Term
| What does the Tryptophanase/Indole test testing for? |
|
Definition
| The ability of an organism to hydrolyze tryptophan into indole |
|
|
Term
| What type of media is used in the Trypophanase/Indole test? What does it contain? |
|
Definition
| SIM agar which contains tryptophan |
|
|
Term
| What reagent is used to detect indole in the Tryptophanase test? |
|
Definition
|
|
Term
| Is tryptophanase an endo or exo enzyme? |
|
Definition
|
|
Term
| What are the 4 tests in the IMViC series? |
|
Definition
1. Indole/Tryptophanase
2. Citrase
3. Methyl Red
4. Voges-Proskauer |
|
|
Term
| What does a positive Indole test look like? |
|
Definition
| Red reagent layer after adding Kovak's |
|
|
Term
| What does a negative Indole test look like? |
|
Definition
| No reaction after addition of Kovak's. No color. |
|
|
Term
| What does the Citrase test test for? |
|
Definition
| Citrase converts citrate into oxaloacetic acid and acetate which is converted into pyruvic acid and CO2 (alkaline) |
|
|
Term
| What does a positive citrase test look like? |
|
Definition
| blue media (from alkaline) |
|
|
Term
| What does a negative citrus test look like? |
|
Definition
| green media (no change in pH) |
|
|
Term
| What does the Methyl Red test for? What is this detected by? |
|
Definition
| Production of acidic end products from breakdown of glucose (detected by pH indicator Methyl Red) |
|
|
Term
| What class of organisms does the Methyl Red test target and why? |
|
Definition
| enteric bacteria because glucose is a major substrate used by this class. |
|
|
Term
| What does a positive Methyl Red test look like? what pH range is this for? |
|
Definition
|
|
Term
| What does a negative Methyl Red test look like? what pH range is this for? |
|
Definition
|
|
Term
| What does Voges-Proskauer test for? (include broad trait and specific end-product) |
|
Definition
| neutral end products of glucose breakdown- acetylmethylarbinol |
|
|
Term
| What reagent is used in the Voges-Proskauer test? |
|
Definition
| Barritt's reagent A and B |
|
|
Term
| What does a positive Voges-Proskauer test look like? |
|
Definition
| deep rose color after 15 minutes with Barritt's agent added |
|
|
Term
| What does a negative Voges-Proskauer test look like? |
|
Definition
|
|
Term
| What additional traits is the Indole media used to test? |
|
Definition
| H2S production and motility |
|
|
Term
| What does the Hydrogen Sulfide test test for? |
|
Definition
| breakdown of Sulfur containing amino acids and other inorganic molecules to create H2S |
|
|
Term
| What media is used to detect H2S |
|
Definition
|
|
Term
| What does a positive H2S test look like? |
|
Definition
|
|
Term
| How do you detect motility in a bacteria? what media is used for this? |
|
Definition
| when culture growth is not limited to the line of inoculation (SIM agar) |
|
|
Term
| What enzyme is being detected in the Urease test? What are the substrates and products in the reaction and what is the products pH? |
|
Definition
Enzyme is urease
breaks N and C bonds in amides to produce ammonia (alkaline) |
|
|
Term
| What are the two reagents in the Urease test? |
|
Definition
| Urea (amide compound) and phenol red (pH indicator) |
|
|
Term
| What pH does phenyl red test for in the Urease test? |
|
Definition
|
|
Term
| What does a positive urease test look like? |
|
Definition
|
|
Term
| what does a negative urease test look like? |
|
Definition
|
|
Term
| What does the Catalase test test for? |
|
Definition
| presence of catalase which breaks down hydrogen peroxide |
|
|
Term
| What is the function of catalase breaking down hydrogen peroxide? where does this hydrogen peroxide come from? |
|
