Sperm Selection Research Articles

Broiler breeder putative lipid biomarkers associated with sperm mobility

IntroductionBiomarkers indicative of sperm mobility in broiler breeders would provide the ability to screen for fertility potential, with a positive correlation established between sperm mobility and fertilization potential. This study characterized the lipidome of seminal plasma (SP), sperm cell (SC), and whole semen (WS) isolated from broiler breeder roosters with different sperm mobility phenotypes across key timepoints of the semen production cycle.MethodsWS samples were collected from five high mobility roosters and five low mobility roosters during early, mid, and late semen production, with SP separated from SC by centrifugation. Using multiple reaction monitoring (MRM) profiling, a total of 3241 lipid species were identified in rooster semen across ten lipid classes. Metaboanalyst 6.0 was used to analyze the relative ion intensity for each lipid species due to sperm mobility phenotype through a t-test and due to timepoint through a one-way ANOVA, with lipid ontology enrichment analysis performed using LION. Metaboanalyst 6.0 was also used to perform biomarker analysis for the sperm mobility phenotype in WS samples.ResultsLipid class total abundance differed with sample type, sperm mobility phenotype, and timepoint. A total of 31, 99, and 112 lipid species were found to be different between low and high mobility males across timepoints in the SP, SC, and WS samples, respectively. Lipid ontology enrichment analysis revealed stark contrasts in lipid-based functions key to sperm survival, storage, and productivity between low and high sperm mobility phenotypes. Through biomarker analysis, 8 lipid species were identified as excellent sperm mobility biomarkers that could be detected in early and mid-semen production.DiscussionTimepoint based changes in lipid species were unique to each sperm mobility phenotype, with low sperm mobility roosters exhibiting a larger number of lipid species changes over the semen production cycle in the SP and SC when compared to high sperm mobility roosters. This is the first study to characterize poultry semen lipidome using MRM profiling. The lipid species identified between low and high sperm mobility roosters could be utilized in the poultry industry as potential biomarkers of fertility potential, with the ability to screen for the economical trait of fertility potential early in semen production.

IVF with frozen-thawed sperm after prolonged capacitation yields comparable results to ICSI in horses: A morphokinetics study

Intracytoplasmic sperm injection (ICSI) is the current clinical practice for the in vitro production of equine embryos. The use of conventional fertilization methods such as in vitro fertilization (IVF), has historically been associated with poor success in horses. However, recent improvements have led to better outcomes with IVF, though only when using fresh semen, which limits its use in clinical practice. IVF remains in its infancy in equine reproduction, and several unknowns remain about the technique. One significant gap in knowledge concerns the morphokinetics of IVF embryos and how they differ from their ICSI counterparts. To address this, we performed IVF using frozen-thawed sperm from five different stallions following sperm selection and a prolonged capacitation period of 10 h, on a total of 109 oocytes. We then analyzed the cleavage rate (cleaved/initial oocytes), blastocyst rate (blastocyst/initial zygotes), and blastocyst development (blastocyst/cleaved zygotes) of the IVF cycles, and compared them with those of the clinical ICSI cycles during the same period. We also evaluated time-lapse images of the developed embryos to assess developmental time points such as time to morula compaction and blastocyst expansion, as well as morula and blastocyst sizes. Overall, developmental rates were not different between IVF and ICSI cycles (blastocyst rate 41.1 % IVF and 41.8 % ICSI, p > 0.05). However, development proceeded faster in IVF cycles (blastocyst expansion IVF 155.5 ± 18.5 h; ICSI 167.2 ± 19.6 h; p < 0.05) and IVF embryos were also larger (blastocyst area IVF 22608 ± 2857 μm2; ICSI 20806 ± 1505 μm2; p < 0.05). The faster development and larger size might suggest a more advanced developmental stage. The implications of these findings need to be further evaluated to assess their association with pregnancy potential. The successful developmental rates achieved in IVF cycles demonstrate the potential of this technique for clinical application, although the amount of frozen-thawed semen required is significantly higher in IVF than in ICSI, which is an important consideration for mare and stallion owners. Nonetheless, the use of frozen-thawed semen in equine IVF, coupled with comparable blastocyst rate, presents promising potential for broader clinical adoption of the IVF technique.

