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ThesisItem Open Access PATHOLOGICAL STUDIES ON ENDOSULFAN TOXICITY IN RATS AND ITS AMELIORATION WITH OCIMUM SANCTUM(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2008-11) BHARATH, B.K; ANJANEYULU, Y(MAJOR); Srilatha, ChABSTRACT : 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.ThesisItem Open Access STUDIES ON PATHOLOGY OF INDUCED FENVALERATE TOXICITY IN RATS WITH SPECIAL REFERENCE TO NEUROENDOCRINE SYSTEM AND ITS AMELIORATION WITH WITHANIA SOMNIFERA (ASHWAGANDHA)(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2008-11) AMARAVATHI, P; SRILATHA, Ch(MAJOR); RAMADEVI, V; SURESH KUMAR, R.VABSTRACT : The effects of pesticides have been recognized as a serious public health concern during the past decades. Pyrethroids and organophosphates have a wide spectrum of insecticidal potency and produce acute toxicosis in vertebrates manifested by the inhibition of acetylcholine esterase (Mohamed et al. 1993). Fenvalerate is one of this groups which is potent insecticide that has been in use since 1976. Over the last 20 years, large amounts of formulated fenvalerate, approximately 1000 metric tones per year have been used as an agricultural pesticide (WHO, 1992). Although fenvalerate considered having low acute toxicity to mammals, it is having severe neurotoxic effects and estrogenic activity causing endocrine disorders. Hence fenvalerate is enlisted as one of the neuroendocrine disrupting chemicals (EDCs) and has aroused world wide concern. EDCs are known to act at multiple sites through multiple modes of action, but the mechanism of action is poorly understood (Junhe et al. 2006). Persual of literature revealed limited information regarding toxicopathological effects of fenvalerate with special reference to neuro endocrine system (Mani et al. 2002). Organochlorines and organophosphates have been proved to protect oxidative stress and very few reports are available regarding involvement of reactive oxygen species (ROS) in pyrethroid toxicity. Beneficial influence of vitamin E and vitamin C are well documented in reducing the harmful effects of fenvalerate and limited information is available regarding the herbal products in ameliorating the pyrethroid toxicity. Keeping in view of its wide spread use, the present study is taken up to investigate toxicopathological manifestations. The present study was carried out by procuring 144 female rats and were randomly divided into eight groups consisting of 18 rats in each group. Fenvalerate (20% EC) gavaged per orally using ground nut oil as vehicle @ 42.5, 21.25 and 10.125 mg / kg b.wt. to groups II, III and IV respectively and ashwagandha @ 200 mg / kg b.wt. in distilled water was given along with fenvalerate to group V, VI and VII for 6 weeks to study ameliorative effects. Group I and VIII kept as control for toxin and ashwagandha fed groups respectively. Six rats from each group were sacrificed at fortnight interval. Dullness, depression, pawing, burrowing, hyper excitability, piloerection, salivation, lacrimation, clonic seizures and whole body tremors were the main clinical signs in all toxin fed and ameliorated groups. Significant (P<0.05) decrease in the body weight gains was observed in higher dose levels of fenvalerate fed groups (group II and III) when compared to control groups (I &VIII). In ameliorated groups no significant difference was observed when compared to corresponding toxin treated groups. Significant (P<0.05) decrease in TEC, PCV, haemoglobin and TLC values was observed in all fenvalerate treated groups when compared to control groups. There was no significant difference among treated groups and ameliorated groups. There was no significant difference in ALC and ANC values. Significant (P<0.05) decrease in serum total protein, serum cholesterol and serum acetyl choline esterase and significant increase in serum AST, ALT, serum glucose and creatinine levels were observed in all fenvalerate treated groups There was no significant difference between toxin treated groups and ameliorated groups. But in group VII significant improvement was observed when compared to corresponding toxin treated group. There was significant (P<0.05) decrease in T3, T4 and E2 levels in all fenvalerate treated groups when compared to control groups and group VII. In between groups significant variation was observed. Significant improvement was observed only in lower dose levels of toxin with ameliorating agent (group VII). Significant decrease in HA titers and DNCB dermal sensitivity was observed in all toxin fed groups and indicating decrease in cell mediated immunity and humoral immunity. Grossly mild changes were noticed only