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BACTERIOLOGY - LAB > Culture > Flashcards

Culture Flashcards

1
Q

PRINCIPLES OF BACTERIAL CULTIVATION

A

✓Grow and isolate all bacteria present in a
clinical specimen
✓Determine which of the bacteria that grow
are most likely causing infection and which
are likely contaminants
✓Obtain sufficient growth of clinically relevant
bacteria to allow identification,
characterization, and susceptibility testing

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2
Q

process of growing microorganisms in culture by taking bacteria
from the infection site (in vivo environment) by some means of
specimen collection and growing them in the artificial
environment of the laboratory (in vitro environment)

A

Cultivation

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3
Q

→nutrient material prepared for the growth of microorganisms in a
laboratory

A

Culture Medium

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4
Q

composed of mixture of nutrients: Carbon, Nitrogen, Sulfur,
Phosphorus, Hydrogen, Oxygen and Buffer

A

Culture Medium

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5
Q

Inhibitory agents→ facilitate isolation of desired organism while
suppressing the growth of other organism

A

Culture Medium

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6
Q

microbes that are introduced into a culture medium to initiate
growth

A

Inoculum

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7
Q

microbes that grow and multiply in or on a culture
medium

A

CULTURE

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8
Q

Types of CULTURE

A

Pure (Axenic) Culture
Mixed Culture
Stock Culture

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9
Q

Composed of only one species

A

Pure (Axenic) Culture

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10
Q

Composed of more than one species

A

Mixed Culture

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11
Q

Composed of several species contained in a separate culture medium—one specie per culture medium

A

Stock Culture

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12
Q

Grown in a large volume of broth and then divided into small freezer vials—lengthen the shelf life of specimen to at least a year

A

Stock Culture

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13
Q

Sulfated polymer made up of D-galactose, 3,6-anhydro-Lgalactose, and D-glucoronic acid and usually derived from red
algae

A

Agar

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14
Q

melt at 80°C-90°C (100°C) and solidify at 40°C-50°C

A

Agar

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15
Q

cooling temperature for distribution of culture
medium into Petri plates

A

55°C-60°C

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16
Q

Amount of molten agar transferred to a sterile
plates

A

20-25 ml

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17
Q

CLASSIFICATION OF CULTURE MEDIA

A

I. According to Physical State or Consistency
II. According to Composition
III. According to the Dispensing or Distribution
Method for the Medium
IV. According to Use

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18
Q

(CULTURE MEDIA)
According to Physical State or Consistency

A

Liquid Medium
Semi-solid Medium
Solid Medium

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19
Q

does not contain any amount of agar or solidifying
substances

A

Liquid Medium

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20
Q

allows growth of aerobes, anaerobes and facultative
anaerobes

A

Liquid Medium

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21
Q

Example of Liquid Medium

A

✓Nutrient Broth
✓Brain Heart Infusion (BHI)
✓Trypticase Soy Broth (TSB)
✓Thioglycollate (THIO)

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22
Q

contains 0.5% to 1% agar

A

Semi-solid Medium

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23
Q

observed bacterial motility and detect indole and
sulfide production

A

Semi-solid Medium

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24
Q

Example of Semi-Solid Medium

A

Sulfide Indole Motility (SIM) Medium

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25
Q

contains 2% to 3% agar

A

Solid Medium

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26
Q

(CULTURE MEDIA)
According to Composition

A

Synthetic or Defined Medium
Non-synthetic or Complex Medium
Tissue Culture Medium

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27
Q

exact chemical composition of the ingredients
is known (commercially prepared culture media)

A

Synthetic or Defined Medium

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28
Q

used for research purposes as either a liquid or
solid medium

A

Synthetic or Defined Medium

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29
Q

preferred for the isolation of cyanobacteria and
chemoorganotrophs

A

Synthetic or Defined Medium

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30
Q

Example of Synthetic or Defined Medium

A

BG-11 medium

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31
Q

precise composition of some or all of the nutritive
substances used is not known (Peptone, Meat and Yeast Extracts)

A

Non-synthetic or Complex Medium

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32
Q

useful for the isolation of medically significant bacteria

A

Non-synthetic or Complex Medium

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33
Q

Example of Non-synthetic or Complex Medium

A

Nutrient Broth (NB) Medium, TSB and MAC Agar

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34
Q

contains living tissues

A

Tissue Culture Medium

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35
Q

used for obligate intracellular bacteria—Rickettsia and Chlamydia

A

Tissue Culture Medium

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36
Q

Example of Tissue Culture Medium

A

HeLa 229→ human cervical tissue, Chlamydia
McCoy and W 138→ fibroblasts, Chlamydia
Embryonated Egg→ propagation of Rickettsia

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37
Q

(CULTURE MEDIA)
According to the Dispensing or Distribution Method for the Medium

A

Plated Media
Tubed Media

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38
Q

distributed into sterile petri dish

A

Plated Media

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39
Q

distributed in sterile test tube

A

Tubed Media

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40
Q

Types of TUBED MEDIA

A
  1. Slant
  2. Butt-Slant
  3. But
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41
Q

