7. Ijeefus -Accumulation of Lead

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ACCUMULATION OF LEAD IN THE TISSUES OF FRESHWATER

PSEUDAPOCRYPTES ELONGATUS EXPOSED TO STATIC NOMINAL

CONCENTRATIONS OF LEAD NITRATE

SUBHAMOY DAS1, SANJOY MAJI 2, DEBNARAYAN ROY 3,

RAJKUMAR GUCHHAIT 4 & AVISHEK DOLAI 5

1,2,4Department of Zoology, Mahishadal Raj College, Mahishadal, West Bengal, India 3Department, of Zoology, A. B. N. Seel College, West Bengal, India 5Department of Zoology, University of Calcutta, West Bengal, India

ABSTRACT

Static bioassay methods were used under laboratory conditions to determine the accumulation of lead by

Pseudapocryptes elongatus. After various time (10,20, 30 And 40 days) of exposure, to100. 200, 300, 400 and 500 ppm of

lead nitrate different tissue of Pseudapocryptes elongatus (brain, kidney, liver, skin, gill and muscle) accumulated lead

was remarkably different. Finally it will conclude that in fish P. elongatus the accumulation of lead the gill stood first,

followed by liver, followed by brain, kidney and skin. If we remove the gill of target fish our body will be safe at any

concentration of lead and at any time of exposure period.

KEYWORDS: Bioaccumulation, Pseudapocryptes Elongatus Heavy Metal, Lead, Sublethal, Liver, Skin, Brain, Kidney,

Muscle Kidney

Received: Nov 07, 2015; Accepted: Nov 20, 2015; Published: Dec 11, 2015; Paper Id.: IJEEFUSDEC20157

INTRODUCTION

Among various toxic metal which were released from various industry, lead is one of the major element.

These were the chemicals which were released from different battery industry, fertilizer industry and refining ores

(Handy, 1994). According to various scientist, they mentioned that gasoline and fishery boats also the other major

sources of the lead pollution. Lead in the environment present in the both organic and inorganic form but inorganic

form is the major form which is the soluble in acidic water that is in the lower pH range (Moore and Rainbow,

1987). Lead enters in the tissue of fish as well as other organism in various rout. The above lead may enter through

two major rout that is directly from the surrounding and through diet of the targeted organism (Bury et al., 2003).

Along with this rout fish accumulate through the food which absorbed in the intestine. (Kotze et al., 1999; Ay et

al., 1999; Macdonald et al., 2002, Hensen et al., 2007). There are various report regarding the relation between,

rate of accumulation and ecological factors that is amount of accumulation and temperature salinity, time of

exposure pH, storage in the tissue, rate of elimination and rate of uptake. (Roesijadi and Robinson, 1994;

Langston, 1990). Various worker reports the amount of accumulation in different tissue in various organ, that is

liver, brain, kidney, muscle and kidney (WHO, 1980; Chen and Chen, 2001; Allinson et al., 2002; Spokas, et al.

2006; Fabris et al. 2006).Lead is a toxic metal which disrupt the function of various parts of brain(Rademacher et

al. 2003) which result into loss of nervous coordination and also cause for cancer(Eisler 1988).In the present

investigation, a study was conducted to estimate the uptake and accumulation of waterborne Pb in various tissues

Original A

rticle International Journal of Environment, Ecology, Family and Urban Studies (IJEEFUS) ISSN(P): 2250-0065; ISSN(E): 2321-0109 Vol. 5, Issue 6, Dec 2015, 51-62 © TJPRC Pvt. Ltd.

52 Subhamoy Das, Sanjoy Maji, Debnarayan Roy, Rajkumar Guchhait & Avishek Dolai

Impact Factor (JCC): 3.0965 NAAS Rating: 3.63

like skin, gills, eyes, liver, intestine and muscles of fingerlings of a freshwater fish, Pseudapocryptes elongates.

