One gram of liver was sliced into pieces, then 10ml of homogenization buffer was added to it. The mixture of the liver and homogenization buffer was then placed into a homogenizer to make the liquid slurry. Once the slurry was made it was placed into a 50ml falcon tube to then be placed in the centrifuge. The slurry was centrifuged for 2 minutes at 2000rpm. Once the spin in the centrifuge was complete, the slurry had separated, the most dense particles to the bottom of the tube forming sediment and the lighter (liver succinate dehydrogenase enzyme) also known as the supernatant. The supernatant was extracted from the falcon tube and placed into a test tube. The tube was then kept at a low temperature, (in an ice bath) until it was required for use.
pneumoniae) leaving me with Sa, Se, Ef, P.pyo, and Ml. Following my dichotomous key, the next test I decided to perform was a catalase test in order to determine if the enzyme catalase is produced by the bacteria. The sample of the bacteria I took did bubble, this indicates that the bacteria does produce the catalase enzyme as well as requiring oxygen to respire. Based on the results of the catalase test there were three possible microbes I could have Ml, Sa, and Se. For my next test I decided to use an MSA plate to determine if my unknown microbe could or could not grow in the presence of the mannitol salt, along with testing to see if it would or would not be able to ferment the mannitol salt. After allowing the bacteria to grow, I noted that the plate remained the same pinkish color it originally was however, I did several white colonies growing on the plate. The pink color that remained shows that the organism is unable to ferment the mannitol salt thus the acidic byproduct that turns the medium yellow will not be produced. Therefore the organism cannot be Sa since that
The sample of microorganisms in the air were tested with two different types of plates, Sabouraud’s plate and agar plate. Both the Sabouraud’s plate and nutrient agar plate were exposed to the same matter, the air , however, two plates showed different results of bacteria and mold. The Sabouraud’s plate contained 120 mold, but no bacteria, while the nutrient agar plate contained 5 mold, and 92 bacteria. The number of fungal organisms found on the Sabouraud’s plate was way more than the nutrient agar plate. The differences of the colony count is because of the environment of the two plates. The Sabouraud’s plate is acidic with the pH of 4, while the nutrient agar plate has a neutral pH of 7. Also, the Sabouraud’s plate has more carbohydrates and sugar compare to the nutrient agar plate. Accordingly, the environment of the Sabouraud’s plate is more suitable for the fungal organisms to grow than that of the nutrient agar plate. Also, the sample of hard cough contained the presence of the microorganisms. On the blood agar plate, there was partial hemolysis alpha and mold. The partial hemolysis alpha was small with yellow halo around, and mold was big, round, and fuzzy. There was no presence of bacteria that indicated a disease because there was no bacteria with golden yellow. Since the mouth is one of the suitable place where microorganisms can grow, kissing or sharing a
Pseudomonas aeruginosa produces a pigment called pyocyanin. The pigment is toxic to aerobic and denitrifying bacteria, while it is harmless to fermentative bacteria. The bacteria that cannot ferment are particularly susceptible to attack by pyocyanin.it is absolutely ineffective against p.aeruginosa and other pseudomonas species. Gram positive bacteria are easily killed by it than gram negative bacteria. Glucose is aptly suited for fermentation. When glucose is added to medium, facultative anaerobes can harness it for producing their energy source. Since the activity of pyocyanin toxin requires either robust active aerobic or anaerobic respiration, the toxin cannot inhibit the facultative anaerobes efficiently(Baron and Rowe 1981) when preparing selective media for pseudomonas aeruginosa, if glucose is added, it can harbour facultative anaerobes and anaerobes.
The cells were homogenized with five up-and-down strokes in a glass tissue homogenizer, and then centrifuged for 20 minutes at 10,000 × g.