Anethole improves mitochondrial activity and quality parameters in fresh and frozen-thawed ovine semen

Anethole, an antioxidant found in plants, appears to improve the survival of spermatozoa during semen cryopreservation. This study assessed the effects of commercial trans-anethole in ram semen cryopreservation. Thirty ejaculates from six rams were diluted in media containing anethole at the following concentrations: CONT (0 μM), AN10 (10 μM), AN50 (50 μM), and AN100 (100 μM). Semen was slow-frozen, preserved in liquid nitrogen, and thawed. Anethole at 10 μM or 50 μM did not compromise any studied sperm quality parameter but increased pre-freezing functionality of membrane and mitochondrial activity. At 10 μM, anethole reduced post-thawing spermatozoa lipoperoxidation. At 50 μM, anethole sustained higher mitochondrial activity after thawing, reduced minor defects in sperm, and increased the number of sperm binding to perivitelline membrane, while keeping lipoperoxidation levels as in control. Anethole at 100 μM promoted higher pre- and post-freezing mitochondrial activity and higher number of sperm binding to perivitelline membrane, in comparison to control. Additionally, some post-thawing kinematic parameters were enhanced by anethole at 100 μM. Of note, mitochondrial activity and lipoperoxidation were higher with anethole at 100 μM in comparison to 50 μM, not differing from control. At the hypoosmotic test, the highest concentration (100 μM) tested reduced sperm osmotic resistance. The results of this study indicate that using anethole in cryopreservation media promoted mostly positive effects on the fresh and post-thawed ram semen, and the advantages vary according to its concentration.

Transcriptome and Metabolome Analyses Reveal High-Altitude Adaptation Mechanism of Epididymis Sperm Maturation in Tibetan Sheep.

In this study, the epididymal histology, caepididymal sperm physiological parameters, serum reproductive hormones, and antioxidant enzyme SOD levels of Tibetan sheep at a 2500 m and 3500 m altitude were compared by using a combination of transcriptome and metabolomics methods. This was conducted to investigate the effects of a high-altitude environment on spermatogenesis and the maturation of Tibetan sheep. The results showed that compared to the low-altitude group, the high-altitude group had a smaller epididymal lumen, thicker epididymal wall, significantly decreased sperm survival rate, and significantly increased sperm deformation rate, but no difference in sperm motility and sperm respiratory intensity. With increasing altitude, Tibetan sheep showed a decreasing trend in serum reproductive hormones (FSH and T), while the antioxidant enzyme SOD activity was significantly reduced. Transcriptomic analysis revealed 139 differentially expressed genes in the Tibetan sheep epididymis under high-altitude conditions. The SYCP2 gene is involved in multiple biological processes related to reproduction and plays an important role in the regulation of epididymal function and sperm quality in Tibetan sheep. Genes like ADCYAP1R1, CABP2, CALN1, and ATP6V1B1 can help maintain sperm viability and maturation by regulating the cAMP signaling pathway, calcium ion homeostasis, and cellular signaling. Metabolomic analysis found that the high-altitude group had increased adenosine content and decreased prostaglandin I2 content in the epididymis. These metabolites are involved in spermatogenesis, motility, fertilization, and early embryonic development. The integrated omics analysis suggests that Tibetan sheep adapt to the high-altitude hypoxic environment by regulating cAMP signaling pathway genes like ADCY and PRKACA, as well as metabolites like adenosine and prostaglandin I2, to maintain epididymal function and sperm motility. These genes and metabolites play an important role in maintaining normal epididymal function and sperm motility at high altitudes.

Control of sperm penetration using stereumamide A derived from Trichaptum fuscoviolaceum in the in vitro fertilization of pig oocytes.

