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MYCOBACTERIA

Dr. Sudheer Kher


Classification of
Mycobacteria
1. Tubercle bacilli 1. Atypical Mycobacteria
(Runyon Groups)
a) Human – MTB
b) Bovine – M. bovis
a) Photochromogens
b) Scotochromogens
3. Lepra bacilli c) Nonphotochromoge
a) Human – M. ns
leprae d) Rapid growers
Tuberculosis
(TB, Consumption, Koch’s Disease)
• M. tuberculosis
• major human disease
– healthy people
• problems
– association with AIDS
– multiple drug-resistance
–Chronic disease
–Prolonged treatment
General characters of the
genus
 Slender rods
 Resist staining but once stained,
resist decolorization by dilute mineral
acids; hence called ACID FAST
BACILLI (AFB)
 Aerobic, Non-motile, Non-sporing,
Non-capsulated.
 Growth generally slow
Mycobacterium
tuberculosis
 One of the most serious infectious
diseases in the developing world
 One third of world’s population infected
with M. tuberculosis
 Thirty million people have active disease
 Nine million new cases occur
 Three million people die of the disease,
each year.
Mycobacterium tuberculosis
(MTB)
 Morphology –
– Ziehl – Neelsen stain – Once stained by
Carbol fuchsin, resist decolorization by
20% Sulphuric acide and absolute
alcohol. Acid & Alcohol Fast (AFB)
– Fluorescent dyes like Auramine O or
Rhodamine also stain and the
decolorization is resisted.
MTB : Cultural characters
 Grow slowly. Generation time
14-15 hrs
 Colonies appear after 2
weeks or at 6-8 weeks
 MTB - Obligate aerobe
wenstein Jensen Medium –

lective. Always in screw capped bottle. Bluish Green.


ntains – Egg protein – Solidifying agent
Mineral salts – Mg sulphate, Mg citrate
Asparagine
Malachite Green – Selective agent
erilized by - Inspissation
Mycobacterium tuberculosis
(MTB)
 Morphology –
– Straight or slightly
curved rods
Modes of infection

1- Droplet infection
Person to person by inhalation aerosols
Mycobacterium tuberculosis (Pulmonary
tuberculosis)

2- Ingestion of milk
Infected cattle
Mycobacterium bovis (Intestinal tuberculosis)

3- Contamination of abrasion
Laboratory workers (Skin infection)
Pathogenesis of
tuberculosis
• infects lung

• distributed within macrophages

• facultative intracellular pathogen


– inhibits phagosome-lysosome fusion
– resists lysosomal enzymes
Tuberculosis
Clinical picture :
* Low grade fever
* Weight loss
* Night sweats
* Fatigue
* Cough & haemoptysis
Laboratory Diagnosis
 Demonstration of bacilli
 Culture & isolation or Animal
inoculation
 Demonstration of hypersensitivity to
tubercular protein
 Serological tests limited value
Specimen
* According to site of infection :
- Sputum - Urine - Body
fluids
- Gastric lavage - Blood - Tissue
biopsy

* Specimens need appropriate processing

Liquefaction with N-acetyl-L-


cysteine
Sputum Decontamination with NaOH
Laboratory Diagnosis
 Pulmonary TB –
– Specimen –
 Sputum – Early morning, if scanty 24 hrs, three
consecutive day samples. Laryngeal swabs or gastric
lavage in children.
– Microscopy –See at least 100 field / 10
minutes.
 Grading –
– 1+ -> 3-9 bacilli in entire smear
– 2+ -> 10 or more in entire smear
– 3+ -> 10 or more bacilli seen in most oil
immersion fields
Laboratory diagnosis
M. tuberculosis
 Acid fast bacteria in sputum
 Culture on L J media
 Biochemical identification
 Antibiotic sensitivity test
 Tuberculin test
 PCR
Laboratory diagnosis -

tuberculosis
• skin testing
– delayed hypersensitivity
– tuberculin
– protein purified
derivative, PPD
• X-ray
Tuberculin Test (Mantoux
test)
 Delayed hypersensitivity
skin test to assay:
cell mediated immunity
to tubercle bacillius
Material: A purified
protein derivative (PPD)
Dose : 0.1 ml of (PPD)
is injected intradermal
Reading : Positive test is
defined as
- Induration equal or
greater than 10 mm
- Develop 48-72 hours
after injection
Positive skin test
-tuberculosis
• indicates exposure to
organism
• does not indicate active
disease
Laboratory Diagnosis
 Extra -Pulmonary TB –
– Specimen –
 CSF – in suspected meningitis
 Pleural fluid & other exudates
 2-3 days urine in renal TB
 Biopsy material.
Treatment
 Chemoprophylaxis – INH for one year
 Domicilliary treatment preferred
 Drugs –
– Rifampicin
– Isoniazide Bactericidal
– Pyrazinamide
– Streptomycin
– Ethambutol
– Ethionamide
– Thiacetazone Bacteriostatic
– Paraminosalicylic acid
– Cycloserine
Treatment
 Short term chemotherapy of six
months is sufficient
 Problem area – Development of
resistance by mutant selection
– Solution – Treatment by two to three
drug combination, adequate treatment.
Immuno-prophylaxis

 Intradermal injection of live


attenuated vaccine Bacille Calmette-
Guerin (BCG).
 The strain causes self limited lesion
and induces hypersensitivity &
immunity.
 Coverts tuberculin negative person
to positive reactor.
 Immunity lasts for 10-15 years.
Immunity 60-80%
BCG
 Given at birth without tuberculin
testing
 Protects against TB, the disease runs
milder course in protected, prevents
skeletal, meningeal & miliary forms.
 Also found useful in leprosy,
leukaemias and other malignancies
by non-specific stimulation of RE
system.
Mycobacteria

Mycobacterium leprae
Classification of Atypical Mycobacteria
Ridley Jopling Classification of Leprosy
Basis - Immunological

RR-Reversal reaction; ENL-Erythema nodosum leprosum; PB-Paucibacillary, MB-


Multibacillary