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2010 - 20174
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Subject: Veterinary Pathology × End date: 2018 × Start date: 2010 × Thesis Type: Ph.D ×
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    ThesisItemOpen Access
    MOLECULAR AND PATHOLOGICAL STUDIES OF INDUCED HYPERLIPIDEMIA AND ATHEROSCLEROSIS IN RATS AND ITS AMELIORATION WITH FLAXSEEDS (LINUM USITATISSIUMUM) AND GREEN TEA (CAMELLIA SINENSIS)
    (SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2017-05) SRINIVASA NAIK, H; SRILATHA, Ch(MAJOR); SUJATHA, K; SREEDEVI, B; PRASAD, T.N.V.K.V
      ABSTRACT : Hyerlipidemia is the disorder of lipid metabolism, characterized by elevated serum total cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL-C) and very low density lipoproteins cholesterol (VLDL-C) and decreased high density lipoprotein cholesterol (HDL-C). Atherosclerosis is diverse disease with heterogenous mechanisms of progression and is an oxidative, chronic inflammatory and thrombotic disease referring to fatty deposits on the inner lining of the large to medium sized elastic and muscular arteries and is precipitated by elevated level of low density lipoprotein cholesterol (LDL-C) in the blood. Atherosclerosis associated coronary heart disease is the second largest cause of mortality worldwide in humans. Current reports suggest that by the year 2020, India will have the largest cardio vascular disease (CVD) burden in the world. Realizing the importance of ethno medical practices, flaxseed and green tea supplementation along with atherogenic diet has been taken to combat the hyperlipidemia associated atherosclerosis. Flaxseed (Linumusitatissiumum) has been identified as a significant alternative source of omega 3 fatty acids and alpha linoleic acid (ALA). Therapeutic activities of flaxseed is well proven, one of them is anti-atherosclerotic due to its lipid lowering activity which is mainly attributed to its Secoisoarciresinol diglucoside (SDG). Green tea (Camellia sinensis) polyphenols known to have various medical health beneficial effects like anti-inflammatory, anti-oxidative, anti-arthritic, anti angiogenic, anti-metastatic, anti-cancer, anti-obesity, anti hyperlipidemic, anti-atherosclerotic, neuroprotective, anti-dental caries and antimicrobial (bacterial, viral & fungal) properties in human, animal and in vitro studies. So the present study was designed with a view to study molecular aspects of hyperlipidemia associated atherosclerosis in male Wistar albino rats and an attempt was made to identify various molecular preventive effects of flax seeds (Linumusitatissiumum) and green tea (Cameliasinensis). The present study was carried out by procuring 72 wistar albino rats, that were randomly divided into six groups consisting of 12 rats in each group. Hyperlipidemia associated atherosclerosis was induced by atherogenic diet consisting of 1% cholesterol and 15% saturated oil added to the 1000g of standard rat diet and given to group II rats. Flaxseed @ 7.5g/kg/day and green tea @ 100mg/kg/day gavaged orally to group V and VI rats for 90 days along with atherogenic diet. Group I kept as control and given standard rat diet, group III as flaxseed control and given 7.5g/kg/day of flaxseed along with standard rat diet, whereas group IV kept as green tea control and given 100mg/kg/day/Po along with standard rat diet for 90 days. Six rats from each group were sacrificed 45 days apart each. Atherogenic diet fed group (Group II) rats clinically showed obesity with significant increase (p<0.05) in the body weight. Rats were sluggish with poor hair coat. Green tea supplementation (Group VI) significantly reduced the obesity and body weight compared to moderate reduction of obesity and body weight in flaxseed ameliorated group (Group V). Whereas green tea control group (Group IV) rats were apparently healthy, slim with shiny hair coat. Total leukocyte