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Biochemicals Flashcards

1
Q

What are the end products of carbohydrates?

A

acids

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2
Q

What make alkaline end products?

A

Peptones

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3
Q

What order will organism utilize carbohydrates and peptones?

A

They will use CHO first, peptones next

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4
Q

Principle: to determine the ability of an organism to produce large amounts of acid or neutral end products from glucose fermentation

A

Methyl Red Test

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5
Q

Two types of fermentation

A
    • Mixed Acid Fermentation

- Butylene Glycol fermentation

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6
Q

What is used in Methyl Red Test’s media?

A

Glucose

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7
Q

What is the reagent used in methyl red test?

A

Methyl Red Indicator (single drop at a time)

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8
Q

Methyl Red Test results

A

Red - positive

Yellow- negative

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9
Q

Methyl Red Test

  • Substrate(s)
  • Endproduct?
  • Reagent?
A

Substrate: glucose
End products: acid
Reagent: methyl red indicator

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10
Q

Principle: to detect presence of the neutral endproduct acetone (acetylmethycarbinol) resulting from the fermentation of glucose

A

Voges-Proskauer Test

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11
Q

VP Test results

A 
Pink - positive 
No change (straw colored) - negative
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12
Q

VP Test

  • Substrate(s)
  • Endproduct?
A

Substrate: glucose
Endproduct: acetoin (acetylmethylcarbinol)

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13
Q

VP Test

- Reagent

A

Reagent: Alpha napthol and KOH

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14
Q

Principle: To determine if an organism is capable of utilizing citrate as its sole source of carbon

A

Sodium Citrate (Citrate) Test

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15
Q

Citrate Test Results

A

On a slant!
Blue - positive
No change (green) - negative

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16
Q

Sodium Citrate (Citrate) Test

  • Substrate
  • Endproduct
A

Substrate: citrate
Endproduct: carbon?

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17
Q

Principle: to detect an organisms ability to oxidatively deaminate phenylalanine to phenylpyruvic acid.

A

Phenylalanine Deaminase

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18
Q

Another name for Phenylalanine Deaminase Test?

A

TDA test in some methods (API)

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19
Q

Phenylalanine Deaminase Test Results

A

Dark green slant - positive test (fades after several minutes)

No change (yellow from reagent) - negative test

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20
Q

Phenylalanine Deaminase

  • Substrate(s)
  • Endproduct
A

Substrate: deaminate phenylalanine
Endproduct: phenylpyruvic acid

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21
Q

Phenylalanine Deaminase

- Reagent

A

FeCl3 (Ferric Chloride)

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22
Q

Principle: to detect whether an organism can split indole from the amino acid tryptophane

A

Indole Test

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23
Q

Indole Test Results

A 
Red - positive (middle layer in Ehrlich's Test, top layer in Kovac's) 
No change (yellow) - negative
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24
Q

