Lecture 5

Cocci
 Content
• Staphylococci (pyogenic cocci and coagulase-negative staphylococcus)
• Morphology, culture, and biological characteristics of Staphylococcus aureus
• The virulence factors of Staphylococcus aureus and their effects ( including SPA, coagulase,
hemolysin, and enterotoxin)
• The diagnostic laboratory tests for Staphylococcus aureus and the principles of controlling
Staphylococcus infections
• Streptococcus (classification)
• Morphology, culture, and biological characteristics of Streptococcus
• The virulence factors of group A streptococcus, the pyogenic and non- pyogenic infections caused
by group A streptococcus, diseases caused by group B, D streptococcus and enterococcus
(streptolysin, pyrogenic exotoxin, invasive enzymes)
• The diagnostic laboratory tests for streptococcus and pneumococcus; antistreptolysin O test (ASO
test)
• Neisseria
• Classification of Neisseria (Neisseria meningitides and Neisseria gonorrhoeae)
• The biological characteristics and pathogenicity of - and immune response to - Neisseria
meningitides
• Principles of diagnostic laboratory tests, and principles of prevention and treatment of the diseases
caused by Neisseria meningitides
• Neisseria gonorrhoeae and infection
 Staphylococcus
• Sir Alexander Ogston, a Scottish
surgeon, first showed in 1880
that a number of human
pyogenic diseases were
associated with a
cluster-forming
micro-organism. He introduced
the name staphylococcus
(Greek: staphyle = bunch of
grapes; kokkos = grain or berry), Alexander Ogston
now used as the genus name
for a group of facultatively
anaerobic, catalase-positive,
Gram- positive cocci.
 Classification
• Family: Micrococcaceae
• Genus:
o Staphylococcus
o Micrococcus
• Species:
S. aureus
S. saprophyticus
S. epidermidis
More: Micrococcus luteus than 20 species
 Genus Staphylococcus
• The genus Staphylococcus contains about forty
species and subspecies today.
• Only some of them are important as human
pathogens:
• Staphylococcus aureus
• Staphylococcus epidermidis
• Staphylococcus hominis
• Staphylococcus haemolyticus
• Staphylococcus saprophyticus
• other
 Structure and physiology
• Gram-positive cocci, nonmotile, facultative
anaerobes.
• Cells occur in grapelike clusters because cells
division occurs along different planes and the
daughter cells remain attached to one another.
• Salt-tolerant: allows them to tolerate the salt
present on human skin.
• Tolerant of desiccation: allows survival on
environmental surfaces (formites).
Staphylococcus aureus - Morphology

• Polysaccharide capsule is
only rarely found on cells.
• The peptidoglycan layer
is the major structural
component of the cell
wall. It is important in the
pathogenesis of
staphylococcal infections.
Other important
component of cell wall is
teichoic acid.
 Protein A
• Protein A is the major protein component of the cell wall. It is located on the cell
surface but is also released into the culture medium during the cell growth.

• A unique property of protein A is its ability to bind to the Fc part of all IgG
molecules except IgG3. It is not an antigen-antibody specific reaction.

• The surface of most S. aureus strains (but not the coagulase-negative staphylococci)
is uniformly coated with protein A.

• The presence of protein A has been exploited in some serological tests, in which
protein A-coated S. aureus is used as a nonspecific carrier of antibodies directed
against other antigens.

• Additionally, detection of protein A can be used as a specific identification test for S.

aureus.
Protein A binding
 Peptidoglycan
• Half of the cell wall by weight is peptidoglycan, a
feature common to gram-positive bacteria.
• The subunits of peptidoglycan are
N-acetylmuramic acid and N-acetylglucosoamine.
• Unlike gram-negative bacteria, the peptidoglycan
layer in gram-positive bacteria consists of many
cross-linked layers, which makes the cell wall
more rigid.
 Teichoic acid
• Teichoic acid is species-specific, phosphate-
containing polymers that are bound covalently
to the peptidoglycan layer or through
lipophilic linkage to the cytoplasmic
membrane (lipoteichoic acid - LPA).
• Teichoic acid mediates the attachment of
staphylococci to mucosal surfaces through its
specific binding to fibronectin.
 Coagulase and other surface proteins

