Indo American Journal of Pharmaceutical Research, 2014

ISSN NO: 2231-6876

SUB-LETHAL EFFECT OF CYPERMETHRIN ON DIFFERENT ADENYLATE

NUCLEOTIDES IN VARIOUS TISSUES OF CHANNA STRIATUS (BLOCH)
Md. Osman1 Ahmed, Rabia Banu1 and S.A. Mastan2
1

Department of Zoology, Osmania College UG & PG, (Autonomous) Kurnool-518 001, A.P, India
Matrix- ANU Advanced Aquaculture Research Centre, Acharya Nagarjuna University, (MAAARC), Nagarjuna Nagar-522 510, A .P.
India.
2

ARTICLE INFO
Article history
Received 31/12/2015
Available online
05/01/2015

Keywords
Cypermethrin,
Adenylate Nucleotides,
Channa Striatus.

ABSTRACT
The aim of the present study is to evaluate the effect of sub-lethal effect of cypermethrin on
different adenylate nucleotides in various tissues of Channa striatus. It has been observed
that the levels of ATP, ADP and energy charge elevated at 24 hours relative to control in all
the organs. But their levels declined through 7 and 15 days exposure periods. The percent of
suppression is high at 15 days exposure period. From 15 th day onwards their level gradually
elevated and came nearer to control at 30 day exposure period and the values were found to
be significant. In case of AMP it has been observed that AMP declined at 24 hours relative
to control in all the organs. But their levels elevated gradually through 7 day and continued
upto 15 days exposure period. From 15 th day onwards levels of AMP gradually declined and
came nearer to control at 30 day exposure period and the values were found to be significant
at (P< 0.001). Hence from the results of present study it can be concluded that cypermethrin
was shown significant toxic effect on adenylate nucleotides in various tissues of C. striatus

Copy right 2014 This is an Open Access article distributed under the terms of the Indo American journal of Pharmaceutical
Research, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Please cite this article in press as Dr. Md. Osman Ahmed et al. Sub-Lethal Effect of Cypermethrin on Different Adenylate
Nucleotides In Various Tissues of Channa striatus (Bloch. Indo American Journal of Pharm Research.2014:4(12).

5949

Corresponding author
Dr. Md. Osman Ahmed
Department of Zoology
Osmania College (Automous)
Kurnool-518 001
Andhra Pradesh, India

Vol 4, Issue 12, 2014.

Dr. Md. Osman Ahmed et al.

ISSN NO: 2231-6876

INTRODUCTION
Indiscriminate application of various pesticides has aggravated the problem of pollution to aquatic environment. Due to these
synthetic chemicals, environment has failed to keep its healthy nature. The insecticides of proven economic potentialities could not do
well in the ecosystem when viewed on extra fronts since these revenue poisons, in a residual form or as a whole, get into the aquatic
ecosystem. They cause a series of problems to aquatic organisms, especially for the fishes [33, 23, 26]. Pesticides have a great impact
on human health, production and preservation of food, fishes and other cash crops. On the other hand use of pesticides has given rise
to many serious problems. Most of the chlorinated non-degradable pesticides leave residues in various living systems for prolonged
periods of their life span and are presumably responsible for a variety of known and unknown toxic symptoms. Even when present in
minute quantities, their variety, toxicity and persistence has an adverse effect on ecological systems, such as birds, fishes and plants,
with which human welfare is inseparably bound [25, 32]. Cypermethrin, a synthetic pyrethyroid, is broad spectrum insecticide,
used extensively in households, industrial and agriculture fields, for control of several pests [37]. Due to indiscriminate use of
cypermethrin makes their entrance into natural water bodies through agricultural runoff and ultimately affect the aquatic organisms,
adversely affect fish metabolism [22, 28].
The most important adenylates that are available in the animal body are ATP, ADP and AMP. The energy relationships of
these different nucleotides are quite different from one another. The hydrolysis of the diphosphate bond and triphosphate bonds are
associated with the release of large amount of free energy approximately 8000 cal/mole, while the phosphate bond of adenyclic acid is
associated with only 1000 or 2000 cal/mole. Therefore former are referred to as high energy rich compounds while latter are called
low energy nucleotides. Most of the specific tasks of the animal such as concentration or secretion depend directly on the ATP, ADP
phosphate transfer system and for some biosynthetic processes; the phosphate bond energy comes from the hydrolysis of ADP to form
AMP. In fact the energy status of the cell depends on the relative amounts of those three derivatives. In many important metabolic
reactions these phosphate compound are considered as the primary source of energy and serves as allosteric modulators. A few studies
involving the adenylate nucleotides and energy charge with reference to environmental pollution. Further, these are confined to
chlorinated hydrocarbons that too mostly confined to insects and mammals. The first research on ATP showed no adverse affect
resulting from DDT toxicity). Although in later studies the inhibition of oxidative phosphorylation was demonstrated by using 100
moles of DDT. However, in 1969 the first report on inhibition of ATPase by organ chloride pesticide was made by [18, 36, 26 ]. This
paper communicates the sub-lethal effect of cypermethrin on different adenylate nucleotides in various tissues of fish, C. striatus

