6. Ijeefus - Lead Accumulation Profiles in the Soft Tissues of The

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LEAD ACCUMULATION PROFILES IN THE SOFT TISSUES OF THE

MACROGNATHUS PANCALUS DURING SUB LETHAL CHRONIC EXPOSURE, IN

LABORATORY

SUBHAMOY DAS1, SAGNIK MANDAL

2, SUSANTA KUMAR CHAKRABORTTY

3,

RAJKUMAR GUCHHAIT4& AVISHEK DOLAI5

1, 2, 4Department of Zoology, Mahishadal Raj College, Mahishadal, Purba Medinipur, West Bengal, India3Department of Zoology, Vidyasagar University, Paschim Medinipur, West Bengal, India

5Department of Zoology, Calcutta University, West Bengal, India

ABSTRACT

Static bioassay methods were used under laboratory conditions to determine the accumulation of lead byMacrognathous pancalus and determined by Atomic Absorption Spectroscopy. After various time (3, 6, 9, 12, 15, 18, 24,

36 and 42 days) of exposure, to100. 200, 300, 400 and 500 ppm of lead nitrate different tissue of Macrognathous

pancalus (brain, kidney, liver, skin, gill and muscle) accumulated lead was remarkably different. The lead was found to

be accumulated to different levels by the different tissues analyzed by AAS. The order of accumulation was brain > liver

>kidney>gill> muscles>skin. The high level of lead in the gill might not be unconnected with externally bound lead from

the medium rather than internally bound lead to the gill.

KEYWORDS: Bioaccumulation, Heavy Metal, Lead, Sublethal, Liver, Skin, Brain, Kidney, Muscle. Macrognathous

Pancalus Hamilton, 1822, Mystacambilideae

Received: Nov 07, 2015;Accepted: Nov 20, 2015;Published: Nov 30, 2015;Paper Id.: IJEEFUSDEC20156

INTRODUCTION

Heavy metals are natural trace components of the aquatic environment, but their levels have increased

due to industrial, agricultural and mining activities. As a result, aquatic animals are exposed to elevated levels of

heavy metals.The levels of metals in upper members of the food web like fish can reach values many times higher

than those found in aquatic environment or in sediments. Thus contamination in the region is an important issue

regarding the health of the aquatic animals and in turn, health of the freshwater food consumers.

Lead and other trace metals have high affinity for animal tissues where they are concentrated to varying

levels (Rabinaw, P. S. and Dallinger, 1993;Huang, 2003; Martinez et al.,2004). Lead as a metal has physical and

chemical properties that make it extremely useful in industries especially in lead battery production, colored inks

and paint preparation in West Bengal specially in Purba Medinipur District. Lead thus constitutes an important

constituent of wastes discharged from industries to which aquatic animals especially fishes are exposed. The up-

take from the medium continues passively against a concentration gradient since backflow is limited by the excess

high affinity binding sites within the body of the fish (Rainbow and Dallinger, 1993).Heavy Metals are non-

biodegradable and which elevated levels form threats to human health through food chain (Goodwin, et al., 2003;

Rauf et al., 2009). Lead is very toxic are known to present greater hazard when they are both persistent andbioaccumulative (DeForest et al., 2007). The quantity of metal accumulated has been reported to be directly

OriginalArtic

le

International Journal of Environment, Ecology,

Family and Urban Studies (IJEEFUS)

ISSN(P): 2250-0065; ISSN(E): 2321-0109

Vol. 5, Issue 6, Dec 2015, 35-50

TJPRC Pvt. Ltd.


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36 Subhamoy Das, Sagnik Mandal, Susanta Kumar Chakrabortty, Rajkumar Guchhait, Avishek Dolai

Impact Factor (JCC): 3.0965 NAAS Rating: 3.63

related to the concentration to which the organisms are exposed and the period of exposure (Allen and Masters, 1985;

Daramola and Oladimeji, 1989; Otitoloju, 2001: Kamaruzzaman et al., 2010). Metals are also preferentially accumulated

by different organs of the body (Bilgrami et al., 1996; Vinodhini and Narayanan, 2008: Rauf et al., 2009). The positive

correlation between the concentration of metals in the aquatic environment and the tissue of the fish (Daramola andOladimeji, 1989: Bu-Olayan and Thomas, 2008) often pose serious health problems to fish consumers especially man.

Oyewo(1998) identified lead as one of the prominent heavy metals in industrial effluents discharged into the Lagos Lagoon

and also featured prominently in the water and sediment samples from effluents receiving water from storm drains.

