effect of some heavy metals pollution in lake mariout on

Egypt. J. Comp. Path. & Clinic. Path. Vol. 21 No. 3 (September) 2008; 191 - 201

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Referred byReferred by Prof. Dr. Ismail A. Eissa Professor of Fish Diseases, Fac. Vet. Med.,

Suez Canal University Prof. Dr. Ragb El-Shawarby Professor of Toxicology and Forensic Medi-

cine, Fac. Vet. Med., Banha University

Effect of some heavy metals pollution in Lake Mariout on Oreo-chromis niloticus fish

By Arafa, M. M.* and Ali, A. T.**

* Biochemistry Dept. & ** Fish Disease Dept. Animal Health Res. Inst., Dokki, Giza, Egypt.

SUMMARY

C hemical analysis of water samples from Lake Marriout during sum-mer, represented abnormal high levels of the heavy metals lead, cad-

mium, copper, zinc and iron. The heavy metals concentration was deter-mined in musculature samples of 10 Oreochromis niloticus fish collected from the lake. The influence of the heavy metal pollution in the lake on some musculature biochemical constituents of the fish was also studied. The current study showed significant elevation in the lead, cadmium, cop-per, zinc and iron levels in musculature comparing with reference fish collected from a relatively clean aquaculture. The fish musculature pro-tein, lipid, calcium, phosphorus and the amino acids, methionine, lysine and cystine concentrations were significantly decreased.

INTRODUCTION

L ake Mariout is a salt lake, or rather brackish with an area of

about 250 square km in northern Egypt. It is separated from the Mediterranean Sea by the narrow isthmus on which the city of Alex-andria was built. The lake shore is home to fisheries and saltworks. Some of the marshy areas around the lake have been reclaimed for new buildings as the city grows. Lake Mariout is highly polluted. It receives the drastic human impacts among the Egyptian lakes. It has

been greatly deteriorated from a productive lake to a heavily pol-luted and highly eutrophicated ba-sin (Hamza, 1999). Also Lake Mariout is highly polluted with different heavy metals such as iron, copper and zinc (Saad et al., 1981 and El-Bestawy, 2000).

Metal contamination of the aquatic environment may lead to deleterious effects from localized inputs which may be acutely or chronically toxic to aquatic life within the affected area. Adverse


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effects of heavy metals pollution at concentrations higher than the ac-ceptable limits have been previ-ously reported (GESAMP, 1985; 1988). Heavy metals have long been recognized as serious pollut-ants of the aquatic environment. They cause serious impairment in metabolic, physiological and struc-tural systems of fish when present in high concentrations (Tort et al., 1987). In the aquatic environment, heavy metals in dissolved form are easily taken up by aquatic organ-isms where they are strongly bound with sulfhydril groups of proteins and accumulate in their tissues (Hadson, 1988 and Kar-gyn, 1996).

The accumulation of heavy

metals in the tissues of organisms can result in chronic illness and cause potential damage to the population (Holcombe et al., 1976 and Barlas, 1999). The salts of cadmium, zinc, copper, lead and iron as well as the mercury com-pounds are the most toxic forms of heavy metals. A combination of zinc or cadmium with copper in-creases the toxic effect of copper several times, representing a syner-gistic action (Metelev et al, 1994).

The tissue of greatest concern regarding heavy metal accumula-tion, from human standpoint, is the musculature. Most of the heavy metals bioaccumulate poorly in fish musculature only in the heav-

ily contaminated environment (El-Safy, 1996).

The present work aims to as-

sess the levels of some heavy met-als in Lake Mariout water and its Oreochromis niloticus fish muscu-lature. Also, studying the associ-ated alteration in some biochemi-cal constituents of such muscula-ture.

MATERIAL AND METHODS

1- Water samples and analysis: Lake Mariout was formed of four basins; the lake proper, the fish farm, the southeast and the south-west basins. Our study was under-taken on the south west basin which had the history of highest levels of heavy metal pollution. Ten water samples were collected in sterile glass bottles from the lake and from a relatively clean fish aquaculture as a reference in summer 2008. Each water sample was subjected to chemical analysis for the heavy metals; lead, cad-mium, copper, zinc and iron using atomic absorption spectropho-tometry after Jackson (1973), pH, calcium, phosphorus, magnesium and sulphates according to Fresen-ius et al. (1988). 2- Fish:

Ten Oreochromis niloticus fish of 120-150 grams body weight were collected alive from the same sites of water collection and at the


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same time. The fish were immedi-ately subjected to examination. Musculature specimens were taken from the fish.

