk for humans. Another objective
was to compare the level of contamination in the
muscle tissue of sampled specimens depending on
feeding habits.
Material and methods
Sampling points
The study was performed at three points situated be-
tween where the Danube River enters Croatia at the
Hungarian border and where it leaves Croatia at the
Serbian border, in the period from April to October.
The first sampling point (Fig. 1) was on the border
with Hungary where the Danube River enters onto
Croatian territory, at the 1,436 river km (rkm) from
the river source (45 51 07.8N, 18 51 15.9E). The
river here is 420 m wide. The second sampling point
was situated 2 km downstream of the confluence of
the Drava River with the Danube River, near the
village Aljma at 1,478 rkm (45 3153.4N, 018
5716.3E). The river here is 650 m wide and the
influence of the Drava River on the Danube was also
investigated. Third sampling point was at 1,503 rkm
near Dalj (45 28 57.9N, 018 59 20.0E), where
the river is 540 m wide and near the point where the
Danube River leaves the Croatian territory.
Samples
All samples were collected only on the right bank,
approximately 2 m above the water surface by means
of electrofishing with a 10 kW generator. Samples for
physical and chemical parameters of the water at each
sampling point were determined at the time of collec-
tion of the fish. Determination of the species was
performed based on coloration patterns and other mor-
phological features using determination keys accord-
ing to Vukovi and Ivanovi (1971), Pov and Sket
(1990), and Maitland (2000). Total lengths of each fish
from the particular sample were measured to the
 Author's personal copy
Environ Monit Assess
Fig. 1 Fish sampling loca-
tions along the Croatian
part of Danube coast from
Hungarian to Serbian board
nearest millimeter with an ichthyometer (WILDCO
Fish Measuring Board, Ben Meadows, USA) and
weighed to the nearest 0.1 g using a digital scale
(OHAUS Scout pro, USA).
Fish for metal contamination analysis were stored
in the refrigerator at 4C on the spot and transferred in
cooling boxes to the laboratory where they were stored
at 18C until analysis. Fish were skinned, fileted, and
eviscerated. A commercial grade food grinder with
stainless steel cutting blades (7011HS, model
HGB2WTS3, Waring Commercial, USA) was used
to macerate samples. The ground tissue was then
homogeneously mixed before being sent through the
food grinder a second time. A subsample of the
ground tissue was then collected for analysis.
Reagents
All reagents were of analytical reagent grade, HNO3,
H2O2, and HCl (Analytical Grade, Kemika, Croatia).
Double deionised water (Milli-Q Millipore,
18.2 M cm1 resistivity) was used for all dilutions.
All plastic and glassware were cleaned by soaking in
diluted HNO3 (1/9, v/v) and were rinsed with distilled
water prior to use. Calibrations were prepared with
element standard solutions of 1,000 mg l1 of each
element supplied by Perkin Elmer. Stock solution was
diluted in HNO3 (0.2 %). As matrix modifiers in each
atomization for Pb and Cd, 0.005 mg Pd(NO3)2 and
0.003 mg Mg(NO3)2 (Perkin Elmer, USA) were used.
The hydride technique for mercury determinations
involves the reaction of acidified aqueous samples
(3 % v/v HCl) with a reducing agent 0.2 % sodium
borohydride in 0.05 % NaOH.
Determination of trace metal concentrations
Samples (2 g) were digested with 5 ml of HNO3 (65 %
v/v), 1 ml of H2O2 (30 %v/v) with a microwave closed
system Multiwave 3000 (Anton Paar, Germany). A
blank digest was carried out in the same way. The
program for the digestion began at a potency of
1,200 W then ramped for 10 min, after which samples
were held for 10 min at 1,200 W. The second step
began at a potency of 0 W and held for 15 min. Digested
samples were diluted to a final volume of 50 ml with
double deionised water. All metal concentrations were
determined on wet weight basis as milligram per
kilogram.
Analyses of As, Cd, and Pb were done by graphite
furnace atomic absorption spectroscopy using an Analyst
800 atomic absorption spectrometer (Perkin Elmer,
USA) equipped with an AS 800 auto sampler (Perkin
Elmer, USA). For graphite furnace measurements, argon
was used as the inert gas. Pyrolytic-coated graphite tubes
with a platform were used. The atomic absorption signal
 Author's personal copy
was measured in peak area mode against a calibration
curve. Mercury was analyzed by the cold vapor tech-
nique with a flow injection system coupled to a FIAS-
