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Open Access
SUB-ACUTE TOXICITY STUDIES ON UNPROCESSED AND PROCESSED PONGAMIA PINNATA SEED CAKE IN RATS
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2013-02) SIRISHA, K; KALA KUMAR, B.D.P(MAJOR); GOPALA REDDY, A; MADHAVA RAO, T; ANJANEYULU, Y
ABSTRACT : The present study was aimed to evaluate the toxic and protective effects of unprocessed and processed Pongamia pinnata seed cake in rats, which were divided into 3 groups as follows: Group 1: sham control, group 2: unprocessed Pongamia pinnata seed cake included at the level of 9% in the feed (toxic control) and group 3: processed (solvent extracted-isopropyl alcohol) Pongamia pinnata seed cake (detoxified cake) included at the level of 9% in the feed. Average body weights were recorded at weekly intervals and on 28th day, organs were collected for estimation of TBARS, protein carbonyls and GSH in kidney, liver and testes homogenates and estimation of epididymal sperm count from testes collected. Sero-biochemical parameters like ALT, total proteins & globulins, total cholesterol, HDL & LDL cholesterol, creatinine and LDH were estimated at fortnight intervals. Haemotological parameters (RBC, WBC, Hb and PCV) were also estimated at fortnight intervals. Serum troponins, PHA assay and testicular LDH were estimated at the end of the experiment. Histopathology of heart, liver, kidney, spleen and testis was also studied at the end. Mean body weight gain, GSH, total proteins and globulins, PHA assay and sperm count were significantly (P < 0.05) decreased in toxic control group (group 2), while TBARS, protein carbonyls, serum LDH, intra-testicular LDH, serum ALT, creatinine, total cholesterol and serum troponins were significantly (P < 0.05) increased in group 2. There was no significant difference in TEC, TLC, Hb, PCV, HDL cholesterol, LDL cholesterol, mean body weight gain (in the 1st week) in group 2. Group 1 did not reveal any abnormalities on histopathology. Group 2 showed interfibrillar haemorrhages, congestion and edema with disruption of cardiac myofibres in heart, marked degenerative changes in tubular epithelial cells and marked dilatation of tubules in kidney, marked central vein congestion and marked bile duct hyperplasia in liver, congestion and thickening of trabecular arteries in spleen and finally marked congestion and edema with disrupted cell wall in seminiferous tubules of testes. Group 3 showed mild lesions in heart, kidney, liver, spleen and testis. From this study, it is concluded that unprocessed Pongamia pinnata seed cake induces toxicity to heart, kidney, liver, spleen and testes, and group 3 showed restoration in all the parameters studied, suggesting reduced toxic potential of processed seed cake.
Open Access
ENDOCRINE DISRUPTING ACTIONS OF CADMIUM AND EXPERIMENTAL EVALUATION OF PROTECTION BY GREEN TEA EXTRACT
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2014-05) SHIVAKUMAR, PABBATHI; GOPALA REDDY, A(MAJOR); SRINIVASA RAO, G; ANJANEYULU, Y; RAMANA REDDY, Y; UDAYA KUMAR, M
ABSTRACT : An experimental study was conducted to evaluate the neuro-endocrine disrupting actions of cadmium and the effect of cadmium on the progeny that were born to cadmium exposed rats and to evaluate the protective role of green tea on neuro-endocrine disrupting actions of cadmium in Sprague dawley rats. Rats were randomly divided into 4 groups of 30 rats in each (male rats =12, female rats=18).Group 1 served as Sham control Group 2 treated with CdCl2, Group 3 treated with Green tea extract treatment and Group 4 Cd + green tea extract treatment. Blood was collected from all the groups at monthly intervals for analyzing sero-biochemistry (blood glucose, total cholesterol, HDL-cholesterol, triglycerides, total protein and albumin, biomarkers of cardiovascular, hepatic and renal pathology, and hormonal profile (thyroid profile, sex hormones). The key enzymes concerned with metabolism were assayed. Immune status was studied at the end of 3rd month by phytohaemagglutinin assay. Rats were subjected to neuro-behavioural studies at the end (Elevated plus maze and Morris water maze). Epididymal sperm count in males and estrous cycle pattern in females were studied. At the end of 3 months, 12 rats (6 males and 6 females) from each group were sacrificed to collect various organs and endocrine glands and subjected them to biochemical, histological and electron microscopic studies. Cadmium concentration was estimated in all the treated groups in kidney, testes, liver and brain at the end of 3 months. In all the groups, twelve (12) females were mated at the end of three months with male rats belonging to respective groups/treatments and the treatment was continued till 17th day of gestation. 