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International Journal of Integrative sciences, Innovation and Technology
(A Peer Review E-3 Journal of Science Innovation Technology)
Section A Basic Sciences; Section B Applied and Technological Sciences; Section C Allied Sciences Available
online at www.ijiit.webs.com
Research Article
ACUTE TOXICITY EFFECT OF IMIDACLOPRID INSECTICIDE ON SERUM BIOCHEMICAL
PARAMETERS OF FRESH WATER TELEOST CHANNA PUNCTATUS
B.PADMA PRIYA*, VIJAYA RACHEL AND Y.AVASN MARUTHI

Dept. of Environmental Science & Dept. of Biochemistry , GITAM Institute of Science
GITAM University, Visakhapatnam, Andhra Pradesh
Corresponding Author Email address: padmapriyabu@ymail.com


ABSTRACT

The present study was designed to assess the effect of chloro-nicotinyl insecticide Imidacloprid on serum
biochemical parameters of fresh water teleost Channa punctatus. The test fish exposed to different concentrations of
toxicant to study the biochemical indices Glucose, Protein, Albumin, Globulin, Cholestrol, Creatinine and Creatine.
The Imidacloprid effect in test fish was studied after 96 h based on the results of acute toxicity tests and comparison
of control with experimental groups. After 96 h the serum Glucose (47.33-62.56), Cholestrol (124-162.04),
Creatinine (0.54 -0.83) and Creatine (0.55 - 0.89) have been increased significantly. On the other hand the serum
Protein (11.37-7.41), Albumin (1.61-0.613) and Globulin(8.40-6.36) have been decreased significantly after
Imidacloprid treatment with different concentrations (0.002ppm, 0.004ppm,0.006ppm, 0.008ppm and 0.010ppm).
The alterations of the above serum biochemical parameters could be used as an important tool to assess the
physiological and metabolic activity of test fish.

KEYWORDS: Imidacloprid, toxicity, biochemical, Channa punctatus, Serum

INTRODUCTION

Environmental pollution is a worldwide problem in a
modern society. The extensive use of pesticides are
widely used to enhance the crop production and other
benefits and has raised concerns about potential
adverse effects on the environment, human health
and non-target animals. Unfortunately, the
application of these derivatives of pesticides are
highly toxic to a number of non-target organisms
such as bees, fresh water fishes and other aquatic
organisms even at very low concentration
1-3
. Fishes
are one of the most widely distributed organisms in
an aquatic environment and being susceptible to
environmental contamination may reflect the extent
of the biological effects of environmental pollution in
waters. Monitoring of blood parameters, both
cellular and non-cellular may have considerable
diagnostic value in assessing early warning signs of
pesticide poisioning
4
.

Blood is a pathophysiological reflector of the whole
body. Knowledge on the physiological action of the
toxicant helps to predict an important sub lethal
effects and analysis of biochemistry, hematology and
histopathology may be used to determine the mode of
action of the toxicant. In recent years, biochemical
variables were used more when clinical diagnosis of
fish physiology was applied to determine the effects
of external stressors and toxic substances. Therefore,
the biochemical evaluations are gradually becoming a
routine practice for determining the health status in
fish. The biochemical parameters like Serum
Glucose, Protein, Cholesterol, Creatinine and
Creatine are widely used to determine the toxic stress
5
.

Imidacloprid is a systemic chloro-nicitnyl insecticide.
Its original name is Imidacloprid. It is a patented
chemical manufactured by Bayer Cropscience (part
of Bayer AG) and sold under trade names, Kohinoor,
Admire, Advantage, Merit, Imidacloprid and
Premise. It was the first member of a new family, the
neonicotinoids, and is chemically related to the
nicotinic acetylcholine receptor agaonists nicotine
and epibatidine Imidacloprid is a Chloronicotinyl
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19
insecticide, 1-((6-Chloro-3-pyridinyl)methyl)-N-
nitro-2-imidazoldinimine (C
9
H
10
CIN
5
O
2
), is a
nicotine mimic (nicotinoid insecticide) that belongs
to a group of insecticides referred to as the
choloronicotinyl group. Only a few studies were
identified the toxicity of common transformation
products of Imidacloprid to invertebrates.

