Neisseria

Conf. Univ. Dr. Irina Codita

 I. General presentation
• Minidefinition:
– Gram negative cocci, coffee bean-shaped
arranged in pairs, oxidase positive, responsible
for meningococcal disease and sexually
transmitted disease gonorrhoea

https://www.google.ro/ habitat-and-morphology-of-neisseria-gonorrhoeae
 Common symptoms:
-High fever
-Fotophobia
-Neck stiffness
- Confusion,
- Convulsions
- Headache, vomit

Severe outcome,
outcome even with correct,
timely diagnostic:
-Mortalitaty 5-10% after 24-48 h
- 10 -20% sequeles:
Bacterial meningitis -Brain
-Deafness
-Mental
Exantema in a meningitis case
Gonorrhoea

Gonococcal ophtalmitis
Neisseria Gram stain
II. Taxonomy and classification

Bergey Manual 2004:

Domain: Bacteria
Phylum: Proteobacteria
Class: Beta Proteobacteria
Order: Neisseriales
Family: Neisseriaceae
Genus Neisseria

. after Albert Neisser, who discovered the etiological

Neisseria, named
agent of gonorrhea in the pus of patients in 1889.
 Main species

Of medical importance: fastidious species

• Neisseria meningitidis
• Neisseria gonorrhoeae

Non-pathogenic Neisseria: non-fastidious

29 species - normally colonize mucosal surfaces of
oropharynx and nasopharynx and occasionally
anogenital mucosal membranes
 III. Habitat
– Neisseria gonorrhoeae is an obligate parasite of
humans – special afinity for urogenital tract (fimbria,
proteic adhesines), may colonise other mucous and
serous surfaces: eye conjunctiva, vagina, rectum
– Neisseria meningitidis occurs in healthy carriers
and may cause epidemics of meningitis.
Carriage rate in general population: 5-30% (nose and throat
of carriers and ill people; other colonisation sites:
conjunctiva, vagina, urethra, rectum);
Carriage rate in closed communities: up to 80% in epidemic
context (e.g. military)
– Other non-fastidious Neisseria species may be
opportunistic pathogens
Species of medical importance . Common traits
 Gram negative, 0,6 – 1m, coffee beans shaped cocci,
adjacent sides flattened
• In pairs (diplo)
• Oxidase positive (cytochrome oxidase)
• Catalase positive
• Fastidious
- Sensitivity to temperature variations (prefere 37 0 C)
- Need special media with different nutritive supplements:
organic (blood, serum etc.) or chemical defined (vitamins,
nucleotides, amino acids, saccharides, buffer salts etc.)
e.g. Chocolate agar (supplemented with blood and
slowly heated at 80 0 C in order to lyse erithrocytes –
chocolate colour); maybe prepared to be selective (by
adding antimicrobials) = Thayer Martin transparent
colonies
Neisseria
meningitidis
 Detailed biological and
medical definition
• Meningococci are bacteria that maybe
involved in medium or high severity
infections, recognizing respiratory mucous
surface as entry portal. May cause:
• meningitis, mostly in small children (6 months
- 1 year of age) and teenagers, immune
depressed patients with native or adaptive
immune response deficit etc.
• arthritis,
• Other infection sites at the mucous or serous
surfaces level
• pneumonia, septicemia
 Meningococcal disease
• Meningococcal meningitis – of variable severity,
pending on the immune state of the host
- 1/1000 carriers get ill
- fatality rate: 5-15% (WHO)
- skin damage (petechial or polimorphic purpuric
eruption) in 75% of cases
- Clinical forms:
- usually light or medium severity
- fulminant form: Waterhouse – Friedericksen
syndrome (endotoxic origin)
Petechial eruption in a
meningitis case
Other N. meningitidis infections
• Rare cases of conjunctivitis, synusitis,
pneumonia, urethritis

• Chronic manifestations - meningococcemia

with:
– Meningococcal arthritis
– Endocarditis
 IVa. Neisseria meningitidis:
– Gram negative cocci, tend to resist to
decolorisation with acetone-alcohol, non-
sporulated, non-mobile
Usually arranged in pairs, coffee beans shaped,
flattened or concave adjacent sides
– Oxidase and catalase positive; strictly
aerobic
– Carboxyphilic (capnophilic),
– Very fragile in exterior environment – need
constant 37 0 C temperature (mezophilic)
 Metabolic type and enzymatic
properties

