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What does TSA stand for?
Tryptic Soy Agar
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What test is associated with TSA media?
Several (it is a general purpose media)
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Is TSA selective or differential?
Neither, it is general purpose media
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What is the pH indicator in TSA?
Doesn't have one
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What does TSA grow?
Just about any bacteria, but not fastidious ones
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What does TSB stand for?
Tryptic Soy Broth
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What test is TSB associated with?
None in particular (it is general purpose media)
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Is TSB selective or differential?
Neither, it is general purpose
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What is the pH indicator in TSB?
Doesn't have one
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What does TSB grow?
Just about any bacteria, but not fastidious ones
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What does SAB stand for?
Sabouraud agar
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What test is SAB used for?
Not associated with any particular test
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What does SAB grow?
Fungi
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How does SAB prevent growth of bacteria on it?
It is slightly acidic (pH = 5.6)
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Is SAB selective or differential?
Selective (grows fungi, but excludes bacteria)
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What is the pH indicator in SAB?
Doesn't have one
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What does BAP stand for?
Blood Agar Plate
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How is BAP made?
Add 5% sheep blood to TSA
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What test is BAP used for?
Production of hemolysins
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Is BAP differential or selective?
Differential (differentiates pathogenic species of Staph and Strep from non-pathogenic species)
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What is the pH indicator in BAP?
Doesn't have one
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How is a positive test indicated in BAP?
- Alpha hemolysin production will show a brown/green halo around the colonies
- Beta hemolysin production will show a colorless halo around the colonies
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What reagents do you add after incubating BAP?
None
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What is thioglycollate broth used for?
Grows anaerobes by binding free O2 in the broth to create an anaerobic environment
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Is thioglycollate broth selective or differential?
Differential (what it differentiates is not indicated)
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What is the pH indicator in thioglycollate broth?
Doesn't have one
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What test is fermentation broth used for?
Fermentation of CHO
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What CHO are used in fermentation tests?
Sugars (glucose, sucrose, lactose, mannitol)
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What is the pH indicator in fermentation broth?
Phenol red
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How is a positive test result indicated in fermentation broth?
The indicator will change from red to yellow
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What causes the positive result in fermentation broth?
Production of acidic by-products from fermentation of CHO
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What reagents are added after incubation of fermentation broth?
None
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How is 7% NaCl agar made?
Add 7% NaCl to TSA
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Is 7% NaCl agar selective or differential?
Selective (for G+ cocci)
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What is the pH indicator in 7% NaCl agar?
Doesn't have one
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What test is 7% NaCl agar used for?
Not associated with any test, it is used as a selective culture media
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What does EMB stand for?
Eosin Methylene Blue agar
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Is EMB selective or differential?
Both
- -- Selects for G- bugs
- -- Differentiates lactose fermenters vs. non-fermenters
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What is the pH indicator in EMB?
Methylene Blue
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How is a positive test result indicated in EMB?
Lactose fermenters will be black (or black-centered) with a colorless or pink margin
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What reagents are added to EMB after incubation?
None
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What test is Simmon's citrate agar used for?
Citrate test (part of the IMViC series)
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What color is Simmon's citrate agar prior to inoculation?
Green
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What is the pH indicator in Simmon's citrate agar?
Bromthymol blue
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What is the Simmon's citrate agar testing for?
The ability to use sodium citrate as a source of C
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How is a positive test indicated in Simmon's citrate agar?
Color change from green to blue
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What reagents do you add to Simmon's citrate agar after incubation?
None
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Is Simmon's citrate agar selective or differential?
Both (but most often used to differentiate Enterobacteriaceae)
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What are the two kinds of Pseudomonas agars?
P and F (pyocyanin and flourescein)
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Why are there two kinds of Pseudomonas agars?
Each type enhances the production of one pigment and inhibits the other pigment
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Are the Pseudomonas agars selective or differential?
Differential, they distinguish sub-species of P. aeruginosa based on pigment production
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What test are the Pseudomonas agars used for?
Not test in particular, they are used to run down sub-species
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What does OF basal media stand for?
Oxidation/Fermentation basal media
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What test is OF basal media used in?
Oxidation/Fermentation test
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What does OF basal media also contain?
CHO (glucose in our case)
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What is the pH indicator in OF basal media?
Bromthymol blue
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How is a positive result indicated in the OF test?