Indole Test

  • Substrates
  • Endproduct
A

Substrates: amino acid tryptophane
Endproduct: indole

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25
Indole Test - Reagent (Ehrlich's) - Reagent (Kovac's)
SAME IN BOTH: | P-dimethylaminobenzaldehyde-HCLin ethyl alcohol
26
Two procedures for Indole Test
Ehrlich's Procedure | Kovac's Procedure
27
Indole test media
Peptone or tryptone broth (Little or no glucose!)
28
More sensitive test used for non fermenting GNR's and anaerobes where indole is extracted with chloroform, ether, or xylene
Ehrlich's Procedure (Indole Test)
29
Procedure used for Enterobacteriaceae that is less sensitive
Kovac's Procedure (Indole Test)
30
Principle: To identify an organism's ability to ferment lactose and / or glucose, with or without the production of gas.. The production of hydrogen sulfide is also determined
Kligler Iron Agar
31
Kligler Iron Test Results 1. K/A 2. A/A 3. K/K 4. CO2 gas production 5. H2S production
1. glucose only (pink over yellow) 2. glucose + lactose + (Yellow) 3. no K/ no change (red) 4. bubbles 5. black ppt
32
Kligler Iron - Substrate(s) - Endproduct
Substrates: lactose and or glucose and sodium thiosulfate | *Endproducts: glucose, galactose, and H2S
33
Why should you not read Kligler Iron results in the middle of the night?
All of the tubes will turn yellow (using up carbohydrates first) over night then use up the peptones (producing pink alkaline)
34
Principle: to determine if an organism is motile (flagellated) or non motile (non flagellated)
Motility
35
Sometimes used in Motility testing, these are colorless and reduced to formazan (red) when incorporated into cells. However, they are not always used because are inhibitory to some organisms
Tetrazolium Salts
36
Motility Test Results without tetrazolium salts
turbid- positive | stab mark obvious - negative
37
Principle: determine an organisms ability to split urea by the enzyme urease
Urease Test
38
Urease Test results
pH indicator turns Pink - positive
39
Urease Test - Substrates - Endproduct
substance: urea endproduct: ammonia
40
Principle: to detect the presence of decarboxylase enzymes produced by organisms used in the breakdown of amino acids resulting in their corresponding amine + CO2
Decarboxylase Media
41
Decarboxylase Media - substrates - endproducts
sub: amino acids end: alkaline endproducts (cadaverine and putrescine)
42
Decarboxylase Media reactions Lysine---------> ? Ornithine----------> ?
cadaverine + CO2 putrecine + CO2
43
Decarboxylase Media Test results
Glucose fermented first - acids turn yellow (neg) Amino acids decarboxylated - alkaline pH ( changes back to original purple/brown color) - positive test
44
Principle: to detect the presence of arginine dihydrolase enzymes used by some bacteria in the breakdown of the amino acid, argentine
Arginine Dihydrolase
45
Arginine Dihydrolase - sub - end
sub: arginine end: arginine dihydrolase
46
Arginine Dihydrolase Test results
Same as decarboxylase tests: Glucose fermented first - acids turn yellow (neg) Amino acids decarboxylated - alkaline pH ( changes back to original purple/brown color) - positive test
47
What test is more sensitive for H2S detection than KIA?
Lead Acetate Test
48
Lead Acetate Test: - indicator - Endproducts
Indicator: lead acetate End: H2S
49
Lead Acetate Test | -Test results
-filter paper with lead acetate turns black
50
Principle: to detect an organism's ability to degrade specific carbohydrates with the production of acid, with or without visible gas
1% Carbohydrates
51
1% Carbohydrates | - test results
- When purple broth base yellow (acid) = positive purple = negative
52
1% Carbs - sub - end?
???
53
Principle: to determine an organism's mechanism for utilization of glucose; fermentative, oxidative or neither
Oxidation/Fermentation (OF)
54
Oxidation/Fermentation (OF) | - Media: Hough and Leifson's two tube test
- High concentration of carbohydrate | - Low concentration of peptone
55
Oxidation/Fermentation (OF) - Sub - End
- sub: glucose | - end: acid end products
56
Oxidation/Fermentation (OF) | - Test results
- acid (yellow) in oiled tube = fermenter - acid (yellow) in uncoiled tube only = oxidizer - acid (yellow) in neither tube (stays green) = asaccharolytic ( non-oxidizer)
57
Principle: To determine the presence of oxidized cytochrome C (cytochrome oxidase enzyme)
Cytochrome Oxidase (Oxidase Test)
58
``` Cytochrome Oxidase (Oxidase Test) - reagents ```
- 1% tetramethyl-p-phenylenediamine dihydrochloride | - Dimethyl-p-phenylendiamine + alpha napthol
59
``` Cytochrome Oxidase (Oxidase Test) - 3 testing methods ```
- Filter paper - plate or Tube - impregnated packaged strips
60
``` Cytochrome Oxidase (Oxidase Test) - test results ```
- Filter paper = blue mark | - Plate = kills organism
61
Principle: To determine an organism's ability to reduce nitrate to nitrite - or nitrate to nitrogen gas
Nitrate Test
62
Nitrate Test - Sub: - End:
- Sub: nitrate NO3 | - End: nitrite or nitrogen gas
63
Nitrate Test | - reagents
- Alpha napththylamine or dimethyl-apha-napthylamine - Sufonilic aid - Zinc Dust (control)
64
Nitrate Test Results - NO3 -------> NO2 - NO3 -------> N2 - NO3 -------> not reduced
- red after addition of two regents (pos test - clear after addition of tow reagents AND clear after add-on of zinc dust (pos test) - clear after addition of two reagents AND red after addition of Zinc dust (neg test)
65
Testing organism for production of Beta- glucuronidase
MUG Test
66
MUG Test | - test results
- fluorescence with UV light | - helpful screening for E. coli

Microbiology (13 decks)

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