Clinical & Experimental Pathology

Hamid et al., J Clin Exp Pathol 2012, 2:7

http://dx.doi.org/10.4172/2161-0681.1000126

Research Article

Open Access

Carbaryl, A Pesticide Causes Reproductive Toxicity in Albino Rats

Sajad Hamid1*, Seema Sharm2 and Shayama Razdan2
1
2

Lecturer Anatomy, SKIMS Medical College, Jammu & Kashmir, India

Professor Anatomy, Government Medical College, Jammu, Jammu & Kashmir, India

Abstract
The key to mans health lies largely in his environment & often man is responsible for the pollution of his environment
through urbanization, industrialization and other human activities for example commercial agriculture and garden pest
control etc. by using pesticides.
Pesticides, though present in the environment in small quantities as compared to other contaminants such as
industrial wastes and fertilizers, account for public and scientific concern due to their high biological activity. In the recent
years, use of carbamate insecticides has gained importance due to ban of the insecticides belonging to Organochlorine
groups that is D.D.T., Aldrin, Lindane and Endosulfan. These pesticides have a tendency to persist and have potential
to bioaccumulate in the body Kamrin [1].
Concern about the susceptibility of the male reproductive system to drugs or environmental agents has assumed
an increasing extent. The outcome of such exposures have included not only reduced fertility but also embryo/fetal
loss, birth defects, childhood cancer, and other postnatal or functional deficits. Carbaryl is one of the most important
insecticides as it is widely produced and used which has prompted us to initiate this study .
The present study was conducted on 40 male Wistar albino rats as experimental animals. The rats were procured
from the Animals House of the Department of Pharmacology, Government Medical College, Jammu. The rats were
divided in four groups as normal control group I, group II, group III and group IV. All the rats were group housed and
were fed with standard pellet diet and water for two weeks. After two weeks, the rats of group I were left as such and
rats of groups II, III and IV were given 50 mg, 100 mg and 200 mg/kg body weight/day of carbaryl drug in 0.2 ml of
groundnut oil orally, 6 days/week for 60 days, respectively. After 60 days, all the rats were anaesthetized by keeping
them in an inverted glass jar containing large piece of cotton soaked in anaesthetic ether. The testis were dissected
out from each rat and were cut into smaller pieces. These pieces were immediately fixed in 10% formalin. The blocks
were prepared for section cutting with a microtome by paraffin wax embedding method. The sections of 7 thickness
were mounted on glass slides and were stained by H&E and Massons trichome stain.
The following findings are drawn from the study:(1) There is variation in the shape of seminiferous tubules of testis.
(2) There is sloughing of the germinal cells from the basement membrane.
(3) There is depressed spermatogenesis and loss of sperms.
(4) Some tubules are showing accumulation of cellular masses in the lumen of seminiferous tubules of testis.
(5) Interstitial spaces are showing the oedema.
(6) Degenerated Leydig cells are also seen.
These findings are highly conclusive of reproductive toxicity produced by an insecticide, Carbaryl. It is concluded
that the toxic effects are more pronounced in the peripheral parts of the sections of testis. Moreover, intensity of toxic
effects both in peripheral and central parts increases with increase in dosage of the carbaryl drug.

Keywords: Albino rats; Testis; Carbaryl; Depressed spermatogenesis

Introduction
The key to mans health lies largely in his environment. Infact,
much of mans ill health can be traced to adverse environmental
factors, such as water pollution, soil pollution, air pollution etc. which
pose a constant threat to mans health. Often man is responsible for the
pollution of his environment through urbanization, industrialization
and other human activities for example commercial agriculture and
garden pest control etc by using pesticides.
Pesticides, though present in the environment in small quantities
as compared to other contaminants such as industrial wastes and
fertilizers, account for public and scientific concern due to their
high biological activity. According to the Stockholm Convention
on Persistent Organic Pollutants, 9 of the 12 most dangerous and
persistentorganic chemicals are pesticides [2]. It is classified as a likely
J Clin Exp Pathol
ISSN: 2161-0681 JCEP, an open access journal

human carcinogen by the United States Environmental Protection

Agency(EPA.) [3].
Pesticides is the general term for insecticides, acaricides,
rodenticides, herbicides, fungicides, etc. They are widely used in

