BIOCHEMI

CAL TEST
PURPOSE
ORGANISM
IDENTIFIED
REAGENT
POSITIVE
RESULT
NEGATIVE
RESULT
+
representative
- UNDERLYING PRINCIPLE
NOTES

representative
1 determines if organism 0.04U susceptible: resistant: no zone S. pyogenes, S. agalactiae, ** BACITRACIN inhibits synthesis of bacterial cell This method is helpful in
BACITRACIN is susceptible or bacitracin disk zone of inhibition of inhibition M. luteus S. aureus wall screening for group A
susceptibility S. pyogenes
test resistant to 0.04U of (Taxo A) < 10 mm streptococci in throat
bacitracin ** Group A streptococci are susceptible to low cultures
levels (0 .04 units) of bacitracin, whereas other
groups of β-hemolytic streptococci are resistant.
2 ID of Group D org. capable of bile esculin BROWN- no blackening of group D Viridans ** ESCULIN is a coumarin derivative of glycoside mainstay in identification
BILE streptococci growth in agar BLACK PPT agar slant enterococcus Streptococci; that contains glucose moiety and a glycone schemes for the
ESCULIN (Enterococcus & presence of 4% (esculin, 4% (blackening of and non- moiety. nonhemolytic, catalase-
TEST/Agar Non-enterococci) from NOTE: Growth enterococcus, E. coli negative, gram-positive
(BEA)
bile and can bile, ferric agar slant)
other Streptococci. citrate) alone does not ** Mahon*: Esculetin diffuses into the agar and cocci
hydrolize
constitute a growth: combines with ferric citrate in the medium to
esculin to
Mahon*: differentiate positive result. Enterococcus produce a black complex.
ESCULETIN
group D streptococci feacalis ** Growth indicates tolerance to 4% bile.
and enterococci from ** BEA (+) organisms - hydrolize exculin to
GROUP D
other gram-positive esculetin, which acts with ferric citrate to form
STREPTOCOCCI
cocci. brown-black ppt.
3 differentiates S. bile LYSIS OR intact colonies bile soluble: bile insoluble: ** BILE - capable of rapidly AMIDASE
Bile pnuemoniae from S. (sodium DISINTEGRATION S. E. faecalis dissolving/destroying colonies of S.
Solubility alpha-hemolytic pneumoniae desoxycholate) OF COLONIES pneumoniae pneumoniae.
test streptococci (Viridans) ** BILE SALT lower the surface tension bet. cell
and medium.
** Lysis depends on the presence of AMIDASE
(intracellular autolytic enzyme).

** Takes advantage of the very active S.

pneumoniae autocatalytic enzyme amidase.
** Under the influence of a bile salt or detergent,
the organism’s cell wall lyses during cell division.

** Mahon*: A suspension of S. pneumoniae in a

solution of sodium deoxycholate lyses, and the
solution becomes clear.
** Other α-hemolytic organisms do not undergo
lysis, and the solution remains cloudy.
4 differentiate sheep blood enhanced no enhanced enhanced beta-hemolysis ** S. agalactiae produces CAMP factor which “CAMP” - acronym from
CAMP S. agalactiae from
S. agalactiae
agar plate hemolysis by arrowhead arrowhead without acts synergistically with beta-lysin of S. aureus, the first letters of the
test other Beta hemolytic arrowhead zone formation of hemolysis: arrowhead: causing enhanced lysis RBCs. surnames of the
(Christie- Streptococci S. aureus of beta hemolysis colonies S. agalactiae S. pyogenes individuals who first
Atkins isolate at the juncture of ** IOW, enhances the lysis of sheep RBCs by described the reaction:
& 2 org staphylococcal BETA-LYSIN Christie, Atkins, and
Munch- Munch-Petersen.
Peterson) ARROWHEAD- ** CAMP factor - thermostable, antigenic,
SHAPED area of diffusible extracellular hemolytic protein. *CAMP inhibition
enhanced CAMP factor enlarges the zone of lysis formed by reaction (reverse CAMP
hemolysis where Staphylococcal beta hemolysis positive) = Inhibition of
the two streaks hemolysis by S. aureus
(staphylococcal where the two streaks
and approach each other. This
streptococcal) reaction is characteristic of
approach each Arcanobacterium
other. haemolyticum.
5 differentiate catalase catalase (+) 30% COPIOUS no or few bubbles Staphylococcus Streptococcus ** Aerobic organisms: produce H2O2 & O2- and *Note: add 18-hour old
Catalase (+) micrococcal & Staphylococcus Hydrogen BUBBLES & Micrococcus have catalase. colony to H2O2
test staphylococcal species and peroxide
from catalase (-) Micrococcus EFFERVESCENCE ** CATALASE - converts/ mediates the *do not use blood agar,
Streptococcal species breakdown of H2O2 into water & oxygen. The use TS agar, since blood
GAS conversion reaction produces gas in forms of has catalase and will give
PRODUCTION bubbles. a false (+) result.