Fungal metabolites are known to have potent and diverse properties such as antiviral, antidiabetic, antitumour, antioxidant, free radical scavenging, and antibacterial effects which can be utilized to treat diseases. In this study, we investigated the functional activity of stereumamide A (StA) derived from a culture broth of Trichaptum fuscoviolaceum during the in vitro fertilization (IVF) of pig oocytes, to determine its effects on sperm penetration. Oocytes matured in vitro were fertilized in the absence or presence of varying concentrations of StA (0-50 μg/ml StA). When StA was directly added into the IVF medium, significantly lower fertilization rates were seen with the 20 or 50 μg/ml StA (2.0-17.5%) treatments compared with those of 10 μg/ml StA or the controls (60.9-62.3%), whereas StA had no influence on the survival of oocytes and spermatozoa throughout the IVF process. For evaluating the control of sperm entry, mature oocytes were pre-incubated in a medium containing 20 μg/ml StA for 1 h, and then IVF was subsequently performed. The incidence of polyspermy was significantly reduced when oocytes were pre-incubated with StA (15.0% vs. 50.4-57.5% in controls). In conclusion, sperm penetration was inhibited in the medium in the presence of StA during IVF, while StA did not affect sperm motility and fertility competence. Fertilization was controlled when mature oocytes were incubated with StA prior to IVF, suggesting the possible use of the fungal metabolite in assisted reproductive technology for humans and animals.

Benchmarks defining high-quality sperm in the common marmoset (Callithrix jacchus).

Common marmosets (Callithrix jacchus) are increasingly recognized as valuable nonhuman primates (NHPs) for biomedical research due to their small size and short reproductive cycle and lifespan relative to other NHP species. Maximizing the utility of captive research marmosets, including genetically manipulated animals, will require the use of assisted reproductive techniques (ART) including manipulation, storage, and sharing of marmoset sperm. Here, we identify characteristics of high-quality semen samples and validate a simple method for selecting high-quality sperm. Computer-assisted sperm analysis (CASA) was used to evaluate sperm quality in semen samples collected from 44 marmosets and assessed the use of the swim-up method for the selection of high-quality sperm was also tested in half the samples as a potential means to optimize in vitro fertilization or intrauterine insemination. For each reference parameter, samples at or below the 5th percentile were categorized as abnormal sperm, while those above the 5th percentile were considered to be normal. Among normal samples, those at or above the 50th percentile were categorized as high-quality. High-quality semen samples exhibited the following characteristics: semen volume ≥ 30µL; sperm count ≥ 107/ejaculate; total motility ≥ 35%; and normal morphology ≥ 5%. Sperm isolated by swim-up exhibited superior sperm progressive motility (19.7%±4.5vs. 5.6%±2.1; P=0.01) and normal morphology (13.1±1.59vs. 7.65±1.1; P<0.001) compared with unselected sperm. This study defines robust, statistically supported reference values for evaluating marmoset semen samples to assist with the identification of optimal sperm donors and the selection of high-quality sperm samples for assisted reproduction. Ultimately, these reference values combined with a validated selection method will contribute to consistent standards for the international sharing of genetically diverse and/or gene-edited marmoset sperm for research and reproduction.

Sperm histone modifications may predict success in human assisted reproduction: a pilot study.

Currently, assisted reproduction clinics employ various sperm selection techniques to identify the best sperm for fertilization. However, these techniques may not assess crucial sperm traits that can substantially impact embryonic quality. To address this, we propose analyzing diverse histone modifications as potential markers of sperm functionality and success in assisted reproduction techniques. Cross-sectional pilot study including infertile male patients attending an infertility clinic in CABA, Argentina between April and August 2019 was performed. We used immunofluorescence techniques to evaluate post-translational modifications of histones in sperm and established correlations with in vitro fertilization outcome and embryo quality. Our findings indicate a negative correlation between H3K4me3 and H3K4me2 marks and fertilization rate and showed a positive correlation of this parameter with H3K9me mark. In addition, there was a positive correlation between H3K27me3 and good embryo quality. This pilot study proposes a non-invasive strategy to predict embryo quality by analyzing spermatozoa prior to fertilization. The assessment of histone post-translational modifications in sperm samples could provide useful information for the recognition of epigenetic marks that could predict the health of the embryo of an assisted fertilization treatment.