count (TLC), total cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL-C), very low density lipoprotein cholesterol (VLDL-C) were significantly (p<0.05) higher, whereas high density lipoprotein cholesterol (HDL-C) was significantly reduced in group II. Oxidative damage indicators like catalase, SOD, GPx, reduced glutathione and glutathione S transferase were significantly (p < 0.05) reduced, whereas lipid peroxidation products of TBARS level was non-significantly (p < 0.05) increased in liver and heart tissues of atherogenic diet fed group (Group II). Flaxseeds supplementation (Group V) reduced the hyperlipidemia certain extent caused by atherogenic diet by reducing the modest level of TC, TG, LDL-C and mild elevation of HDL-C. Flaxseeds non-significantly (p < 0.05) increased the cellular anti-oxidant enzymes and modestly reduced the level of TBARS. Whereas green tea (Group VI) supplementation effectively controlled the hyperlipidemia by reducing all the serum total cholesterol, triglycerides, low density lipoprotein cholesterol and very low density lipoprotein cholesterol and enhanced the good high density cholesterol and significantly (p < 0.05) enhanced all the tissue anti-oxidant enzymes and modestly reduced the TBARS levels. Microscopically hepatic steatosis was prominent in atherogenic diet fed group (Group II). Complete absence of fatty change was evident in green tea treated group (Group VI) compared to flaxseed group (Group V). Histopathologically, endothelial degeneration, sub intimal fat cells, foam cells with attached microthrombi was observed in majority of the cases in group II rats aorta fed on atherogenic diet. All these changes were also evidenced by ultrastructural studies. Mild endothelial degeneration was evident in flaxseed ameliorated group. Majority of the changes of initiated atherosclerosis were minimized by green tea supplementation. No such lesions were seen in control, flaxseed and green tea alone fed control groups (Group I, III & IV) throughout the study period. Mild endothelial cell proliferation with formation of vascular channels in between cardiac muscle fibers, liver and in kidney was observed throughout the study period of flaxseed fed group (Group III) rats. VEGF positivity was observed in endothelial cells, thrombus and structurally modified stromal cells of initiated atherosclerotic lesion. Initiated atheromatous plaque showed high level of positivity with Bax compared with other parts of blood vessel. On the other hand, selective moderate to severe positivity with CD 31, CD44, E cadherin and Beta catenin was observed in the initiated atherosclerotic lesion characterized by endothelial degeneration, sub endothelial lipid laden macrophages, and endothelial adhered thrombus and erythrocytes. Various levels of positivity with above markers was also observed in flaxseed and green tea ameliorated groups based on the initiated atherosclerotic lesion in these groups. Present study revealed positive expression of proinflammatory cytokine TNF alpha by RT-PCR (Real time PCR) in atherogenic diet fed group aorta (Group II) compared to standard diet fed rats (Group I). Flaxseed and green tea ameliorated groups (Group V and VI) showed significant reduction in the expression of TNF alpha in contrary to atherogenic diet fed group (Group II). IL-18 expression was not noticed in any groups of the present study. In conclusion atherogenic diet of present study established the hyperlipidemia and initiated the atherosclerosis in the aorta evidenced by histopathology and molecular expression of PECAM-I (CD 31), CD44, E cadherin and Beta catenin and tissue TNF alpha. Flaxseeds supplementation reduced the hyperlipidemia associated atherosclerosis to certain extent, but not completely ameliorated the changes. On the other hand, green tea supplementation effectively controlled the hyperlipidemia associated atherosclerosis and enhanced all the tissue anti-oxidant enzymes.