*Add reagent to the

organisms, do not add
organism to reagent since
this may lead to false (+)
rxn.
6 differentiate S. aureus EDTA-rabbit CLOT of any No clot S. aureus other ** S. aureus produces 2 forms of coagulase. ** Negative slide test
Coagulase and prediction of plasma size; Staphylococci; must be confirmed with the
test
S. aureus
Staphylococcal ** 2 types of coagulase: tube test.
virulence macroscopic S. epidermis
clumping or clot (1) BOUND COAGULASE - "clumping factor"; ** False (+) result can be
formation; bound to bacterial cell wall and reacts directly to caused by citrate-utilization
fibrinogen resulting to precipitation of fibrinogen (+) organisms such as
Delost: white leading to cell clumping when bacterial Serratia, Pseudomonas,
fibrin clots in suspension is mixed with plasma. Enterococcus; these
plasma detected by SLIDE COAGULASE TEST organisms can use citrate
and release calcium, so
(2) FREE COAGULASE - extracellular protein clot occurs in the absence
enzyme that causes clot formation; detected using of coagulase.
TUBE COAGULASE TEST.
** Tubes should be
** Clotting mechanism: activation of CRF, checked for clot formation
forming coagulase-CRF complex. This complex intervals (usually @ 4 hrs.
reacts with fibrinogen forming the fibrin clot. & 24 hrs.) due to possible
production of fibrinolysin
MOST IMPORTANT DIFFERENTIAL TEST FOR (staphylokinase), which
ID OF S. aureus & PREDICTION OF may lyse the fibrin clot.
STAPHYLOCOCCAL VIRULENCE
There is now latex
agglutination test for
Staphylococcus aureus
which detects for clumping
factor and protein A
7 determines the ability of Serratia sp., DNase agar MEDIUM IS medium remains S. aureus; E. coli ** PALE-GREEN MEDIUM due to DNA-methyl
DNA organism to hydrolyze (toluidine blue COLORLESS green Serratia sp.; green complex.
Hydrolysis DNA Moraxella or methyl AROUND THE Moraxella
(DNase green as TEST catarrhalis; ** If organism growing in the medium hydrolyzes
catarrhalis,
Test Agar) differentiate organisms indicator ORGANISM Plesiomonas DNA, GREEN COLOR FADES and colony is
based on the shigelloides, surrounded by a COLORLESS ZONE.
production of S. aureus Aeromonas
deoxyribonuclease hydrophila; ** When DNA hydrolyzed, methyl green is
Stenotrophomona released and combines with highly polymerized
s maltophila, DNA at a 7.5 pH turning the medium colorless
S. pyogenes around the test organism.

** NO DEGRADATION OF DNA, medium

REMAINS GREEN.

