Mycoplasmas

Hugh B Fackrell

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 Presentation Outline
Structure
Classification
Multiplication
Clinical manifestations
Epidemiology
Diagnosis
Control

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 Pleuropneumonia
organism
The mycoplasmas are essentially
bacteria lacking a rigid cell wall during
their entire life cycle, although they
are also much smaller than bacteria.
The first organism of this type was
associated with pleuropneumonia of
cattle, and was originally called the
pleuropneumonia organism (PPO).

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 General Characteristics
smallest known free-living organisms.
Because of the absence of cell walls, they do
not stain with the Gram stain, and they are
more pleomorphic and plastic than eubacteria.
Giemsa stain
they appear as tiny pleomorphic cocci, short rods,
short spirals, and sometimes as hollow ring forms.
Their diameter ranges from 0.15 u to 0.30 u.

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 Mycoplasma, Ureaplasma
very small (0.2 x 0.8 um)
pass through a 0.45 um filter
No Cell wall: plasma membrane only
resistant to antibiotics that interfere
with the integrity of cell wall;
penicillins, cephalosporins,
vancomycin, bacitracin
susceptible to tetracycline, erythromycin

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 Structure
The cell is enclosed by a limiting membrane
which is more similar to that of animal cells
than that of bacterial cells because of sterols
present in the membrane.
The cytoplasm contains ribosomes,but lacks
mesosomes. There is no nuclear membrane.
In some strains, amorphous material on the
outer surface of the membrane suggests the
existence of a capsule.

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 Mycoplasma
requires sterols for growth, can be
grown on laboratory media
most are facultatively anaerobic
Exception M. pneumoniae
replication controversial
replication time 1-6 hours

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 Mycoplasma pneumoniae
AKA Eatons agent
aerobic but very slow growing
extracellular pathogen: attaches to
respiratory epithelium by an attachment
factor called P1
interacts with a glycoprotein receptor on the
epithelial cell surface
ciliostasis is followed by epithelial cell
destruction

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 Clinical Syndrome
Pneumonia
walking pneumonia frequently
confused with virus infection
primary atypical
clinical
Tracheobronchitis
Pharyngitis
differential diagnosis from Strep
throat

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 Children most susceptible

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 No Seasonal Incidence

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 Urethritis
1/2 of urethral infections not
caused by Chlamydia or N.
gonorrhoeae.
Caused by
Mycoplasma hominus
Ureaplasma

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 Infection of Tracheal ring
Organ culture

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 Destruction of host
due to release of hydrogen
peroxide and superoxide anion.

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 Laboratory diagnosis
Culture:
fried egg colonies on medium containing
sterols
Most mycoplasmas require a rich medium
containing a sterol and serum proteins for
growth.
Serology:
Complement Fixation test,
Hemagglutination

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 Laboratory Diagnosis
Culture Mycoplasma from sputum,
mucous membrane swabbings or
other specimens
direct inoculation into liquid or
solid media containing serum,
yeast extract and penicillin to
inhibit contaminating bacteria.

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 Cultural Characteristics
Despite the lack of a cell wall, they do not
require a medium of very high osmotic
pressure.
On solid media, they form minute,
transparent colonies.
looks like a fried egg. The different strains vary in
their growth rate
may take from two days to several weeks to
form a colony.

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 Fried Egg Colonies

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 Fried Egg Colonies
Stain intensely with
neutral red or tetrazolium
or methylene blue.

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 fixation
on acute and convalescent serum.
patients serum heated to 56C to eliminate
complement
combine patients serum and known
Mycoplasma antigen in presence of added
complement. Mix.
Incubate - add indicator system
Red cells and anti-red cell antibody
hemolysis occurs if complement is unused.

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 Hemagglutination
Cold agglutinins to human O
erythrocytes.
hemabsorption & B-hemolysis of
guinea pig red blood cells.

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 Identification
conclusively identified by staining
its colonies with fluorescein-
labelled antibody.

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 M. pneumoniae Nucleic
Acid Probes
specific recombinants to
oligonucleotide sequences that are
only found in Mycoplasma
pneumoniae.

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 L Forms
Some bacteria readily give rise
spontaneously to variants that can replicate
in the form of small filterable protoplasmic
elements with defective or absent cell walls.
These organisms, called L-forms, can also
be formed by many species when cell wall
synthesis is impaired by antibiotic
treatment or high salt concentration.

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 L Forms vs Mycoplasma
contain a rigid cell wall, at least at
one stage of their life cycle
no sterols in their cytoplasmic
membrane.

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 Pleuropneumonia-like
organisms
Several organisms with similar
morphological characteristics and
cultural properties have been isolated.
These are commonly referred to as
pleuropneumonia-like organisms or
PPLO. A certain group of mycoplasmas
produce extremely tiny colonies on agar
plates, and are called the T-strains.

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 Metabolism
The parasitic mycoplasmas have
truncated respiratory systems,
lacking quinones and cytochromes.
Another indication for the simplicity of
the electron transport chain is the
finding that the reduced nicotinamide
adenine dinucleotide (NADH) oxidase
activity is cytoplasmic.

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Arginine dihydrolase Pathway
pathway Complex electron transport
chains are usually membrane bound, since
they depend on the spatial organization of
their components. Ruling out oxidative
phosphorylation as an ATP-generating
system leaves only two proven ways of ATP
generation, both based on substrate level
phosphorylation. The major source for ATP
is the arginine dihydrolase pathway.

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 Metabolism
A few species derive their energy from the
degradation of glucose or the hydrolysis of
urea.
All species synthesize DNA, RNA, lipids and
proteins.
Not known if they can synthesize amino acids.
Those species that require sterols incorporate
these sterols (mainly cholesterol) into the cell
membrane up to concentrations of 65%.

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 Multiplication
In the absence of a rigid cell wall, the
pattern of replication is quite different
from that of typical bacteria, whose
division starts with the formation of a
well-defined septum.

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 Life Cycle of PPLO
Elementary
body

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 Fragmentation of filaments
mechanism of division in
mycoplasmas is controversial,
sequential microscopic observation
suggests that new elementary
particles arise by fragmentation of
filamentous cells containing
several discrete DNA components.

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 DONE!!!

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