100 atomic absorption spectrophotometer (Perkin Elmer,
USA) equipped with an AS 93 plus auto sampler (Perkin
Elmer).
The limits of detection (milligram per kilogram) of
elements were found to be: As, 0.01; Cd, 0.0004; Cu,
0.0005; Hg, 0.0004; and Pb, 0.005. All specimens
were run in batches that included blanks, a standard
calibration curve, two spiked specimens, and one du-
plicate. In order to validate the method for accuracy
and precision, dogfish muscle (DORM-2, National
Research Council, Canada) was analyzed (n05) as a
certified reference material and the recovery (mean%
recoveryS.E.) was analyzed (n05). The recovery
was 93.36.3 % for As, 95.45.5 % for Cd, 98.7 pling points, which are situated very close to one anoth-
4.6 % for Cu, 98.13.6 % for Hg and 104.64.9 % for
Pb.
Statistical analysis
In order to assess differences in metal concentrations
in muscle tissue in different fish species according to
feeding habits, ANOVA with post hoc test analyses
based on Tukeys test was performed. When the
ANOVA assumptions were not met (data normality
and homoscedasticity), the non-parametric Kruskal
Wallis test was used. The relationship between fish
size (total length and weight) and metal concentration
in muscle tissue was investigated by univariate and
multivariate regression analysis. Statistical analyses
were performed using Stata 10.0 (StataCopr. 2005.
Stata Statistical Software: Release 10.0, College
Station, TX, USA). A level of p<0.05 was considered
significant.
Results and discussion
A total of 201 fish were chosen for heavy metal
concentration analysis from the three sampling points
in May, July, and September. Sampled specimens were
represented by 14 different fish species that varied in
total length from 83 to 860 mm and total weight from
6.3 to 7,000 g. The collected fish belonged to four
groups according to feeding habits (Table 1). The
water quality parameters, shown in Table 2, matched
the requirements for water quality category I
Environ Monit Assess
according to Croatian Water classification regulations
(Official Gazette 77/1998; Official Gazette 137/
2008b) excluding the measured value of O2 saturation
in a single sample at Batina and the concentration of
nitrates at the sampling site Dalj. Considering these
values, it is not possible to correlate the measured
metal concentrations with water properties as observed
in Bervoets et al. (2001), defined the influence of pH
on bioavailability of metals and increased bioaccumu-
lation of Hg. But it is widely known that temporal
oxygen deficiency enhances mobility of Hg in fresh-
water ecosystems (Boening 2000) and therefore it is
not possible to neglect the aberration of O2 saturation
even in a single measurement.
The results did not indicate any differences between
the measured metal concentrations from the three sam-
er. Statistical analysis showed that the differences
between heavy metal concentrations by sampling site
and time of sampling were not significant (p>0.05).
Metal concentrations and the corresponding mean
standard deviations (expressed as microgram per dry
weight) measured in muscle tissues are reported in
Table 1 for each fish species. Lead ranged from
0.015 g 1 dry weight in planktivorous to
0.039 g1 dry weight in herbivorous fish, cadmium
from 0.013 g 1 dry weight in herbivorous to
0.018 g1 dry weight in piscivorous fish, mercury
from 0.191 g 1 dry weight in omnivorous to
0.441 g1 dry weight in planktivorous fish and arse-
nic from 0.018 g1 dry weight in planktivorous to
0.039 g1 dry weight in omnivorous fish (Table 3).
Among the four metals analyzed in muscle tissue of
sampled fish, only mercury exceeded the maximum
level (0.5 mg kg1) permitted according to national
(Official Gazette 154/2008a) and EU regulations
(2006) determining the maximum levels for certain
contaminants in foodstuffs. This corresponds to the
results of Diaconescu et al. (2008) for analyzed muscle
tissues of mackerel and bream sampled at different
sites along the Danube River in Romania. The differ-
ences between the mean concentrations of measured
values in the four groups according to feeding habits
were significant (p<0.001). The highest concentra-
tions of mercury were expected in the muscle tissue
of piscivorous fish as described by Havelkova et al.
(2008), where predatory fish had much higher concen-
trations than non-predatory due to their position in the
food chain. In the present study, the highest mean
 Author's personal copy
Environ Monit Assess