8 presumptive Enterobacteria medium is BLACKENED no/slight E. faecalis E. coli ** Organisms is able to hydrolyze the glycoside
Esculin identification and ceae nonselective MEDIUM; blackening; esculin.
Hydrolysis differentiation of agar unlike BE
Enterobacteriaceae, hydrolysis
 media which is show loss of no loss of ** ESCULIN is hydrolyzed into esculetin which
determine if organism is selective due fluorescence fluorescence under reacts with Fe3+ and forms a DARK
able to HYDROLYZE to bile under wood's wood's lamp BROWN/BLACK PPT
the glycoside ESCULIN lamp
9 presumptive test for ID Staphylococcus nutrient gelatin PARTIAL OR complete Proteus Enterobacter ** organisms able to produce gelatinase; Determination of results
Gelatin of various organisms spp., medium TOTAL solidification of vulgaris; aerogenes; must be completed after
Hydrolysis (solidifying LIQUEFACTION tube at 4 deg C ** GELATIN - component of vertebrate connective refrigeration because
determines if Enterobacteriac agent is of inoculated tube B. subtilis E. coli tissue (liquid above 20degC). gelatin is liquid above 20
organism is eae, replaced with at 4 deg C degC
PROTEOLYTIC agar); within 14 days ** GELATINASE - extracellular proteolytic
some gram (+) (loss of gelling) enzyme that liquefy gelatin
determines the ability to bacilli gelatin deep,
produce proteolytic incubated at
enzymes (gelatinase) 35 degC
that liquefy gelatin (25degC) for
up to 14 days
10 determines the ability of to detect DEEP PURPLE colorless or slightly S. agalactiae; S. pyogenes ** end product of hydrolysis of hippuric acid by MEDIUM MUST CONTAIN
Hippurate organism to produce S. glycine - COLOR yellow pink color HIPPURICASE include GLYCINE and BENZOIC ONLY HIPPURATE,
Hydrolysis hippuricase which AGALACTIAE NINHYDRIN; Enterococcus ACID because ninhydrin might
splits hippuric acid into react with free amino acids
glycine and benzoic to detect ** GLYCINE is deaminated by oxidizing agent present in media.
acid; benzoic acid NINHYDRIN, which is reduced in the process.
- FERRIC An isolate that is
presumptive ID of CHLORIDE Mahon*: Subsequent addition of ninhydrin results hippurate positive and
variety of in the release of ammonia from the oxidative bile esculin negative has
microorganisms Ninhydrin: deamination of the α amino group in glycine as a very high probability of
3.5 g well as the reduced form of ninhydrin, being S. agalactiae.
Mahon*: To differentiate HYDRINDANTIN.
Streptococcus The ammonia reacts with residual ninhydrin and
agalactiae from other hydrindantin to produce a PURPLE-COLORED
βhemolytic streptococci COMPLEX. End product of ninhydrin oxidation
react to form a purple-colored product.
12 presumptive LAP disk RED COLOR No color change or E.faecalis Aerococcus ** Leucine-beta-naphthylamide-impregnated LAP disk is a rapid test
Leucine identification of CATALASE (-) (impregnated WITHIN 1 development of a viridans; disks – substrate for the detection of LEUCINE for the detection of the
Aminopep catalase-negative GRAM (+) with leucine MINUTE after slight yellow color AMINOPEPTIDASE. enzyme leucine
tidase gram-positive cocci COCCI Beta- Leuconostoc aminopeptidase.
adding
(LAP) Test napthylmide). ** After hydrolysis of the substrate by the enzyme,
cinnamaldehyde
cinnamaldehy reagent the resulting BETA-NAPHTHYLAMINE produces
de reagent. a red color upon addition of CINNAMALDEHYDE
REAGENT.
13 determine an Appearance of Appearance of Fermentation: ** FERMENTATION OF LACTOSE is
Litmus organism’s ability to Indicator Milk. Clostridium demonstrated when the litmus turns pink as a
Milk metabolize litmus milk (Litmus Dye). perfringens (gas result of acid production.
Medium (1) Acid or production);
differentiates (1) Acid (A) - alkaline (clot) - ** If sufficient acid is produced, CASEIN in the
microorganisms based PINK, MAUVE COAGULATION; Acid: milk is coagulated, solidifying the milk. With some
on various metabolic Lactobacillus organisms, the curd shrinks and whey is formed at
reactions in litmus milk, (2) (2) Acid acidophilus (clot the surface.
including fermentation, Alkaline(K) - (digestion) - formation);
reduction, clot BLUE Dissolution of clot ** Some bacteria hydrolyze casein, causing the
formation, digestion, with clear, Peptonization: milk to become STRAW COLORED AND
and the formation of (3) no change grayish, watery Pseudomonas RESEMBLE TURBID SERUM.
gas. - PURPLE fluid and a aeruginosa
(identical to shrunken, (clearing ** Some organisms reduce litmus, in which case
Litmus milk is also used uninoculated insoluble PINK Appearance of the medium becomes COLORLESS in the bottom
to grow lactic acid control); CLOT; indicator (litmus of the tube.
bacteria. dye))
(3) Alkaline
(Peptonization) -
 (4) White Dissolution of clot
(Decolorized) - with grayish,
Independent watery fluid and a
of pH change; clear, shrunken,
result of insoluble BLUE
reduction of CLOT
indicator.

