Exercise 11

BIOCHEMICAL ACTIVITIES OF MICROORGANISM

NAME: Bliss Nove Faith Y. Polley, BSN - 1C

INSTRUCTION: For the table below, describe and interpret the expected result and give example bacteria per test results.

Introduction Principle USE Expected Results/ example bacteria

Hydrolysis of Many bacteria The test microorganisms It helps distinguish

Starch produce extracellular are cultivated on starch- between species of the
enzymes used to containing agar plates in genera Corynebacterium,
catalyze chemical the starch hydrolysis test. Clostridium, Bacillus,
reactions outside of If the bacteria can Bacteroides,
the cell. In this hydrolyze starch, they do Fusobacterium, and
manner, nutrient so in the medium, Enterococcus spp.
sources, such as especially around their
starch, that are too growth, whereas the rest
large to be absorbed of the plate contains non-
through the cell hydrolyzed starch. Positive: After the addition of the iodine solution, if there is a
membrane can be Because the medium clear area around in the line of growth, this shows that the
broken down into does not change color bacterium has hydrolyzed the starch.
smaller molecules when organisms Negative:  A color of the medium that is either blue, purple,
and transported into hydrolyze starch, an or black is observed.
the cell via diffusion. iodine solution is added to  
Starch agar is a the plate after incubation Positive Example: Bacillus subtilis
simple nutritive as an indication. While Negative Example:E. Coli
medium with starch non-hydrolyzed starch
added.  Beef extract takes on a dark blue color A microbe capable of making extracellular enzymes will create a
and pancreatic digest when exposed to iodine, clear area in the plates spreading out from beyond the growth,
of gelatin provide its hydrolyzed end and it will seem clear because there is no longer any starch
nitrogen, vitamins, products do not. present to react with the iodine (since it has been hydrolyzed by
carbon and amino   the exoenzyme produced by the bacteria). A bacteria that does
acids. Agar is the As a result, clear not make exoenzymes, on the other hand, will produce a dark
solidifying agent and transparent zones form agar in which the starch is still there and has reacted with the
starch is the around the colonies that iodine. We can see that everything is brown up until the bacterial
carbohydrate. hydrolyze starch, while growth, indicating that that organism did not hydrolyze the starch.
  the rest of the plate takes Bacillus subtilis is very much an illustration of a bacteria that has
on a dark blue color as the capacity to hydrolyze starch.
iodine creates the colored
complex with starch.
Carbohydrate Bacterial ability to Carbohydrate It is indicated for
Fermentation generate organic fermentation asserts that determining
compounds by an organism's action on a microorganism
metabolizing carbohydrate substrate fermentation reactions,
particular causes the medium to particularly enteric bacilli
carbohydrates and become acidic, which and Enterococcus.
related substances is may be identified using a  
a frequently used pH indicator dye. The They're primarily utilized to
approach for process through which differentiate and
microorganism bacteria make energy is presumptively identify
identification. known as carbohydrate gram-negative enteric
Different fermentation fermentation. During bacilli based on
media are used to glycolysis, most microbes carbohydrate fermentation
distinguish organisms convert glucose to patterns. Positive: Indicative of a successful carbohydrate
depending on their pyruvate; however, other fermentation response is the appearance of a yellow tint in
capacity to ferment species use alternative the medium after some time has passed.
carbohydrates in the routes. A fermentation Negative:  A negative sugar fermentation response can be
basal medium. medium is made up of a identified by the absence of yellow color appears in the
Purple Broth is used base medium and a product.
to investigate single carbohydrate for Positive Example:
carbohydrate fermentation (glucose, Escherichia coli
fermentation lactose, sucrose,
processes, mannitol, etc.). The Negative Example: Pseudomonas
particularly in the medium, on the other
identification of gram- hand, contains a variety The indicating dye, phenol red, turns red at pH 7 as an outcome
negative enteric of pH indicators. A of carbohydrate fermentation. When acid is produced during
bacteria that have Durham tube is placed in fermentation, the pH drops and the indicator dye turns yellow,
been given the each tube to capture gas indicating that the bacterium is responsible for converting the
carbohydrates of produced by metabolism, carbohydrate. Gas created during fermentation is contained in
interest. Vera was the in addition to a pH the Durham tube and appears as a bubbles in the upper third.
one who came up indicator to measure the When the bacteria run out of glucose, they turn to peptone,
with the purple formation of acid from which causes the pH to rise, lighting up of the indicator. All
media. The FDA fermentation. members of the Enterobacteriaceae family, which are glucose
