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Medical Microbiology II - 4336 > Test 2: Anaerobes > Flashcards

Test 2: Anaerobes Flashcards

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1
Q

List four obligate anaerobes and what percent oxygen

A

Prevotella sp.
Fusobacterium sp.
Bacteroides spp.
Clostridium perfringens

-less than 0.5% oxygen

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2
Q

What is the normal habitats of anaerobes?

A

Mucosal surfaces, bowel, skin

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3
Q

Where are anaerobes part of the normal microbiota?

A

Feces, skin, vagina, mouth, upper respiratory.

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4
Q

Anaerobes outnumber aerobes ______ to 1 in feces,
____ to 1 in in skin, mouth and vagina.

A

1000

10

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5
Q

Indigenous microflora of the colon:

Majority of bacteria are ___________ with a small amount of facultative organisms.

A

obligate anaerobes

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6
Q

Clues to anaerobic infections?

A

*A foul odor emanating from the specimen
*A lesion close to a mucosal surface
*The presence of an underlying disease:
tissue necrosis, impaired blood supply
*Previous antibiotic therapy
*Infection following a bite wound
*Sulfur granules exudating from the lesion

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7
Q

Vincent’s angina is now called…

A

Anaerobic tonsillitis

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8
Q

Types of infections of the head, throat, mouth, and repiratory?

A

-Anaerobic tonsillitis
-Ludwig’s angina
-Otitis media and sinusitis
-Aspiration pneumonia
-Cervico-facial actinomyces

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9
Q

What bacteria causes acne?

A

Propionibacterium acnes

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10
Q

Ture or false,

anaerobic infections are common in bite wounds.

A

True

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11
Q

Anaerobic infections are commonly seen following…

A

trauma or surgery

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12
Q

Two soft tissue infections…

A

-cellulitis
-gas gangrene

other infections include food poisoning, tetanus, Pseudomembranous colitis

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13
Q

What specimens are used?

A

-Body fluids (not urine)
-Deep abscess exudates
-Transtracheal
-specimens/lung aspirates
-Tissue biopsies
-CSF (only in shunt infections)

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14
Q

What are inappropriate specimens?

A

-Sputum, bronchial washings or BAL, gingival
-Gastric, feces, rectal
-Surface material
-Urine
-Vaginal

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15
Q

Anaerobic culture media…

A

-Primary: CDC anaerobic agar, Brucella agar
-Thioglycollate broth
-Chopped meat broth

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16
Q

Selective media

A

-PEA: Phenylethyl Alcohol
-LHV: Laked Kanamycin Vacomycin
-BBE: bacteroides Bile Esculin

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17
Q

Selective media that Inhibits enteric GNR and swarming by some Clostridia

A

PEA: Phenylethyl Alcohol

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18
Q

Selective media that selects for Prevotella and Bacteroides

A

LKV: Laked Kanamycin Vancomycin

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19
Q

What is PRAS media?

A

Pre-reduced Anaerobically Sterilized media

-reduces oxygen presence

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20
Q

Anaerobic incubation at ______ temp.
Incubate _____hours before initial check of plates.

A

35-37

48

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21
Q

Anaerobic incubation finalize culture at _____ days.

A

5-7

Exception—joints may be held for 14 days, for Propionibacterium

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22
Q

anaerobic systems:

Pouch, jars and boxes use CO2 or CO2and _________ generators

A

Hydrogen

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23
Q

Anoxomat II:

Evacuation method- vacuum removes air and then replaces with the following 3 gases…

A

-80% to 90% nitrogen
-5% to 10% hydrogen
-5% to 10% CO2

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24
Q

Anaerobic Glove Box:

5% CO2, 10% hydrogen, and 85% nitrogen, plus a __________ catalyst (reacts with water to produce hydrogen and carbon dioxide), maintain the anaerobic environment inside the chamber.

A

palladium

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25
Q

When subculturing for aerotolerance, what agar is used?

A

Chocolate agar in CO2

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26
Q

After incubation, what is examined?

A

-Colony morphology
-Hemolysis
-Pigments/fluoresce

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27
Q

Aerotolerance test?

A

?

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28
Q

What is Ludwig’s angina?

A

infection of oral cavity

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29
Q

Chopped meat broth:

most use ______ as source of energy.

