Effects of glycerol and sugar mixing temperature on the morphologic and functional integrity of cryopreserved ram sperm

Autores
Pelufo, Valeria; Lopez Armengol, Maria Fernanda; Malcotti, Valeria; Venturino, Andres; Aisen, Eduardo Gabriel
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Sperm deep freezing procedures for ram semen have considerable variations regarding the steps being employed for cooling, freezing, and addition of cryoprotectants. In this work, we evaluated the effects of the addition of glycerol and/or the disaccharides sucrose and trehalose to hypertonic diluents either before or after cooling from 30°C to 5°C in Merino Australian ram semen cryopreservation. Using optical and transmission electron microscopy techniques, we assessed that glycerol was beneficial to the cooling process independently of its addition at 30°C or 5°C in terms of sperm membrane integrity in different regions of the plasma membrane (acrosomal region, 14.5% higher integrity; postacrosomal region, 8.0% higher integrity [P<0.01]; hypoosmotic swelling test [HOST], 10.8% higher integrity [P<0.001]). Disaccharides were necessary for a better cryopreservation in liquid nitrogen, and the best procedure was their addition after cooling at 5°C (12% higher sperm motility [P<0.001]; 8% higher acrosome integrity, [P<0.05]; 9.5% higher plasma membrane integrity assessed by HOST [P<0.001]). Trehalose showed a greater preservation cryoprotectant capacity than sucrose, as indicated by sperm motility after thawing (8.1% greater [P<0.01]) and by the integrity of the intermediate piece (20% greater [P<0.05]). From these results, we conclude that the best procedure for ram semen cryopreservation in hypertonic disaccharide-containing diluents is the addition of glycerol and trehalose after the cooling process, at 5°C.
Fil: Pelufo, Valeria. Universidad Nacional del Comahue. Facultad de Ciencias Agrarias. Instituto de Biotecnología Agropecuaria del Comahue; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Lopez Armengol, Maria Fernanda. Universidad Nacional del Comahue. Facultad de Ciencias Agrarias. Instituto de Biotecnología Agropecuaria del Comahue; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Malcotti, Valeria. Gobierno de la Provincia del Neuquén. Ministerio de Desarrollo Territorial; Argentina
Fil: Venturino, Andres. Universidad Nacional del Comahue. Facultad de Ciencias Agrarias. Instituto de Biotecnología Agropecuaria del Comahue; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Aisen, Eduardo Gabriel. Universidad Nacional del Comahue. Facultad de Ciencias Agrarias. Instituto de Biotecnología Agropecuaria del Comahue; Argentina
Materia
Cooling Process
Frozen-Thawed Ram Spermatozoa
Plasma Membrane Domains
Trehalose
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/37853
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/37853
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Effects of glycerol and sugar mixing temperature on the morphologic and functional integrity of cryopreserved ram spermPelufo, ValeriaLopez Armengol, Maria FernandaMalcotti, ValeriaVenturino, AndresAisen, Eduardo GabrielCooling ProcessFrozen-Thawed Ram SpermatozoaPlasma Membrane DomainsTrehalosehttps://purl.org/becyt/ford/4.4https://purl.org/becyt/ford/4Sperm deep freezing procedures for ram semen have considerable variations regarding the steps being employed for cooling, freezing, and addition of cryoprotectants. In this work, we evaluated the effects of the addition of glycerol and/or the disaccharides sucrose and trehalose to hypertonic diluents either before or after cooling from 30°C to 5°C in Merino Australian ram semen cryopreservation. Using optical and transmission electron microscopy techniques, we assessed that glycerol was beneficial to the cooling process independently of its addition at 30°C or 5°C in terms of sperm membrane integrity in different regions of the plasma membrane (acrosomal region, 14.5% higher integrity; postacrosomal region, 8.0% higher integrity [P<0.01]; hypoosmotic swelling test [HOST], 10.8% higher integrity [P<0.001]). Disaccharides were necessary for a better cryopreservation in liquid nitrogen, and the best procedure was their addition after cooling at 5°C (12% higher sperm motility [P<0.001]; 8% higher acrosome integrity, [P<0.05]; 9.5% higher plasma membrane integrity assessed by HOST [P<0.001]). Trehalose showed a greater preservation cryoprotectant capacity than sucrose, as indicated by sperm motility after thawing (8.1% greater [P<0.01]) and by the integrity of the intermediate piece (20% greater [P<0.05]). From these results, we conclude that the best procedure for ram semen cryopreservation in hypertonic disaccharide-containing diluents is the addition of glycerol and trehalose after the cooling process, at 5°C.Fil: Pelufo, Valeria. Universidad Nacional del Comahue. Facultad de Ciencias Agrarias. Instituto de