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PRACTICAL
Identifcation oGram-Negative
bacteriumBambang Isbandrio
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Sasaran Belaar!a"asis#a da$at menelas%an tentang&
strategi identif%asi ba%teri Gram-negati
enis-enis tes untu% memasti%an s$esiesba%teri Gram-negati $en'ebab ine%si
menginter$retasi%an "asil $emeri%saan
sensitivitas AB Gram-negati
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(iagnosisine%si
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4
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Catalase
Presence o catalase en)'me #"ic" brea%sdo#n $ero*ide radicals
Place a e# dro$s o "'drogen $ero*ide solution
on a loo$ul o 'our isolate on a glass slide and#atc" or t"e a$$earance o bubbles- t"e' ma'be ver' small
Bubbles + $ositive, no bubbles+negative
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*idase test
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!otilit' test
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Presum$tive ollo# u$
Gram Negative Rods
Oxidase negativeOxidase negative
Catalase positiveCatalase positiveMotile/ NonmotileMotile/ Nonmotile
Oxidase Positive
Catalase PositiveMotile
One o the mem!ers oentero!acteriaceae
"i!rio orPse#domonas etc
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Mem!ers oentero!acteriaceae$
oxidase Negative
%scherichia
&le!siella Citro!acter 'almonella 'higella Prote#s
etc
Nontero!acteriaceae
Oxidase positive $
"i!riosPse#domona
s
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'%(%C)*"% M%+*,
STOOLSSTOOLS
Sabouraud
Genta-chloram
mannitol
salt
Mac
Conkey
Hektoen SS
%NR*C-M%N).RO)-'
Rappaport Selenite
Campylosel
Yersinia
Cin
Clostridium
difficile
C-ROMOG%N*C M%+*,
Candida
! SM !
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Isolation . Culture& Colon'
Si)eSi)e
S"a$eS"a$e
ColorColor Surace eaturesSurace eatures
Smoot" - Roug"Smoot" - Roug"
Trans$arenc'Trans$arenc'
/emol'sis/emol'sis
SmellSmell
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MacConey 7M,C8 ,gar
Peptone 9:;< Polypeptone =:2< (actose9 9:=< .ile salts5 =:94 acid/acid pl#s gas0 -5' ,l/, > alaline/acid ,l/, E g > alaline/acid pl#s gas
,l/, E g0 -5' > alaline/acid pl#s gas0
-5'
,l/, E g0 -5' 7w8 > alaline/acid pl#s
gas0 -5' 7wea8
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,/, E g
Escherichia coli
Klebsiella pneumoniae
Klebsiella oxytoca Enterobacter aerogenes
Enterobacter cloacae
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&/,
Serratia marcescens90 5
Yersinia enterocolitica5
964< o strains
5Non3lactose0 s#crose ermenter
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&/, E g0 -5'
Citrobacter freundii
Proteus vulgaris9
9Non3lactose0 s#crose ermenter
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,l/,
Shigella
Providencia
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,l/, E g
Salmonella serotype Paratyphi,
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,l/, E g0 -5'
Salmonella 7most serotypes8 Proteus mirabilis
Edwardsiella tarda
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,l/, E g0 -5' 7w8
Salmonella serotype )yphi
Bioc"emical
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Bioc"emicaltests
Macconey$ (F
Gram$ Gram Negative
rodsCatalase$ Positive
Oxidase @ Negative
One o the mem!erso
)'*$ ,/,0 Gas E0 -5' Negative
'*M$ -5'0Neg0 *ndole$ Positive:0Motile
Motility@ 7P8 Motile*t co#ld !e
%'C-%R*C-*,0 C*)RO.,C)%R0OR%N)%RO.,C)%R
Gl#cose $ Positive 7,cidand Gas8
(actose $ Positive
'#crose $ Negative / Positive
Mannitol $ Positive
*ndole $ Positive
MR $ Positive
"P $ Positive
Citrate $ Negative
rease $ Negative
(,O $ E3E
Melonate$ 3
&CN @ H
'o it is %scherichia coli
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API S'stem
-aemophil#s in1#enIae
Pse#domonas aer#ginosa
%scherichia coli
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(isteria monocytogenes(isteria monocytogenes
'taphylococc#s xylos#s'taphylococc#s xylos#s
,ctinomyces pyogenes
API S'stem
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Pseudomonasand related organisms
"erobic #ram-ne#ati$e nonfermentati$e rodsPseudomonas aeruginosa: opportunistic infections of
multiple sites
Burkholderia cepacia: RT infection in cystic fibrosis
patients, UTI, opportunistic infections
Burkholderia pseudomallei: opportunistic pulmonary
infections or sepsis
Stenotrophomonas maltophilia: opportunistic infections
Acinetobacter baumannii:opportunistic infections of RT
Moraxella catarrhalis: opportunistic RT infections
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Pseudomonas
Structure and%hysiolo#y
Gram-negative rods.
Motile wit polar flagella.
!bligate aerobe.
!"idase-positive.
#o not ferment carboydrates.
Resistant to multiple drugs.
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P. aeruginosa
$orms round colonies wit a
fluorescent greenis color, sweetodor, and -emolysis.
%yocyanin- nonfluorescent
bluis pigment&
pyoverdin- fluorescentgreenis pigment&
pyorubin, andpyomelanin
'ome strains ave a prominent
capsule (alginate).
Identification of P. aeruginosais usually based on o"idase test
and its colonial morpology: -emolysis, te presence of
caracteristic pigmentsand sweet odor, and growt at *+ o.
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