International Journal of Scientific Research in Environmental Sciences, 2(9), pp.

308-322, 2014
Available online at http://www.ijsrpub.com/ijsres
ISSN: 2322-4983; 2014 IJSRPUB
http://dx.doi.org/10.12983/ijsres-2014-p0308-0322


308
Full Length Research Paper

Carbofuran-Induced Alterations in Body Morphometrics and Histopathology of
Liver and Kidneys in the Swiss Albino Mice Mus Musculus L.
M. Saiful Islam
1
*, Moni Krishno Mohanta
1
, Ananda Kumar Saha
1
, Anup Mondol
1
, M. Mizanul Hoque
1
, Apurba
Kumar Roy
2


1
Genetics and Molecular Biology Laboratory, Department of Zoology, University of Rajshahi, Rajshahi 6205, Bangladesh
2
Department of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi 6205, Bangladesh
*Corresponding author; email: saifulzoo.ru@gmail.com

Received 03 June 2014; Accepted 08 August 2014

Abstract. Carbofuran-fed changes in body, liver and kidney weights as well as alterations in the histopathology of hepatic and
renal tissues in the Swiss albino mice Mus musculaus L. were assessed. Twenty 7-10 days old male mice were divided equally
into five groups viz. T0 (control), T1, T2, T3 and T4, which were allowed to feed on Carbofuran supplemented diets at doses of
0.0 (control), 0.5, 1.0, 1.5 and 2.0 mg/kg food. Compared to the decrease in body and kidney weights of the treated mice,
Carbofuran-fed diets resulted in a significant gain in both body (P<0.004) and kidney weights (P<0.025) but decrease in liver
weight (P<0.001) of the untreated mice. Carbofuran treatments, however, decreased both hepatosomatic and renosomatic
indices of the experimental mice. Histological microphotographs of the liver and kidney tissues of the control showed no
abnormalities in structure, colour, and appearance. While those of the treated mice showed remarkable changes that include
haemosiderin in hepatocytes, epithelial and Kupffer cells, dilated sinusoids, haemorrhage and vacuolation, necrotic hepatocytes
and congested central and portal veins, inflammatory cell infiltration and increased number of mononuclear cells in liver
sections. In kidney sections, on the other hand, oedema, congested or degenerated glomeruli, widened urinary spaces, dilated
or vacuolated tubules, degenerated or eroded walls of Bowmans capsules, haemorrhages and inflammatory cell infiltrations,
hyalinized areas and increased numbers of podocytes and mesangial cells were notable characteristics. Relevance of these
findings to the indiscriminate uses of this carbamate insecticide in agriculture and public health has been discussed.

Keywords: Swiss albino mice, Carbofuran, Morphometrics, Histopathology, Liver, Kidney

1. INTRODUCTION

Carbofuran is a broad spectrum, non-cumulative
carbamate insecticide which is marketed under the
trade names, among others, Furadan
R
, Curaterr
R
, Bay
70143, FMC 10242, ENT-27,164 and Niagara 10242
R

(WHO, 1985). Signs of Carbofuran toxicity in rodents
are generally observed when acetylcholinesterase
activity is inhibited by more than 35% and tremors
occur at inhibition sites by more than 70% (Renzi and
Kreiger, 1986). Carbofuran is a cholinesterase
inhibitor with predominantly contact and stomach
actions and is highly toxic to mammals and other
lower vertebrates including fishes but with no
phytotoxic action (WHO, 1985; Baligar and Kaliwal,
2002). The insecticide is extensively used in
Bangladesh to control insects in a wide variety of field
crops including rice and vegetables (Chowdhury et al.,
2012).
Increasing concern therefore is mounting as
regards the indiscriminate uses of insecticides like
Carbofuran because most of the farmers of
Bangladesh apply the insecticide at 20 kg/ha against
the recommended dose of 16.8 kg/ha (BARC, 1994;
Rahman and Alam, 1997) which obviously impose
risk to non-target biota including fishes, amphibians,
human beings and the environment (Chowdhury et al.,
2012; Uddin et al., 2013). Several reports on
Carbofuran toxicity in mice and rats reveal that the
insecticide induces remarkable histopathological
changes in various organs including liver, stomach,
intestine, spleen, pancreas and kidneys (Rai et al.,
2009; Al-Amoudi, 2012; El-Damaty et al., 2012).
Moreover, effects of another carbamate insecticide
Carbosulfan poisoning in albino mice have been
studied by Ksheerasagar and Kaliwal (2006, 2010).
Since liver is associated with metabolism and
elimination of toxicants from the body and kidney is
associated with excretion, their histological
parameters are considered to be the key points to
elucidate toxicity of the insecticides on the
experimental rodents (Dessouki et al., 2013; El-
Islam et al.
Carbofuran-Induced Alterations in Body Morphometrics and Histopathology of Liver and Kidneys in the Swiss Albino
Mice Mus Musculus L.
309
bendary et al., 2014). This led to design the present
investigation.
Carbofuran-induced damages in a number of
organs and/or systems in rodents have previously been
reported. Thus reproductive organs (Aziz et al., 2008)
and thyroid glands (Hadie et al., 2012) in male rats,
tissues of tail muscles and liver in toad tadpoles
(Jayatillake et al. (2011) and testes (Al-Amoudi,
2012) and testosterone serum concentration in mice
(Elayan et al. (2013) showed remarkable Carbofuran
toxicity.
Apart from the aforesaid carbamate injury and
poisoning in mice and rats, however, a number of
other insecticides and cytotoxic chemicals, for
examples, organochlorine such as Endosulfan
(Aleksandrowiez, 1997; Choudhury et al., 2003),
organophosphate like Chlorpyriphos and Profenofos
(Saha et al., 2006; Dessouki et al., 2013; El-bendary et
al., 2014), pyrethroid like Cypermethrin
(Khalequzzaman et al., 2004; Yavasoglu et al., 2006),
and cisplatin (Adejuwon et al., 2014) have also been
tested to assess their histopathological impacts on the
experimental rodents. Taking these findings in
consideration, the present investigation was designed
to observe a detailed account of Carbofuran-induced
changes in body, liver and kidney weights and
hepatosomatic and renosomatic indices, along with
histopathological alterations in the tissues of liver and
kidneys in the male Swiss albino mice under
laboratory conditions.