Therapeutic Effects of Proanthocyanidins on Diabetic Erectile Dysfunction in Rats.

The rising occurrence of erectile dysfunction related to diabetes mellitus (DMED) has led to the creation of new medications. Proanthocyanidins (PROs) is a potential agent for DMED. In this study, the DMED rat model was established using streptozotocin (STZ) and erectile function was assessed using apomorphine (APO) in rats. Following this, the rats were subjected to oral treatment with PRO. Then, we evaluated the influence of PROs on DMED rats. The findings suggest that PROs significantly enhance erectile function in DMED rats. PROs modulated glucose and lipid metabolism in DMED rats by decreasing blood glucose and lipid levels while increasing liver glycogen and serum insulin levels. Furthermore, PROs enhanced vascular endothelial function in DMED rats by augmenting nitric oxide (NO) levels and reducing the levels of endothelin-1 (ET-1) and lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1). Additionally, PROs have been shown to elevate testosterone (T) levels, mitigate pathological testicular damage, and enhance sperm concentration and survival rates. Finally, the core targets were screened using network pharmacology, followed by validation through molecular docking, enzyme-linked immunoassay (ELISA), and real-time PCR methodologies. Our findings imply that PROs may treat DMED by elevating AKT1 levels while concurrently diminishing CASP3 levels, thereby effectively regulating the PI3K-Akt signaling pathway. Overall, these results support using PROs as a potential candidate for the treatment of DMED.

Microfluidics-A novel technique for high-quality sperm selection for greater ART outcomes.

Microfluidics represent a quality sperm selection technique. Human couples fail to conceive and this is so in a significant population of animals worldwide. Defects in male counterpart lead to failure of conception so are outcomes of assisted reproduction affected by quality of sperm. Microfluidics, deals with minute volumes (μL) of liquids run in small-scale microchannel networks in the form of laminar flow streamlines. Microfluidic sperm selection designs have been developed in chip formats, mimicking invivo situations. Here sperms are selected and analyzed based on motility and sperm behavioral properties. Compared to conventional sperm selection methods, this selection method enables to produce high-quality motile sperm cells possessing non-damaged or least damaged DNA, achieve greater success of insemination in bovines, and achieve enhanced pregnancy rates and live births in assisted reproduction-in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). Besides, the concentration of sperm available to oocyte can be controlled by regulating the flow rate in microfluidic chips. The challenges in this technology are commercialization of chips, development of fully functional species-specific microfluidic tools, limited number of studies available in literature, and need of thorough understanding in reproductive physiology of domestic animals. In conclusion, incorporation of microfluidic system in assisted reproduction for sperm selection may promise a great success in IVF and ICSI outcomes. Future prospectives are to make this technology more superior and need to modify chip designs which is cost effective and species specific and ready for commercialization. Comprehensive studies in animal species are needed to be carried out for wider application of microfluidic sperm selection in invitro procedures.

Comparison of different processed products of Allium tuberosum Rottler for the treatment of mice asthenozoospermia.