14 differentiate gram- gram-positive, 6% DARK BLUE; No color change Micrococcus; Staphylococcus ** differentiation is based through the detection of Staphylococci should
Microdase positive, catalase- catalase-positive Tetratmethyl blue to purple - the enzyme OXIDASE. yield a negative color
Test positive cocci cocci paraphynylene blue color Micrococcus Staphylococcus change, except for S.
(Modified (micrococci from diamine luteus aureus ** In the presence of atmospheric oxygen, the sciuri, S. lentus, and S.
Oxidase) staphylococci). (Micrococcus) dihydro- oxidase enzyme reacts with the oxidase vitulus.
chloride with reagent and cytochrome C to form the colored
rapid method to dimethyl compound, INDOPHENOL.
differentiate sulfoxide
Staphylococcus from
Micrococcus spp.
15 determine whether an Some MRS Broth; GAS No growth Lactobacillus E. coli ** MRS broth contains sources of carbon,
organism forms gas Lactobacillus incubated for PRODUCTION nitrogen, and vitamins to support the growth of
Lactoba during glucose spp. and 24-48 hrs. @ (bubbles in lactobacilli and other organisms.
cillus fermentation. Leuconostoc 35degC Durham tube).
MRS spp. produce ** Selective medium - uses sodium acetate and
Broth Some Lactobacillus gas. Leuconostoc ammonium citrate to prevent overgrowth by
spp. and Leuconostoc contaminating organisms.
spp.— growth;
spp. produce gas.
gas production
** Growth is considered a positive result.
indicated by a
** A Durham tube may be added to differentiate
bubble in the
Lactobacillus spp. from Leuconostoc spp.
Durham tube.

