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    This document discusses the use of chemical agents for sterilization when physical sterilization methods are not suitable. It explains that chemical sterilization can use liquid or gaseous agents to kill microorganisms. Gaseous sterilization involves exposing equipment to sterilizing gases in an enclosed heated chamber, allowing the gases to penetrate small spaces. Liquid sterilization involves submerging equipment in sterilizing liquids, which is less effective than gas but can be used for low contamination. The document then outlines a procedure to study the sterilizing activity of different agents on E. coli and B. subtilis bacteria over time to learn about sterilization effectiveness.

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    Exp 6

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    27 views12 pages
    This document discusses the use of chemical agents for sterilization when physical sterilization methods are not suitable. It explains that chemical sterilization can use liquid or gaseous agents to kill microorganisms. Gaseous sterilization involves exposing equipment to sterilizing gases in an enclosed heated chamber, allowing the gases to penetrate small spaces. Liquid sterilization involves submerging equipment in sterilizing liquids, which is less effective than gas but can be used for low contamination. The document then outlines a procedure to study the sterilizing activity of different agents on E. coli and B. subtilis bacteria over time to learn about sterilization effectiveness.

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    This document discusses the use of chemical agents for sterilization when physical sterilization methods are not suitable. It explains that chemical sterilization can use liquid or gaseous agents to kill microorganisms. Gaseous sterilization involves exposing equipment to sterilizing gases in an enclosed heated chamber, allowing the gases to penetrate small spaces. Liquid sterilization involves submerging equipment in sterilizing liquids, which is less effective than gas but can be used for low contamination. The document then outlines a procedure to study the sterilizing activity of different agents on E. coli and B. subtilis bacteria over time to learn about sterilization effectiveness.

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    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    27 views12 pages

    Exp 6

    Uploaded by

    Mehul Verma
    This document discusses the use of chemical agents for sterilization when physical sterilization methods are not suitable. It explains that chemical sterilization can use liquid or gaseous agents to kill microorganisms. Gaseous sterilization involves exposing equipment to sterilizing gases in an enclosed heated chamber, allowing the gases to penetrate small spaces. Liquid sterilization involves submerging equipment in sterilizing liquids, which is less effective than gas but can be used for low contamination. The document then outlines a procedure to study the sterilizing activity of different agents on E. coli and B. subtilis bacteria over time to learn about sterilization effectiveness.

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    Download as pptx, pdf, or txt
    of 12

    Demonstration of Sterlization

    – Chemical agents of control


    Dr. Saranya Punniyakotti
    Associate Professor
    Lovely Institute of Technology
    Theory/Principle:
    • A wide variety of chemical agents display

    antimicrobial activity to some degree.

    • Even if Physical methods of sterilization are more

    appropriate for effective sterilization, it is not

    always appropriate to use for heat-sensitive

    materials like plastics, fiber optics, and biological

    specimens.
    • Under such conditions, chemical either in liquid
    or gaseous state can be used for sterilization.
    • However, it is crucial to ensure that the materials
    undergoing sterilization are compatible with the
    chemical being used.
    • Besides, it is important to adopt safety rules in
    the workplace safety during the use of chemical
    agents.
    Gaseous sterilization
    • Gaseous sterilization involves the process of exposing equipment or devices to

    different gases in a closed heated or pressurized chamber.

    • Gaseous sterilization is a more effective technique as gases can pass through a tiny

    orifice and provide more effective results.

    • Besides, gases are commonly used along with heat treatment which also facilitates

    the functioning of the gases.

    • However, there is an issue of release of some toxic gases during the process which

    needs to be removed regularly from the system.

    • The mechanism of action is different for different types of gases.


    Liquid Sterilization
    • Liquid sterilization is the process of sterilization which
    involves the submerging of equipment in the liquid
    sterilant to kill all viable microorganisms and their
    spores.
    • Although liquid sterilization is not as effective as
    gaseous sterilization, it is appropriate in conditions
    where a low level of contamination is present.
    Aim: To study the activity of some sterilizing agents and to learn the
    importance of time, germicidal concentration, and microbial species
    in sterilization.
    Equipment and Apparatus Required: incubator.
    Materials Requirement: Nutrient agar plates Sterile, empty tubes Sterile 10-
    ml pipettes (cotton plugged) Sterile 1.0-ml pipettes (cotton plugged) Bulb or
    other aspiration device for pipette 5% sodium hypochlorite (bleach); 0.05%
    sodium hypochlorite Absolute alcohol, 70% alcohol 3% hydrogen peroxide, 24-
    hour nutrient broth culture of Escherichia coli, Three- to six-day-old broth culture
    of Bacillus subtilis.

    Learning objectives: To learn the chemical agents of control in


    sterilization.
    Outline of Procedure:
    1. Select one of the chemical agents provided. Pipette 5.0 ml of the
    solution into a sterile test tube.

    2. To the 5 ml of disinfectant, add 0.5 ml of the E. coli culture. Gently


    shake the tube to distribute the organisms uniformly. Note the time.
    3. Divide a nutrient agar plate into four sections with a marking pen
    or pencil. At intervals of 2, 5, 10, and 15 minutes, transfer one
    loopful of the disinfectant-culture mixture to a section of the
    nutrient agar plate. Label each plate with the name of the organism,
    the disinfectant, and its concentration (e.g., E. coli, 1% phenol).
    Label each section of the plate with the time of exposure (e.g., 2
    minutes, 5 minutes, etc.).
    4. Using the same concentration of the same disinfectant, repeat steps 1 to 3
    with the culture of B. subtilis.
    5. Inoculate one-half of a nutrient agar plate directly from the E. coli culture
    and the other half from the B. subtilis culture. Label each half with the name
    of the organism and the word Control.
    6. Incubate all tubes at 35°C for 48 hours.
    OBSERVATION
    Results:
    • Read all plates for growth () or no growth (). Record your
    results in the chart.

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