| Research Article (Open Access) |
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ABSTRACT-
Ramgarh taal is an important water body of Gorakhpur, U.P, India, which received untreated effluents
from different sources of Gorakhpur city. The aim of the present study was to evaluated the toxic impact of these effluents
on water quality of water and its toxic effect on fish population present in this lake with two sets of experiments (1) Fish
caged in pollution free water i.e. control group and (2) Fish directly caught from different sites of water body. The
effluents discharged in this water body caused significant alterations in pollution parameters of water and significantly
altered the biochemical profile (total amino acid levels increased while total protein, Glycogen and nucleic acid levels
decreased from the levels of control group) and hematological parameters (PLTs, WBCs, RBCs, HGB, HCT, MCV,
MCHC etc) decreased from the levels of control group. Seasonally variations in Physico-chemical parameters
(Temperature, pH, DO, BOD and COD) of water body also measured. Which were much higher than the tolerance limit
recommended by General standard by Central Pollution Control Board of India.
Key-Words- Ramgarh taal, Parameters, Biochemical parameters, Amino acid, Hematological, Physiological, Pollution,
Pesticides, Herbicides, Channa punctatus
INTRODUCTION-
Environmental pollution is a worldwide problem because
human activities are get polluted environment. Day by day
Environmental contaminates are attributed to producing of
different products, are release many toxic chemicals to
capable of interacting aquatic or terrestrial ecosystem.
Many types of toxic substance are lipophilic and weren’t
adversely affected by water [1] i.e. they are easily able to
performed for penetrate cell membrane and frequently
posses high bioaccumulation factors. The effluent discharge
from industrial waste, agricultural waste, sewage, that is
directly drainage into water bodies without treatment they
constitute biohazards to man and other living organism
such as aquatic animals because they contain toxic
substances detrimental to serious effects on health [2-3]. Since much effluent discharged on ground or in water
bodies and they are not treated properly results these
effluents rapidly accumulated in the system of water bodies
through food chain [4]. Fish are important members of
aquatic ecosystem but now days they get more polluted
because different effluent sources are directly drainage in to
reservoir water and affect on aquatic flora and fauna as well
as human health [5]. Many water bodies such as Lake,
pond, river etc have different types of flora and fauna they
cannot avoid exposure to these toxic effluents and chemical
which used in the form of fertilizer for well production of
crops, that suspended or dissolved these water bodies thus
that living organism move to favorable condition to avoid
unfavorable condition [6]. Industrial waste is the most
major types of effluent which polluted environmental
system. The fresh water bodies are being affected by
exclusive influence of human activities [7]. Thus it is truly
stated that the Ramgarh Lake is more polluted because it
receives huge amount of effluent sources from agricultural
waste, house hold material, automobiles washing waste etc
from different sites. The aim of this study was evaluated the
effects of pollutants on flora and fauna of Ramgarh Lake
and how much effects economically on fisheries as well as
this data provide awareness baseline information for people settled around the lake and government to take action for
treatment polluted water of Ramgarh Lake.
MATERIALS AND METHODS-
Discription about study area and effluents
discharged-
The present study area is Gorakhpur situated in the east of
Uttar Pradesh, India on National highway-28 lies between
Lattitude 26ş 46’ N and Longitude 83ş 22’ E, cover the
geographical area of 3483.8 sq. Ramgarh Lake is a large
natural lake situated southeast of Gorakhpur in
eastern Uttar Pradesh. It lies within the floodplain of Rapti
River and outflow into it through a drain called Gurrah
Nalla. The lake had maximum water spread of about 723
hactares in 1970s but has now shrunk to 678 hactares. It
maximum water depth has also decline from 4.5 m in 1990s
to less than 3.5m at present. Now day Ramgarh Lake has
more polluted because it receives huge amount of effluent
discharge from different sites and its effects on flora and
fauna of lake. These pretreated effluents from the
remainder of the different sources are collected into a single
drain and release into the Ramgarh Lake.
PHYSIOCHEMICAL STATUS
Collection and analysis of polluted water sample-
Effluents and polluted water samples were collected from
different sites of Ramgarh lake in glass stopper bottle at
undisturbed stage from the three part of the each selected
sites –
A- Entry point of effluent in the lake
B- 200 meters away from the entry point in downstream
C-200 meters away from the entry point in upstream in
summer, rainy and winter season
Sampling was done from the ranging from 25 to 50 cm at
various points. Care was taken to avoid any disturbance by
loose sediments. The collected samples transported
immediately to the laboratory and the physical and
chemical characteristic of polluted water samples estimated
(Table-1). The samples were collected analyzed for
temperature, pH, dissolve oxygen (DO), biological oxygen
demand (BOD) and chemical oxygen demand (COD)
values on the basis of APHA [8] method.