Pseudapocryptes elongatus is a demersal and amphidromous species which is found in mudflats of estuaries and the

freshwater tidal zone of rivers. P. elongatus is a amphibious air-breather.

Fish Pseudapocryptes elongatus is distributed from the east coasts of India to South East Asia; its presence is not

confirmed in China and one single record is available from Kyushu Is., Japan.Asia - Inland waters; Bangladesh; Cambodia;

Chilka Lake; China; Gulf of Thailand; India; Indian Ocean; Eastern; Indonesia; Indonesian Sea; Indo-Pacific: India to

Tahiti and north to China.

Fish Pseudapocryptes elongates is very popular benthic freshwater edible fish which is benthic in habit and

carnivorous in nature. Major food of this fish is small fish, small invertebrate, small prawn. In the laboratory it can survive

by consuming tubifex and artificial dry food. It can collect food by various method such as suspended collection method,

filtering method and mud scraping method. 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.

Pseudapocryptes elongatus or gulee or Chemma fish is commercially important fresh water benthic fast moving

mud duealing fish species in wet land of west Bengal and surrounding.. The Pseudapocryptes elongatus is an carnivore and

usually inshore, entering pond, canal, wetland, jheel, river and lagoons strictly having slightly saline in nature. This fish is

no doubt is estuarine fish. Pseudapocryptes elongates is surface breather. Pseudapocryptes elongatus is considered to be

Least Concern since it is very widely distributed and with limited threat. Pseudapocryptes elongatus is widely distributed

throughout India, Pakistan, Nepal, and Bangladesh. Present in most of the Ganges drainage,. In West Bengal its main

population were found at Kakdwip, Canning, Dimond Harbour, Nayachar, Geonkhali, Basanti, Gosaba and other estuarine

site having mud and semi saline water. Pseudapocryptes elongatus occurs in running and stagnant waters. Found in

brackish waters and deltas of large rivers, common in ponds and slow flowing rivers with brackish water. Inhabits still

waters with silt or mud substrate. Nocturnal feeder, on insects and worms in lowland habitats and at moderate elevation in

all the larger river systems of the Indian subcontinent.

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, Pseudapocryptes

elongatus, specially in the ecological condition of West Bengal.As heavy metal can change various metabolic pathway at

high concentration they can cause serious health hazard(Tort et al 1987). It also cause biological amplication (Unlu 1993).

The mechanism of action of heavy metal were well explained by various scientist (Holcombe et al 1976, Barlas 1999,

Guerrin et al 1990). Pseudapocryptes elongatus is considered a good test organism for heavy metal contamination because

of its feeding behavior and bottom feeding habits. 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 carried out to observe the

rate of accumulation of heavy metal Pb in different tissue or organ in the body of Pseudapocryptes elongatus.

In the present investigation, a study was conducted to estimate the uptake and accumulation of waterborne Pb in

various tissues like skin, gills, eyes, liver, intestine and muscles of fingerlings of a freshwater fish, Pseudapocryptes

elongatus.

Accumulation of Lead in the Tissues of Freshwater Pseudapocryptes 53 elongatus Exposed to Static Nominal Concentrations of Lead Nitrate


MATERIALS METHODS

Fish Collection and Acclimatization

Fish Pseudapocryptes elongatus were collected from different sites at Uluberia, Kakdwip, Basanti, Nayachar,

Terapakhya, Canning, Dimond Harbour, Haldia and Nayachar Island of West Bengal (Figure 1) 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.

Morphometric Analysis

The Pseudapocryptes elongatus adult used in this study weighs between 18.60g and 24.40g (mean weight =

21.45g). They were brought to the laboratory in polythene bags. In the laboratory, they were acclimated in aquaria using

tap water maintained at 26.0C for few week. They were kept in special technique. The water mixed with some soil. The

depth of soil will not exceed 4cm depth.

Food and Maintenance

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 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 the heavy metals lead, the

experiments were designed accordingly.

Analysis of Physico Chemical Parameters

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 physicochemical

parameters of the water were determined before use in the experiments using standard methods.

