Abstract
Inclusion body disease (IBD) is a worldwide disease in captive boa constrictors (boa constrictor) and occasionally in other snakes of the families Boidae and Pythonidae. The exact causative agent(s) and pathogenesis are not yet fully understood. Currently, diagnosis of IBD is based on the light microscopic identification of eosinophilic intracytoplasmic inclusion bodies in hematoxylin and eosin stained tissues or blood smears. An antigenically unique 68 KDa protein was identified within the IBD inclusion bodies, called IBD protein. A validated immuno-based ante-mortem diagnostic test is needed for screening snakes that are at risk of having IBD. In this study, despite difficulties in solubilizing semi-purified inclusion bodies, utilizing hybridoma technology a mouse anti-IBD protein monoclonal antibody (MAB) was produced. The antigenic specificity of the antibody was confirmed and validated by western blots, enzyme-linked immunosorbent assay, immuno-transmission electron microscopy, and immunohistochemical staining. Paraffin embedded tissues of IBD positive and negative boa constrictors (n=94) collected from 1990 to 2011 were tested with immunohistochemical staining. In boa constrictors, the anti-IBDP MAB had a sensitivity of 83% and specificity of 100% in detecting IBD. The antibody also cross-reacted with IBD inclusion bodies in carpet pythons (Morelia spilota) and a ball python (python regius). This validated antibody can serve as a tool for the development of ante-mortem immunodiagnostic tests for IBD.
Highlights
Inclusion Body Disease (IBD) is a unique and insidious worldwide disease commonly seen in captive members of snakes in the families Boidae and Pythonidae [1]
Paraffin embedded tissues of 99 snakes were collected from the repositories of Anatomic Pathology Service and Zoological Medicine Infectious Disease Testing Laboratory, College of Veterinary Medicine, University of Florida (UF)(Gainesville, FL, USA), and additional blocks were kindly provided by Northwest ZooPath (Monroe, WA, USA) and Zoo/Exotic Pathology Services (West Sacramento, CA, USA)
The IBD- liver did not result in any solid pellet after the incubation with sarkosyl, whereas a tightly bound pellet was obtained from all three IBD+ samples (Figure 1)
Summary
Inclusion Body Disease (IBD) is a unique and insidious worldwide disease commonly seen in captive members of snakes in the families Boidae and Pythonidae [1]. The disease was most commonly reported in boa constrictors (Boa constrictor) and occasionally in other related snakes [1,2,3,4,5,6,7,8,9,10,11,12,13,14]. Inclusion body disease is characterized by observation of eosinophilic intracytoplasmic inclusion bodies in hematoxylin and eosin (H&E) stained histological slides or blood smears, which is the current gold standard for diagnosing IBD [1,12]. Inclusion bodies seen in IBD sometimes can be difficult to distinguish from other cellular proteinaceus material or cellular granules that may accumulate in the cytoplasm of affected cells [1]. A validated molecular diagnostic test that is more sensitive and specific is needed
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