in liver, lung, spleen and Brain. Histopathologically lesions were prominently noticed in kidney followed by liver, lung, brain, ovary, uterus, heart, spleen, lymph node and intestine in all toxin fed groups. In kidneys congestion, edema, degenerative changes, intertubular haemorrhages, mild fatty changes in tubular epithelial cells were observed. Glomerular atrophy and fibrous tissue proliferation around glomeruli were consistently recorded in fenvalerate treated groups in a dose dependent manner. Cystic tubules and glomeruli were observed in majority of higher doses of fenvalerate treated rats by the end of experiment. In ameliorated groups also similar changes were observed except in group VII where mild congestion and mild degenerative changes were observed. Liver revealed congestion, degenerative changes, haemorrhages, mild fatty changes, infiltration of MNCs, loss of hepatocytes and proliferation of bile ducts in all fenvalerate treated groups.. Apart from prominence of lesions in group II, other changes observed were individualization of hepatocytes, perivascular infiltration of MNCs and hyper chromatic nuclei in some hepatocytes. In ameliorated groups also similar changes were observed except in group VII where mild sinusoidal haemorrhages and mild degenerative changes were observed. In lungs moderate to severe congestion, edema, perivascular and peribronchiolar infiltration of mononuclear cells and hypertrophy of lymphoid follicle was observed. In many rats areas of emphysema, thickened alveolar septa due to edema, infiltration of MNCs, RBCs, macrophages and few plasma cells, hyalinised blood vessels and giant cells in alveolar lumen was observed prominently in higher doses of fenvalerate treated groups (group II and III). In lower doses of toxin (group IV) only mild pneumonic changes were observed. In ameliorated groups also similar changes were observed except in group VII where mild congestion with focal infiltration of MNCs was observed. Histopathologically cerebrum revealed congestion, submeningeal haemorrhages and demyelinating changes in all fenvalerate treated groups. In addition gliosis, central chromatolysis and proliferation of capillaries were observed in cerebrum of group II and III rats. Sub meningeal haemorrhages, focal loss, shrinkage, central chromatolysis of purkinjee cells in the purkinjee cell layer and mild congestion and haemorrhages in granular layer of cerebellum were observed more conspicuously in group II and III rats. In ameliorated groups also similar changes were observed except in group VII where only mild changes were noticed. Microscopic examination 0f heart revealed mild to moderate haemorrhages and congested vessels and mononuclear cell infiltration in between cardiac muscle fibers. In addition to above lesions, sarcolysis along with eosinophilic cellular infiltration were observed conspicuously in higher doses of fenvalerate treated groups. Similar changes were observed in ameliorated groups (group II and III). Microscopic lesions in intestine were not characteristic in toxin treated rats except moderate increase in number of goblet cells, necrosis and hyperplasia of intestinal epithelial cells in group II and III. Histopathological examination of spleen revealed congestion, focal areas of hemorrhages, mild lymphocytolysis and engorgement of red pulp with erythrocytes in group II and III. In lymph node congestion of blood vessels and mild to moderate lymphocyte depletion was observed group II and III. In pancreas degenerative changes in islets of Langerhans, serous acini, necrosis, congestion, haemorrhages, infiltration of MNCs loss of acini with fibrous tissue proliferation was observed in group II and III. In adrenals sinusoidal and severe medullary haemorrhages and degenerative changes in medulla were observed in all fenvalerate treated groups. In ameliorated groups similar changes like those of corresponding toxin treated groups were observed in intestine, spleen, lymph node, pancreas and adrenals. In ovary severe haemorrhages, hyaline fibrosis, increased number of atretic follicles, degenerated follicles and decreased number of primordial follicles were observed in all toxin treated groups in dose dependent manner. Similar changes were observed in ameliorated groups. Uterus revealed congested blood vessels, haemorrhages, hyperplasia of endometrial epithelium, periglandular edema and eosinophilic cell infiltration with few plasma cells, lymphocytes in sub mucosa and atrophy of endometrial glands and periglandular fibrous tissue proliferation in all fenvalerate treated groups in dose dependent manner. In ameliorated groups similar changes were observed. Ultra structurally