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CHAPTER 17

Neisseria species
and
Moraxella catarrhalis
Neisseria spp. and
M.catarrhalis
FAMILIES

Neisseriaceae Moraxellaceae
▪ Neisseria ▪ Moraxella
▪ Kingella ▪ Acinetobacter
▪ Eikenella
▪ Simonsiella
▪ Alysiella
▪ Several other
genera

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GENERAL
CHARACTERISTICS:
Neisseria spp.

▪ Gram negative diplococci


(gndc)
▪ Aerobic
▪ Oxidase positive
▪ Fastidious; capnophilic
▪ Catalase positive
▪ Nonmotile
▪ Nonsporeforming

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GENERAL
CHARACTERISTICS:
Neisseria spp.
Habitat:
▪ Upper respiratory tract (URT)
▪ Genitourinary tract

Primary Pathogens:
▪ N.gonorrhoeae
▪ N.meningitidis

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VIRULENCE
FACTORS:
Neisseria spp.
▪ Capsule
▫ N. meningitidis
▫ anti-phagocytic
▪ Pili/fimbriae
▫ Attachment to host tissue
▪ Lipoligopolysaccharide
(LOS)
▫ Endotoxin
▫ Elicits inflammatory
response
▫ Causes damage to tissues
5 ▪ Protein III
GENERAL
CHARACTERISTICS:
N.gonorrhoeae
▪ Pyogenic infection of columnar and transitional
epithelial cells
▫ Urethra
▫ Endocervix
▫ Anal canal
▫ Pharynx
▫ Conjunctiva
▪ Acquired through sexual contact
▪ Incubation period is 2-7 days
▪ Affects both males and females but presents
differently
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CLINICAL INFECTIONS:
N.gonorrhoeae

Disease in the Male Disease in the Female


▪ 90% cases are symptomatic ▪ 50% cases are
▫ Purulent discharge symptomatic
▫ dysuria ▫ Dysuria
▪ Complication from ▫ Cervical discharge
untreated infection ▫ Lower abdominal pain
▫ Prostatitis ▪ Complication from
▫ epididymitis untreated infection
▫ Pelvic inflammatory
disease (PID)
▫ Sterility
▫ Ectopic pregnancy

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CLINICAL INFECTIONS:
N.gonorrhoeae
Disseminated gonococcal disease
▪ Bloodborne dissemination occurs in <1% of all infections

▪ Symptoms:
▫ Fever
▫ Rash
▫ Bacteremia

▪ Possible result of bacteremia:


▫ Gonococcal/septic arthritis
▫ septic joints
▫ Swollen
▫ pus/synovial fluid filled

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CLINICAL INFECTIONS:
N.gonorrhoeae
IN INFANCY
▪ Newborns may acquire eye infection, ophthalmia
neonatorum, during vaginal delivery through infected birth
canal

▪ May result in blindness

▪ Infection prevented with antibiotic (erythromycin) eye drops


applied at birth

EXTRAGENITAL INFECTIONS
▪ Pharyngitis
▪ Anorectal infections
▪ Ocular infections
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LABORATORY DIAGNOSIS:
N.gonorrhoeae
Clinical Specimens
▪ Genital sites
▫ FEMALE: endocervical
▫ MALE: urethral
▪ Anorectal
▪ Oropharyngeal
▪ Eye
▪ Blood/synovial fluid

Collection and Transport


▪ Dacron/rayon swabs preferred
▪ Transport in Amies with charcoal
▪ Inoculate media within 6 hours of collection
▪ Direct plating to media also possible
▫ JEMBEC
▫ TransGrow
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LABORATORY DIAGNOSIS:
N.gonorrhoeae
Gram stain morphology
▪ Gram negative
▪ Diplococci
▪ kidney/coffee bean shaped