• The outer surface of most strains of S. aureus

contains clumping factor (also called bound
coagulase).
• This protein binds fibrinogen, converts it to
insoluble fibrin, causing the staphylococci to
clump or aggregate.
• Detection of this protein is the primary test for
identifying S. aureus
 Coagulase binding test
Coagulase - cause blood to coagulate
Blood clots protect bacteria from phagocytosis from WBC’s and
other host defenses .
Staphylococci - are often coagulase positive
Negative
The genus Staphylococcus can be
divided into two subgroups (on the
basis of its ability to clot blood
plasma by enzyme coagulase):
coagulase-positive: Staphylococcus
aureus
coagulase-negative:
oStaphylococcus epidermidis,
o Staphylococcus hominis,
o Staphylococcus haemolyticus,
o Staphylococcus saprophyticus,
o Staphylococcus simulans,
o Staphylococcus warneri
o other
Positive
 Other surface proteins
• Other surface proteins that appear to be
important for adherence to host tissues
include:
collagen-binding protein
elastin-binding protein
fibronectin-binding protein
 Capsule
• Capsule or polysaccharide slime layer
• A loose-fitting, polysaccharide layer (slime layer) is only
occasionally found in staphylococci cultured in vitro, but is
believed to be more commonly present in vivo.
• Eleven capsular serotypes have been identified in S.
aureus, with serotypes 5 and 7 associated with majority
of infections.
 Capsule
• Capsule helps Staphylococcus.
• The capsule protects the bacteria by inhibiting
the chemotaxis and phagocytosis of
staphylococci by polymorph nuclear
leukocytes, as well as by inhibiting the
proliferation of mononuclear cells.
• It is also facilitates the adherence of bacteria
to catheters and other synthetic materials.
 Cytoplasmic membrane
• The cytoplasmic membrane is made up of a
complex of proteins, lipids, and small amount
of carbohydrates.
• It serve as an osmotic barrier for the cell and
provides an anchorage for the cellular
biosynthetic and respiratory enzymes.
 Enzymes
• Staphylococcal enzymes:
Coagulase
Catalase
Hyaluronidase
Fibrinolysin
Lipases
Nuclease
Penicillinase
 Role of Enzymes
1. Coagulase – Triggers blood clotting
2. Hyaluronidase – Breaks down hyaluronic acid,
enabling the bacteria to spread between cells
3. Staphylokinase (Fibrinolysin) – Dissolves fibrin
threads in blood clots, allowing Staphylococcus
aureus to free itself from clots
4. Lipases – Digest lipids, allowing staphylococcus to
grow on the skin’s surface and in cutaneous oil glands
5. β-lactamase – Breaks down penicillin – Allows the
bacteria to survive treatment with β-lactam
antimicrobial drugs
 Staphylococcal toxins
• S. aureus produces many virulence factors, including
at least five cytolytic or membrane-damaging toxins:
• alpha toxin =alpha hemolysin-pore forming toxin
lyses membrane, it is inactive against neutrophils
• beta toxin
• delta toxin Cytotoxic
• gamma toxin factors
• Panton-Valentine toxin =kleucocidin –
pore forming toxin – causes tissue necrosis,
active against neutrophils.

