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Life Science Bulletin, Vol. 7(2) 2010 : 217-222 ISSN : 0973 5453

ACCUMULATION OF HEAVY METALS IN SELECTED TISSUES OF FISH, TILAPIA

MOSSAMBICA COLLECTED FROM THE PENNAR ESTUARY

G. RAVANAIAH and C. V. NARASIMHA MURTHY*

Department of Zoology, Jawahar Bharati Degree and P.G. College, Kavali (A.P.)

ABSTRACT : The present study was initiated to asses the level of heavy metals in different tissues of fish, Tilapia mossambica
collected from the Pennar estuary at Vutukur of Nellore district. Seasonal variation in heavy metal toxicity was recorded
from July,2008 to June,2009 which could be due to aqua and industrial wastewater discharge, into Pennar estuary. In the
present study, accumulation of heavy metals in all the organs of the test fish (Tilapia mossambica) was found to be high
during monsoon season and low during summer season. The low values of pH, dissolved oxygen, salinity, alkalinity, hardness
during monsoon season may enhance the dissolution of particulate form of the metal (carbonates, chloro-complexes and
hydroxides) and increases the amount of ionic species in water corresponding to the increase in level of metals in the organisms.
The higher level of metals in intestine can be attributed to the type of uptake of food of fish i.e., specific mode of nutrition.
Since, the fish, Tilapia mossambica is a detritus feeder and feed either by sucking up the surface layer of the mud or by
grazing on the rock surfaces, leaving to the transfer of mineral particles into the system along with food. In the present study,
the level of certain heavy metals in some tissues exceeded the FAOs (1983) legal limits. The consumption of such fish muscle
may result in lethal effects in secondary or tertiary consumers (human beings) by biomagnification process.

Key words : Heavy metal, Toxicity, Tilapia, Pennar estuary, Industrial pollution.

INTRODUCTION

Estuaries are important sources of food for coastal aquaculture, whereby, they act as natural filters for suspended material and
pollutants and offer effective flood protection for low-lying areas (Mitra et al.,1990 and Reddy & Mohanthy,2003). Human activities
have drastically altered natural concentration of many substances in the estuaries and added numerous new chemicals. Considerable
attention has been focused on the fate of heavy metals and their derivatives in the aquatic environment (Shankar and Manjunatha,1994).
Biomonitoring of heavy metal pollution has been given ever-increasing attention due to the ability of various aquatic organisms to
accumulate these heavy metals. Most of the salts of heavy metals and pesticides produce toxic effect on fish through physical
accumulation (Tilak et al.,2005). Thus, depending on the physico-chemical properties of the toxin, either physical or functional
accumulation may occur or both simultaneously. Heavy metals are capable of accumulating not only in a biological object (fish, phyto
and zooplankton and plants), but also in the sediment. Hence, the present study was initiated to asses the level of heavy metals in
different tissues of fish, Tilapia mossambica collected from the Pennar estuary at Vutukur Nellore and their seasonal variations from
June 2005 to July 2006 to know the extent of heavy metal pollution, probably due to Nellore district industrial wastewater discharge,
by using it as a biological indicator.

MATERIAL AND METHODS

For the bioaccumulation study, fish Tilapia mossambica of 10-12 cm length and 21+2 gm weight were collected, twice per
season. In each sampling six fish of irrespective sex were collected. Different organs viz., gill; intestine, liver, kidney and muscle were
dissected out and separated into different Petri dishes. All the tissues were dried at temperature 120C for 24-48 hrs and ground to fine
powder using a pestle and mortar. Later, known quantities of the fine powder were taken into crucibles and were ashed in a muffle
furnace at 550C for 2 hrs. The ash was digested with concentrated nitric, perchloric and sulfuric acids. Following the acid digestion,
the samples were cooled and dissolved in 1 N nitric acid and made upto 10 ml. Determination of the heavy metals concentration was
carried out by Atomic Absorption Spectrophotometric (AAS-model 302) method as suggested by Topping (1973). A standard mercury
analyzer (ECIL) was used for Hg estimation by cold vapour AAS. The results were expressed as gg-1 dry weight.

RESULTS AND DISCUSSION

The results of the accumulation of selected heavy metals in the selected tissues of fish, Tilapia mossambica that were collected
at Vutukur of Nellore district and their seasonal variations were shown in Tables.1&2.

*Author for correspondence (email : cv.narasimhamurthy@yahoo.co.in)Received 13.03.2010 Accepted 15.06.2010

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218 RAVANAIAH and MURTHY

Table. 1 Showing the accumulation of heavy metals in different tissues of fish, Tilapia mossambica collected from
various seasons at Vutukur of the Pennar estuary during July,2008 to June,2009.

