Browsing by Author "ANAND KUMAR, A"
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ThesisItem Open Access ADDITION OF A HERB TO A STATIN: A POSITIVE OR NEGATIVE INTERACTION? EXPERIMENTAL STUDIES ON DYSLIPIDAEMIAS IN RATS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2009-03) DILIP REDDY, GUNTURU; GOPALA REDDY, A(MAJOR); SRINIVASA RAO, G; ANAND KUMAR, A; RAJASEKHAR REDDY, AABSTRACT : A total of 56 male Sprague dawley rats of uniform weight and age were randomly divided into seven groups consisting of eight rats in each group after an acclimatization period of 3 weeks to evaluate the interaction of atorvastatin with garlic in induced dyslipidaemia. Group 1 served as plain control, while groups 2 and 3 were fed with high fat and high cholesterol diet throughout the experimental period. Groups 4,5,6 and 7 received 1% (100% dose), 0.5% (50% dose), 0.25% (25% dose) and 0.75% (75% dose) fresh garlic w/w in feed, respectively in addition to the high fat and high cholesterol diet and administered with 10 (100% dose), 5 (100% dose), 7.5 (100% dose) and 2.5 (25% dose) mg/kg atorvastatin respectively, while group 3 served as atorvastatin control, which received 10 mg/kg atorvastatin per day orally for 12 weeks. Blood collection was carried out at every two weeks interval for plasma biochemical analysis of total cholesterol, HDL cholesterol, triglycerides and creatinine and aspartate transaminase (AST). Single dose and multiple dose pharmacokinetic studies were performed at the beginning of the first dose and at the end of last dose of atorvastatin, respectively in groups 3 to 7. At the end of the experiment, liver and kidneys were collected for assay of TBARS, glutathione and SOD. Histological, histochemistry and electron microscopy studies were conducted on different organs at the end. All the treatment groups exhibited significant improvement in dyslipidaemic condition when compared with group 2 from 2nd week of treatment by reducing the TC, TG and LDL-C levels with subsequent increase in HDL-C levels. Group 4 was highly effective in correcting dyslipidaemia due to the synergistic pharmacodynamic actions of herb and drug. Plasma atorvastatin concentrations during multiple dose PK studies were significantly higher than single dose counterparts. PK parameters showed a significant increase in the garlic treated groups with high values of Cmax, AUC, AUMC, MRT and half-life which could be attributed to the inhibitory activity of garlic on drug transporters and metabolizing enzymes. High concentration of the drug in plasma in group 4, 5 and 3 resulted in toxicological manifestations in liver and kidney, which was evident from the increased plasma creatinine concentration, AST activity and oxidative stress. Histopathological studies on liver, kidney revealed moderate to severe damage in groups 4 and 5, which also exhibited mitochondrial damage on transmission electron microscopy. From this study, it can be concluded that garlic and atorvastatin exhibited positive pharmacodynamic interaction in reducing dyslipidaemias. The pharmacokinetic studies revealed that garlic increased the pharmacokinetic parameters and the toxicological studies indicated that high dose of atorvastatin + garlic has negative safety profile. Further studies are warranted to address the pharmacokinetic interactions of statin and garlic in detail.ThesisItem Open Access ARSENIC (As) INDUCED TOXICITY AND ITS AMELIORATION IN BROILER CHICKEN(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2007-10) PADMAJA, BHYRU; MADHURI, D(MAJOR); ANAND KUMAR, A; DHANA LAKSHMI, KABSTRACT: Arsenic is a heavy metal available abundantly in the air as an industrial pollutant and resulting oxidative stress in livestock as well as in poultry. The present study was designed to study the arsenic induced toxicity and its . amelioration in broiler chicken. A total of 75 sexed male broiler chicks of day old age belonging to Vencobb strain were randomly divided into five groups consisting of fifteen chicks in each group. Group I was maintained on basal diet and group II on arsenic @ 100 ppm in feed for 6 weeks. Group Ill was maintained on Emblica officinalis @ 500 ppm for 6 weeks. Group IV was given with arsenic @ 100 ppm along with Emblica officinalis @ 500 ppm for 6 weeks. Group V was given arsenic @ 100 ppm containing diet for the first 4 weeks and subsequently treated with Emblica officmalis @ 500 ppm for the remaining 2 weeks. Individual 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. Blood, serum and tissue samples were collected. Biomarkers of hepatic damage, protein profile in serum, TBARS, GSH in liver and kidney were estimated. The histopathological studies of various vital organs were carried out at the end of 2nd4, tha nd 6" week. HI titre in serum and PHA index were estimated at the end of 4" and 6" week to evaluate the immune status of birds. The arsenic treated group resulted in significant (Pe0.05) reduction in body weight gain, feed consumption and increase in FCR. Haematological studies revealed