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/G Lester laughton Page 1 3/15/2011

Granulomas. The Meat Works lesion.


Background. All reactor animals are thoroughly checked Post mortem for lesions. This
is a term for any mass, growth, or abnormality. The lymph nodes are especially checked
both externally, and by slicing, as these are the “collecting “ organs associated with the
immune system. The immune system is essentially carried to all tissues via the blood
stream. The white blood cells are responsible for this.
Lesions found are cut into two pieces; one smaller one is fixed in 10% formalin. As this
will only penetrate a small way through the tissue, samples should not be more than
10mm thick. The other sample is as big as possible to place in the pottle.
The FIXED sample is used for Histology. Here the sample is flooded with liquid paraffin
wax, which sets solid. From this solid mass, very thin slices, one cell thick are cut. These
are then glued on a microscope slide and stained with appropriate stains depending on
what is being looked for.
The FRESH sample is held refrigerated until the nature of the changes in the lesion are
determined by Histology..looking at the slide under high powered microscope.

Inflammation, or The inflammatory response, is triggered at any site where an insult


or injury occurs.
This starts with a dilation of local blood vessels in response to prostaglandin and
histamine release locally.
There is also increased permeability of these small blood vessels. This allows further
fluid and blood contents to accumulate, causing Heat, Pain and Swelling. These are the
three classical signs of inflammation
The first White blood cells to arrive at an inflammatory site are the Neutrophils. These
are able to pass out of blood vessels and accumulate at the site of injury or infection.
They are often referred to as Pus cells.
At this time there is often fibrin deposited in the area, which then forms into strands, and
so forms a Fibrous capsule around the site.
These early stages may be referred to as the Acute stage of an infection or injury. This
could be a Pus filled abscess.
In the longer term, or more Chronic stages, referred to as Granulomas, there are a variety
of other white blood cells, which may concentrate at the site, which ones depends on the
type of infection or injury.
Lymphocytes will be more prevalent if a chronic bacterial infection is occurring (The
Mycobacteria group, Actinobacteria)
Eosinophils are prominent in any Allergic response. This we see in Parasitic reactions.
Macrophages, Giant cells, (Langahans Giant cells on Multi-nucleated Giant cells, for the
Mycobacteria) are seen in the still later stages of chronic infections. With these cells the
nucleus has divided several times but not the cell cytoplasm, and so there are several
nucleuses within one “Giant” cell, which also will contain Mycobacteria. (AFOs see
later)
At this later stage of a chronic infection a very thick capsule will often form in order to
try and “wall off” the infection. Throughout the lesion the more fluid pus becomes dryer,
and cheesy in consistency. This is referred to as Caseous pus. It often has some areas that
become calcified, which gives the gritty sensation when cutting such a lesion. These
changes can be seen when the tissue is stained with special stains.
/G Lester laughton Page 2 3/15/2011

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.

Actinobacillosis is also chronic granulomatous forming bacteria infection, commonly


referred to as Woody Tongue. These are not acid fast bacteria, and can usually be seen as
“Club like Colonies” within the macrophages. Usually this is a clear indication of Actino
infection.

Parasites; eosinophils dominate the granuloma reaction, but other cells are also present.

Tumors are characterized by showing many dividing cells, as there is “uncontrolled


growth” in the lesion.

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.