• two exfoliative toxins

• eigth enterotoxins (A-E, G-I)
• Toxic Shock Syndrome Toxin 1 (TSST-1)
Toxic Shock Syndrome Toxin and other
Toxins
• The enterotoxins and TSST-1 belong to a class
of polypeptide known as super antigens (SAgs)
• Non-specific binding of toxin to receptors
triggers excessive immune response
• Staphylococcus aureus strains produce several
other extracellular, biologically active
substances, including proteases,
phosphatases, lipases, lysozyme etc.
 Exfoliative toxins
• Exfoliatin splits interdermal
juntions
• Exfoliative toxins A and B results
in staphylococcal scalded skin
syndrome; usually in infants and
neonates.
• Staphylococcal scalded skin
syndrome (SSSS), a spectrum of
diseases characterized by
exfoliative dermatitis, is mediated
by exfoliative toxins.
• The prevalence of toxin
production in S. aureus strains
varies geographically but is
generally less than 5% to 10%.
 Food poisoning Enterotoxins
• Eigth serologically distinct staphylococcal
enterotoxins (A-E, G-I) and three subtypes of
enterotoxin C have been identified.
• The enterotoxins are stable to heating at 100 C
for 30 minutes and are resistant to hydrolysis
by gastric and jejunal enzymes.
• In preformed food heat-resistant enterotoxin
mediates staphylococcal food poisoning
(symptoms in 2-6 hours; usually self-limiting).
• Symptoms: nausea, abdominal cramping,
vomiting, and diarrhea.
• Foods particularly effective carriers of
bacteria and its toxins:
Custard of cream filled bakery food
Processed meat, chicken
Salads with mayonnes
Ice-cream, milk, dairy products
 Food poisoning Enterotoxins
• Thus, once a food product has been
contaminated with
enterotoxin-producing staphylococci
and the toxin have been produced,
neither reheating the food nor the
digestive process will be protective.
• These toxins are produced by 30% to
50% of all S. aureus strains.
• Enterotoxin A is most commonly
associated with disease.
• Enterotoxins C and D are found in
contaminated milk products, and
enterotoxin B causes staphylococcal
pseudomembranous enterocolitis.
 Toxin-mediated infections
• Toxic shock syndrome toxin (TSST-1) is
a super-antigen (SAg) capable of
activating large number of T cells.
• Was associated with use of tampons
but is also known to be associated
with postoperative wound or soft
tissue infections.
• The disease is characterized by high
fever, vomiting, diarrhea, sour throat
and muscle pain. Within 48 h it may
progress to severe shock with
evidence of renal and hepatic
damage. A skin rash may develop,
followed by desquamation at a
deeper level than in scalded skin
syndrome. Blood cultures are usually
negative.
 Toxic Shock Syndrome Toxin - 1
• TSST-1, formerly called pyrogenic exotoxin C and
enterotoxin F, is a heat and proteolysis resistant,
chromosomally mediated exotoxin.
• The ability of TSST-1 to penetrate mucosal
barriers, even though the infection remains
localized in the vagina or at the site of a wound, is
responsible for the systemic effects of TSS.
• Death in patients (5%-10%) with TSS is due to
hypovolemic shock leading to multiorgan failure.
 Epidemiology of Staphylococcus aureus
• Staphylococci are ubiquitous. All persons have
coagulase- negative staphylococci on their skin, and
transient colonization of moist skin folds with S. aureus
is common.
• Colonization of the umbilical stump, skin and perineal
area of neonates with S. aureus is common.
• S. aureus and coagulase-negative staphylococci are
also found in the oropharynx, gastrointestinal and
urogenital tract.
• Because staphylococci are found on the skin and in the
nasopharynx, shedding of the bacteria is common and
is responsible for many hospital-acquired infections
 Epidemiology

• Colonizes anterior nares of

20-30% of healthy people
• Strains with increased
virulence can not be
distinguished
• Community infections are
endogenoius
• Hospital spread is on the hands
of medical personnel
• Outbreaks involve nasal
carriers or workers with leason
• S. aureus survives drying
 Clinical Manifestations
• The clinical manifestations of some
staphylococcal diseases are almost exclusively
the result of toxin activity (e.g. staphylococcal
food poisoning and TSS), whereas other
diseases result from the proliferation of the
staphylococci, leading to abscess formation
and tissue destruction (e.g. cutaneous
infection, endocarditis, pneumonia, empyema,
osteomyelitis, septic arthritis).
 Staphylococcal Diseases
• Systemic Disease
• Toxic shock syndrome-TSS toxin is absorbed into the
blood and causes shock
• Bacteremia-presence of bacteria in the blood
• Endocarditis-occurs when bacteria attack the lining of
the heart
• Pneumonia-inflammation of the lungs in which the
alveoli and bronchioles become filled with fluid
• Osteomyelitis-inflammation of the bone marrow and
the surrounding bone
 Clinical diseases
Staphylococcal pyogenic infections:
• folliculitis,
• furuncle,
• carbuncle,
• bullous impetigo,
• panaritia and paronychia,
• wound infections,
• mastitis,
• osteomyelitis,
• staphylococcal pneumonia and other
 Staphylococcus infections

