Search

Results of artificial insemination (AI) are affected by changes in sperm quality and the function throughout collection and preservation procedures. Proteome and metabolome alterations of sperm treated with the different procedures in goat, however, aren't fully understood. To this end, we sought to investigate the impacts of rectal probe electrostimulation (EE) and artificial vagina (AV) semen collection methods on the quality and the cryotolerance of goat sperm, with additional focus on proteomic and metabolomic analyses. Semen samples were collected from Yunshang black goats and categorized into four groups: fresh sperm collected via AV (XAZ), fresh sperm collected via EE (XEZ), frozen sperm post-AV collection (DAZ) and frozen sperm post-EE collection (DEZ). Four comparisons (XAZ vs. XEZ, DAZ vs. XAZ, DEZ vs. XEZ, DAZ vs. DEZ) were performed, respectively. This study first evaluated sperm motility, acrosome integrity, plasma membrane integrity, mitochondrial activity, and reactive oxygen species (ROS) levels. The results indicated that there were no significant differences in fresh sperm quality parameters between the EE and AV methods. However, notable differences emerged post-cryopreservation. Specifically, the AV method proved more advantageous in preserving the motility, integrities of acrosome and plasma membrane, mitochondrial activity of frozen sperm compared to the EE method. Through the multi-omics approaches, a total of 210 differentially abundant proteins (DAPs) related to sperm characteristics and function were identified across the four comparations. Moreover, 32 differentially abundant metabolites (DAMs) were detected. Comprehensive bioinformatics analysis underscored significant molecular pathways in the co-enrichment of DAPs and DAMs, particularly focusing on the citrate cycle, ROS, oxidative phosphorylation, and glycine, serine, and threonine metabolism etc. We elucidated the differential impacts of AV and EE collection methods on the quality and cryotolerance of goat semen from omics perspectives, which offer a critical foundation for further exploration into optimizing semen collection and cryopreservation techniques in goat breeding program., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

In sheep, artificial insemination serves as an important technique for breed improvement. In this context, genetic material from a small number of superior sires can be used in a large number of females. During this process, the storage of ram sperm may influence the efficiency of artificial insemination. Two main methods are currently used for ram semen storage: liquid storage and cryopreservation. The oxidative stress during the storage process can injure ram sperm and in some cases this leads to irreversible damage at the cellular level. To reduce such negative effects, different preservation protocols, extenders and protective components have been tested to improve ram sperm quality and to achieve greater fertility rates. This review provides an overview of the recent progress in extender supplementation using antioxidants and other compounds to improve ram semen quality parameters and fertility rates. It will emphasize on enzymes, vitamins, amino acids, proteins, some plant extracts and other compounds such as sugars, seminal plasma and fatty acids that can be used to supplement the extenders to reduce the formation of oxidants in ram semen and maintain its quality and enhance its fertility. It will also stress on how these supplements act, what were the tested levels giving beneficial effects on motility, viability, plasma membrane integrity and DNA fragmentation in liquid, cooled and post-thawing semen?, (Copyright © 2018 Elsevier B.V. All rights reserved.)

The present study aimed to assess the phenolic composition of the acetone extract from Opuntia ficus indica cladodes (ACTEX) and its effects on ram semen variables, lipid peroxidation and DNA fragmentation during liquid storage at 5°C for up to 72h in skim milk and Tris egg yolk extenders. Semen samples from five rams were pooled extended with Tris-egg yolk (TEY) or skim milk (SM) extenders containing ACTEX (0%, 1%, 2%, 4% and 8%) at a final concentration of 0.8×10 9 sperm/ml and stored for up to 72h at 5°C. The sperm variables were evaluated at different time periods (8, 24, 48 and 72h). Sperm total motility and viability were superior in TEY than in SM whereas the progressive motility, membrane integrity, abnormality and spontaneous lipid peroxidation were greater in SM compared to TEY (P<0.05). The results also indicated that the inclusion of 1% ACTEX in the SM or TEY extender increased the sperm motility, viability, membrane integrity, and decreased the abnormality, lipids peroxidation up to 72h in storage compared to control group. Similarly, even at 72h of storage, 1% ACTEX can efficiently decrease the negative effects of liquid storage on sperm DNA fragmentation (P<0.05). In conclusion, SM and TEY supplemented with 1% of ACTEX can improve the quality of ram semen. Further studies are required to identify the active components in ACTEX involved in its effect on ram sperm preservation., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Sperm cryopreservation is an approach to preserve sperm cells in liquid nitrogen or other cryogenic media for future use in assisted reproductive technologies, such as in vitro fertilization or artificial insemination. Sperm cryopreservation has been extensively used in the dairy industry and has attained excellent results after artificial insemination. However, for small ruminants the application of sperm cryopreservation is limited, due to the poor quality of frozen semen and special characteristics of the reproductive female tract. In order to improve post-thaw semen quality various cryoprotectants are used. Currently, many types of cryoprotectants, such as permeable organic solvents, sugars, antioxidants, and natural or synthetic ice blockers, have been tested on small ruminants' sperm cryopreservation. Among them, trehalose; has shown potential acting as an excellent cryoprotectant for semen freezing. While, the exact roles and action mechanisms of trehalose during cryopreservation remain unclear. In this review, we systematically summarized the present usage status, potential action mechanisms, and future application prospects of trehalose in small-ruminant sperm cryopreservation. [ABSTRACT FROM AUTHOR]

