Effect of semen dilution to low-sperm number per dose on motility and functionality of cryopreserved bovine spermatozoa using low-density lipoproteins (LDL) extender: Comparison to Triladyl® and Bioxcell®

Vera-Munoz, O.; Amirat-Briand, L.; Díaz, Thais; Vásquez, Luís; Schmidt, E. T.; Desherces, S.; Anton, Marc; Bencharif, D.; Tainturier, D.

doi:10.1016/j.theriogenology.2008.10.010

Cited by 42 publications

(26 citation statements)

References 22 publications

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“…The higher volume of seminal plasma would seem to be beneficial for cryopreservation, 11 but not for refrigeration at 4 uC for more than 3…”

Section: Discussionmentioning

confidence: 99%

“…Many other studies have confirmed that the LDL fraction has cryoprotective action. [8][9][10][11] Egg yolk can therefore be replaced with fractions of LDLs in semen extenders for freezing. An advantage of an LDL extender over a standard egg yolk extender is that it has a less complex chemical composition and does not interfere with microscopic observations.…”

Section: Introductionmentioning

confidence: 99%

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Effect of low-density lipoproteins, spermatozoa concentration and glycerol on functional and motility parameters of bull spermatozoa during storage at 4 °C

Vera-Munoz¹,

Amirat-Briand²,

Bencharif³

et al. 2010

Asian J Androl

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An extender has been developed with low-density lipoproteins (LDLs) that eliminates the microbial risks associated with the use of whole egg yolk. The objective of this study was to assess the effects of substituting egg yolk with LDLs for use as an extender in sperm preservation at 4 6C, as well as on spermatozoa motility, plasma membrane and acrosome integrity, at two different concentrations (80310 6 and 240310 6 sperm per ml) for 8 days and to evaluate glycerol toxicity in both extenders. A total of 12 ejaculates were collected from three bulls. Spermatozoa motility was examined using computer-assisted semen analysis. Plasma membrane integrity was determined using the hypo-osmotic swelling test and acrosome integrity with the fluorescein isothiocyanate-Pisum sativum agglutinin test. The semen was subsequently divided into four aliquots and diluted with Tris-egg yolk-glycerol (TEG), Tris-egg yolk without glycerol (TE), LDL with glycerol (LDL 1 ) and LDL without glycerol (LDL 2 ), at 80310 6 and 240310 6 sperm per ml. This study showed that the LDL 1 and LDL 2 extenders were more effective at preserving spermatozoa motility, plasma membrane integrity and acrosome integrity than TEG and TE (P,0.05) during 8 days of incubation. After 3 days of incubation, a toxicity of glycerol was observed in TEG, whereas no significant difference was observed between LDL 1 and LDL 2 . We can therefore conclude that the LDL extender can be used to refrigerate semen at 4 6C instead of TEG and TE at 80310 6 and 240310 6 sperm per ml for elite bulls. This finding can be used to define a policy for the storage of high-quality bull semen.

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“…The higher volume of seminal plasma would seem to be beneficial for cryopreservation, 11 but not for refrigeration at 4 uC for more than 3…”

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Effect of low-density lipoproteins, spermatozoa concentration and glycerol on functional and motility parameters of bull spermatozoa during storage at 4 °C

Vera-Munoz¹,

Amirat-Briand²,

Bencharif³

et al. 2010

Asian J Androl

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“…In most studies, substitution of egg yolk with 8% LDL improved the qualitative parameters of frozen-thawed spermatozoa immediately after thawing in comparison to extenders with whole egg yolk (Moussa et al 2002;Hu et al 2010Hu et al , 2011Stadnik et al 2015) and soy bean lecithin-based doi: 10.17221/27/2016-CJAS extenders as well (Amirat et al 2005;Vera-Munoz et al 2009). …”

Section: Discussionmentioning

confidence: 99%

“…Different cryoprotective properties of LDL, that positively and directly affected spermatozoa structure (Bergeron et al 2004) and/or extracellular conditions, were found (Hu et al 2011). The positive effects of egg yolk substitution by LDL in semen extenders on qualitative parameters of spermatozoa have been repeatedly demonstrated (Moussa et al 2002;Vera-Munoz et al 2009;Amirat-Briand et al 2010;Hu et al 2010Hu et al , 2011. The question then arises on whether the cryoprotective properties of soybean lecithinbased extenders could be improved by LDL addition.…”

Section: Introductionmentioning

confidence: 99%

Effect of low-density lipoprotein addition to soybean lecithin-based extenders on bull spermatozoa following freezing-thawing - preliminary results

Šimoník¹,

Rajmon²,

Stádník³

et al. 2016

Czech J. Anim. Sci.

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Soybean lecithin-based extenders represent an alternative to extenders containing egg yolk, but there has been discussion about their cryoprotective efficacy. Low-density lipoprotein (LDL) was proved as a cryoprotective agent, which can replace egg yolk in extenders. The aim of this study was to investigate the effects of LDL addition to the soybean extenders on their cryoprotective properties. The effect of the LDL of our production was verified using commercial egg yolk extender BULLXcell ® , 6%, 8%, and 10% LDL (v/v) as an egg yolk replacement. The effects of LDL addition to the soybean lecithin-based extenders in concentrations of 4%, 6%, and 8% (v/v) were tested using extenders AndroMed ® and Bioxcell ® . In total, 64 samples from eight bulls were evaluated. Kinematic parameters of spermatozoa, resulting from Computer Assisted Sperm Analysis, and their viability, evaluated by fluorescent technique, were assessed immediately after thawing and after 2 hours. The quality of LDL compared to other studies was confirmed, and the beneficial effects of egg yolk replacement by LDL were proved in extender BULLXcell ® . 8% LDL provided the best values for the majority of kinematic parameters (P < 0.05), without effect on total motility (P > 0.05). Furthermore, addition of 4%, 6%, and 8% LDL to the soybean lecithin-based extender Bioxcell ® showed a positive effect on the majority of kinematic parameters of spermatozoa (P < 0.05) at both times of incubation. However there was no significant influence on total motility (P > 0.05). Viability was higher after thawing in the case of 8% LDL (P < 0.05). However, there was no consistent effect of LDL addition to the AndroMed ® extender. In conclusion, cryoprotective properties of the semen extenders based on a soybean lecithin can be improved by the addition of LDL.