Table 1 Number of specimen (n), their average total length (millimeter) and total weight (gram)standard deviation and range and
mean concentrationstandard deviation and range of lead, cadmium, mercury, and arsenic (microgram per dry weight) in muscle

Species according n Total Total Lead Cadmium Mercury Arsenic

the feeding habits length (mm) weight (g) MLa 0.30 (MLa 0.05) (MLa 0.50) (MLa 2.0)

Herbavour
Common nase 9 198.289.3 142.4272.7 0.0460.02 0.0080.005 0.1330.076 0.0220.098
Chondrostoma nasus 133411 20.3863 0.010.08 0.0050.018 0.0890.331 0.0110.045
Grass carp 9 251.676.8 282.6241.1 0.0340.022 0.0180.003 0.3670.231 0.0390.014
C. idella 126415 29.4886 0.010.08 0.0140.023 0.0910.937 0.0110.057
Omnivour
Ide 35 263.6104.7 343.0350.0 0.0470.029 0.0170.005 0.2130.008 0.0420.016
L. idus 83451 6.61,204 0.010.12 0.0120.029 0.0850.398 0.0110.069
Common bleak 10 1537.7 29.024.8 0.0250.022 0.0190.004 0.0970.01 0.0420.016
Alburnus alburnus 144, 172 23.638.2 0.010.08 0.0150.028 0.0870.118 0.0110.068
Roach 14 137.425.4 30.912.6 0.0280.019 0.0140.006 0.1520.028 0.0390.016
Rutilus rutilus 99162 13.643.2 0.010.06 0.0060.024 0.0890.185 0.0110.068
Prussian carp 11 23447.1 260.1169.7 0.0520.023 0.0170.004 0.0870.006 0.0310.011
Carassius gibelius 165327 90649 0.010.09 0.0110.024 0.0770.095 0.0110.045
Common carp 5 331211.2 1,316.62,343.9 0.0140.005 0.0160.004 0.2340.107 0.0550.054
C. carpio 118684 215,500 0.010.02 0.0110.022 0.0970.342 0.0210.15
Carp bream 37 200.362.7 121.5135.6 0.0300.02 0.0180.005 0.2370.121 0.0350.024
Abramis brama 108400 14.6742 0.010. 08 0.0070.029 0.0550.501 0.0110.15
Piscivour
Asp 32 267.3132.5 281.5417.2 0.0300.021 0.0200.004 0.2550.006 0.0330.016
Aspius aspius 93590 6.31,468 0.010.09 0.0140.029 0.080.79 0.0110.069
Pike 14 265.996.8 182.8190.3 0.0360.022 0.0150.005 0.2230.032 0.0300.015
Esox lucius 135444 17.4628 0.010.08 0.0070.022 0.1760.287 0.0110.065
Zander 12 191.838.2 60.135.3 0.0430.023 0.0180.002 0.1730.076 0.0320.014
Sander lucioperca 131256 16.2124 0.010.08 0.0150.023 0.0870.311 0.0170.057
Sheatfish 5 284.261.1 195.4138.2 0.0320.013 0.020.001 0.2350.128 0.0340.010
Silurus glanis 22038 90430 0.010.04 0.0190.022 0.1650.464 0.0210.048
Burbot 6 151.59.6 28.35.7 0.010.001 0.0200.002 0.1470.039 0.0480.015
Lota lota 142166 24.738.6 0.010.01 0.0180.021 0.1150.209 0.0270.068
Plankton-feeding fish
Silver carp 2 82056.6 6,0501,343.5 0.0150.007 0.0150.001 0.4410.007 0.0180.009
H. molitrix 780860 5,1007,000 0.010.02 0.0140.015 0.4360.446 0.0110.024
Burbot 6 151.59.6 28.35.7 0.010.001 0.0200.002 0.1470.039 0.0480.015
Lota lota 142166 24.738.6 0.010.01 0.0180.021 0.1150.209 0.0270.068
a
ML, maximal level permitted according to Comission Regulation 1881/2006 for Pb, Cd, Hg, and As in the muscles of fish intended for
human consumption