Energy Charge
The adenylate energy charge (A.E.C) values were calculated by the following formula as given by the Atkinson (1968).
AEC = ATP +1/2 ADP
ATP + ADP + AMP

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Technical grade cypermethrin (95%) was obtained from United Phosphorus Ltd., Mumbai, India.
The concentration of adenylate nucleotides like ATP, ADP and AMP are estimated separately in the organs like gill, kidney,
intestine, brain, liver and muscle of C. striatus at different sub-lethal exposures of cypermethrin. The similar estimations were also
done in tissues of the fish in freshwater without cypermethrin served as controls. Prior to the estimation to stop the movements of the
fish at once and thereby in order to keep the nucleotide concentration non changed the fishes were anaesthetized with methane
sulphonate (MS 222) and transferred to heptan (-90 0C) and there after the fish were put immediately in the liquid air. Thus the fish
were freezed to death. Then the deep freezed tissues like gill, kidney, intestine, brain, liver and muscle were separated and ground to a
fine powder separately in a mortar. Separate homogenates were prepared for these samples in ice cold 60% perchloric acid solution.
The entire process of isolation and preparation of the tissue samples was carried out in an ice chamber with the temperature
maintenance of 15 + 10C. The homogenate was centrifuged for 15 minutes and the PCA supernatant were neutralized to pH 7.4 with
KOH and the precipitated KCIO4 was centrifuged for 10 minutes in high speed centrifuge this extraction procedure results in 95%
adenylate recovery. From this supernatants the determination of adenylate nucleotides like ATP, ADP, and AMP was carried out
according to the method of Bergmayer [6]. The measures were carried out in spectrophotometer at a wave length of 360nm and the
ATP, ADP, AMP concentrations were expressed in moles / gm wet weight of the tissue.

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MATERIALS AND METHODS

Procurement of test fishes
Alive, health and diseases free C. striatus (110 in no. weight between 10- g and length ranges 7-8 cm ) was collected from
Department of fisheries, Kurnool, A.P., and brought to the laboratory. Fishes were acclimatized in the laboratory condition for one
week in plastic pool. The water of plastic pool was aerated daily for two hours and water in plastic pool was replaced thrice in a week.
The fish were fed daily with ground nut oil cake and frog muscles twice in a week. An experiment of the present study is conducted
in static waters suggested by Doudroff et al., [12] .The fish were starved for 24 prior to the start of experiment. The obtained Lc50
value was (Finney, 1971) 0.3802 ppm. /96 hrs. The sub lethal concentration was reported to be taken approximately one- fifth of the
LC50 value i. e 0.0760ppm [29]. The experimental fishes were separated into six batches of 10 fishes in each aquarium (15 L capacity),
containing 10 L water. First batch run as control without cypermethrin and remaining five batches were exposed to sub lethal doses of
cypermethrin for 24hrs, 7 day, 15 day, 20 day and 30 day period. Each experiment was conducted in triplicate. Control and exposed
fishes were sacrificed at the end of each time interval .Gills, kidney, intestine, brain, liver and muscles were isolated and immediately
transferred to deep freezer prior to analysis.

