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[3Y1S] MT 6320 > U2.1.1 L : Experiment 6-7 > Flashcards

U2.1.1 L : Experiment 6-7 Flashcards

1
Q

Differential Staining

Principle : used to distinguish if the isolate is Gr (+) or Gr (-)

A

Gram Staining

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2
Q

Gram Staining

Gram (+) bacteria : color

A

stained purple

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3
Q

Gram Staining

Gram (-) bacteria : color

A

stained red

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4
Q

T/F Gram Staining is based on the reaction of bacterial cell wall on staining solutions

A

T

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5
Q

Two Types of Smear

prepared from colonies growing from plated media

A

Indirect Smear

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6
Q

Two Types of Smear

prepared from clinical specimen

A

Direct Smear

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7
Q

Staining Procedure Gram Stain
Stain :
Mordant :
Decolorizer :
Counterstain :

A

Stain : Crystal Violet for 1 minute
Mordant : Gram’s Iodine for 1 minute
Decolorizer : Acetone-alcohol or 95% Ethanol for 10 seconds
10 seconds
Counterstain : Safranin O for 30 seconds

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8
Q

T/F In Gram staining smear should have pink/red or clear background and cells are overlapping

A

F, cells should not be overlapping

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9
Q

Gram staining

Examine smear _____/____ scanning

A

Horizontal / Vertical Scanning

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10
Q

Microscopic Examination : Direct Staining

Microscopic Structures Present

A

Epithelial Cells, PMNs, RBCs, Host cellular material

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11
Q

Gram Staining

Microscopic Examination : Enumeration of Cells under LPO

1+

A

1+ = </LPF

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12
Q

Gram Staining

Microscopic Examination : Enumeration of Cells under LPO

2+

A

2+ = 1-9/LPF

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13
Q

Gram Staining

Microscopic Examination : Enumeration of Cells under LPO

3+

A

3+ = 10-25/LPF

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14
Q

Gram Staining

Microscopic Examination : Enumeration of Cells under LPO

4+

A

4+ = >25/LPF

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15
Q

Gram Staining

Microscopic Examination : Enumeration of Bacteria under OIO

1+

A

1+ (rare or occasional) = <1/OIF

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16
Q

Gram Staining

Microscopic Examination : Enumeration of Bacteria under OIO

2+

A

2+ (few) = 1-5/OIF

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17
Q

Gram Staining

Microscopic Examination : Enumeration of Bacteria under OIO

3+

A

3+ (moderate) = 6-30/OIF

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18
Q

Gram Staining

Microscopic Examination : Enumeration of Bacteria under OIO

4+

A

4+ (heavy) = >30/OIF

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19
Q

Differential Staining

Principle : For the detection of Tubercle bacilli in sputum specimen

A

AFS

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20
Q

AFS

What does it detect in sputum specimens

A

Tubercle bacilli

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21
Q

T/F In AFS, Tubercle bacili resist decolorization w/ acid-alcohol and appear red on a blue / green background after decolorization

A

T

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22
Q

AFS

Causative agent of Mycobacterium Tuberculosis

A

Tubercle bacilli

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23
Q

AFS

Primary diagnostic and monitoring tool for TB

A

Direct Sputum Smear Microscopy

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24
Q

AFS

Specimen Considerations
Time

A

Collected preferably early morning

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25
Q

AFS

Sputum should be ____

A

Sputum should be thick, cloudy, and sticky

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26
Q

AFS Procedure

Smear should be spread properly showing ____ patterns

A

concentric / coil-like patterns

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27
Q

AFS

Stain

A

0.3% Ziehl’s Carbol Fuchsin

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28
Q

AFS

Decolorizer

A

Acid alcohol

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29
Q

AFS

Counterstain

A

0.3% Methylene blue / Malachite Green

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30
Q

AFS Microscopic Examination

Manner of scanning

A

From left to right go downward a bit and then right to left

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31
Q

AFS

Microscopic Structures Present

A

Squamous Cells, Leukocytes, Mucus Threads, Bacterial cells aside from AFB

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32
Q

AFS

Color of Bacterial Cells aside from AFB

A

Blue and green

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33
Q

AFS

Color & Morphology of AFB

A

red or pink bacilli; thin,
filamentous or beaded

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34
Q

Enumeration of AFB under OIO

+n

A

+n : 1 - 9 AFB seen in 100 OIF

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35
Q

Enumeration of AFB under OIO

1+

A

1+ : 10-99 AFB seen in 100 OIF

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36
Q

Enumeration of AFB under OIO

2+

A

2+ : 1-10 AFB/OIF in at least 50
fields

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37
Q

Enumeration of AFB under OIO

3+

A

3+ : >10 AFB/OIF in at least 20
fields

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38
Q

Special Staining

T/F Pathogenic organisms have a protected covering made up of polysaccharide polymer and polypeptide

A

T

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39
Q

Special Staining

Stain used for Capsules

A

India ink / Nigrosin

40
Q

Special Staining

Appears halo-like against stained
amorphous background

A

Capsule

41
Q

Special Staining

A virulence factor which inhibits phagocytosis

A

Capsule

42
Q

Special Staining : Capsule

Specimen

A

Klebsiella pneumoniae on a nutrient agar slant or plate

43
Q

Special Staining

multilayered ovoid or spherical structures present inside Gr (+) bacteria

A

Endospores

44
Q

Special Staining

Seen among Bacillus and Clostridium

A

Endospore

45
Q

Special Staining

Organisms with endospores

A

Bacillus & Clostridium

46
Q

Special Staining

Can be found centrally, subterminal, or terminal

A

Endospore

47
Q

Special Staining : Spore

T/F Endospores are highly resistant to heat, desiccation, and chemical disinfectant

