Blog #7 :3/4/24 - 3/8/24

 Introduction 

This following week was the last week before spring break. This week I inoculated, gram-stain, and made some media. We did two chemical tests, an oxidase test and oxygen requirement test. I also pour plates for the first time 

Methods

I inoculated and gram-stained by the standard directions. I also made 100 ml of TGY 

  • deionized water -100ml

  • Agar- 1.50g 

  • Tryptone- .3g

  • Yeast Extraction- .3g

  • Dextrose- .1g 

pouring plates 

We take autoclaved TGY broth and heat it on the hot plate. After heating it let it slightly cool down, to test if it is the right temperature touch the cool-down media to your skin. If the media is ready, line up your empty plates closed at the edge of the table. To pour, slightly open the plate and pour into the media Just till the bottom is covered with media. Then leave the plate at room temperature to Let them solidify. 


Oxidase test

To perform the Oxidase test, you start by inoculating a plate with bacteria, in this case, D. sonorensis, and let it grow. Then in the fume hood add the reagent and observe the reaction. If there is a color change to blue/black, it's a positive result. If there is no color change it is a negative result. 


Oxygen requirement test

To perform the Oxygen requirement test you will need tubes of thioglycollate broth. For our experiment, we had four tubes of thioglycollate. Each two was inoculated in this sequence 

•Tube 1- bacillus subtilis

•Tube 2- E. Coli 

•Tube 3- D. sonorensis 

•Tube 4- nothing 


Results & Conclusion 

The result of the gram stain of caeni turned out wrong. Caeni is a gram-negative bacteria. My stain appears to be gram-positive. The inoculation was done without any problems and the growth was good. The result of the oxidase test was positive. This means that D. sonorensis produces cytochrome c oxidase enzymes. The result of the oxygen requirement test was that D. sonorensis is a microaerophilic. This means that D. sonorensis requires oxygen but at lower concentrations. 

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