2. MATERIALS AND METHODS

2.1. Test animals

Seven to ten days old male Swiss albino mice Mus
musculus L. (Rodentia: Muridae) were collected from
the Animal Resources Branch (ARB), International
Centre for Diarrhoeal Disease Research, Bangladesh
(ICDDR, B). They weighed between 16 and 24g
(mean SD = 21.26 2.34g) and were reared in
groups of four in steel cages of 47cm 37cm 23cm
size with saw dust bedding at 25 2 C and 705%
relative humidity (RH) with 14 hr: 10 hr light: dark
(L:D) regime and were allowed to acclimatize for a
period of 6-7 days prior to experimental use. The
bedding was replaced once a week. The mice were
maintained on a standard diet composed of maize
grain (36.92%), rice polish (18.46%), wheat polish
(24.62%), soybean (18.46%), crude protein (1.23%),
and salt and large grain premix (0.15% each), supplied
twice daily and all the mice had access to drinking
water ad libitum. In compliance with the standard
animal ethical guidelines, the present investigation
was carried out at the Laboratory of Genetics and
Molecular Biology, Department of Zoology,
University of Rajshahi, Bangladesh, during the period
from February 2013 to June 2013.

2.2. Test chemical

Technical grade (99.99%) Carbofuran (product name
Alphafuran 5G) was procured from the Alpha Agro
Limited, Dhaka, Bangladesh.

2.3. Experimental design

Twenty male mice were randomly segregated into five
groups (T0-T4), each having four animals. T0 group
served as the control which received diets without
Carbofuran, whereas groups T1-T4 were given at
doses of 0.5 mg/kg, 1.0 mg/kg, 1.5 mg/kg and 2.0
mg/kg Carbofuran supplemented diet, respectively.
All the experiments were performed in accordance
with the guidelines for use and care of laboratory
animals.

2.4. Body, liver and kidney weights

Initial and final body, liver and kidney weights of the
control and treatment groups of mice were recorded
during the experiment. After 15, 25, 35 and 45 days
post-treatment just before sacrificing the animals, the
morphometric measurements were recoded again. The
mice to be sacrificed were placed in a jar containing
cotton wool soaked in diethyl ether. Complete
anaesthesia was considered accomplished when the
pedal movements and eye lid reflex disappeared and
the animal became recumbent while still breathing.
The belly of the mice was cut open to collect liver and
kidneys which were weighed, gently rinsed in normal
saline, fixed in 10% formalin and stored at 4 C for
histopathology. Finally, hepato- and renosomatic
indices were estimated as follows: liver or kidney
weight body weight 100 (El-Damaty et al., 2012;
Adejuwon et al., 2014).

2.5. Histopathology

The tissue samples of liver and kidney were quickly
excised following the protocol of Sarker et al. (2005).
Slices of the organs were quickly prepared for
histological examination to show if there were
morphological changes in the organs during the
treatment. Processing started with the parking of the
tissues in the tissue capsule. The tissues were
dehydrated in graded levels of ethanol (70-100%) in
ascending order. The alcohol was changed after
soaking the tissues in them for 1 and 2h. The tissue
were cleared in chloroform and impregnated with
paraffin wax and sectioned at 4-5 thickness using a
rotary microtome machine. The sections were floated
International Journal of Scientific Research in Environmental Sciences, 2(9), pp. 308-322, 2014
310
on a water bath maintained at 2-3
o
C below melting
point of paraffin wax. They were on a hot plate
thermostatically maintained at temperature of 2-3
o
C
above the mid point of paraffin wax when properly
dried (15-30 min), they were stained with
haematoxylin and eosin, dehydrated, cleared and
mounted in DPX, avoiding air bubbles. Finally,
examinations of the slides were made by a light
microscope (Zeiss, Germany) and microphotographs
were taken with an automated digital camera system
(Olympus CH30/CH40, Japan) at suitable but varying
magnifications of 4 to 40.

2.6. Statistical analyses

Morphometric data on body, liver and kidney weights
were analyzed using SPSS for Windows (version
19.0) and presented in mean standard deviation.
Experimental data between the control and treatment
groups were subjected to one-way analysis of variance
(ANOVA) followed by the post hoc LSD (least
significant difference) tests to separate the means,
where the levels of significance were set at P<0.05
(Steel and Torrie, 1984).

