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ThesisItem Open Access COMBINED EFFECT OF DIETARY AFLATOXIN AND FUMONISIN ON SERUM BIOCHEMICAL PROFILE IN BROILERS AND THEIR AMELIORATION USING ABSORBANTS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2006-02) JAYASRI, K; NARASA REDDY, G.V(MAJOR); SETTY, O.H.; RAJASEKHAR REDDY, A; REDDANNA, PThesisItem Open Access ISOLATION OF BUFFALO URINARY PROTEINS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2007-09) RAGA SUDHA, MEDICHERLA; Ashok, VASILI(MAJOR); Viroji Rao, S.T.; Narasimha Reddy, YABSTRACT: Presence of proteins in urine is considered pathological, but still certain proteins are excreted in minute quantities into the urine of healthy animals. The normal proteins in the urine are uroplakins (15 to 47kDa) and uromodulin (85 to 90kDa) which are produced by the urinary bladder and the thick ascending limb of Henle’s loop (kidney) respectively. The tissue specific expression of these proteins in the urinary system formed the basis for transforming the urinary system into a bioreactor to produce proteins of economic importance in the urine. In the present day world, the idea of transforming urinary system as a bioreactor and urine as an expression system has growing importance. So, the objective of the present study is isolation and quantification of these normal buffalo urinary proteins. The outline of the work is collection of urine from apparently healthy lactating Murrah buffaloes. Precipitation of proteins from the urine is carried out by protein precipitating agents like acetone, ammonium sulphate and sodium chloride. These precipitated proteins were analyzed by SDS-PAGE. Coomassie blue stained gel revealed a prominent and consistent protein band of molecular weight 85kDa in all the precipitated samples which could be uromodulin, the protein of our interest. Silver nitrate stained gels revealed low molecular weight protein bands (15, 27 and 47kDa) in all precipitated samples in addition to the high molecular weight band, which could be uroplakins. The consistent production of uromodulin by the animal into the urine gives an idea of transforming the kidney into a bioreactor. The reason for inconsistent occurrence of various low molecular weight proteins could be due to the dilution effect of liquid intake by the animal, diet, temperature, climate and various physiological factors which are yet to be ruled out. Quantification of protein precipitates of urine samples and urine as such of 10 different healthy buffaloes was done. The mean protein yields (mg) of urine and precipitates of acetone, ammonium sulphate and sodium chloride are 0.96±10.20, 10.60±0.39, 1.50±0.04 and 1.91±0.13 respectively. Acetone precipitation yielded significantly (p<0.001) higher quantity of protein precipitate. However, the banding pattern is not commensurate with the amount of precipitate. This could be the result of interference of salts and metabolic wastes with the Lowry’s method, yielding higher values. Ammonium sulphate precipitated more number of proteins, evidenced by the gel replicates, but at higher concentration. Sodium chloride selectively precipitates almost a single protein and is required in small quantities when compared to ammonium sulphate for producing good banding pattern. It is concluded that acetone precipitates higher amounts of protein quantitatively. Ammonium sulphate precipitates more number of proteins, where as sodium chloride precipitation yields almost a single protein. However, sodium chloride method has the advantage of selective precipitation over acetone and ammonium sulphateThesisItem Open Access PROTEIN PROFILE OF POULTRY KIDNEY(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2007-09) KALYANI, P; ASHOK, VASILI(MAJOR); RAJASEKHAR REDDY, A; NAGAMALLESHWARI, YABSTRACT : The extensive prevalence and escalating incidence of Chronic Renal Failure and End Stage Renal Disease worldwide necessitates renal tissue engineering as a practical solution to meet the organ demand for renal transplantation therapy. Renal tissue engineering is a