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Microbiology 210 > Final Prep Lab Exam 2 > Flashcards

Final Prep Lab Exam 2 Flashcards

Lab Quiz 3, Lab Quiz 4, Protozoan Table, SG, +

1
Q

What is the purpose of the decarboxylase test?

5-11: Decarboxylase

A

To determine whether or not bacteria can decarboxylate lysine or ornithine

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2
Q

What is the purpose of the deaminase test?

5-12: Deaminase

A

To determine whether bacteria can deaminate phenylalanine

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3
Q

What is the purpose of the casease test?

5-16: Casease

A

To determine whether bacteria can hydrolyze casein

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4
Q

What is the purpose of the urease test?

5-18 Urease

A

To determine if bacteria can hydrolyze urea

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5
Q

What is the purpose of the TSI agar test?

5-21: TSI Agar H2S test

A

To differentiate bacteria on the basis
of glucose fermentation, lactose fermentation, sucrose fermentation, and sulfur reduction

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6
Q

What is the purpose of the oxidase test?

5-7: Oxidase

A

To test bacteria for oxidase enzyme

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7
Q

What is the purpose of the catalase test?

5-6: Catalase

A

To test bacteria for the catalase enzyme

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8
Q

What is the purpose of the anaerobic jar test?

2-8: Anaerobic Jar

A

To determine the oxygen requirements of an organism

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9
Q

What is the name of the medium?

5-11: Decarboxylase

A

Lysine/ornothine broth
- LDC or ODC

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10
Q

What is the substrate, bacterial enzyme, and end product?

5-11: Decarboxylase

A

Substrate: Lysine or Ornothine
Bacterial Enzyme: Lysine or Ornothine Decarboxylase (endoenzyme)
End Products: Cataverine or putrescine (alkaline diamines) + CO2 g

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11
Q

What is the appearance of a positive/negative?

5-11: Decarboxylase

A

Positive: Purple/gray
Negative: Yellow

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12
Q

What is the reagent and pH indicator?

5-11: Decarboxylase

A
  • None
  • Bromocresol purple
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13
Q

What else is important about this test? (4)

5-11: Decarboxylase

A
  • Results cannot be read before 48H due to pH bounce
  • Anaerobic enzymes (oil)
  • Decarboxylation is anaerobic (oil used)
  • Glucose is used first then acids which activate decarboxylase enzymes
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14
Q

What is the name of the medium?

5-12: Deaminase

A

Phenylalanine agar

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15
Q

What is the substrate, bacterial enzyme, and end product?

5-12: Deaminase

A

Substrate: Phenylalanine
Bacterial enzyme: Phenylalanine deaminase (endoenzyme)
End product: Ammonia + phenylpyruvic acid

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16
Q

What is the appearance of a positive/negative?

5-12: Deaminase

A

Positive: Green
Negative: Yellow

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17
Q

What is the reagent and pH indicator?

5-12: Deaminase

A

Reagent: Ferric Chloride (FeCl3) (indicates phenols)
pH indicator: None

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18
Q

What else is important about this test?(2)

5-12: Deaminase

A
  • pH products neutralize eachother
  • Aerobic enzyme
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19
Q

What is the name of the medium?

5-16: Casease

A

Milk agar

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20
Q

What is the substrate, bacterial enzyme and end products?

5-16: Casease

A

Substrate: Casein
Bacterial enzyme: Casease (exoenzyme)
End products: AA + Polypeptides

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21
Q

What is a positive/negative result?

5-16: Casease

A

Positive: Clear zone
Negative: White

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22
Q

Are there reagents or pH indicators?

5-16: Casease

A

None

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23
Q

What is the name of the medium?

2-8: Anaerobic Jar

A

Nutrient agar

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24
Q

What else is important about this test?(1)

5-16: Casease

A
  • Both + and - organisms still grow
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25
Q

What is the name of the medium?

5-18 Urease

A

Urea tablet

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26
Q

What is the substrate, bacterial enzyme, and end products?

5-18 Urease

A

Substrate: Urea
Bacterial enzyme: Urease
End products: Ammonia + CO2 g

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27
Q

What is a positive/negative result?

5-18 Urease

A

Positive: Pink
Negative: Yellow

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28
Q

What are the reagents an pH indicators?

5-18 Urease

A

None
Phenol Red

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29
Q

What else is important about this test?(2)

5-18 Urease

A
  • Ammonia burns mucous membranes and neutralizes stomach acid
  • H. pylori
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30
Q

What is the name of the medium?

5-21: TSI Agar H2S test

A

TSI agar

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31
Q

What are the substrates, bacterial enzymes, and end products?

5-21: TSI Agar H2S test

A

Substrates: Cystine + Sodium thiosulfate
Bacterial enzyme: Cystine desulfurase + Thiosulfate reductase (endoenzymes)
End product: H2S

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32
Q

What is a positive/negative result?

5-21: TSI Agar H2S test

A

Positive: Any black
Negative: No Black

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33
Q

What is the reagent and H2S indicator?

5-21: TSI Agar H2S test

A

No reagent
Ferrous ammonium sulfate

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34
Q

What else is important to consider?(1)

5-21: TSI Agar H2S test

A

Black color indicates an acidic pH

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35
Q

What is the name of the medium?

5-21: TSI Agar pH test

A

TSI agar

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36
Q

What are the substrates, bacterial enzymes, and end products?

5-21: TSI Agar pH test

A

Substrates: Glucose, Lactose, Sucrose
Bacterial enzymes: Various endoenzymes
End products: Acids

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37
Q

What does a positive/negative result look like?

5-21: TSI Agar pH test

A

Positive: Some yellow (glucose only)
Positive: All yellow (glucose plus at least one dissacharide)
Negative: No yellow or black

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38
Q

What are the reagents and pH indicator used?

5-21: TSI Agar pH test

A

None
Phenol Red

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39
Q

What else is important about this test?(1)

5-21: TSI Agar pH test

A
  • In glucose only ferm. (K/A slant) glucose was in low % and runs out, peptones are then used aerobically
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40
Q

What is the name of the medium?

5-7: Oxidase

A

Nutrient agar

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41
Q

What is the substrate, bacterial enzyme, and end product?