3. RESULTS

3.1. Morphometric parameters

Body weight: The body weight of mice in the control
group (T0) increased gradually from around 22g to
28g over a period of 45 days, but the body weight
decreased among the treated groups (T1-T4) across
the period of the exposure. Thus Carbofuran induced a
gain in body weight of the untreated mice and the
corresponding loss in the parameter of the treated
mice significantly (F =2.668; P<0.004; Table 1).
Liver weight: There was a slight decrease in liver
weight of the untreated mice (T0) that ranged from
1.85g to 1.60g. Similar trend was also recorded in the
treated mice in a dose-dependent manner; the decline
was from 1.20g to 0.80g over a period of 45 days.
This decline was also reflected in the hepatosomatic
indices from approximately 8% in the control to about
5% in the highest dose. Although the decrease in liver
weight in the experimental mice was statistically
significant (F =3.759; P<0.001; Table 1), however, the
overall decline in the hepatosomatic indices was not
significant (P>0.05).
Kidney weight: Unlike liver weight, the kidney
weight of the control mice increased from 0.55g to
0.62g during the course of the experiment. Whereas
kidney weight of the Carbofuran-treated mice was
found to decrease from 0.52g after 15 days post-
treatment to 0.48g after the termination of the
experiment. Thus the overall increase of kidney
weight in the control group and decrease of the same
in the treated groups was significant (F = 2.045:
P<0.025). This trend was also reflected in the
renosomatic indices (F= 2.822: P<0.002; Table 1).

3.2. Histopathology of liver

In untreated control group, the transverse section of
liver showed usual structures of the hepatocytes with
normal central vein, Kupffer cells and sinusoids (Plate
1.C). Whereas at lower (0.5-1.0 mg/kg) and higher
(1.5-2.0 mg/kg) doses of Carbofuran-treated diet for
15, 25, 35 and 45 days, the following
histopathological abnormalities were recorded
(summarized in Table 2):
15 days post-treatment: Shrinkage of portal vein
and haemosiderin in Kupffer cells (Plate 1.1);
binucleated hepatocytes, haemosiderin in hepatocytes,
fibrous central and portal veins (Plate 1.2);
haemosiderin in epithelial cells (Plate 1.3); and
haemorrhage and dilated sinusoids (Plate 1.4) were
salient manifestations.
25 days post-treatment: Haemorrhage
accompanied by cell debris, vacuolated and congested
central vein (Plate 2.1); haemorrhage and dilated
sinusoid (Plate 2.2); congested portal vein, cell debris
and vacuolation (Plate 2.3); and congested area and
dilated central vein (Plate 2.4) were common features.
35 days post-treatment: In addition to some of the
aforesaid abnormalities, necrotic hepatocytes and
congested portal vein (Plate 3.1); fibre deposited
central vein (Plate 3.2); inflammatory cell infiltration
and congested central vein (Plate 3.3); and fibrous
tissue accompanied by inflammatory cell infiltration
(Plate 3.4) were manifested.
45 days post-treatment: The most drastic changes
in liver cells following a prolonged Carbofuran
treatment included haemorrhage and inflammatory
cell infiltration (Plate 4.1); increased number of
mononuclear cells and dilated portal vein (Plate 4.2);
fibre deposited portal vein accompanied by cell debris
and haemorrhages (Plate 4.3); and enucleated cells,
necrotic hepatocytes and increased number of
mononuclear cells (Plate 4.4).

Islam et al.
Carbofuran-Induced Alterations in Body Morphometrics and Histopathology of Liver and Kidneys in the Swiss Albino
Mice Mus Musculus L.
311











Table 1: Effect of Carbofuran treated diet on the relative body, liver and kidney weights (g) of the male Swiss albino mice
Parameters/
Treatments
Initial (0 day)

15 days

25 days


35 days


45 days


F-values
(Probabilities)
Body weight


2.668
(0.004)
T0 21.681.50
a
22.931.55
a
24.171.65
a

25.501.84
a

28.10
T1 21.332.37
a
19.882.73
c
18.402.84
b

15.700.28
c

15.00
T2 21.203.08
a
19.552.80
c
19.171.96
b

16.351.63c
15.20
T3 20.503.12
a
20.233.14
b
20.032.28
b

18.402.26
b

18.10
T4 21.582.55
a
19.202.64
c
17.972.15
b

16.201.70
c

15.20
Liver weight
(Hepatosomatic index)



3.759 (0.001)
(1.750; ns)
T0
1.580.17
a

(7.300.94)
1.850.21
a

(8.08084)
1.670.21
a

(6.951.29)
1.600.01
a

(6.290.45)
1.60
(5.69)

T1
1.400.22
b

(6.550.37)
1.350.31
b

(6.841.69)
1.070.15
c

(5.840.85)
1.200.01
b

(7.650.13)
1.20
(8.00)

T2
1.380.22
b

(6.581.38)
1.380.37
b

(7.031.62)
1.230.15
b

(6.491.09)
1.200.00
b

(7.380.73)
1.00
(6.58)

T3
1.330.22
b

(6.470.47)
1.130.19
c

(5.590.64)
1.330.06
b

(6.700.71)
1.150.07
b

(6.280.39)
1.10
(6.08)

T4
1.330.19
b

(6.160.39)
1.100.22
c

(5.710.69)
1.00020
c

(5.530.46)
0.950.21
c

(5.830.70)
0.80
(5.26)

Kidney weight
(Renosomatic index)



2.045 (0.025)
(2.822; 0.002)
T0
0.550.03
a

(2.520.08)
0.550.05
a

(2.390.05)
0.570.02
a

(2.350.07)
0.550.04
a

(2.160.01)
0.62
(2.21)

