Toxicity of Polytrin-c on the psychopharmacological observations in wistar rats

Abstract

The work has been designed to asses the neuro toxicity of a combination pesticide through behavioral parameters [Psychopharmacological observation] and instrumental analysis such as spontaneous motor activity for 28 days. The mixed formulation selected for the study, polytrin-C was used for sub acute and acute toxicity studies on Wister rats weighing around 90-120 gms of either sex.. Acute studies were carried out to fix the LD 50 dosage of the selected pesticide. Based on the LD 50 values the sub acute doses were fixed [1/40th, 1/20th, 1/10th of the LD 50 Dosage]. It is evident from the results that 1/20 and 1/10 of LD 50 dosed animals of either sex showed marked differences in the behavioral patterns when compare to 1/40 of LD 50 dose animals. Due to the synergetic effect neurotoxicity symptoms of combination pesticide results show a different toxicological profile when compare with the toxicity of constituent pesticide.

Full text

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International Journal of Pharma and Bio Sciences V1(2)2010
TOXICITY OF POLYTRIN-C ON THE PSYCHOPHARMACOLOGICAL
OBSERVATIONS IN WISTAR RAT.

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SEKAR BABU HARI RAM*1, CH.UMADEVI1, GOWRI REDDY1,
T.M.VIJAYAKUMAR2 AND G.THIRUMURUGAN2.
1. Godavari Institute of Engineering and Technology, NH-5, Chaitanya nagar, Rajahmuundry-533294.
2. Research lab, GIET School of pharmacy, NH-5, Chaitanya nagar, Rajahmuundry-533294.

*For correspondence drhsekarbabu@yahoo.com,thiru.mpharm@gmail.com

ABSTRACT

The work has been designed to asses the neuro toxicity of a combination pesticide through
behavioral parameters [Psychopharmacological observation] and instrumental analysis such as spontaneous
motor activity for 28 days. The mixed formulation selected for the study, polytrin-C was used for sub acute
and acute toxicity studies on Wister rats weighing around 90-120 gms of either sex.. Acute studies were
carried out to fix the LD 50 dosage of the selected pesticide. Based on the LD 50 values the sub acute doses
were fixed [1/40th, 1/20th, 1/10th of the LD 50 Dosage]. It is evident from the results that 1/20 and 1/10 of LD
50 dosed animals of either sex showed marked differences in the behavioral patterns when compare to 1/40
of LD 50 dose animals. Due to the synergetic effect neurotoxicity symptoms of combination pesticide results
show a different toxicological profile when compare with the toxicity of constituent pesticide.

KEY WORDS

Neurotoxicity, Psychopharmacology, polytrin-C, Pesticides

INTRODUCTION

Man lives in the world of chemicals.
Chemicals are produced in widespread commercial
usage particularly as pesticides, medicines and
industrial chemicals. Although these chemical are
generally beneficial they have their negative aspects
too. Many of these chemicals may pollute the
environment and some eventually find their way to
humans through contaminated food, water and air. It
is now established that organophorous and
carbamates are toxic because of their interference in
the normal synaptic transmission of nerve impulse.
The Ops after entering the body of an organism
reaches the cholinergic sites of the nervous system
and inhibit the activity of the AChE by binding at its
active sites. AchE inhibition, thus leads to the
accumulation of Ach at nerve endings which in turn
cause the disruption of the nervous activity resulting
in excitation, paralysis and finally the death of the
organism.
Nowadays organophosphate
widely used as pesticide and have substantially
replaced many of more persistent chlorinated
esters are

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International Journal of Pharma and Bio Sciences V1(2)2010
TOXICITY OF POLYTRIN-C ON THE PSYCHOPHARMACOLOGICAL
OBSERVATIONS IN WISTAR RAT.

hydrocarbon insecticide1. Recently natural
pyrethroids have replaced organophosphates because
of their low mammalian toxicity and lack of
persistence in the environment2,3. Neuro toxicity is
one of the major effects induced by the organo-
phosphates4 and pyrethroids5,6, since lot of data
available regarding neuro-toxicity of constituent
pesticide [organophosphates or pyrethroids] ,but less
information available regarding the toxicity of
combination pesticide. It has been stated that
organophorous pesticides like chlorpyrifos and
profenos are compatible with pyrethroids like
cypermethrin2. Data on particular combination of
insecticides are scanty and general principles are
necessary for predictive purposes. The possible
types of toxicological interaction between two
insecticides are (1) additive, (2) synergistic and (3)
antagonistic or they may act independently of one
another. Often, compounds of same toxic effects act
additively which in case of insecticides with
different toxic effects; the combined effect is less
than additive. It has been shown that some pairs of
Ops exhibit greater than additive toxic effects when
administered together. Data on neurotoxicity of
combination pesticides are not available and it is
Compound Name: Polytrin C
Details of Groups and Dosage
Group I: Control [only distilled water]
Group II: 75-mgs/kg body-weight
Group III: 150 mgs/kg-body weight
Group IV: 300-mgs/kg-body weight
The control group [G1] received distilled water by oral intubations [single administration] at a dose of
10ml/kg body weight. Groups II, III and IV received different doses by oral gavages, [single administration]
of the pesticide dissolved in distilled water. The animals were observed 14 days after dosing and basing on
the mortality, the LD50 was calculated using Finney’s probit analysis9.