Biotecnología Agropecuaria del Comahue; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Lopez Armengol, Maria Fernanda. Universidad Nacional del Comahue. Facultad de Ciencias Agrarias. Instituto de Biotecnología Agropecuaria del Comahue; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Malcotti, Valeria. Gobierno de la Provincia del Neuquén. Ministerio de Desarrollo Territorial; ArgentinaFil: Venturino, Andres. Universidad Nacional del Comahue. Facultad de Ciencias Agrarias. Instituto de Biotecnología Agropecuaria del Comahue; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Aisen, Eduardo Gabriel. Universidad Nacional del Comahue. Facultad de Ciencias Agrarias. Instituto de Biotecnología Agropecuaria del Comahue; ArgentinaElsevier Science Inc2015-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/37853Pelufo, Valeria; Lopez Armengol, Maria Fernanda; Malcotti, Valeria; Venturino, Andres; Aisen, Eduardo Gabriel; Effects of glycerol and sugar mixing temperature on the morphologic and functional integrity of cryopreserved ram sperm; Elsevier Science Inc; Theriogenology; 83; 1; 1-2015; 144-1510093-691XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.theriogenology.2014.09.007info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0093691X14004968info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:01:51Zoai:ri.conicet.gov.ar:11336/37853instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-22 11:01:51.749CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Effects of glycerol and sugar mixing temperature on the morphologic and functional integrity of cryopreserved ram sperm
title Effects of glycerol and sugar mixing temperature on the morphologic and functional integrity of cryopreserved ram sperm
spellingShingle Effects of glycerol and sugar mixing temperature on the morphologic and functional integrity of cryopreserved ram sperm
Pelufo, Valeria
Cooling Process
Frozen-Thawed Ram Spermatozoa
Plasma Membrane Domains
Trehalose
title_short Effects of glycerol and sugar mixing temperature on the morphologic and functional integrity of cryopreserved ram sperm
title_full Effects of glycerol and sugar mixing temperature on the morphologic and functional integrity of cryopreserved ram sperm
title_fullStr Effects of glycerol and sugar mixing temperature on the morphologic and functional integrity of cryopreserved ram sperm
title_full_unstemmed Effects of glycerol and sugar mixing temperature on the morphologic and functional integrity of cryopreserved ram sperm
title_sort Effects of glycerol and sugar mixing temperature on the morphologic and functional integrity of cryopreserved ram sperm
dc.creator.none.fl_str_mv Pelufo, Valeria
Lopez Armengol, Maria Fernanda
Malcotti, Valeria
Venturino, Andres
Aisen, Eduardo Gabriel
author Pelufo, Valeria
author_facet Pelufo, Valeria
Lopez Armengol, Maria Fernanda
Malcotti, Valeria
Venturino, Andres
Aisen, Eduardo Gabriel
author_role author
author2 Lopez Armengol, Maria Fernanda
Malcotti, Valeria
Venturino, Andres
Aisen, Eduardo Gabriel
author2_role author
author
author
author
dc.subject.none.fl_str_mv Cooling Process
Frozen-Thawed Ram Spermatozoa
Plasma Membrane Domains
Trehalose
topic Cooling Process
Frozen-Thawed Ram Spermatozoa
Plasma Membrane Domains
Trehalose
purl_subject.fl_str_mv https://purl.org/becyt/ford/4.4
https://purl.org/becyt/ford/4
dc.description.none.fl_txt_mv Sperm deep freezing procedures for ram semen have considerable variations regarding the steps being employed for cooling, freezing, and addition of cryoprotectants. In this work, we evaluated the effects of the addition of glycerol and/or the disaccharides sucrose and trehalose to hypertonic diluents either before or after cooling from 30°C to 5°C in Merino Australian ram semen cryopreservation. Using optical and transmission electron microscopy techniques, we assessed that glycerol was beneficial to the cooling process independently of its addition at 30°C or 5°C in terms of sperm membrane integrity in different regions of the plasma membrane (acrosomal region, 14.5% higher integrity; postacrosomal region, 8.0% higher integrity [P<0.01]; hypoosmotic swelling test [HOST], 10.8% higher integrity [P<0.001]). Disaccharides were necessary for a better cryopreservation in liquid nitrogen, and the best procedure was their addition after cooling at 5°C (12% higher sperm motility [P<0.001]; 8% higher acrosome integrity, [P<0.05]; 9.5% higher plasma membrane integrity assessed by HOST [P<0.001]). Trehalose showed a greater preservation cryoprotectant capacity than sucrose, as indicated by sperm motility after thawing (8.1% greater [P<0.01]) and by the integrity of the intermediate piece (20% greater [P<0.05]). From these results, we conclude that the best procedure for ram semen cryopreservation in hypertonic disaccharide-containing diluents is the addition of glycerol and trehalose after the cooling process, at 5°C.