At present, there are few reports about the proteomics changes provoked by butylated hydroxytoluene (BHT) supplementation on cryopreserved semen in mammals. Thus, we aimed to evaluate the effects of different concentrations of BHT on goat sperm and to investigate the proteomics changes of adding BHT to cryopreserved goat (Capra hircus) sperm. Firstly, semen samples were collected from four goats, and frozen in the basic extenders containing different concentrations of BHT (0.5 mM, 1.0 mM, 2.0 mM) and a control without BHT, respectively. After thawing, the protective effects of dose-dependent replenished BHT to the freezing medium on post-thaw sperm motility, integrities of plasma membrane and acrosome, reactive oxygen species levels were confirmed, with 0.5 mM BHT being the best (B group) as compared to the control (without BHT, C group). Afterwards, TMT-based quantitative proteomic technique was performed to profile proteome of the goat sperm between C group and B group. Parallel reaction monitoring was used to confirm reliability of the data. Overall, 2,476 proteins were identified and quantified via this approach. Comparing the C and B groups directly (C vs. B), there were 17 differentially abundant proteins (DAPs) po-tentially associated with sperm characteristics and functions were identified, wherein three were upregulated and 14 were downregulated, respectively. GO annotation analysis demonstrated the potential involvement of the identified DAPs in metabolic process, multi-organism process, reproduction, reproductive process, and cellular process. KEGG enrichment analysis further indicated their potential roles in renin-angiotensin system and glutathione metabolism pathways. Together, this novel study clearly shows that BHT can effectively improve quality parameters and fertility potential of post-thawed goat sperm at the optimal concentration, and its cryoprotection may be realized through regulation of sperm metabolism and antioxidative capability from the perspective of sperm proteomic modification. [ABSTRACT FROM AUTHOR]

Objective: Recently, oral antioxidants in combined forms have been used to treat men with idiopathic infertility. This study aimed to evaluate the effects of treatment with vitamin C, vitamin E, selenium, zinc, arginine, L-carnitine, and coenzyme Q10 on sperm quality parameters, DNA integrity, reproductive hormones, and pregnancy rates in men with infertility and idiopathic oligoasthenoteratozoospermia (OAT). Methods: A prospective study was conducted on 420 men with infertility and idiopathic OAT who took an oral supplement of antioxidant SP-Power tablets twice daily for 6 months. Semen quality, reproductive hormones, and the DNA fragmentation index (DFI) were evaluated at baseline and at 3 and 6 months after supplementation, using the World Health Organization 2021 guidelines. Results: No significant difference was observed in volume or the percentage of typical morphology during treatment. A significant improvement in sperm concentration was observed after supplementation (8.67±1.41, 12.17±1.91, and 19.01±0.86 at baseline, 3, and 6 months respectively, p<0.01). The total motility, progressive motility, and total motile sperm count also increased significantly (p<0.01), whereas the DFI decreased after 6 months. There was an increase in normal FSH levels and testosterone levels after 6 months of supplementation of antioxidant SP-Power, but these differences were not statistically significant (p=not significant and p=0.06, respectively). Conclusion: Supplementation with SP-Power tablets improved sperm quality parameters, sperm DFI, some reproductive hormones, and pregnancy rates in men with infertility and idiopathic OAT, which could be attributed to the supplement's synergistic antioxidant action. Further studies are needed to determine the effects of supplementation on oxidative stress markers. [ABSTRACT FROM AUTHOR]

