penjelasan praktikum
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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,,,,