recommends this Differentiating between fermenters, are examples of bacteria that can ferment
medium for sugar bacterial groups or carbohydrates.
fermentation species is aided by the
research. carbohydrate
fermentation patterns
displayed by different
organisms.
Hydrolysis of Gelatin is a collagen- The ability of an organism This test is used to
Gelatin derived protein to produce extracellular determine an organism's
generated from proteolytic enzymes ability to make
vertebrate connective (gelatinases) that liquefy gelatinases.
tissues. When gelatin, a component of  
collagen is heated in vertebrate connective This test can help you
water, it produces a tissue, is determined by distinguish between
substance called this test. Serratia, Proteus, Bacillus,
collagen. The Gelatinases hydrolyze Clostridium,
presence of gelatin into polypeptides Pseudomonas, and
gelatinases is in the first reaction, and Flavobacterium species.  
detected via gelatin polypeptides are then   Positive: At a temperature of 4 degrees Celsius, the
hydrolysis. Some transformed into amino This test distinguishes inoculation tube should be partially or totally liquid within 14
bacteria produce acids in the second pathogenic days, but the control tube should be completely solid. Clear
gelatinases, which reaction. The cell absorbs Staphylococcus aureus areas around gelatinase-positive colonies serve as an
are proteases that the amino acid and uses it from non-pathogenic indicator of gelatin hydrolysis on plates.
hydrolyze or digest for metabolic reasons. Staphylococcus Negative: At a temperature of 4 degrees Celsius, the tube
gelatin and are A nutritional gelatin epidermidis, which is completely solidified.
secreted medium is used to test gelatinase negative.  
extracellularly. the presence of  
Gelatinase production gelatinases. When an This test can be used to Positive Example: Staphylococcus aureus
is employed as a organism creates distinguish gelatin-positive
presumptive test for gelatinase, the enzyme Serratia and Proteus Negative Example: Pseudomonas aeruginosa
the identification of a hydrolyzes the gelatin in organisms from other
variety of organisms, the growth media to members of the Some microorganisms that have been able to break down gelatin
including liquefy it. Enterobacteriaceae family. remained liquid, while those that were unable to break down
Staphylococcus sp.,   gelatin repolarized and became solid. The negative outcome will
Enterobacteriaceae, Bacillus anthracis, B. produce no liquid, whereas the positive result will produce one.
and certain gram- anthracis, B. anthracis, B. This indicates that the bacteria can break down the gelatin.
positive bacilli. anthrac Gelatinase- Gelatinases, an enzyme produced by bacteria to break down
positive bacteria include gelatin, can be discovered via gelatin hydrolysis. In the first
cereus and numerous process, gelatin is hydrolyzed into polypeptides, which are
other species of the subsequently converted into amino acids in the second reaction.
genus, as well as The amino acid is absorbed by the cell and used for metabolic
Clostridium tetani and purposes. The amount of gelatinases is determined using a
Clostridium perfringens. nutritious gelatin medium. Gelatinase is produced by an
organism and hydrolyzes the gelatin in the growth media to
liquefy it. Bacillus anthracis and Bacillus cereus are two bacteria
that create gelatinases and produce a good outcome in gelatin
hydrolysis. Clostridium perfringens, Bacillus subtilis, and
Clostridium perfringensTetani.
The The ability of certain Tryptophan is an amino To differentiate Proteus
production of bacteria to acid that microorganisms mirabilis (indole negative)
Indole breakdown the amino that produce the from all
acid tryptophane into tryptophanase enzyme other Proteus species
indole, which can deaminate and (indole positive).
accumulates in the hydrolyze. Reductive  
medium, is deamination of tryptophan To
demonstrated in this produces indole, which is differentiate Klebssiellapn
test. The identification then converted to eumoniae (indole
of Enterobacteria indolepyruvic acid. The negative)
relies heavily on the deamination reaction, in from Klebsiellaoxytoca (in
indole synthesis which the amine (-NH2) dole positive). Positive: Within a few seconds of introducing the reagent,
assay. The majority group of the tryptophan   the presence of red color inside the iodine solution layer of
of E. coli strains M. molecule is eliminated, is To the agar thick was an indication that the reaction was
coli, E. coli, E. catalyzed by differentiate Citrobacterfre successful.
rettgeri, E. coli, E. tryptophanase. Indole, undii (indole negative) Negative:  Reagent layer will stay yellow or will be slightly
coli, E. coli, E. coli, E. pyruvic acid, ammonium from Citrobacterkoseri (ind hazy in appearance.
coli, E With the (NH4+), and energy are ole positive).
emission of indole, the reaction's end  Positive examples: Bacillus alvei, pathogenic E. coli, several
Morgani and products. As a coenzyme, Shigella strains, Enterococcus faecalis, and V. cholerae
Providencia species pyridoxal phosphate is
break down the required. Negative examples: Actinobacillus spp.,
amino acid   Aeromonassalmonicida, Alcaligenes sp., most Bacillus sp.,