Background: DNA fragmentation index (DFI) enhances routine semen analysis by providing valuable insights into male reproductive potential. Utilizing Halosperm test, a sperm chromatin dispersion (SCD) assay based on induced condensation. The purpose of this study was to assess sperm DNA damage both before and after freezing. By following the specified kit instructions, an attempt was made to validate the SCD test protocol, with a particular emphasis on the implications of sperm freezing on its DNA integrity. Methods: In total, 380 fresh human semen samples from normozoospermic patients were frozen at -20°C for 10 days, using SCD cryopreservation reagent. Routine semen analysis and DNA fragmentation index (DFI) were determined for each sample before freezing and after thawing. Semen morphology and sperm DFI were compared before and after freezing/thawing process. Results: There was a significant decrease in sperm normal morphology after thawing (9.31±2.42% vs. 7.1±1.53%, p<0.05, respectively). The sperm head, midpiece, and tail defect rate increased after freezing at -20°C. Moreover, DFI was significantly higher after thawing compared to before freezing (20.71±1.61% before freezing vs. 29.1±0.21% after thawing with p<0.001). Conclusion: Cryoconservation of semen samples at -20°C for 10 days using SCD cryopreservation reagent seems to damage sperm morphology, resulting in a reduction in sperm DNA integrity. The measurement of DFI on a fresh sample remains the most reliable technique for obtaining accurate results. [ABSTRACT FROM AUTHOR]

Recently, researchers have focused on the use of natural antioxidants to improve semen quality as a key element for successful artificial insemination. In this context, the first aim of this study was to determine the antioxidant activity and composition (minerals, vitamins, and sugars) of Opuntia ficus-indica cladode ethanolic extract (ETHEX). A further purpose of the study was to investigate the effect of ETHEX supplementation on the quality of liquid ram semen extended with skim milk (SM) at 5°C. The antioxidant activity of ETHEX was studied using free radical 1, 1-diphenyl-2-picrylhydrazyl (DPPH•) assay. The mineral composition and the sugar and vitamin contents of ETHEX were determined using an inductively coupled plasma optical emission spectrometry (ICP-OES) and HPLC-DAD-RID analytical instruments. As a second part, semen was collected from five Boujaâd rams with an artificial vagina. The ejaculates with more than 70% motility were pooled, extended with skim milk (SM) extender without (control) or supplemented with 1–8% of ETHEX (37°C; 0.8 × 109 sperm/mL). Sperm quality parameters were assessed at 8, 24, 48, and 72 h. The results showed that ETHEX had a higher antioxidant activity compared to those of ascorbic acid and butylated hydroxytoluene (BHT). Furthermore, ETHEX contains a considerable amount of minerals, vitamins, and sugars. The inclusion of 1 or 2% ETHEX in SM increased the sperm motility, viability, and membrane integrity and decreased the abnormality of spontaneous and catalyzed lipids peroxidation (p < 0.05) up to 72 h. In addition, semen diluted with 1 and 2% ETHEX decreased the level of DNA fragmentation compared to the control group (p < 0.05). In conclusion, the ETHEX could be recommended to improve the quality of liquid ram spermatozoa. However, its effects on artificial insemination should be further studied. [ABSTRACT FROM AUTHOR]

The aim of this study was to see how chilled sperm quality is affected by ejaculation frequency and its correlation to seminal plasma composition in INRA 180 rams. Five rams were collected at high (HFE) and low frequencies (LFE). For the high frequency, the rams were collected three times on the same day every three days for 18 days. In the low frequency collection, three consecutive ejaculates were collected once a week for four weeks. Ejaculates were collected at 20-minute intervals in either HFE or LFE. Semen characteristics, concentration of total protein, lipid, cholesterol and fructose in seminal plasma were assessed. Semen samples were extended in skim milk-based extender at 15 °C, then evaluated at different storage times (0, 8, and 24 h). Fresh sperm quality parameters, seminal plasma composition, and stored sperm quality were shown to be higher in LFE than in HFE and in the first and second ejaculates than in the third one. After 24 h of storage, sperm quality was correlated to seminal plasma components. In conclusion, the frequency of ejaculation has an effect on the fresh and stored semen quality as well as on seminal plasma composition in INRA 180 rams. [ABSTRACT FROM AUTHOR]

The purpose of this investigation was to evaluate the effect of cactus seed oil (CSO) on ram semen quality parameters during liquid storage. Semen samples collected from five adult fertile Boujaâd rams were pooled and diluted to a final concentration of 0.8 × 10 9 spermatozoa/ml in Tris egg yolk (TEY) and skim milk (SM) extenders supplemented with CSO at 0, 1, 2, 5, and 10% v/v. The samples were then stored at 5 °C and several sperm parameters (motility, viability, morphology, peroxidation and DNA fragmentation) were assessed after 8, 24, 48 and 72 h. Results showed that the supplementation of TEY and SM extenders with 1% and 2% CSO, respectively, increased sperm motility and viability and decreased the level of peroxidation and DNA fragmentation during storage. Therefore, we concluded that 1% and 2% CSO are the optimal concentrations that can help to improve ram semen quality during storage at 5 °C. [ABSTRACT FROM AUTHOR]

