MYCOPLASMA

infections
Dr.T.V.Rao MD
Edmond Nocard and Emile Roux
(1898)
 Nocard and Roux contributed for
Mycoplasma discovery
●
In 1896 Nocard and Roux reported the
cultivation of the causative agent of
contagious bovine pleuropneumonia
(CBPP), which was at that time a grave and
widespread disease in cattle herds. The work
of Nocard and Roux represented the first
isolation of a mycoplasma species.
 History
●
The name Mycoplasma, from the Greek
mykes (fungus) and plasma (formed), was
proposed in the 1950s, replacing the term
pleuro pneumonia-like organisms (PPLO)
referring to organisms similar to the causative
agent of CBPP. It was later found that the
fungus-like growth pattern of M. mycoides is
unique to that species.
 Mycoplasma
●
Mycoplasma species
are the smallest free-
living organisms.
These organisms are
unique among
prokaryotes in that
they lack a cell wall.
 Overview of Mycoplasma Infections
●
Mycoplasma species are the smallest free-
living organisms and are unique among
prokaryotes in that they lack a cell wall. This
feature is largely responsible for their biologic
properties, including lack of a Gram stain
reaction and nonsusceptibility to many
commonly prescribed antimicrobial agents,
including beta-lactams. Mycoplasma
organisms are usually associated with mucosa.
 Can part of Normal flora
●
They reside extracellularly in the respiratory
and urogenital tracts and rarely penetrate the
sub mucosa, except in the case of
immunosuppression or instrumentation, when
they may invade the bloodstream and
disseminate to numerous organs and tissues.
 Important Human Mycoplasma
infecting Human
●
Family – Mycoplasmataceae – requiring
cholesterol or other sterols as an essential
grwoth factor.
a. Genus Mycoplasma which utilize glucose
or arginine but donot split urea.
b. Genus Ureaplasma – which hydrolyze
urea
 Basic Characters of Mycoplasma
●
Prokaryotic microbes
●
Size of 150-250 nm
●
Lack of a cell wall
●
Sterol-containing cell
membrane
●
Fastidious growth
requirements
●
Fried-egg or mulberry
colonies on agar
Mycoplasma are cell wall deficient
microorganisms
●
Cross-section of
Mycoplasma
bacteria, a common
cause of atypical
pneumonia. This
bacteria is unusual
in that it lacks a cell
wall.
 Culturing Mycoplasma
●
Mycoplasma can be
cultured on liquid or solid
medium
●
Growths optimally at 35 to
370c
●
Medium of growth should
be enriched with 20% horse
or human serum.
●
The colonies appears as
fried egg appearance
 Characters of Mycoplasma
●
They are prokaryotes but lack a cell wall.
However, they have a unique cell membrane
that contains sterols, which are not present in
either bacteria or viruses. Mycoplasma
organisms are small (150-250 nm) and have
deformable membranes. The name
Mycoplasma refers to the plasticity of the
bacterial forms resembling fungal elements.
 Important Species in Mycoplasma
●
Scientists have isolated at least 17 species of
Mycoplasma from humans, 4 types of
organisms are responsible for most clinically
significant infections that may come to the
attention of practicing physicians. These
species are Mycoplasma pneumoniae,
Mycoplasma hominis, Mycoplasma
genitalium, and Ureaplasma species
How they differ from other Bacteria
●
They have sterols in the cell membrane.
●
They share no DNA homology with known
bacteria.
●
They have low guanine levels plus cytosine
content.
●
Their genome has a low molecular weight.
●
They exhibit no reversion to walled forms.
 How Mycoplasma differ from other
common bacteria.
●
However, the term mycoplasma has been frequently
used to denote any species included in the class
Mollicutes). The common characteristics are the
complete lack of a bacterial cell wall, osmotic
fragility, colony shape and filterability through 450-
nm pore diameter membrane filters. The relatively
close Phylogenetic relationship of these genera was
measured by comparative sequence analysis of the
5S and 16S ribosomal RNA (rRNA)
 Specific characters of Mycoplasma
●
However, the term mycoplasma has been frequently used to
denote any species included in the class Mollicutes). The
common characteristics are the complete lack of a bacterial
cell wall, osmotic fragility, colony shape and filterability
through 450-nm pore diameter membrane filters. The
relatively close Phylogenetic relationship of these genera was
measured by comparative sequence analysis of the 5S and
16S ribosomal RNA (rRNA). The rRNA sequence analyses
also revealed that the Mollicutes are not at the root of the
bacterial Phylogenetic tree, but rather developed by
degenerate evolution from gram-positive bacteria with a low
mol% G+C (guanine plus cytosine) content of DNA,
 How they differ from Viruses
●
They grow on cell-free
media in vitro.
●
They contain both
RNA and DNA.
●
They have both
intracellular and
extracellular parasitism
in vivo.
Genomic structure is documented
 How Race Influce Mycoplasma
Infections
●
Patients with sickle cell disease or related
hemoglobinopathies are at increased risk for severe
M pneumoniae infections and may develop large
pleural effusions and marked respiratory distress.
Since sickle cell disease and other related
hemoglobinopathies are most common among
African Americans, severe complications of
mycoplasmal infections also occur most frequently
in this group of patients.
 Mycoplasma found on surfaces of
Mucous Membranes
●
Mycoplasma are found most often on the
surfaces of mucous membranes. They can
cause chronic inflammatory diseases of the
respiratory system, urogenital tract, and joints.
The most common human illnesses caused by
Mycoplasma are due to infection with M.
pneumoniae, which is responsible for 10-20%
of all pneumonias.
 Antigenic properties
●
The surface antigens
are glycolipids and
protiens
●
Glycolipids are
identified by
complement fixation.
●
Proteins antigens
detected by ELISA
method.
 Resistance
●
They are normally destroyed by heat at 450c
in 15 minutes.
●
They are relatively resistant to pencillins, and
Cephalosporins
●
Sensitive to Tetracyclnes, and several other
antibiotics
 Why Mycoplasma are Pathogenic
●
The lack of a cell wall most probably facilitates the
close contact between M. pneumoniae and its host
cell and guarantees the exchange of compounds,
which support the growth of the bacterium. As a
consequence of this bacterial surface-parasitism the
host cell is severely damaged. The exchange of toxic
metabolic compounds is discussed as a possible
cause of cell damage, however, at this stage not a
single toxic compound has been identified as a
causative agent of cell damage.
 Spread of Mycoplasma Infections
●
The disease is world wide, and
found in all age groups,
●
Transmission by drop let
infection of nasopharyngeal
secretions.
●
Spread is associated with close
contact of infected person
●
Important infection in Military
personal.
●
Even the persons recovered
from infection will harbor the
pathogens for 2 moths or more
 Clinical Manifestations

