ANJANEYULU, Y(MAJOR)Srilatha, ChBHARATH, B.K2018-10-252018-10-252008-11http://krishikosh.egranth.ac.in/handle/1/5810082078THESESABSTRACT : India has highest animal population in the world and animal husbandry occupies a prominent position in agricultural sector.It has been one of the subsidiary occupation for rural people offering socioeconomic stability and is one of the major contributors to the national economy. Indiscriminate use of pesticides to control pests in various agricultural and animal husbandry practices and vector control programmes of public health concern may lead to health hazards due to their presence in food of human beings and livestock. Endosulfan is a chlorinated hydrocarbon pesticide under the cyclodiene subgroup. It emerged as a leading chemical used against a broad spectrum of insects and mites in agriculture and allied sector. It acts as contact and stomach poison and has slight fumigant action. It is used in vegetables, fruits, paddy, cotton, cashew, tea, coffee, tobacco, timber crops and also as a wood preservative. Exposure to endosulfan can induce a number of effects including neuro, hepato and nephrotoxicity, haematological and biochemical effects, altering the immune system and damage the reproductive organs. Most of the earlier studies are restricted to acute effects of pesticide in experimental animals. However the situation in practice is not toxicity resulting from a single or a few large doses of a given pesticide, but due to oral intake of very small quantities over a reasonably long period of time. Studies related to endosulfan toxicity have been carried out in different animals. There is paucity of detailed information regarding endosulfan induced chronic toxicity and also ameliorative effect of O S against endosulfan, hence a systematic work was undertaken to study the toxic effects of low levels of endosulfan and ameliorative effect of Ocimum sanctum against endosulfan induced damages and immuno suppression with the objectives to study the effect of endosulfan on haematobiochemical, gross and histopathological changes in different organs and to study the ultrastructural changes in liver and RBCs and ameliorative effect of Ocimum sanctum against endosulfan induced damages. The present experiment was designed to make a systematic study of experimentally induced endosulfan toxicity in male rats. Technical grade endosulfan was given at 6, 3 and 1.5 mg / Kg b.wt to groups II, III and IV by mixing in ground nut oil for 6 weeks, to the groups V, VI and VII in addition to endosulfan, Ocimum sanctum was given @200 mg / Kg b.wt daily per orally for the same duration. Group I and VIII served as oil and Ocimum sanctum control. Specific clinical signs were not observed, except few rats of the Group II and Group V. Showed almost similar clinical signs in general during the experimental period which included dullness, depression, diarrhoea, intermittent anorexia and hyper excitability. In the present investigation no mortality was observed in any of the treated group. Significant (P<0.05) reduction in the weight gain was observed in toxin treated groups when compared to control groups and these values are nearer to normal in lower doses of endosulfan with OS ameliorated groups. Significant (P<0.05) reduction in the PCV, Hb, TEC, TLC, percent lymphocytes count and significant increase in the percent neutrophil count were observed in all the endosulfan treated groups. These counts were significantly improved in the OS ameliorated groups with lower dose of endosulfan (groups VI and VII). Significant (P<0.05) increase in serum glucose, creatinine, cholesterol, ALT, AST, and significant (P<0.05) decrease in the TSP levels in endosulfan treated groups when compared to control groups in a dose dependant manner. And these levels were in between the above groups in the OS ameliorated groups. Significant (P<0.05) decrease in the serum T3 and T4 levels were noticed in all toxin treated groups when compared to the control groups in a dose dependent manner, and there was no change in TSH levels in any of the groups. In OS treated groups significant improvement was observed compared to the toxin treated groups in T3 and T4 levels. Significantly (P<0.05) decreased testicular wet weights, sperm counts were observed in all the experimental group when compare to the Oil control group. Significant decrease in the HA titer (log) and DNCB contact sensitivity score in groups II, III