Professional Documents
Culture Documents
Common Bacteria.
Mycobacteria are mostly rod shaped, which is rectangular. They have a waxy capsule,
which when stained with Zeil Neilson shows as Red. This is an acid stain, so these are
referred to as Acid Fast Organisms (AFO). They are found inside Langahans giant cells,
as the cells have ingested and tried to kill the bacteria.
M bovis the cause of Bovine Tuberculosis, ( the reason we are here), organisms are
usually fairly rare in a TB granuloma, and are usually large rods.
M paratuberculosis, causes Johnes disease. These are smaller rods and are often present
in very large numbers forming clumps or clusters, within the giant cells. The calcification
in and around mycobacterial lesions caused by M paratb infections is often as lots of very
small areas refered to as “Fine Calcification”.
Often Johnes disease lesions can be recognized by these charactistic features.
M avium, is now considered to be more appropriately referred to as M “soil”, as these
were originally thought to be associated with birds, but now thought to be more soil. i.e.
Calf sheds. Lesions are difficult to distinguish from those of M bovis.
Parasites; eosinophils dominate the granuloma reaction, but other cells are also present.
THE FRESH SAMPLE, if required is then cultured. This involves a culture “Broth”
which is made similar to the body tissues in which the bacteria thought to be causing the
lesion are known to grow.
Mycobacteria grow very slowly and best in slightly greater CO2 levels than normal air.
This slow growing habit means other more abundant bacteria may Over-grow
mycobacteria, so antibiotics are added. The broth is cooled to solidify on a slope in the
culture tube, (Solid Culture Medium) and is then checked for growth every 30 days for
a total of 90 days. Very slow for Mr farmer to wait on!
The newer Bactec method of culturing Mycobacteria in a liquid broth can only be
achieved if no overgrowing bacteria are present. Sampling is done aseptically from the
centre of the lesion. If the lesion has been lying around for too long then other bacteria
will often invade it and so the Bactec is overgrown and we have to wait the 90 days for
the solid culture.
Fresh samples are also smeared on a microscope slide and stained. If large numbers of
AFOs are seen, then these can be “harvested” and the DNA multiplied and checked. This
is PCR (see local rules notes on PCR).
M paratb (Johnes), is often diagnosed this way as large numbers of bacteria are often
present.