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O A RIGINAL
3857
Journal of Applied Sciences Research, 9(6): 3857-3872, 2013
ISSN 1819-544X
This is a refereed journal and all articles are professionally screened and reviewed
ORIGINAL ARTICLES
Histopathological and muscle composition studies on Tilapia zillii in relation to water
quality of Lake Qarun, Egypt
1
Safaa I. Tayel; 2Seham A.Ibrahim and 1Soaad A. Mahmoud
1
National Institute of Oceanography and Fisheries, Inland Water and Aquaculture Branch, El-Kanater
El-Khairya Fish Research Station, Egypt
2
Department of Zoology, Faculty of Science, Benha University, Egypt
ABSTRACT
This study was conducted during spring 2011 to investigate the impact of water quality changes in Lake
Qarun on the histopathology and muscle composition of Tilapia zillii fish living in this lake . Physico-chemical
parameters i.e. air and water temperatures, electrical conductivity, pH, salinity, dissolved oxygen, biochemical
oxygen demand, chemical oxygen demand, ammonia, nitrite, nitrate, phosphate and silicate were determined.
Histopathology of liver, spleen, gills and muscles as well as the muscle composition including, water, protein,
lipid, ash and carbohydrates contents of the fish were done .The results showed that changes in water quality
had negative impact on histopathology of selected organs and muscle composition of the studied fish. The
resulted damage were related to the large amounts of contaminated drainage water from Faiyoum Province,
which consequently may affect fish production and human health. So it is necessary to treat the drainage water
before discharging into the lake.
Key words: Water pollution, Fish, Histopathology, Muscle composition, Lake Qarun , Egypt
Introduction
Lake Qarun is an inland lake occupies the lowest part of El- Faiyoum depression (Yacoub et al., 2008). It is
a closed system acts as a reservoir for agricultural and sewage drainage water which received from El-Faiyoum
province (Ali and Abdel Satar, 2005). Its water salinity increases progressively, which affects greatly the lake
biota, in addition, the exacerbation of eutrophication of the lakeʼs water that caused by the nutrient load from
the agricultural drainage water (Sabae and Ali, 2004)
The lake receives the agricultural and sewage drainage water through a system of twelve drains, most of the
drainage water reaches the lake by two main drains, El- Batts and El- Wadi, whereas there are minor drains
poured its drainage water into the lake by means of hydraulic pumps but in small amounts (Gaber and Gaber,
2006). The minor drains are recently connected with a larger drain, namely Dayer El-Birka, which transfers a
part of wastewater to the lake by pumping stations (Authman and Abbas, 2007).
The lake receives annually about 450 million cubic meters of agricultural drainage water which
approximately balances the amount of lake water lost annually by evaporation, leading to progressive increase
of salinity and detrimental effects to the lake environment, e.g. its fauna and flora (Yacoub et al., 2008).
The Egyptian Company for Salts and Mineral (EMISAL) located in the southern coast of the lake, was
cutoff from the lake and divided into number of ponds, to concentrate the lake water as much as 10 times of its
original salinity. The effluents of EMISAL brine water discharged also into the lake affecting the ecosystem
condition.
Because of lake Qarun is a closed ecosystem, and as a result of extensive evaporation of water, the
accumulation of chemical pollutants (heavy metals, pesticides and other pollutants) is expected to increase
annually in all its components (e.g. water and fish) and change their quality and affect their aquatic life
(Mansour and Sidky, 2003).
Previous studies reported that Lake Qarun components are polluted with heavy metals (Ali and Fishar,
2005), solid and nutrients (Gupta and Abd El-Hamid, 2003) and with a wide variety of pesticides (e.g. lindane,
aldrin, some DDT analogues malathion) (Ali et al., 2008). Moreover, a remarkable increase in the bacterial
indicators of sewage pollution (total coliform, faecal coliforms and faecal streptococci) in the lake was recorded
(Sabae and Rabeh, 2000). Histopathological alterations can be used as indicators for the effects of various
anthropogenic pollutants on organisms and are a reflection of the overall health of the entire population in the
ecosystem (Saad et al., 2011). These histopathological biomarkers are closely related to other biomarkers of
stress since many pollutants have to undergo metabolic activation in order to be able to provoke cellular change
Corresponding Author: Safaa I. Tayel, National Institute of Oceanography and Fisheries, Inland Water
andAquaculture Branch, El-Kanater El-Khairya Fish Research Station, Egypt
E-mail: [email protected]
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J. Appl. Sci. Res., 9(6): 3857-3872, 2013
in the affected organism. Previous studies reported that the exposure of fish to pollutants (agricultural, industrial
and sewage) resulted in several pathological alterations in different tissues of fish (Saad et al., 2012).
Histopathological changes were observed in the muscles of fish as a result of exposure to different toxicants
(Abbas and Ali, 2007).
Tilapia species are the major protein in many of developing countries. They are considered to be a good fish
having a white flesh with no intramuscular bones and it is considered as a good source of animal protein in the
tropic and sub-tropic regions (Tayel, 2003). The principle constituents of the fish flesh which are water ,protein
,lipid, carbohydrates and ash contents are greatly affected by water quality such as temperature, pH and salinity
(Tayel, 2007).