A

protein

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30
Q

What plate can provide anaerobic conditions?

A

chocolate agar

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31
Q

What percentage of oxygen is in the candle jar method?

A

10%

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32
Q

Identification tests?

A
  1. Catalase
  2. Indole (p-dimethylaminocinnamaldehyde – DMCA)
  3. Special Potency Antibiotic Disks
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33
Q

What are the special potency antibiotic disks? And what is SPS used for

A

-Kanamycin
-Vancomycin
-Colistin
-SPS (for GPC only)

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34
Q

Esculin hydrolysis is used to differentiate…

A

Bacteroides sp. (causes abscess)

-Dark brown or black complex

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35
Q

What is in Bacteroides Bile Esculin Agar (BBE) and what is it used for?

A

-gentamicin and oxgall-inhibit facultative anaerobes and most GN anaerobes

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36
Q

What is a positive result when using Bile Esculin Agar (BBE)?

A

dark brown or black complex

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37
Q

What type of media is Egg Yolk Agar?

A

Nonselective and differential

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38
Q

What is in Egg Yolk Agar?

A

-Lecithinase (Nagler test)
-Lipase
-Proteolytic activity

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39
Q

Egg Yolk Agar:

Degradation of lecithin =

A

opaque precipitate surrounding area of growth

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40
Q

Egg Yolk Agar:

Lipase positive…

A

iridescent sheen on colony surface (breakdown of free fats)

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41
Q

Egg Yolk Agar:

Proteolytic activity=

A

clearing around colony (proteins)

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42
Q

What agar is used for differentiation of Clostridium sp.?

A

Egg Yolk Agar

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43
Q

What is the reverse CAMP test used to differentiate?

A

Clostridium perfringens from other Clostridium sp.

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44
Q

The alpha toxin produced by Clostridium perfringens interacts with CAMP factor from ____________ and produce synergistic hemolysis.

A

Streptococcus agalactiae

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45
Q

What indicates a positive reaction for the reverse CAMP test?

A

hemolysis

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46
Q

What SPS susceptibility test used for?

A

For P. anaerobius (susceptible)
Sodium Polyanetholsulfonate disk

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47
Q

What are commercial ID kits usually used for?

A

Blood and cultures and other sites

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48
Q

An identification system used by many hospitals. It uses spectrophotometry.

A

MALDI-TOF

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49
Q

EXAMINATION OF ANAEROBIC
CULTURES
always use what

A

Chocolate agar in CO2

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50
Q

Anaerobic glove box

A

5% CO2, 10% hydrogen, and
85% nitrogen, plus a palladium
catalyst (reacts with water to
produce hydrogen and carbon
dioxide), maintain the anaerobic
environment inside the chamber.

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51
Q

Anoxomat 2

A

Evacuation method- vacuum removes air and then
replaces with the following 3 gases -80% to 90%
nitrogen, 5% to 10% hydrogen, and 5% to 10% CO2

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52
Q

Anaerobic systems

A

Pouch, jars and boxes use CO2 or
CO2and hydrogen generators

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53
Q

Anaerobic incubation

A

*Incubated at 35-37°C
*Incubate 48 hours before initial check of
plates
*Finalize culture 5-7 days
*Exception—joints may be held for 14
days, for Propionibacterium

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54
Q

Candle jar

A

Flame uses oxygen.
Increased CO2 extinguishes flame

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55
Q

PRAS media

A

Pre-reduced anaerobically sterilized media

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56
Q

Identification systemss are

A

commercial ID kits – usually used for blood
cultures and other sites

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57
Q

Gas liquid chromatography

A
  • Separates components
  • All use mobile phase & stationary (separating )
    phase
  • Anaerobes:
    – Short chain volatile end-products or
    – Fatty acid cellular components
    Gas: mobile phase, Liquid: stationary phase
    Sample: volatized into column
    Acids are recovered in specific order based on
    chemical/physical properties
    Compared to standard
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58
Q

Steps for anaerobic infection

A
  1. Gram stain
  2. Aerotolerance test
  3. Decide which test to set up
    for identification
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59
Q

ID pathway for Gram - rod

A
  • Indole
  • Kanamycin, Colistin,
    Vancomycin disks
  • Bile esculin
  • LK agar, optional
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60
Q

