KEYWORDS

e PPD
Here s a how chan to help keep track of whom Mycobactenum
we are ’alking ahout.
tuberculosis

Mycobacterium bovis

Mycobacterium microt/

Mycobacterium a/r/canum

Runyon I

Mycobacterium Runyon II M.
u/ceranz Nontubercuious scrofulaceum
Mycot›acteria Mycoaacten”u
Runyon Ill m
avium complex
Runyon IV if. c/›e/onei and
f
Mycobacterium leprae
General characters of the
genus
Slender rods Aerobic, Non-motile,
Resist staining but Non-sporing, Non-
once stained, resist Capsulated.
decolorization by Growth generally slow
dilute mineral acidst Genus includes
hence called — Obligateparasites
(AFB — Opportunist pathogens
) — Saprophytes
Mycobacterium tuberculosis
• One of the most serious infectious diseases in

• One third of wo Id s population infected with

C.

- three million people die of the disease, each

Morphology
—
— Ziehl — Neelsen stain — Once stained by Carbol
fuchsin, resist decolorization by 20o/oSulphuric acide
and absolute alcohol. Acid & Alcohol Fast (AFB)
— Fluorescent dyes like Auramine O or Rhodamine also
stain and the decolorization is resisted.
— Reason for Acid & Alcohol Vastness —
Presence of unsaponifiable wax Mycolic acid
Semi permeable membrane around the cell
Property of cell wall and related to integrity of the cell wall
Staining may be uniform or granular
 MTB : Cultural characters
• Grow slowly. • MTB grows more
Generation time luxuriantly (eugonic)
14-15 hrs than N. bovis
(dysgonic).
Colonies Addition of 0.5%
appear after 2 Glycerol supports
weeks or at 6-8 growth of
weeks
MTB - Obligate human
strains. No effect or
aerobe inhibitory effect on
bovine strains.
Solidmedia Liquid media
—Egg containing ——
Dubos
—
— Proskauer & Beck s
Middlebrooks
Petragnini medium
— Sula s
Dorset
— Blood containing —
— Tarshi s Sauton s
— Serum containing
— Loeffle s serum slope
— Potato containing
— Pawlowsky s
 Resistance
• Not specifically resistant to heat. 60 C x 20 min
destroys.
In sputum can survive 20-30 hrs
Relatively resistant to disinfectants. Survives
exposure to
— 5 % Phenol
15 % Sulphuric acid
— 3 % Nitric acid
—5% Oxalic acid
—4 o N ã S H
Biochemical
reactions
Biochemical
reactions
Modes of infection

Mycobacterium tuberculosis (Pulmonary

tuberculosis)

Mycobacte ium bov s (intestinal tuberculosis)

Pathogenesis of
tuberculosis
 .i
Cel mediated
tuberculosis
Classical tubercular lesion — Granuloma with typical
Langhan s giant cells, epithelioid cells, lymphocytes
and fibrosis.
There are multiple light areas (opacities) of varying size that
run together (coalesce). Arrows indicate the location of
cavities within these light areas. The appearance is typical
for chronic pulmonary tuberculosis.
 Tuberculosis
Clinical picture :
Laborato diagnosis
Selective. Always in screw capped bottle. Bluish Green.
Contains — Egg protein — Solidifying agent
Mineral salts— Mg sulphate Mg bitrate
Asparagine
Malachite Green— Selective agent
Ster‹lized by - Inspissation
La boratory
Tu bercu Test test)
Positive skin test -
tuberculosis
 Tubercu in Test
' A positive test indicates p evious exposure and car iage of T.B.
” A negafi e tubercuł in Lest excludes infection in suspected pc sors
' Tuberrulin positive persons may develop eaütivation ty ß Ot T.ò.
Tuberculin negati e persons are at risk Of gaining new infection

- Sever tuberculosis infection (Milia y T.ß.) - Hodgkin s

disease

Child en below 5 years of age with no exposure history

 Demonstrationof
• Culture
bacilli & isolation or Animal inoculation

tubercular protein
Serological tests limited value
” According to site of infection :
- Sputum - Body fluids
Urine - Tissue biobsy
- Gastric lavage
Blood
” Specimens need appropriate processing

Sputum Decontamination with NaOH

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
- fields
2 -> 10 or more in entire smear
3 -> 10 or more bacilli seen
— Culture &oil immersion
in most
—
ASTAnimal inoculation — Animal of choice Guinea
Pig.
Extra -Pulmonary TB
—
— Specimen—
CSF — in suspected meningitis
Pleural fluid & other exudates
2-3 days urine in renal TB
Biopsy material.
Purpose — Homogenization &
Concentration in sputum & other
specimens.
Methods —
• Useful for microscopy, culture & animal
inoculation
— Petroff s method
MOSt Wldely used
Equal volumes of Sputum 4o/oNaOH incubated at 37C X
min.Centrifuge
20 at 3000 rpm X 30 min. Sediment neutralized
by N/10 HCI.
Can be used for smear culture, animal inoculation
— Simpler method
To avoid cOntriŁugatiOn & NeUtralizatiOn
Equal volumes of sputum + solution of Cetrimoniom bromide
20 g + NaOH 40 g in one litre. Allow to stand for five
minutes. Inoculate Acid Buffered LV Pledium with swab.
Laboratory diagnosis
(cu re)
 Pathogenesis
” Inhalation of tubercle bacilli

”They multiply in the alveolar macrophages

” An early tubercle (granuloma) is formed

 Pathogenesis
” Lesions, healing or progression of infection
depend upon
1 Dose of infecting mycobacteria
2 Resistance and hypersensitivity of host
” Virulence :
Glycolipids on the outer surface of bacteria
- Enhance granuloma
formation
- Inhibit migration of polymorphnuclear
 Pathogenesis
An exudative lesion :
- spread to regional lymph nodes
- A scar of healing may later calcify (Ghon
FOCUS)
- Lymph nodes caseate and then calcify
- Bacilli in the lesion slowly die
Tuberculin test becomes positive
- The person immune & hypersensitive
 Pathogenesis
* Activation of tubercle bacilli due to immunity
” Formation of tubercles that caseate
- fibrosis
-open into a bronchus (open tuberculosis) ”
Tube cle bacilli erode a blood vessels
- Infect any organ (biliary T.B.)
 Treatment
Chemoprophylaxis — INH for one year
Domicilliary treatment preferred
Drugs —
Rifampicin
Ison‹az de Bactericidal
Pyrazinam de
Str eptomycin
‹ •. •
• ..'
 Treatment
• Short term chemotherapy of six months is
sufficient
Problem area — Developmentof
resistance
by
—mutant
Solution selection
— Treatment by two to three drug
combination, adequate treatment.
 Treatment

Four drugs are usually started in initial therapy due to:

- Caseous material blocks penetration of drugs

- Some bacilli persist in a metabolically inactive state
 Treatment

2- Second line drugs (more toxic and less effective):

Noncompliance (failure to complete the course}:

 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
60-800/o
 BCG
» Given at birth without tuberculin
testing
• Protects
milder course
againstinTB,
protected, prevents
the disease runs
skeletal, meningeal & miliary forms.
Also found useful in leprosy, leukaemias
and other malignancies by non-specific
stimulation of RE system.
 Copyrig I t 20D5 by Randy Glasbergen. www.gtasbergen.co m

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