Determination of LC 50

For determination of the 96-h LC50 (lethal concentration) values, various concentrations each of lead nitrate (E

MARK) were used. A group of 8 fish was used for each concentration of each heavy Pseudapocryptes elongatus 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 650.0 ppm for lead

nitrate. The LC50 values were found and the true concentration of lead was obtained

Stock Solution Prepatation

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 nitrate 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 Pseudapocryptes elongatus were randomly loaded into

100 liter glass aquaria containing the different concentration of lead nitrate. 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 40

days.

Fish samples were taken after 10, 20, 30 and 40 days (after this duration all the fish irrespective to dose not

survive at all.) from the different concentrations and the control group. The organs were isolated and weighed separately.

Main organ 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-ashed 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 for lead using Atomic Absorption Spectrophotometer. The lead residues in each

sample were calculated based on FAO/SIDA (1983) methods.

Sampling

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 1000 mm which occurs in the months of February – April 2015. The Pseudapocryptes elongatus

samples were caught by trap anchored using locally made hoop-type 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. 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 de-ionized

water at the Centre. A clean high quality rush free and corrosion resistant stainless steel knife was used to cut about 2 g of

muscle 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, skill, brain and gills of all the Pseudapocryptes elongatus 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 clean



polyethylene 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-ionized water.

These solutions were then analyzed using a Atomic Absorption Spectrophotometer model AES/ 23908/RB/DER-.

Calibrations, precision together with accuracy of this system has previously been done by the authority.

RESULTS

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

bioaccumulation experiment was presented in Table 12. 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 13 DO2, alkalinity, salinity,

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

Table 1 shows the Amount of bioaccumulation in different organ of Pseudapocryptes elongatus against exposure

in 100ppm lead nitrate after different period (days) of exposure. Table 2, 3, 4,, and 5 shows the Amount of bioaccumulation

in different organ of Pseudapocryptes elongatus against exposure in 200, 300 400 and 500 ppm ppm lead nitrate after

different period (days) of exposure. Table 6: Table : Amount of bioaccumulation of lead nitrate in brain of Pseudapocryptes

elongatus against exposure in different doses after different period (days) of exposure. Table 7: Table : Amount of

bioaccumulation of lead nitrate in skin of Pseudapocryptes elongatus against exposure in different doses after different

period (days) of exposure. Table 8: Table: Amount of bioaccumulation of lead nitrate in liver of Pseudapocryptes elongatus

against exposure in different doses after different period (days) of exposure. Table 9: Table : Amount of bioaccumulation of

lead nitrate in muscle of Pseudapocryptes elongatus against exposure in different doses after different period (days) of

exposure. Table 10: Table : Amount of bioaccumulation of lead nitrate in kidney of Pseudapocryptes elongatus against

exposure in different doses after different period (days) of exposure. Table 11 Table : Amount of bioaccumulation of lead

nitrate in gill of Pseudapocryptes elongatus against exposure in different doses after different period (days) of exposure.

Bioaccumulation of lead in different tissue of Pseudapocryptes elongatus after different exposure period at 100 ppm lead

nitrate concentration indifferent organ.

Table 1: Amount of Bioaccumulation in Different Organ of Pseudapocryptes elongatus against Exposure in 100ppm Lead Nitrate after Different Period (Days) of Exposure

Organs Exposure Time(day)

10 20 30 40 Brain 0.1 0.1 0.027 0.06 Liver 0.015 0.063 0.32 0.9 Gill 0.31 1.07 1.09 1.65



Table 1: Contd., Skin 0.05 0.09 0.407 0.49 Muscle 0.03 0.23 0.467 0.32 Kidney 0.23 0.295 0.275 0.32

Table 2: Amount of bioaccumulation in Different Organ of Pseudapocryptes elongatus against

Exposure in 200ppm Lead Nitrate after Different Period (Days) of Exposure

Organs Exposure Time(day)