ID pathways for gram + rod

A

Large
*EYA
*Reverse CAMP test,
optional
Small:
*Catalase
*Indole

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61
Q

ID pathway for gram positive cocci

A
  • disk
  • Vancomycin disk
  • Colistin disk
  • Indole
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62
Q

ID testing for gram-negative cocci

A

No further testing

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63
Q

Anaerobic gram postive cocci

A
  • Peptostreptococcus
  • Peptococcus
  • Finegoldia
  • Staphylococcus saccharolyticus
  • Gallicola
  • Parvimonas
  • Peptoniphilus
  • Murdochiella
  • Anaerococcus

*** 9 species and some not studied

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64
Q

Anerobic gram positve cocci

normal microbiota

A
  • Oral cavity, upper respiratory tract, GI tract,
    female genitourinary tract and skin
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65
Q

Anerobic gram positve cocci

Clinical manifestations

A
  • A great variety of infections including abscesses,
    gangrene, cellulitis, bacteremia, pneumonia,
    peritonitis, bite wounds, and pelvic inflammatory
    disease.
  • Brain abscesses: 40% mortality
66
Q

Gram +anaerobes cocci

Difficult to speciate

A

P. anaerobius: susceptible to sodium polyanethol
sulfonate (SPS)
P. asaccharolytic: indole positive
P. micros: very small cocci
Peptococcus niger: black colony
Finegoldia magna: large cocci; look like
staphylococci

67
Q

Peptostreptococcus

A

Anaerobic GPC can form tetrads, chains, clusters, pairs
or clumps and can vary in size of the cocci

68
Q

Gram - cocci

Vellonella

A
  • Veillonella sp. (historically the only one in
    human infections)
    – Part of the normal microbiota-oral cavity,
    genitourinary, respiratory, and GI tract
    – Usually in mixed infections; rarely significant
    – Biochemically inert
69
Q

Gram negaive cocci

other less common genera

A

– Megasphaera
– Anaeroglobus
– Negativicoccus
– Acidaminococcus

70
Q

Anaerobic gram negative rods

A
  • Bacteroides
  • Prevotella
  • Porphyromonas
  • Fusobacterium
71
Q

Baceroides fragilis group are the most of any what

A

· They are the anaerobic bacteria most often
isolated from human infection
· They are the most common of all anaerobes in
producing life-threatening illness

72
Q

Bacteroides fragilis group

All are

A

Catalase positive

73
Q

B fragilis is indole

A

Indole -

74
Q

B.thetaiotamicron is

A

the second most common species

Indole +

75
Q

B.ovatus and B.uniformins is

A

Indole +

76
Q

B.vulgatus and B.distasonis is

A

Indole -

77
Q

Other species of bacteroides are

A

B. eggerrthii, B. merdae, B.
stercoris, and B. caccae.
* B. ureolyticus

78
Q

Bacteroides spp-
beneficial normal flora

Produce

A

butyrate (SCFA) which provides
energy to enterocytes

79
Q

Bacteroides spp-
beneficial normal flora

Play a role in

A

bile acid recirculation and
biotransformation/help ferment carbs

80
Q

Bacteroides spp-
beneficial normal flora

Compete with

A

pathogenic microorganisms

81
Q

Bacteroides spp-
beneficial normal flora

synergy

A

with other coliforms

82
Q

Bacteroides spp-
beneficial normal flora

Note

A

30-50% of fecal matter may be
composed of B. frag

83
Q

Bacteroides are not always beneficial

A

*Capsule confers resistance to host
mechanisms
*Fibrin deposition and abscess formation can
be stimulated by capsular material
*Most common infections are abscesses
which may lead to septicemia

84
Q

diseases of Bacteriodes

A

Intra-abdominal infection, skin & soft
tissue
infection, bacteremia

85
Q

Laboratory diagnosis and culture of Bacteroides

A

Gram stain: GNR with rounded ends,
safety pin appearance (containing vacuoles)
Culture: B. fragilis exhibit
Concentric whirls or ring-like structures
(under stereoscope)

86
Q

ID tests of Bacteroides

A

· Growth in the presence of 20% bile: positive
· Penicillin: resistant
· Kanamycin: resistant
· Vancomycin: resistant
· Colistin resistant