*Corresponding author: Sajad Hamid, Lecturer Anatomy, SKIMS Medical

College, Bemina, Srinagar, Jammu & Kashmir, India, Tel: 9419506978; E-mail:
drsajadk@rediffmail.com
Received August 08, 2012; Accepted August 31, 2012; Published September
03, 2012
Citation: Hamid S, Sharm S, Razdan S (2012) Carbaryl, A Pesticide Causes
Reproductive Toxicity in Albino Rats. J Clin Exp Pathol 2:126. doi:10.4172/21610681.1000126
Copyright: 2012 Hamid S, et al. This is an open-access article distributed under
the terms of the Creative Commons Attribution License, which permits unrestricted
use, distribution, and reproduction in any medium, provided the original author and
source are credited.

Volume 2 Issue 7 1000126

Citation: Hamid S, Sharm S, Razdan S (2012) Carbaryl, A Pesticide Causes Reproductive Toxicity in Albino Rats. J Clin Exp Pathol 2:126.
doi:10.4172/2161-0681.1000126

Page 2 of 7

industry, agriculture and for public health purposes. Unfortunately,

pesticides are toxic to a greater or lesser extent towards non-target
organisms, including humans Ernest and Patricia [4].

forests, lawns, ornamentals, shade trees and other crops as well as

poultry livestock and pets. It is also used as molluscicide and acaricide
Rani et al. [5].

The synthetic chemical pesticides can structurally be classified into

the following groups:

Carbaryl can produce adverse effects in humans by skin contact,

inhalation or ingestion. Direct contact of skin or eyes with moderate
levels can cause burns. Inhalation or ingestion of very large amounts
can be toxic to nervous and respiratory system.

Organochlorine pesticides
Organophosphate pesticides
Carbamates
Synthetic pyrethroids
Miscellaneous.

Benefits from Pesticides

The important contributions of pesticides to our heath and
economy guarantee their continued use as a class, so require the most
complete knowledge of their toxicology that we can achieve in order to
avoid hazards.
They contribute directly to our health through control of certain
vector-borne diseases and to the economy through increased
production of food. Nearly two-third of world population does not
have sufficient food when more than 30% of our food crops continue
to be destroyed by pests.
Roughly 55,000 tons of pesticides are annually applied in field
crops. Of this 40,000 tons are insecticide, 8000 tons are fungicides and
the rest herbicides. On an average the pesticide used in India is one
tenth of what is applied per unit area of cropped field in the advanced
countries.

Pesticides as Environmental Pollutants

Pesticides hold a unique position among environmental
contaminants, being present in the environment in such small
quantities as compared to other contaminants such as industrial wastes
and fertilizers. The major factors which account for public and scientific
concern is their biological activity.

Role of Pesticides on Testis

Of the potential health risks associated with exposure to chemical
or physical agents, a prominent concern is that these agents may
interfere with the ability of individuals to produce normal, healthy
children. A large number of chemicals that have been released into
the environment are known to interfere with the endocrine system.
Sexual development during the prenatal and neonatal period is under
hormonal control and is, therefore, sensitive to exogenous substances
with an endocrine effect. Keeping in mind the pivotal role of testis in
reproduction this experimental work was done.
Attention in this thesis primarily is focused on toxic effects that
involve testicular and postspermatogenic processes that are essential
for reproductive success.

Choice of Pesticides for the Present Study

Carbaryl was taken to see the effect on testis of albino rats. Carbaryl
which is 1-naphthyl-N-methyl carbamate is a broad-spectrum
insecticide used to protect vegetables, cotton, fruits, cereals and other
crops against a variety of insects and pests. Carbaryl is 1-naphthyl-Nmethyl carbamate and was first synthesized in 1953 and introduced
in 1958. It controls over 100 species of insects on citrus fruits, cotton,
J Clin Exp Pathol
ISSN: 2161-0681 JCEP, an open access journal

