(F) Lesson 12: Haemophilus and Other Fastidious Bacteria (Part 2) Flashcards

1
Q
  • Composed of Haemophilus, Aggregatibacter, Cardiobacterium, Eikenella, Kingella
  • Most are part of the normal biota of the oral cavity and URT
  • All are related to bacterial endocarditis
  • Fastidious, facultative anaerobes, enhanced growth under presence of CO2
A

HACEK group

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2
Q

ACEK are ____ which means slower or poorer growing.

A

Dysgonic

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3
Q

T or F: The HACEK group includes those with immunocompromised state or those that are called as opportunistic pathogen.

A

T

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4
Q

The HACEK group are related to bacterial endocarditis due to their?

A

Predilection for attachment to heart valves

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5
Q

Familiarize yourself with the risk of infection for the HACEK group.

A
  • Tooth extraction
  • History of endocarditis
  • Gingival surgery
  • Heart valve surgery
  • Mitral valve prolapse
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6
Q
  • Comes from the Greek words aphros and philia which means foam-loving or desiring high concentration of CO2
  • Found in dental plaque and gingival scraping
  • Reclassification of H. aphrophilus and H. paraphrophilus due to molecular techniques
A

Aggregatibacter aphrophilus

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7
Q
  • Formerly part of actinobacillus
  • Divided into 6 serotypes: A to F
  • Etiologic pathogen for periodontitis
  • Has two major virulence factors: collagenase and leukotoxin
A

Aggregatibacter actinomycetemcomitans

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8
Q

Aggregatibacter actinomycetemcomitans

These serotypes (3) are the most common causative agents of infection.

A

A, B, C

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9
Q

Aggregatibacter actinomycetemcomitans

This virulence factor is for the destruction of collagen.

A

Collagenase

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10
Q

Aggregatibacter actinomycetemcomitans

This virulence factor is against WBCs and has something to do with anti-phagocytosis.

A

Leukotoxin

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11
Q

Colony of Aggregatibacter actinomycetemcomitans after 48 hours?

`

A

Star-shape with 4 to 6 points

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12
Q

T or F: The colony of Aggregatibacter actinomycetemcomitans may be observed with the naked eye.

A

F (use a microscope or stereomicroscope in LPO or scanner)

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13
Q

Drug of choice for Aggregatibacter actinomycetemcomitans?

A

Penicillin and an aminoglycoside (combined)

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14
Q

Alternative drugs for Aggregatibacter actinomycetemcomitans?

A
  • Third generation cephalosporins
  • Quinolones
  • Chloramphenicol
  • Tetracycline
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15
Q

Resistance of Aggregatibacter actinomycetemcomitans to ____ and ____ common.

A

Vancomycin and erythromycin

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16
Q

What are the two species of Cardiobacterium?

A
  • Cardiobacterium hominis
  • Cardiobacterium valvarum
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17
Q

Which Cardiobacterium spp. is more related to bacterial endocarditis?

A

Cardiobacterium hominis

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18
Q
  • Part of the normal biota
  • Has agar pitting
  • Infects the aortic valve more than other HACEK members
A

Cardiobacterium hominis

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19
Q

Drug of choice for Cardiobacterium hominis?

A

Penicillin with an aminoglycoside

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20
Q

Cardiobacterium hominis is sensitive to what drugs?

A
  • Beta-lactam antibiotics
  • Chloramphenicol
  • Tetracycline
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21
Q

Cardiobacterium hominis has a variable response to what drugs?

A
  • Aminoglycosides
  • Erythromycin
  • Clindamycin
  • Vancomycin
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22
Q
  • Part of the normal biota
  • Associated with human bites or fights (clenched fist wounds)
  • Opportunistic to immunocompromised individuals, producing periodontitis, meningitis, empyema, pneumonia, osteomyelitis, arthritis, and postoperative tissue infections
  • The least common isolate among the HACEK group for endocarditis
  • Called this because of corrosion seen in its agar pitting and yellow pigment
A

Eikenella corrodens

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23
Q

Eikenella corrodens is sensitive to what drugs?

A
  • Penicillin
  • Ampicillin
  • Cefoxitin
  • Chloramphenicol
  • Carbenicillin
  • Imipenem
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24
Q

Eikenella corrodens is resistant to what drugs?

