Neisserieae & Haemophilus

Front 1

Neisseriaceae includes:

Back 1

Neisseria
Moraxella
Kingella
Acinetobacter

Front 2

Fitz-Hugh-Curtis syndrome ?

Back 2

Ascending infection caused by GC
---spreading of organism from fallopian tubes to liver and peritoneum

Front 3

GC is extremely sensitive to____?

Back 3

Drying and extreme temperatures

DO NOT REFRIGERATE!!!

Front 4

GC specimens should include:

Back 4

vagina/urethra, oropharynx, rectum

Front 5

GC specimens can be kept at room temperature for how many hours?

Back 5

up to 6 hours

Front 6

Growth Requirements for GC?

Back 6

35-370C
Capnophilic (5-7% CO2)

Front 7

Male specimens - urethral discharge
gram stain --Positive for GC ?

Back 7

intra and/or extracellular gndc and PMNs

Front 8

Gram stain Female specimens – endocervical swabs
presumptive GC?

Back 8

presumptive GC = >3 neutrophils/ hpf with intracellular gndc
50-70% sensitive/ evaluate with symptoms
Should be always be confirmed by culture

Front 9

Cystine Trypticase Agar (CTA)?
Confirmatory Identification Tests for Neisseria species

Back 9

Semisolid agar + 1% gluc., malt., lact., or suc.
Phenol Red = pH indicator used to detect acid
Indicator changes from Red(alkaline) to yellow (acid)
1% of sugar + phenol red + acid = yellow

Front 10

Confirmatory Identification Tests for Neisseria species
Rapid Acid detection tests?

Back 10

Use bacterial enzymes to detect acid production from carbohydrates
Results between 2-4hrs

Front 11

Neisseria meningitidis causes which two major types of infections?

Back 11

Meningitis (a.k.a Meningococcal meningitis)
Septicemia (a.k.a Meningococcemia)

Front 12

the 4 most important N. meningitidis Groupes that based on its polysaccharide capsule?

Back 12

A, B, C, Y and W135

Front 13

Classic signs and symptoms have sudden onset of Meningococcal meningitis?

Back 13

High fever
Headaches
Nuchal rigidity
Flu-like symptoms
Confusion
Vomiting in young children

Front 14

Meningococcal meningitis
Patients exhibit --- Waterhouse- Friderichsen Syndrome ?

Back 14

Acute hemorrhagic necrosis of adrenal glands

Front 15

secondary prevention programs

Back 15

Identifies persons in the early stages of problem behaviors and ... does not have to be the case

Front 16

N.menigitis is BIOSAFETY LEVEL 2 ORGANISM, correct?

Back 16

yes

Front 17

gram stain of N. meningitidis?

Back 17

1.Gram-negative diplococci + PMNs
2.CSF cell count >1,000 PMNs/μL
3.CSF glucose ↓ protein ↑

Front 18

Serological Assays for N. meningitidis?

Back 18

1.Latex Agglutination Test
2.Direct antigen test that detects meningococcal capsular polysaccharide A, B,C, Y, and W135
---Specimen used: CSF, urine, and serum
----Should be used in conjunction with culture and gram stain

Front 19

Media and Growth Requirements for N. meningitidis?

Back 19

1.CHOC and BAP
2.Selective media for nasopharyngeal swabs (MTM, ML)
3.Aerobic, 35-370C , 5-7% CO2, 24/48/72hrs

Front 20

Identification Tests for N.meningitidis?

Back 20

BAP = GROWTH
CHOC = ML= GROWTH
Gram Stain – GNDC
Oxidase test - POSITIVE
Catalase – 3% H2O2 - POSITIVE
Superoxol – 30% H2O2 –WEAK POSITIVE OR NEGATIVE
Colistin disk – resistant
Acid Production from Glucose and Maltose
Enzyme detected γ-glutamyl aminopeptidase

Front 21

saprophytic Neisseria (other Neisseria species)?