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    ThesisItemOpen Access
    STUDY ON TOXIC EFFECTS OF IMIDACLOPRID AND ITS AMELIORATION IN LAYER BIRDS
    (SRI VENKATESWARA VETERINARY UNIVERSITY , TIRUPATI – 517502. (A.P.) INDIA, 2014-05) SASIDHAR BABU, N; ANAND KUMAR, A (Major); SRILATHA, Ch; SREENIVASULU, D; MOULI KRISHNA, K
    ABSTRACT : The present study on “Toxic effects of imidacloprid and its amelioration in layer birds” at definite time periods in different groups for 90 days was designed to elucidate the haematological, biochemical, immunological, oxidative stress, histopathological as well as ultra-structural alterations induced by imidacloprid and also to evaluate the protective effect of vitamin C and Withania somnifera on imidacloprid induced toxicity. The study was carried out on 48 layer birds of above 2 months age that were divided into six groups consisting of 8 in each. The experiment was carried out according to the guidelines and prior approval of Institutional Animal Ethics Committee. All the birds were provided with standard diet and deionized water ad libitum, throughout the experimental period of 90 days. All the experimental birds were placed in cages in the Department of Poultry Science and allowed to acclimatize for about week days and were observed thrice daily for clinical signs and mortality if any, during the entire period of study. The experimental design is as follows: Group 1 served as control, group 2 was treated with imidacloprid @ 200 ppm in feed, group 3 was treated with vitamin C @ 200 ppm in feed, group 4 was treated with both imidacloprid@200ppm and vitamin C 200ppm, group 5 was treated with Withania somnifera @ 500 ppm in feed and group 6 was treated with imidacloprid @ 200 ppm in feed + Vitamin C @ 200 ppm in feed + Withania somnifera @ 500 ppm in feed. Clinical signs exhibited by the birds during the course of experiment were, the birds in groups 1, 3, and 6 were apparently healthy and the birds in group 2 were apparently healthy up to the end of 30 days, but later they started showing clinical symptoms like reduced feed intake, dullness, diarrhoea, decreased size of eggs and reduced production, with increasing severity as per the advancement in age. Reduction in body weights was observed in group 2 in comparison to group 1 and in groups 4 and 6 weights increased significantly (P < 0.05) in comparison to group 2. Haematological studies revealed insignificant (P < 0.05) decrease of TEC, Hb concentration and PCV, TLC and DLC were significantly (P < 0.05) decreased in birds of group 2 in comparison to groups 1, 3 and 5. In groups 4 and 6, insignificant (P < 0.05) decrease in all the above parameters was noted in comparison to group 2. The biochemical assays showed a significant (P < 0.05) reduction of glucose, total protein and albumin in group 2 in comparison to groups 1, 3, 5 & 6. In group 4, no significant difference was observed but in group 6 significant difference was observed when compared to group 2. In groups 2, 4 and 6, the serum creatinine, BUN levels, AST and ALT activity were significantly (P < 0.05) increased when compared to groups 1, 3 and 5. The tissue biochemical profile revealed a significant (P < 0.05) reduction in GSH concentration and increase in TBARS levels in liver of group 2. Group 3 value was insignificant from control and group 4 showed a significant (P < 0.05) increase in comparison to group 2. Grossly, group 2 birds on day 60 showed congested and enlarged livers with rounded boarders and atrophied spleens and the severity of the lesions were slightly higher on day 90. Histopathologically, in group 2, at 30 days- mild fatty change, mild degenerative and necrotic changes in the hepatic lobules were observed. At 60 days- all the above mentioned lesions were moderate and focal areas of franc necrosis were noted. At 90 days – prominent lesions like moderate to severe fatty change, central vein congestion, engorged sinusoids, severe degenerative and necrotic changes and focal aggregation of lymphoid cells was recorded. In group 4, at 60 days- mild degenerative changes and at 90 days- mild to moderate degeneration in the hepatic lobules was noted. In group 6, at 60 days and 90 days– mild degenerative changes in the hepatic lobules were noted. In group 2, kidney at 30 days- inter-tubular