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RESULTS AND DISCUSSION

The results on the levels of energy rich nucleotides and energy charge in the organs like gill, kidney, intestine, brain, liver
and muscle of C. striatus at 1,7,15,20 and 30 days exposure to sub-lethal concentration of cypermethrin besides controls are presented
in (tables 1, 2 & 3). For comparison, the differences in the levels of each parameter said above obtained between the controls and
experimental were converted as percentages of the corresponding control and these percent values are also presented in the same
tables and plotted against exposure period in (fig. 1, 2, 3). The percent recovery in the levels is calculated in relation to the level in the
control medium which is fixed at 100%.
Adenocine triphosphate, Adenocine diphosphate and Energy charge
The results ATP, ADP and energy levels are presented in the (table1&2 fig. 1, 2,). It has been observed that the levels of
ATP, ADP and Energy charge elevated at 24 hours relative to control in all the organs. But their levels declined through 7 day and 15
days exposure periods. The percent suppression was maximum at 15 day exposure period. From 15th day onwards their level
gradually elevated and came nearer to control at 30 day exposure period and the values were found to be significant.
Table 1: ATP concentrations (mol/gm wet. wt) in different tissues of C. striatus in different sub-lethal exposure periods of
cypermethrin.
S.No.

Organ

Control

Exposure Period in Days

24 hr
7 Days
15 Days

20 Days
30 Days
Gill
0.81
1.48
0.34
0.63
0.84
0.94
Mean
0.08
0.10
0.08
0.12
0.10
0.14
S.D.
% Change
23.84
-16.72
38-43
29.53
20.99
% Recovery
49.00
t - Test
P < 0.001 P < 0.001 P < 0.001 P < 0.001 P < 0.001 P < 0.001
II
Kidney
Mean
1.06
1.95
0.12
50.00
0.06
0.42
S.D.
0.07
0.05
0.10
0.14
0.12
0.16
% Change
29.08
30.71
50.65
32.67
20.91
% Recovery
79.08
t - Test
P < 0.001 P < 0.001 P < 0.001 P < 0.001 P < 0.001 P < 0.001
III
Intestine
Mean
0.75
0.91
0.40
0.30
0.50
0.74
S.D.
0.01
0.02
0.01
0.03
0.04
0.01
% Change
8.64
18.90
24.32
13.50
2.16
% Recovery
97.29
t - Test
P < 0.001 P < 0.001 P < 0.001 P < 0.001 P < 0.001 P < 0.001
IV
Brain
Mean
2.82
3.96
2.08
1.02
1.56
1.78
S.D.
0.04
0.10
0.12
0.08
0.14
1.32
% Change
23.65
-15.32
-37.34
26.14
21.57
% Recovery
78.42
t - Test
P < 0.001 P < 0.001 P < 0.001 P < 0.001 P < 0.001 P < 0.001
V
Liver
Mean
1.28
1.72
0.70
0.16
0.43
0.54
S.D.
0.09
0.12
0.08
0.06
0.16
0.14
% Change
13.41
17.68
34.14
25.91
22.56
% Recovery
77.43
t - Test
P < 0.001 P < 0.001 P < 0.001 P < 0.001 P < 0.001 P < 0.001
VI
Muscle
% Change
0.25
12.85
47.85
45.78
42.85
% Change
0.25
12.85
47.85
45.78
42.85
% Recovery
57.14
t - Test
P < 0.001 P < 0.001 P < 0.001 P < 0.001 P < 0.001 P < 0.001
Each value is a Mean of six individual measurements. The percent change in ATP concentration is calculated in relation to
ATP concentration in control medium (fresh water without cypermethrin). The percent recovery in ATP concentration at 30-day
exposure period is calculated in relation to its level in control medium which is fixed at 100%
Wet. Wt: wet weight

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ISSN NO: 2231-6876

Table 2: ADP concentrations (mol/gm wet. wt) in different tissues of C. striatus in different sub-lethal exposure periods of
cypermethrin.
S.No.