multidisciplinary field, which is supported on three pillars- cells, scaffolds and signaling biomolecules. A critical understanding of the role of signaling biomolecules is necessary for tissue engineering because these biomolecules are differentially expressed with specific spatio-temporal distribution. Signalling biomolecules and signal transduction pathways are highly conserved between species during millions of years of evolution, which engenders the scope for probing into mammalian kidney biology by using lower organisms as models. Gallus gallus (chicken) is a suitable animal model to study the role of proteins in renal organogenesis, owing to the availability of extensive literature on embryogenesis and the ease of embryo retrieval. Metanephric kidneys were collected from different developmental stages of Vanaraja breed chick embryos viz., on 7th , 9th , 11th , 13th , 15th , 17th , 19th days , day old chicks and adult cock in ice cold PBS. Embryonic kidneys and kidneys of day old chicks were dissected under diascopic stereo zoom microscope. The kidney tissues were homogenized, centrifuged and supernatants were analyzed by SDS PAGE. Prominent, differential protein expression was evident upon comparing the protein bands of tissue extracts. In total, 22 prominent differentially expressed proteins were identified on careful examination of banding pattern from the tissue extracts of different developmental stages. A single high molecular weight protein band was seen only in the adult sample. Some of these prominent protein bands were identified to be MMP9 (97 kDa), nephronectin (79 kDa), osteopontin (76 kDa), MMP2 (72 kDa), pax 8 (57 kDa), pax 2 (50 kDa), BMP 4 (45 kDa), actin (43 kDa), FGF2 (23kDa), pleitrophin (21kDa), activin 1 (20kDa), endostatin (15kDa) based on their molecular weights and temporal distribution. Further confirmation of the identity of these proteins could be done by in vitro cultural tests, protein sequencing and other functional tests. The present study reveals a prominent, differential protein expression at various developmental stages in kidney organogenesis. Based on the banding pattern, it is proposed that most of the architectural design for kidney organogenesis is framed by the end of embryonic day 15 and after that it is refined to perfection to carry out various biological activities. Research on the histology and immuno-histochemistry of the kidney tissue during various developmental stages can confirm the present findings.ThesisItem Open Access EFFICACY OF ACTIVATED CHARCOAL AND YEAST CULTURE ON AMELIORATION OF COMBINED TOXICITY OF AFLATOXIN AND T-2 TOXIN IN BROILERS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2007-01) RAGHAVENDRA SRIKANTH, N; NARASA REDDY, G.V(MAJOR); SETTY, O.H.; RAJASHEKAR REDDY, AABSTRACT : Aflatoxin and T-2 toxin were produced from Aspergillus parasiticus and Fusarium sporotrichioides (F. tricinctum) respectively and quantified using TLC. The toxins were mixed with broiler feed to attain required concentrations. Four diets for broilers were prepared diet 1 – basal diet (control), diet 2-basal diet + aflatoxin (1 ppm) + T-2 toxin (1 ppm), diet 3 – basal diet + aflatoxin (1 ppm) + T-2 toxin (1 ppm) + activated charcoal (0.4%), diet 4 – basal diet + aflatoxin (1 ppm) + T-2 toxin (1 ppm) + activated charcoal (0.4%) + yeast culture (0.1%). These four diets were fed to 4 groups of day old broiler chicks with four replicates of eight birds in each replicate, in a completely randomized design for six weeks. The body weight gains were significantly (P<0.01) lower on diet 2 containing aflatoxin and T-2 toxin. The body weight gains increased gradually on diet 3 containing activated charcoal and on diet 4 containing activated charcoal and yeast culture. Feed consumption was significantly (P<0.01) higher on basal diet compared to all other diets, which were at par. Feed conversion ratio was the highest on diet 2 compared to other diets which were at par. On diet 2 containing aflatoxin and T-2 toxin, the biochemical profile revealed elevated serum enzyme activities