Atypical bacteria

Objectives
• Describe general structure, Pathogenesis and lab
findings of Chlamydiae
• Describe general structure, Pathogenesis and lab
findings of Rickettsiae
• Describe general structure, Pathogenesis and lab
findings of Mycoplasmas
• Describe general structure, Pathogenesis and lab
findings of Spirochetes
Reference:
Jawetz, Melnick & Adelberg s MEDICAL
MICROBIOLOGY 25the Edition
 Chlamydiae
-Are obligate intracellular organisms
↓
Grow only within cells
-Including 
Chlamydia trachomatis  eye, respiratory and genital tract
infections.
Chlamydia pneumoniae  atypical pneumonia
Chlamydia psittaci  psittacosis
-They lack the ability to produce sufficient energy to go
independently.
-They have a rigid cell wall (but not a typical peptidoglycan
layer)
-Have a replicative cycle different from that of all bacteria
begin when extra cellular, metabolically inert (elementary
body)
↓
Reorganize into larger metabolically active (reticulate body)
↓
release daughter repeated binary fission
elementary bodies  appear as inclusions
useful in diagnosis
-All chlamydiae share a group-specific lipopolysacharide
antigen (detected by CFT)
-They also possess species-specific and immunotype-specific
antigens (proteins) (detected by IF)
-C.psittaci and C.pneumoniae each have 1 immunotype,
where as C.trachomatic has at least 15.
Transmission
C.trachomatis infects only humans and
transmitted by close personal contact
(sexually or by passage through the birth
canal)
Individuals with asymptomatic genital tract
infections are important reservoir.
In trachoma, the microorganisms is transmitted
by finger-to eye contact.
C.pneumoniae  human  by aerosol
C.psittaci  birds  dry feces of bird 
humans
Laboratory diagnosis
Cytoplasmic inclusion seen with Giemsa's stain or
by immunofluorescence.
Organisms can be identified in exudates by
fluorescent antibody staining, ELISA, or by use of
DNA probe (hybridyzation).
Chlamydiae can be grown in cell culture or
embryonated egg
Chlamydial antigens can be detected by ELISA.
Serologic tests reliable in diagnosis of chlamydial
infection but not for C.trachomatis because the
frequency of infection is so high that many
people already have antibodies.
Rickettsiae : They are the agents of:
Typhus  R.prowazekii
R.typhi
R.tsutsugamoshi
Rocky Mountain spotted fever  R.rickettsii
Q fever  Coxiella burnetii
Are very short rods, their cell wall resemble that of Gram's
negative but stained poorly.
Are obligate intracellular parasites
Rickettsiae devide by binary fission inside host cell but by a
distinctive intracellular cycle.
Several rickettsiae (R.prowazekii, tsutsugamoshi & richettsii)
posses antigens that cross-react with antigens of the OX
strains of Proteus vulgaris
 The Weil-Felix test  anti-richettsial antibodies in serum
will agglutinate Proteus vulgaris.
Pathogenesis
Transmitted to human by bite of arthropod,
except C.burnetti which is transmitted by
aerosol.
All rickettsial infections are zoonotic only for
epidemic typhus.
The typical lesion caused by rickettsiae is a
vasculitis particularly in the endothelial lining.
Damage of vessels of skin  characteristic rash,
edema and hemorrhage.
Endotoxin is involved in pathogenesis but no
exotoxin or cytolytic enzymes.
Laboratory diagnosis
Based on serologic analysis rather than isolation
of the organism
Rickettsiae can be grown in cell culture or
embryonated egg but hazardous.
CFT provides more specific data.
Weil-Felix reaction (microagglutination test)
based on cross reaction of antigens in rickittsial
disease and with O antigen (polysaccharide)
found in Proteus vulgaris strains OX-2, O-19 & OX-
k. but the test is of limited value.
Mycoplasmas
Are a group of small, wall-less organisms of which
Mycoplasma pneumoniae is the major pathogen.
Mycoplasma pneumoniae  atypical pneumonia
Are smallest free-living organisms ( 0.3/m)
Absence of a cell wall stain poorly with Gram's
stain. And antibiotics that inhibit cell wall
synthesis are ineffective.
Pleomorphic  a flexible 3-layer cell membrane
Mycoplasmas can grow on artificial media
(including several lipids)  grow slowly (1week)
 visible colony "fried egg" shape.
Pathogenesis & Epidemiology
Only human pathogen transmitted by respiratory
droplets.
In the lungs, the organism is rod-shaped with a
tapered tip that contain specific proteins attached
to the respiratory epithelium. The mucosa not
invaded but ciliary motion is inhibited  necrosis
of epithelium.
M.pneumonia has one serotype but immunity is
incomplete, auto-antibodies produced during
infection (cold agglutinins) against red blood cells
and brain, lung and liver cells  extra pulmonary
manifestations.
Laboratory Diagnosis
Culturing sputum sample may take at least 1 week
and need special media.
Serological test  cold agglutinin titer  1:128 
recent infection
IgM auto antibodies against type O RBC 
agglutination at 4oC
(Non specific ) false positive  influenza virus
_ adenovirus
Infection can be confirmed by 4-fold rise in specific
antibody titer in CFT.
 Classification of Spirochetes

