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MYCOPLASMAS & CELL-WALL

DEFECTIVE BACTERIA

MICROBIOLOGY LECTURE SERIES


LUZ GREGORIA LAZO-VELASCO, MD

Mycoplasmas

MOLLICUTES cell wall-free bacteria; 200 known species (16 of human origin)
Mycoplasma pneumoniae pneumonia, joint & other infections
Mycoplasma hominis postpartum fever
Ureaplasma urealyticum nongonococcal urethritis in men; lung disease in
premature infants of low birth weight
Mycoplasma genitalium urethral and other infections; smallest genome
Other members are pathogens of the respiratory and urogenital tracts and joints of
humans and animals

Mycoplasmas
Smallest organisms that can be free-living in nature and self-replicating on laboratory
media
Characteristics:
1. small mycoplasmas are 125-250 nm in size
2. highly pleomorphic (lack rigid cell wall; bounded by a triple-layered unit
membrane that contains a sterol (mycoplasmas require the addition
of serum or cholesterol to the medium to produce sterols for growth)
3. completely resistant to penicillin (lack cell wall structures at which penicillin
acts; inhibited by tetracycline or erythromycin

Mycoplasmas
Smallest organisms that can be free-living in nature and self-replicating on laboratory
media
Characteristics:
4. can reproduce in cell-free media; on agar, the center of the whole colony is
characteristically embedded beneath the surface
5. growth inhibited by specific antibody
6. have an affinity to mammalian membranes

Mycoplasmas
MORPHOLOGY & IDENTIFICATION
A. Typical Organisms
growth in fluid media gives rise to many different forms
growth on solid media consists principally of protoplasmic masses of
indefinite shape that are easily distorted
50-300 nm in diameter
B. Culture
media with serum, a metabolic substrate (glucose or urea), growth factors
(yeast extract)
incubation at 37oC for 48-96 hours

Mycoplasmas
MORPHOLOGY & IDENTIFICATION
B. Culture
Giemsa stains of centrifuged sediment show characteristic pleomorphic
structures
subculture on appropriate solid media yields minute colonies
after 2-6 days on biphasic (broth over agar) and agar medium incubated
in a Petri dish, colonies are round, with a granular surface and a
dark center typically buried in the agar

Mycoplasmas
MORPHOLOGY & IDENTIFICATION
C. Growth Characteristics
grow on cell-free media that contain lipoprotein & sterol
use glucose & urea (ureaplasmas) as source of energy
some produce peroxidases and hemolyze RBCs

Mycoplasmas
ANTIGENIC STRUCTURE
species are classified by biochemical and serologic features
complement fixation (CF) antigens of mycoplasmas are glycolipids
antigens for ELISA tests are proteins

Mycoplasmas
PATHOGENESIS
flasklike or filamentous shapes, specialized polar tip structures that mediate
adherence to host cells
interactive proteins - adhesins (P1 adhesin of M pneumoniae , MgPa adhesin of M
genitalium)
adherence-accessory proteins
attach to the surfaces of ciliated and nonciliated cells, probably through the mucosal
cell sialoglycoconjugates and sulphated glycolipids

Mycoplasmas
DIAGNOSTIC LABORATORY TESTS
A. Specimens: throat swabs, sputum, inflammatory exudates, respiratory, urethral
or genital secretions
B. Microscopic Examination: direct examination useless
C. Cultures: broth & special solid media
agar media incubated at 37oC with 5-10% CO2
M hominis- fried-egg appearance on agar
Ureaplasma sp- growth signalled by color change indicating hydrolysis of
urea

Mycoplasmas
DIAGNOSTIC LABORATORY TESTS
D. Serology CF tests, HI tests, indirect fluorescence, EIAs
E. Nucleic acid Amplification Tests
TREATMENT TETRACYCLINES, ERYTHROMYCINS

Mycoplasma pneumoniae
ATYPICAL PNEUMONIAS
PATHOGENESIS
Transmitted from person to person by infected RESPIRATORY SECRETIONS
Infection initiated by attachment of the organisms tip to a receptor on the surface
of respiratory epithelial cells; mediated by a specific adhesion protein on the
differentiated terminal structure of the organism
CLINICAL FINDINGS
generally mild
asymptomatic infection to serious pneumonitis, with occasional neurologic and
hematologic (hemolytic anemia) involvement & a variety of possible skin lesions

Mycoplasma pneumoniae
ATYPICAL PNEUMONIAS
CLINICAL FINDINGS
incubation period: 1-3 weeks
insidious onset
malaise, fever, headache, sore throat, cough (initially non-productive, occasionally
paroxysmal, later with blood-streaked sputum and chest pain)
early: patient appears moderately ill, physical signs of pulmonary consolidation
often negligible compared with the striking consolidation seen on radiographs
later: infiltration at its peak, illness may be severe
resolution of pulmonary infiltration and clinical improvement occur slowly (1-4weeks)

Mycoplasma pneumoniae
ATYPICAL PNEUMONIAS
CLINICAL FINDINGS
most common pathologic findings: interstitial with peribronchial pneumonitis and
necrotizing bronchiolitis
Common causes of community-acquired bacterial pneumonia in addition to
Mycoplasma pneumoniae:
Streptococcus pneumoniae
Legionella pneumophila

Haemophilus influenzae
Chlamydia pneumoniae

Mycoplasma pneumoniae
ATYPICAL PNEUMONIAS
LABORATORY TESTS
diagnosis of M pneumoniae pneumonia is largely made by the clinical recognition of
the syndrome
WBC count slightly elevated
sputum Gram stain does not suggest some other bacterial pathogen
Culture highly specialized test; almost never done to diagnose M pneumoniae
infection
Cold hemagglutinins for group O human erythrocytes appear in 50% of untreated
patients, in rising titer, maximum reached in the 3rd or 4th week after onset
(>1:64)

Mycoplasma pneumoniae
ATYPICAL PNEUMONIAS
LABORATORY TESTS
rise in specific antibodies to M pneumoniae demonstrable by CF tests (acute &
convalescent phase sera necessary to demonstrate a 4-fold rise in the CF
antibodies
EIA detect IgM and IgG can be highly sensitive and specific; more sensitive than CF
tests
PCR assays of specimens from throat swabs or other clinical material can be
diagnostic

Mycoplasma pneumoniae
ATYPICAL PNEUMONIAS
TREATMENT
Tetracyclines, macrolides, fluoroquinolones can produce clinical improvement but
do not eradicate M pneumoniae

Mycoplasma hominis
Strongly associated with infections of the uterine tubes (salpingitis) and tubo-ovarian
abscesses

Ureaplasma urealyticum
Requires 10% urea for growth
Causes nongonococcal, nonchlamydial urethritis in men (biovar 2)
Common in female genital tract
Associated with lung disease in premature LBW infants who acquired the organism
during birth

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