Alkaline Peptone Broth or Alkaline Peptone Water: enrichment medium Ior increasing the

numbers oI target bacteria that can multiply under alkaline conditions (e.g., Jibrio cholerae,
while inhibiting the growth oI contaminating Ilora that cannot multiply at an elevated pH
Bile Esculin Agar (BEA): selective medium Ior the detection oI Iecal streptococci (group D
and enterococci; tests ability oI the organism to hydrolyze esculin to esculetin. Brownish-
black colonies surrounded by a black zone are positive.
Oxgall (bile is inhibitory agent.
Ferric citrate is indicator.
Blood Agar (BAP): consists oI a basal medium such as TSA enriched with 5 deIibrinated
sheep blood or in some locations, horse blood. This is the most commonly used medium, and
supports the growth oI most oI the common Iastidious organisms, as well as, all oI the less
Iastidious organisms (e.g., coliIorms.
BAP will be used to practice streaking technique to obtain isolated colonies; to observe
diIIerences in colony morphology; to determine various patterns oI hemolytic activity; to
perIorm CAMP test Ior Group B streptococci.
Brilliant Green Agar: selective medium Ior the isolation oI $almonella, except $ typhi.
Brilliant green is the selective agent.
Phenol red is the pH indicator.
ampy-blood agar (ampy-BAP): Brucella agar supplemented with sheep blood and
vancomycin (inhibits Gram-positives, trimethoprim (broad spectrum , and cephalothin
(inhibits streptococci
ary-Blair Transport Medium: semi-solid alkaline medium used to transport sensitive
specimens to the laboratory when the specimen cannot be cultured immediately
hopped (ooked) Meat Medium: used Ior cultivation and maintenance oI lostridium and
to evaluate proteolysis.
hocolate Agar: blood agar prepared by heating blood to 56
o
C until medium becomes
brown or chocolate in color. Heating the blood releases both X and V growth Iactors and also
destroys the inhibitors oI V Iactor. These Iactors are required Ior the growth oI most species
oI aemophilus and also Neisseria gonorrhoeae.
For clinical gonococcal specimens, isolates are subcultured Irom the selective primary
medium to a noninhibitory enrichment medium, e.g., chocolate agar supplemented with 1
IsoVitaleX to obtain a pure culture oI the isolate.
oagulase Test Medium: citrated rabbit plasma which clots in the presence oI the enzyme
coagulase.
ystine (ysteine) Tellurite Blood Agar: both a diIIerential and selective medium Ior the
isolation oI diphtheriae; however, a Iew strains oI streptococci and staphylococci are able
to grow on this medium; diphtheriae (also $taphylococcus produces gray to black
colonies because the tellurite is reduced intracellularly to tellurium
ystine Trypticase Agar (TA): carbohydrate-supplemented CTA medium dispensed in
tubes is used to detect Iermentation oI the various carbohydrates and can be used Ior
detemination oI motility.
Phenol red is pH indicator.
ystine Trypticase Agar (TA) Sugar Fermentation Media: CTA supplemented with
either glucose, maltose or sucrose; tests Ior utilization oI carbohydrates.
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Glucose Broth: Ior anaerobic Iermentation (overlaid with mineral oil aIter inoculation.
Phenol red is the pH indicator.
Loeffler (Loeffler's) Medium: primary isolation medium Ior orynebacterium diphtheriae;
distinctive colonial morphology is observed
Lysine Decarboxylase Medium: a yellow color indicates a low pH and that the test is
negative (Iailure to produce an amine by decarboxylation oI lysine.
Bromcresol purple is the pH indicator.
Maconkey Agar: an inhibitory and diIIerential medium used to distinguish lactose-
Iermenting Gram-negative organisms Irom nonIermenters.
Crystal violet, bile salts and neutral red are inhibitory agents.
Neutral red is the pH indicator.
Mannitol Salt Agar (MSA): Ior selective isolation oI coagulase-positive, mannitol-
Iermenting $taphylococcus. Mannitol Iermentation by pathogenic staphylococci is indicated
by a yellow halo surrounding the colonies.
