Higher Sperm Motility Research Articles

Selenium efficiency in protecting sperm quality and testicular parameters of South African indigenous Zulu rams exposed to heat stress.

The study investigated selenium's (Se) efficiency in preserving South African Zulu rams' sperm quality and testicular parameters when they were exposed to heat stress. Indigenous Zulu rams (20) between 2 and 5 years old were allocated into four groups, namely the Se, testicular heat stress (THS), selenium plus testicular heat stress (SeTHS), and control. Each group comprised five rams; the groups were balanced according to the rams' body weight and scrotal circumference. The Se and SeTHS groups received sodium selenite orally bi-weekly for 5 months. To induce heat stress, testicular heat insulation bags were wrapped around the testes of the rams receiving the THS and SeTHS treatments for 49 days. Semen was collected from the rams weekly from the third month onward; the first two months were for Se & THS acclimatization. In addition, testicular measurements were taken bi-weekly. Analysis of variance was used to analyze the sperm quality data. Duncan's multiple range test was used to compare the groups' data for significant differences. The results showed that the Se-supplemented rams' scrotal circumference was smaller (p < 0.05) compared with the other groups. The Se, SeTHS, and control groups demonstrated similar total sperm motility; in contrast, the THS and SeTHS groups recorded low and high total sperm motility, respectively, compared with other treatment groups (p < 0.05). The semen from the rams that received THS without Se displayed a significantly higher number of immotile sperm cells (p < 0.05) and poor sperm quality, including total and progressive motility, and kinematic parameters when compared with other treatments, suggesting that Se protects sperm against THS. We concluded that selenium protected some sperm parameters (TSM, PSM, MV, VCL, VSL) of THS- treated rams while others did not improve (RV, NSM, C, STR).

Proteomics and Metabolic Characteristics of Boar Seminal Plasma Extracellular Vesicles Reveal Biomarker Candidates Related to Sperm Motility.

Although seminal plasma extracellular vesicles (SPEVs) play important roles in sperm function, little is known about their metabolite compositions and roles in sperm motility. Here, we performed metabolomics and proteomics analysis of boar SPEVs with high or low sperm motility to investigate specific biomarkers affecting sperm motility. In total, 140 proteins and 32 metabolites were obtained through differentially expressed analysis and weighted gene coexpression network analysis (WGCNA). Seven differentially expressed proteins (DEPs) (ADIRF, EPS8L1, PRCP, CD81, PTPRD, CSK, LOC100736569) and six differentially expressed metabolites (DEMs) (adenosine, beclomethasone, 1,2-benzenedicarboxylic acid, urea, 1-methyl-l-histidine, and palmitic acid) were also identified in WGCNA significant modules. Joint pathway analysis revealed that three DEPs (GART, ADCY7, and NTPCR) and two DEMs (urea and adenosine) were involved in purine metabolism. Our results suggested that there was significant correlation between proteins and metabolites, such as IL4I1 and urea (r = 0.86). Furthermore, we detected the expression level of GART, ADCY7, and CDC42 in sperm of two groups, which further verified the experimental results. This study revealed that several proteins and metabolites in SPEVs play important roles in sperm motility. Our results offered new insights into the complex mechanism of sperm motility and identified potential biomarkers for male reproductive diseases.

Lipidomic and transcriptomic characteristics of boar seminal plasma extracellular vesicles associated with sperm motility

Seminal plasma extracellular vesicles (SPEVs) play an important role in regulating sperm motility by delivering various cargoes, such as miRNAs, mRNAs, proteins and metabolites. However, information on the lipid compositions of SPEVs and their roles in semen quality is limited. Here, we performed high-throughput transcriptomic and lipidomic analysis on SPEVs isolated from 20 boars with high or low sperm motility. Then, we evaluated the lipid composition and gene expression characteristics of SPEVs and identified the specific lipids and genes related to sperm motility. As a result, a total of 26 lipid classes were identified in SPEVs, and five subclasses, CerG2, CerG3, LPE, LPS and TG, were significantly different in boars with high and low sperm motility. In addition, 195 important lipids and 334 important genes were identified by weighted gene coexpression analysis (WGCNA) and differential expression analysis. We observed that several important genes and lipids in SPEVs potentially influence sperm motility via glycerophospholipid metabolism, glycerolipid metabolism, the sphingolipid signaling pathway and the ferroptosis pathway. Furthermore, we found a significant correlation between the content of 22 lipids and the expression levels of 67 genes (|cor| > 0.8, P < 0.05). Moreover, we observed that three important gene-lipid linkages (CerG1 (d22:0/24:0) - RCAN3, Cer (d18:1/24:0) - SCFD2 and CerG1 (d18:0/24:1) - SCFD2) were strongly correlated with sperm motility. Based on the results, some genes and lipids in SPEVs may play important roles in sperm motility by interacting with sperm through important pathways.

