Loading...
Thesis
Browse
Search Results
ThesisItem Open Access INTERPLAY OF VITAMIN B6, L-TRYPTOPHAN AND NARINGENIN IN DOXORUBICIN INDUCED CORONARY ARTERY DAMAGE(SRI VENKATESWARA VETERINARY UNIVERSITY, TIRUPATI - 517 502. (A.P.) INDIA, 2024-02) SAI PRATHYUSHA GAJULA; SRIVIDYA .G (MAJOR); SRINIVASA RAO .G; NAVEEN SWAROOP .MThe doxorubicin induced damage in coronary artery was studied initially in porcine coronary artery as in vitro model. The porcine heart was collected immediately after slaughter in chilled Krebs-Henseleit solution (pH-7.4). The Right coronary artery isolated was used for the in vitro experimentation. The RCA was placed in sterile petri dishes in Krebs-Henseleit solution and the total of four treatments with six replications was done. Group I served as control (Normal). Group II served as Doxorubicin control where the RCA was incubated in 40 µM doxorubicin for 3 h. Group III, IV and Group V, RCA were pre-incubated in 30 µM vitamin B6, tryptophan and naringenin for 1h respectively and received 40 µM doxorubicin for 3 h. After completion of treatment with doxorubicin, the tissue was utilized for biochemical and histomorphological examinations. Biochemical tests viz., GSH, SOD, TP and MDA were used to determine the status of oxidative damage. LDH and NO were estimated to determine the extent of cytotoxicity. There is a fall in total protein levels and antioxidant enzymes levels (GSH & SOD) in doxorubicin pre-treated group. Loss of endothelium and inflammatory changes were evidenced in doxorubicin treated coronary artery. There is raise in MDA, LDH and nitric oxide levels in doxorubicin pre-treated group. Pre-treatment with vitamin B6, tryptophan and naringenin caused a protective effect on doxorubicin treated coronary artery and restored the enzymes levels back to normal. Regeneration of endothelium was observed in all three treatment groups upon histomorphological examination. In the in vivo experimentation with rabbits, 36 rabbits of 1.5-2 kg body weight were randomly assigned to 6 groups of six rabbits in each group. Group I served as healthy normal animals and Group II rabbits were given doxorubicin @10mg/kg B,Wt intraperitoneally on 15th and 16th day of study. Group III animals received vitamin B6 @50 mg/kg B.Wt per orally for 14 days. Group IV and V received tryptophan (100 mg/kg B.Wt) and naringenin (100 mg/kg B.Wt) per orally for 14 days. Group VI served as an interplay group where the rabbits received vitamin B6 (50mg/kg B.Wt), tryptophan (100 mg/kg B.Wt) and naringenin (100 mg/kg B.Wt) for 14 days. On day 15 and 16, all groups of rabbits received doxorubicin @10mg/kg B.Wt intraperitoneally. On day 17 the rabbits were sacrificed using xylazine and ketamine anaesthesia and heart samples were collected in chilled Krebs-Hanseleit solution. Half portion of the heart tissues was used to prepare tissue homogenate to study antioxidant enzymes levels and the remaining half to study histomorphological alterations. Blood was collected during sacrifice to study heamatological parameters and the seperated serum was used to estimate calcium, phosphorous, nitric oxide and lactate dehydrogenase. There is a fall in levels of antioxidant enzymes (GSH and SOD), phosphorous, RBC, WBC, haemoglobin, platelets count where as there is elevation of MDA levels, calcium, nitric oxide and LDH levels in the doxorubicin treated rabbits. Gross morphological changes like pale heart, congestion of blood vessels and fatty change and histomorphological alterations like loss of endothelium, infiltration of inflammatory cells, vacuolation of cardiomyocytes were observed in doxorubicin treated rabbits. Pre-treatment with vitamin B6, tryptophan, naringenin and their combination restored the antioxidant enzymes levels, haematological and cytotoxicity markers. Normal architecture of the artery was preserved in these rabbits and gross changes were mild to moderate congestion. Rabbits of Group VI showed less damage which may be due to synergistic effect of vitamin B6, tryptophan and naringenin over doxorubicin damaged heart and coronary artery. Based on the above results, it is concluded that combination of vitamin B6, tryptophan and naringenin offered better protection on doxorubicin induced coronary