5-7: Oxidase

A

Substrate: O2
Bacterial enzyme: Cytochrome C oxidase (endoenzyme)
End product: H2O

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42
Q

What is a positive/negative result?

5-7: Oxidase

A

Positive: Red/pink
Negative: No change

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43
Q

What is the reagent and pH indicator?

5-7: Oxidase

A

Oxidase reagent
None

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44
Q

What is the name of the medium?

5-6: Catalase

A

Nutrient agar

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45
Q

What is the substrate, bacterial enzyme, and end products?

5-6: Catalase

A

Substrate: H2O2
Bacterial enzyme: Catalase (endoenzyme)
End products: H2O + O2 g

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46
Q

What is a positive/ negative result?

5-6: Catalase

A

Positive: Bubbling
Negative: No bubbles

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47
Q

What reagent and pH indicator were used?

5-6: Catalase

A

H2O2
None

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48
Q

What is the substrate, chemical, and end product?

2-8: Anaerobic Jar

A

Substrate: O2
Chemical: Anaerogen chemical
End product: H2O

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49
Q

What is a positive/negative result?

2-8: Anaerobic Jar

A

Positive: Growth
Negative: No growth

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50
Q

What is the purpose of the thiobroth test?

2-7: Thiobroth

A

To determine the O2 requirements of
an organism

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51
Q

What is the name of the medium?

2-7: Thiobroth

A

Thio broth

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52
Q

What is the purpose of the sodium thioglycollate and L-cystine in the recipe?

2-7: Thiobroth

A

Remove O2 to make anaerobic environment (turned to water)

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53
Q

What do these results mean?

2-7: Thiobroth

A
  • Growth only at top means the organism is a obligate aerobe
  • Growth at top and in stab means the organism is a facultative anaerobe
  • Growth only in the stab means obligate anaerobe
  • Thin growth near the top means organism is a microaerophile
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54
Q

What enzyme is present in aerobic bacteria? Anaerobic?

2-7: Thiobroth

A

Superoxide dismutase - aerobic
Catalase - anaerobic

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55
Q

What are the results of this test?

5-21: TSI Agar HS2 and pH

A
  • K/A H2S-
  • A/A H2S-
  • K/K H2S-
  • K/K H2S-
  • A/A H2S+
  • K/A H2S+
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56
Q

Is protein deamination and decarboxylation an anaerobic or aerobic process?

A

Deamination - aerobic
Decarboxylation - anaerobic

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57
Q

What is the purpose of a Gelatinase test?

5-17: Gelatinase

A

Allows us to determine the ability of a microbe to produce gelatinases

Ex. Clostridium perfringens (Gangrene)

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58
Q

What is the purpose of a motility test?

5-28: Motility Test

A

To be able to detect bacterial motility which is an important differential characteristic

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59
Q

What is the purpose of temperature on microbial growth?

2-9: Effect of Temperature on Microbial Growth

A

A qualitative procedure to understand the effect of temperature on bacterial growth

Listeria monocytogenes is a psychrotroph that lived in your fridge and contaminates dairy products

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60
Q

What is the purpose of a nitrate reduction test?

5-8: Nitrate Reduction Test, First Step

A

This test tells us if bacteria can use nitrate as a final electron acceptor in anaerobic respiration. This is also to test if the gram-negative rods either do not reduce nitrate or reduce it beyond nitrite.

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61
Q

What is the purpose of the Kirby-Bauer Test?

7-3: Kirby-Bauer Test

A

To perform a culture and antibiotic sensitivity test on an organism

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62
Q

What is the purpose of the Membrane Filtration Test?

8-12: Membrane Filtration Test

A

To test water samples for the presence of coliform bacteria (presumptive)

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63
Q

What is the name of the medium?

5-17: Gelatinase

A

Gelatin Strip

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64
Q

What is the substrate, enzyme, and product(s)?

5-17: Gelatinase

A

Substrate: Gelatin
Enzyme: Exoenzyme Gelatinase
Products: AA + Polypeptides

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65
Q

What is the appearance of a positive test? Negative?

5-17: Gelatinase

A

Positive: Blue
Negative: No Change

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66
Q

What were the reagents used? pH indicator?

5-17: Gelatinase

A

None

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67
Q

What were the two bacteria that was used?

5-17: Gelatinase

A

2 type of bacteria:
– Pseudomonas aeruginosa (Pa) +
– E. coli (Ec) -

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68
Q

What was the medium of this experiment?

5-28: Motility Test

A

TTC Agar

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69
Q

What is the substrate, enzyme, and end product(s)?

5-28: Motility Test

A

Substrate: TTC
Enzyme: Various Reductases
Product: Formazan

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70
Q

What is the appearance of a positive? Negative?

5-28: Motility Test

A

Positive: Fuzzy Red Growth
Negative: Solid Non-fuzzy Red Line

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71
Q

What reagents and pH indicator were used?

5-28: Motility Test

A

None

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72
Q

What is the medium?

5-8: Nitrate Reduction Test, First Step

A

Nitrate Broth

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73
Q

What is the substrate, enzyme, and end product(s)?

5-8: Nitrate Reduction Test, First Step

A

Substrate: Nitrate (NO3)
Enzyme: Endoenzyme nitrate reductase
Product: Nitrite (NO2)

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74
Q

What is the appearance of a positive? Negative?

5-8: Nitrate Reduction Test, First Step

A

Positive: Red
Negative: Proceed to step 2

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75
Q

What reagent and pH indicators were used in step 1?

5-8: Nitrate Reduction Test, First Step

A

Reagent: Nitrate reagent A + B
pH indicator: None

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76
Q

What is the substrate, enzyme, and end product(s)?

5-8: Nitrate Reduction Test, Second Step

A

Substrate: Nitrate (NO3)
Enzyme: Many endoenzyme nitrate reductases
Products: Ammonium (NH4+, no gas) or NO,N2O,N2 gases (denitrification)

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77
Q

What is the appearance of a positive result? Negative?

5-8: Nitrate Reduction Test, Second Step

A

Positive: Clear after zinc
Negative: Red after zinc

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78
Q

What reagent and pH indicator were used?

5-8: Nitrate Reduction Test, Second Step

A

Reagent: Nitrate reagents A, B, Zinc
pH indicator: None

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79
Q

What is important about the zinc added in step two?