Pesticides clearly have the potential to cause reproductive toxicity

in animals and several compounds are known to affect human
reproduction [6,7].
Epidemiological studies postulated that in the past 50 years the
sperm number and sperm quality in human had been decreased [8,9].
Pathological effects of pesticides on the reproductive system of
experimental animals were recorded by many authors [10-13].
Different studies have been done to see the effects of carbamate
exposure on the testis of experimental animals.
Degraeve et al. [14] found that in mice injected intraperitoneally
with 0.4 mg of carbaryl daily for one week, the incidence of sperm
abnormalities were reportedly increased, but no degenerative changes
in the testis were seen.
Kitagawa et al. [15] reported reduced numbers of spermatogonia
and spermatozoa in rats given 3 mg of carbaryl per week orally for one
year.
Rani et al. [5] found marked histopathological changes in testis of
albino rats following oral administration of carbaryl in dose of 100 and
200 mg/kg body weight in 0.2 ml of groundnut oil orally, 6 days/week
for 60 days.
The light microscopic observation demonstrated distorted shape
of seminiferous tubules, disturbed spermatogenesis, accumulation of
cellular mass in the lumen of tubules, oedema of interstitial spaces and
loss of sperms of varying degrees and detachment of germ cells from
the basement membrane. Same findings were also reported in [16-19].
Nonetheless, carbamate use may pose a significant risk of poisoning
if handled carelessly. Health professionals may need to assess the
consequences of prior exposure and should understand the fate of
these compounds after absorption by humans.
The deleterious effects of carbaryl on reproductive system have
prompted us to perform this study. The present study is aimed to
evaluate the effect of carbaryl on the histomorphological characteristics
of the testis in a mammal, the albino rat.
Vashakidze [20] reported teratogenecity and decreased
reproduction at subchronic intubated dosages of 100 mg kg/day and
higher but not at 50 mg/kg/day. However, a single 50 mg/kg/day in
intubated dose on gestation day nine or ten was reported to cause
teratogenecity.
Robens [21] observed that carbaryl administered to guinea pigs
by gavage at 300 mg/kg/day (a toxic dosage) during organogenesis
resulted in fetal skeletal anomalies; fetal mortality resulted from dosing
on gestation days 11-21, but not from single doses during gestation.
Dikshith et al. [22] demonstrated that oral administration of
carbaryl (200 mg/kg for 3 days a week) for a period of 90 days did
not produce any overt toxicological signs in male rats. There were no
significant histological changes in testis, epididymis, liver and kidney.
Similarly, no marked biochemical changes were seen in testis, liver and

Volume 2 Issue 7 1000126

Citation: Hamid S, Sharm S, Razdan S (2012) Carbaryl, A Pesticide Causes Reproductive Toxicity in Albino Rats. J Clin Exp Pathol 2:126.
doi:10.4172/2161-0681.1000126

Page 3 of 7

brain. The activity of acetycholine esterase in blood of carbaryl treated

rats was however, found to be decreased. Carbaryl did not affect the
fertility of male rats at 200 mg/kg up to 90 days.
Murray et al. [23] showed that when carbaryl administered to mice
by gavage at 100 mg/kg/day on gestation days 6-15 had no fetotoxic
effects, dietary exposure at 5660 ppm (1166 mg/kg/day) resulted in
decreased fetal size but no teratogenecity.
Martin [24] showed that when carbaryl administered to mice for 5
days upto 800 mg/kg/day or by gavage at 150 mg/kg/every 2 days for
upto 68 days, did not affect testis weight, histology, sperm count and
frequency of sperm abnormalities.
Osterloh et al. [25] found that when carbaryl administered to
mice up for 5 days at upto 800 mg/kg/day did not affect testis weight,
histology sperm count and frequency of sperm abnormalities.
Sever and Hessol [26] stated that Carbaryl given to mice at upto 34
mg/kg/day for 5 days did not affect either the weight of the testis and
sex glands or the ability of the prostate to assimilate and metabolize
testosterone.

Material and Methodology

The present study is based on the findings carried out on 40 male
Wistar albino rats as experimental animals.

Collection of material
Healthy male Wistar albino rats weighing between 50-80 grams
were obtained on December 13, 2010 from the Animals House of the
Department of Pharmacology, Medical College Jammu. Forty rats were
included in this study.

Grouping of animals

such. Whereas, the drugs were administered to other groups as shown

in (Table 1).