artery damage.ThesisItem Open Access INTERPLAY OF VITAMIN B1 AND SELECTED ESSENTIAL AMINO ACIDS ON THE ANTIBACTERIAL ACTIVITY AND PHARMACOKINETICS OF CIPROFLOXACIN IN RABBITS(SRI VENKATESWARA VETERINARY UNIVERSITY, TIRUPATI - 517 502. (A.P.) INDIA, 2024-01) ANJANEYULU POTLURI; VENKATA RAO .K.V (MAJOR); SRIVIDYA .G; ASWANI KUMAR .KDrug interaction play a significant role in modulating the pharmacological effect elicited by the drugs in various clinical conditions. Ciprofloxacin, a commonly used quinolone group of antibacterial agent used widely in livestock species as well as in human beings due to its broad spectrum of antibacterial activity. Some of the research findings revealed that vitamin D, Carica papaya alter the kinetic behaviour of ciprofloxacin. This created an interest to study the kinetic behaviour of ciprofloxacin in the presence of vitamin B1 (thiamine) and selected essential amino acid phenylalanine. It is very common that B-complex group of vitamins and dietary supplements with antioxidant activity are prescribed along with antibacterial agents. Thiamine (vitamin B1) is one of B-complex group of vitamin with antioxidant activity, plays a crucial role in energy metabolism, growth, development and functioning of cells. Phenylalanine acts as precursor for tyrosine, dopa (dihydroxyphenylalanine) and catecholamine. The conversion of phenylalanine to tyrosine is facilitated by the enzyme phenylalanine hydroxylase. The sources of phenylalanine include foods like wheat germ, oats, dairy products, and various meats. By keeping this in view the present study was designed to determine the “Interplay of vitamin B1 and selected essential amino acids on the antibacterial activity and pharmacokinetics of ciprofloxacin in rabbits” by microbiological assay. Minimum inhibitory concentration (MIC) of ciprofloxacin using MTCC 443 was calculated by microbroth dilution technique. The MIC end point in the current study was 0.06 µg.mL 1against E.coli MTCC 443. The pharmacokinetic study was conducted in rabbits following single oral administration of ciprofloxacin. Rabbits weighing 2-4 kg randomly divided into 5 groups of 6 each. Group I served as control without any treatment. Group II served as ciprofloxacin control whereas Group III, IV and V rabbits were coadministered with thiamine (80 mg.kg-1), phenylalanine (48 mg.kg-1), combination of thiamine & phenylalanine along with ciprofloxacin (40 mg.kg-1) orally. Blood samples were collected at predetermined time intervals from 0, 0.25, 0.5, 0.75, 1, 2, 4, 6, 8, 12 h and plasma was used for estimation of ciprofloxacin through bioassay. Results obtained from the experiment revealed that rabbits of Group III, IV & V showed improved levels of Cmax are 4.74±0.17, 3.00±0.22, 2.18±0.12 µg.mL-1 and AUC are 20.30±5.19, 10.09±1.48, 12.85±1.34 µg.h.mL-1 respectively when compared to values of Cmax and AUC are 2.10±0.14 and 5.90±0.81 µg.h.mL-1 in Group II rabbits respectively. Among the different interventions thiamine coadministered with ciprofloxacin showed improved antibacterial activity, bioavailability and duration of action of ciprofloxacin in rabbits. There is no much significant role of combination of thiamine and phenylalanine (Group V) coadministration with ciprofloxacin when compared to only thiamine coadministered with ciprofloxacin (Group III) and only phenylalanine with ciprofloxacin (Group IV) on kinetic behaviour of ciprofloxacin in rabbits. PK-PD integration of the present study revealed that Cmax:MIC>8, AUC:MIC>125 in Group III, IV and V which implies that the coadministration of vitamin B1 and phenylalanine and their combination improves the antibacterial activity and reduce the development of resistance at the selected dose of ciprofloxacin.ThesisItem Open Access STUDIES ON THE HEPATOPROTECTIVE EFFECT OF STOLEPHORUS COMMERSONNII FISH EXTRACT AND FLAVONOID RUTIN IN RATS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2023-03) NAGA MOUNIKA PERAVALI; RAVI KUMAR .P (MAJOR); BHARAVI .K; Sudhakar .KLiver plays a major role in the metabolism, detoxification and excretion of xenobiotics. As a consequence, hepatocytes are exposed to high concentrations of these toxic substances, which can lead hepatoxicity, liver dysfunction, and even total organ failure. The main pathological pathway involved in majority of the liver disorders is oxidative stress and its associated lipid peroxidation. Inspite of tremendous advancements in modern medicine, natural antioxidants and hepatoprotective agents are widely considered to be superior over conventional drugs due to the adverse effects associated with the latter. Experimentally, carbon tetrachloride (CCl4) is commonly used to induce oxidative stress related liver injury in animals. Hepatotoxicity induced by CCl4 involves the formation of trichlormethyl free radical (CCl3•) by cytochrome P-450 enzyme system, that is further transformed to highly reactive trichloromethylperoxy radical (CCl3OO•) in the presence of oxygen. This highly reactive radical then initiates a chain reaction of lipid peroxidation by attacking polyunsaturated fatty acids, disrupts calcium homeostasis and eventually kills cells. Stolephorus commersonnii is a marine water fish enriched with high amounts of essential amino acids (lysine, isoleucine, threonine, and serine), omega-3 polyunsaturated fatty acids (EPA, DHA), and minerals (iron, copper, zinc, sodium, and potassium). It has been proven to possess several health benefits due to its high nutritional and therapeutic significance. Rutin is a flavonol glycoside that is widely distributed in nature in various vegetables and fruits such as the passion flower, buckwheat, green asparagus, apple, oranges, onions and tea. It is known for its antioxidant and inhibitory effects on generation of reactive oxygen species and membrane lipid peroxidation. The current work was planned to study the hepatoprotective effect of chloroform: methanolic extract of Stolephorus commersonnii fish, flavonoid rutin, and their co-administration against CCl4-induced toxicity in adult male Wistar albino rats. The animals were randomly assigned to five equal groups, with six animals in each. Group I (control) animals were treated with 1% DMSO (vehicle) orally for 3 weeks and olive oil @ 1ml/Kg b.wt. IP twice a week in 2 nd and 3rd week. Animals in Group II which served as toxic model received CCl4 @ 1ml/Kg b.wt in olive oil (1:1) IP twice a week in 2 nd and 3rd week. Group III animals were administered with Stolephorus commersonnii fish extract (SCFE) orally @ 300 mg/Kg b.wt. in 1% DMSO for three weeks, along with CCl4 as in Group II. Group IV animals were treated with rutin @ 20mg/Kg b.wt. in 1% DMSO orally for 3 weeks along with CCl4 as in Group II. Animals in Group V received CCl4 as in Group II, fish extract as in Group III, and rutin as in Group IV. Twenty-four hours post last treatment, all the animals were sacrificed by cervical dislocation after retro-orbital blood collection. CCl4-induced hepatotoxicity toxicity in toxic group rats was manifested as a decrease in body weight gain, an increase in ALT, AST, ALP, total and direct bilirubin, BUN, and creatinine in serum, a decrease in plasma total protein and albumin, decrease in antioxidant markers viz., SOD, CAT, GPx, and GSH and increase in lipid peroxidation marker, TBARS. Alterations in gross and histological architecture like fat deposition over liver surface, necrotic hepatocytes, cloudy swelling, fatty changes and hydropic degeneration were evidenced. Treatment with S. commersonnii fish extract and rutin either singly or in their combination, significantly improved the body weight gain and reverted the biochemical and histopathological alterations induced by CCl4. Though both fish extract and rutin individually exhibited significant hepatoprotective effect, it appeared that their co-administration produced better protective effect.ThesisItem Open Access STUDIES ON THE HEPATOPROTECTIVE EFFECT OF RASTRELLIGER KANAGURTHA FISH EXTRACT AND FLAVONOID MORIN IN RATS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2023-03) SRI HARSHINI VASAMSETTY; SRIVIDYA .G (MAJOR); RAVI KUMAR .P; NAVEEN SWAROOP .MHepatotoxicity is one of the major problem encountered in animals and humans, as the liver is the major target organ for the metabolism of various xenobiotics. Carbon tetrachloride (CCl4) is one of the most commonly used hepatotoxins in experimental studies to investigate liver injury associated with oxidative stress and free radical generation. The