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Clinical Bacteriology > (M) Lesson 8.1: Mycobacteria (Part 1) > Flashcards

(M) Lesson 8.1: Mycobacteria (Part 1) Flashcards

1
Q
  • Slender, slightly curved or straight rods
  • Non-motile, strict aerobes
A

Mycobacterium

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2
Q

All Myconacterium are non-sporeforming except?

A

Mycobacterium marinum

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3
Q

All Mycobacterium are slow growers except?

A
  • Mycobacterium fortuitum
  • Mycobacterium chelonae
  • Mycobacterium phlei
  • Mycobacterium stegmatis
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4
Q

60% of the cell wall contains what?

Clue: Two answers

A
  • High lipid content
  • Lipid containing structures (Mycolic Acid and Cord Factor Wax D)
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5
Q

Mycobacterium is also referred to as?

A

Acid Fast Bacilli

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6
Q
  • Characteristic or main feature of Mycobacteria
  • Pertains to the ability of the bacterial cell wall to withstand strong acid decolorizer due to the presence of long chain waxy mycolic acid
A

Acid Fastness

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7
Q

Mycobacterium contains ____ instead of N-acetylmuramic acid

A

N-glucolyl muramic acid

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8
Q

T or F: There is low lipid content in Mycobacterium - which is why they are able to create a hydrophobic permeability barrier.

A

F (very high lipid content)

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9
Q

The reason as to why Mycobacteria grow more slowly (the organisms tend to clump together and the nutrients are not easily allowed into the cells)

A

Hydrophobicity

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10
Q

Classify if MTC or NTM

  • M. tuberculosis
  • M. bovis
  • Mbovis BCG
  • M. africanum
  • M. caprae
  • M. microti
  • M. canetti
  • M. mungi
  • M. oygis
  • M. pinnipedii
A

Mycobacterium tuberculosis complex (MTC)

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11
Q

Classify if MTC or NTM

Slow-growing pathogenic
* M. avian complex

Rapid-growing opportunistic pathogenic
* M.aviancomplex
* M. fortuitum group
* M.chelonae
* M.abssessus subs. Abscessus

Non-cultivable NTM
* M. leprae

A

Non-tuberculosis mycobacteria (NTM)

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12
Q
  • Complex: Two or more species that are difficult to distinguish and have little medical importance
  • Can cause tuberculosis
  • Non-pigmented colonies
  • Slow grower
A

Mycobacterium tuberculosis complex

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13
Q

How many days does it require for Mycobacterium tuberculosis complex to grow?

A

More than 7 days (2 to 60 days)

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14
Q

Identify the organism based on habitat and mode of transmission.

  • Habitat: Patient with cavitary (lung) disease (primary reservoir)
  • Mode of Transmission: P2P, ABT (single bacilli may cause infection, only 15 to 20% develop diseases years later)
A

M. tuberculosis

ABT - Airborn Transmission
P2P - Person to Person

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15
Q

Identify the organism based on habitat and mode of transmission.

  • Habitat: Humans and animals
  • Mode of Transmission: Ingestion of contaminated milk, ABT
A

M. bovis

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16
Q

Identify the organism based on habitat and mode of transmission.

  • Habitat: Commercial vaccines
  • Mode of Transmission: Via vaccination (rare)
A

M. bovis BCG

BCG - Bacillus Calmette-Guerin

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17
Q

Identify the organism based on habitat and mode of transmission.

  • Habitat: Humans
  • Mode of Transmission: ABT
A

M. africanum

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18
Q

Identify the organism based on habitat and mode of transmission.

  • Habitat: Animals, Humans (rarely)
  • Mode of Transmission: ABT

Clue: Three answers

A
  • M. caprae
  • M. microti
  • M. pinnipedii
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19
Q

Hi idol, study the epidemiology of TB and TB-HIV !!

A

Go na please mwa

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20
Q

Ghon’s complex
* Focus of inflammation
* Pulmonary lesion caused by tuberculosis

A

Primary (Initial) Pulmonary Tuberculosis

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21
Q
  • Also called reactivation
A

Cavity (Secondary) Pulmonary Tuberculosis

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22
Q

Cavity (Secondary) Pulmonary Tuberculosis

Hard tubercle in lung-cellular aggregate

A

Granuloma

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23
Q

Cavity (Secondary) Pulmonary Tuberculosis

Cheese-like dead tissue to macrophage enzymes

A

Caseous necrosis

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24
Q
  • In patients infected with primary active TB, the disease may be spread through lymphatic system
  • Progress to form cavities in lungs and other organs
  • Rarely do granulomas become calcified and remain asymptomatic for years
A

Miliary (Disseminated) Pulmonary Tuberculosis

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25
Q

Common name of Mycobacterium tuberculosis

A

Koch bacillus

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26
Q

Who discovered the Mycobacterium tuberculosis?

