Pathogenic Gram negative cocci advanced bact.ppt

bacteriology

1. CHAPTER IV
PATHOGENIC GRAM NEGATIVE
COCCI
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2. Learning objectives
At the end of this chapter, the student will be able to:
1.Discuss the general characteristics of the genus Neisseria.
2.List the virulence factors of N. gonorrhoea.
3.Discuss the pathogenesis and clinical manifestation of N.
meningitidis.
4. Differentiate N. gonorrhoea from N. meningitidis using
biochemical tests..
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3. Genus Neisseria
General characteristics
They are non-motile, gram-negative intracellular
diplococci
Rapidly killed by drying, sunlight, heat, and disinfectants
Ferment carbohydrate and produce acid but not gas
Are kidney-shaped with adjacent concave sides.
Grow best on complex media under aerobic conditions
containing:
Heated blood, hemin, animal protein, 3- 10% Co
2 and other
supplements.
Are oxidase positive.
The main species of medical importance are:
- N. meningitidis
- N.gonorrhoeae.
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4. 4.1 Neisseria gonorrhoeae
The name ‘gonorrhoea’ derives from the Greek words
gonos ( seed) and rhoia ( flow), and described as a
condition in which
Semen fluid from the male organ without erection.
General characteristics
An obligate parasite of the human urogenital tract.
Has no polysaccharide capsule but has multiple serotypes
based on the antigencity of its pillus 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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5. 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)
 are also antigenic
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6. 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. Transferrin and lactoferrin binding proteins
Mediate acquisition of iron for bacterial metabolism.
5. Beta- lactamase
Hydrolyzes beta -lactam ring in penicillin.
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7. 6. Lipooligosaccharide (LOS)
Gonococcal lipooligosaccharides (LOS) have shorter,
highly branched, nonrepeat O-antigenic side chains
than do lipopolysaccharides found in other gram-
negative bacteria.
Is responsible for most of the symptoms / toxicity of
gonorrhea due to endotoxin 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.
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8. Pathogenesis and clinical manifestations
Gonococci attack mucous membrane of genito
urinary tract, eye, rectum and throat.
 Gonorrhea in adults is almost invariably transmitted by
sexual Intercourse and infants during birth process
1.Genitourinary tract infections:
– Symptoms are more acute and easier to diagnose in males.
–The patient typically presents with a yellow, purulent urethral
discharge and painful urination.
• In females,
– infection occurs in the endocervix and extends to the urethra
and vagina.
–
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9. •A greenish-yellow cervical discharge is most
common, often accompanied by intermenstrual
bleeding.
–The disease may progress to the uterus, causing
salpingitis (inflammation of the fallopian tubes), pelvic
inflammatory disease (PID), and fibrosis.
–Infertility occurs in approximately 20% of women with
gonococcal salpingitis, as a result of tubal scarring.
•Infant:
• (When delivered through the infected birth canal)-
Gonococcal ophthalmia neonatorum
–If untreated and complicated leads to blindness.
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10. •Rectal infections
–prevalent in male homosexuals,
–are characterized by constipation, painful
defecation, and purulent discharge.
•Pharyngitis Pharyngitis
–is contracted by oral-genital contact.
–Infected individuals may show a purulent
exudates, and the condition may mimic a mild
viral or a streptococcal sore throat.
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11. Laboratory diagnosis
Specimen: Urethral swab, cervical swab, eye swab.
Smear: Gram-negative intracellular and/or extra
diplococci.
Figure . ….. Left: N. gonorrhoeae Gram stain of pure culture
Right: N. gonorrhoeae Gram stain of a pustular exudates
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12. Culture:
Requires an enriched media like chocolate agar, Thayer-
Martin agar(TM) and/or Modified New York City
(MNYC)medium.
 Grows best in CO
2 enriched aerobic atmosphere with
optimal temperature of 35-37
0
c.
but some grow under anaerobic situations
On culture, Neisseriae species form convex, elevated
and mucoid colonies
On TM and Chocolate agar media, colonies are
transparent or opaque and non - pigmented.
•Ferment carbohydrate and produce acid without gas
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13. 13

14. Serology: Antibodies to gonococcal pili & outer
membrane proteins using RIA and/or ELISA.
Genetic probes : For detection of nucleic acids.
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.
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15. Prevention and control
•Avoid multiple sexual partner
•Using mechanical protection methods (condom)
•Early diagnosis and prompt treatment of cases
•Screening of high risk population groups.
Ophthalmic ointment application of
erythromycin or tetracycline to the conjunctiva
of all newborns
 
Health education
There is no effective vaccine to prevent
gonorrhea
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16. 4.2 Neisseria meningitidis
Characteristics
Gram-negative intra cellular diplococci.
has a prominent antiphagocytic polysaccharide capsule.
strains are grouped on the basis of their capsular
polysaccharides, into 12 serogroups,
some of which are subdivided according to the
presence of outer membrane protein and
lipopolysaccharide antigens.
•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.
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17. Epidemyology
The organism tends to colonize the posterior nasopharynx of
humans, and
humans are the only known host.
Present in the nasopharynx in 5-10% of healthy people
Transmission occurs through inhalation of respiratory
droplets from a carrier or a patient in the early stages of the
disease
Other risk factors include recent viral or mycoplasma upper
respiratory tract infection, active or passive smoking, and
complement deficiency.
In susceptible persons, pathogenic strains may invade the
bloodstream and cause systemic illness after an incubation
period of 2 to 10 days.
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18. Virulence factors
1. Meningoccal endotoxin (LOS) : is responsible for many toxic
effects.
2. Capsule : Protects bacteria from antibody mediated
phagocytosis.
3. Pili : Allow to colonization of nasopharynx.
Pathogenesis and clinical
manifestations
Infection with N. meningitidis has two presentations:
1.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 oropharyngeal
mucosa, cross the mucosal barrier, and enter the bloodstream.18

19. 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.
2. meningococcal meningitis
The onset of meningococcal meningitis may be abrupt or
insidious.
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
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20. Fulimenant meningococci (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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21. Laboratory diagnosis
Specimen: Cerebrospinal fluid, blood
Smear: Gram-negative intracellular diplococci
Culture: Transparent or grey, shiny, mucoid colonies in
chocolate agar after incubation at 35-37
O
c in a CO2
enriched atmosphere.
 Serology: Antibodies to meningo-coccal polysacharides
can be measured using:
latex agglutination or
hemagglutination tests.
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22. Biochemical reactions
Treatment
Drug of choice: Penicillin, Chloramphenicol, Cefotaxime
ceftriaxone
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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23. Prevention and control
•Chemoprophylaxis and immunization are both used to
prevent meningococcal disease
–Either rifampin or ciprofloxacin can be used for
prophylaxis in people who have had close contact
with the index case
•Vaccination with polyvalent conjugate vaccine to high
risk groups.
• Avoidance of over crowding.
NB: Meningococcal meningitis occurs as epidemics in
Africa and named as Meningitis belt.
–N. meningitidis serogroup A is the cause of African
meningitis epidemic.
–During epidemics, the carrier state rises from 5-10%
to 70-80%.

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24. Quiz I
1.List at least 5 characteristics of Neisseria
2. List mode of transmission of both N. gonorrhoeae
and N.meningitidis
3.Mention approprate samples for diagnosis of N.
gonorrhoeae and N.meningitidis
4.Differentiate N. gonorrhoeae and N.meningitidis
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