Microbiology Lab Final
Oxidase Test Differential Agent
Chromogenic reducing agent
Simmon's Citrate Test Differential Agent
Citrate
Bacitracin Susceptibility Test Positive Results
Clearing > 10mm (indicates organism is susceptible to bacitracin)
Motility (SIM) Test Positive Results
Cloudiness that often obscures the stab line
Coagulase Test Indicator
Coagulation (Clotting)
Nitrate reductase
Converts NO3 to NO2
Purpose of adding Mineral Oil to previously inoculated media
Creates an anaerobic environment, promoting fermentation (creating an acidic environment. The acidic environment causes lysine decarboxylase-positive microbes to make the enzyme, the resulting alkaline products reverse the reaction causing the media to turn purple)
Catalase Test Indicator
Creation of oxygen bubbles (indicates presence of catalase)
Kligler Iron Agar Used For
Differentiate organisms based on their ability to ferment glucose only, glucose AND lactose, and their ability to reduce sulfur
Bile Esculin Test Used For
Enterococci, Group D streptococci (inhibits all other gram + bacteria)
Bile Esculin Test Differential Agent
Esculin (streptococci and enterococci hydrolyze esculin to esculetin)
Phenylalanine Deaminase Test Indicator
Ferric Chloride (FeCl3 added after incubation)
Bile Esculin Test Indicator
Ferric Citrate
Coagulase Test Differential Agent
Fibrinogen (in blood plasma)
Coagulase Test Positive Results
Fluid becomes thick
Gelatin Hydrolysis Test Differential Agent
Gelatin
Phenol Red Glucose Test Differential Agent
Glucose (fermentation of glucose requires presence of enzymes necessary to metabolize them)
Sulfur (SIM) Test Indicator
H2S forms a black percipitate
Blood Agar Indicator
Hemolytic Activity
Phenylalanine Deaminase Test
Identifies presence phenylalanine deaminase (enzyme that removes the amino group from phenylalanine)
Oxidase Test Purple Result
Oxidase Test Positive (oxidase enzyme is present)
VP Test Reddish Results
Oxidizing of Acetoin to Diacetyl
Urea Test Used to Dertermine
The microbes ability to produce urease
Vogues-Proskauer Test Detectable Intermediate
Acetoin
MR-(VP) Test Differential Agent
(MR) Glucose (fermentation leads to acid production)
Ornithine Decarboxylase Test Purple Results
(Ornithine) pH above 6.8 (+)
Ornithine Decarboxylase Test Yellow Results
(Ornithine) pH below 5.2 ( - )
Phenol Red Sucrose Test Primary Indicator
(Sucrose ) Phenol Red
Phenol Red Sucrose Test Secondary Indicator
(Sucrose) Durham Tube
Urea Test Reddish-pink Results
(Urea Test) pH above 8.4 (+)
Urea Test Orange Results
(Urea Test) pH below 8.4 (-)
(MR)-VP Test Differential Agent
(VP) Glucose
Phenol Red Glucose Test Secondary Indicator
(glucose) Durham Tube
Phenol Red Glucose Test Primary Indicator
(glucose) Phenol Red
Lysine Decarboxylase Test Purple Results
(lysine) pH above 6.8 (+)
Lysine Decarboxylase Test Yellow Results
(lysine) pH below 5.2 (-)
Phenol Red Xylose Secondary Indicator
(xylose) Durham Tube
Phenol Red Xylose Test Primary Indicator
(xylose) Phenol Red
Kligler Iron Agar Differential Agents
1 % lactose, 0.1% glucose and sodium thiosulfate
Undetectable Alcohol Vogues-Proskauer tests for
2, 3-butanediol
Motility (SIM) Test Differential Agent
3.5% Agar
Blood Agar Differential Agent
5 % Blood
What is Cytochrome c oxidase?
A carrier within the ETC of some bacteria
Lysine Decarboxylase Test Used to Determine
Accumulation of alkaline products
Is ammonia alkaline or acidic?
