Professional Documents
Culture Documents
ABSTRACT
Babu, N. Sasidhar, Kumar, A. A., Srilatha, Ch., Lakshman, M., Amaravathi, P., Hemanth, I., Sailaja, N. and Sujatha, K. (2015). Histological
and ultrastructural changes induced by long term feeding of imidacloprid and amelioration with vitamin C and Withania somnifera in
layer birds. Indian J. Vet. Pathol., 39(4) : 343-346.
The study was carried out on above 2 months old 48 layer birds for 90 days to elucidate the histopathological and ultrastructural
alterations induced by imidacloprid and to evaluate the protective effect of vitamin C and Withania somnifera. The birds were divided in
Downloaded From IP - 117.211.167.37 on dated 20-Jun-2018
six groups: Group 1 (control), group 2 (imidacloprid @ 200 ppm in feed), group 3 (vitamin C @ 200 ppm in feed), group 4
(imidacloprid@200ppm and vitamin C 200ppm), group 5 (Withania somnifera @ 500 ppm in feed) and group 6 (imidacloprid @ 200 ppm
in feed + Vitamin C @ 200 ppm in feed + Withania somnifera @ 500 ppm in feed).Grossly, congested and enlarged liver and atrophied
Members Copy, Not for Commercial Sale
spleen with varied severity was seen at day 60 and 90 in group 2. Histologically, degenerative and necrotic changes in the hepatic
www.IndianJournals.com
lobules, renal tubular degeneration and atrophied glomeruli andendocardial hemorrhages were observed in group 2. In group 6, mild
to moderate degenerative changes in the hepatic lobules depending on the time interval was noted. Mild pathologicalchanges were
noticed in group 4. Ultrastructurally, cytoplasmic, nuclear, rough endoplasmic reticulum and mitochondria revealed alterationsin liver
and kidney in toxic group. Vitamin C and Withania somnifera supplementation resultedmoderate protection by repair and regeneration
of damaged tissues.
Keywords: Histological, imidacloprid, ultrastructural, vitamin C, Withania somnifera
4
344 Babu et al.
RESULTS
Gross abnormalities observed
in liver were congestion and
enlargement with rounded
boarders and spleen was atrophied
in group 2 and the severity of the
lesions increased as the experiment
progressed (Fig. 1). Histologically,
in imidacloprid group, liver
showed marked dilation of central
vein, engorged sinusoids and
portal vein. Hepatocytes showed
5
Effect of long term feeding of imidacloprid and amelioration with vitamin C and Withania somnifera in birds 345
fatty changes, degeneration, necrosis and Kupffer cell nephrotoxic effects of imidacloprid as kidneys are major
proliferation (Fig.2). In kidney, inter-tubular hemorrhages, excretory organs for many xenobiotics/ intoxicants. These
degeneration of tubular epithelium and shrunken results are in accordance with previous findings in birds9
glomeruli with vacuolation and mesangeal cell and rats 17 . Similarly, observations of endocardial
proliferation were observed (Fig. 3). In heart, endocardial hemorrhages, degeneration and separation of muscle
hemorrhages, degeneration and separation of muscle fibers fibers are in agreement with the findings of earlier
were seen with the severity increasing with duration of workers in birds16. Atrophied spleen and depletion of
the experiment. In spleen,depletion of lymphocytes was lymphocytes noticed in imidacloprid treated groups may
noticed in white pulps. In ovary, congestion, disrupted be attributed to the immunotoxicity and immuno-
ovarian follicles and thecal cell depletion were observed, suppressive effect of imidacloprid, which lent support
while in uterus, loss of glandular epithelial cells with few to the earlier studies19,20. The changes of congestion and
cells having foamy cytoplasm and hyperplasia of thecal cell depletion in ovary, and loss of glandular
myometrium were observed.Brain sections revealed focal epithelial cells with cytoplasmic foaminessin uterus
areas of degeneration, mild glial cell proliferation and might be due to toxic effect of imidaclopridin the
satellitosis (Fig. 4). reproductive organs, and correlated with the observation
of earlier workers in birds and rats4,21-23. Brain revealed
In groups 3 and 5, all visceral organs showed
focal areas of degeneration, mild glial cell proliferation
insignificant changes except loss of islets in pancreas. In
and satellitosis, whicvh are in agreement with the
groups 4 and 6, kidneys revealed mild changes including
findings of an earlier study in rats17. Ultrastructural
Downloaded From IP - 117.211.167.37 on dated 20-Jun-2018
Ultrastructurally, in group 2, at 90 days, hepatocytes results correlated with the findings of a study on
revealed swollen nuclei, mild to moderate margination, imidacloprid-induced hepatotoxicity in male Albino
disruption and clumping of chromatin in the rats24. Relatively milder pathological changes or normal
nucleoplasm, thickened nuclear membrane, hyperplasia organs observed in group 4 and 6 could be attributed to
of rough endoplasmic reticulum and pleomorphic, the antioxidant property of vitamin C and Withania
condensed mitochondria. Kidneys showedswollen somnifera.
