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Importance of gram staining technique
Important of gram staining
Gram staining lab report Introduction
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ASSIGNMENT # 1: GENERAL PRINCIPLES OF MICROBIOLOGY
1. Describe the components of the bacterial cell envelope and the primary function of each.
The innermost layer of the bacteria cell envelope is the cell membrane. It is a typical phospholipid bilayer, as is found in animal cells. The primary function of the cell membrane is energy production and active transport. The next layer is the cell wall. It is made of N-acetyl glucosamine and N-acetyl muramic acid. Amino acids keep the cell wall together by forming cross-bridges. The primary function of the cell wall is to protect the bacteria from its environment. It also acts as an osmotic barrier and provides the shape of the bacteria. The outermost layer of the cell envelope is the outer membrane. The outer membrane contains lipopolysaccharides, which act as an endotoxin. However, only gram-negative bacteria cells contain the outer membrane. Also, found between the cell membrane and the cell wall is the periplasmic space. It is filled with periplasm, which is made of water and proteins. The proteins act in degradation, transport and motility.
(Source:http://www.microbiologytext.com/index.php?module=Book&func=displayarticle&art_id=59)
2. A 62 year old patient was diagnosed with shingles. This is termed a latent infection.
- What category of viruses causes latent infections?
DNA viruses cause latent infections. Retroviruses, such as HIV, also have the ability to become latent after reverse transcriptase enzyme creates DNA.
- There are some bacteria that also cause latent infections. With an example explain the difference between latent viruses and latent bacterial infections.
In latent viruses, such as herpes, the viral genome remains in the host cells, but not the infectious virons...
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8. Why is it important clinically to know the Gram stain and morphology of infecting bacteria?
Without completing the gram stain, it is very difficult to see the bacteria. The gram stain gives a few clues that narrow down what kind of bacteria this could be. First of all, the result of the gram stain (what colour it becomes), helps you identify whether it is a gram-negative or a gram-positive bacteria. The gram stain also allows you to see the shape of the bacteria. Both the staining characteristic and the shape help define which bacteria you are dealing with. Once this is known, it is much easier to treat. For example, there are very few gram-positive bacteria rods that infect humans. If this type of bacteria is found you have a very good idea about what the infection could be, and could use on of the antibiotics that work well on this type of bacteria.
Upon receiving the unknown Microorganism (M.O.) #16, I prepared a slide by cleaning and drying it. Then, using a bottle of water I placed sterile drop of water on the slide and used an inoculating loop, flame sterilized, I took a small sample of the unknown growth in my agar slant and smeared it onto the slide in a dime sized circle and then heat fixed it for ten minutes. After ten minutes had passed, I collected the ingredients needed to perform a gram stain. I got the primary stain, crystal violet, and flooded my smear for sixty seconds, and then rinsed the color off with water until the water ran clear. I then flooded the smear with the mordant, grams iodine, and let that sit on the slide for sixty seconds as well. I then rinsed the grams iodine off with water and applied alcohol to the smear to decolorize the cells; however I made sure not to over decolorize and only put enough drops on the smear till the purple ran clear. I then rinsed the slide with water and flooded the smear with safranin the counter stain and let it sit for sixty seconds and then rinsed the color off with water. I blo...
The first day an unknown sample was assigned to each group of students. The first test applied was a gram stain to test for gram positive or gram-negative bacteria. The morphology of the two types of bacteria was viewed under the microscope and recorded. Then the sample was put on agar plates using the quadrant streak method for isolation. There were three agar plates; one was incubated at room temperature, the second at 30 degrees Celsius, and the third at 37 degrees Celsius. By placing each plate at a different temperature optimal growth temperature can be predicted for both species of bacteria.
They can be found anywhere and identifying them becomes crucial to understanding their characteristics and their effects on other living things, especially humans. Biochemical testing helps us identify the microorganism present with great accuracy. The tests used in this experiment are rudimentary but are fundamental starting points for tests used in medical labs and helps students attain a better understanding of how tests are conducted in a real lab setting. The first step in this process is to use gram-staining technique to narrow down the unknown bacteria into one of the two big domains; gram-negative and gram-positive. Once the gram type is identified, biochemical tests are conducted to narrow down the specific bacterial species.
I was given unknown organism #14, in order to find out what organism I had, I had to perform several different biochemical tests to identify it. Starting with the Gram stain test, which is performed to differentiate Gram-positive and Gram-negative cells. After staining, when observed through the microscope Gram-positive cells are a purple color with thick peptidoglycan cell walls. Gram-negative cells are a pinkish/red color with thinner cell walls. (handout G. s.) My organism was observed to be pinkish rod shaped meaning it is Gram-negative bacteria.