MATERIALS AND METHODS

The adult Channa punctatus weighing 20-25cm
have been chosen as the experimental fish to
evaluate the toxicity of Imidacloprid a systemic
insecticide widely used for agricultural applications
in various sectors. The fishes were procured from
River Sharada, Anakapalli, Visakhapatnam Dist.,
Andhra Pradesh, India. They were brought to the
laboratory and acclimatized under laboratory
conditions for a period of 2 weeks. The tubs were
disinfected with 0.01% KMnO4 solution and washed
thoroughly to prevent dermal infection and fishes
were maintained in circular plastic tubs with
dechlorinated water which was continuously aerated.
The fishes were fed once a day with commercial
pellets and rice bran. Water was renewed daily and
feeding to fishes was withheld 24 hrs before the
commencement of the experiment. Five groups of 10
acclimatized fishes were taken in each circular plastic
tub of 10 liters of water capacity and different
concentrations of Imidacloprid (0.002ppm, 0.00ppm,
0.006ppm, 0.008ppm and 0.01ppm) were added.
Respected controls were maintained simultaneously.
All experiments were carried out in triplicates.

At the end of exposure period, the control and
experimental fish were starved for 96hrs for
analyzing the biochemical assays. The blood samples
were taken immediately by caudal puncture method
using non-heparinized tubes. The blood was
immediately centrifuged at 1500rpm for 10 min.
Serum then removed and stored at 4
0
C prior to
immediate determination of biochemical parameters,
Glucose, Protein, Cholesterol, Creatinine and
Creatine. Glucose was Folin-Wu
6
method determined
by method, Protein, Albumin and Globulin
determined by Reinholds
7
method, Cholesterol
determined by Zaks
8
method and Creatinine and
Creatine method determined by using commercial
kits. The significance of sample means between
control and Imidacloprid treated fish was tested by
using Studentst Test.

RESULTS

Acute toxicity of fish Channa punctatus, exposed to
various concentrations of Imidacloprid showed many
biochemical alterations are presented in Table 1. The
mean values have been taken for discussion. Glucose,
Cholesterol, Creatinine and Creatine were increased
significantly (p<0.001) when compared to control
groups. On the other hand the levels of Protein,
Albumin and Globulin levels were decreased
significantly (p<0.001) when compared to control
group.

Table 1. Changes in the levels of Serum Biochemical Parameters in a fresh water fish Channa punctatus
exposed to Imidacloprid insecticide.
______________________________________________________________________________
Parameters Control 0.002ppm 0.004ppm 0.006ppm 0.008ppm 0.010ppm

Glucose 47.330.74 51.54.087 54.340.91 57.860.98 60.900.99 62.561.0
(59.06)* (59.36)* (58.90)* (61.34)* (62.16)*

Protein 11.370.57 9.340.45 8.50.41 8.270.33 7.560.31 7.410.30
(20.56)* (20.34)* (24.77)* (24.35)* (24.85)*

Albumin 1.610.061 1.330.058 1.090.054 0.850.047 0.740.040 0.610.035
(22.85)* (20.10)* (17.77)* (18.34)* (17.05)*

Globulin 8.400.37 8.250.194 7.780.092 7.340.082 6.650.08 6.36.0.081
(42.36)* (84.18)* (88.96)* (82.56)* (78.36)*

Cholesterol 1242.06 137.032.51 143.763.19 146.733.21 155.433.65 162.044.26
(54.50)* (45.06)* (45.88)* (42.54)* (38.02)*

Creatinine 0.540.015 0.610.017 0.670.020 0.750.22 0.770.021 0.830.029
(34.43)* (33.39)* (33.32)* (35.48)* (28.30)*
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Creatine 0.550.015 0.600.018 0.670.018 0.720.026 0.790.031 0.99.034
(33.20)* (35.98)* (26.94)* (25.23)* (25.62)*
Values are meanSE and in parenthesis () t test of seven individual observations. Values are significant at
p<0.0001

DISCUSSION

Biochemical analysis can provide valuable
information for monitoring the health conditions of
fishes. Biochemical changes depend on the fish
species age, the cycle of sexual maturity and health
condition
9
. Moreover, analysis of serum biochemical
constituents levels showed useful information in
detection and diagnosis of metabolic disturbances
and diseases in fishes
10
.

