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microbio > Practicum II > Flashcards

Practicum II Flashcards

Lab Diagnostic Tests

1
Q

Hydrolysis of Starch

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Medium: Starch agar plate
  • Substrate: Starch (Polysaccharide)
  • Enzyme/Pathway: Hydrolysis of Amylase
  • pH Ind./Reagents: Iodine
  • End Products: Maltose (Disacharide)
  • (+) appearance: Light/Golden area around colonies
  • (-) appearance: Dark around colonies
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2
Q

Hydrolysis of Gelatin

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Medium: Nutrient gelatin tall
  • Substrate: gelatin (protein)
  • Enzyme/Pathway: Hydrolysis of gelatinase
  • pH Ind./Reagents: N/A
  • End Products: amino acids
  • (+) appearance: Liquid
  • (-) appearance: Solid
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3
Q

Fermentation of Carbohydrates

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Medium: CHO Broth (1 CHO, Basal Medium, Phenol Red, Durham Tube)
  • Substrate: 1x CHO (eg. glucose)
  • Enzyme/Pathway: Fermentation
  • pH Ind./Reagents: N/A
  • End Products: A: ATP + Organic Acid, AG: ATP + Organic Acid + CO2 gas, SA: ATP + Organic Acid
  • (+) appearance: A: Yellow, AG: Yellow + Bubble. SA: Orange
  • (-) appearance: -: Red, B: Magenta, SB: half red/half magenta
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4
Q

Fermentation

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Medium: Litmus Milk
  • Substrate: Lactose
  • Enzyme/Pathway: Fermentation
  • pH Ind./Reagents: N/A
  • End Products: A: Organic Acid + ATP, AC: Acid + ATP + Hard Curd (Protein), AC: Acid + ATP+ Hard Curd + CO2 gas
  • (+) appearance: A: Pink milk, AC: Pink milk + Curd, AC: Pink milk + curd with gas cracks
  • (-) appearance: Purple milk like control, blueberry shake with no solid
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5
Q

Alkanization

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Medium: Litmus Milk
  • Substrate: Casein
  • Enzyme/Pathway: Alkanization
  • pH Ind./Reagents: N/A
  • End Products: Polypeptides (colloid) + basic amines
  • (+) appearance: B: Blue milk (blueberry shake)
  • (-) appearance: Purple milk (like control), pink milk
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6
Q

Peptonization

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Medium: Litmus Milk
  • Substrate: Casein & Lactalbumin (colloid) / Proteins
  • Enzyme/Pathway: Peptonization (hydrolysis)
  • pH Ind./Reagents: N/A
  • End Products: Amino Acids (crystalloid) / Amino Acids + Rennet Curd
  • (+) appearance: P: Juice / PC: Juice + Flakes
  • (-) appearance: Purple milk like control, blueberry shake, pink milk
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7
Q

Reduction

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Medium: Litmus Milk
  • Substrate: Litmus
  • Enzyme/Pathway: Reduction (Anaerobic Respiration)
  • pH Ind./Reagents: N/A
  • End Products: Leucolitmus (reduced litmus + ATP)
  • (+) appearance: R: bleached (bottom)
  • (-) appearance: Litmus colors (purple, pink, blue)
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8
Q

Reduction of Nitrates

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Medium: Nitrate Broth
  • Substrate: Nitrate
  • Enzyme/Pathway: Reduction of Nitrate / Anaerobic Respiration
  • pH Ind./Reagents: Pour out 1 mL, 3 drops of sulfanillic acid, 2 drops of dimethyl alpha naphthylamine solution
  • End Products: ATP + Nitrite
  • (+) appearance: Orange/Cherry Red
  • (-) appearance: Baby pink, beige
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9
Q

Production of Decarboxylase

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Medium: aa broth (1 aa, brom cresol purple, oil, basal medium)
  • Substrate: aa (ornithine, arg)
  • Enzyme/Pathway: decarboxylation/decarboxylase
  • pH Ind./Reagents: N/A
  • End Products: basic amine + CO2 gas
  • (+) appearance: Purple, Violet, Silvery
  • (-) appearance: Burgundy, Yellow
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10
Q

Hydrolysis of Urea

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Medium: Urea Broth (Phenol Red)
  • Substrate: Urea
  • Enzyme/Pathway: Hydrolysis of Urease
  • pH Ind./Reagents: N/A
  • End Products: Basic ammonia + CO2 gas
  • (+) appearance: Magenta
  • (-) appearance: Red, Yellow
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11
Q

