Artículo
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, Maria Victoria
; Carretero, Maria Ignacia
Fecha de publicación:
01/2021
Editorial:
Frontiers Media
Revista:
Frontiers in Veterinary Science
ISSN:
2297-1769
Idioma:
Inglés
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
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.
Palabras clave:
ANDROCOLL
,
COLLOID
,
CRYOPRESERVATION
,
DILUENT
,
LLAMA
,
SEMEN
,
SEMINAL PLASMA
Archivos asociados
Licencia
Identificadores
Colecciones
Articulos(OCA PQUE. CENTENARIO)
Articulos de OFICINA DE COORDINACION ADMINISTRATIVA PQUE. CENTENARIO
Articulos de OFICINA DE COORDINACION ADMINISTRATIVA PQUE. CENTENARIO
Citación
Guillén Palomino, Crissthel Yverlin; Fumuso, Fernanda Gabriela; Bertuzzi, Mariana Lucía; Giuliano, Susana María; Velásquez González, Nicolás; et al.; Use of Androcoll-ETM to Separate Frozen-Thawed Llama Sperm From Seminal Plasma and Diluent; Frontiers Media; Frontiers in Veterinary Science; 7; 1-2021; 1-7
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