tryptophan. This is The solution goes from Bordetella sp., Enterobacter sp., most Haemophilus sp.,
done through a yellow to cherry red when most Klebsiella sp., Neisseria sp., Mannheimiahaemolytica,
system of intracellular indole is coupled with Pasteurellaureae, Proteus mirabilis, P.
enzymes known as Kovac's Reagent (which
"tryptophanase," comprises hydrochloric Tryptophanase, an enzyme capable of metabolizing tryptophan
which is employed as acid and p- indole, is expressed by some bacteria. The addition of the
part of the IMViC dimethylaminobenzaldehy Kovacs reagent causes the indole product (but not tryptophan) to
procedures, which de in amyl alcohol). The react and turn a red color near the tube's top. An indole positive
are tests that are red colour will appear in result indicates that the bacteria produces the enzyme
used to identify an oily coating on top of tryptophanase. Tryptophan will be degraded into indole,
between members of the broth since amyl ammonia, and pyruvic acid as long as tryptophanase is available,
the alcohol is not water and Kovacs reagent can react with indole to give the red hue
Enterobacteriaceae soluble. near the tube's top.When there is no red color, but there is a
family. yellowish tint on top, this could indicate that the test is negative
When testing non- for indole, and so the enzyme tryptophanase is not created by
fermenters and those bacteria. Escherichia coli and Escherichia fergusonii are
anaerobes, a version two microorganisms that test positive in this test.
of this test called
Ehrlich's reagent
(which uses ethyl
alcohol instead of
isoamyl alcohol and
was invented by Paul
 Ehrlich) is employed.
Reduction of Other than ambient In a nitrate-containing Although all members of
Nitrate to oxygen, anaerobic broth, a heavy inoculum the Enterobacteriaceae
Nitrites metabolism needs an of the test organism is family reduce nitrate,
electron acceptor incubated. The nitrate some of them also
(O2). The final reductase enzyme is metabolize nitrite to
electron acceptor for produced by organisms produce other chemicals.
many gram-negative that convert nitrate in the It is thus utilized to
bacteria is nitrate. broth to nitrite, which can distinguish
The nitrate reduction then be further reduced to Enterobacteriaceae
test determines the nitric oxide, nitrous oxide, bacteria that produce the
amount of nitrate or nitrogen. nitrate reductase enzyme
reductase produced, The nitrate reduction test from Gram negative
which results in works by detecting nitrite bacteria that do not
nitrate reduction and its ability to generate generate the enzyme.
(NO3). a red compound when it  
Bacterial species are combines with sulfanilic In some species, nitrate
classified according acid to form a complex reduction may be linked to
to their ability to (nitrite-sulfanilic acid), anaerobic respiration.
convert nitrate to which then reacts with a -  
nitrite or nitrogenous naphthylamine to form a It is used in differentiating Positive: The nitrate reagents cause the medium to take on a
gases. red precipitate (prontosil), Mycobacterium red color after they are added.
which is a water-soluble   Negative: A putative indicator that the nitrate concentration
azo dye. Identifying Neisseria has been decreased below the nitrite level
However, red color is only species and distinguishing
formed when nitrate is them from Moraxella and
present in the medium. Kingella The nitrate
The absence of a red tint reduction test is an
in the medium after important test for
adding sulfanilic acid and distinguishing between N. Positive example:Staphylococcus carnosus
-naphthylamine indicates gonorrhoeae and N. Negative example: Enterobacteriaceae
that nitrite is not present. gonorrhoeae.
  gonorrhoeae and K. The nitrate reduction test determines whether a nitrate reductase
There is two explanations gonorrhoeaedenitrificans, enzyme is produced, resulted in nitrate reduction. The nitrate
for this observation. especially when K. reductase enzyme is produced by organisms that transform
1. The nitrate may denitrificans strains are nitrate in the broth to nitrite, which can then be further reduced to
not have been present. In stained nitric oxide, nitrous oxide, or nitrogen. When nitrite combines with
reduced; the smears, K. denitrificans sulfanilic acid to generate a complex (nitrite-sulfanilic acid), it
strain is nitrate- appears to be gram- reacts with a -naphthylamine to form a crimson precipitate
negative. negative diplococci. (prontosil), which would be a water-soluble azo dye. However,
2. The nitrate may   when nitrate is added to the medium, it produces a red tint.Upon
have been Facilitating species applying sulfanilic acid and -naphthylamine to the medium, there
reduced to nitrite identification of is no red color, indicating that nitrite is not present.
which has then Corynebacterium. Actinomycesisraelli, Actinomycesisraelli, Actinomycesisraelli,
been completely Actinomycesisraell A. naeslundii S. odontolyticusVeillonella
reduced to nitric species, epidermidis
 oxide, nitrous
oxide, or nitrogen
which will not
react with the
reagents that
react with nitrite;
the strain is
nitrate-positive.
 
