Ameliorative Effect of Ascorbic Acid and Glutathione in Combating the Cryoinjuries During Cryopreservation of Exotic Jack Semen

Kumar, Prashant; Kumar, Rabindra; Mehta, Jitendra; Chaudhary, Ashok; Ravi, S. K.; Mehta, S. L.; Ansari, Md. Meraj; Legha, Ram Avtar; Tripathi, B. N.

doi:10.1016/j.jevs.2019.102796

Cited by 12 publications

(13 citation statements)

References 62 publications

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“…While sperm viability and integrities of plasma and acrosome membranes were not affected by the presence of high GSH concentrations, no treatment, even the ones containing low concentrations of this antioxidant, improved these sperm parameters post-thaw. This is contrary to what was found by Kumar et al [16] in frozen-thawed sperm from exotic donkeys (Poitou breed, Martina Franca), where there was a significant increase in motility, viability and plasma membrane integrity when the freezing medium was supplemented with 2.5 mM. Our results, however, agree with the aforementioned study in the percentage of sperm with an intact acrosome membrane, since no differences in this parameter were observed in our study or in that of Kumar et al [16], even when using two different techniques (flow cytometry vs. Giemsa stain + microscopy).…”

Section: Discussioncontrasting

confidence: 94%

“…This is contrary to what was found by Kumar et al [16] in frozen-thawed sperm from exotic donkeys (Poitou breed, Martina Franca), where there was a significant increase in motility, viability and plasma membrane integrity when the freezing medium was supplemented with 2.5 mM. Our results, however, agree with the aforementioned study in the percentage of sperm with an intact acrosome membrane, since no differences in this parameter were observed in our study or in that of Kumar et al [16], even when using two different techniques (flow cytometry vs. Giemsa stain + microscopy). Moreover, comparing our results with those found in other species may provide some clues, despite the fact that inconsistent data are seen in the literature.…”

Section: Discussioncontrasting

confidence: 94%

“…The same effect has been observed in pigs, as the addition of 1 mM GSH to the freezing medium results in greater protection of sperm function than higher concentrations [41]. In donkeys, there is only one study that investigated the effects of supplementing the freezing medium with GSH; however, only one concentration was tested [16]. Testing the resilience of donkey sperm to oxidative stress using different GSH concentrations could shed some light into the reproductive strategy of this species [50][51][52].…”

Section: Introductionmentioning

confidence: 67%

“…This last aspect is very relevant in the donkey because the size of the population has suffered a significant decrease in recent years [4], which directly affects the genetic variability of the existing breeds [5]. Research in this species has focused its efforts on improving sperm cryopreservation protocols, precisely to try to safeguard endangered donkey breeds [6][7][8][9][10][11][12][13][14][15][16][17][18]. However, the use of cryopreserved donkey sperm has been limited by the low conception rates obtained when jennies are inseminated [7,8,19].…”

Section: Introductionmentioning

confidence: 99%

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Addition of Reduced Glutathione (GSH) to Freezing Medium Reduces Intracellular ROS Levels in Donkey Sperm

Yánez-Ortiz

Catalán

Delgado-Bermúdez

et al. 2021

Veterinary Sciences

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In donkeys, the use of frozen-thawed sperm for artificial insemination (AI) leads to low fertility rates. Furthermore, donkey sperm produce a large amount of reactive oxygen species (ROS), and post-AI inflammation induces the formation of neutrophil extracellular traps (NETosis), which further generates many more ROS. These high ROS levels may induce lipid peroxidation in the sperm plasma membrane, thus affecting its integrity. Enzymatic and non-enzymatic antioxidants, mainly found in the seminal plasma (SP), are responsible for maintaining the redox balance. However, this fluid is removed prior to cryopreservation, thereby exposing sperm cells to further oxidative stress. The exogenous addition of antioxidants to the freezing medium can reduce the detrimental effects caused by ROS generation. Therefore, the aim of this study was to evaluate how the addition of different reduced glutathione (GSH) concentrations (control, 2 mM, 4 mM, 6 mM, 8 mM, and 10 mM) to fresh sperm affect their cryotolerance. Total and progressive motility, kinematic parameters and motile sperm subpopulations were significantly (p < 0.05) different from the control in treatments containing 8 mM and 10 mM GSH, but not at lower concentrations. Plasma and acrosome membrane integrity, mitochondrial membrane potential (MMP) and intracellular superoxide levels (O2−) were not affected (p > 0.05) by any GSH concentration. Interestingly, however, the addition of 8 mM or 10 mM GSH reduced (p < 0.05) the percentages of viable sperm with high overall ROS levels compared to the control. In conclusion, frozen-thawed donkey sperm are able to tolerate high GSH concentrations, which differs from what has been observed in other species. This antioxidant capacity suggests that ROS could be important during post-AI and that the impact of using exogenous antioxidants like GSH to improve the sperm resilience to freeze-thawing is limited in this species.

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Section: Discussioncontrasting

confidence: 94%

Section: Discussioncontrasting

confidence: 94%

Section: Introductionmentioning

confidence: 67%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Addition of Reduced Glutathione (GSH) to Freezing Medium Reduces Intracellular ROS Levels in Donkey Sperm

Yánez-Ortiz

Catalán

Delgado-Bermúdez

et al. 2021

Veterinary Sciences

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“…Cryopreservation of semen is an important biotechnological tool, which is useful for the conservation of biodiversity of donkey species (Kumar et al 2018). However, the results obtained in donkey semen cryopreservation were found highly variable (Legha et al 2013, Prashant Kumar et al 2019. Most of the procedures used for cryopreservation of donkey semen have been adopted from those reported in horse stallions.…”

Section: Resultsmentioning

confidence: 99%

Supplementation of caffeine to extender improves post-thaw seminal attributes of Poitou donkey semen

Kant

Pal

Legha

et al. 2021

Indian J of Anim Sci

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The current study was conducted with an objective of elucidating the effect of addition of various levels of caffeine to jack semen on seminal attributes at pre- and post-thaw stage. Six ejaculates from each three adult jack stallions were collected via artificial vagina using estrus jenny as dummy. Caffeine was added to semen extender at 0.1, 0.5 and 2 mM at pre-freeze stage and the semen was cryopreserved according to standard protocol. Sperm motility, viability and other functional attributes were assessed at pre-freeze and post-thaw stages. The present study revealed that, adding caffeine to jack semen extender at 0.1 mM, 0.5 mM or 2 mM levels had positive impact on seminal parameters compared to the control semen. Sperm kinetics (VAP, VSL and STR) and motility increased in the caffeine treated groups in comparison to the control group. The caffeine at 2.0 mM concentration resulted in better post-thaw sperm motility, viability, and acrosome integrity as compared to 1 or 0.5 mM of caffeine and control. Caffeine supplementation also enhanced post-thaw sperm kinematic parameters, which implies its potential as an alternative antioxidant supplement. It was concluded that, 2 mM caffeine supplementation to donkey semen significantly improved post-thaw sperm motility and other functional attributes as compared to the control. However, the potential benefits of caffeine on the fertility rates needs to be investigated.

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Cholesterol Loaded Cyclodextrin Supplementation Enhances the Cholesterol-to-Phospholipid Ratio and Diminishes Oxidative Stress in Jack Spermatozoa During Cryopreservation

Kumar

Mehta

Ravi

et al. 2020

Journal of Equine Veterinary Science

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Ameliorative Effect of Ascorbic Acid and Glutathione in Combating the Cryoinjuries During Cryopreservation of Exotic Jack Semen

Cited by 12 publications

References 62 publications

Addition of Reduced Glutathione (GSH) to Freezing Medium Reduces Intracellular ROS Levels in Donkey Sperm

Addition of Reduced Glutathione (GSH) to Freezing Medium Reduces Intracellular ROS Levels in Donkey Sperm

Supplementation of caffeine to extender improves post-thaw seminal attributes of Poitou donkey semen

Cholesterol Loaded Cyclodextrin Supplementation Enhances the Cholesterol-to-Phospholipid Ratio and Diminishes Oxidative Stress in Jack Spermatozoa During Cryopreservation

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