BIOCHEMICAL STUDY
Collection of fish-
10-15 healthy Channa punctatus (commonly snake head)
of an average total length of 170.57 cm and average weight
of 1002.88 gm were brought to laboratory from Gorakhpur
hatchery Chhapia, Gorakhpur (U.P.), India. They were
maintained for acclimatization for two–three weeks in
aerated aquarium of 45 liter capacity. These acclimatized
fish are used for control group as well as brought polluted
fish from selected site of Ramgarh lake and it directly used
for biochemical analysis i.e. uncontrolled group.
Commonly snake head (Channa punctatus) is member of
the Channidae family. It is identified by body elongated and
cylindrical. Eyes are comparatively small and located
anterior part of head and lower jaw slightly protruding,
absent barbells.
Experimental design-
Potential effect of effluents was studied in the form of
biochemical parameters (protein, total free amino acid,
glycogen and nucleic acid) in the set of two experiments:
· Fish caged in pollution free water i.e. control
· Fish directly caught from Ramgarh Lake.
Analysis of Biochemical parameters-
After completion of experiment the caged fish and captured
fish were brought to the laboratory and washed with water
and killed by severe blow on head and operated muscles
and liver tissue quickly dissected out in ice tray and used
for biochemical analysis.
Protein-
Protein levels were estimated according to the method of
Lowery [9] using standard solution of bovine serum
albumin.10% TCA using for homogenates liver and muscle
tissues.
Total Free Amino Acids-
Estimation of total free amino acid on the basis of Spice
[10] method. Homogenates 50mg tissues were prepared
in95% ethanol, centrifuged at 6000xg and used for amino
acid estimation.
Glycogen-
Glycogen was estimated by the Anthrone method of Van
Der Vies [11]. In the present experiment 50 mg of tissue
were homogenates with 5ml of TCA. The homogenate was
filtered and 1.0 ml filtered was used for assay.
Nucleic Acid-
Observation of nucleic acid (DNA and RNA) was
performed, by methods of Schneider [12]. Using
diphenylamine and Orcinol reagents respectively.
Homogenates tissue was prepared in 5% TCA at 900 C,
centrifuged at 5000xg for 20 min and supernatant was
prepared and used for estimation. Both DNA and RNA
have been expressed as µg/mg tissue.
Hematological experiment-
For the study of hematological, taken 2 healthy fish (each
site) about 20 cm length and 200g weight. Blood samples
of approximately 4 ml were taken from each by puncturing
caudal vein with sterile heparinized syringes and needles.
Samples were taken into labeled Eppendorf tubes
containing 0.5 mg ethylene diamine tetra acetic acid
(EDTA) an anticoagulant; it was properly mixed and used
for hematological analysis for all hematological parameters such as red blood cell (RBC), count, the total white blood
cell count (WBCs), hemoglobin percentage (Hb%),
Erythrocytes sedimentation rate (ESR) and the packed cell
volume (PCV), the mean corpuscular volume (MCV), the
mean corpuscular hemoglobin (MCH) and the mean
corpuscular hemoglobin concentration (MCHC), in the
natural product laboratory of D.D.U Gorakhpur University
for hematological experiment by an Autoanaliser (Celltac
a) NIHON KOHDEN.
RESULTS-
The data represented are discussed on the basis of three
seasons i.e. summer, rainy and winter. The results sof
physico-chemical parameters of water samples and analysis
of hematological and biochemical parameters of fish caged
in unpolluted water body and polluted fish directly caught
from different sites of Ramgarh Lake are given in Table-1,
Table-2, and Table-3 respectively.
Physico-chemical parameters study-
The results of physico-chemical analysis of effluent water
samples from selected sites are given in Table-1. The data
described on the basis of three seasons i.e. summer, rainy
and winter. This study has been done during May
2015- December 2015.
Temperature of effluents water samples varied with
different seasons. The highest temperature observed in
summer season at site-1 which is 30.0±0.03 and lowest
temperature 15.6±0.07 in winter season at site-2. The pH
value measure of the acidity and alkalinity of water and is
one of most stable measurements. Highest value of Ph
observed in all three season at site-1 i.e. 7.6±0.10-8.5±0.06.
