MYCOPLASMA

Introduction
• Mycoplasma is a genus of bacteria that lack a cell wall
around their cell membrane.
• without a cell wall,they are unaffected by many common
antibiotics such as penicillin or other beta-lactam
antibiotics that target cell wall synthesis.
• They can be parasitics or saprotropic.
• Several species are pathogenic in human,includinng
M.pneumoniae, which is a important cause of atypical
pneumonia and other respiratory disorder.
 History
• Louis Pasture first noticed mycoplasm while observing the
causitive agent of pleuropneumonia in culture.
• Mycoplasma was first reported by Nocard and Roux.In the
year 1930s,it become evident that mycoplasma cannot be
defined as viruses.

• They were first isolated from pleural fluid of cattles

suffering from bovine pleuropneumonia that’s why it is
known as PPLO [pleuropneumonia like organism].
 Classification
• Kingdom :Bacteria
• Phyluum:Firmicutes
• Class:Mollicutes
• Order:Mycoplasmataless
• Family:Mycoplasmataceae
• Genus:Mycoplasma
• SPECIES:
• M.myocides
• M.genitalism
M.hominis
M.hyopneumoniae
 Morphology of mycoplasma
• Gram negative bacilli
• Approximatelly size:250-300nm
• Shape:pleomorphic but more or less flask shape
• Shows glinding motility
• lack cell wall
• Can pass through bacterial filter
• No flagella
• Non sporing
Structure
Culture charecteristics of mycoplasma
• Aerobes and facultatives anaerobes.
• Optimum temprature 35-37℃.
• Optimum ph 7.4
• Required 20 ％ human or horse seum and yeast
extract to grow thus they are fastidious in nature.
 Most common used culture media
1. PPLO broth containing glucose,penicillin,
phenol red
2. Hayflick modified medium containing heart
infusion broth
 Mycoplasma inoculated in broth and incubated
at 37℃ for 3-10 days or sometimes even upto
1-3 weeks produce the uniform turbidity and a
color change on medium from orange to yellow
indicator was reported due to fermentation of
glucose.
 PPLO Agar with penicillin,
20 ％ human or horse serum
containing media
• mycoplasma inoculated on plate and incubated for 2-6
days at 37 produce 10-600micrometer fried egg like
apperence colony.
 Biochemical test
• Suger fermentation test : ferments glucose but not
maltose lactose with the production of acid but not gas.
• Arginine hydrolysis test: positive
• Urease fermentation test: negative
 Antigenic structure
• Membrane glycolipids and protein are the major
determinants of mycoplasma.
 membrane glycolipids shows cross
reaction with human tissue and other
bacteria.
 many antigenically distinct species of
mycoplasma have been isolated from
animals [eg.mice,chicken,turkeys].
 antigene of mycoplasma can be
identified by complemente fixation
test and ELISA.
 Pathogenesis
• Mycoplasm is known to cause the distinct type of
infection varying from pneumonia,non gonococcal
urethritis,uterine tube infection,post partum fever,joint and
other infection.
• Source of infection
1. Normal commensal of mycoplasma species found in
human body [endogenous].
2. Mycoplasma found in nature or animal [exogenous].
• Mode of transmission
 mostly endogenous
 exogenous route
• mycoplasma attach to the surface of ciliated and non
ciliated cell,probably through the adherence
protein,interactive protein [HMW1,2] which is found in
polar tip structure of mycoplasma.
• They necrotized or kill the cell[infected] by distinct
mechanism,mention below:
1. direct toxicity through the generation of hydrogen
peroxide and super radicals OH.
2. Cytolysis mediated by ag-ab reaction.
3. chemotoxis and action of mononuclear cells.
4. completion for deplection of nutrient.
• Clinical manifestation
1. Pulmonary infection
 upper respiratory tract infection
  Lower respiratory tact infection
1. Extra pulmonary infection
 Neurological infection
 dermalogical infection
 cardiac infection
 Rheumatologic infection
 hematologic infection
 Genito-urinary tract infection
a. Upper respiratory tact infection
i. pharyngitis
ii. trachea bronchitis
iii. sore throat(rarely)
iv. otitis media
a. Lower respiratory tact infection
 atypical pneumonia (M.pneumonia) also called eatan agent