that overall mean values of PCV, Hb and TEC were significantly (P<0.05) reduced in group II in comparison to other groups. The biochemical assays showed significant (Pe0.05) reduction in total protein, albumin, globulin while significant (P<0.05) increase in A/G ratio, GGT, Creatinine and BUN. Studies on oxidative stress revealed significant (Pc0.05) increase of TBARS (Liver and Kidney) and significant (Pc0.05) decrease of GSH (Liver and Kidney) in the group II. lmmunological assays in group II revealed significant (P<0.05) increase in HI titre and significant (Pc0.05) reduction in PHA index in comparison to other groups (1, Ill, IV and V). Incorporating herb in the treatment groups (IV and V) showed marked improvement in all the above parameters in comparison to arsenic toxic control group. Clinically, birds in group II and V from 3d week onwards showed reduced feed consumption, weight gain while the birds in other groups were normal. Grossly, mild haemorrhages with rounded borders and pale discoloration of liver with mild swelling and haemorrbages in kidney was noticed. Reduction in size and mild congestion was observed in bursa of Fabncius and spleen. Few haemorrhages with mild congestion of heart in the earlier stages and later similar lesions with increased severity were obsewed. Group V also revealed similar types of lesions upto 4 weeks and mild lesions at later stages. No lesions of pathological significance were observed in other groups. The histopathological changes in liver revealed dilatation of sinusoidal spaces, congestion and bileduct hyperplasia with infiltration of mononuclear cells. Kidney sections showed mild to moderate intertubular haemorrhages with degenerative changes in few tubules. Mild to moderate depletion of lymphocytes with interfollicular fibrosis and haemorrhages was observed in sections of bursa of Fabricus. Spleen sections showed mild depletion of germinal centers with marked congestion of trabecular arteries. Mild disruption of cardiac fibres with moderate interfibrillar haemorrhages was observed in heart sections. The treatment groups (IV and V) revealed similar lesions but are of mild in nature. It can be concluded that arsenic @ 100 ppm in feed for 6 weeks caused a significant toxicity in broiler chicken and addition of antioxidant Emblica officinalis was efficient in ameliorating the oxidative damage induced by arsenic.ThesisItem Open Access COGNITIVE AND NEURO-ENDOCRINE DISRUPTION OF LEAD AND MONOCROTOPHOS AND THEIR RELATION TO THYROTOXICITY IN PERINATALLY EXPOSED RATS(SRI VENKATESWARA VETERINARY UNIVERSITY , TIRUPATI – 517502. (A.P.) INDIA, 2010-06) KALA KUMAR, B. D. P; GOPALA REDDY, A (Major); RAVI KUMAR, P; KONDAL REDDY, K; ANAND KUMAR, AABSTRACT : Thyroid hormone is essential for neuronal and glial genesis and also the time specific migration of neurons. Any change in the sequential neurodevelopment of foetus or the neonate would be manifested as behavioural abnormality in the adult life. In utero exposure to xenobiotics would interfere with the availability of maternal thyroid hormone to the foetus. Pesticides and heavy metals form a major chunk of the environmental pollutants that affect the behaviour of animals and human beings. Monocrotophos a widely used pesticide and lead a ubiquitous heavy metal are known neurotoxicants. The role of these two substances in thyroid disruption and subsequent developmental neurotoxicity was studied. Thirty pregnant female rats were divided into five groups. Group I was Sham. Methimazole (II), monocrotophos (III), lead acetate (IV) were administered singly and in combination (V) to assess the interaction. AChE, thyroid profile (TSH, T3 and T4), maternal behaviour, litter size, neonatal mortality, neurodevelopmental (brain wet weights, DNA, RNA and protein), neurobehavioural (auditory startle response, rope descent, mid air righting reflex, elevated plus maze, photoactometry and morris water maze) and neurochemical (acetyl choline and glutamate content of the brain) parameters were studied. Histopathology of thyroid and brain were conducted. Inhibition of AChE was < 20% in III and V. Thyroid profile decreased in II and T4 increased in IV. Maternal behaviour was significantly (p<0.01) interfered in III and V. Neurodevelopmental and neurobehavioural parameters did not reveal significant changes. Glutamate content was highest in group V indicating excitotoxicity. Thyroid was affected significantly in II, III and IV but not in V. Cerebral cortical layers were affected in groups II through V. The three layers of cerebellum either had abnormal arrangement or decreased cellularity in all treated groups. Thus, it is concluded that monocrotophos and lead acetate could act as thyroid disruptors and might have interfered with neurodevelopment during the perinatal exposure. Group V also affected neurodevelopment but did not affect thyroid histology suggesting other mechanisms could have contributed to the neurotoxicity.ThesisItem Open Access DEVELOPMENT AND STANDARDIZATION OF RAPID DIAGNOSTIC