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Vol. 3: 1- 10
http://www.pvamu.edu/texged Prairie View A&M University, Texas, USA
According to the National Water inhabiting seven locations in the River Nile
Research Center (NWRC, 2000), the River during winter and summer seasons (2009).
Nile from Aswan to Al-Kanater Barrage Also the effect of the pollution of the River
receives wastewater discharge from 124 Nile on some biochemical parameters and
point source, of which 67 are agricultural enzyme activities in the muscles of fish were
drains and the remainders are industrial studied.
sources.
Bio-monitoring of hazardous 2. MATERIAL AND METHODS
substances in tissues of aquatic organisms 2.1 Sampling:
has been successfully applied during recent Samples of Oreochromis niloticus (L.)
years for heavy metals pollution (Lamas et fish were collected during winter (Joinery)
al., 2007). and summer (August) 2009 from seven
Fish are often at the top of the aquatic stations along the main channel of the River
food chain and metals are accumulated in it Nile at Upper Egypt covering different
to concentrations much time higher than that environmental conditions .The sampling
present in water and sediment. Fish can stations included Aswan reservoir, Chema,
absorb heavy metals through epithelial or Com Ombo, Naga Hammadi, Qus, El Menia
mucosal surface of the skin, gills and and El Hawamdia. The weights of the fish
gastrointestinal tract (Jovanovic et al., 2011) samples ranged from 77 to125g and the
.Fish in the River Nile was used as a lengths from15 to21.5 cm.
biological marker for its pollution .The 2.2 Determination of heavy metals
impact of heavy metals on fish affects A representative sample of 1g dry
directly or indirectly human health weight of each tissue (gills, intestine and
(NWQCU,1995). Oreochromis niloticus is muscles) was taken from 8 fish specimens of
one of the aquatic organisms affected by Oreochromis niloticus. The samples were
heavy metals, so it was frequently used as a digested according to the method described
metal biological marker in toxicological by Goldberg et al. (1993) in which
studies (Rashed, 2001 a&b). Studies from concentrated nitric and perchloric acids with
the field and laboratory experiments showed ratios of 5ml + 5ml were used in Teflon
that accumulation of heavy metals in fish beakers on a hot plate at 50 °C for about 5
tissues is mainly dependent on hours till complete decomposition of organic
concentrations of metals in water and matter. The digested solutions were cooled to
exposure period. The bioaccumulation of room temperature, filtered and diluted to a
heavy metals in the different fish tissues had final volume of 50 ml using deionized
been studied by several investigators (Gad distilled water. The concentrations of copper,
and Yacoub 2009; Cao et al., 2010; Malik et manganese, lead and zinc in the gills,
al., 2010; Jovanovic et al., 2011; Ebrahim intestine and muscles of O. niloticus were
and Taherianfard, 2011).Biochemical profile measured by SHIMADZU atomic absorption
in fish has proved to be a sensitive index for spectrophotometer model AA- 6800
evaluation of the fish metabolism under equipped with Flame Unit and auto-sampler
metallic stress .Studies proved that, fish SHIMADZU ASC- 6100 in Mediterranean
subjected to metals shoed reduced levels of Sea branch of National Institute of
proteins, lipids, ALT and AST activities in Oceanography & Fisheries. Results were
the muscles (Almeida et al ., 2001; Atli and expressed in mg/Kg dry weight of the tissue.
Canli, 2007; Mohamed and Gad, 2008). 2.3 Biochemical analysis:
The aim of the present study was to Samples from muscles of the studied
investigate the distribution of some heavy fish were subjected for calorimetric analysis
metals (Cu, Mn, Pb and Zn) in certain tissues of total proteins content (Henry, 1964)
(gills, intestine and muscles) of O. nilotucus (SPECTRUM kit, Egypt), total lipids
Amal M. Yacoub and Nahed S. Gad Accumulation of some heavy metals in Oreochromis niloticus 3
(Knight et al., 1972). ALT and AST stations of the River Nile during winter and
activities were calorimetrically assayed summer seasons 2009. The lowest
following the method of Reitman and accumulation levels of the tested metals were
