Effect of the holding time at 15°C prior to cryopreservation, the thawing rate and the post-thaw incubation temperature on the boar sperm quality after cryopreservation

Abstract Holding at room temperature is the first step in most boar semen cryopreservation protocols. It is well accepted that a holding time (HT) of 24 h increases sperm cryotolerance. However, the effect of HT on ejaculates with different freezability is not entirely clear. The aim of this study was to understand how HT influences spermatic and seminal plasma metabolite profiles of boar ejaculates and how these possible changes affect freezability. Twenty-seven ejaculates were collected and extended to 1:1 (v: v) with BTS and split into two aliquots. The first aliquot was cryopreserved without holding time (0 h), and the second was held at 17°C for 24 h before cryopreservation. Spermatozoa and seminal plasma were collected by centrifugation at two times, before HT (0 h) and after HT (24 h), and subsequently frozen until metabolite extraction and UPLC–MS analysis. After thawing, the semen samples were evaluated for kinetics, membrane integrity, mitochondrial potential, membrane lipid peroxidation, and fluidity. The ejaculates were then allocated into two phenotypes (good ejaculate freezers [GEF] and poor ejaculate freezers [PEF]) based on the percent reduction in sperm quality (%RSQ) as determined by the difference in total motility and membrane integrity between raw and post-thaw samples cryopreserved after 24 h of HT. The metabolic profile of the seminal plasma did not seem to influence ejaculate freezability, but that of the spermatozoa were markedly different between GEF and PEF. We identified a number of metabolic markers in the sperm cells (including inosine, hypoxanthine, creatine, ADP, niacinamide, spermine, and 2-methylbutyrylcarnitine) that were directly related to the improvement of ejaculate freezability during HT; these were components of metabolic pathways associated with energy production. Furthermore, PEF showed an up-regulation in the arginine and proline as well as the glutathione metabolism pathways. These findings help to better understand the effect of holding time on boar sperm freezability and propose prospective metabolic markers that may predict freezability; this has implications in both basic and applied sciences.

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Effects of iodine methionine on boar sperm quality during liquid storage at 17°C

Reproduction in Domestic Animals ◽

10.1111/rda.13024 ◽

2017 ◽

Vol 52 (6) ◽

pp. 1061-1066 ◽

Cited By ~ 5

Author(s):

Q Fang ◽

J Wang ◽

YY Hao ◽

H Li ◽

JX Hu ◽

...

Keyword(s):

Sperm Quality ◽

Liquid Storage ◽

Boar Sperm

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Absence of seminal plasma from sperm-rich fraction decreases boar sperm quality characteristics during the course of liquid storage

Animal Reproduction Science ◽

10.1016/j.anireprosci.2018.08.029 ◽

2018 ◽

Vol 198 ◽

pp. 20-26 ◽

Cited By ~ 3

Author(s):

D.F. Leal ◽

M.A. Torres ◽

G.M. Ravagnani ◽

S.M.M.K. Martins ◽

F.V. Meirelles ◽

...

Keyword(s):

Seminal Plasma ◽

Sperm Quality ◽

Quality Characteristics ◽

Liquid Storage ◽

Boar Sperm

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In vitro effects of two different commercial freezing and thawing extenders on boar sperm quality

Animal Reproduction Science ◽

10.1016/j.anireprosci.2021.106906 ◽

2021 ◽

pp. 106906

Author(s):

De Andrade AFC ◽

R Grossfeld ◽

RV Knox

Keyword(s):

Sperm Quality ◽

Freezing And Thawing ◽

Boar Sperm ◽

In Vitro Effects

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Implication of Polyhistidine, a Novel Apoptosis Inhibitor, in Inhibiting Lipopolysaccharide-Induced Apoptosis in Boar Sperm

Animals ◽

10.3390/ani9100719 ◽

2019 ◽

Vol 9 (10) ◽

pp. 719 ◽

Cited By ~ 1

Author(s):

Tianzeng Song ◽

Yi Shi ◽

Yangang Wang ◽

Izhar Hyder Qazi ◽

Christiana Angel ◽

...

Keyword(s):

Sperm Quality ◽

Storage Conditions ◽

Potential Implication ◽

Liquid Storage ◽

Boar Sperm ◽

Tlr4 Agonist ◽

Reasonable Evidence ◽

Boar Semen ◽

Induced Apoptosis ◽

Apoptosis Inhibitor

Lipopolysaccharide (LPS) released from Gram-negative bacteria binds to toll-like receptor 4 (TLR4) and induces boar sperm apoptosis. Similarly, polyhistidine (pHis), a TLR4 agonist, can also bind to TLR4. We hypothesized that pHis could inhibit LPS-induced sperm apoptosis by competitively binding to TLR4 to then improve sperm quality. Therefore, the objective of this study was to examine whether pHis can inhibit LPS-induced sperm apoptosis and affect sperm quality. The results showed that the concentrations of bacterial colonies were significantly increased from 36 to 120 h under liquid storage conditions (p < 0.05); however, concentrations of LPS in boar semen showed a relatively constant trend (4.98 ± 1.55 EU/mL) following 120 h storage. The addition of 100 μg/mL pHis in the BTS extender significantly improved boar sperm motility and viability at 37 °C, and it significantly (p < 0.05) inhibited boar sperm apoptosis under liquid storage (17 °C) and at 37 °C incubation conditions. The co-treatment of LPS and pHis further confirmed that pHis played its role in inhibiting LPS-induced sperm apoptosis. In conclusion, our preliminary findings provide reasonable evidence that pHis could act as an inhibitor of LPS-induced apoptosis in boar sperm stored for longer periods of time. pHis might inhibit LPS-induced sperm apoptosis by competitively binding to TLR4. Nevertheless, further mechanistic studies are awaited to fully elucidate its potential implication in inhibiting LSP-induced apoptosis.