• Human pathogens belong to following 3 genera:

1. Treponema
2. Leptospira
3. Borrelia

• Others (saprophytes) are found in water, sewage

and in mouth & genital tracts of humans.
 Treponema

• Main treponemes are:

T. pallidum - Syphilis:Venereal (sexual) disease, enter
the body through minute abrasions on mucosa or skin
T. pertenue - Yaws Non venereal
T. carateum - Pinta disease

• All three species are morphologically identical

 Characteristics of T.pallidum

• Elongated, motile (endoflagella- axial filament), spiral

bacteria with tapering ends seen under Dark Ground
Microscope by staining with silver salts
• Culture –
1. Do not grow on artificial media.
2. Virulent strains can be maintained in rabbit testis by
serial passages e.g. Nichol’s strain – used for
diagnosis & research
3. Reiter’s strain – nonpathogenic strain cultivated for
diagnosis.
 Characteristics of T.pallidum
• T pallidum may remain motile for 3–6 days at 25°C. In
whole blood or plasma stored at 4°C, organisms remain
viable for at least 24 hours
• Very delicate, easily killed by drying or heating at 42C for
an hr.
• Antigenicity –has hyaluronidase
Induce Ab as
1. Non specific - Reagin Ab (cardiolipin extracted from
normal mammalian )
2. Specific – Trponemal Ab, immobilizing and killing live
motile T pallidum and fixing complement
 Laboratory Diagnosis
Specimens include tissue fluid expressed from early surface
lesions for demonstration of spirochetes; blood serum for
serologic tests.
• Dark-Field Examination The exudate is examined under oil
immersion with dark-field illumination for typical motile
spirochetes.
• Immunofluorescence Tissue fluid or exudate is fixed,
stained with a fluorescein-labeled antitreponeme serum,
and examined by means of immunofluorescence
microscopy for typical fluorescent spirochetes.
 Nontreponemal tests

• Test for reagin Ab using cardiolipin Ag.

1. VDRL (Venereal Disease Research Laboratory) test
2. RPR (Rapid Plasma Reagin) test
• used as screening tests for syphilis.
• Reagin is a mixture of IgM and IgG antibodies reactive with
the cardiolipin-cholesterol-lecithin complex.
• false positive reactions are seen in Influenza, carditis and
connective tissue diseases
 Treponemal antibody tests

• Measure Ab against T pallidum antigens. The tests

are confirmatory for positive result from a
nontreponemal test, include:
1. T pallidum hemagglutination (TPHA)
2. T pallidum immobilization (TPI) test
3. Treponemal antibody tests using the EIA for T pallidum
4. Fluorescent treponemal antibody absorbed (FTA-ABS) test
 Summary
 Chlamydiae are obligate intracellular organism including Chlamydia
trachomatis, Chlamydia pneumoniae and Chlamydia psittaci
 All share a group-specific lipopolysacharide antigenand species-specific
and immunotypes.
 Diagnosed by Cytoplasmic inclusion seen with Giemsa's stain or by
immunofluorescence, ELISA, or by use of DNA probe.

 Rickettsiae are obligate intracellular parasites devided by binary fission

inside host cell.
 All rickettsial infections are zoonotic only for epidemic typhus, the typical
lesion is a vasculitis.
 Laboratory diagnosis based on serologic analysis rather than isolation of
the organism
 Summary
 Mycoplasma are a group of small, wall-less organisms of which
Mycoplasma pneumoniae is the major pathogen atypical pneumonia.
 It is only human pathogen transmitted by respiratory droplets, inhibit
ciliary motion.
 Diagnosed by serological test  cold agglutinin titer  1:128  recent
infection

o Treponema pallidum are major human pathogens belong to Spirochetes

which cause syphilis disease, transmit through minute abrasions on
mucosa or skin
o Diagnosed by direct detection in tissue fluid or blood serum for serologic
tests which include:
1. Nontreponemal tests (test for reagin Ab using cardiolipin Ag)
2. Treponemal antibody tests (measure Ab against T pallidum antigens)