A

Robert Koch (1882)

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27
Q
  • Obligate aerobe
  • Require CO2 for growth (5 to 10%)
  • Virulence factor
A

Mycobacterium tuberculosis

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28
Q

Characteristics of Mycobacterium tuberculosis

Sticky response of the sputum

A

Cord Factor and Sulfatides

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29
Q

Disease

  • The TB bacilli are inside the body but they cannot spread the disease to others
  • However, if the patient do not get the treatment needed it may develop to Active TB
A

Latent TB infection

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30
Q

Disease

  • It can spread the disease from one person to another
  • Can cause death if not treated immediately
  • The bacilli grows in the body and makes the person feel sick and have symptoms
A

Active TB Disease

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31
Q

Disease

What are the possible symptoms for TB?

A
  • Productive cough
  • Low grade fever
  • Myalgia
  • Chills
  • Sweating
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32
Q

Disease

  • Happens when the immune of an individual becomes weak because the body cannot control the bacteria from growing because the immune system is suppressed
  • Can also mimic other diseases
A

Reactivation Tuberculosis

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33
Q

Disease

What are the diseases that TB can mimic?

A
  • Influenza
  • Acute Bronchitis
  • Pneumonia
  • Neoplasm
  • Fungal Infections
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34
Q

The primary means of identifying individuals affected by mycobacterium

A

Tuberculin Skin Test

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35
Q
  • Purified Protein Derivative (PPD)
  • 2-4 Days: Delayed Hypersensitivity Reaction (Erythema)
A

Tuberculin Skin Test

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36
Q
  • A culture extract of mycobacterium tuberculosis is injected intramuscularly or intracutaneously
  • After 48-72 hours, the individual will show delayed hypersensitivity reaction to PPD and it is characterized by erythema or redness and also in duration or firmness as a result of the influx of the immune cells
  • Not 100% specific or sensitive
A

Tuberculin Skin Test

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37
Q
  • Blood incubated with Mtb Ag
  • Measure released cytokines using enzyme-linked immunospot
A

T-Spot TB Test

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38
Q
  • Measure T-cells activated by MTb Ag
  • Should be interpreted in correlation with the patient’s signs and symptoms
A

T-Spot TB Test

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39
Q
  • Enzyme-Linked Immunoassay (ELISA)
  • Detects interferon-γ released from lymphocytes
A

Quantiferon-TB Gold

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40
Q
  • Detects the interference gamma that is release from the lymphocytes
  • Another important feature is that the result of the assay are not affected by the BCG vaccination
A

Quantiferon-TB Gold

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41
Q

What the different types of specimen for Mycobacterium tuberculosis?

A
  • Sputum (Expectorated and Induced)
  • Blood
  • Urine
  • CSF
  • Pleural, Pericardial, and Peritoneal Fluid
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42
Q

The most common specimen for Mycobacterium tuberculosis?

A

Sputum

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43
Q

Sputum specimen must be collected over the course of?

A

Three (3) consecutive days

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44
Q

Types of Sputum

  • Deep cough
A

Expectorated Sputum

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45
Q

Types of Sputum

  • Through inhalation of NaCl using nebulizer
  • Obtained through bronchoscopy
A

Induced Sputum

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46
Q

Sputum secretions are obtained by?

A

Bronchoscopy

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47
Q

Gram stain quality specimen is classified with?

A

Bartletts Classification

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48
Q

Saliva or sputum?

Greater than 10 EC with 25 Polymorphonuclear (PMN)

A

Saliva (rejected)

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49
Q

Saliva or sputum?

Less than 10 EC with less than 25 Polymorphonuclear (PMN)

A

Sputum (accepted)

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50
Q

CSF (Specimen for M. tuberculosis)

  • Pellicle of web-like clot (positive in TB) indicates what?
A

Tubercular meningitis

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51
Q

T or F: There is increased ADA in Mycobacterium tuberculosis.