50% of the pregnant rats in the respective groups were sacrificed on day 19 to study skeletal and soft tissue developmental anomalies and the rest were allowed to normal delivery. The pups of F1 generation from all the groups were kept till weaning (post-natal day 21) and were subjected to sero biochemical, neurobehavioural studies andthyroid hormone profile were estimated. There were significant alterations in sero-biochemistry biomarkers of cardiovascular, hepatic and renal pathology and hormonal profile thyroid profile, group 2 as compared to group 1.Treatment group revealed significant improvement in all the parameters as compared to group 2, while the combination treatment group 4 was found better The histological studies in group 2 revealed marked changes in all the organs studied, while groups 4 revealed moderate changes and groups 1 and 3 revealed no pathologically significant changes. The electron microscopy of kidney, testis and thyroid revealed marked alterations in architecture in group 2, while groups 4 revealed better architecture. There were no significant alteration in the TEM samples of the offspring and there were no skeletal abnormalities in the offspring as evidenced by skeletal staining. The results of the study revealed neuro-endocrine disrupting actions of cadmium and protctive role of green tea in cadmium toxicity. Further studies are warranted to know in detail on the endocrine disrupting actions of cadmium and protective role of green tea at various concentrations.
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, A
ABSTRACT: 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.
Open Access
IN VITRO STUDIES ON MODULATION OF CONTRACTILE AND RELAXANT RESPONSES OF SHEEP TRACHEAL SMOOTH MUSCLE BY MORIN
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2016-07) MURALI KRISHNA, M; BHARAVI, K(MAJOR); Ravi Kumar, P; Satheesh, K
ABSTRACT: Morin (3,5,7,2,4 - pentahydroxyflavone), belongs to the flavonoid group of polyphenolic compounds found in wood of old fustic, in osage orange, white mulberry, fig, almond and sweet chestnut, and many other herbs and fruits used as herbal medicines. Morin has been the subject of a number of experimental studies dealing with its pharmacological activities, such as antioxidant properties, inhibition of lipid peroxidation, anti cancer activity, anti-inflammatory activity, cardiovascular protection and also a prooxidant action. Despite the enormous interest in flavonoids as potential therapeutic agents, their biological activity mechanisms are poorly understood and largely unknown, but different types of possible biochemical events are involved. In view of this, the present study is designed to know the modulatory effect of morin hydrate on contractile and relaxant responses of various pharmacological agents like acetylcholine and atropine, histamine and cetirizine, KCl and nifedipine in sheep tracheal smooth muscle in vitro. Mean EC50 values for contractile response of acetylcholine obtained in the absence of morin hydrate were 3.38×10-7M (5×10-9M to 15×10-6M) and in presence of morin hydrate it was increased to 6.02×10-7M (5×10-9M to 15×10-5M) with a significant right shift. Mean EC50 values for contractile response of histamine obtained in the absence of morin hydrate were 1.12×10-6M (5×10-9M to 30×10-5M) and in presence of morin hydrate it was increased to 2.63×10-6M (5×10-9M to 30×10-5M) with a significant right shift. Mean IC50 values for relaxant response of atropine, cetirizine and nifedipine in comparison with morin hydrate in sheep tracheal smooth muscle were 2.23×10-8M (5×10- 9M to 5×10-6M), 3.46×10-8M (5×10-9M to 15×10-6M), 6.30×10-6M (5×10-9M to 30×10- 5M) and 6.30×10-7M (5×10-9M to 15×10-6M), 1.58×10-6M (5×10-9M to 5×10-5M), 2.04×10-5M (5×10-9M to 15×10-4M) respectively. The study revealed that morin hydrate has relaxant effect on the sheep trachealis smooth muscle in vitro.