A

Clindamycin and other narrow-spectrum cephalosporin

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25
Q
  • It resists in decolorization in G/S, appearing as G(+)
  • Major G(-) bacterium isolated from degenerative joint and bone infections (osteoarthritis) in children
A

Kingella kingae

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26
Q

What are the four members of the Kingella species?

A
  • Kingella kingae
  • Kingella denitrificans
  • Kingella oralis
  • Kingella potus
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27
Q

Which member of the Kingella species is related to endocarditis?

A

Kingella kingae

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28
Q

Kingella kingae’s ability to resist decolorizatuon is not due to what?

A

Peptidoglycan layer

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29
Q

Color of the colonies of Kingella kingae?

A

Yellow brown

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30
Q

T or F: Kingella kingae is resistant to most agents, including penicillin.

A

F (susceptible)

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31
Q

The five (5) identified species of Capnocytophaga spp. that are part of the oral microbiota

A
  • Capnocytophaga ochracea
  • Capnocytophaga gingivalis
  • Capnocytophaga sputigena
  • Capnocytophaga haemolyticus
  • Capnocytophaga granulosa
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32
Q

This species of Capnocytophaga spp. is the only one that tests negative for oxidase and catalse.

A

Capnocytophaga granulosa

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33
Q
  • Not as commonly involved in endocarditis as they are in septicemia
  • G/S: thin and often fusiform (pointed ends) resembling Fusobacterium spp.
  • Motile organisms with a gliding motility
  • On the agar: yellow-orange
A

Capnocytophaga spp.

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34
Q
  • Positive for: Sucrose, glucose, maltose, lactose, nitrite, and esculin
  • No growth on TSI without enrinchment
  • Non-hemolytic
A

Capnocytophaga spp.

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35
Q

This is the only beta-hemolytic organism from Capnocytophaga spp.

A

Capnocytophaga haemolytica

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36
Q

Capnocytophaga spp.

The two zoonotic species found in the cat/dog oral cavity from bite wounds.

A
  • Capnocytophaga canimorsus
  • Capnocytophaga cynodegmi
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37
Q

Drug of choice for Capnocytophaga canimorsus and Capnocytophaga cynodegmi?

A

Penicillin

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38
Q

The Capnocytophaga spp. are susceptible to what drugs?

A
  • Imipenem
  • Erythromycin
  • Clindamycin
  • Tetracycline
  • Chloramphenicol
  • Quinolones
  • Beta-lactams
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39
Q

The Capnocytophaga spp. are resistant to what drugs?

A

Aminoglycosides

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40
Q
  • Has 17 spp, but not all are related to man
  • (+) catalase, oxidase, glucose
  • (+) CHOC with grayish colonies
  • (-) MAC
  • (+) SBA (BAP) w/o satellitism, non-hemolytic, mucoid, narrow green to brown w/ halo around the colony
  • (+) bipolar staining in G/S with a safety pin appearance
A

Pasteurella spp.

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41
Q

What is the zoonosis of Pasteurella spp. and what are the involved organisms?

A

Systemic and cutaneous infections caused by Pasteurella canis, Pasteurella stomatis, and Pasteurella dagmatis

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42
Q

The most common isolate related to human infection involving the Pasteurella spp.

A

Pasteurella multocida

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43
Q

Familiarize yourself with the differential characteristics of Pasteurella spp.

A

Go mo na yan, it’s in the transes!

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44
Q

T or F: Most of the Pasteurella spp. are zoonotic, but due to their proximity to dogs and cats, humans can get infected.

A

T

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45
Q
  • The CDC categorizes them as category B agents since they can easily disseminate, cause moderate morbidity but low mortality
  • Under BSL-3 requiring many precautionary measures
  • Facultative intracellular pathogens
  • Colonies: smooth, raised, translucent
  • (+) Rose Bengal Test and 2-mercaptoethanol (2-ME) agglutination Test
A

Brucella spp.

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46
Q

What are the four members of the Brucella spp?

A
  • Brucella melitensis
  • Brucella abortus
  • Brucella suis
  • Brucella canis
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47
Q

Best media for Brucella spp.?

A
  • Castañeda broth
  • TSB (Trypticase Soy Broth)
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48
Q

Selective media for Brucella spp?

