MICROBIOLOGY A

NEISSERIACEAE-CHLAMYDIACEAE-MYCOPLASMACEAE

NEISSERIACEAE

*lifted from mra trans

* N. gonorrheae and N. meningitides -strictly human pathogens
*All Traditional pathogens (+) growth on the following selective media: ML (Martin Lewis Medium), MTM (Modified
Thayer Martin Medium), NYC (New York City Medium)
*All Commensals (-) growth on ML, MTM, NYC except N. lactamica (+ growth), N. cinerea and M. catarrhalis
(occasionally growth)
*catalase test is important in differentiation of Staphylococcus and Streptococcus
*all are catalase test (+) but the best run is an oxidase test because all Neisseria are oxidase (+), that is their initial
identification
*sugars are not fermented by Neisseria, they utilize sugars by oxidative reaction rather than fermentation
<sabi ni doc must know: sugars utilized by each pathogens>
No spores
GENERAL Catalase (+)
CHARACTERISTICS Oxidase (+) (reagent: tetramethyleneparaphenylene
HCl/oxidase reagent
Gram (-) diplococci in pairs (has flattened adjacent Often seen intacellularly, inside phagocytes (but
side, coffee bean shape they do not multiply intracellularly, they multiply
outside)

Non-motile (most cocci are non-motile)