Example of Tubed Media

A

TSI, SIM, Simmon’s Citrate Agar (SCA),
Lysine Iron Agar (LIA)

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42
Q

(CULTURE MEDIA)
According to Use

A

1) Simple Media, Supportive Media or General Purpose Media
2) Enrichment Media
3) Enriched Media
4) Differential Media
5) Selective Media

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43
Q

routinely used in the laboratory and without additional supplement

A

Simple Media, Supportive Media or General Purpose Media

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44
Q

support growth of most non-fastidious bacteria to grow at natural rates, without providing advantage to any particular bacteria

A

Simple Media, Supportive Media or General Purpose Media

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45
Q

usually composed of meat and soybean extracts

A

Simple Media, Supportive Media or General Purpose Media

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46
Q

Example if Simple Media, Supportive Media or General Purpose
Media

A

✓Nutrient Agar
✓Nutrient Broth
✓TSB

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47
Q

enhance the growth of particular organisms (pathogens) and
suppress the growth of normal flora present in specimen

A

Enrichment Media

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48
Q

contain specific nutrients and without additional supplements

A

Enrichment Media

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49
Q

incubated for a certain period and then subcultured to
isolate the desired organism

A

Enrichment Media

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50
Q

can also be used as a supplement to agar plates to detect
aerobes, anaerobes and microaerophlies (THIOGLYCOLLATE)

A

Enrichment Media

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51
Q

Example of Enrichment Media

A

✓Alkaline Peptone Water (APW)
✓Selenite F
✓Thioglycollate
✓Tetrathionate
✓Gram-Negative Broth
✓Lim Broth (Todd Hewitt with CNA)

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52
Q

→promote growth of Vibrio spp. before inoculation
into Thiosulfate-Citrate-Bile-Salts(TCBS) Agar
→adjusted to pH 8.5

A

Alkaline Peptone Water (APW)

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53
Q

isolation of Salmonella from feces, urine and water sample

A

Selenite F

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54
Q

general support enrichment medium that promotes the
growth of almost all non-fastidious bacteria

A

Thioglycollate

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55
Q

Components of Thioglycollate:

A

▪ Dextrose, Vitamin K1, and Hemin have been used to
modify the basic thioglycollate formula
▪ 0.075% agar
▪ Resazurin→ oxidation-reduction indicator
▪ Thioglycolic Acid→ reducing agent

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56
Q

-Selective enrichment broth for the isolation of
Salmonella and Proteus
-Bile Salt and Thiosulfate→ suppresses the growth of
other coliform bacilli

A

Tetrathionate

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57
Q

Bile Salt and Thiosulfate

A

suppresses the growth of
other coliform bacilli

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58
Q

solation of Salmonella and Shigella

A

Gram-Negative Broth

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59
Q

Enrichment and Selective medium

A

Gram-Negative Broth

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60
Q

→ inhibit gram-positive organisms

A

Sodium Citrate and Sodium Desoxycholate (a
bile salt)

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61
Q

→ primary carbon source

A

Mannitol

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62
Q

Gram-Negative Broth

A

Sodium Citrate and Sodium Desoxycholate (a
bile salt)

Mannitol

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63
Q

Group B Streptococci

A

Lim Broth (Todd Hewitt with CNA)

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64
Q

→media with additional supplements necessary
for growth of fastidious organisms

A

Enriched Media

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65
Q

Supplements: Blood, Vitamins, Serum, Peptone
and Yeast Extract

A

Enriched Media

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66
Q

solid type media

A

Enriched Media

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67
Q

Example of Enriched Media

A

✓Blood Agar Plate (BAP)

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68
Q

contains 5% defibrinated blood

A

Blood Agar Plate (BAP)

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69
Q

Differentiate haemolytic pattern of
bacteria

A

Blood Agar Plate (BAP)

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70
Q

Choices of blood in Blood Agar Plate

A

Sheep, Horse, Rabbit

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71
Q

→blood has been chemically-treated or heat-treated
(80°C) to lyse the RBC

A

✓Chocolate Agar Plate

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72
Q

isolation of fastidious microorganisms:
***Neisseria gonorrhoeae and Haemophilus spp.

A

✓Chocolate Agar Plate

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73
Q

(Chocolate Agar Plate)
isolation of fastidious microorganisms:
_______

A

Neisseria gonorrhoeae and Haemophilus spp.