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only logical to believe that the neurotoxicity of
combination pesticides may differ from that of
individual pesticides. hence the present work has
been designed for 28 days sub acute studies to asses
the neuro toxicity of a combination pesticide through
psychopharmacological observation7.

MATERIALS AND METHODS

Wister rats [either sex]weighing around 90-
120 gms selected for the present study and they were
acclimatized and fed ad libitum with standard rat
pellets obtained from Lipton India Ltd Bangalore.
The new combination pesticide polytrin –C [organo
phosphate profenofos
cypermethrin 4% ] obtained from Aventis Bombay.
LD 50 studies [Acute Toxicity]
Acute Toxicity studies were carried out using forty
rats was randomly divided into four groups, each
group consisting of 10 animals [five females and
five males]. The allocation of the groups and
fixation of doses of the combination pesticides8 in
the different groups were carried as given below.
40% + pyretheroids

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TOXICITY OF POLYTRIN-C ON THE PSYCHOPHARMACOLOGICAL
OBSERVATIONS IN WISTAR RAT.

Sub acute Studies

Psychopharmacological studies reveal behavioral changes observed depending biochemical and
physiological changes at various morphological sites within the body due to the pesticide toxicity. Similar to
acute studies, sub acute psychopharmacological sub acute studies [28 days] were carried out by randomly
dividing the animals into four groups and each group consisting of 5 males and 5 females. Group I the
untreated control receiving only distilled water at a dose of 10 ml/kg body weight. The doses of the
combination pesticide in the sub acute studies were based on the LD 50 values obtained from acute studies.
Group II received 1/40th, Group III-1/20th, Group IV-1/10th of the LD 50 Dosage.

The allocation of group and doses were fixed as follows

Group I Control [distilled water]
Group II 8.5 mg/kg body weight
Group III 17 mg/kg body weight
Group IV 34 mg/kg body weight

The above mentioned dosages of the compounds were dissolved in distilled water and administrated
by oral gavages once daily for 28 days. During the period of study in-life parameters like body weight and
feed consumption were recorded. The psychopharmacological parameters to assess the Neuro toxicity of the
combination pesticide evaluated during the live phase of the sub acute studies10. Hence
psychopharmacological symptoms such as restlessness, convulsions and skeletal movements like ataxia,
catalepsy, gripping strength and righting reflex were studied in the compound treated with reference to the
respective untreated control11. The above observations were made on the 7th, 14th, 21st & 18th days using the
Actophotometer [Inco] and Digital Rotorod apparatus12.

RESULTS
In the 28-day sub acute study with polytrin-C
[profenos 40% + cypermethrin 4% ] in Wister rats, a
decrease in feed consumption and a related decrease
in body weight13,14 compared to control was
observed from the third week of treatment. Only
mild behavioral changes were observed and severe
signs of neurotoxicity like tremors and convulsions
were not observed in the polytrin-C treated groups
[TABLE 1 & 2] indicating polytrin-C to be less
neurotoxic combination pesticide because in the
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study polytrin-C treated rats did not show any
significant change in motor activity and muscle
relaxant property exhibited in female animals of
highest dose (34 mg/kg b.w.) Psychopharmalogical
parameters reflect the magnitude of neutoxicity and
in the present study, an incidence of the behavioral
changes was observed from the third week in rats
treated with Polytrin-C did not induce any abnormal
behavioral changes in male rats till the third week of
treatment and the fourth week in female rats
Psychopharmacological investigation revealed only
minor behavioral abnormalities in rats treated with

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International Journal of Pharma and Bio Sciences V1(2)2010
TOXICITY OF POLYTRIN-C ON THE PSYCHOPHARMACOLOGICAL
OBSERVATIONS IN WISTAR RAT.

poytrin-C at the doses of 17 and 34 mg/kg b.w.,
from the third week of the study. A decrease in
spontaneous motor activity [table 3] and muscle
relaxant property [table 4] was observed mainly in

Table 1
Behavioral parameters of rats treated with different concentration of
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the highest dose group (34 mg/kg b.w.) while only
minor inconsistent (not time related) changes were
observed in other groups.
Polytrin-C.