Back 21

1.NF in the Upper Respiratory Tract
2.Opportunistic pathogen in human
3.Grows on BAP; most do not grow on ML/MTM
4.Not routinely identified in labs; usu. considered a contaminant

Front 22

Identification Tests for common Neisseria species (N. cinerea)?

Back 22

Easily misidentified as GC
BAP = GROWTH*
CHOC = GROWTH
ML/MTM = GROWTH(Variable)
Superoxol = NEGATIVE*
Acid Production from Glucose(weak)
Enzyme detected= hydroxypropyl-aminopeptidase
Colistin disk-SENSITIVE*

Front 23

Identification Tests for common Neisseria species
N.lactamica?

Back 23

BAP = GROWTH (YELLOWISH COLONY)
CHOC = GROWTH
ML/MTM = GROWTH
Superoxol = NEGATIVE
Acid Production from Glucose, Maltose, Lactose

Front 24

Identification Tests for common Neisseria species
N. sicca?

Back 24

BAP = GROWTH
CHOC = GROWTH
ML/MTM =NO GROWTH
Acid Production from Glucose, Maltose, Sucrose

Front 25

Hint to remember Acid Production from Carbohydrates

Back 25

Glucose = Gonorrhea
Maltose = Meningitidis
Lactose = Lactamica
Sucrose = Sicca or Species

Front 26

Drug of choice for GC?

Back 26

Ceftriaxone (also cefixime, ciprofloxacin, ofloxacin)
In combo with doxycycline or erythromycin
Silver nitrate or penicillin drops/ointment-eye infections

Front 27

drug of choice for N.meningitidis?

Back 27

penicillin G

Front 28

prevention of N. meningitidis?

Back 28

vaccine
1.Available for serogroups A, C, Y, and W135
2.Group B available but not licensed in U.S.
3.Offers up to 3 yrs protection
4. Not effective < 2 yrs children

Front 29

Clinical Significance of Moraxella catarrhalis?

Back 29

1.Commonly found in URT of children (~51%)
2.Majority of clinical diseases include otitis media, pneumonia, sinusitis, bronchitis, conjunctivitis

Front 30

Moraxella catarrhalis
Gram-stain of sputum:

Back 30

Many PMNS and GNDC >25 under 100X

Front 31

Moraxella catarrhalis
Colony Morphology?

Back 31

1.Grows on BAP and CHOC; some strains grow well on MTM
2.No growth on MAC
3.Smooth, opaque, white-gray slides on agar plate =Hockey-Puck

Front 32

identification tests for Moraxella catarrhalis?

Back 32

1.Gram stain = GNDC
2.Oxidase = POSITIVE
3.Catalase = POSITIVE
4.Asaccharolytic = does not utilize sugar

5.DNase = POSITIVE
6. Bcat disk (Indoxyl butyrate hydrolysis spot test) POSITIVE

Front 33

Haemophilus species inhabit respiratory tract, but ______
always a pathogen?

Back 33

H. ducreyi

Front 34

Haemophilus influenza:
Most common encountered strains in disease is ____?

Back 34

type b

Front 35

H. influenzae has which two broad categories?

Back 35

Encapsulated strains (typeable)
Non-Encapsulated strains (non-typeable)

Front 36

H. influenzae can cause what kind the disease in children?

Back 36

Meningitis
--Occurs mostly in children (2 mos-3yrs)
Epiglottitis
Pneumonia

Front 37

which organism assoc. with severe purulent conjunctivitis “pink eye” and Brazilian Purpuric Fever (BPF)

Back 37

H. influenzae biogroup aegyptius

Front 38

HACEK represent which organisms?

Back 38

Haemophilus (aphrophilus & paraphrophilus)
Actinobacillus
Cardiobacterium
Eikenella
Kingella

Front 39

H. paraphrophilus?

Back 39

Found in NF of human
Can cause endocarditis

Front 40

which two hemophilus organisms could found in human dental plaque?