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74
Q

“X” factor

A

Hemin

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75
Q

“V” factor

A

Nicotinamide Adenine Dinucleotide (NAD)

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76
Q

allow the visualization of metabolic differences
between groups of bacteria

A

Differential Media

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77
Q

distinguishes organisms growing together by their
diffrences in cultural characeristics

A

Differential Media

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78
Q

allow grouping of microbes based on different
characteristics demonstrated on the medium

A

Differential Media

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79
Q

Example of Differential Media

A

BAP
Eosin Methylene Blue (EMB)
Hektoen Enteric Agar (HEA)

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80
Q

→Lactose and Sucrose
→Eosin and Methylene Blue

A

Eosin Methylene Blue (EMB)

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81
Q

→differentiate Lactose Fermenter (pink colonies)
from Non-Lactose Fermenter (colorless colonies)

A

MacConkey Agar

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82
Q

Components of MacConkey Agar

A

▪ Lactose
▪ Bile Salts
▪ Crystal Violet→ inhibit gram-positive bacteria and
fungi
▪ Neutral Red→ pH indicator

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83
Q

(MacConkey Medium)
pH indicator

A

Neutral Red

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84
Q

(MacConkey Agar)
inhibit gram-positive bacteria and
fungi

A

Crystal Violet

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85
Q

support the growth of one type or group of microbes but not
another

A

Selective Media

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86
Q

contain inhibitory substances such as antimicrobials, dyes, or
alcohol which inhibit the growth of other organisms while
promoting the growth of the desired organism

A

Selective Media

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87
Q

INHIBITORY AGENTS

A

1) Inhibit growth of Gram-Positive Microorganism
2)Inhibit growth of Gram-Negative Microorganisms
3) Prevent Swarming of Proteus

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88
Q

Inhibit growth of Gram-Positive Microorganism

A

✓Crystal or Gentian Violet
✓Basic or Carbol Fuchsin
✓Bile Salts
✓Sodium Desoxycholate

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89
Q

Inhibit growth of Gram-Negative Microorganisms

A

✓Potassium Tellurite
✓Sodium Azide

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90
Q

Prevent Swarming of Proteus

A

✓Alcohol
✓Chloral Hydrate

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91
Q

Examples of Selective Media

A

✓HEA
✓MAC
✓Xylose Lysine Deoxycholate (XLD)
✓Bismuth Sulfite Agar (BSA)
✓Mannitol Salt Agar (MSA)
✓Thayer Martin Agar (TMA)
✓Salmonella-Shigella Agar (SSA)
✓TCBS

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92
Q

→ Salmonella spp. and Shigella spp.
→bile salts and dyes (bromthymol
blue and acid fuchsin)

A

Hektoen Enteric (HE) Agar

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93
Q

Hektoen Enteric (HE) Agar
pH indicator

A

Bromthymol Blue

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94
Q

(Hektoen Enteric (HE) Agar)
→ H2S indicator

Salmonella→ black precipitate

A

Ferric Ammonium Citrate

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95
Q

Shigella spp. and Salmonella spp.

A

Xylose-Lysine-Desoxycholate (XLD) Agar

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96
Q

Components Xylose-Lysine-Desoxycholate (XLD) Agar

A

▪Lysine, Lactose, Xylose and Sucrose
▪0.25% Sodium Desoxycholate
→ inhibits gram-positive
bacteria
▪Phenol Red→ pH indicator
Ferric Ammonium Citrate→ H2S indicator

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97
Q

(Xylose-Lysine-Desoxycholate (XLD) Agar)

inhibits gram-positive
bacteria

A

▪0.25% Sodium Desoxycholate

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98
Q

(Xylose-Lysine-Desoxycholate (XLD) Agar)

pH indicator

A

Phenol Red

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99
Q

(Xylose-Lysine-Desoxycholate (XLD) Agar)

H2S indicator

A

Ferric Ammonium Citrate

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100
Q

colonies are red with
black center

A

Salmonella

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101
Q

MEDIA FOR GRAM-POSITIVE BACTERIA

A

1) Columbia CNA with Blood
2) Phenylethyl Alcohol (PEA) Agar

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102
Q

three peptone sources and 5% defibrinated
sheep blood

A

Columbia CNA with Blood

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103
Q

suppress the growth of most gram-negative
organisms

A

Colistin (C) and Nalidixic Acid (NA)

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104
Q

sheep blood agar supplemented with
phenylethyl alcohol to inhibit the growth of
gram-negative bacteria

A

Phenylethyl Alcohol (PEA) Agar

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105
Q

Culture Media for Neisseria spp.

A

1) THAYER-MARTIN
2) MODIFIED THAYER-MARTIN AGAR
3) TRANGROW MEDIUM
4) MARTIN-LEWIS AGAR
5) NEW YORK CITY MEDIUM

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106
Q

enriched Chocolate Agar with supplement B or
Isovitale X

A

THAYER-MARTIN

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107
Q

Antibiotic Components:

A

▪ Colistin
▪ Vancomycin
▪ Nystatin

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108
Q

(MODIFIED THAYER-MARTIN AGAR & THAYER-MARTIN)
Antibiotic Component that
inhibit gram-negative bacteria

A

Colistin

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109
Q

(MODIFIED THAYER-MARTIN AGAR & THAYER-MARTIN)
Antibiotic Component that inhibit gram-positive bacteria

A

Vancomycin

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110
Q

(MODIFIED THAYER-MARTIN AGAR & THAYER-MARTIN)
Antibiotic Component that inhibit yeast

A

Nystatin

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111
Q

→Neisseria gonorrhoeae and Neisseria meningitides
→chocolatized blood + antibiotics