Lactobacillus
spp.—growth;
no gas
production.
16 determine the effect of pneumococci; 6ug or 10ug zone of No zone of Streptococcus Streptococcus ** OPTOCHIN - lyses pneumococci (positive
Optochin optochin of Taxo P inhibition at inhibition pneumoniae pyogenes test), but alpha-streptococci are resistant
(P disk) (ethylhydrocupreine S. least 14 mm (negative test).
Susceptibil hydrochloride) on an (chemical
pneumoniae in diameter, with
ity Test organism; name: ethyl ** OPTOCHIN is an antibiotic that interferes with
6-mm disk
hydrocupreine the ATPase and production of adenosine
determine if organism is hydrochloride) triphosphate (ATP) in microorganisms.
susceptible or resistant
to optochin; ** The antibiotic inhibits the growth of a
susceptible organism, creating a clearing, or
to differentiate S. zone of inhibition, around the disk.
pneumoniae from
Viridians Streptococci *A zone of 14 to 16 mm is considered
susceptible and presumptive identification for
Streptococcus pneumoniae.
17 differentiate Enterobacteriaci Semisolid m. Positive: Acid Negative: RED or Fermenter: ** Nonfermentative bacteria are routinely tested Note: If the organism does
Oxidation microorganisms based aea (O-F medium) production (A) - alkaline (K) color in Escherichia for their ability to produce acid from six not grow at all in the OF
and on the ability to oxidize Glucose YELLOW. the deep. coli; carbohydrates (glucose, xylose, mannitol, lactose, medium, mark the rxn as
Fermenta or ferment specific mineral oil - sucrose, and maltose). no growth (NG).
tion carbohydrates. barrier to Weak-positive No color change Oxidizer
oxygen (Aw): color in both tube – Pseudomonas ** OF glucose is used to determine whether an Glucose fermenter – acid
of determine whether an organism ferments or oxidizes glucose. production in both tubes
remains about NON-OXIDIZER, aeruginosa
Medium organism uses b. purple NON- (fermentation can occur
(CDC the same as it
carbohydrate (purple to was before and FERMENTER ** no reaction occurs in either the TSI or OF with or without oxygen)
Method)
substrates to produce yellow) the inoculated Red or alkaline (K) glucose - non–glucose utilizer organism. **B&S
acid byproducts medium in control color in the deep Acid production in
Andrade’s tube becomes a ** Production of acid in the overlaid tube results anaerobic tube
Differentiate members acid fuchsin deeper red. in a color change and is an indication of
of family (pale yellow to FERMENTATION. Glucose oxidizer – acid in
Enterobacteriaceae, pink No change (NC) open, aerobic tube
(glucose fermenters) or neutral (N): ** Acid production in the open tube and color
from aerobic Phenol red There is growth change is the result of OXIDATION.
pseudomonads & (red – yellow) but no color
similar gram (-) bacteria b. blue (green change in 2
(nonfermenters) – yellow) tubes.
18 to determine if PYR disk and BRIGHT RED No color change or Enterococcus Streptococcus ** enzyme L-pyrrolidonyl arylamidase
l-Pyrrolido organism is capable of Streptococcus N,N- COLOR an orange color faecalis; S. agalactiae hydrolyzes the L-pyrrolidonyl-naphthylamide
– nyl producing the enzyme pyogenes & methlyaminoci WITHIN 5 pyogenes substrate to produce A-NAPHTHYLAMIN.
Arylamida L- namaldehyde
se (PYR)
MINUTES
pyrroglutamylaminopept ** Beta - naphthylamine can be detected in the
test
E. faecalis
idase (PYRase) presence of N,N-methylamino-cinnamaldehyde
reagent by the production of a bright red
presumptive precipitate
identification of group
A streptococci
(Streptococcus
pyogenes) and
enterococci by the
presence of the enzyme
L-pyrrolidonyl
arylamidase
19 determine the ability of pyruvate broth Indicator changes No color change; Enterococcus Streptococcus ** Pyruvate broth is a carbohydrate-free, nutrient-
Pyruvate an organism to utilize Enterococcus from green to yellow-green faecalis bovis; limited medium.
Broth pyruvate faecalis YELLOW indicates a weak
reaction and Enterococcus ** PYRUVIC ACID - added to the broth to
aids in the should be regarded faecium determine whether the microorganism is able
differentiation between as negative to use pyruvate, results to formation of metabolic
Enterococcus faecalis acids.
(positive) and
Enterococcus faecium ** Indicator: BROMOTHYMOL BLUE changes
(negative). from blue to yellow in presence of acid(decrease
in pH)
20 determine the ability of heart infusion VISIBLE No turbidity and no Enterococcus. Non Heart infusion broth & small amount of glucose
Salt an organism to grow in Enterococci broth with TURBIDITY in color change Enterococcus enterococcus. and bromocresol purple as the indicator for acid
Tolerance high concentrations 6.5% NaCl the broth, with or faecalis (growth) - Streptococcus production.
Test of salt. and without a color color change to bovis
bromocresol change from yellow.
Differentiate purple PURPLE TO (inhibition)- little
Enterococci (positive) YELLOW to no growth; no
from nonenterococci color change
(negative).
21 used to determine the P. aerugenosa Acetamide BLUE COLOR No color change P. aerugenosa Stenotrophomo ** Bacteria that can grow on this medium
Acetamide ability of an organism to medium nas maltophila deaminate acetamide to release ammonia.
utilization use acetamide as the
sole source of carbon. Incubated ** The production of ammonia results in a pH-
35oc for 7
driven color change of the medium (green→blue)
days
22 used to determine if an Acetate BLUE COLOR No growth or E. coli Shigella flexneri ** Breakdown of sodium acetate causes the pH of
Acetate organism can use medium/slant Medium becomes growth with no the medium to shift toward the alkaline range,
Utilization acetate as the sole alkaline (blue) indicator change to turning the indicator from green to blue.
Test source of carbon Incubated because of the blue
35oc for 7 growth of the
days organism

23 Used to determine the Malonate BULE COLOR ** Bacteria that can grow in the malonate broth
Malonate ability of an organism to broth can also use ammonia sulfate as nitrogen source.
utilization use sodium malonate
test as the sole source of (+) utilization of ammonium sulfate, causing a shift
carbon on the alkaline pH

24 a rapid test for the Moraxella Butyrate disks BLUE-COLOR No color change Moraxella N. gonorhoeae ** If the bromo-chloro-indolylbutyrate aka CATARRHALIS TEST
Butyrate detection of enzyme (Branhamella) (bromo-chloro- (Branhamella) impregnated disks is hydrolyzed by the enzyme, a
Disk butyrate esterase in catarrhalis indoyl during 5-minute catarrhalis blue-colored indigo compound is formed.
identifying Moraxella butyrate) incubation period
(Branhamella)
catarrhalis
25 used to determine the P. Certimide agar GROWTH P. aeruginosa E. coli ** CETRIMIDE – a toxic substance that inhibits
Cetrimide ability of an organism to slant the growth of many bacteria by causing the
aeruginosa
test grow in the presence of Icubated @ release of nitrogen and phosphorous, which slows
cetrimide 35oC for 7 or kills the organism.
days
for identifying P. ** P. aeruginosa is resistant to cetrimide
aeruginosa

primarily used to isolate

and purify
Pseudomonas
aeruginosa from
contaminated
specimens.
26 is used to determine the Good Growth No growth P. aeruginosa P. fluorescens
Growth at ability of an organism to
42°C grow at 42°C