dose dependent marked distortions in membrane, irregular shape with many small shallow pits and small perturbations on the surface were noticed by the end of 6th week in all toxin treated groups when compared to control. In ameliorated groups mild distortion was observed. In liver swollen nucleus, margination of chromatin material, thickened nuclear membrane, vacuolation of nucleoplasm, vacuolation of cytoplasm, uniform reduction of size of mitochondria and mild fatty deposition were noticed in all toxin treated groups in dose dependent manner. In ameliorated groups mild to moderate changes were observed when compared corresponding toxin treated groups.ThesisItem Open Access PATHOLOGICAL STUDIES OF EXPERIMENTALLY INDUCED COMBINED (ISN + RIF) DRUGS TOXICITY IN RATS AND PROTECTIVE EFFECTS OF VITAMIN-E AND WITHANIA SOMNIFERA(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2008-10) ARUNDHATHI, S; ANAND KUMAR, A(MAJOR); ANJ ANEYULU, Y; GOPALA REDDY, AABSTRACT: A total of 24 healthy male Wistar rats (age 60-100 days, weighing 175-260mg) procured for the present experimental study and the experiment was carried out according to the guidelines and prior approval of Institutional Animal Ethics Committee. Animals were divided into four groups consisting of 6 in each group. The experimental study was designed as follows:- Group I-Control, Group 11-ISN@SOmg/kg body wt + RIF@lOOmg/kg body wt once daily orally for 21 days, Group 111-ISN@SOm&g body wt + FUF@lOOrng/kg body wt once daily orally for 21 days + Withania sornnifira@lOOOmgkg feed daily for 21 days, Group N-ISN@SOm&g body wt + RIF@lOOmgkg body wt once daily orally for 21 days + vitamin E@300mg/kg feed daily for 21 days. Whole blood and serum was collected 4 times at weekly intervals (0, 7, 14 and 21 days) for estimation of various haematological and biochemical parameters and analyzed statistically. Haematological studies revealed that overall means of PCV, Hb and TEC were significantly (P<0.05) reduced and TLC was increased significantly (Pc0.05) in group I1 in comparison to other groups. The biochemical assays showed a significant (Pc0.05) reduction in total protein, HDL cholesterol, while significant (Pc0.05) increase in AST, ALT. GGT, total cholesterol and triglycerides in group 11. The - tissue enzyme assays revealed a significant (P<0.05) increase in TBARS and significant (P<0.05) decrease of GSH values in group 11. The ameliorative groups 111 and IV showed significant improvement in all parameters in comparison to group 11. The gross pathological changes in the group I1 animals included mild hepatomegaly. rounded borders, congestion and in heart petechiae and ecchymotic haemomhages with focal haemorrhages on kidney. The histopathological changes in group I1 revealed enlarged portal tracts with infiltration of lymphocytes, severe sinusoidal congestion and fatty change in perigortal areas. Hepatocytes revealed degeneration and moderate to severe necrosis. Heart revealed disruption and separation of muscle fibres, endocardid haemorrhages, moderate infiltration of lymphocytes and degeneration of muscle fibres, necrosis and fragmentation. In kidney glomeruli revealed reduction in glomerular space, vacuolation in few glomeruli and degeneration with extensive haemorrhages in interstitial tubular spaces, tubular epithelial cells showed fatty change with disrupted tubules and focal areas of tubular atrophy. Brain revealed congestion, haemorrhage and infiltration of mononuclear cells in meningeal vessels surrounding areas fatty change and increased cellularity in granular cell layer of cerebellum. Group 111 and IV animals revealed mild infiltration of mononuclear cells in and around portal areas with mild sinusoidal congestion. Structural details were intact and mild fatty changes were evident. Heart revealed mild range of degeneration and disruption of muscle bundles. Kidney showed mild haemorrhages in interstitial spaces and normal glomerular space. Brain revealed decreased cellularity and mild congestion in and around the meningeal areas. The present study concluded as follows: I. The changes resulted by the antitubercular drugs in the haernatology can be attributed to the hepatic damage. 2. The biochemical changes might be attributed to the toxic metabolites of the isoniazid and rifampicin. 3. The gross and the histological changes in the study might be due to the toxic reactive metabolites of the drugs which binds to cellular macromolecules and release or form toxic free radicals inturn caused the tissue damage. 4. Supplementation of Withania somnifera (1%) resulted in significant improvement in the haematological and biochemical parameters might be due to the antioxidant, antistress, liver tonic and anti-inflammatory properties. 