The present study deals with Berlin strains of Histomonas meleagridis, the specimens of which were cultivated in Dwyer's medium. The light and electron microscopic examination revealed that the cultivated trophozoite stages (reaching about 10 mum in size) appeared more or less spherical, although their surface (covered by a single membrane) showed amoeba-like waves. All stages were uni-nucleated and reproduced by binary fission with an extranuclear spindle apparatus. Some trophozoites appeared ovoid and possessed a single flagellum with a typical microtubular 9 x 2 + 2 arrangement. Furthermore, the latter were characterized by an inner row of typical microtubules (remnant of an axostyle) and a Golgi apparatus (both adjacent to the nucleus), multivesicular structures, hydrogenosomes, and many food vacuoles containing either starch grains or bacteria. Their cytoplasm was densely filled with glycogen granules and ribosomes. Similar stages were also documented in the caeca and cloaca of chicken when being inoculated (via cloaca) with such culture stages. In addition to these typical trophozoites, the cultures contained a low number of 10-mum-sized spherical cyst-like stages with a surrounding amorphous layer. The cytoplasm of some of these cyst-like stages-when studied by electron microscopy-appeared with two membranes or had formed an amorphic, cyst-wall-like layer at their surface, apparently corresponding to their light microscopical appearance. Such stages might be involved in transmission from one host to another and probably have been missed before in microscopical examinations of infected poultry.
Berlin strains of Histomonas meleagridis were subcultivated to produce cyst-like stages. These strains were studied for their ITS 1 and 18S rRNA properties and compared with sequences in data banks of other H. meleagridis strains, Dientamoeba fragilis, and some species of the genus Trichomonas and relatives. The Berlin isolates that had previously been shown to be able to develop cyst-like structures (Munsch et al. 2008) represent a significant cluster among the published data of other Histomonas meleagridis isolates and thus the formation of cysts might be a common feature that would open further possibilities of transmission.
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