Determination and Comparison of Heavy Metals (Hg, Cd, Pb and As) of Barbusgrypus and Capoetacapoeta in Heleh River from Iran

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 Abstract—This Study was carried out to investigate
contamination of heavy metals Hg, Cd, Pb and As in muscle of
Barbusgrypus and Capoetacapoeta in Heleh river from Iran.
Heavy metal levels in fish samples were analyzed by Perkin
Elmer 4100 ZL atomic absorption. The highest concentration of
Hg, Cd, Pb and As were measured in muscle of Barbusgrypus
and the lowest concentration of Hg, Cd, Pb and As in muscle of
Capoetacapoeta. Concentrations of heavy metals Cd, Pb and As
in muscle of Barbusgrypus and Capoetacapoeta were showed
significant difference (P < 0.05), but for Hg there was no
significant difference (P > 0.05). Heavy metal concentrations
were higher in the Barbusgrypus, whereas compared with
Capoetacapoeta.
Index Terms—Barbusgrypus, Capoetacapoeta, heavy metal,
Heleh River, Persian Gulf.
I. INTRODUCTION
Fish are a major part of the human diet and it is therefore
not surprising that numerous studies have been carried out on
metal pollution in different species of edible fish [1]. In the
recent years, world consumption of fish has increased
simultaneously with the growing concern of their nutritional
and therapeutic benefits. The American Heart Association
recommended eating fish at least twice per weak in order to
reach the daily intake of omega- 3 fatty acids. Fish lipids have
also assumed great nutritional significance, because of their
high polyunsaturated fatty acid levels and good source of
digestible protein, vitamins and minerals. However, fish are
relatively situated at the top of the aquatic food chain;
therefore, they normally can accumulate heavy metals from
food, water and sediments [2].
Trace metals in aquatic environments may be of natural
origin from rocks and soil or from human activities [2], e.g.
industry [3], urban and agricultural discharge [4], mine runoff,
solid waste disposal and atmospheric deposition [5]. Heavy
metals (HMs) pollution of aquatic environment has become a
great concern in recent years. HMs can have toxic effects on
organs [6]. Heavy metals have the tendency to accumulate in
various organs of marine organisms, especially fish, which in
turn may enter into the human metabolism through
consumption causing serious health hazards. Iron, copper,
zinc and manganese are essential metals while, mercury, lead
and cadmium are toxic metals [7]. Heavy metals still play an
important role as pollutants affecting aquatic systems [8].
Metal enrichment of coastal waters mainly originates from
river inputs. Exposure to pollutants is therefore of prime
concern for biota inhabiting coastal areas, particularly during
their early development, which is very sensitive to toxic
effects of trace elements [9].
The content of toxic heavy metals in fish can counteract
their beneficial effects; several adverse effects of heavy
metals to human health have been known for long time. This
may include serious threats like renal failure, liver damage,
cardiovascular diseases and even death. Therefore many
international monitoring programs have been established in
order to assess the quality of fish for human consumption and
to monitor the health of the aquatic ecosystems [10].
Some of the metals found in the fish might be essential as
they play important role in biological system of the fish as
well as in human being, some of them may also be toxic as
might cause a serious damage in human health even in trace
amount at a certain limit. The common heavy metals that are
found in fish include copper, iron, copper, zinc and
manganese, mercury, lead and cadmium [9]-[11]. Toxic
elements can be very harmful even at low concentration when
ingested over a long time period. The essential metals can also
produce toxic effects when the metal intake is excessively
elevated [12].
In the last few decades, the concentrations of heavy metals
in fish have been extensively studied in different parts of the
world. Most of these studied concentrated mainly on the
heavy metals in the edible part. However, other studied
reported the distribution of metals in different organs like the
liver, kidneys, heart, gonads, bone, digestive tract and brain
[4].
This matter that, importance of the heavy metals measuring
relate to two important subjects which are aquatics ecosystem
management and human health, the present study was carried
out to determine the level of Hg, Cd, Pb and As in muscle
samples of Barbusgrypus and CapoetacapoetainHeleh River
from Iran. The fish and fish products for the people in those
ports are generally catch and carried by local vehicles from
the Heleh River. It should be noted that fish species are
considered to be a heavy metals part of the diet in the region.
The main objective of this study was to determinate the
contents of Hg, Cd, Pb and As in liver and muscle samples
Barbusgrypus and Capoetacapoetain Heleh River, in order to
assess fish quality and to assess the health risk for humans.
