Browsing by Author "SRILATHA, Ch (Major)"
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ThesisItem Open Access CLINICOPATHOLOGICAL STUDIES ON DERMATOSES IN DOGS(Sri Venkateswara Veterinary University, TIRUPATI – 517 502,A.P, 2010-12) SRIDHAR, R; SRILATHA, Ch (Major); ANNAPURNA, P; SURESH KUMAR, R.VABSTRACT : The present study was conducted on 90 dogs with dermatological problems presented at the Veterinary polyclinics and private pet clinics. Skin biopsies obtained from 44 dogs with non-neoplastic lesions suffering from recurrent dermatoses were subjected to clinical, hematological, biochemical, cytological and histopathological studies. Excision biopsy samples were obtained from 46 dogs with neoplastic lesions. Based on clinical observations the lesions were categorised into 7 groups namely, demodicosis (29.54%), malasseziasis (13.63%), flea allergic dermatitis (15.90%), seborrhoea (6.81%), hot spots (4.54%), hypothyroidism (9.09%) and miscellaneous (20.45%). Hemoglobin values were significantly decreased in dogs with demodicosis, hot spots and hypothyroidism. Absolute neutrophil count was significantly high in dogs with demodicosis. Except in the dogs with seborrheic dermatitis, a significant decrease in the glucose values were observed between healthy dogs and dogs with dermatological problems. The serum albumin was significantly low in dogs with demodex and hot spots. Albumin globulin ratio was significantly low in dogs with Demodex and flea allergic dermatitis. Microscopically epidermal hyperplasia, hyperkeratinisation, parakeratosis, perifolliculitis and furunculosis were seen in demodicosis. In malasseziasis, hyperkeratosis, acanthosis, spongiosis, scalloping of epidermis, formation of retepegs and increased collagen in dermis were noticed. In seborrhoea, acanthosis, parakeratosis and plugging of follicles with keratin were seen. Infiltration of neutrophils and mild subepidermal infiltration around blood vessels throughout the dermis were seen in flea allergic dermatitis. In hot spots, necrotic debris with neutrophils in epidermis and edema along with infiltration of neutrophils in the dermis were noticed. Epidermal atrophy, melanosis of epidermis, atrophy of sebaceous glands, hyperkeratosis, telogenisation of follicles or atrophy of follicles, dysplasia of follicles in few dogs were noticed in hypothyroidism. Edematous sweat glands, hyperkeratosis were seen in the dogs in the miscellaneous group. The pattern analysis revealed perivascular pattern (37dogs), intraepidermal pustular pattern (12), subepidermal pustular pattern (4), follicular pattern (5 dogs), diffuse pattern (3 dogs), Panniculitis (1) and atrophic pattern in 16 dogs. Perivascular pattern was the most common pattern noticed in the biopsies. Dogs with demodicosis revealed perivascular pattern, follicular pattern, subepidermal pustular pattern and diffuse pattern. Skin biopsy specimens of dogs with malasseziasis revealed perivascular pattern, intraepidermal pustular pattern, and subepidermal pustular pattern. Histopathological analysis of neoplastic lesions in 46 dogs revealed many types of tumours of which perianal tumour was the most commonly occurring tumour (21.73%) followed by TVT (10.86%), basal cell carcinoma (10.86%), ductular adenocarcinoma of sweat gland (6.52%), squamous cell carcinoma (4.34%), mast cell tumour (4.34%), sebaceous adenoma (4.34%) etc.ThesisItem Open Access HISTOPATHOLOGICAL, IMMUNOHISTOCHEMICAL AND ULTRASTRUCTURAL STUDIES ON ENDOMETRIAL BIOPSIES IN INFERTILE BUFFALOES(Sri Venkateswara Veterinary University, TIRUPATI – 517 502,A.P, 2010-01) SAMATHA, V; SRILATHA, Ch (Major); ANJANEYULU, Y; SURESH KUMAR, R.VABSTRACT : Buffaloes play an important role in dairy developmental programmes of our country. In spite of tremendous advances in veterinary medicine, during the past 50 years, the infertility in bovines remained as almost important economic factor. Poor reproductive efficiency is one of the major problems faced by buffalo breeders. Among various pathological conditions of female reproductive tract, inflammation and unfavourable uterine environment is the most important etiological factor for infertility in bovines. Endometrial biopsy examination is the most reliable diagnostic tool for veterinarians to identify the nature of infertility and so aid in reproductive herd health programme. It can be used to identify histopathological changes of endometrium and thus serve as a basis for prognostic evaluation of infertile animals. Diagnosis of sub clinical cases is possible only by histopathological examination of uterine biopsy samples. The isolation of microorganisms along with histopathological studies of endometrial biopsy permits more accurate assessment of reproductive prognosis and rational treatment of reproductive breeding of animals. For the present study, one