hemorrhages was observed. At 60 days- congestion, inter-tubular hemorrhages, moderate tubular degeneration, atrophied glomeruli and focal necrotic areas was noted. At 90 days – prominent / severe lesions like tubular degeneration, atrophied glomeruli, mesangeal cell proliferation and vacuolation was recorded. In group 4, at 30 days - degenerative changes in tubules are observed. At 60 days- mild to moderate changes, inter-tubular congestion and hemorrhages and at 90 days- moderate lesions were noted. In group 6, at 90 days– mild degenerative changes in the tubules was noted. In group 2, hearts, at 30 days- mild degeneration and separation of muscle bundles was observed. At 60 days- moderate degeneration and separation of muscle bundles was noted. At 90 days- moderate to severe lesions like endocardial hemorrhages, degeneration and separation of muscle bundles was recorded. In groups 4 and 6 at 60 days- mild degenerative changes and at 90 days- moderate degenerative changes in cardiac muscle bundles was noted. In gizzard and proventriculus, no lesions of pathological significance were observed in any of the groups at any point of time. Ruptured villi in intestines, loss of islets in pancreas, depletion of lymphocytes in spleen, mild to moderate congestion and disrupted ovarian follicles, mild to moderate loss of glandular epithelial cells in uterus with few epithelial cells showing foamy cytoplasm and hyperplasia of myometrium and in brain focal areas of degeneration, mild glial cell proliferation and satellitosis in group 2 was recorded at 90 days. The lesions were improved in groups 4 and 6. Ultrastructurally, in group 2, at 90 days ultra-thin sections of liver revealed swollen nuclei, mild to moderate margination, disruption and clumping of chromatin in nucleoplasm, collapsed and thickened nuclear membrane, hyperplasia of rough endoplasmic reticulum, peri-nuclear and intercellular cell junctions fat depots, degenerated hepatocytes, thick, vesicular cytoplasm and pleomorphic, condensed mitochondria. Sections of kidney showed loss of endothelial and epithelial cells, vesicular nuclei, degeneration of tubular epithelium with loose inter cellular junctions, swollen, disrupted nuclei, margination of chromatin material (apoptosis), vacuoles in the cytoplasm, pleomorphic, pyknotic nuclei, disturbed shape and size of mitochondria. The studies on imidacloprid toxicity @ 200 ppm for 90 days in birds resulted majorly in nephrotoxicity, hepatotoxicity, toxicity on reproductive organs, followed by mild to moderate cardiac and neuro toxicity. Moderate immunosuppression was exhibited. Altered body weights, haematological, biochemical, histological and ultrastructural parameters were recorded which might be due to imidacloprid- induced oxidative stress that resulted in depletion of GSH. The alterations in biochemical parameters were supported by histological and ultrastructural changes in kidney and liver. Supplementation of vitamin C @ 200 ppm in feed and Withania somnifera @ 500 ppm in feed resulted in moderate protection by repair and regeneration of damaged tissues to counteract the toxic effects of imidacloprid.
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    ThesisItemOpen Access
    STUDIES ON PATHOLOGY OF BISPHENOL – A TOXICITY IN RATS WITH SPECIAL REFERENCE TO REPRODUCTIVE SYSTEM
    (2011-08) AMARAVATHI, P; SRILATHA, Ch (Major); RAMA DEVI, V; REENIVASULU, D
    ABSTRACT : Bisphenol A (BPA) is one of the common environmental endocrine disruptors with estrogenic properties and is the building block of carbonate plastic and a component of resin coatings. Wide spread use of BPA in consumer products has led to great public concern since adverse effects of BPA on human and animal reproduction are suspected due to its estrogenic activity. BPA has high affinity to estrogen related receptor (ERR -) which may be related to its ability to function as endocrine disruptor. Exposure to BPA can induce a number of effects including neuro and hepato toxicity, hematobiochemical effect, endocrinal disturbances, genotoxicity, reproductive toxicity and alters the immune system. The present experiment was designed to make a systematic study of experimentally induced BPA toxicity in both male and female Wistar albino rats at 500 and 250 mg /kg b.wt. to groups II , V and III , VI