Organ

Gill
Mean
S.D.
% Change
% Recovery
t - Test
Kidney
Mean
S.D.
% Change
% Recovery
t - Test
Intestine
Mean
S.D.
% Change
% Recovery
t - Test
Brain
Mean
S.D.
% Change
% Recovery

II

III

IV

t - Test
Liver

Exposure Period in Days

24 hr
7 Days
15 Days

20 Days

30 Days

0.12
1.22

0.44
1.55
26.22

0.76
0.96
21.33

0.47
0.67
41.80

0.62
0.82
32.78

P < 0.001

P < 0.001

P < 0.001

P < 0.001

P < 0.001

0.75
0.95
22.13
77.86
P < 0.001

0.34
0.08

0.75
0.06
28.47

0.08
0.09
24.30

0.51
0.03
50.69

0.78
0.07
31.94

P < 0.001

P < 0.001

P < 0.001

P < 0.001

P < 0.001

1.09
0.02

1.26
0.01
12.60

0.04
0.02
-45.37

0.55
0.01
-28.57

0.75
0.01
-20.16

P < 0.001

P < 0.001

P < 0.001

P < 0.001

P < 0.001

1.26
0.04

2.18
0.12
28.22

0.49
0.03
23.61

0.67
0.06
45.70

1.10
0.09
32.51

1.48
0.13
20.85
79.14

P < 0.001

P < 0.001

P<1

P < 0.001

P < 0.001

P < 0.001

Control

1.10
0.05
16.16
83.33
P < 0.001
1.04
0.02
15.40
94.80
P < 0.001

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5952

Mean
0.76
1.42
0.15
0.57
0.88
0.99
S.D.
0.06
0.08
0.18
0.07
0.04
0.09
% Change
23.91
22.10
42.75
31.52
25.36
% Recovery
74.63
t - Test
P < 0.001 P < 0.001 P < 0.001 P < 0.001 P < 0.001 P < 0.001
VI
Muscle
Mean
0.08
0.26
0.74
0.56
0.62
0.68
S.D.
0.09
0.13
0.07
0.12
0.15
0.09
% Change
15.25
20.33
35.59
30.50
25.42
% Recovery
75.57
t - Test
P < 0.001 P < 0.001 P < 0.001 P < 0.001 P < 0.001 P < 0.001
The percent change in ADP concentration is calculated in relation to ADP concentration in control medium (fresh water
without cypermethrin). The percent recovery in ADP concentration at 30-day exposure period is calculated in relation to its in the
control medium which is fixed at 100%.

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ISSN NO: 2231-6876

Fig. 1: The percent change over control in Adenocine tryphosphate (ATP) activity in different organs of C. striatus at
different day of exposure to sub-lethal concentration of cypermethrin

Gill
Kidney
Intestine

120.00

Brain

100.00

Liver

% Change

80.00

Muscle

60.00
40.00
20.00
0.00
-20.00

24 Hrs

7 Days

15 Days

20 Days

30 Days

% Recovery

-40.00
-60.00

Exposure Periods in days

Fig 2: The percent change over control in Adenocine diphosphate (ADP) activity in different organs of C. striatus at
different day of exposure to sub-lethal concentration of cypermethrin.

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5953

Adenocine monophoshate
The data presented in the table 3, and fig.4 5. It has been observed that the levels of AMP declined at 24 hours relative to
control in all the organs. But their levels elevated gradually through 7 day and continued upto 15 days exposure period. From 15 day
onwards levels of AMP gradually declined and came nearer to control at 30 day exposure period and the values were found to be
significant (P< 0.001).

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Table 3: AMP concentrations (mol/gm wet. wt) in different tissues of C. striatus in different sub-lethal exposure periods of
cypermethrin.
S.No.

