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ThesisItem Open Access ETIOPATHOLOGY OF LAMB MORTALITY IN RAYALASEEMA REGION OF ANDHRA PRADESH(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2016-12) VENKATA RAGHAVENDRA, SANGUBOTLA; ANAND KUMAR, A(MAJOR); AMARAVATHI, P; CHENGALVA RAYULU, V; MADHAVA RAO, TABSTRACT: Present study was undertaken to study the etiopathology of lamb mortality in Rayalaseema region of Andhra Pradesh state to find out the prevalence and probable causes for lamb mortality. In an epidemiological survey, a total of 1798 lambs were recorded as dead in four districts of Rayalaseema region, of them the mortalty rate was in descending order Anatapur, Chittoor, Kadapa and Kurnool. The mortality was higher in females than males. Maximum mortality was observed in winter season and at 1-3 months age group. The mortality was majorly due to infectious origin 1533 (85.26%) followed by non-infectious origin 223 (12.4%) and miscellaneous causes 42 (2.34%). Majority of deaths were due to respiratory system affections followed by digestive system affections, multiple system involvement, combined digestive & respiratory systems, non specific affections, circulatory system, urinary system and combined respiratory & urinary system in descending order. Out of 100 faecal samples of ailing lambs examined, 68 samples were positive for different parasitic ova and oocysts. In the age group of 4-6 months it was maximum. The positive cases of parasitic ova in ailing lambs were more in females than in males. Various parasitic ova identified were eggs of Strongyle, Moniezia sp, Schistosoma sp, Haemonchus sp, Paramphistome sp, Trichuris sp and oocysts of Coccidia sp. The samples collected during the postmortem examination revealed ova of Trichuris sp, Moniezia sp, Strongyle and cysts of Balantidium sp. Out of 102 blood samples collected from ailing lambs the blood picture was decreased PCV, Hb, TEC, MCHC, neutrophils, monocytes, eosinophils and increased TLC, MCV, MCH, lymphocytes were observed when compared with apparently healthy group (n=10). Males showed increased PCV, neutrophils, eosinophils, MCV and in females increased TLC, lymphocytes, monocytes and MCHC, where as almost similar Mean±SE values of Hb, TEC were noted in both males and females. Out of 50 serum samples from ailing lambs screened, increased TP, albumin, globulin, A/G ratio, creatinine and decreased calcium, ALT were observed when compared with apparently healthy group (n=10). Increased TP, albumin, globulin, AST were reported in males whereas increased A/G ratio, calcium, ALT in females and similar values of creatinine in both sexes were noted. A total of 53 different samples collected from ailing lambs and affected organs were subjected for culture isolation of microorganisms and molecular characterization by using PCR assay. E.coli, Salmonella sp and S.aureus isolates were obtained by culture method. E.coli isolate was positive for stx2 gene, isolates of Salmonella sp for invA gene and isolates of S.aureus for nuc gene. The prevalence of gross and histopathological changes was noted from 179 dead lambs in Rayalaseema region of Andhra Pradesh during the period of study from December, 2015 to June, 2016. Lambs from Chittoor, Kadapa, Anantapur and Kurnool districts were died in descending order. The mortality was more in females than males. The highest mortality was noted in summer followed by winter. Lambs died at 4-6 months age followed by 1-3 months age, 7-9 months, 10-12 months age and <1 month age in descending order. The mortality of lambs in this region more due to multiple system affections and due to inflammatory origin majorly followed by non inflammatory origin i.e. 179 dead lambs. Inflammatory origin conditions included inflammation of multiple organs involvement was recorded to be highest. Gross lesions in respiratory system observed were tracheitis, congestion, consolidation, suppuration, aspiration, nodular growths and haemorrhages of lungs. In circulatory system, haemorrhages, congestion of coronary blood vessels, gelatinization and necrosis of epicardial fat, small pin point to streaks of pale areas on epicardium and hydro pericardium were observed in heart where as in spleen petechial haemorrhages and congestion were noted. Mild to severe enlargement, edema and hemorrhagic mesenteric lymph nodes and enlarged supra scapular lymphnode in one case were recorded. Congested cerebral blood vessels were noted. In the digestive system, rumen and reticulum filled with plenty of food material (phytobezoars), that was hard and dry, Haemonchus sp worms in abomasum, empty stomach and intestines, duodenal haemorrhages, enteritis (catarrhal and haemorrhagic), thickened intestinal mucosa, congested mesenteric blood vessels, volvulus and tape worms in intestinal portions were observed. The liver showed the lesions including congestion, necrotic areas, haemorrhages, calcification, enlarged and infarcted, cirrhotic changes, icteric patches on surface, larva migrans, abscesses and distended gall bladder. In renal system, softened (pulpy) kidneys, congestion, haemorrhages, cysts in medulla, dilatation of renal pelvis of kidneys, urinary bladder distention and urethral stricture were recorded. Three cases of Anthrax suspected lambs (confirmed by blood smear examination), One dog bite case and three abortion cases reported were not subjected to post mortem. A total of 453 tissue samples of different affected organs included lungs, liver, kidney, heart, intestine, lymphnode, spleen and