Seminal plasma removal for medium-term preservation of ram sperm at 5 °C

This study aimed to investigate if washing ram sperm from seminal plasma (SP) could be an effective tool to extend sperm lifespan in medium-term preservation in liquid form to optimize ovine artificial insemination protocols. To this end, in Experiment 1 SP was added to a sperm model without previous contact with this substance (ram epididymal sperm) at the beginning or the end of a 48-hour preservation protocol at 5 °C (n = 13). Sperm motility and kinetic parameters and sperm functionality in terms of sperm viability, apoptosis, mitochondrial activity and reacted acrosomes were assessed after 6 h of storage at 15 °C (standard liquid preservation method) and 24 and 48 h at 5 °C. Extended sperm showed better results after 48 h when stored in the absence than in the presence of SP in most sperm quality parameters. Moreover, the final SP supplementation of this experimental group resulted in the highest sperm motility and kinetic parameters, viability and mitochondrial activity. These results suggested that initial SP deprivation could be beneficial in a medium-term ram sperm preservation protocol in liquid form, as well as a final supplementation. Therefore, we conducted Experiment 2 to evaluate the effect of SP removal from freshly ejaculated ram semen under the same storage conditions as in Experiment 1 (n = 12). Surprisingly, SP withdrawal impaired sperm functionality, leading to increased apoptosis and decreased mitochondrial activity after 24 and 48 h at 5 °C. Conversely, SP supplementation at the end of the preservation protocol of the ejaculate processed as usual had a positive effect on sperm quality and fertility. To summarize, SP absence was beneficial for a medium-term preservation protocol (up to 48 h at 5 °C) of ram epididymal sperm, but the same preservation protocol for ram ejaculated sperm revealed a possible failure of the SP removal method in avoiding the sperm-SP interaction effect. Meanwhile, SP supplementation of ram semen at the end of the preservation protocol increased in vitro sperm quality and fertility after artificial insemination.

Key miRNAs of chicken seminal plasma extracellular vesicles related with sperm motility regulation

MicroRNAs (miRNAs) are bio-active elements cargoed by seminal plasma extracellular vesicles extracellular vesicles (SPEVs) which are crucial for sperm function and fertility modulation. This study aimed to isolate, characterize, and identify the miRNA expression profiles in the SPEVs from high (HSM) and low sperm motility (LSM) groups that could serve as fertility biomarkers and explain the underlying mechanisms. The isolated SPEVs were round spherical structures of approximately 50–200 nm in diameter expressing molecular markers. A total of 1006 and 1084 miRNAs were detected in HSM and LSM, respectively, with 34 being differentially expressed. Their targeted genes involved in SNARE interactions in vesicular transport, Metabolic pathways, and Apelin signaling pathway, etc. The joint analysis with mRNAs of sperm and sperm storage tubules cells highlighted the cellular communication mediated by SPEVs miRNAs, where they may rule fertility by affecting sperm maturation and amino acid metabolism. SPEVs as additives could improve fertility of fresh and frozen sperm, while the knockdown of one of the differentially expressed miRNAs, miR-24-3p, diminished this effect, indicating its crucial roles. This study expands our understanding of SPEVs miRNAs mediated sperm maturation and fertility modulation, and may help to develop new therapeutic strategies for infertility and sperm storage.