ASSAYS FOR PPR VIRUS DETECTION(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2007-01) KEERTI MEENA; JANAKIRAMA SARMA, B(MAJOR); NARASIMHA REDDY, Y; ANAND KUMAR, AABSTRACT : Peste des petits ruminants (PPR) is a highly contagious viral disease of sheep and goats resulting in heavy mortality and morbidity and is considered to be one of the main constraints to improving productivity of small ruminants in the regions where it is endemic. Sever economic losses are reported annually from different parts of India due to absence of timely detection of the disease. Rapid and accurate detection of PPR is important for efficient control and epidemiological surveillance and to reduce losses to the livestock industry. Therefore, there is a need for the development of rapid diagnostic tests in order to have a better control over the disease. The present study was taken up with an aim to standardize and develop rapid diagnostic methods such as Latex agglutination test (LAT), capture ELISA and RT-PCR for the detection of PPR virus in infected cell culture fluid and clinical samples. The tests were standardized using PPR virus infected and uninfected cell culture (as control) fluids. The test were then used to detect PPR virus/ antigen in 34 clinical samples comprising of 4 oral, 4 nasal, 2 lachrymal and 2 fecal swabs and 22 tissue samples. PPR virus (vaccine strain) from infected cell culture fluid was purified and concentrated using PEG 6000. The concentrated virus was used for raising hyperimmune serum in rabbits. The levels of antibodies in the serum were monitored by AGID. Immunoglobulins were purified from serum using saturated ammonium sulphate. LAT was standardized using purified antiPPR rabbit Ig tagged to latex beads. Both coloured (blue) and white beads were used. The test was found to be simple and rapid. The shelf life of LAT reagents at room temperature and 4oC was also studied. The reagent was found to be stable for 2 wks. at room temperature and at 4oC for 7 wks. (maximum period tested). Thirty four clinical samples were screened by LAT, of which 20 (58.82%) were found to be positive for PPR. Capture ELISA was standardized employing reagents supplied by I.V.R.I and it identified 21 positives with a per cent positivity of 61.76. RT-PCR was standardized and nested PCR was done in order to confirm the first stage PCR product which yielded 309 bp products. Out of total 34 samples 18 samples were positive for presence of PPR virus by RT-PCR. None of the nasal, fecal and eye swabs were positive for amplification product by RT-PCR. Sensitivity and specificity of LAT was found to be 90.4 % and 92.3 % respectively when capture ELISA was reference method. When RT-PCR was reference method, LAT showed sensitivity of 88.8% and specificity of 75%. It may be concluded that in the present study LAT was found to be equally sensitive and specific to capture ELISA but inferior to RT-PCR. By using monoclonal antibody as in capture ELISA, the sensitivity and specificity of LAT could be improved further to make it an ideal field test for the rapid diagnosis of PPR in outbreak areas and to plan effective strategies to control disease.ThesisItem Open Access DIAGNOSIS OF BOVINE SARCOCYSTOSIS BY IMMUNOFLUORESCENT ANTIBODY TECHNIQUE(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2012-03) DASMA BAI, BANOTHU; UDAYA KUMAR, M(MAJOR); CHENGALVA RAYULU, V; NARASIMHA REDDY, Y; ANAND KUMAR, AABSTRACT: Sarcocystis species are widely prevalent in man and animals, causing significant impact on animal and public health throughout the world. A laboratory standardized immunofluorescent antibody technique was used to study the seroprevalence of bovine sarcocystosis and the efficacy of same was compared with the traditional diagnostic methods like macroscopic, microscopic (squash and pepsin HCl acid muscle digestion). Histopathological changes and viability of bradyzoites in the affected esophageal muscles were also studied in the present investigation. The gross examination of oesophagi revealed creamy white colored thin walled macrocysts of Sarcocystis spp appearing in different shapes (fusiform, elliptical, ovoidal and globular etc) and sizes ranging from 2.0-18.0 x 1.0-5.0 mm with an average size of 10.47 + 0.295 x 3.08 + 0.089 mm. None of the organs showed any kind of gross lesions around the macrocysts embedded in the muscles. Microscopic examination of esophagus and diaphragmatic muscles by squash technique revealed the presence of microcysts arranged horizontally in between the muscle fibers of esophagus where as the pepsin HCl acid digestion of muscle samples of esophagi and diaphragm showed live bradyzoites in gliding motion. Histopathological studies suggested two possible etiologic agents of bovine sarcocystosis namely: S. cruzi, characterized by having elongate and septate cysts and S. hirsuta or S. hominis, characterized by having spherical or rounded cysts with thick radially striated cyst wall. The muscle degeneration with focal or diffused mononuclear cells viz: