Biochemicals Flashcards
(79 cards)
1
Q
What are the end products of CHOs?
A
Acids
2
Q
What makes alkaline end products?
A
Peptones
3
Q
What order will organisms utilize CHOs and peptones?
A
CHOs first. peptones next
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
5
Q
Two types of fermentation
A
- Mixed acid fermentation
- Butylene glycol fermentation
6
Q
Methyl Red test
- What is in the media?
A
Glucose
7
Q
Methyl Red test
- Reagent
A
Methyl red indicator (single drop at a time)
8
Q
Methl Red test
- Results
A
Pos: red
Neg: yellow
9
Q
Methyl Red test
- Substrate(s)
- End product
A
Substrate: glucose
End product: acid
10
Q
Principle: to detect presence of the neutral end product acetone (acetylmethylcarbinol) resulting from the fermentation of glucose
A
Voges-Proskauer (VP) test
11
Q
VP test
- Results
A
Pos: pink (butylene glycol fermentation)
Neg: no change (straw colored)
12
Q
VP test
- Substrate(s)
- End product
A
Substrate: glucose
End product: acetoin (acetylmethylcarbinol)
13
Q
VP test
- Reagent
A
Alpha napthol and KOH
14
Q
VP test
- What is in the media?
A
Glucose
15
Q
Principle: to determine if an organism is capable of utilizing citrate as its sole source of carbon
A
Sodium Citrate (Citrate) test
16
Q
Citrate test
- What is in the media?
A
Citrate (inoculate slant)
17
Q
Citrate test
- Substrate(s)
- End products
A
Substrate: citrate
End product: carbon?
18
Q
Citrate test
- Reagent
A
None
19
Q
Citrate test
- Results
A
Pos: blue
Neg: no change (green)
20
Q
Principle: to detect an organism’s ability to oxidatively deaminate phenylalanine to phenylpyruvic acid
A
Phenylalanine deaminase
21
Q
Phenylalanine deaminase
- What is in the media?
A
DL-phenylalanine agar (inoculate slant)
22
Q
Phenylalanine deaminase
- Reagent
A
FeCl3 (ferric chloride)
23
Q
Phenylalanine deaminase
- Substrate(s)
- End products
A
Substrate: deaminate phenylalanine
End product: phenylpyruvic acid
24
Q
Another name for phenylalanine deaminase test
A
TDA test in some methods (API)
25
Phenylalanine deaminase
| - Results
Pos: dark green slant (fades after several minutes)
Neg: no change (yellow from reagent)
26
Principle: to detect whether an organism can split indole from the amino acid tryptophane
Indole test
27
Indole test
| - What is in the media?
Peptone or tryptone broth (little or no glucose)
28
Indole test
- Substrate(s)
- End product
Substrate: amino acid tryptophane
| End product: indole
29
Indole test
- Reagent (Ehrlich's)
- Reagent (Kovac's)
SAME IN BOTH:
| P-dimethylaminobenzaldehyde-HCl in ethyl alcohol
30
Indole test
| - Results
Pos: red (middle layer in Ehrlich's test, top layer in Kovac's)
Neg: no change (yellow)
31
Two procedures for indole test
Ehrlich's and Kovac's
32
More sensitive test used for non-fermenting GNR's and anaerobes where indole is extracted w/ chloroform, ether, or xylene
Ehrlich's procedure (indole test)
33
Procedure used for Enterobacteriaceae that is less sensitive
Kovac's procedure (indole test)
34
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
Kligler iron agar
35
Kliger 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/cracks in media
5. Black ppt
36
Kliger iron
- Substrate(s)
- End product
Substrate: lactose and/or glucose and sodium thiosulfate
| End products: glucose, galactose, and H2S
37
Why should you not read Kliger iron results in the middle of the night?
All of the tubes will turn yellow (using up CHOs first) overnight, then use up the peptones (producing pink alkaline)
38
Priniciple: to determine if an organism is motile (flagellated) or non-motile (non-flagellated)
Motility
39
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
Tetrazolium salts
40
Motility test
| - Results (w/o tetrazolium salts)
Pos: turbid
Neg: stab mark is obvious
41
Principle: to determine an organism's ability to split urea by the enzyme urease
Urease test
42
Urease test
| - Results
Pos: pH indicator turns pink
43
Urease test
- Substrate(s)
- End products
Substrate: urea
| End product: ammonia
44
Urease test
| - What is in the media?