Frankel (1957), using SPECTRUM kit. recorded in the muscle, while the highest
2.4 Statistical analysis: The data were ones were recorded in the intestine.
computed, expressed as means + standard 3.2 Copper (Cu):
error and statistically analyzed (Snedecor, Table (1) shows the concentrations
1962). of Cu in the studied tissues of O. niloticus
from the seven stations of River Nile. Cu
3. RESULTS accumulation in the studied tissues of fish
3.1 Metals Concentration in fish Tissues followed the order: intestine>gills>muscles
Tables (1-4) show the concentrations of the for all stations in winter and summer
analyzed metals in the tissues (gills, intestine seasons. The highest level of Cu in different
and muscles) of O. niloticus from seven fish organs was recorded in winter season.
Table 1: Concentrations of copper (mg/kg dry wt.) in different organs of O. niloticus collected from River Nile
during summer and winter (means +SD)
Gills Intestine Muscles
Stations
1 2 1 2 1 2
Aswan Reservoir 4.0+0.01 5+0.03. 7.3+0.01 9.7+0.03 2.1+0.01 14.8+0.03
4 Amal M. Yacoub and Nahed S. Gad Accumulation of some heavy metals in Oreochromis niloticus
Table 3: Concentrations of lead (mg/kg dry wt.)in different organs of O. niloticus collected from River Nile
during summer and winter (means +SD)
Stations Gills Intestine Muscles
1 2 1 2 1 2
Aswan Reservoir 13.8+0.1 10+0.3. 7+0.1 12+0.3 11.5+0.1 9.7+0.3
Chema 22.8+0.3 25+0.4 17+0.8 1 13.2+0.4 14+0.2 13+0.1
Com Ombo 15+0.3 15+0.2 14+0.3 4 18+0.2 20+0.1 12+0.4
Naga Hammadi 21+0.2 12+0.3 12.6+043 17+0.3 19+0.1 9+0.2
Qus 21+0.4 16+0.2 22+0.5 42+0.2 18+0.4 9+0.1
El-Menia 22+0.1 15+0.1 21+0.4 13.6+0.1 17+0.1 11+0.3
ElHawamdia 20+0.3 13+0.1 22+0.7 1 14+0.1 19+0.2 14+0.1
1-summer samples 2- winter samples
Table 4: Concentrations of zinc (mg/kg dry wt.) in different organs of O. niloticus collected from River Nile
during summer and winter (means +SD)
Stations Gills Intestine Muscles
1 2 1 2 1 2
Aswan Reservoir 62.8+.2.0 13+.3.0 30+1.8 22+.3.0 44+ 1 45+ 3.8
Chema 72+.3.0 68+.4.5 94+8. 1 87.2+4.4 70+.2 62+.1.3
Com Ombo 81+.3.8 13+.2.2 89+3. 4 47+.2.5 53+.1.5 11+4.0
Naga Hammadi 84+.2.6 59+ 3.6 70.6+3.5 78+.3.2 66+1.7 32+2.2
Qus 99+ 6.0 76+2.7 94+ 5.6 122+2.4 62+.43 37+.1.2
El-Menia 72+ 5.0 52+1.4 77+ 4.6 63.6+1.4 53+1.2 62+3.2
El Hawamdia 74+.3.0 25+.1.8 40 +7. 1 22+1.5 60+.2.2 18 +1.5
1-summer samples 2- winter samples
Table 5: Total proteins and lipids contents in muscles of Oreochromis niloticus from River Nile (means +SD)
Stations Total proteins Total lipids
1 2 1 2
control 15.9+.0.07 16+.0.6 0.67+1.8 0.70+0.5
Aswan reservoir 13.+ 0.05 14+3.2 0.82+ 6.3 0.79+0.06
(-16.9) (-12.5) (-22.3) (+12.8)
Chema 9.6+0.04 8 +.0.03 0.51+0.02 0.50+0.04
(-39.6)* (-50.0)** (-23.8) (-28)*
Com Ombo 7.4+.0.03 6.0+.0.02 0.64+3. 4 0.45+0.06
(-53.4)** (-62.5)** (-4.4) (-35.7)*
Naga Hammadi 7.8+.0.02 7.5+0.03 0.53+0.0.5 0.46+.0.04
(-50.9)** (-53.1)** (-20.8) (-34.2)*
Qus 8.1+0.04 10.1+0.02 0.8+0.05 0.82+0.05
