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Liz's Microbiology Module 1 > Biochemicals > Flashcards

Biochemicals Flashcards

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1
Q

What are the end products of CHOs?

A

Acids

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2
Q

What makes alkaline end products?

A

Peptones

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3
Q

What order will organisms utilize CHOs and peptones?

A

CHOs 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

Methyl Red test

- What is in the media?

A

Glucose

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7
Q

Methyl Red test

- Reagent

A

Methyl red indicator (single drop at a time)

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8
Q

Methl Red test

- Results

A

Pos: red
Neg: yellow

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9
Q

Methyl Red test

  • Substrate(s)
  • End product
A

Substrate: glucose

End product: acid

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10
Q

Principle: to detect presence of the neutral end product acetone (acetylmethylcarbinol) resulting from the fermentation of glucose

A

Voges-Proskauer (VP) test

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11
Q

VP test

- Results

A

Pos: pink (butylene glycol fermentation)
Neg: no change (straw colored)

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12
Q

VP test

  • Substrate(s)
  • End product
A

Substrate: glucose

End product: acetoin (acetylmethylcarbinol)

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13
Q

VP test

- Reagent

A

Alpha napthol and KOH

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14
Q

VP test

- What is in the media?

A

Glucose

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15
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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16
Q

Citrate test

- What is in the media?

A

Citrate (inoculate slant)

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17
Q

Citrate test

  • Substrate(s)
  • End products
A

Substrate: citrate

End product: carbon?

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18
Q

Citrate test

- Reagent

A

None

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19
Q

Citrate test

- Results

A

Pos: blue
Neg: no change (green)

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20
Q

Principle: to detect an organism’s ability to oxidatively deaminate phenylalanine to phenylpyruvic acid

A

Phenylalanine deaminase

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21
Q

Phenylalanine deaminase

- What is in the media?

A

DL-phenylalanine agar (inoculate slant)

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22
Q

Phenylalanine deaminase

- Reagent

A

FeCl3 (ferric chloride)

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23
Q

Phenylalanine deaminase

  • Substrate(s)
  • End products
A

Substrate: deaminate phenylalanine

End product: phenylpyruvic acid

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24
Q

Another name for phenylalanine deaminase test

A

TDA test in some methods (API)

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25
Q

Phenylalanine deaminase

- Results

A

Pos: dark green slant (fades after several minutes)
Neg: no change (yellow from reagent)

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26
Q

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

A

Indole test

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27
Q

Indole test

- What is in the media?

A

Peptone or tryptone broth (little or no glucose)

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28
Q

Indole test

  • Substrate(s)
  • End product
A

Substrate: amino acid tryptophane

End product: indole

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29
Q

Indole test

  • Reagent (Ehrlich’s)
  • Reagent (Kovac’s)
A

SAME IN BOTH:

P-dimethylaminobenzaldehyde-HCl in ethyl alcohol

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30
Q

Indole test

- Results

A

Pos: red (middle layer in Ehrlich’s test, top layer in Kovac’s)
Neg: no change (yellow)

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31
Q

Two procedures for indole test

A

Ehrlich’s and Kovac’s

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32
Q

More sensitive test used for non-fermenting GNR’s and anaerobes where indole is extracted w/ chloroform, ether, or xylene

A

Ehrlich’s procedure (indole test)

33
Q

Procedure used for Enterobacteriaceae that is less sensitive

A

Kovac’s procedure (indole test)

34
Q

Principle: to identify an organism’s ability to ferment lactose and/or glucose, w/ or w/o the production of gas
- Production of H2S is also determined

A

Kligler iron agar

35
Q

Kliger iron test results:

  1. K/A
  2. A/A
  3. K/K
  4. CO2 gas production
  5. H2S production
A
  1. Glucose only (pink over yellow)
  2. Glucose + lactose + (yellow)
  3. No K/no change (red)
  4. Bubbles/cracks in media
  5. Black ppt
36
Q

Kliger iron

  • Substrate(s)
  • End product
A

Substrate: lactose and/or glucose and sodium thiosulfate

End products: glucose, galactose, and H2S

37
Q

Why should you not read Kliger iron results in the middle of the night?

A

All of the tubes will turn yellow (using up CHOs first) overnight, then use up the peptones (producing pink alkaline)

38
Q

Priniciple: to determine if an organism is motile (flagellated) or non-motile (non-flagellated)

A

Motility

39
Q

Sometimes used in motility test, these are colorless and reduced to formazan (red) when incorporated into cells. However, they’re not always used b/c they’re inhibitory to some organisms

A

Tetrazolium salts

40
Q

Motility test

- Results (w/o tetrazolium salts)

A

Pos: turbid
Neg: stab mark is obvious

41
Q

Principle: to determine an organism’s ability to split urea by the enzyme urease

A

Urease test

42
Q

Urease test

- Results

A

Pos: pH indicator turns pink

43
Q

Urease test

  • Substrate(s)
  • End products
A

Substrate: urea

End product: ammonia

44
Q

Urease test

- What is in the media?

