Relationship of motility and acrosome reaction on sexing sperm in Ongole Crossbred bull

Artificial insemination using sexing semen is expected to produce calves with the desired sex. One sexing sperm method is the percoll density gradient centrifugation method. This study aimed to determine the changes and the relationship between motility and acrosome reaction after sexing process using percoll density gradient centrifugation. The material used was semen of ±5 years old Ongole crossbred bull with a bodyweight of ±700 kg as many as three bulls with mass motility 2+ and individual motility 70%. The method used was to compare fresh semen with sexed semen after the cooling process. Parameters measured were motility characters using CASA analysis, which included motility parameters, progressive motility, capacitation, and no acrosome reaction. Statistical analysis used paired T-test to distinguish among fresh semen, after sexing and cooling process. In comparison, regression and correlation were used to analyze the relationship of capacitation and hyperactivation sperm with no acrosomal reaction with motility and progressive motility. The results showed that motility and progressive motility decreased after the sexing and cooling process. Meanwhile, the acrosomal reaction, capacitation, and hyperactivity increased.

The Use of κ-Carrageenan in Egg Yolk Free Extender Improves the Efficiency of Canine Semen Cryopreservation

κ-Carrageenan is a plant polysaccharide derived from red seaweeds reported to possess potential medicinal and antioxidants activities. The present study aimed to identify the cryoprotective effects of κ-carrageenan on the quality of frozen-thawed canine semen. Twenty-eight ejaculates were collected and diluted in a Tris egg-yolk-free extender supplemented with various concentrations of κ-carrageenan (0.0%, 0.1%, 0.2%, 0.3%, and 0.5%). The addition of κ-carrageenan to the extender at a 0.2% concentration induced a significant increase in the total motility (TM) and the rapid progressive motility (RPM) of canine sperm. Among the experimental groups, the highest percentage of sperms with intact acrosomes was found in the 0.5% κ-carrageenan group (p < 0.05). Apoptosis levels were significantly lower in the 0.1% and 0.2% κ-carrageenan treatment. Moreover, sperm in the κ-carrageenan supplemented group showed a significantly higher expression of antiapoptotic (Bcl-2) and lower expression of NADPH oxidase (NOX5), spermine synthase (SMS), and spermine oxidase (SMOX) genes than those in the control group. In conclusion, the addition of κ-carrageenan to the freezing extender improved the overall efficiency of frozen-thawed dog spermatozoa.

Effect Concentration of Moringa (Moringa oleifera Lam) Leaf Extract in Citrate-Egg Yolk in Maintaining Motility and Viability of Spermatozoa of Kacang Goat

This study aimed to determine the effectiveness and the best concentration of Moringa leaf extract (MLE) in the citrate-egg yolk (C-EY) to maintain the motility and viability of spermatozoa kacang goat. Semen was collected from 3 goats aged two years; by using the artificial vagina method. The semen was evaluated macroscopically and microscopically. The semen that had >70% sperm motility and >250x106/ml sperm concentration was divided into 4 equal tubes, each diluted with100% C-EY (P1), 10% MLE+ 90%C-EY (P2), 20% MLE +80% C-EY (P3), and 30% MLE+70% C-EY (P4). The diluted samples were then stored in a refrigerator (3-5?C) and evaluated for motility and viability every 24 hours. The study was designed using a completely randomized design (CRD) consisting of four treatments and five replications. The results showed that the addition of MLE in C-EY significantly affected goat spermatozoa's progressive motility and viability. The data showed that the spermatozoa kept during four days in a diluent of P2 had higher (P<0.05) motility 44.67±4.80% and viability 74.24±4.46%than the other three diluents of P1(36.00±4.70%; 70.10±3.6%), P3(33.67±0.42%; 66.85±4.99%) and P4 (29.67±3.99%; 63.96±5.44%). This study concluded that adding 10% MLE was the best concentration as source energy in 90% C-EY diluents, which effectively maintained the motility and viability of kacang goat spermatozoa for four days of storage at a temperature of 3-5oC.

Sperm DNA 5-mC and RNA m 6A methylation are differently affected during periods of body weight losses and body weight gain of young and mature breeding bulls

