Use of Androcoll-ETM to Separate Frozen-Thawed Llama Sperm From Seminal Plasma and Diluent
It is not easy to separate frozen-thawed South American camelid sperm from seminal plasma (SP) and diluents to be used for in vitro embryo production. The objective of this study was to evaluate Androcoll-E™ (AE) efficiency to separate llama sperm from SP and freezing extender in frozen-thawed semen...
| Autores principales: | , , , , , |
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| Formato: | info:eu-repo/semantics/article |
| Lenguaje: | Español |
| Publicado: |
Universidad Austral de Chile
2021
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| Materias: | |
| Acceso en línea: | https://hdl.handle.net/20.500.12955/1452 https://doi.org/10.3389/fvets.2020.594926 |
| _version_ | 1855028744594391040 |
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| author | Guillén Palomino, Crissthel Yverlin Fumuso, Fernanda Gabriela Bertuzzi, Mariana Lucía Giuliano, Susana María Velásquez González, Nicolás Bariani, María Victoria |
| author_browse | Bariani, María Victoria Bertuzzi, Mariana Lucía Fumuso, Fernanda Gabriela Giuliano, Susana María Guillén Palomino, Crissthel Yverlin Velásquez González, Nicolás |
| author_facet | Guillén Palomino, Crissthel Yverlin Fumuso, Fernanda Gabriela Bertuzzi, Mariana Lucía Giuliano, Susana María Velásquez González, Nicolás Bariani, María Victoria |
| author_sort | Guillén Palomino, Crissthel Yverlin |
| collection | Repositorio INIA |
| description | It is not easy to separate frozen-thawed South American camelid sperm from seminal plasma (SP) and diluents to be used for in vitro embryo production. The objective of this study was to evaluate Androcoll-E™ (AE) efficiency to separate llama sperm from SP and freezing extender in frozen-thawed semen. A total of 22 ejaculates from five Lama glama males were collected using electroejaculation. After performing semen analysis (sperm motility, concentration, viability, membrane function, and acrosome integrity), samples were cryopreserved with a diluent containing lactose, ethylenediaminetetraacetic acid (EDTA), egg yolk, and 7% dimethylformamide. After thawing, samples were divided in aliquots, one of which was used as a control and the others processed by AE. Experiment 1 (12 ejaculates): 100 μl of frozen-thawed semen was placed on top of 1,000 μl AE column and centrifuged at 800 g for 10 min. Experiment 2 (10 ejaculates): two samples of 100 μl of frozen-thawed semen were placed on two columns of 500 μl AE each, and both were centrifuged at 800 g for 10 and 20 min, respectively. Pellets were resuspended in Tyrode's albumin lactate pyruvate (TALP) medium, and sperm parameters were evaluated. A significant decrease in all sperm parameters was observed in thawed samples compared to raw semen. AE allowed the separation of frozen-thawed sperm from SP and freezing extender independently from the height of the column used and time of centrifugation assayed. Although no significant differences were found between AE columns, higher sperm recovery was observed with 500 μl of AE coupled with 20 min of centrifugation. Despite the significant decrease observed in sperm motility in AE samples, no changes in sperm viability, membrane function, and acrosome integrity were observed when comparing control thawed semen with the sperm recovered after AE (p > 0.05). The use of AE columns, either 500 or 1,000 μl, allows the separation of frozen-thawed llama sperm from SP and freezing extender, preserving the viability, membrane function, and acrosome integrity. Of the protocols studied, 800 g centrifugation during 20 min using a 500 μl column of AE would be the method of choice to process frozen-thawed llama semen destined for reproductive biotechnologies. |
| format | info:eu-repo/semantics/article |
| id | INIA1452 |
| institution | Institucional Nacional de Innovación Agraria |
| language | Español |
| publishDate | 2021 |
| publishDateRange | 2021 |
| publishDateSort | 2021 |
| publisher | Universidad Austral de Chile |
| publisherStr | Universidad Austral de Chile |
| record_format | dspace |
