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    Activity No. 7.

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    ALEXA JOIE CONCEPCION ARMONIA
    The document describes standard methods for obtaining pure cultures of microorganisms, including the streak plate method, pour plate method, and spread plate method. The streak plate method involves streaking a loopful of microbes across an agar plate to isolate individual colonies. The pour plate method uses serial dilutions and molten agar to isolate microbes from a mixed culture. The spread plate method distributes a fixed volume of inoculum evenly across an agar surface using an L-shaped rod. These techniques allow microorganisms to be separated from a mixed population and cultivated alone as pure cultures, which is necessary for the study of microbes in microbiology.

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    Activity No. 7.

    Uploaded by

    ALEXA JOIE CONCEPCION ARMONIA
    51 views3 pages
    The document describes standard methods for obtaining pure cultures of microorganisms, including the streak plate method, pour plate method, and spread plate method. The streak plate method involves streaking a loopful of microbes across an agar plate to isolate individual colonies. The pour plate method uses serial dilutions and molten agar to isolate microbes from a mixed culture. The spread plate method distributes a fixed volume of inoculum evenly across an agar surface using an L-shaped rod. These techniques allow microorganisms to be separated from a mixed population and cultivated alone as pure cultures, which is necessary for the study of microbes in microbiology.

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    The document describes standard methods for obtaining pure cultures of microorganisms, including the streak plate method, pour plate method, and spread plate method. The streak plate method involves streaking a loopful of microbes across an agar plate to isolate individual colonies. The pour plate method uses serial dilutions and molten agar to isolate microbes from a mixed culture. The spread plate method distributes a fixed volume of inoculum evenly across an agar surface using an L-shaped rod. These techniques allow microorganisms to be separated from a mixed population and cultivated alone as pure cultures, which is necessary for the study of microbes in microbiology.

    Copyright:

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

    Activity No. 7.

    Uploaded by

    ALEXA JOIE CONCEPCION ARMONIA
    The document describes standard methods for obtaining pure cultures of microorganisms, including the streak plate method, pour plate method, and spread plate method. The streak plate method involves streaking a loopful of microbes across an agar plate to isolate individual colonies. The pour plate method uses serial dilutions and molten agar to isolate microbes from a mixed culture. The spread plate method distributes a fixed volume of inoculum evenly across an agar surface using an L-shaped rod. These techniques allow microorganisms to be separated from a mixed population and cultivated alone as pure cultures, which is necessary for the study of microbes in microbiology.

    Copyright:

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

    WMSU-ISMP-GU-002.

    00
    Effective Date: 7-DEC-2016

    ISOLATION OF MICROORGANISMS
    Activity No. 7

    Microorganisms in nature are diverse and occur as a complex mixed population. A


    study of these microorganisms requires that they be separated from each other and
    cultivated in an artificial environment. This is called isolation. Different techniques are
    employed to isolate the different species from the mixture and cultivate them as pure
    cultures. A pure culture consists of a population of cells derived from a single cell.

    Streak plate is a very simple method of isolation if done properly. Plating method can
    also be used, and in addition, can approximate the variable organisms in the population.
    The use of enrichment or selective medium will enhance growth of the microorganisms or
    interest while inhibiting or killing the other types of microorganisms. The procedures
    intended in the section are the most common method of isolation.

    I. Objective/s:

    a. Demostrate the standard methods of obtaining a pure culture


    b. Appreciate the importance of pure cultures in the field of microbiology
    c. Distinguish the features of bacterial growth

    II. Materials:

    Each group will need the following:

    13 sterile petri dishes 1 wire loop


    3 sterile 9-ml water blanks 1 water bath
    in screw cap tubes Nutrient agar
    6 1-ml sterile pipets culture broth
    1 L-shaped sealed glass tubing

    III. Procedures:

    A. STREAK PLATE METHOD:

    a. Pour melted NA or CWA, following aseptic technique in a petri dish and allow the
    agar to solidify.
    b. Sterilize the wire loop and get a loopful of microorganisms of interest from the
    petri dish with mix culture done in the last activity.
    c. Streak very gently over the surface of the agar, taking care not to destroy it, as in
    figure below.

    38
    WMSU-ISMP-GU-002.00
    Effective Date: 7-DEC-2016

    d. Incubate the dish invertedly and examine the plate after 48 hours.
    e. Examine your previous petri dish further. Choose another colony and get a loopful
    f. This time streak this in an agar slant.
    g. Incubate and examine tube after 48 hours.

    B: POUR PLATE METHOD

    a. Liquefy medium in a water bath.


    b. Obtain three 9-ml water blanks in screw cap tubes and label each tube
    from 1 to 3.
    c. Secure a broth mix culture from your instructor.
    d. With a sterile pipet, transfer one ml from the stock culture to tube #1.
    Cover and shake vigorously.
    e. From tube #1, transfer 1 ml to tube #2. Mix.
    f. Again transfer 1 ml from tube #2 to tube #3. Shake and mix

    g. Using a pipet, transfer 1 ml from each tube to a duplicate sterile plates.


    Pour melted afar and mix as in the below.

    h. Allow the agar to solidify and incubate as in A4.


    i. Examine after 48 hours.

    C. SPREAD PLATE METHOD:

    a. Pour melted NA or CWA into each of six sterile petri dishes and allow to solidify.
    b. Transfer to duplicate plates 0.1 ml from each tube used in pour plate technique.
    c. Sterilize an L-shaped glass rod by dipping in alcohol and flaming until alcohol has
    burned off.
    d. Spread the inoculum evenly on the surface of the agar with an L-shaped rod.

    39
    IV. OBSERVATIONS:

    1. Write down your observations after 48 hours


    2. Describe the growth of microorganisms by referring to the cultural characteristics
    of bacterial growth incorporated in the previous activity, in terms of the following:

    1. For agar strokes:

    a. Amount of growth – scanty, moderate or abundant


    b. Form – filiform, echinulate, beaded, spreading or rhizoid

    2. For Agar colonies:

    a. Form – punctiform, circular, filamentous, rhizoid or irregular


    b. Elevation – effuse, flat, raised or convex
    c. Margin – entire, endulate, erose, filamentous or curled

    40

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