Dissection of experimental animals

After 60 days all the rats were sacrificed after anaesthetizing them
in an inverted glass jar containing large piece of cotton soaked in
anaesthetic ether and dissection of testis of albino rats was done on the
same day.
The scrotum was skinned and a midline incision was given with
the help of scalpel and forceps. The testis were dissected out from the
scrotum of each rat. The naked eye examination was done to see any
external changes. The dissected out testis were cut into smaller pieces
(5 mm) and were kept in tissue capsules along with a label indicating
the serial groups I, II, III and IV.

Methodology
Manual processing of tissues
The casting and embedding was done with the help of moulds. Two
L-shaped blocks were placed on a metallic plate, which acts as a base of
the mould and molten wax was poured into it. The tissues were placed
in the mould filled with wax and left to solidify. After solidification the
blocks of the wax were removed and properly labeled for microtomy.
The slides were subsequently stained by a haematoxylin and eosin &
Massons trichrome. The slides were cleaned beyond the area of tissue
implantation, dried and mounted in DPX and examined first under
low power and then high power (Table 2).

Results
Observations in normal control group I (Figure 1)

The rats were divided into the following four groups and
identification number was given to the rats of each group.
Group I: Normal control - 10 rats

Macroscopic changes: No gross changes seen.

Microscopic changes: Cut sections of testis in various planes show
that the testis contain two components as:

Group II: Experimental group - 10 rats

(A) Seminiferous tubules with many cells thickened walls.

Group III: Experimental group - 10 rats

(B) Interstitial cells disposed in small groups in connective tissue

stroma between the tubules.

Group IV: Experimental group - 10 rats

All the rats were group housed in small iron cages in a room,
where temperature was maintained at 23 1C. The rats were fed with
standard pellet diet and water for two weeks.
After two weeks the rats of normal control group I were left as
Experimental groups

Observations in carbaryl (50 mg/kg/day) treated rats group

II (Figure 2)
The rats fed with 50 mg/kg/day for 60 days show following features
in the testis:-

Date of administration of 1st dose of Route of administration

carbaryl drug
of carbaryl drug

Duration of carbaryl drug

given

Dose of carbaryl drug administered to each

group

Group II

28/12/2010

Oral

6 days/ week for 60 days

50 mg/kg body weight/ day in 0.2 ml of

groundnut oil

Group III

28/12/2010

Oral

6 days/ week for 60 days

100 mg/kg body weight/ day in 0.2 ml of

groundnut oil

Group IV

28/12/2010

Oral

6 days/ week for 60 days

200 mg/kg body weight/ day in 0.2 ml of

groundnut oil

Table 1: Administration of Carbaryl Drug To Experimental Groups.

S. No

Step

Fixation

Medium
10% formal saline

Dehydration

Acetone

3 changes at intervals of 2 hours

3 changes at intervals of 2 hours

Clearing

Benzene

Wax embedding

Paraffin wax at 56OC

Type
12 hours

Two changes in molten wax at its melting point were given at time intervals of 1 hrs. & 1 hr respectively
Table 2: Manual processing of tissues.

J Clin Exp Pathol

ISSN: 2161-0681 JCEP, an open access journal

Volume 2 Issue 7 1000126

Citation: Hamid S, Sharm S, Razdan S (2012) Carbaryl, A Pesticide Causes Reproductive Toxicity in Albino Rats. J Clin Exp Pathol 2:126.
doi:10.4172/2161-0681.1000126

Page 4 of 7

Macroscopic changes: No gross changes seen.

Microscopic changes: (A) Changes in seminiferous tubules

D

B
F

C
ii

A
E

Mild to moderate degenerative changes in spermatogenic cells.

In addition focal individual cell necrosis is also seen especially

towards the lumen of tubules.

iii

Figure 1: (i) Testis of group I albino rat (control group) showing seminiferous
tubules (A), Tunica albuginea (B), basement membrane(C) (h & e stain x
100), (ii) Ts of seminiferous tubule of testies of group I albino rat (control
group) showing basement membrane (A) Spermatogonia (B) Primary
spermatocyte (C) Secondary spermatocyte (D), Spermatids (E) Tails of
spermatids (F) Sertoli cell (G) (h & e stain x 400), (iii)Testis of group I albino
rat (control group) showing basement membrane of seminiferous tubule (A)
Spermatogonia (B) Primary spermatocyte (C) Secondary spermatocyte (D)
Spermatids (E) Tails of spermatids (F) Blood vessel in interstitial space (G)
(massons trichome x 400).

