Amides as cryoprotectants for freezing stallion semen: A review

Alvarenga, Marco Antônio; Papa, Frederico Ozanam; Landim-Alvarenga, F. C.; Medeiros, A.R.C

doi:10.1016/j.anireprosci.2005.07.001

Cited by 169 publications

(156 citation statements)

References 13 publications

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“…Medeiros et al (2002) comparando crioprotetores com diferentes concentrações de amida (3 a 5%) e a base de glicerol observaram motilidade total de 49 a 52% e motilidade progressiva de 10 a 15% para os a base de amida e de 27% e 8%, respectivamente, para os a base de glicerol. Segundo Alvarenga et al (2005) somente glicerol, provavelmente devido à melhor permeabilidade das aminas na célula espermática e, portanto, a menores injúrias espermáticas (Alvarenga & Papa 2011). A porcentagem média de anormalidades morfológicas maiores, menores e totais foram significativamente menores que os valores limites recomendados pelo CBRA (2013), indicando que o sêmen dos animais poderia ser comercializado.…”

Section: Discussionunclassified

“…Embora apresente inúmeras vantagens, a criopreservação apresenta limitações em várias raças equinas (Alvarenga et al 2004), que geram um entrave à sua disseminação (Brinsko et al 2000, Batellier et al 2001, Carmo & Almeida 2006. Isto porque o congelamento/descongelamento do sêmen ocasiona prejuízos, muitas vezes irreversíveis, aos espermatozoides como perda de motilidade, alterações estruturais e funcionais da membrana espermática e, consequentemente, diminuição do poder fecundante.…”

Section: Introductionunclassified

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Características do sêmen a fresco e descongelado de garanhões da raça Nordestina

et al. 2015

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RESUMO.-Este estudo descreveu as características seminais, da membrana plasmática e do acrossoma de espermatozoide congelado/descongelado de 19 ejaculados de garanhões da raça Nordestina. Os aspectos analisados incluíram os parâmetros físicos do sêmen fresco; a motilidade e a longevidade do sêmen diluído e descongelado; a morfologia espermática, integridade funcional e estrutural da membrana plasmática do espermatozoide e a habilidade de ligação do espermatozoide à membrana perivitelina da gema do ovo de galinha do sêmen descongelado. This paper describes the seminal characteristics of the plasma membrane of frozenthawed sperm. Nineteen ejaculates of Nordestino horse breed. The aspects analyzed in the physical parameters of fresh semen were total and progressive motility and your longevity after dilution or thawed; sperm morphology, functional and structural integrity of the plasma membrane of the sperm and the sperm-binding ability to the perivitelline membrane of the yolk (MPV) after thawed. The variables were assessed by ANOVA with post hoc test of Student Newman-Keuls test (P<0.05). The total and progressive motility were higher in diluted semen than thawed (P<0.05). The average percentage of the major, minor and total defects was lower than the limit recommended by the CBRA. The percentage of reactive to hypo-osmotic swelling test was 14.21±1.12%, the intact membrane detected by supravitally test was 62.22±9.06% and the SYBR-14 was 81.47±26.9. The ability of sperm to bind to the MPV after thawing semen was 230.39±57.09. The total and progressive motility at time 0 min of termo resistance test was higher than to 150 minutes (P<0.05), and no difference was observed in the times 10 and 30 minutes. The results demonstrate that the use of additional laboratory tests help in the process of evaluation of samples, making possible to obtain more reliable and accurate information. Although cryopreservation has caused decrease in sperm motility and was used diluents with amides to diluted and cryopreservation protocol and this minimized the osmotic damage to sperm cells and maintained the morphological, functional and structural integrity of the plasma membrane of the sperm. These results are a reference for future studies since there are no comparative data on this breed.

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Section: Discussionunclassified

Section: Introductionunclassified

Características do sêmen a fresco e descongelado de garanhões da raça Nordestina

et al. 2015

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“…Sin embargo, se ha sugerido que la dosis tóxica de GLY en espermatozoides de asno parece ser casi la mitad que dicha dosis en espermatozoides equinos, además de comprometer la fertilidad del semen congelado (Vidament et al, 2009). Entre las principales alternativas, la dimetilformamida (DMF) es conocida por su efecto crioprotector en semen equino, que se atribuye a su menor toxicidad y su mayor capacidad de difusión a través de la membrana de los espermatozoides (Alvarenga et al, 2005), debido a su menor peso molecular. En un estudio reciente, se describió un efecto positivo de la DMF sobre la movilidad y la integridad de membrana de espermatozoides criopreservados de la raza asnal andaluza (Acha et al, 2015).…”

Section: Introductionunclassified

Efecto de la suplementación del diluyente sobre la calidad del semen de asno a la descongelación