respectively by mixing in sunflower oil for 12 weeks, with the objectives of finding out the effect of BPA toxicity on hemato- biochemical, oxidative damage, hormonal assay, reproductive toxicity, immunological, genotoxicity, gross and histopathological and ultra structural changes.. Specific clinical signs were not observed, except reddish discoloration of hair was noticed around nose and on back region of BPA treated female rats. Hematologically significant (P<0.05) reduction was recorded in the Hb, PCV and TLC values in toxin fed groups. Blood glucose, and serum total protein values were decreased significantly (P<0.05) in BPA treated rats. Oxidative damage indicators like SOD, catalase and GPx levels were decreased in liver and kidney of all the BPA treated groups. Hormonal assay revealed significant (P<0.05) increase in T3, T4 and E2 levels in BPA fed groups. Immunosupression was indicated by significant (P<0.05) decrease in HA titer (log) and DNCB contact sensitivity score. DNA damage was evident as significant (P<0.05) increase in micronuclei in PCE cells of BPA treated groups when compared to control. In the present study grossly moderate enlargement of liver with rounded borders with paleness and reduction in size of testis were observed in all BPA treated groups in a dose dependent manner. Enlargement of spleen was observed during 2-4 weeks of the experiment but by the end of experiment, reduction in the size of the spleen was prominent in BPA treated groups. Enlargement of heart was noticed in BPA treated male rats from 6th week onwards and it was more conspicuous by the end of 12th week and this lesion was not more conspicuous in female rats. Histopathologically, the liver revealed binucleated cells, hyper chromatic nuclei, karyomegaly, focal loss of hepatocytes with moderate MNC infiltration, moderate to severe vesicular fatty change, extensive bile duct proliferation with dysplasia and proliferation of endothelial cells in BPA treated groups in dose dependent manner. Kidney sections of BPA treated rats revealed dose dependent congestion of glomeruli, tubular degeneration, glomerular edema and mesangial cell proliferation. Submeningeal and cerebral hemorrhages and spongiosis, swollen neurons with severe degenerative changes, extensive glial cell proliferation, severe demyelinating changes, microgranuloma formation with proliferation of capillaries and endothelial cells were more prominent in cerebral cortex of majority of BPA treated groups in a dose dependent manner. Cerebellum of rats revealed shrinkage and focal loss of Purkinje cells, rounding and tapering of Purkinje cells, spongiosis in molecular layer in majority of animals at the end of 10th to 12th weeks in BPA treated groups. Microscopically, lungs revealed congestion, fatty infiltration, peribronchial lymphoid aggregates and infiltration of eosinophils, vacuolated alveolar epithelial cells, infiltration of alveolar macrophages, hyperplasia of bronchiolar epithelium and focal alveolar epithelial cell proliferation with papillary projections and desquamated bronchial epithelial cells were found as constant lesion by the end of 12th week in a dose dependent manner. Testis showed interstitial edema, shrinkage of tubules, basement membrane with spermatogonial cells, desquamated seminiferous tubular germ cells, vacuolation in sertoli cells and presence of multinucleated cells in seminiferous tubules were conspicuous by the end of 12 weeks. Hyperplasia of seminal vesicles with papillary projections and prostate carcinoma were noticed in BPA treated rats. Degenerated follicles, granulosa cell tumor, androblastoma changes were noticed in ovaries of BPA treated rats. Sections of uterus revealed degenerative changes in endometrial glands, infiltration of mono nuclear cells, eosinophils and plasma cells, adenomyosis, dysplasia of endometrial glands and cystic glands by the end of 12th week in majority of BPA treated rats in dose dependent manner. In sections of mammary glands degenerated acini with desquamated epithelial cells in lumen and proliferated acinar epithelial cells were observed in BPA treated rats. Pancreas revealed degenerated Islets of Langerhans, thickened and hyalinized blood vessels, hyperplasia of ductular epithelium, focal MNC infiltration, necrosis and atrophy of islets, vacuolar