Lab Exam 3 Gram - Flashcards
You should know a few of the potential genera used in the unknown experiment.
- Escherichia
- Pseudomonas
- Citrobacter
For each test, you should know what a positive test result looks like and what a negative test result looks like.
What color is the Simmons’ slant before it is inoculated?
Green
The Simmons’ slant contains bromothymol blue. In this case, the pH indicator will not turn yellow. What color is a Simmons’ slant that is positive for nutrient utilization?
Blue
What color is the tube before it is incubated? Simmons slant test.
Green
What is the carbon source in the Simmons’ slant?
Citrate
What is the nitrogen source in the Simmons’ slant?
Ammonia
How does nutrient utilization affect the pH of the Simmons’ slant?
Increases pH, making it more alkaline
Litmus Milk
Litmus milk broth contains two key ingredients, skim milk and litmus powder. What are the two functions of the litmus powder in the litmus milk broth?
1.) pH indicator
2.) Litmus can be reduced
Litmus Milk
What are the three reactions that can occur with the litmus powder?
1.) Purple = negative for fermentation (alkaline)
2.) Pink = positive for fermentation (acidic)
3.) White = reduction (can see white at bottom of tube)
Litmus Milk
Each of the litmus reactions produce a different color. Match each of the three litmus reactions to pink, purple, or white.
1.) Purple = negative for fermentation (alkaline)
2.) Pink = positive for fermentation (acidic)
3.) White = reduction (can see white at bottom of tube)
Litmus Milk
What are the two possible reactions that can occur with the milk protein in the litmus milk broth?
1.) pH indicator
2.) Litmus can be reduced
Litmus Milk
What is peptonization?
Enzymes breakdown proteins into peptides
Litmus Milk
How will coagulation affect the broth?
Broth solidifies
Litmus Milk
What causes the broth to coagulate?
Too much acid release into media
SIM stab
What does the abbreviation ‘SIM’ stand for?
- Sulfide
- Indole
- Motility
SIM stab
Which of the three tests did we not record data for?
Motility
SIM stab
Which amino acid is degraded to produce ‘S’?
Cysteine
SIM stab
What is the enzyme that is responsible for ‘S’?
Cysteine desulfurase.
SIM stab
What are the three products that are produced as part of ‘S’?
Hydrogen sulfide (H2S), pyruvic acid, and ammonia
SIM stab
What are the three products that are produced as part of ‘S’?
Hydrogen sulfide (H2S), pyruvic acid, and ammonia.
SIM stab
Which specific product of the three is detected by ‘S’?
Hydrogen sulfide (H2S)
SIM stab
What color is a positive ‘S’ test?
Black precipitate
SIM stab
Which amino acid is degraded to produce ‘I’?
Tryptophan
SIM stab
What is the enzyme that is responsible for ‘I’?
Tryptophanase
SIM stab
What are the three products that are produced as part of ‘I’?
Indole, pyruvic acid, and ammonia
SIM stab
Which specific product of the three is detected by ‘I’?
Indole
SIM stab
What reagent is used to test for ‘I’?
Kovac’s reagent
What color is a positive ‘I’ test?
Red ring at the top of tube
MRVP
What does the abbreviation ‘MRVP’ stand for?
Methyl Red Voges-Proskauer
MRVP
Which reagent tests for the production of mixed acids by fermentation?
Methyl red reagent
MRVP
How does the color of the reagent that tests for mixed acid fermentation change with decreasing pH?
Turns red with decreasing pH
MRVP
How many days does the 2,3-butanediol test using an MRVP tube incubate for?
5 days
MRVP/2,3-butanediol
Why doesn’t 2,3-butanediol react with a pH indicator, like phenol red?
Because alcohol doesn’t change the pH, therefore the pH indicator doesn’t react.
Vogues Proskaver
What is the exact procedure for performing the VP test in lab?
- Label empty test tube for our unknown
- Pipette 1 ml of MRVP into the unknown tube
- Add 18 drops of reagent A and 18 drops of reagent B to the unknown tube
- Shake tube every 10 minutes
- After 30 minutes, check color
Vogues Proskaver
What is the purpose of shaking the small tube?
We shake to oxidize acetoin into diacetyl (if its’s present)
Vogues Proskaver
Which final chemical product will be created in the VP test?
Diacetyl
Vogues Proskaver
How long does the VP test sit before the color change is complete?
30 Minutes
What color is a positive VP test? What color is a negative test result?
- Positive: Red
- Negative: Brown
Brilliant Green Agar
What kills Gram + & some Gram – in the Brilliant Green Agar? (how is it selective?)
Brilliant green dye selects against gram + bacteria, and some gram - bacteria.
Brilliant Green Agar
What sugars are found in the Brilliant Green?
Hint: Two of them
- Sucrose
- Lactose
Brilliant Green Agar
What is the pH indicator for the Brilliant green agar?
Phenol red
Brilliant Green Agar
What color is brilliant green agar that is positive for fermentation (Acidic)?
Lime green
Brilliant Green Agar
What color is brilliant green agar for a nonfermenter (Basic)?
Bright pink
Brilliant Green Agar
What color is brilliant green agar when the bacteria is dead (negative)?
Orangish-Brown since the plate remains the same color
Catalase Test
What type of media was used for the catalase test?
Hydrogen peroxide (H2O2)
Catalase Test
You should be able to write out the complete catalase reaction, including the names and molecular formulas of all substrates and products.
Catalase reaction: 2 H2O2 → 2 H2O + O2
* Substrate: Hydrogen Peroxide
* Product: Water and Oxygen
Briefly describe the appearance of a positive catalase test.
Bubble formation
Which product of catalase activity is responsible for the visible reaction?
Oxygen gas
What is the protein cytochrome oxidase normally used for? Where specifically is cytochrome oxidase located?
Cytochrome oxidase is involved in the electron transport chain. It is located in the bacterial cell membrane.
What is the exact reaction of cytochrome oxidase?
The reduction of cytochrome c using oxygen as the final electron acceptor
What color is a positive cytochrome oxidase test?
Dark purple
What type of media was used for the oxidase test?
TSA plate streaked with the unknown on one half and Pseudomonas aeruginosa on the other half.
What is in the Hektoen agar that makes it selective?
Bile
Hektoen Agar
What three carbohydrates does it contain?
- Lactose
- Sucrose
- Salicin
Hektoen Agar
What are the pH indicators?
- Bromthymol Blue
- Andrades Acid
Hektoen Agar
What is the fermentation color change (acidic)?
Turns Bright Orange
Hektoen Agar
What is the non-fermenter color change (basic)?
Blue-Green