T1
0.480.02
b

(2.260.24)
0.520.03
a

(2.610.18)
0.500.03
b

(2.760.24)
0.470.01
b

(3.000.04)
0.46
(3.07)

T2
0.530.03
a

(2.530.44)
0.510.03
a

(2.610.24)
0.530.01
b

(2.800.24)
0.490.01
b

(3.010.21)
0.48
(3.16)

T3
0.560.02
a

(2.760.43)
0.540.02
a

(2.710.34)
0.520.05
b

(2.600.05)
0.500.06
b

(2.720.03)
0.54
(2.98)

T4
0.540.03
a

(2.500.26)
0.510.03
a

(2.650.19)
0.530.04
b

(2.940.13)
0.490.04
b

(3.030.06)
0.48
(3.16)




Values are mean SD; dissimilar superscript letters for a parameter or treatment in the same column differ significantly by (LSD) tests (P<0.05); ns= not
significant.


International Journal of Scientific Research in Environmental Sciences, 2(9), pp. 308-322, 2014
312

Plate 1: Transverse sections of liver of Carbofuran treated mice after 15 days. Slides for the control (T0) and treatment groups
(T1-T4) are designated by 1.C, 1.1, 1.2, 1.3 and 1.4, respectively. Abbreviations: BD= bile duct; BH= bi-nucleated
hepatocytes; CV= congested vein; DS= dilated sinusoid; FCV= fibre-deposited central vein; FPV= fibre-deposited portal vein;
H= hepatocytes; HA= hepatic artery; HE= haemorrhage; HmE= haemosiderin epithelial cells; HmH= haemosiderin
hepatocytes; HmK= haemosiderin Kupffer cells; K= Kupffer cells; PV= portal vein; SPV= shrunken portal vein.
Islam et al.
Carbofuran-Induced Alterations in Body Morphometrics and Histopathology of Liver and Kidneys in the Swiss Albino
Mice Mus Musculus L.
313


Plate 2: Transverse sections of liver of Carbofuran treated mice after 25 days. Slides for the treatment groups (T1-T4) are
designated by 2.1, 2.2, 2.3 and 2.4, respectively. Abbreviations: BH= bi-nucleated hepatocytes; CA= congested area; CBD=
congested bile duct; CCV= congested central vein; CDB= cell debris; CPV= congested portal vein; CV=; congested vein;
DCV= dilated central vein; DS= dilated sinusoid; HE= haemorrhage; HmH= haemosiderin hepatocytes; HmK= haemosiderin
Kupffer cells; PV= portal vein.


Plate 3: Transverse sections of liver of Carbofuran treated mice after 35 days. Slides for the treatment groups (T1-T4) are
designated by 3.1, 3.2, 3.3 and 3.4, respectively. Abbreviations: BD= bile duct; BH= bi-nucleated hepatocytes; CBD=
congested bile duct; CCV= congested central vein; CPV= congested portal vein; CV= central vein; DS= dilated sinusoid; F=
fibrous tissue; FCV= fibre-deposited central vein; HE= haemorrhage; HmE= haemosiderin epithelial cells; HmK=
haemosiderin Kupffer cells; INF= inflammatory cell infiltration; NH= necrotic hepatocytes; PV= portal vein; V= vacuolation.
International Journal of Scientific Research in Environmental Sciences, 2(9), pp. 308-322, 2014
314


Plate 4: Transverse sections of liver of Carbofuran treated mice after 45 days. Slides for the treatment groups (T1-T4) are
designated by 4.1, 4.2, 4.3 and 4.4, respectively. Abbreviations: BD= buile duct; CBD= congested bile duct; CDB= cell
debris; CV= central vein; DPV= dilated portal vein; DS= dilated sinusoid; EC= enucleated cells; FPV= fibre-deposited portal
vein; HE= haemorrhage; HmH= haemosiderin hepatocytes; HmK= haemosiderin Kupffer cells; IMC= increased number of
mononuclear cells; INF= inflmmatory cell infiltration.

Table 2: Major histopathological changes detected before and after Carbofuran treatments in the Swiss albino mice
Treatment groups/
Organs
Major histopathological changes

Severity
1

Liver
T0 Usual structures of the hepatocytes with normal central vein, Kupffer cells and sinusoids _

T1 Shrunken portal vein and haemosiderin in Kupffer cells; binucleated hepatocytes, haemosiderin in
hepatocytes, fibrous central and portal veins; haemosiderin in epithelial cells; haemorrhage and
dilated sinusoids

+
T2 Haemorrhage accompanied by cell debris, vacuolated and congested central vein; haemorrhage and
dilated sinusoid; congested portal vein, cell debris and vacuolation; and congested area and dilated
central vein

++
T3 Necrotic hepatocytes and congested portal vein; fibre deposited central vein; inflammatory cell
infiltration and congested central vein; and fibrous tissue accompanied by inflammatory cell
infiltration

+++
T4 Haemorrhage and inflammatory cell infiltration; increased number of mononuclear cells and dilated
portal vein; fibre-deposited portal vein accompanied by cell debris and haemorrhages; and
enucleated cells, necrotic hepatocytes and increased number of mononuclear cells

++++
Kidney
T0 Normal arrangements of Bowmans capsule, glomerulus, urinary pulp, distal and proximal
convoluted tubules and podocytes
_
T1 Oedema and congested glomerulus accompanied by widened urinary space; inflammatory cell
infiltration and vacuolated tubules; dilated collecting duct, medullary ray and distal tubule, increased
number of mesangial cells and degenerated glomerulus