When nitrite isn't
detected, it's time to see if
the organism has reduced
nitrate to something other
than nitrite. Indirectly, this
can be accomplished by
adding a small amount of
Zinc powder to the
culture. The reduction of
nitrate to nitrite is
catalyzed by zinc powder.
The development of a red
color in response to the
addition of Zinc indicates
that the organism was
unable to reduce nitrate,
implying that the test
organism is incapable of
doing so. If no color
change happens after the
addition of zinc, the
organism is a nitrate
reducer and has reduced
nitrate to one of the other
nitrogen molecules.
 
Note: Before inoculation,
a Durham tube is placed
in the nitrogen broth to
detect degradation of the
broth as demonstrated by
gas generation in the
tube, as well as to identify
denitrification by
organisms that produce
gas via other pathways.
Urease In 1946, Christensen Decarboxylation of amino
Activity created Urea Agar for acids results in urea.
the differentiation of Ammonia and CO2 are
enteric bacteria. The produced when urea is
urease test is used to hydrolyzed. The pH shift
detect an organism's is observed by the color
capacity to break change of phenol red
urea via the enzyme from light orange at pH
urease synthesis. 6.8 to magenta (pink) at
pH 8.1, as ammonia
production alkalinizes the
medium. Within 24 hours,
urease-positive microbes
color the entire medium
pink.
Negative organisms
generate no color change
or yellow as a result of
acid production, and
weakly positive organisms
can require several days.
Triple Sugar Most bacteria have The triple sugar-iron agar
Iron Agar the ability to ferment test, which uses Triple
carbohydrates, Sugar Iron Agar, is used
particularly sugars. to distinguish between
Among them, each species based on
bacteria can ferment carbohydrate
only some of the fermentation patterns and
sugars, while it hydrogen sulfide
cannot ferment the generation variances. The
others. Thus, the creation of gas and a shift
sugars, which a in the color of the pH
bacteria can ferment indicator from red to
and the sugars, which yellow indicate
it cannot is the carbohydrate
The ability to hydrolyze
urea with the enzyme
urease is utilized to
distinguish species in this
test.
 