5-8: Nitrate Reduction Test, Second Step

A

If the broth turns red it is because nitrate was reduced by zinc. If it does not it means there was no nitrate(NO3) to reduce because it had already been reduced beyond nitrite (NO2)

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80
Q

What was the medium of this test? (5)

7-3: Kirby-Bauer Test

A

Mueller-Hinton Agar standardizes the following
- pH:7.4
- Soft agar
- 4mm depth agar for lateral diffusion
- Temp: 37 C
- Amount of drug

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81
Q

What were the three levels of susceptibility?

7-3: Kirby-Bauer Test

A

Sensitive: A normal dose is effective against bacteria
Intermediate: A higher than normal dose of an antibiotic is required
Resistant: No acceptable dose is effective

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82
Q

What is synergy, broad spectrum, narrow spectrum?

7-3: Kirby-Bauer Test

A

Synergy: when the effect of two drugs combines to be greater than their sum
Broad spectrum: Drugs that are effective against most bacteria typically reserved as final options as it can harm native flora
Narrow spectrum: Highly targetted drugs that are effective against a specific mecanism of bacterial growth. Must know what bacteria you are dealing with

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83
Q

What was the medium of this experiment?

8-12: Membrane Filtration Test

A

Endo agar or Nutrient Agar

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84
Q

What is the appearance of a positive? Negative?

8-12: Membrane Filtration Test

A

Positive:
Green Metalic Sheen (rapid lactose fermenter)
Dark pink/Red (slow lactose fermenter)
Negative:
Light Pink/Colorless (no lactose fermentation)

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85
Q

Entamoeba histolytica; Disease, Motility, Stages, Infectious Stage(s) and mode of Transmission.

A
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86
Q

Balantidium coli; Disease, Motility, Stages, Infectious Stage(s) and mode of Transmission.

A
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87
Q

Giardia lamlia (intestinalis); Disease, Motility, Stages, Infectious Stage(s) and mode of Transmission.

A
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88
Q

Trichomonas vaginalis; Disease, Motility, Stages, Infectious Stage(s) and mode of Transmission.

A
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89
Q

Trypanosoma brucei; Disease Motility, Stages, Infectious Stage(s) and mode of Transmission.

A
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90
Q

Toxoplasma gondii; Disease Motility, Stages, Infectious Stage(s) and mode of Transmission.

A
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91
Q

Plasmodium falciparum; Disease Motility, Stages, Infectious Stage(s) and mode of Transmission.

A
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92
Q

Entamoeba histolytica; Symptoms, Treatment, Diagnosis/Notes

A
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93
Q

Balantidium coli; Symptoms, Treatment, Diagnosis/Notes

A
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94
Q

Giardia lamblia (intestinalis); Symptoms, Treatment, Diagnosis/Notes

A
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95
Q

Trichomonas vaginalis; Symptoms, Treatment, Diagnosis/Notes

A
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96
Q

Trypanosoma brucei; Symptoms, Treatment, Diagnosis/Notes

A
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97
Q

Toxoplasma gondii; Symptoms, Treatment, Diagnosis/Notes

A
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98
Q

Plasmodium falciparum; Symptoms, Treatment, Diagnosis/Notes

A
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99
Q

What is this?

A

Entamoeba histolytica (cyst)

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100
Q

What is this?

A

Entamoeba histolytica (trophozoite)

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101
Q

What is this?

A

Balantidium coli (cyst)

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102
Q

What is this?

A

Balantidium coli (Trophozoite)

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103
Q

What is this?

A

Giardia lamblia (intestinalis) (cyst)

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104
Q

What is this?

A

Giardia lamblia (intestinalis) (trophozoite)

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105
Q

What is this?

A

Trichomonas vaginalis (Trophozoite)

106
Q

What is this?

A

Trypanosoma brucei (Trypomastigote)

107
Q

What is this?

A

Toxoplasma gondii (Trophozoite)

108
Q

What is this?

A

Plasmodium falciparum (Ring stage, merozoites)

109
Q

Enzymes called decarboxylases catalyze the removal of the _________________ from an amino acid.

5-11: Decarboxylase Test

A

Carboxyl group

110
Q

Enzymes called deaminases catalyze the removal of the ________________ from an amino acid.

5-11: Decarboxylase Test

111
Q

What is meant by the substrate of an enzyme?

5-11: Decarboxylase Test

A

The substrate of an enzyme is amino acid that has its variable group removed during the reaction resulting in the production of an amine or carboxyl group being released

112
Q

Decarboxylases catalyze reactions that produce ________________ (pH) products.

5-11: Decarboxylase Test

113
Q

The pH indicator bromocresol purple is ______________ at pH 6.8 and above and ______________ below pH 5.2.

5-11: Decarboxylase Test

A

Purple
Yellow

114
Q

Why is mineral oil added to the decarboxylase broth tubes before incubation?

5-11: Decarboxylase Test

A

To seal off from oxygen and encourage fermentation

115
Q

Why does the decarboxylase-positive tube have to turn yellow before it can turn purple?

5-11: Decarboxylase Test

A
  • Accumulation of acidic end products will occur first
  • Then low pH organisms will react to acidic enviornment and either produce decarboxylases or not resulting in positive (purple) or negative (yellow) results
116
Q

Describe a positive result in this experiment and describe a negative result. What kind of pH is present in a + result and is present in a - result?

5-11: Decarboxylase Test

A

Positive: Purple, Alkaline, decarboxylases resulted in an increase in pH
Negative: Yellow, Acidic, lack of decarboxylases meant pH remained negative

117
Q

Why is glucose included in the decarboxylase broth?* (Hint: answer is not “as a food source”)

5-11: Decarboxylase Test

A
  • Low glucose concentration promotes fermentation lowering pH and causing proteases to activate (decarboxylase)
118
Q

This decarboxylation test cannot be performed successfully on organisms that cannot ferment glucose. Explain why this is the case.*

5-11: Decarboxylase Test

A
  • If the organism cannot lower the pH through fermentation the proteases will not be activated and no color change will occur
119
Q

Cadaverine and putrescine are examples of compounds known as __________________.

