期刊
CELLS
卷 11, 期 3, 页码 -出版社
MDPI
DOI: 10.3390/cells11030377
关键词
diethylaminoethyl (DEAE) cellulose; cell separation; cytometry; anti-carp antibody; Sphaerospora; parasite; blood stages; teleost; common carp
类别
资金
- Czech Science Foundation [19-28399X]
- Ministry of Education, Youth and Sport of the Czech Republic [LTAUSA19108, LTAUSA17201]
In this study, a method was developed to isolate myxozoan proliferative stages from fish blood, achieving high purity and survival rates. Additionally, a rapid cytometric assay was developed for quantitative analysis of these stages. This tool will advance functional research on myxozoans and provide important information for the development of therapeutic strategies against myxozoan diseases.
Myxozoans are a diverse group of microscopic cnidarian parasites and some representatives are associated with important diseases in fish, in both marine and freshwater aquaculture systems. Research on myxozoans has been largely hampered by the inability to isolate myxozoan parasites from their host tissues. In this study, we developed and optimized a method to isolate the myxozoan proliferative stages of different size and cellularity from fish blood, using DEAE-cellulose ion exchange chromatography. We optimized several parameters and obtained 99-100% parasite purity, as well as high survival and infectivity. Using polyclonal pan-carp blood cell-specific antibodies, we further developed a rapid cytometric assay for quantification of the proliferative stages, not only in highly concentrated DEAE-C isolates but also in dilute conditions in full blood. Early developmental stages of myxozoans are key to parasite proliferation, establishment, and pathology in their hosts. The isolation of these stages not only opens new possibilities for in vivo and in vitro studies, but also for obtaining purified DNA and protein extracts for downstream analyses. Hence, we provide a long-desired tool that will advance the functional research into the mechanisms of host exploitation and immune stimulation/evasion in this group, which could contribute greatly to the development of therapeutic strategies against myxozoans.
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