degeneration in acinar epithelium and interlobular hemorrhages were more conspicuous in majority of animals in BPA treated groups when compared to control group by the end of 12th week. Thyroid of BPA treated rats revealed dose dependent changes like hemorrhages, severe desquamation of acinar epithelial cells, complete absence of colloid in acini throughout experimental period and thyroid adenocarcinoma was observed by the end of 12th week. Degenerative changes in adrenal gland and depletion of lymphocytes in spleen, lymph node and thymus were noticed in all BPA treated rats in dose dependent manner. Skin of BPA treated groups revealed mild MNC infiltration in dermis, thinning of epidermis, cystic hair follicles and hyperplastic hair follicular epithelium with infiltration of eosinophils in dose dependent manner. Histochemically more intense alkaline phosphatase reaction was noticed in hepatocytes around central vein, in tubular epithelium and basement membrane of proximal convoluted tubules of kidney and spermatogonia of seminiferous tubules of BPA fed groups in dose dependent manner. Immunohistochemically increased expression of VEGF was observed in hepatocytes around central vein, in the lining epithelium of oviduct and endometrial glands of BPA treated rats in dose dependent manner. Increased expression of Bcl2 was observed in endometrial glands of BPA treated rats. Ultra structurally, hepatocytes of BPA treated groups revealed decreased mitochondria with degeneration, fragmented endoplasmic reticulum, and clumping of nuclear chromatin. Where as kidneys revealed loss of brush border, flattened nucleus, degenerated mitochondria and chemical deposition. Granular mitochondria because of degeneration of cristae, degeneration of ER, vacuolation in cytoplasm and fragmented chromatin in astrocytes and numerous vacuoles in cytoplasm with disrupted nuclear membrane and condensation of chromatin in microglial cells were noticed in brain of BPA treated rats. Electron microscopic examination of sertoli cells revealed decreased number of cell organelles and few mitochondria in cytoplasm and disrupted nuclear membrane in all BPA treated male rats in dose dependent manner.
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    ThesisItemOpen Access
    PATHOMORPHOLOGICAL STUDIES ON LEAD POISONING IN WISTER ALBINO RATS WITH SPECIAL REFERENCE TO REPRODUCTIVE TOXICITY AND ITS AMELIORATION
    (Sri Venkateswara Veterinary University, TIRUPATI – 517 502,A.P, 2010-04) SUJATHA, K; SRILATHA, Ch (Major); ANJANEYULU, Y; CHANDRA SEKHARA RAO, T.S; SRINIVASULU, D
    ABSTRACT: Lead is an abundant, ubiquitous, dangerous and important toxic environmental contaminant and is of global concern due to its significant role in modern industry. Lead is the most important poison to the farm animals and the significant pollution is likely to occur from lead mining, painted and metallic lead in storage batteries, licking paints / puttyans from rubbish dumps. Exposure to lead can induce a number of effects including neuro, hepato and nephrotoxicity, hematobiochemical effect, endocrinal disturbances, genotoxicity, reproductive toxicity and alters the immune system. The present experiment was designed to make a systematic study of experimentally induced lead poisoning and its amelioration with Ocimum sanctum in male Wister albino rats at 60 mg/kg bwt / 3 days in a week and 30 mg /kg bwt / 3 days in a week to groups II and III respectively by mixing in distilled water for 12 weeks. To the groups IV and V in addition to lead acetate as above mentioned dose, Ocimum sanctum was given at 400 mg / kg bwt/3 days a week / orally for the same duration, with the objectives of finding out of the effect of lead poisoning on hematobiochemical, oxidative damage, hormonal assay, reproductive toxicity, immunological, genotoxicity, gross and histopathological, ultra structural changes, residual estimation and ameliorative effect of Ocimum sanctum against lead induced damages. In addition epidemiological study on lead content in the blood and milk samples of cattle in five different factory areas as well as tissue samples of cattle and sheep form different urban areas was carried out. Specific clinical signs were not observed, except few rats of group II, III, IV and V that showed almost similar signs in general