Monday, March 2, 2015

Gram Would Be Proud


     In our project this week, we began the process of determining the identity and makeup of our chosen soil microbe. In order to have the basic skills required to do this we used the Gram-staining technique to determine whether our microbe was Gram (-) or Gram (+) in three control groups of bacteria. The reason this information is useful to is because it gave us a better idea of what broader category of bacteria our microbe is identified in. But, it also plays a significant role in the medical field when determining how to treat an infection or sickness caused by bacteria. 

     Gram (+) bacteria have thicker cell walls made of peptidoglycan than absorb crystal violet stain very well and make the bacteria appear purple in color. Gram (-) bacteria have a thinner cell wall of peptidoglycan that is sandwiched between two layers of cell membrane. This structure allows Gram (-) bacteria to absorb the crystal violet initially, then after washing and counterstaining, the bacteria appear to be a pink color. The reason this is important in the treatment of bacterial infections is because Gram (-) bacteria are much harder to combat and develop a much higher resistance to antibiotics because of the cellular structure. The outer membrane of Gram (-) bacteria can prove useful when infecting hosts because it provides more protection from autoimmune bodies or antibiotics. This makes it more deadly and harder to treat in humans or animals. 

     In order to use this technique to identify our microbe, we first stained three other control microbes that resembled different Gram identities. Each one had already been previously determined so it could give us an idea of what each type of bacteria would look like before we tested our unknown soil microbe. 

                                             Gram-Stain Micrograph of control B. megaterium


     Then, we Gram-stained our unknown microbe and determined its shape and Gram-identity. Our group's microbe appeared to be of a diplococcous shape and stained purple meaning it was Gram (+). We recorded this result, but then did further testing to confirm our results from the Gram Stain. We used McConkey agar which is selective for Gram (-) bacteria because it contains a high concentration of salt which deters Gram (+) bacteria from growing. After allowing this agar to grow at 30 degrees Celsius for five days, we found that there was no growth of our unknown bacteria on the agar plate. This result was similar to the responsiveness of B. megaterium which is also a Gram (+) bacteria. So, the result of our McConkey agar confirmed our initial hypothesis that our unknown microbe is Gram (+). 

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