leukocytic infiltration, eosinophils, lymphocytes and macrophages observed in the tissues under study were attributed to the pathogenic effects of S. cruzi. The ruptured or degenerated state of some of the mature sarcocysts surrounded by eosinophils indicated the advanced age of the cyst. Immunofluorescent antibody technique was standardized in the laboratory for the diagnosis of Sarcocyst infection in bovines. Purified, host cell free bradyzoites collected from macrocysts of Sarcocystis spp, in aliquots of 6-8 applied to glass slides and fixed in chilled acetone over night followed by preservation at -200C worked well with 1:16 dilution of positive and negative control sera and 1:40 dilution of rabbit anti-bovine FITC conjugate. The positive sera did not show any cross reaction with T. gondii RH strain and non specific reactions were absent with negative sera. The serosurveillance of bovine sarcocystosis by laboratory standardized IFAT showed 80.14% (323) of cattle and 78.59% (246) of buffaloes positive for anti sarcocystis antibodies out of 403 cattle and 313 buffalo sera tested, respectively showing an overall prevalence of 79.46% out of 716 animals screened. The antibody titers of 6 randomly selected positive samples from different age groups of <2 years, 2-5 years, 5-10 years and >10 years old bovines ranged from 16-64, 32-256, 32-128 and 16-64 with an average titer of 32 + 2.92, 106.6 + 34, 74.6 + 17 and 34.6 + 9, respectively. The age wise prevalence of sarcocystosis in cattle indicated low rate of infection in the age group below 2 years (60%) and an ascending rate of infection in the age groups of 2-5 years (81.33%), 5-10 years (80.52%) and above 10 years (90.9%). Similarly, the incidence was significantly low in the buffaloes of below 2 years (64%) and high percentage of infection (86.51%) in 5-10 years followed by 78.94% in 2-5 years and 77.27% in above 10 years of age groups. No significant difference of infection was observed between male (81.87%) and female (75.19%) animals as well as between non-descriptive (79.63%) and cross bred (77.58%) animals. Esophageal and diaphragmatic muscle samples collected from 100 animals slaughtered at Chengicherla slaughter house, Hyderabad were subjected to visual examination, squash and pepsin HCl acid muscle digestion techniques which revealed the presence of macrocysts in 16% and 0%, microscopic sarcocysts in 8% and 0% and bradyzoites in 76% and 52% esophageal and diaphragmatic muscles, respectively. The sera collected simultaneously from corresponding animals were screened for anti Sarcocystis antibodies by laboratory standardized IFAT and the results were compared with those of visual examination, squash and pepsin HCl acid muscle digestion techniques. The IFAT was found superior in diagnosing sarcocystosis with positivity of 82%, followed by muscle digestion, gross examination and squash techniques with positive rates of 52%, 16% and 8%, respectively. The present study indicated that the visual and microscopic examination of bovine carcass is by no means a satisfactory diagnostic tool and recommends Immunoflourescent antibody technique for the antemortem diagnosis of animals waiting for slaughter at abattoirs in large scale. Experiments were also undertaken to determine whether Sarcocystis would survive storage at different refrigeration temperatures for a period of 9 days. The number of live and dead bradyzoites in one gram of pepsin HCl acid digested bovine esophageal muscle samples previously stored at room temperature, 40C, 00C, and -200C for a period of 48thhr, 8 days, 24thhr, and 24hr were 2x104 and 4 x104, 1x104 and 1 x104, nil and no bradyzoite, respectively when compared to those stored at 0th hr (10x104 and 0 x104).ThesisItem Open Access Effect of synbiotic (L.casei strain 17 & Fructo- oligosaccharides) on induced colon cancer in rats(SRI VENKATESWARA VETERINARY UNIVERSITY , TIRUPATI – 517502. (A.P.) INDIA, 2013-03) VIKRAM KUMAR, B; KALAKUMAR, B (Major); GOPALA REDDY, A; KONDAL REDDY, K; ANAND KUMAR, AABSTRACT : Microorganisms that favour or help other living beings are called probiotics. Synthesis or fusion of such probiotics with complex carbohydrates such as fructo oligosaccharides is known as synbiotics. These synbiotics are known to mollify the changes that occur in inflammation. The effect of L.casei strain 17 and fructo oligosaccharide on chemical induced colon cancer was studied in Sprague Dawely female rats. A total of 32 rats were divided into 4 groups and treated as follows: Group I- Sham control given normal saline S.C every week for 6 weeks, Group II- DMH control 1,2 dimethyl hydrazine @40mg weekly for 6 weeks S.C. Group III- 1,2 dimethyl hydrazine @40mg weekly for 6 weeks S.C. followed by prebiotic 0.02gm/day and probiotic 109-1011 CFU/day coated with 1% sodium alginate Group IV: 1,2 dimethyl hydrazine @40mg weekly for 6 weeks S.C. along with uncoated synbiotic (Prebiotic-0.02gm/day and Probiotic 109-1011 CFU/day mixed in distilled water) Average body weights were recorded at weekly