Proteomic profile of seminal plasma from Pêga donkeys (Equus asinus) with high sperm motility and vigor: Implications for assisted reproduction

Donkeys of the Pêga breed (Equus asinus) have been used for two centuries to produce breeding stock and create hybrids for labor and transport in southeast Brazil, and for exporting meat and milk to other countries. Furthermore, they are used in competitions, as they are docile and easy to handle. However, assisted reproduction success rates for frozen donkey semen are remarkably low, with no standardized method for cryopreserving sperm after removal of seminal plasma. This work aims to reveal the biological involvement of seminal plasma proteins from Pêga donkeys in aiding the development of assisted reproduction. This study was carried out with 14 ejaculates collected every eight days, throughout the breeding season, from three healthy fertile Pêga donkeys, with an average age of four years. After confirming the high freezability of fresh semen by evaluating quality parameters, the seminal plasma was separated by centrifugation and an aliquot from each collection was microfiltered and frozen. A label-free technique followed by LC-MS/MS analysis applied to pools of seminal plasma samples from each animal revealed 522 proteins in the proteomic profile, of which 49.8 % (260 proteins) are related to cellular energy transformation, and many proteins involved in reproduction (76), spermatogenesis (38), fertilization (29), among other biological process. By comparison with literature, Pêga donkeys share many proteins with donkeys of Dezhou breed that present great potential as fertility biomarkers. Our results showed proteins positively related to fertilization for different breeds of donkeys around the world, helping to enhance the assisted reproduction of Pêga donkeys.

Effect of different concentrations of bovine serum albumin on freezing of goat semen

Objective. Evaluate the effect of Bovine Serum Albumin (BSA) concentration on the freezability of goat semen in a Tris-based lyophilized preservative-diluter, without performing seminal washing, compared with a control preservative-diluter of lactose-skimmed milk (DC), removing the seminal plasma by centrifugation. Materials and methods. It will cover 90 ejaculates, volume, motility, concentration, viability, and sperm morphology. The fit ejaculates were mixed and the pool divided into five portions, each receiving one of 4 lyophilized combinations based on Tris-Glucose-Ac. Citrus and Glycerol with different concentrations of BSA (0.1%, 0.5%, 1% and 2%) or DC. It was frozen in 0.1 ml pellets in nitrogen vapors, and after 2 min., They were stored in liquid nitrogen until thawing 7 days later, the following were determined: motility percentage (30, 120 and 240 minutes), viability, damaged acrosomes and total anomalies (30 and 120 minutes) and were compared using a Binary Logistic Regression model. Results. The highest sperm motility and viability (p &lt;0.05) in the three times was for 0.5%, 1% and 2% of BSA, which were higher than 0.1% of BSA and DC. Damages acrosome and total abnormalities at 30 and 120 minutes were lower (p &lt;0.05) for 0.5%, 1% and 2% of BSA compared to 0.1% and DC. Conclusion. The cryopreservation of goat semen does not require seminal washing by centrifugation if 0.5-2% BSA is used as a membrane protector in a lyophilized dilute-conservative based on Tris-Glucose-Citric Acid and Glycerol.

The effect of LINC9137 targeting miR-140-3p-NKAIN3 signal axis on the development of goose testis sertoli cells

