will be transferred to a clean glass slide and will be smeared and heat-fixed. The slide will be flooded with crystal violet for 1 minute and rinsing with distilled water. Next, the slide will be flooded with iodine solution for 1 minute, and rinsed with distilled water. It is then decolorized with alcohol and will be drained, and counterstained with safranin for 30 seconds. The slide will be rinsed and excess stain will be wiped out, and then air dry. Finally, the slide will be observed under microscope to determine the characteristic of the bacteria, and will be photographed using a cell phone camera.
Catalase test will be performed also, using protocol of Reiner (2013). A clean glass slide will be placed on a sterile petri dish. A single colony from a 18 to 25-hour culture will be inoculated and will be placed on a glass slide, because bacteria loses its catalase activity with age. A drop of 3% Hydrogen peroxide will be placed directly onto the organism and will be covered immediately. Results will be recorded and photographed.
3. Triple Sugar Iron Test
In triple sugar iron slant, the protocol of Acumedia Manufacturers Inc. (2010) will be used. A loopful of isolate will be inoculated. It will be stabbed into the slants to the bottom and will be streaked at the surface of the agar slants. The slants will be incubated for at least 18 to 24 hours at 37°C, and the results will be recorded afterwards.
4. Oxidation-Fermentation Test (OF Test)
In OF slants, a…
In order to obtain accurate numbers from the now grown-out Sordaria samples, a small procedure was followed. First, a sample of fungi was taken from the border of a WT and TT section in the Petri dish. A slide was prepared by placing one drop of water on the middle of it, the Sordaria sample on top, and a slide coverslip on top of the sample. The eraser of a pencil was used to lightly press on the coverslip in order to release the asci so that they can be seen. Under a microscope, a multitude of…
The ten Petri dishes that will exclusively not contain an antibiotic in bacteria culture serve as the control. There will be ten trials for the control and each level of IV. The experiment began by cleaning the work area and sterilizing it with 70% ethanol. Then trypticase soy agar (TSA) was poured into into six groups of 60 Petri dishes. Sixty different agar dishes were labeled based on the antibiotic used. Then the dishes were left to dry and solidify at room temperature for hour to rid of…
containing the phage is known as supernatant.The supernatant was filtered through a .22 micron filter in order to extract phages. Since bacteria and other particles tend to be bigger than .22 microns, they would not fit through the filter, thus allowing only phages through. Mycobacterium smegmatis was then added to the supernatant in the amount of .5 mL. It was allowed to incubate for 5 days. After incubating, 1.4 mL of the enriched filtrate was pipetted into a microcentrifuge tube. This was…
2 antimicrobial agents are against 2 different strains of bacteria within a cultured environment.
• Small Bottle of S.Aureus bacteria in Nutrient Broth
• Small Bottle of E.coli bacteria in Nutrient Broth
• 4 Nutrient Agar plates (2 for each bacteria)
• A P200 Gilson pipette
• A box of sterile P200 yellow tips
• Sterile spreaders
• Latex gloves
• Sterile filter disks
• Test samples of antimicrobial agents (Milton and Dettol)
• Sterile distilled water
Oral Microbiome 10: Reflection 3
One a daily basis the human mouth can come in contact with many different materials and organisms. “The human mouth is home to billions of individual microorganisms, including viruses, protozoa, fungi, archaea, and bacteria” (University of Minnesota Department of Biology Teaching and Learning, 2016, p. 21). In specific, during this lab we are looking at Streptococcus mutans (S. mutans) and Lactobacilli. In studies, it has been shown that yogurt has helped…
In this lab, we took four separate onion bulbs, labeled them A,B,C, and D, and put them in a beaker filled with water. We recorded the growth in number of roots and their average length for days one, two, and five. Then we put Bulbs B, C, and D in beakers with different amounts of caffeine and left Bulb A in water. We then recorded the number of roots and their average length on days two, three, and four. After that, we took and onion root form Bulbs A, B, C, and D, stained them, and looked for…
2.5. Morphological Identification/Evaluations of Entomopathogenic Fungi
The six isolates of fungal entomopathogens were provided by Department of Entomology, Penn State USA. Preliminary morphological identifications of fungal entomopathogens such as color of colony, mycelial characteristics and spores from all six isolates were done at microscopic level.
2.6. Molecular Characterizations of Entomopathogenic Fungi
Cultures of Metarhizium fungus were grown in nutrient broth (Bacto, Australia) in…
for proper aseptic technique.
a) The lid of the first agar plate was removed and the exposed agar was placed in the building on the laboratory rack for 3 hours. The lid was replaced and labelled as “air”. It was then incubated at room temperature.
b) A second Petri plate was divided in half using a marker. Two cotton sterile swab were moisten in sterile water. One of the swab was rubbed over one of the stool in the laboratory and the other swab was rubbed over the sink in the laboratory. Both…
growth compared to the other strain of yeast.
In this experiment, we used two different sugars, sucrose and glycerol, and individually placed them into two different strains of yeast. In this experiment each of the petri dishes contained 1/3 of 143 flo ↑, 140 flo ↓, and 138 WT. However, 138 WT was not inoculated because it was not being tested. For this experiment, we inoculated two kinds of sugars on a sterile cotton swab and then swabbed two petri dishes with sucrose and two petri…