A

Christensen’s (↓ buffer, urea, glucose, peptone, phenol red)

45
Q

Urease test

- Reagent

A

None

46
Q

Principle: to detect the presence of decarboxylase enzymes produced by organisms used in the breakdown of amino acids resulting in their corresponding amine + CO2

A

Decarboxylase media

47
Q

Decarboxylase media

  • Substrate(s)
  • End products
A

Substrate: lysine or ornithine

End product: alkaline products (cadaverine and putrescine)

48
Q

Decarboxylase media reactions:
Lysine → ?
Ornithine → ?

A

Lysine → cadaverine + CO2

Ornithine → putrecine + CO2

49
Q

Decarboxylase media

- Results

A

Pos: amino acids decarboxylated → alkaline pH (changes back to original purple/brown color)
Neg: glucose fermented first (acids turn yellow)

50
Q

Decarboxylase media

- What is in the media?

A

1% L-amino acid, bromcresol purple and cresol red (indicator), glucose

51
Q

Decarboxylase media

- Reagent

A

None

52
Q

Principle: to detect the presence of arginine dihydrolase enzymes caused by some bacteria in the breakdown of the amino acid, argentine

A

Arginine Dihydrolase

53
Q

Arginine dihydrolase

- What is in the media?

A

Moeller’s decarboxylase media

54
Q

Arginine dihydrolase

  • Substrate(s)
  • End products
A

Substrate: arginine

End product: arginine dihydrolase

55
Q

Arginine dihydrolase

- Results

A

Same as decarboxylase test:
Pos: amino acids decarboxylated → alkaline pH (changes back to original purple/brown color)
Neg: glucose fermented first (acids turn yellow)

56
Q

What is more sensitive for H2S detection than KIA?

A

Lead acetate test

57
Q

Lead actetate

  • Indicator
  • End product
A

Indicator: lead acetate

End product: H2S

58
Q

Lead acetate test

- Results

A

Filter paper w/ lead acetate turns black

59
Q

Principle: to detect an organism’s ability to degrade specific CHOs w/ the production of acid, w/ or w/o visible gas

A

1% CHOs

60
Q

1% CHOs

- What is in the media?

A
  • CHO-free base (pruple broth or pheno red base)
  • 1% or 0.5% CHOs (monosaccharides, polysaccharide, or polyhydric alcohol)
  • Contains Durham tube (smaller inverted tube w/in larger tube
61
Q

1% CHOs

  • Substrate(s)
  • End product
A

Substrate: various CHOs (1/tube)

End product: ???

62
Q

1% CHOs

- Reagent

A

None

63
Q

1% CHOs

- Results

A

Depends on which base is used…
When purple broth base…
Pos: yellow (acid)
Neg: purple

64
Q

Principle: to determine an organism’s mechanism for utilization of glucose; fermentative, oxidative, or neither

A

Oxidation/Fermentation (OF)

65
Q

Oxidation/Fermentation (OF)

- What is in the media for Hugh and Leifson’s 2 tube test?

A
  • High concentration of CHO

- Low concentration of peptone

66
Q

OF

  • Substrate(s)
  • End product
A

Substrate: glucose (or other CHOs)

End product: acid end product

67
Q

OF

- Reagent

A

None

68
Q

OF

- Results

A

Fermenter: acid (yellow) in oiled tube
Oxidizer: acid (yellow) in unoiled tube only
Non-oxidizer (asaccharolytic): acid (yellow) in neither tube (stays green)

69
Q

Principle: to determine the presence of oxidized cytochrome C (cytochrome oxidase enzyme)

A

Cytochrome oxidase (oxidase test)

70
Q

Cytochrome oxidase test

- Reagents

A
  • 1% tetramethyl-p-phenylenediamine dihydrochloride

- Dimethyl-p-phenylendiamine + alpha napthol

71
Q

Cytochrome oxidase test

- 3 testing methods

A
  • Filter paper
  • Plate or tube
  • Impregnated packaged strips
72
Q

Cytochrome oxidase test

- Results

A 
Filter paper method
- Pos: blue mark
- Neg: no change
Plate method
- Pos: growth
- Neg: no growth (kills organism)
73
Q

Principle: to determine an organism’s ability to reduce nitrate to nitrite - or nitrate to nitrogen gas

A

Nitrate test

74
Q

Nitrate test

- What is in the media?

A

Potassium nitrate

75
Q

Nitrate test

  • Substrate(s)
  • End product
A

Substrate: NO3

End product: Nitrite or nitrogen gas

76
Q

Nitrate test

- Reagents

A
  • Alpha napththylamine or dimethyl-apha-napthylamine
  • Sufonilic aid
  • Zinc Dust (control)
77
Q

Nitrate test results

  • NO3 → NO2
  • NO3 → N2
  • NO3 → not reduced
A
  • Red after addition of two reagents (pos)
  • Clear after addition of two reagents AND clear after add-on of zinc dust (pos)
  • clear after addition of two reagents AND red addition of zinc dust (neg)
78
Q

Principle: Testing organism for production of beta-glucuronidase

A

MUG test

79
Q

MUG test

- Results

A

Pos: fluorescence w/ UV light

- Helpful screening for E. coli

Liz's Microbiology Module 1 (11 decks)

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