Indian J. Vet. Pathol., 38(3): 186-189, 2014; DOI: 10.5958/0973-970X.2014.01169.

Pathological changes induced by acephate and its amelioration

with vitamin E in broiler chicken
R.K. Pradip, A. Anand Kumar* and M. Lakshman
College of Veterinary Science, Rajendranagar, Hyderabad-500030
Received: 22.02.14; Accepted: 10.07.14

ABSTRACT
Pradip, R.K., Kumar, A.A. and Lakshman, M. (2014). Pathological changes induced by acephate and its amelioration with vitamin E in
broiler chicken. Indian J. Vet. Pathol., 38(3) : 186-189.

Acephate is one of the top ten organophosphate insecticides sold throughout the world. The present study was undertaken to assess
the toxic effects of acephate and its amelioration, if any, with vitamin E. Ninety broiler chicks were divided into six groups of 15 birds in each
and fed with acephate at the dose rates of 85.2 mg/kg and 170.4 mg/Kg feed for six weeks. In acephate-fed groups, gross pathological
changes observed were moderate enlargement and paleness of liver, swollen and hemorrhagic kidneys, petechiae on pericardium, and mild
congestion of brain. Histological changes comprised of fatty change in liver and kidney, degeneration and necrosis of hepatocytes and
tubular epithelium, myocardial degeneration with infiltration of mononuclear cells in muscle fibres, depletion of lymphocytes in spleen and
cystic spaces in bursal follicles, with increased severity in higher dose group and age advancement. Ameliorative groups did not reveal any
gross lesion and showed mild to moderate improvement in histological lesions. It was concluded that Acephate, fed at above levels, results
in multiple organ toxicity. Supplementation of vitamin E at 300 mg/Kg feed ameliorated the toxic effects produced by acephate.
Downloaded From IP - 117.211.167.37 on dated 1-Jun-2018