Determination and Comparison of Heavy Metals (Hg, Cd,
Pb and As) of Barbusgrypus and Capoetacapoeta in Heleh
River from Iran
Laleh Roomiani, Fatemeh Mashayekhi and Mansoreh Ghaeni
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Journal of Environmental Science and Development, Vol. 7, No. 9, September 2016
693
doi: 10.18178/ijesd.2016.7.9.864
Manuscript received June 4, 2015; revised October 27, 2015.
Laleh Roomiani and Mansoreh Ghaeni are with the Department of
Fisheries, Ahvaz Branch, Islamic Azad University, Ahvaz, Iran (e-mail:
L.roomiani@yahoo.com, Mansoreh.ghaeni@gmail.com).
Fatemeh Mashayekhi is with the Department of Food Science and
Technology, Khuzestan Science and Research Branch, Islamic Azad
University, Ahvaz, Iran (e-mail: Fatemeh_Mashayekhi68@yahoo.com).

II. MATERIAL AND METHODS
60 samples of Barbusgrypus and Capoetacapoeta were
collected from Heleh River of Bushehr Province. After
capture, fishes were placed in plastic bags with ice and
transported to the laboratory. Samples were cut into pieces
and labeled and then all sampling procedures were carried out
according to internationally recognized guidelines [13].The
procedure for the extraction heavy metals was based on
Standard Method 3052 (Microwave – assisted total heavy
metal digestion, USEPA, 1996). TORTE 2 (Lobster
Hepatopancreas Reference Material for Trace metals)
(National Research Council, Canada) was used as Standard
Reference Material for the digestion of fish, to verify the
accuracy of metal determination. The recovery rates were
90%± 10%. All of the digests were filtered through Whatman
5cc filter paper and stored at 4ºC. The concentration As, Cd
and Pb in sample solutions were determined by graphite
furnace atomic absorption spectrometry (GFAAS) and
hydride generation atomic absorption spectrometry
(HGAAS). The spectrometer (Analyst 800 coupled to FIAS
100, PerkinElmer, USA) uses transversally heated graphite
tubes and applies the Zeeman effect for background
correction.
The concentration of Hg was analysed by cold vapour
atomic absorption spectrometry (FIMS 100, PerkinElmer,
USA) at a wavelength of 253.7 nm. Generation of hydrides
for HGAAS (As, Cd) and reduction of Hg+2 to Hgo was
achieved by reaction of the sample solution with HCl (1 M)
and NaBH4 (6 gl-1As, Cd, 2gl-1Hg). Working standard
solutions for system calibration and control of analytical
accuracy were obtained by dilution of stock solutions (1000
mg/ l, Merck, Germany) using purified water (18 MΩ cm-1)
and analytical grade HNO3 (GFAAS) or HCl (HGAAS, cola
vapour AAS). Precision for determination of concentration
was better than 10% (Table I).
TABLE I: OPERATING CONDITIONS OF ATOMIC ABSORPTION SPECTROMETRY
W water, F fish, GFAAS graphite furnace atomic absorption spectrometry, HGAAS hydride generation atomic absorption spectrometry
a Temperature of trapping and drying for HGAAS
All reagents were of analytical reagent grade unless
otherwise stated. Double distilled water was used for the
preparation of solution. All the plastic and glass ware were
soaked in nitric acid for 15 min and rinsed with deionized
water before use. The stock solutions of metals (1000 mg/ L)
were obtained by dissolving appropriate salts of the
corresponding metals (E. merk) and further diluted prior to
use. High purity Argon was used as inert gas prior to use. The
samples were solubilized using high-pressure decomposition
vessels, commonly known as a digestion bomb. A sample (1g)
was placed in to Teflon container and 5 ml of concentrated
HNO3 was added. The system was heated to 130 ºC for 90 min
and finally diluted to 25 mL with deionized water. The sample
solution was clear. A blank digest was carried out in the same
way. Mercury, cadmium, lead and copper metals were
determined against aqueous standards.
T-test was run for all the collected data for fish samples
different using SPSS (17 version) computer programs. Mean
values of each parameter were compared using Fisher’s
protected least tests with significance levels of 5% were
conducted on each metal to test for significant differences
between sites (Table I). All statistical analyses were
conducted using the Office Excel 2003 software package.
III. RESULTS AND DISCUSSION
Concentration levels of metals Hg, Cd, Pb and As in muscle
of Barbusgrypus and Capoetacapoetawere measured and
presented in Table II. Concentrations of metals are presented
in mg/ kg dry weight unless otherwise mentioned. The highest
concentrations of Hg, Cd, Pb and As in tissues were measured
in muscle of B. grypus and the lowest concentration of Hg, Cd,
Pb and As were showed in muscle of C. capoeta (Fig. 1).