The additional information still needed to provide a database for the ecological status of lake Qarun that
helps the policy markers to take effective decision for proper management of lake.
This study aimed to investigate the impact of the environmental pollution of lake Qarun on histological
structures of liver, spleen, gills and muscles and biochemical composition of muscles of a commercially
important fish ; Tilapia zillii that inhabiting this lake.
Material And Methods
Area of study:
Lake Qarun is one of the enclosed saline lakes in Egypt . It is located in the western desert part of Faiyoum
depression and lies 83 km southwest of Cairo (Fig. 1). The lake is located between longitudes of 30º 24 & 30º
49 E and latitude of 29 º 24 &29 º 33 N. It is bordered from its northern side by desert and by cultivated land
from its south and southeastern side (Abou El-Gheit et al., 2012). The lake receives the agricultural drainage
water from the surrounding cultivated land. The drainage water reaches the lake by two huge drains; El-Batts
drain (at the northeast corner) and El-Wadi drain (near mid – point of the southern shore).
Field observation:
The water samples from Qarun lake were collected during spring 2011 to measure air and water
temperature (oC) by a dry mercury thermometer ,as well as electrical conductivity, by using Hydrolab, Model
(Multo 3401/SET). pH by Orion Research Ion Analyzer 399; A h meter.
Laboratory analysis:
Water samples:
Another water sample was kept in one liter polyethylene bottle in ice box to be analyzed in the laboratory.
Salinity was determined according to the standard method described by the American Public Health Association
(APHA, 1985). The dissolved oxygen content analysis was performed by azid modification and biological
oxygen demand by incubation 5 day methods. Chemical oxygen demand by potassium dichromate oxidation.
Concentration of ammonia, nitrite, nitrate, phosphate and silicate were determined using the colorimetric
techniques according to the method described by APHA (2002).
Fish samples:
Histopathological studies:
Liver, spleen, gills and muscles of Tilapia zillii fish were carefully removed then fixed in 10% formalin,
dehydrated in ascending grades of alcohol and cleared in xylene. The fixed tissue were embedded in paraffin
wax and sections of 5 microns were cut, using Euromex Holland microtome. Sections were stained with Harris
Hematoxylin and Eosin method; (cited by Saad et al., 2012). Consequently, these sections were examined
microscopically and their photos were taken by microscopic camera. Finally these sections compared by those
obtained from the controlled fish collected from El-Kanater El-Khairya fish research station.
Biochemical Composition:
The scales, skin and bones of fish specimens were carefully removed. Flesh sample was taken from the
mid-dorsal region of each specimen.
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J. Appl. Sci. Res., 9(6): 3857-3872, 2013
Water content:
Fish flesh sample (about 3 g), was transferred to a tarred weighing beaker (weighted accurately) and placed
in a drying oven thermostatically regulated at 105 ºC overnight. The loss in weight was taken as equivalent to
weight of water content (Sidwell et al., 1970).
Protein content:
Crude protein was determined by total nitrogen estimated using a " modified micro-kjeldahl's method (ElAggan, 1982). Complete digestion of dried samples should be done. Liberated ammonia is distilled into 2 %
boric acid, using bromocresol green methyl red as indicator. Crude protein was calculated by multiplying total
nitrogen by factor 6.25 (for fish).
Lipids content:
Dried samples (each about 2 g) were individually used for lipid extraction by chloroform as a solvent for 6
hours in a soxlet apparatus at 65 ºC. The extracted lipid were estimated by loss in weight before and after
extraction of chloroform, and its percentage was calculated (AOAC, 1980).
Ash content:
Ash was determined by igniting the dried sample of known weight (about 0.5 gm) using silica crucible, in a
muffle furnace at about 800oC for about 6 hours. After cooling in a dessicator, the ash was weighted and its
percentage was calculated (Sidwell et al., 1970).
Carbohydrate content:
Carbohydrate content was determined by calculation according to following equation:
Carbohydrate = 100 – ( Protein content + Lipids content + Ash content ) .
Statistical analysis:
The comparison of means ± SE ( Standard Errors ) was tested for significance using one –way ANOVA
analysis and Duncan΄s multiple range tests . The statistical analysis were calculated, using the computer
program of SPSS Inc. (2001 version 10.0 for Windows ) at 0.5 significance level.
Fig. 1: Map of Lake Qarun.
Results and Discussion
I-Water quality:
The obtained values of water analysis including air and water temperatures, electrical conductivity, pH,
salinity, dissolved oxygen, biochemical oxygen demand, chemical oxygen demand, ammonia, nitrite, nitrate,
phosphate and silicate and their permissible limits according to WHO (1993) are shown in Table (1).
Air and water temperatures:
They are critical control parameters in aquatic system and they are key parameters which influences the
physical, chemical and biological transformation in the aquatic environment (Delince, 1992; Mahmoud et al.,
2008).
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J. Appl. Sci. Res., 9(6): 3857-3872, 2013
The obtained values of air and water temperature ranged from 20.9 ºC to 23.3 ºC and from 18.8 ºC to 20.9
ºC, respectively. The changes of water temperature may depend on the variations in meteorological condition,
time of sampling, air temperature , back radiation and latent heat of evaporation and seasons as recorded by
Awad (1993) and Mahmoud et al. (2008).