Sodium chloride is the inhibitory agent.
Phenol red is the pH indicator.
Motility-Indole-Ornithine Agar: motility is indicated by the character oI the growth in the
butt oI this tube. Motile organisms will produce a general clouding oI the medium or a Iuzzy
stab line. Non-motile organisms will give a sharply delineated stab line. The ornithine
reaction is indicated by the color in the butt oI the tube. Yellow indicates a negative test
(Iailure to decarboxylate ornithine; purple is a positive test(decarboxylation oI ornithine.
Kovac's reagent is added to the tube to determine the indole reaction. Red indicates a positive
reaction (indol production; yellow is a negative test (Iailure to produce indol Irom
tryptophan.
Mueller Hinton Agar: a rich medium consisting oI 30 beeI inIusion, 1.75 acidicase
peptone, 0.15 starch and 1.7 agar that supports the growth oI most microorganisms. It is
commonly used Ior antibiotic susceptibility testing: disk diIIusion antibiotic susceptibility;
antibiotic serum level measurements; MBC determination.
Mueller-Hinton Broth: culture medium Ior broth tube antibiotic MIC assay
Return to Top of Media List
itrate Broth: some bacteria (e.g., !seudomonas aeruginosa have respiratory enzyme
systems that can use nitrate as a terminal electron acceptor. The product oI the reaction is
nitrite. Some oI the organisms that reduce nitrate to nitrite will then reduce the nitrite Iurther.
In the scheme below, Iirst test Ior nitrite by a colorimetric test. II this test is negative, it can
mean either that nitrite was not reduced, or that it was reduced beyond the nitrite stage. This
can be resolved by the addition oI zinc dust; iI nitrate is still present, the zinc will reduce it
chemically to nitrite, which will then by revealed by the colorimetric reaction.
Procedure: To the nitrate broth, aIter 48 hours oI incubation, add 0.2 ml oI acid reagent
(Solution A, a mixture oI acetic acid and sulIanilic acid, and then 0.2 ml oI dimethyl-alpha-
napthylamine reagent (Solution B. II nitrite is present you will get a red color: this is a
positive test Ior nitrate reduction. II there is no color, pick up some zinc dust on the end oI an
applicator stick, and add it to the tube. ZINC DUST SUSPENDED IN AIR CAN BE
EXPLOSIVE; KEEP AWAY FROM FLAMES! II you get a red color at this stage, what can
you conclude? What iI no color is obtained?
utrient Agar: contains 0.5 gelysate peptone, 0.3 beeI extract, and 1.5 agar, and will
support the growth oI many organisms which are not nutritionally Iastidious (e.g.,
staphylococci, and enterics. (Note: Agar is a substance which melts at 100
o
C and solidiIies
at about 42
o
C; it has no nutritional beneIits, but is only a stabilizer to allow Ior solidiIication
oI the medium.
OF Glucose Medium: to detect Iermentation or oxidation oI glucose (see above.
Bromthymol blue is pH indicator.
OF Maltose Medium: to detect Iermentation or oxidation oI maltose (see above.
Bromthymol blue is pH.
Phenylethyl Alcohol Agar (PEA): Ior the isolation oI $taphylococcus and inhibition oI
Gram-negative bacilli (particularly !roteus.
Phenylethyl alcohol is the inhibitory agent.
Salmonella - Shigella (SS) Agar: isolation and diIIerential medium Ior pathogenic Gram-
negative enteric bacilli, in particular, $almonella and $higella. Inhibitory Ior coliIorms.
Serum Tellurite Agar: isolation medium Ior orynebacterium diphtheriae; diphtheriae
(also $taphylococcus produces gray to black colonies because the tellurite is reduced
intracellularly to tellurium
SF Broth: ($treptococcus Enterococcus( faecalis broth: selective medium Ior the detection
oI Iecal streptococci (group D and enterococci Irom water, milk and other materials oI
sanitary importance. Growth oI all other cocci is inhibited. Fermentation oI glucose is
indicated by a color change in the broth.