10 20 30 40 Brain 0.2 0.31 0.087 0.0900 Liver 0.02 0.097 0.689 1.5760 Gill 0.42 3.34 1.895 2.0000 Skin 0.055 0.318 0.515 0.6340 Muscle 0.69 0.58 0.59 0.6680 Kidney 0.27 0.385 0.39 0.3300

Table 3: Amount of Bioaccumulation in Different Organ of Pseudapocryptes elongatus against

Exposure in 300ppm Lead Nitrate after Different Period (Days) of Exposure

Organs Exposure Time(Day)

10 20 Brain 0.2230 0.365 Liver 0.0300 5.5600 Gill 0.4200 5.8900 Skin 0.1900 0.3850 Muscle 0.7100 0.7540 Kidney 0.3150 0.4030

Table 4: Amount of Bioaccumulation in Different Organ of Pseudapocryptes elongatus against

Exposure in 400ppm Lead Nitrate after Different Period (days) of Exposure

Organs Exposure Time(Day)

10 20 Brain 9.6000 11.23 Liver 0.0360 8.4 Gill 0.4050 6.1 Skin 0.2050 0.94 Muscle 0.1100 0.75 Kidney 0.3778 0.45

Table 5: Amount of Bioaccumulation in Different Organ of Pseudapocryptes elongatus

against Exposure in 500ppm Lead Nitrate after 10 Days Exposure

Organ Amount of Lead(ppm) Brain 11.2000 Liver 0.1420 Gill 0.6900 Skin 0.2650 Muscle 0.2110 Kidney 0.3900

Table 6: Amount of Bioaccumulation of Lead Nitrate in Brain of Pseudapocryptes elongatus against Exposure in Different Doses after Different Period (Days) of Exposure

Exposure Time(Day)

Doses of Lead Nitrate(ppm) 100 200 300 400 500

10 0.1 0.2 0.2230 9.6000 11.2000



Table 6: Contd., 20 0.1 0.31 0.365 11.23 NA 30 0.027 0.087 NA NA NA 40 0.06 0.0900 NA NA NA

Table 7: Amount of Bioaccumulation of Lead Nitrate in Skin of Pseudapocryptes elongatus

against Exposure in Different Doses after Different Period (Days) of Exposure

Exposure Time(Day)


10 0.05 0.055 0.1900 0.2050 0.2650 20 0.09 0.318 0.3850 0.94 NA 30 0.407 0.515 NA NA NA 40 0.49 0.6340 NA NA NA

Table 8: Amount of Bioaccumulation of Lead Nitrate in Liver of Pseudapocryptes elongatus


Exposure Time(Day)


10 0.015 0.02 0.0300 0.0360 0.1420 20 0.063 0.097 5.5600 8.4 NA 30 0.32 0.689 NA NA NA 40 0.9 1.5760 NA NA NA

Table 9: Amount of Bioaccumulation of Lead Nitrate in Muscle of Pseudapocryptes elongatus


Exposure Time(Day)


10 0.03 0.69 0.7100 0.1100 0.2110 20 0.23 0.58 0.7540 0.75 NA 30 0.467 0.59 NA NA NA 40 0.32 0.6680 NA NA NA

Table 10: Amount of Bioaccumulation of Lead Nitrate in Kidney of Pseudapocryptes elongatus


Exposure Time(Day)


10 0.23 0.27 0.3150 0.3778 0.3900 20 0.295 0.385 0.4030 0.45 NA 30 0.275 0.39 NA NA NA 40 0.32 0.3300 NA NA NA

Table 11: Amount of Bioaccumulation of Lead Nitrate in Gill of Pseudapocryptes elongatus

against Exposure in Different Doses after Different Period (days) of Exposure

Exposure Time(Day)


10 0.31 0.42 0.4200 0.4050 0.6900 20 1.07 3.34 5.8900 6.1 NA 30 1.09 1.895 NA NA NA 40 1.65 2.0000 NA NA NA



Table 12: Morph Metric Measurement of the Fish Pseudapocryptes elongatus, (at Different Sites) Used for the Bioaccumulation