of AST, ALT, ALP, GGT indicating liver toxicity. These studies also recorded decreased levels of serum proteins, glucose, cholesterol, triglycerides, calcium, uric acid and increased levels of serum bilirubin. Increased serum creatinine and BUN recorded indicates toxicity to kidney. The studies on liver and kidney function tests indicate possible damage of these organs in the chicks fed on diet 2. Degenerative changes in liver, kidney and lymphoid depletion in bursa of Fabricius, disruption of cardiac muscle fibres, and disruption of intestinal villi were observed on histopathology in birds fed with aflatoxin and T-2 toxin supporting the biochemical study. The birds on diet 3 showed lower body weight gains and feed consumption compared to that of control diet. Efficiency of feed utilization on diet 3 was comparable to that of diet 1. The biochemical parameters like AST, ALP, GGT, serum calcium, serum uric acid were moderately improved but there was no improvement in creatinine and bilirubin compared to diet 2. There was mild improvement in serum proteins cholesterol, glucose and triglyceride levels compared to diet 2. Complete alleviation of toxic symptoms was noticed in terms of serum ALT and BUN. Broilers on diet 4 showed an increase in body weight gains compared to diet 2. Feed conversion ratio on diet 4 was comparable to that on diet 1. The biochemical profile of cholesterol total protein, glucose, uric acid, calcium and triglyceride showed moderate improvement while the other parameters like ALP, bilirubin, ALT, GGT and AST were comparable to those of control diet. Histopathological studies were comparable to those of control group fed on toxin free diet. These results indicate that aflatoxin and T-2 toxin in the diet caused deteriorated effects on growth, biochemical profile and caused damage to vital organs. The combination of activated charcoal and yeast culture was more effective in counteracting the combined toxicity of aflatoxin and T-2 toxin compared to the activated charcoal alone.ThesisItem Open Access DETOXIFICATION OF CITRININ AND FUMONISIN IN FEED AND ITS EFFECT ON SERUM BIOCHEMICAL PROFILE IN BROILERS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2007-01) ASHOK KUMAR, DEVARASETTI; ASHOK, VASILI(MAJOR); NARASA REDDY, G.V; ANJANEYULU, YABSTRACT : Citrinin and fumonisin toxins were produced from Pencillium citrinum and Fusarium moniliforme respectively and both were quantified using Thin Layer Chromatography (TLC) method. The toxins citrinin @ 50 ppm and fumonisin @ 10 ppm were mixed in broiler feed to attain required concentrations. Four diets for broilers were prepared diet 1 – basal diet (control), diet 2 – basal diet + citrinin (50 ppm) + fumonisin (10 ppm), diet 3 – basal diet + citrinin (50 ppm) + fumonisin (10 ppm) + activated charcoal (0.4%) and diet 4 – basal diet + citrinin (50 ppm) + fumonisin (10 ppm) + activated charcoal (0.4%) + lyophilized yeast culture (0.1%). Each diet was fed to a group of day old broiler chicks with four replicates having eight birds in each replicate in a completely randomized design for six weeks. The weekly body weights and weekly body weight gains were significantly (P < 0.01) lower on diet 2 containing citrinin (50 ppm) + fumonisin (10 ppm). The weekly body weights and weekly body weight gains increased gradually on diet 3 containing activated charcoal. Further significant (P < 0.01) improvement was recorded on diet 4 containing activated charcoal and yeast culture. Feed consumption was significantly higher (P < 0.01) on basal diet compared to other test diets. Feed conversion ratio was the highest on diet 2 when compared to other test diets. On diet 2 containing citrinin and fumonisin, the biochemical profile revealed elevated serum enzyme activities of AST, ALT, ALP, GGT indicating liver toxicity. These studies also recorded decreased levels of serum proteins, albumin, glucose, cholesterol, triglycerides, calcium, uric acid and increased levels of serum bilirubin. Increased serum creatinine and BUN recorded indicates toxicity to kidney. These