Concentrations of heavy metals Cd, Pb and As in muscle of
Barbusgrypus and Capoetacapoeta showed significant
difference (P < 0.05) but concentration of Hg not showed
significant difference (P > 0.05). Heavy metal concentrations
were higher in the Barbusgrypus, when compared with
Capoetacapoeta.
According to the priority List of Hazardous Substance
established by the Agency for Toxic Substances and Disease
Registry (ATSDR, 2013) [13], the descending order of heavy
metals threatening to human health were As>Pb> Cd> Ni>
Zn> Cr> Cu>Mn.
A variable range of different metal concentrations has been
observed by various researchers worldwide. The absorption
of metals on to the gill surface, as the first target for pollutants
in water, could also be an important influence in the total
metal levels of the liver [14], [15]. Distribution patterns of
metal concentrations in muscle of Barbusgrypus and
Capoetacapoeta from Heleh river follows the sequence: Pb>
As> Cd> Hg. There are various studies on the heavy metal
levels in fish from different waters. Minimal accumulation
and storage of heavy metals in such families on fish (Liza
auratus) in the southern Atlantic coast of Spain [16], on fish
(Mugilauratus) in the Black Sea Turkey [17], on fish (Liza
abu) in Lake Ataturk [18], on fish (Liza carinata) [19] is
proven. The observed variability of heavy metal levels in
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694

different species depends on feeding habits [20], ecological
needs, metabolism [21], age, size and length of the fish [22]
and their habitats [7]-[23]. Reference [24] studied the heavy
metal levels in kidney, liver, gill and muscle of Torgrypus and
Reference [25] studied the concentration of Zn and Pb in liver
of Cyprinuscarpio, C. aculeate and C. damasciana which
concentration of Zn were higher than Pb.
Reference [24] studied the heavy metal levels in muscle,
liver, gonad, and gill of gilthead seabream (Sparusaurata),
European sea-bass (Dicentrarchuslabrax), and keeled mullet
(Liza carinata) which concentration of Zn were higher than
Pb. The levels of Zn in all tissues were higher than the Pb
levels, as Zn is present in many enzymes throughout the fish’s
body.
In this study, concentrations of Hg, Cd, Pb and As in
muscle of Barbusgrypus higher than Capoetacapoeta.
Muscle tissue is the main edible fish part and can directly
influence human health. Lead enters into the body with gill
cells and especially is accumulated in gills and the later aim
organs are liver and muscle [26]. In other studies,
concentrations of Hg, As, Cd and Pb in liver of Liza
dussumieri higher than muscle [27].Level of mercury in liver
of Liza parsia was higher than muscle [28] also concentration
of heavy metals (Hg, Cd, Pb, Cu, Zn, Fe and Mn) in liver of
Liza abu was higher than muscle [29]. In other study such as
Mugilauratus [17], Sparusauratus, Triglacuculus,
Sardinapilchardus, Mugilcephalus, Atherinahepsetus,
Scomberesoxsaurus [7], Serranusscriba, Epinephelus costae,
Cephalopholisnigri and Pseudupenaeusprayensis [20],
Scomberomorusguttatus, Scomberomoruscommerson and
Otolithesruber [30], Barbusxanthopterus [31], Barbusgrypus
and Barbusxanthopterus [31] concentrations of heavy metals
in liver was higher than muscle.
Fig. 1. Comparison of heavy metals in muscle of Barbusgrypus and
Capoetacapoeta in Heleh river (mg/ kg) (a: non-significant differences at P
< 0.05, b: significant differences at P < 0.05).
The other research, concentrations of heavy metals (mean±
SD) Hg, As, Cd and Pb in muscle and liver of Liza dussumieri
from Boshehr seaport were higher than in Liza dussumieri
from Deylam seaport (P < 0.05), except for concentration of
Pb that in muscle of Liza dussumieri from Deylam seaport
were higher than in muscle of Liza dussumieri from Boshehr
seaport (P < 0.05) [27].
Reference [32] reported that in Barbusxanthopterus and
Barbusrajanorummystaceus in Ataturk Dam Lake, Turkey,
heavy metal concentrations in gill and liver were high while
these concentrations were the least in muscle. Reference [24]
has reported the concentrations of Co, Cr, Cu, Fe, Mn, Ni, Pb
and Zn in liver and muscle of Tor grypus in Ataturk Dam Lake,
Turkey. Reference [20] determined the concentrations of Pb,
Cd, Zn, Ni, Cu, Cr and Hg in liver and muscle of Labeorohita
and Ctenopharyngodonidella in the Lake of Bhopal, India. It
was showed that the concentrations of heavy metals in liver
were higher than muscle.