Sertoli cells (SC) are a type of important cells in the testes, which can provide transport proteins, regulatory proteins, growth factors, and other cytokines for the spermatogenic process. They participate in the regulation of the maturation and differentiation of spermatogenic cells and play an important supporting role in the migration, proliferation, and differentiation of germ cells at all levels in the testes. Previous studies found differential expression of LINC9137, miR-140-3p, and Sodium/Potassium Transporting ATPase Interacting 3 (NKAIN3) genesin high and low sperm motility goose testicular tissues. This study investigated the effects of the LINC9137-miR-140-3p-NKAIN3 signal axis on the proliferation and apoptosis of goose testicular sertoli cells at the cellular level, respectively. The results showed that through acridine orange staining, oil red O staining, Alkaline phosphatase (AKP) staining, and RT qPCR assay, it was comprehensively identified that the cultured testicular sertoli cells were purified in vitro. Through the dual luciferase activity detection test, it was found that LINC9137 has a targeted binding site with miR-140-3p and NKAIN3. In addition, this study found that overexpression of miR-140-3p significantly inhibited the expression of LINC9137 and NKAIN3 in sertoli cells, and their expression was significantly increased when miR-140-3p was interfered with. By measuring cell proliferation activity and apoptosis related gene expression, it was found that overexpression of LINC9137 decreased cell proliferation activity (P > 0.05), while the expression level of apoptosis factor Bcl2 Associated X Protein (Bax)/B-cell lymphoma-2 (Bcl2) increased (P > 0.05). On the contrary, when interfering with LINC9137, the cell proliferation activity of sertoli cells was significantly increased (P < 0.01), and the expression level of apoptosis factor Bax/Bcl2 was significantly reduced (P < 0.05); The effect of miR-140-3p on the proliferation and apoptosis of sertoli cells is opposite to that of LINC9137. Meanwhile, this study co transfected overexpressed LINC9137 and miR-140-3p plasmids into sertoli cells, and found that the effect of LINC9137 overexpression on supporting cell proliferation was weakened by miR-140-3p. This study elucidates the role and function of the LINC9137 miR-140-3p-NKAIN3 signaling axis in the development of goose testes and spermatogenesis, establishes a regulatory network related to spermatogenesis, and provides a theoretical basis for studying the genetic regulation of goose spermatogenesis.

Evaluation of the effects of glucose concentration in extender and sperm concentration in straw for cryopreservation of Atlantic sturgeon (Acipenser oxyrinchus) semen

The Atlantic sturgeon (Acipenser oxyrinchus) faces unique conservation challenges due to its extended maturation period and irregular spawning patterns, which restrict its reproductive capacity. Therefore, in order to overcome these difficulties we have made a first attempt to develop effective cryopreservation procedure for Atlantic sturgeon through evaluation of the effects of the glucose concentration in extender and final sperm concentration in straw on sperm motility parameters. A remarkably high (81 ± 8%) postthaw sperm motility was obtained. The other sperm velocity and trajectory parameters were differentially affected both by concentrations of glucose or sperm and cryopreservation process. Our results undoubtedly show that the post-thaw sperm motility of Atlantic sturgeon is highly influenced by the final glucose and sperm concentration. The recommended cryopreservation procedure includes 0.05 M glucose and 7.5% methanol at 1.0×109 spermatozoa ml−1. We have confirmed that our elaborated protocol can be used for cryopreservation of Atlantic sturgeon semen which can be subsequently used for long-term conservation of this endangered species, offering a valuable tool in efforts to safeguard its genetic diversity and contribute to its survival.

Insights into alternative cryoprotectants to freeze sperm of domestic cats.

Globalization and habitat destruction pose a significant threat to wildlife felids. Even though conservation banks for genetic materials have been created, the sperm cryopreservation with minimal cell damage is still a great challenge. Thus, this study aimed to compare the effects of two commercial extenders with different concentrations of alternative cryoprotectants on thawed sperm quality of domestic cats. Five adult cats were anaesthetized (using a combination of 40 μg/kg medetomidine associated to 5 mg/kg ketamine), and the semen was collected by electroejaculation (electrical stimulation of 2-3 V). Semen samples were evaluated for sperm characteristics (kinetics, morphology, membrane integrity and morphometry). Subsequently, they were sorted into two aliquots and centrifuged. The aliquots were added to a commercial extender containing 3% glycerol and 2% methylformamide (extender I) or 2% glycerol and 3% methylformamide (extender II), frozen, thawed (37°C/30 s) and reevaluated. Comparatively, the sperm kinetics and membrane integrity of fresh semen were higher (p < .002) than frozen samples in extender I and II. Total and progressive motility were lowest in the thawed samples. However, the subjective analysis indicated high sperm motility, since the kinetics evaluation was impaired by the low cell number in the thawed samples. There were no differences in sperm morphology between the groups. In the sperm morphometric analysis, a significant difference (p = .04) was identified in the length of the intermediate piece in extender II samples compared with fresh and extender I. Thus, it can be concluded that although the concentrations tested did not maintain the kinetic parameters and membrane integrity of spermatozoa after thawing, the extender with a lower concentration of glycerol was less toxic for maintaining the midpiece length.