Dissolved oxygen (DO) is important for water bodies
animal and during study of research work lowest DO in all
sites with the comparison of central pollution control board
of India. Measurement Biological oxygen demand (BOD)
during all three season find that the highest value of BOD
in site-1 and lowest in site-3 as well as the Chemical
oxygen demand (COD) maximum in summer, rainy and
winter at site-1 which is 1126.6±1.46, 1180±2.886 and
1210±2.856 respectively and minimum at site-3whch is
425±2.145, 291±4.509 and 254±1.527of summer, rainy and
winter respectively. But both BOD and COD has been
highest value in all sites in all season as comparison to
central pollution control board of India.
Hematological Observation-
The effects of effluents on haematological parameters of
Channa punctatus was studied. Haematological parameters
of polluted fish and those fish were caught from unpolluted
area i.e control group showed significant (p<0.05)
difference. The effluents from different sites are highly
toxic for flora and fauna. During hematological, it was
observed that the highly decrease level of WBCs and RBCs
from the level of control group (Table-2). Results
concerning red blood cells counts exhibited a decrease in their number, mean corpuscular haemoglobin concentration
(MCHC), mean corpuscular haemoglobin (MHC), mean
corpuscular volume (MCV) were significantly decrease
than control groups. The level of PLT, WBCs, and RBCs of
site-1 is lowest as comparison to site-2 and site-3. Thus
results is obtained that site-1 is more polluted area as
comparison to site-2 and site-3.
Bio-chemical Observation-
The effluents discharged from site-1, 2, and 3 in Ramgarh
Lake, its caused serious effects on flora and fauna. During
biochemical experiment results observed that the
significant alteration in protein, free amino acid, glycogen
and nucleic acid in liver and muscles tissue of fish Channa punctatus. Total protein was reduced to 27%, 30%, 34%,
35%, 42%, 34% of control, nucleic acid level such as DNA
was reduced to 48%, 26%, 43%, 33%, 52%, 48% of
control, similarly, RNA level was reduced to 40%, 45%,
48%, 35%, 40%, and 43% of control. Glycogen level was
reduced to 64%, 48%, 67%, 44%, 755and 78% of control,
while total free amino acid was induced to 530%, 545%,
496%, 496%, 325% and 538% of control respectively in
liver and muscle tissue in summer, rainy and winter season
of site-1 (Table- 3). Similarly trend of result were observed
for site-2 (Table- 3). The value of all biochemical
parameters of site-3 is up to above 60% - 80% of control,
its mean approximately same to control group, so it is clear
that the fish collected from the site-3 is less polluted to
effluents than site- 1 and site- 2.
Table 1: Physico-chemical characteristic of polluted water samples collected from different sites of Ramgarh lake
in summer (May to June 2015), rainy (August to September 2015) and winter (November to December 2015)