brain were collected from 179 lambs that were died and subjected to histopathology. In respiratory system, pneumonia was major condition encountered and included different types, non specific pneumonia, suppurative pneumonia, bronchopneumonia, Jaagsiekte, interstitial pneumonia, fibrinous pneumonia, verminous pneumonia, aspiration pneumonia and Suppurative pneumonia combined with Jaagsiekte. In circulatory system, the lesions of heart included myocarditis, presence of Sarcocysts and parasites in myocardium. In spleen, congestion, presence of hemosiderin pigment, thickened trabeculae, degeneration and hyperplasia of splenic corpuscle and lymphoid aggregation around splenic corpuscle were recorded. In mesenteric lymphnodes the lesions included congestion, depletion of lymphoid cells, edema, haemorrhages, fibrous tissue proliferation, reactive hyperplasia, section of parasite, cyst in the vessel and abscess. A case of lymphosarcoma was also observed. Brain lesions like neuronal degeneration, vacuolation, apoptosis, neuronophagia, satellitosis and focal to diffused mono nuclear cell infiltration were observed. In kidney samples, congestion, degeneration of tubules, presence of casts, necrosis of tubules, shrunken glomeruli, fibrous tissue proliferation, cystic spaces, tubular edema, fatty change in glomeruli as well as in tubules, cloudy swelling, presence of hemosiderin pigment, infarction were noted. Accordingly the conditions classified as nonspecific nephritis, glomerular nephritis, haemorrhagic glomerular nephritis, haemorrhagic nephritis, parasitic ova and parasite in cystic fluid were recorded. In intestines congestion, haemorrhages, ulcers, fibrous tissue proliferation, degeneration of glands, various villous abnormalities were observed. The conditions were classified as catarrhal enteritis with parasitic load, nonspecific enteritis, haemorrhagic enteritis, diptheritic enteritis, necrotic enteritis. In liver sinusoidal congestion, fibrous tissue proliferation, focal to diffused necrosis, mild to moderate fatty change, degeneration of hepatocytes, bileduct epithelial hyperplasia and proliferation, calcification, presence of hemosiderin pigment were noted. Further the conditions classified as hepatitis, presence of parasitic ova, granuloma formation, abscess, haemorrhages and fibrotic nodule and bacterial colonies in portal triad.ThesisItem Open Access PATHOMORPHOLOGICAL STUDIES ON SODIUM FLUORIDE TOXICITY IN FEMALE RATS WITH SPECIAL REFERENCE TO THE REPRODUCTIVE TOXICITY AND ITS AMELIORATION(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2016-10) REDDI PRIYANKA, K; SUJATHA, K(MAJOR); SRILATHA, Ch; RANI PRAMEELA, DABSTRACT: Fluorosis is one of the most important public health hazards across the world, resulting from chronic exposure of higher amounts of fluoride. Most of the times, it is characterized by dental mottling (Dental) and crippling (Skeletal) disorders. Livestock exposed to forages and grasses contaminated with fluoride-rich industrial effluents, deep well water, feed supplements and mineral mixtures containg excess fluoride are more susceptible for fluoride toxicity. Apart from the skeletal and dental abnormalities fluoride can able to produce structural and functional alterations in many of the soft tissues like reproductive organs, liver, kidney, brain, gastro intestinal tract, thyroid and endocrine glands by impairing the antioxidant status of the body. In recent years, experimental studies pertaining to toxic effects of fluoride on reproductive organs and developmental toxicity have got great attention. In this aspect, many authors reported effects of fluoride on male reproductive system where as information regarding female reproductive system is inadequate. Provision of nutritional supplements with high antioxidant properties is the farthest objective as remedy to mitigate the complications of fluoride toxicity. Emblica officinalis (Amla) is proposed for its potent antioxidant property. ZnO nanoparticles are well documented for their multidisciplinary biological activities in the field of medicine. Hence, this present study was undertaken to investigate pathomorphological changes in the reproductive system and other soft tissues of female rats induced by fluoride and its amelioration with Emblica officinalis (Amla) and to study the efficacy of Zinc oxide nano particles coated Emblica officinalis. The present study was carried out by procuring 108 female Wister albino rats (weighing around 200 g) that were randomly assigned to 6 Groups with 18 rats in each Group. Group I served as vehicle control and received distilled water whereas animals in Groups II, III, IV, V and VI received sodium fluoride @ of 18 mg/ kg b.wt, Emblica officinalis @ of 100 mg/ rat/ day, ZnO nano particles @ of 10 mg/ kg b.wt, sodium fluoride @ of 18 mg/ kg b.wt + Emblica officinalis @ of 100 mg/ rat/ day, sodium fluoride @ of 18 mg/ kg b.wt + ZnO nano particles coated Emblica officinalis @ of 100 mg/ rat/ day respectively. Six rats from each group were sacrificed at fortnight intervals. Hematologically, significant reduction was recorded in the TEC, Hb and PCV and TLC in Group II. Serum biochemical analysis revealed significant decrease in total protein, SGPT, SGOT, cholesterol, calcium and increased creatinine, alkaline phosphatase and acid phosphatase levels in Group II. Significant increase in the concentration of TBARS, indicator for oxidative damage and decreased levels of antioxidants like catalase, SOD, GPx and GSH levels in liver, kidney, brain and