use of natural products, without causing much further stress, in preventing and treating liver diseases has gained wide acceptance. Fishes were reported to have higher amounts of essential amino acids, monounsaturated fatty acids, polyunsaturated fatty acids, and minerals. Rastrelliger kanagurta commonly called as Indian mackerel is widely distributed in the tropical Indo-Pacific region and is a rich source of omega-3 PUFAs which significantly regulate cell proliferation, fatty acid metabolism and in reducing inflammation and oxidative stress. Morin a bioflavonoid obtained from members of the Moraceae like Morus alba L (white mulberry), leaves of Psidium guajava, almond and other fruits possess various pharmacological properties such as antioxidant, anti-inflammatory, antineoplastic, antidiabetic, and neuroprotective properties. The major mechanism of hepatic damage produced by CCl4 is lipid peroxidation and oxidative damage by producing free radicals. As the flavonoid and fish extract possess antioxidant activity, they may reduce the hepatotoxicity produced by CCl4. Hence, the present study was designed to evaluate the hepatoprotective effect of Rastrelliger kanagurtha fish extract and morin alone and in combination in CCl4-induced hepatotoxic rats. Thirty Wistar strain male albino rats aged about 45 days were randomly assigned to 5 groups. Group I was maintained as control and received 1% DMSO orally for 3 weeks and olive oil I/P twice a week in the 2nd and 3rd week while groups II, III, IV, and V rats received carbon tetrachloride @ 1ml/kg body weight in olive oil (1:1) I/P twice a week in the 2nd and 3rd week. In addition, Group III & IV rats were administered with R. kanagurtha fish extract and morin at the dose of 300mg/Kg and 30mg/Kg body weight per oral daily for 3 weeks respectively, whereas Group V rats received combination of R. kanagurtha fish extract and morin per oral daily for 3 weeks. Twenty-four hours after the last day of treatment blood samples were collected for the estimation of biochemical parameters and liver samples were collected for gross and histopathological examination. Carbon tetrachloride-induced hepatotoxic rats showed a reduction in body weight, anemia and increase in transaminases. The natural defense mechanism of oxidative damage i.e., reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) were reduced and lipid peroxidation marker like TBARS increased significantly when compared to normal rats. Histopathological examination of the liver showed necrotic hepatocytes, ballooning of hepatocytes and centrilobular infiltration of mononuclear cells in sinusoidal spaces were evidenced in the CCl4 treated rats. Treatment with R. kanagurtha fish extract, morin, and their co-administration along with CCl4 led to a reversal in the hepatic biomarkers and a significant reduction in the MDA levels by 67%, 65% and 76% respectively. This trend was in agreement with improved body weight gain, improved antioxidant markers and less severe histoarchitectural changes in the liver. Based on the results it was concluded that the co administration of R. kanagurtha fish extract and morin may have a significant potential and synergistic effect on CCl4-induced hepatotoxicity than their individual treatments.ThesisItem Open Access STUDIES ON THE HEPATOPROTECTIVE EFFECT OF SARDINELLA LONGICEPS FISH EXTRACT AND FLAVONOID QUERCETIN IN RATS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2023-02) SIMRAN KOUR, S; SRIVIDYA, G(MAJOR); RAVI KUMAR, P; SUDHAKAR, KIn recent years there has been an increased research interest to develop therapeutic agents from natural sources. Fish serve as natural sources of medicines for cardiac disorders due to their richness in polyunsaturated fatty acids. Exploring the therapeutic potential of plant sources such as volatile oils, herbs phytochemicals was studied but limited research reports are available on the therapeutic potential of the fish extract on liver damage. Hence, the present study was designed to determine the effect of Sardinella longiceps fish extract and flavonoid quercetin on liver damage produced by carbon tetrachloride in male albino Wistar rats. Fish possess high protein content and also omega fatty acids known to support