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RE: Questions about IBD testing info

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Posted by: raptor1 at Tue Jul 10 03:46:53 2007   [ Email Message ] [ Show All Posts by raptor1 ]  
   

Hi Nick As I understand - the only sure fire way to find if a boa does have Ibd is on a dead animal (testing all major organs)
But... If aquiring a new animal it may be wise to have a blood test done ( i think you may be able to have a liver biopsy done too on a live animal with just a local anasethic) Anyhow with the bood test ( H&E staining) if any of the results come back with posetive results for incusion bodies - you treat this boa as suspect. You can obviously watch for the classic symptoms while in quarentine, also have the boas tested again 3 months later. Intrestingly I have read (been trying to trawl the forums to find this info so I could post it - no luck) bottom line is 50% of boas tested in a study for Ibd tested posetive!
Now that is scary! Here is more info and links

http://sacs.vetmed.ufl.edu/Old Files/wildlife/IBDINFO.html


http://www.anapsid.org/ibd.html





Diagnosis: Currently there is no serologic assay available for determining exposure. If one of the retroviruses we have isolated is found to be the causative agent, we will be able to start developing a serological test. Boa constrictor immunoglobulin has been purified and a mice have been inoculated to produce a polyclonal against this immunoglobulin. At the University of Florida College of Veterinary Medicine, we perform complete blood counts on suspect snakes. Infected snakes commonly have white blood cells counts >30,000/ul. Intracytoplasmic inclusions are occasionally seen in peripheral lymphocytes (Figure 10). We also take esophageal, gastric, and liver bopsies.

To make a more rapid diagnosis, cytologic smears can be made from biopsies and stained using a modified hematoxylin and eosin (H&E) technique:

1. Fix 1 minute in 10% neutral buffered formalin
2. Stain 3 minutes in Harris hematoxylin
3. One dip into acid alcohol
4. Briefly wash in running water
5. Dip into 0.5%ammonia to blue the nuclei
6. Wash in running water
7. Counterstain in eosin for 40 seconds
8. Dehydrate in a series of alcohols: 95%-100%-100%
9. Place in xylene and mount as with a paraffin embedded section

I have found this staining far superior than using Wright-Giemsa staining. With Wright Giemsa staining the inclusions stain basophilic and may not be readily recognizable. With H&E staining, inclusions stain exactly the same as in tissues embedded in paraffin, sectioned and stained with H&E. It must be remembered that absence of inclusions in the biopsies does not necessarily mean the snake is free of IBD. In some cases inclusions may only be seen in the central nervous system, and in those cases may be few in number. The diagnosis is only as good as the portion sampled and biopsies routinely represent a relatively small portion of the entire tissue.


   

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