Sodium azide is the inhibitory agent.
Bromcresol purple is the indicator.
Simmons itrate Agar: utilization oI citrate as the sole source oI carbon is indicated by the
medium turning a deep blue color because oI an alkaline reaction. Non-utilizers will leave the
green color oI the slant unchanged.
Bromthymol blue is the indicator.
Skirrow's agar: peptone and soy protein base agar supplemented with lysed horse blood and
vancomycin (inhibits Gram-positives, polymyxin B (antiIungal, and trimethoprim (broad
spectrum
Sugar Utilization Medium: supplemented with glucose (with Durham tubes to determine
gas production, sucrose, mannitol, or lactose.
Phenol red is the pH indicator.
Return to Top of Media List
TBS Agar (ThiosulIate-itrate-Bile salts-Sucrose agar: DiIIerential and selective plating
medium Ior Jibrio adjusted to pH 8.6 (alkaline; Jibrio cholerae colonies appear yellow;
other Jibrio spp. colonies appear green
Thayer-Martin Agar: chocolate agar supplemented with antibiotic inhibitors Ior selective
isolation oI pathogenic Neisseria
Vancomycin inhibits Gram-positive organisms.
Colistin inhibits Gram-positive enteric organisms.
Nystatin inhibits yeast.
Triple Sugar Iron Agar (TSI): this is a key medium Ior use in beginning the identiIication
oI a Gram-negative bacilli oI the enteric group. It contains glucose (0.1, lactose (1,
sucrose (1 and peptone (2as nutritional sources. Sodium thiosulIate serves as the
electron receptor Ior reduction oI sulIur and production oI H
2
S. Detects Iermentation oI
sucrose, lacotse, glucose, as well as production oI hydrogen sulIide and/or gas.
Phenol red is pH indicator; Ierric ammonium citrate is H
2
S indicator.
Click here to view Explanation oI TSI Reactions.
Trypticase Lactose Agar: to determine motility and lactose Iermentation by anaerobes.
Phenol red is pH indicator.
Trypticase itrate Broth: to determine indole production and nitrate reduction.
Trypticase Salicin Agar: to determine motility and salicin Iermentation by anaerobes.
Phenol red is pH indicator.
Trypticase Soy Agar (TSA): enriched medium containing 1.5 trypticase peptone, 0.5
phytone peptones, 0.5 NaCl, 1.5 agar that supports the growth oI many oI the more
Iastidious organisms: e.g. streptococci, and some members oI the genera Neisseria, Brucella,
orynebacterium, Listeria, !asteurella, Jibrio, Erysipelothrix, etc.
To be used to practice streaking technique and to observe diIIerences in colony
morphology, as well as, Ior recovery oI microorganisms Irom skin; to perIorm catalase test;
to test requirements Ior X and/or V Iactors.
Trypticase Sucrose Agar: to determine motility and sucrose Iermentation by anaerobes.
Phenol red is pH indicator.
Urease Broth or Urea Agar Slant: Prompt hydrolysis oI urea by !roteus species is
indicated by a deep pink color appearing in the medium within eight hours. At 18 hours, this
color will have spread throughout the whole tube. Many strains oI lebsiella, Enterobacter
and itrobacter will yield a positive reaction, but usually the pink color will be limited to the
slant in 24 to 48 hours. Do not reincubate tubes that show any evidence oI color change.
LD Agar (Xylose lysine desoxycholate agar: Selective media Ior the isolation oI $higella
and other enteric pathogens.
LT-4 Agar: Selective media Ior the isolation oI $almonella developed at and patented by
investigators at the University oI Maryland and the USDA. It contains peptone, yeast extract,
lysine, lactose, sucrose, xylose as nutritional sources. Sodium thiosulIate acts as electron
receptor Ior reduction oI sulIur to H
2
S. The medium is highly inhibitory Ior non-salmonellae.
Tergitol-4 also may be inhibitory to $ typhi and $ choleraesuis.
Tergitol-4 (surIactant is the inhibitory agent.
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