Oxygen Requirements

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Obligate aerobe
  • Obligate Anaerobe
  • Facultative Anaerobe
  • Microaerophiles
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12
Q

Production of Hydrogen Sulfate

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Medium: SIM tall (contains Fe2+)
  • Substrate: Cysteine
  • Enzyme/Pathway: Production of H2S/ Anaerobic Respiration
  • pH Ind./Reagents: N/A
  • End Products: ATP + H2S gas
  • (+) appearance: Black Colloid (FeS)
  • (-) appearance: Like Control
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13
Q

Production of Indole

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Medium: SIM tall (containes Fe2+)
  • Substrate: Tryptophan
  • Enzyme/Pathway: Production of Indole
  • pH Ind./Reagents: Add 10 drops of Kovac’s Solution
  • End Products: Indole
  • (+) appearance: Red Kovac’s at top
  • (-) appearance: Yellow Kovac’s
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14
Q

Motility

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Medium: Sim or Motility Medium
  • Substrate: N/A
  • Enzyme/Pathway: N/A
  • pH Ind./Reagents: N/A
  • End Products: N/A
  • (+) appearance: feathery spread across stab line
  • (-) appearance: straight spread across stab line or close to stab line
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15
Q

Methyl Red Test

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Medium: MRVP Broth
  • Substrate: Glucose
  • Enzyme/Pathway: Fermentation
  • pH Ind./Reagents: Shake, Add 1mL to Clean test tube, 10 drops Methyl Red
  • End Products: ATP + Organic Strong Acids (4.4 or less)
  • (+) appearance: Red
  • (-) appearance: Yellow
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16
Q

Voges-Proskauer Test

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Medium: MRVP Broth
  • Substrate: glucose
  • Enzyme/Pathway: fermentation
  • pH Ind./Reagents: Shake, add 1mL to clean test tube, 10 drops alpha naphthol, 10 drops potassium hydroxase, creatine, stand for 20-25 minutes
  • End Products: ATP + AMC (acetyl methyl carbinol) + organic acids
  • (+) appearance: Ring of red at top of test tube
  • (-) appearance: no ring/yellow test tube
17
Q

Utilization of Unusual Nitrogen

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Medium: Ammonium Phosphate broth (brom cresol purple)
  • Substrate: Ammonium Phosphate
  • Enzyme/Pathway: Use of unusual Nitrogen/ammonium phosphate test
  • pH Ind./Reagents: N/A
  • End Products: Phosphoric Acid + Nitrogen is used
  • (+) appearance: Dirty Yellow
  • (-) appearance: Burgundy, Violet
18
Q

Utilization of Unusual Carbon

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Medium: Na Citrate Broth
  • Substrate: Citrate
  • Enzyme/Pathway: Use of unusual carbon/Na citrate test/Citratase
  • pH Ind./Reagents: N/A
  • End Products: Growth
  • (+) appearance: Cloudy, “shiggle” tube and see flakes like snowglobe, thick layer at top
  • (-) appearance: teeny bit at the bottom of the test tube (not bowl worth)
19
Q

Detection of Catalase

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Medium: Slant (any except blood)
  • Substrate: H2O2 (hydrogen peroxide)
  • Enzyme/Pathway: Catalase
  • pH Ind./Reagents: H2O2 FLOOD!
  • End Products: H2O + O2 gas
  • (+) appearance: bubbles
  • (-) appearance: no rxn
20
Q

MSA/Staph

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Medium:
  • Substrate:
  • Enzyme/Pathway:
  • pH Ind./Reagents: N/A
  • End Products:
  • (+) appearance:
  • (-) appearance:
21
Q

Coagulase Production

A
  • Medium:
  • Substrate:
  • Enzyme/Pathway:
  • pH Ind./Reagents: N/A
  • End Products:
  • (+) appearance:
  • (-) appearance:
22
Q

Latex Agglutination

Medium, Substrate, Enzyme/Pathway, pH Ind./Reagents, End prod., +/- app.

A
  • Medium:
  • Substrate:
  • Enzyme/Pathway:
  • pH Ind./Reagents: N/A
  • End Products:
  • (+) appearance:
  • (-) appearance:
23
Q

Hemolysin Production

A
  • α: partial lysis of hemoglobin, leaving greenish/brown discoloration around colonies
  • β: Total lysis of hemoglobin and RBCs, leaving clear colorles zones surrounding colonies
  • γ: Plasma proteins are used for growth, red (no change in medium)

microbio (17 decks)

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