A

Wisconsin medium

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49
Q

This enhances the growth for Brucella spp.

A

Erythritol

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50
Q

What is the disease associated with the Brucella spp?

A

Brucellosis

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51
Q

Identify the clinical stage of Brucellosis.

Nonspecific symptoms (fever, malaise, headache, anorexia, myalgia, and back pain)

A

Acute

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52
Q

Identify the clinical stage of Brucellosis.

Undulating fevers (normal temp in the AM, high temp during PM), arthritis, and epididymoorchitis

A

Subchronic

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53
Q

Identify the clinical stage of Brucellosis.

Depression, arthritis, chronic fatigue syndrome

A

Chronic

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54
Q

Familiarize yourself with the differential characteristics of Brucella spp.

A

Nasa transes again hehe.

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55
Q

T or F: Natural hosts of Brucella spp. are usually humans.

A

F (animals, hence they are zoonotic)

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56
Q
  • Categorized by the CDC as Category A agents
  • Can pose a risk to national security because they can be spread through person-to-person contact or are easily disseminated and result in high mortality rates, leading to a potentially great public health impact and public panic
  • BSL-3
  • Fastidious, facultative intracellular
  • (-) oxidase, urease, satellite, X&V test
  • Weak (+) catalase and beta-lactamase activity
A

Francisella spp.

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57
Q

Francisella spp. requires the supplementation of what?

A

Cysteine, cystine, or thiosulfate

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58
Q

Familiarize yourself with the media used for Francisella spp.

A
  • CHOC
  • Modified Thayer-Martin
  • Buffered Charcoal Yeast Extract (BCYE) agar
  • Mueller-Hinton agar
  • Trypticase Soy Broths (TSB)
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59
Q

Familiarize yourself with the serologic tests used as diagnostic tools for infections relating to Francisella spp.

A
  • Direct fluorescent Ab (DFA)
  • Immunohistologic staining with monoclonal antibodies
  • PCR
  • Slide agglutination
  • Single serology test
60
Q

This remains the gold standard as isolation allows visualization of the causative agent, giving an ideal method of diagnosis.

A

Culture

61
Q

Francisella spp.

Most prevalent in human infection

A

Francisella tularensis

62
Q

What are the three subspecies of Francisella tularensis?

A
  • Tularensis (type A)
  • Holartica (type B)
  • Mediasiatica
63
Q
  • Also known as rabbit fever, deerfly fever, lemming fever or water rat trapper’s disease
  • MOT: ingestion, inhalation as pneumonia, arthropod ite in ulceroglandular form (by ticks, biting flies), or contact with infected tissue
A

Tularemia

64
Q

Has 52 spp with >70 serogroups wherein 26 are isolated from humans

A

Legionella spp.

65
Q

____ in Philadelphia in 1976 with 221 persons became ill with pneumonia-like symptoms wherein 34 of them died due to L. pneumophila.

A

American Legion Convention

66
Q
  • Agent of the outbreak
  • First named member of the family Legionellaceae
  • A primary human pathogen
  • 16 serogroups
  • 2 infections: Legionnaires’ Disease, Pontiac Fever depending on the manifestation of the disease
  • Categorized based on autofluorescence activity under UV light: yellow-green, blue-white, blue-white/yellow-green, no color
A

Legionella pneumophila

67
Q

Legionella spp.

  • Manifests as pneumonia
  • Top 4 causative agent of community-acquired bacterial pneumonia
  • Incubation: 2-10 days
  • (+) hemoptysis (blood is expectorated with saliva), rales (crackles when coughing), dyspnea (difficulty in breathing), and shaking chills
A

Legionnaires’ Disease

68
Q

What are the two types of pneumonia caused by Legionella pneumophila?

A

Typical and Atypical

69
Q

Type of Pneumonia from Legionella pneumophila

  • Progressive disease wherein signs and symptoms appear fast from the contact with the organism
  • Crackles when coughing with mucoid phlegm
A

Typical

70
Q

Type of Pneumonia from Legionella pneumophila

  • Slower and lesser symptoms presented
A

Atypical

71
Q

Legionella spp.