+
T2 Dilated collecting ducts, vacuolated tubules, cell debris, congested glomerulus and widened urinary
space; shrunken glomeruli, presence of inflammatory cell infiltration and hyalinized area; dilated
tubule, eroded wall of Bowmans capsule and increased number of podocytes

++
T3 Haemorrhage accompanied by degenerated or congested glomerulus and widened urinary space;
dilated proximal tubule and increased number of mesangial cells; inflammatory cell infiltration and
dilated tubule; and vacuolated tubule and degenerated Bowmans capsule

+++
T4 Vacuolated distal tubule and haemorrhage; dilated collecting and proximal tubules and degenerated
glomerulus; increased number of podocytes, large mononuclear cells and eroded wall of Bowmans
capsule; and shrinkage of proximal tubule, increased number of podocytes and haemorrhage

++++
1
_ = no change; += minor; ++=not severe; +++= severe; ++++=very severe.

Islam et al.
Carbofuran-Induced Alterations in Body Morphometrics and Histopathology of Liver and Kidneys in the Swiss Albino
Mice Mus Musculus L.
315
3.3. Histopathology of kidney

The transverse sections of untreated control renal cells
revealed normal arrangements of Bowmans capsule,
glomerulus, urinary pulp, distal and proximal
convoluted tubules and podocytes (Plate 5.C). In
contrast, the Carbofuran-treated feed induced the
following exposure-dependent symptoms in the
experimental mice (summarized in Table 2):
15 days post-treatment: At lower doses oedema
and congested glomerulus (Plate 5.1); congested
glomerulus accompanied by widened urinary space
(Plate 5.2); while a higher doses inflammatory cell
infiltration and vacuolated tubules (Plate 5.3); and
dilated collecting duct, medullary ray and distal
tubule, increased number of mesangial cells and
degenerated glomerulus (Plate 5.4) were common
features.
25 days post-treatment: Such symptoms as dilated
collecting ducts, vacuolated tubules, cell debris,
congested glomerulus and widened urinary space
(Plate 6.1); shrinkage of glomerulus, inflammatory
cell infiltration and hyalinized area (Plate 6.2); dilated
tubule, eroded wall of Bowmans capsule and
increased number of podocytes (Plate 6.4) were
manifested.
35 days post-treatment: Haemorrhage
accompanied by degenerated or congested glomerulus
and widened urinary space (Plate 7.1); dilated
proximal tubule and increased number of mesangial
cells (Plate 7.2); inflammatory cell infiltration and
dilated tubule (Plate 7.3); and vacuolated tubule and
degenerated Bowmans capsule (Plate 7.4) were
salient features.
45 days post-treatment: In addition to some of the
aforesaid anomalies, vacuolated distal tubule and
haemorrhage (Plate 8.1); dilated collecting and
proximal tubules and degenerated glomerulus (Plate
8.2); increased number of podocytes, large
mononuclear cells and eroded wall of Bowmans
capsule (Plate 8.3); and shrinkage of proximal tubule,
increased number of podocytes and haemorrhage
(Plate 8.4) were notable characteristics.

4. DISCUSSION

In this investigation, a detailed account of Carbofuran-
fed changes in body, liver and kidney weights,
coupled with corresponding toxic effects of the
carbamate insecticide in the liver and kidney tissues of
male Swiss albino mice has been presented. The
results of the current study revealed that there were
significant decreases in body and liver weights in
mice treated with Carbofuran. In toxicological studies,
however, organ and relative organ weights are
important criteria for evaluation of organ toxicity
(Crissman et al., 2004). The organ and relative organ
weights may be increased or decreased depending on
the nature and mode of action of the insecticide.
However, the present data corroborate to Pant et al.
(1995) and El-Damaty et al. (2012) but differ from
Brkic et al. (2008) where a statistically significant
increase in body weight of male rats treated with
Carbofuran at 400 mg/kg body weight was observed.
In an earlier study, Khogali et al. (2005) reported
Dimethoate-induced hepatic pycnosis, vacuolation,
blood congestion and high lymphatic infiltration
around the central vein; and changes in the cortex at
the glomeruli as swollen cellular lining of the
Bowmans capsule in Swiss albino mice. While oral
administration of Carbosulfan at 48 mg/kg/day for 5,
10, 20 and 30 days in albino mice resulted in dilation
of central vein and sinusoids between hypertrophied
hepatocytes, vacuolization and hyalinization of
hepatocytes with loss of radial arrangement,
suggesting that the insecticide had adverse effects on
liver functions leading to histological and
physiological impairments (Ksheerasagar and
Kaliwal, 2006). Further Ksheerasagar and Kaliwal
(2010) administered Carbosulfan at 12, 24 mg/kg/day
orally for 30 days in mice and noticed in the loss of
normal arrangement of cortical tubules and formation
of vacuoles in hepatic tissues; while Carbosulfan at 36
and 48 mg/kg/day for 30 days resulted in atrophied
glomeruli which were loosely attached to Bowmans
capsules. Owing to a prolonged exposure of 45 days,
however, the present results appeared to be more
drastic on mice organs than those discussed above.
In Asian common toad tadpoles, Duttaphrynus
melanostictus exposure of 50-500 g/L Carbofuran
for 15 days resulted in greater vacuolation in
hepatocytes, sinusoidal dilations and the formation of
bile plugs (Jayatillake et al., 2011). El-Damaty et al.
(2012) examined the effects of Carbofuran on liver
and kidney function parameters in male albino rats, in
which 1/10
th
of the LD
50
dose resulted in the decrease
of body weight but increased the liver and kidney
weights. Liver histopathology at this dose included
pyknotic nuclei, focal necrosis with inflammatory
infiltration, vacuolization and blood congestion;
whereas kidney histopathology included blood
congestion in between tubules and small area of
haemorrhage in the interstitial tissues. These findings
are slightly different from those of the present
observations perhaps because of different dose levels
and exposure durations.