intervals and blood collected after sacrifice for haematology and sero-biochemical parameters (total cholesterol, calcium, glucose, proteins). Liver and colons were collected for estimating TBARS, SOD, total proteins, GSH, protein carbonyls and colons were also used for estimation of aberrant crypt foci. Colonocytes were isolated and measured to assess the extent of DNA damage by comet assay. Aberrant crypt foci (ACF) were increased significantly (p<0.01) in DMH induced group and uncoated synbiotic group, where as in coated synbiotic group they were decreased. In DMH control and uncoated synbiotic group, the comet appeared to be significantly longer in length compared with coated synbiotic group. Haematological parameters were not allied significantly except for certain changes in total platelets, MCV and lymphocytes. Cell architecture and nuclear cytoplasmic ratio were decreased in coated synbiotic treated group in comparison with DMH control and uncoated synbiotic treated groups. Anti oxidants (GSH and SOD) were significantly (p<0.05) reduced in DMH control and uncoated synbiotic treated groups, where as there was restoration in coated synbiotic group, TBARS and protein carboryls were reduced in DMH and uncoated synbiotic treated groups, while they were restored in coated synbiotic group as compared with control group. Thus, it is concluded that coated synbiotics are more effective as antioxidants in preventing and countering oxidative stress by facilitating restoration of antioxidant defenses as well as decreasing DNA damage in colorectal cells. Hence, their supplementation would reduce various stages of colon cancer.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 INTERACTION STUDIES ON GYMNEMA SYLVESTRE WITH GLIMEPIRIDE AND INSULIN IN EXPERIMENTAL DIABETES MELLITUS IN RATS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2013-12) Srikanth, M.K; GOPALA REDDY, A(MAJOR); BHARAVI, K; MADHAVA RAO, T; KONDAL REDDY, K; ANAND KUMAR, AABSTRACT: An experimental study was conducted to evaluate the interaction of Gymnema sylvestre extract with insulin and glimepiride in diabetic Sprague dawley rats. Rats were randomly divided into 7 groups of 6 rats in each and blood glucose was estimated to ascertain group differences, if any. Group 1 was kept as normal control. Remaining 6 groups were induced diabetes by intraperitoneal injection of streptozotocin @ 40 mg/kg body weight. After 72 h, rats with blood glucose value of >200 mg/dl were included in the study (n=6). Treatment protocols were initiated 48 hrs post-confirmation of diabetes and continued for 2 months. Group 1: non-diabetic control, group 2: streptozotocin (40 mg/Kg i/p single dose)-induced diabetic (DM) control, group 3: Insulin treatment (4 U/kg b. wt. subcutaneously once daily), group 4: glimepiride treatment (4 mg/kg b. wt. orally once daily), group 5: Gymnema sylvestre methanolic leaf extract treatment ( 400 mg/kg b.wt. orally once daily), group 6: Insulin + Gymnema sylvestre methanolic leaf extract treatment (once daily) and group 7: glimepiride + Gymnema sylvestre methanolic leaf extract treatment (once daily). Blood glucose, body weights, sero-biochemical parameters, antioxidant profile in liver, kidney, brain and testis, ATPases, glucose 6 phosphate dehydrogenase (G6PD), cytochrome P450 (CYP450) activity and glycogen in liver, electron microscopy and histopathology of various tissues were studied at different time intervals. Also, pharmacokinetic interaction of glimepiride with Gymnema sylvestre extract was assessed. There were significant alterations in blood glucose, body weights and other biochemical parameters in diabetic control group 2 as compared to group 1. All the treated groups revealed significant improvement in all the parameters as compared to group 2, while the combination treatment in groups 6 and 7 was found better as compared to single agent-treated groups 3, 4 and 5. The histological studies revealed marked changes in group 2 in all the organs studied, while groups 3 to 5 revealed moderate changes and groups 6 and 7 revealed either minor changes or no pathologically significant changes. Group 1 was devoid of any histological alterations. The electron microscopy of kidney, pancreas and aorta revealed marked alterations in group 2, while groups 6 and 7 revealed better architecture. The pharmacokinetic study revealed the values of T1/2 (h), Ka (h-1), Ke (h-1) and Tmax (h) of glimepiride were siginificantly varied in Gymnema sylevestre pre-treated rats compared to normal rats administered with glimperide In conclusion, the study revealed that addition of Gymnema sylvestre leaf extract to insulin and glimepiride had positive pharmacodynamic interaction in improving the patho-biochemical alterations due to streptozotocin-induced diabetes mellitus in rats, which was evident from greater improvement in sero-biochemical and organ parameters in the groups that were treated using a combination of Gymnema sylvestre with either insulin or glimepiride as compared to individual agent-treated