Bullos impetigo - Folliculitis - superficial

bubble-like swellings inflammation of hair follicle;
that can break and usually resolved with no
peel away; most complications but can
common in newborns
progress

Furuncullosis- boil; Carbuncles larger and deeper

inflammation of hair lesion created by aggregation
follicle or sebaceous gland and interconnection of a cluster
Panaritia Stye progresses into abscess or of furuncles
pustule
 Treatment
Antistaphylococcal antibiotics of the first choice:
• oxacillin (methicillin)
• cephalosporins of I. generation (cefazolin, cephalotin)
Antistaphylococcal antibiotics of the second choice:
• lincosamides (e.g. clindamycin)
• glycopeptides (vancomycin, teicoplanin)
• linezolid
• tigecyklin
• daptomycin
• and others
 Diagnosis
• Microscopy – smears of clinical materials are
stained according to Gram stain
• Cultivation on solid media (agar, usually blood
agar)
• Biochemical tests
• Phage typing – susceptibility of S. aureus
strains to various temperature phages
 Culture of Staphylococcus aureus
• Colonies on solid media
are round, regular,
smooth, slightly convex
and 2 to 3 mm in
diameter after 24h
incubation.
• Most strains show a
α-haemolysis surrounding
the colonies on blood
agar.
• S. aureus cells produce
cream, yellow or orange
pigment.
Laboratory identification scheme for Staphylococcus
Laboratory identification scheme for Staphylococcus
 Growth of Staphylococcus
• Staphylococcus species are facultative
anaerobic bacteria.
• All species grow best on nutrient agar and
blood agar.
• Mannitol salt agar is a selective media for
Staphylococcus species.
• It can grow at a temperature range of 15-45 C,
and NaCl concentration as high as 15%.
 Mannitol Salt Agar Not
Staphylococcus

Staphylococcus aureus

S. epidermidis
 Novobiocin test

S. saprophyticus S. epidermidis
 Resistance of Staphylococcus sp.
• Like most of medical important
non-spore-forming bacteria, S. aureus is
rapidly killed by temperature above 60 OC
• S. aureus is susceptible to disinfectants and
antiseptics commonly used.
• S. aureus can survive and remain virulent long
periods of drying especially in an environment
with pus
Antibiotic sensitivity pattern
 MRSA
• Methicillin-resistant S. aureus
• Resistant to all penicillins, cephalosporins,
and imipenems
• Usually multiply-resistant
• Vancomycin resistance is very rare – so far
Hospital-acquired
• Community-acquired cases now (CA MRSA)
 Panton-Valentine Leukocidin
• Panton-Valentine Leukocidin (PVL) consists of
2 components S and F, together with γ
exotoxin lyses WBC resulting in massive
release of inflammatory mediators responsible
for necrosis and severe inflammation
• PVL is an important virulence factor in MRSA
infections
Coagulase-negative staphylococcal spp
(CoNS)
• S. epidermidis – most
frequently isolated
staphylococcal spp.
• Colonizes moist body areas
such as axilla, inguinal and
perianal areas, anterior nares
and toe webs
• Important cause of nosocomial
infection esp. S. epidermidis
• Usually causes nosocomial
infections in patients with
predisposing factors such as S. epidermidis
immunodeficiency/
immunocompromised or
presence of foreign bodies
 Staphylococcus epidermidis
• Skin commensal
• Has predilection for plastic
material
• Associated with infection of IV
lines, prosthetic heart valves,
shunts
• Causes urinary tract infection in
catheterized patients
• Has variable antibiotic sensitivity
pattern
• Treatment should be aided with
antibiotic susceptibility test
Infections caused by S. epidermidis
• Blood stream infection
• Endocarditis
• Cerebrospinal fluid (CSF) shunt infection
• Peritoneal dialysis catheter infection
• Urinary tract infections, especially with indwelling urinary
catheters resulting to urinary tract complications
• Prosthetic joints infections
• Infection of vascular grafts
• Infection among newborns
• Eye infection after an eye surgery
• Infection of pacemakers or implantable cardioverter-defibrillators
• Infection of breast implants
 Importance of S.saprophyticus
• Skin commensal
• S. saprophyticus frequently isolated in rectum and
genitourinary tract of young women
• Can be causative agent in UTI in young healthy
women
• 2nd most common urinary pathogen (other than
E. coli) in uncomplicated cystitis in young women
• Colony counts of ≥ 105 CFU/ml usually indicative
of significant Bacteriuria
• Usually sensitive to wide range of antibiotics
Prevention
• Hand antisepsis is the
most important
measure in preventing
nosocomial infections
• Also important is the
proper cleansing of
wounds and surgical
openings, aseptic use of
catheters or indwelling
needles, an appropriate
use of antiseptic.
Streptococcus
 Streptococcus sp.
• Gram positive cocci, occur
in pairs and in short
chains.
• None-motile, none- spore
formers, some strains are
capsulated.
• Strict anaerobes and
facultative anaerobes.
• Gram positive
• Catalase negative
Streptococcus species are classified according
to hemolytic activity.
 Streptococci

