Praktikum Mikrobiologi Gram Negatif

8/9/2019 Praktikum Mikrobiologi Gram Negatif

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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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Praktikum Mikrobiologi Gram Negatif