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    Staphylococci: Exercise No. 9

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    dennise reyes
    This document provides instructions for identifying Staphylococcus bacteria species through a series of tests. It describes performing gram stains on S. aureus and S. epidermidis under light, high power, and oil immersion objectives. Tests are also described to observe hemolysis on blood agar plates, presence of catalase using hydrogen peroxide, coagulation of human plasma in coagulase testing, and mannitol fermentation using mannitol salt agar. The results of these tests can be used to differentiate S. aureus from S. epidermidis and other Staphylococcus species.

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    Staphylococci: Exercise No. 9

    Uploaded by

    dennise reyes
    20 views5 pages
    This document provides instructions for identifying Staphylococcus bacteria species through a series of tests. It describes performing gram stains on S. aureus and S. epidermidis under light, high power, and oil immersion objectives. Tests are also described to observe hemolysis on blood agar plates, presence of catalase using hydrogen peroxide, coagulation of human plasma in coagulase testing, and mannitol fermentation using mannitol salt agar. The results of these tests can be used to differentiate S. aureus from S. epidermidis and other Staphylococcus species.

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    WEEK 8

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    This document provides instructions for identifying Staphylococcus bacteria species through a series of tests. It describes performing gram stains on S. aureus and S. epidermidis under light, high power, and oil immersion objectives. Tests are also described to observe hemolysis on blood agar plates, presence of catalase using hydrogen peroxide, coagulation of human plasma in coagulase testing, and mannitol fermentation using mannitol salt agar. The results of these tests can be used to differentiate S. aureus from S. epidermidis and other Staphylococcus species.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    20 views5 pages

    Staphylococci: Exercise No. 9

    Uploaded by

    dennise reyes
    This document provides instructions for identifying Staphylococcus bacteria species through a series of tests. It describes performing gram stains on S. aureus and S. epidermidis under light, high power, and oil immersion objectives. Tests are also described to observe hemolysis on blood agar plates, presence of catalase using hydrogen peroxide, coagulation of human plasma in coagulase testing, and mannitol fermentation using mannitol salt agar. The results of these tests can be used to differentiate S. aureus from S. epidermidis and other Staphylococcus species.

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    of 5

    Name: Date: __________________

    Section: Score: __________________

    Exercise No. 9
    Staphylococci
    Staphylococci are normally found on the skin and mucous membranes, a well as in our
    environment. It is, therefore, necessary to determine whether given staphylococci isolate is
    pathogenic or not. There are no absolute criteria, which distinguish pathogenic form non-
    pathogenic strains, but pathogenic strains generally hemolyze red blood cells, produce catalase
    and plasma coagulase and ferment mannitol.
    Objectives:
    1. The student must be able to know and perform the methods and techniques of isolation
    2. and identification of bacteria.
    3. The student must be able to cultivate Staphylococcus sp.
    4. The students must be able to perform Gram staining and biochemical tests for
    Staphylococcus.
    5. The student must be able to differentiate the species of Staphylococcus based on their
    colonies and reactions to different biochemical tests.
    Materials:
    Bacterial suspension of S. aureus & S. epidermidis
    Blood Agar plate
    Glass slide
    Wire loop
    Alcohol lamp
    Compound microscope
    Sterile applicator stick
    Sterile test tubes
    Mannitol Salt Agar
    Human blood plasma
    Cedarwood oil
    Reagents:
    3% H2O2
    Gram stain reagents
    Procedures:
    A. Gram stain:
    1. Prepare a bacterial smear from the bacterial suspension. (always practice aseptic
    technique)
    2. Stain with Gram stain.
    3. Examine under LPO, HPO and OIO.
    S. aureus under LPO S. aureus under HPO S. aureus under OIO

    S. epidermidis under LPO S. epidermidis under HPO S. epidermidis under OIO


    B. Hemolysis Test:
    1. Divide with a glass market a blood agar plate into 2 sectors and streak on each sector
    one of the two organisms above. Label. Incubate at 37oC for 18 to 24 hours.
    2. Following incubation, read results. Hemolysis is indicated by a clear zone around the
    colonies on BAP.
    3. Study the hemolytic activity of the 2 organisms.
    4. Make necessary drawings.
    S. aureus Hemolysis Test S. epidermidis Hemolysis Test

    C. Catalase Test
    1. Emulsify a colony using sterile applicator stick in 1 drop of 3% H2O2 in a glass slide
    2. Observe for the production of gas bubbles
    3. Interpretation: Gas bubbles = positive; No gas bubbles = negative
    4. Make necessary drawing.
    S. aureus catalase test S. epidermidis catalase test

    D. Coagulase Test
    1. Place 0.5ml of fresh human plasma in a small sterile tube. Inoculate the tube with one
    loopful of Staphylococcus aureus. Do the same with the Staphylococcus epidermidis.
    2. Incubate at 37°C for 4 hours.
    3. Observe for coagulum formation.
    4. If no coagulum is formed, incubate further up to 18 hours.
    5. Interpretation: Clot formation = positive; No clot formation = negative
    S. aureus coagulase test S. epidermidis coagulase test

    A. Mannitol Fermentation Test


    1. Divide with a glass marker a MSA into 2 sectors and streak organisms on each sector.
    Label. Incubate.
    2. After incubation, read results. A change of color of the corresponding compartment of
    MSA from pink to yellow indicates fermentation of mannitol.
    3. Examine which specie ferments mannitol and make necessary drawing.
    Mannitol Fermentation test
    Experiment worksheet on Staphylococci
    I. Complete the table below.
    a. Indicate the diseases that is caused by the different specie of Staphylococcus and its treatment
    Staphylococcus specie Disease and Treatment
    characteristics
    S. aureus

    S. epidermidis

    S. saphrophyticus

    Reference:
    a. Indicate the use of the differential test for Staphylococcal specie
    Differential test Use
    Coagulase test

    Mannitol Fermentation test

    Tellurite Glycine test

    Polymyxin Sensitivity test

    Voges Proskauer test

    Reference:

    b. Identify the differential test applicable for the different Staphylococcal and the corresponding
    result
    Microorganism Differential test Result
    S. aureus vs. S.
    saphrophyticus

    S. aureus vs. S.
    epidermidis

    S. saphrophyticus vs. S.
    epidermidis

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