A

MODIFIED THAYER-MARTIN AGAR

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112
Q

Antibiotic components

A

▪ Colistin
▪ Vancomycin
▪ Nystatin
▪ Trimethoprim Lactate

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113
Q

(MODIFIED THAYER-MARTIN AGAR)
Antibiotic component that inhibit Proteus spp

A

Trimethoprim Lactate

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114
Q

Thayer-Martin with glucose, 2% agar,
Trimethoprim Lactate and CO2
incorporated in bottle

A

TRANGROW MEDIUM

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115
Q

substitute Anisomycin*** for Nystatin and
higher concentration of vancomycin

A

MARTIN-LEWIS AGAR

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116
Q

Modified Thayer-Martin with substitution of
Amphotericin B*** for Nystatin

A

NEW YORK CITY MEDIUM

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117
Q

OTHER SELECTIVE MEDIA

A

Gentamicin Blood Agar
Bacitracin Chocolate Agar
Blood Agar Plate with Ampicillin

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118
Q

Gentamicin Blood Agar

A

Streptococcus

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119
Q

Bacitracin Chocolate Agar

A

Haemophilus

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120
Q

Blood Agar Plate with Ampicillin

A

Aeromonas

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121
Q

used in the primary isolation of enteric Gram-Negative bacteria

A

Selective and Differential Media

122
Q

Example of Selective and Differential Media

A

✓Endo Agar
✓XLD Agar
✓MAC Agar
✓EMB Agar

123
Q

solate bacteria with specific growth requirements

A

Special Media

124
Q

specially prepared to support the growth of specific
microorganisms

A

Special Media

125
Q

Examples of Special Media

A

✓Middlebrook 7H-10 Agar
✓Fletcher Medium
✓“W”or Winsconsin Medium
✓Bordet-Gengou Agar
✓Thayer Martin
✓MacBride
✓Dieudonn’s Medium

126
Q

Middlebrook 7H-10 Agar

A

M. tuberculosis

127
Q

Fletcher Medium

A

Leptospira

128
Q

“W”or Winsconsin Medium

A

Brucella

129
Q

Bordet-Gengou Agar

A

Bordetella pertussis

130
Q

Thayer Martin

A

Nesseria

131
Q

Dieudonn’s Medium

A

Vibrio cholerae

131
Q

MacBride

A

Listeria monocytogenes

132
Q

protein rich medium composed of whole
egg and malachite green and supports
the growth of Mycobacteria

A

Lowenstein-Jensen

133
Q

Sterilization: Inspissation not Autoclaving

A

Lowenstein-Jensen

134
Q

selective for the isolation of Vibrio

A

Thiosulfate-Citrate-Bile Salts-Sucrose (TCBS)
Agar

135
Q

“Special Medium”

A

Thiosulfate-Citrate-Bile Salts-Sucrose (TCBS)
Agar

136
Q

Sterilization: Boiling not Autoclaving

A

Thiosulfate-Citrate-Bile Salts-Sucrose (TCBS)
Agar

137
Q

General Steps in Preparation of Culture Medium

A

TUBE METHOD
PLATED MEDIUM
INOCULATION OF MEDIA

138
Q

TUBE METHOD

A
  1. Weighing
  2. Dissolving
  3. Titration
  4. Distribution
  5. Sterilization
139
Q

→ weigh the different ingredients then
place in clean, dry containers

A

Weighing

140
Q

→ add the exact amount of solvent to
the ingredients and then dissolve by
heating

A

Dissolving

141
Q

→ adjustment to the right pH : 7.2-7.4

A

Titration

142
Q

→ distribute in test tubes

A

Distribution

143
Q

PLATED MEDIUM

A

1.Weighing
2.Dissolving
3.Titration
4.Sterilization
5.Distribution

144
Q

INOCULATION OF MEDIA
SPECIMEN CONSIDERATIONS

A
  1. Sterile body fluids, pus, urine and sputum
  2. Specimens received on swabs
  3. Specimens that require direct or “bedside”
    inoculations
145
Q

Inoculated directly into the selected media

A

Sterile body fluids, pus, urine and sputum

146
Q

Can be inoculated directly into the culture
media

A

Specimens received on swabs

147
Q

Specimens that require direct or “bedside”
inoculations

A

Blood, Genital specimens, Corneal
scrapings, Sterile fluids like Synovial and
Peritoneal fluids and Nasopharyngeal Swabs for
isolation of Bordetella pertussis

148
Q

most common manner of inoculation

A

Streaking

149
Q

Placement of fluid specimen or swabs into a broth or
liquid culture media

A

Streaking

150
Q

✓Stabbing of medium is usually performed with group A _______________ to create anaerobiosis and promote sub-surface hemolysis

A

streptococci

151
Q

inoculation technique used for antimicrobial sensitivity test

A

Overlapping inoculation

152
Q

Inoculation of Tubed Media

A

a. Liquid Medium
b. Slant Medium
c. Butt Medium
d. Butt/Slant Medium

153
Q

Liquid medium is inoculated with the use of

A

Sterile Pasteur Pipet

154
Q

➢inoculate by shaking a previously heated wire loop or needle

A

Liquid Medium

155
Q

➢With the use of a wire loop or needle, transfer the inoculum to the bottom of the slant and streak in a zig-zag manner across the entire surface toward the mouth of the tube

