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    Gram Staining

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    mike peter
    The document describes a gram staining procedure performed on a water sample to differentiate between gram-positive and gram-negative bacteria. Gram staining works by trapping crystal violet dye within the thick peptidoglycan layer of gram-positive bacteria, making them appear purple or blue, while gram-negative bacteria wash out and take up the pink safranin counterstain. The results of analyzing the water sample showed a purple color, indicating the presence of gram-positive bacteria. As some gram-positive bacteria can cause disease, the conclusion recommends treating the groundwater before using it.

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    Gram Staining

    Uploaded by

    mike peter
    18 views3 pages
    The document describes a gram staining procedure performed on a water sample to differentiate between gram-positive and gram-negative bacteria. Gram staining works by trapping crystal violet dye within the thick peptidoglycan layer of gram-positive bacteria, making them appear purple or blue, while gram-negative bacteria wash out and take up the pink safranin counterstain. The results of analyzing the water sample showed a purple color, indicating the presence of gram-positive bacteria. As some gram-positive bacteria can cause disease, the conclusion recommends treating the groundwater before using it.

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    The document describes a gram staining procedure performed on a water sample to differentiate between gram-positive and gram-negative bacteria. Gram staining works by trapping crystal violet dye within the thick peptidoglycan layer of gram-positive bacteria, making them appear purple or blue, while gram-negative bacteria wash out and take up the pink safranin counterstain. The results of analyzing the water sample showed a purple color, indicating the presence of gram-positive bacteria. As some gram-positive bacteria can cause disease, the conclusion recommends treating the groundwater before using it.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    18 views3 pages

    Gram Staining

    Uploaded by

    mike peter
    The document describes a gram staining procedure performed on a water sample to differentiate between gram-positive and gram-negative bacteria. Gram staining works by trapping crystal violet dye within the thick peptidoglycan layer of gram-positive bacteria, making them appear purple or blue, while gram-negative bacteria wash out and take up the pink safranin counterstain. The results of analyzing the water sample showed a purple color, indicating the presence of gram-positive bacteria. As some gram-positive bacteria can cause disease, the conclusion recommends treating the groundwater before using it.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    of 3

    NOVEMBER 2021

    GRAM STAINING ON COLLECTED WATER SAMPLE


    .

    Objective of experiment
    This test differentiates the bacteria into Gram-Positive and Gram-Negative Bacteria, which helps in the
    classification and differentiation of microorganisms.

    Introduction:
    The two major groups of bacteria can be divided into gram-positive and gram-negative. The Gram stain
    technique is based on the differential structure of the cellular membranes and cell walls of the two groups.

    Gram-positive organisms contain a highly cross-linked layer of peptidoglycan that retains the primary dye,
    crystal violet (CV), following the application of the mordant, iodine (I). The iodine and crystal violet form a
    complex within the peptidoglycan. When decolorizer is applied to the cells, the CV-I complex remains
    within the cell, making it appear dark purple to blue.

    The gram-negative organisms do not contain a thick cross-linked layer of peptidoglycan. The peptidoglycan
    is loosely distributed between the inner cell and the outer cell membranes. Following the application of the
    crystal violet and iodine, the CV-I complexes are not trapped within the peptidoglycan. Application of the
    acid-alcohol decolorizer dehydrates the outer cellular membrane, leaving holes in the membrane and
    effectively washing or removing the CV-I complex from the cells. The cells appear colorless. To make the
    colorless cells visible, a secondary stain, safranin, is applied, leaving the gram-negative cells pink.
    PROCEDURE FOR GRAM STAINING:

    EQUIPMENT:

     Electric microscope
     Microscope slide
     Looper
     Gas burner
     Distilled water

    REAGENTS

     Crystal Violet, the primary stain


     Iodine, the mordant
     A decolorizer i.e alcohol (95%)
     Safranin, the counterstain

    Procedure:

    1. Prepare and sterilize the microscope slide.


    2. Sterilize the looper by heating on the burner
    3. Smear a loopful of sample broth on the slide.
    4. Heat fix the sample on the slide by passing it through the flame about 3 times
    5. Flood the smear with crystal violet, the primary stain, for 30 seconds.
    6. Gently rinse off the excess stain with distilled water.
    7. Flood the smear with Gram’s iodine, the mordant, for 1 minute.
    8. Gently rinse off the excess iodine with distilled water.
    9. Run the acid-alcohol decolorizer over the smear until the solution appears clear.
    10. Gently rinse with distilled water for 30 seconds.
    11. Flood the smear with safranin, the secondary or counterstain, for 30 seconds.
    12. Gently rinse off the excess stain with distilled water.
    13. Blot dry with bibulous paper.
    14. Place the slide on the microscope stand and view under 100X objective lens

    OBSERVATION
    A purple color was observed on the specimen
    DISSCUSSION

    The purple color is an indication of gram-positive bacteria. This is because Gram positive result
    in Blue/Purple Color and Gram-Negative result in Red/Pink Color

    Some examples of Gram-Positive: Streptococcus, Staphylococcus, Corynebacterium, Listeria,


    Bacillus, Clostridium, etc

    CONCLUSION/ RECOMMENDATIONS:

    Therefore, from results of this study, the groundwater in the study area can be regarded to
    contain a gram positive bacteria. Hence the water should be treated using appropriate methods
    before it is used for domestic purposes or as drinking water.

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