International Journal of Scientific Research in Environmental Sciences, 2(9), pp. 308-322, 2014
316

Plate 5: Transverse sections of kidney of Carbofuran treated mice after 15 days. Slides for the control (T0) and treatment
groups (T1-T4) are designated by 5.C, 5.1, 5.2, 5.3 and 5.4, respectively. Abbreviations: BC= Bowmans capsule; CG=
congested glomeruli; DCD= dilated collecting duct; DDT= dilated distal tubule; DG= degenerated glomeruli; DMR= dilated
medullary ray; DT= dilated tubule; EWB= eroded wall of Bowmans capsule; G= glomerulus; IPC= increased number of
podocytes; IMC= increased number of mesangial cells; OD= oedema; PC= podocytes; PT= proximal convoluted tubule; UP=
urinary palp; VT= vacuolated tubule; WUS= widened urinary space.

Islam et al.
Carbofuran-Induced Alterations in Body Morphometrics and Histopathology of Liver and Kidneys in the Swiss Albino
Mice Mus Musculus L.
317

Plate 6: Transverse sections of kidney of Carbofuran treated mice after 25 days. Slides for the treatment groups (T1-T4) are
designated by 6.1, 6.2, 6.3 and 6.4, respectively. Abbreviations: CDB= cell debris; CG= congested glomeruli; DCD= dilated
collecting duct; DG= degenerated glomeruli; DMR= dilated medullary ray; DT= dilated tubule; EWB= eroded wall of
Bowmans capsule; G= glomerulus; HA= hyalinized area; INF= inflammatory cell infiltration; IPC= increased number of
podocytes; OD= oedema; PT= proximal convoluted tubule; SG= shrinked glomeruli; VT= vacuolated tubule; WUS= widened
urinary space.


Plate 7: Transverse sections of kidney of Carbofuran treated mice after 35 days. Slides for the treatment groups (T1-T4) are
designated by 7.1, 7.2, 7.3 and 7.4, respectively. Abbreviations: CG= congested glomeruli; DBC= degenerative Bowmans
capsule; DCD= dilated collecting duct; DDT= dilated distal tubule; DG= degerated glomeruli; DPT= dilated proximal tubule;
DT= dilated tubule; G= glomeruli; HE= haemorrhage; IMC= increased number of mesengial cells; INF= inflammatory cell
infiltration; PT= proximal convoluted tubule; VT= vacuolated tubule; WUS= widened urinary space.

International Journal of Scientific Research in Environmental Sciences, 2(9), pp. 308-322, 2014
318

Plate 8: Transverse sections of kidney of Carbofuran treated mice after 45 days. Slides for the treatment groups (T1-T4) are
designated by 8.1, 8.2, 8.3 and 8.4, respectively. Abbreviations: CG= congested glomeruli; DCD= dilated collecting duct;
DG= degenerative glomeruli; DPT= dilated proximal tubule; DT= dilated tubule; EWB= eroded wall of Bowmans capsule;
HE= haemorrhage; IMC= increased number of mesangial cells; IPC= increased number of podocytes; OD= oedema; PT=
proximal convoluted tubule; SPT= shrinked proximal tubule; VDT= vacuolated distal tubule; WUS= widened urinary space.