Allium tuberosum Rottler improves sexual function and is used in the treatment of impotence and spermatorrhea. However, its chemical composition and mechanism of action remain unclear. This study investigates the chemical composition and mechanism of action of Allium tuberosum Rottler co-processed with salt and wine (GZP) in modulating testicular mitochondrial autophagy for the treatment of asthenozoospermia in mice. Adenine gavage + cyclophosphamide intraperitoneal injection was used to establish the model of asthenozoospermia, and six Allium tuberosum Rottler processed products were compared in the pharmacological efficacy for the treatment of asthenozoospermia in mice. The liquid chromatograph mass spectrometer (LC-MS) assay was performed to analyse the compositional changes in the GZP. The mechanism of GZP in the treatment of asthenozoospermia in mice was further investigated. The mitophagy was detected by transmission electron microscope (TEM) and immunofluorescence, respectively. Reactive oxygen species (ROS) were detected by probe. Protein expression was determined by Western blotting. GZP exhibited optimal therapeutic effects on asthenozoospermia in mice. It showed the best therapeutic effect in improving the total number of spermatozoa, sperm survival rate, improving sperm viability and reducing sperm deformity rate, alleviating the abnormal pathological morphology of mice testis, and increasing the serum testosterone (T), follicle-stimulating hormone (FSH) and prolactin (PRL) levels in mice. The LC-MS detection found that Allicin showed the most significant increase in GZP. Besides, GZP reduced ROS level and inhibited mitophagy in mice testicular tissues. Meanwhile, it restrained the expression of PINK1, Parkin, Light chain 3II (LC3-II)/Light chain 3I (LC3-I) and Caspase-3 proteins. GZP improves asthenozoospermia via inhibiting excessive mitophagy and protects the integrity of mitochondria by blocking the PINK1/Parkin signaling pathway. During which, the Allicin may play an important role.

Comparative study of Tris Turmeric, Tris Turmeric Dimethyl Sulfoxide and Tris Turmeric Ethylene glycol extenders on the cryosurvivability, sperm resistance, in- vivo fertility and antioxidant status in buffalo bull semen

The freeze-thaw process leads to structural and functional damage due to excessive accumulation of reactive oxygen species (ROS). The addition of exogenous antioxidants to sperm diluents is of great importance to overcome oxidative damage during freezing. The purpose of this study was to explore the effects of three diluents Tris Turmeric, Tris Turmeric Dimethyl Sulfoxide and Tris Turmeric Ethylene glycol on the cold survival ability of buffalo sperm. Semen was collected from five local adult male buffalo breeds. A base diluent of Tris-citric acid-fructose (TCF) was prepared, adding 20% whole egg yolk (TCFY). The Tris extender without turmeric, without DMSO, and without EG was kept as a control. Other extenders are Tris containing turmeric TT (100 ml/5 ml Tris), Tris containing turmeric dimethyl sulfoxide TTD (100 ml/5 ml Tris + 1.5% DMSO) and Tris containing turmeric and ethylene glycol TTE EG (100 ml/5 ml Tris + 1.5% EG). Semen samples were added and a pure sperm concentration of 60 × 106/ml was achieved. Frozen buffalo sperm after thawing showed significant improvements in all research parameters of the three breeding samples compared to the control. Tris Turmeric Ethylene was the type that best improved sperm survival under frozen conditions, followed by Tris Turmeric and Tris Turmeric Dimethyl Sulfoxide compared to the control. A significant decrease in sperm motility after thawing was evident as usage time increased in all expanders. There was a significant increase in total antioxidant content (TAC) and insignificant change in malondialdehyde (MDA) of the diluent used compared to the control. Conception rate (CR) was higher in Tris Turmeric Ethylene glycol (65.2%), followed by Tris Turmeric (60.3%) and Tris Turmeric Dimethyl Sulfoxide (55.9%) compared to the control (36, 7%). It can be concluded that Tris Turmeric Ethylene Glycol is considered the best agent for improving cold survival and sperm fertility, followed by Tris Turmeric and Tris Turmeric Dimethyl Sulfoxide.

Ascorbic acid 2-glucoside improves survival, quality, and fertility of frozen-thawed C57Bl/6J and C57Bl/6N mouse spermatozoa.