The other changes seen are: collection of cellular masses in

their lumen (few sections).
Detachment of the basement membrance from the seminiferous
epithelium (in few of the seminiferous tubules).

G
C D

Seminiferous tubules show mild variation in their size with

focal distortion of normal shape which is mostly observed in
peripheral seminiferous tubules (only some of the seminiferous
tubules).

D C

ii
B

Interstitial oedema is observed in the interstitial spaces which

is more prominent in the peripheral region of testis.

Macroscopic changes: No gross changes seen.

Seminiferous tubules show distortion in the normal size.
Variation in size is more marked than the previous group
and is mostly observed in peripheral tubules, (in many of the
seminiferous tubules).

Microscopic changes: Changes in seminiferous tubules

iii

Interstitial cells of Leydig show mild to moderate degenerative

changes in the interstitial spaces in between the seminiferous
tubules.

The group of rats fed with 100 mg/kg/day for 60 days show
following observations:-

(B) Changes in the interstitium

Observations in carbaryl (100 mg/kg/day) treated rats group

III (Figure 3)

In few of tubules, spermatids and spermatozoa are identified.

A
iv

Collection of cellular masses in the lumen of seminiferous

tubules (several sections).
Separation of the basement membrane from the spermatogenic
series of cells, which is more marked than the findings seen
in the rats given with low doses of carbaryl ( in several of
seminiferous tubules).

C
B

Figure 2: (i)Ts of testis of group II albino rat after giving carbaryl 50 mg/
kg body wt orally showing mild variation in shape of seminiferous tubules
(A) Collection of cellular masses in lumen of seminiferous tubules (B) Mild
interstitial oedema (C) Degenerated interstitial cells (D) (h & e stain x 100),
(ii) Ts of testis of group II albino rat after giving carbaryl 50 mg/kg body wt
orally showing detached basement membrane from germ cells (A) Focal cell
necrosis towards lumen of seminiferous tubules (B), Mild interstitial oedema
(C) (h & e stain x 100), (iii)Ts of testis of group II albino rat after giving carbaryl
50 mg/kg body wt orally showing mild degenerative changes in interstitial
cells of leydig (A), interstitial oedema (B), cell necrosis (C) (h &e stain x100),
(iv)Ts of testis of group II albino rat after giving carbaryl 50 mg/kg body wt
orally showing broken basement membrane (A) Interstitial oedema (B) (h & e
stain x 100), (v) Ts of testis of group II albino rat after giving carbaryl 50 mg/kg
body wt orally showing mild variation in shape in few of seminiferous tubules
(A) Collection of cellular masses towards lumen of seminiferous tubules (B)
Focal cell necrosis towards lumen of seminiferous tubules (C) (massons
trichome x 100).

J Clin Exp Pathol

ISSN: 2161-0681 JCEP, an open access journal

Moderate to severe degenerative changes in spermatogenic

cells (in several sections of seminiferous tubules).
Marked cell necrosis in seminiferous tubules towards the
lumen.
Many of tubular sections show loss of spermatids and
spermatozoa.
(B) Changes in interstitium
Moderate degenerative changes in the interstitial cells of
Leydig.
Marked interstitial oedema especially in the peripheral region of
the testis. The interstitial oedema is more marked as compared
to the findings seen in rats given with (50 mg/kg/day).

Volume 2 Issue 7 1000126

Citation: Hamid S, Sharm S, Razdan S (2012) Carbaryl, A Pesticide Causes Reproductive Toxicity in Albino Rats. J Clin Exp Pathol 2:126.
doi:10.4172/2161-0681.1000126

Page 5 of 7

Dilated blood capillaries are observed in the focal areas in the

interstitium of the seminiferous tubules.

Observations in carbaryl (200 mg/kg/day) treated rats group

IV (Figure 4)

The following findings are seen in the testis of rats fed with 200 mg/
kg/day for 60 days:-

ii

Macroscopic changes: No gross changes seen.