5. Supplementation of vitamin E (0.3%) resulted in significant improvement in the haematological and biochemical parameters which can be attributed to the antioxidant and anticholesterimic effects. 6. Elevation of tissue TBARS and reduction of GSH in the study might be because of increased lipid peroxidation due to toxic reactive metabolites of the drugs. The present study indicated that Withania somnifera @ 1OOOmgkg feed and vitamin E @300mg/kg feed were effective in counteracting the toxic effects of the antitubercular drugs but not up to the required levels. Keeping this in view, Wer studies can be advocated using different dose rates and different routes of administration to overcome the affects of antitubercular drugs which would be beneficial for conservation of rare species on the earth.ThesisItem Open Access PATHOLOGY OF NEOPASMS IN DOGS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2008-10) CHANDRAVATHI, T; ANJANEYULU, Y(MAJOR); ANAND KUMAR, A; NARASIMHA REDDY, YABSTRACT : Cancer is the most urgent problem of the contemporary medical field. In most of countries it is the second leading cause of the death, next to the cardiovascular disease. Neoplasm is a leading cause of death in all species. In animals dogs have highest incidence. About 45% dogs reaching middle age (6-7 years) will either develop a tumour, suffer medical complication as a result of tumour, or die as a result of neoplastic disease so there is need to emphasise on canine neoplasms. The present study was carried out to know age, breed and sex wise incidence of canine neoplasms in and around Hyderarabad and samples were collected from different hospitals after surgical incision. Immunohistochemistry was carried out to know the proliferating activity in different tumours. A total of 68 samples were collected, out of which 31 (45.59%) were benign and 37 (54.41%) were malignant tumours. Tumours were classified into epithelial 40 (58.83%), mesenchymal 20 (29.40%), round cell 5 (7.36%) and mixed tumours 3 (4.41%). The highest risk of development of various tumours was found in the age group of 7-9 years, followed by 4-6 years, above 9 years and below 3 years and the incidence was 29 (42.65%), 20 (29.41%), 19 (27.94%) and 1 (1.47%) respectively. The frequency of occurrence of neoplasms was slightly higher in females 38 (55.88%) compared to the males 30 (44.12%). Among the breeds affected Pomaranian breed represented more with 18 (26.50%) followed by non-descriptive 16 (23.52%), German Shepherd 16 (23.52%), Labrador 6 (8.82%), Doberman 6 (8.82%) Dachshund and Rottweiler 2 (2.94%), each one of Collie and Great Dane (1.47%). The organ wise incidence of tumours, included skin and mesenchymal tumours 29 (42.65%), mammary tumours 17 (25.0%), joint and bone tumours 9 (13.23%), vaginal tumours 5 (7.35%), testicular tumours 4 (5.88%), vulva and penis 3 (4.41%), ovarian tumours 2 (2.94%) and each one of transitional cell carcinoma and prostatic carcinoma (1.47%). Benign tumours include, fibroma 3 (4.42%), sebaceous gland adenoma 2 (2.94%), perianal gland adenoma 2 (2.94%), papilloma 2 (2.94%), melanoma 2 (2.94%), canine cutaneous histiocytoma 2 (2.94%), benign mammary tumours 2 (2.94%), hemangiopericytoma 2 (2.94%), lipoma 2 (2.94%), fibromatous epulis 2 (2.94%), mammary gland adenoma 1 (1.47%), apocrine gland adenoma 1 (1.47%), anal sac adenoma1 (1.47%) myxoma 1 (1.47%) and hemangioma 1 (1.47%), kaposi like tumour 1 (1.47%), leiomyoma 1 (1.47%), fibroleiomyoma 1 (1.47%), fibromyxoma 1 (1.47%), and fibrolipomyxoma 1 (1.47%). Malignant tumours included mammary gland carcinoma 14 (20.59%), squamous cell carcinoma 3 (4.42%), transmissible venereal tumour 3(4.41%), sertoli cell tumour 3 (4.41%), fibrosarcoma 2 (2.94%), osteosarcoma 2 (2.94%), ovarian adeno carcinoma 2 (2.94%), basal cell carcinoma 1 (1.47%), malignant melanoma 1 (1.47%), seminoma 1 (1.47%), prostatic carcinoma 1 (1.47%), transitional cell carcinoma 1 (1.47%), chondrosarcoma 1 (1.47%), rhabdomyosarcoma 1 (1.47%) and lymphangiosarcoma 1 (1.47%). Oxidative stress is the central mechanism in the pathogenesis of many diseases including cancer. The present study included the estimation of TBARS, SOD, catalase, GSH and GST. The TBARS, SOD, GSH, GST and catalase levels significantly (P<0.05) increased in the tumour tissue compared to the normal tissues. The number of AgNOR’s had been associated with cell proliferation. The mean AgNOR dots varied from 2.81±1.02 to 16.89±2.58 in individual tumour. The mean number of AgNOR in benign tumours was significantly (P>0.05) lower than the malignant tumours.. The lowest AgNOR count was observed in adenoma