Effects of Thawing Procedure on Postthawed In Vitro Viability and In Vivo Fertility of Red Deer Epididymal Spermatozoa Cryopreserved at −196°C

Soler, Ana Josefa; García, Andrés J.; Fernández-Santos, M.R.; Esteso, M.C.; Garde, José Julián

doi:10.1002/j.1939-4640.2003.tb02737.x

Cited by 59 publications

(61 citation statements)

References 52 publications

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“…Para su evaluación y enfriamiento el tubo con los espermatozoides diluidos se traspasaron a otro recipiente de 200 ml de capacidad y fueron colocados dentro una refrigeradora a 5°C por espacio de 3 h. Para la congelación se adiciono 0.5 mL de volumen de dilutor que contenía el 14% de glicerol y cada dosis se ajustaron a 20 X 10 de espermatozoides motiles en 6 pajillas de 0.25 mL, se sellaron con polivinilo de cloruro, se mantuvieron a 5°C por espacio de 45 min para su equilibrio. Para la congelación se utilizó el protocolo descrito por Soler et al (2003). La descongelación de las pajillas se realizó a una temperatura de 37°C por espacio de 30 s.…”

Section: Procesamiento De Los Espermatozoidesunclassified

“…De la misma forma comparando con material espermático colectado de los epidídimos de animales post mortem, como es del ovino, vacuno y ciervo ibérico, los investigadores reportan motilidades a la descongelación del 35 al 70% (Kaabi et al 2003;Ehling et al 2006;Herold et al 2004;Martins et al 2007;Garde y col. 1998;Soler et al 2003;Soler et al 2005;Fernandez-Santos et al 2006), estas motilidades totales son superiores, comparados con las motilidades del reproductor 1 de 24.68% y para el reproductor 2 de 27.31%, encontrados en el presente estudio.…”

Section: Discusión Prueba De Fertilidad In-vitro De Los Espermatozoidunclassified

“…Comparando los resultados obtenidos a la prueba hipo-osmótica a la descongelación de los espermatozoides en el presente trabajo de 32.18% para el reproductor 2 y 20.06% para el reproductor 1, resulta ser inferior frente a los resultados obtenidos en espermatozoides del epidídimo de ciervos cazados y posterior congelación en el mismo tipo de dilutor (Triladyl), los autores a la descongelación y a la prueba hipo-osmótica reportaron porcentajes del 44.1 al 62.7% (Garde y col. 1998;Soler et al 2003), esta superioridad se debe a que estos autores señalan que las muestras iníciales de motilidad estuvieron por encima del 90%, coincidiendo que en todo el protocolo de congelación/descongelación adecuado el 50% de espermatozoides mueren producto del stress de la congelación/descongelación. Mientras que en el presente estudio la proporción de espermatozoides que murieron durante el presente protocolo de congelación fue del 45%, porcentaje a c e p t a d o c u a n d o s e e v a l ú a m u e s t r a s d e espermatozoides congelados en laboratorio.…”

Section: Discusión Prueba De Fertilidad In-vitro De Los Espermatozoidunclassified

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Viabilidad in vitro e in vivo de los espermatozoides congelados/descongelados del conducto deferente de Alpacas (vicugna pacos)