Groups Weeks
Behavioral Parameters
Alertness Passive/Active Grooming Aggressiveness Restlessness Tremors
M F M F M F
+ + A A + +
+ + A A + +
+ + A A + +
+ + A A + +
+ + A A + +
+ + A A + +
+ + A A + +
+ + A A + +
+ + A A + +
+ + A A + +
+ + A A + +
+ + P P + +
+ + A A + +
+ + A A + +
- - A P + +
+ - P P + -

M: Male A: Active +: Presence of activity N: Normal F: Female
P: Passive -: Absence of activity


M
N
N
N
N
N
N
N
N
N
N
N
-
N
N
N
N
F
N
N
N
N
N
N
N
N
N
N
N
-
N
N
N
N
M
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
F
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
-
M
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
F
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
I
01
02
03
04
01
02
03
04
01
02
03
04
01
02
03
04
II
III
IV

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TOXICITY OF POLYTRIN-C ON THE PSYCHOPHARMACOLOGICAL
OBSERVATIONS IN WISTAR RAT.

Table 2
Behavioral parameters of rats treated with different concentration of Polytrin-C.
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Groups Weeks
Behavioral Parameters
Convu
lsions
M
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
Ata
xia
Catalepsy
Gripping
Strength
M
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
Righting
Reflex
M
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
F M F
- -
- -
- -
- -
- -
- -
- -
- -
- -
- -
- -
- -
- -
- -
- -
- +
M
-
-

-
-
-

-
-
-
-
-

-
-
-
F
-

-
-
-
-
-
-

-
-
-
-
-
-
+
F
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
F
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
-
I
01
02
03
04
01
02
03
04
01
02
03
04
01
02
03
04
-
-
-
-
-
-
-
-
-

-
-

-
-
-
II
III
IV
M: Male F: Female +: Presence of activity -: Absence of activity

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TOXICITY OF POLYTRIN-C ON THE PSYCHOPHARMACOLOGICAL
OBSERVATIONS IN WISTAR RAT.

Table 3
Motor activity (counts/15min) in Wistar rats treated with different
concentration of polytrin-D.

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Group Sex
Week
I II III IV
I Control
Male
199.2a
±
3.78
208.1a
±
4.08
202.4a
±
5.03
188.2b
±
3.38
198.1a
±
3.45
164.6c
±
2.88
178.6b
208.0a
±
2.99
199.0a
±
4.88
178.6b
±
5.17
173.2b
±
3.12
187.0b
±
5.53
171.0b
±
4.46
171.1a
±
7.04
163.4a
±
4.68
185.2a
±
3.96
196.4b
±
6.06
180.5a
±
1.92
166.4a
±
2.28
146.2b
186.0a
±
6.82
211.6a
±
4.07
190.0a
±
4.06
184.0b
±
7.12
173.2c
±
3.01
153.2c
±
3.81
132.0d
±
2.94
123.5d
±
3.08
Female
II 8.5 Mg/kg b.w.
Male
Female
III 17.0 Mg/kg b.w.
Male
Female
IV 34.0 Mg/kg b.w.
Male
±
3.11
168.2c
±
2.61
163.0c
±
3.32
152.0c
±
3.15
±
4.78
134.1c
±
2.17
Female
Values are presented as mean ± Standard Error
Values (sex-wise) carrying similar superscripts are not statistically significant (p>0.05).

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TOXICITY OF POLYTRIN-C ON THE PSYCHOPHARMACOLOGICAL
OBSERVATIONS IN WISTAR RAT.

Table 4.
Gripping strength (counts) in Wistar rats treated with different concentration of
Polytrin-D.
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Group Sex
Week
I II III
29.4a
±
1.87
27.0a
±
1.91
32.0a
±
1.32
26.0a
±
0.94
28.2a
±
0.84
26.8a
±
1.11
24.8b
±
0.87
19.0b
±
1.18
IV
32.0a
±
1.02
30.3a
±
1.17
31.2a
±
2.04
28.0a
±
1.09
25.0b
±
1.04
24.0b
±
1.81
23.0b
±
1.07
17.0b
±
2.01
I Control
Male
34.2a
±
1.81
29.0a
±
1.13
28.0b
±
1.72
29.4a
±
0.81
33.0a
±
1.17
31.0a
±
0.91
28.5b
±
1.01
26.2b
±
1.14
31.0a
±
0.94
32.1a
±
1.26
30.0a
±
1.11
30.5a
±
1.13
27.0b
±
1.22
29.0b
±
0.91
27.0b
±
1.24
23.0c
±
1.06
Female
II 8.5 Mg/kg b.w.
Male
Female
III 17.0 Mg/kg b.w.
Male
Female
IV 34.0 Mg/kg b.w.
Male
Female

Values are presented as mean ± Standard Error
Values (sex-wise) carrying similar superscripts are not statistically significant (p>0.05).