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“…Isto se torna mais problemá-tico quando se trata de amostras congeladas/descongeladas. Siqueira et al (2007), Borges et al (2011) e Oliveira et al (2012 utilizaram 20µL de sêmen congelado/descongelado em 1mL de solução hiposmótica, enquanto Vera-Munoz et al (2009) utilizaram 100µL de sêmen congelado/descongelado em 1mL de solução hiposmótica. Outros autores utilizaram diferentes proporções de sêmen congelado/descongelado e solução hiposmótica, Tuncer et al (2011) utilizaram 250µL de sêmen em 300 µL de solução hiposmótica e Nava--Trujillo et al (2011) 30µL de sêmen em 300µL de solução hiposmótica.…”

Section: Introductionunclassified

Utilização De Diferentes Volumes De Sêmen Na Realização Do Teste Hiposmótico Em Touros Adultos Da Raça Nelore

Martins

Pinho

Guimarães

et al. 2015

ArqVet

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MARTINS, L. F.; PINHO, R. O.; GUIMARÃES, J. D.; MACEDO, G. G.; BOSCARATO, A. G.; RODRIGUES, D. P.; DIAS, S. H. Utilização de diferentes volumes de sêmen na realização do teste hiposmótico em touros adultos da raça Nelore. Arq. Ciênc. Vet. Zool. UNIPAR, Umuarama, v.18, n.1, p. 5-10, jan./mar. 2015. RESUMO:O objetivo deste estudo foi avaliar diferentes proporções de sêmen: solução hiposmótica na realização do teste hiposmótico e suas relações com a congelabilidade do sêmen de touros zebuínos. Utilizaram-se 15 ejaculados de três touros adultos da raça Nelore. No sêmen in natura realizou-se a avaliação física e morfológica, a coloração supravital e o teste hiposmótico. No teste hiposmótico foi utilizada uma solução com osmolaridade de 100 mOsm/Kg com 15 minutos de perí-odo de incubação a 37 ºC, tanto no sêmen in natura quanto no congelado/descongelado. Foram utilizados quatro volumes de sêmen em 1mL de solução hiposmótica: 10, 20, 50 e 100 µL. As amostras criopreservadas foram descongeladas e foram realizados os testes hiposmótico, coloração supravital, teste de termo-resistência lento e a coloração fluorescente. Os valores médios e desvios padrão do percentual de espermatozoides reativos ao teste hiposmótico em sêmen in natura e congelado/ descongelado foram 69,3 ± 11,8 e 20,5 ± 6,8; respectivamente. Não houve correlação do teste hiposmótico com os aspectos físicos e morfológicos e os testes complementares realizados em sêmen in natura e congelado/descongelado. Nenhum teste de integridade de membrana plasmática dos espermatozoides foi capaz de classificar os touros quanto a sua congelabilidade do sêmen. Conclui-se que o teste hiposmótico pode ser realizado com 20 a 100 µL de sêmen in natura, e 10 a 100 µL de sê-men congelado/descongelado em 1 mL de solução hiposmótica, sem interferir em seus resultados, mas deve-se optar por 100 µL tanto para sêmen in natura e congelado/descongelado, porque melhora consideravelmente a leitura das lâminas. PALAVRAS-CHAVE: Bos indicus. Criopreservação. Macho. Testes Complementares. USE OF DIFFERENT SEMEN VOLUMES IN HYPOOSMOTIC TEST OF ADULT NELORE BULLSABSTRACT: The objective of this study was to evaluate different proportions of semen: hypoosmotic solution in the hypoosmotic swelling test and their relationship with semen freezability in Zebu bulls. A total of 15 ejaculates from three adult Nelore bulls were used. Physical and morphological features were analyzed in fresh semen, as well as supravital staining and hypoosmotic swelling test. In the hypoosmotic test, a hypoosmotic solution with 100 mOsm/kg osmolality using 15 minutes incubation at 37 °C was used both in fresh and frozen/thawed semen. Four semen volumes in 1-ml hyposmotic solution were used: 10, 20, 50 and 100 µL. Cryopreserved samples were thawed and submitted to hypoosmotic tests, supravital staining, slow thermo-resistance test and fluorescent staining. Mean values and standard deviations of the percentage of reactive sperm cells in the hypoosmotic test in fresh and frozen/thawed semen were 69.3 ± 11.8 and 20.5 ± 6.8...

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Effect of semen dilution to low-sperm number per dose on motility and functionality of cryopreserved bovine spermatozoa using low-density lipoproteins (LDL) extender: Comparison to Triladyl® and Bioxcell®

Cited by 42 publications

References 22 publications

Effect of low-density lipoproteins, spermatozoa concentration and glycerol on functional and motility parameters of bull spermatozoa during storage at 4 °C

Effect of low-density lipoproteins, spermatozoa concentration and glycerol on functional and motility parameters of bull spermatozoa during storage at 4 °C

Effect of low-density lipoprotein addition to soybean lecithin-based extenders on bull spermatozoa following freezing-thawing - preliminary results

Utilização De Diferentes Volumes De Sêmen Na Realização Do Teste Hiposmótico Em Touros Adultos Da Raça Nelore

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