Durand¹,

Aragón²,

Guerra³

2016

Rev. Investig. Altoandin. - RIA -

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KEY WORDS:Viabilidad in vitro e in vivo de los espermatozoides congelados/descongelados del conducto deferente de alpacas (Vicugna pacos)Viability in vitro and invivo of sperm frozen/thawed from alpaca (Vicugna pacos) vas deferens El objetivo del presente estudio fue evaluar la viabilidad in-vitro e in-vivo de los espermatozoides procedente de los conductos deferentes de alpacas. Se utilizaron 2 reproductores como donadores de espermatozoides con desviación del conducto deferente. Los espermatozoides colectados se sometieron a la congelación y descongelación con el dilutor Triladyl®. Durante el procesamiento de los espermatozoides se evaluaron la motilidad y la prueba hipo-osmotica. En la inseminación se evaluó la proporción de gestaciones producidas. Los resultados fueron los siguientes: La motilidad de los espermatozoides para los reproductores 1 y 2 fueron: a los 37°C 65.16 y 63.37% sin diferencia (P>0.05), al enfriamiento (5°C) 52.63 y 48.31% similares (P>0.05) ya la descongelación 24.51 y 33.18% mostraron diferencia (P<0.05). A la prueba hipo-osmótica fueron; a los 37°C 51.55 y 52.98% similares (P>0.05), Al enfriamiento (5°C) 46.67 y 55.93% mostraron diferencia (P<0.05) y a la descongelación 20.06 y 32.18% con diferencia (P<0.05). Las gestaciones fueron: Con espermatozoides frescos diluidos 36.36% (4/11), con espermatozoides descongelados 25.00% (5/20) y con monta 54.54% (6/11), siendo similares (P>0.05). En conclusión los espermatozoides procedentes de los conductos deferentes soportan el congelamiento y a la inseminación artificial producen gestaciones.The aim of this study was to evaluate the in-vitro and in-vivo viability of sperm from the vas deferens alpacas. 2 males as sperm donors were used to offset the vas deferens. The collected sperm cells were subjected to freezing and thawing with dilutor Triladyl®. During processing of sperm motility and hypo-osmotic test they were evaluated. Insemination in the proportion of pregnancies produced was evaluated. The results were as follows: The sperm motility of males 1 and 2 were: at 37 ° C 65.16 and 63.37% with no difference (P> 0.05), cooling (5 ° C) 52.63 and 48.31% similar (P > 0.05) and thawing 24.51 and 33.18% showed no difference (P <0.05). The hypo-osmotic test were; at 37 ° C 51.55 and 52.98 similar percentage (P> 0.05), and cooling 55.93% 46.67 showed no difference (P <0.05) and thawing 20.06 and 32.18% with difference (P <0.05). Pregnancies were diluted with fresh sperm 36.36% (4/11), with defrosted sperm 25.00% (5/20) and 54.54% mounts (6/11), being similar (P> 0.05). In conclusion sperm from the vas deferens withstand freezing and insemination are able to produce pregnancies.

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Section: Procesamiento De Los Espermatozoidesunclassified

Section: Discusión Prueba De Fertilidad In-vitro De Los Espermatozoidunclassified

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Viabilidad in vitro e in vivo de los espermatozoides congelados/descongelados del conducto deferente de Alpacas (vicugna pacos)

Durand¹,

Aragón²,

Guerra³

2016

Rev. Investig. Altoandin. - RIA -

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“…To allow the insemination of a large number of females with spermatozoa from the same male, sperm samples were cryopreserved in liquid nitrogen after collection. A detailed description of methods used for sperm freezing and artificial insemination can be found elsewhere [40]. After sample thawing and before insemination, subsamples were taken for evaluation of several sperm parameters.…”

Section: Artificial Insemination Trialsmentioning

confidence: 99%

Male Fertility in Natural Populations of Red Deer Is Determined by Sperm Velocity and the Proportion of Normal Spermatozoa1

Malo

Garde

García

et al. 2005

Biology of Reproduction

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222

209

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Male reproductive success is determined by the ability of males to gain sexual access to females and by their ability to fertilize ova. Among polygynous mammals, males differ markedly in their reproductive success, and a great deal of effort has been made to understand how selective forces have shaped traits that enhance male competitiveness both before and after copulation (i.e., sperm competition). However, the possibility that males also may differ in their fertility has been ignored under the assumption that male infertility is rare in natural populations because selection against it is likely to be strong. In the present study, we examined which semen traits correlate with male fertility in natural populations of Iberian red deer (Cervus elaphus hispanicus). We found no trade-offs between semen traits. Our analyses revealed strong associations between sperm production and sperm swimming velocity, sperm motility and proportion of morphologically normal spermatozoa, and sperm viability and acrosome integrity. These last two variables had the lowest coefficients of variation, suggesting that these traits have stabilized at high values and are unlikely to be related to fitness. In a fertility trial, our results show a large degree of variation in male fertility, and differences in fertility were determined mainly by sperm swimming velocity and by the proportion of morphologically normal sperm. We conclude that male fertility varies substantially in natural populations of Iberian red deer and that, when sperm numbers are equal, it is determined mainly by sperm swimming velocity and sperm morphology.

