Cup Plate Method | Cultivation Of Microorganism

loving cup Plate Method Cultivation Of MicroorganismAn Adduct formed by stirring (0.01 mole) of aromatic aldehyde with the 40% of NaHSO3. O-phenylenediamine (0.01 mole) was dissolved in 50 ml of warm Ethanol 80. The NaHSO3 adduct of the aldehyde is added slowly with constant stirring in the warm solution of O-phenylenediamine stirred for 20-30 min still solid return obtained, then added nose candy ml of Distilled pee and filtered . Now the product was recrystallised by using Ethanol.Step 2 Nicotinoyl Cloride0.1 mole of Nicotinic Acid was refluxed for 6 hrs with the 20 ml of Thionyl Chloride. After this the excess of Thionyl Chloride was distilled off and separated from the product and arid it.Step 30.01 mole of 2-phenylbenzimidazole solution in 100 ml Pyridine stirred for 8 hrs constantly with the 0.01 mole of Nicotinoyl Chloride ,then the water added 50 ml to get a solid product. The product was filtered, dried and recrystallised using Ethanol. evasionAIM AND OBJECTIVESMolecula r modification of a promising adopt rise is still a major line of approach for the discovery of impudent drug. Molecular modification involves substituting, elimination, or adding new moieties to a p arent lead chemical compound, there by making gradual changes in the physico-chemical properties of the parent compound and thus biological activity of the compound.It is clear from the literature review that a number of Benzimidazole derivatives are known for the, anti bacteriuml, antifungal and ant-inflammatory activities properties.The state studies were performed with the following objectives tax deduction of new series of 1,2-substituted benzimidazole derivatives.Characterization of newly synthesized compounds by spectra manners viz.infrared spectra (IR spectra), Nuclear charismatic resonance spectra (H NMR spectra) and (Mass spectra). viewing of the antibacterial and Antifungal of the newly synthesized compounds using respective(a) strains of bacteria and fungi by determini ng their MIC.Screening of anti-inflammatory action of Benzimidazole derivatives.Scope and Plan of workLiterature heap revealed that Benzimidazole nucleus is a part numerous class of reported molecules exhibiting versatile range of biological activities like antibacterial, antifungal, antiviral, anticancer, analgesic ,anti-inflammatory activity, antihyperlipidemic, antihistaminic, antiulcer, anti-arrhythmic , HIV-RT inhibitor. Considering the reported data round Benzimidazole nucleus we have tried to synthesize some Nicotinoyl derivatives of Benzimidazole. The Benzimidazole derivatives of all supra mentioned activities are mostly of 2-substituted type .The synthesis of 2-(substituted phenyl)-benzimidazolyl-1-pyridinyl-3-methanone was carried out(p) and screened for antibacterial, antifungal, and anti-inflammatory activity.The present work was divided in to three sectionsSynthesis of 1,2-substituted derivatives of Benzimidazole .Chemical motion-picture show of the synthesized com pounds.Biological evaluation of synthesized compounds.Pharmacological screening of the synthesized compounds.ANTIMICROBIAL SCREENINGAn antibiotic drug is a chemical compound that in high dilution hinders the egression and the natural selection of one or more species of microorganism.A drug is considered to have bacteriostatic or fungistatic activity when it inhibits the growth of bacteria or fungi respectively and antiseptic or fungicidal activity when it kills the bacteria or fungi. In vitro renders are used as screening procedure for new agents and for testing the cogency of individual isolates from infection to determine which of the available drug might be useful therapeutically.Important factors for antimicrobial activity are size of the inoculums, metabolic state of microorganism, pH, temperature, and duration of interaction, concentration of the inhibitor and presence of interfering substance.ANTIBACTERIAL exercise STUDIESLiterature survey reveals that the synthesis a nd evaluation of antibacterial activity of various 2-substituted benzimidazole derivatives. The development of resistant among various pathogenic microorganisms towards the antibiotics has increased the impetus for investigation new antimicrobial agent. When a compound are synthesized in the take to that one of them would be more effective than the existing one. The antimicrobial intensity of a compound can be evaluated by serial dilution method and cup plate method. Dilution susceptibility