Determination of heavy metals

International Journal of Environmental Science and Development, Vol. 2, No. 6, December 2011
Determination of heavy metals (Cd, Co, Cu, Ni and Pb) in
croacker fish (Johnius belangerii) from Musa estuary in
the Persian Gulf
Alireza safahieh, Fazel Abdolahpur Monikh, Mohammad Taghi Ronagh, Ahmad Savari, and
Abdolmajid Doraghi
28]. On the other hand, heavy metals accumulation by fish
may result in high level of metals intake by human consumers,
which finally leads to serious health problems.
Located in the northwest of the Persian Gulf, Musa estuary
is surrounded by port, harbor, and petrochemical complex as
well as urban areas. It is subjected to different types of human
activities such as petrochemical industries, ships and oil
tankers trafficking that are considered as potential sources of
heavy metals input into the seawater. On the other hand, this
estuary is an important location for fishing within the Iranian
coasts of the Persian Gulf.
Belanger's Croaker, Johnius belangerii is well distributed
in Musa estuary and its arbitraries and constitutes a
considerable portion of the annual catch of the local
fishermen. In addition, it is widely used by local people for
both direct and indirect consumption. Heavy metals
accumulation by fish with high annual catch may contribute
in sever health problems for its consumers. Since the
information concerning heavy metal accumulation in Musa
estuary is scarce, this study was carried out to determine the
concentration of Cd, Co, Cu, Ni and Pb in muscle, liver and
gill tissues of J. belangerii collected from different creeks of
the estuary. Besides, a comparison of our results with those
from other parts of the Persian Gulf is included to evaluate
the magnitude of the contamination in the area.
Abstract—Musa estuary is surrounded by port, harbor,
petrochemichal complex as well as urban areas and receives
various types of contaminants such as heavy metals. It is
considered as a habitat for Belonger’s croaker Johnius
belangerii in the Persian Gulf. Due to its relatively low price, the
fish is commonly consumed by the local people in Mahshahr
and Sarbandar cities, while there is no available data on heavy
metals accumulation by this species. This study was carried out
to determine the concentration of five heavy metals in J.
belangerii tissues. Fish samples were taken from five creeks and
dissected into muscle, liver and gills. The tissues were acid
digested and analyzed for their heavy metal contents. The
concentration of Cd, Co, Cu, Ni and Pb was determined by
flame atomic absorption spectrometer. Results showed that the
highest level of Cd, Co, Cu, Ni and Pb was 7.21, 1.08, 1.12, 2.72
µg/g and 4.57 in liver , 1.88, 1.04, 2.09, 9.43 and 6.83 µg/g in
gills and 0.14, ND, 5.61, 2.43 and 3.78 µg/g in muscle
respectively. The level of heavy metals in muscle was lower than
WHO standard, meanwhile the level of metals in fish were
decreased by the increase of distance from PETZON.
Biomonitoring of contaminants in Musa estuary could serve as
a good estimate of environmental health.
Index Terms—Musa estuary, havy metals, Johnius belangerii,
Persian Gulf
I. INTRODUCTION
Contamination of aquatic ecosystems with heavy metals
receives particular attention [1]. Essential and non-essential
heavy metals may appear as potentially harmful elements for
most aquatic and terrestrial organisms at some concentrations
of uptake. They are considered as serious contaminants
because of their persistence and remarkable tendency to be
concentrated in organisms tissues [2]. These elements are
introduced into aquatic environment through different
sources such as petrochemical wastewater, agricultural
runoff, and urban effluents as well as tankers traffic [1].
Environmental factors such as pH, salinity, DO and
temperature [6, 5] as well as ecological factors such as sex,
size and feed habit play significant roles in heavy metal
accumulation in aquatic organisms [11, 2]. Fish is regarded as
an appropriate bio-indicator for metals concentration in
aquatic ecosystems [4, 29, 30], because it could concentrate
large amount of some metals in different tissues [15, 26, 27,
II. MATERIALS AND METHODS
A. Study Area
The samples of fish were collected from five different creeks
along Musa estuary during August 2010. The sampling
stations were chosen in Khor-Ghazale, Khor-Ahmadi,
Khor-Jafari, Khor-Zangi and Khor-Ghanam (Fig. 1).