Consumption of fish with high amount of lead is a major route of human exposure to lead when contaminated fish are

consumed (Abdul-Kashan and Singh, 1999; Dougherty et al., 2000; Nnaji et al.; 2007). Lead is known to be accumulated

in different organs of fish including the bone, gills, kidneys, liver and scales (Dallas and Day, 1993: Javid et al., 2007). In

this study Macrognathous pancalus or pancal were exposed to different sub lethal concentrations of lead nitrate solution

under static laboratory conditions. Macrognathous pancalus or pancal fish is commercially important fresh water benthic

fast moving mud dwelling fish species in wet land of west Bengal and surrounding.. The Macrognathous pancalus is ancarnivore and usually inshore, entering pond, canal, wetland, jheel, river and lagoons. Macrognathous pancalus is

considered to be Least Concern since it is very widely distributed and with limited threat. Macrognathous pancalus is

widely distributed throughout India, Pakistan, Nepal, and Bangladesh. Present in most of the Ganges drainage, but scarcer

in numbers in Nepal. Macrognathous pancalus occurs in running and stagnant waters. Found in fresh and brackish waters

and deltas of large rivers, common in ponds and slow flowing rivers with vegetation in plains. Inhabits still waters with silt

or mud substrate. Believed to be common in rice paddy fields. Nocturnal feeder, on insects and worms.in lowland habitats

and at moderate elevation in all the larger river systems of the Indian subcontinent. Macrognathous pancalus occurs in

lowland habitats and at moderate elevation in all the larger river systems of the Indian subcontinent. It attains a length of

38 cm and is considered a wholesome food fish.

Although there are few reports regarding the lead or any type of heavy metal accumulation in the fish of West

Bengal or India context but there are no report regarding the benthic fish who exposed maximum level of heavy metal due

to their benthic habit. So our study is the pioneer and wonderful effort regarding the benthic species, M. pancalus, specially

in the ecological condition of West Bengal.

Heavy metals have long been recognized as serious pollutants of the aquatic environment. They cause serious

impairment in metabolic, physiological and structural systems when present in high concentrations in the milieu (Tort et al

1987). Heavy metals may affect organisms directly by cumulating in their body or indirectly by transferring to the next

trophic level of the food chain. One of the most serious results of their persistence is biological amplification through the

food chain (Unlu 1993). In the aquatic environment, heavy metals in dissolved form are easily taken up by aquatic

organisms where they are strongly bound with sulfhydril groups of proteins and accumulate in their tissues (Hadson 1983,

Kargn 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, Barlas 1999). The 96-h LC50 tests are conducted to measure.

The 96-h LC50 tests are conducted to measure the susceptibility and survival potential of organisms to particular toxic

substances such as heavy metals. Higher LC50 values are less toxic because greater concentrations are required to produce

50% mortality in organisms (Eaton1995). The heavy metals that are toxic to many organisms at very low concentrations

and are never beneficial to living beings are mercury, cadmium and lead (Hilmy et al 1985). Fish absorb dissolved oravailable metals and can therefore serve as a reliable indication of metal pollution in an aquatic ecosystem (Hilmy er


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Lead Accumulation Profiles in the Soft tissues of the Macrognathus 37

Pancalus during Sub Lethal Chronic Exposure, in Laboratory


al1985). Tench (Tinca tinca) is considered a good test organism for heavy metal contamination because of its feeding

behavior and bottom feeding habits (Guerrin et al 1990) in foreign country. In our country heavy metal accumulation study

were performed on several fish but there were no standard or model fish so far. In the present study we select such a fish

which is very hardy, tolerant, carnivorous, and very important is the pelagic- benthic in habit The present study was carriedout to observe the rate of accumulation of heavy metal Pb in different tissue or organ in the body of Macrognathus

pancalus.

MATERIALS AND METHODS

Fish Macrognathous pancalus were collected from different sites at Uluberia, Bagnan, Mahishasdal, Haldia and

Nayachar Island of West Bengal (Figure 1) near the Mahishadal Raj College with cast nets and from different fisherman.

Afterward the fishes were transferred to the experimental laboratory of the Department of Zoology, Mahishadal Raj

College,Mahishadal, Purba Medinipur. The Macrognathous pancalus fingerlings used in this study weighs between 9.60g

and 14.40g (mean weight = 11.45g).They were brought to the laboratory in polythene bags. In the laboratory, they wereacclimated in aquaria using tap water maintained at 26.0C for few week.

During acclimation, they were fed once a day at 5 percent body weight using pelleted fish feed obtained from the

local market. The water in aquaria was reconstituted every 48 hours during acclimation. The fish were stocked in 5- l

capacity aquaria for 3 weeks to acclimatize to laboratory conditions. The water temperature, dissolved oxygen, pH and

electric conductivity were measured regularly in the laboratory; however, the other physico-chemical parameters were

measured at the Sea Explorar, Kolkata.The fish were fed with dry Tubifex and living zooplankton during the experiment.

