Chapter 25

Mycoplasma and Ureaplasma

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Significant Human Pathogens

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 Mycoplasma
 Two mycoplasma species are known human
pathogens.
 Mycoplasma pneumoniae
• Respiratory disease
 Mycoplasma hominis
• Urogenital tract disease
 Mycoplasma species are the smallest self-
replicating organisms in nature.
 Very difficult to detect
 Common contaminant of cell culture
• Important in research and in virus labs utilizing cell culture

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 General Characteristics of
Mycoplasma
 Do not possess cell walls
 Sometimes referred to as CWD (cell wall deficient)
 Resistant to cell wall active antibiotics
 Penicillins, cephalosporins
 Bonus is that antibiotics can help reduce normal florae
 Slow growing
 Fastidious
 Require cholesterol and fatty acids for growth
• Note these are conditions found in cultures growing
eukaryotic cells
• Human sites

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 General Characteristics of
Mycoplasma (Cont.)
 Mycoplasma
 Sometimes known as pleuropneumonia-like
organism (PPLO)
• Eaton agent: from discoverer
 Colonies grow with center imbedded below agar
surface
• Thus appear as “fried eggs”
 Transmission
 Direct sexual contact, during delivery, respiratory
secretions, or fomites
• Very susceptible to heat and drying conditions

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 General Characteristics of
Mycoplasma (Cont.)
 Location of infection
 Epithelium of mucosal surfaces in respiratory and
urogenital tracts
 Adhere tightly to epithelial cells
 Oropharynx
• Relatively uncommon for M. hominis but common for
M. pneumoniae
 Urogenital tract
• M. hominis (~10%-50%) women

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Human Pathogens in the
Class Mollicutes

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Mycoplasma Illustrating “Fried
Egg” Appearance

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Electron Micrographs of
M. pneumoniae

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Electron Micrographs of
M. pneumoniae (Cont.)

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Mycoplasma Species Indigenous
to Humans

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 Clinical Infections of
M. pneumoniae
 Diseases
 Bronchitis
 Pharyngitis
 Walking pneumonia (primary atypical pneumonia)
 Mostly asymptomatic (over 90%)
 Isolation always considered significant
 20% pneumonia in general populations
• School-age children and young adults
 50% in confined settings
• Prisoners, college students, and military personnel

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Chest Radiograph of Atypical
Pneumonia

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Summary of Genital Mycoplasmas with
Urogenital and Newborn Diseases

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 Clinical Infections of
M. pneumoniae (Cont.)
 Spread via close contact
 Aerosol droplets
 Incubation
 2 to 3 weeks
 Headache, low-grade fever, malaise, anorexia,
sore throat, dry cough, earaches
 Extrapulmonary complications
• Cardiovascular, central nervous system (CNS),
dematologic, and gastrointestinal problems are rare.

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 Clinical Infections of
Mycoplasma hominis
 Infections of the lower urogenital tract
 Found in 50% of healthy patients
 Can cause infections of the upper urinary tract in
sexually active people
 Opportunistic pathogen
• May cause more severe infections in immunocompromised
 Salpingitis: inflammation of the fallopian tubes
 Pyelonephritis: infection of kidney and ducts
 Pelvic inflammatory disease (PID)
 Postpartum fevers

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 Clinical Infections of
Ureaplasma urealyticum
 Infections of the urogenital tract
 Normal florae of the lower urinary tract of women
• Still can cause upper urinary tract infections
• Significant due to infection of fetus
 Chorioamnionitis (infection of placental membrane)
 Congenital pneumonia
 Chronic lung disease in premature infants
 Meningitis of newborns with negative cultures
 10% of cases of nongonococcal urethritis (NGU)
in men

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 Other Mycoplasma Species
 M. genitalium
 Has been some association with NGU, cervicitis,
endometriosis, and PID
• May lead to tubal sterility
 Very difficult to culture, takes 2 to 3 months
• Interferes with serology for other mycoplasma
 M. fermentans (incognitus)
 Likely an opportunistic respiratory pathogen
• Adults with respiratory illness
• Acute immunodeficiency syndrome (AIDS)–related mycoplasma
 Synovial fluid of patients with rheumatoid arthritis

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 Major Clinical and Diagnostic
Manifestation of M. pneumoniae

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 Specimen Collection
 Extremely sensitive to drying due to lack of
cell wall
 Swabs in transport medium
 Dacron polyester or calcium alginate
 Trypticase soy broth
 If not plated immediately, freeze specimen at
–70°C

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Flow Diagram for
Mycoplasma spp.

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Identification of Mycoplasma and
Ureaplasma
 M. pneumoniae
 Generally not cultured
• Takes too long, and sensitivity is low
 Use serology
• 2 to 4 weeks apart for fourfold rise in titer
 M. hominis and U. urealyticum
 Culture
• Initially in liquid media, and watch for pH change
 SP4, Shepard’s 10B, or 2SP broth
 Plate enriched culture, and check for characteristic
colonies

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Identification of Mycoplasma and
Ureaplasma (Cont.)
 M. hominis
 Requires arginine
• Turns pink
 Release of ammonium (NH4) from arginine (phenol red
indicator)
 Plate to agar
• A8 agar
• Look for characteristic fried egg colonies of a variety of
shapes and sizes
• Diene’s or methylene blue stain
 Light blue “egg white”
 Dark blue “yolk”

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Diene’s Stain for
Mycoplasma spp.

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Illustration of Mixed Sizes of
Mycoplasma

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Detection of Mycoplasma Using
DNA-Fluorochrome Stain

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 Identification of Mycoplasma and
Ureaplasma (Cont.)
 U. urealyticum
 Requires urea
• Turns pink
 Release of NH4 from urea
 Plate to A8 agar plate
• Look for irregularly shaped colonies
• If using U9B, color test medium
 Add magnesium chloride (MgCl2)
– Rust brown color at membrane as MgCl2 is converted to
magnesium peroxide (MgO2)

 Examine under low power on stereoscopic microscope

 T-strain Mycoplasma (tiny)

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Mixed Isolation of M. hominis and
U. urealyticum

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Comparative Features of Laboratory
Methods Detecting Mycoplasmas
and Ureaplasmas

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 Treatment
 M. pneumoniae
 Erythromycin
• Scattered reports of resistance
 Tetracycline

 M. hominis
 Resistant to erythromycin; use clindamycin or
lincomycin
 U. urealyticum
 Resistant to clindamycin or lincomycin; use
erythromycin

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 Laboratory Detection of Frequent
Respiratory Nonbacterial Pathogens

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