●
Generalized aches and pains
●
Fever (usually 102°F)
●
Cough - Usually non-productive
●
Sore throat (nonexudative Pharyngitis)
●
Headache/ myalgias
●
Chills but not rigors
●
Nasal congestion with coryza
●
Earache
●
General malaise
 Respiratory spread
●
Infection moves easily
among people in close
contact because it is
spread primarily when
infected droplets from
the respiratory system
circulate in the air due
to coughing, spitting, or
sneezing
Pneumonia leading Manifestation in
Mycoplasma infections
Pneumonia
●
Pneumonia caused
by Mycoplasma is
also called atypical
pneumonia, walking
pneumonia, or
community-acquired
pneumonia.
 Mycoplasma Pneumonia

Mycoplasma pneumonia is most often seen in

children and young people. Up to 15 % of all
cases of pneumonia in patients younger than
40 years are caused by mycoplasma
pneumoniae. Most mycoplasma infections are
manifested clinically as bronchitis and/or
Pharyngitis. Pneumonia develops in between 3
and 10% of the patients.
Mycoplasma presents as non specific
Respiratory infections
●
Infections commonly
involve the oropharynx,
trachea, bronchi, and lungs,
usually causing unilateral
pneumonia of the lower
lobe. The
radiographic appearance
cannot be distinguished
from that of other
nonbacterial pneumonias.
 Mycoplasma contributes to several
Respiratory infections
●
M pneumoniae infections lead to clinically apparent
disease involving the upper respiratory tract. In 5-
10% of patients (with the rate depending on age), the
infection progresses to tracheobronchitis or
pneumonia and is usually self-limited. Pleural
effusion (usually small) occurs in 5-20% of patients.
Mycoplasmas have also been implicated in the
pathogenesis of asthma, leading to acute and chronic
wheezing in some individuals.
 Radiological presentation
●
The radiological picture is
extremely variable, but one
or both lower lobes are
usually involved. The
opacities usually start as
partly mottled, partly node-
like peribronchial opacities,
which may gradually
develop to involve whole
segments or lobes
 Other Systemic Infections in
Mycoplasma
●
In rare instances other organs may be
involved (central nervous system,
pancreas, joints, skin, heart, and
pericardium), probably as a result of
haematogenous spread.
 Mycoplasma in children
●
In children younger than 3 years primarily develop
upper respiratory infection.
●
M pneumoniae infection is uncommon in the first
year of life; however in neonates, it may cause severe
disease.
●
M pneumoniae infection is common in school-aged
children, with the highest rate of infection in
individuals aged 5-20 years, in whom the tendency is
to develop bronchitis and pneumonia.
 Mycoplasma in New born