Electrical conductivity:
The obtained values of electrical conductivity varied between 24368 μmohs/cm and 37650 μmohs/cm; these
values are higher than those of permissible limits of WHO (1993). The decreasing in EC values may be
attributed to the uptake of dissolved salts by phytoplankton as recorded by Ahmed (2007) . While the increasing
in EC values may be due to the high content of dissolved ions and cations as cited by Saad et al. (2011) or may
be attributed to evaporation rate, the intrusion of drainage water and consumption of lake salts by EMISAL
Company as mentioned by Abdel-Satar et al. (2010).
pH value:
The pH of natural water affects on biological and chemical reactions, controls the solubility of metal ions
and affects on natural aquatic life (Mahmoud et al., 2008). The desirable pH for fresh water is in the range of
6.5-9.0 for aquatic life (Chin, 2000). The obtained values of pH in the present study ranged from 6.5 -7.4 .These
values within the range of permissible limits of WHO (1993). The consumption of DO by algae and
phytoplankton leads to increasing of CO2 that leads to decreasing in pH values as recorded by Abou El-Gheit
et al. (2012).
Salinity:
The obtained values of salinity ranged between 18.5o % and 27.3o % ; these values are within the range
obtained by Abou El-Gheit et al.( 2012) , who revealed this increasing to the evaporation rates , the intrusion of
drainage water and consumption of lake salts by EMISAL company as mentioned by Abdel-Satar et al. (2010).
Dissolved oxygen (DO):
Dissolved oxygen (DO) is of fundamental importance to the life and health of aquatic organisms. Fishes
and other aquatic organisms depend for their respiration on the DO content in water body. The distribution of
DO is affected by the solubility of many inorganic nutrients (Mahmoud, 2002).
In the present study, dissolved oxygen values varied from 4.4 to 7.96 mg/l , these values are lower than
those of permissible limits of WHO (1993) . The decrease in dissolved oxygen concentration may be due to the
fall in water temperature and phytoplankton blooming (Konsowa, 2007 ; AbouEl-Gheit et al., 2012). Also, the
depletion in DO may be due to dissolved oxygen exhaustion for oxidation of huge organic matter discharged
into the lake; this observation agrees with that obtained by Saad et al. (2011).
Biochemical oxygen demand (BOD):
Biochemical oxygen demand (BOD) is the amount of dissolved oxygen which used to decompose the
organic matter in the water . It depends on several factors such as : temperature, concentration of organic matter
and density of phytoplankton. Also it increases by increasing the chemical oxygen demand ( Tayel,2003).The
BOD test is the mostly useful method in estimating the amount of biodegradable organic matter present in the
aquatic environment(Siliem,1993).
The obtained values of BOD ranged between 0.83 and 2.98 mg/l . These values are lower than the
permissible limits of WHO (1993) and those obtained by Abou El-Gheit et al. (2012). The lowest value of BOD
may be due to the lower photosynthetic activity and abundance of phytoplankton as cited by Ahmed (2007) .
While the high value of BOD may be attributed to the presence of high load of wastes discharged (agricultural
and sewage) into the lake as recorded by Saad et al. (2011).
Chemical oxygen demand (COD):
The chemical oxygen demand (COD) is the total amount of oxygen required to oxidize all the organic
matter completely to CO2 and H2O. COD test is used to assess the degree of pollution in the area under
investigation (Ahmed, 2007).
In present study, the values of COD ranged from 48.5 to 135.2 mg/l.These are values higher than the
permissible limits of WHO (1993) and those obtained by Abou El-Gheit et al. (2012) for the same lake. On the
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J. Appl. Sci. Res., 9(6): 3857-3872, 2013
other hand, these values are higher than the obtained values of DO and BOD . The high values of COD may be
due to high amount of organic matter in waste water or low capacity of water for self- purification as recorded
by Tayel (2003).
Ammonia (NH3):
Ammonia is considered one of the most important parameters that must be studied in pollution of water
ecosystem (Emerson et al., 1975). The term ammonia includes the non-ionized (NH3) and ionized (NH4)
species (Mahmoud, 2002) . The increase in the concentration of ammonia (NH3) leads to toxicity and mortality
of fishes and other aquatic organisms but ammonium ions (NH4-) have no effect on the aquatic organisms or
fishes (Siliem, 1993 and Ahmed, 2007). So, ammonia in water is an indicator of possible bacterial, sewage and
animal waste pollution (Tayel, 2003). Natural levels in ground and surface water are usually below 0.2 mg/l
(WHO, 1993).
Ammonia and nitrogen concentrations more than 1 mg/l have been given as indicator of organic pollution
and can be toxic to aquatic species if they are higher than 2.5mg/l as cited by Reid (1961) ; Siliem (1984) and
Saad et al. (2011), who added that, increasing toxicity of ammonia is attributed to the low dissolved oxygen.