Following experimentation approval and funding, the two main key dates are the initial experimental setup and the completion of experimentation, and along with it, the evaluation of results. Prior to the initial setup, materials must be obtained from online chemical suppliers, a process that can take many weeks. After all the materials are obtained, and proper training in the local laboratory environment is conducted, the cell plates will be set up and exposed to the candidate compounds. Over the course of the experiment, progress will be documented both qualitatively and quantitatively; TRAP assays and flow cytometry will be utilized to measure cell response, and phase contrast microscopy will be used to document cell plate transformations. Finally, following the end of experimentation results will be evaluated, thus concluding the
This lab demonstrated the active process of endocytosis, which is the movement of a substance into the cell in the form of a vesicle, a small, round sac. This allows the cell to bring extracellular solid material or particles into the cell. In phagocytosis, the particle outside of the cell binds to a receptor on the cell’s membrane which causes the membrane to form extensions called pseudopods. These extensions form a circle around the particle and pull it inward into the cell. Once inside, the vesicle that has formed, called a phagosome, will unite with a lysosome, an organelle that contains enzymes that will break down or digest the
In this experiment we wanted to observe the effects that different types of solutions have on living cells. The three types of solutions that we were looking for was: isotonic solution………
Phagocytes cannot interact with the encapsulated strains as antibodies are absent and both surfaces are electronegative, causing them to repel each other. More effectively, protease-resistant molecule produced by encapsulated strains inhibit or lyse polymorphonuclear leukocytes.
First, the α-2,6 sialylation can block the apoptosis mechanism40-41. Apoptosis is the mechanism of programmed cell death of multicellular organisms. The apoptosis process is induced by binding of cell galectins42-43. Galectins are a type of lectin and lectins are a class of proteins which can bind to specific proteins, via interacting with specific glycans. These galectins bind galactose of cell surface glycans to initiate the killing mechanism. It has been reported that α-2,6 sialyation significantly blocks galectin binding and inhibit apoptosis and this leads to the undesirable growth of cancer cells44-45. The α-2,6 sialyation also significantly blocks apoptosis pathways which use FAS46 and TNFR147 cell receptors.
Day 1: different agar plates were divided into four sections and the four microbes were streaked and incubated overnight ( at 37 °C)
The severity of the infection was varied according to many elements in particular the pathogenicity and virulence of the causative strain. The virulence of S. aureus is generally a multifactorial and due to the combined action of several virulence determinants, which augment tissue colonization, tissue damage, and hence disease (Bien et al.,, 2011), these proteins facilitate the bacterial attachment and colonization within the cellular and extracellular material of the host. Moreover, cellular proteins, protease, and toxins, which inhibit phagocytosis and thus interfere with the ability of the host to actively, hinder infection by a specific immune response. Hemolysin and other enzymes aid the bacterial population in the invasion of the host tissues (Lowy, 2000). S. aureus can bind a variety of proteins
As there is no buffer in which both SacI and BglII exhibit >50% activity, a sequential digest was necessary (Method 2.3.). The digest was successful and the resulting cells were then lysed.
It specifically catalyzing hydrolysis 1,4-beta-linkage between N-acetylmuramic acid and N-acetyl-D-glucosamine which are a very sturdy bond in the bacterial cell. This is where the lysozyme is targeted. In the mucous membrane, there is antimicrobial compounds such as lysozyme and secretory antibodies (IgA). Lysozyme can kill the bacteria but it less effective against the infection. Present of the lysozyme inhibit or kill the bacteria before they can colonized either in the mouth, eye or in other mucous secretion. Throughout this experiment, the lysozyme activity was determine using agarose lysoplate method. Using the sample collected the experiment was conducted within several days to see the clear zone, but the result obtained did not showed any clear zone even after added the coomassie brilliant blue stain. This may due to the plate bacteria inside the plate was dead when the sample was loadedin the well. There is possibility that bacteria was dead because the present of the bacteria in the plate cannot be seen and mybe the environment or the condition in the plate was not favourable for the bacteria to stay
Phosphate-Buffered Saline (PBS) was used to clean the excess media in the cell and to maintain the osmolarity of the cells. In the cell and tissue culture, PBS was needed to remove the serum of media for trypsin to detach the cells from the flask . The salt ions in the PBS was isotonic to the cell because of equal amount of salt ions inside the cells. The cell will burst if the solution contain a few of salt ion and would be shrink if too much contain of salt ions. Therefore, the amount of salt ions and osmolarity in PBS must be correct to keep the cells in an isotonic state.