●
Colonization of infants by
genital Mycoplasma species
usually occurs during
passage through an infected
birth canal, and genital
mycoplasmal organisms
have been isolated from the
upper respiratory tract in
15% of infants.
Colonization usually does
not persist beyond 2 years.
 Cardiac Manifestations

●
Arrhythmia and/or
ECG abnormalities
(conduction defects)
●
Congestive failure
●
Pericarditis
●
Myocarditis
●
Endocarditis
●
Dr.T.V.Rao MD
 Neurological Manifestations
●
Cranial neuropathy
●
Aseptic meningitis or Meingoencephalitis
●
Transverse myelitis
●
Brainstem dysfunction
●
Dysfunction of the pyramidal or extrapyramidal tract
●
Cerebellar dysfunction
●
Cerebral infarction
●
Guillain-Barré syndrome
●
Peripheral neuropathy

Dr.T.V.Rao MD
 Musculoskeletal Manifestations

●
Polyarthralgias
●
Acute arthritis
(monoarticular or
migratory)
●
Digital necrosis
Dr.T.V.Rao MD
Ureaplasma species
 Important species in Ureaplasma
●
The Ureaplasma genus now is subdivided into
2 species: U urealyticum and U parvum. For
clinical purposes, separating infections caused
by the different 2 species is not possible or
necessary. In both the clinical setting and in
the diagnostic laboratory, they are considered
Ureaplasma species.
Ureaplasma differs from Mycoplasma
●
The Ureaplasma are the
only non fermentative
mollicutes i.e., they do not
ferment the growth
substrates such as
carbohydrates and amino
acids like other mollicutes
but they depend on the
hydrolysis of urea for their
energy
Urease test differentiates Mycoplasma
from Ureaplasma species
 UREAPLASMA UREALYTICUM
●
Some strains of Mycoplasma frequently isolated
from Urogenital tract of human beings and animals
●
They are also called T strains or T form mycoplasma
●
They are peculiar to hydrolyze urea, which is
essential grwoth factor in addition to Cholesterol
●
At present the have been reclassified as Ureaplasma
urealyticum.
Ureaplasma can be a Normal flora in
sexually active individuals
●
U. urealyticum is part of the normal genital flora of
both men and women. It is found in about 70% of
sexually active humans.
●
It had also been described to be associated with a
number of diseases in humans, including
non-specific urethritis (NSU), infertility,
chorioamnionitis, stillbirth, premature birth, and, in
the perinatal period, pneumonia,
bronchopulmonary displasia[1] and meningitis.
 Spread of Ureaplasma Infections
●
Major infections are
produced by
M.hominis and
M.urealyticum
Commonly spread
by Sexaul
contact
 Ureaplasma can be opportunistic
pathogen in pregnancy
●
Ureaplasma urealyticum, a common
commensals of the urogenital tract of sexually
mature humans, is gaining recognition as an
important opportunistic pathogen during
pregnancy. While its etiologic significance in
many aspects of adverse pregnancy remains
controversial, recent evidence indicates that
U. urealyticum in the absence of other
organisms is a cause of chorioamnionitis.
 Ureaplasma can produce several
disseminated complications
●
Evidence indicates that U. urealyticum is a cause of
septicaemia, meningitis, and pneumonia in newborn
infants, particularly those born prematurely. There is
strong but not definitive evidence that Ureaplasma
infection of the lower respiratory tract can lead to
development of chronic lung disease in very low-
birth-weight infants. Although risk factors for
colonization of the lower genitourinary tract have
been identified,
 Ureaplasma species: causes
●
Urethritis
●
Pyelonephritis
●
Pelvic inflammatory disease
●
Endometritis or chorioamnionitis
●
Infectious arthritis
●
Surgical wound infections
●
Neonatal pneumonia
●
Neonatal meningitis