2017

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Palabras clave adicionalesCalidad seminal. Crioprotectores. Plasma seminal. Yema de huevo. Criopreservación. resUMenLa congelación de semen de asno permite almacenar material genético por un periodo ilimitado de tiempo, por lo que puede ser empleado para generar bancos de germoplasma o para la cría de mulares y asnos. Esta investigación tuvo como objetivo evaluar alternativas de suplementación de un diluyente para la congelación de semen de asno. Se colectó el semen de cinco asnos criollos colombianos, dos eyaculados por animal (n= 10) y luego se congeló en diluyente Equiplus® suplementado con 3% de glicerol (GLY) ó 5% de dimetilformamida (DMF), 2% ó 5% de yema de huevo centrifugada (YHC) y 10% ó 20% de plasma seminal (PS). Se realizó la evaluación post-descongelación de la calidad seminal y de la peroxidación lipídica (PL). Para el análisis estadístico se ajustaron modelos mixtos, se realizó un análisis de regresión y comparaciones de medias por la prueba de Tukey. El semen congelado con 5% de DMF, 5% de YHC y 10% de PS presentó valores superiores de movilidad, vitalidad e integridad de membrana (p<0,05). La PL fue menor en diluyentes con 5% de YHC (p<0,05). La suplementación del diluyente de congelación con una combinación de 5% de DMF, 5% de YHC y 10% de PS permite mejorar la calidad post-descongelación del semen de asno. Effect of extender supplementation on the post-thaw semen quality of donkey sUMMarY Freezing of donkey semen allows storing genetic material for an unlimited period of time, which can be used to generate germoplasm banks or for breeding of mules or asses. This research aimed to evaluate supplementation alternatives for freezing donkey semen. Semen from five Colombian creole donkeys (n= 10) was collected and then frozen in Equiplus® extender supplemented with 3% of glycerol (GLY) or 5% of dimethylformamide (DMF), 2% or 5% of centrifuged egg yolk (CEY) and 10% or 20% of seminal plasma (SP). Post-thaw evaluation of semen quality and lipid peroxidation (LP) was conducted. For statistical analysis mixed models were adjusted, regression analysis and comparison of means by the Tukey test were performed. Frozen semen with DMF, 5% CEY and 10% SP presented superior results of motility and membrane integrity (p<0.05). The LP was lower in extenders with 5% CEY (p<0.05). Supplementation of freezing extender with the combination of 5% DMF, 5% CEY and 10% SP allows to improve the post-thaw quality of asinine semen.additional keYwords

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“…Glycerol was used successfully as the first cryoprotectant for spermatozoa (Polge et al 1949) and the first pregnancy using frozen semen was reported by Baker and Gandier (1957). The effect of glycerol is not only protective but also toxic (Marcias Garcia et al 2012) and this could be a cause for the variation of stallion sperm motility after thawing (Alvarenga et al 2005). Contraceptive effects of glycerol in mares has also been published (Demick et al 1976).…”

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Effects of glycerol concentration on the motility of equine spermatozoa after thawing

et al. 2017

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The aim of this study was to evaluate the effect of different glycerol concentrations on stallion sperm motility after thawing. For statistical analysis 228 ejaculates were used. The semen was filtrated to remove gel fraction; macroscopic and microscopic evaluation was done. After evaluation the ejaculates were centrifuged, the supernatant was removed and the spermatozoa were re-suspended in French diluent with different concentrations of glycerol (2.0; 2.5; 4.0 and 6.0%). The choice of concentration of glycerol for a particular ejaculate was completely random. The spermatozoa were packed into 0.5 ml straws and placed for 2 h in a fridge (4 °C). Then the straws were placed in liquid nitrogen vapor (-80 to -100 °C) and after 10 min plunged into liquid nitrogen and stored at -196 °C for at least 48 h. The selected straws were individually thawed in a 38 °C water bath for 30 s prior to post-freezing analysis. Two progressive motilities using phase contrast microscopy (magnification × 400) were recorded: motility II immediately after thawing and motility III after 2 h incubation in a 38 °C water bath. The Spearmen/Kendall rang correlation test was selected to prove whether there is a correlation between the selected indices (glycerol concentration and motility II and motility III). Nonparametric multiple group analysis (Steel-Dwass test) was applied for finding the differences between groups. The Spearman/Kendall rang correlation proved a relationship between motility II and glycerol concentration. It can be stated that in this study the best glycerol concentration for freezing equine spermatozoa is with a concentration of 4.0% glycerol.

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Amides as cryoprotectants for freezing stallion semen: A review

Cited by 169 publications

References 13 publications

Características do sêmen a fresco e descongelado de garanhões da raça Nordestina

Características do sêmen a fresco e descongelado de garanhões da raça Nordestina

Efecto de la suplementación del diluyente sobre la calidad del semen de asno a la descongelación

Effects of glycerol concentration on the motility of equine spermatozoa after thawing

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