Trypticase soy agar culture plate containing 5% sheep’s blood growing group-D Streptococci (left
wedge), group-B Streptococci (middle wedge), and group-A Streptococci (right wedge) bacteria.
Rebecca Lancelfield’s classification
Lancelfield’s antigens are cell wall carbohydrates

Group A – rhamnose-N-acetylglucosamine
Group B – rhamnose-glucosamine polysaccharide
Group C – rhamnose-N-acetylglucosamine
Group D – glycerol teichoic acid containing
alanine & glucose
Group F – glucopyrasonyl-N- acetylgalactosamin

•Presence of Lancefield antigens defines the

pyogenic streptococci
•Only pyogenic streptococci are β-hemolytic
•Hemolysis is the practical guide to classification.
 Classification - Lancefield
• Lancefield realized that all species in each “group”
generally (and conveniently) shared clinically
significant properties such as type of hemolysis,
normal host, body system or tissue where
indigenous, etc.
For example:
• Group A - S. pyogenes: human upper respiratory
• Group B - S. agalactiae: human urogenital
• Group C - S. zooepidemicus: from animal products
• Group D - S. faecalis: bile-resistant, fecal origin
Group Common Hemolysis Lencefield Surface Capsule Virulence Diseas
term cell wall protein factors
Streptococci

pyogenic
S. pyogenus GrA Strep β A M protein Hyaluronic M protein, Strep
acid lipoteichoic throat,im
(GAS) acid, go,pyoge
StrepSAgs, infection
streptolysin toxic sho
O,streptokina rheumat
se fever,glo
lonephri
S.agalactae GrBStrep β,- B - Salic acid Capsule Neonata
sepsis,m
(GBS) (9) itis, pyog
infection

S.equi β C - - StrepSAg genes Pyogenic

infections

S.bovis -, α D - - - Pyogenic

Other β, α,- E-W - - - Pyogenic

species
Pneumococcus

S.pneumoniae Pneumococcus α - Chlorine Polysaccharai Capsule, Pneumon

binding de (90+) pneumolysinn meningitis
euraminidase media, py
protein
infections
Group Common term Hemolysis Lencefield cell Surface protein Capsule Virulence Disease
wall factors

Streptococci

Viridans and
non-hemolyti
c
S. sanguis α Low virulence,
endocarditis

S.salivarius α Low virulence,

endocarditis

S. mutans α Dental caries

Other species α,- Low virulence,

endocarditis

Enterococci

E. faecalis Enterococcus -,α D Urinary tract , pyogenic

infections

E. faecim Enterococcus -, α D Urinary tract , pyogenic

infections

Other species -, α D Urinary tract , pyogenic

infections
 Pneumococcus and viridans
•Pneumococcus have an antigenic polysaccharide capsule

