Microbiology/Spring 2021 Name: Carolina Lab Activity Worksheet Incubation and Observations Experiment #1: Aseptic Techni

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Microbiology/Spring 2021 Name: Carolina Lab Activity Worksheet Incubation and Observations Experiment #1: Aseptic Techniques and Use of Media Incubate all cultures. After approximately 48 hours of incubation, examine your cultures. Do not remove the caps of your tubes or the lid of the Petri dish during any observation, because this may result in contamination. Take pictures of your cultures. For photographs of microbial cultures, avoid use of the flash, which will simply be reflected off the glass or plastic culture container. You may need to experiment with lighting conditions and camera angles to get the best pictures in your workspace. 1. Observe and photograph the cultures again after approximately 72 hours of incubation. Your inoculated media should exhibit increasing amounts of microbial growth throughout the incubation period. Record name of bacterial species used in this investigation, 3. 2. 4. Photo 1 Insert the photo(s) of your cultures taken after 48 hours of incubation.
Photo 2 Insert the photo(s) of your cultures taken after 72 hours of incubation. 1. 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.
Experiment #2: Streak Plate Method (Isolation) Photo I- Insert the photo of the plate taken at 48 hours Photo 2 - Insert the photo of the plate taken at 72 hours. 1. Did you obtain isolated colonies of both organisms? If so, explain how you know that you were successful. If not, explain why not.
Experiment #3: Simple and Differential staining. Bacterial Shapes Procedure 1 - Simple Staining Please complete the following data table Bacteria Procedure 2- Gram Staining Bacterial Are you able to observe at 10X objectives? If Yes what is color and shape of organism Are you able to observe at 10X objectives? If Yes what is color and shape of organism Are you able to observe at 40X objectives? If Yes what is color and shape of organism Are you able to observe at 40X objectives? If Yes what is color and shape of organism Gram Reaction (+ or -)
Discussion Questions 1. Why do the slides have to be heat fixed before staining? What do you think would happen if the slides were not heat fixed and then stained?
Observations Data Table 1 TSI (Triple Sugar Iron) Medium Fermentation Escherichia coli Species Staphylococcus epidermidis Micrococcus luteus Key to symbols: A-acid product accumulation from sugar fermentation, evidenced by color change to evidenced by color change to yellow NC no color change G=gas production from sugar fermentation evidenced by bubbles or fissures in the agar, or displacement of the agar Data Table 2: Oxidase Test Experiment #4: Biochemical Tests K alkaline product accumulation from protein catabolism, color change to red. H2S-hydrogen sulfide production evidenced by black precipitated in the agar; indicates sulfur reduction and fermentation Species Escherichia coli Reaction(s) observed Staphylococcus epidermidis Micrococcus luteus Color change (yes/no) and color observed
Data Table 3: Catalase Test Species Escherichia coli Staphylococcus epidermidis Micrococcus luteus Bubbles (yes/no)