studies on liver and kidney function tests indicate possible damage of these organs in the chicks fed on diet 2. Further, supporting the biochemical study, degenerative changes and central venous congestion in liver and marked degenerative changes in kidney and lymphoid depletion in bursa of Fabricius, disruption of cardiac muscle fibres, disruption of intestinal villi were observed on histopathology in birds fed with citrinin and fumonisin. The birds on diet 3 containing citrinin, fumonisin and activated charcoal showed significant (P < 0.01) improvement in body weight gains and feed consumption. Efficiency of feed utilization on diet 3 was comparable to that of diet 1. The serum enzymes like AST, ALT, ALP and GGT were markedly improved and serum creatinine, BUN, calcium, serum uric acid were moderately improved when compared to diet 2. There was mild improvement in serum proteins, cholesterol, glucose and triglyceride levels compared to diet 2. All other biochemical parameters and histopathological findings showed improvement compared to toxin fed group (diet 2). This indicates that activated charcoal (0.4 %) had partial amelioration on toxic effects of citrinin and fumonisin. The birds fed on diet 4 recorded the serum enzymes like AST, ALT, ALP and GGT in the liver nearer to the control group (diet 1). All the biochemical parameters and histopathological findings were also nearer to that of the control group and within the normal range indicating that activated charcoal and lyophilized yeast culture had a complementary effect in ameliorating the toxic effect of citrinin and fumonisin. These results indicate that activated charcoal (0.4 %) could adsorb and ameliorate the toxins (citrinin and fumonisin) to some extent and activated charcoal (0.4 %) and lyophilized yeast culture (0.1 %) showed complementary effect in amelioration of the toxic effect caused by citrinin and fumonisinThesisItem Open Access COMBINED EFFECT OF DIETARY AFLATOXIN AND CITRININ ON SERUM BIOCHEMICAL PROFILE IN BROILERS AND THEIR AMELIORATION USING ADSORBENTS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2005-12) PRIYADARSHINI, C.H.; NARASA REDDY, G.V(MAJOR); SETTY, O.H.; ANJANEYULU, YABSTRACT: Aflatoxin was produced from Asprgillm parasiticus and cihinin was produced from Penicillium cirri- ,dtd h were quantifd using thin layer chromatography. The toxins, aflatoxin at the rate of lppm and citrinin at the rate of 25 ppm, were mixed in broiler feed. Activated charcoal (0.4%) and/or lyophilized yeast culture (0.2%) were added as adso&ents. Four diets for broilers were prepared - Diet 1- basal diet (control). Diet 2- basal diet added with aflatoxln (I ppm) and citrinin (25ppm). Diet 3- basal diet added with aflatoxin (Ippm), citrinin (25 ppm) and activated charcoal (0.4%) and Diet 4- basal diet added with aflatoxin (Ippm), citrinin (25ppm). activated charcoal (0.4%) and lyophilized yeast culture (0.2%). These four diets were fed for 6 weeks to four group of broiler chicks with four replications of eight birds in each gnwrp using completely randomized design. The body weight gains. feed consumption and feed conversion ratio were significantly (Pa.01) lower on diet 2 containing aflato.. in and citrinin. These values were significantly improved on diet 3 containing aflatoxin. citrinin and activated charcoal. Further significant improvement was nxoded on diet 4 containing aflatoxin. citrinin. activated charcoal and lyophiliztd yeast culrure but these values were significantly lower than control gmup- On diet 2 containing aflatoxin and citrinin. the biochemical profile of liver function and kidney function of birds showed hepatotoxicity and nephrotoxicity respectively as evidenced by increased serum enzyme activities of AST. ALT. alkaline phosphatase and GGT concentration in the liver and increased levels of serum creatinine, blood urea nitrogen and serum uric acid. Other biochemical parameters revealed decreased serum protein, albumin, glmse, cholesterol, triglycerides, calcium and phosphorous but increase in serum bilirubin. By these studies