Site Mean Length in

cm(Aver, Range, SE±) Mean Weight in G (aver, Range, SE±)

Month Moisture Content in

%( Aver, Range, SE± ) Nayachar 16.25 (9.1-19.01,±3.22 14.7, 7.23-17.8,±2.98 19-Mar 23,22±4.43 Geonkhali 14.99(10.22-17.55, ±1.89) 11.9, 8.23-11.69, 31.35 24-Mar 86.98±2.61 Kakdwip 12.21 (8.45-13.34,) ±1.33 11.23, 7.29-9.87, ±1.33 17-Apr 84.34±2.98

Table 13: Physico chemical parameters of the water, habitat at kakdwip of the

Pseudapocryptes elongatus, used for bioassay and bioaccumulation experiments.

Experiment. Parameters Range Mean Colour TCU 79.00 – 171.60 134 ±12.22 Temperature 0C 25.6 – 27.50 25.40 ± 1.221 pH 6.1 – 7.4 6.6 ±064 Salinity ‰ 7.36 –9.40 8.50 ± 1.51 Dissolved Oxygen (ppm) 6.96 – 7.21 6.7 ± 0.42 Settleable solids 0.39 – 1.52 0.62 ± 0.15 Alkalinity (ppm) 24.36 – 38.54 33.64 ± 2.05 Total hardness as CaCO3 (ppm) 72.25 –110.60 92.56 ± 3.90 Biological Oxygen Demand (BOD) (ppm)

18.65 – 29.22 23.98 + 3.48

Chloride (ppm) 6.60 – 9.2 8.42 ±1.321 Sulphate (ppm) 0.00 – 1.20 0.06 ± 0.044 Nitrate (ppm) 2.85 – 4.96 4.05 ± 0.47 Phosphate (ppm) 0.02 – 0.68 0.24 ± 0.19 Ammonia (ppm) 0.01 – 0.05 0.01 ± 0.025 Copper (ppm) 0.01 – 0.03 0.01 ± 0.081 Lead (ppm) 0.190 –0.71 0.656 ± 0.24 Iron (ppm) 0.50 – 1.64 0.76 ± 0.64 Zinc (ppm) 0.01 – 0.03 0.013 ± 0.011 Chromium (ppm) 0.03 – 0.06 0.031 ± 0.017 Manganese (ppm) 0.01 – 0.19 0.03 ±0.017

DISCUSSIONS

The accumulation of heavy metals by aquatic organisms involves tissues that serve as the site for uptake and

absorption like gills, skin and intestine. Significant Pb accumulation occurred in the skin (including mucus) and was

followed by liver, gills, eyes, muscles, and intestine. Skin, gills and eyes are tissues which were directly exposed to

waterborne Pb have showed high uptake and accumulation which increased significantly high with the passage of time.

The Pb uptake and accumulation in these tissues was highest at higher concentration with longer exposure whereas

minimum Pb uptake and accumulation was observed at lower dose during the first week of exposure. This is because high

dose and longer exposure resulted into high uptake and accumulation.

The result about Amount of bioaccumulation in different organ of Pseudapocryptes elongatus against exposure in

100ppm lead nitrate after different period (days) of exposure shows that in 10 days exposure, gill accumulate

maximum(.31) followed by kidney(.23), brain(.1), skin(.05) and liver(.015). When exposure in 40 days period was

estimated, also will shows maximum(1.65). But in comparison with 10 days period accumulates 5 fold higher lead in the

gill tissue. But very interesting to note that the amount of lead in brain reduce after 10 days exposure. I 10, 20, 30 40 days

exposure the lead is.1,.1,.02,.06. What is the actual mechanism? In the brain tissue the amount of fat is higher. It may affect

reason to release the lead particle somehow. In liver within 40 days the deposition increased 6 fold ie, from,.015 to.9ppm.