Fil: Pelufo, Valeria. Universidad Nacional del Comahue. Facultad de Ciencias Agrarias. Instituto de Biotecnología Agropecuaria del Comahue; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Lopez Armengol, Maria Fernanda. Universidad Nacional del Comahue. Facultad de Ciencias Agrarias. Instituto de Biotecnología Agropecuaria del Comahue; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Malcotti, Valeria. Gobierno de la Provincia del Neuquén. Ministerio de Desarrollo Territorial; Argentina
Fil: Venturino, Andres. Universidad Nacional del Comahue. Facultad de Ciencias Agrarias. Instituto de Biotecnología Agropecuaria del Comahue; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Aisen, Eduardo Gabriel. Universidad Nacional del Comahue. Facultad de Ciencias Agrarias. Instituto de Biotecnología Agropecuaria del Comahue; Argentina
description Sperm deep freezing procedures for ram semen have considerable variations regarding the steps being employed for cooling, freezing, and addition of cryoprotectants. In this work, we evaluated the effects of the addition of glycerol and/or the disaccharides sucrose and trehalose to hypertonic diluents either before or after cooling from 30°C to 5°C in Merino Australian ram semen cryopreservation. Using optical and transmission electron microscopy techniques, we assessed that glycerol was beneficial to the cooling process independently of its addition at 30°C or 5°C in terms of sperm membrane integrity in different regions of the plasma membrane (acrosomal region, 14.5% higher integrity; postacrosomal region, 8.0% higher integrity [P<0.01]; hypoosmotic swelling test [HOST], 10.8% higher integrity [P<0.001]). Disaccharides were necessary for a better cryopreservation in liquid nitrogen, and the best procedure was their addition after cooling at 5°C (12% higher sperm motility [P<0.001]; 8% higher acrosome integrity, [P<0.05]; 9.5% higher plasma membrane integrity assessed by HOST [P<0.001]). Trehalose showed a greater preservation cryoprotectant capacity than sucrose, as indicated by sperm motility after thawing (8.1% greater [P<0.01]) and by the integrity of the intermediate piece (20% greater [P<0.05]). From these results, we conclude that the best procedure for ram semen cryopreservation in hypertonic disaccharide-containing diluents is the addition of glycerol and trehalose after the cooling process, at 5°C.
publishDate 2015
dc.date.none.fl_str_mv 2015-01
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/37853
Pelufo, Valeria; Lopez Armengol, Maria Fernanda; Malcotti, Valeria; Venturino, Andres; Aisen, Eduardo Gabriel; Effects of glycerol and sugar mixing temperature on the morphologic and functional integrity of cryopreserved ram sperm; Elsevier Science Inc; Theriogenology; 83; 1; 1-2015; 144-151
0093-691X
CONICET Digital
CONICET
url http://hdl.handle.net/11336/37853
identifier_str_mv Pelufo, Valeria; Lopez Armengol, Maria Fernanda; Malcotti, Valeria; Venturino, Andres; Aisen, Eduardo Gabriel; Effects of glycerol and sugar mixing temperature on the morphologic and functional integrity of cryopreserved ram sperm; Elsevier Science Inc; Theriogenology; 83; 1; 1-2015; 144-151
0093-691X
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.theriogenology.2014.09.007
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0093691X14004968
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier Science Inc
publisher.none.fl_str_mv Elsevier Science Inc
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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