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    Penjelasan Praktikum Mikrobiologi Blok Kardiovaskuler: Kupang, 18 Februari 2016

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    Eunike_ois
    This document provides instructions for a microbiology laboratory practical on cardiovascular block microorganisms. It describes tests to identify different bacterial species, including Staphylococcus, Streptococcus, and their characteristics. Identification is based on Gram stain morphology, catalase and coagulase production, hemolytic activity, growth on selective media like mannitol salt agar, and sensitivity or resistance to antibiotics. Common pathogens like S. aureus, S. epidermidis, and S. pyogenes are distinguished by their phenotypic properties. The document also provides descriptions of identification techniques such as catalase, coagulase, DNAse and optochin tests.

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    Penjelasan Praktikum Mikrobiologi Blok Kardiovaskuler: Kupang, 18 Februari 2016

    Uploaded by

    Eunike_ois
    167 views54 pages
    This document provides instructions for a microbiology laboratory practical on cardiovascular block microorganisms. It describes tests to identify different bacterial species, including Staphylococcus, Streptococcus, and their characteristics. Identification is based on Gram stain morphology, catalase and coagulase production, hemolytic activity, growth on selective media like mannitol salt agar, and sensitivity or resistance to antibiotics. Common pathogens like S. aureus, S. epidermidis, and S. pyogenes are distinguished by their phenotypic properties. The document also provides descriptions of identification techniques such as catalase, coagulase, DNAse and optochin tests.

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    PENJELASAN PRAKTIKUM

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    This document provides instructions for a microbiology laboratory practical on cardiovascular block microorganisms. It describes tests to identify different bacterial species, including Staphylococcus, Streptococcus, and their characteristics. Identification is based on Gram stain morphology, catalase and coagulase production, hemolytic activity, growth on selective media like mannitol salt agar, and sensitivity or resistance to antibiotics. Common pathogens like S. aureus, S. epidermidis, and S. pyogenes are distinguished by their phenotypic properties. The document also provides descriptions of identification techniques such as catalase, coagulase, DNAse and optochin tests.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    167 views54 pages

    Penjelasan Praktikum Mikrobiologi Blok Kardiovaskuler: Kupang, 18 Februari 2016

    Uploaded by

    Eunike_ois
    This document provides instructions for a microbiology laboratory practical on cardiovascular block microorganisms. It describes tests to identify different bacterial species, including Staphylococcus, Streptococcus, and their characteristics. Identification is based on Gram stain morphology, catalase and coagulase production, hemolytic activity, growth on selective media like mannitol salt agar, and sensitivity or resistance to antibiotics. Common pathogens like S. aureus, S. epidermidis, and S. pyogenes are distinguished by their phenotypic properties. The document also provides descriptions of identification techniques such as catalase, coagulase, DNAse and optochin tests.

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    PENJELASAN PRAKTIKUM

    MIKROBIOLOGI BLOK KARDIOVASKULER


    Kupang, 18 Februari 2016

    Optochin
    resistant

    Bile Soluble -

    Blood agar

    Tes Katalase

    Mannitol Salt
    Agar

    DNAse Test

    Bacitrasin
    Resistant

    Blood Agar

    Tes Katalase

    ovobiocin Resistant

    Mannitol Salt Agar

    Katalase Test
    Koagulase Test

    Bile
    Optochin
    Soluble + Sensitive

    Tes Katalase

    Blood agar

    Blood Agar

    Bacitrasin
    Sensitive

    Tes Katalase

    CAMP Test

    V
    Blood agar

    Mannitol Salt
    Agar

    Novobiocin
    Oxacilin
    Sensitive DNAse Test
    Resistant

    STAPHYLOCOCCUS
    Frequent Human Infection
    S. aureus
    S. epidermidis
    S. hemolyticus
    S. lugdunensis
    S. saprophyticus

    Rare Human Infection


    S. capitis
    S. caprae
    S. saccharolyticus
    S. warneri
    S. pateuri
    S. hominis
    S. auricularis

    STAPHYLOCOCCUS SP

    Bulat dengan diameter


    kira 1 m

    Bersusun berkelompok
    secara tidak beraturan

    Gram positif

    Non-motil

    Non spora

    Katalase +

    STAPHYLOCOCCUS
    AUREUS

    Bakteri gram +

    Menghasilkan pigmen lipochrom

    S. aureustermasuk bakteriosmotoleran

    Faktor Virulensi
    Koagulase +
    Lekosidin
    Enterotoksin
    Toksin Sindroma Syok Toksik-1
    Hemolysin

