Evaluación De La Integridad De Adn Mediante Citometría De Flujo en Espermatozoides De Alpaca Criopreservados Con Análosos De Superóxido Dismutasa.

Santiani, Alexei; Evangelista, Virgilio; Cheuquemán, A Carolina; Baer, H Alejandra von; Risopatrón, G Jennie; Sánchez, G Raúl

doi:10.15381/rivep.v23i2.898

Cited by 5 publications

(4 citation statements)

References 4 publications

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“…One of the most important findings of this study is that no significant compromise on sperm chromatin condensation nor DNA fragmentation was detected with the tested freezing rate and the three final CPs concentrations compared to raw semen samples. These results are worth to mention since previous experiments detected a negative impact on DNA sperm quality of posthawed camelid sperm regardless of the CPs used (alpaca: Santiani et al, 2012;llama: Carretero, Neild, et al, 2015;Fumuso et al, 2019Fumuso et al, , 2020. observed a significant higher damage on frozenthawed llama sperm, with an increase of fragmented DNA when GL was used at 7%, and higher sperm chromatin decondensation with both GL and DMF (7%).…”

Section: (%) Lr (%) DI (%) Dr (%)supporting

confidence: 52%

“…Another difference is that in the present study, CPs were added progressively over a period of 15 min, exposing sperm to a gradual increase of CPs and avoiding sudden and possible toxic changes on the cells. As mention before, compromised DNA in frozen semen was also found in other camelids (alpaca: Santiani et al, 2012) but the authors successfully reduced this effect with the addition of tempol (analogue of superoxide dismutase) at 1 mM to the media during cryopreservation. This result is important to be considered as oxidative stress might also affect sperm characteristics such as sperm motility and DNA quality (Bansal & Bilaspuri, 2010;Fraczek & Kurpisz, 2005); thus, the addition of antioxidants could be a possible area of study to improve llama posthawed sperm parameters.…”

Section: (%) Lr (%) DI (%) Dr (%)mentioning

confidence: 90%

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Cryopreservation of llama semen using a combination of permeable cryoprotectants

Fumuso

Bertuzzi

González

et al. 2021

Reprod Domestic Animals

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Semen cryopreservation is not available for massive use in South American Camelids (SACs) due to the lack of an efficient protocol and the low pregnancy rates obtained with artificial insemination (AI). The use of a single cryoprotectant (CP) is commonly used in SACs frozen semen. The objective of the study was to evaluate the combined cryoprotective capacity of two permeable CPs at different stages of the cryopreservation protocol in llama semen. Sixteen ejaculates from 4 llama males were analysed, and sperm quality was assayed in raw semen, at 5°C, after equilibration of samples with the CPs and when samples were thawed. The following CPs and combination were used: 6% glycerol (GL), 6% dimethylformamide (DMF) and the combination of both CPs: 3% GL and 3% DMF. A Kruskal-Wallis test and an experimental factorial design, considering one factor with four levels (raw semen, 6% GL, 6% DMF and GL/DMF), were used. Total sperm motility and live sperm with intact acrosomes remained unchanged after equilibration of samples (p > .05). A significant decrease in the percentage of functional membrane, motile and live sperm with intact acrosomes was observed when samples were thawed (GL, DMF and GL/DMF). Nevertheless, the cryopreservation protocols used preserved sperm DNA quality; thus, sperm chromatin condensation and DNA fragmentation were unaffected (p > .05) when GL, DMF and GL/DMF were used. To conclude, no superiority was found between the use of a single or a combination of permeable cryoprotectants to freeze llama semen.

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Section: (%) Lr (%) DI (%) Dr (%)supporting

confidence: 52%

Section: (%) Lr (%) DI (%) Dr (%)mentioning

confidence: 90%

Cryopreservation of llama semen using a combination of permeable cryoprotectants

Fumuso

Bertuzzi

González

et al. 2021

Reprod Domestic Animals

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“…The evaluation was carried out using the commercial terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) kit (in situ cell death detection Kit, Fluorescein Roche ® Mannheim, BW, Germany). The sample's processing was performed as previously described with some modifications [38]. A sperm suspension of 2 × 10 6 spermatozoa /mL was fixed in 4% formaldehyde for 15 min at 4 • C. Subsequently, two washes were carried out with a phosphate saline solution (PBS).…”

Section: Dna Fragmentation Analysismentioning

confidence: 99%

Effect of Atomized Black Maca (Lepidium meyenii) Supplementation in the Cryopreservation of Alpaca (Vicugna pacos) Epididymal Spermatozoa

