Use of Androcoll-ETM to Separate Frozen-Thawed Llama Sperm From Seminal Plasma and Diluent
| Autor: | Fernanda Gabriela Fumuso, Maria Ignacia Carretero, Crissthel Yverlin Guillén Palomino, Mariana Lucía Bertuzzi, Maria Victoria Bariani, S. Giuliano, Nicolás Velásquez González |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
endocrine system
Semen ANDROCOLL Semen analysis cryopreservation Electroejaculation law.invention 03 medical and health sciences 0302 clinical medicine law medicine biology.domesticated_animal SEMEN colloid Acrosome llama Sperm motility Original Research Androcoll COLLOID lcsh:Veterinary medicine diluent 030219 obstetrics & reproductive medicine Chromatography medicine.diagnostic_test biology General Veterinary SEMINAL PLASMA Chemistry urogenital system Extender 0402 animal and dairy science purl.org/becyt/ford/4.4 [https] semen 04 agricultural and veterinary sciences CRYOPRESERVATION LLAMA 040201 dairy & animal science Sperm Lama glama DILUENT seminal plasma lcsh:SF600-1100 Veterinary Science purl.org/pe-repo/ocde/ford#4.03.01 [https] purl.org/becyt/ford/4 [https] |
| Zdroj: | CONICET Digital (CONICET) Consejo Nacional de Investigaciones Científicas y Técnicas instacron:CONICET Instituto Nacional de Innovación Agraria INIA-Institucional instacron:INIA Repositorio Institucional-INIA Frontiers in Veterinary Science Frontiers in Veterinary Science, Vol 7 (2021) |
| DOI: | 10.3389/fvets.2020.594926/full |
| Popis: | 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. Fil: Guillén Palomino, Crissthel Yverlin. Instituto Nacional de Innovación Agraria; Perú Fil: Fumuso, Fernanda Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; Argentina Fil: Bertuzzi, Mariana Lucía. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; Argentina Fil: Giuliano, Susana María. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; Argentina Fil: Velásquez González, Nicolás. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; Argentina Fil: Bariani, Maria Victoria. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Carretero, Maria Ignacia. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina |
| Databáze: | OpenAIRE |
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