Definition
| Hydrogen peroxide is a byproduct of superoxide dismutase breaking down toxic superoxides |
|
|
Term
| What does a positive catalase test look like? |
|
Definition
| bubble production when hydrogen peroxide is added to culture (O2 bubbles) |
|
|
Term
| What does the oxidase test test for? |
|
Definition
| Cytochrome oxidase removal of electrons of final protein of the ETC by O2. |
|
|
Term
| what reagent is used in the oxidase test? |
|
Definition
| oxidase strip with oxidase reagent |
|
|
Term
| what does a positive oxidase reaction look like? |
|
Definition
| dark purple color in 10-30 seconds |
|
|
Term
| what does ELISA stand for? |
|
Definition
| enzyme-linked immunosorbent assay |
|
|
Term
| What does a direct ELISA test for? |
|
Definition
|
|
Term
| what does an indirect ELISA test for? |
|
Definition
|
|
Term
| What is the binding order for Direct ELISA? |
|
Definition
| antigen--> known antibody with fluorescent label--> observe under light |
|
|
Term
| What is the binding order for Indirect ELISA? |
|
Definition
| antigen bound to slide--> patient serum added (antibody)-->florescent anti-antibody (antibody sandwich) |
|
|
Term
| For carbohydrate fermentation, what media and pH indicator were used to identify fermentation? |
|
Definition
| media was nutrient broth with phenol red |
|
|
Term
| what were the three sugars used to measure carbohydrate fermentation? |
|
Definition
| lactose, glucose, sucrose |
|
|
Term
| Which bacteria had acid and gas productions (yellow with bubble)? |
|
Definition
|
|
Term
| Which bacteria had no fermentation (no acid or gas)? |
|
Definition
| A. faecalis and Pseudomonas |
|
|
Term
|
Definition
| selects among a specific groups of bacteria- usually share common trait (gram positive, gram negative, halophiles) |
|
|
Term
| 3 types of selective Media |
|
Definition
-Crystal Violet Agar
-Phenylethyl Alcohol Agar
-7.5% Sodium Chloride Agar |
|
|
Term
Phenylethyl Alcohol Agar
(what does it inhibit, what does it select for) |
|
Definition
Inhibits Gram negative (alcohol disrupts membrane)
Selects for Gram Positive cocci |
|
|
Term
Crystal Violet Agar
(what does it inhibit, what does it select for) |
|
Definition
Inhibits Gram Positive
Selects for Gram Negative |
|
|
Term
7.5% Sodium Chloride Agar
(what does it inhibit, what class of bacteria does it select for, give an example of a bacteria in this class) |
|
Definition
inhibits anything non-halophilic
Selects for halophiles
Ex: Staphylococcus |
|
|
Term
|
Definition
| Selects specific organism within a group of bacteria (identifies species) |
|
|
Term
| 3 types of Differential Media |
|
Definition
-Mannitol Salt Agar
-MacConkey Agar
-Eosin-Methylene Blue Agar |
|
|
Term
Mannitol Salt Agar
(What group does it select for, what does it differentiate and how, provide examples) |
|
Definition
salt selects for halophiles- Staphylococcus
Mannitol fermentation (pathogenic trait) differentiates epidermidis from aureus
Ex: S. aureus ferments mannitol and turns red media yellow |
|
|
Term
MacConkey Agar
(What group does it select for, what does it differentiate and how, provide examples) |
|
Definition
Selects for gram negative (crystal violet inhibits gram positive)
Differentiates lactose fermenting bacteria (coliforms)
Lactose fermentation produces acids which turn the pH indicator red
Ex: E. coli, citrobacter, Enterobacter, Klebsiella |
|
|
Term
Eosin-Methylene Blue Agar
(What group does it select for, what does it differentiate and how, provide examples) |
|
Definition
Selects for gram negative lactose fermenters (coliforms)
Differentiates between E. coli and all other coliforms- E. coli produces metallic green color