27 Determine if an Microscopic: Darting motility Nonmotile: remain ** Can be determined by microscopic examination Some bacteria are motile
Motility organism is motile Hanging Drop – Campylobacter at the site of or by observing growth in a semi solid medium only RT
Testing Method & sp. inoculation.
flagellar stain 2 motility tubes may be
** An organism must possess flagella to be motile.
Tumbling end- inoculated. (1) RT; (2)
Semisolid
medium: SIM over-end 35oC
or MIO
medium (</=
0.4% agar

28 Determine the presence Pseudomonads
Oxidase of bacterial
Test CYTOCHROME
OXIDASE using the
oxidation of the
substrate tetramethyl-p-
phenylenediamine
dihydrochloride to
indophenol, a dark
purple-colored end
product.
Can be used to
differentiate
enterobacteriaciae (-)
from Pseudomonads
(+)
29 The X and V factor test Haemophilus
X and V is used to differentiate
factor Haemophilus spp.
Members of the genus
Haemophilus require
accessory growth
factors in vitro.
Some Haemophilus
spp. require X factor
(hemin) alone, V factor
(nicotinamide adenine
dinucleotide [NAD])
alone, or a combination
of the two.
conc – allow motility – Listeria In true motility, the organisms change in position
free spread of monocytogenes with respect to each other, often darting across
microorganism the field. (+ result)
s) Gliding motility
– In Brownian movement, the organisms may
Capnocytophaga appear quite active, but they remain the same
sp. relative position to other organisms or debris in
the field (- result)
Swarming
motility –
proteus sp.
0.5% - 1% Development of a Absence of color N. gonorrhoeae E. coli KOVAC’S METHOD Remove colony with
tetramethyl-p- DARK PURPLE platinum or wooden
phenylenedia COLOR within 10 Moraxella Ways to performs applicator
mine – 30 seconds - rub colony on a paper strip and add the reagent
seconds Most DO NOT USE NICHROME
dihyrochloride, - rub colony on a piece of paper containing the
pseudomonads LOOP! (contains iron→
prepared fresh reagent
- put a drop of reagent on the colony false (+)
daily Aeromonas
Do not use NA or MHA
Campylobacter because can lead to
inconsistent results
Pasteurella
** Members of the Genus Haemophilus require
spp. necessary growth factors in vitro.
** Some Haemophilus spp require X factor
(hemin) alone, V factor (NAD) alone, or a
combination of both.
** A lawn of the test organism is streaked onto
heart infusion agar, tryptic soy agar, Haemophilus
agar, or nutrient agar.
** The impregnated disks or strips (X, V, or XV)
are placed directly on the confluent inoculation,
allowing diffusion of the accessory growth factor
into the medium.
** The organisms will grow only around the disk
that provides the appropriate factor for growth of
the organism.
30 30% H202 Gas production, Neisseria
Bubbles or
Super - effervescence
oxol test
31 Determines if organism For differentiation 1.24ug of Zone of inhibition
Sulfameth is susceptible or of Streptococcal SXT Organism Bacitracin SXT
oxazole- resistant 1.24ug of groups Group A S R
trimethopr sulfamethoxazole- Group B R R
im trimethoprim (SXT) Not Group A or B; possibly C, F, or G R S
susceptibi
lity test
32 Neufeld- Can be used to identify S. S. Swelling capsules Streptococcus Viridans ** “quellung” swelling
quellung S. pnuemoniae which pneumoniae- of organism pneumoniae Streptococci
pneumoniae
test possesses capsule specific ** Makes the capsule swell of become prominent
(capsular- capsular
swelling antigen
test)
33 Determines the ability Tryptophan- RED OR No ring E. coli Klebsiella ** TRYPTOPHANASE – splits tryptophan to form ERLICH’S INDOLE TEST
Indole of organism to produce containing indole & deaminate tryptophan to inedole. Pyruvic uses extraction of indole
PINK RING
TRYPTOPHANASE media (SIM, acid, and ammonia from broth using xylene;
test tryptone or MORE PREFERRED since
peptone broth) ** Organisms that are indole (+) and nitrite more sensitive
reduction (+) is also CHOLERA-RED POSITIVE
Kovac’s KOVAC’S INDOLE TEST
reagent does not use extraction