of mammary gland (2.81±1.02) and highest in cutaneous histiocytoma (16.89±2.58). In case of mammary tumours highest AgNOR counts was recorded in anaplastic carcinoma of mammary gland (10.58±2.67). PCNA was associated with the cell proliferation in different tumours. The mean PCNA positive nuclei in tumour tissues varied from 8.23±1.25 to 437.95±8.54. The lowest PCNA index was observed in myxoma while highest PCNA index was observed in cutaneous histiocytoma. The mean number of PCNA positive nuclei in benign tumours was significantly (P<0.05) lower than the malignant tumours. Anaplastic (336.95±6.98) and solid (221.38±4.78) mammary adenocarcinomas showed highest PCNA counts than cystic papillary (93.15±7.89) and tubular adenocarcinomas (77.42±5.74). There exist a significant (P<0.01) correlation between AgNOR counts and PCNA index.ThesisItem Open Access STUDY OF PROTECTIVE EFFECTS OF ASCORBIC ACID AND SPIRULINA AGAINST TOXICOPATHOLOGY OF DOXORUBICIN IN RATS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2008-10) RAVI KUMAR, Y; ANJANEYULU, Y(MAJOR); MADHURI, D; RAJASHEKAR REDDY, AABSTRACT: Cancer is one of the major life threatening conditions in humans and animals. Unfortunately till today no drug has been found as the master in treating cancer since most of the drugs are having little or lot of side affects. Doxorubicin comes under antibiotic class of anticancerous drugs. An experiment was designed to evaluate Doxorubicin toxicity and its amelioration with ascorbic acid and Spirulina. A total of 24 healthy female Wistar rats (weighing 150-200g) procured for the present experimental study and the experiment was carried out according to the guidelines and prior approval of Animal Ethics Committee. Animals were divided into four groups consisting of 6 in each group. The experimental study was designed as follows: Group 1-Control, Group 2-Doxorubicin – Toxic control @ 2mg / kg body wt. by intravenous injection for 5 days followed by weekly once for 2 weeks, Group 3-Pre treatment with ascorbic acid @ 500mg / kg feed by enteral route for 7 days followed by intravenous injection of Doxorubicin as mentioned in group 2. Ascorbic acid supplementation was continued during these 19 days, Group 4-Pre treatment with Spirulina @ 1000mg / kg feed by enteral route for 7 days followed by intravenous injection of Doxorubicin as mentioned in group 2. Spirulina supplementation was continued during these 19days. But In group 2, 100% mortality was recorded on three days after 5th dose of doxorubicin and in group 3, 50% mortality was recorded. Hence, the experiment was terminated on day 8. Body weights were recorded on zero day and 6th day; Haematological and sero-biochemical parameters were estimated and analyzed statistically in the samples collected on 6th day. Day 1 is the day of administration of 1st dose of Doxorubicin. Body weights, heart weights and heart weight/body weight ratio were significantly (P<0.05) reduced in group 2 in comparison to other groups. Haematological studies revealed that overall means of TEC, Hb, TLC and PCV were significantly (P<0.05) reduced significantly (P<0.05) in group 2 in comparison to other groups. The biochemical assays showed a significant (P<0.05) increase in CPK, LDH, cTn, AST, ALT, Serum creatinine and BUN group 2. The tissue enzymes assays revealed a significant (P<0.05) increase in TBARS and significant (P<0.05) decrease of GSH and SOD activities in group 2. The ameliorative groups 3 and 4 showed mild to moderate improvement in all parameters in comparison to group 2. The gross pathological changes in the group 2 animals included yellowish discoloration of liver and kidney; moderate to severe congestion of heart, liver and kidney; haemorrhages in endocardium, streaks of haemorrhages in liver and petechiae in kidney.In group 2 the histopathological changes in heart revealed severe disruption, separation, degeneration, necrosis, fragmentation of muscle fibres with endocardial haemorrhages, edema, moderate focal infiltration of lymphocytes. Kidney revealed marked congestion, disrupted tubular and glomeruli architecture, shrunken / atrophied glomeruli, marked degenerative changes in tubules along with marked intertubular haemorrhages and casts. Liver revealed marked central vein and sinusoidal congestion, bile duct hyperplasia, focal areas of necrosis, karyorrhexis and marked fatty changes. Group 3 and 4 animals heart revealed moderate range of interfibrillar haemorrhages, degeneration and disruption of muscle bundles. Kidney showed mild to moderate degenerative changes. Liver revealed mild to moderate central vein, sinusoidal congestion, dilatation of sinusoidal spaces degenerative changes in the hepatocytes. The present study was concluded as follows: 1. The deleterious changes caused by doxorubicin in terms of performance and haematology can be attributed to cardiac, kidney and hepatic damage . 