3- Preparation of musculature

specimens: Tissue homogenates were

prepared from the musculature specimens according to Upreti et al. (1991).

4- Biochemical analysis of tissue homogenates:

The muscle homogenates were digested according to Cot-tenie (1980). The lead, cadmium, copper, zinc and iron levels in the homogenates were measured ac-cording to Jackson (1973). Protein concentration in muscle tissue ho-mogenate was determined accord-

ing to Torten and Whitaker (2006). Lipid contents in fish mus-culature were determined using the method described by Johann and Lentini (1971). The calcium con-tent was determined according to Reynolds and Linde (1963). Mus-culature phosphorus concentration was measured after James (1995). The amino acids, methionine, ly-sine and cystine concentrations were determined in musculature according to Gehrke et al. (1987).

RESULTS

Water chemistry: Table (1) represents the pH

value and the concentrations of calcium, phosphorus, sulphate, ni-trite, lead, cadmium, copper, zinc and iron in Lake Mariout and ref-erence aquaculture waters.

Table (1) Some chemical properties of Lake Mariout and reference fish aquaculture waters (mean; n = 10)

Parameter Reference aquaculture Lake Mariout pH Calcium (mg/L) Phosphorus (mg/L) Sulphate (mg/L) Nitrite (mg/L) Lead (mg/L) Cadmium (mg/L) Copper (mg/L) Zinc (mg/L) Iron (mg/L)

7.0 24.5 0.3 52.0 0.03 0.2

0.001 0.002 0.03 0.05

7.0 28.6 6.9

100.0 0.05 0.46 0.27 0.09 0.04 0.08


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Heavy metal concentration in musculature:

As shown in table (2), the concentrations of the lead, cad-mium, copper, zinc and iron were

significantly increased in the mus-culature of Lake Mariout Oreo-chromis niloticus fish comparing with those of the O. niloticus of the reference aquaculture.

Table (2) Heavy metals concentration in the musculature of Oreochromis niloticus of Lake Mariout and reference aquaculture (mean ± SE, n=5)

Group Heavy metal (mg/L)

Reference fish Lake Mariout fish

Lead 0.172 ± 0.011 2.774 ± 0.125**

Cadmium 0.047 ± 0.003 0.240 ± 0.015**

Copper 0.012 ± 0.001 0.342 ± 0.021**

Zinc 54.265 ± 2.115 61.185 ± 2.144*

Iron 0.335 ± 0.020 0.413 ± 0.023*

* significant difference by t-student test at p ≤ 0.05. ** significant difference by t-student test at p ≤ 0.001

Musculature biochemical con-stituents:

It is obvious from table (3) that the concentrations of protein, lipid, calcium, phosphorus and the amino acids methionine, lysine and cystine in the musculature of Ore-ocromis niloticus of Lake Mariout

were significantly lower than those of the fish of the reference aqua-culture.


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Table (3) Some biochemical musculature constituents in Oreochromis niloticus of Lake Mariout and reference aquaculture

Group Constituent

Reference fish Lake Mariout fish

Protein (g/100g) 36.40 ± 1.15 30..21 ± 1.06**

Lipid (g/100g) 6.85 ± 0.14 4.72 ± 0.11**

Calcium (mg/100g) 435.15 ± 12.25 390.64 ± 13.50*

Phosphorus (mg/100g) 710.35 ± 20.76 640.58 ± 19.28*

Methionine (g/100g) 0.74 ± 0.02 0.34 ± 0.01**

Lysine (g/100g) 3.60 ± 0.12 2.14 ± 0.11**

Cystine (g/100g) 0.84 ± 0.02 0.21 ± 0.01**

* significant difference by t-student test at p ≤ 0.05. ** significant difference by t-student test at p ≤ 0.001.

DISCUSSION

H eavy metal pollution is re-garded as a severe problem

because it injures the biological functions of the aquatic organisms and their accumulation in fish or-gans and flesh leading to serious healthy hazardous to the consum-ers (Daoud et al., 1999).

Regarding water chemistry, table (1) represents the means of some water parameters of Lake Mariout and reference aquaculture. As presented in the table, the con-centrations of the heavy metals; lead, cadmium, copper, zinc, and iron were drastically elevated (0.46, 0.27, 0.09, 0.04 and 0.08

mg/L respectively). The acceptable limits of lead, cadmium, copper, zinc and iron don’t exceed 0.03, 0.004, 0.006, 0.05 and 0.05 mg/L respectively for the health and nor-mal physiological functions of liv-ing fish according to Swann (1997).

