Cryopreservation of semen in the dog: use of ultra-freezers of −152 °C as a viable alternative to liquid nitrogen

Alamo, D; Batista, M.; González, F.; Rodríguez, Nathalie; Cruz, Guadalupe Santa; Cabrera, Fernando; Gracia, A.

doi:10.1016/j.theriogenology.2004.03.016

Cited by 38 publications

(40 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Further studies with larger numbers are needed to determine the usefulness of this technique for freezing equine sperm. Alamo et al (2005) reported on the use of ultra-freezers of −152 • C as a viable alternative to liquid nitrogen for freezing dog spermatozoa. Sperm motility recorded with the use of ultra-low temperature freezers at −152 • C was comparable to that obtained with liquid nitrogen.…”

Section: Advances In Freezing Semenmentioning

confidence: 99%

Integration of future biotechnologies into the equine industry

Squires¹

2005

Animal Reproduction Science

View full text Add to dashboard Cite

There has and will continue to be reproductive techniques available that have a positive impact upon the equine breeding industry. This review focuses on semen technologies that have been developed or are in the process of being developed. The use of fluorescent dyes and flow cytometry has provided the researcher and clinician with powerful tools to evaluate several sperm attributes. These procedures have been utilized to evaluate sperm viability, acrosome status, mitochondrial status, DNA integrity and stages of capacitation. Flow cytometry allows several sperm attributes to be evaluated on thousands of spermatozoa in a matter of seconds. Development of procedures for insemination of mares with relatively small numbers of spermatozoa has the potential to change how stallions and their semen are managed. This review discusses the use of insemination of fresh, frozen and sex-sorted spermatozoa in relatively small numbers compared with conventional insemination technologies. The recent acceptance of frozen-thawed semen by many of the major breed registries has stimulated an increase in research on frozen semen. Many of the studies have focused on identifying damage during the freezing and thawing process. Numerous studies also have been conducted to modify freezing extenders so that the sperm are protected during the freezing and thawing process. The production of in vitro-produced embryos is extremely limited in the horse due to the failure of in vitro fertilization. However, intracytoplasmic sperm injection (ICSI) has been used for the production of foals from stallions that have less than typical sperm numbers or from stallions that have died and a limited quantity of frozen semen is available. This technique has been used by several laboratories to produce embryos in vitro. The breeder and veterinarian now have access to techniques that allow assessment of semen quality, improvement of procedures for freezing and thawing and insemination of mares with fewer numbers of spermatozoa. It is likely that the next decade will also produce tremendous advances in semen technologies that can be utilized in the horse industry.

show abstract

Section: Advances In Freezing Semenmentioning

confidence: 99%

Integration of future biotechnologies into the equine industry

Squires¹

2005

Animal Reproduction Science

View full text Add to dashboard Cite

show abstract

“…El efecto beneficioso de la utilización de metodologías alternativas al N 2 L para criopreservar células espermáticas ha sido reportado previamente (Salamon, 1971;Medrano et al, 2002;Alamo et al;Batista et al), sin embargo, el uso de ultracongeladores no programables para congelar/almacenar semen ha sido de uso limitado (Alamo et al; Batista et al). No existen antecedentes previos de la utilización del ultracongelador de -80ºC como técnica para conservar y almacenar espermatozoides caninos, siendo este el primer estudio que evalúa esta posibilidad, analizando por citometría de flujo la función espermática.…”

Section: Discussionunclassified

“…En caninos, la congelación y conservación de espermatozoides por periodos prolongados de tiempo se realiza en nitrógeno líquido (N 2 L) (Alamo et al, 2005). Además, han sido utilizadas técnicas alternativas para el procedimiento de congelación como el hielo seco y ultra-congeladores programables para conservar muestras de semen, sin embargo el almacenamiento es en N 2 L (Olar et al, 1989;Hay et al, 1997;Thomas et al, 1993).…”