endoplasmic reticulum, ribosomes and mitochondria,
It is inferred from the study that supplementation of
with disrupted mitochondrial cristae (Fig. 5 and 6).
vitamin C @ 200 ppm in feed and Withania somnifera @
500 ppm in feed resulted in moderate protection to
DISCUSSION counteract the toxic effects of imidacloprid.
The results of gross abnormalities of swollen and
congested liver with rounded edges observed in group ACKNOWLEDGEMENTS
2in the present study, and histological changes of
The authors are thankful to Sri Venkateswara Veterinary
engorged sinusoids anddegenerative and necrotic
University for providing the facilities to carry out this work.
changes in the hepatocytes with Kupffer cell proliferation
were in agreement with the earlier findings observedin
birds9,15,16 and rats17,18. These changes might be due to REFERENCES
accumulation of imidacloprid metabolites in liver as it is 1. Al-saleh IA.1994. Pesticides: A review article. Journal of
the principal target organ for any detoxification Environmental Pathology, ToxicolOncol 13: 151-161.
mechanism. The vacuolation of hepatocytes could be due 2. Casale GP, Vennerstrom JL, Bavari S, Wang TI. 1993. Inhibition
to the retention of fluid inside the cell resulting in cloudy of interleukin 2 proliferation of mouse CTLL2 cells, by selected
carbamate and organophosphate insecticides and congeners
swelling probably due to reduction of energy supply of carbaryl. ImmunopharmacolImmunotoxicol 15: 199-215.
necessary for regulation of ion concentration of the cells15. 3. Soloway SB, Henry AC, Kollmeyer WD, Padgett WM, Powell
Liver of groups 4 and 6 showing congestion of portal JE, Roman CH, Tieman RA, Horn CA. 1978. Nitromethylene
vein and mild degenerative changes could be attributed heterocycles as insecticides. In Advances in Pesticide science.
to the free radical scavenging activity of vitamin C and Shankland DI, Hollingworth RM, Smyth T, Jr (Eds.) Pesticide
Venom Neurotoxicol, Plenum, New York, pp. 153-158.
Withania somnifera that might have resulted in mild
4. Brocksma A, Robberts E, Saba F. 1993. “Field trials with
regeneration and repair of damaged hepatocytes. confidor (imidacloprid) for the control of various insect species
The microscopic changes in kidney of imidacloprid on citrus in the Republic of South Africa: Pflanzenschutz-
Nachrichten. Bayer 46: 25- 32.
treated group 2 including interstitial hemorrhages,
5. Kutches AJ, Church DC,Duryee FL, 1970. Toxicological effects
tubular epithelial degeneration and shrunken glomeruli of pesticides on rumen functions in vitro. J Agri Food Chem 18:
with mesangeal cell proliferation may be due to the 430-433.
6
346 Babu et al.
6. Cerón JJ, Panizo CG,Montes A. 1995. Toxicological effects in 17. Bhardwaj S, Srivastava MK, Upasana, K, Srivastava LP., 2010.
rabbits induced by endosulfan, lindane, and methylparathion A 90 days oral toxicity of imidacloprid in female
representing agricultural by-products contamination.Bull ratsorphological, biochemical & histopathological evaluation.