groups. Important pharmacokinetic parameters did not vary significantly when glimepiride was used in combination with Gymnema sylvestre leaf extract.ThesisItem Open Access ISOLATION AND CHARACTERIZATION OF AVIAN LEUKOSIS VIRUS FROM BREEDER FLOCKS OF CHICKEN(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2010-10) GOPALA, LUNAVAT; NARASIMHA REDDY, Y(MAJOR); DHANA LAKSHMI, K; ANAND KUMAR, A; REDDY, M.RABSTRACT: The present study was takcn up with a view to isolate a\.ian Icukc,sis virus (ALV) from aft'ected hrecdcr flocks of chickens and characterize the isolatc(s)with rcgard ttr group specitic ac-ELISA, growth in cell culturc titration. serum neutralization, polymerase chain reaction multiplc scqucnce alignment and phylogenetic allalysis in diagnosis of avian leucc~sisv irus infection. 276 cloacal swab sarnples wcrc collected fi-on1 hrccder Ilocks 01' chicken suspcctccl li>r avian lcukosis viral inl'ections. The breedcr flocks of chickcn cxhib~trd sympto~nsli ke tumours in livcr, splccn and heart. Thc sa~nplcs\s Jcr-ct cstcd tbr ALV by group spccilic antigen capture El-ISA ;is a prcliniinnry test hcliirc attcnipts t o isolate the virus. A total of47 sa~nplesw crc positive of'270 samples by cn~pioying p27 ac-ELISA kit. DNA from 16 blood samples (buffy coat) and RNA from 25 cloacal swabs obtained from ALV gs antigen positive flocks were tested for ALV specific sequences by PCR Attempts were made to isolate avian leukosis virus from these cloacal swab samples by passaging in CEF cells. The samples were passaged five times in cell lines. The presence of virus was demonstrated at different passage levels by ac-ELISA and Polymerase chain reaction (PCR). The RT PCR using H5 and AD1 was found negative for the SVVU-I01 isolate where as RT-PCR using primers H5 and H7b was positive with expected product size of 544 bp, which indicate that SVVU-I01 belongs to ALV subgroup-.I. Virus neutralization results indicate that the homologous antiserum efficiently neutralized ALV (SVVU-I 01) isolated in this study Thc nucleotide sequence of gp85 and gp37 was determined for tht: field isolate SVVU- 10 I and compared with publishcd sequences of' ALV subgroups A. B. C. D, E and J and seven strains of' ALV subgroup-.I. The result of prrsent study showed that the SVVU- I0 I belongs to ALV subgroup-J.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 PHARMACOLOGICAL EVALUATION OF HERBAL METHIONINE IN METHIONINE DEFICIENCY AND IRON INDUCED STRESS IN BROILERS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2009-12) SAI GOPAL, T; USHA RANI, M(MAJOR); GOPALA REDDY, A; ANAND KUMAR, AABSTRACT: A total of 120 sexed male broiler chicks of Vencobb strain of day-old age were randomly divided into 8 groups consisting of fifteen chicks in each group. Group 1 was maintained on methionine deficient diet and groups 3, 5 and 7 were supplemented with herbal methionine at level 1 and 2, and synthetic methionine, respectively. Group 2 was maintained as iron added methionine deficient diet and groups 4, 6 and 8 were supplemented with herbal methionine at level 1 and 2, and synthetic methionine, respectively. All the groups were maintained on iso-nitrogenous and iso-caloric diet for a period of 6 weeks. The performance parameters were recorded at weekly intervals. Antioxidant defense profile, biomarkers of hepatic damage, renal damage, protein profile and lipid profile were carried out at 2"d, 4'h and 6th week. At 5m week phytohaemagglutinin (PHA) index and at the end of 6th week histopathological studies were carried out. The methionine deficient and iron added methionine deficient diet groups had a significant (Pe0.05) reduction in body weight, GSH, activity of SOD and catalase, and PHA index, while FCR, and the concentration of TBARS, protein carbonyls and serum creatinine, and the activity of AST were significantly (Pc0.05) increased. Supplementation with herbal methionine at level 1 and 2 respectively in groups 3 and 5 resulted in a marked improvement in all the above parameters as compared to those of methionine deficient diet. Supplementation of herbal methionine at level 2 revealed the performance comparable with synthetic methionine supplementation. Histological abnormalities were also recorded in the liver, kidney, spleen and bursa in all groups, while the groups, 5 and 7 did not reveal any abnormalities on histopathology, while the treated groups 3, 5 and 7 revealed lesions of mild intensity or signs of regeneration. Thus, it is concluded that deficiency of methionine alone, and iron also induces biological damage by means of oxidative stress and the herbal methionine in test offered better performance. The beneficial effects of herbal methionine may be attributed to its antioxidant, anti-stress, hepato-protective principles and biological utilization was as good as synthetic methionine.ThesisItem Open Access PHARMACOLOGICAL EVALUATION OF HERBAL NEONATAL CHICK CARE IN BROILERS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2009-11) CHANDRAVATHY, JADA; GOPALA REDDY, A(MAJOR); USHA