Use of Equine Sperm Cryopreservation Techniques as a Conservation Method of Donkey Germplasm

Monteiro, Rodrigo Alves; Cunha, Rodolfo Marinho; Guerra, Maria Madalena Pessoa; Almeida, Valdir M.; Peña-Alfaro, Carlos Enrique; Silva, Sildivane Valcácia

doi:10.1016/j.jevs.2022.103888

Cited by 4 publications

(3 citation statements)

References 43 publications

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“…Seminal plasma removal is another divergent issue about successful sperm cryopreservation. One since of the main negative impacts on SP is the formation of reactive oxygen species (ROS; Monteiro et al, 2022). Quantities bigger than 10% SP in semen samples, in horses, cause negative effects on spermatozoa submitted cryopreservation and ROS participation is attributed in these cases (Len et al, 2020).…”

Section: Discussionmentioning

confidence: 99%

“…SP is a mixture of fluid secreted from the testes, epididymis and accessory sex glands, and it is involved in a multitude of sperm functions and events preceding fertilization (Kareskoski & Katila, 2008), such as activation of motility, antimicrobial action, neutralization of metabolites and a mediating effect of sperm capacitation and postcoital uterine inflammatory response (Rodríguez‐Martínez et al, 2011). Nevertheless, SP in Equidae must be removed for preservation processes of sperm in artificial conditions such as cryopreservation (Miró et al, 2013; Monteiro et al, 2022; Vilés et al, 2013), since has been associated with a deleterious effect on sperm capacitation and fertilization capacity (Morrell et al, 2014). Yet, some studies have reported promising results regarding the improvement of post‐thaw sperm quality when it is supplemented with small amounts of SP, which has been related to the contribution of SP to the antioxidant capacity of the cryopreserved semen (Usuga et al, 2020).…”

Section: Introductionmentioning

confidence: 99%

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Donkey semen cryopreservation: Alternatives with permeable, non‐permeable cryoprotectants and seminal plasma

Páez

Suarez

Betancur

2023

Reprod Domestic Animals

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Cryopreservation of semen is an important technique to preserve genetic material. Yet, pregnancy rates in jennies after artificial insemination with frozen–thawed donkey semen are poor. This condition has been attributed to the impact of permeable cryoprotectants, that could cause high post‐breeding endometritis. Removal of seminal plasma (SP) prior to semen freezing process is another contributing factor. SP is involved in a multitude of sperm functions and events preceding fertilization and has a mediating effect of sperm capacitation and postcoital uterine inflammatory response. The aim of this study was to evaluate different alternatives in donkey semen cryopreservation with permeable, non‐permeable cryoprotectants, BSA and SP. Thirty ejaculates from 10 donkeys were cryopreserved with different combinations of dimethylformamide (DMF, 5%), sucrose (SUC, 200 mM) and homologous SP (10%): DMF (T1), DMF/SP (T2), SUC/BSA (T3), SUC/BSA/SP (T4), DMF/SUC/BSA (T5), DMF/SUC/BSA/SP (T6), DMF/BSA (T7) and DMF/BSA/SP (T8). After thawing, sperm motility and kinetics were assessed by computerized semen analysis. Sperm vitality (SV) was evaluated by fluorescence microscopy, functional membrane integrity (FMI) by the HOST test, abnormal morphology by eosin‐nigrosin staining and sperm membrane stability by flow cytometry. For statistical analysis, sperm quality indexes (SQi) were obtained, general linear models were carried out and mean comparisons were made by the Tukey test. T1, T2, T5, T6, and T7 had higher and equivalent results for motility, most kinetic parameters and function membrane integrity. Cryopreservation of donkey semen without permeable cryoprotectant (T3 and T4) showed a reduction in motility, kinetics, SV, FMI and SQi. T5 showed a reduction in progressive motility, sperm velocities, IMF and SQi compared to other DMF treatments. T6 and T8 achieved higher SQi values compared to T1, but they were not different compared to T2 and T7. T1 had a smaller sperm population with low‐M540 compared to T3. It is concluded that the use of permeable cryoprotectant is essential to achieve higher post‐thaw quality of donkey semen. In addition, the combined use of BSA, SUC and/or PS may provide additional sperm protection compared to the individual use of DMF.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Donkey semen cryopreservation: Alternatives with permeable, non‐permeable cryoprotectants and seminal plasma

Páez

Suarez

Betancur

2023

Reprod Domestic Animals

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“…Cryopreservation was found to be successful in many mammalian species, whereas inefficient in porcine (Silva et al, 2015). This is due to the fact that boar sperm plasma membrane is different from that of the other animals; it consists of high poly-unsaturated fatty acid content and low concentrations of cholesterol (Yeste et al, 2017;Monteiro et al, 2022). As a result, cryopreservation process in boars may lead to changes of proteins and carbohydrate composition, which may disrupt membrane structure and reduce sperm viability (Silva et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

Sperm cryopreservation in Windsnyer boars; principles, technique, and updated outcomes

Thema,

Mphaphathi,

Ledwaba

et al. 2023

Anim. Reprod.

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The domestic pig breeds are in danger of extinction whereas the erosion of their gene pool is a serious concern because they significantly contribute to the rich biodiversity. Overall aim of this study was to determine the protocol for preserving the semen of the Windsnyer boars for conservation. A total of 18 ejaculates (6 replications/boar) were collected from three Windsnyer boars of proven fertility with the use of hand-gloved approach method, twice per week. Boars semen were pooled and extended with Beltsville Thawing Solution [(BTS) IMV Technologies, France], held at 18°C for 3 hours and centrifuged. The sperm pellet was re-suspended with Fraction A (20% egg yolk + BTS) and cooled at 5°C for 1 hour. Following cooling, semen was divided and diluted into different cryoprotectants (ethylene glycol, glycerol, propanediol, ethylene glycol + glycerol + propanediol) at equal contribution to make the total concentrations [4, 8, 12 and 16% and the 0% (control; without cryoprotectant)] and loaded into 0.25 mL straws. Two cryopreservation methods (liquid nitrogen vapour and controlled rated) were used to cryopreserve the semen straws. Semen straws were thawed at different temperatures (5, 18, 37 and 40°C) and evaluated for sperm motility, viability, and morphology traits. Post-thawed sperm total motility (36.0±5.3) and live normal sperm (49.5±8.3) percentages were recorded to be higher in the treatment supplemented with 16% glycerol (P<0.05). The highest sperm total motility percentage was recorded at 40°C (26.8±3.2) thawing temperature for liquid nitrogen vapour treatment (P<0.05). In conclusion, 16% glycerol was found to be the suitable cryoprotectant concentration for semen cryopreserved with liquid nitrogen vapour method and thawed at 40°C.

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ProAKAP4 as a motility long-lasting marker in Catalan donkey spermatozoa

Dordas-Perpinyà,

Sergeant,

Yánez-Ortiz

et al. 2024

Animal Reproduction Science

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Use of Equine Sperm Cryopreservation Techniques as a Conservation Method of Donkey Germplasm

Cited by 4 publications

References 43 publications

Donkey semen cryopreservation: Alternatives with permeable, non‐permeable cryoprotectants and seminal plasma

Donkey semen cryopreservation: Alternatives with permeable, non‐permeable cryoprotectants and seminal plasma

Sperm cryopreservation in Windsnyer boars; principles, technique, and updated outcomes

ProAKAP4 as a motility long-lasting marker in Catalan donkey spermatozoa

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