 Mycoplasma pneumoniae

 Mycoplasma hominis
 Mycoplasma genitalium
 Ureaplasma urealyticum
 Smallest known free-living forms (0.3 – 0.8um);
 NO CELL WALL bacteria
▪ 3 layers of cell membrane (cholesterol)
▪ Pleomorphic
▪ Resistant to antibiotics that attack the cell wall

 Belong to the class Mollicutes (soft skin in Latin) and the family of
Mycoplasmatacaea

 Mollicutes appear most closely related to bacilli, streptococci and

lactobacteria

 Most are facultative anaerobes

 Slow growing
 Highly fastidious, requiring sterols and fatty acids for membrane
fxn and growth EXCEPT for M. pneumoniae (aerobic) and M.
hominis (rapid grower).

Mollicutes
 They produce microscopic colonies
(15-300um in dm)

 Mycoplasma spp. often grow beneath

the agar

 M. hominis forms colonies w/ slightly

raised centers “fried egg
appearance”.
“fried egg appearance” of
M. hominis
 The 1st Mycoplasma was isolated in 1800s 1. M. pneumoniae – community acquired
from a cow w/ pleuropneumonia, then from pneumonia
humans; thereby the name 2. M. hominis –urogenital tract disease
3. U. urealyticum –urogenital tract disease
pleuropneumonialike organism (PPLO) or
the Eaton agent
 Pathogenesis:
 Mycoplasmas are NF of the URT and GUT ▪ colonize mucosal surfaces of URT and GUT;
▪ rarely produce invasive diseases except in
 Infants are commonly colonized w/ immunocompromised pxs;
U. urealyticum and M. hominis, but will not ▪ have great affinity to ciliated and non-
persist after 2yrs of age; on puberty ciliated epi cells
colonization results from sexual contact ▪ M. pneumoniae has P1 adhesin (causes
interaction with host cells )
 MOT: vertical, during birth, sexual contact
(M. hominis)
 MOT: contaminated URT secretions, fomites
(M. pneumoniae)
ORGANISM CLINICAL MANIFESTATIONS

Mycoplasma pneumoniae URT infection in young children

LRT infection in adults (primary atypical pneumonia/Walking
pneumonia)

Mycoplasma hominis Nongonococcal urethritis (NGU)

Salpingitis, pyelonephritis, PID (upper GUT)
Postpartum fevers

Mycoplasma genitalium Nongonococcal urethritis in men;

Cervicitis and endometritis in females

Mycoplasma fermentans Opportunistic respiratory pathogen

Mycoplasma penetrans Isolated from urine of homosexual males w/ HIV infection

Mycoplasma salivarium Isolated from synovial fluid of pxs w/ rheumatoid arthritis

ORGANISM CLINICAL MANIFESTATIONS

Ureaplasma urealyticum Nongonococcal urethritis (NGU) – in males only

Chorioamnionitis – females

Congenital pneumonia and chronic lung disease – infants

Arthritis and cystitis – hypogammaglobulinemic pxs

Ureaplasma parvum Nongonococcal urethritis (NGU) – in males only

Respiratory distress - infants

1. Specimen Collection and Transport NOTES:
2. Direct Examination
3. Culture • M. pneumoniae - attempt for
4. Isolation and Identification
isolation is not frequently done due
5. Serologic Diagnosis
6. AST to very slow growth and very
fastidious nature; serological testing
is recommended

• M. hominis and U. urealyticum are

less fastidious ( requires cholesterol)

• M. hominis – only sp that will grow

on BAP and CAP
Specimens Transport
Body fluids TSB w/ 0.5% albumin and penicillin (400units/mL)
Wound aspirates
Nasopharyngeal swab SP4 medium – sucrose PO4 buffer,
Cervical and vaginal swabs Mycoplasma base, horse serum (20%), phenol red
(Dacron polyester/Ca alginate)
Tissue

Ideally, these specimens shd be inoculated at NOTE: Sample is immediately processed or frozen
bedside or transported to the lab ASAP!!! at -70degC if plating is not done in 24 hrs.
(they lack cell wall, hence, very sensitive to
heat and drying)
 Fluorescence microscopy –
acridine orange as staining
agent