(-49.0)** (-37.5)* (+19.4) (-14.28)
El-Menia 9.6+0.06 4.2+0.05 0.86+0.07 0.32+00.06
(-39.6)* (-73.7)** (-4.0) (-54.2)**
ElHawamdia 8.5+0.06 6.3+.0.04 0.62+0.01 0.55+0.03
(-46.5)** (60.6)** (-5.0) (-21.4)
Data are represented as means +S.E of 6 fish
*Significantly different from control (P<0.05)
**Highly significant different from control (P<0.01)
Data in parentheses are percent of change from control
1-summer samples 2- winter samples
Amal M. Yacoub and Nahed S. Gad Accumulation of some heavy metals in Oreochromis niloticus 5
The results indicated that there was a Table (6) presents the changes in the
highly significant (P<0.01) decrease in the activities of ALT and AST in the muscle of
total proteins in muscle of the studied fish O. niloticus collected from River Nile during
compared to the control except in station (1) winter and summer seasons. The results
where, there was a decrease in total proteins indicated that there was a highly significant
in muscles of O. niloticus but not statically (P<0.01) decrease in the activities of ALT
significant during summer season. On the and AST in the muscle of the studied fish
other, hand there was a highly significant compared to the control during winter season
decrease in the total lipids in the muscles of except in station (1). In summer season the
the studied fish during winter season activities of ALT and AST were decreased
(P<0.01). but statistically not significant.
Table 6: Alanine amino transferase (ALT) and aspartateaminotranseferase (AST) in muscles of Oreochromis
niloticus from River Nile (means+SD)
6 Amal M. Yacoub and Nahed S. Gad Accumulation of some heavy metals in Oreochromis niloticus
The maximum permissible level (MPL) accumulation in gills that cause a structural
of zinc is 50 ug/g dry wt. according to damage and a reduction in oxygen
Australian NHMRC (Bebbington et al., consumption causing sharp reduction in the
1977) and 40 ug/g dry wt. according to Food metabolic rate of fish and consequently
and Agriculture Organization (FAO, 1983). decrease protein contents in tissues.
The mean values of zinc in the gills, intestine Moreover, decreased tissues protein in fish
and muscles of O. niloticus were living in polluted environment may be a
significantly higher than (MPL) except in result of decreased insulin level caused by
fish muscles in winter. The concentrations of metal toxicity (Zaghloul, 2001). Insulin is
heavy metals (Cu, Mn, Pb and Zn) varied known to have profound effects on the
according to season, locality and tissue type proteogenic pathways in fish. It stimulates
(Yacoub, 2007). Metal accumulation in the inward cellular transport of amino acids,
tissues of fish is dependent upon particularly in muscle, leading to
environmental factors such as temperature, intracellular accumulation of amino acid
size and age of fish and processes of with subsequent decrease of protein contents
biotransformation and excretion (Zhou et al., (Reda et al., 2002).
