acute toxicity effect of imidacloprid insecticide on serum biochemical

B.Padma Priya et al ISSN 2278-1145

Int. J. Int sci. Inn. Tech. Sec. A, Jun. 2012, Vol. 1, Issue 2, pg18-22

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International Journal of Integrative sciences, Innovation and Technology

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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: [email protected]

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 (C9H10CIN5O2), 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 40C prior to

immediate determination of biochemical parameters,

Glucose, Protein, Cholesterol, Creatinine and

Creatine. Glucose was Folin-Wu6 method determined

by method, Protein, Albumin and Globulin

determined by Reinhold’s7 method, Cholesterol

determined by Zak’s8 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 Students‘t’ 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 47.33±0.74 51.54±.087 54.34±0.91 57.86±0.98 60.90±0.99 62.56±1.0

(59.06)* (59.36)* (58.90)* (61.34)* (62.16)*

Protein 11.37±0.57 9.34±0.45 8.5±0.41 8.27±0.33 7.56±0.31 7.41±0.30

(20.56)* (20.34)* (24.77)* (24.35)* (24.85)*

Albumin 1.61±0.061 1.33±0.058 1.09±0.054 0.85±0.047 0.74±0.040 0.61±0.035

(22.85)* (20.10)* (17.77)* (18.34)* (17.05)*

Globulin 8.40±0.37 8.25±0.194 7.78±0.092 7.34±0.082 6.65±0.08 6.36±.0.081

(42.36)* (84.18)* (88.96)* (82.56)* (78.36)*

Cholesterol 124±2.06 137.03±2.51 143.76±3.19 146.73±3.21 155.43±3.65 162.04±4.26

(54.50)* (45.06)* (45.88)* (42.54)* (38.02)*

Creatinine 0.54±0.015 0.61±0.017 0.67±0.020 0.75±0.22 0.77±0.021 0 .83±0.029

(34.43)* (33.39)* (33.32)* (35.48)* (28.30)*



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Creatine 0.55±0.015 0.60±0.018 0.67±0.018 0.72±0.026 0.79±0.031 0.99±.034

(33.20)* (35.98)* (26.94)* (25.23)* (25.62)*

Values are mean±SE 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

condition9. Moreover, analysis of serum biochemical

constituents’ levels showed useful information in

detection and diagnosis of metabolic disturbances

and diseases in fishes10

.

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 catecholamines14

. 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 stress27

. 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



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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acute toxicity effect of imidacloprid insecticide on serum biochemical

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