Theme: Gramnegative cocci (gonococci,

meningococci).
Neisseria gonorrhoeae
N.gonorrhoeaе with fimbriae

Virulence
factors:
microcapsule,
fimbriae,
endotoxin, Neutrophils with
IgA protease intracellular gonococci
 N.meningitidis
(incomplete phagocytosis)
• The organism
tends to occur
intracellularly in
the cytoplasm of
neutrophyls
which are
attracted to the
site of
inflammation in
the meninges
Gonococcal pharyngitis Dermatitis-arthritis
syndrome

Ophtalmia neonatorum
 Therapy
• Cephalosporins (cefrtiaxone, cefixime)
inhibit cell wall synthesis.
• Fluoroquinolones (ciprofloxacine) inhibit
DNA replication (DNA-gyrase).
Neisseria meningitidis
N. meningitidis on chocolate agar
N. meningitidis is spread through saliva and respiratory secretions during
coughing, sneezing, kissing, chewing on toys 
 Life cycle of N. meningitidis
• The human nasopharynx is the sole natural
reservoir of N. meningitidis.
• From there, bacteria colonize the mucosal surface
as a commensal and can disseminate to the next
host. They are sometimes found to cross the
mucosal barrier and reach the underlying blood
vessels, where meningococcal disease occurs.
N. meningitis is the main cause of bacterial meningitis in children and young
adults, causing developmental impairment and death in about 10% of cases
and, furthermore, 10-20% of survivors suffer from lifelong sequelae such as

deafness, learning disabilities or limb amputations. 

 Systemic infections
• Meningococcemia – bacteria in blood.

• Meningitis - inflammation of meninges.

• Neisseria meningitidis
proliferates and
eventually fills the lumen
of blood capillaries with
multicellular aggregates.
• Pili are central virulence
factors required
for bacterial
adhesion along
the endothelium and
subsequent formation of
intravascular
aggregates.
• This vascular
colonization process is a
prerequisite for vascular
damage observed
during infection
 Control
• Groups AC, and ACYW135 capsular
polysaccharide vaccines are available.
 Therapy of meningococcal infection
• Penicillin is the drug of choice.
• Although penicillin does not penetrate the normal
blood-brain barrier, it readily penetrates the blood-
brain barrier when the meninges are acutely
inflamed.
• Chloramphenicol or a third-generation
cephalosporin (cefotaxime or ceftriaxone) is
used in persons allergic to penicillins.
 Laboratory diagnosis of Neisseria
• Microscopy - intracellular gram negative diplococci
• IF, PCR.
• Culture:
– To isolate N. meningitidis, cultures are taken from the nasopharynx, blood,
CSF, and skin lesions.
– To diagnose genital gonorrhea in males, the sample is taken from the urethra,
in females - from the cervix and the rectum.
Identification of colonies by:
– oxidase test,
Oxidase test
– gram-staining,
– carbohydrate fermentation:
• N. gonorrhoeae ferments only glucose
• N. meningitidis ferments glucose and maltose
• Precipitation test, ELISA - detection of meningococcal
antigen on cerebrospinal fluid or from skin lesions for rapid
identification
• CFT - diagnosis of chronic gonorrhea.
 Swab from urogenital tract,
conjuctive, urine Serum from
patient

Microscopic Bacteriological Serological

method method method
• Laboratory
diagnosis of
Primary inoculation in
Gram staining
the serum agar and CFT gonorrhoea
other special media

RESULT RESULT

Cultural
Inoculation a part of the colony
properties
in slanted serum agar
(pure culture accumulate)
Preparation,
Gram staining
Inoculation of the
Hiss medium

Investigation of sensitivity to antibiotics

END RESULT
Positive Direct Fluorescent Antibody Test
for Neisseria gonorrhoeae for Neisseria meningitidis
 Cultivation
1. Non-selective media – chocolate agar, blood
agar, serum agar, ascitic fluid agar.
• Appropriate when plating samples from sterile body
sites (blood).
2. Selective medium - Martin Thayer Medium (MTM)
• Contains antibiotics to inhibit normal body flora:
– vancomycin to inhibit gram-positive bacteria;
– colistin to inhibit gram-negative bacteria;
– trimethoprim to suppress Proteus;
– nystatin to inhibit yeast.
• Appropriate for plating samples from complicated
niches (endocervix, urethra).
Colonies of N.gonorrhoeae
 Colonies of N.meningitis
on MTM chocolate agar on blood agar
Identification of Neisseria: Carbohydrate Utilization
• N. meningitidis produces acid from oxidation of glucose and
maltose, but not from sucrose or lactose. (A)
• N. gonorrhoeae produces acid from oxidation of glucose but not
from maltose, sucrose, or lactose. (B)
• The acid turns the pH indicator phenol red from red to yellow.