  • Non-pneumonic; more flu-like symptoms of fever, headache, and myalgia that lasts for 2-5 days
  • Incubation: 2 days
    Subsides without medical intervention (self-limiting)
A

Pontiac Fever

72
Q

T or F: Pontiac fever has more severe symptoms and manifestations and has a worse prognosis compared to Legionnaires’ Disease.

A

F (other way around)

73
Q

Specimens for Legionnaires’ Disease?

A
  • Sputum
  • Bronchoalveolar lavage
  • Bronchial washings

From respiratory tract

74
Q

T or F: Specimen that are stored in sterile and leak-proof containers are allowed in Legionella spp.

A

T

75
Q

If there is a delay for less than 2 hours for specimen of Legionella spp, what should be done?

A

Refrigerate at 2 to 8 deg C

76
Q

If there is a delay for several days for specimen of Legionella spp, what should be done?

A

Freeze at -70 deg C

77
Q

Specimen used for antigen detection for serological testing

A

Urine

78
Q

Result of microscopic examination for Legionella spp?

A

Weakly staining, gram-negative bacilli

79
Q

Specimen for isolation for Legionella spp.?

A

Sputum

80
Q

What should be done for sputum in isolation of Legionella spp?

A

Acid treatment with 1:10 with 0.2N KCl-HCl stood for 5 minutes

81
Q

Isolation of Legionella spp.

What is used to kill the normal microbiota?

A

0.2N KCl-HCl

82
Q

Media used for isolation of Legionella spp?

A

CHOC with L-cysteine, BCYE

83
Q

Best medium for isolating Legionella spp.

A

BCYE agar with L-cysteine

84
Q
  • Appear grayish-white to bluish-green, convex, and glistening (shining)
  • Central portion of young colonies appears light gray and granular w/ ground glass appearance
  • The periphery of the colony appears pink or light blue or bottle green with a furrowed appearance
A

Legionella spp.

85
Q

What are the three conventional methods for Legionella spp?

A
  • Gram-stain
  • L-cysteine
  • DFA Test
86
Q

Conventional Methods for Legionella spp.

  • For any suspicious colony growth on BCYE agar
  • Legionella spp. are thin G(-) bacilli
  • Spx from LRT
A

Gram-stain

87
Q

Conventional Methods for Legionella spp.

  • From suspected colonies
  • (+) SBA or BCYE with L-cysteine
  • (-) SBA or BCYE w/o L-Cysteine
A

L-cysteine

88
Q

Conventional Methods of Legionella spp.

  • A serological test
  • Prepare smears from colonies that require L-cysteine and lay over with polyvalent (Pseudomonas, Bacteroides, Corynebacterium) and monovalent conjugates to determine specific species and serogroup
A

DFA Test

89
Q

What are the rapid methods for Legionella spp?

A
  • Urine Antigen Test
  • DNA Detection
  • DFA Test
90
Q

Rapid Methods for Legionella spp.

  • Spx: urine
  • Methods: radioimmunoassay, microplate enzyme immunoassay, and rapid immunochromatographic assay
  • (+) infection with Legionella
A

Urine Antigen Test

91
Q

Rapid Methods for Legionella spp.

  • PCR for nucleic acid amplification
  • Well-refined MolBio procedures
A

DNA Detection

92
Q

Rapid Methods of Legionella spp.

  • Detection of more common species of Legionella
  • Conjugate + fluorescein isothiocyanate (fluorochrome)
  • Uses fluorescence microscope
  • (+) bright yellow to green, short or coccobacillary bacilli with intense peripheral staining
A

DFA Test

93
Q

Drug of choice for Legionella spp. in early dignosis?

A

Azithromycin or fluoroquinolone

94
Q

Alternative drug for Legionella spp. in early diagnosis?

A

Doxycycline

95
Q
  • There are 8 known spp and 2 are related to humans which are primary pathogens while the others are zoonotic
  • G(-) bacilli or coccobacilli
  • Fastidious, obligate aerobe
  • (+) catalase, oxidize, amino acid
  • (-) carbohydrate fermentation
A

Bordetella spp.

96
Q

Growth of Bordetella spp. is inhibited by?

A
  • FA
  • Metal ions
  • Sulfides and peroxides
97
Q

Protective substances needed Bordetella spp. to counteract the inhibitors?

A
  • Charcoal
  • Blood
  • Starch
98
Q

Species of Bordetella spp.