Ascorbic acid 2-glucoside (AA2G) is a stabilized form of ascorbic acid and a potent antioxidant. Ascorbic acid is present in the testes and epididymis and helps maintain the physiological integrity of reproductive organs. Its properties have been utilized to protect spermatozoa of different species from oxidative stress. Spermatozoa of C57Bl/6J and C57Bl/6N strains were frozen and analyzed, after thawing, by supplementing the capacitation medium with AA2G, both in the presence and absence of methyl-β-cyclodextrin (MBCD). The effect of treatment was evaluated by SCA System (Microptic) analyzing the velocity, vitality, morphology, and the DNA fragmentation. We also examined sperm capacitation (CTC), acrosome reaction (Coomassie Brillant Blue), and fertility (in vitro fertilization) of treated spermatozoa. AA2G improved sperm quality and fertility particularly in association with MBCD. We observed a significant increase of sperm motility, velocity, and vitality associated with an enhanced capacitation and acrosome reaction. These improvements resulted in a marked increase in in vitro fertilization success. Embryos obtained were cultured and reached normally the blastocyst stage. This study aimed to determine if AA2G could safeguard mouse spermatozoa during cryopreservation. We found a protective effect of AA2G that increased sperm survivability resulting in higher fertilization rate. This newly improved protocol shows potential for reanimating cryopreserved GEMMs stored in mouse biobanks and international repositories, such as the European Mouse Mutant Archive (EMMA). This can serve as a pivotal tool in fulfilling the 3Rs mission (replacement, reduction, and refinement), promoting ethical and humane research practices.

Predicting Boar Sperm Survival during Liquid Storage Using Vibrational Spectroscopic Techniques.

Artificial insemination (AI) plays a critical role in livestock reproduction, with semen quality being essential. In swine, AI primarily uses cool-stored semen adhering to industry standards assessed through routine analysis, yet fertility inconsistencies highlight the need for enhanced semen evaluation. Over 10-day storage at 17 °C, boar semen samples were analyzed for motility, morphology, sperm membrane integrity, apoptosis, and oxidative stress indicators. Additionally, machine learning tools were employed to explore the potential of Raman and near-infrared (NIR) spectroscopy in enhancing semen sample evaluation. Sperm motility and morphology gradually decreased during storage, with distinct groups categorized as "Good" or "Poor" survival semen according to motility on Day 7 of storage. Initially similar on Day 0 of semen collection, "Poor" samples revealed significantly lower total motility (21.69 ± 4.64% vs. 80.19 ± 1.42%), progressive motility (4.74 ± 1.71% vs. 39.73 ± 2.57%), and normal morphology (66.43 ± 2.60% vs. 87.91 ± 1.92%) than their "Good" counterparts by Day 7, using a computer-assisted sperm analyzer. Furthermore, "Poor" samples had higher levels of apoptotic cells, membrane damage, and intracellular reactive oxygen species on Day 0. Conversely, "Good" samples maintained higher total antioxidant capacity. Raman spectroscopy outperformed NIR, providing distinctive spectral profiles aligned with semen biochemical changes and enabling the prediction of semen survival during storage. Overall, the spectral profiles coupled with machine learning tools might assist in enhancing semen evaluation and prognosis.

Sperm Storage and Use Following Multiple Insemination in Aedes albopictus: Encouraging Insights for the Sterile Insect Technique.

The key to success in the application of the sterile insect technique (SIT) relies on the ability of released, sterile males to outcompete their fertile wild male counterparts to mate with wild females. However, many insect species exhibit multiple-mating behavior, which can be a way for females to select paternity for their progeny. This study aims to recognize the consequences of potential double-matings during an SIT program and to detect any evidence of sperm selection favoring sperm from fertile mates. This report provides a descriptive analysis of the storage and use of sperm by female Aedes albopictus. Stable isotopes were used to mark the sperm of fertile and sterile males. Mated females were allowed to oviposit before dissecting the spermathecae to link the presence of each type of sperm to the sterility of the eggs laid. It was found that sperm in females inseminated by both males was distributed in the three spermathecae with no obvious pattern, mostly mixed but also separately, and no evidence of any mechanism for sperm selection, sperm precedence, or sperm competition in Ae. albopictus females could be found. The fact that only a few double-mated females were double-inseminated and could also produce semi-sterile eggs, together with the finding that the sperm of sterile males appeared to be no less viable than that of fertile males, is an encouraging outcome for SIT approaches.