2001). Concerning the effects of different
Studies proved that, fish subjected to pollutant in the River Nile on muscle lipids
metals shoed reduced levels of proteins, contents of the studied fish the obtained data
lipids, ALT and AST activities in the showed highly significant decrease during
muscles (Almeida et al., 2001; Atli and winter season but in summer the decrease in
Canli, 2007; Mohamed and Gad, 2008). total lipids in muscles was statistically non
In various fish species, proteins are of significant. The decrease in total lipids
importance as structural compounds, contents in muscle of the studied fish may
biocatalysts and hormones for control of be due to secretion of catecholamine and
growth and differentiations. So variation in corticosteroids in the blood after the toxicant
fish proteins could be used as bioindicator stress that produces an enhanced metabolic
for monitoring physiological status of the rate which in turn reduces the metabolic
tested fish (Begum and vijayaraghavan reserves (as proteins and lipids) (Marie,
1996). 1994). Also, the decrease in muscles total
In the present study, the examined O. lipids may be due to the use of energy-rich
niloticus collected from different stations of lipids for energy production during toxic
the River Nile exhibited lowered protein stress as previously reported by Sancho et
value in muscles during winter season, which al., (1998). The decreased lipids content
was suggested as a metabolic adaptation to observed in the present investigation agrees
food shortage in the environment (White et is with that recorded in muscle of fish
al.,1986). During this period of inadequate exposed to different pollutants (Chandra et
food supply, energy required for metabolic al., 2004 and Blaner et al., 2005).
maintenance may be provided from The environmental pollution in the
utilization of protein reserves which mainly River Nile induced a marked decrease in the
accumulate in the muscle tissues (Haggag et muscle ALT and AST activities. The results
al., 1999). Besides, this protein depletion indicate that under the influence of different
could be attributed to change in the water heavy metals or in a state of stress, the
quality of River Nile as a result of the damage of muscle tissues may occur with
discharged effluents from different sources, concomitant liberation of transaminase into
including hydrocarbons found in sewage the circulation (Allen, 1995).The decreased
wastes and heavy metals in the industrial and activities of ALT and AST indicate
agricultural drainage. (Zaghloul, 2000). This disturbance in the structure and integrity of
may be explained as the exposure to metals cell organelles, like endoplasmic reticulum
(as Cu and Zn) may lead to high and membrane transport system (Humtsoe et
8 Amal M. Yacoub and Nahed S. Gad Accumulation of some heavy metals in Oreochromis niloticus
al., 2007). Marie (1994) stated that the exposure and metabolic responses of the Nile
reduction in ALT and AST activities in fish tilapia Oreochromis niloticus. Environ
exposed to metals could be attributed to the .Pollut., 114:169-175.
high accumulation of metals in fish tissues. Atli,G. and Canli, M. (2007).Enzymatic
responses to metal exposures in a freshwater
Therefore, the depletion of enzymes in the
Oreochromis niloticus. Comp. Biochem.
fish tissues observed in the present study can Physiol. C Toxicol. Pharmacol., 145:282-
be attributed to the increased metals 287.
accumulation in the tissues. The reduction in Australian NH and MRC (1987): Food Standards
the muscle ALT and AST activities in the code – 1987. Australian Government
fish exposed to various pollutants and heavy Publishing Service, Canberra.
metals, in particular have been reported by Bebbington, G. N.; Mackey, N.J.; Chvoike, R.;
Rao (2006); Humtsoe et al. (2007) and William, R. J.; Dunn, A. and Auty, E. H.
Mohamed and Gad (2008). (1977). Heavy metals (selenium and arsenic)
In conclusion, the results of the present in nine species of Australian Commercial
study revealed that the abundance of Fish. Aust. J. Mar. Freshwater Res., 28:
277- 280.
heavy metals in fish organs followed the
Begum, G. and Vijayaraghavan, S. (1996).
order: Mn>Zn>Pb and Cu. The highest Alterations in protein metabolisms of muscle
accumulation of the heavy metals was tissues in the fish Clarias batrachus by
recorded in the intestine and the lowest commercial grade Dimethioate. Bullet.
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below the maximum permissible limit, while Blaner, C.; Curitis, M. and Chan, H. (2005).
Mn, Pb and Zn in the muscles exceeded the Growth, nutritional composition and
permissible limit. It was found that the hematology of Artic chair
environmental pollution in the River Nile (Salvelinusalpinus) exposed to toxaphene
induced significant changes in the proteins and tapeworm (Diphyllobothrium
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