87
Q

Prevotella and porphyromonas genera

they are the 2nd most

A

common group of anaerobic bacteria

88
Q

Prevotella and porphyromonas genera

they are normal inhabitant of the

A

oropharynx,
nose, GIT, GUT

89
Q

Prevotella and porphyromonas genera

most of them develop

A

pigmented colonies
(brown or black) and demonstrate florescence
under UV light (brick red)

90
Q

Prevotella and porphyromonas genera

They are mostly associated with what infection

A

orofacial and
pleuropulmonary infections

91
Q

Fusobacterium genus

important member

A
  • F. nucleatum
  • F. necrophorum
  • F. mortiferum
  • F. varium
92
Q

Fusobacterium genus

diseases

A

pleuropulmonary infection, intra-abdominal
infection, abscess, systemic infection , Vincent’s disease
(anaerobic tonsillitis)

93
Q

Fusobacterium genus

gram stain

A

slender, spindle-shaped
GNR with tapered end
Others – pleomorphic GNR with spherical swelling

94
Q

Fusobacterium genus

culture

A

· F. nucleatum: speckled opalescent appearance,
bread-crumb colonies
· F. necrophorum: convex with irregular margins
· F. mortiferum, varium: fried egg appearance

95
Q

ID test of Fusobacterium

A

-Fusobacterium sp. are susceptible to
Kanamycin and colistin-
no growth on KV BAP
-F. mortiferum is positive for esculin
hydrolysis and lactose fermentation,
-F. varium is negative for both
GLC: butyric acid peak

96
Q

Vincents disease description in detail

A

· It is a necrotizing gingivostomatitis
·It is caused by Treponemes, P.
melanogenica,
other anaerobic GNR and anaerobic cocci
·Fusobacterium necrophorum is involved
with the severe form of the disease, which is
accompanied by sepsis

97
Q

Vancomycin(5 ug)

A
  • Majority of Gram positive organisms are
    susceptible
  • Exception-some species of Lactobacillus sp.
  • Majority of Gram negative organisms are
    resistant
  • Useful when organism is Gram variable or falsely
    staining negative (some Clostridia)
98
Q

kanamycin

A

1000ug

99
Q

Colistin

A

10ug
Most gram postive are resistant

100
Q

Prevotella

disk suceptible or R and testing

A

Flourescence= +
Pigment= may be + or -
BBE= negative
Kanamycin=R
Vanvomycin=R
Colistin= R or S
Growth on LKV= growth
Glucose fermentation= +

101
Q

Porphyromonas

Disk su or Re and testing

A

Fluorescence +
Pigmentation +
BBE= neg
Kanamycin= R/S
Vancomycin= S
Colistin= R
growth on LKV= negative
glucose fermentation= negative

102
Q

B.fragilis group resistance or susceptibility

A

Fluorescence -
Pigmentation -
BBE= growth
Kanamycin= R
Vancomycin= R
Colistin= R
growth on LKV= growth
glucose fermentation= Postive

103
Q

Fusobacterium

A

Fluorescence -
Pigmentation -
BBE= neg
Kanamycin= S
Vancomycin= R
Colistin= S
growth on LKV= negative
glucose fermentation= NA

104
Q

Anaerobic spore-forming rods

A

clostridium, botulism, Tetani, perfringes, difficile

105
Q

4 types of botulism

A
  1. Classical or food-borne: ingestion of improperly canned food
  2. Infant botulism: ingestion of botulinal spores from soil, dust, honey
  3. Wound botulism: contamination of traumatic wound with botulinal spores
  4. Undetermined: unknown source
106
Q

C.Botulinum symptoms

A

weakness, dizziness, blurred vision, muscle weakness,
respiratory and flaccid paralysis

107
Q

C.Botulinum virulence factors

A

Exotoxin that blocks the release of acetylcholine

108
Q

C.botulinum Lab diagnosis

A

Demonstration of C. botulinum and/or its toxin
in specimens

109
Q

Clostrium tetani

disease

A

Tetanus

110
Q

Clostridium tetani

symptoms

A

muscular spasm, lockjaw, respiratory distress

111
Q

C.Tetani virulence factors

A

tetanospasmin (neurotoxin) that
inhibits inhibitory neurotransmitters of motor neurons