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TOXICITY OF POLYTRIN-C ON THE PSYCHOPHARMACOLOGICAL
OBSERVATIONS IN WISTAR RAT.

DISCUSSION
In the present study there was a decrease in
the body weight gain in rats treated with polytrin-C
at the dose of 17 and 34 mg/kg b.w., which was
similar to the observation in two different 2-year
studies carried out with cypermethrin in rats15,16. But
opposed to the findings (mentioned earlier) of these
2-year studies with cypermethrin, the present study
with the combination pesticide polytrin –C did show
clinical signs of intoxication.
Clinical signs of neurotoxicity reported by
earlier studies include impaired ability to walk
abnormal gait, splayed hind limbs, lethargy,
piloerection, ataxia, paralysis, hypersensitivity, gross
disorientation and convulsions17 mild signs of
neurotoxicity like lethargy, hypersensitivity and lack
of grooming were observed in the high dose group
(34 mg polytrin-C/kg b.w.) in the present study but
severe signs of neurotoxicity like ataxia, tremors,
convulsions and paralysis were not observed in the
polytrin-C treated groups.
From the 28- day study polytrin-C, it was
clear that the combination pesticide exhibited
comparatively lesser toxicity than the constitute
pesticides. This phenomenon could be due to the
lower availability of profenofos or cypermethrin
when administered as a combination or could be due
to the relatively lower toxicity of the combination
pesticide polytrin-C compared to the constitute
insecticides profenofos and cypermethrin.

SUMMARY
The combination pesticides are gaining
popularity in pest control programmes as they
exhibit a broad spectrum of activity coupled with
better efficiency and economy. But for the
registration of these pesticides, acute toxicity data is
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sufficient and therefore the long-term toxicity of
these compounds remains unexplored. Alternatives
are required for long-term studies as they are
difficult to carryout, time consuming and expensive.
From this study it can be understood that a well
designed sub-acute
neurotoxicological assessment can provide a major
part of the information observed from the long term
study18. The study also revealed that the
combination pesticide behave differently and exhibit
a different toxicological profile when compared with
the toxicity of the individual pesticide in the
combination. Therefore it is very necessary that all
the toxicological studies have to be carried out (as
for a new pesticide) for a combination pesticide
before it can be brought to the field. Similar studies
can also help to identify combination pesticides that
have lesser mammalian toxicity compared to the
individual pesticides used in the combination while
retaining or obtaining better efficiency.
The present study was carried out to evaluate
the short-term sub acute (28 days) toxicity and
neurotoxicity of Polytrin-C in Wistar rats. The acute
toxicity of the combination pesticide was evaluated
to arrive at the doses for the sub acute study. Sub
acute psychopharmalogical studies (28 days) such as
spontaneous motor activity and muscle relaxant
property were also observed to estimate the
neurotoxicity of these pesticides.
Rats administered polytrin-C [profenofos
40% + cypermethrin 4%] for 28 days at the doses of
8.5, 17 and 34 mg/kg b.w., showed a decrease in
feed consumption and a related decrease in body
weight compared to control, from the third week of
treatment. Psychopharmalogical
revealed only minor behavioral abnormalities in rats
treated with polytrin-C at the doses of 17 and 34
mg/kg b.w., from the third week of the study. A
study clubbed with
investigation

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TOXICITY OF POLYTRIN-C ON THE PSYCHOPHARMACOLOGICAL
OBSERVATIONS IN WISTAR RAT.

decrease in spontaneous motor activity and muscle
relaxant property was observed in the highest dose
group of 34 mg/kg b.w., while only minor
inconsistent (not time related) changes were
observed in other group.

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TOXICITY OF POLYTRIN-C ON THE PSYCHOPHARMACOLOGICAL
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18. Sekar Babu Hari Ram PhD Thesis, Faculty of
Biomedical Sciences,
Universality Chennai (2004).
Dr.M.G.R.Medical