hundred and ten endometrial biopsies samples were collected from infertile buffaloes after thorough rectal examination and in absence of palpable uterine abnormalities. Biopsies were collected aseptically by Albuchins uterine biopsy catheter after induction of epidural anaesthesia under sterile conditions. Uterine aspirate was collected aseptically and carefully by sterile uterine catheter for cytological examination. Histopathological, cytological, bacteriological, immunohistochemical and electron microscopic studies were carried out on uterine biopsies. Histopathologically, the lesions observed were acute, subacute, chronic, chronic suppurative and chronic catarrhal changes in 14.54%, 34.54%, 46.37%, 1.82% and 2.73% biopsies respectively. Acute endometritis cases revealed severe congestion of endometrial blood vessels, stromal edema, degenerative changes in luminal epithelium, focal areas of denudation of epithelial lining and infiltration of polymorphonuclear cells and few lymphocytes. Endometrial cytological smears in acute endometritis cases revealed more number of neutrophils. Moderate infiltration in sub epithelial zone of stratum compactum, moderate periglandular fibrosis, hypertrophy of blood vessels, cystic dilation of endometrial glands in addition to stromal edema and glandular edema were noticed in sub acute endometritis. Lymphocytes and polymorphonuclear cells were observed in addition to epithelial cells in endometrial cytology of sub acute endometritis cases. Diffuse infiltration of lymphocytes, plasma cells and macrophages in stratum compactum and stratum spongiosum, thick fibro cellular endometrial stroma and gland site masses with severe periglandular fibrosis were observed in chronic endometritis cases. Severe proliferation of fibroblasts, glandular sclerosis and hyalinization of blood vessels were also noticed in chronic endometritis. Few chronic endometritis cases revealed cystic dilatation of endometrial glands and glandular hyperplasia in other non affected glands. Endometrial cytology of chronic endometritis cases revealed lymphocytes, few plasma cells in addition to epithelial cells and mucin strands. Chronic suppurative endometritis cases revealed periglandular fibrosis and infiltration of more polymorphonuclear cells in stratum compactum and stratum spongiosum. More number of degenerating polymorphonuclear cells in addition to colonies of cocco-bacilli bacteria were observed in endometrial cytological smears of chronic suppurative endometritis. Chronic catarrhal endometritis cases revealed catarrhal changes in luminal epithelium and thick connective tissue stroma in endometrium. Bacteriological examination of endometrial biopsy samples yielded majorily Salmonella (32.5%) followed by E.coli (23.75%), Staphylococcus (28.75%), Pseudomonas (8.75%), Klebsiella (2.5%), Pasteurella (2.5%) and Streptococci (1.25%). Immunohistochemical studies of chronic endometritis biopsies revealed more number of CD3 positive cells (pan T lymphocytes) in stratum compactum. Six chronic endometritis biopsies revealed CD138 positive cells (plasma cells) in endometrial stroma. Scanning electron microscopic examination of uterine biopsies collected from normal endometrium revealed surface epithelial cells with few ciliated and non ciliated cells. Loss of cilia and microvilli of surface epithelium was observed in acute endometritis cases .Sub acute endometritis cases revealed rod shaped bacteria adhering to surface epithelial cells and damaged epithelial cells with loss of microvilli. Destruction of surface epithelium and glandular structure leaving hole like spaces and fibrosis with thin long reticulin fibers were noticed in chronic endometritis cases.ThesisItem Open Access PATHOLOGICAL STUDIES ON THE EFFECT OF PHYTOGENIC SILVER NANOPARTICLES ON AFLATOXICOSIS IN BROILERS(Sri Venkateswara Veterinary University, TIRUPATI – 517 502,A.P, 2011-10) RAVI BABU, G; SRILATHA, Ch (Major); SUJATHA, K; SRINIVASULU, D; ADILAXMAMMA, KABSTRACT : Aflatoxin is a common contaminant in a variety of tropical and subtropical feeds and feed stuffs which are being used in various livestock and poultry feed rations regularly. To address the problem of aflatoxicosis, the present study was taken up to evaluate the ameliorating effect of phytogenic (Zea mays) nanosilver on aflatoxicosis in broilers. Aflatoxin was produced from a culture of Aspergillus parasiticus grown on rice. The yield of aflatoxin obtained was 30 mg per kg rice. Silver nanoparticles were synthesized through biological method using maize (Zea mays) leaves. A total of 120 male broiler chicks were divided into six groups of 20 birds each. Group I served as basal feed control, groups II and III served as phytogenic Nanosilver controls at the dose of 250 ppm and 500 ppm respectively given in water. Group IV aflatoxin was B1 at