Every 2 days the unconsumed food and fecal matter containing water were changed regularly and a proper hygienic

condition were maintained. To investigate the effects of heavy metals already accumulated in the body of Macrognathous

pancalus of the heavy metals lead, the experiments were designed accordingly. For determination of the 96-h LC50 (lethal

concentration) values, 6 concentrations each of lead sulphate E MARK were used.

A group of 8 fish was used for each concentration of each heavy metal concentration. Separate groups of 8 fish

each served as controls for heavy metal- lead. All experiments were run for 96 h and the concentration of each heavy metal

that caused 50% mortality in fish was named the LC50 value of the heavy metal lead. The mortality was calculated as a

percentage once every 24 h. The LC50 values for 96 h were 600.0 ppm for lead sulphate. The LC50 values were found

(11,12) and the true concentration of lead was obtained (13).

The physicochemical parameters of the water were determined before use in the experiments using standardmethods (APHA, 1995). A stock solution (1,000mgl-1) of reagent grade lead nitrate was prepared using distilled water

(Reish and Oshida, 1986). Five (5) sub lethal concentration (100, 200, 300, 400 and 500 mgl-1) of Pb sulphate were used

base on the 96 hours LC50 values. The LC50 represents the concentration of at which 50% of the test fish died after

96hours of exposure. A control solution was prepared without lead.

Fifteen (15) adult of M. panchalus were randomly loaded into 100 liter glass aquaria containing the different

concentration of lead sulphate. The fish were the fed daily a quantity of pelleted food equivalent to five percent of their

body weight and water in the aquaria was reconstituted every 7 days for 42 days.

Fish samples were taken after 3, 6, 9, 12, 15, 18, 24, 30, 36 and 42 days (after this duration all the fish irrespectiveto dose not survive at all.) from the different concentrations and the control group. The organs were isolated and weighed


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separately. Main organ were dissected were liver, gill, kidney, muscle, skin and, brain. One (1) gram (mean) of each organ

samples from the all different concentrations taken during each sampling period were wet-ashes using 10ml freshly

prepared nitric acid. Resulting solutions were gentle boiled to 2 - 5ml and allowed to cool down. Each was then separately

transferred into a 25ml volumetric flask and filtered into a clean glass bottle for analysis. The solutions were analyzed forlead using Atomic Absorption Spectrophotometer. The lead residues in each sample were calculated based on FAO/SIDA

(1983) methods.

Sampling

A map of Nayachar showing the sampling sites is presented in Figure 1 below. Three series of independent

sampling coded named were characteristically chosen to purposively represent the major industrial conditions at the

Nayachar. The sampling was done in February, 2015. This period was generally characterized by very dry and cold.

During the dry season, temperatures were usually very low (over 170C during the day) coupled with low humidity and

intermittent rainfall. The converse is true for the wet season. The mean rainfall during this period ranges from 850 to 1000mm which occurs in the months of February April 2015. The M. pankalas samples were caught by trap anchored using

locally made hoop-fine traps and cast nets at three different locations in the water body of Nayachar from Feb to May

2015. The physicochemical parameters of the water body have recently been done (Pelig-Ba, 2011). The lengths and eights

of each samples were measured (Table 1) and recorded before been frozen in polyethylene bags for transportation to the

laboratory for chemical analysis.

Sample Preparation

All samples were thawed and then thoroughly washed with distilled water followed by double distilled deionized

water at the Centre. A clean high quality rush free and corrosion resistant stainless steel knife was used to cut about 2 g ofmuscle tissue from each sample along the lateral line. Also, the operculum of each fish from each sampling series was

carefully opened and the gills extracted. Again, the livers of each fish sample were obtained after appropriate dissections.

The parts of liver, muscle, skin, brain and gills of all the M. pancalus samples which were carefully categorize per tissues

per sampling series per strata were pooled for further preparation and analysis. The samples were then separately

pulverized using a laboratory mortar and pestle manually. The pulverized samples were finally milled in a Retsch RS 100

model vibratory disc mill and stored at -20 C in closed polyethylene bottles with screw caps until analysis.

Chemical Analysis

Quadruplicates (about 200 mg) of each categorized pulverized sample were accurately weighed into acid cleanpolyethylene foils, carefully wrapped with forceps, capped and then heat sealed. For AAS analysis, digestion beakers, test

tubes and volumetric glassware were first cleaned by a procedure described by Appiah et al. (2012). Four replicates of each

category of pulverized samples were again accurately weighed (approximately 0.5 g) and directly dissolved into Teflon

vessels containing a (6:1 v/v) mixture of HNO3 and HClO4. These vessels were then swirled gradually to form a

homogeneous mixture, fitted into an ETHOS 900 microwave digester and digested for 25 minutes. After appropriate

cooling, each digested solution was transferred into a measuring cylinder and diluted to 20 ml using de-ionised water.