Microscopic changes: (A) Changes in the seminiferous tubules
Seminiferous tubules show small atrophied seminiferous
tubules with marked distortion in their shape and size(multiple
sections). Variations seen in their shape are more pronounced
than the previous groups with low doses of carbaryl.
The seminiferous tubules show cell necrosis towards the lumen
of tubules which is more marked than the previous groups.
These changes are more prominent in the peripheral tubules
of testis.
Sloughing of the germinal cells from the basement membrane
(in several seminiferous tubules).
The spermatogenesis is much disturbed as compared with the
previous groups with low doses of carbaryl.
Loss of spermatids and spermatozoa (in many seminiferous
tubules).

A
A

iv
iii

B
A

Figure 4: (i)Ts of testis of group IV albino rat after giving carbaryl 200 mg/
kg body wt orally showing (A)Atrophied seminiferous tubules with marked
interstitial oedema (B) and cell necrosis (C) (h & e stain x 100), (ii) Ts of
testis of group IV albino rat after giving carbaryl 200 mg/kg body wt orally
showing marked variation in shapes of seminiferous tubules (A) Detachment
of basement membrane from germ cells (B) (h & e stain x 100, (iii)Ts of testis
of group IV albino rat after giving carbaryl 200 mg/kg body wt orally showing
interstitial oedema more marked towards periphery (A) Cell necrosis (B)
(h & e stain x 100), (iv)Ts of testis of group IV albino rat after giving carbaryl
200 mg/kg body wt orally showing atrophied seminiferous tubules (A) Marked
variation in the shape of seminiferous tubules (B) Marked Interstitial oedema
(C) (massons trichome x 100, (v)Ts of testis of group IV albino rat aftergiving
Carbaryl 200 mg/kg body wt orally showing distortion of seminiferous tubule
(A) Disturbed spermatogenesis (B) (massons trichome x 100),

Accumulation of cellular masses in their lumen (in several

seminiferous tubules).

ii

Detachment of germ cells from the basement membrane.

This detachment of basement membrane is more marked as
compared with groups with low doses of carbaryl.

B
C

C
B

iii

iv

A
B

(B) Changes in the interstitium:

Oedema of the interstitial spaces (more marked in the
peripheral region). Interstitial oedema is also much significant
in this group compared with other groups.
Prominent degenerative changes in the Leydig cells compared
to other groups with low doses of carbaryl.

Figure 3: (i) Ts of testis of group III albino rat after giving carbaryl 100 mg/kg
body wt orally showing marked variation in shape of seminiferous tubules of
testis (A) Dilated blood capillaries in the interstitiun of testis (B) (h & e stain x
100), (ii) Ts of testis of group III albino rat after giving carbaryl 100 mg/kg body
wt orally showing collection of cellular masses in lumen of several sections of
seminiferous tubules (A) Loss of germinal layers of cells of seminifous tubule
more marked towards periphery (B) (h & e stain x 100), (iii) Testis of group
III albino rat showing severe degenerative changes in spernatogenic cells
(A) Loss of spermatids and spermatozoa (B) Interstitial oedema (C) (h & e
stain x 100), (iv) Testis of group III albino rat after giving carbaryl 100 mg/
kg body wt orally showing marked interstitial oedema (A) Cell necrosis (B) (h
& e stain x 100), (v) Testis of group III A rat after giving carbaryl 100 mg/kg
body wt orally showing interstitial oedema (A) Interstitial cells degeneration
(B) (massons trichome x 100).

J Clin Exp Pathol

ISSN: 2161-0681 JCEP, an open access journal

Discussion
An extremely complex mechanism underlies the effects of
various substances on reproductive components and functions.
Various chemicals may interfere in different ways with components
of reproductive system. They may affect directly by interference of
the substance with reproductive components or indirectly by altering
hormonal regulations.
The carbamate insecticides, one of which is Carbaryl, exert their
insecticidal action by inhibiting cholinesterase enzymes. This inhibition
is the primary mechanism by which these insecticides cause toxicity
in mammals. The cholinesterase enzymes hydrolyze acetycholine

Volume 2 Issue 7 1000126

Citation: Hamid S, Sharm S, Razdan S (2012) Carbaryl, A Pesticide Causes Reproductive Toxicity in Albino Rats. J Clin Exp Pathol 2:126.
doi:10.4172/2161-0681.1000126