Oral administration of Dimethoate in albino mice
at 7, 14 and 28 mg/kg body weight resulted in
abnormal liver and kidney histopathology that
included hepatocytic pycnosis, vacuolization, blood
congestion, high lymphatic infiltration around the
central vein, enlargement of hepatic sinusoids,
hepatocellular damage, necrosis, increase in the
number of Kuffer cells, lesions and hemorrhage
(Yasin and Sharma, 2013). Recently, El-bendary et al.
(2014) evaluated histopathological abnormalities
associated with Profenofos and Chloropyrifos
exposure in male albino mice where liver showed
hepatic cell damage with degenerative changes
including congestion of blood vessels, vacuolar
degeneration of hepatic cells, focal infiltration and
mononuclear cells, dilated central vein and other
hepatic blood vessels, necrosis of hepatic cells,
disorganized with the formation of denoid structure
and hepatocytomelagy; whereas kidney showed
hemorrhage, oedema, necrosis and glomerular
shrinkage. In addition, histopathological abnormalities
in kidney cells included perivascular oedema with
congestion of renal blood vessels, infiltration of
mononuclear cells around glomerular tubules, oedema
of Bowmans capsule and coagulation necrosis of
some renal tubules. These results appear to fit well
with those of the present investigation.
Being a broad spectrum carbamate pesticide,
Carbofuran is extensively used against soil and foliar
pests of field, fruit, vegetables and forest crops, which
has been reported to be highly toxic by inhalation and
ingestion, and moderately toxic by dermal absorption
(WHO, 1985). It is highly toxic to aquatic biota
including fishes, environment as well as humans and
is reported to propagate through oral and inhalation
routes of exposure (Baron, 1991). Moreover,
environmental pollution and health hazards including
cases of severe, sub-severe and chronic human and
wildlife poisoning through food chain or contaminated
foods and drinking water have been reported (Aziz et
al., 2008; Chowdhury et al., 2012). Because humans
are at the top of the food chain, it is probable that
detrimental residues of Carbofuran may remain in the
edible portion of the vegetable or plants and thus may
affect human health and well-beings. Several reports
demonstrate that exposure of Carbofuran results in
some histopathological changes in liver and kidney of
human and other mammals that include mononuclear
cell infiltration, congestion, enlargement of the veins
and sinusoids, necrosis, increased number of Kupffer
cells, cytoplasmic vacuolation and degenerative
hepatocytes (Aziz et al., 2008; El-Damaty et al.,
2012). These are in good agreement with the present
results on histopathological changes in the
experimental mice.
Islam et al.
Carbofuran-Induced Alterations in Body Morphometrics and Histopathology of Liver and Kidneys in the Swiss Albino
Mice Mus Musculus L.
319
Recent studies showed that indiscriminate and
unregulated uses of insecticides like Carbofuran in
agriculture and public health in Bangladesh have led
to drastic effects in many non-target species including
man (Chowdhury et al., 2012; Uddin, 2013). Since the
present feed treatments of albino mice with
Carbofuran clearly demonstrated decrease in body,
liver and kidney weights as well as marked changes in
the histopathology of hepatic and renal tissues, it
could be possible that prolonged exposure to this
insecticide in man may play a significant role in
aggravating such diseases as chronic liver and renal
failure. In addition, structural changes to hepatic and
renal tissues such as haemorrhage, congestion,
vacuolation and erosion may also lead to acute liver or
kidney damages and/or carcinogenicity of the organs.
The present results could thus be exploited as a
potential biomarker of common insecticide toxicity in
human beings.

5. CONCLUSION

The present findings clearly demonstrate that
Carbofuran is capable of inducing dose-dependent
morphometric i.e. body, liver and kidney weights and
their relative weights as well as histopathological
changes in the liver and kidney of the exposed mice.
According to these data, it is suggested that systemic
insecticide like Carbofuran exposure might cause
hazardous effects, especially at high doses, to man and
environment. For field and domestic uses of this
insecticide, quantities and mode of usage need to be
strictly monitored to minimize the possibility of its
exposure to non-target organisms including human
beings. This can be achieved through public health
education to make people aware of the hazardous
effects of this chemical. It is therefore recommended
that great precautions are to be taken to minimize the
harmful side effects of Carbofuran to the environment,
especially to man, animals and agricultural products,
aiming at avoiding environmental pollution.
Moreover, recommended doses of the insecticide and
precautionary measures like wearing of impermeable
gloves and masks to reduce the risk of inhalation of
spray should be implemented. Due attention also is to
be paid for a delayed period of field application of this
insecticide to avoid its possible adverse effects to
consumers, who should be warned of the potential risk
of Carbofuran contamination of food and drinking
water in the country.

Acknowledgements

The authors are grateful to the Chairpersons,
Departments of Zoology and Genetic Engineering &
Biotechnology, University of Rajshahi, Bangladesh,
for providing laboratory facilities, and to Laboratory
Attendants, for their technical assistance. Sincere
thanks must go to the anonymous reviewer for his
useful comments and suggestions.

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321

Dr. M. Saiful Islam, Professor, Department of Zoology, University of Rajshahi, Rajshhai 6205,
Bangladesh. The author was born on December 4, 1955 at Bogra, Bangladesh. He received his BSc
Honours (1977) and MSc (1978) Degrees in Zoology from the University of Rajshahi, Bangladesh,
where he started his teaching and research career as a Lecturer in Zoology in 1983. Subsequently he had
his second MSc in Applied Entomology and Crop Protection from the University of Newcastle upon
Tyne, UK (1990), PhD in Insect Physiology and Behaviour from the Universities of Reading and
Oxford, UK (1995), Commonwealth Academic Fellowship in the Department of Zoology, University of
Oxford, UK (2001-2002) and Visiting Fellowship in the Department of Entomology, University of
Kentucky, USA (2003-2005). Meanwhile, Dr. Islam was promoted to Assistant Professor (1986),
Associate Professor (1995) and Professor (1998) in his parent Department, where he is currently
employed. Professor Islam has supervised quite a good number of MSc thesis (>22) and PhD students (17) since 1983. He has
a credit of publishing over 120 research papers in renowned scientific journals of home and abroad. In addition, he co-authored
a couple of text books titled Biotechnology and Genetic Engineering and Introduction to Genetics while his another book
named Genetics: The Science of Heredity and Variation is awaiting publication. Prof. Islam is a member of a large number
learned bodies: Fellow of the Royal Entomological Society (FRES), London; Fellow of the Zoological Society of Bangladesh
(ZSB); Life Member, Zoological Society of Bangladesh (Regional Secretary, 1996-1998); Life Member, Genetical Society of
Bangladesh (Treasurer, 1999-2005); Life Member, Bangla Academy, Bangladesh; Life Member, Association of the
Bangladesh Commonwealth Scholars; Life Member, Bangladesh Entomological Society; Member, Bangladesh Association for
the Advancement of Science (BAAS); Member, Asiatic Society of Bangladesh and Ex-Member, American Association for the
Advancement of Science (AAAS). He has conducted research for a number of grants funded by various organizations
including the University of Rajshahi, Ministry of S & T, UGC of Bangladesh, TWAS (Italy) and IAEA (Austria). Prof. Islam
served as an Executive Editor and Editorial Board member of several scientific journals and periodicals. He teaches vertebrate
and invertebrate Zoology, Microbiology, Biostatistics, Evolution, Embryology & Developmental Biology and Animal
Behaviour to undergraduate, and Genetics & Molecular Biology to postgraduate students. His current research interest
includes: Genetics and Molecular Biology of pest insects, inheritance of Mendelian and non-Mendelian traits in man, and
applications of breeding principles to farm animals. Email address: saifulzoo@yahoo.co.uk.



