Your Agar plate should now be sterilized, the process of sterilization should be finished. Procedure of the experiment: Prepare all the test tubes and place them in their respective test-tube holders. Put 10 millimeter of the 0.9 % saline solution in one test tube. Put 10 millimeter of the chicken broth in the other test tube. Put 1 capsule of the bioflorin into each of the test tubes and tap them until the capsule dissolves.
Materials and Methods To start with, the unknown bacteria # 710 broth had to be successfully isolated on an EMB and MAC agar plate. Using aseptic technique by sterilizing the wire loop with Bunsen Burner between inoculations and flaming the opening of the test tubes before inserting in the loop with the bacteria. The streaking technique used was to isolate the colonies on the agar plates. In addition, the streak plates had to be incubated in a upturned position for 24 hours in a hot temperature incubator at 37 degree Celsius. Bacteria need a favorable condition to grow in.
A spore-former would have green-pigmented endospore cells when looked at under the microscope. From the growth on the NSM, I smeared it aseptically to a wet slide. Slide was then left to be air-dried for about 10 minutes. It was important to heat fix the slide using a micro incinerator. The succeeding steps had to be taken with caution because the primary stain, malachite green, is toxic.
Four randomly selected Daphnia magna, for each trial, were removed from the provided colony for the bioactive compounds to be tested, and were transferred with a plastic wide-mouth pipette with approximately 10 mL of pond water to protect and ensure survival of the Daphnia. In order to acclimatize the Daphnia to laboratory conditions, they were then placed onto a petri dish on the Daphnia cooling chamber. The cooling chamber was located on the stereomicroscope platform and brought down the heart rate of the Daphnia to a range that was countable by the observer, since Daphnia heart rate at room temperature is too rapid. On the cooling chamber there were two petri dishes: one for the Daphnia that were going to be tested, and one with the Daphnia being tested on, to ensure constant consistent temperatures for each trial. To maintain a temperature conducive to the heart
Aseptic technique was initiated at the beginning of this experiment by cleaning the work surface with disinfected wipes. Personal protectives equipment was also worn. The material utilized in this experiment was: S. epidermidis culture broth, sterile cotton swab, streak plate, forceps in 70% alcohol, a lit tea light, and the three antibiotic disks (novobiocin, gentamicin, penicillin). The first step, I divided a plate into three quadrants and labelled them with the different antibiotic names. Using the lit tea light, like a bursen burner, I flamed the mouth of the S. epidermidis culture.
We had to go back into our notes and see what matched to what. Materials I used. Were a petri dish, scapula, liquids, burner, C-clamp, tongs, gas to start the flame, eight different types of powders, and the most importa martial was our goggles for safety. The petri dish was used to put the powder in for testing. The scapula was used for scooping up the powders.
Narrowing down the unknown microorganism to gram negative, this approach was helpful to take the next step, in some bacteria the cell wall is surrounded by cell enveloped called capsule, also some bacteria make capsule when faced in a harsh environment to protect them. A capsule stain was preform, the results were analyzed and observed. An additional procedure that was done, was the Fast Actin staining which helps to see if the bacteria contains Mycolic acid in their cell walls, which determines the structure and function of the cytoskeleton in living and fixed cells (Shah). As expected for both E.coli and K. Pnenumia the fast acting results were negative. For both E.coli and K. Pnenumia the Oxidase test was positive a reaction was obtained.
Adding 5% of ethanol to the slide enabled me to figure out how the Daphnia will act in that environment. One thing I had to make sure that it does not affect the experiment is the light in the microscope. I had to turn it off when the daphnia in on the slide to make sure that the heat doesn 't affect the Daphnia. Every two minutes I go to the microscope to check whether the Daphnia died or
Before starting the heating process, measure the weight of the crucible with its cover first and then tare the balance, and after that adding about 1 gram of the sample to the crucible with its cover, and then weigh it. Moreover, it is possible liberating harmful gases during the process of heating; therefore, being careful is important. The heating process ends when this sample changes the color to brown because water of hydration is removed to the sample. Additionally, give time to the small cool down and measure its weight. Next, transfer the sample to a 50 mL beaker and mixes with distilled water, which gets by rinsing the crucible with its cover in 8mL, so the solution is generated.
This information is found in (Table 1). After that we dissolved the Benzoic Acid we received three masses once the substance was dried (Table 2). During the drying there were some problems with the substance sticking to the stirring stick possibly causing loss of material. With the lowest mass of the Benzoic Acid (25.805g) was taken and subtracted from the evaporating dish mass to find the mass of the sample (Table 5) (Formula 1). After the Sodium Chloride was dissolved into