the dose of 2 ppm in the feed. Groups V and VI were given aflatoxin at 2 ppm in feed along with 250 ppm and 500 ppm of phytogenic Nanosilver respectively in water. The study was carried for six weeks. Six chickens were sacrificed from each group at fortnight interval, to study the objectives of finding out the effect of aflatoxin on hemato-biochemical, oxidative damage, gross and histopathological, antioxidant profile in liver, ultra-structural changes in liver, residual aflatoxin content in liver and ameliorating effect of nanosilver against aflatoxicosis Aflatoxin treated birds were dull and showed reduced weight gain and feed consumption. The mortality in aflatoxin and aflatoxin with 250 and 500 ppm nanosilver was to the extent of 30%, 20% and 10% respectively. The mean body weight gain (g) of aflatoxin treated group showed a significant (P< 0.05) decrease in comparison with other groups. Treatment with nanosilver in groups V and VI showed significant (P<0.05) increase compared to aflatoxin group. The feed consumption (kg) values of aflatoxin fed groups II, V and VI were significantly (P<0.05) lower when compared to groups I, III and IV. The feed conversion ratio (FCR) of group II was significantly (P<0.05) higher compared to group I. Group V supplemented with nanosliver 250 ppm, showed higher FCR values than groups I, III, IV, VI and lower than aflatoxin group II. The mean PCV (%) value of group II showed a significant (P<0.05) reduction when compared to other groups except group V with which it was comparable. The mean Hb (g%) levels of group II showed a significant (P<0.05) reduction when compared to the other groups except group V. The mean heterophil and eosinophil count of group II, group V and group VI showed a significant (P<0.05) increase and the mean lymphocyte count showed a significant (P<0.05) decrease when compared to group I, III and IV. The mean basophil and monocyte count of all the groups showed no significant difference between themselves. The details of total protein (g%) of group II showed a significant (P<0.05) reduction compared with other groups. The mean total protein concentration of group VI was significantly higher than group IV and significantly lower than groups I, III and IV. The mean value of albumin of group II showed a significant (P<0.05) reduction compared with other groups. In group V and VI the values were significantly higher than group II and significantly lower than groups I, III and IV. The mean AST activity (IU/L) of group II, V and VI birds showed a significant (P<0.05) increase when compared to other groups. However, there was no significant difference between groups I, III and IV. The mean TBARS concentrations in group II, V and VI which were comparable between themselves were significantly (P<0.05) higher compared to group I, III and IV. The mean values of GSH (g/g) of liver of group II, V and VI which were comparable between themselves were significantly (P<0.05) lower compared to groups I, III and IV. Significant gross lesions were observed only in aflatoxin fed group birds. Lesions included pale to yellowish enlarged liver with hemorrhages, pale enlarged kidney, pale heart, hydro pericardium, enlarged and congested bursa, catarrhal changes in intestine and brain with mild congestion. In nanosilver treated birds (Group V and VI) similar changes were noticed with mild degree. Microscopically, the livers of aflatoxin fed birds revealed moderate to severe diffuse microvesicular to macro vesicular fatty changes, sinusoidal hemorrhages, mild periductular fibrous tissue proliferation, acinar pattern of the hepatic cell, with ductular infiltration by mononuclear phagocytes and bile ductular hyperplasia which were pronounced with marked congestion in 4th week. In addition, perivascular infiltration of diffuse mononuclear and eosinophilis, dissociation of hepatic cells and preneoplastic changes were also conspicuous. At the end of 6th week, liver revealed marked fatty degeneration besides multiple areas of mononuclear, eosinophilic infiltration and formation of eosinophilic granuloma. In group V at the end of 2nd week, liver revealed mild fatty changes, mild fibro blast proliferation and mild eosinophilic infiltration and regenerative changes. Moderate degeneration of hepatic cells and mild eosinophilic infiltration were observed at the end of 4th week. However, hepatic cell degenerative changes and eosinophilic infiltration were markedly reduced. Group VI showed focal mononuclear cell infiltration, congestion and fatty changes at the end of 2nd week. At the end of 4th week, liver showed mild fatty changes with mononuclear phagocytic infiltration, eosinophilic infiltration, hyperchromatic nuclei and focal aggregation of mononuclear cells. At the end of 6th week besides acinar arrangement of hepatic cells, thickened blood vessels