Aseptic Technique answer sheet (1)

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Apr 3, 2024

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1 Aseptic Technique Answer sheet Pre-Laboratory Questions 1. Is sterilization the same as disinfection? Explain your answer. Sterilization is not the same as disinfection. Sterilization kills everything, including tough spores, while disinfection lowers germ levels to a safe point but doesn't kill every single germ. Sterilization is vital in medical places to wipe out all germs, while disinfection is about stopping germs from spreading on surfaces or objects. 2. Why is aseptic technique important? Aseptic techniques are necessary in medical and laboratory environments to prevent contamination and maintain sterility. It lessens the entry of harmful microorganisms, ensuring product quality, enhancing patient safety, and aiding in the prevention of illnesses. 3. What are signs of microbial growth in a liquid medium, such as broth? On a solid medium, such as nutrient agar? Increased turbidity, pellicle development, sediment formation, gas production, and pH variations are indicators of microbial growth in liquid media. Microbial growth on solid media is characterized by visible colonies that exhibit a range of sizes, colors, forms, and textures. Additionally, surface growth, hemolysis, and pigmentation are detected. 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: ACTIVITY 2: Aseptic Transfer from Broth Culture to Broth Activity 2, Broth to Broth Transfer, 48 hours incubation ©2016 Carolina Biological Supply Company

2 Activity 2, Broth to Broth Transfer, 72 hours incubation ACTIVITY 3: Aseptic Transfer from Broth Culture to Slant Activity 3, Broth to Slant, 48 hours incubation ©2016 Carolina Biological Supply Company

3 I used the wrong paper date, this is supposed to be 48 hours, but the picture is correct. Activity 3, Broth to Slant, 72 hours incubation I used the wrong paper date, this is supposed to be 72 hours, but the picture is correct. ©2016 Carolina Biological Supply Company

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5 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 turbidity clear Barely visible Activity 2 72 hours turbidity cloudy White fog like but see through Activity 3 48 hours Low turbidity Clear and streaky Clear but visible Activity 3 72 hours High turbidity White and cloudy Good coverage about, 80 percent Activity 4 48 hours colonies' growth following the inoculation with loop swabs. White and foggy Les than 20 % coverage Activity 4 72 hours Growth is showing following the streak created Smear appearance 45% coverage 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, you shouldn't see any signs of microbial development in sterile media after a few days of proper incubation. The presence of microbial proliferation indicates contamination because sterile medium is free of any living microorganisms. Contamination may result from incorrect handling of the medium, pathogen introduction during inoculation, or insufficient sterilization procedures. Thus, the microbial growth in an apparently sterile medium suggests a possible violation of aseptic technique or sterilization process, which might affect experiment results or lead to incorrect conclusions. 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. ©2016 Carolina Biological Supply Company

6 It was very scary to do this experiment, proper handling of the cultures as well as proper hygiene techniques was crucial. I feel as if I did not have deficiencies in my experiments. It was always very close. The changing of hands and making sure that one cap did not touch another was the most difficult for me. One very easy potential way through which contamination could occur is equipment contamination. If contaminated tools like pipettes, culture tubes, or media bottles are not handled or sterilized correctly, they might transfer germs into cultures. 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. The most challenging for me was broth to broth, only because transferring form liquid to liquid and at times your hand getting at times unstable can be difficult. This for me was because it was hard to not tip something over while flame sterilizing. If this happens cross contamination can lead to loss of the sample. 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 don’t think there were discrepancies between what I saw in my experiment, what could possibly be my own would be being a beginner which might influence challenges or discrepancies due to lack in technique execution. ©2016 Carolina Biological Supply Company

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