A

T

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52
Q

Produced by lymphocytes which is a useful marker for the diagnosis of tuberculous pleurisy

A

Adenosine Deaminase

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53
Q
  • Pleural effusion secondary to infection of mycobacteria
A

Tuberculous pleurisy

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54
Q
  • Presumptive for MTb
  • Microscopic examination of specially stained smears to detect acid fast organism
A

AFB Smear Microscopy (DSSM)

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55
Q

What are the two different types of smears?

A
  • Direct Smear
  • Concentrated Smear
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56
Q

Types of Smears

  • Prepare directly from a patient specimen prior to processing
A

Direct Smear

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57
Q

Types of Smear

  • Prepared from a processed specimen after centrifugation to concentrate the material
A

Concentrated Smear

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58
Q

Identify the method.

  • Carbol Fuchsin with heat
  • Acid Alcohol (3% HCl in 95% ethanol
  • Methylene Blue
A

Ziehl-Neelsen’s Method (Hot Method)

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59
Q

What is the mordant for Ziehl’s Neelsen’s Method?

A

Heat

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60
Q

Ziehl-Neelsen’s Method

Blue bacilli against blue background is reported as?

A

Non-Acid Fast

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61
Q

Ziehl-Neelsen’s Method

Red bacilli against blue background is reported as?

A

Acid Fast Bacilli

62
Q

Identify the method.

  • Carbol Fuchsin (no heat)
  • Acid-alcohol 0.5% HCl in 95% alcohol
  • Malachite Green
A

Kinyoun’s Method (Cold Method)

63
Q

What is the mordant chemical of Kinyoun’s Method?

A

Tergitol

64
Q

Kinyoun’s Method

Red bacilli against green background

A

Acid Fast Bacilli

65
Q

Kinyoun’s Method

Green bacilli against green background

A

Non-Acid Fast

66
Q

Identify the method.

  • Carbol Fuchsin (with heat)
  • Pappenheim’s differentiating stain
  • Rosalic acid with Methylene Blue in glycerine and absolute alcohol
A

Pappenheim’s Method

67
Q

Pappenheim’s Method

Red or pink bacilli is reported as?

A

M. tuberculosis

68
Q

Pappenheim’s Method

Colorless bacilli is reported as?

A

M. stegmatis

69
Q

Identify the method.

  • Alcoholic Carbol Fuchsin
  • 1% nitric acid in 95% alcohol
A

Baumgarten’s Method

70
Q

Baumgarten’s Method

Blue bacilli is reported as?

A

M. tuberculosis

71
Q

Baumgarten’s Method

Red bacillis is reported as?

A

M. leprae

72
Q

Identify the method.

  • Alcoholic Carbol Fuchsin
  • Acid Alcohol (3% HCl in 95% ethanol)
  • Gabbet’s Methylene Blue Solution
A

Gabbet Method

73
Q

Gabbet Method

Red bacilli against blue background is reported as?

A

Acid Fast Bacilli

74
Q

Gabbet Method

Blue bacilli against blue background is reported as?

A

Non-Acid Fast

75
Q

T or F: A positive smear will provide a conclusive and vital clue for the clinician in the diagnosis of Tuberculosis.

A

F (presumptive not conclusive)

76
Q

T or F: The acid fast stain can help in monitoring the course of treatment.

A

T

77
Q
  • The carbol fuchsin binds to the mycolic acid in the cell wall of the mycobacteria
  • The fuchsin and the phenol are driven into the cell wall of the microorganism
A

Ziehl-Neelsen Method

78
Q
  • The tergitol forces the fuchsin into the cell wall
A

Kinyoun Technique

79
Q

This acts as a counterstain for either hot or cold procedure.

A

Methylene Blue

80
Q
  • Its differentiating stain contains Rosalic acid, methylene blue, glycerin, absolute alcohol
  • Differentiates M. stegmatis from M. tuberculosis
A

Pappenheim’s Method

81
Q
  • Differentiates M. leprae from M. tuberculosis
A

Baumgarten’s Method

82
Q
  • Contains methylene blue, sulfuric acid, absolute alcohol, and distilled water
A

Gabbet’s Methylene Blue Solution

83
Q

Gabbet’s Methylene Blue Solution

Acts as the primary stain

A

Alcoholic carbol fuchsin

84
Q

Gabbet’s Methylene Blue Solution

Acts as the decolorizer

A

Acid alcohol (3% HCl, 95% ethanol)

85
Q

Gabbet’s Methylene Blue Solution

T or F: The interpretation is same as ZN method.