•Viridans and non-hemolytioc species lack Lancefield antigens

or capsules

•Streptolysin O and S cause β hemolysis

•Aerobically only S streptolysin is active

•Streptolysin O is active only in anaerobic conditions

 VIRULENCE FACTORS
1. Hyaluronic acid capsule -
nonimmunogenic
2. M protein – hair-like
projections on the cell wall
major virulence factor
promotes adherence
antiphagocytic
anticomplement
type specific
3. F-protein – fibronectin
binding protein
4. Lipoteichoic acid (LPA) –play
a role in pathogenesis
Cell wall structure
 Virulence Factors of β-Hemolytic
S. pyogenes
Produces surface antigens:
• C-carbohydrates – protect against lysozyme
• Fimbriae – responsible for adherence
• M-protein – contributes to resistance to
phagocytosis
• Hyaluronic acid capsule – provokes no immune
response
• C5a protease hinders complement and neutrophil
response, degrades complement
 Extracellular Virulence Factors
• Streptolysin O – a
pore-forming toxin– damages
membranes
• StrepSAgs i.e. Erythrogenic
toxins: are produced by some
strains, cause:
rash of scarlet fever ,
pyrogenicity, acute rheumatic
fever (ARF)
lethal shock – Streptococcal
Toxic Shock Syndrome (STSS)
Clinical significance of Streptococcus pyogenes:

• 1-Acute pharyngitis and

tonsilitis.
• 2-Impetigo and deep skin
infection.
• 3- Acute Rheumatic fever.
Due to the bacteria M
protein.
Streptococcus tonsilitis
• 4-Acute nephritis.
Due to antibodies
interaction with glomerulus.
Streptococcal tonsilitis
 Poststreptococcal Sequelae
• Acute rheumatic fever
(ARF) follows respiratory
infection
• Rheumatic heart disease
is produced by recurrent
ARF
• Glomeronephritis follows
respiratory or skin disease
• Only “nephritogenic”
strains are involved.
Streptococcal erysipelas
Neisseria
 Neisseria
• The Gram negative
Diplococci:
Neisseriae meningitidis
Neisseriae gonorrhoeae
• Gram negative cocci
• Kidney-shaped, occur in
pairs, in clinical
specimens: present inside
the polymorphonuclear
cells.
 Clinical significance
• Neisseriae meningitidis
• 1-Meningitis and Meningococcemia.
• 2-Septicemia.
• Neisseriae gonorrhoeae
• 1-Urinary tract infection: (Most common):
• A-Gonococcal Urethritis.
• B-Pelvic inflammatory disease.
 Neisseria infections
• N. meningitidis causes meningitis and
meningococcemia. It is the leading cause of
death from infection in children.
• N. meningitidis has a prominent
polysaccharide capsule that enhances
virulence by its antiphagocytic action.
 Neisseria gonorrhoea vs. N. meningitidis

• N. gonorrhoeae has no polysaccharide

capsule, it has three outer membrane proteins
(proteins I, II and III) protein II plays a role in
attachment of the organism to cells and varies
antigenically as well.
• The endotoxin of N. meningitidis is a
lipopolysaccharide (LPS) but the endotoxin of
N. gonorrhoeae is a lipo -oligosaccharide
(LOS).
Characteristics Neisseria gonorrhoeae Neisseria meningitidis
N. gonorrhoeae is the agent of N. meningitidis is a major cause of
Agents
gonorrhoea. cerebrospinal meningitis.

N. gonorrhoeae form smooth, round,

N. meningitides would form smooth,
moist, uniform
round, moist, uniform
Colony Morphology grey/brown colonies with a greenish
large grey/brown colonies with a glistening
colour underneath on primary isolation
surface and entire edges.
medium.

N. gonorrhoea is kidney shaped with N. meningitidis is semicircular diplococcus

Morphology
apposing ends concave. with flat apposing ends.

N. gonorrhoeae form smooth, round,

N. meningitides would form smooth,
moist, uniform
round, moist, uniform
Colony Morphology grey/brown colonies with a greenish
large grey/brown colonies with a glistening
colour underneath on primary isolation
surface and entire edges.
medium.

N. gonorrhoeae grow less N. meningitidis grow well on blood agar

Growth on Blood Agar
well on blood agar than N. meningitidis. than N. gonorrhoeae

Capsule No Yes

Pathogens It is always considered a pathogen. It is not always considered as pathogens.

N. gonorrhoeae infections have a high N. meningitidis infections have a low

Prevalence and Mortality
prevalence and low mortality prevalence and high mortality.

Cause meningitis and other forms of

N. gonorrhoeae can also cause
meningococcal disease such as
Pathogenesis conjunctivitis, pharyngitis, proctitis or
meningococcemia, a life-threatening
urethritis, prostatitis, and orchitis.
sepsis.
 I. Neisseria meningitidis - Pathogenesis and
Epidemiology

• Humans are the only hosts for meningoccocci.

