0.5 mL of AuNPs solution was added to the above mixture. The UV-Vis spectra were recorded with a time interval of 1 min in a scanning range of 200-600nm at ambient temperature (25±20C). Antimicrobial activity The agar disc diffusion method was employed for the determination of antimicrobial activity of the papaya leaf extract stabilized gold nanoparticles. The 0.1 ml of 108cfu/ml of different pathogenic bacteria suspension was spread on different plates nourished with LB media. Filter paper
Rose Bengal-(bis(aminoethyl)ethylene glycol) (2) from Rose Bengal disodium salt (1) The synthesis was done following procedure from . Rose Bengal Na+ salt (915 mg, 0.90 mmol) was dissolved in DMF (2ml) and DIPEA (0.312 ml, 1.80 mmol), HATU (308 mg, 0.81 mmol) were added. After activation for 15 min, the mixture was added to O-Bis-(aminoethyl)ethylene glycol trityl resin (309 mg, 0.31 mmol) preswollen in DMF for 2 hours. The coupling reaction wrapped in aluminum foil was allowed to proceed overnight on a nitrogen bubbler apparatus. The resulting red-burgundy coloured resin was filtered and washed well with DMF.
For ascorbate peroxidase assay extraction 241 buffer was supplemented with 1.0 mM ascorbic acid. The homogenate was centrifuged at 242 15,000×g for 15 min at 40C, and the supernatant was used as a crude enzyme extract. 243 Spectrophotometric determinations were performed using UV visible spectrophotometer 244 (UV-1700, Shimadju, Japan). 245 2.11.2. Estimation of superoxide dismutase (SOD) activity 246 SOD activity was estimated by its ability to catalyse NBT to formazan at 560nm 247 according to the method of Beyer and Fridovich (40).
* In the above RB, a calculated amount of 1.2 equivalent amount of PTSH was added during continous stirring. * To the RB a condenser was attached and it was put on refluxing for a time period of 18 hours at a temperature of 80 degrees in an oil bath. * Post refluxing, the condenser was removed and it was left undisturbed for 1 day and then cooled to a temperature of -20 degrees by keeping in the freezer for setting of hydrazone crystals. * Once the crystals were formed, the supernatant was decanted and the crystals were transferred to a beaker where they were washed with cold methanol and then dried. * These dried hydrazone crystals were used in the subsequent steps.
Absorbance versus Time Measurements: The absorbance was set to 0 Abs while the spectrometer was set to ʎmax (from Part A). In Part B, 1.00 ml of the solution was mixed with the Blue dye in the beaker and half-way covered with a cuvette. Concurrently, the Spectronic 20 was blanked with water. The processes detailed above were repeated, each at a time. The absorbance values were recorded for 11 minutes.
The natural agar consist of Yeast Extract, Tryptone , Lactose , Manniiol, Sodium Chloride , Dip otassium Hydrogen Phosphate ,Gelatin and Agar .Then , in a conical flask 14g of nutrient agar was mixed into 500 ml of distilled water and , The mixture was stirred and dissolved until most of the agar dissolve. By non-absorbent cotton wool plug the mouth of the flask was closed. By using the autoclave the agar was sterilized for 15 minutes.The cotton wool was removed. By the flame the mouth of the flask was heated before and after pouring the agar into the Petri dishes. And, the left hand the lid of the Petri dish was lift, just enough to enter the mouth of the flask and quickly was poured in agar (about 15 cm3).
2.2 Litmus Milk Reaction A milk-based, litmus broth tube is incubated and observed after 48 hours. Observations include lactose fermentation without gas as well as with gas, the reduction of litmus, casein protein coagulation and casein and protein hydrolysis. These characteristics were all determined based on the color of the solution and the production of a curd, the curds density and the production of a gas. To determine the density of the curd, the tube was slightly turned to see rather or not it was mobile or concentrated towards the bottom. 2.3 Carbohydrate Fermentation of Lactose, Sucrose and
We rinsed the penny in acetone and dried it with paper towel. Next, we determined the mass of the penny by placing it on a balance. The mass of the penny was 2.47 grams. Afterwards, we placed the penny in a beaker filled with 20 mL of 6 M HCl. In the end we put the beaker in the fume hood and allowed it to sit overnight.
5-aminotetrazole monohydrate: In a 250 ml round-bottom flask equipped with a condenser for refluxing (90 °C) and a magnetic stirring bar, 5.00 g (5.95 mmol) dicyandiamide (three times crystallized), 7.47 g (11.9 mmol) sodium azide and 11.00 g (17.8 mmol) boric acid and 100 ml of water is added and allowed to reflux for 24 hours, after the completion of the reaction, until the solution pH to about 2 to 3 as hydrochloric acid 37% is added (about 12 ml) Then the reaction mixture was cooled in a refrigerator for 18 hours and the white crystals formed. The mixture was filtered and washed three times with 10 ml of water and and dried in 60 °C for 5 hours and finally 45.8 g of product by it will be obtained. 5-Aminotetrazol monohydrate: Yield:,
The first of the four test was a Gram-stain. Once the experiment was completed the slide was then placed under a microscope where it was then categorized as Gram-negative bacilli. Its pink color when viewed through the microscope indicated that the test was Gram-negative and its rod shape indicated the morphology was bacilli. With these results the next test to be conducted was the citrate utilization test. A citrate tube was inoculated with P. aeruginosa and incubated at 37 degrees Celsius at 24 hours.