good health. The Indian subcontinent harbors a rich fish biodiversity which markedly varies in their nutrient composition. The Indian Oil Sardine (Sardinella longiceps) is a species of ray-finned fish of the genus Sardinella and is the most common species that forms the mainstay of the pelagic fishes of India. It has the highest lipid value (38%) and PUFAs which significantly regulate cell proliferation, and fatty acid metabolism, and in reducing inflammation and oxidative stress. Quercetin (3, 5, 7, 3, 4-pentahydroxy flavon), is a flavonoid considered one of the most prominent dietary antioxidants and occurs in a glycosylated form in French beans, broccoli, apples, and especially in onions. Male Wistar rats weighing 150-200 g, were randomly distributed into five groups, the first served as control and received 1% DMSO orally for 3 weeks and olive oil i.p. twice a week in the 2nd and 3rd week while groups II, III, IV, and V rats received carbon tetrachloride @ 1ml/kg body weight in olive oil (1:1) i.p. twice a week in the 2nd and 3rd week. The third and fourth groups were supplemented with Sardinella longiceps extract @ 300mg/Kg body weight and quercetin @ 30mg/Kg body weight orally daily for 3 weeks, respectively. Rats of group five were exposed to coadministration of Sardinella longiceps extract @ 300mg/Kg body weight and quercetin @ 30mg/Kg body weight orally daily for 3 weeks. Twenty-four hours after the last day of treatment blood samples were collected for estimation of indicators of hepatic damage and liver samples were collected for gross and histopathological examination. Administration of CCl4 for 21 days produced liver injury that was evident from elevated levels of serum enzymes such as ALT, AST, ALP, bilirubin, BUN, creatinine and histopathological alterations such as vacuolar degeneration of hepatocytes, and infiltration of inflammatory cells in sinusoidal space. The CCl4 treatment caused significant (p<0.01) changes in the cellular redox status as reflected by about a 79% decrease in the level of GSH, a marked decrease GPx, SOD, and catalase levels, and an increase in MDA in the liver. Sardinella longiceps extract (300mg/kg orally, daily ) markedly prevented the rise in serum enzyme levels elicited by CCl4. The histopathological alterations were also improved. Remarkably, there was a reversal of the changes to some extent in the levels of GSH and MDA, in animals receiving quercetin. The group receiving the coadministration of Sardinella longiceps extract and quercetin showed better protection against CCl4 and this was evident with improved body weight gain, decreased serum enzyme levels, and less severe histoarchitectural changes in the liver. In conclusion, the present study provided new insights into the use of fish as PUFA supplementation and the combination treatment was the most promising hepato protective formula among the different treatments.ThesisItem Open Access EVALUATION OF ANTIBACTERIAL ACTIVITY OF ACETONE EXTRACT OF Boswellia ovalifoliolata (INDIAN FRANKINCENSE) RESIN AND ITS SYNERGY WITH OTHER SELECTED PHYTOCHEMICALS AGAINST MASTITIS CAUSING ORGANISMS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2022-08) DIVYA SRI, TAPPITLA; BHARAVI, K(MAJOR); SRINIVASA RAO, G; ASWANI KUMAR, KMastitis is one of the major economic burden for the dairy industry and also has got huge public health significance. Conventional antibiotics comprise the most common therapeutic approach in mastitis, however their non-prudent use led to the development of resistant bacterial species. Steadily increasing rate of bacterial resistance to existing drugs necessitates the search of new antimicrobial agents to combat this problem. As an alternative to antimicrobial agent, phytochemicals offer effective and economical alternatives not only to treat infections but also counter bacterial resistane. Use of medicinal plant drugs does not carry the disadvantage of resistance and has the additional property of immunomodulation. The present study investigates the antibacterial activity and biofilm inhibition potential of acetone extract of Boswellia ovalifoliolata resin, its combined activity with commercially available antibiotics enrofloxacin and amoxycillin and with phytochemicals