Alkaline
Urea Test reacts to Alkaline or Acidic Conditions
Alkaline (when metabolized, urea produces ammonia which causes the pH of the media to increase)
Microbes that survive will convert ammonium phosphate to what?
Ammonia
What is catalase?
An enzyme that converts hydrogen peroxide to water and oxygen
Purpose of Zinc in Nitrate Reduction
Artificially converts NO3 to NO2
Bile Esculin Test Acronym
BESC
Kligler Iron Agar Yellow THROUGHOUT
BOTH glucose AND lactose was fermented
Bacitracin Susceptibility Test Differential Agent
Bacitracin (antibiotic)
Blood Agar Hemolytic Activity Results
Beta Hemolysis (complete degradation), Alpha Hemolysis (incomplete degradation), Gamma Hemolysis (no degradation)
Bile Esculin Test Selective Agent
Bile
Bile Esculin Test No color change result
Bile Esculin Negative Result
Bile Esculin Test Black color change result
Bile Esculin Positive Result (ferric citrate reacts with esculetin)
Blood Agar Differs from Columbia CNA Blood Agar
Blood Agar lacks the Antibiotics in CNA for selectivity
Lysine Decarboxylase Test Indicator
Bromcresol Purple
Ornithine Decarboxylase Test Indicator
Bromcresol Purple
Simmon's Citrate Test Indicator
Bromthymol Blue
MR-VP Test Media Type
Broth (one single media, divided into 2 portions)
Bacitracin Susceptibility Test
Indicates if the antibiotic Bacitracin in effective against cell wall formation
I in SIM Test
Indole
Indole (SIM) Test Red Results
Indole positive
Kligler Iron Agar Acronym
KIA
Kligler Iron Agar Red Results
Kligler Negative Results (alkaline conditions; no fermentation has occurred)
Kligler Iron Agar Yellow Results
Kligler Positive Results (Presence of Acid; fermentation has occurred)
Kligler Iron Agar Black Presence Results
Kligler Positive Results for H2S (iron turns black in presence of H2S)
Indole (SIM) Test Indicator
Kovac's Reagent (dimethylaminobenzaldehyde, HCL)
Lysine Decarboxylase Test Differential Agent
Lysine
Methyl Red / Vogues-Proskauer Tests Acronym
MR-VP
MR-(VP) Test Methyl Red Reagent Indicator
Methyl Red Dye
Gelatin Hydrolysis Test Used to Determine
Microbes that secrete gelatinase (to catabolize gelatin and absorb the amino acids)
M in SIM Test
Motility
Products of removing the amino group from phenylalanine
NH3 and phenylpyruvic acid
Nitrate Reduction Test Differential Agent
Nitrate
Nitrate Reduction Test Primary Indicators
Nitrate I Reagent (Dimethyl-alpha-naphthylamine), Nitrate II Reagent (Sulfanilic Acid)
Nitrate Reduction Reagent
Nitrate I Reagent (Dimethyl-alpha-naphthylamine; ONLY reacts with NO2)
Nitrate Reduction Test Secondary Indicator Red Result
Nitrate Reductase Negative (Darker Red)
Nitrate Reduction Test Primary Indicator Red Result
Nitrate Reductase Positive
Nitrate Reduction Test Secondary Indicator Clear Result
Nitrate Reductase Positive
Kligler Iron Agar Yellow Butt ONLY
ONLY glucose was fermented
Indole (SIM) Test Used to Determine
Organims that can produce tryptophanase
Motility (SIM) Test Used to Determine
Organisms that are motile via flagella
Coagulase Test Used to Determine
Organisms that can bind fibrinogen
Sulfur (SIM) Test Used to Determine
Organisms that can produce H2S (hydrogen sulfide) during metabolism
Catalase Test Used to Determine
Organisms that produce Catalase
Ornithine Decarboxylase Test Differential Agent
Ornithine
Urea Test Indicator
Phenol Red
Phenol Red Glucose Test Bubble in Durham Tube Results