Although skin accumulates less still its rate of accumulation increase 10 folds. The same result were also found in muscle.

Regarding kidney the accumulation increased very little amount ie,.23 to.32.

In Table 2 the Amount of bioaccumulation in different organ of Pseudapocryptes elongatus against exposure in

200ppm lead nitrate after different period (days) of exposure was presented. It shows that brain, liver, gill, skin, muscle and

kidney the accumulation of lead occur after 10 days was.2,.02,.42,.05,.69,.27 respectively. It shows that brain, liver, gill,

skin, muscle and kidney the accumulation of lead occur after 20 days was.31,.09, 3.3,.34,.58,.38 respectively. In case of

brain, liver, gill, skin, muscle and kidney the accumulation of lead occur after 30 days was.087, 1.5, 2.0,.6,.6 and.33

respectively. After 40 days the accumulation increased in brain, liver, gill, skin, muscle and kidney in 4, 25, 5, 12, 9 and 1.5

folds. So after 40 days at 200 ppm exposure liver shows maximum fold increase and kidney shows the least fold of

increase.



kidney the accumulation of lead occur after 10 days was.22,.03,.4,.19,.71,.31 respectively. It shows that brain, liver, gill,

skin, muscle and kidney the accumulation of lead occur after 20 days was.36, 5.5, 5.8,.38,.75 and.41 respectively. In case

of brain, liver, gill, skin, muscle and kidney the accumulation of lead occur after 30 days was not measured as the fish can

not survive it that doses.. After 20 days the accumulation increased in brain, liver, gill, skin, muscle and kidney in 1.5, 132,

1.7, 4, 1, 1.23 folds. So after 20 days at 300 ppm lead nitrate exposure liver shows maximum fold increase and kidney

shows the least fold of increase.



kidney the accumulation of lead occur after 10 days was 9.6, 8.0,.40,.20,.11,.3ppm respectively. It shows that brain, liver,

gill, skin, muscle and kidney the accumulation of lead occur after 20 days was 11.23, 8.4, 6.1,.94,.75,.45ppm respectively.

In case of brain, liver, gill, skin, muscle and kidney the accumulation of lead occur after 30 days was not measured as the

fish cannot survive it that doses. After 20 days the accumulation increased in brain, liver, gill, skin, muscle and kidney in

1.6, 20. 4.5, 7 and 1.5 folds. So after 20 days at 400 ppm lead nitrate exposure liver shows maximum fold increase and

kidney shows the least fold of increase.

In Table 5 the Amount of bioaccumulation in different organ of Pseudapocryptes elongatus against exposure

in5300ppm lead nitrate after different period (days) of exposure was presented. It shows that brain, liver, gill, skin, muscle

and kidney the accumulation of lead occur after 10 days was 11.2,.14,.69,.49,.29, and.29 respectively. It shows that brain,

liver, gill, skin, muscle and kidney the accumulation of lead occur after 20, 30 and 40 days were not measured as the fish

can not survive it that doses. So after 10 days at 500 ppm lead nitrate exposure liver shows maximum fold increase and

kidney shows the least fold of increase.

In Table 6 the Amount of bioaccumulation of lead nitrate in brain of Pseudapocryptes elongatus against exposure

in different doses after different period (days) of exposure was presented. It shows at 10, 20, 30 and 40 days exposure

against brain, at 100 ppm, the lead was.1,.1,.02,.06ppm respectively. It shows at 10, 20, days exposure against brain, at 200

ppm, the lead was.2,.31,.08,.09ppm respectively. It shows at 10, 20, days exposure against brain, at 400 ppm, the lead was

9.6, 11.23ppm respectively. It shows at 10, days exposure against brain, at 500 ppm, the lead was 11.20 ppm.