These solutions were then analyzed using a Atomic Calibrations, precision together with accuracy of this system has

previously been done by the authority.


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RESULTS

The morphomertic measurement of the fish of different locality which were collected and used for the

bioaccumulation experiment were presented in Table 1. This result shows that the specimen were mainly collected

from three sites ie, Nayachar of Purba Medinipur, Geonkhali of Purba Medinipur and Uluberia river side canal if

River Ganga. However the site of collection of the fish, in the experimental purpose the average size of the fish of

three sites were11.25, 10.99cm and 10.21cm respectively. The mean weight of the used fish was 8.7g.98,

8.9g.98, 8.23, 1.33 g. The experiment was conducted in the month of March, April and May of 2015. The

moisture content of the above fish was also examined. The average moisture % were 81- 84 2.09.

The physicochemical; parameters (average of three sites) were presented in Table 2. Do2, alkalinity, salinity,

color, hardness, TDS, nitrate, sulphate, cupper, and zinc are determined by following APHA.

The Figure 1, 2, 3, 4, 5and 6 represents the result of the accumulation of lead against different doses(100, 200,

300, 400, 500 ppm) in different time of exposure period (3, 6, 9, 12, 15, 18, 24, 30, 36 and 42 day) were presented in brain,

liver, gill, muscle, kidney and skin respectively.

Table, 3, 4, 5, 6, 7and 8 also represents the data of Bioaccumulation of lead in different tissue (brain, liver, gill,

kidney, skin and muscle)of fish M. pancalus against different time(3, 6, 9, 12, 15, 18, 21, 24, 30, 36, 42 days) of exposure

in 100ppm, 200ppm, 300 ppm, 400 ppm, and 500ppm lead nitrate exposure respectively.

Table 1: Morpho Metric Measurement of the Fish Macrognathous

Pankalus,(At Different Sites) Used for the Bioaccumulation Experiment

Site

Mean Length in cm

(aver, range, SE)

Mean Weight in g

(aver, range, SE) Month

Moisture Content

in % (aver, range,SE )

Nayachar 11.25 (9.1-13.01,1.22 8.7, 7.23-11.8,.98 15-Mar 83,224.23Geonkhali 10.99(10.22-12.55, 1.89) 8.9, 8.23-11.69, 1.75 29-Mar 81.983.41Ulubaria 10.21 (8.45-12.34,) 8.23, 7.29-9.87, 1.33 17-Apr 84.342.98

Table 2: Physico Chemical Parameters of the Water, Habitat of the

M. Pancalus, used for Bioassay and Bioaccumulation Experiments

Parameters Range Mean

Colour TCU 59.00 121.60 96 12.67Temperature 0C 24.6 27.50 23.40 1.21pH 6.5 7.4 6.9 064

Salinity 3.36 6.40 3.50 1.51DissolvedOxygen (ppm)

5.96 6.21 5.7 0.42

Settleable solids 0.39 1.52 0.62 0.15Alkalinity (ppm) 24.36 38.54 33.64 2.05Total hardness asCaCO3 (ppm)

62.25 90.60 72.56 3.90

BiologicalOxygen Demand(BOD) (ppm)

17.65 23.22 19.98 + 3.48

Chloride (ppm) 5.60 8.2 6.42 1.321Sulphate (ppm) 0.00 1.20 0.06 0.044Nitrate (ppm) 2.85 4.96 4.05 0.47

Phosphate (ppm) 0.02 0.68 0.24 0.19Ammonia (ppm) 0.01 0.05 0.01 0.025


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Table 2: Contd.,

Copper (ppm) 0.01 0.03 0.01 0.081Lead (ppm) 0.20 0.41 0.36 0.24Iron (ppm) 0.50 1.64 0.76 0.64

Zinc (ppm) 0.01 0.030.013 0.011

Chromium (ppm) 0.03 0.060.031 0.017

Manganese (ppm) 0.01 0.19 0.03 0.017

Figure 1: Accumulation of Pb in Brain of Macrognathous Pancalus when

Exposed in Various Doses (ppm) and in Various Time (Day)

Figure 2: Accumulation of Pb in Liver of Macrognathus Sp when


Figure 3: Accumulation of pHin Gill of Macrognathous Pancalus when



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Figure 4: Accumulation of Pb in Muscle of Macrognathus Sp when


Figure 5: Accumulation of Pb in Kidney of Macrognathous Pancalus when


Figure 6: Accumulation of Pb in Skin of Macrognathous Pancalus when


Table 3: Bioaccumulation of Lead in Different Tissue of Fish M. Pancalus

Against Different Time of Exposure in 100ppm Lead Nitrate

Time of

Exposure

(Day)