concentration of mercury was measured in the muscle
tissue of planktivorous fish, which could be explained
by the body size of the collected specimens, with body
lengths ranging from 780 to 860 mm and weighing
5,100 to 7,000 grams. These results are supported by
the findings of Jackson (1991) who indicated that
bioaccumulation of mercury is very size dependent.
The highest concentration of mercury in an individual
specimen was detected in the muscle tissue of two
specimens of asp (Aspius aspius; 0.716 and 0.79 g/
g dry weight) belonging to piscivorous fish, and one
specimen of grass carp (Ctenopharyngodon idella;
 Author's personal copy
Environ Monit Assess

Table 2 Water quality parame-

ters measured on the sampling Batina Aljma Dalj
points during the collection of fish Average, minmax Average, minmax Average, minmax
samples
Water temperature (C) 13.61 13.94 12.8
2.924.7 028 025
pH 8.07 8.04 8.22
7.68.4 7.558.6 7.968.67
El.conductivity (S/cm) 387.12 368.69 391.21
263454 237610 121487
Alkalinity (mg CaCO3/l) 155.12 153.65 149.33
105209 95207 115209
Dissolved O2 (mg/l) 11.94 9.96 9.09
7.315.8 6.214.6 6.911.9
O2 Saturation (%) 114.36 96.17 85.69
77.95a169.47 60159.61 58.69111.63
BOD5 4.11 3.84 2.95
1.28.7 0.78.7 0.65.9
Ammonium (mgN/l) 0.08 0.238 0.168
0.030.19 0.090.38 0.070.38
Nitrites (mg N/l) 0.0278 0.02 0.029
0.0140.05 0.010.03 0.010.32
Nitrates (mg N/l) 2.17 2.017 2.674b
0.93.62 0.6783.616 1.134.52
Total nitrogen (mg N/l) 2,783 2,584 3,157
1.594.69 1.24.18 1.515.05
o-phosphates (mg P/l) 0.0775 0.101 0.0722
a
Value characteristic for water 0.050.12 0.020.2 0.020.27
quality category II Total phosphorus (mg P/l) 0.143 0.187 0.247
b
Value characteristic for water 0.10.22 0.050.41 0.052.05
quality category III

0.937 g/g dry weight), belonging to herbivorous fish
(Table 1, Fig. 2). All sampled specimens were ana-
lyzed in respect to the time of sampling and the sam-
pling site (variables month and sampling point) in the
multivariate regression analysis. There was a signifi-
cant increase in mercury concentration with an in-
crease of fish total length (R00.0042, p<0.001) and
fish weight (R00.0000516, p<0.001), which corrobo-
rates previous findings, such as Lange et al. (1993),
who observed that fish age was more correlated with
mercury concentration than fish size, and Duek et al.
(2005), who found that age of analyzed individuals
and feeding strategy were the most important for Hg
accumulation in muscle tissue. It is widely known that
fish size is correlated with age. In this study, three
specimens weighed more than 5,000 grams, of which
two were silver carp (Hypophthalmychthys molitrix)
with a mercury concentration of 0.446 and 0.436 g/g
dry weight and one was common carp (Cyprinus car-
pio) with a mercury concentration of 0.342 g/g dry
weight (Table 1, Fig. 3). Those findings support the
thesis that it is not necessarily the case that predatory
fish are the best indicator species, as previously pro-
posed by Rincon-Leon et al. (1993). They studied the
importance of eating habits in the estimation of envi-
ronmental contamination through indicator species
and deduced that omnivorous species such as C. car-
pio, as a biological indicator of contamination, allows
for estimation with a greater confidence level than the
piscivorous species Anguilla anguilla. Similarly,
arski et al. (1995), and Svobodova et al. (1999)
concluded that benthophagous species feeding close
to the bottom are susceptible indicators of environ-
mental contamination depending to the degree of
 Author's personal copy
Environ Monit Assess

