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TB Medium Base acc.

Medium proposed by LÖWENSTEIN (1931) and modified by JENSEN (1932) for the detection and testing the resistance
of tubercle bacilli.
TB Medium Base acc. to LÖWENSTEIN-JENSEN

This culture medium complies with recommendations of the Bull. Literature

Int. (1954) and the DIN-Norm 58943. Bull. Intern. Un. Tuberc., 24; 102 (1954).

Typical Composition (g/1.6 litre) CRUCKSHANK, R.A., STEWART, S.M.: Detection of resistance to
streptomycin PAS and isoniazid in tubercle bacilli (Assoc. of Clin.
Potassium dihydrogen phosphate 2.5; magnesium sulfate Pathologists, broadsheet No. 32, 1961).
heptahydrate 0.24; tri-magnesium dicitrate 14-hydrate 0.6; DIN Deutsches Institut für Normung e.V.: Tuberkulosediagnostik,
L-asparagine 3.6; potato meal 30.0; malachite green 0.4. Modifiziertes Löwenstein-Jensen-Kulturmedium für Anzüchtung von
Also to be added: Tuberkulosebakterien - DIN 58943 .
JENSEN, K.A.: Reinzüchtung und Typenbestimmung von Tuberkelbazillen-
Glycerol 12 ml; whole-egg homogenate 1 litre.
stämmen. - Zbl. Bakt. I. Orig., 125; 222-239 (1932).
Preparation LÖWENSTEIN, E.: Die Züchtung der Tuberkelbazillen aus dem strömenden
Blute. - Zbl. Bakt. I. Orig., 120; 127-129 (1931).
Suspend 37.5 g in 0.6 litre demin. water, if required add 12 ml
glycerol, mix, autoclave (15 min at 121 °C). Cool to about 50 °C, Ordering Information
add 1 litre whole-egg homogenate prepared from fresh hen eggs
under sterile conditions; stir to give a homogeneous mixture Product Merck Cat. No. Pack size
avoiding formation of bubbles. Dispense into sterile test tubes TB Medium Base acc. to 1.05400.0500 500 g
and allow to coagulate in a slant position by heating for LÖWENSTEIN-JENSEN
45minutes at 85 °C in an inspissator saturated with water
vapour or in free-flowing steam. The culture medium should be Glycerol (about 87 %) 1.04094.0500 500 ml
heated once more in this way after about 24 hours to guarantee
its sterility.
pH: 4.8 ± 0.2 at 25 °C (before adding the homogenate). Quality control
The prepared medium is green and non-transparent.
Test strains Growth
Experimental Procedure and Evaluation Mycobacterium tuberculosis good / very good
Inoculate the culture medium massively by spreading the sample ATCC 25177
on the surface. Use the glycerol-free culture medium when Mycobacterium fortuitum ATCC6841 good / very good
culturing glycerophobic mycobacteria.
Mycobacterium kansasii ATCC 12478 good / very good
Incubation up to 2 weeks at 35 °C.
Mycobacterium phlei ATCC11758 good / very good
Check the tubes for colony growth after 10-14 days and then in
weekly intervals. The cultures should be carefully aerated. Mycobacterium smegmatis good / very good
ATCC 14468

Appearance of Colonies Microorganisms

On medium containing Mycobacterium tuberculosis
glycerol, colony growth is type humanus (R-variant)
"eugonic", i.e. abundant, raised,
crumbly, dry, usually yellowish
(navel form). This growth
pattern develops poorly on
medium which does not
contain glycerol
On medium containing type bovinus (S-variant)
glycerol, sparse growth or no
growth at all. On medium
without glycerol growth is
"dysgonic", i.e. flat, moist,
glossy, confluent colonies
(often nipple form) without
pigment formation
Rapid growth in the form of a
moist, fairly abundant "lawn":
optimal temperature 41-42 °C type poikilothermorum
optimal temperature 25 °C type gallinaceus

Merck Microbiology Manual 12th Edition 455