and V when compare to the control groups and these values are in between the above groups in IV, VI and VII groups. In the present study gross lesions were prominently observed in the liver followed by kidney, lung, spleen, brain and heart. Liver revealed moderate enlargement during the 2nd week. Severe enlargement of the liver with rounded borders and paleness of the liver was observed after 4th and 6th week in all endosulfan treated groups in a dose dependant manner and changes are less severe in OS ameliorated groups. Kidneys revealed enlargement with moderate congestion in all the endosulfan treated groups throughout the experimental period. Lungs of the rats from the endosulfan treated groups revealed moderate to severe congestion throughout the experimental period. With less severity in OS treated groups. Severe enlargement of spleen was observed during the 2nd week of the experiment but after 4th and 6th week observed the reduction in the size of the spleen in endosulfan treated groups with less severity in the OS groups. Brain showed mild to moderate congestion in all the experimental animals throughout the experiment. Mild enlargement of the heart was observed in all the endosulfan treated groups throughout the experimental period. Histopathologically the liver of rats fed with endosulfan for 2 weeks revealed congestion, degenerative changes in hepatic cells with cellular swelling, mild fatty changes, focal MNC aggregation, mild perivascular and periportal MNC infiltration, sinusoidal dilatation with haemorrhages and focal loss of hepatocytes with MNC infiltration and mild bile duct proliferation. In addition to above changes mild to moderate fatty changes, few areas of hyper chromatic nuclei, bi nucleated hepatocytes, loss of hepatocytes with dilated sinusoids and micro granulomatous lesions, MNC infiltration around the blood vessels with fibroblast proliferation by the end of 4th week were noticed. More number of bi nucleated hepatocytes, individualization of hepatocytes, bile duct proliferation with hyperplasia of epithelium, karyomegaly and focal areas of centri lobular necrosis were observed very conspicuously by the end of 6th week in a dose dependant manner and these lesions were mild in the OS ameliorated groups with lower doses of endosulfan (group VI and VII). Microscopic examination of kidneys treated with endosulfan revealed dose dependent congestion of blood vessels and glomeruli, haemorrhages in between the tubules, mild fibroblast proliferation, degenerative changes in the tubular epithelium and mild infiltration of MNCs by the end of 2nd week. In addition desquamation of epithelium leaving only basement membranes, severe haemorrhages, loss of tubules leaving cystic spaces, perivascular edema with fibroblast proliferation, karyomegaly, focal aggregation of MNCs and atrophy of glomerular tuft were observed by the end of 4th week. In addition hyaline casts in the lumen of tubules, some tubules with hyperplasia of tubular epithelium, in some areas total loss of tubules with haemorrhages and lytic changes around the glomeruli and capillaries and cystic glomeruli were observed by the end of 6th week. All these changes were dose and time dependent and these changes are less severe in OS treated groups in lower dose groups. Congestion, thickened blood vessels, mild to moderate edema, perivascular and peribronchiolar edema, haemorrhages, emphysematous changes and inflammatory cell infiltration were observed by the end of 2nd week. In addition to above hyperplasia of bronchiolar epithelium, hyalinised blood vessels, peribronchiolar fibrous tissue proliferation, thickened interstitial space due to infiltration of MNCs, RBCs and fibroblast by the end of 4th week were noticed. Hyper trophy and hyperplasia of bronchiolar lymphoid follicles, MNCs infiltration around the blood vessels, peri bronchial regions aggregation of the macrophages, plasma cells and other MNCs and haemorrhages around the blood vessels were observed by the end of 6th week all these changes are dose and time dependent. In OS ameliorated groups these changes were mild compare to the corresponding toxin treated groups. Cerebrum of rats treated with 6mg/kg b.wt endosulfan showed congested blood vessels, focal minute haemorrhages in the cerebrum, by the end of 2nd week. In addition proliferation of capillaries, mild to moderate gliosis, shrinkage of neurons, central chromatolysis, neuronophagia, micro nodular