    Sudah banyak kasus MRSA

    S. aureus colonies on sheep blood agar. Zone of beta-hemolysis


    around the colonies

    CATALASE TEST

    Differentiate Staphylococci
    from Streptococci and
    Enterococci (catalase
    negative)

    Detect presence of cytochrome


    oxidase : H2O2 O2 + H2O

    Test procedure:

    Slide catalase place growth


    from colonies on glass slide;
    add 3% hydrogen peroxide;
    vigorous bubbling seen (+ve)

    Tube catalase pick growth


    from media (not blood agar),
    immerse in 3% hydrogen
    peroxide

    COAGULASE TEST

    To differentiate S. aureus from other


    Staphylococcus sp

    S. aureus have bound coagulase and free


    coagulase

    Bound coagulase (clumping factor) present


    on surface of cell wall

    free coagulase - excreted extracellularly

    SLIDE COAGULASE TEST

    Slide coagulase test

    Detect bound
    coagulase

    React directly with


    fibrinogen in plasma

    Test procedure: Mix


    growth suspension with
    EDTA-rabbit plasma;
    observe agglutination
    (+ve)

    Slide coagulase test. Rapid method for


    identifying S. aureus. Saline control on
    left

    COAGULASE TEST

    Some S. aureus do not produce bound


    coagulase; some coagulase-negative staph
    produce weak slide coagulase test (S.
    delphini, S. schleiferi)

    Must confirm with tube coagulase

    Do not take growth from mannitol salt agar


    high salt content cause autoagglutination

    TUBE COAGULASE

    Tube coagulase

    Indirect detection of
    extracellular coagulase

    Excreted coagulase
    detect coagulasereacting factor in
    plasma form a
    complex

    Complex will react with


    fibrinogen to form fibrin
    (clot formation)

    MEDIUM MSA

    Beef
    extract1.0 g

    Peptone (Difco) orPolypeptone(BBL)10.0


    g

    NaCl
    75.0 g

    Mannitol
    10.0 g

    Phenol red
    0.025 g

    Agar

    Mannitol salt agar - Mannitol fermentation indicated by


    change in phenol red indicator to yellow

    DNASE TEST

    The purpose is to see if the microbe can


    useDNAas a source of carbon and energy
    for growth. Use ofDNAis accomplished by
    an enzyme calledDNase.

    NOVOBIOCIN SENSITIVITY

    Useful for presumptively distinguishing


    Staphylococcus saprophyticusfrom
    other CoNS

    CHARACTERISTIC

    S. aureus

    Pigment

    Yellow to
    white

    White

    White to pale
    gray

    Coagulase production

    Yes

    No

    No

    Mannitol
    fermentation

    Yes

    No

    No

    Sensitive

    Sensitive

    Resistance

    Hemolysis

    Novobiocin sensitivity

    S. epidermidis S. saprophyticus

    STREPTOCOCCUS
    PHYLOGENETIC
    GROUP

    SPECIES

    LANCEFIELD GROUP

    TYPE OF
    HAEMOLYSISA

    Pyogenic group

    Str pyogenes
    Str. agalactiae
    Str. equisimilis

    A
    B
    C

    Mitis group

    Str. pneumoniae
    Str mitis
    Str. oralis
    Str. sanguis
    Str. gordonii

    O
    O
    Not identified
    H
    H

    Anginosus group

    Str. anginosus
    Str. intermedius

    G, F (and A)

    Salivarius group

    Str. salivarius

    None

    Bovis group

    Str. bovis

    or none

    Mutans group

    Str. mutans
    Str. sabrinus

    Not designated
    Not designated

    None
    None

    GRAM-STAIN

    Gram-positive
    spherical or ovoid
    cocci

    Diplococci to long
    chains (in broth
    cultures)

    old cultures or dead


    bacteria may
    appear gramnegative

    STREPTOCOCCUS
    PYOGENES

    Gram +

    Berbentuk rantai
    yang panjangnya
    bervariasi

    Ada yang berkapsul


    hialuronik

    Hemolisin

    Protein M,F,dan G

    Kapsul hialuronik

    Substansi C dan
    antigen membran
    sitoplasma

    Eksotoksin

    Spreading factor
    (proteinase,
    streptokinase,
    nuklease, dll)

    Bersifat fakultatif
    anaerob
    Tumbuh subur pada
    keadaan
    mikroaerofilik,
    dengan temperatur
    optimum 37C