The first step to the unknown is selecting an actual organism. The best way to select a culture is based on a high-quality distribution. Equally important, shaking up the broth tube facilitates in the distribution. Upon selection, a gram check for purity is performed. Step by step instructions for this procedure can be found in Benson’s, Microbiological Applications p. 99. Furthermore, an aseptic technique must be performed for this test and the entire tests following the unknown. The purpose of this test is to differentiate between gram positive and gram-negative bacteria. The key indicator of gram-positive bacteria is a purple stain and a pink stain for gram-negative bacteria. A slide is viewed with a microscope under oil immersion. Equally
If the oxidase test was to be negative then the bacteria would have to be S. flexnery. S. flexneri is a bacterium that causes diarrheal disease, and it’s a “facultative anaerobe belonging to the family Enterobacteriaceae” (“Shigella flexneri”, n.d). However, if the oxidase test came as positive then the bacteria would have to either P. aeruginosa or A. faecalis (“Microbiology 20 Biochemical Unknown, 2009). Then an indole test will have to be done. An indole test is to identify if the bacteria could produce the enzyme tryptophase (SIM Medium”, n.d). If the test came as positive then unknown bacteria 2 would be P. aeruginosa (“Microbiology 20 Biochemical Unknown, 2009). P. aeruginosa is a bacterium that is the number one leading infections in humans. The bacterium is Gram negative that can lead to “endocarditis, meningitis, etc. (Friedrich.M, Dec 5, 2016). In the other hand if the bacteria came as indole negative then bacteria 2 would be A faecalis. A faecalis is a Gram-negative, rod-shape bacterium with flagella, and that belongs to the family Alcaligenaceas”, and it’s an opportunistic pathogen that induces infection (“Alcaligenes faecalis”, n.d). All in all, skills were practices to determine two unknown
To perform this test we first did a Gram stain on our organism to determine if it was gram-positive or gram-negative. After this we performed a mixed Gram stain by incorporating our organism with a known bacteria that stained opposite of unknown. We were given the size of the known bacteria and performed a comparative analysis under the microscope to determine the size of our unknown. In my case the control was a gram-negative bacteria Escherichia coli.
The tools used by microbiologists are essential and advancements would not have been possible without the continued improvements of the microscope. In addition, the petri dish is another key factor and besides nature, it is the main source for cultured solid media that is viewed under the microscope. Additional tools include slides and the dyes that are used to determine changes in cells and microbes or pathogens that cause disease. Heat has also been identified as a major tool in killing deadly spores and the impacts of cold have identified the need for temperature controlled environments that kill viruses and prevent the decreased effect in medications. Heat and cold have both been identified as tools in treating various complications from
M proteins: M proteins are found on the surface of the organism and protect it against phagocytosis. The M proteins prevent the attachment of complement proteins to the cell. Complement proteins which are attached to the bacterium “tag” it for destruction by phagocytic cells, such as neutrophils and macrophages, in a process called opsonisation. By inhibiting this process, the M protein allows the group A streptococcus to survive longer...
Bacterial cells, like plant cells, are surrounded by a cell wall. However, bacterial cell walls are made up of polysaccharide chains linked to amino acids, while plant cell walls are made up of cellulose, which contains no amino acids. Many bacteria secrete a slimy capsule around the outside of the cell wall. The capsule provides additional protection for the cell. Many of the bacteria that cause diseases in animals are surrounded by a capsule. The capsule prevents the white blood cells and antibodies from destroying the invading bacterium. Inside the capsule and the cell wall is the cell membrane. In aerobic bacteria, the reactions of cellular respiration take place on fingerlike infoldings of the cell membrane. Ribosomes are scattered throughout the cytoplasm, and the DNA is generally found in the center of the cell. Many bacilli and spirilla have flagella, which are used for locomotion in water. A few types of bacteria that lack flagella move by gliding on a surface. However, the mechanism of this gliding motion is unknown. Most bacteria are aerobic, they require free oxygen to carry on cellular respiration. Some bacteria, called facultatibe anaerobes can live in either the presence or absence of free oxygen. They obtain energy either by aerobic respiration when oxygen is present or by fermentation when oxygen is absent. Still other bacteria cannot live in the presence of oxygen. These are called obligate anaerobes. Such bacteria obtain energy only fermentation. Through fermentation, different groups of bacteria produce a wide variety of organic compounds. Besides ethyl alcohol and lactic acid, bacterial fermentation can produce acetic acid, acetone, butyl alcohol, glycol, butyric acid, propionic acid, and methane, the main component of natural gas. Most bacteria are heterotrophic bacteria are either saprophytes or parasites. Saprophytes feed on the remains of dead plants and animals, and ordinarily do not cause disease. They release digestive enzymes onto the organic matter. The enzymes breakdown the large food molecules into smaller molecules, which are absorbed by the bacterial cells. Parasites live on or in living organisms, and may cause disease. A few types of bacteria are Autotrophic, they can synthesize the organic nutrients they require from inorganic substances. Autotrophic bacteria are either photosynthetic or Chemosynthetic. The photosynthetic bacteria contain chlorophyll that are different from the plant chlorophyll. In bacterial photosynthesis, hydrogen is obtained by the splitting of compounds other than water.
The infection control plays an important role for the prevention from bacteria and other microorganism that may affect the condition of the patient.
The term “microbiology” refers to the branch of study that deals with microorganisms. Microbiology is extremely important in today’s time for the crucial information that the study provides. Human’s have had a long and cruel history of disease and sickness, for example the bubonic plague, but microbiology gives scientists the ability to observe, study, and prevent sickness like the bubonic plague to ever happen again. At the center of microbiology lies the bacterial cell, one that differs from those of a plant or animal because it lacks a nucleus and membrane-bound organelles which, in turn are traded for pili, flagella, and in some cases a cell capsule. Bacteria that are capable of causing illness or disease are called pathogens, pathogens work by releasing toxins in the body or directly damaging the host’s cells. An article by Lise Wilkinson explains that the earliest categorizations of bacterial cells first occurred in the late eighteen-hundreds to the early nineteen-hundreds by scientists (at the time) O. Muller and C. Ehrenburg (Wilkinson, 2004). The observation and identification of unknown bacteria that emerge is crucial because these new bacteria might be pathogenic and cause illness so it is very important that the bacteria is identified as soon as possible in order to either prevent the upcoming illness or treat it. While the common person is unable to identify if they are carrying bacteria (which is very likely), specialized tests that are ran in a lab can identify different types of bacteria and can even help
The abnormal presence of bacterial growth can be inspected under a microscope. If the organism inspected is not the bacteria used in the experiment, it means that the growth of the bacterial culture investigated is absent. By using this method, contamination by foreign substances in the surrounding air can be ruled out and the results would be more accurate.