eugenol, thymol and carvacrol against standard MTCC cultures and clinical isolates Escherichia coli and Staphylococcus aureus from bovine mastitis. Preliminary phytochemical analysis of Boswellia ovalifoliolata resin extract (BORE) revealed the presence of carbohydrates, reducing sugars, saponins, triterpenes, phenols, flavonoids, proteins, and alkaloids. Antioxidant activity assessed by phosphomolybdenum assay with 0.1 mg/mL of BORE, eugenol, thymol, enrofloxacin, and 10 mg/mL of carvacrol which was equivalent to 63.1±0.89, 74.8±0.56, 53.80±0.32, 33.60±0.31 and 106±1.21 μg/mL of ascorbic acid respectively. Antibacterial activity assessed by microdilution assay and minimum inhibitory concentration values of enrofloxacin, BORE, eugenol, thymol and carvacrol against E. coli. were 0.078 μg/mL, 2.5 mg/mL, 1.562 mg/mL, 1.562 mg/mL and 1.562 mg/mL, respectively. MIC values of amoxycillin, BORE, eugenol, thymol and carvacrol against S. aureus were 0.3125 μg/mL, 0.3125 mg/mL, 3.125 mg/mL, 1.562 mg/mL and 0.390 mg/mL respectively for both MTCC cultures and clinical isolates. Synergistic antibacterial activity of BORE assessed by microdilution checkerboard assay in combination with enrofloxacin, amoxycillin, eugenol, thymol and carvacrol exhibited additive action against both E. coli and S. aureus. All bacterial isolates used are proved to be good biofilm producers after 24 hr of bacterial growth. Individually, BORE leads to a considerable biofilm reduction, but when tested in combination with antibiotics enrofloxacin, amoxycillin, eugenol, thymol and carvacrol are most effective in inhibition of biofilm. In conclusion, BORE contains various bioactive phytochemicals, exhibited potent antioxidant and considerable antimicrobial activity with MIC 0.3125 mg/mL, 2.5 against both gram-positive and gram-negative microorganisms, showed additive action in combination with enrofloxacin, amoxycillin, and phytochemicals eugenol, thymol and carvacrol and exhibited considerable biofilm inhibition at MIC values, alone and various combinations against bovine mastitis causing organisms E. coli and S. aureus.ThesisItem Open Access ROLE OF NARINGENIN ON THE ANTIBACTERIAL ACTIVITY OF SELECTED β-LACTAM ANTIBIOTICS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2022-03) RAJENDRA CHOWDARY, M; SRIVIDYA, G (MAJOR); RAVI KUMAR, P; SUDHAKAR, KBeta-lactam antibiotics are one of the most commonly used antibacterial agents against different bacterial infections in livestock. Among them ampicillin and amoxicillin are most widely used by the clinicians in both human and veterinary practice. However, their indiscriminate usage resulted in their reduced efficacy and development of resistance by microorganisms. To combat the resistance and restore the antibacterial efficacy of the antibiotics, their combination with non antibiotic drugs like phytochemicals is being considered actively.In the present study the drug interaction between selected β-lactam antibiotics (ampicillin and amoxicillin) and phytochemical naringenin was evaluated against Escherichia coli and Staphylococcus aureus invitro using micro broth dilution method, crystal violet assay and checker board method. MIC, MBC, FIC, and biofilm inhibition values were used as indicators to determine the antibacterial activity. MIC values of ampicillin, amoxicillin and naringenin against Escherichia coli were 6.25μg/ml, 12.5μg/ml and 2.5mg/ml respectively. The minimum inhibitory concentration values of ampicillin, amoxicillin and naringenin against Staphylococcus aureus were 0.312μg/ml and 0.312μg/ml and 0.625mg/ml respectively. The MIC values of ampicillin and amoxicillin were reduced by 2-fold in the presence of naringenin. The MBC values of ampicillin were significantly reduced from 25μg/ml to 12.5μg/ml in presence of naringenin against Escherichia coli and from 1.25μg/ml to 0.625 μg/ml against Staphylococcus aureus. FIC value of combination of ampicillin with naringenin against Escherichia coli and Staphylococcus aureus was 1 indicating the additive effect of the combination. The MBC values of amoxicillin were significantly reduced from 50μg/ml to 25μg/ml in presence of naringenin against Escherichia coli and from 1.25μg/ml to 0.625 μg/ml against Staphylococcus aureus. FIC value