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IMVIC

The document discusses biochemical tests used to identify members of the Enterobacteriaceae family of bacteria, particularly the IMViC tests. The IMViC tests include Indole, Methyl Red, Voges-Proskauer, and Citrate tests. Each test examines different metabolic processes to determine if bacteria are capable of producing certain compounds or utilizing specific substrates. The results of the IMViC tests can be used to differentiate bacterial species within the Enterobacteriaceae family.

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Fenny Aulia Sugiana
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149 views

IMVIC

The document discusses biochemical tests used to identify members of the Enterobacteriaceae family of bacteria, particularly the IMViC tests. The IMViC tests include Indole, Methyl Red, Voges-Proskauer, and Citrate tests. Each test examines different metabolic processes to determine if bacteria are capable of producing certain compounds or utilizing specific substrates. The results of the IMViC tests can be used to differentiate bacterial species within the Enterobacteriaceae family.

Uploaded by

Fenny Aulia Sugiana
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPT, PDF, TXT or read online on Scribd
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BIOCHEMICAL TESTING

IMVIC are a method to identify


family of Enterobacteriaceae,
usually E.Coli (Indicator food or
drink are not good)

Enterobacteriaceae
Large family of bacteria
Gram negative rods
Capable of fermenting various
sugars

Many found in the intestines of human or


other mammals
Commensals, opportunists or pathogens
Some found in the environment
Example species:
Escherichia coli
Klebsiella pneumoniae
Citrobacter freundii
Enterobacter aerogenes

Proteus mirabilis
Salmonella typhi
Shigella dysenteriae
Yersinia enterocolitica

Previous Experience with


Enterobacteriaceae: EMB Media

Large amounts of acid


from lactose fermentation
cause the dyes to
precipitate on the colony
surface, producing a
black center or a green
metallic sheen (E. coli)

Smaller amounts of acid


production result in pink
coloration of the growth
(E. aerogenes)

Nonfermenting enterics
do not produce acid so
their colonies remain
colorless or take on the
color of the media (P.
vulgaris)

Biochemical Testing
Triple Sugar Iron (TSI): fermentation of sugars, sulfur
reduction
IMViC:
Indole: Break down the amino acid Tryptophan
Methyl Red: Glucose oxidation
Voges-Proskauer: Production of neutral end products
Citrate: Citrate fermentation
Urease: Hydrolyzation of Urea
Phenylalanine Deaminase: converts the amino acid
phenylalanine to phenylpyruvic acid
Nitrate Reductase: Reduction of nitrate (NO3) to nitrite

IMViC TESTS

A series of four tests consisting of:

Indole: Break down the amino acid Tryptophan


Methyl Red: Glucose oxidation
Voges-Proskauer: Production of neutral end
products
Citrate: Citrate fermentation

Used to differentiate the


Enterobacteriaceae
We will look at each test individually

Indole Test
(SIM:
Sulfide,Indole,Motility)
Identifies bacteria capable of producing indole
Some bacteria are capable of converting
tryptophan (an amino acid) to indole and pyruvic
acid by using the enzyme tryptophanase
Pyruvic acid can be converted to energy or used
to synthesize other compounds required by the
cell

Tryptophan

Indole Ring

Pyruvic Acid

Ammoni

Procedure:

Obtain 4 SIM Deep tubes


Inoculate by the stab method with
the following organisms:
E.coli,
P.vulgaris,
E. aerogenes K. pneumoniae

Indole Test Results:


If indole is produced, upon addition of
Kovacs Reagent (10 drops), a cherryred band forms on the surface of the
media

Motility (if present) can be seen


as growth of the bacteria away
from the stab line
Sulfur in the media may be
reduced to hydrogen sulfide
(H2S); this appears as a
blackening within the media

Methyl Red Test

Used to determine the ability of a


bacteria to oxidize glucose and produce
stable acid end products
Methyl red is a pH indicator (red at pH
less than 4.4 and yellow at a pH greater
than 6)
The combination medium used for this
test is the MR-VP (methyl red/VogesProskauer) broth

Acid production: positive methyl red


End products of neutral pH : positive Voges-Proskauer

Procedure:

Obtain 3 MR-VP broth tubes


Inoculate (using a loop) with the
following organisms:
E.coli K.pneumoniae
E.aerogenes

Results:

From the 3 MR-VP broths that you


inoculated, transfer 2 mLs from each
and place into 3 separate clean tubes
(set aside these aliquots for the VP test)
To the remaining, original tubes that you
inoculated add 5 drops of methyl red
indicator
A red color indicates that glucose has
been oxidized

Methyl Red Test Results:


A red color indicates
that glucose has been
oxidized.

Methyl red positive


tube on the right

Methyl red negative


tube on the left

Voges-Proskauer Test

Used to determine the ability of


microbes to produce nonacidic or
neutral end products
Remember that the MR-VP broth is a
combined medium used for two tests
Methyl Red and Voges-Proskauer
You have already inoculated the 3 MRVP broth tubes from the previous
procedure (Methyl Red Test) with
E.coli
K.pneumoniae E.aerogenes

Voges-Proskauer
Procedure:

To the aliquots of each broth culture separated


during the methyl red test, add:
10 drops of Barritts Reagent A (KOH); shake
10 drops of Barritts Reagent B (-naftol) ;
shake
Reshake the culture every 3 to 4 min.
It can take as long as 15 min. for a color
change to occur

Voges-Proskauer Results:
The presence of a deep rose color
after 15 minutes is indicative of nonacidic / neutral metabolic end products
and a positive VP test result.

Voges-Proskauer
positive on the right

Voges-Proskauer
negative on the left

VP Principle
This test is used to identify microorganisms that carry
out a review of 2,3-butanediol fermentation. If the
bacteria ferment carbohydrate to 2,3-butanediol as the
main products, the materials will be occurred build up
on media. Add KOH 40% and alphanaphtol 5% in
ethanol can determine acetoin (asetilmetilkarbonil), a
primer compounds in synthesis of 2,3-butanediol. To
Add KOH, the acetoin indicated by the change in color
pink broth. The color change more clarify with add
alpha-naphthol solution. The color change is clear on
part of closer air, BECAUSE 2,3-butanediol is oxidised
partially
become acetoin to make clear result of
reaction

Citrate Utilization Test

Used to determine if an organism


is capable of fermenting citrate
and using that citrate as its sole
carbon source

The ability of an organism to utilize


citrate occurs via the enzyme
citrase

Procedure:

Obtain 3 Simmons Citrate agar


slants
Inoculate these slants using the stab
and streak method with the
following organisms:
E.coli K.pneumoniae
E.aerogenes

Citrate Test Results:

Simmons Citrate agar utilizes


sodium citrate as its sole
carbon source

Bromthymol blue is included as


a pH indicator; the medium
initially is green

Organisms capable of using


citrate as a carbon source turn
the media Prussian blue.

Summary of IMViC Reactions

Page 199

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