Organ

Brain Liver Gill Muscle Skin Kidney

3 0.06 0.15 0.02 0.22 0.13 0.216 1.2 0.34 0.2 0.23 0.2 0.339 2.5 0.55 1.1 0.25 0.19 0.48

12 4.9 1.5 0.8 0.26 0.199 0.6515 4.6 1.75 0.8 0.29 0.22 0.818 4.9 1.9 0.9 0.32 0.29 1.124 5.56 2.75 1.1 0.46 0.44 1.330 8.55 3.5 1.1 0.52 0.49 1.5

36 9.67 4 1.15 0.75 0.63 2.242 12.9 4.6 1.54 0.82 0.68 2.9


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Exposure

Time

(Day)

Organ

Brain Liver Gill Muscle Skin Kidney3 1.7 0.3 0.2 0.3 0.25 0.19

6 1.77 0.35 2.1 0.32 0.25 2.11

9 2.5 1.65 2.24 0.31 0.25 2.19

12 5.7 1.6 2.5 0.33 0.29 2.35

15 6.3 1.75 2.75 0.35 0.38 2.3

18 6.67 2.6 2.75 0.37 0.37 2.7

24 8.55 3 2.75 0.56 0.49 2.9

30 8.6 3.5 3.75 0.65 NA NA

Table 5: Bioaccumulation of Lead in Different Tissue of Fish M. PancalusAgainst Different Time of Exposure in 300ppm Lead Nitrate

Exposure

Time

(day)

Organ


3 1.1 0.3 0.2 0.31 0.255 0.166 1.93 0.4 2.5 0.42 0.3 0.8669 4.8 0.96 2.55 0.43 0.32 1.312 7.5 1.6 2.55 0.44 0.38 1.7315 7.6 2.4 2.55 0.5 0.44 2.1618 9.8 2.6 4.2 0.5 0.56 2.6



Exposure

Time

(day)

Organ


3 1.4 0.35 0.42 0.79 0.265 0.126 1.4 0.9 0.63 1.06 0.34 .379 5.3 1.6 NA NA NA NA

12 7.56 1.6 NA NA NA NA15 9.6 2.3 NA NA NA NA18 11.9 2.9 NA NA NA NA



Exposure

Time

(day)

Organ


3 1.4 0.02 0.83 0.46 0.27 0.1

6 1.9 0.9 7.8 4.2 NA NA

DISCUSSIONS

Lead is toxic to humans, with the most deleterious effects on the hemopoietic, nervous, reproductive systems and

the urinary tract. The Joint FAO/WHO(2004) Expert Committee on Food Additives establishes a provisional tolerable


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weekly intake (PTWI) for lead as 0.3 mg.kg1 body weight. European Community (No 1881/2006) and Bulgarian Food

Codex (No 31/2006) set maximum permitted level for Pb in fish of 0.4 mg.kg1w.w. In present day, the higher

concentration of lead was measured in gills in various fishes from various part, whereas the edible tissue from this location

presented 0.07 mg kg1 w.w. Various reported for fish species from the middle Black Sea lead levels in the range of 0.220.85 mg kg-1ww. Uluozlu et al. (2007) were investigated lead contents of fish species from Black and Aegean seas and

were found values in range of 0.330.93 mg kg1 for edible fish tissue. In current research, Bat et al. (2012) presented

more higher concentrations for Pb (0.023 mg.kg1 ww) for golden mullet from Sinop region (Turkey Black Sea coast). In

general, analyzed species showed significantly lower Pb concentration in comparison to species from middle Black Sea

and Aegean Sea. Highest Pb levels were obtained in gills in grey mullet from Varna Lake.

The bio accuumulation concentrations of lead, a non-essential heavy metals were determined, by exposing the fish

in 5 sublethal doses (100 pm to 500 ppm) in different tissues of extensively consumed inland fish, Macrognathous

pancalus, found in West Bengal, at different strata by AAS. The brain tissues had the highest of the metals with the skin

tissues recording the lowest. It is therefore recommended that the brain and liver tissues should not be consumed. As the

Macrognathous is very small fish, average weight 10 g, it is impractical to remove the liver and kidney. Specially removal

of kidney is problem some. During dressing of fish it is strictly recommended that the head must be totally removed as in

brain maximum amount of lead was accumulated. As we remove the head totally the gill will also be eliminated.