RANI, M; ANAND KUMAR, AABSTRACT : A total of 130 sexed male broiler chicks, immediately after hatch, belonging to Vencobb strain were randomly divided into six groups consisting of twenty five chicks each in groups 1, 2, 3 and 4, and fifteen each in groups 5 and 6. Group 1 was maintained on basal diet, group 2 on herbal neonatal chick care @ 6g/chick/day for 2 days after hatching and later continued with basal diet up to 42nd day (6 wks). Group 3 was kept on herbal neonatal chick care @ 8g/chick/day for 2 days after hatching and later continued with basal diet up to 42nd day (6 wks). Group 4 was given FeSO4 @ 0.5% of feed for 42 days (6 wks). Group 5 was given herbal neonatal chick care @ 6g/chick/day for 2 days after hatching and later continued with the FeSO4 @ 0.5% of feed up to 42nd day (6 wks). Group 6 was given herbal neonatal chick care @ 8g/chick/day for 2 days after hatching and later continued with the FeSO4 @ 0.5% of feed up to 42nd day (6 wks). The performance parameters were recorded at weekly intervals. Antioxidant defense profile, biomarkers of hepatic damage, renal damage, lipid profile, protein profile and HI titre in serum were estimated at the end of 4th and 6th wk. Histopathology and estimation of TBARS, GSH, protein carbonyls, HI and phytohaemagglutinin (PHA) index were done at the end of 6th wk. The ferrous sulphate treatment in group 4 resulted in significant (P<0.05) reduction in body weights, protein profile, GSH (6th week), HDL cholesterol, HI titre and PHA index (6th week), while FCR, total cholesterol, LDL cholesterol, triglycerides, TBARS (6th week), ALT, CPK and creatinine were significantly (P<0.05) increased at the end of 4th week and a similar trend was continued at the end of 6th week. Treatment with herbal neonatal chick care in groups 2, 3, 5 and 6 resulted in a marked improvement in all the above parameters as compared to those of ferrous sulphate toxic control group 4 at the end of 6th week. Histological abnormalities were also recorded in the liver, kidney and other tissues in ferrous sulphate toxic control group 4. Groups 1, 2 and 3 did not reveal any abnormalities on histopathology, while the treated groups revealed lesions of mild intensity or signs of regeneration. Thus, it is concluded that ferrous sulphate induces biological damage by means of oxidative stress and the herbal neonatal chick care offered protection and proved beneficial in resisting the adverse effects of stressorThesisItem Open Access PROTECTIVE EFFECT OF N-ACETYL CYSTEINE AGAINST ARSENIC-INDUCED TOXICITY IN RATS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2010-06) HEMALATHA, P; GOPALA REDDY, A(MAJOR); USHA RANI, M; ANAND KUMAR, A; RAMANA REDDY, YABSTRACT: N-acetyl cysteine was evaluated against arsenic-induced toxicity in rats. The Wistar rats were divided into 4 groups and treated as follows: Group 1: sham control, 2: arsenic control, 3: N-Acetyl cysteine (NAC) pre-treatment for two weeks followed by arsenic + NAC and 4: arsenic + NAC. Average body weights were recorded at weekly intervals and testes weights were recorded at the time of sacrifice. On 29th day, organs were collected for estimation of TBARS, protein carbonyls and GSH in tissue homogenates. Activity of Na+-K+ ATPase , Mg2+ATPase and CYP450 of liver, intra-testicular LDH, serum creatinine, and serum LDH were also estimated. Histopathology of heart, liver, kidney, testis, lung, intestine and stomach was also studied at the end. Body weight gain, relative testis weight, GSH, CYP450, Na+/K+ ATPase and Mg2+ATPase were significantly (P < 0.05) decreased, while TBARS, protein carbonyls, serum LDH, intra-testicular LDH and serum creatinine were significantly (P < 0.05) increased in group 2 as compared to other groups. Group 1 did not reveal any abnormalities on histopathology. Group 2 (arsenic control) showed marked degenerative changes in heart, kidney, liver, testis, lung, intestine and stomach. NAC-treated groups (3 and 4) showed improvement in all the parameters studied, though it was marked with NAC pre-treatment. From this study, it is concluded that arsenic induces toxicity to heart, kidney, liver, testis, lung, intestine and stomach, and these effects can be reverted by NAC administration.ThesisItem Open Access SAFETY AND IMMUNOGENICITY OF BRUCELLA ABORTUS STRAIN-19 REDUCED DOSE VACCINE THROUGH CONJUNCTIVAL ROUTE IN CATTLE - A TRIAL(SRI VENKATESWARA VETERINARY UNIVERSITY , TIRUPATI – 517502. (A.P.) INDIA, 2013-04) MANOJ KUMAR, V; NARASIMHA REDDY, Y(Major); DHANALAKSHMI, K; ANAND KUMAR, AABSTRACT : The present study is taken up with a view to study the efficacy of brucella vaccine at reduced dose (5 x 109 to 8 x 109 CFU/dose) through conjunctival route and compare it with conventional standard dose (40 x 109 to 80 x 109) vaccine. Female cattle above 8 months age were selected for the study. Thirty animals were inoculated with reduced dose conjunctival vaccine and ten with standard subcutaneous vaccine using Brucella abortus S19 antigen. The efficacy of vaccine was studied