The present results declared that ammonia accounted for the major proportion of total soluble inorganic
nitrogen. The values of ammonia ranged from 760 μg/l to 1278 μg/l; these values are within the range obtained
by Abou El-Gheit et al. (2012) and higher than the permissible limits of WHO (1993) (50-500 μg/l). The high
values of ammonia may be due to temperature (Saad et al., 2011), agricultural and sewage wastes (Abdel-Satar
et al., 2010) discharged into lake Qarun from surrounding cultivated land and two huge drain El-Batts and ElWadi. While the low values of ammonia are rarelated to the decrease in biological activities of aquatic
organisms and nitrification occurs in the water column (Abou El-Gheit et al., 2012 ). Oxidation of ammonia to
NO2 or NO3 is a subject of much ecological uncertainly as cited by Mahmoud et al. 2008.
Nitrite (NO2):
The obtained values of nitrite ranged between 1.36 μg/l and 2.6 μg/l ; these values are within the range
obtained by Abou El-Gheit et al. (2012). The low values of nitrite might be attributed to fast conversion of
nitrite by nitrobacteria to nitrate (Tayel, 2007). However, the high nitrite level might be attributed to
decomposition of organic matter present in the waste water where nitrosomonas bacteria oxidize ammonia to
nitrite by denitrification (Saad et al., 2011).
Nitrate (NO3):
The values of nitrate fluctuated within the range of 34.6 μg/l to 50.2 μg/l These results are within the range
obtained by Abou El-Gheit et al. (2012) and lower than the permissible limits of WHO (1993). Nitrate showed
high values than the corresponding values of nitrite due to the fast conversion of NO2- to NO3- ions by nitrifying
bacteria (Abdel-Satar et al., 2010) . The low values in nitrate concentration might be due to uptake of nitrate by
phytoplankton and its reduction by denitrifying bacteria and biological denitrification (Abdo, 2002).
Phosphate (PO4):
Phosphorus that enters the aquatic system through anthropogenic sources , e.g. fertilizer-runoff, potentially,
could be incorporated into either inorganic or organic fraction (Abou El-Gheit et al., 2012). Once phosphorus
accumulated within a lake , it can be cycled through the water column and promote algal blooms indefinitely
(Edwards and Withers, 2008). In the present study phosphates ranged between 79 and 168 μg/l. These values
showed lower rates than that measured in the same lake by Abdel-Satar et al. (2010) and permissible limits
(WHO, 1993). This reflects the indirect negative effect of algal blooming on the food web by decreasing the
amount of edible phytoplankton that zooplankton and other primary consumers need to survive on ( NOAA,
2009).
Silicates (SiO2):
The obtained values of silicates in the present study were ranged from 6.85 to 9.62 mg/l. These values are
lower than permissible limits of WHO (1993) and that of Abou El-Gheit et al. (2012).
The Pearson correlation coefficient matrix (r) between different water quality parameters of the investigated
area (Table 2 ), demonstrated some significant positive and negative correlations.. It was found that, there is a
highly significant positive correlation (r= 0.999, P<0.01) between air and water temperatures (i.e. the water
temperature followed that of air temperature). Air temperature showed positive correlations with pH, Phosphate
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J. Appl. Sci. Res., 9(6): 3857-3872, 2013
and Silicate (r= 0.934, 0.937 and 0.756, P>0.05, respectively) (i.e. pH, phosphate and silicate values increase
with increasing the air temperature and vice versa). Air temperature showed highly significant negative
correlation (r= -0.994, P<0.01) with nitrite (i.e. nitrite values decreased with increasing of air temperature). Air
temperature showed significant negative correlations with COD, BOD and salinity (r=-0.976, -0.978 and -0.970,
P>0.05, respectively) (i.e. the COD, BOD and salinity values decreased with increasing of air temperature).
DO showed negative correlations with air and water temperatures (r= -0.679 and -0.642, P<0.05,
respectively ; i.e. the DO values decreased with increasing of temperatures.
Histopathological studies:
Histology has been used as a test for evaluating toxic effects of water pollutants in fish (EIFAC, 1983;
Murty, 1986 and Tayel et al., 2007). Results from histological studies are useful in establishing water quality
criteria ( FAO, 1981 and Mahmoud and El-Naggar, 2007).
Liver:
Liver of fish is responsible for the digestion, filtration and storage of glucose (El-Naggar et al., 2009). It is
found in the anterior part of the body cavity as a brownish red mass (Yacoub et al., 2008). The liver also
produces many enzymes that stored in the gall bladder (Tayel et al., 2008). These enzymes assist in the
breakdown of food (Ahmed, 2007). Generally, the liver is consider as the principal organ of detoxification in
vertebrates and particularly in fish (Tayel, 2003). Meanwhile, fish liver is a good indicator of aquatic
environmental pollution, where one of the important functions of the liver is to clean of any poisons or
pollutants from the blood coming from the intestine (El-Naggar et al., 2009).
In present study, liver of controlled Tilapia zillii fish was almost uniform in appearance as well as it was
soft in consisty and uniformly dark red color. It is enclosed within a fibroconnective tissue capsule. While these
liver cells sample histologically appear forming a meshwork and they are arranged in a definite cord. Like
pattern around well defined sinusoid leading to central vein (Fig. 2 ). The hepatic cells appeared as polyhedral
cells with central nuclei. Blood flows from branches of hepatic portal vein and hepatic artery through the
sinusoids to central veins which empty into the hepatic vein, this structure also explained by Groman (1982) and
El-Naggar et al. (2009).