 PCR - difficulty in
interpretation (after acute
infection) since orgs are
chronically harbored by pxs Ureaplasma urealyticum
Media Indication Mollicute Requirement
SP4 broth M. pneumoniae M. pneumoniae Glucose
SP4 agar M. hominis M. genitalium
NYC agar Mycoplasma spp. M. hominis Arginine
Ureaplasma spp.
Ureaplasma spp Urea
A8 agar M. hominis pH 6.0 (Shepherd’s 10B arginine
Ureaplasma spp. broth)

 Incubation condition – anaerobic or w/ 5-10% CO2

▪ M. hominis and U. urealyticum – 1- 5 days
▪ M. pneumoniae – 1 to 2 weeks

 Ureaplasma spp are difficult to maintain in culture since:

▪ Death occurs rapidly when urea is depleted
▪ They are sensitive to pH changes during urea utilization

 Mycoplasmas do not produce turbidity in broth, phenol red is used to indicate growth
1. Colonies from A7 medium are stained with Dienes or
methylene blue. M. hominis will demonstrate a
typical “fried – egg appearance” (periphery is
light blue, center is dark blue); Most mycoplasma will
show a mixed colony presentation

2. Chen Assay – makes use of fluorochrome dye

(Hoecst 33258); highlights Mycoplasma spp as “ fried-egg” appearance of M.
hominis
small ovoid bodies in glacial HAc-fixed cell
culture

3. GP-RBC-HAD/Hemadsorption Assay -
0.5% guinea pig rbcs in PO4-buffered saline is
incubated for 20-30mins @ RT, colonies are
observed for adherence of rbcs: M. pneumoniae
(+), M. hominis (-)
mixed colony presentation
4. M. hominis colonies on CNA or ANA will appear as
pinpoint (0.05mm), clear, glistening colonies in 48 hrs.
“fried- egg” morphology is NOT observed in ANA.

M. pneumoniae M. Hominis and Urealyticum

1. Ureaplasma agar (MES) – 2-N – morpholino
ethane sulfonic acid – horse serum (provides
cholesterol), yeast dialysate, urea, phenol red, MES as buffer,
lincomycin and penicillin. Colonies will show “fried egg”
appearance and are dark-colored due to production of urease.

2. Stereomicroscope for identifying extremely small

colonies of Ureaplasma spp (T-strain mycoplasmas)

Urealyticum in culture
3. Urease activity (U9B urease color
test medium)– urease-positive colonies of
Ureaplasma are dark golden-brown due to deposition
of manganese dioxide

Urea + manganese chloride --------- manganese dioxide

urease

Urealyticum in A7 agar
1. Cold agglutinin test – cold agglutinins are non-specific antibodies produced
in response to atypical pneumonia (only 50% of pxs w/ M. pneumoniae). Cold
agglutinins are IgMs that agglutinate human grp O erythrocytes at 4degC but not
at 37degC.
▪ These Abs are detected in the px’s serum after the onset of the disease and peak during the
convalescent period.
▪ A fourfold increase in Ab titer b/n the acute and convalescent specimens is diagnostic for current
infection
▪ A titer greater than 1:28 is also diagnostic for current infection.
▪ This test, however, is no longer recommended for diagnosis of M. pneumoniae infection
due to its low specificity

2. Complement Fixation Assay

3. Immunofluorescnet antibody testing
4. Latex agglutination
5. ELISA
Cold agglutinin test
 Mollicutes are inherently resistant to beta-lactams but has remained
susceptible to:
▪ Tetracyclines
▪ Fluoroquinolones (newer)
▪ Macrolides (i.e. erythromycin)

 M. pneumoniae infections are self-limiting, no treatment required but to

shorten disease duration, the above-mentioned antibiotics can be used.

 M. hominis – resistant to erythromycin, susceptible to clindamycin and lincomycin

 U. urealyticum – resistant to clindamycin and lincomycin, susceptible to erythromycin

 Both are sensitive to tetracycline, but high level resistance is emerging