lesions in the cerebrum, sub meningeal haemorrhages in cerebrum and cerebellum region, prominent chromatolysis, gliosis, neuronophagia, demyelinating changes, vacuolation, shrinkage of neurons, focal loss of grey matter with aggregation of glial cells by the end of 4th week. Sub meningeal accumulation of MNCs, sever gliosis, vacuolation, thickening of meninges with fibroblasts, in certain areas and Purkinjee cell hyperplasia were observed, Central chromatolysis of neurons was more conspicuous in rats sacrificed by the end of 6th week. Cerebellum of rats congested blood vessels, focal minute haemorrhages focal loss of purkinjee cells, sub meningeal haemorrhages, grouping of purkinjee cells and central chromatolysis at the end of 4th and 6th week. Testis showed congestion, interstitial edema, separation of seminiferous tubules and degenerative changes in the seminiferous tubules by the end of 2nd week. Perivascular edema, loss of tubular epithelium leaving only basement membranes with sertoli cells and degeneration with desquamated cell debris within the lumen of tubules, MNCs and plasma cell infiltration in the interstitial spaces, hyalinization of blood vessels and few seminiferous tubules were observed by the end of 4th week. Complete separation of germinal epithelium from basement membrane, severe degenerative and lytic changes were observed by the end of 6th week. All these changes were dose and time dependent and these changes were severe in OS treated groups. Spleen of group II and III rats revealed congestion, mild lymphoid depletion by the end of 2nd week, engorgement of red pulp with erythrocytes, severe congestion of vessels with moderate to severe depletion of lymphocytes and reduced number of lymphoid follicles at 4th week to 6th week was observed. In other groups mild lymphoid depletion with congestion of vessels was observed and in spleen of group VII rats only mild depletion was observed when compare to the control groups. In group II and V rats revealed changes like congestion of blood vessels, mild to moderate increase in number of goblet cells, edema, desquamation of epithelium and areas necrosis with MNCs infiltration by the end of 6th week. Group III and VI rats revealed changes like mild increase in number of goblet cells and mild mononuclear cell infiltration. These changes were mild in OS ameliorated groups compared to toxin treated groups. Pancreas of all the endosulfan treated rats showed dose dependant congestion of blood vessels, haemorrhages and mild degenerative changes both in acini and Islets of Langerhans by the end of 2nd week. In addition hyalinised blood vessels, infiltration of MNCs within and between the follicles, severe vacuolar degeneration of both endocrine and exocrine part, atrophy and necrosis of Islets, epithelial hyperplasia of intercalated ducts, fibroblast proliferation around the blood vessels, ducts and in between the follicles, atrophy of the acini were observed by the end of 6th week. Similar lesions were observed in the group V as that of group II but these changes was less severe in group VI and VII. Adrenals cortex of endosulfan treated groups showed dose dependent vacuolar degeneration, telengectasis, swollen foamy cytoplasm, few bi nucleated cells, sub capsular and medullary haemorrhages. Very mild changes were observed in OS ameliorated groups. Lymph nodes of all the experimental groups showed dose and time dependent congestion, haemorrhages, edema and cortical depletion. And the depletion was not so severe compare to the endosulfan treated alone in the lower dose with O S ameliorated groups. Scanning electron microscopy of RBCs treated with endosulfan revealed dose dependent changes like loss of normal cellular outlines with membrane fusions, crenations, surface pits, loss of biconcavity and threading of membranes by the end of 4th and 6th week. Where as in OS treated groups these changes were restored towards normal. Transmission electron microscopy liver treated with endosulfan revealed swollen nucleus, clumping of outer and inner nuclear membrane, scanty chromatin material, vacuolation of cytoplasm, reduction in the number of mitochondria and endoplasmic reticulum, fragmentation and condensation of chromatin material by the end of 4th and 6th week on dose dependant manner compare to the control. In higher dose levels of toxin in the amelioration dense presence of mitochondria and endoplasmic reticulum were noticed.ennullThesis