Primary human pathogen

Two (2)

A
  • B. pertussis
  • B. parapertussis
99
Q

Species of Bordetella spp.

Respiratory tract pathogens of wild and domestic birds and mammals

A
  • B. bronchiseptica
  • B. avium
100
Q

Species of Bordetella spp.

Opportunistic human pathogen

A

B. bronchiseptica

101
Q

Species of Bordetella spp.

Bacteremia among immunocompromised

A

B. holmesii

102
Q

Species of Bordetella spp.

Ear infection in immunocompromised

A

B. trematum

103
Q

Species of Bordetella spp.

Other species?

A
  • B. hinzii
  • B. petrii
104
Q

Virulence Factors of Bordetella spp.

  • Facilitates attachment to ciliated epithelial cells
A

Filamentous Hemagglutinin (FHA)

105
Q

Virulence Factors of Bordetella spp.

  • Protein exotoxin that produces a wide variety of responses in vivo
  • B. parapertussis and bronchiseptica contain structural genes for PT but do not express the complete operon
A

Pertussis Toxin

106
Q

Virulence Factors of Bordetella spp.

Modifies host proteins which interfere with signal transduction between cells and phagocytes

A

Adenosine diphosphate-ribosyl transferase

107
Q

Virulence Factors of Bordetella spp.

  • Inhibits host epithelial and immune effector cells by inducing supraphysiologic concentrations of cyclic adenosine monophosphate
  • Immune system is silenced
A

Adenylate Cyclase Toxin

108
Q

VIrulence Factors of Bordetella spp.

  • Causing ciliostasis, inhibiting DNA synthesis, and promoting cell death
A

Tracheal Cytotoxin

109
Q

Clinical Manifestations of Bordetella spp.

  • 1-3 weeks after incubation
  • Symptoms: insidious, non-specific: sneezing, mild cough, runny nose, conjunctivitis
  • Highly communicate because of the large number of organisms in the respiratory tract
  • Cultures are not often performed
A

Catarrhal Phase

110
Q

Clinical Manifestations of Bordetella spp.

  • Severe, repetitive coughing with a “whoop” at the end of the coughing spell
  • Young children: pneumonia, or both and require aid in maintaining a patent airway
  • B. parapertussis presents a similar disease but with milder symptoms
A

Paroxysmal Phase

111
Q

Clinical Manifestations of Bordetella spp.

There is a wheezing sound created during an attempt to grasp air in between coughs, which is a characteristic of Bordetella pertussis.

A

Whooping cough

112
Q

Clinical Manifestations of Bordetella spp.

Target of pertussis?

A

Young children

113
Q

Clinical Manifestations of Bordetella spp.

  • After surviving the catarrhal and paroxysmal stages
  • 4 weeks after onset with decreased frequency and severity of coughing spells
  • Complete recovery can take weeks or months
A

Convalescent Phase

114
Q
  • Includes diphtheria, tetanus toxoid, acellular pertussis
  • Requires booster
A

DTaP vaccine

115
Q

Bordetella spp.

  • Kennel cough in dogs
  • Can cause parvo
A

B. brochiseptica

116
Q

Bordetella spp.

Infrequently associated with pertussis syndrome and other RT infection

A
  • B. bronchiseptica
  • B. holmesii
117
Q

Bordetella spp.

Commonly associated with pertussis

A
  • B. pertussis
  • B. parapertussis
118
Q

Specimen of choice for Bordetella spp?

A

Nasopharyngeal swab or aspirate

119
Q

Specimen Collection and Handling for Bordetella spp.

These are more preferred as they easily available and easy to prepare.

A

Swabs

120
Q

Specimen Collection and Handling for Bordetella spp.

These have higher sensitivity when attempting to recover respiratory pathogens.

A

Flocked swabs

121
Q

T or F: Throat cultures are less sensitive, hence discouraged.

A

T

122
Q

Transport System for Bordetella spp.

Less than two hour delay?

A

1% casein hydrolysate (casamino acids) broth at RT

123
Q

Transport System for Bordetella spp.

24-hour delay?

A

Amies transport medium with charcoal at RT

124
Q

Transport System for Bordetella spp.

Overnight or several day delay?

