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Pathogenic Microbiology > Lab Test 1 > Flashcards

Lab Test 1 Flashcards

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1
Q

What is the Kinyoun Stain used for

A

Acid-fast species of the bacterial genus Mycobacterium. It is a variation of a method developed by Robert Koch in 1882

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2
Q

Which was the Mordant used in the Acid-fast stain

A

DMSO

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3
Q

In our lab what experiment did we use: M. Smegmatis & S. epidermidis

What were the results

Describe appearance of each

A

Acid-fast stain

Staph epidermis = Not acid fast (Cocci Blue)

Mycobacterium smegmatis = Acid fast. (Pink Bacillus)

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4
Q

Why is DMSO used in fast-acid staining?

Which stain does it allow to penetrante & attach to the Waxy Cell Wall?

A

It is a Surfacent and allows Carbol Fuchsin to penetrante the Waxy cell wall

Carbol Fuchsin

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5
Q

Which type of bacteria are associated with Acid-fast cells

Why?

A

Mycobacteria

They have a Waxy Cell Wall

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6
Q

A Positive indication for Acid-Fast bacteria is….

A negative is

A

Positive = Red / Pink

Negative = Blue

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7
Q

Name the alcohol used in Acid Fast staining

Name the alcohol used in gram staining

A

Acid fast = Acid Alcohol

Gram staining = Aceton Alcohol

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8
Q

2 stains used in Acid Fast in Order

Are they basic or acidic

Positive or Negative charged

Reagent

Decolorizer

A

Carbol Fuchsin

Methylene Blue

Basic

DSMO

Acid Alcohol

Positive charged

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9
Q

How long does the DMSO-carbol fuchsin stay on the slide

A

4 minutes

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10
Q

How many passes with the Acid Alcohol in the Acid-Fast decolorizing technique

How many passes with the Aceton Alcohol in the Gram-staining Tech

A

1 pass

2 passes

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11
Q

The fast Acid cells are described as “pleomorphic”

True or False

A

True

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12
Q

When do you blot slides?

A

At the very end

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13
Q

Name the reagents used for gram-staining in order.

How long do you leave each on for?

A
  1. Crystal Violet 1 min (Primary Stain)
  2. Grams (Lugols) Iodine 1 minute (Mordant)
  3. Acetone alcohol (Decolorizer) Remove instantly
  4. Safranin 1 minute (Counter Stain)
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14
Q

Decolorizer, decoloizes….

A

Gram-negative & non acid fast

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15
Q

In the Gram Staining experiment which 2 bacteria were looked at?

What were the results?

A

Staphylococcus epidermidis = Gram Positive (Purple Grapes)

Escherichia coli = Gram Negative (Red Rods)

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16
Q

Name of paper for drying slide

A

Bibulous paper

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17
Q

Wet mount 4 steps

A
  1. Loopful of culture
  2. 1 drop of stain
  3. Cover slip
  4. View with 4, 10, 40x
    NO Oil Immersion
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18
Q

Difference between Iris Diaphragm & Condensor lens

A

Iris Allows more light

Condenser Focuses it

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19
Q

What does parfocal mean

A

Ability to switch magnification without readjusting

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20
Q

2 physical characteristics of Corynebacterium

A

Rod shapped bacillus

Palisades arrangements

Gram-positive

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21
Q

Staphylococcus is gram-negative or gram-positive?

Describe shape and arrangement/ color after gram stain

A

Gram-positive

Round clusters like grapes

Purple

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22
Q

What are chromophores

A

Color part of stain

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23
Q

Difference between basic and acidic dyes

A

Basic dyes have a positive charge and adhere to the cell (due to the cell having a negative charge. )

Acidic dyes have a negative charge and stain the background (allowing the cell to be seen)

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24
Q

Crystal Violet is an example of what kind of stain?

A

Simple stain

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25
Q

Where are fusiform bacteria found in humans

A

Mouth

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26
Q

Can you ID bacterial species using a Simple Stained slide?