uterus of Group II rats were noticed. Decreased serum Progesterone (P) and Estrogen (E2) concentration were noticed in Group II rats. Whereas, clear hematological, biochemical and hormonal improvement was noticed in Group V and VI. No alternations were noticed in all these parameters in Group III and IV. On histopathological examination, uterus of Group II rats revealed distorted lamina propria with moderate infiltration of MNCs, squeezed endometrial cells revealing large empty spaces, severe degenerative changes in endometrial glandular epithelium, periglandular infiltration of MNCs and fibrous tissue proliferation, variation in size and shape of the glands, severe hyperplastic changes with complete obliteration of glandular lumen and overall reduction in number of glands in the endometrium. Whereas in myometrium, highly distorted smooth muscle architecture with irregularly arranged funicular layers, infiltration of MNCs and eosinophils and edematous changes between muscle fibers were noticed. Uterine lumen was completely obliterated. In Group V and VI, these changes were less intense and by the end of 6th week uterus came to near normal appearance. Ovaries of Group II rats showed highly degenerated germinal epithelium, degenerated and necrosed stromal tissue with engorged blood vessels, distorted basement membrane separating the theca folliculi from the zona granulosa, vacuolated spaces in granulosa cells, severely disintegrated oocytes and nucleus in secondary follicles, increased number of atretic follicles, pyknotic follicular cells, vacuolated corpus luteum and marked reduction in the number of follicles at different developmental stages. These changes were less severe in Group V and VI. In liver, severe congestion of central vein, dilated sinusoidal spaces with congestion, degenerated hepatocytes with karyomegaly and hyperchromatic nuclei, individualization of hepatocytes, focal loss of hepatocytes with MNC aggregates and disorganization of hepatic cords, microgranuloma formation, mild to moderate vesicular fatty change, moderate fibrous tissue proliferation in portal areas, marked sinusoidal dilatation with severe congestion were observed in Group II. These changes were less intense in Group V and VI. In kidneys, extensive degenerative changes in tubular epithelial cells, severe intertubular hemorrhages and edema and proteinacious casts in tubules were noticed. Vacuolation in the tuft of glomerulus and atrophied and cystic glomeruli were evident in majority of Group II treated animals. In Group V and VI ameliorated rats, these lesions were less severe. Microscopical examination of Group II lungs showed perivascular and peribronchial infiltration of MNCs and eosinophils, moderate hemorrhages in and around the bronchiole, desquamated bronchiolar epithelium, peribronchial lymphoid aggregation and interstitial edema. These changes were less in intense in Group V and VI. Brains of Group II rats revealed shrinkage of neurons along with focal areas of demyelinating changes and spongiosis, doubling and rounding of Purkinje cells, microgranuloma formation and nerve cell with central chromatolysis and focal areas of gliosis in cerebrum. In Group V and VI ameliorated rats, these lesions were less severe. Microscopic examination of heart revealed mild to moderate hemorrhages, sarcolytic changes and infiltration of MNCs in between cardiac muscle fibers in most of the group II rats. All these changes were mild in Group V and VI. In pancreas, peripancreatic ductular infiltration of MNCs, focal loss of acinar structures exocrine duct hyperplasia and complete degeneration with atrophy of islets of Langerhans were observed in Group II rats. These changes were mild in Group V and VI. Microscopically spleens of Group II animals revealed, depletion of lymphocytes around the central artery and from germinal centers and severe engorgement of red pulp. Group V and VI animals appeared almost similar to control. No specific gross and microscopic alternations were observed in soft tissues of Group III and IV. Increased fluoride concentration was noticed in tissues like liver, kidney, brain and uterus of Group II rats whereas, significant decrease in fluoride concentration was noticed in Group V and VI. In Group II, immunohistochemistry showed increased expression of BAX marker in lining epithelium and glandular epithelium of endometrium of uterus compared to control. Decreased number PCNA positive ovarian follicles were observed in Group II compared to control. Whereas, increased number of ovarian follicles were noticed in Group V and VI. On scanning electron microscopic examination, abraded surface epithelium with distorted cuboidal shape and clumping of surface epithelial cells with decreased number of blebs and microvilli were observed in ovaries of Group II whereas, in Group V and VI, cuboidal cells of the germinal epithelium normal with significant increase in number of blebs and microvilli.ThesisItem Open Access TOXICOPATHOLOGICAL STUDY OF IMIDACLOPRID AND ITS AMELIORATION WITH VITAMIN-C IN MALE RATS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2012-09) SOUJANYA, S; LAKSHMAN, M(MAJOR); ANAND KUMAR, A; GOPALA REDDY, AABSTRACT : The present experiment was aimed to study the toxico-pathological effects of imidacloprid in male rats. Total of 48 male Sprague dawley rats were procured and divided into four groups consisting of 12 in each. The group 1 served as control, group 2 (imidacloprid toxic control at the rate of 80 mg/kg b. wt /day), group 3 was provided with