during the experimental period as dose dependent manner which included reduced feed intake, anxiety, loss of muscle coordination, tremor, dizziness etc. In the present investigation no mortality was observed in any of the treated group. Significant (P<0.05) reduction was recorded in the TEC, PCV, Hb and percent lymphocyte count. Significant increase in the percent neutrophil count and TLC were observed in all the lead treated groups. These counts were significantly improved in the OS ameliorated groups with lower dose of lead acetate (group V). Significant (P<0.05) decrease was noticed in the CAT, SOD, GPx activity in liver, kidney and testis of lead treated groups when compared to control groups. Non significant improvement in these values was observed in ameliorated groups (V & V) in a dose dependent manner. A significant (P<0.05) decrease was noticed in T3, T4 and TSH levels, acetyl cholinesterase activity in brain and significant increase in serum testosterone and LDH levels in lead treated groups (Group II & III).The level of T3 and TSH were significantly improved in the OS ameliorated groups with lower dose of lead acetate (Group V), while the remaining values were non significantly improved in OS ameliorated groups. Significantly (P<0.05) decrease in testicular weight, sperm count and significant increase in abnormal sperm count were observed in lead treated groups when compared to control group. Significant (P<0.05) decrease was noticed in HA titer (log) and DNCB contact sensitivity score in groups II and III when compared to control group. In OS ameliorated groups (IV & V) a significant improvement was noticed in these values. Significant (P<0.05) increase was seen in micronuclei in PCE cells of lead treated groups (Group II & III) when compared to control. In OS ameliorated groups (IV & V) a significant decrease was noticed in these values. Histopathlogically, the liver of group II rats revealed sinusoidal dilatation dilated central vein and focal areas of necrosis, hyper chromatic nuclei, binucleated cells, karyomegaly. Focal loss of hepatocytes with moderate MNC infiltration, moderate to severe vesicular fatty change, extensive bile duct proliferation and fibroblast proliferation around periportal area. Microgranulomas were observed very conspicuously in lead treated groups in a dose dependent manner and these lesions were mild in the OS ameliorated group (Group IV) with higher dose of lead after 6th week of lead feeding. The changes were gradually reduced and by the end of 10th week, it regains its normal appearance. Where as in Group V (OS ameliorated group of lower dose) these changes were less intensive and by the end of 8 to 12 weeks, liver comes to near normal. Kidney sections of lead treated rats revealed dose dependent congestion of glomeruli, mild to moderate hemorrhages, glomerular vacuolation, atrophy of glomeruli, desquamated tubular epithelial cells, pockets of hemorrhages in cortex and severe mononuclear cell infiltration were observed by the end of 12th week. All the changes were dose and duration dependent and these changes were less severe in OS treated groups. Extensive submeningeal and cerebral hemorrhages and spongiosis, swollen neurons with severe degenerative changes, glial cell proliferation, severe demyelinating changes, gliosis, infiltration of MNC, perivascular infiltration of MNC and necrotic nodules with proliferation of capillaries were more prominent in cerebral cortex of majority of lead treated groups (groups II and III) in a dose dependent manner. Cerebellum of rats showed spongiosis, shrinkage and focal loss of Purkinje cells, rounding and tapering of Purkinje cells, demyelination and vacuolatory changes in majority of animals at the end of 10th to 12th weeks in lead treated groups. These changes were less severe in OS ameliorated groups of higher dose and lower dose (IV & V). Microscopically, Group II and III lungs revealed congestion, edema, hyalinised blood vessels and bronchiole, intense emphysema, widening of interstitial spaces with MNC, edema and RBC, swollen and vacuolated alveolar epithelial cells, infiltration of alveolar macrophages, peribronchial and perivascular lymphoid aggregates. Hyperplasia of bronchiolar epithelium in focal areas and desquamated bronchial epithelial cells were found as constant