5-11: Decarboxylase Test

120
Q

The names, cadaverine and putrescine, suggest that the compounds might be found in _______________, which they are, as a result of the decarboxylation of ________________ (what molecule) by bacteria.

5-11: Decarboxylase Test

A
  • Rotting Flesh
  • Lysine -> Cadaverine
  • Ornothine -> Putrescine
121
Q

What is phenylalanine (what type of monomer is it)?

5-12: Phenylalanine Deaminase

A
  • Amino Acid substrate
122
Q

Phenylalanine is part of an ingredient found in the popular artificial sweeteners known as _________________ or ___________________.

5-12: Phenylalanine Deaminase

A

Aspartane
Nutrasweet

123
Q

The enzyme that some bacteria can produce that removes an amino group from phenylalanine is called ___________________________________.

5-12: Phenylalanine Deaminase

A

Phenylalanine Deaminase

124
Q

The reagent that is added to bacterial growth on phenylalanine deaminase agar in order to determine if the enzyme has been produced, is __________________________.

5-12: Phenylalanine Deaminase

A

Ferric Chloride

125
Q

Describe a positive result in the experiment and a negative result.

5-12: Phenylalanine Deaminase

A
  • Green indicates positive as Phenolpyruvate is present
  • Yellow indicates negative as no phenolpyruvate is present
  • Note this is not a pH test as the phenyl pyruvate and ammonium cancel eachother’s pH
126
Q

Phenylalanine is broken down to _____________________ and _____________________.

5-12: Phenylalanine Deaminase

A

Ammonium
Phenylpyruvic Acid

127
Q

Why can’t you use a pH indicator to detect phenylalanine deamination?

5-12: Phenylalanine Deaminase

A

Because the Phenylpyruvic acid (acid) and Ammonium (base) cancel eachother’s pH

128
Q

A green color results in this experiment if _______________________________ reacts with the reagent ________________________.*

5-12: Phenylalanine Deaminase

A

Phenylpyruvic Acid
Ferric Chloride

129
Q

Like all amino acids, phenylalanine is a source of the two elements ______________ and _____________ for the bacteria.

5-12: Phenylalanine Deaminase

A

Nitrogen
Carbon

130
Q

Write the chemical formula for an amine group (amino group).

5-12: Phenylalanine Deaminase

131
Q

What is the difference between the deamination and the decarboxylation of an amino acid (Hint: think of oxygen requirements)?

5-12: Phenylalanine Deaminase

A

Decarboxylation occurs in anaerobic conditions
Deamination occurs in aerobic conditions

132
Q

Urea can be hydrolyzed to carbon dioxide and ___________________ by bacteria that produce the enzyme ______________________________.

5-18: Urease Test

A

Ammonium
Urease

133
Q

Urinary tract pathogens from the genus ______________________ can be distinguished from other enteric bacteria by their rapid urease activity.

5-18: Urease Test

134
Q

What other disease is caused by this organism? (Proteus)

5-18: Urease Test

A

Proteus mirabilis and Proteus vulgaris cause diaper rash
H. Pylori causes stomach ulcers

135
Q

Urease-positive organisms break down urea to _______________, which ________________ the pH of the medium and changes the color of the pH indicator phenol red from _______________ to _______________.

5-18: Urease Test

A

Ammonium
Increases
Orange
Pink

136
Q

Describe a positive urease test and a negative test (include the relative pH).

5-18: Urease Test

A

Pink: Urease was produced leading to release of ammonium as urea broke down (+), pH >8.4
Orange: (-) pH <8.4
Yellow: Organism was unable to produce urease and did not break down urea (-), pH <8.4

137
Q

In order for a urease-positive organism to turn the urea broth pink, the organism must ___________________ the pH of the medium enough to overcome the effects of the __________________, potassium phosphate.

5-18: Urease Test

A

Increase
Buffer

138
Q

The urease test is an important part of the identification of the organism _________________________ , that causes peptic ulcers.

5-18: Urease Test

139
Q

How does this organism (H. pylori) survive stomach acid?

5-18: Urease Test

A

Rapid Urease production leads to ammonium production which can neutralize the stomach acid

140
Q

A rapid urease test is performed on the ______________________ of patients with peptic ulcer symptoms.

5-18: Urease Test

141
Q

Why is it important that urease broth contain a buffer that can resist alkalinization of the medium from peptone metabolism?*

5-18: Urease Test

A

To avoid false positives from reversion of peptone metabolism

142
Q

Ammonia is used by many bacteria as a source of the element ______________________.

5-18: Urease Test

143
Q

Bacteria use nitrogen, in order to make what two classes of macromolecules?*

5-18: Urease Test

A

Nucleic acids and Polypeptides

144
Q

TSI is a differential medium that is useful in differentiating _______________________________________.

2-7: Triple Sugar Iron (TSI) agar

A

Fermentation

145
Q

Triple Sugar Iron agar contains 3 sugars in its recipe. The monosaccharide ___________ is supplied in a low percentage (0.1%) while the disaccharide sugars ____________ and ____________ are provided in a higher percentage (1%).

2-7: Triple Sugar Iron (TSI) agar

A

Glucose
Lactose
Sucrose

146
Q

The results of TSI tubes are recorded according to the pH of the __________ over the pH of the ________(e.g., K/A). The pH indicator in TSI is __________________ which is the color ______________ when it’s acidic and ______________ when it’s alkaline.

2-7: Triple Sugar Iron (TSI) agar

A

Slant
Butt
Phenol Red
Yellow
Red

147
Q

Fermenters only capable of fermenting _______________ will run out of sugar within the first few hours during incubation. Explain what these organisms do next and what the results look like in both the slant and butt.

2-7: Triple Sugar Iron (TSI) agar

A

Glucose
- These organisms will then swap to peptone digestion which will turn the slant red while the anaerobic butt remains yellow

148
Q

Fermenters capable of fermenting one or both disaccharides (sucrose and/or lactose) will never run out of sugar during the required 24-hour incubation. Explain what TSI slant looks like in this case.

2-7: Triple Sugar Iron (TSI) agar

A
  • Because Fermentation will occur throughout the entire 24H period the slant and butt will both be yellow
149
Q

Besides sugar, other substrates found in TSI agar include _____________ and ________________________. These substrates are a source of reducible ____________, and digestion of these substrates leads to the formation of H2S.

