Microbiology Lab Final

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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


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