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Diseases of Aquatic Organisms

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DAO 42:215-219 (2000)  -  doi:10.3354/dao042215

Prolonged in vitro cultivation of Ichthyophthirius multifiliis using an EPC cell line as substrate

C. V. Nielsen, K. Buchmann*

Department of Veterinary Microbiology, Section of Fish Diseases, Royal Veterinary and Agricultural University, Stigbøjlen 4, 1870 Frederiksberg C, Denmark
*Corresponding author. E-mail:

ABSTRACT: The ciliate Ichthyophthirius multifiliis, which normally requires a fish host to develop from the theront stage to the trophont stage, was cultivated in vitro for part of its life cycle. Experiments were conducted using a laboratory strain of the parasite originally isolated from rainbow trout Oncorhynchus mykiss in a Danish trout farm. Theronts escaping from tomontocysts were kept in water, cell culture media (E-MEM or L-15), or cultures of EPC (Epithelioma Papulosum Cyprini) cells in plastic tissue culture dishes (Nunc multidish plates). In addition, a 2-compartment system, with water separated from tissue culture media by a monolayer of EPC cells on an Anopore Tissue Culture Insert (mimicking the fish epidermis) was tested as an experimental habitat for the parasite. Theronts transformed into trophonts in all treatments except in water alone. However, development was accelerated in wells containing EPC cells, and survival and growth of trophonts were significantly increased compared to water or tissue culture media alone. Further, the 2-compartment system allowed superior performance of the parasites (attachment of parasites to cells and growth from 36 to 46 µm). In all experiments it was found that the presence of host factors (mucus and serum) stimulated parasite development.

KEY WORDS: In vitro culture · Parasite · Ciliate · Ichthyophthirius multifiliis · EPC cells · E-MEM · L-15

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