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Effective freezing rate for semen cryopreservation in endangered Mediterranean brown trout (Salmo trutta macrostigma) inhabiting the Biferno river (South Italy)

Published online by Cambridge University Press:  28 December 2015

Nicolaia Iaffaldano*
Affiliation:
Department of Agricultural, Environmental and Food Sciences, University of Molise, via De Sanctis snc, 86100 Campobasso, Italy. Department of Agricultural, Environmental and Food Sciences, University of Molise, 86100Italy.
Michele Di Iorio
Affiliation:
Department of Agricultural, Environmental and Food Sciences, University of Molise, 86100Italy.
Angelo Manchisi
Affiliation:
Department of Agricultural, Environmental and Food Sciences, University of Molise, 86100Italy.
Stefano Esposito
Affiliation:
Mediterranean Trout Research Group–Centro di ricerche ‘I Giardini dell’Acqua’, 42037 Collagna (RE), Italy.
Pier Paolo Gibertoni*
Affiliation:
Department of Agricultural, Environmental and Food Sciences, University of Molise, via De Sanctis snc, 86100 Campobasso, Italy. Mediterranean Trout Research Group–Centro di ricerche ‘I Giardini dell’Acqua’, 42037 Collagna (RE), Italy.
*
All correspondence to: Nicolaia Iaffaldano, Department of Agricultural, Environmental and Food Sciences, University of Molise, via De Sanctis snc, 86100 Campobasso, Italy. Tel. +39 0874 404697. Fax +39 0874 404855. E-mail: nicolaia@unimol.it
Pier Paolo Gibertoni, Mediterranean Trout Research Group–Centro di ricerche ‘I Giardini dell’Acqua’ – 42037 Collagna (RE), Italy. E-mail: gibertoni@igiardinidellacqua.com

Summary

This study was designed to determine: (i) the in vitro effects of different freezing rates on post-thaw semen quality of Mediterranean brown trout (Salmo trutta macrostigma) from the Biferno river; and (ii) the in vivo fertilization and hatching percentage of freezing rate giving rise to the best post-thaw semen quality.

Pooled semen samples were diluted 1:3 (v:v) in a freezing extender composed of 300 mM glucose, 10% egg yolk and 10% dimethyl sulfoxide (DMSO). The extended semen was packaged in 0.25 ml plastic straws and frozen at different heights above the liquid nitrogen surface (1, 5 or 10 cm) for 10 min to give three different freezing rates. Semen samples were thawed at 30°C for 10 s. The variables assessed after thawing were sperm motility, duration of motility and viability.

Our results clearly indicate a significant effect of freezing rate on post-thaw semen quality. Semen frozen 5 cm above the liquid nitrogen surface showed the best quality after freezing/thawing. Based on these in vitro data, 2 groups of 200 eggs were fertilized with fresh semen or semen frozen 5 cm above the liquid nitrogen surface. Fertilization and hatching rates recorded for eggs fertilized with frozen semen were significantly lower (25.4% and 22.5%, respectively) than the ones obtained using fresh semen (87.8% and 75.5%, respectively). An effective freezing protocol will allow for the creation of a sperm cryobank to recover the original population of Mediterranean brown trout in the Biferno river.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2015 

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