Liver samples obtained from Tilapia zillii fish inhabiting the water of lake Qarun suffered from many
pathological alterations. Morphologically, the liver revealed pale color and enlargement appearance. While the
observed histopathological changes of liver were summarized in fatty degeneration, degeneration and necrosis
in hepatic cells and dilation, stagnant blood, hemorrhage and hemosidrin in blood vessels (Figs. 3-7) .These
results agree with those obtained by Yacoub et al. (2008); El-Naggar et al. (2009) and AboEl-Gheit et al.
(2012). The present study suggests a strong link between water quality changes and observed liver lesions, these
lesions which may be due to oxygen depletion (Gaber and Gaber, 2006 ; Ibrahim, 2007), parasitic infection( ElNaggar et al., 2009), elevation in ammonia (Yacoub et al., 2008), heavy metals (Mohamed, 2009). Also
hemorrahge caused by bacteria present in sewage water as recorded by Fouze et al. (1995); Tayel (2003) and
Saad et al., 2011 who added that necrosis may also caused by bacteria and toxins secreted by micro-organism in
sewage water. However, accumulation of hemosidrin in liver cells may be due to rapid and continuous
destruction of erythrocytes as recorded by Ibrahim and Mahmoud, (2005) and Tayel et al. (2008).
Spleen:
Spleen of fish is an important member of the body's immune and lymphatic system (Tayel et al., 2007). It is
a hematopoeitic tissue, which form the red blood cells and found as a small red mass ( El-Naggar et al., 2009).
Its functions are filtration of blood, producing and storage of red blood cells, removing old and abnormal
erythrocytes and producing antibodies against blood born antigens ( Saad et al., 2011).
In the present work, spleen of the controlled Tilapia zillii fish almost enclosed within a capsule and
consisted of lymphatic laden area, white pulp. The surrounding pink areas were the red pulp which contains
large numbers of free cells and sinusoids. Endothelial cells of the sinusoids were not in contact but separated by
slit like spaces. Red blood cells were broken in the red pulp. Macrophages may contain pigments from broken
down erythrocytes (Hemosidrin) (Fig. 8); this structure agree with that obtained by Groman (1982) and Saad et
al. (2011).
Spleen of Tilapia zillii fish inhabiting the water of lake Qarun suffered from many pathological alterations.
Morphologically, the spleen revealed dark color and enlarged shape , this observation agree with that noticed by
Tayel et al. (2007) who revealed this observation to viruses present in the sewage water. The observed
histological changes of spleen cells were summarized in hemorrhage, hemolysis, hemosidrin, necrosis and
degeneration in spleenic tissues (Figs. 9-13). The observed malformation come in agreement with those
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J. Appl. Sci. Res., 9(6): 3857-3872, 2013
obtained by Tayel et al. (2007&2008); Ibrahim et al. (2009) and Saad et al. (2011). These alterations may be
due to viral and parasitic microbes and trace metals which were accumulated in the spleen from polluted water.
Such explanation were recorded by Mazhar et al. (1986); Nigaga (2003), Tayel et al. (2008) and Saad et al.
(2011) . Also the hemosidrin formation (accumulation) may be attributed to the increase in iron content as
reported by Haggag et al. (1993) ; Tayel et al. (2008) and Saad et al. (2011).
Gills:
The fish gills are a multifunctional organ involved in respiration and homeostatic activities such as
osmoregulation, metabolism, circulation of hormones, nitrogen excretion and acid base balance (Haaparanta et
al., 1997 and Yacoub et al., 2008). They are among the most delicate structures of the teleost body which have
an external location so they are subjected to damage by irritant whether dissolved or suspended in the water
(Naidu and Ranmurthi, 1989 and Ibrahim and Tayel, 2005). External irritant are the most frequent causes of
significant gill pathological changes and retard the respiratory function of the organ by reducing its surface area
(Zaki &Saad, 1987 and Saad et al., 2011).
In present study, gill of the controlled Tilapia zillii fish was almost consist of several cartilaginous arches .
Each arch bears pairs of processes "Primary lamellae " which serve more for support of secondary lamellae than
for respiration. From the side of which radiate a thin expansions, the so called "Secondary lamellae"(Fig. 14).
Each secondary lamellae contains a thin walled gill sinusoid which performs the function of respiration,
excretion and osmoregulation and surrounded by "Pillar cells".The secondary lamellae are covered with
squamous " Epithelial cells " made up of specialized and undifferentiated cells. Among these cells the " Mucous
" and " Chloride " cells are found. The chloride cells (ioncytes), present in euryhaline species only and are the
primary osmoregulatory cells of fish. They are chiefly found at the base of both types of lamellae. This
description was also reported by Yacoub (1999) ; Tayel (2003) and Ibrahim et al. (2009).
Gills of Tilapia zillii fish living in the water of investigated area suffered from many pathological alteration.