It is fastidiously noting that, irrespective of the exposure dose and exposure time, the brain tissue had high metal

accumulation than the gills, skin, kidney and muscles. The brain is followed by liver in all the cases irrespective to

exposure time in lead containing environment. It is due to the fact that all entries in the liver row had the highest values for

each metal in all sample. Thus, the dominant accumulating tissues were the liver. One plausible explanation for the high

accumulation potential of the liver is a result of the activity of metallothioneins, metal-binding proteins, which play an

important role in metal regulation and detoxification of nonessential metals (Roesijadi, 1992). Thus, the liver is considered

a good biomonitor of water pollution with metals since their concentrations accumulated in this organ are often

proportional to those present in the environment (Mastan. 2014). The muscle tissues recorded the lower accumulation for

lead. We therefore suggest that, the liver tissues should be removed before using the fish in the preparation of meals as a

safety measure to drastically reduce the level of human exposure to this metals.

The result of Bioaccumulation of lead in different tissue of fish M. pancalus against different time of exposure in

100ppm lead nitrate was presented. In 100 ppm dose the culture survive maximum 42 days(6 weeks).When we considered

the bioaccumd in different organ against the dose 100ppn in tap water at room temperature (27+_ 4) providing dry food inthe laboratory against different exposure time the result varies significantly. At 3 days exposure gill accumulate minimum

(0.02ppm), followed by brain (.06), skin (.13), liver (.15), muscle (.22 and in kidney (0.21ppm). It indicates that immediate

exposure in the toxic substance kidney try to detoxify the lead and so its amount is maximum as in kidney drug detoxifying

mechanism is well established.. In case if kidney the amount is not increased dramatically as the exposure period

increases14 times (42 days), the accumulation increases only 10 times (2.9ppm). But in case of brain the accumulation is

directly proportionate with the exposure time. In the 42th day the amount become 12.9 ppm which is 200 times more than

the 3 days. So the result indicates that at 100 ppm dose brain accumulates maximum amount. The accumulation at 42 days

is 12.9, 4.6, 2.9, 1.54, 0.82 and 0.68 in case of brain, liver, gill, kidney, muscle and skin respectively. The result shows that

in 100 ppm dose as the exposure time increase the brain accumulate more and more but the skin accumulation not


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increased in greater speed. So skin consumption is not so much dangerous as brain for long term lower sub lethal dose. It

also interesting to note that in case of accumulation in brain the 12 to 18 days the amount not varied in large amount. But

when exposure time increased from 36 to 42 days the accumulated amount of lead increase very rapidly (9.6 to 12.9). This

indicates that just before death the rate of the accumulation increased very rapid.The results shows that when toxic material lead nitrate increased twice i.e., 200ppm the result alter a little bit. At

3rdday accumulation was maximum in brain (1.7ppm) like dose 100ppm. Brain is followed by liver and muscle, skin gill

and kidney respectively. The least was gill surprisingly. But in other previous report showed that the gill accumulates the

maximum. We think that there may by 2 reason Firstly, the gill not accumulates but it may externally adsorb. Secondly the

gill and opercular action of the particularly the M. pancalus is very high the fish is benthic in habit. So the gill and opercula

movement ant gill adsorption is much reduced, unlike to other fish. As the exposure period increases the accumulation

increased 8.6(brain), followed by 3.75ppm (gill), 3.5ppm (liver) and0.65ppm (muscle). The result of other two tissues was

not taken as situation was not permitted at 30th days. Although accumulation increased 6 times (1.7 ppm and 8.6ppm)

within 3rdand 30thdays but in case of liver the value increased dramatically that is from 0.3ppm to 3.5ppm that is more or

less 12 times. And gill increased19 folds (from 0.2ppm to 3.75ppm). The muscle increased only two fold within 30 days

(from.ppm to 0.65ppm). Accumulation in skin is less than two fold (from.25 to.49 and in case of kidney bioaccumulation is

least more or less no changes in respect to other organs(0.19 to 2.9ppm)

Result reflects the bioaccumulation of lead in different tissue of fish M. pancalus against different time of

exposure in 300ppm lead nitrate. The bioaccumulation test were continued maximum 18 days because the fish survived

maximum for a period of 20 days. The result shows that at 3 rdday in brain accumulation wer3e maximum (1.1ppm) and at

18th day 9.8ppm, whereas least in kidney(.16ppm). Brain increased 9 times, liver 8 folds, gill 21 folds, muscle1.5 folds and

skin 2 folds and in kidney 1.5 folds. This result shows that the kidney and skin and muscle the lead accumulation was least

at 300ppm also and remain restrict at 1.5 to two fold. But in gill the accumulation increased 21 fold although the maximum

accumulations were observed in brain. When we compared the comparative account muscle is the organ where

accumulation is least (.5ppm) whereas in kidney and liver its 5 times(2.5ppm) and in gill 9 times of the muscle and in brain

20 times of the muscle. It shows that the accumulation in brain is maximum followed by gill. If we cautiously discard the

gill and head during food preparation the biomagnifications through the tropic level will be very minimum. As the dose

increase from 100 ppm,200 ppm, 300 ppm and 400 ppm the accumulation in liver, skin, kidney, brain, gill and muscle

were 1.9, 2.6, 2.62.9 ppm ; 0.29, 0.37, 0.56 ppm; 1.1, 2.7, 2.6ppm; 4.9, 6.7, 9.8, 11.9ppm ;.9, 2.7, 4.2ppm; and 0.32, 0.31,

0.5ppm respectively.