vitamin C at the rate of 10 mg/kg b. wt/day, group 4 was fed with both imidacloprid at the rate of 80 mg/kg b. wt /day and vitamin C at the rate of 10 mg/kg b. wt/day. This experiment was carried out for 4 weeks. Average body weight gains were recorded at weekly intervals. A day before sacrifice the blood and serum samples were collected from six rats in each group. Tissue samples of liver, kidney, testes, brain were collected from six rats in each group on the day of sacrifice i.e. on 14th and 28th day for histological and ultrastructural studies. Liver and kidney tissues were also collected and stored at -200C for estimation of GSH. A significant (P < 0.05) decrease in body weight gains was recorded in group 2. Haematological observations revealed a significant (P < 0.05) decrease in TEC, Hb, PCV, MCV, MCH and MCHC except TLC in group 2. The biochemical assays showed a significant (P < 0.05) increase in serum creatinine, ALT and AST, and decrease in total protein in group 2. The tissue biochemical profile revealed a significant (P < 0.05) decrease in GSH concentration in liver and kidney in group 2. A mild to moderate improvement in all the parameters were observed in group 4 in comparison with group 2 throughout experimental period. Grossly group 2 animals revealed atrophied kidney, abscess and congestion of liver whereas group 4 animals revealed only congestion of liver. Histopathologically, group 2 sections of kidney revealed cystic dilatation of tubules, shrunken glomeruli, vacuolation, presence of haemorrhages and cystic spaces in between tubules. Liver sections showed marked dilation, congestion of central vein, portal vein and sinusoidal spaces. A notable observation was made in hepatocytes like vacuolation/ fatty change and degeneration. Testes revealed vacuolation of semniferous tubules, detachment of germinal cells from basement membrane, increased interstitial spaces, disrupted basement membrane, presence of few leydig cells, severe congestion in interstitial spaces and tunica albuginea. Sections of brain tissue revealed degeneration of purkinje cells, shrunken neurons, vacuolation around neurons, chromatolysis, matrix vacuolation and marked congestion. Group 4 kidney sections showed mild peri glomerular congestion, moderate inter tubular haemorrhages and liver revealed moderate congestion, dilatation of central vein and portal vein and degeneration of hepatocytes. Testes revealed only mild degenerative changes in semniferous tubules whereas brain tissues showed mild congestion and degeneration of purkinje cells. Ultrastructurally, group 2 kidney has evidenced degeneration of tubular epithelium with loose inter cellular junctions, disrupted nucleus, margination of chromatin material (apoptosis), varied size and shape of mitochondria and vacuoles in cytoplasm. Liver section showed swollen nuclei, mitochondrial changes (varied size and shape), disrupted chromatin and rough endoplasmic reticulum. Ultra thin sections of testes showed swollen nuclei, increased perinuclear space, varied size and shape of mitochondria, complete disintegrated chromatin material and degeneration of spermatids. Brain section revealed disruption, margination of chromatin material (apoptotic nuclei) and vacuolar mitochondria. In group 4 animals kidney section revealed dilated inter tubular area, apoptotic nuclei and varied size and shape of mitochondria. Liver section showed swollen nuclei of hepatocytes. Testes section revealed margination of chromatin material, varied size and shape of mitochondria and degeneration of spermatids. Brain section revealed degeneration of neurons. The present study indicated that imidacloprid is a potential toxic agent that induced toxicity at varied levels and resulted in pathological changes in respective target organs viz., in testes, brain, liver and kidney. These changes were well supported by haemotological, serum and tissue biochemical alterations and ultrastructural changes. Vitamin C supplementation provided protective action and moderate improvement in all the above parameters.ThesisItem Open Access TOXICOPATHOLOGY OF λ-CYHALOTHRIN AND ITS AMELIORATION WITH N-ACETYL CYSTEINE IN RATS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2011-12) PALLAVI, G; ANAND KUMAR, A(MAJOR); MADHURI, D; REDDY, M.RABSTRACT : The present experimental study was undertaken to study the pathology of experimentally induced λ-cyhalothrin toxicity in rats. A total of 32 healthy female Sprague dawley rats were 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 8 in each group. The experimental study was designed as follows: Group 1-Control, Group 2-λ-cyhalothrin – toxic control @ 2mg / kg body wt. orally every alternative day for 4 weeks. Group 3- N-acetyl cysteine control @ 300 mg/kg.b.wt per day orally for 4 weeks, Group 4- λ-cyhalothrin @ 2 mg/kg body weight every alternative day for 4 weeks + N-acetyl cysteine control @ 300 mg/kg.b.wt per day orally for 4 weeks. At the end of 2nd week, 2 rats from each group and at the end of 4th week remaining rats were sacrificed; blood and serum were collected for haematological and biochemical studies, tissue samples of liver, kidney, brain, heart and muscle were collected for histopathopathological studies in 10% neutral buffered formalin. Tissues of brain liver, heart and kidney were collected and stored at -200 C for further estimation of oxidative parameters. The effects of λ-cyhalothrin were assessed by studying the various parameters viz., clinical signs, haemato-biochemical, pathomorphological alterations