A

Slant Medium

156
Q

➢Just stab the medium with an inoculating
needle

A

Butt Medium

157
Q

➢Inoculate the butt first by stabbing the
needle to the bottom of the medium and
then streak the surface in a zig-zag manner
toward the mouth of the tube

A

Butt/Slant Medium

158
Q

Inoculation of Plated Media

A

a. Streak Plate Technique
b. Pour Plate
c. Streak-Pour Plate

159
Q

inoculate by isolating organisms in pure culture

A

Streak Plate Technique

160
Q

Inoculation that determine the approximate number of viable organisms in a liquid such as water, milk, urine or broth culture

A

Pour Plate

161
Q

Inoculation for studying hemolysis

A

Streak-Pour Plate

162
Q

→calibrated loop inserted into the
urine and transferred to the culture
medium by making a single streak
down the center of the plate
→without flaming, the loop is
streaked back and forth through
the original inoculum

A

Quantitative Isolation

163
Q

What is the specimen used for Quantitative Isolation?

A

Urine Specimens

164
Q

Quantitative Isolation use a calibrated loop to deliver a specified volume of

A

0.01 or 0.001 mL

165
Q

Manner of Reporting (Grading) of Growth on Plate

4+

A

many, heavy growth; growth is up to
the fourth quadrant

166
Q

Manner of Reporting (Grading) of Growth on Plate

3+

A

moderate growth; growth is up to
the third quadrant

167
Q

Manner of Reporting (Grading) of Growth on Plate

2+

A

few or light growth; growth is in the
second quadrant

168
Q

Manner of Reporting (Grading) of Growth on Plate

1+

A

rare growth; growth is in the first
quadrant only

169
Q

METHODS OF OBTAINING PURE CULTURE

A

✓Streak-plate method
✓Pour- plate method
✓Use of selective media and media
containing antibiotic
✓Animal inoculation test

170
Q

→usually measured in millimeters or described as pinpoint, small, medium, large

A

Colony Size

171
Q

→includes form, elevation, and margin of the colony

A

Colony Shape

172
Q

→glistening, opaque, dull, dry, transparent

A

Colony surface appearance

173
Q

→hemolytic pattern on blood agar, changes in color of pH indicators, pitting of the agar surface

A

Changes in agar media resulting from bacterial growth

174
Q

→certain bacteria produce distinct odors that can be helpful in preliminary identification

A

Odor

175
Q

COLONY MORPHOLOGY

A

✓Colony size
✓Colony pigmentation
✓Colony shape
✓Colony surface appearance
✓Changes in agar media resulting from bacterial growth
✓Odor

176
Q

Types of Colony

A

A. Mucoid (M) Colony
B. Smooth (S) Colony
C. Rough (R) Colony

177
Q

(Colony)
→exhibits a water-like, glistening, confluent appearance
→characteristic of organisms that form slimes or welldeveloped capsule

A

Mucoid Colony

178
Q

(Colony)
→uniform texture and homogeneity
→asily emulsified in NSS
→haracteristic of freshly isolated wild type
microorganisms (virulent microorganisms)

A

Smooth Colony

179
Q

Examples of Bacteria in M Colony

A

K. pneumoniae, S. pneumoniae, H. influenzae

180
Q

Examples of Bacteria in S Colony

A

Salmonella, Shigella, E. coli, Serratia, Proteus

181
Q

(Colony)
→granulated and rough in appearance
→hard to emulsify in NSS
→usually produced by mutant strains that lack
surface proteins or polysaccharides indicating loss of virulence

A

Rough (R) Colony

182
Q

Examples of Bacteria in R Colony

A

Rough forms of enteric bacteria

183
Q

Exception for Rough Colony

A

R forms of B. anthracis and human and
bovine types of M. tuberculosis (more
virulent)

184
Q

(Colony Characteristic)
→observed in the media immediately surrounding or underneath the colony is a reaction caused by enzymatic or toxin activity of bacteria
→presumptive identification of streptococci and enterococci

A

Hemolysis

185
Q

→partial lysing of erythrocytes in a BAP around and under the colony that results in a green discoloration of the medium

A

α-Hemolysis

186
Q

Examples of Bactria for α-Hemolysis

A

Streptococcus pneumoniae and certain
viridans streptococci

187
Q

Culture Media for Hemolysis

A

Blood Agar Plate

188
Q

→complete clearing of erythrocytes in BAP around or under the colonies because of the complete lysis of RBCs

A

β-Hemolysis

189
Q

wide, deep, clear zone
of β-hemolysis,

A

Streptococcus pyogenes

190
Q

→narrow, diffuse zone of β-hemolysis close to the colony

A

Streptococcus agalactiae and Listeria
monocytogenes

191
Q

(Colony Characteristic)
→ described as large, medium, small, or pinpoint
→gram-positive bacteria produce smaller
colonies than gram-negative bacteria