Organ

Gill
Mean
S.D.
% Change
% Recovery
t - Test
Kidney
Mean
S.D.
% Change
% Recovery
t - Test
Intestine
Mean
S.D.
% Change
% Recovery
t - Test
Brain
Mean
S.D.
% Change
% Recovery
t - Test
Liver
Mean
S.D.
% Change
% Recovery
t - Test
Muscle
Mean
S.D.
% Change
% Recovery
t - Test

II

III

IV

VI

Exposure Period in Days

24 hr
7 Days

15 Days

20 Days

30 Days

0.70
0.09

0.52
0.18
-20.00

0.76
0.12
6.60

0.08
0.05
11.10

0.74
0.07
4.40

P < 0.001

P < 0.001

P < 0.001

P < 0.001

P < 0.001

0.62
0.14
8.80
91.11
P < 0.001

0.18
0.05

0.71
0.08
-28.90

0.17
0.10
7.81

0.30
0.07
9.30

0.10
0.12
6.25

P < 0.001

P < 0.001

P < 0.001

P < 0.001

P < 0.001

0.58
0.02

0.30
0.01
-41.17

0.60
0.01
2.94

0.69
0.01
16.17

0.62
0.01
5.88

P < 0.001

P < 0.001

P < 0.001

P < 0.001

P < 0.001

0.81
0.04

0.23
0.12
-26.37

0.84
0.06
7.14

1.20
0.14
26.37

0.95
0.09
12.63

P < 0.001

P < 0.001

P < 0.001

P < 0.001

P < 0.001

0.09
0.12

0.79
0.06
-25.00

0.09
0.14
8.33

0.25
0.11
12.50

0.14
0.09
3.30

P < 0.001

P < 0.001

P < 0.001

P < 0.001

P < 0.001

0.50
0.05

0.37
0.07
-18.57

0.46
0.08
5.71

0.64
0.10
20.00

0.55
0.13
7.14

P < 0.001

P < 0.001

P < 0.001

P < 0.001

P < 0.001

Control

0.06
0.16
9.37
90.62
P < 0.001
0.45
0.01
19.11
80.60
P < 0.001
0.61
0.07
6.04
93.96
P < 0.001
0.02
0.18
6.60
93.96
P < 0.001
0.46
0.11
11.42
88.57
P < 0.001

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The percent change in AMP concentration is calculated in relation to AMP concentration in control medium (fresh water
without cypermethrin). The percent recovery in AMP concentration at 30-day exposure period is calculated in relation to its in the
control medium which is fixed at 100%