Morphologically, gills lamellae revealed dark color and filaments existing covered with mucin, These
observations agree with that obtained by Yacoub et al. (2008) and Ibrahim et al. (2009). Histopathological
changes of gills were summarized in degeneration, necrosis, hemorrhage, hemosidrin, and separation in primary
and secondary lamellae (Figs. 15-19). These results are similar to those obtained by Ibrahim and Tayel (2005) ;
Yacoub et al. (2008) ; Ibrahim et al. 2009 and Abou-El-Gheit et al. (2012), who revealed these malformation
in gills to increase of ammonia and heavy metals, pH change, oxygen depletion occurrence of bacteria, microorganisms and parasites with increasing in turbidity in water polluted by sewage and agricultural discharge in
lake Qarun, respectively.
Fernandes and Mazon (2003) suggested that the bacteria produce in extracellular hyperplasia inducing
factor and they added that these pathological changes may be a reaction to toxicants intake or an adaptive
response to prevent the entry of the pollutants through the gill surface ( Abou El-Gheit et al., 2012).
Muscles:
The muscular system constitutes the largest portion of the teleost body. Its function in overall body are
locomotion, coordinated movement of skeletal elements, pumping of blood and peristaltic constriction of
visceral organ and their related structures (El-Serafy et al., 2005 and Tayel, 2007).
Recently, the epidermis have been shown to contain naturally bacteriolytic substances, and to contain a
continuous reacting immune system. Thus the affected epidermis must be loss its protective function as well as
its efficiency as an osmotic barrier (Tayel, 2003). If fish have epithelial lesion in polluted water, they would
probably be invaded by micro- organisms. Also, vitamin D-deficient fish cause severe epidermal pathology
resulting in skin and underlying musculature destruction (Taveekijakarn et al., 1996 and Tayel, 2003). In the
present study, skin of control fish consists of epidermal, dermal and hypodermal layers. The skin covers the
muscles layer which is composed chiefly of segmental myomeres. Each myomere is regarded as muscle and its
fibers are parallel the long axis of the body (Figs.20&21); this structure is agree with that cited by Bayomy &
Tayel (2007) and Yacoub et al. (2008). Skin and muscles of Tilapia zillii fish collected from investigated area
suffered from many pathological alteration. Morphologically, skin and muscle of Tilapia zillii fish inhabiting the
water of investigated area showed destroyed fin and loss of scales in some spacimens,that observation agree
with Tayel (2003 & 2007); Ahmed (2007) and Saad et al. (2012).
Histologically , alteration of skin and muscles of Tilapia zillii fish collected from investigated area were
summarized in hemorrahge ,hemosidrin , Fatty degeneration and necrosis in connective tissue of hypodermal
layer as well as degeneration, necrosis and edema in muscle fiber layer (Figs.22-26) . These results agree with
those obtained by El-Serafy et al. (2005); Sitohy et al. (2006); Ahmed (2007) ; Yacoub et al. (2008) and Saad et
al. (2012).
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J. Appl. Sci. Res., 9(6): 3857-3872, 2013
These alterations in skin and muscles may be attributed to inorganic fertilizers (Mahmoud,2002), ammonia
(Tayel, 2003), heavy metals (Sitohy et al., 2006; Ibrahim, 2007 and Yacoub et al., 2008), parasitic infection
(Mahmoud and El-Naggar, 2007) and changes in water quality ( Saad et al., 2011 , Abou-El-Gheit, et al., 2012.
Biochemical Composition:
Study of the main constituents of the biochemical composition (water, protein, lipid, ash, and carbohydrate
contents) of fish flesh and the effect of pollution (sewage, industrial and agricultural drainage) on these
constituents are very important for determining the nutritive value of fish (Shakweer et al., 1998 and Tayel,
2003). Also these studies used as an indicator of the stress of pollution ( Tayel, 2007). There are many factors
which affect the biochemical composition of fish flesh including; fishing area, type of food organism , water
quality (salinity, pH and temperature), fish size, stage of maturity and pollution (Tayel, 2003). The obtained
values of biochemical composition including water, protein, lipid, ash, and carbohydrate contents are shown in
Table (3).
Water content:
The results of the present study show that the values of water contents are ranged between 78.92 % and
81.30 %. These values agree with that obtained by Kandil (1987) and Tayel (2003 & 2007), who explained the
depletion in water content to the activity of proteolytic enzymes that minimize the water holding capacity of
tissue and the proteolytic activity was markedly variable according to the pH value changes and they also added
that the decrease in the extracellular fluid volume ( because of dehydration of the whole animal, under the stress
of pollution ).
Protein content:
Protein is the basic building nutrient of any growing animal and usual account for (68-85) % of dry matter
of most fish species (Tayel, 2007).The obtained data show that the values of protein content ranged from 79.12
% to 80.13 % . These results agree with that obtained by Mohamed and Gad (2008). The decrease in protein
content may be due to depletion of oxygen ( Massoud et al., 1973 ; Tayel, 2003 ) or increase in salinity (ElEbiary et al., 1997 and Tayel, 2007) .The above mentioned decrease in protein content may be explained as due
to stress of pollution in lake Qarun, which led to liver and gill breakdown (Haggag et al., 1993 and
Rajamanickam and Muthuswamy, 2008), that rise glucose level and consequently decrease of insulin level
causing decrease in muscle protein content (Tayel, 2003 ; Bgum, 2004). The lowest value of protein content
also may be due to consumption of protein for gonadal development (Wassef, 1985 ; Vutukuru, 2005) or may be
due to low temperature and food availability (Khallaf et al., 1993 ; Mohamed and Gad, 2008). However, rise in
the value of protein content can be attributed to the combined effects of high food availability to storing material
prior to spawning ( Bayomy et al., 1993 ; Bgum, 2004).