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“…Amostras oriundas de epidídimos refrigerados a 5°C durante 24 h podem ser aproveitadas para uma posterior congelação em diferentes espécies (Marks et al 1994, Blash et al 2002, Soler et al 2003.…”

Section: Discussionunclassified

Refrigeração do epidídimo canino a 4ºC e recuperação dos espermatozoides epididimários utilizando ACP-106c

et al. 2013

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. 2013. Objetivou-se avaliar a qualidade dos espermatozoides recuperados da cauda do epidídimo após a refrigeração do complexo testículo-epidídimo (CTE) de cães usando o diluidor ACP-106c. Foram utilizados 60 cães machos adultos, com peso de 10-20 kg.Após a eutanásia, removeu-se o CTE que foi imerso em solução fisiológica 0,9% e transportado em caixa térmica ao laboratório a 30°C. Para a refrigeração e recuperação dos espermatozoides epididimários, os 60 pares do CTE foram divididos em 4 grupos, de acordo com o tempo de refrigeração do CTE e posterior recuperação espermática: G0h, G6h, G12h e G18h, em que cada par do CTE permaneceu por zero, seis, doze ou dezoito horas a 4°C, respectivamente. A recuperação dos espermatozoides da cauda do epidídimo foi realizada pela técnica de flutuação utilizando-se o diluidor ACP-106c ou Tris. Para cada epidídimo foi adicionado 1,0 mL de um dos dois diluidores, pré-aquecidos a 37°C por 5 minutos. Em seguida foram centrifugados a 800g/5 minutos para remoção dos resíduos celulares. Avaliou-se The study aimed to assess the quality of sperm recovered from the epididymal cauda after cooling the complex testis-epididymis (CTE) of dogs using ACP-106c extender. Sixty adult male dogs, weighing 10-20 kg were used. After euthanasia, CTE was removed and it was immersed in 0.9% saline and transported to the laboratory in cooler at 30°C. For cooling and recovery of epididymal spermatozoa, the 60 pairs of CTE were divided into four groups, according to the refrigeration time of the CTE and subsequent sperm recovery: G0h, G6h, G12h and G18h, wherein each pair of CTE remained zero, six, 12 or 18 hours at 4°C respectively. The recovery of sperm epididymal tail was conducted by flotation technique using ACP-106c or Tris extender. For each epididymis, it was added 1.0 mL of either extenders, preheated to 37°C for 5 minutes. They were then centrifuged at 800g/5 minutes to remove the cell debris. Morphology, functionality and total and progressive motility, and parameters obtained by CASA were evaluated. Data were analyzed by ANOVA followed by Turkey test (P <0.05). In all parameters assessed, there was no difference between the extenders used (P>0.05). The values of total motility groups G0h, G6H, G12H, and G18h for ACP-106c were 84.4±7.7, 81.6±11.6, 88.3±6.5 and 69.5±16.9 respectively, and for Tris 85.2±8.7, 77.4±14.3, 79.0±17.8 and 65.4±17.9 respectively. A decrease in sperm quality was observed after 18 hours of cooling in both extenders. Thus it can be concluded that the ACP-106c may be used to recover the epididymal spermatozoa chilled and may be viable for up to 12h cooling.INDEX TERMS: Refrigeration, epididymis, dogs.

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Effects of Thawing Procedure on Postthawed In Vitro Viability and In Vivo Fertility of Red Deer Epididymal Spermatozoa Cryopreserved at −196°C

Cited by 59 publications

References 52 publications

Viabilidad in vitro e in vivo de los espermatozoides congelados/descongelados del conducto deferente de Alpacas (vicugna pacos)

Viabilidad in vitro e in vivo de los espermatozoides congelados/descongelados del conducto deferente de Alpacas (vicugna pacos)

Male Fertility in Natural Populations of Red Deer Is Determined by Sperm Velocity and the Proportion of Normal Spermatozoa1

Refrigeração do epidídimo canino a 4ºC e recuperação dos espermatozoides epididimários utilizando ACP-106c

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