Keywords: Acephate, broiler chicken, pathology, vitamin E

Members Copy, Not for Commercial Sale
www.IndianJournals.com

INTRODUCTION decrease viability of embryos, chicks and reduction in

Application of pesticides in agriculture, horticulture
and forestry may result in environmental
contamination. Pesticides together with heavy metal
emissions are dominant compounds in the chemical load
of the environment of man and animals1. The United
States Environmental Protection Agency (USEPA)
defines pesticide as any substance or mixture of
substances intended for preventing, destroying,
repelling or mitigating any pest. Besides their great
contribution to agricultural practice, animal and human
populations for prevention of vectors of diseases,
extensive use of pesticides creates many problems viz.,
their toxicity for non-target organisms, residual
persistence in environment and combined effects with
other agrochemicals and environmental factors etc. have
been reported worldwide2.
Acephate is one of the top ten organophosphate
insecticides sold throughout the world. Methamidophos
is the primary metabolite of acephate in plants, birds
and mammals, is a significantly more potent inhibitor
of cholinesterase activity than Acephate. As acephate
insecticide is used as crop protectants they are likely to
cause indirect exposure in poultry through feed
contaminants, soil and ground water and may affect
the biological systems of birds. Field studies and
incidents indicate that the use of acephate may have a
detrimental effect on birds. Chronic exposure of acephate
causes detrimental effects on avian reproduction,
*Corresponding author: e-mail: aakumar7@rediffmail.com
number of egg production3. Free radical production
occurs continuously in all the cells as a by-product of
cellular metabolism. Acephate generated free radicals
induce oxidative damage in biological system. Oxidative
damage by free radicals can be prevented by the use of
antioxidants such as ascorbic acid, tocopherols,
carotenoids and certain herbs4.
MATERIALS AND METHODS
Experimentation
Experiment was conducted on day old broiler chicks
of Vencobb strain, procured from Venkateswara
Hatcheries, Hyderabad. The study was conducted on 90
broiler chicks divided into six groups with 15 birds in
each to assess the toxic effect of acephate (Acephate 75%
SP, Mfg. By: Coromondal International Ltd., Vellore,
Tamilnadu) and its amelioration with vitamin E (DL-
alpha tocopherol acetate; Himedia, Mumbai). The
experiment was conducted for a period of 42 days and
birds were slaughtered at 14th, 28th and 42nd day with
prior approval of Institutional Animal Ethics Committee.
Apparent LD50 of acephate (852 mg/kg) was taken into
consideration for calculation of different dose groups5.
Group I: Birds was maintained on basal feed for 6 wks.
Group II: Birds received acephate at the rate of 85.2 mg/
kg feed for six weeks.
Group III: Birds received acephate at the rate of 170.4
mg/Kg feed for six weeks.
 Pathological changes induced by acephate and its amelioration with vitamin E
Group IV: Birds received acephate at the rate 85.2 mg/
kg feed and vitamin E 300 mg/Kg feed.
Group V: Birds received acephate at the rate 170.4 mg/
Kg feed and vitamin E 300 mg/kg feed.
Group VI: Birds received vitamin E at the rate of 300
mg/kg feed.
Pathology
The birds were sacrificed as per the schedule given
above and gross pathological changes, if any, were noted.
Representative tissue pieces from heart, liver, kidney,
spleen, bursa and brain were fixed in 10% neutral
buffered formalin for histopathological studies. Formalin
fixed tissues were processed routinely to obtain 5 µm
thick paraffin-embedded sections and stained with
routine Hematoxylin and Eosin (H&E) for microscopic
examination6.
RESULTS
Downloaded From IP - 117.211.167.37 on dated 1-Jun-2018
Gross pathology
Groups I and VI: No lesions of pathological significance
Members Copy, Not for Commercial Sale
were observed in heart, liver, kidneys, spleen, bursa and
www.IndianJournals.com
brain at different intervals.
Groups II and III:
Heart: Petechial haemorrhages were recorded on