| spelling | INIA14522022-11-21T20:16:52Z Use of Androcoll-ETM to Separate Frozen-Thawed Llama Sperm From Seminal Plasma and Diluent Guillén Palomino, Crissthel Yverlin Fumuso, Fernanda Gabriela Bertuzzi, Mariana Lucía Giuliano, Susana María Velásquez González, Nicolás Bariani, María Victoria Androcoll Colloid Cryopreservation Diluent Llama Semen Seminal plasma https://purl.org/pe-repo/ocde/ford#4.03.01 It is not easy to separate frozen-thawed South American camelid sperm from seminal plasma (SP) and diluents to be used for in vitro embryo production. The objective of this study was to evaluate Androcoll-E™ (AE) efficiency to separate llama sperm from SP and freezing extender in frozen-thawed semen. A total of 22 ejaculates from five Lama glama males were collected using electroejaculation. After performing semen analysis (sperm motility, concentration, viability, membrane function, and acrosome integrity), samples were cryopreserved with a diluent containing lactose, ethylenediaminetetraacetic acid (EDTA), egg yolk, and 7% dimethylformamide. After thawing, samples were divided in aliquots, one of which was used as a control and the others processed by AE. Experiment 1 (12 ejaculates): 100 μl of frozen-thawed semen was placed on top of 1,000 μl AE column and centrifuged at 800 g for 10 min. Experiment 2 (10 ejaculates): two samples of 100 μl of frozen-thawed semen were placed on two columns of 500 μl AE each, and both were centrifuged at 800 g for 10 and 20 min, respectively. Pellets were resuspended in Tyrode's albumin lactate pyruvate (TALP) medium, and sperm parameters were evaluated. A significant decrease in all sperm parameters was observed in thawed samples compared to raw semen. AE allowed the separation of frozen-thawed sperm from SP and freezing extender independently from the height of the column used and time of centrifugation assayed. Although no significant differences were found between AE columns, higher sperm recovery was observed with 500 μl of AE coupled with 20 min of centrifugation. Despite the significant decrease observed in sperm motility in AE samples, no changes in sperm viability, membrane function, and acrosome integrity were observed when comparing control thawed semen with the sperm recovered after AE (p > 0.05). The use of AE columns, either 500 or 1,000 μl, allows the separation of frozen-thawed llama sperm from SP and freezing extender, preserving the viability, membrane function, and acrosome integrity. Of the protocols studied, 800 g centrifugation during 20 min using a 500 μl column of AE would be the method of choice to process frozen-thawed llama semen destined for reproductive biotechnologies. INTRODUCTION. MATERIALS AND METHODS. RESULTS. DISCUSSION. CONCLUSIONS. REFERENCES. 2021-08-31T20:16:34Z 2021-08-31T20:16:34Z 2021-01-21 info:eu-repo/semantics/article Guillén Palomino CY, Fumuso FG, Bertuzzi ML, Giuliano SM, Velásquez González N, Bariani MV and Carretero MI (2021) Use of Androcoll-ETM to Separate Frozen-Thawed Llama Sperm From Seminal Plasma and Diluent. Front. Vet. Sci. 7:594926. doi: 10.3389/fvets.2020.594926 https://hdl.handle.net/20.500.12955/1452 Frontiers in Veterinary Science https://doi.org/10.3389/fvets.2020.594926 spa Frontiers in Veterinary Science Volume 7: 594926 https://doi.org/10.3389/fvets.2020.594926 info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf application/pdf Perú Universidad Austral de Chile Suiza Instituto Nacional de Innovación Agraria Repositorio Institucional - INIA |
| spellingShingle | Androcoll Colloid Cryopreservation Diluent Llama Semen Seminal plasma https://purl.org/pe-repo/ocde/ford#4.03.01 Guillén Palomino, Crissthel Yverlin Fumuso, Fernanda Gabriela Bertuzzi, Mariana Lucía Giuliano, Susana María Velásquez González, Nicolás Bariani, María Victoria Use of Androcoll-ETM to Separate Frozen-Thawed Llama Sperm From Seminal Plasma and Diluent |
| title | Use of Androcoll-ETM to Separate Frozen-Thawed Llama Sperm From Seminal Plasma and Diluent |
| title_full | Use of Androcoll-ETM to Separate Frozen-Thawed Llama Sperm From Seminal Plasma and Diluent |
| title_fullStr | Use of Androcoll-ETM to Separate Frozen-Thawed Llama Sperm From Seminal Plasma and Diluent |
| title_full_unstemmed | Use of Androcoll-ETM to Separate Frozen-Thawed Llama Sperm From Seminal Plasma and Diluent |
| title_short | Use of Androcoll-ETM to Separate Frozen-Thawed Llama Sperm From Seminal Plasma and Diluent |
| title_sort | use of androcoll etm to separate frozen thawed llama sperm from seminal plasma and diluent |
| topic | Androcoll Colloid Cryopreservation Diluent Llama Semen Seminal plasma https://purl.org/pe-repo/ocde/ford#4.03.01 |
| url | https://hdl.handle.net/20.500.12955/1452 https://doi.org/10.3389/fvets.2020.594926 |
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