tests are used to determine the tokenish Inhibitory Concentration (MIC).MIC is the lowest concentration of a drug that inhibits the growth of a particular organism under specific condition. The sensitivity of a compound against a particular organism can be canvas by cup plate method.Initially the zone of forbidding method was carried out to evaluate the sensitivity of the organism were selected for determination of MIC.CUP PLATE method actingCultivation of MicroorganismThe following microorg anisms were used to study the antibacterial activity. vitamin B subtilis Gram positive bacteriaStaphylococcus aureous Gram positive bacteriaEscherichia coli Gram negative bacteriaSalmonella typhi Gram negative bacteria received Streptomycin (1000mcg)Solvent DMFAll the test compounds were tested at 250 g, vitamin D g , and 1000 g.Preparation of the averageComposition of nutrient agar-agar ordinaryBeef extract..10gPeptone..10gSodium chloride..5gAgar.20gPurified water1000mlpH 7.2 0.2The intermediate was prepared by dissolving the contract quantity of the dehydrated strength in purified water by modify on a water bath and were dispensed in 100 ml volume cone-shaped flasks. The conelike flasks were closed with cotton plugs and were sterilise by autoclaving at 121C (15 lb psig) for 15 minutes.The contents of the conical flasks were poured aseptically into sterile Petridishes are allowed to solidify. These sterilise Medias were used to subculture the bacterial culture.PROCE DUREEach Petridish was fill up to a sense of 4-5 mm with a nutrient agar medium that was previously inoculated with able inoculums of suitable test organism, and then allowed to solidify. The petridish were specially selected with compressed bottom and were place on level surface so as to ensure that the layer of medium is in uniform thickness. The petridishes were sterilized at 160-170C in hot air oven for 30 mins before use. low-pitched sterile borer of uniform size was placed approximately at 10 cm height, having an internal diameter of approximately 6-8 mm and make of aluminium (or) clean steel. Each plate was divided in to quaternity equal portions along the diameter. To each portion one cylindrical endocarp was made in medium with the help of sterile borer. Three cavities for test compounds and one cavity for the standard. The petridishes were incubated at 37C for 18 hours. diam of the zone of inhibition was measured and the average diameter for each smack was cal culated. The diameter obtained by the test sample was compared with that produced by standard Streptomycin.CUP PLATE mannerCultivation of MicroorganismThe following fungal strains were used to study the antibacterial activity.1. C.raphigera2. A.polytrichaStandard Ketocanazole (1000mcg)Solvent DMFAll the test compounds were tested at 250 g, 500 g , and 1000 g.Preparation of the mediumComposition of nutrient agar mediumSabraoud Dextrose broth..64gmDistilled water.1000mlpH..7.2 0.2The medium was prepared by dissolving the specified quantity of the dehydrated medium in purified water by heating on a water bath and were dispensed in 100 ml volume conical flasks. The conical flasks were closed with cotton plugs and were sterilized by autoclaving at 121C (15 lb psig) for 15 minutes.The contents of the conical flasks were poured aseptically into sterile Petridishes are allowed to solidify. These sterilized medias were used to subculture the fungal culture.ROCEDUREEach Petridish was filled to a depth of 4-5 mm with a nutrient agar medium that was previously inoculated with suitable inoculums of suitable test organism, and then allowed to solidify. The petridish were specially selected with flat bottom and were placed on level surface so as to ensure that the layer of medium is in uniform thickness. The petridishes were sterilized at 160-170C in hot air oven for 30 mins before use. Small sterile borer of uniform size was placed approximately at 10 cm height, having an internal diameter of approximately 6-8 mm and made of aluminium (or) stainless steel. Each plate was divided in to four equal portions along the diameter. To each portion one cylindrical cavity was made in medium with the help of sterile borer. Three cavities for test compounds and one cavity for the standard. The petridishes were incubated at 37C for 18 hours. Diameter of the zone of inhibition was measured and the average diameter for each sample was calculated. The diameter obtained by the test sample was compared with that produced by standard Ketocanazole.