Protocol for cryopreservation of the turbot parasite Philasterides dicentrarchi (Ciliophora, Scuticociliatia)

• Published in: Cryobiology
• Main Authors: Folgueira López, Iria, Felipe González, Ana Paula de, Sueiro Benavides, Rosa Ana, Lamas Fernández, Jesús, Leiro Vidal, José Manuel
• Other Authors: Universidade de Santiago de Compostela. Departamento de Bioloxía Funcional, Universidade de Santiago de Compostela. Departamento de Microbioloxía e Parasitoloxía, Universidade de Santiago de Compostela. Instituto de Acuicultura, Universidade de Santiago de Compostela. Instituto de Investigación e Análises Alimentarias
• Format: Article in Journal/Newspaper
• Language: English
• Published: Elsevier
• Subjects: Philasterides dicentrarchi; Miamiensis avidus; Freezing; Cryoprotectants; Cryopreservation; Optimization; Lamas; Leiro; Turbot
• Online Access: http://hdl.handle.net/10347/18172; https://doi.org/10.1016/j.cryobiol.2017.11.010

This is the accepted manuscript of the following article: Folgueira, I., de Felipe, A.P., Sueiro, R.A., Lamas, J. & Leiro, J. (2018). Protocol for cryopreservation of the turbot parasite Philasterides dicentrarchi (Ciliophora, Scuticociliatia). Cryobiology, 80, 77-83. doi:10.1016/j.cryobiol.2017.11.010 Philasterides dicentrarchi is a free-living marine ciliate that can become an endoparasite that causes a severe disease called scuticociliatosis in cultured fish. Long-term maintenance of this scuticociliate in the laboratory is currently only possible by subculture, with periodic passage in fish to maintain the virulence of the isolates. In this study, we developed and optimized a cryopreservation protocol similar to that used for the long-term storage of scuticociliates of the genus Miamiensis. The cryogenic medium comprised ATCC medium 1651 and a combination of 11% dimethylsulfoxide and 5% glycerol. We have verified that the most important factor ensuring the efficiency of the cryopreservation procedure is the growth phase of the culture, and that ciliates should be cryopreserved at the stationary phase (around the sixth day of culture). The cryopreservation protocol described here can be used for all strains of P. dicentrarchi as well as commercial strains of Miamiensis and enables the virulence of the strains to be maintained. Finally, this cryopreservation protocol has been shown to be more effective than others routinely applied to scuticociliates, yielding a higher survival rate with a lower initial concentration of ciliates. The results obtained indicate that the cropreservation protocol enables the long-term storage of scuticociliate parasites while maintaining the virulence of the isolates. The protocol is therefore suitable for use in vaccine production and related studies This work was financially supported by the European Union’s Horizon 2020 research and innovation programme under grant agreement No. 634429 (PARAFISHCONTROL), by the Ministerio de Economía y Competitividad (Spain) under grant ...