and oxidative stress. During the experiment, rats in group 2 showed reduced feed and water intake, hyperesthesia, mild salivation, incordination of movements. No significant clinical signs were observed in group 3. Haematological studies revealed that overall means of TEC, Hb and PCV were significantly (P<0.05) reduced in group 2 in comparison to other groups. The biochemical assays showed a significant (P<0.05) increase in CPK, LDH, ALT, Serum creatinine and decrease in total protein in group 2. The tissue enzymes assays revealed a significant (P<0.05) increase in TBARS, Protein Carbonyles and significant (P<0.05) decrease of GSH, GST, Na+ K+ ATP ase activities in group 2.The ameliorative group showed mild to moderate improvement in all parameters in comparison to group 2. The gross pathological changes in the group 2 animals included atrophy and necrotic foci on liver, atrophy and medullary congestion of kidney. In group 2, histopathologically liver revealed marked central vein congestion, bile duct hyperplasia, centrilobular necrosis, marked nuclear changes. Kidney revealed marked congestion, disrupted tubular and glomerular architecture, shrunken / atrophied glomeruli marked degenerative changes in tubules with marked intertubular hemorrhages and casts. Brain revealed vacuolation, congestion, neuronal degeneration and chromatolysis. Heart revealed severe disruption, separation, degeneration, necrosis of muscle fibres. Muscle revealed marked disruption, loss of striations and mild hemorrhages. In group 4 animals, liver revealed mild to moderate central vein congestion, dilatation of sinusoidal spaces, degenerative changes in the hepatocytes, kidney showed mild to moderate degenerativ e changes, heart revealed moderate range of degeneration and disruption of muscle bundles. The study indicated that exposure to λ-cyhalothrin result in neurotoxicity and cytotoxicity. Further NAC supplementation provided protective action and improvement in histological, haematological, biochemical and oxidative parameters.ThesisItem Open Access TOXICOPATHOLOGY OF CISPLATIN AND ITS AMELIORATION BY TURMERIC IN RATS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2011-10) RAMYA, B; ANJANEYULU, Y(MAJOR); MADHURI, D; GOPALA REDDY, AABSTRACT: Cancer is one of the most dreadful diseases and currently taking the heaviest toll of human lives, with distant hope of finding an effective and complete cure unless detected and treated in early stages. 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. Cisplatin is one of the most remarkable successes in the war on cancer. The present study was aimed to evaluate the protective effect of turmeric in cisplatin induced toxicity in rats. A total of 48 rats which were divided into 4 groups and treated as follows: Group 1: control, 2: cisplatin control @ 2 mg/kg b.wt, 3: turmeric control @ 0.05 mg/kg b.wt and 4: cisplatin + turmeric each with the above mentioned doses. Body weights were recorded at weekly intervals and organ weights were recorded at the time of sacrifice on day 28. Whole blood was collected for estimation of haematological parameters like RBC, Hb, WBC and PCV. Serum was separated for estimation of biochemical parameters like total protein, BUN, AST, GGT, CPK and creatinine. Studies on whole blood and serum were conducted at fortnightly interval. Visceral organs such as kidney, liver, brain, heart, stomach and intestines were collected for estimation of TBARS, GSH and protein carbonyls in tissue homogenates. Activity of SOD, catalase, G6PD, GPX, GSTNa+-K+ ATPase, Mg2+ATPase and CYP450 was estimated in liver. At the time of sacrifice gross pathological lesions were noted, tissue pieces from kidney, liver, brain, heart, stomach and intestine were collected and preserved in neutral buffered formalin for histo-pahology. Tissue pieces of kidney were also preserved in glutaraldehyde for transmission electron microscopy. Body weight gain, relative organ weight, RBC, WBC, Hb, PCV, serum protein, GSH, SOD, catalase, GST, GPX, G6PD, Na+/K+ ATPase and Mg2+ATPase were significantly (P < 0.05) decreased in cisplatin administered group, while CYP450, TBARS, protein carbonyls, serum BUN, AST, GGT, CPK and creatinine were significantly (P < 0.05) increased in group 2. Group 1 and 3 did not reveal any significant abnormalities. The ameliorative group 4 showed mild to moderate improvement in all parameters in comparison to group 2. Gross pathology of group 2 rats revealed reduction in size of kidney, shrunken liver with small abscess and moderate hemorrhages on lungs. Histopathological sections from kidney of group 2 rats showed marked intertubular hemorrhages, congestion, marked dilation of tubules, shrunken glomeruli and few with disappeared capillary network. Some sections showed hyaline casts, appreciable degenerative changes in tubular epithelium and mild fibrosis. Sections from liver showed marked central vein and sinusoidal congestion, bile duct hyperplasia, few hepatocytes with hydropic degeneration and other degenerative changes. Sections from brain revealed marked congestion of cerebral vessels, mild subcapsular hemorrh-ages, liquefactive necrosis, gliosis and perivascular cuffing. Sections from heart revealed marked interfibrillar congestion, hemorrhages in purkinjee fibres, disrupted cardiac muscle fibres and round cell infiltration. Sections of stomach showed marked congestion and hemorrhages between the villi and also at the base of villi, swollen villi with loss of epithelium at the tips and edematous fluid between