A retrospective analysis investigating the effects of Telazol® and medetomidine on ejaculate characteristics in cheetahs (Acinonyx jubatus)

Zoo managed cheetahs provide an insurance population for wild cheetahs that are under threat of extinction from habitat loss, lack of prey, competition, pet trade and poaching for skin and bones. Assisted reproductive techniques including artificial insemination, in vitro fertilization, and embryo transfer augment natural breeding programs but rely on good quality semen for best results. It is understood that anesthesia can affect semen characteristics such as ejaculate volume, total sperm count, sperm motility, and incidence of urine contamination. Thus, the aim of this study was to conduct a retrospective analysis of 23 years of data to investigate sperm parameters of semen collected under anesthesia using medetomidine in combination with butorphanol and midazolam or Telazol® alone. Electroejaculation records (Medetomidine, Butorphanol, and Midazolam anesthetized n = 59 ejaculates, from 30 cheetahs, Telazol® anesthetized, n= 169 ejaculates, from 72 cheetahs) were evaluated for incidence of urine contamination. Electroejaculation records (Medetomidine, Butorphanol, and Midazolam anesthetized n = 21 ejaculates, from 17 cheetahs, Telazol® anesthetized, n = 143 ejaculates, from 63 cheetahs) were evaluated for total sperm count, total motility, ejaculate volume, and testicle size. Telazol® treated cheetahs had a numerically higher total sperm count (Median ± SD: 42.58 ± 77.8 × 106 spermatozoa) compared to those treated with medetomidine (Median ± SD: 31.2 ±44.58 × 106 spermatozoa), and a significantly (p &amp;lt; 0.05) higher sperm motility (Median ± SD: 70.0 ± 9.71%) compared to medetomidine (Median ± SD: 53.0 ± 16.41%) treated cheetahs. The findings of this study indicate that medetomidine anesthesia results in significantly lower sperm motility and Telazol® anesthesia results in a higher total sperm count and motility, thus resulting in higher quality ejaculate. This information can aid in the veterinary management of the species when involved in genome resource banking and assisted reproductive technologies.

Oxidative phosphorylation rather than glycolysis is the primary energy source for sperm motility in the mussels Mytilus edulis

Broadcast-spawning marine mussels rely on high sperm motility for successful fertilization in the dynamic seawater environment. Mitochondria are typically considered the primary source of ATP generation via oxidative phosphorylation (OXPHOS); however, the ATP generation pathways of mussel sperm have not been fully characterized. To better understand the importance of both OXPHOS and glycolysis for mussel sperm function, we conducted experiments inhibiting these pathways in sperm from Mytilus edulis. Our results indicate that oligomycin, an inhibitor of the mitochondrial ATP synthase, immediately decreased sperm motility rate, velocity, and ATP content, while 2-deoxy-d-glucose, a glycolysis inhibitor, had no effect. The OXPHOS inhibitor rotenone also partially reduced sperm motility rate and velocity. Interestingly, no evidence was found for the inhibitors' effects on the content of energy-rich compounds (lipids, carbohydrates, and proteins) in the mussels' sperm, indicating only modest energy demand to fuel sperm motility. Based on these findings, we conclude that OXPHOS is the primary energy source for sperm motility in marine mussels. Our study sheds light on the intricacies of mussel sperm physiology and highlights the importance of understanding the energy requirements for successful fertilization in broadcast-spawning marine invertebrates.

Association between Yili goose sperm motility and expression profiles of mRNA and miRNA in testis