pericardium at 4 th week. No gross changes were
observed at 2nd and 6th week in both the groups.
Liver: At the end of 2nd, 4th and 6th week, there was mild to
moderate enlargement of liver with pale discoloration
and friable consistency in group II. In group III, the gross
changes were relatively severe and liver was pale and
enlarged and turgid with round edges, and had friable
consistency at 2 nd, 4 th and 6 th week and petechial
haemorrhage was noted at 6th week (Fig. 1).
Kidney: Kidneys of both the groups did not show any
lesions of pathological significance at 2nd week. From 4th
week onwards, little swelling was observed in group II
and congestion and haemorrhages were noted in group
III.
Spleen: No pathological changes were recorded at the end
of 2nd week. Mild to moderate congestion and reduction
in size was recorded at 4th and 6th week in both the
groups.
Brain: Mild congestion was noticed at 4th week. No gross
changes were observed at 2nd and 6th week in both the
groups.
Bursa of Fabricius: No lesions of pathological significance
were observed in both the groups.
Groups IV and V: Liver was friable in consistency at
the end of 2nd week and appeared slightly enlarged at 4th
and 6th week in both the groups. Kidneys revealed mild
187
to moderate congestion at 6th week in both the groups,
whereas heart, spleen, bursa of fabricius and brain did
not show any significant lesion.
Histopathology
Heart: No histological changes were noted in groups I, II,
IV and VI. Mild myocardial degeneration was observed
in groups III and V at 2 nd week. At 4 th week: No
histological changes were detected in groups I and VI,
however, group II, IV and V birds revealed mild
degenerative changes in endocardium and myocardium
accompanied with extravasation of erythrocytes. Group
III birds showed mild to moderate degenerative changes
in myocardium, focal infiltration of mononuclear cell
and separation of muscle fibres (Fig. 2). At 6th week: No
histological changes were detected in groups I and VI.
Myocardial haemorrhages, moderate degeneration and
separation of muscle fibres were observed in group II.
Severe myocardial degeneration and infiltration of
mononuclear cells was observed in group III. Mild
degenerative changes and separation of muscle fibres in
myocardium were observed at the end of 6th week in
groups IV and V.
Liver: No histological changes were detected in any group
at 2nd week. At 4th week: Group II birds showed moderate
to severe fatty changes, mild to moderate degeneration
and necrosis of hepatocytes and focal mononuclear cell
infiltration. In group III, these changes were relatively
severe. Microscopic changes of hepatocytic degeneration
and necrosis accompanied with mild mononuclear cell
infiltration, sinusoidal dilation and congestion and
lymphoid aggregation in few areas were observed in
group IV and V birds. No histological changes were
detected in group I and VI. At 6th week: Moderate fatty
changes, degeneration and necrosis of hepatocytes,
lymphoid aggregation and focal mononuclear cell
infiltration were observed in group II. These changes
were more marked and of severe intensity in group III
(Fig. 3). Sinusoidal dilatation and mild congestion was
noticed in group IV and V, whereas no microscopic
change was detected in group I and VI.
Kidneys: At 2nd week, no histological changes were
detected in group I, IV and VI, whereas only congestion
and mild degenerative changes in tubular epithelium
was detected in group II, III and V. At 4th week: Moderate
to severe fatty changes, mononuclear cell infiltration and
degeneration of tubular epithelium were observed in
group II, and these changes became more marked and
intense in group III (Fig. 4). In group V, tubular epithelium
showed mild degenerative changes and no changes were
noticed in groups I, IV and VI. At 6th week: Group II birds
evinced mild to moderate degenerative changes in
tubular epithelium, which increased in severity in group
III birds and associated with haemorrhages. Very mild
 188 Pradip et al.
Downloaded From IP - 117.211.167.37 on dated 1-Jun-2018
Members Copy, Not for Commercial Sale
www.IndianJournals.com