villi. Sections from intestine showed marked disruption of villi epithelium. The gross and histopathological lesions in group 4 were mild compared to group 2. Groups 1 and 3 did not reveal any lesions of pathological significance. Ultrastrucural changes in rats of group 2 showed moderate dilation of tubules, varied shapes and sizes of mitochondria, disruption of nuclear chromatin material, perinuclear clumping of mitochondria, marked cytoplasmic vacuolation, margination of chromatin material, disrupted and condensed endoplasmic reticulum, nuclear changes like pyknosis, dilation of nuclear pores and condensation. Rats of group 4 showed nucleus with uniform size, well differentiated nuclear membrane and nucleolus, intact inner and outer nuclear membranes, regenerating mitochondria with prominent inter tubular and inter cellular junctions. From this study, it was concluded that the adverse effects of cisplatin can be reverted by administration of turmeric in a dose-dependent manner.ThesisItem Open Access HAEMATO-BIOCHEMICAL AND PATHOLOGICAL STUDIES OF GUAR MEAL (Cyamopsis tetragonolobn) TOXICITY IN BROILER CHICKEN AND ITS AMELIORATION(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2010-12) DEEPTHI, P; ANJANEYULU, Y(MAJOR); ANAND KUMAR, A; DHANALAKSHMI, KABSTRACT: An experiment was conducted to study the ameliorating effect of DLmethionine (DLM) and polyethylene glycol (PEG) on the guar meal induced toxicity in broiler chicken. During the experimental period six groups of birds (15 in each) were fed with guar meal, PEG, guar meal + DLM, guar meal + PEG, guar meal + DLM + PEG, added to the control diet. Feed and water were offered ad libitium for six weeks of the experimental period. Clinical symptoms and mortality were recorded daily. Data on body weight gains, feed consumption and feed conversion ratio were recorded at weekly intervals. Birds were sacrificed at 2nd, 4th and 6th week of age, to study haematological (PCV, Hb and TEC), biochemical (serum GGT, total proteins and creatinine) parameters and also to study the gross and histopathological changes. In comparison to control (Gr I), body weight gains, feed intake and feed efficiency were significantly (P<0.05), lower in all experimental diets except on PEG diet (Gr Ill), in which the performance parameters were comparable to control(Gr I). Body weight gain, feed intake and feed efficiency were significantly (Pc0.05) improved in ameliorated diets (Gr IV, V and VI) compared to guar meal diet (Gr 11). Inclusion of guar meal (15%) has significantly (P<0.05) decreased the PCV, Hb and TEC compared to control. The toxic effect of guar meal on haematology was significantly (Pc0.05) reduced by DLM supplementation (Gr IV) where as guar meal +PEG (Gr V) did not show much ameliorating effect. The haematological parameters were much more improved in group ameliorated with both DLM and PEG (Gr VI). Significant (Pe0.05) increase in serum GGT levels, creatinine levels and decrease in total proteins were observed in guar meal diet (Gr I!). The group of guar meal with methionine supplementation (Gr IV) had shown a significant ( Pc0.05) decrease in serum GGT levels and significant improvement in serum proteins, but no significant difference was observed in the serum creatinine, compared to the guar meal fed group (Gr 11). The group with PEG supplementation (Gr V) had shown a significant (Pc0.05) decrease in serum GGT and creatinine levels but a significant (Pc0.05) increase in serum total protein levels but no significant difference in serum creatinine levels. The group of guar meal supplementation of both methionine and PEG( Gr VI) had shown a significant decrease in serum GGT and increase in total proteins compared to guar meal fed group (Gr 11) and treatment groups (Gr IV and V), but no significant difference in serum creatinine levels as compared to the guar meal diet (Gr 11). Birds fed with guar meal diet showed moderate gross changes and only mild lesions were observed in the treatment groups. In the guar meal fed group, the lesions include enlarged, pale liver, hypertrophy of pancreas, congested intestine, heart and spleen. Histopathology of liver revealed sinusoidal dilation, bile duct hyperplasia and focal lymphoid aggregates. The intestines revealed much marked disruption of the villi and submucosal congestion. The pancreas revealed moderate congestion in between the acinar cells, kidney showing intertubular congestion and degenerative changes in the tubular epithelium. The heart revealed moderate interfibdllar hemorrhages. The birds belonging to ameliorative groups revealed lesions of only mild significance. Therefore, it can be concluded that DLM and PEG had some amelioration effect and the combination of both DLM and PEG is more effective in ameliorating the guar meal induced toxicity, as evidenced from the results of haematological, biochemical, gross and histopathological changes.ThesisItem Open Access PATHOLOGICAL EFFECTS OF GUAR MEAL AND ITS AMELIORATION BY DETOXIFICATION METHODS AND ACTIVATED CHARCOAL IN BROILER CHICKEN.