Dr.T.V.Rao MD
 Mycoplasma and sterility
●
Mycoplasma species do not cause vaginitis, but they
may proliferate in patients with bacterial vaginosis
and may contribute to the condition. M hominis has
been isolated from the endometria and fallopian
tubes of approximately 10% of women with
salphingits; M genitalium may also be involved in
pelvic inflammatory disease and Cervicitis. Whether
Ureaplasma infection causes involuntary infertility
remains speculative.
Mycoplasma in HIV infections
●
Mycoplasmas tend to cause
more severe infections in
the HIV infected persons
and cause prolonged
infections.
●
Other Immunosupressed
patients are susceptible to
Mycoplasma infections
 Diagnosis in Immunosupressed a
priority
●
The correct
microbiological
diagnosis takes on
greater importance in
patients who are
Immunosuppressed and
at greater risk for
disseminated infection
and a poor outcome.

Dr.T.V.Rao MD
DIAGNOSIS Mycoplasma
infections
Majority of cases are Diagnosed with
Serological Tests
 Cold Agglutination Test
●
Cold Agglutination test
is associated with
macroglobulin
antibodies that
agglutinate human o
RBC at low
temperature
Cold Agglutination test procedure
●
The serial dilutions of patients
serum are mixed with an equal
volume of 0.2% washed human
O group erythrocytes at low
temperature
●
The clumping is observed at 40c
overnight.
●
However the clumping is
dissociated at 370c
●
A titer of 1:32 or > is suggestive.
●
A raised titer in paired serum
sample is more suggestive of
infection.
RBC showing non agglutinating and agglutinating
RBC
 Streptococcal MG test
●
The test is performed
by mixing serial
dilutions of patients
serum with heat killed
suspension of
Streptococcus MG.
●
The sample is
incubated at 370c
●
The agglutination titer
of 1:20 or > is
suggestive.
 Other Serological Tests
●
Immunofluorescence
●
Hemagglutination
inhibition test
●
Complement fixation
test less sensitivie.
Growth of Bacteriological Medium
●
For isolation swabs from
throat or respiratory
secretions inoculated not
Mycoplasma medium
●
The growth is slow and
takes 1 – 3 weeks
●
The colonies appear as
fried egg, with central
opaque granular area
surrounded by flat
translucent peripheral zone
Mycoplasma on PPLO agar
 Typical Mycoplasma colonies on
enriched medium
●
The colonies showing
typical fried egg
appearance.
●
The colonies appear 2-6
days of incubation.
●
The size of the colonies can
be from 10 – 600 microns
in size.

Dr.T.V.Rao MD
Colony characters of Mycoplasma
isolates
 M. pneumoniae colonies demonostrated
in Dienes method
●
The colonies can be
demonostrated by Dienes
method.
●
In which a block of agar
containing the colony is cut
and placed on a slide,
covered with a cover slip
on which has been dried in
alcoholic solution of
methylene blue and azure.
Biochemical Characters of
Mycoplasma
●
The metabolism of
Mycoplasma are
fermentative
●
Most species utilize
glucose or arginine
●
Urea is hydrolyzed by
Ureaplasma only
 Diagnosis of Urogenital Infections
●
Material from urethra, cervical, or vaginal or
centrifuged deposit of urine is added to
separate vials with liquid mycoplasmal
medium containing phenol red and 0.1%
glucose, arginine or urea
●
The Ureaplasmal urease also breaks down
urea to ammonia
 Newer methods in Diagnosis
●
Phylogeny based rapid
identification of
urogenital
Mycoplasmas and
ureaplasmas based on
amplification of part of
165rRNA gene by PCR
is available

Dr.T.V.Rao MD
Emerging trends in PCR primers
●
Now several DNA
primers specific for
several Mycoplasmas
are available and useful
for specific
identification of species
 Advantages of PCR methods
●
PCR methods
are proving to
be rapid,
sensitive, and
specific
Drugs used in Mycoplasma infections
●
Erythromycin (M. pneumoniae and
Ureaplasma spp.)
●
Tetracyclnes.
●
Clindamycin (M. hominis)
●
Levofloxacin
●
Doxycycline
●
Gentamycin
Created for benefit of Medical
and Paramedical students in
Devloping world
Dr.T.V.Rao MD
Email
doctortvrao@gmail.com