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Lectures-5-6-Mycobacterium Tuberculosis, M. Leprae, Actinomycetes
Lectures-5-6-Mycobacterium Tuberculosis, M. Leprae, Actinomycetes
GENUS: MYCOBACTERIUM
Prof. Khalifa Sifaw Ghenghesh
Obligate aerobe, Gram-positive rods
Acid fast
Complex cell wall lipids
– include mycolic acids
– protects vs. phagolysosomal components
Peptidoglycan, glycolipids
– acid-fastness
Two major groups:
– Slow growers:
– Rapid growers:
Mycrobacterium tuberculosis
Mycobacterium bovis:
Lowenstein-Jensen Plate Culture Inoculated
with 15 Strains of Mycobacterium Species
CULTURE CHARACTERISTICS
On primary isolation:
– visible growth after up to 8 weeks
Colonies:
– Buff colour, dry bread crumb-like appearance
– Growth is eugonic (M. bovis = dysgonic)
Growth temperature:
– 35-37oC
Obligate aerobe
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Heat-sensitive
Susceptible to alcohol, glutaraldehyde and
formaldehyde.
Some differential characteristics of
tuberculle bacilli causing human disease
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Species Atmospheric
preference Nitratase TCH Pyrazinamide
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M. tuberculosis Aerobic + S S
M. bovis Microaerophilic -- R R
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TCH = thiophen-2-carboxylic acid hydrazide
S = sensitive, R = resistant
THE DISEASE
Not highly contagious:
– transmission with prolonged contact
between susceptible and active case
– usually transmitted by airborne droplets,
must penetrate deep into respiratory tree
– infection can be via other routes:
ingestion=> infection through cervical
or mesenteric LN
Virulence
– Ability to Survive within Macrophages
Primary Tuberculosis
Post-Primary Tuberculosis
Stages of Primary Tuberculosis in Childhood
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Stage Time (from onset) Characteristics
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1. 3-8 weeks Primary complex (PC) develops and
tuberculin conversion occurs
2. 2-6 months Progressive healing of PC, possibility of
pleural effusion
3 6-12 months Possibility of miliary or meningeal
tuberculosis
4 1-3 years Possibility of bone or joint tuberculosis
5 3-5 years Possibilty of genito-urinary or chronic skin
tuberculosis
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Main differences between primary and post-primary
tuberculosis in the non-immunocompromised patients
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Characteristics Primary Post-primary
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Local lesion Small Large
Lymphatic involvement Yes Minimal
Cavity formation Rare Frequent
Tuberculin reactivity Negative (initially) Positive
Infectivity Uncommon Usual
(pulmonary cases)
Site Any part of lung Apical region
Local spread Uncommon Frequent
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TUBERCULIN TEST
Tuberculin: a heat-concentrated filtrate of a
broth in which tubercle bacilli had been grown.
Injection of tuberculin into the skin >>
– Large, indurated reactions >>Post-Primary
Tuberculosis.
– No induration >> Protective immunity
Purified Protein Derivatives (PPD):
– Mantoux Method (Intracutaneous)
– Heaf Method (Spring-loaded gun)
– Tine Tests (Disposable single tests)
Mycobacteria-Positive PPD
LABORATOY DIAGNOSIS
1. Specimen:
– Pulmonary tuberculosis: > Sputum, Bronchial
washings, Laryngeal swabs, and Early-
morning gastric aspirates.
– Homogenized tissue biopsies.
– Examine after centrifugation: Deposits of
CSF, Pleural fluid, Urine and other fluids
2. Microscopy:
– Ziehl-Neelsen Stain
– Fluorescent dyes
3. Culture:
– Decontamination:
– Lowenstein Jensen medium
Reducing Overcrowding
Vaccination:
– Bacille Calmette-Guerin (BCG)
– Not effective as control measure
Mycobacterium leprae
Leprosy (Hansen's disease)
– A chronic intracellular infectious disease
unique to man (with few exceptions).
Usually not fatal.
Never been cultivated in vitro
Armadillos:
– 1010 bacilli/gram of diseased tissue
M. leprae can be grown in mouse foot pads
or the nine-banded armadillo (picture).
PATHOGENESIS
Schwan cell >> Nerve damage >>
Anaesthesia and Muscle paralysis >>
Gradual destruction of extremities
> Nasal bones and eyes
ACTINOMYCETES
Gram +ve, Filaments that Break Up
Into Bacillary and Coccoid Forms.
i. Direct Examination:
Sputum, Pus, etc.. >> Examined
for Granules
ii. Culture: Brain Heart Infusion Agar
TREATMENT
– Penicillin >> Several Weeks
Sulfur granule from human actinomycosis tissue
section (hematoxylin and eosin stain).
2. NOCARDIA SPECIES
N. asteroides
– Nocardiosis.
Aerobic.
Disease Begins as Pulmonary Infection >
> 50% of Patients are Immunocompromised.
Fatality Rate >>
LABORATORY DIAGNOSIS
i. Direct Examination:
Sputum, Skin Lesions, Tissue Biopsies or Surgical Material
>> Microscopically.
Observe: G+ve, Multiple Branched and Beaded Filaments.
> Partially Acid-Fast.
ii. Culture:
iii. Identification: Biochemically.
TREATMENT
Sulphonamides, NA, TMP-SMX.
Nocardia asteroides
Silver stain showing the twisted masses of
long filamentous organisms
Nocardia asteroides
Acid fast stain shows the pale red staining
organisms in an area of necrosis
Differences between the genera
Actinomyces and Nocardia
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Actinomyces species Nocardia species
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Facultative anaerobes Strict aerobes
Grow at 35-37oC Wide temp range of growth
Oral commensals Environmental saprophytes
Non-acid-fast mycelia Usually weakly acid-fast
Endogenous cause of Exogenous cause of disease
Disease
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