| Characteristic | Season | Site-1 | Site-2 | Site-3 | General standard by central pollution control board of India |
|---|---|---|---|---|---|
| Temperature | Summer | 30.0±0.03 | 29.9±0.09 | 29.9±0.012 | Shall not exceed 5°c above the receiving water temperature |
| Rainy | 25.0±0.06 | 26.8±0.15 | 26.2±0.02 | ||
| Winter | 17.5±0.03 | 15.6±0.07 | 17.6±0.10 | ||
| pH | Summer | 8.5±0.06 | 6.9±0.03 | 7.0±0.09 | 5.5-8.5mg/L |
| Rainy | 8.0±0.06 | 7.0±0.03 | 8.0±0.06 | ||
| Winter | 7.6±0.10 | 7.4±0.07 | 7.0±0.13 | ||
| DO mg/L | Summer | 7.3±0.02 | 8.5±0.01 | 10.5±0.02 | 30mg/L |
| Rainy | 9.8±0.02 | 10.2±0.01 | 10.3±0.06 | ||
| Winter | 8.3±0.03 | 10.3±0.03 | 10.4±0.03 | ||
| BOD mg/L | Summer | 473±1.722 | 277±2.314 | 76.8±0.983 | 30mg/L |
| Rainy | 482±1.453 | 288±2.887 | 77.5±0.345 | ||
| Winter | 501±1.145 | 235±1.577 | 72.1±1.201 | ||
| COD mg/L | Summer | 1126.6±1.146 | 430.8±1.640 | 425±2.145 | 250mg/L |
| Rainy | 1180±2.886 | 681±7.264 | 291±4.509 | ||
| Winter | 1210±2.856 | 456±4.409 | 254±1.527 |
| Items | Season | Fish reared in cage in unpolluted water body | Fish caught from polluted Ramgarh lake | ||
|---|---|---|---|---|---|
| Site-1 | Site-2 | Site-3 | |||
| PLT(lac/mm3) | Summer | 188.7±0.88 | 20.7±0.64(11) | 25.2±0.09(13) | 32.2±0.12(17) |
| Rainy | 196.2±0.09 | 18.8±0.04(9) | 25.1±0.07(13) | 44.3±0.09(22) | |
| Winter | 176.6±0.45 | 18.3±0.003(10) | 20.4±0.15(11) | 21.6±0.009(12) | |
| WBC(103/mm3) | Summer | 5.6±0.06 | 0.3±0.06(5) | 0.6±0.25(11) | 0.4±0.01(17) |
| Rainy | 6.3±0.06 | 7.3±0.12(116) | 18.4±0.09(292) | 21.2±0.09(336) | |
| Winter | 7.30±0.03 | 1.29±0.08(17) | 1.36±0.16(19)) | 2.02±0.003(28) | |
| RBC(106/mm3) | Summer | 4.5±0.02 | 0.`45±0.01(10) | 0.5±0.01(11) | 1.3±0.01(29) |
| Rainy | 4.8±0.02 | 2.7±0.06(16) | 1.2±0.01(25) | 1.3±0.01(27) | |
| Winter | 5.37±0.15 | 2.23±0.004(41) | 2.32±0.003(43) | 3.13±0.10(58) | |
| HGB(g/dl) | Summer | 10.2±0.12 | 1.670.04(16) | 2.2±0.06(21) | 2.3±0.06(22) |
| Rainy | 9.5±0.01 | 4.5±0.01(47) | 5.7±0.01(60) | 9.3±0.01(97) | |
| Winter | 12.3±0.31 | 1.16±0.004(8) | 1.21±0.006(10) | 1.81±0.02(15) | |
| HCT | Summer | 351.7±0.88 | 5.5±0.01(1) | 10.5±0.07(3) | 15.6±0.12(4) |
| Rainy | 357.0±1.53 | 13.5±0.08(4) | 28.3±0.09(8) | 17.6±0.04(5) | |
| Winter | 325.5±2.75 | 17.2±0.05(5) | 17.01±0.19(5) | 15.26±0.03(5) | |
| MCV(µg) | Summer | 54.7±0.88 | 12.2±0.06(22) | 13.4±0.01(24) | 15.5±0.02(28) |
| Rainy | 53.7±1.20 | 10.5±0.09(19) | 11.3±0.09(21) | 13.7±0.06(26) | |
| Winter | 60.26±0.05 | 14.5±0.006(24) | 13.35±0.005(22) | 14.57±0.14(24) | |
| MCH(pg) | Summer | 47.6±0.16 | 13.5±0.06(28) | 20.5±0.12(43) | 33.4±0.01(70) |
| Rainy | 44.0±0.09 | 17.6±0.01(40) | 35.2±0.12(80) | 37.5±0.15(85) | |
| Winter | 58.0±0.17 | 18.2±0.003(31) | 22.21±0.005(38) | 23.3±0.004(40) | |
| MCHC(g/dl) | Summer | 30.4±0.09 | 10.6±0.25(35) | 14.3±0.06(47) | 25.2±0.09(82) |
| Rainy | 39.3±0.07 | 33.2±0.12(84) | 32.7±0.06(83) | 36.4±0.37(92) | |
| Winter | 28.8±0.28 | 20.84±0.09(72) | 22.08±0.036(76) | 21.78±0.04(76) | |