Depletion in protein showed that it was taken as an alternative source of energy demand that induced by
different pollutants in lake Qarun as previously reported by Vutukuru, (2005).Also,the depletion in tissue
proteins may be due to impaired of protein synthesis (Rajamanickam and Muthuswamy, 2008), their utilization
in cells repair and organization (Mohamed and Gad, 2008 ) and / or the decrease in uptake of amino acids into
the polypeptide chain. The present findings are in agreement with previous reports of decreased level of tissue
protein on exposure to heavy metals (Rajamanickam and Muthuswamy, 2008) and pesticides (Bgum, 2004).
Lipid content:
Lipid plays a very significant rate in toleost fish with respect to reproduction as an important source of
energy (Tayel, 2007). As precursors of steroids and structural components in all reproductive as well as somatic
cell (Harris, 1992 ; Cicik and Engin, 2005). In contrast to mammals, fish prefer to utilize lipid rather than
carbohydrates as their main source of energy as indicated by Black & Skinner (1986) and Desai et al. (2002).
Lipid is the most variable component in fish especially for adult (Love, 1970 ; Mohamed and Gad, 2008). Lipid
is affected by spawning cycle, food availability, seasonal variations and biochemical activity of fish (Bayomy, et
al., 1993).
The lipid values obtained in this study varied from 10.12% to 14.21%. These values are in agreement with
those obtained by Tayel (2007) and Mohamed and Gad (2008). The low values may be due to spermatogenesis
and oogenesis (Phospholipid and lipoproteins) during spawning season (Tayel, 2003 ; Cicik &Engin, 2005).
3865
J. Appl. Sci. Res., 9(6): 3857-3872, 2013
Ash content:
The obtained ash content values were in the range of 3.80% – 8.70% .As a general, it can be noted that an
inverse relationship was exist between ash content and water pollution as the ash content decrease according to
the stress of pollution ( Tayel, 2003&2007 ; Mohamed and Gad, 2008 ).
Carbohydrates content:
The data obtained for carbohydrate content revealed that there was a great fluctuation. The changes in
carbohydrates are not related to sex, season or region and are not affected by pollution as cited by Tayel
(2003&2007) ; the values of carbohydrates in this study ranged from 0.70% to 1.87%.
Table 1: Physicochemical analysis of lake Qarun water during spring, 2011.
AT
WT
EC
PH
Salinity
DO
BOD
COD
Ammonia
Nitrite
Nitrate
Phosphate
Silicate
N
Minimum
Maximum
Mean
Std. Deviation
Std. Error
4
4
4
4
4
4
4
4
4
4
4
4
4
20.9
18.8
24368
6.5
18.5
4.4
0.83
48.5
760
1.36
34.6
0.079
6.85
23.3
20.9
37650
7.4
27.3
7.96
2.98
135.2
1278
2.6
50.2
0.168
9.62
22.175
19.95
31944.5
7.05
23.425
6.115
2.01
96.525
994.25
1.9875
42.275
0.11525
7.7875
1.08128
0.939858
5639.314
0.404145
4.196328
1.469546
0.957671
40.3926
221.4533
0.544388
7.632988
0.039033
1.248556
0.54064
0.469929
2819.657
0.202073
2.098164
0.734773
0.478835
20.1963
110.7267
0.272194
3.816494
0.019517
0,624278
Table 2: Correlation coefficient matrix of some physicochemical parameters in water of lake Qarun during spring , 2011
AT
WT
DO
COD
BOD
Ammonia
pH
Salinity
AT
1.000
WT
0.999**
1.000
DO
-0.679
-0.642
1.000
COD
-0.976*
-0.964*
0.821
1.000
BOD
-0.978*
-0.969*
0.788
0.996**
1.000
Ammonia
-0.340
-0.294
0.914
0.538
0.506
1.000
pH
0.934
0.948
-0.411
-0.838
-0.840
-0.009
1.000
Salinity
-0.970*
-0.957*
0.836
0.999**
0.991**
0.557
1.000
0.830
EC
-0.938
-0.931
0.741
0.959*
0.980*
0.494
0.949
0.775
Nitrite
-0.990*
0.721
0.987*
0.994**
0.408
0.981*
0.994**
0.893
Nitrate
0.154
0.179
0.221
-0.094
-0.186
0.240
0.116
-0.055
Phosphate
0.937
0.925
-0.801
Silicate
0.756
0.732
-0.887
**Correlation is significant at the 0.01 level (2-tailed)
*Correlation is significant at the 0.05 level (2-tailed)
-0.974*
-0.987*
-0.564
0.758
-0.967*
-0.866
-0.876
-0.787
0.476
-0.863
Table 3: Biochemical analysis of the muscles of Tilapia zillii inhabiting lake Qarun
Parameter
Range
Water content %
78.92 – 81.30 %
Protein content %
79.12 – 80.13 %
Lipid content %
10.12 – 14.20 %
Ash content %
3.80 – 8.70 %
Carbohydrates content %
0.76 – 1.87 %
EC
Permissible limits
WHO,(1993)
C˚
C˚
μmohs
μg/l
mg/l
mg/l
mg/l
μg/l
μg/l
μg/l
mg/l
mg/l
25-35
400-1400
7-8
6-14
Up to 6
Up to 10
50-500
None
2500-5000
400-500 Μg/l
1-10
Nitrite
Nitrate
Units
Phosphat
Silicate
1.000
0.970*
1.000
-0.362
0.995**
-0.912
-0.205
0.969*
-0.821
1.000
0.281
1.000
0.253
0.933
Mean SD
80.10 ± 1.682
79.62 ± 0.714
12.16 ± 2.88
6.25 ± 3.46
1.32 ± 0.78
1.000
3866
J. Appl. Sci. Res., 9(6): 3857-3872, 2013
Di
St
2
3
Hn
N
Ft
4
5
Ft
N
Hr
D
6
7
Fig. 2: Liver section of a control Tilapia zillii fish stained with H&E showing: Hepatic artery (HA), Hepatic
porter vein (HPV), Hepatic cells (HC). (X 100)
Figures (3 -7): Liver sections of Tilapia zillii fish obtained from lake Qaroun stained with H&E
Fig. 3: dilation (Di) and stagnant (St) in blood vessel , X 100 .
