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Background: Due to global warming seasonal heat stress is an increasing problem in temperate zones. Heat stress not only decreases fertility in females, but can also be detrimental to male fertility. Objectives: We studied the effects of natural summer heat stress during spermatogenesis in Holstein bulls on semen quality parameters and on fertilization performance in vitro and possible intergenerational transmission of effects on the next male generation. Materials and Methods: Semen samples from young Holstein breeding bulls, referred to as F0 founders during this study, were collected during summer (F0 "summer" semen) and the following winter (F0 "winter" semen). Parameters such as ejaculate volume, sperm density, motility, thermoresistance, and in vitro blastocyst rates from these F0 semen samples were determined. In addition, after generation of offspring by artificial insemination, semen samples from F1 male offspring were collected and tested for the same quality and performance parameters to capture intergenerational effects. F1 bulls were raised together under identical conditions and semen was collected at about 1 year after birth. Results: The data showed that in vitro blastocyst rates of F0 "summer" semen samples were lower compared with "winter" semen, whereas blastocyst rates of F1 semen samples did not show significant differences. However, whereas F0 semen samples did not indicate significantly different quality parameters we found that motility of F1 semen samples showed significant differences with higher values when collected from bulls generated with F0 "winter" semen. Discussion and Conclusion: From our data, we conclude that (i) natural summer heat stress during spermatogenesis can affect in vitro fertility parameters and (ii) the observed effects on sperm motility of F1 semen samples suggest intergenerational paternal transmission. [ABSTRACT FROM AUTHOR]

Background: Carriers of reciprocal translocations often have more unbalanced spermatozoa and higher DNA fragmentation rates, elevating reproductive risk. The simple swim‐up method (SSUM) can decrease the amount of spermatozoa with abnormal chromatin structure and fragmented DNA, however, it has limited efficacy in eliminating chromosomally unbalanced sperm. Methods: The spermatozoa of eight Robertsonian translocation (Rob) carriers were split into three groups: original raw semen group (control group); SSUM and swimming trapper method group (STM) processed semen samples. After different semen preparation procedures, semen qualities, sperm chromosomal aneuploidy, and sperm fragmented DNA were evaluated. Results: Although spermatozoa with higher motility was obtained by both SSUM and STM, the population of faster forward moving sperm was greater with STM as compared to SSUM. While the rates of DNA fragmentation were statistically much lower in both groups than ejaculated semen sample, our data showed better effect on the decrease of DNA fragmentation index (DFI) after selection by STM for patients who have high DFI (>20%) in neat semen. For all patients, significant decrease in the frequency of chromosomally unbalanced spermatozoa was observed after selection using STM. Although similar trends can be seen in the SSUM group, a significant difference was identified in one patient only. Conclusions: Use of swimming trapper (STM) is superior for enriching high‐motile and genetically competent sperm in comparison with SSUM. [ABSTRACT FROM AUTHOR]

Background: Socioeconomic differences are present within the population of men who experience infertility and seek treatment. Objective: To study the association of socioeconomic status with semen parameters in a group of men using mail‐in semen analyses. Materials and methods: The records of 11,134 men that used mail‐in semen analyses from a fertility company were identified. Their demographic information, semen parameters, and ZIP codes were collected. Area deprivation index (ADI) was used as a proxy for socioeconomic status and was calculated for each individual using their ZIP codes in order to measure their level of socioeconomic deprivation. A higher ADI signifies a more deprived area. The association between ADI and the semen parameters of this group was measured using linear regression analysis adjusted for age. Results: 11,134 men were included in the study with a median age of 35 years (interquartile range (IQR): 32–40) and a median ADI of 83 (IQR: 68–97). The cohort had a median sperm concentration of 31 million/mL (IQR: 14–59), median total sperm count of 123 million (IQR: 57–224), median total motile sperm of 35 million (IQR: 9–95), median total motility of 32% (IQR: 15–52), progressive motility of 22% (IQR: 9–38), and morphology percent normal of 4% (IQR: 2–7). Higher ADI, indicating lower socioeconomic status, was negatively associated with various semen parameters, including sperm concentration, total sperm count, total motile sperm, and total and progressive motility. Discussion and conclusion: Men who live in more deprived areas are more likely to have worse semen parameters. Further research is needed to thoroughly examine the impact of socioeconomic status on male fertility. A comprehensive approach that targets upstream social, economic, and healthcare factors can possibly alleviate the negative association of socioeconomic status with fertility and semen parameters. [ABSTRACT FROM AUTHOR]

Background: Severe acute syndrome coronavirus 2 can invade a variety of tissues, including the testis. Even though this virus is scarcely found in human semen polymerase chain reaction tests, autopsy studies confirm the viral presence in all testicular cell types, including spermatozoa and spermatids. Objective: To investigate whether the severe acute syndrome coronavirus 2 is present inside the spermatozoa of negative polymerase chain reaction‐infected men up to 3 months after hospital discharge. Materials and methods: This cross‐sectional study included 13 confirmed moderate‐to‐severe COVID‐19 patients enrolled 30–90 days after the diagnosis. Semen samples were obtained and examined with real‐time polymerase chain reaction for RNA detection and by transmission electron microscopy. Results: In moderate‐to‐severe clinical scenarios, we identified the severe acute syndrome coronavirus 2 inside spermatozoa in nine of 13 patients up to 90 days after discharge from the hospital. Moreover, some DNA‐based extracellular traps were reported in all studied specimens. Discussion and conclusion: Although severe acute syndrome coronavirus 2 was not present in the infected men's semen, it was intracellularly present in the spermatozoa till 3 months after hospital discharge. The Electron microscopy (EM) findings also suggest that spermatozoa produce nuclear DNA‐based extracellular traps, probably in a cell‐free DNA‐dependent manner, similar to those previously described in the systemic inflammatory response to COVID‐19. In moderate‐to‐severe cases, the blood–testes barrier grants little defence against different pathogenic viruses, including the severe acute syndrome coronavirus 2. The virus could also use the epididymis as a post‐testicular route to bind and fuse to the mature spermatozoon and possibly accomplish the reverse transcription of the single‐stranded viral RNA into proviral DNA. These mechanisms can elicit extracellular cell‐free DNA formation. The potential implications of our findings for assisted conception must be addressed, and the evolutionary history of DNA‐based extracellular traps as preserved ammunition in animals' innate defence might improve our understanding of the severe acute syndrome coronavirus 2 pathophysiology in the testis and spermatozoa. [ABSTRACT FROM AUTHOR]