A

Size

192
Q

In BAP, small, white colonies are

A

gram-positive cocci

193
Q

In BAP, large, gray, mucoid colonies are

A

enteric gram-negative rods

194
Q

(Colony Characteristic)
→edge of the colonies
→described as smooth, filamentous, rough or rhizoid, or irregular

A

Form of Margin

195
Q

“Medusa Heads”—filamentous appearance

A

Bacillus anthracis

196
Q

swarming phenomena in what bacteria

A

Proteus mirabilis and Proteus vulgaris

197
Q

hazy blanket of growth on the
surface that extends well beyond
the streak lines

A

Swarming

198
Q

bacteria with rough edges

A

Diphtheroids

199
Q

(Colony Characteristics)
→determined by tilting the culture
plate and looking at the side of the
colony
→raised, convex, flat, umbilicate
(depressed center, concave—an
“innie”), or umbonate (raised or
bulging center, convex—an “outie”)

A

Elevation

200
Q

umbilicate colonies (unless the colonies
are mucoid)

A

S. pneumoniae

201
Q

→ convex colonies β-hemolytic streptococci
→ flat colonies

A

S. aureus

202
Q

(Colony Characteristics)
→transparent, translucent, or opaque

A

Density

203
Q

Density of β-Hemolytic streptococci except group B (S. agalactiae)

A

→ translucent

204
Q

Density of S. agalactiae

A

semiopaque

205
Q

Density of Staphylococci and other gram-positive bacteria

A

Opaque

206
Q

Density of most gram-negative rods

A

opaque

207
Q

Density of Bordetella pertussis

A

→ shiny, similar to a half-pearl,
on blood-containing media

208
Q

Color of Coagulase-negative staphylococci

A

white

209
Q

Color of Enterococcus spp.

A

gray

210
Q

Color of Certain Micrococcus spp. and Neisseria (nonpathogenic) spp.

A

yellow or off-white

211
Q

Color of Diphtheroids

A

buff

212
Q

Color of most gram-negative rods

A

gray on BAP

213
Q

(Colony Characteristic)
→determined by touching the colony with a sterile loop
→ brittle (splinters), creamy (butyrous), dry, or waxy

A

Consistency

214
Q

Consistency of S, aureus

A

Creamy

215
Q

COnsistency of Nocardia spp.

A

brittle, crumbly, and wrinkled, resembling bread crumbs on a plate

216
Q

Consistency of diphtheroid

A

dry and waxy

217
Q

Pseudomonas aeruginosa pigment

A

green

218
Q

Serratia marcescens pigment

A

brick-red (especially at room temperature)

219
Q

Chromobacterium violaceum pigment

A

purple

220
Q

Prevotella meninogenica pigment

A

brown-black (anaerobic)

221
Q

Kluyvera spp. pigment

A

blue

222
Q

Odor of S. aureus

A

old sock—Mannitol Salt Agar

223
Q

Odor of P. aeruginosa

A

fruity or gapelike

224
Q

Odor of P. mirabilis

A

putrid

225
Q

Odor of Haemophilus spp.

A

musty basement, “mousy” or
“mouse nest” smell

226
Q

Nocardia spp. odor

A

freshly plowed field

227
Q

OTHER MEDIA for Staphylococcus aureus

A

✓CNA
✓Chapman stone agar
✓Vogel-Johnson medium
✓PEA, MSA

228
Q

OTHER MEDIA for Streptococci

A

✓ PEA, Todd-Hewitt broth, CAN

229
Q

OTHER MEDIA for Neisseria

A

✓ Thayer-Martin
✓ Modified Thayer-Martin
✓ Transgrow medium
✓ Martin-Lewis
✓ New York City medium

230
Q

OTHER MEDIA for Mycobacterium

A

✓ Dubos oleic acid medium
✓ Middlebrook 7H10 or 7H11
✓ Mitchison’s selective 7H11
✓ Petragnani
✓ LJ
✓ Dorset egg
✓ American Thoracic Society
medium
✓ Bactec 12B medium
✓ Middlebrook 7H9 broth

231
Q

OTHER MEDIA for Corynebacterium diphthriae

A

✓Loeffler’s coagulated serum media
✓Modified tinsdale
✓Cystine tellurite media
✓Pai’s coagulated egg media

232
Q

OTHER MEDIA for Listeria monocytogenes

A

✓ McBride agar
✓ PEA
✓ Cold enrichment technique

233
Q

OTHER MEDIA for Erysipelothrix rhusiopathiae

A

✓ BAP, tellurite, gelatin (testtube brush growth)
✓ BHIA w/ 1% glucose, 5% CO2
at 35-37C

234
Q

OTHER MEDIA for Nocardia

A

✓ LJ, BHIA, SDA

235
Q

OTHER MEDIA for C. perfringens

A

✓BAP = double zone hemolysis
✓Chopped meat glucose media = +gas
✓Milk media = stormy ferm.