on liver and kidney function tests, chicks fad on diet 2 indicate the possible damage of tissues. Further, supporting the biochemical study, degenerative changes and central vein congestion in liver and marked degenerative changes in kidney, lymphoid depletion in bursa of Fabricius and depleted germinal centres in spleen were observed on histopathologv. On diet 3 containing aflatoxin, citrinin and activated charcoal, the serum enzyme activities of AST, ALT. AP and GGT in the liver were slightly decreased and serum --.. creatinine, blood urea nitrogen and serum uric acid levels were slightly decreased compared to toxin group (diet 2). All other biochemical parameters and histopathological findings showed improvement compared to toxin fed group. This indicates that activated charcoal (0.4%) had partial amelioration on the toxic effect of aflatoxin and citrinin. The birds fed on diet 4 recorded the serum enzyme activities of AST, ALT, AP and GGT in the liver nearer to the control group (basal diet). All the biochemical parameters and histopathological f i n d i i were also nearer to that of the control group and within the normal range indicating that activated charcoal and lyophilized yeast had a complementary effect in ameliorating the toxic effect of atlatoxin and citrinin. These results indicate that activated charcoal (0.4%) could adsorb and ameliorate the toxins (aflatoxin and citrinin) to some extent and activated charcoal (0.4%) and lyophilized yeast cultw <0.2O/') showed complementary effect in amelioration of the toxic effect caused by aflatoxin and citrinin.ThesisItem Open Access CONJUGATION OF ANTIBIOTIC RESIDUES PREVALENT IN MILK WITH CARRIER PROTEIN AND RAISING OF POLYCLONAL ANTIBODIES AGAINST THEM(SRI VENKATESWARA VETERINARY UNIVERSITY , TIRUPATI – 517502. (A.P.) INDIA, 2012-08) SAMPATH KUMAR, B; ASHOK, VASILI (Major); ESWAR PRASAD, P; Krishnaiah, NABSTRACT : Antibiotics are the drugs used to kill or inhibit the growth of bacteria. Presence of antibiotic residues in milk has significant public health importance as they have adverse side effects on the health. The present study was undertaken to produce polyclonal antibodies against the most commonly used antibiotics in large animal practice so that a diagnostic test can be developed to screen milk samples for the presence of antibiotic residues. Ampicillin, ceftiofur, enrofloxacin and gentamycin were conjugated with carrier protein BSA by carbodiimide method in which EDC was used as a crosslinking agent. Successful conjugation was confirmed by SDS PAGE in which antibiotic conjugates revealed high molecular weight protein bands of 90kDa, 85kDa, 78kDa, 74kDa for gentamycin, ampicillin, ceftiofur and enrofloxacin respectively in contrast to 68kDa band of BSA. Sprague Dawley rats aged 7-8 weeks were immunized with the antibiotic-BSA conjugates by subcutaneous route according to the standard immunization schedule. A total of four blood samplings were done from each group (3 animals) as follows: first three samples serially at 15 days time interval after 1st immunization plus 1st booster, 2nd booster, 3rd booster and the 4th sampling one and half month after the third booster. The mean total protein content in the antisera of ampicillin, ceftiofur, enrofloxacin and gentamycin groups were 12±1.15 g/dL, 30±1.20 g/dL, 24±1.73 g/dL and 8.25±0.20 g/dL respectively and in the control group serum it was 6.66±0.01 g/dL. The mean albumin concentrations were 3.44±0.020 g/dL, 3.36±0.173 g/dL, 3.21±0.003 g/dL & 2.93±0.014 g/dL for ceftiofur, ampicillin, enrofloxacin and gentamycin groups respectively compared to 3.36±0.173 g/dL in the control group. The mean globulin content in the antisera of ampicillin, ceftiofur, enrofloxacin and gentamycin groups were 8.64±0.10 g/dL, 26.56±1.18 g/dL, 20.79±0.08 g/dL and 5.32±0.05 g/dL respectively and it was 3.30±0.04 g/dL in the control group. A/G ratios were less than the normal value (>1) in antisera of ampicillin, ceftiofur, enrofloxacin and gentamycin groups. Indirect ELISA was standardized with antibiotic-casein conjugate