2-7: Triple Sugar Iron (TSI) agar

A

Cysteine
Sodium Thiosulfate
Sulfur

150
Q

H2S is a colorless gas that smells like rotten eggs. It is visually detected when a black color (precipitate) forms by reacting with the iron-containing ingredient ___________________________________.

2-7: Triple Sugar Iron (TSI) agar

A

Ferrous ammonium sulfate

151
Q

H2S can be made by certain species of sulfur-reducing bacteria. The formation of H2S requires a reducing environment where excess hydrogen is present, and the pH is _____________.

2-7: Triple Sugar Iron (TSI) agar

152
Q

Explain why a K/K [H2S +] result is not possible.

2-7: Triple Sugar Iron (TSI) agar

A

Because in order for the agar to be H2S+ it would require an acidic environment which is not possible if the tube is K/K

153
Q

Enzymes that are released into the bacterial environment and work there are known as ______________________________.

2-7: Triple Sugar Iron (TSI) agar

A

Exoenzymes

154
Q

Bacteria that produce protein-digesting (proteolytic) enzymes break down large ___________________ to smaller _____________________ or individual ____________________.

5-16: Casease Test

A

Proteins
Peptides
Amino Acids

155
Q

Why do bacteria have to break down proteins in their environment (outside the cell) in order to use the proteins as a food source?

5-16: Casease Test

A

Because it is too big to be taken across the membrane and therefore needs to be broken down.

156
Q

Why is milk opaque?* Why does the milk agar turn clear when an organism produces the casease enzyme?*

5-16: Casease Test

A

Casein is the protein that gives milk its white color but when it is broken down it becomes clear and less opaque

Top line is Casease +

157
Q

What is a polypeptide? What is the difference between a polypeptide and a peptide? Between a polypeptide and a protein? Between a peptide and an amino acid?

5-16: Casease Test

A
  • A polypeptide is a long chain of amino acids
  • A peptide is a shorter chain of amino acids
  • A protein can be made of multiple polypeptides
  • An amino acid is a monomer.
158
Q

What is the difference between casein and pancreatic digest of casein?

5-16: Casease Test

A
  • Casein is large bulky protein
  • Pancreatic digest of casein has been broken down and is an accessible food source
159
Q

Why are whole, undigested proteins (as opposed to peptones), not usually included in the microbiological growth media?*

5-16: Casease Test

A

Because they’re often too large or hard to break down if the bacteria dont have the right exoenzymes. Food sources must be able to be digested and taken into the cell

160
Q

Most nitrate-reducing bacteria contain the enzyme ________________________ and perform a single-step reduction of nitrate.

5-8 Nitrate Reduction

A

Nitrate Reductase

161
Q

What is denitrification?

5-8 Nitrate Reduction

A

The loss of biologically available nitrogen; occurs by release of nitrogenous gas

162
Q

Write the chemical formulas for nitrate, nitrite, potassium nitrate, ammonia, ammonium ion, molecular nitrogen, and nitric oxide.

5-8 Nitrate Reduction

A

Nitrate: NO3
Nitrite: NO2
Potassium Nitrate: KNO3
Ammonia: NH3
Ammonium ion: NH4+
Nitrogen: N2
Nitric Oxide: NO

163
Q

Why is the conversion of nitrate to nitrite an example of reduction?

5-8 Nitrate Reduction

A

Because you go from NO3 to NO2 which is a gain in electrons for nitrogen as nitrogen’s oxidation state goes from +5 to +3 (oxidation state going down is reduction)

164
Q

Why is the conversion of nitrite to the ammonium ion an example of reduction?

5-8 Nitrate Reduction

A

Because you go from NO2 to NH4+ and the oxidation state goes from +2 to -3 which is reduction

165
Q

Why do bacteria need nitrogen (which 2 macromolecules require nitrogen)? (2 answers)*

5-8 Nitrate Reduction

A

Nucleic acids and Peptides

166
Q

Why are there Durham tubes in the nitrate broth?

5-8 Nitrate Reduction

A

Because if denitrification occurs, N2 gas will be produced

167
Q

Explain thoroughly each of the following results in this experiment.
a. After incubation, turbid nitrate broth turns red when test reagents A and B are added.
b. After incubation, turbid nitrate broth does not turn red when reagents A and B are added.
c. A turbid tube that did not turn red when reagents A and B were added turns red when zinc is added.
d. A turbid tube that did not turn red when reagents A and B were added does not turn red when zinc is added.
e. There is gas in the Durham tube after incubation with a non-fermenting organism.

5-8 Nitrate Reduction

A

A. (+) Nitrate is present and bacteria did reduce nitrate
B. Zinc must be added to determine if zinc can reduce nitrate (-) or if no nitrite is present (+). Either no reduction or full reduction occured
C. (-) Red indicates bacteria did not reduce, the zinc did
D. (+) Bacteria were able to reduce nitrate broth and either underwent ammonification or underwent full nitrification
E. (+) Full denitrification occured resulting in gas in durham tube

168
Q

Describe the results that would occur in an uninoculated control tube when reagents A and B are added to it?* Why?*

5-8 Nitrate Reduction

A

The tube would stay transparent as there is no nitrite to react with

169
Q

What results should an uninoculated control tube give when ONLY zinc is added to it?* Why?*

5-8 Nitrate Reduction

A

The broth should remain transparent as the zinc is for reducing the broth but is not an indicator

170
Q

What results should an uninoculated control tube give when reagents A, B, and zinc are added to it?* Why?*

5-8 Nitrate Reduction

A

The Tube will turn red because the zinc will reduce nitrate and the indicator will show the presence of nitrite

171
Q

What do reagents A and B test for? What does the zinc test for?

5-8 Nitrate Reduction

A
  • Reagents A and B test for nitrite presence
  • Zinc test if reduction occured and will turn nitrate into nitrite if it has not already occured
172
Q

Why is the conversion of nitric oxide to molecular nitrogen an example of reduction?

5-8 Nitrate Reduction

A

Because the electronegativity went from +2 to 0

173
Q

After incubation, gas in the Durham tube in a tube inoculated with a non-fermenting organism means that _____________________ has occurred. Is this a + or a – test for nitrate reduction? What type of gas has been produced?