2. The serobiochemical changes might be attributed to the toxic metabolites of doxorubicin resulting in cellular damage. 3. The gross and the histological changes in the study might be due to the toxic reactive metabolites of the drug which binds to cellular macromolecules and release or form toxic free radicals inturn cause the tissue damage. 4. Supplementation of vitamin C (0.5%) resulted in only slight improvement in the haematological and biochemical parameters which can be attributed to the antioxidant effects. 5. Supplementation of Spirulina (1%) resulted in significant improvement in performance, haematological and biochemical parameters that can be attributed to the antioxidant, antistress and anti-inflammatory agent. The present study indicated that vitamin C @500mg/kg feed was found to be less protective affective and Spirulina @ 1000mg/kg feed offered moderate protection in counteracting the toxic affects of Doxorubicin. Keeping this in view, further studies can be advocated using different doses and different routes of administration so as to obtain best results combating the affects of anthracycline drugs which would prove beneficial for animals and humans.ThesisItem Open Access PATHOMORPHOLOGICAL STUDIES ON BUFFALO UTERUS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2008-06) ANKAMARAJU, G; Madhuri, D(MAJOR); Anand Kumar, A; Narsimha Reddy, YABSTRACT:Female genital tract of the buffalo has considerable biological and economic importance. The bovine infertility has been reported to be associated with various pathomorphological conditions in the genital organs. Reproductive disorders causes infertility ultimately leading to substantial economic losses through increased calving intervals inspite of tremendous advances in Veterinary medicine. Specific and non specific infectious agents during pre and postpartum periods frequently invades the uterus and produces metritis and endometritis leading to repeat breeding. Considering this the present study was planned to observe the incidence of gross and microscopic lesions affecting the uterus of she-buffaloes in and around Hyderabad. For the present study, three hundred and seventy five uteri of non-descript and graded murrah buffalo cows aged four to ten years with no breeding history were collected from the animals slaughtered in and around Hyderabad. Gross, histopathological, bacteriological and electron microscopic studies were carried out. Of the 375 uteri pathological lesions were observed in 145 (38.66%). The distribution of the lesions into acute, sub-acute, chronic and chronic suppurative endometritis was 11.20, 6.10, 18.10 and 3.20 percent respectively. In the present study high incidence of chronic endometritis (18.00%) was recorded. Circulatory disturbances in the mucosa were the most prominent microscopic lesions observed in acute endometritis. Severe congestion of blood vessels, enlargement of stromal cells, denudation of surface epithelium and mild to moderate infiltration of polymorphs and lymphocytes in stratum compactum was observed. In sub acute endometritis the lesions observed were extensive denudation of epithelial cells, stromal fibrosis and moderate infiltration of inflammatory cells in stratum compactum and stratum spongiosum. Severe infiltration of Polymorpho nuclear cells in perivascular and periglandular spaces of uterine stroma and moderate to severe periglandular fibrosis was observed in chronic endometritis samples. Dilation of endometrial glands and necrosis of glandular epithelium in chronic suppurative endometritis samples was recorded in addition to infiltration. 48 cases showed positive for bacterial identification. Pseudomonas, Staphylococci, Escherichia coli, Klebsiella and Proteus sp. were identified by cultural isolation. Confirmed chronic endometritis tissue was subjected to scanning electron microscopy and the electron micrographs revealed infiltration and degenerative changes in the endometrium, thickening of endometrium, presence of fibrous tissue and lack of microvilli with complete destruction of glandular structures at few places. Based on the results of present investigation it can be concluded that: 1. The incidence of chronic endometritis is high in she-buffaloes in and around Hyderabad. 2. Chronic endometritis may be a chief contributor to the incidence of repeat – breeding in buffaloes. 3. Extensive fibrotic tissue observed ultra structurally may be one of the many factors decreasing fertility in she-buffaloes.