A cryopreservation procedure for the rumen protozoon Entodinium caudatum: estimation of its viability by fluorescence microscopy

de la Fuente Oliver, Gabriel; Cebrián, J. A.; Fondevila, Manuel

doi:https://doi.org/10.1111/j.1472-765X.2003.01464.x

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Issue date

2004

Author

de la Fuente Oliver, Gabriel

Cebrián, J. A.

Fondevila, Manuel

Suggested citation

de la Fuente Oliver, Gabriel; Cebrián, J. A.; Fondevila, Manuel; . (2004) . A cryopreservation procedure for the rumen protozoon Entodinium caudatum: estimation of its viability by fluorescence microscopy. Letters in Applied Microbiology, 2004, vol. 38, núm. 2, p. 164-168. https://doi.org/10.1111/j.1472-765X.2003.01464.x.

Abstract

Aims: To study the viability of a culture of the rumen protozoon Entodinium caudatum after a cryopreservation procedure by a fluorescence microscopy staining method. Methods and Results: Fluorescence method is based on the different colour of cells depending on their membrane integrity. When the temperature effect was studied either by fluorescence or motility, the techniques were correlated (r = 0·727) and their slopes and intercepts were not different (P > 0·05). However, motility showed a higher variation coefficient (0·40 vs 0·12). There were no differences between cooling rates at cryopreservation (1 and 4°C min−1) at 38, 15 or 5°C, nor after thawing. Conclusions: Fluorescence staining is more accurate than motility for assessing protozoal viability. Viability after thawing was 0·50, and the number of viable cells per 250 μl straw was 320 and 420 for 1 and 4°C min−1. Significance and Impact of the Study: This cryopreservation procedure seems to ensure culture recovery for E. caudatum.

URI

http://hdl.handle.net/10459.1/49222

DOI

https://doi.org/10.1111/j.1472-765X.2003.01464.x

Is part of

Letters in Applied Microbiology, 2004, vol. 38, núm. 2, p. 164-168