Tetracapsuloides Bryosalmonae (Myxozoa: Malacosporea) Portal of Entry into the Fish Host

The portal of entry and the penetration process of the myxozoan parasite Tetracapsuloides bryosalmonae are still poorly understood. In the present study, spores of T. bryosalmonae derived from the bryozoan host (malacospores) were activated chemically and mechanically to investigate their reaction after attachment to the fish host in vitro. Amoeboid movement of both sporoplasms was shown for the first time. The morphology of malacospores was assessed using scanning electron microscopy (SEM). Openings of the polar capsules and released polar filaments were visible. One sporoplasm was observed leaving the spore shell. Laboratory exposure experiments of juvenile rainbow trout Oncorhynchus mykiss to spores of T. bryosalmonae were also conducted. Single fish were incubated with 1000 to 2000 spores in 100 ml of water for 5 to 60 min. Immunohistochemically stained sections of skin and gills were examined using light microscopy, as well as ultra thin sections using transmission electron microscopy (TEM), to investigate attachment and early penetration. Whole fish and excised gills of fish exposed to a spore suspension were fixed and prepared for scanning electron microscopy. Attached and penetrating stages were found only on or in the gills, and not in the skin. Due to the low overall number of spores, only a few spores were found adjacent to the gill epithelium in TEM. No parasite stages were found on the samples used for SEM. These results indicate that the gills are the preferred entry loci for the amoeboid sporoplasms of T. bryosalmonae into the fish host.