B. Sampling and Sample Preparation
Using trawl net, 20 fish samples from the same size (19-25
cm total length and 54-66 g total weight) were collected from
each creek. Fish were transferred to the laboratory using
icebox and kept frozen at -20 o C until analysis. Before
analysis, the samples were thawed in room temperature, their
tissues including muscle, liver and gills were obtained and
oven dried at 90 o C for 12 hour until constant weight was
obtained. One gram of muscle, 0.5 gram of liver and gills
were digested in concentrated nitric acid. The digestion
procedure stated at 40 o C for 1 h and continued in 140 o C for
the next 3 h. After digestion the samples were cooled in
laboratory temperature and the remaining digested solutions
were made up to certain volume with double distilled water
Manuscript received October 12, 2011, revised October 13, 2011.
A. Safahieh, F. Abdolahpur Monikh, M. T. Ronagh and A. Doraghi and
with the Department of Marine Biology, Faculty of Marine Sciences and
Oceanography, Khorramshahr University of Marine Science and
Technology, Khorramshahr, Iran (email: a.Safahieh@Kmsu.ac.ir.)
460
International Journal of Environmental Science and Development, Vol. 2, No. 6, December 2011
and finally filtered through 42µm filter paper.
To determine the metals concentration in the samples, a
GBC (Savant AA Sigma) flame atomic absorption
spectrometer (AAS) was used (Table I). All chemical regents
were from analytical reagent grade (Merck). All solutions
were prepared in deionized water. Calibration standards of
each heavy metal were prepared by appropriate dilution of
the stock solutions (1000 ppm, Merck). The glassware and
plastic containers were acid washed with nitric acid 10% for
24 h and rinsed with double distilled water before use. To
avoid samples contamination and check the accuracy of the
method, blank samples and CRM (Dorm-2, muscle of
Dogfish, National Research Council of Canada) were
analyzed at the same time and with the same manner of the
fish samples. The Blanks were also used to auto zero the
apparatus and it was routinely analyzed after 10 metal
analysis. The recovery values of all metals were satisfactorily
fallen between 90% to 114% (Table II).
C. Data Analysis
As said, all data were subjected to Shapiro-wilk test in
order to check normal distribution. Significant differences
between heavy metals concentration in various stations were
determined using One-Way analysis of variance (ANOVA)
followed by Duncan post hoc test (α=0.05).
Fig. 1. A map showing of study area
TABLE I: THE OPERATION CONDITION OF THE AAS
Metals
Cd
Co
Cu
Ni
Pb
Wavelengh (nm)
228.8
240.7
324.7
232
217
Silt Width (nm)
0.05
0.2
0.5
0.2
1
Lamp Current (MA)
3
6
3
4
5
TABLE II: HEAVY METALS LEVELS (µG/G D. W) OBSERVED IN DOGFISH
MUSCLE CERTIFIED REFERENCE MATERIAL DORM-2 (NATIONAL RESEARCH
COUNCIL, CANADA)
Dorm-2 (muscle)
Cd
Co
Cu
Ni
Pb
Observed values
0.048
0.201
2.67
17.12
0.083
Gas
Acetylene
Acetylene
Acetylene
Acetylene
Acetylene
Support
Air
Air
Air
Air
Air
Khor-Jafari. The average concentration of the metals in liver
is shown in Table IV. Significant differences were obtained
for the concentration of Cd, Cu and Pb in liver among
different stations. The maximum concentration of Cd (7.21
µg/g), Cu (12.32 µg/g) and Pb (4.57 µg/g) was found in the
liver of fish from Khor-Zangi, Khor-Jafari and Khor-Ghazale
respectively.
Table V shows the concentrations of heavy metals in gills
tissue. There were significant differences between
concentrations of Cd Ni and Pb in gills tissue of fish from
different creeks. The highest level of Cd (1.88 µg/g), Ni (9.43
µg/g) and Pb (6.83 µg/g) was recorded in fish from
Khor-Zangi, Khor-Ghazale and Khor-Zangi respectively.
The data indicated that each tissue has a specific capacity
for heavy metals accumulation (Fig. 2). The order of metal
accumulation in different tissues was as follows: gills > liver >
muscle for Cd, Ni and Pb, liver > gills > muscle for Co and
liver > muscle > gills for Cu.