Application of Artificial Intelligence in Sperm Quality Analysis and Sperm Screening

Infertility is a global health issue, and more and more people are hoping to have babies by means of assisted reproductive technology. However, there are still many challenges in fertilization and pregnancy outcomes. Sperm quality is a key factor affecting the success rate of assisted reproduction. Therefore, sperm quality screening is crucial for achieving breakthroughs in assisted reproduction technology. At present, with its capabilities in the field of image recognition, artificial intelligence (AI) is providing new ideas and methods for sperm screening. Various attempts have been made with AI-based models to evaluate indicators such as sperm morphology, DNA quality, and motility level, and some results have been achieved. Herein, we reviewed the application of AI in sperm quality analysis and selection, providing support for the future development of AI and the improvement in the fertilization rate and outcomes of assisted reproductive technology.

Live Motile Sperm Sorting Device Improves Embryo Aneuploidy: A Retrospective Cohort Study

Background: Conventional sperm selection methods, involving centrifugation, exert a detrimental effect on sperm DNA integrity due to mechanical stress. The recent noninvasive sperm selection device, CA0, based on live sperm sorting technology, facilitates the retrieval of highly motile sperm and minimizes sperm DNA fragmentation (SDF). This study was to investigate the impact of various sperm separation methods, with and without centrifugation, on embryo ploidy status. Methods: The retrospective study comprised 82 intracytoplasmic sperm injection (ICSI) cycles involving preimplantation genetic testing for aneuploidy (PGT-A) cases, with a focus on recruiting egg donation cycles to thoroughly investigate the impact of male factors. Two populations are classified based on semen quality: normozoospermic ([Formula: see text] = 33) and non-normozoospermic ([Formula: see text] = 49). Subjects were allocated to either swim-up (SU) or CA0. Preimplantation genetic testing results were recorded. Results: When comparing male characteristics between subgroups, no significant differences were observed except for a lower normal morphology rate in the CA0 group compared to SU (SU: 3 [3–4] vs. CA0: 2 [2–2.8], [Formula: see text] &lt; 0.0001) in the non-normozoospermic cohort. There were no differences in female factors such as age and mature oocyte count (MII) number between subgroups, indicating that this model is ideal for assessing the impact of male factors on clinical outcomes. In the normozoospermic cohort, euploidy rates were similar between SU and CA0 (SU: 71.9% vs. CA0: 64.2%). However, in the non-normozoospermic cohort, CA0 showed a significantly higher euploidy rate compared to SU (SU: 53.6% vs. CA0: 74.2%) and a lower aneuploidy rate (SU: 37.1% vs. CA0: 25.8%). Additionally, CA0 minimized the incidence of mosaic embryos, whereas a mosaicism rate of 9.3% was observed with SU. This trend highlights CA0’s distinct advantage in optimizing outcomes for non-normozoospermic cases. Conclusions: CA0 is a reliable intervention to optimize paternal genetic quality before assisted insemination, thereafter effectively reducing the incidence of embryo aneuploidy associated with male factors.

Effect of Two Different Sperm Selection Methods on Boar Sperm Parameters and In Vitro Fertilisation Outcomes.

Porcine in vitro embryo production (IVP) protocols have conventionally used density gradient selection (DGS) by centrifugation to prepare sperm samples and achieve successful fertilisation. However, the possible toxicity of the solutions used and the potential damage caused by the centrifugation step may have a negative effect on the quality of the sample. Microfluidic chip-based sperm (MCS) sorting has been proposed as an alternative technique for the selection of high-quality sperm with the purpose of improving reproductive outcomes in IVF. This device does not require centrifugation or any toxic solution to prepare the sample for fertilisation. The sample is not subjected to unnecessary stress, and the process is less operator-dependent. In this study, we compared the sperm parameters of unselected extender-diluted boar semen samples with selected samples using DGS and MCS methods. The results show an expected reduction in sperm concentration after both methods. All the groups were significantly different from one another, with MCS being the group with the lowest concentration. Though the three groups had a similar overall motility, significant differences were found in progressive motility when comparing the unselected group (control, 19.5 ± 1.4%) with DGS and MCS. Progressive motility in DGS was also significantly higher than in MCS (65.2 ± 4.9% and 45.7% ± 5.3, respectively). However, MCS selection resulted in enriched sperm samples with a significantly lower proportion of morphologically abnormal sperm compared to DGS. After fertilisation, no statistical differences were found between the two methods for embryological parameters such as cleavage rates, blastulation rates, and embryo quality. The number of cells in blastocysts derived from MCS was significantly greater than those derived from DGS sperm. Thus, we demonstrate that MCS is at least as good as the standard DGS for most measures. As a more gentle and reproducible approach for sperm selection, however, it could improve consistency and improve IVP outcomes as mediated by a greater proportion of morphologically normal sperm and manifested by a higher cell count in blastocysts.