as coating antigen to detect antibody titres. The optimum antigen concentration was chosen as 20ng/ml (which was in the range of Maximum residue limits) and optimum antiserum dilution was chosen as 1/100 by checker board titration. The antibody titres in the test samples of 4 immunization cycles for each antibiotic were determined by indirect ELISA at various serum dilutions ranging from 1/100 to 1/6400. Cut off value of 0.3 was calculated from PNT base line constructed with M+3SD values of control serum samples. All the four antibiotic-BSA conjugates elicited positive immune response in rats. The immune response in BSA-ampicillin conjugate immunized rats was maximum with the highest antibody titre of 2.577 (OD450) followed by BSA-ceftiofur conjugate immunized rats, BSA-enrofloxacin conjugate immunized rats and BSA-gentamycin immunized rats with OD450 values of 2.072, 1.723, and 0.928 respectively at the antiserum dilution of 1/00. These polyclonal antibodies are the efficient tools for developing immunoassays to detect specific antibiotic residues in biological fluids like milk and urineThesisItem Open Access EFFICACY OF ALOIN MEDIATED PHYTOGENIC NANOSILVER PARTICLES IN EXPERIMENTAL MURINE MASTITIS(SRI VENKATESWARA VETERINARY UNIVERSITY , TIRUPATI – 517502. (A.P.) INDIA, 2013-02) VENKATA CHAITANYA KUMAR, T; ESWARA PRASAD, P (Major); PADMAJA, K; SREENIVASULU, DABSTRACT : An experimental study was conducted to evaluate the efficacy of aloin mediated phytogenic nanosilver particles in experimental murine mastitis. The nanoparticles were synthesized and the synthesized nanoparticles were characterized using UV-visible spectrophotometer, Scanning electron microscope (SEM), DLS (Dynamic light scaterring) Technique and FT-IR analysis and were also evaluated for antimicrobial activity and cytotoxic effects. A total of 40 female mice between 10-15 days of lactation were utilized for the study. The animals were divided into five groups of eight animals each. Group I served as lactating control, groups II to V were inoculated with 20μl of 24h broth culture of S.aureus containing 4.0 x 104 cfu/quarter under ketamine anaesthesia using 33g blunt hamilton needle. After 6h post inoculation, groups III and IV received 20μl of Aloin nano silver (ANS) through intramammary and intra peritoneal routes respectively. Group V received antibiotic Cefepime at a dose rate of 1mg/kg body weight through intra peritoneal route. After 18 h post treatment, serum C reactive protein (CRP) levels, weights of mammary glands, mammary bacterial loads, levels of TBARS, GSH, SOD and CAT, histopathology and ultrastructure studies were conducted. UV Visible absorption spectrum showed a maximum absorption around 200 nm for ANS. SEM images showed that ANS with cubical, rectangular, triangular and spherical morphology of size 287.5 to 293.2 nm. The results of FT-IR analysis indicated the involvement of hydroxyl, carboxyl, amine and nitrile groups in the synthesis of aloin mediated silver nanoparticles. The size of the particles as measured by DLS technique was 67.8 nm. The compound (ANS) showed an MIC of 21.8 ng/ml against Staphylococcus aureus and showed an in vitro spleenocyte viability of more than 80% at the highest concentration of 87.5 ppm per well. The compound was also found to be safe at the limit dose in acute oral toxicity conducted in rats. The bacterial load was significantly reduced in ANS (intramammary), ANS (intra peritoneal) and antibiotic group compared to mastitis control. The levels of C-Reactive Protein were significantly reduced in Aloin treatment groups III and IV compared with antibiotic group V. TBARS was not significantly reduced to the level of normal lactating group, however SOD and CAT levels were lowered by both routes of aloin nanosilver (ANS) treatments. In histopathological sections, significant reduction (98%) in PMN infiltration was observed with ANS than antibiotic (50%). Ultrastructure of mammary gland from aloin treatment groups III and IV indicated moderate progress in the architecture of the secretory epithelial