 HETEROTROPHIC metabolic type (grow only in

the presence of organic material)
 Fastidious
 Oxydase and catalase positive; strictly aerobic
bacteria
 Relatively reduced saccharolytic spectrum:
Glucose, Maltose
(ǂ gonococcus/only Glucose)
Resistance to physical, chemical
and biological agents
• Physical agents:
• Very sensitive to:
– cold (Neisseria die quickly at temperatures under 37 0
C)
– dryness
• Consequences:
– Contamination is only direct: dangerous zone – 2-3 m
around the patient (Pflugge drops)
– Need quick transport in a 37 0 C warmed container (Mx
1 hour) or direct inoculation
– You must never refrigerate samples until inoculation !!!
 Pathogenicity factors
• Fimbria – adherence to mucosal epithelia and serous
surfaces
• Capsule:
- inhibits phagocytosis
- epidemiology: 12 groups (B,C,A,Y 95%)
• Outer Membrane Proteins (OMP) – types (P class 2,3),
subtypes (P1)
• LPS Antigenes: lipid A and oligosaccharides core
(external membrane, hyperproduced during
multiplication)
- intravascular disseminated coagulation syndrome
(IVDC Waterhouse – Friedericksen)
- epidemiology: immunotypes
• IgA – proteases – distroy local defence factors
 Pathogenesis:
• Colonisation of superior respiratory tractus
(adhesion pili)
• Reaches bloodstream (septicemia,
endotoxemia)
• Meningeal tissue and vascular damage
• Vasculitis – skin eruption
 Immune response: group
specific

• There is the possibility to repeat the

disease with another strain of
Meningococcus, belonging to another
antigenic group!!!
 Elements of epidemiology
- Humans only natural hosts
- Person-to-person transmission by aerosolization of
respiratory tract secretions in crowded conditions
- Close contact with infectious person (e.g., family
members, day care centers, military barracks, prisons,
and other institutional settings)
- Population groups with high receptivity: children of 6
month -1 year of age, immunosuppresed patients, half-
closed colectivities
- Endemic zone – meningitis belt – Sub Saharan Africa
- Romania:
- Sporadic or epidemic cases; cicles of 10-11 years
- Epidemic serogroups: A and C; group B mostly
sporadic
Sub Saharan Africa
meningitis belt
 Prophylaxis
• Indication of immunisation in Romania:
• Entire population for stopping group A and C
epidemics
• Travelling in hyperendemic countries
• Contacts

• Existing vaccine types:

– bivalent: A, C
– tetravalent: A, C, Y, W135
Neisseria
gonorrhoeae
 Detailed biological and
medical definition
• Gonococci are Gram negative cocci
involved in urogenital tract infections
• Gonococcal ophthalmia - rare
• Untreated, gonococcal infections evoluate
towards fibrotic chronicisation: local
fibrosis (e.g. arthritis, perihepatitis etc.)
Acute gonococcal diseases
– Gonorrhea
 Urethritis in men (with possible extension to
epididymis and prostate)
 Vaginitis in women: most often asymptomatic, with
possible extension to uterus, mostly asymptomatic,
extension towards uterus and fallopian tubes -
infertility or ectopic pregnancy, untreated develop
Pelvic Inflammatory Disease
 Conjunctivitis of newborns, opthalmia neonatorum
– rare cases of pharyngitis, sinusitis, pneumonia
– Very rare – generalized infection, resembling
meningococcal infection
Habitat of Neisseria gonorrhoeae
Human is only the natural host.

Found in human urogenital tract.

Rectal carriage in healthy individuals. !! MSM

Typically seen in pus cells (intracellular)

Found intracellularly in polymorphonuclear

leukocytes, or a specific category of white blood
cells with varying shapes of nuclei.
 Neisseria gonorrhoeae
– Morphological characters,
tinctoriality and cultivability resemble
those of N. meningitidis
– Resistance to physical, chemical,
biological agents – the same with
N. meningitidis except
N. gonorrhoeae is more often resistant to
Penicillin than N. meningitidis
 Pathogenic factors

• Fimbria, proteic adhesin – ensure adherence

to epithelia, especially uro-genital, but also other
mucous and serous surfaces
• Capsule:
– clinical: inhibits phagocytosis
• LPS antigen – only exceptionally involved in
IVDC syndrome (Waterhouse – Friedericksen)
• Iron chelator – ensure multiplication in local
sideropenia conditions
 Pathogenicity
• Colonization of uro genital tractus (adhesion pili,
proteic adhesine - adherence to mucosal epithelium)
- Cervix (the female) and urethra (the male)