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The variation in adenylate energy nucleotides concentration is correlated to the energy budget and hence energy demands of
the animal [8, 9]. Further, it is known that the primary energy source, the adenylate nucleotides like ATP, ADP and AMP serve as
allosteric modulators in many important metabolic reactions. Atkinson [2] refers to the ATP, ADP and AMP status of a cell as its
energy charge. ATPase is a complex enzyme system which catalyses the terminal step of oxidative phosphorylation and mediates
many ATP dependent activities including the hydrolysis of ATP. Even a slight decrease charge leads to many profound variations in
the concentration of many metabolites [2].
In the present study relative to controls and elevation in the levels of ATP, ADP and energy charge followed by decrease in
AMP were noticed in different organs of C. striatus on exposure to sub-lethal concentration of cypermethrin. Thus, the maximum
levels of ATP and energy charge in all the tissues of C.striatus during pesticide exposure reflects the greater activity of mitochondrial
electron transport system and hence the higher level of ATP turn over and energy (ATP) expenditure. Hence variations in energy rich
nucleotide like ATP, ADP and in the level of energy charge are ultimately correlated to the energy demands of the fish as also
suggested by Clark [8, 9].
An initial elevation at 24 hours exposure period in energy rich nucleotides ATP, ADP and energy charge and an increase in
oxygen consumption and in blood glucose level as noticed, it may be attributed to escape reaction which is due to increased
locomotors activity arising out of the animals tendency to escape from the stress medium [3, 1]. Further an increase in locomotary
activity is also very much seen in C. striatus during the initial period of 24 hours of cypermethrin exposure in the present
investigation.
It is known that as a primary energy source, the adenylate nucleotides like ATP, ADP and AMP serve as allosteric
modulatorts in many important metabolic reactions. Atkinson [2] refers to the ATP ADP AMP status of a cell as its energy. In
the arena of toxicology studies, it is also known that the level of the concentrations of these nucleotides is apparently decreased by the
insecticidal organochloride compounds [11, 15, 24] Therefore, the estimation of the levels of these adenylate nucleotides ATP, ADP,
AMP and energy charge available in different tissues of an animal namely a fish in this investigation could be taken as an indicator of
the extent of pollution of aquatic environment by the pesticides.
In most of the sub-lethal exposure periods in all the tissues, the ATP and ADP concentration decreased with an increase in
AMP concentration and the system was partially charged. Hence, the energy charge is correspondingly brought down; this decrease in
energy charge is maximally noticed in the 15th day exposure of cypermethrin. Thus, the energy charge involving availability of
different nucleotides, ATP, ADP, and AMP is one of the regulatory inputs to most and probably all metabolic sequences. Even a
slight decrease in energy charge leads to many profound variations in the concentration of many metabolites [2]. Such an effect is
also seen in the earlier studies where there is a significant decrease in energy reserves like liver glycogen and muscle glycogen in the
fish during most of the sub-lethal exposure periods of cypermethrin.
Further, there is an overall decrease in energy rich nucleotides like ATP, ADP and energy charge in all the tissues in the fish
during the sub-lethal exposure periods of cypermethrin. In this context, it is necessary to mention that ADH is a vital enzyme of TCA
cycle catalysing the dehydrogenase of succinate to fumarate while cytochrome oxidase is the key enzyme of terminal oxidative
system, reflecting the activity of mitochondrial electron transport system and hence ATP turn over as also reported by Rafi Ashamed
[30] in case of Labeo rohita during phosphomidon exposure, and therefore variations in the energy rich nucleotide like ATP are
ultimately correlated to the energy demand of the animal [8,9]. Thus whatever may be the tissue and whatever might be the level of
depletion in these nucleotides and energy charge, the reduction conspicuously in ATP, ADP and energy charge obviously reflects
impairment of oxidative metabolism at the mitochondrial level in the major carp, C. striatus exposed to sub-lethal concentration of
cypermethrin.

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Fig. 3: The percent change over control in Adenocine Monophaosphate (AMP) activity in different organs of C.
striatus at different day of exposure to sub-lethal concentration of cypermethrin.