BackgroundThe study was conducted to find out the candidate microRNA (miRNA) and genes that associated with sperm motility of Yili goose through small RNA sequencing of testicular tissue of Yili goose, and provide a theoretical basis for the study of the regulation mechanism of sperm motility of Yili goose gander.ResultsIn this study, five male geese with high sperm motility and five male geese with low sperm motility were slaughtered to obtain their testis tissues for small RNA sequencing, and biological information methods were used for data analysis. The results showed that a total of 1575 known miRNAs and 68 novel miRNAs were identified in the testis tissue of Yili goose, and 71 differentially expressed miRNAs and 660 differentially expressed genes were screened. GO functional analysis showed that miRNAs target genes were mainly involved in the binding, kinase activity, structural constituent of cytoskeleton and intermediate filament cytoskeleton. KEGG functional analysis showed that miRNAs target genes were significantly enriched in arginine and proline metabolism, glycolysis / gluconeogenesis, fructose and mannose metabolism and beta-Alanine metabolism and other pathways. miRNAs-mRNAs interaction network suggests miR-140/miR-140-3p/miR-140-3p-NKAIN3, let-7d-BTG1 and miR-145-5p/miR -145a-5p-CLEC2E may play an important role in testis development and spermatogenesis.ConclusionsThe results of this study suggest that the sperm motility of Yili goose may be regulated by different miRNAs, and the target genes are significantly enriched in pathways related to sperm metabolism, indicating that miRNAs affect the sperm motility of Yili goose by regulating the metabolic process of sperm and the expression of related genes. This study can provide a reference for revealing the regulation mechanism of Yili goose sperm motility at the molecular level.

Identification of key genes affecting sperm motility in chicken based on whole-transcriptome sequencing

Sperm motility is an important index for the evaluation of semen quality. Improving sperm motility is important to improve reproductive performance, promote breeding process, and reduce production cost. However, the molecular mechanisms regulating sperm motility in chickens remain unclear. In this study, histological observation and whole-transcriptome analysis were performed on testicular tissue of chickens with high and low sperm motility. Histological observations showed that roosters with high sperm motility exhibited better semen quality than those with low sperm motility. In addition, the germinal epithelial cells of roosters with low sperm motility were loosely arranged and contained many vacuoles. RNA-seq results revealed the expression of 23,033 mRNAs, 2,893 lncRNAs, and 515 miRNAs in chicken testes. Among them, there were 417 differentially expressed mRNAs (DEmRNAs), 106 differentially expressed lncRNAs (DElncRNAs), and 15 differentially expressed miRNAs (DEmiRNAs) between high and low sperm motility testes. These differentially expressed genes were involved in the G protein-coupled receptor signaling pathway, cilia structure, Wnt signaling, MAPK signaling, GnRH signaling, and mTOR signaling. By integrating the competitive relationships between DEmRNAs, DElncRNAs, and DEmiRNAs, we identified the regulatory pathway of MSTRG.3077.3/MSTRG.9085.1-gga-miR-138-5p-CADM1 and MSTRG.2290.1-gga-miR-142-3p-GNAQ/PPP3CA as crucial in the modulation of chicken sperm motility. This study provides new insights into the function and mechanism of ceRNAs in regulating sperm motility in chicken testes.

Sperm quality of Bali bull following sexing and freezing using different cryoprotectants

The objective of this experiment was to assess the impact of sperm sexing duration on sperm quality and to compare the effectiveness of glycerol (GC) and ethylene glycol (EG) as cryoprotectants to preserve the quality of sexed sperm during the freezing and storage in liquid nitrogen. The treatments consisted of three different durations of sexing: 40 min.(D-40), 50 min (D-50), 60 min. (D-60). The variable measured was the motility and viability of sperm following sexing and post-thawing motility of Bali bull sexed sperm cryopreserved in GC and EG. The results showed that sperm motility after sexing was significantly higher in the treatment of 40 and 50 min. compared to 60 min. of sexing duration. While the viability of sexed sperm in all treatments was not significantly different. Post-thawing motility of frozen sexed sperm cryopreserved with GS was higher than sexed sperm cryopreserved with EG. Finally, it was concluded that sexing times of 40 and 50 min. were the best time to get high sperm motility and viability of Bali bull compared to sexing times of 60 min. In addition, GS could maintain the motility of sexed sperm better than EG.

