Assignment no:1 DIT Neha Gosai

1. The picture is of Mycobacterium tuberculosis colonies.

2. Mycobacterium tuberculosis is the cause of tuberculosis in most patients.
3. It is a long and thin acid-fast rod.
4. It is a strict aerobe and is not referred to as gram-positive or gram-negative
because its acid-fast nature is much more relevant in a clinical setting.
5. The Löwenstein–Jensen medium, more commonly known as LJ medium, is
a growth medium specially used for culture of Mycobacterium species,
especially Mycobacterium tuberculosis.
6. When grown on LJ medium, M. tuberculosis appears as brown, granular
colonies (sometimes called "buff, rough and tough").
7. The prefix Myco may make you think of fungi, but this is a bacterium. The
prefix in the name came from the mistaken impression that colonies growing
on agar resembled fungal colonies.
 8. It grows very slowly, with a generation time of 15 to 20 hours.
9. A period of up to 6 weeks is required for colonies to appear in culture.
10.Robert Koch identified that M. tuberculosis often forms serpentine cords
while growing, and he called the unknown substance causing this style of
growth “cord factor.”
11.Cord factor appears to be associated with virulent strains, and it is a lipid
component of the mycobacterial cell wall.
12.All mycobacterial species have walls that have a very high content of
complex lipids, including mycolic acid and waxes.
13.This chemical characteristic makes them relatively impermeable to stains
and difficult to decolorize (acid-fast) once they are stained.
14.The lipid wall of the bacterium also influences its virulence and makes it
resistant to drying and disinfectants.

Composition of LJ medium for M.tuberculosis

 Malachite green
 Glycerol
 Asparagine
 Potato starch
 Coagulated eggs
 Mineral salt solution
 Potassium dihydrogen phosphate
 Magnesium sulfate
 Sodium citrate
 The original formulation included starch, which was later found to be
unnecessary, so omitted.
 Low levels of penicillin and nalidixic acid are also present in LJ medium to
inhibit growth of Gram-positive and Gram-negative bacteria, to limit growth
to Mycobacterium species only. Presence of malachite green in the medium
inhibits most other bacteria. It is disinfected and solidified by a process of
inspissation. Presence of glycerol enhances the growth of M. tuberculosis.
 If the slopes are made on test tubes, they must be stored in cold and used
within a month.
 For cultivation of M.bovis, glycerol is omitted and sodium pyruvate is
added.
 The medium appears green, opaque, and opalescent.