Christensen's (↓ buffer, urea, glucose, peptone, phenol red)
45
Urease test
| - Reagent
None
46
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
47
Decarboxylase media
- Substrate(s)
- End products
Substrate: lysine or ornithine
| End product: alkaline products (cadaverine and putrescine)
48
Decarboxylase media reactions:
Lysine → ?
Ornithine → ?
Lysine → cadaverine + CO2
| Ornithine → putrecine + CO2
49
Decarboxylase media
| - Results
Pos: amino acids decarboxylated → alkaline pH (changes back to original purple/brown color)
Neg: glucose fermented first (acids turn yellow)
50
Decarboxylase media
| - What is in the media?
1% L-amino acid, bromcresol purple and cresol red (indicator), glucose
51
Decarboxylase media
| - Reagent
None
52
Principle: to detect the presence of arginine dihydrolase enzymes caused by some bacteria in the breakdown of the amino acid, argentine
Arginine Dihydrolase
53
Arginine dihydrolase
| - What is in the media?
Moeller's decarboxylase media
54
Arginine dihydrolase
- Substrate(s)
- End products
Substrate: arginine
| End product: arginine dihydrolase
55
Arginine dihydrolase
| - Results
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
What is more sensitive for H2S detection than KIA?
Lead acetate test
57
Lead actetate
- Indicator
- End product
Indicator: lead acetate
| End product: H2S
58
Lead acetate test
| - Results
Filter paper w/ lead acetate turns black
59
Principle: to detect an organism's ability to degrade specific CHOs w/ the production of acid, w/ or w/o visible gas
1% CHOs
60
1% CHOs
| - What is in the media?
- 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
1% CHOs
- Substrate(s)
- End product
Substrate: various CHOs (1/tube)
| End product: ???
62
1% CHOs
| - Reagent
None
63
1% CHOs
| - Results
Depends on which base is used...
When purple broth base...
Pos: yellow (acid)
Neg: purple
64
Principle: to determine an organism's mechanism for utilization of glucose; fermentative, oxidative, or neither
Oxidation/Fermentation (OF)
65
Oxidation/Fermentation (OF)
| - What is in the media for Hugh and Leifson's 2 tube test?
- High concentration of CHO
| - Low concentration of peptone
66
OF
- Substrate(s)
- End product
Substrate: glucose (or other CHOs)
| End product: acid end product
67
OF
| - Reagent
None
68
OF
| - Results
Fermenter: acid (yellow) in oiled tube
Oxidizer: acid (yellow) in unoiled tube only
Non-oxidizer (asaccharolytic): acid (yellow) in neither tube (stays green)
69
Principle: to determine the presence of oxidized cytochrome C (cytochrome oxidase enzyme)
Cytochrome oxidase (oxidase test)
70
Cytochrome oxidase test
| - Reagents
- 1% tetramethyl-p-phenylenediamine dihydrochloride
| - Dimethyl-p-phenylendiamine + alpha napthol
71
Cytochrome oxidase test
| - 3 testing methods
- Filter paper
- Plate or tube
- Impregnated packaged strips
72
Cytochrome oxidase test
| - Results
```
Filter paper method
- Pos: blue mark
- Neg: no change
Plate method
- Pos: growth
- Neg: no growth (kills organism)
```
73
Principle: to determine an organism's ability to reduce nitrate to nitrite - or nitrate to nitrogen gas
Nitrate test
74
Nitrate test
| - What is in the media?
Potassium nitrate
75
Nitrate test
- Substrate(s)
- End product
Substrate: NO3
| End product: Nitrite or nitrogen gas
76
Nitrate test
| - Reagents
- Alpha napththylamine or dimethyl-apha-napthylamine
- Sufonilic aid
- Zinc Dust (control)
77
Nitrate test results
- NO3 → NO2
- NO3 → N2
- NO3 → not reduced
- 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
Principle: Testing organism for production of beta-glucuronidase
MUG test
79
MUG test
| - Results
Pos: fluorescence w/ UV light
| - Helpful screening for E. coli