A

T

86
Q

Gold standard method for TB diagnosis

A

TB Culture

87
Q

Used for species identification, drug susceptibility testing, and genotyping

A

Culture

88
Q

Two major categories of culture media

A
  • Solid (Egg and Agar based)
  • Liquid (Broth media)
89
Q

Solid Media Comparison

  • Selective: Malachite green
  • Rate of Growth: Slower
  • Area of Inoculum: Usually less but if present involves the entire surface
  • Visual Examination: Difficult due to media opacity
  • CO2: Not required
  • DST: Not performed in LJ
  • Shelf Life: Longer (6 to 12 months)
A

Egg-based Media

90
Q

Solid Media Comparison

  • Selective: Added antibacterial and antifungal
  • Rate of Growth: Faster
  • Area of Inoculum: More frequent, but can still isolate colonies
  • Visual Examination: Media is clear, allows easier colony quantification and morphology confirmation
  • CO2: Required
  • DST: Preferred due to larger surface area and faster rate of growth
  • Shelf Life: Shorter (1 to 2 months)
A

Agar-based Media

91
Q

Solid Agar Based

  • For Mycobacteria isolation
A
  • Middlebrook 7H10
  • Middlebrook 7H11
92
Q

Solid Agar Based

  • For Mycobacteria isolation from heavily contaminated cultures
A

Mitchison’s selective

93
Q

Solid Egg Based

  • For isolation and cultivation of Mycobacteria
A
  • Wallenstein
  • Lowenstein-Jensen
  • L-J pyruvic acid
94
Q

Solid Egg Based

  • Contains higher concentration of malachite green
A

Petragani Medium

95
Q

Liquid Based

  • Automated System
A
  • Bactec MGIT
  • VERSATREK
  • BacT/ALERT 3D
96
Q

TB colonies resemble what?

A

Cauliflower-like colonies

97
Q

Why do we need the methods of decontamination of bacteria?

A
  • Most specimen are contaminated with normal flora and can overgrow the slow-growing Mycobacteria
  • To isolate the Mycobacterium
98
Q

T or F: Specimen like CSF must be sterile.

A

F (sterile already since they are collected aseptically)

99
Q

Involves splitting the disulfide bonds in mucin that may trap bacteria

A

Digestion of sputum samples

100
Q

Identify the reagent.

  • Digestant and decontaminating agent
A
  • 2-4% NaOH
  • Benzalkonium chloride (Zephiran)
101
Q

Identify the reagent.

  • Liquefying agent
A

N-acetyl-L-cysteine (NALC)

102
Q

Identify the reagent.

  • Decontaminates specimen contained with P. aeruginosa
A

5 to 6% Oxalic Acid

103
Q

Biochemical Tests

  • Reagent strips impregnated with cyanogen bromide are inoculated with the organism
  • Principle: Niacin + Niacin ribonucleotide + aniline dye + cyanogen bromide
  • For M. tuberculosis identification
A

Niacin Test

104
Q

Niacin Test

Positive result?

A

Yellow (accumulation of free niacin - lack of the enzyme)

105
Q

Niacin Test

These species produce a positive result for Niacin Test.

A
  • M. tuberculosis
  • M. simiae
  • M. marinum
  • M. chelonae
106
Q

Niacin Test

Negative results?

A

No color change

107
Q

Niacin Test

This specie is the only one that produces a negative result.

A

M. bovis

108
Q

Biochemical Tests

T or F: Most mycobacterium species are catalase positive.

A

T

109
Q

Catalase Test at 68C or Heat-Stable Catalase

Positive results?

A

45 mm gas bubble height

110
Q

Catalase Test at 68C or Heat-Stable Catalase

Negative results?

A

No bubbles

111
Q

Catalase Test at 68C or Heat-Stable Catalase

The only specie that produces a positive result.

A

M. kansasii

112
Q

Catalase Test at 68C or Heat-Stable Catalase

Species that produce negative results.

A

All members of the MTB complex react negatively.

113
Q

Biochemical Test

  • Mycobacteria possess the enzyme nitroreductase that converts nitrates to nitrites
  • Principle: HCl + sulfanilamide + N-naphthylethylene diamine
A

Nitrate Reduction Test

114
Q

Nitrate Reduction Test

Positive results?

A

Pink or red

115
Q

Nitrate Reduction Test

Species that produce a positive result.

A
  • M. kansasii
  • M. szulgai
  • M. fortuitum
  • M. tuberculosis
116
Q

Nitrate Reduction Test

Negative results?