- Positive for oxidation if the color changes to yellow in ONLY the open tube (the one without oil in it)
- Positive for fermentation if BOTH tubes are yellow
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What reagents are added to the OF basal media after incubation?
None
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What is the purpose of the OF test?
To differentiate nonenteric G- bugs from enterics
- -- Enterics are facultative anaerobes that will ferment glucose (turn both tubes yellow)
- -- Pseudomonas are aerobes that oxidize glucose (turn the open tube yellow)
- -- Alcaligenes faecalis will do nothing to the tubes
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What test is phenylalanine agar used for?
Phenylalanine test
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Is phenylalanine agar selective or differential?
Differential (distinguishes Proteus and Providencia from other Enterobacteriaceae)
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What pH indicator is used in phenylalanine agar?
None
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How is a positive test indicated in phenylalanine agar?
Formation of a green color
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What reagents are added to the phenylalanine agar after incubation?
FeCl3 [iron (III) chloride or ferric chloride]
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What does TSI stand for?
Triple Sugar Iron agar
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What test is TSI used for?
Production of H2S
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Is TSI selective or differential?
Differential (distinguishes members of the Enterobacteriaceae family)
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What is the pH indicator in TSI?
Phenol red
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How is a positive test result indicated in TSI?
- Black color = H2S production
- Red slant = - for fermentation of CHO
- Yellow slant = + for fermentation of CHO
- Bubbles/tears in slant = CO2 production
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What reagents are added to TSI after incubation?
None
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What are the 3 sugars in TSI?
Sucrose, glucose, and lactose
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What test is decarboxylation broth used for?
Decarboxylation of AA
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Is decarboxylation broth selective or differential?
Differential (helps distinguish members of the Enterobacteriaceae)
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What pH indicator is used in decarboxylation broth?
Bromcresol purple
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What AA are in decarboxylation broth?
Lysine, ornithine, and arginine
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What reagents are added to decarboxylation broth after incubation?
None
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How is a positive test result indicated in decarboxylation broth?
The broth will turn purple and become turbid
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What test is starch agar used for?
Starch Hydrolysis test
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Is starch agar selective or differential?
Differential (distinguishes Enterobacteriaceae)
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What is the pH indicator in starch agar?
Doesn't have one
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How is a positive test indicated in starch agar?
A cleared halo around the growth (indicating the hydrolysis of the starch)
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What reagents are added to the starch agar after incubation?
Dilute iodine sol'n
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What test is skim milk agar used for?
Casein Hydrolysis test
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How do you make skim milk agar?
Add sterile skim milk to TSA
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Is skim milk agar selective or differential?
Differential (distinguishes Enterobacteriaceae)
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What is the pH indicator in skim milk agar?
Doesn't have one
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How is a positive test result indicated in skim milk agar?
Cleared area (zone of hydrolysis) around a colony
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What reagents are added to skim milk agar following incubation?
None
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What test is DNAase agar used for?
DNAase test
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Is DNAase agar selective or differential?
Both
- -- Selects for G+ Staph and G- bacilli
- -- Distinguishes both pathogenic Staph from non-pathogenic and Serratia marcesens from other G- bacilli
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What is the pH indicator in DNAase agar?
Doesn't have one
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How is a positive test result determined in DNAase agar?
Clearing of the methyl green dye surrounding the growth
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Why is the DNAase test used to distinguish pathogenic Staph (why does it work)?
The production of DNAase exoenzyme strongly correlates to coagulase production (coagulase is characteristic to Staphylococcus aureus)
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What reagents are added to DNAase agar after incubation?
None
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What is the purpose of the DNAase test?
The primary purpose is to distinguish pathogenic Staph from non-pathogenic, but it can also be used to distinguish Serratia marcesens from other G- bacilli
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What test is tryptone broth used for?
Indole production test (part of the IMViC series)
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Is tryptone broth selective or differential?
Differential (distinguishes Enterobacteriaceae)
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What pH indicator is used in tryptone broth?
Doesn't have one
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How is a positive test result determined in tryptone broth?
Formation of a red color
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What reagents are added to tryptone broth after incubation?
Kovac's sol'n
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What does MR/VP stand for?
- MR = Methyl Red test
- VP = Voges-Proskauer test
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What test is MR/VP broth used for?
The MR test and the VP test (part of the IMViC series)
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Is the MR/VP broth selective or differential?
Differential (distinguishes Escherichia coli from Enterobacter aerogenes)
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What is the pH indicator used in the MR/VP broth?