of combination of amoxicillin with naringenin against Escherichia coli and Staphylococcus aureus was 1 indicating the additive effect of the combination. Ampicillin, amoxicillin and naringenin alone inhibited the Escherichia coli biofilm formation by 63.5±2.25, 65.1±2.32 and 46.6±5.32 % respectively and combination of ampicillin with naringenin and amoxicillin with naringenin inhibited the biofilm formation by 44.3±2.89 and 54.4±2.68 % respectively. xvi Ampicillin, amoxicillin and naringenin alone inhibited the Staphylococcus aureus biofilm formation by 59.3±2.28, 59.7±2.41 and 33.02±6.8 % respectively and combination of ampicillin with naringenin and amoxicillin with naringenin inhibited the biofilm formation by 32.65±3.49 and 45.9±2.31% respectively. Based on the above results, it can be concluded that naringenin produced positive drug interaction with ampicillin and amoxicillin by reduction in the MIC and MBC values significantly. Further, in-vivo studies are required to establish antibacterial mechanism and biofilm inhibition mechanisms of naringenin to develop it into a therapeutic agent against infectious diseases.ThesisItem Open Access EVALUATION OF ANTIBACTERIAL ACTIVITY OF ACETONE EXTRACT OF Boswellia serrata RESIN AND ITS SYNERGY WITH OTHER SELECTED PHYTOCHEMICALS AGAINST BOVINE MASTITIS CAUSING ORGANISMS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2022-03) MOUNIKA, DEVETY; BHARAVI, K (MAJOR); RAVI KUMAR, P; ASWANI KUMAR, KMastitis in dairy cattle is a highly prevalent infectious disease, causing considerable economic loss worldwide which alter milk quality, reduce milk yield, and increase cost of production resulting in great economic losses to the dairy industry. The conventional antimicrobial agents have been the mainstay of mastitis therapy over the decades and these drugs have potential high cure rate, when the treatment is well‑targeted. However, use of antibiotics is associated with cost, the possibility of development of acquired drug resistance, residues in the milk. Steadily increasing bacterial resistance to existing drugs is a serious problem, and therefore there is a dire need to search for new classes of antibacterial substances, especially from natural sources. Unlike synthetic drugs, the antimicrobials of plant origin are not associated with side effects and have a great therapeutic potential to heal many infectious diseases. As an alternative to antimicrobial agents, phytochemicals offer effective and economical alternatives not only to treat infection but also to counter bacterial resistance. Use of medicinal plant drugs does not carry the disadvantage of resistance and has the additional property of immunomodulation. In the present study Boswellia serrata resin extract was evaluated for the presence of various phytochemicals (qualitative), antioxidant potential by phosphomolybdenum assay, antibacterial activity by microdilution assay and antibacterial activity in ccombination with antibiotics enrofloxacin and amoxycillin, phytochemicals such as thymol, xanthone and caffeic acid and plant extracts cashew nutshell extract, Ficus extract by checkerboard and biofilm inhibition by crystal violet assay. The preliminary phytochemical analysis revealed the presence of carbohydrates, saponins, triterpenes, phenols and flavonoids. The antibacterial activity assessed by microdilution assay and minimum inhibitory concentration values of enrofloxacin, amoxycillin, Boswellia serrata resin extract, cashew nutshell extract, Ficus extract, thymol, xanthone and caffeic acid against Escherichia coli and Staphylococcus aureus were determined. Combined antibacterial activity of Boswellia serrata resin extract assessed by microdilution checkerboard assay in combination with enrofloxacin, amoxicillin, aerial roots of Ficus extract, cashew nutshell extract and thymol exhibited additive action against both E.coli and S. aureus, synergistic and additive action in combination with xanthone against E.coli and S. aureus respectively, whereas indifference and additive effect in combination with caffeic acid against E.coli and S. aureus. All bacterial isolates used are proved to be good biofilm producers after 24hr of bacterial growth. Individually, Boswellia serrata resin extract