• Invasion by contiguity (penetration into and

multiplication before passing through mucosal
epithelial cells), very rare septicemia, endotoxemia
and skin manifestation
 Immune response

• Usually local, not always efficient

• Possible chronicisation
 Epidemiology and prophylaxis
 Seriously underreported sexually-transmitted disease
 General receptivity
 Asymptomatic cases, especially in women – high risk of
transmission - major reservoir
 Infection transmitted, possibly together with other STD:
syphilis, Chlamydia trachomatis infection, HIV infection – need
for complex surveillance!
• Lack of protective immunity and therefore reinfection, partly
due to antigenic diversity of strains
Prophylaxis:
- comportamental education, use of condoms
- follow-up screening of sexual partners,
- antimicrobial prophylaxis for newborns ophtalmia
 Treatment
Antimicrobial treatment conducted by the specialist doctor
– Uncomplicated infection: ceftriaxone, cefixime or
fluoroquinolone

– Combined with doxycycline or azithromycin for dual

infections with Chlamydia

– Treatment of newborns with opthalmia neonatorum with

ceftriaxone
Penicillin is no longer drug of choice due to:
- Continuing rise in the MIC
- Plasmid-encoded beta-lactamase production
- Chromosomally-mediated resistance
 Laboratory diagnosis
1. Sampling
– Type of samples:
Meningococcus: CSF, blood, nose and throat
exsudate, skin lesions biopsy, articular fluid,
conjunctival secretion, sputum etc.
Gonococcus: endocervical secretion, urethral
secretion, anal swab, pharyngeal swab, conjunctival
swab, blood, articular fluid;
Genital secretion may be stimulated by mechanical,
thermal or biological (vaccin) means;
For women better results by the end of mensual
blood loss
Precautions: before starting antimicrobial therapy
 Transport
• Rapid transport, at 370C
• For samples other than blood - variants:
- direct inoculation in biphasic medium or
on plates with suitable enriched medium and
transport in CO2 enriched atmosphere
- Amies or Stuart transport medium
Disadvantage: direct examination is no longer
possible
• Do not refrigerate!
WHO Trans-isolate Medium
 Direct examination

• Bacterioscopic
– Stains : Gram,
methilen blue, M-G-G
– Quantitative (count)
and qualitative
cytology
• Rapid tests : latex or
co agglutination,
Immune-
electrophoresis
 Isolation
• Special media, 24-48 h, 37 0C, moist
atmosphere containing 5-10% CO2
- selective with antimicrobials
supplements (contaminated products)
- non neselective (blood, CSF etc.)
 Identification 
tion
– Clinical scope (case confirmation)
• Microscopy
• Growth appearance
• Oxidase positive
• Biochemical identification
(including rapid tests and
chromogenic media)
Neisseria meningitidis Neisseria gonorrheae
on chocolate agar on Thayer Martin agar
Positive oxidase reaction in
Neisseria

Kovac’s oxidase test: a negative and positive reaction on filter paper.

 Saccharolytic spectrum –
Neisseria meningitidis
Glucose and Maltose positive

Acid from oxidation of carbohydrates, not from

fermentation
 Identification 
tion
Epidemiologic scope
– Serotyping of N. meningitidis: groups,
types and subtypes
– Molecular typing
 Other tests
• Antimicrobial resistance
Penicillin resistance:
- rare in N. meningitidis
- frequent in N. gonorrhoeae
• Serological diagnostic (searching for
specific Ab by CFR) in chronic
gonococcal infections
 Key words N. meningitidis:

• Meningitis
• Waterhouse-Friedericksen syndrome
• Gram negative coccus – fastidious, coffee
bean shaped
• Capnophilia
• Thermophilia
• Oxidase positive
• Saccharolytic spectrum: Glucose, Maltose
• Chocolate Agar without antimicrobials for
CSF
• Capsule – A, C serogroups
• Vaccination
 Key words N. gonorrhoeae:
• Genital infections
• Gram negative cocci – fastidious, coffee bean
shaped
• Capnophilia
• Thermophilia
• Oxidase positive
• Saccharolytic spectrum: Glucose
• Chocolate Agar with antimicrobials (Thayer-Martin
chocolat agar)
• Complex investigation for detecting co-existence of
other STD: syphilis, Chlamydia or HIV co-infections
• Antimicrobial Susceptibility Testing
• Serological diagnostic for chronic gonorrhea cases