236
Q

OTHER MEDIA for C. tetani

A

BAP = swarming, faint beta hemolysis

237
Q

OTHER MEDIA for C. botulinum

A

BAP = beta hemolysis

238
Q

OTHER MEDIA for C. difficile

A

BAP = yellow green w/ horse stable odor
CCFA = yellow ground glass colonies

239
Q

OTHER MEDIA for Actinomyces israelii

A

BHIA = molar tooth colony/ breadcrumb-like, raspberry or smooth colony

240
Q

OTHER MEDIA for ENTEROBACTERIACEAE

A

✓ MAC
✓ EMB
✓ Desoxycholate agars
✓ HEA
✓ XLD
✓ SSA
✓ Desoxycholate citrate agars
***Enrichment:
Selenite F
GN broth
Tetrathionate broth

241
Q

OTHER MEDIA for E. coli

A

✓MAC, EMB, XLD
✓SMAC

242
Q

OTHER MEDIA for Edwardsiella

A

MAC = colorless

243
Q

OTHER MEDIA for Shigella

A

✓ EMB, MAC, SSA = colorless
✓ XLD = red
✓ HEA = green to blue green

244
Q

OTHER MEDIA for Salmonella

A

✓ EMB, MAC, SSA = colorless
✓ XLD, BSA = black colonies w/ metallic
silver sheen
✓ Brilliant Green agar (BGA)
✓ HEA
Enrichment: Selenite, GNB

245
Q

OTHER MEDIA for Citrobacter

A

KCN medium

246
Q

OTHER MEDIA for K. pnemoniae

A

EMB, MAC, XLD = +string’s test

247
Q

OTHER MEDIA for Proteus

A

BAP = burnt gun odor , swarming

248
Q

OTHER MEDIA for Y. enterocolitica

A

CIN

249
Q

OTHER MEDIA for Pseudomonas aeruginosa

A

✓MHA
✓Pseudomonas P agar
✓Tech agar

250
Q

OTHER MEDIA for Eikenella

A

CAP = corrodes, pearly sheen (mercury droplet), bleach-like odor

251
Q

OTHER MEDIA for Flavobacterium

A

BAP = yellow

252
Q

OTHER MEDIA for Haemophilus

A

✓ Satellite phenomenon,
dewdrop like, bleachlike
✓ CAP
✓ Levinthal and Fildes
enriched media
✓ Horseblood bacitracin by
Klein and Blazevic

253
Q

OTHER MEDIA for Actinobacillus
actinomycetemcomitans

A

CAP = dots and dashes of Morse code

254
Q

OTHER MEDIA for Pasteurella

A

musty or mushroom-like odor (BAP)

255
Q

OTHER MEDIA for Francisella tularensis

A

✓Glucose cysteine blood agar
✓Peptone cysteine agar
✓Cystine heart agar
✓Chocolate agar
✓Rarely on Thayer Martin

256
Q

OTHER MEDIA for Brucella

A

✓ Castaneda biphasic medium
✓ W or Winsconsin medium
✓ Trypticase soy agar

257
Q

OTHER MEDIA for Bordetella

A

✓ Bordet-Gengou agar
✓ Jones-Kendrick charcoal agar
✓ Regan-Lowe
✓ BCYE
✓ cold casein hydrolysate
✓ Casamino acid broth
✓ Modified Stainer-Scholte agar with cyclodextrin and
cephalexin

258
Q

OTHER MEDIA for Kingella

A

BAP/CAP = “fried-egg” pitting appearance

259
Q

OTHER MEDIA for Vibrio cholerae

A

✓APW, TCBS,
✓Gohar, Dieudonne’s, Monsur and Aronson media

260
Q

OTHER MEDIA for Campylobacter jejuni

A

✓ Butzler’s medium
✓ Skirrow’s medium (also Helicobacter)
✓ Campy Thio medium
✓ Campy-BAP medium

261
Q

OTHER MEDIA for Leptospirae

A

✓Fletcher’s medium
✓Noguchi’s
✓Stewart’s
✓Ellinghausen, McCullough, Johnson and Harris (EMJH) medium

262
Q

OTHER MEDIA for Chlamydia trachomatis

A

✓McCoy cells
✓C. pneumoniae
✓HeLa 229

263
Q

OTHER MEDIA for Mycoplasma

A

✓SP-4 Mycoplasma medium
✓Edward-Hayflick agar
✓Shepard’s A-7B agar

264
Q

OTHER MEDIA for Rickettsia

A

✓Chick embryo

265
Q

OTHER MEDIA for Bartonella

A

✓ BHIA

266
Q

OTHER MEDIA for Legionella

A

✓ BCYE
✓ Feeley-Gorman medium
✓ MHA w/ hemin/isovitalex

267
Q

OTHER MEDIA for Chromobacterium

A

✓ smell of ammonium cyanide, violet in BAP

268
Q

OTHER MEDIA for Gardnerella vaginalis

A

✓ HBBT

269
Q

Ways to facilitate Anaerobic Cultivation

A

a. Special culture medium incorporated with Thioglycollate and Cystein (reducing agents)
b. Boiling of culture medium
c. Anaerobic chamber system with a vacuum pump and nitrogen gas to remove residual oxygen
d. Gas-pak jar conataining a palladium catalyst
e. Small volumes: plastic bags, pouches containing calcium carbonate and catalyst