Phenol Red Glucose Durham Tube Positive Results (fermentation releases gas, creating a bubble in the tube)
Phenol Red Glucose Test Yellow Results
Phenol Red Glucose Positive Results (fermentation creates acid)
Phenol Red Sucrose Bubble in Durham Tube Results
Phenol Red Sucrose Durham Tube Positive Results (fermentation releases gas, creating the bubble)
Phenol Red Sucrose Test Yellow Results
Phenol Red Sucrose Positive results (fermentation creates acid, causing color change)
Phenol Red Xylose Yellow results
Phenol Red Xylose positive results (fermentation creates acid, causing color change)
Kligler Iron Agar Indicators
Phenol Red and Ferric Ammonium Sulfate
Phenylalanine Deaminase Test Differential Agent
Phenylalanine
Phenylalanine Deaminase Test No Color Change Result (yellow)
Phenylalanine Deaminase Negative result
Phenylalanine Deaminase Test Green Result
Phenylalanine Deaminase Positive result (ferric chloride reacts directly with phenylpyruvic acid)
Nitrate Reduction Test Bubble formation in Durham Tube
Positive result for N2 gas creation
Oxidase Test Used to Determine
Presence of Cytochrome c oxidase
Sulfur Indole Motility Test Acronym
SIM
Kligler Iron Agar Media Type
Slanted
Sulfur (SIM) Test Differential Agent
Sodium Thiosulfate
Blood Agar Used to Determine
Staphylococcus, Streptococcus, Enterococcus (from less virulent)
Phenol Red Sucrose Test Differential Agent
Sucrose (fermentation of sucrose requires presence of enzymes necessary to metabolize them)
Kligler Iron Agar Used to Determine
Sugar fermenters and/or H2S producers
S in SIM Test
Sulfur
Denitrification
The conversion of nitrate to nitrogen gas (NO3 to NH3 or N2 gas)
Indole (SIM) Test Differential Agent
Tryptophan
Indole (SIM) Test Positive Results Indicator
Tryptophan is catabolized to indole
Motility (SIM) Test Indicator
Turbidity
Nitrate Reduction Test Primary Indicator Clear Result
Unknown (must use secondary indicator Zinc)
Urea Test Differential Agent
Urea
Vogues-Proskauer Reagent A Indicator
VP Reagent A (alpha-Naphthol)
Vogues-Proskauer Reagent B Indicator
VP Reagent B (KOH -- Potassium hydroxide)
Methyl Red Test Used to Determine
Whether end product of glucose fermentation are mixed acids
Vogues-Proskauer Test Used to Determine
Whether end product of glucose fermentation is alcohol
Phenol Red Xylose Test Differential Agent
Xylose (fermentation of xylose requires presence of enzymes necessary to metabolize them)
Nitrate Reduction Test Secondary Indicator
Zinc
Bacitracin Susceptibility Test Indicator
Zone of Clearing
When is Kovac's Reagent added to the media
after incubation
Only microbes that make citrate-permease can convert what?
citrate to pyruvate
Lysine Decarboxylase and Ornithine Decarboxylase are...
enzymes that remove the carboxyl group from peptone (Decarboxylation results in accumulation of alkaline products)
Kligler Iron Agar Inoculation Techinques
fish tail AND stab
Gelatin Hydrolysis Test Indicator
liquification of gelatin
Durham Tubes used for
notification of gas production (fermentation)
MR-(VP) Test Methyl Red Reagent Red Results
pH 5.5 or less (indicates presence of mixed acids)
MR-(VP) Test Methyl Red Reagent Yellow-orange Results
pH 6.2
Simmon's Citrate Test Blue Result
pH 7.6 (ammonia released raises pH) (+)
How Kligler Iron Agar Media is divided
slant and butt
How to read Kligler Iron Agar Results
slant/ butt/ gas/ H2S