Fig. 4: fatty degeneration (Ft) around branching blood vessel, X 400.
Fig. 5: hemosidrin (Hn) inside blood vessels, X 400.
Fig. 6: fatty degeneration (Ft)and degeneration (D) between hepatocytes , X 400.
Fig. 7: severe hemorrhage (Hr) inside blood vessel and necrosis (N), X 400.
3867
J. Appl. Sci. Res., 9(6): 3857-3872, 2013
N
Hr
8
9
Hs
Hr
Hn
Hn
10
11
Hr
Hn
D
Hn
N
Hr
12
13
Fig. 8: Spleen section of a control Tilapia zillii fish stained with H&E showing: Red pulp (Rb), White pulp
(Wb) and Hematopiotic tissues (Ht),X 400
Fig. 9-13. Spleen sections of Tilapia zillii fish obtained from Qaroun stained with H&E.
Fig. 9: hemorrhag (Hr) and necrosis (N) in spleenic tissues, X 400.
Fig. 10: hemosidren (Hn) and hemorrhag (Hr) in spleenic tissues, X 400.
Fig. 11: hemolysis (Hs) and hemosidren (Hn) in spleenic tissue, X 400.
Fig. 12: hemorrhag (Hr), hemosidren (Hn) and necrosis (N) in spleenic tissues, X 100.
Fig. 13: degeneration (D), hemorrhag (Hr) and hemosidren (Hn) in spleenic tissues, X 400.
3868
J. Appl. Sci. Res., 9(6): 3857-3872, 2013
Hr
S
D
14
15
Hr
Hn
D
16
17
N
S
Hr
18
19
Fig. 14: Longitudinal section of gill of a control Tilapia zillii fish stained with H&E showing: primary lamellae
(P L) and secondary lamellae (S L), X 400
Figure (15 -19): Gills sections of Tilapia zillii. fish obtained from Qaroun stained with H&E, X 400.
Fig. 15: Hemorrhage (Hr) in primary lamellae and separation (S) in epithelial cells of secondary lamellae.
Fig. 16: hemosidren (Hn) in primary lamellae.
Fig. 17: severe hemorrhag (Hr) in primary lamellae and degeneration (D) in secondary lamellae.
Fig. 18: separation (S) in epithelial cells of secondary lamellae.
Fig. 19: hemorrhag (Hr) in primary lamellae and necrosis (N) in epithelial cells of secondary lamellae.
3869
J. Appl. Sci. Res., 9(6): 3857-3872, 2013
Dr
Ed
M
20
Hd
Hn
N
21
22
Hr
Ft
Hn
E
N
23
24
E
D
N
D
25
26
Fig. 20: Vertical section of skin of a control Tilapia zillii fish stained with H&E showing: epidermal (Ed),
dermal (Dr) and hypodermal (Hd) layers. (X 400)
Fig. 21: Vertical section of muscle of a control Tilapia zillii. fish stained with H&E
(M). (X 100)
Showing : muscle fiber
Fig. 22-26: Vertical Section of skin & muscle of Tilapia zillii. fish obtained from lake Qaroun stained with
H&E. (X 400)
Fig. 22: Necrosis (N) and hemosidrin (Hn) in hypodermal layer .
Fig. 23: Hemorrhage (Hr), necrosis (N) hemosidrin (Hn) in hypodermal layer
Fig. 24: Eadema (E) and fatty degeneration (Ft) between muscle fiber .
Fig. 25: Degeneration (D) and eadema (E) between muscle fiber.
Fig. 26: Balloon necrosis (N) in hypodermal layer and degeneration (D)in muscle fiber
3870
J. Appl. Sci. Res., 9(6): 3857-3872, 2013
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