270
Q

Specimen for Anaerobic Cultivations is aspirated with

A

sterile needle and syringe

271
Q

Anaerobic Transport Devices

A

➢Anaerobic Culturette
➢Bio-Bag
➢Anaerobic Pouch
➢GasPak Pouch

272
Q

A. Anaerobic Jars

A
  1. Brewer Jar
  2. Torbal Jar
  3. GasPak Jar
273
Q

Components of Anarobic Jars

A

➢Hydrogen and CO2 generator envelop→ activated with water
➢Palladium-coated alumina pellets→ catalyst
➢Methylene Blue or Resazurin Indicator Strip → upon exposure to atmospheric oxygen turns blue or pink

274
Q

→Oxygen is removed by means of electrically heated platinized catalyst with the electrical connection outside the jar

A

Brewer Jar

275
Q

→uses a rubber O ring rather than Plasticine and a catalyst active at room temperature thus requiring no electrical heating

A

Torbal Jar

276
Q

→most convenient and widely used anaerobic jar
→takes 30 to 45 minutes to obtain anaerobic
environment

A

GasPak Jar

277
Q

Principle of Anaerobic Jars

A

➢With water added to the CO2 and H2 generator envelop and oxygen catalysed with H2 to water via the pellets, anaerobiosis is achieve

278
Q

Indication of Anaerobiosis

A

✓Production of heat
✓Moisture inside jar
✓Decolorization of indicator strip: WHITE or COLORLESS

279
Q

(Anaerobic Jars)
Failure to achieve anaerobic condition will result to

A

“poisoned” catalyst or a crack in the O ring, jar or lid

280
Q

Jars utilizing the “Evacuation-Replacement” System

A

✓Air is removed by drawing a vacuum of 25 inches of mercury
✓Jar is filled with oxygen-free gas such as nitrogen between evacuations of air
✓Anaerobiosis is achieved more quickly with this method but less convenient for routine use

281
Q

→Enclosed sytem consist of
large clear plastic, airtight
bag or chamber filled with
oxygen-free gas mixture of
nitrogen, hydrogen and CO2
→Allow materials to enter
through an air lock

A

Glove Box or Anaerobic Chamber

282
Q

Anaerobiosis in Glove Box is maintained by

A

Palladium catalyst and Hydrogen gas

283
Q

Components of Glove Box

A

a. Nitrogen Gas
→ filler for the remaining percentage of the
anaerobic straucture
b. Palladium Pellets
→ remove residual oxygen by combining with
H2 to form water
c. Silica Gel (Dessicant)
→ absorbed water
d. Methylene Blue or Resazurin
→ oxygen reduction indicator

284
Q

Operator uses gloves or
sleeves that form airtight
seals around arms to
manipulate items

A

Glove Box

285
Q

→Pre-Reduced Anaerobically Sterilized (PRAS) agar is distributed under anerobic conditions as thin layer around the inner wall of test tubes
→tubes are rolled and cooled until the melted agar forms the thin layer

A

Role Tube Technique

286
Q

(Role Tube Technique)
→Syringe and needle are used through the rubber seal

A

Closed Method by Hungate

287
Q

(Role Tube Technique)
→ remove the rubber stopper and insert a cannula that has oxygen-free gas flowing from the tip

A

Open Method

288
Q

CULTURE MEDIA

A
  1. Freshly made BAP
  2. Enrichment media (supplemented BHIA, special Brucella Blood Agar, Laked Blood Agar)
  3. Selective Media
  4. Pre-Reduced Anaerobically Sterilized (PRAS)
    media
  5. Liquid Media
289
Q

Selective Media for anaerobic gram (-) rods

A

Kanamycin-Vancomycin Blood Agar

290
Q

Selective Media for Bacteroides melaninogenicus

A

Kanamycin-Vancomycin-Laked Blood Agar

291
Q

Selective Media for Fusobacterium
and Veilonella

A

Neomycin-Vancomycin Blood Agar

292
Q

Selective Media for Clostridia and anaerobic
gram(+) cocci

A

Neomycin Blood Agar

293
Q

Selective Media for Clostridia

A

Nagler Agar (with egg yolk, Neomycin)

294
Q

→ only medium capable of supporting
growth of 3 groups of microorganisms:
Aerobes, Microaerophiles, Anaerobes

A

Thioglycolate Medium

295
Q

→reducing substance, provides low redox potential

A

Na Thioglycollate

296
Q

→narrow pink layer means that the medium is reduced and can be used for anaerobic culture
→Pink layer extends to 1/3 of the medium: oxidized—boiled or autoclave to restore the narrow layer

A

Resazurin Indicator

297
Q

→stored at room temperature in the dark because at refrigerator temperature it absorbs more oxygen

A

Thioglycollate Medium

298
Q

Liquid Media that prevents entry of O2

A

Petrolatum

299
Q

Role Tube Technique is inoculated in what 2 ways

A

Close Method by Hungate
Open Method

BACTERIOLOGY - LAB (6 decks)

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