Due to its high antioxidant content, the argan oil could play a beneficial role in liquid storage of ram semen. The aim of this study was to investigate effects of different concentration of argan oil (ARO) on spermatologic parameters, lipid peroxidation and DNA fragmentation during liquid storage of ram semen until 48 h. Also effects of extenders and temperature on same parameters were assessed. For these aims, semen samples were collected from Boujaâd rams, extended with Tris egg yolk or skim milk extenders without (control) or supplemented with different concentrations of ARO (1%, 2%, 5% and 10% v/v) at a final concentration of 0.8 × 10 9 sperm/mL and stored until 48 h at 5 °C or 15 °C. The sperm quality assessments were performed at different intervals during storage (0, 8, 24 and 48 h). Sperm progressive motility started to decrease after 8 h of storage in all temperatures – extenders combinations and dropped steadily during the 8–48 h interval. However, sperm viability, progressive motility and membrane integrity were markedly higher in ARO groups (especially in 1% in Tris and 5% in skim milk) until 24 h and 48 h storage at both temperatures compared to controls. The argan oil also decreased the level of spontaneous and induced malondialdehyde (MDA) and the sperm DNA fragmentation until 48 h storage. In conclusion, it was determined that addition of argan oil to conventional extenders may improve the quality of ram semen during liquid storage in different temperatures. [ABSTRACT FROM AUTHOR]

Highlights • In Boujaâd ram, the semen quality parameters, were affected by age and season. • In Boujaâd ram, the semen liquid storage was affected by age and season. • The advanced age negatively affects quality and storage of semen in Boujaâd rams. • Motility of stored semen was highly correlated to seminal plasma composition. Abstract The present research was conducted to evaluate the effects of season and age on scrotal circumference (SC), semen characteristics, semen composition, and sperm motility during liquid storage in Boujaâd rams. Semen was collected throughout 1 year from April 2014 to March 2015. Two ram groups, namely older rams including animals aged 5.5 to 6 years of age, and younger rams including animals aged 2.5 to 3 years of age were used. Scrotal circumferences, semen characteristics, and some biochemical variables from semen were measured. After collection and evaluation, the semen was extended in skim milk (SM) at 15 °C to a concentration of 0.8 × 109 spermatozoa/ml. Thereafter, samples were evaluated at different storage times (0, 8, and 24 h). The results indicate that older rams had greater values of SC, lesser semen quality, protein, lipid, and cholesterol concentrations than younger rams. Nevertheless, for both groups there were the least values for semen quality and semen composition in winter. The semen collected from younger rams had greater motility after liquid storage compared with that of the older rams. The least semen quality after liquid storage was recorded in the winter compared with the other seasons of the year for both age groups. In conclusion, results of the present study indicate that older Boujaâd sheep have lesser semen quality, different semen composition than younger rams after liquid storage and during different seasons of the year. Furthermore, there was a strong correlation between semen composition and motility of stored ram semen. [ABSTRACT FROM AUTHOR]

The present study was undertaken to assess the effect of seasons on scrotal circumference, semen characteristics, seminal plasma composition, and sperm motility during liquid storage of INRA180 rams. The semen was collected from five mature INRA180 rams (2–3 years of age) during one year (from April 2014 to March 2015). Scrotal circumferences, semen characteristics, some biochemical parameters of seminal plasma were evaluated. Immediately after collection and evaluation, the semen was pooled and extended in skim milk (SM) at 15 °C to reach 0.8 × 109 spermatozoa/ml. Thereafter, samples were evaluated at different storage times (0, 8, and 24 h). The results showed that scrotal circumference, semen quality and the concentration of total protein in seminal plasma were relatively constant during the year ( P > 0.05). However, total lipid and cholesterol concentrations increased significantly ( P < 0.001) in winter and summer. The result showed also that progressive motility was higher in winter and summer after 24h of storage ( P < 0.01). In contrast, no difference was recorded regarding total motility ( P > 0.05). To conclude, the INRA180 rams have the ability to produce semen with high quality all over the year. The only parameters showing seasonal variations are cholesterol, total lipid, and progressive motility. [ABSTRACT FROM AUTHOR]