Abstract

To facilitate detailed investigation of inflammatory and degenerative changes of harbor porpoise (Phocoena phocoena) ears immunohistochemical and special staining techniques were implemented. Therefore, two monoclonal markers as well as three polyclonal antibodies of different species specificities directed against different antigens were tested for immunohistochemical cross-reactivity on formalin-fixed, paraffin-embedded middle and inner ear sections of a harbor porpoise with Stenurus minor infection. Additionally, von Kossa, Luxol Fast Blue, Bielschofsky and Cresyl violet special stainings for the detection of degenerative changes of nerve fibers and neurons rather than?? chromatolysis, respectively, were performed. All investigated antibodies showed specific immunoreactivities with leukocytes in harbor porpoise ears. Lymphocytes were labeled by a polyclonal human-specific anti-CD3 antibody and a monoclonal CD79α-marker. Cells of the histiocytic lineage were recognized by a human polyclonal anti-lysozyme-antibody and the monoclonal MAC 387-antibody. The MHC class II-antigen was labeled by a bovine-specific monoclonal antibody. Mineralization was detected in tissue sections by von Kossa silver staining. Myelin and myelinated axons, respectively, were stained by Luxol Fast Blue staining. Furthermore, axons were visualized by Bielschofsky special staining and Nissl substance was detected in the cytoplasm of neurons by Cresyl violet staining. This study showed the applicability of leukocyte markers for the characterization of inflammatory responses in formalin-fixed, paraffin embedded tissue sections of the ear of the harbor porpoise by immunohistochemistry. Additionally performed special staining techniques enable the detection of neuronal structures and degenerative changes, respectively, in harbor porpoise's ears.

Acknowledgements

This study is funded by the Federal Ministry of Food, Agriculture and Consumer Protection, through the Federal Agency for Agriculture and Food, "Potential impact of pingers (acoustic deterrent devices) in EU fisheries on harbor porpoises" (06HS010).

The authors also wish to thank Bettina Buck for excellent technical support.