The bioaccumulation of lead in different tissue of fish M. pancalus against different time of exposure in 400ppm

lead nitrate. The main difference between 400ppm and other doses was in this dose maximum accumulation at 6 days was

in brain(1.4ppm) but in 2nd position the organ was not liver (.9ppm) but muscle(1.06ppm).At the dose 400ppm muscle

absorbed more than gill, skin, kidney and liver. So when the fish M. pancalas was exposed in more toxic concentration of

lead nitrate accumulation in muscle increased. So the fish exposed in higher lead concentration is more dangerous than

lower as maximum accumulation occur in brain and followed by muscle.

In case of 4ooppm dose the result obtained is not complete in all organs. Brain increased from 1.4ppm to 11.9ppm

at 18 days whereas liver from.35 to 2.9ppm. So in brain and liver the accumulation rate increase 9 folds and 8.5 fold

respectively. But in brain accumulation is 4time more at both 3rdday and18thday exposure. So the rate of accumulation of


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these two organ brain and liver is same. But as the detoxifition mechanism was is liver the amount of accumulation is very

less in compare to brain. At 6 thday the amount of accumulation was 1.4,.9,.63, 1.06, 0.34 and.37 in brain, liver, muscle,

skin and kidney respectively.

The results of Bioaccumulation of lead in different tissue of fish M. pancalus against different time of exposure in500ppm lead nitrate are presented. As LC 50 dose of 96 hour of the lead nitrate is 650ppm, the experiment of 5ooppm dose

was continued only 8dayd. So the data of 9 days of the 500ppm experiment was tot taken. The result of maximum 6 thday

only obtained. Brain accumulates 1.9ppm, liver.9ppm, muscle4.2ppm and gill 7.8ppm at 6thday. The result shows that at

higher dose but little exposure time the accumulation was higher in gill and followed by muscle. The brain accumulation

was not so high. It in shimmed that in gill the lead was not absorbed but adsorbed. But when we considered the

accumulation at low dose for long time the accumulation in muscle is less, in gill absorption is also less but in brain is very

high. In higher dose but low exposure time the accumulation sequence is gill > muscle >brain >skin > kidney. But in case

of long-term and lower dose the accumulation sequence is brain>liver >kidney >gill >muscle>skin.

Results reflects that in 100 ppm dose the culture survive maximum 42 days (6 weeks).When we considered the

bioaccumulation of lead in different organ against the dose 100ppn in tap water at room temperature (27+_ 4) providing

dry food in the laboratory against different exposure time the result varies significantly. At 3 days exposure gill accumulate

minimum (0.02ppm), followed by brain (.06), skin (.13), liver (.15), muscle (.22 and in kidney (0.21ppm). It indicates that

immediate exposure in the toxic substance kidney try to detoxify the lead and so its amount is maximum as in kidney drug

detoxifying mechanism is well established.. In case if kidney the amount is not increased dramatically as the exposure

period increases14 times (42 days), the accumulation increases only 10 times (2.9ppm). But in case of brain the

accumulation is directly proportionate with the exposure time. In the 42th day the amount become 12.9 ppm which is 200

times more than the 3 days. So the result indicates that at 100 ppm dose brain accumulates maximum amount. The

accumulation at 42 days is 12.9, 4.6, 2.9, 1.54, 0.82 and 0.68 in case of brain, liver, gill, kidney, muscle and skin

respectively. The result shows that in 100 ppm dose as the exposure time increase the brain accumulate more and more but

the skin accumulation not increased in greater speed. So skin consumption is not so much dangerous as brain for long term

lower sub lethal dose. It also interesting to note that in case of accumulation in brain the 12 to 18 days the amount not

varied in large amount. But when exposure time increased from 36 to 42 days the accumulated amount of lead increase

very rapidly (9.6 to 12.9). This indicates that just before death the rate of the accumulation increased very rapid.