Certified values
0.043
0.182
2.34
19.4
0.065
III. RESULTS
The average concentrations of studied heavy metals in
Johnius belangerii from Musa estuary (µg/g d.w) in muscle
are shown in Table III. The concentration of Co in muscle
was below the detection limit of the AAS. There were
significant differences between the concentration of Cu and
Pb in fish from different stations. The highest level of Cu
(5.61 µg/g) and Pb (3.78 µg/g) was measured in the fish from
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International Journal of Environmental Science and Development, Vol. 2, No. 6, December 2011
TABLE III: HEAVY METAL CONCENTRATION IN MUSCLE (µG/G DW)
Location
Khor-Ghazale
Khor-Ahmadi
Khor-Jafari
Khor-Zangi
Khor-Ghanam
a,b,c
Cd
0.09 ± 0.20
0.13 ± 0.03
0.08 ± 0.02
0.14 ± 0.03
0.10 ± 0.02
Co
Cu
Ni
a
ND
ND
ND
ND
ND
1.62 ± 0.07
3.59 ± 0.04b
5.61 ± 0.80c
1.68 ± 0.18a
1.53 ± 0.02a
2.04 ± 0.29
2.16 ± 0.55
2.43 ± 0.57
2.10 ± 0.52
1.19 ± 0.37
Pb
1.58 ± 0.20ab
2.42 ± 0.38bc
3.78 ± 0.76c
2.98 ± 0.61bc
0.59 ± 0.05a
Show differences among creeks, ND= Not Detected.
TABLE IV: HEAVY METAL CONCENTRATION IN LIVER (µG/G DW)
Location
Khor-Ghazale
Khor-Ahmadi
Khor-Jafari
Khor-Zangi
Khor-Ghanam
a,b,c
Cd
2.38 ± 0.42ab
1.09 ± 0.33a
1.8 ± 0.37a
7.21 ± 1.2c
4.23 ± 0.44b
Co
Cu
4.31 ± 0.32b
10.75 ± 1.21c
12.32 ± 1.76c
5.12 ± 0.45b
1.76 ± 0.42a
0.94 ± 0.12
1.08 ± 0.28
1.03 ± 0.21
0.98 ± 0.19
0.91 ± 0.12
Ni
2.39 ± 0.29
2.72 ± 0.33
2.53 ± 0.12
2.27 ± 0.14
2.53 ± 0.15
Pb
4.57 ± 0.35b
3.08 ± 0.41ab
3.97 ± 0.47b
3.31 ± 0.22ab
2.64 ± 0.38a
Show differences among creeks
TABLE V: HEAVY METAL CONCENTRATION IN GILL (µG/G DW)
Location
Khor-Ghazale
Khor-Ahmadi
Khor-Jafari
Khor-Zangi
Khor-Ghanam
a,b,c
Cd
Co
Cu
Ni
Pb
0.97 ± 0.04ab
0.92 ± 0.04ab
1.32 ± 0.11ab
1.88 ± 0.13b
0.73 ± 0.03a
0.91 ± 0.02
0.83 ± 0.03
1.04 ± 0.1
0.86 ± 0.02
0.8 ± 0.01
2.04 ± 0.25
1.94 ± 0.19
2.09 ± 0.43
1.37 ± 0.38
1.04 ± 0.26
9.43 ± 1.04c
7.89 ± 1.12b
6.74 ± 1.26b
1.63 ± 0.21a
6.82 ± 1.33b
5.71 ± 1.06b
5.33 ± 1.26b
6.83 ± 1.48c
6.62 ± 1.43c
2.57 ± 0.32a
Show differences among creeks
Fig. 2. Heavy metals concentration in different tissues of Johnius belangerii
(M: muscle, L:liver and G: gills)
IV. DISCUSSIONS
This study examined the concentrations of Cd, Co, Cu, Ni
and Pb in Muscle, liver and gills tissues of J. belangerii from
five creeks in Musa estuary. The heavy metal levels in J.
belangerii tissues showed different values among the stations.
The observed differences between metal concentrations in
the same tissues from different creeks indicate that there are
different sources for heavy metals input along Musa estuary.
In addition, physicochemical factors of the water from which
fish were caught may vary among studied creeks [9].
The relatively higher concentrations of some metals
recorded in Khor-Jafari could be related to the effluent
discharge from petrochemical units and Immam port [9]. A
big branch of Khor-Jafari has been dried by PETZON
(petrochemical special economic zone) and now serves as an
effluent receiving pond, where refined wastes are discharged
before releasing into the seawater. The high concentration of
Ni that was measured in Khor-Ghazale might be due to heavy
462
traffic of oil tankers in this creek.