Manipulated sperm motility via soot nanoparticles-induced biochemical alterations in human seminal plasma

Obtaining spermatozoa with progressive motility, via postejaculatory activation with pharmacological agents such as theophylline and pentoxifylline, is crucial for the success rate of assisted reproduction in couples with severe male factor infertility. Regrettably, the possibility of premature acrosome reactions and impared oocyte function questions the practical applicability of phosphodiesterase inhibitors. The rapid development of nanotechnologies promotes the use of hydrophobic rapeseed oil soot as a non-cytotoxic biomaterial for sperm motility activation, but the scarcity of knowledge regarding the interactions of soot with components from the seminal plasma hinders the eventual commercialization of this cutting-edge approach. Aiming to eliminate this shortcoming, the current study shows for the first time how the soot nanomaterials alter the biochemistry of human seminal plasma. Upon 270 min incubation with soot nanoparticles, the activity of AST, ALT, CK, LDH and GGT enzymes in the seminal plasma of ten patients changes inversely to the registered sperm motility (i.e., lower enzyme activity, higher sperm motility and vice versa). This phenomenon is primarily related to termination of the enzymes-substrate binding or extraction of enzymes from the gametes via chemical bonding with the soot. These novel mechanisms depend on the physicochemical features of used carbon nanomaterials, revealing opportunities for predictable tuning of the sperm reproductive potential.

Exploration of The Natural Extender for Dilution of Walking Catfish Clarias batrachus Sperm in Refrigerated Storage

Walking catfish Clarias batrachus is a freshwaterr fish that has high economic value. Currently, the culture of this local catfish has been initiated in Indonesia, but the breeding technology has not been well developed. Therefore, this study aims to explore the type of natural extender for sperm preservation of local catfish C. batrachus so that it can support the development of its breeding technology, especially in remote areas. The research was conducted at the Hatchery Laboratory of the Faculty of Marine and Fisheries, Universitas Syiahh Kuala, Banda Aceh, Indonesia.. Four types of natural extenders were tested in this study, namely young coconut water, nira water, sugarcane water, and old coconut water. Completely randomized design method consisting of five treatments and three replications were used in this study. Sperm were diluted with each tested extender with a sperm dilution rate of 1:20 (sperm: extender, v/v), then stored at 4 °C. The motility and viability were monitored every 24 h interval for 6 days. The results showed that the natural extender significantly affected the motility, viability, and sperm fertility of walking catfish (P<0.05). Young coconut water produced higher motility and viability on all observation days. The higher sperm motility, viability, and egg fertility on the last day of the experiment were also found in sperm diluted in young coconut water, with values of 45.83%, 43.83%, and 41.50%, respectively. These values were significantly different from other types of extenders. Therefore, it is concluded that young coconut water is the most suitable natural extender for sperm preservation of local catfish at 4 °C.

Myo-inositol improves the viability of boar sperm during liquid storage.

Liquid preservation of boar semen is a highly preferred method for semen preservation in pig production. However, oxidative stress is the main challenge during the liquid preservation of boar semen in a time dependent manner. Therefore, supplementation of sperm with antioxidants during storage to protect them from oxidative stress has been the focus of recent research. Myo-inositol (Myo-Ins), the most active form of inositol, which belongs to the vitamin (Vit.) (B1 group has been shown to improve semen quality) (1). This study aimed to investigate whether Myo-Ins supplementation protects boar sperm in liquid preservation against oxidative stress and determine the appropriate concentration of Myo-Ins to be used in this regard. Boar sperm was diluted with a semen extender with different concentrations of Myo-Ins (2, 4, 6, and 8 mg/mL) depending on the previous studies (1, 24). Sperm motility and viability, plasma membrane and acrosome integrity, mitochondrial membrane potential (MMP), semen time survival, and gene expression were measured and analyzed on days 0, 1, 3, 5, and 7 for the different samples. Different concentrations of Myo-Ins exerted different protective effects on the boar sperm quality. The addition of 2 mg/mL Myo-Ins resulted in higher sperm motility and viability, plasma membrane and acrosome integrity, MMP, and effective survival time. Investigation of mRNA expression patterns via qRT-PCR suggested that the 2 mg/mL Myo-Ins sample had increased expression of antioxidative genes. The addition of Myo-Ins to semen extender improved the boar semen quality by decreasing the effects of oxidative stress during liquid preservation at 17°C. Additionally, 2 mg/mL is the optimum inclusion concentration of Myo-Ins for semen preservation.