Aiming to characterize the effects of nutritional status on epigenetic markers, such as DNA 5-mC methylation and RNA m 6A methylation, of bovine sperm, twelve Angus × Hereford crossbred breeding bulls were submitted to nutritional changes for a period of 180 days: no change in BW (Phase 1 = 12 d); BW loss (Phase 2 = 78 d); and BW gain (Phase 3 = 90 d) in a repeated measures design. Animals were fed Beardless wheat (Triticum aestivum) hay and mineral mix. Statistical analyses were performed using SAS 9.4 (SAS Inst., Cary, NC). Higher levels of RNA m 6A (P = 0.004) and DNA methylation (P = 0.007) of spermatic cells were observed at Phase 2 compared with Phase 1. In Phase 3, sperm RNA m 6A methylation levels continued to be higher (P = 0.004), whereas the DNA of sperm cells was similar (P = 0.426) compared with the Phase 1. Growing bulls had a tendency (P = 0.109) of higher RNA m 6A methylation levels than mature bulls. Phase 2 altered scrotal circumference (P &lt; 0.001), sperm volume (P = 0.007), sperm total motility (P = 0.004), sperm progressive motility (P = 0.004), total sperm count (P = 0.049), normal sperm (P &lt; 0.001), abnormal sperm (P &lt; 0.001), primary sperm defects (P = 0.039), and secondary sperm defects (P &lt; 0.001). In Phase 3, bulls had scrotal circumference, sperm volume, sperm motility, sperm progressive motility, total sperm count, normal and abnormal spermatozoa, and primary and secondary spermatozoa defects similar to Phase 1 (P &gt; 0.05). Serum concentrations of insulin-like growth factor-1 (IGF-1) and leptin decreased during Phase 2 (P = 0.010) while no differences (P &gt; 0.05) were detected between Phase 3 and 1; growing bulls tended (P = 0.102) to present higher leptin levels than mature bulls. Specific for mature bulls, DNA methylation was positively correlated with leptin concentration (0.569, P = 0.021). Whereas for young bulls, DNA methylation was positively correlated with abnormal spermatozoa (0.824, P = 0.006), primary spermatozoa defect (0.711, P = 0.032), secondary spermatozoa defect (0.661, P = 0.052), and negatively correlated with normal spermatozoa (−0.824, P = 0.006), total sperm count (−0.702, P = 0.035), and sperm concentration (−0.846, P = 0.004). There was no significant correlation (P &gt; 0.05) between RNA m 6A and hormones and semen traits. In conclusion, the nutritional status of breeding bulls alters epigenetic markers, such as DNA methylation and RNA m 6A methylation, in sperm, and the impact of change seems to be age-dependent. These markers may serve as biomarkers of sperm quality and fertility of bulls in the future. Detrimental effects on sperm production and seminal quality are observed at periods and places when and where environmental and nutritional limitations are a year-round reality and may carry hidden players that may influence a lifetime of underperformance.

Using artificial intelligence to predict the intrauterine insemination success rate among infertile couples

Background To evaluate the use of artificial intelligence (AI) in predicting the success rate of intrauterine insemination (IUI) treatment among infertile couples and also to determine the importance of each of the parameters affecting IUI success. This study was a retrospective cohort study in which information from 380 infertile couples undergoing IUI treatment (190 cases resulting in positive pregnancy test and 190 cases of failed IUI) including underlying factors, female factors, sperm parameters at the beginning of the treatment cycle, and fertility results were collected from 2013 to 2019 and evaluated to determine the effectiveness of AI in predicting IUI success. Results We used the most important factors influencing the success of IUI as a neural network input. With the help of a three-layer neural network, the accuracy of the AI to predict the success rate of IUI was 71.92% and the sensitivity and specificity were 76.19% and 66.67%, respectively. The effect of each of the predictive factors was obtained by calculating the ROC curve and determining the cut-off point. Conclusions The morphology, total motility, and progressive motility of the sperm were found to be the most important predictive factors for IUI success. In this study, we concluded that by predicting IUI success rate, artificial intelligence can help clinicians choose individualized treatment for infertile couples and to shorten the time to pregnancy.

Relevance of the Ejaculate Fraction and Dilution Method on Boar Sperm Quality during Processing and Conservation of Seminal Doses

During boar semen processing and distribution, maximizing the work protocols in the laboratories becomes essential for the conservation of seminal doses. One of the recent implementations in the boar studs to improve efficiency has been semi-automatic semen collection systems, which do not allow to discard fractions of the ejaculate. The objective of this work was to evaluate the dilution method and vibrations (simulating delivery transport) effect on sperm quality (motility, viability, morphology, thermo-resistance test) according to the fraction of ejaculate collected. Two different fractions of the ejaculate were obtained [rich fraction (RF); total fractions (TF)] from six boars, and two dilution methods applied [pouring the extender over the semen (control; ES); pouring the semen over the extender (reverse; SE)]. The seminal doses (2000 × 106 sperm/50 mL) were preserved for 5 days. The results showed that the fraction collected affects sperm quality (better total and progressive motility, and faster sperm in TF; p < 0.05) regardless of the dilution method applied. However, these differences diminished after submitting the semen to the thermo-resistance test, with only differences in sperm viability being observed (p < 0.05). When seminal doses were subjected to vibrations, the sperm viability was more affected in the TF than in the RF group (p < 0.05). In conclusion, using the TF ejaculate leads to comparable results to the RF in sperm quality during storage regardless of the dilution method applied. However, the vibrations of seminal doses are more affected in doses prepared with TF than with RF, although more factors should be included to approach the real conditions during transport.

