Immunohistochemical Study of Rabies Virus Within the Central Nervous System of Domestic and Wildlife Species

Abstract

Immunohistochemistry using a commercial polyclonal antibody for lyssavirus was applied to 39 archival cases of rabies. Paraffin blocks from 13 different species were available, including 3 dogs, 4 cats, 1 pig, 6 cattle, 4 horses, 1 llama, 7 skunks (Mephitis mephitis), 7 raccoons (Procyon lotor), 1 bat (Myotis species), 1 white-tailed deer (Odocoileus virginianus), 1 bobcat (Lynx rufus), 2 gray foxes (Urocyon cinereoargenteus), and 1 red fox (Vulpes vulpes). All cases had previously been diagnosed as rabies using histopathology and/or fluorescent antibody testing. The immunohistochemistry technique successfully detected lyssavirus antigen in all cases. In species for which 3 or more samples were available, distributional trends were seen in 4 main brain regions: brainstem, cerebellum, hippocampus, and cerebrum. The best site for rabies virus detection in dogs and cats was the hippocampus. For cattle, viral antigen was most prominent in the brainstem, followed by the cerebellum. In horses, the cervical spinal cord and adjacent brainstem were the optimal sites for detecting rabies virus antigen. In raccoons and skunks, positive labeling was widely dispersed, so selection might be less important for these wildlife reservoir species. Immunohistochemistry should prove useful in enhancing the accuracy of rabies diagnosis through informed selection of brain sampling sites when composite sampling is not feasible. This immunohistochemical technique could provide reliable virus detection in formalin-fixed tissues in any potentially infected species.

Calbindin D28k distribution in neurons and reactive gliosis in cerebellar cortex of natural Rabies virus–infected cattle

“…37 These findings agree with previous reports in cattle and other herbivores, 4,25,27,43 which are unlike the immunoreactivity in the cerebellum of RABV-infected horses. 5,37 The IHC protocol used here confirms its utility as a safe alternative to FAT, by preserving the tissue architecture and allowing histologic evaluation to reach a diagnosis. 25,29,33,34,37 GFAP immunoreactivity showed a mild astroglial reaction in CNS tissue, thus confirming our results from HE histopathology and a previous report that used another type of GFAP antibody in RABV-infected cattle, 28 along with multifocal microglial proliferation.…”

“…29,33,37 In agreement with these reports, the pAbs accurately detected RABV in FFPE samples from the CNS of cattle, probably with more sensitivity than monoclonal primary antibodies, because pAbs would potentially detect a wider range of RABV strains, and could be useful to differentiate other non-rabid inclusion bodies in bovine CNS in cases where rabies was suspected. 37 In the cerebellum, Purkinje neurons were the best neuronal type to study Negri bodies, and also to evaluate RABV immunoreactivity. Furthermore, in the cerebellar cortex, other immunopositive large interneurons were identified in the granular layer, probably Golgi and/or Lugaro cells, in a pattern similar to that which was previously reported.…”

“…5,37 The IHC protocol used here confirms its utility as a safe alternative to FAT, by preserving the tissue architecture and allowing histologic evaluation to reach a diagnosis. 25,29,33,34,37 GFAP immunoreactivity showed a mild astroglial reaction in CNS tissue, thus confirming our results from HE histopathology and a previous report that used another type of GFAP antibody in RABV-infected cattle, 28 along with multifocal microglial proliferation. In the cerebellar cortex, patches of reactive Bergmann glia were evident occasionally in the cerebellar molecular layer with GFAP, accompanied by extensive multifocal proliferation of microglia organized as Babès nodules, which agrees with other reports.…”

Resumos Do Xvi Enapave

“…São descritas meningoencefalite mononuclear perivascular, múltiplos focos de gliose (Notici et al 2009;Pessoa et al 2011), além de áreas de malácia com infiltrado de macrófagos (Notici et al 2009) e neuroniofagia (Pessoa et al 2011). A imunofluorescência direta é o teste padrão ouro para o diagnóstico de raiva em animais e humanos por ser um método rápido e sensível (WHO, 2004) Todavia a imuno-histoquímica pode ser utilizada para demonstração do antígeno rábico em tecidos que foram submetidos a fixação em formalina (Stein et al, 2010). Este trabalho tem por objetivo relatar a ocorrência de dois casos de raiva em suínos, diagnosticados através de imuno-histoquimica.…”

Pneumomediastinum and subcutaneous emphysema in a rabid bull

“…No lesions were observed in the other organs. Rabies virus antigen was detected in the Purkinje neurons, spinal cord and obex ( Figure 1D) using a goat anti-rabies polyclonal reagent (Chemicon #5199) according to the technique described by STEIN et al (2010).…”