Page 6 of 7

and other choline esters; consequently, their inhibition leads to the

accumulation of endogenous acetycholine and other choline esters.
Probably most of the biologic effects of anticholinesterase agents,
including carbaryl, are due to the inhibition of acetylcholinesterase
which leads to the accumulation of endogenous acetycholine, the
principal choline ester that has demonstrated physiologic significance
in humans.
The present study shows distorted shape of seminiferous tubules,
disturbed spermatogenesis, accumulation of cellular mass in the lumen
of seminiferous tubules, oedema of interstitial spaces, loss of sperms
of varying degrees and detachment of germ cells from the basement
membrane of seminiferous tubles of testis. Same findings were
also reported by Rani et al. [5]. In the testis of albino rats following
administration of carbaryl in dose of 100 and 200 mg/kg body weight in
0.2 ml of groundnut oil orally/6 days week for 60 days. Similar findings
were also reported in [16-19], who have reported spermatotoxic effect
of carbaryl in adult and young male rats given with 50 and 100 mg/kg
body weight. Male fed 5 days/week for 60 days, caused dose and agedependent decline in epidydmal sperm count and sperm motility and
an increase in number of sperms with abnormal morphology. Young
animals in comparison to adults exhibited pronounced spermatotoxic
effects. Some of these findings are in accordance with the present
study in which the dose-related decreased spermatogenesis and loss of
sperms of varying degrees have been found.
The present study reveals marked histomorphological and
degenerative changes of the lining cells of seminiferous tubules.
These findings are in accordance with [16], who observed deleterious
effects of chronic and subchronic administration of carbaryl on
male reproductive system of animals. These effects included damage
to the germinal epithelium of seminiferous tubules and altered
spermatogenesis.
Vashakidze [20] reported teratogenecity and decreased
reproduction at subchronic intubated dosages of 100 mg/kg body
weight/day and higher but not at 50 mg/kg/day. However, a single 50
mg/kg/day in intubated dose on gestation day nine or ten reported to
cause teratogenecity. These findings correlate with the present study in
which dose related effect is seen on spermatogenesis and loss of sperms
of varying degrees in testis of male albino rats.
Kitagawa et al. [15] reported reduced number of spermatogonia
and spermatozoa in rats given 3 mg/kg body weight of carbaryl orally
for 1 year. These findings are also in accordance with the present study
showing depressed spermatogenesis in rats fed with carbaryl orally.
The present study done on male albino rats by giving them
carbaryl in dose of 50, 100 and 200 mg/kg body weight in 0.2 ml of
groundnut oil orally/6 days week for 60 days showed distorted shape
of seminiferous tubules and significant histomorphological changes
in testis of albino rats. These findings are contrary to the findings by
Dikshith et al. [22], who demonstrated that oral administration of
carbaryl (200 mg/kg body weight for 3 days a week) for a period of 90
days did not produce any overt toxicological signs in male albino rats.
There were no significant histological changes in testis, liver and brain.
The activity of acetylcholine esterase in blood of carbaryl treated rats
was however found to be decreased. Carbaryl did not affect the fertility
of male albino rats at 200 mg/kg body weight upto 90 days.
Martin [24], Osterloh et al. [25] showed that when carbaryl
administered to mice up for 5 days at upto 800 mg/kg/day or by gavage
at 150 mg/kg/every 2 days for upto 68 days, did not affect testis weight,
histology, sperm count and frequency of sperm abnormalities. These
J Clin Exp Pathol
ISSN: 2161-0681 JCEP, an open access journal