5-8 Nitrate Reduction

A

Denitrification
Positive
Nitrogen

174
Q

What is the original source of nitrate in the nitrate broth? Write the formula for this compound.

5-8 Nitrate Reduction

175
Q

What are aerobic bacteria? (see textbook)

5-6 Catalase Test

A

Bacteria that require an oxygenated environment because they use oxygen as their final electron acceptor

176
Q

What are aerotolerant bacteria? (see textbook)

5-6 Catalase Test

A

Bacteria that can grow in an oxygenated environment but prefer an anaerobic environment

177
Q

What are facultative bacteria? (facultative anaerobes) (see textbook)

5-6 Catalase Test

A

Facultative anaerobes can grow better with oxygen but still grow at a slower rate in an anaerobic environment

178
Q

Aerobic and aerotolerant bacteria produce enzymes capable of detoxifying 2 very potent cellular toxins, ___________________________ and __________________________.

5-6 Catalase Test

A

Superoxide dismutase
Catalase

179
Q

The enzyme _____________________ catalyzes conversion of ______________________, the more lethal of the 2 compounds, to _________________________.

5-6 Catalase Test

A

Superoxide dismutase
Superoxide radicals
Hydrogen peroxide

180
Q

The enzyme __________________ converts hydrogen peroxide to _____________ and _________________.

5-6 Catalase Test

A

Catalase
Water
Oxygen

181
Q

What happens when hydrogen peroxide is added to a catalase-positive culture? What is the appearance of the catalase-negative culture when hydrogen peroxide is added?

5-6 Catalase Test

A
  • The culture will fizz and bubble
  • A catalase negative culture will not bubble when it encounters hydrogen peroxide
182
Q

This test is most commonly used to differentiate members of the catalase-positive __________________________ from the catalase-negative __________________________.

5-6 Catalase Test

A

Micrococcaceae
Streptococcaceae

183
Q

If an organism is catalase-negative, what does that tell you about its oxygen requirements?* Why?*

5-6 Catalase Test

A

It is anaerobic because it does not have a way to get rid of superoxide radicals

184
Q

If an organism is catalase-positive, what does that tell you about its oxygen requirements?* Why?*

5-6 Catalase Test

A

It is aerobic as it has a way to get rid of harmful superoxide radicals

185
Q

What is the morphology and gram reaction of the organism used in this experiment? Even before you performed the catalase test on this organism, you should suspect that it is catalase positive because:

5-6 Catalase Test

A

S. Epidermidis (Gram positive coccus) which lives on the skin and is an aerobe.

E. Faecalis (Gram positive coccus) which is found in the colon where there isn’t much oxygen.

186
Q

Would you expect bacteria that are facultative anaerobes to be catalase-positive or catalase-negative?* Why?*

5-6 Catalase Test

A

They should be catalase positive because using oxygen as an electron acceptor will lead to superoxide radical productions. This means bacteria must have a way to deal with them

187
Q

The oxidase test that is useful in the presumptive identification of the gram-negative coccus _________________________. Name 2 diseases caused by species of this genus.

5-7 Oxidase Test

A

Neisseria
N. meningitidis causes meningitis
N. gonorrhoeae causes gonorrhea

188
Q

Bacteria that belong to the family Pseudomonadaceae are oxidase __________________; bacteria that belong to the family Enterobacteriaceae are oxidase ________________________.

5-7 Oxidase Test

A

Positive
Negative

189
Q

What does a positive oxidase test look like? Include the time frame.

5-7 Oxidase Test

A

A positive test will result in a red smear after 20 seconds

190
Q

What does a negative oxidase test look like? (2 answers)*

5-7 Oxidase Test

A

No color change within 20sec
Color change (pink)after a minute due to reagent oxidation

191
Q

If your bacteria turn pink after more than one minute has elapsed since the oxidase reagent was added, why is this happening?

5-7 Oxidase Test

A

The reagents are unstable and were oxidized independently given enough time

192
Q

The quantitative sensitivity disk method of antibiotic sensitivity testing is called the _________________
method.

7-3 Antibiotic Sensitivity Testing (Kirby-Bauer Method)

A

Kirby-Bauer

193
Q

In this method, the diameter of the ___________________________ around the disk is measured to the
nearest _____________________ (unit).

7-3 Antibiotic Sensitivity Testing (Kirby-Bauer Method)

A

Zone of inhibition
Milimeters

194
Q

The inhibition zone diameter that is produced in this experiment will indicate the ___________________ of a bacterium to each antibiotic.

7-3 Antibiotic Sensitivity Testing (Kirby-Bauer Method)

A

Sensitivity

195
Q

How do you determine if an organism is susceptible to a particular antibiotic in this test? How do you determine if the organism is resistant?

7-3 Antibiotic Sensitivity Testing (Kirby-Bauer Method)

A

The susceptibility zone is measured and compared to a document that states according to what antibiotic is being used a diameter under X is resistant and over X is susceptible

196
Q

What will a negative test for antibiotic sensitivity look like? Describe such a result and explain what it means.

7-3 Antibiotic Sensitivity Testing (Kirby-Bauer Method)

A

A negative would be shown as no ZOI or a small ZOI. This means that the antibiotic was unable to stop or kill the bacteria

197
Q

What will a positive test for antibiotic sensitivity look like? Describe such a result and explain what it means.

7-3 Antibiotic Sensitivity Testing (Kirby-Bauer Method)

A

A large ZOI that is greater than the susceptibility break point
ex. Breakpoint <10mm, actual D = 18mm

198
Q

Why is it important that the sensitivity disks used in this experiment contain a specific, standardized amount of antibiotic?* How is this test standardized (i.e. what is standardized in this test)?(4)

7-3 Antibiotic Sensitivity Testing (Kirby-Bauer Method)

A
  • To ensure all results are reliable and safe for humans
  • The agar is 4mm (thinner than usual) to promote lateral diffusion
  • pH is kept to 7.2-7.4 to mimic blood pH
  • Agar is incubated at 37 degrees C to mimic human body temp
  • Agar is incubated for 18 hours to select for younger bacteria
199
Q

Why is it important that the results of antibiotic sensitivity test guide treatment of bacterial infections?