Sanchez

Quispe

Vargas

et al. 2023

Animals

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Artificial insemination is an important assisted reproductive technology that has been applied in several mammalian species. However, successful cryopreservation of semen of South American camelids has been limited, hindering the commercial application of artificial insemination in alpaca species. In this scenario, the addition of antioxidants to semen extenders provides a strategy to improve the freezability of mammalian sperm. Bioactive metabolites from natural extracts of black maca have shown valuable antioxidant properties. Thus, the objective of this study was to evaluate the effect of the addition of atomized black maca in the freezing medium of epididymal spermatozoa of alpacas. Fifteen pairs of epididymis were collected from a local slaughterhouse. Each sample was divided into six groups: (1) fresh, (2) yolk medium (YM), (3) 10 mg/mL maca, (4) 20 mg/mL maca, (5) 30 mg/mL maca, and (6) resveratrol (as an antioxidant control). Sperm cryopreservation was performed through the slow freezing method. Markers associated with functionality, such as motility, viability, and plasma membrane integrity, as well as markers associated with oxidative damage, such as DNA integrity, total ROS production, and mitochondrial function, were analyzed. The results show that the supplementation with black maca (20 mg/mL) improved the sperm motility, viability, plasma membrane integrity, and mitochondrial function evaluated according to an index of formazan deposits. Similarly, the ROS production decreased with maca at 20 mg/mL, although the DNA integrity did not show any differences among the groups. These results suggest that maca at 20 mg/mL has cytoprotective effects during freezing/thawing of epididymal sperm of alpaca species. Further research will be focused on assessing the effects of maca supplementation on semen extenders by using biomolecular markers (proAKAP4) associated with fertility.

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“…Entre otros, el antioxidante Tempol (4hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl), ha sido empleado para disminuir el efecto dañino del ROS, el cual previene la pérdida de motilidad y viabilidad espermática durante la refrigeración de los espermatozoides de ovino (Mara et al, 2005;Ruiz et al, 2007;Sandoval et al, 2007) (Santiani et al, 2005(Santiani et al, , 2012(Santiani et al, , 2013.…”

Section: Introductionunclassified

Adición del Antioxidante Tempol en la Criopreservación de Espermatozoides de Alpaca (Vicugna pacos) Colectados por Desviación del Conducto Deferente

Gómez-Quispe

Pérez

Machaca

2016

Rev. investig. vet. Perú

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El presente estudio tuvo como objetivo determinar el efecto de la adición del antioxidante Tempol en la motilidad individual y progresiva a la refrigeración y descongelación de espermatozoides de alpaca. Se desvió el conducto deferente de tres alpacas con adecuados parámetros seminales y los espermatozoides fueron colectados tres veces por semana y mezclados para anular el efecto individual. La muestra colectiva se dividió en cuatro alícuotas y fueron colocadas en refrigeración hasta alcanzar 5 ºC en 2 h. El antioxidante fue añadido en concentraciones de 0, 0.5, 1.0 y 1.5 mM cuando las alícuotas alcanzaron los 10 ºC. Las muestras fueron envasadas en pajillas de 0.25 ml, colocadas sobre vapores de nitrógeno líquido por 6 min y sumergidas. La motilidad individual de refrigeración fue 55.1, 63.7, 52.8 y 45.0 (p<0.05) y a la descongelación 35.7, 42.3, 29.0 y 18.1% (p<0.05) para 0, 0.5, 1.0 y 1.5 mM de Tempol, respectivamente. La motilidad progresiva a la refrigeración fue 36.0, 42.4, 28.4 y 21.7 (p<0.05) y a la descongelación 24.4, 27.3, 21.1 y 15.8% para 0, 0.5, 1.0 y 1.5 mM de Tempol, respectivamente (p<0.05). La adición de una baja concentración de antioxidante Tempol mejoró la motilidad individual y progresiva de la criopreservación de espermatozoides provenientes del conducto deferente de la alpaca.<br /><br />

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Evaluación De La Integridad De Adn Mediante Citometría De Flujo en Espermatozoides De Alpaca Criopreservados Con Análosos De Superóxido Dismutasa.

Cited by 5 publications

References 4 publications

Cryopreservation of llama semen using a combination of permeable cryoprotectants

Cryopreservation of llama semen using a combination of permeable cryoprotectants

Effect of Atomized Black Maca (Lepidium meyenii) Supplementation in the Cryopreservation of Alpaca (Vicugna pacos) Epididymal Spermatozoa

Adición del Antioxidante Tempol en la Criopreservación de Espermatozoides de Alpaca (Vicugna pacos) Colectados por Desviación del Conducto Deferente

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