Methyl red
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What is the CHO in MR/VP broth?
Glucose
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If your MR test is positive, what would you expect your VP test to be?
Negative (they should result in opposing values)
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How is a positive test result determined in the MR/VP broth?
- MR+ = the color will stay red
- VP+ = development of a pink or red color
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What reagents are added to MR/VP broth after incubation?
- MR: add 5 drops of Methyl red indicator
- VP: add 15 drops of Barritt's reagent A and 5 drops of Barritt's reagent B
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What does MSA stand for?
Mannitol Salt Agar
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What test is MSA used for?
No particular test, it is a growth medium
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Is MSA selective or differential?
Both
- -- Selective for G+ cocci
- -- Differentiates pathogenic Staph from non-pathogenic
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What pH indicator is used in MSA?
Doesn't have one
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What test is Nitrate broth used for?
Nitrate Reduction test
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Is the nitrate broth selective or differential?
Differential (distinguishes Enterobacteriaceae)
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What pH indicator is used in the nitrate broth?
Doesn't have one
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How is a positive test result indicated in nitrate broth?
- Production of a red color after adding reagents A&B (indicates + for nitrite)
- Production of a brown/yellow color after adding Nessler's reagent (indicates + for ammonia)
- Look for bubbles in the Durham tube
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What reagents are added to the nitrate broth after incubation?
- First, add Nitrate Reagent A and Reagent B to test for nitrite
- Second, add Nessler's reagent to test for ammonia production (we didn't do this)
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What test is Mueller-Hinton agar used for?
The Kirby-Bauer antibiotic evaluation method
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Is Mueller-Hinton agar selective or differential?
Neither, it is simply a growth medium
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How is a positive test result determined on Mueller-Hinton agar?
There will be a zone of inhibition surrounding the antibiotic discs (which you measure the diameter of to determine the effectiveness of the agent)
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What test is malonate broth used for?
Malonate Utilization test
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Is malonate broth selective or differential?
Differential (distinguishes Enterobacteriaceae)
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What is the pH indicator used in malonate broth?
Bromthymol blue
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How is a positive test result determined in malonate broth?
- Blue color = bug used the malonate
- Yellow color = bug did not use the malonate but did ferment the glucose
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What reagents are added to malonate broth after incubation?
None
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What materials are used in the oxidase test?
Oxistrips (strips of filter paper soaked in Kovac's sol'n)
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How is a positive test result determined in the oxidase test?
Production of a blue color on the oxistrip or the swab
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Is the oxidase test selective or differential?
Neither (because you only test one bug at a time)
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What does a positive oxidase test indicate?
That your tested bug is a Pseudomonas
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What does a negative oxidase test indicate?
That your tested bug is an Enterobacteria
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What is the purpose of the oxidase test?
To differentiate members of Family Pseudomonadaceae from members of Family Enterobacteriaceae
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What reagent is used in the catalase test?
30% H2O2
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How is a positive test result determined in a catalase test?
Formation of bubbles
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What is the purpose of the catalase test?
Distinguishes Streptococcus (-) from Micrococcus (+) and Bacillus (+) from Clostridium (-)
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Is the catalase test selective or differential?
Neither (since you only test one bug at a time)
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What materials are used to perform a coagulase test?
Citrated rabbit plasma (rabbit plasma with sodium citrate added to prevent coagulation)
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How is a positive test result determined in the coagulase test?
Clotting of the plasma
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What does a positive coagulase test result indicate?
Pathogenic Staphylococcus (S. aureus)
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What does a negative coagulase test result indicate?
Non-pathogenic Staphylococcus (S. epidermidis)
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Give an example of a G+ bacillus.
Anything from the following genera:
- Bacillus
- Clostridium
- Mycobacterium
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Give an example of a G- bacillus.
Anything from the following genera:
- Klebsiella
- Brucella
- Escherichia
- Bordetella
- Proteus
- Legionella
- Pseudomonas
- Yersinia
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Give an example of a G+ coccus.
Anything from the following genera:
- Streptococcus
- Staphylococcus
- Micrococcus
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Give an example of a G- coccus
Anything from the following genus:
Neisseria
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Give an example of a bug + for capsules
Klebsiella pneumoniae
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Give an example of a bug + for endospores
Clostridium perfringens
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Give an example of a bug + for acid-fast staining.
Mycobacterium tuberculosis
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