Table 3 Mean concentrationstandard deviation and range of lead, cadmium, mercury, and arsenic (microgram per dry weight) in
muscle of fish grouped according to their feeding habits

Feeding habit group n Lead Cadmiuma Mercury Arsenic

Herbavour 18 0.0390.022 0.0130.006 0.2500.206 0.0310.015

0.0340.046 0.0050.023 0.0890.937 0.0110.057
Omnivor 114 0.0360.026 0.0170.005 0.1910.101 0.0390.021
0.010.12 0.0060.029 0.0550.501 0.0110.15
Piscivor 69 0.0320.021 0.0180.004 0.2240.124 0.0340.015
0.010.09 0.0140.029 0.080.79 0.0110.069
Planktivor 2 0.0150.007 0.0150.001 0.4410.007 0.0180.009
0.010.02 0.0140.021 0.1150.446 0.0110.068
a
Differences between groups are significant (p<0.001)

sediment ingestion and direct exposure to contaminat-
ed particles.
A significant increase in lead concentration was
also observed with increasing fish total length (R0
0.0003 and p00.035); while for other metals (Cd and
As), no significant relationship with size was deter-
mined (p>0.05). The three highest lead concentrations
in muscle tissue were recorded in ide (Leuciscus idus);
0.1 and 0.12 g/g dry weight (Table 3, Fig. 4). Neither
of these measured values approaches the maximum
permitted level (ML) according to the EU Regulation
(1881/2006) and therefore its trends in different fish
species were not considered. The data presented here
differ from the results obtained by Jari et al. (2011),
who measured slightly increased cadmium levels in
the muscle of starlet (Acipenser ruthenus) from
Danube River in Serbia, while they did not analyze
mercury concentrations.
In fact, only mercury values were close to or
exceeded the ML of 0.5 mg Hg kg1, indicating a
hazard for consumers of fish caught in the Croatian
part of the Danube River. In 2003, the Joint FAO/
WHO Expert Committee on Food Additives adopted
a Provisional Weekly Intake level of 1.6 g/kg weight/
week (Smith and Sahyoun 2005), and therefore it
would be useful to create an exposure assessment
based on the quantity of fish caught either Official
Gazette in commercial fisheries or by angling.
Freshwater fish belong to the medium risk group in
terms of mean the mercury concentration (parts per
million) among commercial fish and shellfish (Hansen
and Dacher 1997). Various organ systems can be

Fig. 2 Concentrations of Hg
(microgram per gram dry
weight) in muscle tissue of all
sampled fish with respect to
fish total lenght
 Author's personal copy
Environ Monit Assess

Fig. 3 Concentrations of
Hg (microgram per gram
weight) in muscle tissue of
all sampled fish with respect
to fish weight

exposed to low doses of mercury causing damage to
the nervous, motoric, renal and cardiovascular, or im-
mune system (Zahir et al. 2005) comparison to some
marine fish species like tuna, mackerel, or swordfish.
Conclusions
& Among the four metals analyzed in the muscle tissue
of sampled fish, only mercury exceeded the maxi-
mum level permitted according to national and EU
regulations in asp, bream, and grass carp.
& Differences between the mean concentrations of
measured values in the four groups according to
feeding habits were significant.
& The highest concentration of mercury in an indi-
vidual specimen was detected in the muscle tissue
of asp (A. aspius) belonging to the piscivorous fish
and a specimen of grass carp (C. idella).
& There was a significant increase in mercury con-
centration with increasing total length and weight
of fish.
& Due to the increased values of mercury in muscle
tissue, it would be useful to establish an exposure
assessment based on the quantity of caught and

Fig. 4 Concentrations of Pb
(microgram per gram dry
weight) in muscle tissue of
all sampled fish with respect
to fish total lenght
 Author's personal copy
Environ Monit Assess
consumed fish either through commercial fisheries
or angling.
Acknowledgments This study was carried out as a part of the
project Management of freshwater fisheries on bordering rivers
pilot study with a holistic regional approach supported by the
Croatian Ministry of Agriculture, Forestry and Water Management
with the Norwegian partner, Akvaplan-niva AS.
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