(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2010-10) RAMU, M; MADHURI, D(MAJOR); ANJANEYULU, Y; GOPALA REDDY, AABSTRACT: The present programme was designed to study the guar meal toxicity and its amelioration in broiler chicken. A total of ninety day-old broiler chicks belonging to vencobb strain were randomly divided into five groups consisting of 18 in each group. Group I was maintained on basal diet, group II on raw guar meal at the rate of 15% in basal diet, group III on raw guar meal + activated charcoal (0.1%) group IV given activated charcoal (0.1%) in basal diet and group V was given alkali treated guar meal at the rate of 15% in basal diet for six weeks respectively. Body weight gain, feed consumption and FCR were recorded in each group at weekly intervals. Five birds from each group were sacrificed at fortnightly intervals and samples for haematological, serobiochemical and histopathological studies were collected. Clinically all the birds were healthy except for a few birds in group II and III which had loose and sticky droppings and pasting of the event. Mortality was not recorded in any of the groups. The group II birds showed significant (P<0.05) reduction in body weight gain and feed consumption and increases in FRC. While group III and V (amelioration groups) showed significant (P<0.05) improvement in weight gain, feed consumption and FCR. Haemtological. Findings revealed that mean values of Hb PCV and TEC significantly (P<0.05) were reduced in group II comparison to all other groups. This might be attributed to disturbances in metabolism that lead to deficiency of clotting factors. The serobiochemical studies showed a significant (P<0.05) reduction in total protein, albumin and globulin levels while significant (P<0.05) increase in the creatinine and AST levels in group II birds. The gross pathological findings observed in guar meal fed group (GII) Heart was apparently normal in all the groups, except for mild congestion and hemorrhages noted in group II birds. Spleen was enlarged and moderately congested while bursa of fabricius was atrophied in group II birds. Moderate to severe congestion and haemorrhages were observed in the intestine of group II birds. The ameliorative groups revealed similar lesions but were mild in nature. The histopathological changes in liver revealed sinusoidal dilatation, congestion of central vein, mononuclear cell infiltration and vacuolar changes in the hepatocytes kidney sections showed congestion, intertubular haemorrhages with degenerative changes in a few tubules. An increase in the interfollicular space and moderate depletion in the follicles were observed in sections of bursa of Fabricius. Spleen sections revealed marked congestion of trabecular arteries. Heart sections revealed marked interfibrillar haemorrhages and heterophilic infiltration. The ameliorative groups (GII and V) showed mild to no pathological lesions in various organs. Among ameliorative groups, lesions were comparatively mild in alkali treated group than activated charcoal treated group. It can be concluded from the present study that guar meal incorporation at the rate of 15% level in feed adversely affected the growth and haematobiochemical parameters of birds. The gross and histopathological studies of various organs also suggested moderate damage to liver, kidney, bursa of Fabricius and intestine. Addition of activated charcoal and detoxification of guar meal by alkali treatment had considerable ameliorating effects on growth, haematobiochemical parameters and microscopic lesions in various organs.ThesisItem Open Access INCIDENCE OF TUMOURS IN POULTRY WITH SPECIAL REFERENCE TO MAREK'S DISEASE AND LYMPHOID LEUCOSlS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2010-10) SWATHI, BORA; ANAND KUMAR, A(MAJOR); MADHURI, D; UDAYA KUMAR, M; REDDY, M.RABSTRACT: The aim of the prescnt study was to investigate thc incidence of' tumours in poultry and diagnose the type of tumor by histopathological and molecular methods. For this purpose, a total of IS9 samples fro111 72 cases were collected and were subjectcd to various diagnostic methods including cytological, rnolccular, histopathological and itn~nu~~ohistochetnicstnuld ies. The incidence of tumours was rccoru .d as highest (46.93'10) in birds ranging from 8 to 18 weeks of age and it was least (4%) in birds of age group less than S weeks. The highest incidence of tumours was recorded in Rajasree breed (46?/0) followed by Rajasree crossbreeds (1 8%), White Leghorns (1 2%). Aseel ( 10Y h), White Leghorn crossbreeds (6%), Vanaraja (6%) and least in Kadaknath brecd (2%). The incidence of tumours was high in female birds (87.5%) when compared with males (12.5%). On the basis of type of tumours, the highest incidence was recorded for Marek's disease (43.05%) followed by lymphoid leucosis (27.77%), other leukotic tumours (8.33%), hepatoma (4.16%), adenoma (4.16%), fibroma (2.77%), haemangioma (2.77%) and least for granulosa cell tumour, nephroblastoma, osteofibroma, endothelioma and chondroma (1.38%). Ninty impression smears of various organs or tissues collected from 46 birds was stained with Leishman's / Giemsa stain. Cytological studies revealed that 67.78% of the smears were diagnosed as Marek's disease, and 23.33% of the smears as l ylnphoid leucosis. Fifty nine tissue samples suspected for MD and LL were subjcctcd for PCR and 50.85% of the samples were positive for MD, produced an amplicon size of 302bp with primers MI and M2. 