CHLAMYDIAE

Learning Objective
• Classification
• Morphology
• Life cycle
• Antigenic property
• Laboratory diagnosis
• pathogenicity
 Introduction
Chlamydia
• obligate intracellular bacterial parasite of
humans, animals, birds
• Tropism for squamous epithelium cells &
macrophages of RT, GIT
• Filterable & fail to grow in cell free media so
mistaken to be virus
 Introduction
Differ from virus-
• Have both DNA & RNA
• Have cell wall (inner and outer cytoplasmic
membrane), LPS & ribosomes
• Replicate by binary fission
• Susceptible to antibiotics &
chemotherapeutic agents
 Classification
• Family : Chlamydiaceae
• Genus : Chlamydia and Chlamydophila
• Chlamydia
- Chlamydia trachomatis
• Chlamydophila
– Chlamydophila pneumoniae
– Chlamydophila psittaci
 Morphology & Growth cycle
Chlamydia occurs in 2
forms-
1. Elementary body (EB)
2. Reticulate body/ initial
body (RB)
Elementary body (EB) -
• Extracellular, infective
form, non replicative
• Spherical or pear shaped
 Morphology & Growth cycle
• 200-300nm diameter
• Rigid trilaminar cell wall (same as GNB cell
wall)
• EB infect phagocytes, epithelial cells after
attachment to specific cell receptors
• Dense nucleoid present
 Morphology & Growth cycle
Reticulate body (RB) -
• Intracellular, replicative
form,
• 500-1000nm in size
• Cell wall fragile &
pliable
 Reproductive cycle of Chlamydia

• Growth cycle-
 Morphology & Growth cycle
EB enters the host cell by phagocytosis
Enlarges to form reticulate body (500-1000nm)
Divide by binary fission to produce large no of
elimentary bodies
Newly formed infectious particle (EB) gets
liberated
Infects new cell and cycle repeated
 Antigenic properties
3 major antigen present-
1. Genus specific antigen
• Heat stable
• Common to all chlamydia
2. Species specific Ag
• Present in all strains of chlamydia species
• Help classify chlamydia into species-
trachomatis, psittaci, pneumoniae, pecorum
 Antigenic properties
3. Intra- species typing antigen(serotype
specific Ag)
• Ag classified it into many serotypes.
• 2 important biovars of C.trachomatis - TRIC
(Trachoma,Inclusion Conjunctivitis) & LGV
(Lymphogranuloma venereum)
• TRIC biovars has 12 serovars- A, B, Ba, C, D-K
• LGV biovars has 3 serovars- L1, L2, L3.
 Human diseases caused by Chlamydia
S. No. Species Serotype Disease