A

No color change

117
Q

Nitrate Reduction Test

Species that produce a negative result.

A
  • M. avium-intracellulare complex
  • M. bovis
  • M. marinum
  • M. simae
  • M. gastri
118
Q

Biochemical Tests

  • Principle: Tween 80 converted to oleic acid by Tween 80 lipase
  • Useful for identification of M. kansasii as quick as 6 hours
  • Differentiates M. gordonae and M. scrofulaceum
A

Tween 80 Hydrolysis

119
Q

Tween 80 Hydrolysis

Positive results?

A

Red color

120
Q

Tween 80 Hydrolysis

Species that produce a positive result.

A
  • M. kansasii
  • M. gordonae
121
Q

Tween 80 Hydrolysis

Negative results?

A

No red/amber

122
Q

Tween 80 Hydrolysis

Species that produce a negative result.

A
  • M. avium
  • M. scrofulaceum
123
Q

Biochemical Tests

  • Principle: Certain mycobacteria can reduce tellurite salts to metallic tellurium forming a black precipitate
A

Tellurite Reduction Test

124
Q

Telleurite Reduction Test

Positive results?

A

Smooth fine black precipitate

125
Q

Tellurite Reduction Test

Specie that produces a positive result.

A

M. avium

126
Q

Tellurite Reduction Test

Negative results?

A

Gray clumps

127
Q

Tellurite Reduction Test

Specie that produces a negative result.

A

M. kansasii

128
Q

Biochemical Tests

  • For rapid growers
  • Principle: Aryl sulfatase split sulfate group from tripotassium phenolphthalein disulfide/sulfate, producing free phenolphthalein
  • For identification of M. fortuitum-chelonae complex that gives (+) result after 3 days
A

Aryl Sulfatase Test

129
Q

Aryl Sufatase test

Positive results?

A

Pink or red color

130
Q

Aryl Sulfatase Test

Specie that produces a positive result.

A

M. fortuitum-chelonae complex

131
Q

Biochemical Tests

  • A susceptibility test
  • Casein hydrolysis serves as a growth stimulant for drug-resistant strains of M. tuberculosis
A

Thiphene-2-Carboxylic Acid Hydralize (TCH) Test

132
Q

TCH Test

Components of the TCH Agar

A
  • Inorganic salt essential (for mycobacterial growth)
  • Glycerol(source of carbon and energy)
133
Q

TCH Test

This serves as a growth stimulant for drug-resistant strains of M. tuberculosis.

A

Casein hydrolysis

134
Q

TCH Test

Susceptible to TCH Test

A

M. bovis

135
Q

TCH Test

Resistant to TCH Test

A

M. tuberculosis

136
Q

Biochemical Tests

  • Only rapid growers can convert 20% ferric ammonium citrate to an iron oxide
A

Iron Uptake

137
Q

Iron Uptake

Positive results?

A

Rusty brown colonies

138
Q

Iron Uptake

Species that produce a positive result.

A

M. phlei, M. smegmatis (rapid growers)

139
Q

Iron Uptake

Negative results?

A

No color formation

140
Q

Iron Uptake

Specie that produces a negative result.

A

M. chelonae (also a rapid grower)

141
Q

Biochemical Tests

Converts pyrazinamide to pyrazinoic acid and ammonia in 4 days

A

Pyrazinamidase

142
Q

Pyrazinamidase

Positive result?

A

Red pigment

143
Q

Pyrazinamidase

Species that produces positive results.

A
  • M. tuberculosis
  • M. marinum
144
Q

Pyrazinamidase

Negative results?

A

No color formation

145
Q

Pyrazinamidase

Species that produces a negative result.

A
  • M. bovis
  • M. kansasii
146
Q

Biochemical Tests

  • The detection of urease activity
A

Urease

147
Q

Urease

Specie that produce positive results.

A

M. scrofulaceum

148
Q

Urease

Specie that produces negative results.

A

M. gordonae

149
Q

Biochemical Tests

T or F: Most bacteria cannot grow in high salt concentration, 5% NaCl in egg-based media, inhibiting the growth of most media

A

T

150
Q

Biochemical Tests

  • For identification of M. triviale, the only slow grower that gives (+)
A

Sodium Chloride Tolerance

151
Q

Sodium Chloride Tolerance

Species that gives positive results (growth)?

A
  • M. flavescens
  • M. trivale
  • Mycobacterium rapid growers

Clinical Bacteriology (33 decks)

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