Lecture Date: September 2015

Lectured by: Doc Natividad
[ppt-recordings-oldtrans]
 Commensals of oropharynx, nasopharynx, and
anogenital mucosal membranes
Oxidase Test
Procedure:
(1) Drop oxidase reagent on the filter paper (at
first no color)
(2) Put a sample of inoculum over the drop and
there will be change in color. Dark purple is
(+)
Another variation:
-reagent added medium to chocolate agar plate
-inoculate organisms
-Black indicates (+) (this black color is not a
pigment but a clor formed because of the reaction
Another variation:
-inoculate suspected organism
-place a disk of oxidase N (Neisseria test)
-incubate
-blackening circle indicates (+) oxidase test
GROWTH
Fastidous
Greyish, a little opaque colonies
Best under enriched medium with 5-10% CO2
Lecture Date: September 2015
Lectured by: Doc Natividad
[ppt-recordings-oldtrans]
-must be placed in a candle jar for CO2
retention or a CO2 incubator (not for
anaerobes, candle jar only decreases
oxygen tension and increases CO2
tension)
Very sensitive to adverse environment: drying,
sunlight, moist heat, many disinfectants
Do not refrigerate specimens: sensitive to cold
temperature
-if cant process right away, best to put in
incubator or at very least room
temperature
Chocolate Agar- medium of choice for normally
sterile specimens (from blood, CSF)
Both chocolate agar and blood agar have blood,
but the difference is the temperature at which blood
is added
-chocolate agar- 80oC (high temperature
lyse RBC making its content available to
organisms
-blood- 45oC (to not lyse RBC)
Thayer-Martin Medium- selective media
-For non sterile specimen (ex.
Nasopharyngeal discharge)
-inhibits growth of normal flora so only
Neisseria will grow
SELECTIVE MEDIA
(1) Thayer-Martin (TM)
Chocolate agar + VCN antibiotics
Vancomycin (for gram (+) bacteria)
Colistin (for gram(-) bacteria)
Nystatin (for fungi, especially candida,
best for urethral or rectal specimen)
Do not use as medium of choice because
some strains of Neisseria may be
unhibited by this. (MOC is still chocolate
agar)
(2) Modified Thayer-Martin Medium (MTMM)
Chocolate agar+ VCN+ trimethoprim
(3) Martin Lewis
(4) New York City
 Both 3 and 4 are chocolate agar with
particular antibiotics
A. NEISSERIA
GONORRHEAE
MOT
Very sensitive (once excreted/released from the
body, they are killed) to adverse environment: drying,
sunlight, moist heat, many disinfectants, therefore
mode of transmission is through close contact and not
through toilet seats or fomites
Sexual contact, birth canal for newborns (not
transplacental)
VIRULENCE FACTOR
(1) Pili/fimbriae
Attachment
Antiphagocytic (not by the capsule that
protects it but by the pili)
Ag variation of pili- reason why there is lack
of immunity, so can be reinfected
Therefore Neisseria without pili are not
virulent (they do not produce disease
(2) Por protein
Inhibits phagosome-lysosome fusion in
PMNs
Interfere degranulation of PMNs
Associated with disseminated disease (kasi
magcirculate si infected PMN)
N. gonorrhoea is not intracellular parasite,
once it is inside the PMNs, it is killed except
pag may Por protein
(3)OPA protein (opacity)
Firm attachment to eukaryotic cells
(4)RMP protein (reduction-modifiable protein)
Protects other surface proteins from
bactericidal Abs
(5)Proteins that mediate acquisition of iron:
Lecture Date: September 2015
Lectured by: Doc Natividad
[ppt-recordings-oldtrans]
a) Transferrin-binding protein
b) Lactoferrin-binding protein
*a and b sequester iron so that iron would not be
available for the organism
c) Hgb-binding protein
(6)Lipooligosaccharides (LOS)
Both N. gonorrheae and N. meningitides has
no O Ag; they have lipid A and core
polysaccharide. Lipid A is the one
responsible for endotoxic activity (pag
endotoxin-lipid and non immunogenic,
exotoxin-protein and immunogenic)
Endotoxin- non immunogenic because it
exhibits Ag mimicry
Ag mimicry- resembles human
glycospingolipid so they escape recognition
(7)IgA1 protease
Cleaves IgAI; further explains why there is
no permanent immunity against gonorrhoea
(8)Beta lactamase
Plasmid coded therefore may or may not be
present, used to be really sensitive to
penicillin
2 bacteria noted to have this, S. aureus and
N. gonorrheae
CLINICAL
MANIFESTATION
(1) Genital gonorrhea
a. Males: most males are symptomatic(95%);
characterized by urethritis accompanied by
dysuria and a purulent discharge
b. Females: 80% are asymptomatic; cervicitis
causing a purulent discharge and inter-
menstrual bleeding. May ascend to infect the
vagina causing vaginitis, caused by N.
Gonorrhea and Chlamydia
Vaginitis-usually caused by Candida
and Trichomonas, sometimes E. coli
 Vaginosis-no inflammation, caused
by Garnerella
c. DDx:
i. Chlamydia trichomatis NGU
ii. Ureaplasmin
iii. E. coli
(2) Rectal infection in homosexuals
(3) Pharyngeal infection
(4) Meningitis and conjunctivitis in adults
(5) PID (pelvic inflammatory disease)
(6) Disseminated gonococcal infection (arthritis,
dermatitis)-Especially if you have MAC deficiency