Back 40

H. parahaemolyticus
H. segnis

Front 41

Gram stain Hemophilus species?

Back 41

Small gram negative pleomorphic rods, coccobacillary to filamentous
Often tiny and faint staining

Front 42

Hemophilus species is motility positive or negative?

Back 42

negative

Front 43

General Characteristics of Hemophilus species?

Back 43

1.Optimal temperature 35-37°C .Exception: H. ducreyi - should be incubated at 33-35°C
2.Capnophilic
Moist environment with ↑ CO2
(5-10%)
3.Colonial morphology
Translucent, dew-like
Very distinctive odor, “mouse nest”, bleach like or musty odor
4.Require X and V growth factors in culture

Front 44

"blood loving" is talking about which organism species?

Back 44

Haemophilus = “blood loving”

Front 45

X and V factors

Back 45

X factor:
A.k.a heme, hematin, hemin
Heat stable
Present in RBC’s

V factor:
"V” for vitamin
A.k.a NAD or satelliting factor
Heat labile
Present in lysed RBC’s

Front 46

X and V factors requirementsTesting Methods:X and V strips/discs

Back 46

Streak basal media for confluent growth
Mueller-Hinton or TSA
Contain neither X nor V factor
Supply factors via discs/strips
X, V, XV discs or strips placed close together
Incubate @350C room air 18-24hrs
Observe growth around discs/strips

Front 47

Result Interpretation for discs/strips

Back 47

Org. requires X factor if:
X and XV disc/strip = growth
V disc/strip = no growth
Org. requires V factor if:
V and XV disc/strip = growth
X disc/strip = no growth
Org. requires X and V factors if:
XV disc/strip = growth
X and V disc/strip = no growth

Front 48

principle of alpha-Aminolevulinic Acid (ALA) or Porphyrin substrate Test?

Back 48

1.Detects the presence of enzymes that convert ALA to porphobilinogen or porphyrins through the heme synthesis pathway
2 Determines the org. X factor requirement
Org. that lack enzymesRequire X factor (Heme)

Front 49

result of ALA test?

Back 49

Wood’s lamp (UV lamp)
Positive = red fluorescent
Negative = clear
Kovac’s reagent
Positive = red/pink color in the lower aqueous phase lower
Negative = no color change lower aqueous phase
NOTE: UV can only be used prior to Kovac’s addition
Positive test= org. possess enzymes that does NOT require X factor

Front 50

what hemophilus type doesn’t need X & V factors (grows on BAP)?

Back 50

H. aphrophilus

Front 51

which two organism need V factors only?

Back 51

H. parainfluenzae and H. segnis

Front 52

organism require X factor only?

Back 52

H. ducreyi

Front 53

H.hemolyticus and H.parahemolyticus both create beta hemolysis on horse blood agar, what is the requirement difference between them?

Back 53

H. hemolyticus – X & V factors w/ β hemolysis on horse blood agar
H. parahemolyticus – V factor w/ β hemolysis on horse blood agar

Front 54

Culture Characteristics of haemophilus?

Back 54

Growth on CHOC
Gram stain=GNCB
V factor requirement
Satelliting on BAP
X factor requirement
ALA
Discs/strips
Quad plate
ß-lactamase testing
-----Nitrocefin disc

Front 55

Haemophilus ducreyi can cause which sexally transmitted disease?

Back 55

“Chancroid”
----Starts as a small painful lesion on or around the genital area
Progress from tender papules to painful ulcers with several satellite lesions
Do NOT cause systemic dz

Front 56

ID of Haemophilus ducreyi?

Back 56

Colonies are “sticky”: can’t be picked up by loop
Secondary GS = gnr “school of fish appearance”
Oxidase = + (can take 15-20sec)
Catalase = negative
SPS disc = S (sodium polyanethol sulfanate)
Cannot use X and V discs nor ALA
Other methods to id
Rapid biochemicals:
Molecular: PCR (culture & direct specs.), DNA homology