1. C.trachomatis A, B, Ba, C Endemic blinding trachoma

2. C.trachomatis D, E, F, G, H, I, J, K Inclusion conjunctivitis

(neonate, adult); Genital
chlamidiasis; Infant
pneumonia

3. C.trachomatis L1, L2, L3 Lymphogranuloma venereum

4. C.psittaci Many serotypes Psittacosis

5. C.pneumoniae TWAR strain Acute respiratory diseases

 Lab diagnosis
Specimen collection
• All aseptic precaution taken & procedures
done in labs with suitable containment (Class 2
biosafety cabinets)
• Ocular, urethral, cervical specimens are
collected by mucosal scrapings
• Pus or discharge from bubo for LGV
• Blood, sputum, resp secretions for pneumonia
also collected
 Lab diagnosis
5 approaches available-
1. Microscopic demonstration of inclusion/
elementary bodies
2. Isolation of chlamydia
3. Demonstration of chlamydial antigen
4. Demonstration of antibodies
5. hypersensitivity tests
 Lab diagnosis
1. M/E of EB or IB-
• Chlamydial EB or IB are large & seen under
Light microscope
• Scrapings
• Smears
• Stained by Giemsa, lugols I2, microIF,
(Castaneda, Machiavello or Giminez stains)
 Lab diagnosis
2. Isolation of Chlamydia
Done by inoculation into
• embryonated eggs,
• animals (mice),
• tissue cultures
Embryonated egg inoculation
• Chlamydia can grow in yolk sac of 6-8 day old
chick embryos
• EB, IB can be demonstrated in yolk sac
Mouse inoculation-
• Mouse inoculation no longer in use for
chlamydial isolation
 Lab diagnosis
Cell culture-
• Preferred method for isolation
• Commonly used cell lines are McCoy (mouse
fibroblast-like), HeLa 229 (derived from human
cervical carcinoma), BHK-21 (derived from
baby hamster kidney),
 Lab diagnosis
3. Demonstration of chlamydial antigen
a. Micro IF
• Smears from ocular, genital samples are
Stained with fluorescent conjugated Ab &
examined under microscope.
b. ELISA
c .Molecular methods
DNA probes, PCR
 Lab diagnosis
4. Demonstration of Antibody in sera
• CFT or micro IF
• CFT used mainly in psittacosis &LGV
• 1:64 titre is diagnostic
• Micro IF in TRIC infections Titre of 1:8 or more
is diagnostic
 Lab diagnosis
5. Demonstration of hypersensitivity skin test
• Frei’s test for LGV
• Antigen made from bubo pus produce a
reaction in patients with LGV when injected
intradermally
 Frei’s test
• Inject antigen id in
forearm with control on
other arm
• Induration of 7mm or
more in 2-5 days was
considered positive.
 Chlamydia Trachomatis
Leading cause of ocular & genital infections
worldwide
• Trachoma
• Inclusion conjunctivitis (neonates & adults)
• Infant pneumonia
• Genital chlamydiasis
• LGV (lymphogranuloma venereum)
Chlamydia Trachomatis
 TRACHOMA
TRACHOMA
• Chronic keratoconjunctivitis
Characterized by follicles,
papillary hyperplasia, pannus
formation & cicatrisation
• Name trachoma referring to
roughness of conjunctiva
(inner surface of the eyelids)
 TRACHOMA
• leading cause of infectious blindness in the
world.
• transmission from eye-eye by fingers/
fomites/flies.
• IP variable & Onset is gradual
• C/F - Eye discharge, swollen eyelids, trichiasis
(turned-in eyelashes), swelling of lymph nodes
in front of the ears
 Trachoma Grading
1. McCallan's classification-McCallan in 1908 divided the clinical course of trachoma into 4 stages

Stage 1 (Incipient Stage 2 (Established Stage 3 (Cicatrising Stage 4 (Healed

trachoma) trachoma) trachoma) trachoma)
Hyperaemia of Appearance of mature Scarring of palpebral Disease is cured or is
palpebral conjunctiva follicle & papillae conjunctiva not markable