In Table 7 the Amount of bioaccumulation of lead nitrate in skin of Pseudapocryptes elongatus against exposure


against brain, at 100 ppm, the lead was.05,.09,.40,.49 ppm respectively. It shows at 10, 20, days exposure against brain, at

200 ppm, the lead was.05,.31,.51,.63ppm respectively. It shows at 10, 20, days exposure against brain, at 400 ppm, the lead

was.13,.38 ppm respectively. It shows at 10, days exposure against brain, at 500 ppm, the lead was.26 ppm.

In Table 8 the Amount of bioaccumulation of lead nitrate in liver of Pseudapocryptes elongatus against exposure


against skin, at 100 ppm, the lead was.01,.06,.32,.9 ppm respectively. It shows at 10, 20, days exposure against skin, at 200

ppm, the lead was.03, 5.5ppm respectively. It shows at 10, 20, days exposure against skin, at 400 ppm, the lead was.03, 8.4

ppm respectively. It shows at 10, days exposure against skin, at 500 ppm, the lead was.14 ppm.

In Table 9 the Amount of bioaccumulation of lead nitrate in muscle of Pseudapocryptes elongatus against

exposure in different doses after different period (days) of exposure was presented. It shows at 10, 20, 30 and 40 days

exposure against muscle, at 100 ppm, the lead was.03, 2.3,.46,.32 ppm respectively. It shows at 10, 20, days exposure

against muscle, at 200 ppm, the lead was.69,.58,.59,.66ppm respectively. It shows at 10, 20, days exposure against muscle,

at 400 ppm, the lead was.11,.75 ppm respectively. It shows at 10, days exposure against muscle, at 500 ppm, the lead

was.21 ppm.

In Table 10 the Amount of bioaccumulation of lead nitrate in kidney of Pseudapocryptes elongatus against

exposure in different doses after different period (days) of exposure was presented. It shows at 10, 20, 30 and 40 days

exposure against kidney, at 100 ppm, the lead was.23,.29,.27,.32 ppm respectively. It shows at 10, 20, days exposure

against kidney, at 200 ppm, the lead was.27,.38,.39,.33 ppm respectively. It shows at 10, 20, days exposure against kidney,

at 400 ppm, the lead was.31,.4 ppm respectively. It shows at 10, days exposure against kidney, at 500 ppm, the lead was.39

ppm.

In Table 11 the Amount of bioaccumulation of lead nitrate in gill of Pseudapocryptes elongatus against exposure


against gill, at 100 ppm, the lead was.31, 1.0, 1.09, 1.65 ppm respectively. It shows at 10, 20, days exposure against gill, at

200 ppm, the lead was.42, 3.3, 1.8, 2.0ppm respectively. It shows at 10, 20, days exposure against gill, at 400 ppm, the lead

was.40. 6.1 ppm respectively. It shows at 10, days exposure against gill, at 500 ppm, the lead was.69 ppm.

The amount od accumulated lead may be caused by various physic chemical as well as biological parameters

(Kotze et al. 1999) My study have revealed that Pb level in the water of river is 2.4-3.55 ppm (more than 50 times higher

than WHO standards for drinking water) and the fish species including P. elongates had accumulated very high level of Pb

(31.2 – 38.3 ppm) in the muscles which is more than 600 times

CONCLUSIONS

Finally it will conclude that in fish P. elongaus the accumulation of lead the gill stood first, followed by liver,

followed by brain, kidney and skin. The sequence is gill> kidney > brain > kidney > skin. If we remove the gill of target

fish our bode will be safe at any concentration of lead and at any time of exposure period. More over it is interesting to

note that in brain the accumulation is cooperatively less than other report. The amount of deposition in skin is least. It is

safe to consume the skin of the fish P. elongates. Moreover at last it should be stated that this study need further detail



report as the accumulation depends on age, sex, ecological condition.

ACKNOWLEDGEMENTS

We thankfully acknowledge to Principal, Mahishadal Raj College, Purba Medinipur, for providing us the facility

of the above work. We also thankful to U G C to provide the fund for the above analytic work.

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7. Ijeefus -Accumulation of Lead