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LABORATORY DIAGNOSIS:
N.gonorrhoeae
Media selection, Incubation
▪ Chocolate agar
▫ Many OTHER organisms can also grow
▫ Potential overgrowth of normal flora
▪ Thayer-Martin (TM) agar
▫ Chocolate agar PLUS
▫ Vancomycin to inhibit gram positive organisms
▫ Colistin to inhibit gram negative organisms
▫ Nystatin to suppress the growth of yeast
▪ Modified Thayer-Martin (MTM) agar
▫ Same as Thayer-Martin PLUS
▫ Trimethoprim to prevent swarming by Proteus sp.
▪ Specimens held at room temp, plated ASAP
▪ Media should be pre-warmed to avoid cold shock
▪ Incubate at 35°C, 3-5% CO2, held for 72 hours
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LABORATORY DIAGNOSIS:
N.gonorrhoeae
Colony morphology
▪ On Choc/TM/MTM
▫ Small
▫ Grey-tan
▫ Translucent
▫ Raised
▪ Autolytic enzymes; need fresh
growth
Oxidase Test
▪ Oxidase disc + organism
▪ POSITIVE: dark blue/purple color
▪ NEGATIVE: no dark blue/purple

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LABORATORY DIAGNOSIS:
N.gonorrhoeae
Carbohydrate Utilization
▪ Cystine trypticase agar (CTA)
▫ 1% of a single carbohydrate
▫ glucose/dextrose
▫ maltose
▫ lactose
▫ sucrose
▫ Phenol red indicator to detect acid
▫ Read in 24-72 hours
Glucose Maltose Lactose Sucrose
(dextrose)

N.gonorrhoeae + = = =

N.meningitidis + + = =

N.lactamica + + + =

14 N.sicca/subflava + + = +
LABORATORY DIAGNOSIS:
N.gonorrhoeae
Identification Methods
▪ Chromogenic/multi-test biochemical
▫ Detect enzymes that produce colored end products
▫ Combine enzyme substrate test with traditional
biochemical tests
▪ Immunologic
▫ Does not require pure or viable (living) organisms
▫ Examples are coagglutination and fluorescent
antibody testing
▪ Nucleic acid amplification tests (NAATs)
▫ Preferred assay with increased sensitivity/specificity
▫ Noninvasive urine specimen
▪ MALDI-TOF MS
▫ Emerging technology
▫ Identifies unique proteins signatures like mass spec.
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LABORATORY DIAGNOSIS:
N.gonorrhoeae
Antimicrobial resistance
▪ Drug of choice was penicillin
▪ In 1976, 1st penicillinase-producing N.gonorrhoeae (PPNG)
strains isolated
▪ Multiple other resistance patterns since then
▪ CDC developed monitoring system

▪ TREATMENT: Current recommendation is cephalosporin and


cross-coverage for chlamydia since many are co-infected
▪ LAB TESTING: Beta lactamase testing is still performed on all
isolates and the results reported
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N.meningitidis

▪ Found only in humans
▪ Normal flora in the nasopharynx of
carriers
▪ Transmitted by respiratory droplets
from carrier to host
▪ Only a few of those newly colonized
develop disease
▫ Infants
▫ Adolescents
▪ Bacteria may enter the bloodstream
and/or the CNS
▫ Meningococcal septicemia or
Meningococcemia
17 ▫ Meningitis
VIRULENCE FACTORS:


N.meningitidis
▪ Capsule
▫ N. meningitidis
▫ anti-phagocytic
▪ Pili/fimbrae
▫ Attachment to host tissue
▪ Lipoligopolysaccharide
(LOS)
▫ Endotoxin
▫ Elicits inflammatory
response
▫ Causes damage to tissues
18 ▪ Protein III
CLINICAL INFECTIONS:


N.meningitidis
▪ Risk factors for invasive disease
▫ Asplenia
▫ C’ deficiencies
▫ Smoking/exposure to smoke
▫ URT viral infection
▫ Crowded living conditions
facilitates spread
▫ Military barracks
▫ College dormitories
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CLINICAL INFECTIONS:


N.meningitidis
▪ Bacterial meningitis
▫ Transmitted by respiratory droplets
▫ Requires close contact
▫ Symptoms:
▫ Frontal headache
▫ Stiff neck
▫ photophobia
▫ Serotypes B and and C most common in the
US
▪ Other infections
▫ meningococcemia
▫ arthritis
▫ pericarditis
▫ pneumoniae
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CLINICAL INFECTIONS:


N.meningitidis
▪ Meningococcemia
▫ 25% mortality even if treated
▫ Symptoms:
▫ purpura
▫ Petechial skin rash
▫ Tachycardia
▫ Hypotension
▫ thrombosis
▫ Worst case:
▫ DIC
▫ Septic shock
▫ Waterhouse-Friderichsen syndrome
▫ Death is possible in 12-48 hours from onset

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LABORATORY DIAGNOSIS:


N.meningitidis
▪ Gram stain
▫ CSF direct smear; cytocentrifugation
recommended
▫ Examine for intra/extra-cellular gndc
▫ Clearing around organisms may
suggest capsule
▪ Media selection, incubation
▫ Sheep’s Blood agar (SBA),
Chocolate (CA) and/or selective
medium, depending on source
▫ Specimens held at room temp,
plated ASAP
▫ Media should be pre-warmed to
avoid cold shock
▫ Incubate at 35°C, 3-5% CO2, held for
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72 hours
LABORATORY DIAGNOSIS:


N.meningitidis
▪ Colony morphology
▫ Growth on BAP, CA
▫ Small, grey, convex colonies.
Mucoid if encapsulated.
▪ Oxidase test
▫ Oxidase disc + organism
▫ POS= dark blue/purple color
▫ NEG= no dark blue/purple
▪ Carbohydrate utilization
▫ Glucose and maltose acid
production (yellow) = POS
▪ Serologic grouping

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TREATMENT/PREVENTION:


N.meningitidis
▪ Treatment
▫ Penicillin for meningitis
▫ 3rd gen cephalosporins for
meningococcemia
▫ Prophylaxis for exposure with
rifampin or ciprofloxacin

▪ Meningococcal vaccine
▫ Covers some serotypes, not
ALL
▫ Recommended at 11/12 y with
booster at 16 y

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NONPATHOGENIC
Neisseria spp.




Normal flora of the URT
Occasional flora of the genital tract
Rarely cause disease
▪ Must be able to separate these from
the pathogenic Neisseria.
▪ Some members:
▫ N. cinerea
▫ N. lactamica
▫ N. mucosa
▫ N. sicca/ N. subflava

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Moraxella catarrhalis
▪ Isolated only from humans
▪ Normal flora of the URT
▪ Opportunistic pathogen
▪ Predisposing factors:
▫ Advanced age
▫ immunodeficiency/neutropenia
▫ Chronic disease
▪ Clinical infections:
▫ Pneumonia
▫ Otitis media/sinusitis (3rd most common cause in children)

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LABORATORY
DIAGNOSIS:
M.catarrhalis
Gram stain morphology
▫ Gndc, may be
extra/intracellular
Colony morphology
▫ Grey-white
▫ smooth
▫ “Scoot” when pushed
▫ Colonies turn pink with age
▪ Autolytic enzymes; need fresh
growth
Oxidase Test POS
CTA sugars, all NEG
Beta-lactamase POS
Catarrhalis disc POS

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Neisseria sp. and
Moraxella

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REFERENCES
● Engelkirk, P., & Duben-Engelkirk, J. (2008). Laboratory Diagnosis
of Infectious Diseases: Essentials of Diagnostic Microbiology .
Baltimore, MD: Lippincott Williams and Wilkins.
● http://www.awinhospitalproducts.com/product/35-collection-swab-
amies-medium-w-charcoal-wwcsam-3450
● https://new.fishersci.com/ecomm/servlet/fsproductdetail_10652_6
06366_29104_-1_0
● https://picasaweb.google.com/pia8628/0411microlab#5596104725
803822690/
● Mahon, C. R., Lehman, D. C., & Manuselis, G. (2019). Textbook of
Diagnostic Microbiology (6th ed.). Maryland Heights, MO:
Saunders.

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