Characteristics Of A Gram Stain Essay

An unknown bacterium was handed out by Dr. Honer. The appropriate tests were prepared and applied. The first procedure that was done was the gram stain. Under a microscope, if the gram stain is purple, the bacterium is gram positive, if the stain is red, it is gram negative. The next test was the fermentation tests for glucose, sucrose and

The purpose of this lab was to identify an unknown bacteria culture using differential tests. The identification of the unknown culture was accomplished by identifying the bacteria based on its specific metabolic characteristics and morphology. It is suggested that culture 11 is a sample of Enterobacter aerogenes.

The Gram stain was used to determine if the bacteria was gram positive or negative. A negative test shows a pink color and a positive test is a purple color. When a bacterium is negative it is because it has an outer membrane and a thin layer of peptidoglycan that is much harder to stain. A positive bacterium has a thick layer of peptidoglycan and no outer membrane that can be penetrated by crystal violet.

I identified Citrobacter freundii, the gram negative rod, by running a series of tests. I began with the Phenol Red Lactose tests, which tests if the organism contains various enzymes that determine if it can ferment lactose. The broth turned yellow after it was incubated, indicating that the lactose was fermented to acid, and there was also gas present in the Durham tube. Since the Phenol Red Lactose Test was positive, I then ran the Phenol Red Sucrose test, which tests if the bacteria contain different enzymes that determine if sucrose can be fermented. After incubation, the broth was yellow, indicating that sugar was fermented to acid, and there was also gas present in the Durham tube. Next, I ran the Sulfide Production, Indole Formation, Motility test, but I was only testing for Hydrogen Sulfide Production to differentiate between the organisms Citrobacter freundii and Enterobacter aerogenes. This test detects if the organisms can metabolize sulfur into hydrogen sulfide, which is revealed by the formation of ferrous sulfide that causes blackening around the growth. The test also reveals if the organism can break tryptophan into indole or migrate away from initial stab area. After incubation, the agar slant was completely black, indicating that the organism produces hydrogen sulfide and is motile proving that it was Citrobacter

This means that the bacteria for this lawn have cells that are surrounded by a cell wall which includes a thin layer of peptidoglycan and surrounding that is an outer lipid membrane that does not contain the Gram strain. Yet at the same time while Kanamycin was the most effective the Ampicillin and Penicillin had reactions as well. The Ampicillin would kill both Gram-positive and Gram-negative so this could indicate that there are Gram-negative bacteria in the lawn, but the fact that the Penicillin worked as well was what was interesting. This means that this bacterial lawn contains both gram-positive and gram-negative bacteria. Gram-positive are bacteria with cell walls that contain a thick layer of peptiodoglycan that retains the Gram strain. For plate 2, trashcan lid, only the Kanamcyin worked which indicates that the only bacteria in this lawn are Gram-negative, but if that is true why didn't the Ampicillin work at all? Ampicillin works to kill both positive and negative grams, but there was no reaction this time. Could it be that it needs more

Escherichia coli cells would be pink following a gram stain. E. coli is a gram negative cell due to its thin peptidoglycan layer and its outer membrane structure. In the gram staining process, the outer membrane does not play a role in staining but the peptidoglycan

In the Voges-Proskauer test, I inoculated the tube with bacteria from my TSA plate and incubated the tube at 37 degrees Celsius for 3 days. After three days I placed some Barrits Reagent A and Barrits Reagent B in the test tube. The color change of red or pink indicates a positive reaction for acetoin which tells you that the organism is a butanediol fermentor.

This experiment was given to us to utilized previous knowledge learned throughout the semester to identify a gram negative unknown bacterium. We had to first learn the difference between a gram negative and a gram positive organism. We started off with doing gram stains to determine whether it was positive or negative. Based on the gram stains, a gram positive stains purple and a gram negative stains pink. A gram positive stains purple because the cell walls is made of a thick peptidoglycan layer and doesn’t

Triple Sugar Iron Agar test, there was a gas production seen in the media. The media was yellow slant and yellow butt indicating glucose, lactose and/or sucrose fermentation with acid accumulation in slant and butt. For sulfur reduction, it was negative since it did not turn black in color indicating no sulfur was reduced.

The purpose of this lab was to identify an unknown microorganism using lab techniques. The importance of identifying microorganisms is essential to the survival of humans, expansion of modern day medicine and improvement of quality of life. In 1884, Hans Christian Gram designed a differential staining technique to identify bacteria that would change the future of microbiology. He give rise to a staining process, known as the Gram stain to differentiate microorganisms into two groups between positive and negative gram staining microorganisms. The Gram stain is essential in a lab technique as it distinguishes the cells based on the physical properties of the individual cell walls, and is almost always the first test preformed to differentiate a microorganism. The identification of weather a microorganism is gram positive or negative can revel the bacteria’s virulence, cell wall structure, resistance to antibiotics, resistance to physical disruptions and so much more. In order to identify the unknown provided, unknown #27, the Gram stain was the first test preformed. After discovering that the unknown bacterium was indefinitely a gram positive microorganism, the vast possibilities were narrowed down. However, In order to more definitively identify the unknown, the next step was to preform biochemical tests. A biochemical test identifies metabolic

8. Discuss a possible mechanism of Gram Staining in terms of differences in structure and chemistry between the walls of gram-positive and gram-negative

The chart below shows the biochemical tests of the gram stain below. Test performed Purpose Materials used Observation Results Gram stain test Gram reaction to specimen Crystal violet, iodine, alcohol safranin Pink/purple colored Gram positive cocci/ gram negative rods After the gram stain showed the specimen as Gram positive cocci, and Gram negative rods, more tests are necessary. Test performed Results Enterotube II Gram negative rod: Citrobacter

Gram staining is a technique used to determine if the bacteria is Gram positive or Gram- negative. Gram staining procedure uses crystal violet stain, iodine moderator, alcohol decolorizer and safarin counter stain. In Gram- negative bacteria the primary stain will be washed out with the decolorizer and it will be stained with the counterstain. Whereas in Gram-positive bacteria the primary stain will not leave the cell wall. This difference comes from difference in the structure of the cell wall that retains the stain.

5) One final test was run on the sample and that was the EMB test. Results of this test showed little to no growth on the EMB plate while the color of the colony (although barely visible) remained the same color as the agar. This test is selective for gram negative bacteria but inhibits the growth of gram positives. It also is differential in that it can test whether a bacteria can ferment lactose. Due to the fact that there was virtually no growth (a very faint haze) and no color change of the agar, the sample can be said to be gram

Escherichia is a genus of aerobic gram-negative rod-shaped bacteria of the family Enterobacteriaceae that form acid and gas on many carbohydrates, such as dextrose and lactose, but not acetone, which include occasional pathogenic forms, including some strains of E. coli which are normally present in the human intestine as well as other forms which typically occur in soil and water (Webster). Escherichia coli is a gram-negative bacilli that rarely varies in shape and size and when stained often resemble safety pins because the ends of some bacilli stain more densely than does the middle; which is a characteristic called bipolar staining which is common in enteric gram-negative bacilli (ASM). Gram negative cells have a thin cell wall layer and will stain red to pink. The staining process is the same as Gram positive, requiring four steps: applying a primary stain, adding a mordant, then rapid decolorization and completing with a counter stain. Applying the alcohol for decolorization dissolves the outer membrane and leaves small holes in the thin peptidoglycan layer through which the crystal violet-iodine diffuse. The gram-negative bacteria is colorless after the decolorization; therefore adding safranin