and fibrous tissue proliferation, periportal sinusoids dilatation, with congestion, and hyperplasia of bile duct were observed. The kidney sections of the birds in group II showed degenerative and desquamatory changes in tubular epithelium, hyperplasia and hypertrophy of glomerulus, inter tubular hemorrhages and thickening of bowman’s capsule by 2nd week. These changes were pronounced by 4th week. In 6th week severe congestion, inter tubular hemorrhages and focal areas of necrotic changes in tubular epithelial cells, and few areas of atrophied glomeruli in addition with the above changes were noticed. In group V and VI similar changes were noticed in kidneys like that of aflatoxin fed group. In group II, sections of bursa of Fabricius revealed severe depletion and mild edema in the bursal follicles, small to big sized interfollicular and intrafollicular cystic spaces and lack of differentiation between cortex and medulla, vacuolation in lining epithelium (plicea) mild follicular hemorrhages and interfollicular fibrosis at the end of 2nd week. A mild metaplastic change in lining epithelium and sub epithelial infiltration was observed at the end of 4th week. At the end of 6th week foam cells, inter follicular cystic spaces, acinar formation, inter follicular fibrosis, and hyperplasia in plical epithelium were observed conspicuously. In group V and VI, mild depletion was evident. Birds in group II spleen revealed mild depletion of lymphocytes, congested blood vessels and mild engorgement of red pulp at the end of 2nd week. While moderate depletion was observed towards the end of 4th and 6th week of experiment. In groups V and VI, mild depletion of lymphocytes were observed. Thymus revealed severe cortico-medullary hemorrhages in group II,Vand VI birds throughout the experimental period. Pancreatic sections revealed mild congestion, decrease in the size of islets and focal areas infiltration, thickening of the pancreatic duct, individualization of acinar epithelial cells and focal mononuclear infiltration, pancreatic ductular hyperplasia and mild inter acinar fibrosis were observed conspicuously in aflatoxin fed birds. In nanosilver treated group V and VI, mild vacuolation and focal necrotic changes in acinar epithelium and thickened blood vessels were observed, congested blood vessels, thickening of pancreatic duct, inter acinar fibrosis and focal areas of mononuclear infiltration were observed. In group II, heart sections revealed sarcolysis, mild hemorrhages and mild infiltration of mononuclear cells and mild edema at the end of 2nd week, which were pronounced with marked sarcolysis 4th week. Sarcolysis, congested blood vessels, edema, perivascular fibrous tissue proliferation with intermuscular infiltration of mononuclear cells were observed by the end of 6th week. Groups V and VI revealed sarcolysis, focal areas of mononuclear cell infiltration and inter muscular hemorrhages in 2nd week. Similar changes were observed in treated birds in 4th week and 6th week. In group II birds brain sections revealed proliferation of capillaries, satellitosis, gliosis, central chromatolysis and demylinating changes were seen in cerebrum. In cerebellum grouping and atrophy of the purkinje cells and spongiosis were noticed. Similar changes were noticed in nanosilver treated group V and VI In lung, extensive hemorrhages, congestion, subepithelial eosinophilic infiltration in bronchi, peribronchiolar and mononuclear infiltration, bronchiolar epithelial hyperplasia and perivascular edema were observed throught the experimental period in aflatoxin fed group II birds. In nanosilver treated groups (Group III and IV) mild congestion, hemorrhages and mild mononuclear cell infiltration were noticed by 6th week. In nanosilver treated birds (group V and VI) edema, thrombus formation, severe infiltration of mononuclear cells and thickening of blood vessels were observed. In Afltaoxin fed group birds, intestine revealed desquamation of villus epithelium, severe infiltration of lymphocytes in mucosa, hyperplasia of villus epithelium and focal area desquamation of epithelium by 2nd and 4th week. While pronounced disruption of villus epithelium and increase in number of goblet cells were observed in birds treated for 6 weeks. In group V, lamina propria was infiltrated with lymphocytes and hyperplasia of villus epithelium were observed throughout the experimental period. In group VI, hyperplasia of villus epithelium and severe desquamation were noticed in higher dose of nanosilver treated birds throughout the experimental period.ThesisItem Open 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, DABSTRACT: 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.ThesisItem Open 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, DABSTRACT : 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.