Erlich
reagent (p-
dimethylamino
benzaldehyde)
34 Determine the ability of MRVP broth RED No color change E. coli Klebsiella Those organisms that carry out mixed acid MRVP broth – used to
Methyl organism to produce fermentation produce vast amounts of acid that determine by which
and maintain stable end Peptone will convert the methyl red indicator to red color bacteria metabolizes
red test products from glucose glucose broth glucose
fermentation
Methyl red The tests detect the end
(MIXED ACID (indicator) fermentation of glucose
PATHWAY) fermentation, in
35 Determine the ability of MRVP broth RED No color change Klebsiella E. coli Measures the production of acetoin. accordance with the
Vogues organism to produce pathway an organism uses
proskauer neutral red products 40% KOH or The addition of 40% potassium hydroxide followed to metabolize glucose.
tes (acetoin or cetyl methyl NaOH by a-naphthol results in red complex (neutral pH)
carbinol) from glucose which indicates a positive test Methyl red has reciprocal
fermentation Alpha- relationship with vogues-
naphthol a-naphthol – added as catalyst or color intensifier proskauer test
2,3 butanediol & (indicator) 40% KOH or NaOH – oxidized to diacetyl
acetoin
(BUTYLENE GLYCOL Diacetyl – forms red complexat relatively neutral
PATHWAY) pH
36 Determines the ability Simmons GREEN → Remains green Klebsiella E. coli Determines whether an organism can utilize False (+) may occur with
Citrate of organism to use citrate agar BLUE pneumoniea sodium citrate as sole source of carbon. an inoculum that is too
utilization SODIUM CITRATE as (bromothymol Ammonium salts are nitrogen source in the heavy.
test the sole source of blue) medium and utilization of these results in the
carbon, and inorganic release of ammonia, causing pH change.
ammonium salts as its Cristensen The bromothymol blue indicator turns the medium
only nitrogen source citrate medium from green to blue.
(phenol red)
 Incubated @ YELLOW → Growth in the Simmons citrate agar and change in
35oC for 7 PINK color of medium from green into blue
days
Growth in the Christensen citrate medium and
change in color of medium from yellow to pink

37 Determine the ability of MUG disk ELECTRIC Lack of E. coli P. aeruginosa ** BETA-D-GLUCORONIDASE – hydrolyzes
MUG test organism (E. coli) to (imoregnated BLUE fluorescence beta-d-glucorosid-uronic derivaties into aglycons
produce enzyme with 4- FLUORES- and D-glucoronic acid
BETA-D- methylbelli- CENCE
GLUCORONIDASE feryl-beta-D- ** The substrate 4-methylumbelliferyl-
glucoronide) -d-glucuronide is impregnated into the disk and
used to presumptively is hydrolyzed by the enzyme to yield the 4-
identify various genera methylumbelliferyl moiety, which fluoresces blue
of Enterobacteriaceae under long wavelength ultraviolet light.
and verotoxin-
producing Escherichia ** verotoxin-producing strains of E. coli do not
coli produce MUG, and a negative test result may
indicate the presence of a clinically important
strain.
38 Determine the ability of Sulfanilic RED Development of ** NITRATE REDUCTASE – enzyme that can If red color is not detected,
Nitrate organism to produce acid & RED COLOR after reduce nitrate into nitrite all available nitrate may
reduction NITRATE Alpha- addition of zinc have been reduced t nitrite
test REDUCTASE naphtylamine confirm a true ** Potassium → nitrite + sulfanilic acid → nitrite & and then completely
negative result alpha-naphtylamine → DIAZO RED DYE converted into nitrogen gas
ZINC (N2), nitric oxide (NO), or
** If no color develops, add small amount of zinc nitrous oxide (N2O). no
dust to distinguish whether there is true negative nitrite remains to react with
result, or if it was due only to further reduction of sulfanilic acid.
nitrite to nitrogen gas
All members of the
** Metallic zinc reduces nitrate to nitrite Enterobacteriaceae family
A red color after addition of zinc indicates that reduce nitrate, but some
nitrate was still in the broth (negative) members further
metabolize nitrite to other
** Absence of red color after addition of zinc compounds.
indicates that no nitrate was left (positive)
39 Biochemical BLACK Erysiphelothrix ** A bacterium utilizes the sodium thiosulfate
Hydrogen medium rhusiopathiae sulfur source to form H2S, a colorless gas.
sulfide (TSIA, LIA)
(H2S) Salmonella ** H2s combines with the indicator, FERROUS
productio Requires an Proteus SULFATE
n Test org. source of Citrobacter
sulfur and a Edwardsiella ** Numerous media demonstrate the production of
source of H2S (SIM media, MIO agar, HEA, SSA, TSIA,
metal KIA, LIA)
40 B- Determines if the O-nitrophenyl- YELLOW colorless Rapid/slow Non-lactose ** Organisms that are late or slow lactose
galactoside organism is a slow or beta-D- lactose fermenters fermenters may appear as non-lactose fermenters
& late lactose fermenter galactopyraoni fermenters on primary media
orthonitrop side
henyl-B-D- Determines the ability **Late lactose fermenter – lacks permease
galacto-
pyranoside
of organism to produce ** Lactose nonfermenter – lack the b-
(ONPG) the enzyme BETA- galactosidase
test GALACTOSIDASE
** PERMEASE – allows lactose to enter the cell
 ** B-GALACTOSIDASE – splits lactose; acts on
ONPG to form a yellow compound which indicates
that the organism is a lactose fermenter