Dr. Moni Krishno Mohanta, Assistant Professor, Department of Zoology, University of Rajshahi,
Rajshahi 6205, Bangladesh. Dr. Mohanta was born on March 4, 1979. He received his BSc in
Zoology (2001), MSc in Zoology (Genetics and Molecular Biology) in 2002 from the University of
Rajshahi, Bangladesh and PhD in Environmental Microbiology from the same University in 2009.
Presently he is working as an Assistant Professor in the Department of Zoology, Rajshahi University,
Bangladesh. He has published a dozen of papers in different Journals. So far Dr. Mohanta has
supervised eight research students at BSc and MSc levels. He teaches Cell Biology, Paleontology and
Reproductive Biology to undergraduate and Microbial Genetics and Biotechnology to postgraduate
students. His current research interest lies in the areas of Environmental Microbiology and
Toxicology. Email: mkmohanta_zool@yahoo.com



Dr. Ananda Kumar Saha, Professor, Department of Zoology, University of Rajshahi, Rajshhai 6205,
Bangladesh. Dr. Saha was born on October 31, 1957 at Magura, Bangladesh. He received his BSc
Honours (1979) and MSc (1980) Degrees in Zoology from the University of Rajshahi, Bangladesh,
where he started his teaching and research career as a Lecturer in Zoology in 1987. Subsequently he
had his second MSc in Environmental Microbiology from the University of Newcastle upon Tyne,
UK (1991), PhD in Microbiology from the University of Pune, India (2002). Presently he is working
as a Professor in the Department of Zoology, Rajshahi University, Bangladesh. Professor Saha has
supervised quite a good number of MSc thesis and PhD students. He has published over 40 research
papers in renowned scientific journals of home and abroad. Prof. Saha is a member of a large number
learned bodies: Life Member, Zoological Society of Bangladesh (ZSB), Life Member, Genetical
Society of Bangladesh, Life Member, National Environmental Science Academy, India, Life Member, Biodiversity Research
Group of Bangladesh (BRGB), Life Member, Microbiological Society of Bangladesh, Member, Bangladesh Association for
the Advancement of Science (BAAS), Member, Asiatic Society of Bangladesh and Member, Microbiological Society of India.
He teaches Cell Biology and Microbiology to undergraduate and Genetics & Molecular Biology to postgraduate students. His
current research interest lies in the areas of Environmental Microbiology, Waste water management and Toxicology. Email
address: anandroma@yahoo.com.



International Journal of Scientific Research in Environmental Sciences, 2(9), pp. 308-322, 2014
322



Anup Mondol, Microbiologist, Mostofa Organic Shrimp Product Ltd., Satkhira, Khulna. Born on the
15
th
January, 1988, Gopalgonj, Bangladesh. Mr. Mondol received his BSc Honours (2009) and MSc
(2010) degrees in Zoology from University of Rajshahi, Bangladesh, in which he got First Class in
both examinations. He has two years research experience during which he conducted his
undergraduate and graduate studies. He has developed good computer skills and has commendable
language proficiencies in Bengali, English. Email: anupmondolzool@gmail.com






Muhammad Mizanul Hoque, Assistant Officer, Islami Bank Bangladesh Limited, Natore. Born on the
2
nd
of August, 1985, Mr. M. M. Hoque received his BSc Honours in Biotechnology & Genetic
Engineering from Khulna University, Bangladesh in 2008. His masters thesis on Swiss albino mice
in the Department of Zoology, Rajshahi University, was highly commendable and he was awarded
MS from his parent department in 2013. Owing to his extraordinary performance Mr. Hoque was
awarded Merit Scholarships three times during the academic sessions of 2005-2006, 2007-2008 and
2009-2010. He is currently employed by the Islami Bank Bangladesh Limited where he is an
Assistant Officer in ICT-based banking operations. Email: mhoque.bge@gmail.com.





Dr. Apurba Kumar Roy, Associate Professor, Department of Genetic Engineering and Biotechnology,
University of Rajshahi, Rajshahi 6205, Bangladesh. Dr. Roy was born on the 17
th
May, 1970 at
Narail, Bangladesh. He received his BSc (Honours) degree (1992) from the Department of Botany,
Rajshahi University, Bangladesh. He received his MSc (1994) in the Department of Genetics and
Breeding and PhD in Genetics in the same University where he started his research and teaching
career as Lecturer. Currently he is an Associate Professor in the Department of Genetic Engineering
and Biotechnology. He teaches Fundamentals of Botany, Developmental Biology, Pathology, Human
Genetics, Immunogenetics, Food Biotechnology and Biomedical Science to undergraduate and
postgraduate students. His current research interest lies in the areas of Biomedical Science and
Toxicology. Email: apu_gen@yahoo.com.