Determination of glucose concentration in blood
serum is widely used as an indication of stress
response. Generally, glucose is continuously
required as an energy source by all body cells and
must be maintained at adequate levels in the plasma
11
. In many fish species, the blood glucose level has
the tendency to increase due to experimental stress.
In the present study the significant increase in
glucose which was time dependent may be
considered to be manifestation of stress induced by
Imidacloprid insecticide. Wedemeyer et al. (1981)
12

stated that high levels of blood glucose are caused by
disorders in carbohydrate metabolism appearing in
the condition of physical and chemical stresses. A
variety of stressors stimulate the adrenal tissue
resulting in increased level of circulating
glucocorticoids
13
and catecholamines
14
. Both of
these groups of hormones produce hyperglycemia. It
is generally thought that, under conditions of stress,
hyperglycemia may provide additional energy during
times of high metabolic need such as fight and
flight response
15
. M.Ramesh and M. Sarvanan
(2008)
16
found similar results in Channa punctatus
when exposed to Chloropyrifos. E.Ogueji Okechuk
Wu and J. Auta (2007)
17
were also observed similar
results in Clarias garipinus, exposed to Lambda-
cylathrin

Proteins are mainly involved in the architecture of
the cell, during stress conditions of fish need more
energy to detoxify the toxicant to overcome stress.
The depletion of protein fraction in serum in present
study may have been due to their degradation and
possible utilization of degraded products for
metabolic purposes
18.
Decreased protein level may
be attributed to stress mediated immobilization of
these compounds to fulfil an increased element for
energy by the toxicant
19
. The depletion in total
protein content may be due to augumented
proteolysis and possible utilization of their product
for metabolic purposes
20
. The quantity of protein is
dependent on the rate of protein synthesis or on rate
of its degradation. The quality of protein may also be
affected due to impaired incorporation of amino acids
in polypeptide chains. Hussain et al., (1996)
21
reported that the decrease of total protein in Atrazine
(herbicide) treated fish Oreochromis niloticus and
Chrysichthyes auratus was mainly due to globulin.
Serum albumin measures as considerable diagnostic
value in laboratory animals because it relates general
nutritional status, the integrity of the vascular system
and liver function. Albumin in fish organism
pacrticipate in plastic metabolism and perform
transport functions of substances necessary for life
activities (first of all lipids)
22
. In our study Protein,
albumin and globulin were decreased significantly.
Lipika patnaik (2010)
23
also observed same results in
Sevin exposed to Clarias battrachus. Therefore, our
results were coinciding with the latest once.

Sterols are regarded as essential nutrients because if
the role cholesterol as a cell constituent and as a
precursor of steroid and moulitng hormones
24
.

The
liver plays a major role in Cholesterol homeostasis by
regulating plasma lipoprotein metabolism and lipid
output in bile
25-26
Cholesetrol in the blood is linked
to lipid metabolism and depends on the calorific
value of the feed
17
. Cholesterol is known to
participate in the rise of total lipid. The rise of these
energy reserves in response to pollution could be due
to the fact that excess energy reserves that as glucose,
triglycerides and cholesterol are required by
organisms to mediate the effects of stress
27
. In
addition, the abnormal accumulation of fats in
experimental animals could be due to induced
imbalance between fat production and utilization
28
.
Anand Pratap singh (2010)
18
studied that the decline
in Cholesterol level is due to utilization of stored and
circulatory cholesterol and other lipid fractions in
phorate treated fish countered toxic effect produced
and further stabilization of the toxic phorate
molecules to prevent harm caused by them.

Creatinine is derived mainly from the catabolism of
creatine found in muscle tissue and its catabolism to
creatinine occurs at a steady rate. Severe kidney
damage will lead to increased creatinine levels
29.
Well et al. (1986)
30
addressed that the raised serum
creatinine is indication of decreased glommerular
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21
filteration rate of fish posterior kidney. In the present
study serum Creatinine and Creatine showed
marginal increase in experimental group in
comparison to control animals and increase relates to
renal failure. A.Y Gaafar (2010)
31
observed that the
significant increase of creatinine in Oreochromis
niloticus following chronic exposure to edfenphos
pesticide.

It was concluded that Imidacloprid (chloro-nicotinyl)
insecticide has a profound influence on serum
biochemical profiles of test fish. Our results were
also consistent with previous researchers. Hence the
current results are helpful reference for evaluating the
physiological status and metabolic activity levels of
Imidacloprid treated fish.

ACKNOWLEDGEMENTS

The authors wish to thank the GITAM University
management for their kind encouragement and
laboratory facilities. The first author thankful to Dr.
I.Bhaskar Reddy, HOD, Dept.of Biochemistry GIS,
GITAM University for providing necessary
laboratory facilities and their constant guidance to
carry out the research work.

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