7-3 Antibiotic Sensitivity Testing (Kirby-Bauer Method)

A

It allows us to understand how different drugs interact with bacteria and what is an effective medication that could be used to adress a bacterial infection appropriately

200
Q

Penicillin is an antibiotic produced by the mold ________________________ (genus), which is also responsible
for __________________________ (type of food).

7-3 Antibiotic Sensitivity Testing (Kirby-Bauer Method)

A

Penicillium
Cheese

201
Q

What is “intermediate susceptibility” to an antibiotic?* Under what conditions would it be appropriate to use an antibiotic to which a particular organism was intermediately susceptible?*

7-3 Antibiotic Sensitivity Testing (Kirby-Bauer Method)

A
  • When the ZOI is above resistant but below susceptibility
    ex. R<5, I=6-8, S>9
  • Appropriate if the person is allergic to other antibiotics or can be used with another drug for enhanced effects
202
Q

True or false: All bacteria found within the clear area around the disks are dead. Why did you answer true or false?

7-3 Antibiotic Sensitivity Testing (Kirby-Bauer Method)

A

False. There are bacteriostatic antibiotics which are not bacteriocidal. Often times with gram negative infections when they are lysed they release lipid-A which is toxic and can cause shock and even death. Therefore a bacteriostatic drug is better to pause development and allow the patient’s body to fight the infection in a safer way

203
Q

What is a broad-spectrum antibiotic? When would it be better to use a broad-spectrum antibiotic? Why?

7-3 Antibiotic Sensitivity Testing (Kirby-Bauer Method)

A

A broad-spectrum antibiotic is an antibiotic that many bacteria are susceptible both gram positive and gram negative. Broad spectrum antibiotics are generally reserved as a last ditch effort as it also kills natural flora. Otherwise targetted antibiotics are usually more appropriate

204
Q

What is a narrow spectrum antibiotic? When would it be better to use a narrow spectrum antibiotic? Why?

7-3 Antibiotic Sensitivity Testing (Kirby-Bauer Method)

A

Narrow spectrum antibiotics are antibiotics that are effective at targetting a specific bacteria. This is effective when you know exactly what the bacterial infection is and will not affect your natural flora

205
Q

What is drug synergy? What does plate look like if two drugs are synergistic?

7-3 Antibiotic Sensitivity Testing (Kirby-Bauer Method)

A
  • Drug synergy is when antibiotics combine and have a greater effect together than their individual effects summed
  • If the drugs synergize the ‘halo’ between the two drugs will be bigger than the halo around the drug itself
206
Q

Name the causative agent (organism) of amoebic dysentery (amebiasis).

12-3: Examination of Common Protozoans of Clinical Importance

A

Entamoeba histolytica

207
Q

Amoebic dysentery is most common in areas with __________________________.

12-3: Examination of Common Protozoans of Clinical Importance

A

Poor water sanitation

208
Q

Identification of the amoebic dysentery organism may be made by finding either trophozoites or _________________ in a _________________________.

12-3: Examination of Common Protozoans of Clinical Importance

A

Cysts
Stool Sample

209
Q

Infection with the amoebic dysentery organism occurs when a human host ingests _______________ (stage) in _____________________________________.

12-3: Examination of Common Protozoans of Clinical Importance

A
  • Cyst
  • Contaminated Food or Water
210
Q

__________________ of the amoebic dysentery organism are shed in the feces and may _______________________.

12-3: Examination of Common Protozoans of Clinical Importance

A
  • Cyst
  • Be infective
211
Q

Cysts of the amoebic dysentery protozoan may persist in the original host, resulting in ____________________________________.

12-3: Examination of Common Protozoans of Clinical Importance

A

Asymptomatic Carriers

212
Q

How does the organism that causes amoebic dysentery move through its environment?

12-3: Examination of Common Protozoans of Clinical Importance

A

Psuedopods

213
Q

The only ciliated human pathogen is (genus and species) __________________________________ and it causes the disease __________________________________.

12-3: Examination of Common Protozoans of Clinical Importance

A
  • Balantidium coli
  • Balantidiasis
214
Q

____________________ in sewage-contaminated water are the infective form of Balantidium coli.

12-3: Examination of Common Protozoans of Clinical Importance

215
Q

Most infections of Balantidium coli are asymptomatic, however it can cause the following symptoms in some patients:

12-3: Examination of Common Protozoans of Clinical Importance

A

Diarrhea (bloody,invasive)

216
Q

Name the organism that causes giardiasis.

12-3: Examination of Common Protozoans of Clinical Importance

A

Giardia lamblia (intestinalis)

217
Q

Transmission of giardiasis typically involves:

12-3: Examination of Common Protozoans of Clinical Importance

A

Fecal or oral transmission due to drinking river/lake water

218
Q

What is the infective stage of the organism that causes giardiasis?

12-3: Examination of Common Protozoans of Clinical Importance

219
Q

Diagnosis of giardiasis is made by identification of the protozoan trophozoites or ______________ in ____________________.

12-3: Examination of Common Protozoans of Clinical Importance

A

Cysts
Stool Sample

220
Q

The STI/STD known as trichomoniasis is more common in ________________ than _________________.

12-3: Examination of Common Protozoans of Clinical Importance

221
Q

Protozoal vulvovaginitis in humans is caused by (genus and species) ______________________________.

12-3: Examination of Common Protozoans of Clinical Importance

A

Trichomonas Vaginalis

222
Q

Trichomonas is typically transmitted by _______________.

12-3: Examination of Common Protozoans of Clinical Importance

A

Sexual transmission

223
Q

The ____________________ is the only stage of the Trichomonas life cycle and moves by _______________________.

12-3: Examination of Common Protozoans of Clinical Importance

A

Trophozoite
Swimming

224
Q

What drug is used to cure (treat) Trichomonas, Entamoeba, Balantidium, and Giardia?

12-3: Examination of Common Protozoans of Clinical Importance

A

Flagyl
(metronidazole)

225
Q

The genus of the organism that causes African trypanosomiasis, a disease commonly known as ______________________________, is __________________________________.

12-3: Examination of Common Protozoans of Clinical Importance

A

African Sleeping Sickness
Trypanosoma

226
Q

The intermediate host (vector) for African trypanosomiasis is the _______________________.

