Efecto de Tres Crioprotectores en la Criopreservación de Espermatozoides Epididimarios de Alpaca

Terreros, C Marino; Huanca, L Wilfredo; Arriaga, C Irma; Ampuero, B Antonio

doi:10.15381/rivep.v26i3.11182

Cited by 5 publications

(6 citation statements)

References 14 publications

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“…The height was graduated manually, remaining 10 min at 24 cm, 10 min at 12 cm, and 5 min at 3 cm above the liquid nitrogen level. Finally, the straws were plunged into the liquid nitrogen and stored in a nitrogen tank at −196°C [modified from ( 30 )].…”

Section: Methodsmentioning

confidence: 99%

Spermatozoa Obtained From Alpaca vas deferens. Effects of Seminal Plasma Added at Post-thawing

et al. 2021

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The viscous seminal plasma (SP) is currently a major impediment to the handling of ejaculate and the development of some biotechnologies in South American camelids. The vas deferens-collected spermatozoa of alpacas is a useful technique to avoid this problem. On the other hand, SP contains a large protein component that has been implicated in the function of spermatozoa within the female reproductive tract. In this sense, the low fertility achieved using transcervical insemination with frozen-thawed spermatozoa in alpacas could be improved by adding SP. This study aimed to evaluate the effect of the whole SP on some in vitro parameters of alpaca spermatozoa after the freezing-thawing-process and the fertility after artificial insemination. It would contribute to a better understanding of the interaction between thawed sperm cells and SP. Spermatozoa were obtained by surgically diverted vas deferens. The samples were diluted with a Tris-based extender, packaged in straws, and frozen. At thawing, each straw was divided into two post-thawing conditions: with the addition of 10% of PBS (control) or with 10% SP (treatment). The sperm cells were evaluated using dynamic parameters, sperm cell morphology, and morphometry. Fertility was assessed by an artificial insemination trial. All in vitro parameters were analyzed by ANOVA. A heterogeneity test was scheduled for the fertility trial. After the freezing-thawing process, motility and plasma membrane functionality was improved when SP was added. No differences were found for post-thaw viability between the control and treatment samples. The percentage of normal cells was higher with SP at post-thawing, and a decrease of the presence of bent tailed spermatozoa with a droplet in the SP group was observed. The length of the head spermatozoa was 3.4% higher in the samples with PBS compared to those in which SP was added. Females pregnant at day 25 post-insemination were 0/12 (with SP inside the straw) and 1/10 (without SP inside the straw). In conclusion, the presence of 10% SP at post-thawing improves sperm cells' motility, functionality, and morphology, indicating that it would be beneficial to improve the frozen-thawed alpaca's physiology spermatozoa. More fertility trials must be developed to increase this knowledge.

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Section: Methodsmentioning

confidence: 99%

Spermatozoa Obtained From Alpaca vas deferens. Effects of Seminal Plasma Added at Post-thawing

et al. 2021

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“…Esto podría deberse al dilutor utilizado en ambos estudios (Tris) el cual no contenía proteínas de la leche y lipoproteínas de baja densidad (LDL), que se han demostrado protegen a los espermatozoides del choque térmico y mejoran la motilidad espermática entre otros parámetros (19). Por otro lado, aunque otros autores utilizaron el mismo dilutor (a base de leche descremada, yema de huevo y fructuosa) también reportaron motilidades menores (46.1±7.7%; 46.1±7.7% y 62.5±19.4%) (10,11,22), lo cual podría deberse a que existen diferencias en los parámetros espermáticos entre individuos y dentro de ellos. (23).…”

Section: Discusionunclassified

“…En alpacas, durante la criopreservación de espermatozoides, se observa un aumento de las ROS (8), lo que induce una disminución de la motilidad, de la integridad de la membrana plasmática y acrosomal, de la actividad mitocondrial, de la viabilidad y un aumento en la fragmentación de ADN (9,10,11,12). Se ha reportado que concentraciones de PS al 10% en esperamtozoides epididimarios de alpaca, logran mantener la motilidad e integridad acrosomal y reducen el porcentaje de espermatozoides muertos (12).…”

Section: Introductionunclassified

Oxidative stress on alpaca epididimal sperm frozen with different concentrations of seminal plasma