Effect of Cold Storage on Livability and Progressive Motility of Cauda Epididymal Spermatozoa of Red Sokoto Buck Recovered Post Mortem

Post mortem spermatozoa recovery is an important tool for harvesting germplasm. The study aimed to determine the effect of cold storage time at 4°C for 48 and 72 hours on the quality of sperm of Red Sokoto buck caudal epididymaly spermatozoa in terms of motility and livability recovered postmortem. Fifteen testicles with attached epididymis were collected and processed at 0, 48, and 72 hours of storage. The samples were divided into 3 groups according to the time processing after collection ; 0 hour, 48 hours and 72 hours, with a total of 5 samples per group. Each storage group motility and livability was evaluated, mean and the standard error of their mean was recorded. Mean percent sperm motility declined significantly ( p< 0.05) as postmortem storage time increased ( motility: 79.00+/- 2.92%, 40.00+/- 4.18, 37.00+/- 3.39) .There was no significant (p> 0.05) decrease in livability from the 0 hour storage to the 48 hour and 72 hours storage ( 75.60+/- 4.40, 73.60+/- 3.84, 68.20+/- 2.51). The current study has shown that viable spermatozoa can be recovered after 48 hours of storage at 4˚C of the cauda epididymides Red Sokoto buck.

Sperm preparedness and adaptation to osmotic and pH stressors relate to functional competence of sperm in Bos taurus

The adaptive ability of sperm in the female reproductive tract micromilieu signifies the successful fertilization process. The study aimed to analyze the preparedness of sperm to the prevailing osmotic and pH stressors in the female reproductive tract. Fresh bovine sperm were incubated in 290 (isosmotic-control), 355 (hyperosmotic-uterus and oviduct), and 420 (hyperosmotic-control) mOsm/kg and each with pH of 6.8 (uterus) and 7.4 (oviduct). During incubation, the changes in sperm functional attributes were studied. Sperm kinematics and head area decreased significantly (p < 0.05) immediately upon exposure to hyperosmotic stress at both pH. Proportion of sperm capacitated (%) in 355 mOsm/kg at 1 and 2 h of incubation were significantly (p < 0.05) higher than those in 290 mOsm media. The magnitude and duration of recovery of sperm progressive motility in 355 mOsm with pH 7.4 was correlated with the ejaculate rejection rate (R2 = 0.7). Using this information, the bulls were divided into good (n = 5) and poor (n = 5) osmo-adapters. The osmo-responsive genes such as NFAT5, HSP90AB1, SLC9C1, ADAM1B and GAPDH were upregulated (p < 0.05) in the sperm of good osmo-adapters. The study suggests that sperm are prepared for the osmotic and pH challenges in the female reproductive tract and the osmoadaptive ability is associated with ejaculate quality in bulls.

Centrifugal force assessment in ram sperm: identifying species-specific impact

Background Centrifugation is routinely employed in handling the ejaculates of some species, but it is not part of the commonly used protocols in ram. However, the development and implementation of new assisted reproductive technologies, alternative preservation models based on washing sperm from a cellular ageing-accelerating substance such as the seminal plasma, and basic studies in spermatology is associated with the use of centrifugation. This requires a specific evaluation of the centrifugation protocols considering the species-specific relationship with the potential damage produced by this procedure. No previous studies have determined the effect of different centrifugation forces on ram sperm. Therefore, we aimed to assess the performance of three centrifugal forces (600×g, 3000×g, and 6000×g for 10 min at room temperature) and their effects on ram sperm motility and functionality. Results Sperm motility and functionality parameters were assessed at 0 h and after 2 h of incubation at 37 °C. As expected, a higher cell packaging degree was obtained at high centrifugation forces (P ≤ 0.0001). Cell packaging was unstable at all centrifugal forces. Thus, there was a high cell resuspension rate after less than 2 min. Regarding sperm quality, there was a change in movement pattern of 3000×g and 6000×g centrifuged sperm after 2 h of incubation at 37 °C, characterized by an increase in rapid progressive motility, linearity, straightness, and beat frequency, and a decrease in medium progressive motility, curvilinear velocity, path velocity, and head lateral amplitude. Non-significant differences were obtained among the different treatments concerning the total viability. However, we observed a significant increase (P ≤ 0.05) in the percentage of viable apoptotic sperm in the samples centrifuged at 6000×g at 0 h. Conclusions Centrifugal forces equal to or greater than 3000×g induced some deleterious effects in ram sperm quality, and lower forces did not provide a successful cell packaging degree.

Systematic Analysis of Breed, Methodological, and Geographical Impact on Equine Sperm Progressive Motility

Over the past five decades, there has been increasing evidence to indicate global declines in human semen quality. Parallel adverse trends measured in male animals indicate a potential environmental aetiology. This study evaluated the progressive motility (PM) of stallion ejaculate through a systematic review and meta-analysis. A total of 696 estimates of equine PM from 280 studies, which collected semen samples between the years 1990 and 2018, were collated for meta-analysis. The method of motility analysis, breed, season of collection, and geographical location were extracted. Simple linear regression determined temporal trends in stallion PM. Studies using microscopy estimated PM to be significantly greater compared to computer-automated methods (p ≤ 0.001). For Arabian breeds, PM was consistently higher than other breeds. Over time, there was a significant decline in PM for studies from Europe (n = 267) but a significant increase for studies from North America (n = 259). Temporal trends indicate the fertilising capacity of equine ejaculate has remained consistently high in the last three decades. That being so, variations observed suggest methodological, geographical, and individual stallion differences may significantly influence actual and reported stallion fertility potential.