** BETA GALACTOSIDASE converts ONPG into

orthonitrophenol
41 Determine if organism Urea LIGHT ORANGE No color change RAPID UREASE SLOW ** UREASE – hydrolyzes urea, causing of
Urea is able to produce agar/slant → MAGENTA Remains buff to (4 HOURS) UREASE (24 ammonia and CO2, causing shift of pH to alkaline
hydrolysis urease (Christensen (bright pink in yellow Proteus HRS) range
(urease) urea agar) Mahon) Providencia Klebsiella
test with phenol Morganella Enterobacter ** Ammonia reacts in soln to form ammonium
red as pH Helicobacter Yersinia carbonate, which increase the pH
indicator Citrobacter
Serratia ** This is detected by PHENOL RED in the
medium and turns the medium a bright pink color
Positive ghap
ine, wa la space ** CHRISTENSEN’S UREA AGAR is preferred
test medium
42 Determine if organism Phenylalanine GREEN Proteus E. coli ** Deamination of phenylalanine results in Modification: use of
Phenylala is capable of slant (0.2% Providencia production of PHENYLPYRUVIC ACID. tryptophan agar to detect
COLOR
nine deaminating phynylalanine) Morganella for tryptophan deaminase
deaminase phenylalanine into & 10% ferric ** DECARBOXYLASE – enzyme that removes
test phenyl pyruvic acid chloride the carbonyl (COOH) group from amino acids
(PAD)
** DEAMINASE – an enzyme that removes (NH2)
group from amino acids
43 Determines ability of Decarboxy- PURPLE yellow Lysine— Lysine— ** Such organism can decarboxylate or hydrolyze Testing 3 amino acids
Decarboxy organism to use amino lase broths
COLOR Klebsiella Citrobacter and amino acids to form amine, causing alkaline require 4 tubes act as
lase tests acids as energy and (Lysine, freundii— pH change control
change pneumoniae
(Moeller’s carbon sources ornithine, yellow
—yellow to
method) arginine) Lysine (d) →cadaverine
purple
Ornithine— Ornithine (d) → putrescine
Bromcresol Proteus Arginine (d) → putrescine
Ornithine—
purple & vulgaris— Arginine (h) → citrulline → ornithine →
creol red (pH Enterobacter yellow putrescine
indicator) aerogenes—
yellow to purple Arginine—
Escherichia
Arginine— coli—yellow
Pseudomonas
aeruginosa—
yellow to purple
44 Phenol red YELLOW ** All enterobacteriaciae ferment glucose and
CHO carbohydrates lactose fermentation is used to differentiate
COLOR
fermentati or CTA sugars groups of genera within the family
on with phenol
red indicators
are most
commonly
used