Section: Introductionunclassified

Criopreservación de Espermatozoides Caninos a - 80C

2013

View full text Add to dashboard Cite

RESUMEN. La congelación y almacenamiento en nitrógeno líquido (N 2 L) es la técnica más utilizada para preservar el semen de canino y de otras especies domésticas durante largos períodos de tiempo. Este estudio fue diseñado para validar el uso del ultracongelador de -80ºC para congelar y almacenar semen canino en pajuelas. Tres protocolos fueron ensayados (n=10): Control: semen congelado y almacenado en N 2 L; Exp. 1: semen congelado y almacenado -80ºC, y Exp.2: semen congelado a -80ºC y almacenado en N 2 L. Postdescongelación se evaluó por citometría de flujo la viabilidad e integridad de membrana plasmática (SYBR-14/PI), potencial de membrana mitocondrial (∆Ψm; JC-1), integridad de la membrana del acrosoma (PSA/FITC-PI), translocación de fosfatidilserina (Annexin-V-FITC/PI) y fragmentación del ADN (TUNEL). No se observaron diferencias significativas (P<0,05) entre los grupos Control, Exp.1 y Exp.2 respecto a: integridad de membrana plasmática intacta (42,2 ± 5,3; 35,57 ± 10,3 y 40,76 ± 12,1; respectivamente), ∆Ψm normal (54,7 ± 20,6; 44,4 ± 15,8 y 43,4 ± 15,3 respectivamente), membrana acrosomal intacta (42,9 ± 11,1; 53,2 ± 15,8 y 48,7 ± 20,1 respectivamente) y ADN fragmentado (0,87 ± 0,41; 0,75 ± 0,16; 0,79 ± 0,27 respectivamente). Sin embargo, el promedio de la motilidad progresiva postdescongelación de los grupos E1 y E2 (37,0 ± 4,4%; 36,8 ± 4,1% respectivamente) demostraron diferencias (P<0,05) respecto al grupo Control (55,9 ± 8,7%). Los resultados obtenidos en el presente estudio demostraron que la utilización del ultracongelador de -80ºC para congelar y almacenar espermatozoides caninos tiene un uso potencial en medicina veterinaria como alternativa a la utilización del N 2 L. PALABRAS CLAVE: Canino; Espermatozoide; Congelación; Ultracongelador de -80ºC. INTRODUCCIÓNEn caninos, la congelación y conservación de espermatozoides por periodos prolongados de tiempo se realiza en nitrógeno líquido (N 2 L) (Alamo et al., 2005). Además, han sido utilizadas técnicas alternativas para el procedimiento de congelación como el hielo seco y ultra-congeladores programables para conservar muestras de semen, sin embargo el almacenamiento es en N 2 L (Olar et al., 1989;Hay et al., 1997;Thomas et al., 1993).Ha sido determinada la función espermática en ultracongeladores de -152ºC utilizando parámetros como la motilidad, reacción del acrosoma y morfoanomalías (Ála-mo et al.;Batista et al., 2006). Sin embargo, no existen antecedentes de la utilización del ultracongelador de -80ºC, para congelar semen canino en pajuelas convencionales por periodos cortos como alternativa a la técnica convencional de congelación en N 2 L. En este estudio se evalúa el efecto de la temperatura de congelación y almacenamiento a -80ºC, en comparación al método convencional de congelación y almacenamiento en N 2 L, sobre parámetros esenciales de la función del espermatozoide de canino, tales como motilidad progresiva, funcionalidad de membranas (plasmática, acrosomal y mitocondrial) y fragmentación del ADN. MATERIAL Y MÉTODOAnimales. Se utilizaron...

