Exam 1 (Ch 9 - Biochemical Tests, etc.) Flashcards
What is the general procedure for the catalase test?
- Place bacterial colony on glass slide
- Add drop of 3.0% H2O2
- Check for bubbles
What is the positive result of the catalase test?
Give an example positive organism.
Positive result of catalase test:
- Bubbles
Example:
- Staphylococcus and Micrococcus species
What is the negative result of the catalase test?
Give an example negative organism.
Negative result of catalase test:
- Absence of bubbles
Example:
- Streptococcus species
Enzyme which catalyzes the conversion of fibrinogen to fibrin
Coagulase
2 forms of the coagulase test
- Tube method
2. Commercial latex agglutination test
What substance is used in the tube method of the coagulase test?
Rabbit plasma containing EDTA
What is the positive result of the coagulase test?
Give an example positive organism.
Positive result of coagulase test:
- Coagulation/agglutination
Example:
- Staphylococcus aureus
What is the negative result of the coagulase test?
Give an example negative organism.
Negative result of coagulase test:
- Absence of coagulation/agglutination
Example:
- Staphylococcus epidermis
A term in which the organism produces clumping factor and/or protein A
Agglutination
What is the function of the spot indole test?
- Detects indole production, based on organism possessing tryptophanase
- Smear colony across white filter paper saturated with Kovac’s or Ehrlich’s test reagent.
If using the spot indole test, from what 2 types of agar can colonies be?
- SBA agar
2. CHOC agar
What 2 test reagents are used in the spot indole test?
- Kovac’s
2. Ehrlich’s
What is the positive result of the spot indole test?
Give an example positive organism.
Positive result of spot indole test:
- Appearance of a purple color
Example:
- E. coli
What is the negative result of the spot indole test?
Give an example negative organism.
Negative result of spot indole test:
- Absence of a purple color
Example:
- Pseudomonas aeruginosa
What kind of organisms is the oxidase test used for?
Gram negative organisms
What enzyme is used in the oxidase test?
1% cytochrome C oxidase
What is the function of 1% cytochrome C oxidase?
Facilitates transfer of electrons to oxygen in aerobic respiration
What is the general procedure for the oxidase test?
- Use a wooden stick to smear a colony across filter paper or a swab saturated with oxidase test reagent.
- Do not use a needle or metal loop, as they can cause a false positive
What is the positive result of the oxidase test?
Give an example positive organism.
Positive result of oxidase test:
- Appearance of a purple color within 10-30 seconds
Example:
- Neisseria species
- Pseudomonas aeruginosa
- Campylobacter species
What is the negative result of the oxidase test?
Give an example negative organism.
Negative result of oxidase test:
- No appearance of a purple color within 10-30 seconds
Example:
- E. coli
What kind of organisms is the modified oxidase test used for?
Gram positive organisms
What enzyme is used in the modified oxidase test?
6% cytochrome C oxidase
What is the purpose of the modified oxidase test?
It is used to aid in identification of catalase-positive Gram Positive Cocci
Give an example positive organism of the modified oxidase test
Micrococcus species
Give an example negative organism of the modified oxidase test
Staphylococcus species
What is the purpose of the esculin hydrolysis test?
- Differentiation and ID of Gram positive, catalase negative cocci
- Tests the ability to hydrolyze esculin
What is the general procedure for the esculin hydrolysis test?
- Use paper with esculin embedded in it
- Smear organism on paper
- Incubate 15 minutes
What is the positive result of the esculin hydrolysis test?
Give an example positive organism.
Positive result of esculin hydrolysis test:
- Black color (under UV light)
Example:
- Enterococcus species
What is the negative result of the esculin hydrolysis test?
Give an example negative organism.
Negative result of esculin hydrolysis test:
- White color (under UV light)
Example:
- Group B Streptococcus (S. agalactiae)
- Viridans streptococci
Generally, PYR hydrolysis test is used in differentiation and ID of what organisms?
Gram-positive, catalase-negative cocci
The PYR hydrolysis test is used to presumptively identify what 2 sets of species?
- Enterococcus species
2. Streptococcus pyogenes
What is the ridiculously named agent used in the PYR hydrolysis test?
L-pyrrolidonyl-β-naphthylamide
What is the positive result of the PYR hydrolysis test?