lesion by the end of 12th week in a dose dependent manner. These changes were mild in OS ameliorated groups when compared to corresponding lead treated groups. Testis showed interstitial edema, thickened basement membrane of seminiferous tubules, disruption of basement membrane, separation of seminiferous tubular epithelium from basement membrane, shrinkage of tubules, basement membrane with spermatogonial cells, desquamated seminiferous tubular germ cells, disappearance of Leydig cells and MNC infiltration in interstitium and presence of multinucleated cells in seminiferous tubules were conspicuous by the end of 12 weeks. Pancreas revealed degenerated islets of Langerhans, congested, thickened and hyalinized blood vessels, hyperplasia of ductular epithelium, periductular MNC infiltration, necrosis and atrophy of islets, vacuolar degeneration in acinar epithelium and MNC infiltration between acini, interlobular hemorrhages, extensive interlobular fibroblast proliferation and severe necrosis of islets of pancreas were more conspicuous in majority of animals in lead treated groups (Groups II & III) when compared to control group by the end of 12th week. These changes were with reduced intensity in OS ameliorated groups. Thyroid of lead treated rats revealed dose dependent changes like hemorrhages, severe desquamation of acinar epithelial cells, and complete absence of colloid in acini, disruption of acini with absence of colloid and atrophy of acini were noticed throughout experimental period. Very mild changes were noticed in Ocimum treated groups. . Skin of lead treated groups revealed mild MNC infiltration in dermis, mild edema in dermis and thinning of epidermis, vacuolar degeneration of epidermal cells and sebaceous glands, atrophy of hair follicles and hyperplastic sebaceous glands were more conspicuous in a dose dependent manner. These changes were in mild intensity in OS ameliorated groups. Histochemically more intense alkaline phosphatase reaction was noticed in epithelium and basement membrane of proximal convoluted tubules of kidney and interstitial tissue and basement membrane of seminiferous tubules and less reaction in spermatogonial cells of testis of lead fed groups in a dose dependent manner. Where as a dose dependent decrease in alkaline phosphatase reaction was noticed in OS ameliorated groups. Immunohistochemistry was done to detect apoptotic bodies in liver and kidney by using the monoclonal antibodies against BAX and BCl2 antigens. The reaction was more pronounced in the hepatocytes around central veins and peripheral hepatocytes of hepatic lobule of liver, where as in kidney, the reaction was found in epithelial cells of PCT of lead treated groups in a dose dependent manner. The reaction was less in OS ameliorated groups. Ultra structurally, kidneys of lead treated groups revealed swollen mitochondria with degeneration, fragmented endoplasmic reticulum, increased number of lysosomes, clumping of nuclear chromatin and Intranuclear electron dense lead inclusions with different sizes. Where as in liver, increased number of darkly stained lysosomes, swollen and decreased mitochondria and less endoplasmic reticulum, clumping of nuclear chromatin in hepatocytes. Swollen and vacuolated vascular endothelial cells, degenerated myelin sheath, decrease in mitochondrial density margination and clumping of nuclear chromatin in brain were noticed in dose dependent manner. Residual analysis of blood and tissue samples (liver, kidney, muscle and testis) of all treated groups showed highest lead concentration in kidney followed by liver, testis and muscle. The highest concentration of lead was found in group II then followed by group III and other ameliorated groups as a dose dependent manner. A non significant reduction was noticed in OS ameliorated groups. Epidemiologically, the highest lead residuals were found in blood and milk samples of cows found near batteries factory followed by lamco factory followed by Neutrine factory, Sugar factory (Puttur) and sugar factory (Chittoor). Similarly the highest lead concentration was found in kidney followed by liver and muscle of cattle reared in and around Renigunta. Where as the highest concentration was noticed in kidney, liver, muscle and testis of sheep reared in Chittoor followed by Tiruapti urban area.