Application of the microflow technique for the selection of stallion semen

Routine semen preparation techniques are not always satisfactory for all stallions. Some stallions, despite poor semen parameters, are valuable for breeders who want to produce equally worthwhile offspring. In recent years, advanced sperm selection methods such as the microflow technique have been developed and successfully applied to humans. The aim of this study was to adapt the microflow technique for selection of stallion semen, focusing on their predicted benefits for sperm quality. The experiment was performed with semen of 6 stallions, which after combining in equal volumes created pooled research sample. Semen was left for 24 h in order to reduce the viability and motility of spermatozoa and divided into 3 parts, the control and the two research groups separated using ZyMōt Devices with two different incubation times (30 and 45 minutes). For each group, the basic sperm characteristics were assessed by SCA/CASA system, and fragmentation was evaluated using the TUNEL technique. The Z-score test was used for statistical analysis. In the sample incubated for 30 min in ZyMōt Devices, sperm motility increased from 28% to 52.15% (p=0.001), including sperm with progressive motility which increased from 8.47% to 41.91% (p<0.00001). After extending the incubation time for 45 minutes, these parameters were even higher: motility 62.43% (p<0.00001), progressivemovement 55.33% (p<0.00001). It should be noted, however, that in the case of both research samples, the concentration of sperm decreased from 804.68 M/ml in the control to 32.35 M/ml (p<0.00001) after 30 min incubation and 40.37 M/ml after 45 min incubation, respectively (p<0.0001). Sperm selection did not affect vitality. The TUNEL technique showed a very low percentage of spermatozoa with apoptosis in the semen fraction after selection at both incubation times. In summary, the microflow technique is an effective method for selection of stallion sperm with high motility, progressive movement and low DNA sperm fragmentation, with unchanged vitality parameters. Comparing the two incubation times, it was found that extending the incubation time to 45 minutes improved sperm concentration and spermatozoa with progressive motility. Nevertheless, prolonged incubation caused an increase in apoptotic spermatozoa that penetrate through a membrane with microchannels. It should be considered that sperm selection by microflow technique drastically reduces sperm concentration, which in practice translates into a significant reduction in the number of insemination doses which will increase costs significantly. However, this method can be successfully used for sperm selection for the ICSI procedure.

Buffalo anti-PDC-109 antibodies improve the semen quality profiles and in-vitro zona binding index and minimize the cryoinjury of sperm in cryopreserved buffalo semen

Objective: To optimize the concentration of PDC-109 protein in semen for higher cryopreservability and fertility by sequestration or neutralization of PDC-109 by its antibodies (anti-PDC-109 antibodies) in bubaline species. Methods: PDC-109 protein was purified by applying two-step chromatography procedures. Purified protein was injected in rabbits to raise antibodies. These raised anti-PDC-109 antibodies were used in neutralization or sequestration of PDC-109 in in-vitro model. Ejaculates were collected from buffaloes and splited for four groups. Group 1 received egg yolk Tris glycerol extender, without anti-PDC-109 antibodies, while group 2 to 4 received anti-PDC-109 antibodies 266 μg/mL, 80 μg/mL, and 26 μg/mL in Tris-fructosecitrate buffer, respectively. Semen quality parameters viz., forward progressive motility, viability, total morphological abnormality, acrosomal integrity, plasma membrane integrity, cryoinjury and in-vitro zona binding index were evaluated. Results: Semen quality parameters of neat semen were within the normal range of bubaline species. Sperm motility, livability, acrosomal integrity, plasma membrane integrity, and cholesterol content of sperm were decreased and total sperm abnormality was increased significantly in post-thaw semen compared to those in pre-freeze and fresh semen (P&lt;0.05). Semen in group 2 had higher sperm motility, livability, acrosomal integrity, plasma membrane integrity, and cholesterol content of sperm and lower total sperm abnormality significantly compared to those in group 1, 3 and 4 at pre-freeze and post-thaw stages (P&lt;0.05). Conclusions: Sequestration or neutralization of PDC-109 by its antibodies significantly improves pre-freeze, and post-thaw semen quality parameters and in-vitro zona binding index with simultaneously reducing cryoinjury or cryodamage in the sperm of bubaline species.