| Parameters | Season | Tissue (50mg) | Fish reared in unpolluted water body | Magnitude | ||
|---|---|---|---|---|---|---|
| Site-1 | Site-2 | Site-3 | ||||
| Protein (µg/mg) | Summer | Muscle | 96.0±0.02 | 20.13.±0.07(27) | 30.96±0.005(32) | 42.28±0.015(44) |
| Liver | 75.3±0.01 | 22.55±0.025(30) | 25.06±0.009(33) | 45.45±0.21(60) | ||
| Rainy | Muscle | 89.2±0.02 | 30.31±0.008(34) | 31.22±0.03(35) | 43.12±0.02(48) | |
| Liver | 72.0±0.01 | 25.61±0.006(35) | 24.86±0.004(34) | 33.11±0.005(45) | ||
| Winter | Muscle | 60.2±0.004 | 25.45±0.02(42) | 23.56±0.05(39) | 27.76±0.03(46) | |
| Liver | 64.5±0.002 | 22.14±0.05(34) | 22.68±0.01(35) | 38.51±0.02(60) | ||
| Amino acid (µg/mg) | Summer | Muscle | 7.2±0.08 | 38.22±0.16(530) | 34.08±0.09(413) | 36.35±0.04(504) |
| Liver | 6.9±0.09 | 37.63±0.39(545) | 26.23±0.07(380) | 21.98±0.06(318) | ||
| Rainy | Muscle | 8.2±0.18 | 40.7±0.05(496) | 35.9±0.33(437) | 41.5±0.50(506) | |
| Liver | 7.8±0.02 | 38.7±0.10(496) | 35.3±0.03(455) | 39.9±0.35(532) | ||
| Winter | Muscle | 6.3±0.01 | 20.5±0.07(325) | 32.7±0.46(519) | 34.5±0.006(547) | |
| Liver | 6.5±0.02 | 28.5±0.002(538) | 30.3±0.042(466) | 27.2±0.009(418) | ||
| Glycogen (µg/mg) | Summer | Muscle | 14.0±0.01 | 9.09±0.01(64) | 9.37±0.08(67) | 9.47±0.007(67) |
| Liver | 17.2±0.02 | 8.40±0.007(48) | 9.47±0.006(55) | 9.46±0.01(55) | ||
| Rainy | Muscle | 12.15±0.85 | 7.61±0.009(67) | 8.93±0.16(73) | 9.20±0.01(74) | |
| Liver | 15.3±0.16 | 6.71±0.002(44) | 8.70±0.002(56) | 9.19±0.01(60) | ||
| Winter | Muscle | 10.6±0.009 | 8.03±0.008(75) | 9.1±0.07(86) | 9.3±0.04(93) | |
| Liver | 10.5±0.002 | 8.2±0.08(78) | 9.05±0.01(86) | 9.3±0.01(88) | ||
| DNA (µg/mg) | Summer | Muscle | 22.5±0.01 | 10.8±0.01(48) | 12.0±0.002(53) | 15.3±0.02(68) |
| Liver | 27.0±0.002 | 7.1±0.001(26) | 12.5±0.01(46) | 15.0±0.01(55) | ||
| Rainy | Muscle | 24.3±0.02 | 8.3±0.17(34) | 13.5±0.01(55) | 14.10.004(58) | |
| Liver | 28.0±0.001 | 9.4±0.005(33) | 10.2±0.15(36) | 12.4±0.09(44) | ||
| Winter | Muscle | 25.6±0.003 | 13.3±0.02(52) | 14.0±0.004(55) | 15.3±0.02(60) | |
| Liver | 25.8±0.009 | 12.3±0.001(48) | 12.0±0.01(46) | 10.5±0.002(47) | ||
| RNA(µg/ mg) | Summer | Muscle | 27.2±0.005 | 10.8±0.01(40) | 11.2±0.05(41) | 22.4±0.001(82) |
| Liver | 28.5±0.002 | 12.4±0.02(45) | 16.3±0.006(57) | 24.4±0.001(86) | ||
| Rainy | Muscle | 27.0±0.002 | 13.2±0.005(48) | 14.8±0.02(55) | 22.0±0.001(81) | |
| Liver | 30.5±0.001 | 11.6±0.01(35) | 15.3±0.02(50) | 25.5±0.02(83) | ||
| Winter | Muscle | 26.7±0.004 | 10.8±0.01(40) | 11.5±0.007(43) | 20.5±0.005(77) | |
| Liver | 28.5±0.002 | 12.3±0.001(43) | 13.2±0.005(46) | 15.05±0.01(53) | ||
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| How to cite this article: Mamta K, Singh A: Hematological and Biochemical Changes Induced by Water Pollutants in Fishes Collected from Ramgarh Lake of Gorakhpur (U.P) India. Int. J. Life. Sci. Scienti. Res., 2017; 3(1): 792-799. DOI:10.21276/ijlssr.2017.3.1.8 Source of Financial Support: Nil, Conflict of interest: Nil |