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    Kupang, 18 Februari 2016

    PENJELASAN PRAKTIKUM

    MIKROBIOLOGI BLOK KARDIOVASKULER

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    Optochinresistant

    Bile Soluble -

    Blood agar

    Tes Katalase

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    Mannitol SaltAgar

    DNAse Test

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    BacitrasinResistant

    Tes Katalase

    Blood Agar

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    vobiocin Resistant

    Mannitol Salt Agar

    atalase Test

    Koagulase Test

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    OptochinSensitive

    Bile Soluble + Blood agar

    Tes Katalase

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    BacitrasinSensitive

    Blood Agar

    Tes KatalaseCAM Test

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    V

    NovobiocinSensitive DNAse Test

    Blood agar Mannitol Salt

    Agar

    O!acilinResistant

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    STAPHYLOCOCCUS

    Frequent Human Infection Rare Human Infection

    S. aureus S. capitis

    S. epidermidis S. caprae

    S. hemolyticus S. saccharolyticus

    S. lugdunensis S. warneri

    S. saprophyticus S. pateuriS. hominis

    S. auricularis

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    STAPHYLOCOCCUS SP

    Bulat dengan diameter

    kira 1 µm

    Bersusun berkelompoksecara tidak beraturan

    Gram positif 

    Non-motil Non spora

    Katalase +

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    STAPHYLOCOCCUS

    AUREUS Bakteri gra !

    "eng#a$i%kan pigen %ip&'#r&

    S. aureus tera$uk bakteri &$&t&%eran

    Fakt&r )iru%en$i

    K&agu%a$e !

    Lek&$i*in

    +nter&t&k$in

     T&k$in Sin*r&a S&k T&k$ik-1

    He&%$in

    Su*a# banak ka$u$ ".SA

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

    around the colonies

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    CATALAS+  T+ST

    Differentiate Staphylococcifrom Streptococci andnterococci !catalasenegati"e#

    Detect presence of cytochromeo$idase % &'('  (' + &'(

    )est procedure%

    Slide catalase * place grothfrom colonies on glass slide,add . hydrogen pero$ide,"igorous bubbling seen !+"e#

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    Tube catalase – pick growthfrom media (not blood agar),immerse in 3% hydrogenperoxide

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    COA/ULAS+ T+ST

    )o differentiate S. aureus from other

    Staphylococcus sp

    S. aureus ha"e bound coagulase and /free0 

    coagulase

    Bound coagulase !/clumping factor0# present

    on surface of cell all

    /free0 coagulase - e$creted e$tracellularly

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    SL+ COA/ULAS+ T+ST

    Slide coagulase test

    Detect bound

    coagulase

    eact directly ith

    fibrinogen in plasma

    )est procedure% 2i$

    groth suspension ith

    D)3-rabbit plasma,obser"e agglutination

    !+"e#

    Slide coagulase test. Rapid method foridentifying S. aureus . Saline control onleft

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    COA/ULAS+ T+ST

    Some S. aureus do not produce bound

    coagulase, some coagulase-negati"e staph

    produce eak slide coagulase test !S.delphini, S. schleiferi #

    2ust confirm ith tube coagulase

    Do not take groth from mannitol salt agar *

    high salt content cause autoagglutination

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     TUB+ COA/ULAS+

    )ube coagulase

    4ndirect detection ofe$tracellular coagulase

    $creted coagulasedetect /coagulase-reacting factor0 in

    plasma

     form acomple$

    5omple$ ill react ithfibrinogen to form fibrin!/clot formation0#

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    MEDIUM MSA

    Beee3tra't 140 g

    Pept&ne 5i'& &r P&%pept&ne 5BBL 1040g

    7aC%940 g

    "annit&%1040 g

    P#en&% re*04029 g

    Agar

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    Mannitol salt agar - Mannitol fermentation indicated bychange in phenol red indicator to yellow

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    DNASE TEST

     T#e purp&$e i$ t& $ee i t#e i'r&be 'anu$e DNA  a$ a $&ur'e & 'arb&n an* energ&r gr&:t#4 U$e & DNA  i$ a''&p%i$#e* b

    an en;e 'a%%e* DNase4

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    NOVOBIOCIN SENSITIVITY

    U$eu% &r pre$upti

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    CHARACTERISTIC  S. aureus S. epidermidis S. saprophyticus

    Pigment Yellow towhite

    White White to palegray

    Hemolysis + ± -

    Coagulase prou!tion Yes "o "o

    #annitol$ermentation

    Yes "o "o

    "o%o&io!in sensiti%ity Sensiti%e Sensiti%e Resistan!e

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    STREPTOCOCCUSPHYLOGENETIC

    GROUPSPECIES LANCEFIELD GROUP TYPE OF

    HAEMOLYSISA

    Po!enic !rou" Str pyogenes

    Str. agalactiae

    Str. equisimilis

    A

    #

    C

    $

    $

    $

    Miti% !rou" Str. pneumoniae

    Str mitis

    Str. oralis

    Str. sanguis

    Str. gordonii

    O

    O

    Not i&entifie&

    H

    H

    '

    '

    '

    '

    '

    An!ino%u% !rou" Str. anginosusStr. intermedius

    G( F )an& A* ''

    Sa+i,ariu% !rou" Str. salivarius - None

    #o,i% !rou" Str. bovis D . or none

    Mutan% !rou" Str. mutans

    Str. sabrinus

    Not &e%i!nate&

    Not &e%i!nate&

    None

    None

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    /.A"-STA7

    Gram-positi"espherical or o"oid

    cocci Diplococci to long

    chains !in brothcultures#

    old cultures or deadbacteria mayappear gram-negati"e

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    STREPTOCOCCUSPYOGENES

    /ra !