12-3: Examination of Common Protozoans of Clinical Importance

A

Tsetse fly

227
Q

The stage of Trypanosomes that multiplies in the intermediate host is the ___________________, while the ___________________ stage multiplies in the human host.

12-3: Examination of Common Protozoans of Clinical Importance

A

Epimastigote
Trypomastigote

228
Q

Which area or part of the body is affected during the advanced stages of infection that lead to coma and death in African Sleeping Sickness?

12-3: Examination of Common Protozoans of Clinical Importance

229
Q

Name a disease caused by Trypanosoma other than African sleeping sickness.* How is this disease transmitted?* In what part of the world is the disease most common?*

12-3: Examination of Common Protozoans of Clinical Importance

A

South America Sleeping Sickness
Transmitted by bug bite
South America

230
Q

Name the genus and species of the 4 protozoans that cause malaria.

12-3: Examination of Common Protozoans of Clinical Importance

A

Plasmodium falciparum
Plasmodium malariae
Plasmodium ovale
Plasmodium vivax

231
Q

The most severe form of malaria involves which organism? (genus and species)

12-3: Examination of Common Protozoans of Clinical Importance

A

Plasmodium falciparum

232
Q

The arthropod vector for malaria is the _____________________________________.

12-3: Examination of Common Protozoans of Clinical Importance

A

Anopheles Mosquito

233
Q

For malaria, the ________________________ stage of the pathogen is introduced into a human by a bite by the arthropod vector.

12-3: Examination of Common Protozoans of Clinical Importance

A

Sporozoite

234
Q

For malaria, the _____________________ stage are released from the ruptured liver cells in humans, enter the blood, and infect erythrocytes.

12-3: Examination of Common Protozoans of Clinical Importance

A

Merozoites

235
Q

Name two drugs in current use in the treatment of malaria.*

12-3: Examination of Common Protozoans of Clinical Importance

A

Quinine, chloroquine

236
Q

The sexual phase of the life cycle of Toxoplasma occurs in the _______________ where ____________________ are produced and shed in the ____________________.

12-3: Examination of Common Protozoans of Clinical Importance

A

Cat intestines
Oocysts
Feces

237
Q

The more serious form of toxoplasmosis involves:

12-3: Examination of Common Protozoans of Clinical Importance

A

An infection of a fetus across the placenta that can result in still birth

238
Q

Fetal infection with toxoplasmosis may result in:

12-3: Examination of Common Protozoans of Clinical Importance

A

Still birth

239
Q

______________________ patients may suffer fatal complications from toxoplasmosis.

12-3: Examination of Common Protozoans of Clinical Importance

A

Immunocompromised or pregnant

240
Q

Give 2 reasons why sodium sulfite and basic fuchsin are included in Endo agar.

8-12 Endo Agar

A
  • They inhibit the growth of gram-positive bacteria
  • They both test for lactose fermentation
241
Q

Endo agar inhibits the growth of _______________________ bacteria.

8-12 Endo Agar

A

Gram positive

242
Q

The fermentable carbohydrate in Endo agar is _____________________.

8-12 Endo Agar

243
Q

What will colonies of lactose fermenters look like growing on Endo agar?

8-12 Endo Agar

A

Lactose fermentation -> Acids -> If rapid fermenter green metalic sheen, If slow fermenter dark pink/red

244
Q

What appearance is created in this agar by lactose-fermenting organisms that produce large amounts of acid?

8-12 Endo Agar

A
  • A metalic sheen green on the top of the agar and black appearance from the back of the plate
245
Q

Name one coliforms (genus and species) that produce large amounts of acids when grown on Endo agar.

8-12 Endo Agar

246
Q

What appearance will be created by lactose non-fermenters growing on Endo agar?

8-12 Endo Agar

A

Light pink/colorless growth

247
Q

Bacteria that will grow on Endo agar are Gram _______.

8-12 Endo Agar

248
Q

Endo agar is used to detect fecal contamination in water and dairy products. What is the connection between fecal contamination and the bacteria that produce a metallic sheen on this agar?*

8-12 Endo Agar

A

Water and milk are tested for because E.coli lives in the intestines and their presence signals fecal contamination. The metalic sheen growth on endo agar is a visual indicator that the bacteria was a lactose fermenter and is presumptively a fecal coliform

249
Q

If an organism that cannot ferment lactose grows on Endo agar, what is it using as a food source?* What kind of pH would this organism be expected to produce in the medium?* What will the colonies of this kind of organism look like?*

8-12 Endo Agar

A

Peptones, this organism would have a alkaline pH from peptone digestion. This organism would grow in light pink or colorless colonies

  • It is important to note, nonfermenter could still be possible salmonella or shigella
250
Q

What will be the appearance of a presumptive coliform organism growing on this agar? Why?

8-12 Endo Agar

A
  • Metalic sheen green if the organims is a rapid fermenter of lactose
  • Dark pink/red if the organism is a slow fermenter of lactorse
251
Q

If an organism will not grow on Endo agar, that is a presumptive indication that the organism is __________________.

8-12 Endo Agar

A

Gram Positive

252
Q

Helicobacter pylori

A

Urease positive - causes ulcers

253
Q

Pseudomonas aeruginosa

A

Gelatinase positive and strict (obligate) aerobe - causes skin infections in burn patients, lung infections in cystic fibrosis patients

254
Q

Staphylococcus aureus

A

G+, Osmotolerant and facultative anaerobe - causes pimples, boils, etc.

255
Q

Clostridium perfringens

A

Strict (obligate) anaerobe - causes gangrene

256
Q

Neisseria gonorrhoeae

A

G-, Microaerophile - causes gonorrhea

257
Q

Listeria monocytogenes

A

Psychrotroph - causes listeriosis

258
Q

Bacillus stearothermophilus

A

G+, Thermophile and used in Kilit ampule

259
Q

Bacillus

A

Most UV resistant when endospores are formed

260
Q

Escherichia coli

A

G-, Mesophile and metallic green on Endo agar - causes UTI and sepsis

261
Q

Klebsiella pneumoniae

A

Red on Endo agar

262
Q

Salmonella typhi

A

Clear/light pink on Endo agar - causes typhoid fever

Microbiology 210 (12 decks)

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