Open Access
STUDIES ON DIAGNOSTIC AND PROGNOSTIC FACTORS IN BOVINE FORESTOMACH DISORDERS
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI – 517 502. (A.P) INDIA, 2015-09) RAMU, VEGIREDDI; RAGHUNATH, M(Major); HARI KRISHNA, N.V.V; VENKATA NAIDU, G
Abstract : The present study was conducted on 22 clinical cases of cattle and buffaloes affected with forestomach disorders that were referred for treatment to the Department of Veterinary Surgery & Radiology, NTR CVSc, Gannavaram. Detailed signalment, history, clinical observations, haematological and biochemical estimation were recorded and evaluated. The radiographic features of reticulo diaphragmatic hernia, reticulophrenic adhesions and foreign bodies in the reticulodiaphragmatic area were studied. Based on the tentative diagnosis, the animals were divided into two groups as in Group I (N = 6), animals with forestomach disorders that responded to medical treatment were included and those that failed to respond and surgically treated were included in Group II. In turn, Group II was divided into Group II A (N = 6) where forestomach disorders not induced by foreign bodies were included and Group II B (N = 10) where forestomach disorders induced by foreign bodies were included. Majority of the animals were characterized by partial to complete anorexia, rumen atony to hypermotility, recurrent to persistent tympany, hard and scanty or loose dung and most of the animals with pericarditis cases were characterized by brisket edema, bilateral jugular engorgement with abnormal heart sounds. Hypochloremia, hypocalcemia, hypophosphatemia and hyponatremia were mostly evident in animals with all these disorders. Increased peritoneal fluid quantity was observed in cases of pericarditis. Ruminal fluid pH, rumen protozoan motility, radiography and exploratory laparorumenotomy have good diagnostic value in forestomach disorders. Laparorumenotomy has both diagnostic and therapeutic value in cases of rumen impaction, omasal impaction, traumatic reticulitis and diaphragmatic hernia cases. Delayed presentation, hypochloremia and loss of tonocity of reticulo-omasal orifice were poor prognostic signs in omasal impaction cases. High neutrophil count, low haemoglobin levels, severe hypochloremia and extensive ring size were poor prognostic factors in cases of DH. Potential penetrating metallic foreign bodies crossing diaphragm and pointing towards thorax, with high neutrophilia with left shift had poor prognostic value in traumatic pericarditis cases.
Open Access
IN VITRO STUDIES ON MODULATION OF CONTRACTILE AND RELAXANT RESPONSES IN SHEEP TRACHEAL SMOOTH MUSCLE BY CADMIUM
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI – 517 502. (A.P) INDIA, 2014-10) HEMANTH KUMAR, REGULAVALASA; Ravi Kumar, P (Major); Srinivasa Rao, G; Sailaja, N
ABSTRACT: Cadmium is an environmental contaminant of increasing importance. Of various heavy metals, cadmium is known to have significant effect on the bronchial structures due to its portal of entry. In view of this, the present study is designed to know the modulatory effect of cadmium on the contractile or relaxant responses of various pharmacological agents like acetylcholine, salbutamol, sildenafil and sodium nitro- prusside in sheep tracheal smooth muscle in vitro. The control strips and the cadmium chloride treated strips were exposed to the contractile and relaxing agents like acetylcholine iodide, salbutamol, sildenafil and SNP in cumulative doses. At the end of each study the treated and the normal tissues were preserved for histopathological studies. Mean EC50 values for contractile response of acetylcholine obtained in the absence of cadmium chloride was 4.42 x 10-6M (1.9×10-8M to 1.9×10-5M) and in the presence of cadmium chloride it was increased to 6.43 x 10-5M (1.9×10-8M to 6.3×10-3M) with a significant right shift. Mean IC50 values for relaxant responses of salbutamol, sildenafil and sodium nitroprusside obtained in the normal sheep tracheal smooth muscle were 2.63 x 10-4M (1.9×10-8M to 6.3×10-3M), 3.66 x 10-4M (1.9×10-8M to 1.9×10-4M) , 8.64 x 10- 5M (1.9×10-8M to 1.9×10-4M) respectively. Cadmium chloride treatment resulted in a significant decrease in the mean IC50 values of salbutamol, sildenafil and sodium nitroprusside to 5.82 x 10-5M (1.9×10-8M to 1.9×10-4M), 2.17 x 10-5M (1.9×10-8M to 6.3×10-3M) and 4.48 x 10-6M (1.9×10-8M to 1.9×10-4M) respectively. Inhibition of acetylcholine induced contraction and promotion of relaxation mediated by salbutamol, sildenafil and sodium nitroprusside in isolated sheep tracheal smooth muscle by cadmium might be due to the blockade of Ca2+ channels. Mononuclear cell infiltration was seen in cut sections of trachea after cadmium treatment for 30 min. These cells are known to produce NO, which also might have aided the smooth muscle relaxation. The decreased [Ca2+]i and mononuclear cell mediated NO production together might be responsible for the relaxant effect of cadmium on sheep tracheal smooth muscle
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, A
ABSTRACT : 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.