Progressive corneal Scars are easily visible Sequelae to

Immature follicle pannus as white bands cicatrisation cause
symptoms

2. WHO classification-The World Health Organization recommends a simplified grading system for trachoma.[4] The
Simplified WHO Grading System is summarized below:
• Trachomatous inflammation, follicular (TF) – Five or more follicles of >0.5 mm on the upper tarsal conjunctiva
• Trachomatous inflammation, intense (TI) – Papillary hypertrophy and inflammatory thickening of the upper
tarsal conjunctiva obscuring more than half the deep tarsal vessels
• Trachomatous scarring (TS) - Presence of scarring in tarsal conjunctiva.
• Trachomatous trichiasis (TT) – At least one ingrown eyelash touching the globe, or evidence of epilation
(eyelash removal)
• Corneal opacity (CO) – Corneal opacity blurring part of the pupil margin
 Lab diagnosis
Microscopy
Characteristic inclusion body Halberstaedter
Prowazek (HP bodies) demonstrated in
conjunctival scrapings after staining with
• Giemsa/ Castaneda/ Machiavello methods
• posses glycogen matrix so stained wd Lugols I2
• Fluorescent ab method(Micro IF)
Isolation
• Embryonated eggs &Tissue culture
Ag/Ab detection
 Treatment & Prevention
• Surgery to correct advanced stages of the
disease
• Antibiotics Zithromax (azithromycin)
• Facial cleanliness
• improved sanitation.
 Inclusion Conjunctivitis
neonatal form of IC
• infant gets infection during birth in birth
passage
• Infection appear between 5-12 days after birth
• benign & self limiting
• Tt is local application of antibiotics
 Inclusion Conjunctivitis
In adult form
• ch/by follicular hypertrophy with scanty non
purulent discharge.
• Known as ‘swimming pool conjunctivitis’ as
Infection occurs while taking bath in
community swimming pools that gets
contaminated with chlamydia from genital
secretions of infected person
 Infant Pneumonia
• C.trachomatis affect usually around 4-16
weeks of age
• develop prominent respiratory symptoms
with cough & wheezing
 Genital Infections
• C.trachomatis causes 2 types of genital
infection
a. Genital Chlamydiasis by occulogenital
serotype D-K
b. LGV by serotypes L1, L2 L3.
 Genital Infections
• In men – urethritis (non gonococcal),
epididymitis, proctitis, & Reiter’s syndrome
(triad of recurrent conjunctivitis- polyarthritis-
urethritis associated with many infections.
• In women- acute urethritis, Bartholinitis,
mucopurulent cervicitis, endometritis,
salpingitis, PID, conjunctivitis, & Reiter’s
syndrome
 Lymphogranuloma Venereum
• Also k/a Lymphogranuloma inguinale (STD)
• Caused by LGV serovar of C.trachomatis= L1,
L2, L3
• LGV serovars Preferred site is regional LN
• TRIC serovars preferred site is epithelial cells
 Lymphogranuloma Venereum
Primary stage
• small painless papulo-vesicular lesion on external
genitals (IP of 3 days to 5 weeks)
Secondary stage
• 2 weeks later lymphatic spread occur
• Ch/ supparative inguinal adenitis
• LN enlarge, suppurate, become adherent to skin
& pus discharging sinuses forms
 Lymphogranuloma Venereum
Tertiary stage
• Chronic, last for several years
• scarring & lymphatic blockage
 Lab diagnosis
Microscopy
Smears of material aspirated from bubos
prepared
EB (Miyagawa’s
granulocorpuscles)
 Lab diagnosis
Isolation
• by intracerebral inoculation into mice,yolk sac
of eggs,cell cultures
Ag/Ab detection
• LGV patients develop high titres of circulating ab
of 1:64 or more in CFT. Titre of 1:512 or more in
micro IF test
Skin test-intradermal Frei test done
• Treatment- Tetracycline orally for 3 weeks
 Chlamydia Psittaci
• Psittacosis is a disease
of parrots (psittacos
means parrot) & other
psitticine birds
• Transmissible to
humans
 Chlamydia Psittaci

• Chlamydia shed in droppings/ nasal discharge

• Human infection mostly occupational (poultry
workers, pigeon farmers, pet shop owners,
bird fanciers, vetenerarians)
• Route of infection inhalation
• IP= 10 days
 Chlamydia Psittaci
C/F
• Mild influenza like syndrome Fatal
pneumonia
• Infected cells (alveolar macrophages) from
patients, mouse brain, yolk sac & cell culture
show IB (Levinthal–Cole-Lillie or LCL bodies)
 Chlamydia Pneumoniae
• designated it as
C.psittaci strain TWAR
(from Taiwan Acute
Respiratory)
 Chlamydia Pneumoniae
• Common cause of respiratory disease in older
children & adults worldwide
• Clinical spectrum include- pharyngitis,
sinusitis, bronchitis & pneumonia
THANK YOU…