    Berbentuk rantaiang pan=angna

    ber%ik,

    *engan teperatur&ptiu ?@C

    FAKTOR VIRULENSI

    He&%i$in

    Pr&tein ",F,*an /

    Kap$u% #ia%ur&nik

    Sub$tan$i C *anantigen ebran$it&p%a$a

    +k$&t&k$in

    Sprea*ing a't&r5pr&teina$e,$trept&kina$e,

    nuk%ea$e, *%%

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    H+"OLYSS

    6-hemolysis * complete hemolyis of rbc

    &emolysin induced

    2arker for pathogenicity

    7-hemolysis * greenish discolouration,partial hemolysis

    &ydrogen pero$ide induced * o$idi8eshaemoglobin to green methaemoglobin

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    H+"OLYS7

    &emolyse red blood cells

    Beta-hemolytic group 3 S. pyogenes *produce ' types of hemolysin

    Streptolysin S * obser"ed as complete lysisaround colonies on blood agar9 (ksigenstabil9 non-imunogenik

    Streptolysin (

    inacti"ated by o$ygen ! (ksigen-labil#

    4munogenik

     3ntibodies against streptolysin ( !3S)(#

    formed folloing infection

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    Alpha-hemolysis –greenish discoloration of blood agar

    CU!UR"C#ARAC!"R$S!$CS

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    %eta-hemolysis – clearing around colonies

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    BACT.AC7

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    BACT.AC7

    SUSC+PTBLTY :resumpti"e

    identification of Grp

     3 6-hemolyticstreptococci

    !susceptible to

    bacitracin#

    ; bacitracin

    disc * 8one of

    inhibition 1; mm or

    larger !susceptible#

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    CAMP TEST

    CHRISTIE ATKINS MUNCH-PETERSEN

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    CAMP TEST

    CA"P te$t

    CA"P a't&r

    pr&*u'e* ba=&rit & S.agalactiae

    A't$nergi$ti'a%%:it# beta#e&%$in

    pr&*u'e* b S.

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    +7TFCATO7 OF 7O7-B+TA-H+"OLYTC

    ST.+PTOCOCC   (ptochin test ?or identification of

    S. pneumoniae<

    (ptochin disks

    applied on

    streaked surface of

    blood agar 

    @ones A1= mm

    ith mm disc *

    sensiti"e

    !S. pneumoniae#

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    FF+.+7TATO7 OF 7O7-B+TA-H+"OLYTC ST.+PTOCOCC

    Streptococcu Optoc!"#ucept

    B"$eo$u%$e

    B"$e ecu$"#

    S. pneumoniae & & -

    S. bovis - - &

    Ot!er '"r"()# *roup - - -)

    a &ccasional strains may produce wea' reaction

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    BILE ESCULIN TEST

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    BILE SOLUBILITY TEST

    Prin'ip%e

    S. pneumoniae pr&*u'e a $e%-%$ing en;e t& in#ibit t#egr&:t#

     T#e pre$en'e & bi%e $a%t a''e%erate t#i$ pr&'e$$

    Pr&'e*ure

    A** ten part$ 510 % & t#e br&t# 'u%ture & t#e &rgani$ t& bete$te* t& &ne part 51 % & 2 7a *e&3'#&%ate 5bi%e int& t#e te$ttube

    7egati

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    BILE SOLUBILITY TEST

    .e$u%t$

    P&$iti

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    e.g. S. epidermidis

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    ba"teri aerob # a*a%a# bakteri angenggunakan &k$igen &%eku%ar $ebagaiterina% a$ept&r e%ektr&n $e%aa re$pira$i4

    ba"teri anaerob# a*a%a# bakteri angti*ak enggunakan &k$igen &%eku%ar$ebagai terina% a$ept&r e%ektr&n4 Bakteriini en*apat energi *engan e%akukanerenta$i atau *engan e%akukan

    re$pira$i anaer&bik4

    ba"teri $a"ultati$ anaerob# a*a%a#g&%&ngan bakteri ang bi$a en*apatenergi *engan re$pira$i atau erenta$i,

    tergantung pa*a kea*aan %ingkunganna4

    Ber*a$arkan Pr&*uk$i

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    Ber*a$arkan Pr&*uk$ien;i

    %& O"sidase ber*a$ar *i#a$i%kanna atau ti*ak&k$i*a$e, bakteri *ibagi ata$ bakteri positi$ o"sidase *an negati$ o"sidase4

    '& (e)olisin a*a%a# en;i ang *apat ee'a#kan

    eritr&$it ang ter*apat pa*a e*iu4 Ber*a$ar#e&%i$i$ $e%-$e% *ara# e*iu *i$ekitar k&%&nina,aka bakteri *i bagi ata$ 14 Ba"teri he)olisin *ang enebabkan erit&$it %i$i$ tetapi ebran $e%ti*ak ter%epa$, *an 24 Ba"teri he)olisin , *iana

    ter=a*i %i$i$ $e% eritr&$it ang $epurna4 & Koagulase  ang enebabkan penggupa%an

    p%a$a bi$a *i#a$i%kan atau ti*ak *i#a$i%kan &%e#bakteri4 Ber*a$arkan #a% ter$ebut bakteri *ibagi ata$14 Ba"teri positi$ "oagulase  *an 2 ba"teri

    negati$ "oagulase4

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    ANY +UESTION,,,,