Optimization of Sperm Cryopreservation Protocol for Mediterranean Brown Trout: A Comparative Study of Non-Permeating Cryoprotectants and Thawing Rates In Vitro and In Vivo
The aim of our study was to test the effects of different non-permeating cryoprotectants (NP-CPAs), namely low-density lipoproteins (LDLs), sucrose, and egg yolk, and thawing rates on the post-thaw semen quality and fertilizing ability of the native Mediterranean brown trout. Pooled semen samples we...
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MDPI AG
2019-05-01
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| Online Access: | https://www.mdpi.com/2076-2615/9/6/304 |
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| author | Giusy Rusco Michele Di Iorio Pier Paolo Gibertoni Stefano Esposito Maurizio Penserini Alessandra Roncarati Silvia Cerolini Nicolaia Iaffaldano |
| author_facet | Giusy Rusco Michele Di Iorio Pier Paolo Gibertoni Stefano Esposito Maurizio Penserini Alessandra Roncarati Silvia Cerolini Nicolaia Iaffaldano |
| author_sort | Giusy Rusco |
| collection | DOAJ |
| description | The aim of our study was to test the effects of different non-permeating cryoprotectants (NP-CPAs), namely low-density lipoproteins (LDLs), sucrose, and egg yolk, and thawing rates on the post-thaw semen quality and fertilizing ability of the native Mediterranean brown trout. Pooled semen samples were diluted 1:3 (v:v) with 2.5%, 5%, 10%, or 15% LDL; 0.05, 0.1, or 0.3 M sucrose; or 10% egg yolk. At the moment of analysis, semen was thawed at 30 °C/10 s or 10 °C/30 s. The post-thaw semen quality was evaluated, considering motility, the duration of motility, viability, and DNA integrity. Significantly higher values of motility and viability were obtained using egg yolk/10 °C for 30 s, across all treatments. However, LDL and sucrose concentrations affected sperm cryosurvival, showing the highest post-thaw sperm quality at 5% LDL and 0.1 M sucrose. Based on the in vitro data, egg yolk, 5% LDL, and 0.1 M sucrose thawed at 10 °C or 30 °C were tested for the in vivo trial. The highest fertilization and hatching rates were recorded using egg yolk/10 °C (<i>p</i> < 0.05). According to these in vitro and in vivo results, egg yolk emerged as the most suitable NP-CPA and 10 °C/30 s as the best thawing rate for the cryopreservation of this trout sperm, under our experimental conditions. |
| first_indexed | 2024-12-11T19:11:53Z |
| format | Article |
| id | doaj.art-f60b93a946b64ceebba6df014b3f3635 |
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| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-12-11T19:11:53Z |
| publishDate | 2019-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Animals |
| spelling | doaj.art-f60b93a946b64ceebba6df014b3f36352022-12-22T00:53:46ZengMDPI AGAnimals2076-26152019-05-019630410.3390/ani9060304ani9060304Optimization of Sperm Cryopreservation Protocol for Mediterranean Brown Trout: A Comparative Study of Non-Permeating Cryoprotectants and Thawing Rates In Vitro and In VivoGiusy Rusco0Michele Di Iorio1Pier Paolo Gibertoni2Stefano Esposito3Maurizio Penserini4Alessandra Roncarati5Silvia Cerolini6Nicolaia Iaffaldano7Department of Agricultural, Environmental and Food Sciences, University of Molise, 86100 Campobasso CB, ItalyDepartment of Agricultural, Environmental and Food Sciences, University of Molise, 86100 Campobasso CB, ItalyMediterranean Trout Research Group—Centro di ricerche “I Giardini dell’Acqua”, 42037 Collagna RE, ItalyMediterranean Trout Research Group—Centro di ricerche “I Giardini dell’Acqua”, 42037 Collagna RE, ItalyMediterranean Trout Research Group—Centro di ricerche “I Giardini dell’Acqua”, 42037 Collagna RE, ItalySchool of Biosciences and Veterinary Medicine, University of Camerino, 62032 Camerino MC, ItalyDepartment of Veterinary Medicine, University of Milan, 20122 Milano MI, ItalyDepartment of Agricultural, Environmental and Food Sciences, University of Molise, 86100 Campobasso CB, ItalyThe aim of our study was to test the effects of different non-permeating cryoprotectants (NP-CPAs), namely low-density lipoproteins (LDLs), sucrose, and egg yolk, and thawing rates on the post-thaw semen quality and fertilizing ability of the native Mediterranean brown trout. Pooled semen samples were diluted 1:3 (v:v) with 2.5%, 5%, 10%, or 15% LDL; 0.05, 0.1, or 0.3 M sucrose; or 10% egg yolk. At the moment of analysis, semen was thawed at 30 °C/10 s or 10 °C/30 s. The post-thaw semen quality was evaluated, considering motility, the duration of motility, viability, and DNA integrity. Significantly higher values of motility and viability were obtained using egg yolk/10 °C for 30 s, across all treatments. However, LDL and sucrose concentrations affected sperm cryosurvival, showing the highest post-thaw sperm quality at 5% LDL and 0.1 M sucrose. Based on the in vitro data, egg yolk, 5% LDL, and 0.1 M sucrose thawed at 10 °C or 30 °C were tested for the in vivo trial. The highest fertilization and hatching rates were recorded using egg yolk/10 °C (<i>p</i> < 0.05). According to these in vitro and in vivo results, egg yolk emerged as the most suitable NP-CPA and 10 °C/30 s as the best thawing rate for the cryopreservation of this trout sperm, under our experimental conditions.https://www.mdpi.com/2076-2615/9/6/304<i>Salmo cettii</i>sperm freezingegg yolkconservation biologysperm cryobankfertilization rate |
| spellingShingle | Giusy Rusco Michele Di Iorio Pier Paolo Gibertoni Stefano Esposito Maurizio Penserini Alessandra Roncarati Silvia Cerolini Nicolaia Iaffaldano Optimization of Sperm Cryopreservation Protocol for Mediterranean Brown Trout: A Comparative Study of Non-Permeating Cryoprotectants and Thawing Rates In Vitro and In Vivo Animals <i>Salmo cettii</i> sperm freezing egg yolk conservation biology sperm cryobank fertilization rate |
| title | Optimization of Sperm Cryopreservation Protocol for Mediterranean Brown Trout: A Comparative Study of Non-Permeating Cryoprotectants and Thawing Rates In Vitro and In Vivo |
| title_full | Optimization of Sperm Cryopreservation Protocol for Mediterranean Brown Trout: A Comparative Study of Non-Permeating Cryoprotectants and Thawing Rates In Vitro and In Vivo |
| title_fullStr | Optimization of Sperm Cryopreservation Protocol for Mediterranean Brown Trout: A Comparative Study of Non-Permeating Cryoprotectants and Thawing Rates In Vitro and In Vivo |
| title_full_unstemmed | Optimization of Sperm Cryopreservation Protocol for Mediterranean Brown Trout: A Comparative Study of Non-Permeating Cryoprotectants and Thawing Rates In Vitro and In Vivo |
| title_short | Optimization of Sperm Cryopreservation Protocol for Mediterranean Brown Trout: A Comparative Study of Non-Permeating Cryoprotectants and Thawing Rates In Vitro and In Vivo |
| title_sort | optimization of sperm cryopreservation protocol for mediterranean brown trout a comparative study of non permeating cryoprotectants and thawing rates in vitro and in vivo |
| topic | <i>Salmo cettii</i> sperm freezing egg yolk conservation biology sperm cryobank fertilization rate |
| url | https://www.mdpi.com/2076-2615/9/6/304 |
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