This test can be used to
identify a variety of
Enterobacteriaceae
genera and species,
including Proteus,
Klebsiella, some Yersinia
and Citrobacter species, Positive: A favorable reaction is indicated by any extent of
and some pink in the reaction.
Corynebacterium species. Negative: The amount of germs present in the sample that
  was obtained is negligible.
It can also be used to
identify bacteria that Positive examples: Proteus, Klebsiella, Pseudomonas, and
produce the urease Staphylococcusspp
enzyme, such as Negative example: Escherichia coli
Cryptococcus spp.,
Brucella, Helicobacter The premise that urea medium, either broth or agar, includes
pylori, and many more. urea and also the pH indicator phenol red, can be used to
  analyze the results of urease activity. Many species can produce
To detect the presence of the urease enzyme, which catalyzes the breakdown of urea in
H. pylori, this test is the presence of moisture to produce two molecules of ammonia
conducted directly on and carbon dioxide. Ammonium carbonate is formed when
gastric biopsy samples . ammonia combines with carbon dioxide and water, making the
medium alkaline and discoloring of the indicator form orange-
yellow to vivid pink. Cryptococcus, Brucella, and Helicobacter
pylori are among the bacteria that manufacture the urease
enzyme.
The test is mostly used to
distinguish
Enterobacteriaceae
bacteria from other gram-
negative rods.
It can also be used to
distinguish between
Enterobacteriaceae based
on their sugar
fermentation processes.
A- An alkaline/acidic reaction, also known as a red
characteristic of the fermentation. slant/yellow butt response, is evidence that solely dextrose
bacteria and thus an fermentation has occurred.
important criterion for B- It shows the ferment of dextrose, lactose, and/or sucrose
its identification. when there is an acid/acid reaction (also known as a yellow
The Triple Sugar slant/yellow butt).
Iron (TSI) test is C- An alkaline/alkaline reaction, sometimes known as a "red
a microbiological test slant" or "red butt" reaction: Because there are no
named for its ability to carbohydrates present, fermentation cannot take place. 
test a D- Triple sucrose iron agar. A, Acid slant or acid butt with air
microorganism’s but no H2S present (A/A). B, Alkali slant/acid butt, negative
ability to ferment for gas and positive for H2S (K/A H2S+). C, Alkali slight
sugars and to angle but, no gases, no Hydrogen sulfide (K/K). D, a tube
produce hydrogen with no inoculum.
sulfide. An agar slant  
of a special medium Positive example: Salmonella Typhimurium Bacteria
with multiple sugars
constituting a pH- Negative example: Enteric bacilli
sensitive dye (phenol
red), 1% lactose, This can occur at the same time and can be used to interpret the
1% sucrose, outcome of this activity. When one of carbohydrates is
0.1% glucose, as well fermented, the pH drops, changing the color of the medium from
as sodium reddish orange to yellow. A dark crimson color indicates
thiosulfate and ferrou alkalinization of the peptones. Some bacteria convert sodium
s sulfate or ferrous thiosulfate in the medium to hydrogen sulfide (H 2S), which
ammonium sulfate is reacts with ferric ions to generate iron sulfide, a black, intractable
used for carrying out deposit. Salmonella and Shigella are two bacterium examples.
the test. All of these
ingredients when
mixed together and
allowed solidification
at an angle result in a
agar test tube at a
slanted angle. The
slanted shape of this
medium provides an
array of surfaces that
are either exposed to
oxygen-containing air
in varying degrees
(an aerobic environm
ent) or not exposed to
air
(an anaerobic environ
ment) under which
fermentation patterns
of organisms are
 determined.
Citrate This test reveals Bacteria that grow on
Utilization whether bacteria can citrate agar produce
use citrate as their citrate-permease, an
only source of carbon enzyme that converts
for energy. The citrate to pyruvate.
presence of a citrate Pyruvate can then be
permease, which used to produce energy in
enables citrate the organism's metabolic
transport into the cycle. Growth indicates
bacteria, is required that citrate, an
for this ability. intermediate metabolite in
Citrate agar is used the Krebs cycle, is being
to determine whether used.
or not an organism The ammonium
can utilise citrate as a compounds are converted
source of energy. down to ammonia when
Citrate is the only bacteria metabolize
carbon source, and citrate, increasing
inorganic ammonium alkalinity. Above pH 7.6,
salts (NH4H2PO4) the bromthymol blue
are the only nitrogen indicator in the medium
supply in the medium. changes color from green
to blue due to the pH
change.
MR-VP Test In 1898, Voges and Some bacteria can use
Proskauer discovered glucose as a source of
that adding energy and convert it to
potassium hydroxide lactic acid, acetic acid, or
to colonies grown on formic acid as a
certain media caused byproduct.
them to turn red.  
Harden later These bacteria convert
demonstrated that the glucose to pyruvic acid,
red color was caused which is then metabolized
by the synthesis of via the "mixed acid
acetyl-methyl pathway" to create the
This test is among a suite
of IMViC Tests (Indole,
Methyl-Red, Vogues-
Proskauer, and Citrate)
that are used to
differentiate among the
Gram-Negative bacilli in
the family
Enterobacteriaceae.
Positive: Along the slant, growth that is accompanied by a
change in color from green to bright blue.
Negative: There is neither growth nor a change in color; the
slant continues to be green.
 