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Antioxidant Effect of a Polyphenol-Rich Murtilla (Ugni molinae Turcz.) Extract and Its Effect on the Regulation of Metabolism in Refrigerated Boar Sperm

Oxidative Medicine and Cellular Longevity ◽

10.1155/2019/2917513 ◽

2019 ◽

Vol 2019 ◽

pp. 1-15 ◽

Cited By ~ 3

Author(s):

Ignacio Jofré ◽

Magdalena Cuevas ◽

Leticia Signori de Castro ◽

João Diego de Agostini Losano ◽

Mariana Andrade Torres ◽

...

Keyword(s):

Lipid Peroxidation ◽

Sperm Motility ◽

Sperm Quality ◽

Membrane Damage ◽

Antioxidant Effect ◽

Boar Sperm ◽

Fertilization Capacity ◽

Ugni Molinae

The production of reactive oxygen species (ROS) in boar spermatozoa increases in refrigeration; this can have an impact on sperm quality and fertilization capacity. We evaluated the effect of polyphenol-rich aqueous extract of murtilla (Ugni molinae Turcz) on boar sperm stored at 17°C in order to reduce oxidative stress and improve sperm quality in the long term. Five experiments were performed: first, characterization of the polyphenol content from five genotypes of murtilla; second, determination of the genotype with the best antioxidant effect (MT-Ex); third, the antioxidant capacity on O2- and lipid peroxidation; fourth, the influence of MT-Ex on motility, calcium movement, cAMP, and metabolic parameters; and fifth, analysis of long-term refrigeration. The average phenolic content was 344 ppm; gallic acid, catechin, quercetin, myricetin, and kaempferol were detected. All extracts evaluated presented a concentration-dependent antioxidant effect. MT-Ex reduces intracellular O2-/peroxides but low lipid peroxidation. MT-Ex in nonstimulated ROS conditions reduces sperm motility, mitochondrial membrane potential, cAMP, and ATP, but the succinate dehydrogenase activity remained normal; also, we observed a reduction in calcium movement in in vitro sperm capacitation. The long-term analyses showed that MT-Ex improved sperm motility decay and reduced membrane damage and ROS at 168 h. Based on this study, we propose MT-Ex as a supplement in semen extenders.

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Effects of Isatis root polysaccharide on boar sperm quality during liquid storage and in vitro fertilization

Animal Reproduction Science ◽

10.1016/j.anireprosci.2019.106178 ◽

2019 ◽

Vol 210 ◽

pp. 106178 ◽

Cited By ~ 1

Author(s):

Zhiqiang Ren ◽

Weike Shaoyong ◽

Qian Li ◽

Lu Ma ◽

Junying Xiao ◽

...

Keyword(s):

In Vitro Fertilization ◽

Sperm Quality ◽

Vitro Fertilization ◽

Liquid Storage ◽

Boar Sperm

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Effect of N-Methylacetamide Concentration and Thawing Rate on Chicken Sperm Quality after Cryopreservation

Animals ◽

10.3390/ani10050824 ◽

2020 ◽

Vol 10 (5) ◽

pp. 824

Author(s):

Fabio Mosca ◽

Luisa Zaniboni ◽

Ahmad Abdel Sayed ◽

Nicolaia Iaffaldano ◽

Dominga Soglia ◽

...

Keyword(s):

Cell Membrane ◽

Kinetic Parameters ◽

Sperm Quality ◽

Membrane Integrity ◽

Motile Sperm ◽

Cell Membrane Integrity ◽

Before And After ◽

Freezing Procedure ◽

Thawing Rate

In seeking alternative cryoprotectants to glycerol for a reference chicken semen freezing procedure, the aim of the present study was to compare the effect of two concentrations of N-Methylacetamide (MA) and two thawing rates on the quality of frozen-thawed semen. Semen samples were diluted in Lake pre-freezing extender, including 0.1 M trehalose in presence of 6% or 9% MA, loaded into straws, frozen in nitrogen vapors, and stored in liquid nitrogen. The following thawing treatments were used: 5 °C for 100 s and 38 °C for 30 s. Sperm quality (cell membrane integrity, motility and kinetic parameters) was assessed before and after cryopreservation. The decrease of MA concentration from 9 to 6% improved sperm quality after freezing/thawing and this effect was dependent on thawing temperature. Decreasing the MA concentration from 9 to 6% improved the proportion of undamaged membrane, motile, and progressive motile sperm recovered after thawing at 5 °C for 100 s; in contrast, no effect of the MA concentration was observed thawing at 38 °C for 30 s. Therefore, the treatment with 6% MA and thawing at 5 °C for 100 s has given the best cryoprotective action. These results contribute to improve the efficacy of the current chicken semen cryopreservation procedures.

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