show abstract

“…In addition to the traditional method for freezing semen in straws, freezers operating with liquid nitrogen are also used (2,3,4,11). However, with the changes in life sciences, liquid nitrogen-based systems have faced with some problems, the most important two being the contamination risk and temperature change up to 90 °C (4,6,7,8,10).…”

mentioning

confidence: 99%

“…However, with the changes in life sciences, liquid nitrogen-based systems have faced with some problems, the most important two being the contamination risk and temperature change up to 90 °C (4,6,7,8,10). Therefore, there is a need to investigate feasible alternatives for semen cryopreservation which can prevent the complications of traditional methods (2). However, there are also other studies confirming that another viable alternative may be the use of ultra-low freezers at -152 °C, not too far from liquid nitrogen temperature at -196 °C (2,3,4,11).…”

mentioning

confidence: 99%

Effect of alternative cryopreservation procedures on bull semen

Korkmaz;DAŞKIN¹

2012

Ankara Üniversitesi Veteriner Fakültesi Dergisi

View full text Add to dashboard Cite

Summary:The aim of this study was to assess the use of -152°C ultra freezer for freezing and storing bull semen as an alternative to freezing over the liquid nitrogen vapor and storing in liquid nitrogen in terms of certain post-thaw spermatological parameters. Two ejaculates, one per each bull, were used without pooling. Four different freezing and storing protocols were tested. At 1 week, 2 months and 6 months after cryopreservation, the frozen semen was thawed and the seminal parameters were determined. As a conclusion, this study has confirmed that the use of -152 °C ultra-low freezer for freezing and storing bull semen for 6 months is a viable alternative to liquid nitrogen.Key words: Bull, cryopreservation, semen, storage, ultra-low freezer. Alternatif Dondurma ve Saklama Yöntemlerinin Boğa Sperması Üzerine EtkisiÖzet: Bu çalışmanın amacı, boğa spermasının -152 °C derin dondurucuda dondurulup saklanmasının sıvı azot buharında dondurma ve sıvı azotta saklamaya çözüm sonu kimi spermatolojik parametreler bakımından bir alternatif olup olmayacağının araştırılmasıdır. Bu çalışmada iki adet boğanın birer ejakulatı miks yapılmadan kullanıldı. Araştırmada dört farklı dondurma ve saklama protokolü denendi. Dondurma işleminden bir hafta, iki ve altı aylık periyotlar sonrasında payetler çözdürüldü ve çözüm sonu bazı spermatolojik özellikler değerlendirildi. Sonuç olarak bu çalışma -152°C derin dondurucunun boğa spermasının dondurulması ve altı ay süreyle saklanmasında sıvı azota bir alternatif olarak kullanılabilirliğini ve boğa spermasının dondurulması ve saklanmasında iyi bir ortam sağladığını doğrulamaktadır. Anahtar sözcükler: Boğa, derin dondurucu, dondurma, saklama, sperma.* Aynı isimli doktora tezinden özetlenmiştir.Bull semen can be frozen in pellets, ampoules or straws and then stored in liquid nitrogen (13). In addition to the traditional method for freezing semen in straws, freezers operating with liquid nitrogen are also used (2,3,4,11). However, with the changes in life sciences, liquid nitrogen-based systems have faced with some problems, the most important two being the contamination risk and temperature change up to 90 °C (4,6,7,8,10). Therefore, there is a need to investigate feasible alternatives for semen cryopreservation which can prevent the complications of traditional methods (2). However, there are also other studies confirming that another viable alternative may be the use of ultra-low freezers at -152 °C, not too far from liquid nitrogen temperature at -196 °C (2,3,4,11). It remains to be established mechanical freezers which can provide air phase at -150°C without the need for liquid nitrogen can prevent cross-contamination of germ-plasm over a long period of storage (6,7). The aim of this experiment was to assess the use of an ultra-low freezer at -152 °C for freezing and storage of bull semen for 6 months. Postthawing seminal parameters were evaluated to see whether an ultra-low freezer at -152 °C could be a feasible alternative to cryopreservation in liquid nitrogen.Two Holstein bulls we...

show abstract

Cryopreservation of semen in the dog: use of ultra-freezers of −152 °C as a viable alternative to liquid nitrogen

Cited by 38 publications

References 21 publications

Integration of future biotechnologies into the equine industry

Integration of future biotechnologies into the equine industry

Criopreservación de Espermatozoides Caninos a - 80C

Effect of alternative cryopreservation procedures on bull semen

Contact Info

Product

Resources

About