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Moreover, these suppressed changes in the adenylate nucleotide system might be due to conditions similar to asphyxia
leading to inhibition of respiratory activity (oxygen consumption). Asphyxiation results in the reduced oxygen supply to the tissues
which in turn lead to decreased oxidation of substrates with the result that energy production is considerably reduced in this carp
exposed to cypermethrin. Thus inhibition of energy nucleotides like ATP, ADP and energy charge indicate a reduction in the
oxidation of succinate and probably due to progressive degeneration of mitochondria along with deformation. Therefore it is likely
that mitochondrial destruction and depressed oxidation of acetate could be one of the causes leading to diminished energy nucleotides
like ATP, ADP and energy charge in this carp exposed to cypermethrin. In fact, it is interesting to note that organophosphate
pesticides like sumithrion suppressed tissue respiration by inhibiting aerobic enzyme system in the fish, Tilapia mosambica [20].
Similarly, a decrease in the activity of SDH in some of the tissue of Tilapia mossambica exposed to sub-lethal concentration
of methyl parathion was also reported [34] and of Lamellidens marginalis exposed to [16]. Moreover, a number of pesticides were
shown to interfere with energy metabolism in fish [11] and hence it is possible that their main effect may be via the mitochondrial
energy metabolism, inhibiting the ATPase activity as also noticed in the present investigation in a variety of pesticides [11]. Therefore
it is possible that energy metabolism in the form of ATP production might have been reduced in C. striatus exposed to Deltamethrin
as relfected in their depleted ATP, ADP concentrations and energy charge levels.
The interesting feature of these findings is that in any tissue of fish, the energy rich nucleotides like ATP, ADP and the
energy charge does not consistently decrease with increase in time of sub-lethal exposure of cypermethrin. Thus, relative to control
medium (freshwater without cypermethrin the said parameters in all the tissues increased at 24 hours exposure period and registered a
steep decrease through 7th day period and this decrease was maximal at the 15 th day period of exposure ( tables- 1,2 fig- 1,2,3& 4).
Later, from its decrease at 15th day exposure period there is an increase in the levels of these ATP, ADP and energy charge in all the
tissues during 30th day exposure periods and these levels reached nearer to the levels of the control medium ( freshwater without
cypermethrin).
Until 15thday exposure period the activation of detoxification mechanisms concerned with cypermethrin toxicity might not
have been brought about as reflected in the depletion of these energy rich nucleotides. Hence, there is no filling up processes of ATP
as noticed in the lowest energy charge of the tissues during 15 th day exposure. Thus ATP, ADP and AMP are profoundly affected due
to cypermethrin stress. A similar inhibition on the energetic but with the organ chloride pesticides in fish population supports the
present findings, in that the ATPase system in fish tissue homogenates, has been evaluated with certain cyclodien insecticides and
related chemicals. These studies have been shown to have prominent inhibitory effects on Na+, K+ ATPase, oligomycin, Mg2+ATPase [11] Further the inhibition of oligomycin sensitive Mg 2 ATPase in liver paralled the impairment of hepatobiliary function
[11] suggesting that chlordane may impair the energy metabolism by interfering with OS Mg2+ ATPase in mitochondria.
However, at the end of 30th day sub-lethal exposure period of cypermethrin the adenylate nucleotides like ATP, ADP and
AMP and energy charge in all the tissues of fish exhibited a fairly good amount of recovery in their levels from earlier suppression at
15th day exposure period. It is significant to state that the capacity of a fish population to compensate for the effect of a pesticide was
also suggested by Coppage and Duke [7] where the AChE activity of the fish brain returned to normal within 40 days after application
of malathion. Such compensatory mechanism in the form of recovery during cypermethrin toxicity towards the end of the sub-lethal
exposure and such detoxifying enzyme enhancement has been reported [25] where it was found that an increase in the activity of
detoxifying enzyme like cytochrome, the terminal oxidize of the microsomal mixed function oxygenase (MFO) system to reduce
malathion toxicity in the liver and gill of the catfish. Further, this fairly good amount of recovery in all these nucleotide
concentrations and energy charge is a reflection (during the 30t h day period) of filling up processes with the ATP as noticed in the
regarding of the energy charge. Similar filling up processes with ATP and regarding of energy charge was noticed in the brain and
muscle tissues of the fish Idus idus during temperature adaptation process [19, 20, 21, 31]. Ghosh [14] studied the influence of
cypermethrin on oxidative metabolism of fish, Labeo rohita.

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CONCLUSION
In most of the sub-lethal exposure periods in all the tissues, the ATP and ADP concentration decreased with an increase in
AMP concentration and the system was partially charged. Hence, the energy charge is correspondingly brought down; this decrease in
energy charge is maximally noticed in the 15th day exposure of cypermethrin. Thus, the energy charge involving availability of
different nucleotides, ATP, ADP, and AMP is one of the regulatory inputs to most and probably all metabolic sequences. Even a
slight decrease in energy charge leads to many profound variations in the concentration of many metabolites. Such an effect is also
seen in the earlier studies where there is a significant decrease in energy reserves like liver glycogen and muscle glycogen in the fish
during most of the sub-lethal exposure periods of cypermethrin. In the present study it has been observed that cypermethrin has
significant toxic effect on different durations of its exposure on ATP, ADP and AMP in the different tissues of experimental fishes.
This kind study will be required on other species of food fishes.

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