A

No, simple stain only IDs the morphology

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27
Q

How do we store and label petri dishes?

Why?

A

Label and store “upside-down”

To prevent condensation

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28
Q

Disinfectants are effective against which type of pathogens?

Which are they not effective against?

A

Vegetative cells & viruses

Not effective against ENDOSPORES

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29
Q

Which is our disinfectant in the laboratory?

A

Vesphene

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30
Q

This type of medium allows for growing of isolated colonies and are easy to observe

Disadvantage: they can dry out easily

A

Agar plates

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31
Q

These allow for growing large number of bacteria and at varying oxygen levels

Disadvantage: you cannot isolate them

A

Broths

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32
Q

These dry out less easily than agar palates and take up less space.

But have less visibility than agar plates

A

Slants

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33
Q

Retrieving a culture from a tube with a loop, the cap of the tube is removed and held with the fingers from which hand?

A

The one with the loop

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34
Q

Kinyoun Method is for what

A

Acid fast / Mycobacterium

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35
Q

The Schaeffer-Fulton method is also called…

A

(Endo)Spore Stain

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36
Q

How long do we let the Methylene Blue stay on the Kinyoun Method?

A

30 seconds

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37
Q

The endospore species is…

Where is it found? Aerobic ?

Gram-negative or gram-positive

Morphology and arrangements

A

Bacillus Cereus

Soil/ Yes

Gram-positive

Streptobacillis

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38
Q

After we air-dry and heat fix the endospore slide we add Malachite Green and then

A

Set on heated hot plate for 5 Minutes

39
Q

How long do we let the malachite green cool after we take it off the 2nd hot plate?

A

3 minutes

40
Q

What is the stain used in “negative stain”

A

Nigrosin

41
Q

With the negative stain what were the 2 species we examined?

What was the alternative choice

A

B. Cereus & S. Epidermidis

Mouth biofilm

42
Q

What steps do we skip in both the negative stain and the capsule stain?

A

Heat fixing

Rinsing with distilled water

43
Q

Which is species is in the purple broth?

Which staining method is it used for?

Gram positive or negative

Morphology and arrangements

A

Enterobacter aerogenes / Klebsiella aerogenes

Capsule Stain

Gram negative

Diplobacillus

44
Q

Which stain do we use Aqueous Crystal Violet for? 1 minute

A

Capsule stain

45
Q

In the capsule stain we use which three reagents and for how long?

A

Nigrosin

Aqueous Crystal Violet 1 minutos

20% Copper Sulfate 3 minutos

46
Q

What do we use to rinse the slide in the capsule stain?

A

20% Copper sulfate

47
Q

Acid fast bacteria are also (gram-negative/ gram-positive)

A

Gram-positive

48
Q

Name the 2 reagents used in endospore staining, in order

When do we add them

Do we rinse, if so with what?

A

after air dry and heat fixing, place on hot plate and add Malachite Green 5 minutes

Remove from hot plate and rinse with distilled water

Safranin Red & Rinse with distilled water

49
Q

Kinyoun method aka

Reagents

Mordant

Bacteria we looked at, results Color

A

Acid-fast

Basic fuchsin

2ndary stain Methylene Blue

DSMO

Mycobacterium smegmatis = Acid fast Red

S. Epidermidis = non acid fast Blue

50
Q

What are the colors and the species from the Schaeffer-Fulton method.

Name of species used

A

Red = vegetative cells

Green = endospores

Bacillus cereus

51
Q

Exosporium does what what is it associated with?

Composition

A

Protective barrier for endo spore.

Proteins

52
Q

What is the Mordant in the endospore staining

A

Heat

53
Q

What is the 2ndary stain in the Schaeffer-Fulton endospore stain?

A

Safranin

54
Q

Which species doesn’t produce endospores

Clostridium, Mycobacterium, Bacillus

A

Mycobacterium

55
Q

Mycobacterium tuberculosis is visible under which kind of staining?

