CHAPTER – 4
PATHOGENIC GRAM NEGATIVE COCCI
BY Adimasu B.(MSC)
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LEARNING OBJECTIVES
At the end of this chapter, the student will be able to:
Discuss the general characteristics of the genus Neisseria.
List the virulence factors of N. gonorrhoea.
Discuss the pathogenesis and clinical manifestation of N.
meningitidis.
Differentiate N. gonorrhoea from N. meningitidis using
biochemical tests..
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General characteristics
They are non-motile, gram-negative intracellular diplococci
Ferment carbohydrate producing acid but not gas
Are kidney-shaped with adjacent concave sides.
Grow best on complex media under aerobic conditions
containing 5%co2
Are oxidase positive.
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The main species of medical importance are:
- N. meningitidis
- N.gonorrhoea.
Neisseria species grow best on media containing: Heated
blood, hemin, animal protein, 3- 10% Co2 and other
supplements.
The organisms are sensitive to: drying, sunlight, moist
heat and disinfectants.
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General characteristics
An obligate parasite of the human urogenital tract.
Has no polysaccharide capsule but has multiple serotypes based
on the antigencity of its pilus protein.
There is a marked antigenic variation in the gonococcal pili as a
result of chromosomal rearrangement.
More than 100 serotypes are known.
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Virulence factors
1. Pili
Hair like appendages that mediate initial attachment to non - ciliated
human cells ( e.g. epithelium of vagina , fallopian tube and buccal
cavity.
Made from pilin proteins.
Resistance to phagocytosis (interferes with neutrophil killing)
2. Por protein (Protein I)
Pores on the surface of bacteria through which nutrients enter the cell.
Promotes intracellular survival by preventing phago-lysosome fusion in
neutrophils.
3. Opa (Protein II)
Important for firm attachment and invasion of bacteria to host cells.
4. RMP (protein III)
Outer membrane protein found in all strains of N. gonorrhoeae
It is associated with por in the formation of pores in the cell.
Protects other surface antigens (por proteins and lipo oligosaccaride)
from bacterial antibodies.
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5. Lipooligosaccharide (LOS)
Is responsible for most of the symptoms / toxicity of gonorrhea
due to endo-toxin effect of LOS.
Gonococcal LOS triggers an intense inflammatory response.
Subsequent activation of complement, attraction and feeding by
phagocytes and the lysis of the phagocytes themselves,
contributes to the purulent discharge.
6. Transferrin and lactoferrin binding proteins
Mediate acquisition of iron for bacterial metabolism.
7. Beta- lactamase
Hydrolyzes beta -lactam ring in penicillin.
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Pathogenesis and clinical manifestations
Gonorrhea in adults is almost invariably transmitted by sexual
Intercourse
Gonococci attack mucous membrane of genito urinary tract,
eye, rectum and throat.
In males there is urethritis, with yellow, creamy pus and
painful urination.
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In females , primary infection is in the endo-cervix
and extends to the cervix and vagina giving rise to
muco-purulent discharge.
Gonococcal ophthalmia neonatorum is infection of
the eye of the new born during passage via infected
birth canal.
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Laboratory diagnosis
Specimen: Urethral swab, cervical swab, eye swab.
Smear: Gram-negative intracellular and/or extra diplococci.
Culture:
Requires an enriched media like chocolate agar
Thayer-Martin agar and/or Modified New York City medium.
Grows best in carbon dioxide enriched aerobic atmosphere
with optimal temperature of 35-370c.
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Growth characteristics
Most Neisseriae grow best under aerobic conditions but
some grow under anaerobic situations
Ferment carbohydrate and produce acid without gas
On culture, Neisseriae species form convex, elevated and
mucoid colonies
On Thayer - Martin and Chocolate agar media, colonies are
transparent or opaque and non - pigmented.
Serology: Antibodies to gonococcal pili & outer membrane
proteins using RIA and/or ELISA.
Genetic probes : For detection of nucleic acids.
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Treatment
Drug of choice (CDC) : Ceftriaxone, doxycycline, ciprofloxacin,
or oflaxacin. Penicillin resistance due to beta- lactamase
enzyme
producing N. gonorrhoeae have been identified.
For ophthalmia neonatorum - 1% silver nitrate , 1% tetracycline
or 0.5% erythromycine eye ointments.
Prevention and control
Early detection and treatment of cases.
Using condom
Health education
There is no effective vaccine to prevent gonorrhea
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Medically important sero-groups in humans disease are
encapsulated strains belonging to A, B, C, Y and W135.
Sero -groups C and A are associated with epidemic
disease.
Meningitis is the inflammation of the meninges of the brain
or spinal cord.
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Virulence factors
Meningoccal endotoxin (LOS) : is responsible for many toxic
effects.
Capsule : Protects bacteria from anti body mediated
phagocytosis.
Pili : Allow to colonization of nasopharynx.
Pathogenesis and clinical manifestations
Infection with N. meningitidis has two presentations:
Meningococcemia, characterized by skin lesions, and acute
bacterial meningitis
Infection is by inhlation of the bacteria, which attach to epithelial
cells of the nasopharyngeal and oro pharyngeal mucosa, cross
the mucosal barrier, and enter the bloodstream.
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The onset of meningococcal meningitis may be abrupt or
insidious.
Fulimenant meningo cocci (with/without meningitis) characterized
by:
-Fever
- Petechiae (minute hemorrhagic spots in the skin) or purpura
(hemorrhages into the skin). Occurs from first to third day of
illness in 30 to 60% of patients with meningococcal disease.
Pulmonary insufficiency developed within a few hours and
many patients die within 24 hours of being hospitalized.
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Meningitis begins suddenly with intense headache,vomiting,
photophobia, stiff of the neck or spinal rigidity (meningeal
irritation), neurologic signs (coma or convulsions) in 1/3 of
patients.
The most severe form of meningococcemia is the life threatening
Water House Friderichsen syndrome, which is characterized
by high fever, shock, widespread purpura, disseminated
intravascular coagulation and adrenal insufficiency.
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Diseases caused by Neisseria meningitidis includes :
Pyogenic (purulent) meningitis
Meningococcal bacteremia (Water House Friderichsen
syndrome)
Meningococcal encephalitis
Pneumonia
Arthritis and endocardiatis
Urethritis.
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Laboratory diagnosis
Specimen: Cerebrospinal fluid, blood
Smear: Gram-negative,cocci
Culture: Transparent or grey, shiny, mucoid colonies in
chocolate agar after incubation at 35-37Oc in a CO2
enriched atmosphere. intracellular diplococci
Serology: Antibodies to meningo- coccal polysacharides
can be measured using: latex agglutination or
hemagglutination tests.
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BIOCHEMICAL REACTIONS
Species Glucose Lactose Maltose Sucrose
Neisseria Positive Negative Negative Negative
gonorrhoeae
Neisseria Positive Negative Positive Negative
meningitidis
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Treatment
Drug of choice: Penicillin, Chloramphenicol, Cefotaxime
ceftriaxone
Prevention
Vaccination is available for sero groups A, C, Y and W135.
No effective vaccine for sero group B as it is poorly
immunogenic in humans.
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Refernce
1. Richard A. Harvey, Pamella C. Champ, Microbiology,
Lippincott’s illustrated reviews, 2 nd ed.
2. Benson’s microbiological application, Laboratory
manual in general microbiology, 8th ed . 2001
3. Sherris, Medical microbiology, an introduction to
infectious disease. 4th ed. 2004.
4. Monica Cheesbrough, District LaboratoryPractice in
Tropical Countries Part II. 2nd ed. 2006
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THANKS A LOT !
ANY QUESTION ?
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