Environ ContamToxicol 54:258-65. Food ChemToxicol 48: 1185-1190.
7. Matsumura F. 1985. Toxicology of insecticides. 2nd Ed., 18. Mohany M, Badr G, Refaat I, Garraud O, EI-feki M. 2012.
Plenum Press, New York, Pp. 267-270. Thymoquinone ameliorates immunological and histological
8. Abou-Donia MB, Goldstein LB, Bullman S, Tu T, Khan WA, changes induced by exposure to imidacloprid insecticide.
Dechkovskaia AM, Abdel-Rahman AA. 2008. Imidacloprid Toxicol Sci 37: 1-11.
induces neurobehavioral deficits and increases expression of 19. Balani T, Agarwal S, Thakar M. 2011. Haematological and
glial fibrillary acidic protein in the motor cortex and biochemical changes due to short term oral administration of
hippocampus in offspring rats following in utero exposure. J Imidacloprid – a neonecotinoid insecticide on immune system
ToxicolEnviron Health Part A71: 119-130. of white leghorn cockerels. J Vet Pharmacol Toxicol 7: 27-30.
9. Kammon AM, Brar RS, BangaHS,Sodhi S. 2010. Patho 20. Tripathi SM, Thaker AM, Joshi CG, Garg SP, Snakhla LN. 2007.
biochemical studies on hepatotoxicity and nephrotoxicity on Immunotoxicity Induced by Sub acute Acephate Exposure in
exposure to imidacloprid in layer chickens. VeterinarskiArhiv White Leghorn Cockerels. J IndianSocToxicol 3: 13-21.
80: 663-672. 21. PMRA (Pest Management Regulatory Agency). 2001.
10. Abd-El-Ghaney A. 2002.Study the effect of imidacloprid Imidacloprid. Regulatory Note. REG2001-11. Ottawa: Health
insecticide on some physiological parameters in Japanese Canada, Pest Management Regulatory Agency. Available at
quail. Thesis for M.Sc. Faculty of science Al-Azhar University. http://www.pmra-arla.gc.ca/english/pdf/reg/reg 2001-11-e.pdf
11. Feinstein RE. 2000. Post mortem procedures.In: Handbook of 22. Najafi G, Razi M, Hoshyar A, Shahmohamadloo S, Feyzi S.
Laboratory Animal Science.eds.SvendsenP, Hau J, pp.383-396. 2010. The effect of chronic exposure with imidacloprid
12. Luna GLHT. 1968. Manual of histological and special staining insecticide on fertility in mature male rats. Intl J Fert Ster 4: 9-
Downloaded From IP - 117.211.167.37 on dated 20-Jun-2018
13. BozzalaJJ,Russel LD. 1998. Electron Microscopy Principles Guvenc M, OzsahinAD, Kocaman N, Aslan A, Etem E. 2012.
www.IndianJournals.com
Techniques for Biologists. 2nd Edition. Jones and Barlett Assessment of imidacloprid toxicity on reproductive organ
Publishers, Sudbury, Massachusetts: pp. 19-45, 72-144. system of adult male rats. J Environ SciHlth, Part B47: 434-444.
14. Spurr AR. 1969. A low viscosity epoxy resin embedding 24. Soujanya S, Lakshman M, Kumar AA, Reddy AG. 2013.
medium for electron microscopy. J Ultrastr Res 26: 31-43. Evaluation of the protective role of vitamin C in imidacloprid-
15. Omiama SE. 2004. Protective effect of vitamin C and induced hepatotoxicity in male Albino rats. J Nat Sci Biol Med
glutathione against the histopathological changes induced by 4: 63-67.
imidacloprid in the liver and testis of Japanese quail. Egyptian
JHosp Med 16: 39-54.
16. Subha Ganguly. 2013. Long term exposure to chemicals,
insecticides and heavy metals causing toxicity: A Review. IntJ
Pharma Res Bio-Sci 2: 333-342