pneumonia,primary typical or walking pneumonia.it is
characterized by: fever,
headchae,malaise,chills,coughing,peribronchal pneumonia.
1. Extra pulmonary infection
 Neurological infection
-meningoencephalitis
-Guillain barre syndrome
-aspectic meningitis
 Dermatological infection
-formation of large number of erythematous in skin
rashes all over the body.
 Cardiovascular infection
-mycarditis,pericarditis,endocarditis etc.
 Genitourinary tract infection
-caused by urogenitalmycoplasma species such
as ;M.hominis,M.fermentans,M.genitalium,ureoplasm spp.
-they frequently colonize in female lower tract urogenital
tract infection such as ; vagina, periurethal area,cervix etc.
-microorganism is transmited mostly by sexual contact or
mother to fetus during birth.
 Hematological infection
-anemia, hypercoagulopathy.
 Laboratory diagnosis
1) Clinical specimen;throat swab,nasopharyngeal
aspirates,lung biopsies,sputum,urine,urethral swabor
urethral dicharge.
2) Storage and transport;
• clinical specimen should be immediately placed into
following transport media to avoid dry.
• standard mycoplasm fluid medium containing fetal
bovine serum,gelatin and penicillin.
• viral transport medium,added with ampicillin and
cefotaxime.
• transport should be immediate,if delay in transport the
specimen should be stored at 4℃for 48 hr .
3.Microscopic examination
• Electron microscope:mycoplasma appears as flasked
shaped structure.
• gram staining: gram negative bacilli
• giemsa staining:red or pink color bacilli.
4.Ag detection test
• Direct immunofluorescent test
 used for detecting p1 adhesion ag of M.pneumonia.
 smear from distinct clinical specimen such as
nasopharyngeal swab, sputum are stained withantiP1
monoclonal antibody tagged with fluorochrome dye and
observed under the fluoresent microscope.
 mycoplasma present in clinical specimen were fluoresent
green against blue background.
 5.Culture media used
• PPLO broth and agar
• serum is necessary for growth
• in broth, growth indicated by ph change due to
carbohydrate metabolism.
• colonies best seen under microscope Dienes staining
technique.
• may take 2-3 weeks.
6.Boichemical test
• Suger fermentation test : ferments glucose but not maltose
lactose with the production of acid but not gas.
• Arginine hydrolysis test: positive
• Urease fermentation test: negative
7.Colonies examination
• size varies 200-250micrometer for mycoplasma and 15-
30micrometer for ureaplasma colonies are examined by:
1.hends lens
2.Dienes staining
• mycoplasma pneumonia formed in culture plate, black of
agar conatining mycoplasma colonies is cut and
transferred to glass slide and flooded with alcoholic
solution of methylene blue and azure examined under low
power of microscope.
• mycoplasma retain color for atleast 2days and appear
intense royal blue where as ureoplasma appear reddish to
greenish blue.
 8.Colonies of mycoplasma may be identified by
1.Hemadsorption test
• M.pneumonia agglutinates guinea pig red blood cells
(RBS) and the colonies on agar absorb RBC to their
surface and become red.
2.Tetrazolium reduction test
• colonies of M.pneumoniae appear red when bacterial
colonies are flooded with solution of tetrazolium
compound to red color formazan by mycoplasma
pneumonia bacteria.
9.Serological test
a) Specific ab detection test
• Ab specific to M.pneumoniae and glycolipid ag can be
detected after about weeks of infection and peak at 3-6
weeks then decline gradually.
• IgM ab increases in children, IgA in adults .test like ELISA
,later agglutination test, immuno fluorescence aasay.
a) Non specific test
• mycoplasma posses certain heterophile ag such as surface
glycolipid haptens that cross react with1ag of RBC or
carbohydrate ag of group F streptococcus cell wall.
• this property can be used to detect heterophile ab in patient
sera by using non specific ag tes eg:cold agglutination test
etc
 10.Prevention
• chemoprophylaxis of mycoplasma infection is not recommended and
no vaccine is available.
11.Treatment
• Macrolides are drug of choice (oral azithromycin)
• Alternative drugs are
• Doxycycline
• levofloxacin
• Moxifloxacin etc.