112
Q

C.Tetani diagnosis

A

it is usually based on
clinical symptoms

113
Q

C.tetani gram stain and what is availble

A

drumstick or tennis-racket
appearing GPR
Vaccine available

114
Q

Clostridium perfringens diseases

A

(Soft tissue):
1. Wounds
2. Cellulitis
3. Myonecrosis or gas gangrene: caused by 12 toxins
(lecithinase or a-toxin)
4. Food poisoning: caused by enterotoxin

115
Q

Gas gangrene may be caused by

A

other Clostridia like C.
novyi, C. septicum, C. sordellii, C. histolyticum

116
Q

Gas gangrene most common cause

A

C. perfringens is the most common agent

117
Q

Laboratory diagnosis of Clostridium per-fringes

Gram stain and culture

A

Gram stain: large, box-car shaped spore-forming
GPB
Culture: double zone of b hemolysis

118
Q

Laboratory diagnosis of Clostridium per-fringes

ID tests

A

· Positive Nagler test (lecithinase test): opaque
precipitation
· Positive reverse CAMP tes

119
Q

Reversed CAMP test

A

The alpha toxin
produced by
Clostridium
perfringens interacts
with CAMP factor
produced by Group B
Streptococcus and
produce synergistic
hemolysis.

120
Q

C.difficile is a

A

Gram postive spore forming bacilli

121
Q

C difficile diseases

A

Antibiotic associated diarrhea or
colitis, pseudomembranous colitis

122
Q

C difficile

antibiotic therapy

A

Antibiotic therapy will diminish the normal
bowel flora, which may be followed by
intestinal colonization with Clostridioides
difficile

123
Q

C.difficile

CDC facts

A
  • It’s estimated to cause almost half a million
    illnesses in the United States each year.
  • About 1 in 5 patients who get C. diff will get
    it again.
  • Within a month of diagnosis, 1 in 11 people
    over age 65 died of a healthcare-associated
    C. diff infection.
124
Q

C.difficile risk factors

A
  • Age - elderly
  • Hospitalizations or long-term care
  • Antibiotic treatment
  • Previous infection
  • Recent exposure to organism
125
Q

C.difficile colonization

0-6 months

A

0-37% carriage rates

126
Q

C.difficile colonization

6-12 months

A

14% carriage rates

127
Q

C.difficile colonization

by 3 years of age

A

~3% = non-
hospitalized adults

128
Q

C.difficile testing is not

A
  • Testing is not recommended in <1
    years of age and other causes should be
    ruled out before testing in ages 1-3
    years
129
Q

C.difficile testing

many PCR tests

A
  • Many PCR tests are not validated in
    children 2 years of age and younger
130
Q

C.difficile symptoms

A
  • Loose (watery) stools
  • Fever
  • Tender stomach or pain
  • Fever
  • Loss of appetite
  • Nausea
131
Q

C.difficile complications

A
  • Pseudomembranous Colitis
  • Recurrence – up to 30% of patients will
  • relapse with the same strain or a new strain
132
Q

C.difficile virulence factors

A

Toxin A, toxin B test by
NAP1/B1/027 (North American pulsed-field gel EF type
1, RE type B1, PCR ribotype 027) – For hypervirulent
strain

133
Q

C.difficile Lab diagnosis

tissue culture cytotoxin assay

A

demonstration of toxin B

134
Q

C.diff Lab diagnosis

Culture isolation agar type

A

Cycloserine-cefoxitin egg yolk fructose
agar (CCFA)

135
Q

C.diff Lab Diagnosis

GDH

A

(Glutamate dehydrogenase

136
Q

C.diff Lab Diagnosis

toxin testing

A

(EIA)

137
Q

C.diff Lab Diagnosis

PCR for

A

toxin A and or B gene detection

138
Q

C.diff Lab Diagnosis

specificity and sensitivity in

A

the high 90-100% range depending on the manufacturer

139
Q

C.diff Lab Diagnosis
detecting gene does not

A

Mean it is producing the gene

140
Q

CCFA

A

(Cycloserine-cefoxitin egg yolk
fructose)

141
Q

Cycloserine-cefoxitin egg yolk
fructose

A
  • Enriched, selective, and differential
  • Consists of animal peptones and fructose, and
    supplemented with cefoxitin and cycloserine at to
    inhibit growth of most normal fecal flora.
  • Cycloserine -inhibits Gram-negative bacteria
  • Cefoxitin will inhibit both Gram-positive and Gram-
    negative organisms
142
Q

Cycloserine-cefoxitin egg yolk
fructose.