A

T

48
Q

Special Staining : Spore

Main component of spore

A

Calcium dipicolinate or dipocolonic acid calcium complex

49
Q

Special Staining : Spore

appear as _____ structure

A

clear structure

50
Q

Special Staining : Spore

Technique used

A

Schaeffer-Fulton Method

51
Q

Special Staining : Spore

Specimen

A

Pure colonies of Bacillus subtilis in a nutrient slant or plate

52
Q

Special Staining : Spore

Stain used

A

6.5% Malachite green

53
Q

Special Staining : Spore

Counterstain + time

A

Safranin O for 30 seconds

54
Q

Special Staining : Spore

Endospores stain ____

A

green

55
Q

Special Staining : Spore

Vegetative cells stain ____

A

red/pink

56
Q

Special Staining

stained using Leifson staining technique

A

Flagella

57
Q

Special Staining : Flagellar

most common stain used

A

Leifson staining method

58
Q

Special Staining : Flagellar

Leifson staining technique
Primary Stain

A

Basic fuchsin

59
Q

Special Staining : Flagellar

Leifson staining technique
Mordant

A

Tannic Acid

60
Q

Special Staining : Flagellar

Leifson staining technique
Counterstain

A

Methylene Blue

61
Q

Special Staining : Flagellar

Properly prepared bacterial suspension = [appearance]

A

slightly cloudy

62
Q

Special Staining : Flagellar

when do you wash the smear with distilled water [2 indicators]

A
  1. golden precipitate is formed on the dye surface
  2. precipitates appear on the smear
63
Q

Special Staining : Flagellar

color of bacterial cells and flagella

A

red

64
Q

Special Staining : Flagellar

devoid of flagella

A

Atrichous

65
Q

Special Staining : Flagellar

flagella on one pole/end

A

Monotrichous

66
Q

Special Staining : Flagellar

a tuft/group of flagella on one pole/end

A

Lophotrichous

67
Q

Special Staining : Flagellar

flagella on both poles/ends

A

Amphitrichous

68
Q

Special Staining : Flagellar

surrounded with flagella

A

Peritrichous

69
Q

Test for Motility

non-staining method for demonstration of presence of flagella

A

Hanging Drop Method

70
Q

Test for Motility

long, proteinaceous filaments for
locomotion

A

Flagella

71
Q

Test for Motility

Considered as a virulence factor

A

Flagella

72
Q

Test for Motility

Presence or absence is interpreted as

A

Presence = motile
absence = non-motile

73
Q

Test for Motility

used for serotyping

A

Flagellar antigens

74
Q

Test for Motility : HDM

Appearance of bacterial cells

A

appear translucent with slight greenish color

75
Q

Test for Motility : Hanging Drop Method

Specimen

A

Broth cultures of known motile or non-motile bacteria

76
Q

Test for Motility : HDM

True Motility : [movement]

A

bacterial cells exhibit directional
purposeful movement

77
Q

Test for Motility : HDM

Non-motile bacteria movement

A

Brownian movement

78
Q

Test for Motility : HDM

random jiggling or shaking due to bombardment of water molecules

A

Brownian movement

79
Q

Test for Motility : Use of SIM

Specimen

A

Broth cultures of known motile or non-motile bacteria

80
Q

Test for Motility : Use of SIM

Medium

A

Sulfide Indole Medium

81
Q

Test for Motility : Use of SIM

Incubation of medium
____ hrs at ____ C

A

18 - 24 hrs at 35 - 37°C

82
Q

Test for Motility : Use of SIM

Growth Pattern : bacterial growth is away from the site of inoculation

A

Motile

83
Q

Test for Motility : Use of SIM

Growth Pattern : bacterial growth is limited to the site of inoculum

A

Non-motile

84
Q

Test for Motility : Use of SIM

Appearance of Motile

A

Appears diffuse or hazy

85
Q

Test for Motility : Use of SIM

Appearance of Non-motile

A

Surrounding medium is clear

86
Q

Test for Motility : Use of SIM

Inoculate culture by making a [action] down the center of the tube about [depth] of the medium

A

stab using inoculating needle;
3/4 the depth

87
Q

Gram Staining Procedure

Reagents + Time

A
  1. Crystal Violet 1 minute
  2. Gram’s Iodine 1 minute
  3. 95% Ethanol or Acetone Alcohol 10 seconds
  4. Safranin O or Carbol Fuchsin 30 seconds
88
Q

AFS : Ziehl Neelsen Staining Procedure

Reagents + Time

A
  1. Carbol Fuchsin 10 mins
  2. Steam
  3. Acid Alcohol 3 mins
  4. MB / MG less than 1 minute
89
Q

AFS

Reason why we use heat after flooding Carbol Fuchsin

A

to enhance penetration of dye to the cells

90
Q

AFS

Reason for using filter paper during staining

A

to avoid drying out

91
Q

Test for Motility HDM

Petroleum jelly is used to

A

seal cover slip & drop of bacteria; to keep it from drying out

92
Q

Spore Staining Procedure

Reagents + Time

A
  1. Malachite Green 10 mins
  2. Safranin O 30 seconds
93
Q

Flagellar Staining Procedure

Reagents + Time

A
  1. Leifson dye 7-15 mins
94
Q

Test for Motility HDM

Slide used

A

concavity slide

95
Q

Test for Motility HDM

Bacterial cells would appear as

A

translucent with slight
greenish color

[3Y1S] MT 6320 (8 decks)

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