Heavy Tissue Pre-monsoon Monsoon Post-monsoon Summer Annual Fao's

Metal average legal
limits
Cu Gill 32.376 50.0175 17.6985 6.954 26.7615
Intestine 20.558 32.9175 11.2385 6.308 17.7555
Liver 9.2435 13.167 7.3625 4.7785 6.7374 9.5
Kidney 5.054 9.918 3.762 2.0235 5.187
Muscle 7.752 16.0265 5.5005 1.102 7.5905
Seasonal average 14.991 24.4055 9.1105 4.2275
Zn Gill 37.202 55.9835 26.353 19.589 34.7795
Intestine 54.0265 70.642 38.076 29.5735 48.0795
Liver 25.6975 36.347 15.4565 10.108 21.8975 47.5
Kidney 15.504 28.6235 10.4025 8.189 15.7035
Muscle 14.0885 22.0115 3.686 2.1945 10.488
Seasonal average 29.298 42.7215 18.791 13.927
Cd Gill 1.0545 1.501 0.8265 0.2185 0.9025
Intestine 1.729 2.0425 1.1875 0.836 1.444
Liver 1.01175 1.3965 0.3895 BDL 0.6935 0.19
Kidney 0.456 0.646 0.0912 0.0247 0.2945
Muscle 0.494 0.8455 0.1615 0.07885 0.3895
Seasonal average 0.95 1.2825 0.532 0.228
Pb Gill 6.4885 10.089 2.983 0.798 5.0825
Intestine 4.902 8.512 2.717 0.5225 4.161
Liver 1.729 2.90985 0.342 0.06365 1.254 1.9
Kidney 3.648 5.339 0.6365 0.0228 2.4035
Muscle 2.0235 3.5815 0.5035 0.3705 1.6435
Seasonal average 3.7525 6.0895 1.4345 0.3515
Hg Gill 0.817 1.216 0.4085 0.0645 0.627
Intestine 0.532 0.8265 0.2185 0.0304 0.399
Liver 0.114 0.2185 0.0646 BDL 0.1235 0.475
Kidney 0.228 0.4465 0.0931 0.0152 0.1995
Muscle 0.399 0.741 0.1805 0.04085 0.3325
Seasonal average 0.418 0.6935 0.19 0.0304
All the values were expressed in gg-1 fresh weight.

Copper : The accumulation of copper was minimum (2.0235 gg-1) in kidney and the maximum (50.0175 gg-1) in gill (Fig.1). The
accumulation was minimum (1.102 gg-1) during summer season and the maximum (50.0175 gg-1) during monsoon season (Fig.2).
Zinc : The accumulation of zinc was minimum (2.1945 gg-1) in muscle and the maximum (70.642 gg-1) in intestine (Fig.1). The
accumulation was minimum (2.1945 gg-1) during summer season and the maximum (70.642 gg-1) during monsoon season (Fig.2).
Cadmium : The accumulation of cadmium was minimum (0.0247gg-1) in kidney and the maximum (2.0425 gg-1) in intestine
(Fig.1). The accumulation was minimum (0.0247 gg-1) during summer season and the maximum (2.0425 gg-1) during monsoon
season (Fig.2).
Lead : The accumulation of lead was minimum (0.0228 gg-1) in kidney and the maximum (10.089 gg-1) in gill (Fig.1). The
accumulation was minimum (0.0.228 gg-1) during summer season and the maximum (10.089 gg-1) during monsoon season (Fig.2).
Mercury : The accumulation of mercury was minimum (0.0152 gg-1) in kidney and the maximum (1.216 gg-1) in gill (Fig.1). The
accumulation was minimum (0.0152 gg-1) during summer season and the maximum (1.216 gg-1) during monsoon season (Fig.2).

In general, accumulation of all the heavy metals was observed to be minimum during summer and maximum during monsoon
season. Monitoring heavy metals in fish and other food item is concerned with human health protection. In terms of preference, fishes
are the prime seafood of Indians and fishes play a significant role in determining the heavy metal levels of body tissues of Indians who
are consuming seafood. The high level of fluctuation (seasonally) in the uptake of heavy metals in aquatic biota has been attributed to
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EFFECT OF HEAVY METALS ON TISSUES OF TILAPIA 219

Table. 2 Correlation co-efficient (or) for accumulated heavy metals in fish and hydrographical characteristics at
Vutukur of the Pennar estuary.