-Enterobacter aerogenes (coliforms)- pink colonies w central dark purple spot |
|
|
Term
|
Definition
Contain nutrients to support a wide variety of bacterial growth as well as additional unique requirements of fastidious growers
Ex: blood agar |
|
|
Term
Blood Agar
(what does it differentiate? what are the three types of growth?) |
|
Definition
Differentiates hemolytic Streptococcus species (enriched and differential media)
1. Alpha hemolysis
2. Beta hemolysis
3. Gamma hemolysis |
|
|
Term
Alpha hemolysis
(what does it look like and what organism shows this type of growth?) |
|
Definition
Partial breakdown of hemoglobin shows green/light brown sheen around colonies (biliverden)
Ex. E. coli |
|
|
Term
Betta hemolysis
(what does it look like and what organism shows this type of growth?) |
|
Definition
Full breakdown of red blood cells leaves clear space- lyse cells using beta-hemolysin (streptolycin)
Ex: S. pyogenes |
|
|
Term
Gamma hemolysis
(what does it look like and what organism shows this type of growth?) |
|
Definition
no hemolysis- whitish grey sheen on surface of media
Ex: S. epidermidis |
|
|
Term
| Enteric Bacteria (2 types) |
|
Definition
Bacteria that colonize the intestinal tract- coliform and non-coliform
-ferment glucose
-facultative anaerobes
-usually gram negative bacilli |
|
|
Term
|
Definition
lactose fermenting enterics
Ex: E. coli, citrobacter, Enterobacter, Klebsiella |
|
|
Term
|
Definition
|
|
Term
|
Definition
| Alpha Hemolysis (Blood Agar) |
|
|
Term
|
Definition
| Betta Hemolysis (Blood Agar) |
|
|
Term
|
Definition
| Gamma Hemolysis (Blood Agar) |
|
|
Term
|
Definition
|
|
Term
|
Definition
| Eosin Methylene Blue Agar (E. coli) |
|
|
Term
|
Definition
| Eosin Methylene Blue Agar (Enterobacter Aerogenes) |
|
|
Term
|
Definition
|
|
Term
|
Definition
|
|
Term
|
Definition
|
|
Term
|
Definition
|
|
Term
| Best temp for S. marcenscense |
|
Definition
| 20 degrees C (mesophil)- red pigment |
|
|
Term
| 3 toxic Oxygen forms (superoxides) |
|
Definition
Superoxide
Peroxide
Hydroxyl |
|
|
Term
| 3 enzymes for superoxide breakdown. Where are they found? |
|
Definition
Found in aerobes
- Superoxide dismutase
-Catalase
-Peroxidase |
|
|
Term
| What agar medium was used in the oxygen requirement experiments? |
|
Definition
| Brain heart fusions in deep tubes |
|
|
Term
| What area in the agar do aerobes grow? |
|
Definition
| at the surface of the tube |
|
|
Term
| what area in the agar do obligate anaerobes grow? |
|
Definition
|
|
Term
| what area in the agar do facultative anaerobes grow? |
|
Definition
| throughout the agar with more at the surface |
|
|
Term
| What is the final electron acceptor for aerobes? |
|
Definition
|
|
Term
| what is the final elector acceptor for obligate anaerobes? |
|
Definition
| molecules other than oxygen |
|
|
Term
| In the catalase experiment, what solution is added to the tubes |
|
Definition
|
|
Term
| What represents a positive reaction in the catalase test? |
|
Definition
|
|
Term
| in a normal cell, where would the hydrogen peroxide being neutralized by catalase come from? |
|
Definition
| superoxide dismutase breaks down toxic superoxides and produces hydrogen peroxide. |
|
|
Term
| What agar and reagent is used in the amylase test? |
|
Definition
| starch agar and Gram's iodine |
|
|
Term
| What reaction is being tested for in the amylase test? |
|
Definition
| Starch hydrolysis into simple sugars (glucose) |
|
|
Term
| What does a positive Amylase test looks like (with Gram's iodine added)? |
|
Definition
| cleared zone (halo) surrounding the growth |
|
|
Term
| What does a negative Amylase test looks like (with Gram's iodine added)? |
|
Definition
| Blue to black color in the medium |