Sperm Selection Techniques for ART: Light at the End of the Tunnel?

Sperm Selection Techniques for ART: Light at the End of the Tunnel?

Dietary alpha-linolenic acid supplementation enhances semen quality, antioxidant capacity, and sperm survival in aging breeder roosters

Aging in breeder roosters is often accompanied by a decline in semen quality, negatively impacting reproductive performance. This study aimed to investigate the effect of dietary alpha-linolenic acid (ALA), an essential omega-3 polyunsaturated fatty acid, on semen quality, antioxidant capacity, and sperm survival in aging breeder roosters. Roosters were divided into 4 groups and fed diets supplemented with 0%, 0.5%, 1%, and 2% ALA for 6 wk. Results indicated significant improvements in semen volume, sperm viability, and sperm density in ALA-supplemented groups compared to the control (P < 0.05). The 1% ALA group exhibited the most notable enhancements in sperm viability and density. Additionally, ALA supplementation increased the activities of antioxidant enzymes such as superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and reduced malondialdehyde (MDA) levels, indicating enhanced antioxidant capacity (P < 0.05). Furthermore, ALA improved mitochondrial membrane potential (MMP) and reduced early and late sperm apoptosis, with the 2% ALA group showing the highest MMP and the lowest ROS-positive rate (P < 0.05). These findings suggest that dietary ALA supplementation enhances semen quality and antioxidant defenses, and mitigates oxidative stress, thus supporting the reproductive health of aging breeder roosters. This study underscores the potential of ALA as a dietary strategy to improve reproductive efficiency in poultry production.

Effect of seminal plasma cholesterol and triacylglycerides concentrations and sperm morphology on semen freezability in domestic cats (Felis silvestris catus)

There is scarce information about the effect of sperm morphology and seminal plasma composition on cat semen freezability. Thus, this study aims to assess the effect of cat sperm morphology and seminal plasma cholesterol (CHOL) and triacylglyceride (TAG) concentrations on sperm post-thaw survival. Ejaculates (n = 49) were evaluated, and seminal plasma was separated and frozen until CHOL and TAG concentrations were measured. The sperm pellet was diluted in a tris-based egg yolk extender, frozen (n = 38), or processed for sperm ultrastructure study (n = 11). Abnormalities recorded were abnormal head shape and size, detached heads, knobbed or ruffled acrosomes, eccentric mid-piece insertion, proximal and distal cytoplasmic droplets, folded and coiled tails, and Dag defect. Ultramicroscopic evaluation detected several sperm abnormalities in fresh semen and some sperm damage in frozen semen. Seminal plasma lipids components were positively correlated with post-thaw motility and acrosome integrity. Higher freezability indices for motility and acrosome integrity were observed in frozen-thawed semen with high seminal plasma CHOL and TAG concentrations. No freezability differences were observed between teratozoospermic and normozoospermic ejaculates. Our results showed that even when seminal plasma was removed before cryopreservation, sperm survival after thawing was significantly higher in samples with high seminal plasma CHOL and TAG concentrations, indicating a rapid adherence to these compounds to the sperm plasma membrane, protecting sperm cells from temperature changes. Nevertheless, there were no differences in sperm freezability by sperm morphology.