MICRO Aseptic Technique answer sheet (1)

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1 Diamond Simmons BIO 2104 March 11, 2024 Aseptic Technique Answer sheet Pre-Laboratory Questions 1. Is sterilization the same as disinfection? Explain your answer. - No, they are not the same. Sterilization is the killing of all microbes and disinfecting is the reducing and or eliminating harmful microbes from surfaces. 2. Why is aseptic technique important? - This technique protects patients from bacterial and viral infections as well as protection from microbes that could be harmful. This technique keeps the equipment used during surgical procedures clean so that no contamination occurs. 3. What are signs of microbial growth in a liquid medium, such as broth? On a solid medium, such as nutrient agar? - In a liquid medium cloudiness, gas bubbles, sediment at the top or bottom of the tube, and/ or color changes are signs of microbial growth. - In a solid medium, you would see texture changes, change in color and/or transparency. Observations Take a picture and insert it in the appropriate section below. For best results, avoid the use of flash. Do not remove the lid of the Petri dish or tube. Tilt the plates or tubes at slight angle: ©2016 Carolina Biological Supply Company

2 ACTIVITY 2: Aseptic Transfer from Broth Culture to Broth Activity 2, Broth to Broth Transfer, 48 hours incubation Activity 2, Broth to Broth Transfer, 72 hours incubation ©2016 Carolina Biological Supply Company

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4 ACTIVITY 4: Aseptic Transfer from Broth Culture to Plate Activity 4, Broth to Plate, 48 hours incubation Activity 4, Broth to Plate, 72 hours incubatio Data Table Activity Describe growth (turbidity or sediment if in a tube, colony characteristics if on a plate) Color of growth Amount of growth (Estimate coverage if on a plate, or state how turbid if a broth – i.e., could you read a paper through it?) Activity 2 48 hours Broth to Broth: Turbid Clear White The medium went from clear to a little cloudiness. Can hardly read through it. Activity 2 72 hours Broth to Broth: very Turbid Cloudy Very cloudy, cannot read through it Activity 3 48 hours Broth to Slant Clear Film visible, very light colored Activity 3 72 hours Broth to Slant: can see small white balls Light white Light liquid visible and colonies growing on the agar. ©2016 Carolina Biological Supply Company

5 Activity 4 48 hours Broth to Plate: Colony Forming White with small clusters It can be seen where the bacteria spread, and new colonies are forming. White in color. Activity 4 72 hours Broth to Plate larger colony groups Some red in center of clusters More colonies have formed, reddish balls can be seen in the center. Post-lab Questions 1. Should you see signs of microbial growth in a sterile medium if you allow it to incubate for several days? Explain your answer. - No, do to this being a sterile medium it has been sterilized and if there were any growth that would indicate contamination. 2. Do you think there were any deficiencies in your aseptic technique, and if so, can you identify them? Explain your answer. If there were no deficiencies in your aseptic technique, identify at least one potential way through which contamination could occur while working with cultures. -yes, I did forget to flame the tube before placing the lid back onto the tube. This may have let in microbes and caused contamination, affecting my outcome. 3. Which was most challenging for you in terms of aseptic technique: transfer from broth to broth, broth to slant, or broth to plate? Explain your answer. - For me it was the transfer from broth to plate. I was not sure if I let other microbes in with how high I lifted the lid as well as if I was pressing to hard or to light when smearing the broth. 4. Were there any discrepancies between what you saw in your experiment and what you expected to see based on the lab manual and your readings in this section? - I expected to see a lot more growth, when I did not see that much growth in the beginning, I wondered if I had messed up the experiment, but eventually growth occurred. ©2016 Carolina Biological Supply Company

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