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10.4: MAC Media Procedure

  • Page ID
    131062
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    MacConkey Media Procedure  

     

    Name: _____________________________________________________ 

    Course Section: ______________________________________________

     

    The following activity will teach students how to inoculate an MAC agar plate. Post-inoculation, students will use their colony growth to determine if the bacteria are gram positive or gram negative and the bacterial rate of lactose fermentation. 

     

    Materials
    • 1 MAC Plate
    • 3 Stock Plates
      • Stock plates will contain 1 of the following bacteria: Escherichia coli, Serratia marcescens, or Staphylococcus epidermidis 
    • Sharpies
    • Parafilm
    • Inoculation Loops
    • Incinerator 

     

     

     

    Procedure

    Step 1: 

    • Get into groups of 2. 
    • Label the bottom of your nutrient agar plate with your initials, course section, and date.
    • Using a sharpie, split the bottom of the agar plate into 3 sections and label each section with a different bacteria. 
    clipboard_ec524cbdfebd2f1147697cea9055dde35.png
     
    Procedure

     

    Step 2: 

    • Heat your inoculation loop in the incinerator for at least 10 seconds until the wire is red/orange. 
    • Allow the loop to cool, but do not wave it around the air.
    • You can test if the loop is cool by touching it to an area of agar that does not have bacteria and listen for a "sizzle" burning sound. 

     

     

    Step 3: 

    • Once your loop is cool, aseptically collect 1 colony from the stock plate. You do not need to swipe or gather a large amount of bacteria. 
    • Partially remove the lid from your labeled sample plate, enough that you can reach the agar with the loop while still shielding the plate.
    • Gently touch the loop to the agar and move it back and forth in a zig-zag motion until one third of the plate has bacteria.
    • Be careful not to press too hard with the loop, you can cut or damage the agar. 
      • Make sure you are adding bacteria to the correctly labeled areas. 

     

     

    Step 4: 

    • Repeat Step 3 for the remaining bacterial species.
    • Make sure you are sterilizing your loop between bacterial species.
    • Do not drag bacteria into the different labeled areas of agar. This is not a quad streak. 
    • When you have inoculated all 3 bacterial species on 1 agar plate, seal your plate with parafilm and place it upside down in the 37°C incubator for 24 hours.

     

     

     

    Results

    In table below, record your observations for colonies grown on MAC media. 

    Results Table
    Bacterial Species Gram Positive or Negative Colony/Media Color  Rate of Lactose Fermentation

     

     

     

    Escherichia coli

     

     

     

     

     

     

     

     

       

     

     

     

     Serratia marcescens

     

     

     

     

     

     

     

     

       

     

     

     

    Staphylococcus epidermidis

     

     

     

     

     

     

     

     

       

     

    General Questions

    1. What adaptation allows enteric bacteria to survive in the human gut?

     

     

     

     

     

    2.How does MacConkey agar inhibit the growth of gram positive bacteria?

     

     

     

     

    3. Why does lactose fermentation change the color of the bacteria colonies and the MAC media?

     

     

     

     

    4. You grow an unknown species of bacteria on an MAC plate. The colonies turn a white color while the media turns yellow. What kind of lactose fermenter is your unknown species?

     

     

     

     

     

    5. P. aeruginosa bacteria target which group of people?

     

     

     

     

    6. Why is it important that a patient submit a clean urine sample when trying to determine if their UTI is caused by P. aeruginosa?

     

     

     

     

    7. You collect a urine sample from a patient with a UTI and culture the sample on an MAC plate. The bacteria grow in blue-green colored colonies, but there are no changes to the media color. Would you classify this UTI as being caused by P. aeruginosa? Explain your answer. 

     

     

     

     

     

     

     

     

     

     

     

     

     

     

    Attributions

    "Microbiology Labs II: Results, 12.6.1 MacConkey Agar" by Dr. Gary KaiserLibreTexts: Biology, Community College of Baltimore County, Catonsville Campus is licensed under CC BY 4.0

    "Microbiology Labs II: Pure Cultures, 3.4 Use of Specialized Media" by Dr. Gary KaiserLibreTexts: Biology, Community College of Baltimore County, Catonsville Campus is licensed under CC BY 4.0

    "Bio 221 Lab: Physiological Tests, 22.2 Selective and Differential Media-MacConkey, EMB, MSA" by Kelly BurkeLibreTexts: BiologyCollege of The Canyons is licensed under CC BY

     


    10.4: MAC Media Procedure is shared under a CC BY 4.0 license and was authored, remixed, and/or curated by LibreTexts.

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