Jimenez-Carpio,

Ugarelli-Galarza,

Evangelista-Vargas

2024

Rev MVZ Córdoba

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Objective. To assess the oxidative stress on alpaca epididimal sperm frozen with different concentration of seminal plasma. Materials and methods. 29 post mortem alpaca epididymis were used. Epididymal sperm were obteined thought cuts and suspended in a diluent based on skim milk, egg yolk and fructose and dimethylacetamine. Samples were separated into four aliquots which were supplied with seminal plasma (v/v) in concentrations of 0, 10, 25 and 50% respectively; then, sperm motility was assessed before freezing; then, motility was assessed before freezing with an automatic freezer machine. After thawing, mitochondrial membrane activity, sperm viability, lipid peroxidation and sperm apoptosis were assessed using flow cytometry with images. Results. Sperm exposed to 50% seminal plasma showed the lowest post-thaw motility (3.5±3.7%) and the highest percentages of apoptotic cells (71.43±20.6%). No significant differences were found in the rest of parameters. Conclusions. This study constitutes non-conclusive evidence that presence of seminal plasma does not affect viability, mitochondrial activity, or lipid peroxidation of alpaca epididymal sperm during cryopreservation processes; however, when it is absent or added up to 25% in the cryopreservation process, it may be possible to obtain better results in terms of sperm motility and sperm apoptosis.

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“…1,2 Later work testing alternative cryprotectants demonstrated that post-thaw motility was improved when dimethylsulfoxide (DMSO) or diacetylamide (DMA) were used instead of ethylene glycol or glycerol. 3,4 With addition of these cryoprotectants, mean post-thaw motility ranged from 31 to 34%. 3,4 Amides, like DMA and dimethylformamide (DMFA) are small hydrophilic molecules with better cell penetration and less toxicity than glycerol.…”

Section: Introductionmentioning

confidence: 99%

Semen extender type and cold storage of tissue affect cryosurvival of alpaca epididymal sperm

Ferrer,

Williamson,

Palomares

et al. 2018

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The objectives of this study were to determine the ability of liposome- and DMFA-containing semen extenders to support cryosurvival of alpaca epididymal sperm, and to evaluate the effect of epididymal cold storage on post-thaw viability. It was hypothesized that both semen extenders provide appropriate cryoprotection, and post-thaw sperm viability does not differ when alpaca epididymal sperm are cryopreserved immediately after castration or after 24 h of cold storage. Sperm were recovered from one epididymis from each male (n = 10) on the day of castration, and from the other one after 24 h of tissue storage at 4C. On each day, each sample was divided into two aliquots, which were cryopreserved in semen extender OptiXcell or BotuCRIO. Pre-freezing sperm parameters did not differ between samples obtained on the day of castration or 24 h later. Motility of sperm cryopreserved in BotuCRIO on the day of castration did not differ from pre-freezing values. Use of OptiXcell or delaying sperm cryopreservation decreased total (p = 0.002) and progressive sperm motility (p = 0.041) compared with pre-freezing values seen in sperm recovered on the day of castration. Membrane integrity was lowest when sperm were cryopreserved in OptiXcell after cold storage. Treatment had no effect on DNA integrity. It was concluded that the DMFA-containing semen extender BotuCRIO was superior to the liposome-containing semen extender OptiXcell at preserving post-thaw viability of alpaca epididymal sperm. Cold storage of epididymides for 24 h negatively affected post-thaw motility and membrane integrity of sperm. Therefore, cryopreservation with BotuCRIO immediately after castration is recommended for best preservation of post-thaw viability of alpaca epididymal sperm.

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Efecto de Tres Crioprotectores en la Criopreservación de Espermatozoides Epididimarios de Alpaca

Cited by 5 publications

References 14 publications

Spermatozoa Obtained From Alpaca vas deferens. Effects of Seminal Plasma Added at Post-thawing

Spermatozoa Obtained From Alpaca vas deferens. Effects of Seminal Plasma Added at Post-thawing

Oxidative stress on alpaca epididimal sperm frozen with different concentrations of seminal plasma

Semen extender type and cold storage of tissue affect cryosurvival of alpaca epididymal sperm

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