    FAKTOR VIRULENSI

    HEMOLYSIS

    -hemolysis complete hemolyis of rbc

    Hemolysin induced

    Marker for pathogenicity

    -hemolysis greenish discolouration;


    partial hemolysis

    Hydrogen peroxide induced oxidizes


    haemoglobin to green methaemoglobin

    HEMOLYSIN

    Hemolyse red blood cells

    Beta-hemolytic group A S. pyogenes


    produce 2 types of hemolysin

    Streptolysin S observed as complete lysis


    around colonies on blood agar, Oksigen
    stabil, non-imunogenik

    Streptolysin O

    inactivated by oxygen ( Oksigen-labil)

    Imunogenik

    Antibodies against streptolysin O (ASTO)


    formed following infection

    CULTURE
    CHARACTERISTICS

    Alpha-hemolysis greenish discoloration of blood agar

    Beta-hemolysis clearing around colonies

    BACITRACIN
    SUSCEPTIBILITY

    Presumptive
    identification of Grp
    A -hemolytic
    streptococci
    (susceptible to
    bacitracin)

    0.04U bacitracin
    disc zone of
    inhibition 10 mm or
    larger (susceptible)

    CAMP TEST
    CHRISTIE ATKINS MUNCH-PETERSEN

    CAMP TEST

    CAMP test

    CAMP factor
    produced by
    majority of S.
    agalactiae

    Act
    synergistically
    with beta
    hemolysin
    produced by S.

    IDENTIFICATION OF NONBETA-HEMOLYTIC
    STREPTOCOCCI
    Optochin test

    For identification of
    S. pneumoniae.

    Optochin disks
    applied on
    streaked surface of
    blood agar

    Zones >14 mm
    with 6 mm disc
    sensitive
    (S. pneumoniae)

    DIFFERENTIATION OF NON-BETAHEMOLYTIC STREPTOCOCCI

    Streptococcus

    Optochin
    suscept

    Bile
    soluble

    Bile esculin

    S. pneumoniae

    S. bovis

    Other viridans group

    -a

    Occasional strains may produce weak reaction

    BILE ESCULIN TEST

    BILE SOLUBILITY TEST

    Principle:

    S. pneumoniae produce a self-lysing enzyme to inhibit the


    growth

    The presence of bile salt accelerate this process

    Procedure:

    Add ten parts (10 ml) of the broth culture of the organism to be
    tested to one part (1 ml) of 2% Na deoxycholate (bile) into the test
    tube

    Negative control is made by adding saline instead of bile to the


    culture

    Incubate at 37oC for 15 min

    Record the result after 15 min

    BILE SOLUBILITY TEST

    Results:

    Positive test appears


    as clearing in the
    presence of bile while
    negative test appears
    as turbid

    S. pneumoniae soluble
    in bile whereas S.
    viridans insoluble

    e.g. S. epidermidis

    bakteri aerob : adalah bakteri yang


    menggunakan oksigen molekular sebagai
    terminal aseptor elektron selama respirasi.

    bakteri anaerob: adalah bakteri yang


    tidak menggunakan oksigen molekular
    sebagai terminal aseptor elektron. Bakteri
    ini mendapat energi dengan melakukan
    fermentasi
    atau
    dengan
    melakukan
    respirasi anaerobik.

    bakteri fakultatif anaerob: adalah


    golongan bakteri yang bisa mendapat
    energi dengan respirasi atau fermentasi,
    tergantung pada keadaan lingkungannya.

    Berdasarkan Produksi
    enzim
    1. Oksidase: berdasar dihasilkannya atau tidak
    oksidase, bakteri dibagi atas bakteri positif oksidase
    dan negatif oksidase.

    2. Hemolisin adalah enzim yang dapat memecahkan


    eritrosit yang terdapat pada medium. Berdasar
    hemolisis sel-sel darah medium disekitar koloninya,
    maka bakteri di bagi atas: 1). Bakteri hemolisin ,
    yang menyebabkan eritosit lisis tetapi membran sel
    tidak terlepas, dan 2). Bakteri hemolisin , dimana
    terjadi lisis sel eritrosit yang sempurna.

    3. Koagulase yang menyebabkan penggumpalan


    plasma bisa dihasilkan atau tidak dihasilkan oleh
    bakteri. Berdasarkan hal tersebut bakteri dibagi atas:
    1). Bakteri positif koagulase dan 2) bakteri
    negatif koagulase.

    ANY QUESTION????

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