Interpreteing
Biochemical Media
BIOL250
Dr. Barbara R. Heard
Mannitol Salt Agar
• Reported relative to Growth/Fermentation
• +/+ = Growth and fermentation = organism can grow in
hypertonic condition created by salt, and can ferment
mannitol (top left section)
• +/- = Growth, no fermentation = organism can grow in
hypertonic condition created by salt, but cannot ferment
mannitol (bottom section)
• -/- = No growth or fermentation = organism cannot grow in
hypertonic condition created by salt. If there is no growth,
there cannot be fermentation (right section)
https://quizlet.com/204869548/medical-microbiology-media-flash-cards/
Eosin Methylene Blue
• Reported relative to Growth/Fermentation
• +/+ = Growth and fermentation = organism can grow in presence
of methylene blue (likely Gram negative) and can ferment lactose
and/or sucrose. Eosin turns from colorless to black in acidic pH;
dark purple and green metallic sheen = lactose and/or sucrose
fermentation; brown-pink color = slow lactose fermentation (top
two sections)
• +/- = Growth, no fermentation = organism can grow in presence of
methylene blue (likely Gram negative), but cannot ferment lactose
and/or sucrose (bottom section)
• -/- = No growth or fermentation = organism cannot grow in
presence of methylene blue (likely Gram positive). If there is no
growth, there cannot be fermentation (not pictured)
https://www.youtube.com/watch?v=DYDDwE-HhCM
Starch Agar
• Polysaccharide starch composed of glucose molecules bonded
together. Organisms producing enzymes alpha amylase or oligo-
1,6 glucosidase break down starch to glucose for energy
• After incubation, iodine added to agar. Look for change in color of
agar next to growth
• Starch + iodine = dark blue/black color = negative (organism could
not break down starch – no production of those enzymes)
• Glucose + iodine = amber color = positive (organism produced
enzyme(s) and broke down starch)
https://cwoer.ccbcmd.edu/science/microbiology/Lab%20Manual/lab8/posstar.html
Blood Agar
• If organism can produce a hemolysin, can completely or partially
lyse RBCs
• Beta (β) hemolysis = complete lysis of RBCs = Organism is beta
hemolytic
• Alpha (⍺) hemolysis = partial lysis of RBCs = Organism is alpha
hemolytic
• Gamma (γ) hemolysis = no lysis of RBCs = Organism is gamma
hemolytic
https://microbeonline.com/blood-agar-composition-preparation-uses-and-types-of-hemolysis/
Phenol Red with
Carbohydrate
• Fermentation produces acidic end products
• Phenol red turns yellow in acidic pH
• Some organisms produce CO2 with fermentation; some do
not
• Reported as Negative, A, AG, (some labs report -/-, A/-, A/G)
https://microbiologyinfo.com/phenol-red-fermentation-test/
MRVP
• Methyl Red test – After addition of methyl red indicator, test read
immediately. Cherry red = positive
• Voges Proskauer test – After addition of reagents, one hour must
pass before reporting negative result. Cherry red = positive
• Reported as MR result/VP result: +/+, +/-, -/+, -/-
https://microbiologynote.com/methyl-red-and-voges-proskauer-test-principle-result/
TSI Interpretation
Reported As What is Learned?
Tube A A/A+G Ferments glucose & lactose &/or sucrose; gas
Tube B A/A Ferments glucose & lactose &/or sucrose
Tube C K/A Ferments glucose
Tube D K/A+H2S Ferments glucose; enzymes to reduce sulfur
Tube E Uninoculated
A = Acid production from fermentation
K = Alkaline (pink) from protein degradation
Other possibilities:
A/H2S – Ferments glucose and lactose and/or sucrose; produces H2S
K/NC - No fermentation
Motility Test Medium
• Lateral growth away from central stab line indicates motility
• Flagella make motility possible
• Reported as Motile or Non-motile
https://www.facebook.com/elmicrobiologistee/photos/a.115577503249004/123218095818278/
Urea Broth
• Some bacteria can hydrolyze urea by producing exoenzyme
urease produces end products ammonia and carbon
dioxide alkaline pH
• Phenol red indicator turns fuchsia in alkaline conditions =
positive result
https://asm.org/getattachment/ac4fe214-106d-407c-b6c6-e3bb49ac6ffb/urease-test-protocol-3223.pdf
Thioglycollate Broth
• Organism’s aerotolerance determined by where in broth the
organism can grow
• Pink coloration indicates high oxygen levels
https://microbenotes.com/thioglycollate-broth/
https://quizlet.com/231666408/lab-midterm-fluid-thioglycollate-medium-flash-cards/
Catalase Test
• Growth on NAS used for test
• Some organisms produce enzyme catalase which catalyzes
H2O2 H2O and O2
• Production of bubbles indicates organism is catalase positive
https://asm.org/Protocols/Catalase-Test-Protocol