Heavy metal in fish

Parameters
Cu Zn Cd Pb Hg
Cu in fish 1
Zn in fish 0.996916 1
Cd in fish 0.986752 0.983972 1
Pb in fish 0.995037 0.998052 0.992232 1
Hg in fish 0.998277 0.996742 0.994084 0.998295 1
Rainfall 0.836266 0.850531 0.756054 0.821328 0.814954
Air temperature -0.83076 -0.82082 -0.8034 -0.81318 -0.82202
Surface water temp -0.82688 -0.82084 -0.81162 -0.81773 -0.82316
Turbidity 0.607717 0.628481 0.663858 0.651379 0.633888
Salinity 0.069543 0.081959 0.14867 0.112252 0.099683
pH -0.77183 -0.78128 -0.77897 -0.78618 -0.77921
DO 0.563576 0.572977 0.618053 0.595421 0.586343
BOD -0.7097 -0.69322 -0.75076 -0.7117 -0.7223
COD -0.66758 -0.68635 -0.71786 -0.70724 -0.6915
Total alkalinity -0.63188 -0.64437 -0.67906 -0.66413 -0.65313
Inorg. Nitrate 0.760413 0.772917 0.766928 0.777444 0.76826
Inorg. Nitrite 0.654747 0.666909 0.672521 0.675445 0.665979
Ammonia 0.637896 0.631001 0.675137 0.647514 0.651206
Inorg. PO2 0.642104 0.654755 0.676493 0.669627 0.659069
Sulphate -0.69654 -0.68195 -0.74098 -0.70163 -0.71069
Chloride -0.5087 -0.53788 -0.55961 -0.55822 -0.53473
Fluoride -0.40294 -0.44607 -0.43975 -0.46026 -0.42708
Total hardness -0.65228 -0.66196 -0.72055 -0.68997 -0.68013
Calcium -0.71933 -0.74148 -0.76407 -0.7603 -0.74233
Magnesium -0.65986 -0.66215 -0.71525 -0.68545 -0.6816
Cu in water 0.701064 0.687529 0.596502 0.649896 0.664704
Zn in water 0.810801 0.773279 0.75802 0.756587 0.786814
Cd in water -0.00903 0.057413 -0.06281 0.034806 -0.01171
Pb in water 0.66396 0.64325 0.758986 0.682462 0.693794
Hg in water 0.114136 0.040576 0.090808 0.03406 0.089229
Cu in sediment 0.822931 0.798071 0.75339 0.77461 0.796263
Zn in sediment 0.925996 0.909988 0.882772 0.896718 0.910748
Cd in sediment -0.83074 -0.79257 -0.80717 -0.78719 -0.81663
Pb in sediment 0.481236 0.533431 0.540544 0.556255 0.515458
Hg in sediment -0.58132 -0.62967 -0.65188 -0.65724 -0.61882
60 45

40
50
35
Metal conc. (g/g)

40
Metal conc. (g/g)

30

30 25

20
20
15
10 10

5
0
Gill Intestine Liver tissue Kidney Muscle 0
Pre-monsoon Monsoon Post-monsoon Summer

Copper Zinc Cadmium Lead Mercury

Fig. 1 Showing the annual average concentration of heavy metals Fig. 2 Showing the seasonal average concentration of heavy metal
in the tissues of fish, Tilapia mossambica collected from the in fish, Tilapia mossambica collected from the Penner estuary
Penner estuary at Vutukur during July,2008 to June,2009. at Vutukur during July,2008 to June,2009.
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220 RAVANAIAH and MURTHY

the major interrelated factors by many authors like (i) pollutant delivery to the aquatic environment (Daniel,1990), (ii) the form of the
metal in water (Ayyavoo,1995), (iii) changes in the ambient water quality parameters (temperature, salinity, pH, dissolved oxygen,
nutrients, hardness, turbidity, alkalinity, etc.) (Daniel and Joseph,1991), (iv) organisms physiology, particularly sexual cycle
(reproductive condition) (Zingde et al.,1976), (v) ability to regulate the uptake and elimination of pollutants (excrete) (Babu,1995),
(vi) activity of the animal, species, age, sex weight, stage of life history, body condition and acclimatization of metals (Parimala,1994)
and (vii) other physical factors like nature and concentration of heavy metal, its valence, the association, anions, time of exposure,
volume of water, rate of flow, site of sampling, chelation by organic complexing substances, etc. (Senthilnathan &
Balasubramanian,1997,1998 and Rao et al.,1998).