A B
CFT with gonococcal antigen
 Principle of CFT
• It is the nature of the complement to be activated when
there is formation of antigen-antibody complex.
 Methodology of
CFT
• The first step is to heat the serum at
56°C to destroy patient’s
complement.
• A measured amount of complement
and antigen are then added to the
serum.
• If there is presence of antibody in
the serum, the complement is fixed
due to the formation of Ag-Ab
complex.
• If no antibody is present then the
complement remains free.
• To determine whether the
complement has been fixed, sheep
RBCs and antibodies against sheep
RBCs are added (Hemolytic or
indicator system).
 Complement fixation test
In a positive
test
erythrocytes
During a
settle to the negative
bottom, and reaction the
the liquid becomes
supernatant pink (lacquered)
without any
liquid is precipitate of
transparent erythrocytes.
and colourless
• 1. Gonorrhea was revealed in the patient
on bacterioscopy of the smear from
urethra. Taking into account that
medecines for gonorrhea are
fluorquinolones, patient should be
prescribed:
• A. Ciprofloxacin.
• B. Fluorouracil.
• C. Cefazoline.
• D. Urosulfan.
• E. Furazolidone.
• 2. A doctor made the diagnosis of
gonorrhea. It was known from the
anamnesis that a patient had had
gonorrhea before and he had been treated
completely. What type of infection can this
new disease be attributed to?
• A. Secondary infection.
• B. Superinfection.
• C. Autoinfection.
• D. Reinfection.
• E. Relapse.
• 3. The most common route of entry into
the central nervous system by bacteria to
cause meningitis is:
• A. Through the middle ear.
• B. Through the cribiform plate.
• C. From the blood.
• D. Through the optic nerve.
• E. Through cranial wounds.
• 4. Damage caused by Neisseria
gonorrhoeae is associated with:
• A. Pili.
• B. Opa protein.
• C. IgA protease.
• D. Lipopolysacharide.
• E. Polysaccharide capsules.
• 5. A Gram-negative bean-shaped diplococcus is
isolated from a blood culture bottle. It grows on
pepton-blood base medium or Thayer-Martin
agar, produces acid from glucose and maltose,
and cannot grow on the meat pepton agar. The
most likely identity of this isolate is:
• A. Neisseria gonorrhoeae.
• B. Neisseria meningitidis A.
• C. Staphylococcus epidermidis.
• D. Streptococcus pneumoniae.
• E. Streptococcus pyogenes.
• 6. The cause of the meningococcal
meningitis in those susceptible spread
from………………results in……………,
which is usually followed by rapid
involvement of the meninges.
• A. Gut, septicaemia.
• B. Skin, pyaemia.
• C. Lungs, pneumonia.
• D. Food, food poisoning.
• E. Nasopharinx, septicaemia.
• 7. Gonococci and meningococci can be
differentiated from each other by:
• A. Gram’s stain.
• B. Oxidase test.
• C. Growth in CO2.
• D. Sugar fermentation.
• E. Capsule production.
• 8. Laboratory diagnosis of gonorrhea is
based on, EXCEPT:
• A. Immunofluorescence.
• B. Isolation of bacteria on serum agar.
• C. Gram’s stain.
• D. CFT.
• E. Isolation in mice.
• 9. Two children from kindergarten were
delivered to a hospital with diagnosis
“meningococcal meningitis”. It is necessary to
examine other children and staff for
meningococcal carriage. What method of
diagnosis should be used?
• A. Microscopy of CSF.
• B. Detection of meningococcal antigen in CSF.
• C. Cultural research of nasopharyngeal swab.
• D. Detection of titer of antibodies.
• E. Skin allergic test.
• 10. Gram-negative diplococci were isolated from
the nasopharynx of a boy with suspected
diagnosis “meningitis”. Which of the following
facts suggest that the isolated microorganism is
N. meningitides?
• A. It grows at 37 0C on serum agar.
• B. It produces pigment.
• C. It ferments sucrose.
• D. It grows on MPA.
• E. It does not grow at 37 0C on blood agar.