(7) Opthalmia neonatorum purulent conjunctivitis in
the newborn (erythromyocin in all newborns eyes)
To prevent from happening (since usually
mother is asymptomatic) give prophylaxis
to all newborns
(8) Vulvo-vaginitis
Gonorrhoea not normally a vaginitis and
if it is its usually cervicitis
Young children suggestive sexual abuse
Second most common cause of septic arthritis, first is S.
aureus
DIAGNOSIS
Gram stain or urethral or endocervical smear
Gram (-) diplococcic IC (sometimes EC)
presumptive
Culture
From men- not necessary when gram stain
is (+)
From women- a must, there are normal
flora that resembles N. gonorrhoea:
Moraxella osleonsis, Acitenobacter
calcoacetous (small g(-) coccobacillus,
both are EC
Nucleic Acid Amplification Tests (NAATs)
Rapid direct detection in genito-urinary
specimens
Not for extragenital specimens
Serology
Lecture Date: September 2015
Lectured by: Doc Natividad
[ppt-recordings-oldtrans]
Not useful
Serum and genital fluid contain IgG and
IgA Abs against gonococcal pili, outer
membrane proteins and
LPS(lipopolysaccharides)
Some IgM of human sera is bactericidal
for gonococcal in vitro
TREATMENT
DOC: Ceftriaxone
Add Doxycycline- for concomitant
chlamydia infection since it is usually
associated to gonococcal infection
Reason for giving more than 1 drug: N.
gonorrhoea is resistant to Doxyxycline
(so you can not give Doxycycline alone),
Chlamydia is resistant to Ceftriaxone (so
you can not give Ceftriaxone alone)
Penicillin used to be drug of choice but because of beta
lactamase/penicillinase, di na pwede
PPNG (Penicillinase Producing N. gonorrhoea)-
resistant to penicillin
Chromosomally mediated resistance to Tetracycline-
inherently acquired unlike those that are plasmid-coded
B. NEISSERIA
MENINGITIDES
MOT
Transmission through respiratory droplets (usually
large droplets so you have to be close to each other,
unlike TB, droplets are small so you dont need
close contact---pag malaking droplets kasi madali
syang babagsak from air, pag maliit, matagal syang
nasa air)
Humans are the only natural hosts
Does not last long so requires close contact
Very contagious, fatal, progressive infection
Colonizes nasopharynx of healthy people (carriers
harbour the organism in the nasopharynx)
 2nd most common cause of community-acquired
meningitis in adults (1st is S. pneumoniae
There can be swift pregression from good health to
life threatening disease
Highest incidence in children <5 yo and young
adults, institutionalized people(crowding), people
with late complement deficiencies (MAC)---from
C5 to C9
Tandaan, our protection from Neisseria is
complement and therefore antibodies too
Late complement (C5-C9)-for formation
of MAC which is responsible for cytolysis
Complement-mediated lysis is usually
ineffective in most bacteria. It is only
effective in Neisseria.
VIRULENCE FACTOR
(1) Capsule
Most important, without this, they cannot
cause disease
13 serogroups- most important in human
are A, B, C, Y and W-135 (group A
and C are associated with epidemic
disease)
Capsular Ags (polysaccharide) are found
in the blood and CSF (body fluids
(2) LOS- Endotoxin (lipid A)
No O Ag
Most potent endotoxin that is why thee is
rapid proliferation of disease
(3) IgA protease
Destroys IgA
CLINICAL
MANIFESTATIONS
(1) Septicemia (meningococcemia)
a. Thrombosis of small bvs: skin rashes,
Petechia, ecchymosis (all due to endotoxin)
b. Arthritis
Lecture Date: September 2015
Lectured by: Doc Natividad
[ppt-recordings-oldtrans]
c. Hemorrhage of the adrenals (Waterhouse
Friederichsen syndrome)
d. DIC, shock( endotoxin is cause of septic
shock), and death
(2) Meningitis
a. Fever, general malaise, nuchal rigidity
b. + Brudzinski sign, suggesting meningeal
irritation
c. CSF findings
i. 200-20k PMNs (normal: 0-5 lyms)
ii. glucose low (<45 mg/dL)
iii. protein high (>50mg/dL)
(3) Milder, chronic septicemia
(4) Other syndromes (Pneumonia, arthritis,
urethritis)
DIAGNOSIS
(1) Gram stain smear
a. Specimen:
b. Blood, CSF, petechial skin lesion
c. Nasopharyngeal swab = carrier
(2) Culture: BAP and CAP a. Oxidase test b. CHO
utilization test using CTA medi
productionin glucose and maltose
(3) Capsular polysaccharide Ag detection = CSF
or blood
a. Latex agglutination- more frequently used
b. CIE (Counter immune electrophoresis)
c. Limited value: serogroup B relatively non-
immunogenic
TREATMENT
DOC: Penicillin G
-very sensitive to Penicillin, no need for
susceptibility testing
Allergic: Chlorampenicol, 3rd generation
cephalosphorins (cefotaxime, ceftriaxone
PREVENTION
Chemoprophylaxis: rifampin, minocycline,
ciprofloxacin
-Eradicate carrier state
-Oral penicillin does not eradicate carrier
state
Vaccine: serogroups A, C, Y and W-135
-Tetravalent conjugate vaccine: capsular Ag
conjugated with diphtheria toxoid (T- dependent
response) because it is a hapten

ON A SCALE OF 1 TO 10,
HOW WOULD YOU RATE YOUR PAIN?

Lecture Date: September 2015

Lectured by: Doc Natividad
[ppt-recordings-oldtrans]