The mean estimated concentrations for Hg, Pb and Cd in
the present study were lower than international standards for
these metals as declare by [33] and [34]. Concentrations of As
in this study were higher than [35] (Table III). Muscle As
concentrations in the two fish exceed WHO limited,
representing a potential risk for human consumption in case
fish were eaten.
TABLE III: COMPARISON OF HG, CD, PB AND CU CONCENTRATIONS (MG/ KG)
IN MUSCLE OF BARBUSGRYPUS AND CAPOETACAPOETA WITH STANDARDS
(WORLD HEALTH ORGANIZATION, U.S. FOOD AND DRUG ADMINISTRATION,
MINISTRY OF AGRICULTURE, FISHERIES AND FOOD (UK), NATIONAL
HEALTH AND MEDICAL RESEARCH COUNCIL (AUSTRALIA), FOOD AND
AGRICULTURE ORGANIZATION) (MG/ KG)
Standards Hg Cd Pb As References
WHO 0.5 0.2 2 0.02 WHO, 1996
FDA 0.5 2 0.5 - Chen and Chen,
2001
UK (MAFF) 0.5 0.2 2 - MAFF, 1995
NHMRC 1 0.05 1.5 - Chen and Chen,
2001
B. grypus 0.072 0.120 0.248 0.138 This study
C. capoeta 0.063 0.111
Although rock weathering, atmosphere deposition and
phosphate mineral sources are the natural inputs of heavy
metals to water systems, different anthropogenic activities
such as discharges from electroplating and textile factories,
ship antifouling paints, agriculture runoff and vehicle
emissions all contribute to heavy metal pollution in marine
sediments [36]. These toxic elements consequently
accumulate in fish muscle, threatening human health through
the consumption of contaminated fish. Such non- essential
metals also poses the most harm since continuous exposure of
organisms to low concentrations of these metals may result in
their bioaccumulation, and their subsequent transfer to human
beings via the food chain [37]. In above investigations, fishes
have been successfully used as accurate indicators organisms
for environmental monitoring programmes because they
posses numerous advantages which include: 1) They are
typically present in all aquatic systems; 2) there is extensive
life- history and environmental response information
available for most species; 3) fish communities usually
include a range of species that represent a variety of trophic
levels and include of foods ob both aquatic and terrestrial
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TABLE II: THE CONCENTRATIONS OF HEAVY METALS IN MUSCLE OF
BARBUS GRYPUS AND CAPOETA CAPOETAIN HELEH RIVER (MG/ KG)
Fish Heavy metal muscle
mercury 0.072±0.004a
cadmium 0.120±0.022a
Barbusgrypus lead 0.248±0.037a
arsenic 0.138±0.025a
mercury 0.063±0.007a
Capoetacapoeta cadmium 0.111±0.056b
lead 0.219±0.072b
arsenic 0.115±0.038b
a. non-significant differences at P < 0.05. b: significant differences at P<
0.05.

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origin; 4) they are comparatively stable and therefore provide
a long- term record of environmental stress; 5) they contain
many life forms and functional guilds and thus are likely to
cover all components of aquatic systems affected by
anthropogenic disturbance; and 6) they are both sedentary and
mobile and thus reflect stressors within one areas as well as
providing scientists to give a broader assessment of effects
[38]. Therefore, the elucidation of heavy metal levels in the
fish species investigated in this study provided an indication
of the current environmental conditions of the Heleh River.
Metal concentrations measured in fish are directly or
indirectly influenced by a large set of biotic and abiotic
factors. Fish can uptake trace metals by two main routes,
either by absorption from water through the gill, and from
food absorbed through the digestive tract. According to the
literatures, metal bioaccumulation by fish and subsequent
distribution organs is greatly inter- specific. In additions,
many factors can influence metal uptake like sex, age, size,
reproductive cycle, swimming patterns, feeding behavior and
living environment [39].
Metal present in water show different bio- availabilities,
both for fish and their prey. Water chemistry features such as
dissolved and suspended organic carbon, pH, hardness and
alkalinity are important modified of metal bio- availabilities
and toxicity to aquatic organisms.
Many factors can be at the region of the difference between
fish species collected in the same site, e.g., diet, lifespan and
physiological characteristics. Several authors attribute the
interspecific differences observed in the metal burden of fish
tissues to variation in diet [40].
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Laleh Roomiani was born in Dezful, Iran on September
11, 1978. She has taken the PhD degree in fisheries
course from Islamic Azad University, Tehran Science
and Research Branch in Tehran in 2012. Her field of
study is aquaculture and food science. Roomiani is one
member of the Department of Fisheries, Agriculture
Faculty in Islamic Azad University in Ahvaz, Iran.