leads to a considerable biofilm reduction, but when tested in combination with antibiotics enrofloxacin, amoxicillin and xanthone are most effective in inhibition of biofilm. In conclusion Boswellia serrata resin extract was effective against gram positive organism Staphylococcus aureus with lower MIC and biofilm inhibition compared to gram negative organisms Escherichia coli because of difference in the cell wall structure. Boswellia serrata resin extract showed additive effect and indifference in combination with antibiotics and phytochemicals and has no antagonist activity. Boswellia serrata resin extract in combination with xanthone showed synergy against Escherichia coli with maximum percent of biofilm inhibition.ThesisItem Open Access EVALUATION OF ANTIBACTERIAL ACTIVITY OF METHANOL EXTRACT OF TENDER PROP ROOTS OF Ficus benghalensis AND ITS SYNERGY WITH OTHER SELECTED PHYTOCHEMICALS AGAINST MASTITIS CAUSING ORGANISMS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2022-03) MONI THERESA, TSAPPIDI; BHARAVI, K (MAJOR); SRINIVASA RAO, G; ASWANI KUMAR, KMastitis caused by Escherichia coli and Staphylococcus aureus became a major health problem in bovines. Conventional antibiotics comprise the most common therapeutic approach in mastitis. But due to their indiscriminate use and high rate of resistant bacterial species development, the therapeutic alternatives to treat multidrug-resistant microorganisms are rapidly dwindling. The present study investigates the antibacterial activity and biofilm inhibition potential of methanol extract of Ficus benghalensis, its combined activity with commercially available antibiotics Enrofloxacin and Amoxycillin, with herbs Boswellia serrata resin extract (BS), Cashew nut shell extract (CSE), and with phytochemicals Xanthone (XA) and Caffeic acid (CA) against standard MTCC cultures and clinical isolates Escherichia coli and Staphylococcus aureus from bovine mastitis. Preliminary phytochemical analysis of Ficus benghalensis aerial root extract (FBARE) revealed the presence of reducing sugars, saponins, triterpenes, phenols, flavonoids, proteins, and alkaloids. Antioxidant activity assessed by Phosphomolybdenum assay with 0.1 mg/mL of FBARE, Cashew nut shell extract, Boswellia serrata resin extract, Enrofloxacin, Caffeic acid, and 10 mg/mL of Xanthone which was equivalent to 54.67±0.146, 63.6±0.64, 44.82±085, 33.605±0.31, 126.14±0.62, and 162±1.81 μg/mL of ascorbic acid respectively. Antibacterial activity assessed by microdilution assay and minimum inhibitory concentration values of Enrofloxacin, FBARE, cashew nut shell extract, B. serrata resin extract xanthone and Caffeic acid against E. coli. were 0.039 μg/mL, 2.5 mg/mL, 2.5 mg/mL, 5 mg/mL, 12.5 mg/mL and 6.25 mg/mL respectively. MIC values of Amoxycillin, FBARE, cashew nut shell extract, B. serrata resin extract, Xanthone and Caffeic acid against S. aureus were 0.3 μg/mL, 2.5 mg/mL ,0.078 mg/mL, 1.25 mg/mL, 25 mg/mL and 3.125 mg/mL respectively for both MTCC cultures and clinical isolates. Synergistic antibacterial activity of FBARE assessed by microdilution checkerboard assay in combination with Enrofloxacin, Amoxycillin, B. serrata resin extract exhibited additive action against both E coli and S. aureus, synergistic and additive action in combination with xanthone against E. coli and S. aureus respectively, whereas indifference and additive effect with CA against E. coli and S. aureus. All bacterial isolates used are proved to be good biofilm producers after 24 hr of bacterial growth. Individually, FBARE leads to a considerable biofilm reduction, but when tested in combination with antibiotics Enrofloxacin, Amoxycillin, CSE and Bos serrata are most effective in inhibition of biofilm. In conclusion, FBARE contains various bioactive phytochemicals, exhibited potent antioxidant and considerable antimicrobial activity with MIC 2.5 mg/mL against both gram-positive and gram-negative microorganisms, showed additive action when screened in combination with Enrofloxacin, Amoxycillin, CSE, Bos serrata resin extract, and Phytochemicals CA and XA and exhibited considerable biofilm inhibition at MIC values in alone and various combinations against bovine mastitis causing organisms E.coli and S. aureus.