Positive examples: Salmonella, Edwardsiella, Citrobacter,
Klebsiella, Enterobacter, Serratia, Providencia
Negative example: Escherichia coli
This test examines whether bacteria can use citrate as their sole
carbon source. Klebsiellapneumoniae and Proteus mirabilis are
two examples of bacteria that use citrate. Bacteria that develop
in citrate agar create citrate-permease, an enzyme that
transforms citrate to pyruvate. In the organism's metabolic cycle,
pyruvate is being used to generate energy. Growth is fueled by
citrate, a Krebs cycle intermediate molecule. The ammonium
molecules in citrate are converted to ammonia by bacteria, which
raises alkalinity. Above pH 7.6, the bromothymol blue indicator in
the medium turns blue instead of green.
This test is used to
determine which
fermentation pathway is
used to utilize glucose.
The paired MR-VP tests
were originally intended to
discriminate between
members of the
Enterobacteriaceae family,
but they are currently
employed to characterize Positive: Turns the color into red
carbinol. Barrit stable acid. The type of
improved the acid produced varies from
sensitivity of the test species to species and is
by adding alpha- determined by the
naphthol to the bacteria's particular
solution before enzymatic pathways. The
adding potassium resulting acid lowers the
hydroxide in 1936. pH to 4.5 or below, as
The methyl red (MR) shown by a shift in the
test identifies the color of methyl red from
creation of adequate yellow to red.
acid during glucose  
fermentation and the To see if an organism
preservation of creates
circumstances that acetylmethylcarbinol from
keep the pH of an old glucose fermentation, the
culture below 4.5, as Voges-Proskauer (VP)
evidenced by a test is utilized. In the
change in the color of presence of - naphthol,
the methyl red strong alkali (40 percent
indicator applied at KOH), and ambient
the conclusion of the oxygen,
incubation period. acetylmethylcarbinol is
By aliquoting sections transformed to diacetyl.
of the inoculated The - naphthol was not
medium to different included in the original
tubes, Clark and Lubs process, but Barritt
developed MR-VP discovered that it acts as
Broth, which allowed a color intensifier and
both the MR and VP must be introduced first.
tests to be done from The diacetyl and
the same inoculated quanidine-containing
media. chemicals in the broth's
peptones then condense
into a pinkish red
polymer.
2 pyruvate = acetoin +
2CO2
acetoin + NADH + H+ =
2,3-butanediol + NAD+
The test bacterium is
cultivated on a broth
medium containing
other bacteria groups, Negative: Turns the color into yellow
including Actinobacteria.  
Positive example: Escherichia coli
A culture will usually only Negative example:Enterobacter cloacae
be positive for one
pathway: either MR+ or
VP+. Escherichia coli is
MR+ and VP-. In
contrast, Enterobacteraero
genes and Klebsiellapneu
moniae are MR- and
VP+. Pseudomonas
aeruginosa is a glucose
nonfermenter and is thus
MR- and VP-.
Positive: In the event that the test tube becomes red, it
indicates that mixed acid fermentation was present
Negative: The color of the culture medium has not changed 
 
Positive example: Escherichia coli
Negative example: Pseudomonas aeruginosa
As shown by a change in colour of the methyl red indicator
applied at the end of the incubation time, the methyl red (MR)
test detects correct acid formation throughout glucose
fermentation and even the conservation of factors that keep the
pH of an old culture below 4.5. For the Methyl Red test, the pH
indicator methyl red is put into an inoculation tube of MR-VP
broth. The acids will overpower the buffers in the medium and
generate an acidic environment if the organism uses the mixed
acid fermentation pathway and produces stable acidic end-
products. If acidic end products are present when methyl red is
injected, the methyl red would remained red.
 glucose in the methyl red
test (MR test). When the
methyl red is given to the
broth culture and the
bacteria can use glucose
to produce a stable acid,
the color of the methyl red
changes from yellow to
red.
 
The mixed acid pathway
gives 4 mol of acidic
products (mainly lactic
and acetic acid), 1 mol of
neutral fermentation
product (ethanol), 1 mol
of CO2, and 1 mol of H2
per mol of glucose
fermented. The large
quantity of acids
produced causes a
significant decrease in the
pH of the culture medium.

References: 

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