Concerning the heavy metal residues in the Oreochromis niloti-cus fish musculature, the data shown in table (2) revealed signifi-cant increases in lead, cadmium, copper, zinc and iron levels com-paring with the reference fish. The concentrations of lead, cadmium, zinc and iron in the fish muscula-ture (2.774, 0.240, 61.185 and


196

0.413 mg/ L respectively) were higher than the permissible limits of food for consumers which are 0.05, 0.01, 0.5 and 0.3 mg/L (ppm) for lead, cadmium, zinc and iron respectively (Egyptian Organiza-tion for Standardization, 1993) . However the level of copper (0.342 mg/L) in musculature was still within the acceptable limit (< 1 ppm) (Egyptian Organization for Standardization, 1993).

Regarding biochemical stud-ies of musculature, the concentra-tion of protein in Oreochromis niloticus of Lake Mariout was sig-nificantly decreased comparing with the fish of the reference aqua-culture (table 3). The present result is in agreement with that of Sobha, et al (2007) who found that the ex-posure of the freshwater fish Catla catla to cadmium chloride caused fall in musculature protein content. Proteins are highly sensitive to heavy metal poisoning (Jacobs et al., 1977). When an animal is un-der toxic stress, diversification of energy occurs to accomplish the impending energy demands and hence the protein level is depleted (Neff, 1985). So, the decrease of musculature protein concentration in the present study may be due to consuming the protein in energy production under the conditions of heavy metal stress in the Lake Mariout.

The lipid concentration of

Lake Mariout fish musculature was significantly declined against the lipid concentration in reference fish musculature (table 3). De-creased musculature lipid levels were previously recorded in the freshwater fish Catla catla ex-posed to cadmium chloride (Sobha, et al, 2007). The depletion in lipid and protein contents in musculature may be due to their utilization in cell repair and tissue organization with the formation of lipoproteins, which are important cellular constituents of cell mem-branes, and cell organelles present in cytoplasm (Harper, 1983).

Calcium concentration in the current study was significantly de-creased in the musculature of the Mariout O. niloticus fish against the reference fish as shown in table (3). In agreement with the current result, Robert (2003) stated that Fish exposed to high concentration of cadmium quickly develop lack of calcium. Berntssen et al. (2003) stated that dietary cadmium inhibit calcium intake in fish and the cal-cium level in scales tissues de-creased due to remobilization of calcium from these tissues to maintain calcium homeostasis. The decline in the muscular calcium levels in the present study in the Mariout fish may be due to immo-bilization of calcium from muscu-lature to maintain calcium homeo-stasis.

The muscular phosphorus


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level in the present investigation as shown in table (3) was signifi-cantly decreased in the Lake Mari-out fish against reference fish. During heavy metal exposure the demand for energy in the polluted fish is extremely increased (De Boeck et al., 1997). The increased energy demand by fish muscula-ture increases the demand for high phosphorus compounds such as ATP, and creatine phosphate (William et al., 1980). So the de-crease of fish muscular phosphorus in Lake Mariout is possibly due to its consumption in high phospho-rus compounds production to meet the increased demand to energy by the fish under the conditions of pollution.

Lysine and methionine are among the essential amino acids for the teleost fish, while cystine is non essential amino acid. Cystine can be synthesized within the body from methionine. In the current in-vestigation the levels of the amino acids, methionine, lysine and cystine in fish musculature (table, 3) were significantly decreased in Mariout fish comparing with the reference fish. In agreement with our result, Ansari (1986) recorded that the exposure of Channa punc-tatus fish to 1 ppm of copper sul-phate for 84 days leaded to disap-pearance of cystine from the mus-culature. The heavy metals inhibit the intestinal absorption of essen-tial amino acids like methionine

and lysine in fish (Farman-farmaian et al. 1985). So, in the present study, the heavy metal pol-lution perhaps decreased the intes-tinal absorption of amino acids leading to the decline in their fish musculature levels. Heavy metal pollution increase the production of reactive oxygen species and oxi-dative radicals (Winston and Di Giulio, 1991). The amino acids, methionine and cysteine are prone to oxyradicals during oxidative stress (Nandi et al., 2005). The amino acid cystine can be con-verted in the cell to cysteine under demand (Rosenberg, et al., 1967). So the depletion in methionine and cystine in the musculature of Mari-out fish may be due to the attack of amino acids by free oxyradicals produced under the oxidative stress caused by heavy metal pol-lution of the lake water.

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effect of some heavy metals pollution in lake mariout on

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