The accumulation of heavy metals in aquatic biota has become a major problem. This is because most humans

consume fishes from these polluted water bodies. Pollutants are absorbed and are carried in the bloodstream to the liver fortransformation. Pollutants transformed in the liver may be stored there or transported to excretory organs such as gills,

kidneys for elimination or rather stored in fat which is an extra hepatic tissue [Dimari 2008]. Metals generally enter the

aquatic environment through atmospheric deposition, erosion of geological milieu or due to anthropogenic activities caused

by industrial effluents, domestic sewage and mining wastes.

The term heavy metals refers to any metallic element that has a relatively high density and is toxic or poisonous

even at low concentration [Hutton, 1086]. Heavy metals are generally a collective term which applies to the group of

metals and metalloids with atomic density greater than 4g/cm or 5 times or greater than water [Garbarino et al 1995,].

There are three main routes through which heavy metals enter into the environment. These routes include disposal of metal

enriched sewage sludge and sewage effluents into water bodies, occur as by-products from metal mining processes and


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deposition of atmospheric particulates. These metals are transported through water bodies as either dissolved metals in

water and sometimes as an integral part of suspended sediments. The dissolved heavy metals in water have the greatest

potential of causing the most deleterious effects. The metal contaminants in aquatic systems usually remain either in

soluble or suspension form and finally tend to settle down to the bottom or are taken up by organisms. The progressive andirreversible accumulation of these metals in various organs of aquatic creatures ultimately leads to metal-related diseases

because of their toxicity and thereby endangering the aquatic biota. They then get stored in bed sediments of water bodies

or seep into the underground water, thus causing the water sources to be contaminated. Water bodies such as rivers,

lagoons get contaminated with these heavy metals through human activities like mining, manufacturing, agriculture among

others. These activities introduce wastes containing some of these heavy metals into water bodies. These metals dissolve

and move down stream into lower reaches of the water bodies. Some also settle into the sediments of the water bodies.

The lead burden in C. gariepinus exposed to different concentrations of lead nitrate solution was found to be a

function of concentration and duration of exposure. The Copper residues in Clarias angularis expose to 0.027, 0.055 and

0.11mmg cul-1 were 15.7, 21.8 and 31.17 mg g-1 respectively after eight weeks exposure (Daramora and Oladimaji,

1989). Analysis of fish metal burden in O. niloticus also showed many fold increases above the existing one in the medium

(Onwumere and Oladimeji 1990; Bu-Olayan and Thomas, 2008; Edem et al. 2009).

Different organs in the body are known to accumulate a particular metal to a high level while others do not

accumulate the metal through present in the medium (Javid et al., 2007; Al-Kahtam, 2009). Where accumulation takes

place, kidney, liver and spleen accumulate more metals compared with muscle tissues in metal polluted media (Canli et al.,

1998; Vinodhini and Narayanan, 2008).

Preferential accumulation of metals in the liver, kidney and gills has also been reported by Panigraphi and Misra

(1978; Jantateemes et al., 1999; Raoud and Al-Dahshan, 2010). Muscles are known to show lower metal and

concentrations than other organs (Gachter and Geiger 1979; Falusi and Olanipekun, 2007). In this study, lead was

accumulated in the decreasing order of gill > liver > muscle. High concentration of metals in the gill has often been used as

an indication of acute exposure since the metals are fixed by absorption processes which occur very rapidly (Oladimeji and

Offem, 1989; Noeqrohati, 2006). It has also been reported that much of lead can be bound externally (Hares et al., 1991).

The larger surface area of the gills in contact with the medium then could probably account for the higher concentration of

lead in the gill compared with the liver and the muscle in this study. This is more likely since the primary source of lead to

the body is from the water via the gills. Closely associated with the gills are iron oxides involved in the carriage of

respiratory gases. The presence of iron oxide in the gills is known to enhance lead disposition (Hare et al., 1991). Lead isknown to strongly absorb onto iron oxides but not highly assimilated (Hare et al., 1991). In this study, a distinction was not

made between lead absorbed on the gill externally and those bound internally.

CONCLUSIONS

The general increase observed in the level of lead with increased exposure period might be due to increase in the

level of low molecular weight metal-binding proteins such as hepatic metallothionein. Such an increase might be a

response by the fish to remove lead from the circulation and hence reduce lethal effects. Low molecular weight metal

binding proteins are known to play a significant role in the detoxication of heavy metals (Daramola and Oladimeji, 1989).

The increase in lead level might be due to a negative feedback mechanism whereby more lead enters the tissue andbecomes bound to the metallothinnein. If this should happen, then the lead burden in the fish will increase with increased


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exposure time. Increase was also recorded in the level of lead in the control fish. This increase was however significantly

lower than the lead burden reported in the two highest concentrations of exposed fish. Since lead was not introduced into

the control, lead in the control fish can only come from the feed with which they were fed or the well water to which they

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Documents

6. Ijeefus - Lead Accumulation Profiles in the Soft Tissues of The