Fig. 1. Gross appearance of liver showing increase in size with rounded edges at 6th week in group II and III; Fig. 2. Heart showing separation
of muscle fibres and mononuclear cell infiltration in group III at 4th week (H&E x400); Fig. 3. Liver cells showing severe fatty changes in
group III at 6th week (H&E x400); Fig. 4. Kidneys showing degenerative changes including fatty changes in tubular epithelium in group III
at 4th week (H&E x400); Fig. 5. Bursa showing depletion of lymphocytes and thickened interfolicular septa in group III at 6th week (H&E
x100).
 Pathological changes induced by acephate and its amelioration with vitamin E
degenerative changes in groups IV and V, and no
histological changes were detected in groups I and VI.
Spleen: No histological changes were notice in group I,
IV, V and VI, whereas mild depletion of lymphocytes
occurred in groups II and III at 2nd week. At 4th and 6th
week: No histological changes were detected in groups I
and VI. Mild lymphocytic depletion and mild thickening
of splenic arteries was seen in group II, III, IV and V,
which was associated with intrafollicular vacuolation
in the former two groups. A
Bursa of Fabricius: Bursa of Fabricious exhibited follicular
haemorrhages and depletion of lymphocytes in group II
and severe depletion with atrophied follicles, formation
of cystic spaces, loss of interfollicular septa and
coalescing of follicles was observed in group III at 4th
week. Group IV and V revealed mild to moderate
depletion of follicular lymphocytes. At 6 th week:
Follicular atrophy and depletion of lymphocytes was
observed in group II and III associated with thickened
Downloaded From IP - 117.211.167.37 on dated 1-Jun-2018
interfollicular septa (Fig. 5). Mild to moderate depletion
of lymphocytes was seen in group IV and V, and no
Members Copy, Not for Commercial Sale
lesion was detected in groups I and VI.
www.IndianJournals.com
DISCUSSION
The present study revealed petechial hemorrhages
on pericardium of heart in the Acephate treated groups
at 4th week. Hemorrhages on pericardium might be due
to vascular damage induced by acephate. Acephate
treated groups produced enlarged pale discolored liver,
petechiae with friable consistency with degree of weekly
variability. Swollen, congested and hemorrhagic kidneys
were observed in the acephate treated groups from 4th
week onwards. These findings might be due to
disturbances in the liver and kidney functions induced
by acephate and the similar lesions were observed by
earlier workers7. Congestion and reduction in size of
spleen in the present study might be due to inhibited
hemopoietic activity or extra medullary hemopoietic
activity that was induced by acephate and in other
treated groups revealed mild to moderate lesions. These
findings are in accordance with the reports of earlier
weorkers7-8.
Histological lesions of the acephate treated groups
revealed mild degeneration and presence of hemorrhages
in the endocardium and myocardium with increase in
severity from 4th week onwards might be due to vascular
damage induced by acephate and the severity of lesions
might be due to the prolonged action and different dose
rates of acephate. These findings are in accordance with
the findings of earlier scientists8. Ameliorative groups
showed very mild lesions might be due to protective
189
effect of vitamin E. Acephate treated groups produced
significant histological lesions like hepatocellular
degeneration and necrosis at 4 th and 6 th week that
indicated liver dysfunction and hepatotoxic. These
results are in accordance with earlier reports 7-8.
Ameliorative groups showed reduced severity or
improvement in the condition might be due to protective
effect of vitamin E.
Acephate treated groups produced significant
histological lesions like tubular degeneration and
necrosis at 4th and 6th week and indicated dysfunction of
kidney and nephrotoxic. These changes were also
reported by8, where they indicated that the target organ
for acephate induced toxicity is kidney. Ameliorative
groups showed mild to moderate improvement might
be due to protective effect of vitamin E as it scavenges
free radicals formed.
The present study indicated that acephate at both
the dose rates resulted multiple organ toxicity might be
due to cell damage lead to tissue alterations.
Administration of vitamin E alleviated the toxic effect
produced by acephate due to its antioxidant property.
ACKNOWLEDGEMENTS
The authors are thankful to SV Veterinary
University, Tirupati for providing the facilities to carry
out the present study.
REFERENCES
1. Kacmar P, Pistl J, Mikula I. 1999. Immunotoxicol Vet Med Acta
Veterinaria 68: 57-79.
2. Fleming LE, Bean JA, Rudolph M, Hamilton K. 1999. Mortality
in a cohort of licensed pesticide applicators in Florida. Occup
Environ Med 56: 14-21.
3. EPA (Environmental Protection Agency), 1987. PB 88-179999.
Pesticide Fact Sheet No., 140: Acephate.
4. Kamashi K, Gopala Reddy A, Reddy KS, Reddy VR. 2004.
Evaluation of zinc against salinomycin toxicity in broilers.
Indian J Physiol Pharmacol 48: 89-95.
5. Willcox H, Coffey T. 1977. Environmental Impact of Acephate
(Orthene) on Forest Insect and Disease Management. Depart-
ment of Agriculture Forest Service. Northeastern Area State and
Private Forestry, Upper Darby pp. 8.
6. Luna GLHT. 1968. Manual of histological and special staining
techniques of the Armed Forces Institute of Pathology. 2nd Edn. The
Blakistone Divison McGraw-Hill Book Company, Inc. New
York, Toronto London. pp. 1-5, 9-34.
7. Tripathi SM, Thaker AM, Joshi CG, Garg SP, Snakhla LN.
2007. Immunotoxicity induced by sub-acute acephate exposure
in White Leghorn Cockerels. J Indian Soc Toxicol 3: 13-21.
8. Bhandaniya AR, Joshi DV, Patel BJ, Kalaria VA, Padodara RJ,
Savsani HH. 2012. Toxico-pathological studies on experimen-
tally induced acephate toxicity in Wistar rats (Rattusnorvegicus).
Wayamba J Anim Sci 4: 1-8.