A

Regan-Lowe Transport Medium

Containing half-strength charcoal agar containing 10% horse blood and 40 mg/L cephalexin at 350C

125
Q
  • Primary rapid diagnostic strategy
  • Circumvent many of the problems associated with specimen transport and bacterial cultivation
  • Uses at least two DNA targets (e.g., IS481 and pxtS1)
  • May need to seek confirmation to avoid the mischaracterization of respiratory illness
A

Nucleic Acid Detection

126
Q

Microscopic examination for Bordetella spp?

A

DFA test + culture

127
Q

Microscopic Examination of Bordetella spp.

  • Uses polyclonal fluorescent labeled conjugates for both B. pertussis and B. parapertussis
  • DFA lacks sensitivity
  • False positive result
A

DFA test + culture

128
Q

Bordetella spp. are incubated at what conditions?

A

35 deg C in ambient air for 7 days minimum

129
Q

This is colony checking for Bordetella spp.

A

Stereomicroscope

130
Q

Familiarize yourself with the media used for Bordetella spp.

A
  • Bordet-Gengou potato infusion agar with glycerol and horse or sheep blood
  • Charcoal agar supplemented with 10% horse blood and 40 mg/L cephalexin
  • Regan and Lowe
  • Jones Kendrich: charcoal and yeast extract
  • Charcoal cephalexin blood agar (CCBA)
  • Stainer and Scholte broth
  • Casamino broth
131
Q

Young or old colony of Bordetella spp.

  • Smooth, glistening, and silver, resembling mercury droplets
A

Young colonies

132
Q

Young or old colony of Bordetella spp.

  • Turn whitish-grey
A

Older colonies

133
Q

ID Methods of Bordetella spp.

  • Tiny G(-) cocco bacilli (elongated if from cephalexin-containing media)
  • May need prolonged safranin exposure of up to 2 mins to see typical morphology
A

Gram-Stain

134
Q

ID Methods of Bordetella spp.

  • Exhibit bipolar granules of Bordetella
A

Toluidine Blue Bipolar Granules

135
Q

ID Methods of Bordetella spp.

  • For agglutination test or fluorescein-labeled antisera
  • Both (+) = no more confirmatory test = B. pertussis
  • Equivocal results = subculture to charcoal horse blood agar (CHBA), SBA, CHOC for biochemical testing
A

Suspicious Colonies

136
Q

ID Methods of Bordetella spp.

  • One of the most promising equipment
  • Mass spectrometry and 166 rRNA gene sequencing
  • For accurate ID
A

Matrix-Assisted Laser Desorption/Ionization Time Of Flight (MALDI-TOF)

137
Q
  • Assays cannot distinguish immune response between infection or vaccination
  • Whether it is a current or past infection
  • Cannot ID B. parapertussis vs B. pertussis infection
  • Foro detecting Ab and Ag
A

Serologic Testing

138
Q

Serologic Testing of Bordetella spp.

  • Enzyme immunoassay (EIA) or Bead-based assay
  • For diagnosis exposure to B. pertussis
A

Antipertussis Toxin AB

139
Q

Serologic Testing of Bordetella spp.

  • More ideal
  • 100 IU/mL: recent infection
  • >40 IU/mL: looking for past infection or exposure
  • There is difficulty in diagnosis
A

IgG Antibody Titer

140
Q

Serologic Testing of Bordetella spp.

  • Not recommended, rarely done
A

IgM Antibody Titer

141
Q
  • Sensitive to macrolides, ketolides, penicillin, tetracycline
  • Resistant to oral cephalosporins
A

B. pertussis and B. parapertussis

142
Q

Antimicrobial Susceptibility of Bordetella spp.

  • DOC for treatment and prophylaxis
  • Efficient if treatment begins during the catarrhal phase
  • Some may exhibit resistance
  • Determine if it can still be used via AST
A

Erythromycin

143
Q

Antimicrobial Susceptibiity of Bordetella spp.

  • Longer half-life
  • High patient compliance due to fewer and milder side effects with fewer daily doses
A

Azithromycin

144
Q

Antimicrobial Susceptibility of Bordetella spp.

  • Alternative for treatment and prophylaxis in place of erythromycin
A

Trimethoprim-sulfamethoxazole

145
Q
  • Unpredictable susceptibility
  • Usually susceptible to aminoglycosides
A

B. bronchiseptica