The absolute concentration of heavy metal depends upon physical and chemical characteristics of the water (Sivakumar et al.,
1988). Changes in ambient water quality parameters (like temperature, salinity, pH, dissolved oxygen, alkalinity, etc.) are interrelated
in changing the water quality and speciation of the metals. This alters the toxicity of the metal and the biological availability of the
metal. In the present study, the higher levels of accumulation of heavy metals in all the organs of the test fish, Tilapia mossambica was
found to be high during monsoon season and low during summer season. Sprague (1971) has stated that the concentration of a
pollutant alone is not a reliable criterion in the accumulation; other physical and chemical factors are also very important. In the
present study, temperature has a minimum impact on the accumulation of heavy metals in the test fish. The probable reason for this
would be that in the tropical regions like India, temperature range at different seasons are not great and the animal was no found to
show much correlation with the temperature fluctuation.

The low values of pH, dissolved oxygen, salinity, alkalinity, hardness during monsoon season may enhance the dissolution of
particulate form of the metal (carbonates, chloro-complexes and hydroxides) and increased the amount of ionic species in water
corresponding the level of metals in the organisms, as observed by Mitra et al. (1994,1995); Muralidhran & Raja (1997) and
Chakravarthy & Vass (1999). These observations are also in corroborate with the findings of the present study, where the heavy metal
accumulation is more during monsoon season that is with low pH, salinity, temperature and high turbidity and dissolved oxygen.
Salinity is to be viewed as an extremely complex variable, affecting not only the physico-chemical characteristics of trace metals (i.e.
the amount of trace metal that remains in solution-ionic form - is a function of solvents) in the marine environment, but also in the
animal tissues (Arul et al.,1990). According to Patil (1998), in hard water, the toxicity decreased in comparison with soft water. High
alkalinity, hardness and pH also appear to play a vital role in regulating the degree of metal toxicity on fish.

The seasonal order of accumulation of metals in fish, Tilapia mossambica collected at Vutukur station were found to be :

Monsoon > Pre monsoon > Post monsoon > Summer.

In the present study, the heavy metals in water were more during monsoon season. Hence, the occurrence of high accumulation
levels of metals in fishes during monsoon season could be attributed to the background levels under the favorable physico-chemical
conditions. The other possible reason for high level of heavy metals in fish tissues during monsoon season might be attributed to the
physiology of the fish, as these fishes shows higher feeding rates between the months of October to December, as the breeding season
or pre spawning period occurs between these months. Babu (1995) has also made similar observation from the fishes of Bay of
Bengal.

On the contrary, during summer, in addition to the reasons quoted against monsoon season, photochemical decomposition of
certain organically bound metals (e.g. Methyl mercury) happens resulted in the evaporation of metals from seawater (Babu,1995). In
general, the accumulation of copper and zinc was more probably due to their essential nature, as they have biological importance as
constituents of metal enzymes (Senthilnathan and Balasubramanian,1997&1998). The studies on the distribution of heavy metals in
different organs of an organism are important to understand not only the extent of environmental pollution, but also the role of these
metal ions in metabolic processes. During the process of intake, some of the metals actually get accumulated in the body of the organs,
because of the affinity between certain metals and certain organ systems (Rao et al.,1998).

It is evident from the present study that the order of accumulation of metals in fish, Tilapia mossambica collected at Vutukur
were found to be :

Zn > Cu > Cd > Pb > Hg

This can be attributed to the point sources of heavy metals to the ambient medium at that station. Similar observations were
also made by Parimala (1994) from Marina Beach, Madras; Babu (1995) from Bay of Bengal and Krishnakumari and Nair (1999a&b)
from Thane and Basin creeks of Bombay.
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EFFECT OF HEAVY METALS ON TISSUES OF TILAPIA 221