|
|
Term
| What agar is used in the lipase test? |
|
Definition
| Tributyrin agar (with lipid)- blue color |
|
|
Term
| What reaction is being tested in the lipase test? |
|
Definition
| hydrolysis of ester bonds by lipase |
|
|
Term
| What does a positive lipase test look like? |
|
Definition
| clear halo surrounding growth. Loss of apacity |
|
|
Term
| What does a negative lipase test look like? |
|
Definition
| Medium remains opaque blue color surrounding growth |
|
|
Term
| What media is used in the Casein test? |
|
Definition
|
|
Term
| What reaction is being tested in the Casein test? |
|
Definition
| breakdown of casein, a protein in milk, by protease |
|
|
Term
| What does a positive results in the Casein test look like? |
|
Definition
| loss of opacity surrounding the bacterial growth |
|
|
Term
| What does a negative result in the casein test look like? |
|
Definition
| medium surrounding the bacterial growth remains opaque |
|
|
Term
|
Definition
|
|
Term
|
Definition
|
|
Term
|
Definition
|
|
Term
|
Definition
|
|
Term
| What agar and broth was used for the temperature experiment? |
|
Definition
| Nutrient broth and TSA platse |
|
|
Term
| What method was used to inoculate the oxygen requirement bacteria in brain hear fusion? |
|
Definition
|
|
Term
| What are the 4 exoenzymes? |
|
Definition
1. lipase
2. amylase
3. gelatin
4. casein |
|
|
Term
| in the gelatin experiment, what enzyme is breaking down gelatin and what is it breaking gelatin down into? |
|
Definition
| gelatinase breaks gelatin down into amino acids |
|
|
Term
| how many ml to one micro liter? |
|
Definition
|
|
Term
| In the DNA extraction experiment, what is done to the strawberry cells to expose the DNA? |
|
Definition
| The cells are treated with detergent (SDS) in a buffer solution |
|
|
Term
| In the DNA extraction experiment, how is protein precipitated out of the solution? |
|
Definition
| presence of salt in the DNA extraction buffer |
|
|
Term
| In the DNA extraction experiment, how is cellular waste and particles removed from solution? |
|
Definition
|
|
Term
| In the DNA extraction experiment, how is DNA precipitated from the filtered solution? |
|
Definition
|
|
Term
| Transformation- What two genes were present in the plasmid vector? |
|
Definition
|
|
Term
| Transformation- how did we select for transformed bacteria? |
|
Definition
|
|
Term
| Transformation- what was done to E.coli to induce competency? |
|
Definition
|
|
Term
| Transformation- what were the growth areas surrounding transformed bacteria called? |
|
Definition
|
|
Term
| What is another name for the disk diffusion test (antibiotics)? |
|
Definition
|
|
Term
| What is the area surrounding an antibiotic disk that restricts growth called? |
|
Definition
|
|
Term
| What are two typical gram + bacteria? |
|
Definition
|
|
Term
| What three sugars do phenol red tubes contain for carbohydrate fermentation? |
|
Definition
1. glucose
2. lactose (galactose + glucose)
3. sucrose (fructose + glucose) |
|
|
Term
| In carbohydrate fermentation, which bacteria usually produces acid? |
|
Definition
|
|
Term
| In carbohydrate fermentation, which bacteria doesn't undergo gas or acid production? |
|
Definition
|
|
Term
| What type of agar is used to test for dental caries susceptibility? |
|
Definition
|
|
Term
| What does a Snyder test test for? |
|
Definition
| acid production from the breakdown of glucose by lactobacilli |
|
|
Term
| What pH does Snyder agar test for? What is the color change for a positive result? |
|
Definition
| test for acidic environment. Changes from green to yellow in presence of pH 4.4 and below |
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|