Results from this stain

A

Kinyoun Acid-fast

Pink

56
Q

Advantage of Kinyoun staining over Ziehl-Neelsen?

Which did we use?

A

Kinyoun doesn’t involve heating releasing poison gas

Kinyoun

57
Q

Name acid-fast genuses

A

Mycobacterium & Nocardia

58
Q

What makes Mycobacterium resistant to staining

A

Mycolic acid

59
Q

Describe E coli

Gram status

Morphology and arrangements

A

Negative (Pink)

Bacilli Single or Diplobacillus

60
Q

Staphylococcus epidermidis before primary stain

A

Colorless

61
Q

Pseudomonas fluorescens after primary stain?

A

Purple

62
Q

Pseudomonas fluorescens

Gram status

Morphology and arrangements

A

Negative

Bacillus

Single

63
Q

Bacillus megaterium after the addition of the Mordant

A

Purple

64
Q

Bacillus megaterium

Gram status

Morphology and arrangements

A

Positive

Streptobacillus

65
Q

Staphylococcus epidermidis after Decolorizer

A

Purple

66
Q

Moraxella catarrhalis after Decolorizer

A

Colorless

67
Q

Moraxella catarrhalis

Gram status

Morphology and arrangements

A

Negative

Diplococci

68
Q

Pseudomonas fluorescens after Decolorizer

A

Clear

69
Q

Bacillus megaterium after counter stain

A

Purple

70
Q

Escherichia coli if you forget to add the counter stain

A

Clear

71
Q

Escherichia coli if you forget to apply the Decolorizer

A

Purple

72
Q

Bacillus megaterium if you forget to add Mordant (Grams Iodine)

A

Red

73
Q

On the pipette pump where is 1ml?

A

At the 0 mark

74
Q

0.1 ml is how much on the pipette pump

A

Between any 2 whole number lines

75
Q

Standard plate count

1 ml from each test tube to the next filled with 9 ml

What is the dilution of A B C

A

A = 1: 100

B = 1: 10,000

C= 1: 1,000,000

76
Q

1 ml from test tube B on the plate. What is the total dilution?

A

1:10,000

77
Q

0.1 ml from testtube B on the plate has What total dilution?

A

1:100,000

78
Q

1 ml from testtube C on the plate has what total dilution?

A

1:1,000,000

79
Q

0.1 ml from test tube C has What total dilution?

A

1:10,000,000

80
Q

What is the control in the standard plate count?

A

1 ml E. Coli

81
Q

Simple staining is…

A

Using 1 dye to stain a bacterial smear

82
Q

Type of attraction between cell and stain

A

Electrostatic

83
Q

Describe shape of Fusiform bacteria

Which group of cell shapes do they fall into?

A

Bacillus

But with tapered ends

84
Q

Staphylococcus aureus, Streptococcus lactis, Bacillus megaterium and Corynebacterium xerosis were examined by this staining type

A

Simple

85
Q

Pleomorphism refers to

A

Demonstrating several different shapes

86
Q

Distinct reddish purple granules with in a cell that show up with Methylene Blue

A

Metachromatic granules

87
Q

3 things you can discern with a good smear

A
  1. Morphology (Rods, Cocci, Shapes)
  2. Arrangement ( Single, Stapho, Strepto)
  3. Internal structures ( endospores & cell inclusions)
88
Q

Goals of a good smear

A
  1. Adhere cells so they are not washed off during the staining and rinsing
  2. Ensure shrinkage doesn’t happen to cells. ( causing Distortion & Artifacts)
  3. Prepare thin smears
89
Q

For solid media when creating a smear which do you add first the smear or the water?

A

Water

90
Q

Where do we write the name of the organism on a slide

A

Left-hand side

91
Q

A hot loop will cause what to happen to the bacteria

A

They will go into the air

92
Q

When do we flame the tube?

A

After opening

After putting the inoculation loop in

93
Q

Protoslo does…

A

Slows movement of bacteria

94
Q

Gram staining is which kind

A

Differential

Pathogenic Microbiology (31 decks)

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