42.37'/0 of the samples were positive for ALV by using pair of primers (AD1 and H5) and produced an arnplicon of size 326bp. DNA of 6.78% samples were neither amplified for MDV nor for ALV with any of the rcspcctive primers. Grossly, 25%of 72 cases wcre suspected for Marek's disease, 63.SU4 for Lymphoid lcucosis, 9.7Zoh for fibromas and 2.77% for hae~nangiomas. A total of 189 tissue samples frorn 72 cases were subjected to histopathological studies and 120 (63.49%) sainples were confinned as Marck's disease, 55 (39.10%) as LL and 14 (7.4%) as other neoplastic conditions which included cases of adenoma, hepatorna, haenlangioma, fibroma. nephroblastoma. chondroma, granulosa cell tumour, endothelioma and osteofibroma. A total of 29 samples were subjected to immunohistochelllical staining technique for PCNA and observed PCNA labelling in the tumour tissue and demonstrated different levels of staining intensity and indicated the activity of the cells.ThesisItem Open Access PATHOLOGICAL AND MOLECULAR DIAGNOSIS OF MAREK’S DISEASE IN COMMERCIAL LAYER CHICKEN(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2016-11) SRIKANTH, K V; RAMA DEVI, V(MAJOR); ANNAPURNA, P; LAKSHMI KAVITHA, KABSTRACT: Marek’s disease (MD) is a lymphoproliferative disease of chickens caused by Marek’s disease virus (MDV-1). Of late, there are incidences of MD in vaccinated commercial layer chicken in India, causing severe economic losses. Only a few reports are available about the status of the disease in Andhra Pradesh. Hence, keeping this in view the present work was undertaken to study the incidence and gross and histopathological lesions in various organs along with diagnosis of MD by PCR technique. The materials for the present study were procured from Nuzividu, Tanuku and Anaparthy towns, private diagnostic laboratories,Vijayawada and from the post mortems conducted at Department of Veterinary Pathology, NTR College of Veterinary Science, Gannavaram. The duration of the study lasted from December 2015 to October 2016. A total of 520 dead layer birds that were vaccinated previously against the disease were examined in detail and representative samples from suspected MD cases were collected. Of these, 58 birds (11.15%) of 20-52 weeks of age were found positive for MD based on clinical signs, gross and histopathological findings in different organs and by PCR diagnosis. The MD affected chicken exhibited clinical signs of emaciation, weight loss, paleness of comb, marked reduction in egg production, ruffled feathers, diarrhoea, lameness and prostration before death. Out of 58 MD cases, visceral tumors of acute form of MD were noticed frequently in the liver (93.1%), kidney (79.3%), spleen (53.5%) and ovaries (20.7%) and less frequently in the proventriculus (17.2%), intestines (13.8%), skin (10.3%), heart (5.2%) and lungs (5.2%). Grossly, the livers were enlarged and had multiple greyish-yellow tumor nodules of either diffuse, nodular or of miliary form. The kidneys were enlarged, irregular and pale and had pinpoint to a few mm greyish white nodules or masses in many cases. The spleen was enlarged and had miliary greyish white lymphomas. The ovaries were enlarged, irregular, pale or greyish white and revealed firm tumorous mass with cauliflower like appearance. The mucosa of proventriculus and the intestinal wall were thickened and nodular tumors were visible through serosa of intestine. Greyish white feather follicular nodules in the skin, greyish white nodular growths in the myocardium and in the lungs were found. Histologically, in the MD affected organs lymphoproliferative lesions were noticed with infiltration of pleomorphic lymphoid cells and the severity of visceral lymphoid lesions were graded into mild, moderate and severe. The sections of liver in severe cases, revealed diffuse or massive proliferation of pleomorphic lymphoid cells or nodular lymphomas compressing the surrounding parenchyma. In severe cases, diffuse proliferation and infiltration of pleomorphic lymphoid cells, plasma cells and a few MD cells or discrete large lymphomas were noticed compressing the renal parenchyma in the kidneys. In severe cases, spleen revealed proliferation and infiltration of lymphoblasts, small and large lymphocytes, macrophages and reticular cells or multiple or solitary lymphomas in the parenchyma. The lesions in the ovaries were characterized by diffuse pleomorphic lymphoid cell infiltration around the ovarian follicles with distortion of ovarian structure in severe cases. In the proventriculus, diffuse infiltration of pleomorphic lymphoid cells in the mucosa, submucosa and within and in between the glands and in the serosa were evident in severe cases. Marked pleomorphic lymphoid cell proliferation was frequently observed in the mucosa, submucosa, muscle layers and subserosa of intestine and lymphomas with nodular aggregation of large and small lymphocytes were clearly evident in the serosa. The lesions in the skin in moderate cases were characterized by nodular infiltration of pleomorphic lymphoid cells below the epidermis, in the dermis and around the feather follicles. In the myocardium, focal degeneration of muscle fibres, proliferation and infiltration of pleomorphic lymphoid cells and rupture of cardiac muscle fibre bundles were seen. The lung sections showed congestion and marked infiltration of pleomorphic lymphoid cells in the wall of the air capillaries and around the secondary bronchus and parabronchus. The sciatic nerve revealed type A lesions in two birds. In the present study, PCR was carried out for the detection of 132 bp repeats and meq gene of serotype 1 MDV in the tumor tissue samples of MD suspected birds that produced two tandem repeats of 132 bp and the primers specific to meq gene produced an expected size of amplicon in MD positive birds. In the present investigation, the pathological and molecular diagnosis of MD was carried out in commercial layer chicken. It was evident that in spite of vaccination, MD continues to be a major threat causing significant losses in commercial poultry flocks.