in terms of both humoral and cell mediated immune responses against brucella for a period of 120 days. Humoral immunity techniques consisted of rose bengal plate test (RBPT), standard tube agglutination test (SAT) and indirect ELISA (i-ELISA). Cell mediated immune response was studied by using interferon gamma (IFN-γ) assay after in vitro stimulation of lymphocytes with both S19 and B. abortus 544 as antigens in indirect competitive ELISA (Enzyme Linked Immunosorbent Assay). Of the humoral immunity tests employed for studying the efficacy of vaccines RBPT was least sensitive. It detected 20 to 18.51% positive responders at 21 and 60 days PV in reduced dose conjunctival vaccine group. Similarly it detected 60% and 50% responders on 21 and 60 days PV in standard group. Standard tube agglutination test (SAT) detected more responders against vaccinates than the RBPT. The positive percentage ranged from 26 to 66% among reduced dose vaccine group and 40 to 80% among standard dose vaccine group. I-ELISA was the most sensitive as it detected up to 78.9% responders among reduced dose vaccination group and up to 100% in standard dose vaccinates group. Humoral immune responses declined after 90 days PV. IFN-γ production as indicator of cell mediated immune response in vaccinates was more consistent. IFN-gamma production was observed during the entire period of the trial (120 days). It is concluded that CMI responses are better indicators of immune response to brucella than the humoral immunity.ThesisItem Open Access TOXICODYNAMIC INTERACTION OF LEAD WITH CADMIUM AND THERAPEUTIC EVALUATION OF N-ACETYL L-CYSTEINE IN RATS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2011-07) ANlL KUMAR, B; GOPALA REDDY, A(MAJOR); RAVl KUMAR, P; MADHAVA RAO, T; ANAND KUMAR, AABSTRACT: An experimental study was conducted to evaluate the molecular mechanisms of lead and cadmium toxicity and their toxicodynamic interaction, and to evaluate therapeutic potential of N-Acetyl L-cysteine (NAC) against the toxicity in Wstar rats. After an acclimatization period of 2 weeks, rats were randomly divided into 8 groups comprising of 6 rats in each. Group 1 was kept as normal control throughout the experimental period, 2 was given NAC @ 300 mg per kg body weight administered by oral gavage, 3 was given lead (lead acetate @ I000 ppm in feed), 4 was given cadmium (cadmium chloride @ 300 ppm in feed), 5 was given lead + cadmium as per above doses in feed, 6 was given lead + NAC as per above schedule, 7 was given cadmium + NAC as per above schedule, and group 8 was given lead + cadmium + NAC as per above schedule for 3 months. Body weights, haematology (TEC, TLC, Hb, PCV, MCH and MCHC), activity of 6-ALAD and erythrocytic SOD, sero-biochemical parameters (ALT, CPK, troponins, plasma TBARS and serum creatinine), antioxidant profile (GSH, GST, TBARS and protein carbonyls) in liver, kidney, heart, testis and brain, ATPases and tissue lipids in liver and brain, neurotransmitters (Ach and glutamate) in brain, CYP450, glycogen and G6PD in liver, weight of testes, testicular LDH and sperm count, electron microscopy of kidney in cadmium exposed groups and histopathology of liver, kidney, testis and heart were studied. Also, interaction of lead and cadmium with zinc and copper in liver, kidney, heart, testis and brain was assessed. The present study revealed significant alterations in body weights, haematology, sero-biochemical parameters, antioxidant profile, ATPases, tissue lipid profile, neurotransmitter, CYPd50, glycogen, GGPD, weights of testes, testicular LDH, sperm count, and concentration of zinc and copper in toxic control groups 3, 4 and 5 as compared to control and NAC-treated groups. The toxic combination (Pb + Cd) group 5 showed significant alterations in most of the parameters studied as compared to Pb alone and Cd alone administered groups. All the NAC-treated groups revealed significant improvement in all the parameters. The histological studies of liver, kidney, testis and brain revealed marked changes in toxic control groups, while therapeutic groups revealed mild changes or no pathologically significant changes. Groups 1 and 2 were devoid of any alterations. The electron microscopy of kidney revealed marked alterations in kidney architecture in groups 4 and 5, while groups 7 and 8 revealed better architecture. The results of the investigation revealed that lead, cadmium' and their combination induced toxicity to the biological system due to the excess generation of free radicals and impairment of antioxidant defenses. Toxic effects were more pronounced in the group that received a combination of lead and cadmium suggesting positive toxicodynamic interaction. Use of NAC countered the adverse effects of lead and cadmium induced toxicity to a major extent suggesting its antioxidant potential owing to replenishment of tissue pool of GSH. Further, NAC administration reduced the extent of accumulation of lead and cadmium in various tissues.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.