In general, marine organisms have greater affinity for cations and Zn2+ and Cu2+ (Parimala,1994). In the present study, the
concentrations of essential elements (Cu & Zn) were found to be more in fish than the non essential elements (Cd, Pb & Hg). The
possible reason for this may be that the role of these essential elements as biological constituents of metal entry (Senthilnathan and
Balasubramaniam,1997&1998). The higher concentration of Cu and Zn likely that endogenous or exogenous ligands or chelating
agents combined with the metal ions to form soluble complexes and are transferred into the cells, as reported by Zingde et al. (1976).
In the present study, even though Vutukur is not rich of certain heavy metals, both in water and sediment, surprisingly, the fishes
collected from it were exhibited an higher levels of the metals. This can be corroborated with the observations of Kumaraguru (1980),
who has explained that the fishes move rapidly from place to place to avoid the contaminated areas and hence, the fishes might move
from highly polluted at Vutukur of Pennar estuary in the present study.
The order of accumulation of metals at Vutukur in different tissues of the test fish, Tilapia mossambica was found to be :
Intestine > Gill > Liver > Kidney > Muscle
According to Nigam et al. (1998), the varying levels of pollutants in different organisms may be due to different rate of
metabolism of pollutants and different tissue lipid contents resulting in different biomagnification in the organisms. Pragatheswaran
(1988) has also stated that the bioaccumulation of one metal in tissues cause changes in Bioconcentration of other metals. The
elimination of other metals is attributed to excess accumulation of particular metal ions (competition for metal binding sites in the
protein molecules-metallothioneins-leading to a redistribution of other metal ions from their original binding sites) and hence, the
variance of metal concentrations in tissues. The various mechanisms by which the metal uptake could occur according to Kumaraguru
(1980) are (i) adsorption of ions at membrane water interfaces, as it might happen in gills, (ii) absorption by active or passive
diffusion of metal ions from water across semi-permeable membranes into body fluids and subsequent distribution to other organs,
(iii) ingestion of particulate and suspended matrices of pelagic and sedimentary origin, (iv) uptake of metals after pre-concentration
in food materials and (v) complexity of metals by coordinate linkages with appropriate organic molecules in the incorporation of
metal ions into physiologically important systems.
In the present study the metals were accumulated at higher levels in gill and intestine. The possible reason for the high level of
metals in gills may be due to the route of entry through active or passive diffusion between the gill membrane and actual medium, as
gills are the active sites of respiration and transport system associated with osmoregulation; as they were closely in contact with water,
since, the layer is supposed to have carbohydrates and other sulphides with possible ion exchange properties; they facilitate a rapid
exchange across the gill membranes. Denton and Breck (1981) have explained the presence of high mercury levels in gills are due to
mucous secreting ability since, it can absorb mercury and other polyvalent cations from seawater. The higher level of metals in
intestine can be attributed to the type of uptake of food of fish i.e., specific mode of nutrition. Since, the fish, Tilapia mossambica is
a detritus feeder and feed either by sucking up the surface layer of the mud or by grazing on the rock surfaces, leaving to the transfer
of mineral particles into the system along with food. The sediments of the present study area are already enriched with trace metals
and as a result, these detritus feeders were exposed to more quantity of metals than the fishes that are pelagic in nature (Zingde et
al.,1976 and Krishnakumari & Nair,1999a&b).
The elevated level of metals in intestine can also be attributed to the feeding habits. Since, the fish, Tilapia mossambica largely
prey on invertebrates like crustaceans, molluscs and zooplankton. Other primary consumers have in gut high metal levels in their
food, it is quite obvious to exhibit elevated metal concentrations in body tissues of fish. Babu (1995) also made similar observations
in the ichthyo-fauna of Bay of Bengal. Generally, kidney as a major route of accumulation, as an excretory organ shows a particularly
high affinity for metals, relative to other organs, thus suggesting a prominent role in the metal metabolism (Tulasi et al.,1992).
Increased accumulation of metals in liver and kidney (Vincet and Ambrose,1994). According to Selvi (1980), more accumulation of
metals in physiologically active sites or organs of fish can be expected. The elevated level of metals in liver may be due to the
enhanced capacity of liver to accumulate more of toxic metals (Kasthuri and Chandran,1997).
The decrease in concentrations of metals could be due to either decreased rate of accumulation of due to the fact that, once the
accumulation had reached a limit, proportionally and totally depending on physiological and environmental conditions, the fish
tissues would try to maintain their level by rejecting the excess metals by regulatory processes (Kumaraguru,1980). In the present
study, the level of certain heavy metals in certain tissues exceeded the FAOs (1983) legal limits. Even, the edible part of fish muscle,
which generally shows low accumulation capacity towards metals also exhibited elevated levels for certain metals. The consumption
of such fish muscle may result in lethal effects in secondary or tertiary consumers (human beings) by biomagnification process. Not
only that, since the pollutants leads to the impairment of organisms, survival of organism is naturally taken to have greater ecological
significance, thus survival of one or even a number of individuals. Further, this may even leads to the so called fish kills that have
greater economic significance.
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222 RAVANAIAH and MURTHY

From the present study, it can be confirmed that the fish, Tilapia mossambica is a biological indicator organism to assess the
level of pollution in the Pennar estuary, Nellore, Andhra Pradesh.

ACKNOWLEDGMENT

The authors are thankful to the Principal, Jawahar Bharati Degree & P.G.College, Kavali for providing the research facilities.

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