Khor-Ahmadi are located between Khor-Jafari and
Khor-Ghazale, thus heavy metal in this creek may result from
dissolved metals in seawaters, which are transported via
water movement. Khor-Zangi also receives a huge amount of
petrochemical wastewater. In addition, this creek passes
Sarbandar and Mashahr cities before empting into Musa
estuary. Among the studied fish, those collected from
Khor-Ghanam were less contaminated. Since this creek is
relatively far from contamination sources in Musa estuary it
seems that low amount of contaminants could reach this area.
This study revealed that metal accumulation in gills and
liver occurs in higher magnitude than what appeared in the
muscle. This is a common finding that is also reported by
several investigations [13, 5, 1]. Because of the presence of
high levels of metallothionein protein, liver tissue acts as a
target organ for heavy metal detoxification [14, 8, 2, 1, 21].
Gills act as the main site for entry of different kinds of
contaminants such as heavy metals due to its continuous
contact with the external medium. This organ serve a variety
of physiological functions such as respiratory gas exchange,
osmoregulation and nitrogen excretion [19, 20].Therefore,
heavy metals may appear in a high level in liver and gill
tissues compared to what occurs in muscle. Muscle tissue has
lower tendency to accumulate heavy metals [22, 23, 24, 25].
Altindag and Yigit. (2005), Romeo et al. (1999) and Huang
(2003) found higher concentration of heavy metals in liver
and gill than in muscle.
A few data is available concerning heavy metals
concentration in fish species from north Persian Gulf
however, published data on J. belangerii are scarce. Doraghi
et al. (2009) reported concentrations of 0.62-0.78, 2.66-3.27,
9.48-11.78, 4.72-7.74, 1.82-3.22 µg/g dw for Cd, Co, Cu, Ni
International Journal of Environmental Science and Development, Vol. 2, No. 6, December 2011
[3]
and Pb respectively in J. belangerii collected from Hendyjan
and Deylam, in the northwest Persian Gulf [16]. The metal
concentrations in muscle of J. belangerii from Hendyjan and
Deylam were higher than those found in Musa estuary.
If compare the present results with metals concentration in
Epinephelus coioides studied in the south Persian Gulf [17],
it is clear that except Pb and Co in E. coioides from Bahrain
and Pb in the same species from Quatar the concentration of
other metals was noticeably higher in Cu and Ni measured in
the muscle of fish in present study was higher than what
reported for in J. belangerii . On the other hand metals
concentration in muscle samples were more or less the same
as the values reported for Euryglossa orientalis from the
same locations [18]. Rather than contaminant concentrations
in the ambient seawater, many other factors such as
contamination source, environmental factors, age, inherent
ability of the organism and even ecological niches may be
involve in metals accumulation by fish.
Although the maximum permissible level (MPL) has not
been established yet for all contaminants, heavy metals
concentration in edible tissue (muscle) of the studied fish (a
wet wt. dry wt ratio of 0.3) was below the MPL values of
Cu= 30 mg/kg and Pb= 2 mg/kg proposed by WHO [12].
Similarly heavy metals concentration in the muscle of J.
belangerii in the present study were much lower than the
MPL values of 0.5 mg/kg for Cd, 30 mg/kg for Cu and 2
mg/kg for Pb determined by FAO [12], indicating the
absence of serious health risk for consumers.
[4]
[5]
[6]
[7]
[8]
[9]
[10]
[11]
[12]
[13]
V. CONCLUSION
Present studies provide new information on the
concentration of heavy metals in commercial fish from Musa
estuary, the northwest Persian Gulf. Heavy metals
concentration in tissues of J. belangerii from Khor-Jafari
creek was higher than those fishes from other studied creeks.
This was presumably happened due to short distance between
the mentioned creek and contamination sources such as
petrochemical units and Immam port. Heavy metals
contamination in muscle tissue was lower than liver and gills.
The metals concentration found in the muscle (edible part of
fish) was lower than WHO and FAO standards for human
consumption. Biomonitoring of heavy metals in potentially
contaminated estuaries could serve as useful tools for the
awareness of environmental health.
[14]
[15]
[16]
[17]
[18]
[19]
ACKNOWLEDGMENT
[20]
This study was supported by Fajr Petrochemical Complex
of Mahshahr and Khorramshahr University of Marine
Science and Technology, Iran.
[21]
[22]
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