• The organisms are transmitted by airborne droplets,
they colonize the membranes of the nasopharynx and
become part of the transient flora of the upper
respiratory tract.
• A polysaccharide capsule that enables the organism to
resist phagocytosis by polymorphonuclear leukocytes
(PMNs).
• From the nasopharynx, the organism can enter the
bloodstream and spread to specific sites such as the
meningis or joints, or be disseminated throughout the
body.
Meningococci have 3 virulence factors:

• Endotoxin (LPS) which causes fever and shock.

• An immunoglobulin A protease helps the
bacteria attaches to the membrane of the
upper respiratory tract.
• A polysaccharide capsule
 Diagnostic Laboratory Tests
• Specimens: Specimens of blood are taken for
culture and specimens of spinal fluid are taken for
smear, culture and chemical determinations.
• Nasopharyngeal swab cultures are suitable for
carrier surveys.
• Smears: Gram-stained smears of the sediment of
centrifuged spinal fluid often show typical
neisseriae within polymorphonuclear leukocytes
or extracellularly
 Laboratory tests
• Cerebrospinal fluid (C.S.F.) specimens
are plated on chocolate agar and
incubated at 37° C in an atmosphere
of 5% CO2. Presumptive colonies are
identified by Gram-stain and oxidase
test.
• Serology: Antibodies to
meningococcal poly-saccharides can
be measured by latex agglutination or
hemagglutination tests.
• Fermentation test: N. meningitidis
ferment both glucose and maltose.
Growth of Neisseria meningitis

Blood agar Chocolate agar

II. Neisseria gonorrheae - Pathogenesis and
Epidemiology

• Gonococci like meningococci cause disease

only in humans. The organism is usually
transmitted sexually. Newborns can be
infected during birth.
• Gonorrheae is usually symptomatic in men but
often asymptomatic in women.
 Neisseria gonorrheae

• Genital tract infections are the most common

source of the organism.
• Pili constitute one of the most important
virulence factors, because they mediate
attachment to mucosal cell surfaces and are
antiphagocytic.
• The endotoxin of gonococci is weaker than
that of meningococci.
• Gonococci have no capsules.
 Neisseria gonorrheae
• Gonococci cause both localized infections
usually in the genital tract, and disseminated
infections with seeding of various organs.
• Gonococci reach these organs via the
bloodstream (gonococcal bacteremia).
• Gonorrhoeae in men is characterized by
urethritis accompanied by dysuria and a
purulent discharge.
 Neisseria gonorrheae
• In women infection is
located primarily in the
endocervix, causing a
purulent vaginal
discharge and
intermenstrual bleeding
(cervicitis).
• The most frequent
complication in women
is ascending infection of
the uterine tubes
(salpingitis) which can
result in sterility.
 Infections caused by Neisseria
gonorrheae
• Other infected sites include the
anorectal area, throat and eyes.
• Anorectal infections occur in
women and homosexual men.
• In the throat, pharyngitis
occurs. Ophthalmia neonatorum

• In newborn infants, purulent

conjunctivitis (ophthalmia
neonatorum) is the result of
gonococcal infection acquired
from the mother during
passage through the birth
canal.
Gonococcal pharyngitis
 Diagnostic Laboratory Tests

• Specimens: Pus and secretions are taken from

the urethra, cervix, rectum, conjunctiva, or
throat for culture and smear.
• Smears: Gram-stained smears of urethral or
endocervical exudate reveal many diplococci
within pus cells.
 Neisseria gonorrheae
• Culture: immediately
after collection, pus or
mucus is streaked on
enriched selective
medium (e.g. modified
Thayer-Martin medium)
and incubated in an
atmosphere containing
5% CO2 at 37° C.
• Fermentation test: N.
gonorrhea ferment
glucose only.
 Neisseria gonorrheae
• They grow best on Thayer-Martin Medium;
Chocolate agar supplemented by vancomycin
and nystatin.
• Both factor X and V (hemoglobin, NAD) are
required for Neissereiae species cultivation.
• A 5-10 % CO2 is required for primary
cultivation.