Give an example positive organism.
Positive result of PYR hydrolysis test:
- Presence of red color
Example:
- Enterococcus species
- Streptococcus pyogenes
What is the negative result of the PYR hydrolysis test?
Give an example negative organism.
Negative result of PYR hydrolysis test:
- Absence of red color
Example:
- Group B Streptococcus (S. agalactiae)
- E. coli
The bile solubility test is used in the presumptive ID of what organism?
Streptococcus pneumoniae
What is the positive result of the bile solubility test?
Give an example positive organism.
Positive result of bile solubility test:
- If the colony lyses in the presence of the bile salt solution within 30 minutes, it is a positive result
- Clear = positive
Example:
- Streptococcus pneumoniae
What is the negative result of the bile solubility test?
Give an example negative organism.
Negative result of bile solubility test:
- If the colony does NOT lyse in the presence of the bile salt solution within 30 minutes, it is a negative result
- Cloudy = negative
Example:
- Streptococcus mitis
What is the purpose of the carbohydrate fermentation broths?
It tests the utilization of carbohydrates as an energy source in an anaerobic environment.
In carbohydrate fermentation broths:
What is the negative reaction?
No color change (stays red-orange)
In carbohydrate fermentation broths:
What is the positive reaction?
Yellow color
In carbohydrate fermentation broths:
What is the positive reaction with gas production?
Yellow color and bubbles
What is the purpose of the oxidative-fermentative test?
It tests the utilization of carbohydrates, by fermentation or by oxidative pathway.
What is the general procedure for the oxidative-fermentative test?
Two tubes are inoculated for each carbohydrate tested
- One gets overlay of mineral oil to produce anaerobic condition
- The other gets left as oxygenic
In oxidative-fermentative tests:
Why does one tube have an anaerobic condition, whereas the other is left as oxygenic?
- Fermenters can produce acid in both tubes
# Can ferment with or without oxygen - Oxidizers only produce acid in the oxygenic tube
The oxidative-fermentative test can be used to differentiate what species?
It can be used to differentiate Enterobacteriaceae (fermenters) from other gram-negative bacilli (nonfermenters like Pseudomonas species).
What is the TSI test primarily used to differentiate?
It is primarily used to differentiate Enterobacteriaceae species.
What kind of agar is used in the TSI test? How is it used in the procedure?
Slant agar
- Consists of top (slant) and bottom (butt or deep)
- Lid must be left loose
- Inoculate the agar with needle, then streak the slant
In the TSI test:
What do the red, yellow, and black colors indicate?
If bubbles are present, what does that indicate?
- Red = alkaline
- Yellow = acid
- Black = H2S production
- Bubbles = gas production
In the TSI test:
What does red/yellow mean?
Give an example of an organism that shows this reaction.
- Alkaline/Acid (K/A)
- Glucose only fermenter
- Shigella sonnei
In the TSI test:
What does yellow/yellow mean?
Give an example of an organism that shows this reaction.
- Acid/Acid (A/A)
- Glucose and lactose or sucrose fermenter
- E. coli
In the TSI test:
What does red/red mean?
Give an example of an organism that shows this reaction.
- Alkaline/Alkaline (K/K)
- Nonfermenter
- Pseudomonas aeruginosa
In the TSI test:
What does red/no change mean?
- Alkaline/no change (K/NC)
- Nonfermenter
In the TSI test:
What does no change/no change mean?
- NC/NC
- Nonfermenter
- Verify that there is growth on the slant
In the TSI test:
What does the black butt mean?
- H2S production
- Assume acid reaction if the black covers up the color in the agar
In the TSI test:
Give an example of an organism that shows a red/black color.
Salmonella typhi
In the TSI test:
Give an example of an organism that shows a yellow/black color.
Proteus mirabilis
What is the purpose of the KIA test?
- Same as TSI, but no sucrose present
- More sensitive for H2S production
What does the LIA test differentiate and screen for?
- Differentiate among the Enterobacteriaceae species
- Screens for Salmonella and Shigella species
What all does the LIA test detect?
- Decarboxylation of lysine
- Deamination of lysine
- H2S production
What happens in the procedure for the LIA test?
- Inoculate slant
- Stab butt twice