cells showing reconstituting mitochondria and normal inter alveolar septa.ThesisItem Open Access COMPARATIVE EVALUATION OF WOUND HEALING PROPERTIES OF NANOSILVER AND BOSWELLIA OVALIFOLIOLATA MEDIATED NANOSILVER IN WISTAR RATS(Sri Venkateswara Veterinary University, TIRUPATI – 517 502,A.P, 2012-10) APARNA REDDY, N; ESWARA PRASAD, P (Major); PADMAJA, K; ADILAXMAMMA, KABSTRACT : An experimental study was conducted to evaluate and compare the wound healing properties of citrate coated silver nanoparticles and B.ovalifoliolata mediated silver nanoparticles in wistar rats in three wound models. The nanoparticles were synthesized and the synthesized nanoparticles were characterized using UV-visible spectrophotometer, Scanning electron microscope (SEM), DLS (Dynamic light scaterring) Technique and FT-IR analysis and were also evaluated for antimicrobial activity and cytotoxic effects. A total of 90 rats (both sexes) were divided into five groups, each group containing 18 rats and the wound healing activity was evaluated in three different wound models viz., excision, incision and dead space model. Group 1 was treated with normal saline, group 2 with Povidone iodine (1% solution), group 3 with Citrate coated silver nanoparticles (AgNPs) (170 μg per dressing), group 4 received B.ovalifoliolata bark ethanolic extract (50 mg per dressing) and group 5 received B.ovalifoliolata mediated silver nanoparticles (BENS) (85 μg per dressing). The respective concentrations of the drug were prepared as dressings and applied topically. In excision wound model, percentage wound contraction and the periods of epithelialization were recorded. Blood was collected from orbital sinus in EDTA coated tubes and plasma was separated for estimation of Vitamin C, TBARS and protein. Granulation tissue from excision model was processed for histopathological examination. In incision wound model, the wound breaking strength was measured on 10th day. In dead space wound model, the granulation tissue collected on Day 10 was used for the estimation of hydroxyproline, hexosamine, total protein, TBARS, superoxide dismutase, catalase, reduced glutathione and ascorbic acid. UV Visible absorption spectrum showed a maximum absorption between 350-450 nm and around 450 nm for AgNPs and BENS respectively. SEM images showed that AgNPs showed an agglomerated morphology (size 374-718 nm) while, B.ovalifoliolata mediated silver nanoparticles were spherical in shape (size 192.8-346.1 nm). The results of FT-IR analysis indicated the involvement of hydroxyl, carboxyl and amine groups in the synthesis of B.ovalifoliolata mediated silver nanoparticles. The size of the particles as measured by DLS technique was 78.8 and 273.5 nm respectively. The in vitro studies showed that BENS possess highest antibacterial activity against Staphylococcus aureus as evidenced by highest zone of inhibition (17.67 mm) and lowest MIC value (1.77μg/ml). BENS was practically nontoxic at 85 ppm while AgNPs were toxic from 10 ppm onwards. In excision wound model, BENS showed significantly (p<0.05) higher percentage of wound contraction and shortest period of epithelialization compared to control group indicating better wound healing property. The TBARS level was significantly (p<0.05) lower and the levels of vitamin C and protein were elevated in both BE and BENS groups. Histopathological examination of BENS group showed mild cellular infiltration with angiogenesis and more fibroblast proliferation when compared to control. In the incision wound model, BE and BENS groups showed significantly (p<0.05) higher wound breaking strength compared to other groups. In dead space wound model, there was a significant increase in hydroxyproline and hexosamine content with increased catalase and SOD activity and decreased lipid peroxide levels in the granulation tissue of BENS group compared to other treatment groups. Thus in conclusion B.ovalifoliolata mediated silver nanoparticles showed better wound healing properties compared to citrate coated silver nanoparticles and ethanolic extract of B.ovalifoliolata.