Open Access
PHARMACOKINETICS OF CITRATE MEDIATED NANOSILVER IN MURRAH BUFFALO CALVES
(SRI VENKATESWARA VETERINARY UNIVERSITY , TIRUPATI – 517502. (A.P.) INDIA, 2014-02) SIVA MANOHARA RAO, V.K; Adilaxmamma, K (Major); Alpha Raj, M; Eswara Prasad, P; Prasad, T.N.V.K.V
ABSTRACT : An experiment was conducted to study the pharmacokinetics of citrate mediated nanosilver particles in Murrah buffalo calves. The study was conducted on eight healthy male murrah buffalo calves of 6-12 months age at a local farm in Pulivendula, Y.S.R. Kadapa district of Andhra Pradesh. Eight male murrah buffalo calves were randomly divided into two groups. Group I and II were administered Citrate mediated nanosilver @ 0.438 mg through intra-venous (i.v) and intra-muscular (i.m) route respectively. Whole blood samples were collected at time intervals of 0 min, 10 min, 30 min, 1 h, 2 h, 4 h, 8 h, 12 h, 16 h, 24 h and 36 h. The whole blood samples were subjected to acid digestion and carried out the quantification of AgNPs and other elements like Calcium, Manganese, Sodium and Potassium using ICP-OES. Silver nanoparticles were synthesized from silver nitrate using Sodium citrate. The final concentration of silver nanoparticles was 73 ppm with a size of 77.97 nm and peak absorption at 420 nm. The AgNPs were irregular and spherical in shape with uniform distribution with a zeta potential, conductivity and electrophoretic mobility of - 40 mV, 3.198 mS/cm and -0.000311 cm2/Vs respectively. FT-IR analysis of AgNPs showed transmission peaks at 3437.86, 2113.69, 1637.57and 1189.32 cm-1. The in vitro antibacterial activity of AgNPs against Staphylococcus aureus showed a minimum inhibitory concentration of 3.02 μg/mL. In vitro cytotoxicity on mouse splenocytes showed a viability of more than 93% at 9.38 ppm, which was reduced to 77.63% at 18.75 ppm and to 70.37% at 37.5 ppm concentration. In acute oral toxicity testing in rats, at a test dose of 487 μg/ kg b.wt. no mortality or adverse effects were observed. The whole blood concentrations of AgNPs administered through intravenous route showed rapid distribution, reaching undetectable level by 1 h and remained so until 2 h. After, 4 h there was considerable redistribution into circulation reaching a peak concentration of 0.9225±0.2265 μg/mL at 24 h followed by a steady decrease from 36 h.The whole blood concentrations of AgNPs administered through intramuscular route indicated that AgNPs were not absorbed into the circulation at any time interval during the study. AgNPs exhibited significant interactions with Calcium (p=0.784; p<0.01), Manganese (p=0.897; p<0.01) and Potassium (p=0.756; p<0.01). Further, the correlation with potassium was not considered since corresponding significant changes in potassium concentrations were not found at any point of time during the study. The initial phase of AgNPs concentrations from 10 min to 1 h followed one compartmental model of distribution (Y=0.222e-1.22x; R2 = 0.963), with a rapid elimination rate constant (K10) of 1.2238 h-1 and short T1/2 of 0.5663 h. As the whole blood AgNPs concentrations in total could not be fitted into any compartmental model, non-compartmental analysis (NCA) was used. NCA revealed that Cmax and the corresponding Tmax were 0.9225 μg/mL and 24 h respectively. The area under the curve (AUC0-t) was 17.1604 μg/mL*h and AUCt-∞ was 40.8129 μg/mL*h. The mean residence time (MRT) was found to be 51.7788 h with a terminal elimination rate constant (λ) of 0.0279/h and terminal half life (T1/2) of 24.8404 h. In the intramuscular route, pharmacokinetic analysis could not be performed since no absorption of AgNPs was detected. In conclusion, since Citrate mediated silver Nanoparticles exhibited unpredictable pharmacokinetics, erratic redistribution, very long MRT, slow elimination rates and considerable interactions with major elements in blood like calcium and manganese, it is concluded that citrate mediated nanosilver is not suitable for systemic administration either through intravenous or intramuscular route in buffaloes. However, further studies are required to validate the present finding.

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