Media Biochemical Test
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107 Terms
Which Staphylococcus identification tests are used
BAP, MSA, Mstaph broth, Coagulase
How is the Indole test performed
Inoculate Tryptone broth with inoculating loop
What does the Indole test detect
Ability to produce indole
What enzyme is involved in indole production
Tryptophanase
What amino acid is broken down in the Indole test
Tryptophan
What medium is used for the Indole test
Tryptone broth contains tryptophan
What reagent is added in the Indole test
Kovac’s reagent
What are the components of Kovac’s reagent
Hydrochloric acid, dimethylaminobenzaldehyde, and amyl alcohol
What is the original color of Kovac’s reagent
Yellow
What indicates a positive Indole test
Red color at the top part of the test tube
How are the MR and VP tests performed
Inoculate 2 glucose broths with inoculating loop, add MR to one tube and VP reagents to the other after 48 hours
What family do MR and VP tests differentiate
Enterobacteriaceae
What does the Methyl Red test detect
Acid end products from glucose fermentation
What does the Voges Proskauer test detect
Acetoin production from glucose fermentation
What medium is used in MR/VP tests
Glucose broth
What indicator is used in the MR test
Methyl Red
What is VP reagent A
5% Alpha-Naphthol and ethanol
What is VP reagent B
Potassium Hydroxide and Deionized Water
What indicates a positive MR test
Red indicating pH below 6
What indicates a negative MR test
Yellow indicating no acid production
What indicates a positive VP test
Red after VP reagents are added
What indicates a negative VP test
No color change
What does the Citrate test determine
Ability to consume citrate as sole carbon source and ammonium as sole nitrogen source
What family is differentiated by the Citrate test
Enterobacteriaceae
What medium is used in the Citrate test
Simmon’s Citrate Agar
What is the carbon source in Citrate agar
Sodium citrate
What is the nitrogen source in Citrate agar
Ammonium ion
What pH indicator is used in Citrate agar
Bromthymol blue
What indicates a positive Citrate test
Blue
What indicates a negative Citrate test
Green
How is the H2S test in SIM performed
Stab SIM media with inoculating needle
What does the H2S test determine
Ability to reduce sulfur into H2S
What family is differentiated by the H2S test
Enterobacteriaceae
What medium is used in the H2S test
SIM media
What sulfur-containing amino acid is present in SIM
Cysteine
What additional components are in SIM media
Sodium thiosulfate and peptonized iron or ferrous sulfate
What indicates a positive H2S test
Black precipitate present
What indicates a negative H2S test
No black precipitate
How is the Urea hydrolysis test performed
Inoculate Urea broth with inoculating loop
What does the Urea test determine
Ability to hydrolyze urea to make ammonia using urease
What enzyme is detected in the Urea test
Urease
What medium is used in the Urea test
Urea broth
What are the components of Urea broth
Yeast extract, monopotassium phosphate, disodium phosphate, urea, phenol red indicator
What is the original color of Urea broth
Yellow-orange
What indicates a positive Urea test
Bright pink color
What indicates a negative Urea test
No color change and no ammonia is made
How is the Motility test performed
Stab motility media with inoculating needle
What does the Motility test determine
Ability of bacteria to be motile
What medium is used in the Motility test
Motility media
What are the components of Motility media
Tryptose, sodium chloride, agar, and a color indicator
What indicates a positive Motility test
Growth going out away from the stab line
What indicates a negative Motility test
Growth only along the stab line
How is the Lactose fermentation test performed
Inoculate lactose broth with inoculating loop
What does the Lactose fermentation test determine
Ability to ferment lactose
What medium is used for Lactose fermentation
Lactose broth
What are the components of Lactose broth
Beef extract, gelatin peptone, and lactose
What indicator is used in the Lactose test
Phenol red
What indicates a positive Lactose fermentation test
Yellow
What indicates a negative Lactose fermentation test
No color change or reddish
How is the Sucrose fermentation test performed
Inoculate sucrose broth with inoculating loop
What does the Sucrose test determine
Ability to ferment sucrose and produce acid end-product
What family is differentiated by the Sucrose test
Enterobacteriaceae
What medium is used in the Sucrose test
Sucrose broth
What are the components of Sucrose broth
Beef extract, gelatin peptone, and sucrose
What indicator is used in the Sucrose test
Phenol red
What indicates a positive Sucrose fermentation test
Yellow
What indicates a negative Sucrose fermentation test
No color change or reddish
How is the Glucose fermentation and gas test performed
Inoculate broth with inoculating loop
What does the Glucose test determine
Ability to ferment glucose and produce gas and/or acid end-product
What family is differentiated by the Glucose test
Enterobacteriaceae
What medium is used in the Glucose test
Glucose broth
What are the components of Glucose broth
Beef extract, gelatine peptone, and glucose
What indicator is used in the Glucose test
Phenol red
What is used to detect gas production
Durham tube
What indicates acid production in Glucose fermentation
Yellow
What indicates gas production in Glucose fermentation
Bubble in the Durham tube
What indicates a completely negative Glucose test
No color change or reddish color and no bubble
How is the BAP with Novobiocin test performed
Inoculate BAP plate using streak plate method and place novobiocin disk in first quadrant
How long is the BAP test incubated
24-48 hours
What does the BAP with Novobiocin test determine
Resistance or sensitivity to Novobiocin and hemolysis
What medium is used in the BAP test
Nutrient agar with 5% sheep's blood
What antibiotic disk is used in the BAP test
Novobiocin
What indicates hemolysis on BAP
Media surrounding colonies turns translucent or green
How is Novobiocin resistance determined
Zone of clearing smaller than manufacturer’s recommendation
What result indicates Novobiocin resistance and sensitivity
Top streak R for NB and bottom S
What type of bacteria lyse red blood cells
Hemolytic bacteria
What type of bacteria do not lyse red blood cells
Non-hemolytic bacteria
How is the Mannitol Salt Agar test performed
Inoculate MSA plate using streak plate method and incubate 24-48 hours
What does MSA test for
Ability to tolerate 7.5% salt concentration and ferment mannitol
Why is MSA selective
Selects for salt tolerant bacteria
Why is MSA differential
Differentiates organisms by mannitol fermentation
What are the components of MSA
Nutrient agar, mannitol, 7% sodium chloride, phenol red
What indicates salt tolerance on MSA
Growth
What indicates no salt tolerance on MSA
No growth
What indicates positive mannitol fermentation on MSA
Yellow zones around colonies
What indicates negative mannitol fermentation on MSA
Media remains pink
How is the Mstaph broth test performed
Inoculate broth and incubate 24-48 hours
What does Mstaph broth test for
Ability to tolerate 7.5% salt concentration and ferment mannitol
Why is Mstaph broth selective
Selects for salt tolerant Staphylococcus
What does Mstaph broth contain
Nutrients appropriate for growing Staphylococcus and 7.5% salt