C.difficile is not

makes characteristic

turns neutral PH

A
  • C. difficile is not inhibited
  • Makes characteristic yellow, ground-glass colonial
    morphology.
  • It turns the neutral red pH indicator from pink/orange
    to yellow (change in pH from utilization of the amino
    acids)
143
Q

C.difficile algorithm

A

One testing algorithm

A One test algorithm may be used if the bacteria is GDH positive and Toxin positive then the final report is positive. If the bacteria are GDH positive and Toxin negative, then a Genexpert is used to determine if the organism is positive. If both are negative, then the clinical report is negative

144
Q

Treatment for C.difficile

A
  • Metronidazole
  • Vancomycin
  • Severe or Recurrent infection:
  • Both metronidazole and vancomycin
  • Fidaxomicin
  • FMT (fecal microbiota transplant)
  • Monoclonal antibodies
145
Q

Transmission/prevention for C.difficile

A
  • Fecal oral route
  • Spread by spores
  • Spores can survive on surfaces for months to
    years
  • Frequent HAI (Hospital Acquired Infection)
  • Patients must be isolated (contact isolation)
  • Bleach or other sporicidal cleaners must be
    used
146
Q

Anaerobic non-spore-forming Bacilli
Members:

A

· Propionibacterium
· Eubacterium
· Actinomyces
· Bifidobacterium

147
Q

Propionibacterium acnes

Is responsible for

A

Human acne

148
Q

Propionibacterium acnes

can be found as

A

· It can be found as normal flora in the mouth, skin, GIT, GUT,
nasopharynx

149
Q

Propionibacterium acnes

It is a frequent

A

· It is a frequent contaminant of blood cultures

150
Q

Propionibacterium acnes

Can cause

A

· Can cause joint infections and CSF shunt infections

151
Q

Propionibacterium acnes

gram stain and culture

A

· Gram stain: diphtheroid-like or club-shaped GPR with Chinese
letter formation
· Catalase positive and indole positive

152
Q

Eubacterium lentum ( now Eggerthella lenta)

A

· It is a true agent of endocarditis (normal oral
flora)
· It is recovered from mixed culture from wound
or abscess
· Gram stain: diphtheroid-like GPR

153
Q

Actinomyces species

disease and infection is initiated

A
  • Disease: actinomycosis (cervicofacial or lumpy jaw, thoracic,
    abdominal)
    Note: Infection is initiated when the endogenous organism is inoculated into damaged tissue
    after another infection, trauma, or surgical manipulation
154
Q

Actinomyces species symptoms

A

formation of draining sinus tract containing “sulfur
granules”

155
Q

Actinomyces species Lab Diagnosis

Gram stain and culture

A
  • Gram stain: Branching, beaded Gram-positive rods
  • Culture: CDC-BAP young colonies of A. israelii look like spider
    colonies (slow-grower)
156
Q

Actinomyces species Lab Diagnosis

older colonies

A
  • Older colonies are referred to as molar tooth (heaped or raised up)
157
Q

Actinomyces species Lab Diagnosis

The most Frequent species

A

The most frequent species involved in actinomycosis is
Actinomyces israelii

158
Q

Bifidobacterium dentium

Inhabitant and gram stain

A
  • It is a normal inhabitant of the oral cavity and
    the GI tract
  • Gram stain: Y or bifid shaped Gram-positive
    rods
159
Q

Bifidobacterium dentium

Are what

A
  • “Friendly” Probiotic Along with
    Lactobacillus,
  • inhibits harmful microorganisms
  • (Claimed to promote good digestion, boost the
  • immune system.)
160
Q

Mobiluncus

A
  • Strict anaerobe
  • Gram variable curved rods with tapered
    ends
  • Normal flora but believed to be involved in
    Bacterial Vaginosis.
  • Usually not cultured in Clinical lab because
    found in samples unacceptable for
    anaerobic culture

Medical Microbiology II - 4336 (20 decks)

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