findings are not in accordance with the present study which reveals the
distorted shape of seminiferous tubules, disturbed spermatogenesis,
loss of sperms in the testis of male albino rats. These findings are much
significant with the high dose of carbaryl.
Carpenter et al. [27] reported that carbaryl in the diet of rats for
2 years at 400 ppm (about 20 mg/kg/day) slightly depressed organ
weights in males but did not affect mortality, haematology or organ
histopathology. No effect levels were at 9 mg/kg/day for males and 21
mg/kg/day for females. In shorter term studies at higher dosages, liver
and kidney effects were noted, which were transient and may have been
secondary to stress. Though our study does not affect mortality yet it
shows significant histopathological changes on testis of albino rats.
Benson and Dorough [28] observed that when rats and gerbils
were given carbaryl orally for 70 days at dosages that were increased
weekly, all deaths occurred within 24 hours of the first administration
of a given dosage. One of 12 rats died at dosage of 120 mg/kg/day, and
cumulative mortality was 7/12 at a dosage of 180 mg/kg/day; no further
deaths occurred at dosages of upto 200 mg/kg/day. In gerbils, mortality
was 2/12 at the initial dosage of 60 mg/kg/day, and the last animal died
at a dosage of 100 mg/kg/day. On the contrary there is no mortality of
albino rats in the present study at doses of 50, 100 and 200 mg/kg body
weight respectively.
The present study shows altered spermatogenesis which can lead to
infertility by giving carbaryl to male albino rats. These findings do not
correlate with the findings seen by Orlova and Zhalbe [29]. They could
not find any change in fertility, gestation and viability of carbaryl (2
mg/kg) treated pups and rats. Similarly, studies of Benson and Dorough
[28] showed that carbaryl (10 and 30 mg/kg body weight) induced no
reproductive or teratogenic effects in mice. Weil et al. [30] observed no
significant effects of carbaryl (10 mg/kg) on fertility, gestation, viability
or lactation of rats. Studying non-human primates, Dougherty et al.
[31] also could not find any teratogenic effects of carbaryl (2 and 20
mg/kg) in rhesus monkeys.
In the present study there is decreased sperm count. This is
supported by decreased sperm number, postulated in the past 50 years
in epidemiological studies [8,9].
Considering the effects of carbaryl on the testis in the present
study and based on the findings of earlier studies, this compound may
be designated as moderately toxic. This may affect spermatogenesis
resulting in the production of decrease number of sperms.

Conclusion
Carbaryl is being used extensively as a broad spectrum pesticide.
It is known toxicant to the male reproductive system and, is therefore,
under focus in the present study. The results of the present study have
thrown some light on the toxic effects of carbaryl on testicular functions
that are essential for reproductive success.
The present study was conducted on 40 male Wistar albino rats as
experimental animals. The rats were procured from the Animals House
of the Department of Pharmacology, Government Medical College,
Jammu. The rats were divided in four groups as normal control group
I, group II, group III and group IV. All the rats were group housed
and were fed with standard pellet diet and water for two weeks. After
two weeks, the rats of group I were left as such and rats of groups II,
III and IV were given 50 mg, 100 mg and 200 mg/kg body weight/
day of carbaryl drug in 0.2 ml of groundnut oil orally, 6 days/week for
60 days, respectively. After 60 days, all the rats were anaesthetized by
keeping them in an inverted glass jar containing large piece of cotton

Volume 2 Issue 7 1000126

Citation: Hamid S, Sharm S, Razdan S (2012) Carbaryl, A Pesticide Causes Reproductive Toxicity in Albino Rats. J Clin Exp Pathol 2:126.
doi:10.4172/2161-0681.1000126

Page 7 of 7

soaked in anaesthetic ether. The testis were dissected out from each
rat and were cut into smaller pieces. These pieces were immediately
fixed in 10% formalin. The blocks were prepared for section cutting
with a microtome by paraffin wax embedding method. The sections of
7 thickness were mounted on glass slides and were stained by H&E
and Massons trichome stain.
The following findings are drawn from the study:(1) There is variation in the shape of seminiferous tubules of testis.
(2) There is sloughing of the germinal cells from the basement
membrane.
(3) There is depressed spermatogenesis and loss of sperms.
(4) Some tubules are showing accumulation of cellular masses in
the lumen of seminiferous tubules of testis.
(5) Interstitial spaces are showing the oedema.
(6) Degenerated Leydig cells are also seen.
However, since the study was conducted on experimental animals
and results may not be exactly the same in humans, suffering from the
carbaryl toxicity. But in no case it can be overlooked, while designing
a therapy for pesticides, where it becomes necessary to take into
consideration the effects of carbaryl on these tissues of vital importance.
It is concluded that the toxic effects are more pronounced in the
peripheral parts of the sections of testis. Moreover, intensity of toxic
effects both in peripheral and central parts increases with increase in
dosage of the carbaryl drug.
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