Background: Dietary factors, including high sugar intake, may have adverse effects on male reproduction. Studies of the association between sugar‐sweetened beverage (SSB) intake and semen quality have reported inconsistent results. Objective: We estimated the effects of SSB consumption on semen quality in a North American preconception cohort study. Methods: We analyzed baseline data from 690 males (n = 1,247 samples) participating in Pregnancy Study Online (PRESTO) during 2015–2022. Participants aged ≥21 years completed a baseline questionnaire on which they reported information about intake of SSBs, including sodas, energy drinks, sports drinks, and fruit juices. After enrollment, we invited U.S. participants to a semen testing substudy, in which they collected and analyzed two samples using an at‐home semen testing kit. We used linear regression models to estimate adjusted percent differences (%D) and 95% confidence intervals (CI) for associations of SSB intake with semen volume, sperm concentration, total sperm count (TSC), motility, and total motile sperm count (TMSC). We used modified Poisson regression models to estimate adjusted risk ratios (RRs) and 95% CIs for the association of SSB intake with World Health Organization semen parameter cut points. Results: Relative to non‐consumers of SSBs, those who consumed ≥7 SSBs/week had lower semen volume (%D = −6, 95% CI: −13, 0), sperm concentration (%D = −22, 95% CI: −38, 0), TSC (%D = −22, 95% CI: −38, −2), motility (%D = −4, 95% CI: −10, 2), and TMSC (%D = −25, 95% CI: −43, −2). High SSB consumers also had greater risks of low sperm concentration (≤16 million/mL; RR = 1.89, 95% CI: 1.11, 3.21), low TSC (≤39 million; RR = 1.75, 95% CI: 0.92, 3.33), low motility (≤42%; RR = 1.23, 95% CI: 0.87, 1.75) and low TMSC (≤21 million; RR = 1.95, 95% CI: 1.12, 3.38). Associations were stronger among participants with body mass index ≥ 25 kg/m2. Conclusion: Greater SSB consumption was associated with reduced semen quality in a North American preconception cohort. [ABSTRACT FROM AUTHOR]

Background: Clinical palpable varicocoeles in conjunction with isolated teratozoospermia are rarely observed. Therefore, the effects of varicocoelectomy on sperm morphology are not clear. The aim of this meta‐analysis is to compile studies that assess the effectiveness of varicocoelectomy in isolated teratozoospermia to reach a more consistent and reliable conclusion. Material‐Methods: The present meta‐analysis was registered to PROSPERO (CRD42023467933). We utilized the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta‐Analyses) guideline to report the outcomes. Articles published before 1 October 2023 were included in the study. The search terms used were teratozoospermia, isolated teratozoospermia, varicocoelectomy for isolated teratozoospermia, and semen analysis after varicocoelectomy in isolated teratozoospermia. Results: We identified 1,013 studies in full publications or abstracts using the methodology and the search terms. Five studies were included for systematic review, while four were included for meta‐analysis. The five studies (10–14) included 348 patients aged 18–44 years. The pooled analysis revealed a significant improvement in sperm morphology in isolated teratozoospermia patients undergoing varicocoelectomy (Q = 199.42, p < 0.0001; I2 = 98.49%). The pooled analysis revealed no significant improvement in sperm concentration in isolated teratozoospermia patients undergoing varicocoelectomy (Q = 5.69, p = 0.058; I2 = 64.85%). Three of the examined studies provided information regarding pregnancy rate and it was high in all studies. According to the Newcastle–Ottowa scale (NOS) assessment, the total quality score of all studies was 7. The funnel plot test demonstrated a visible asymmetry, and Begg and Mazumdar's rank correlation test confirmed the publication bias (p = 0.04). Discussion: Varicocoelectomy can be an effective and reliable treatment option in patients with isolated morphology abnormalities and clinically palpable varicocoele. Conclusion: This meta‐analysis reported that varicocoelectomy may increase pregnancy rates by improving semen parameters in infertile men with isolated teratozoospermia, although this conclusion requires further evidence. [ABSTRACT FROM AUTHOR]

Background: The rate of male infertility has increased in the last decades. One factor that may improve sperm quality and testosterone is fish consumption, which supplies the organism with essential nutrients. Aim: To determine evidence of a relationship between the frequency of fish consumption and sperm quality. Methods: Seventy‐four healthy men, aged 25–45 years, were involved in this observational study. Semen quality was evaluated by spermogram analyses and testosterone was measured in blood. Volunteers were classified as low, moderate, and high fish consumption groups according to the replies indicated in a questionnaire they had to fill out. Results: The moderate fish consumption group showed better semen quality with all spermogram parameters in the reference range and the low fish consumption group manifested the worst semen quality with sperm morphology below the reference range. The low fish consumption group had lower testosterone levels (3.6 ± 0.9 vs. 4.3 ± 1.5; p = 0.04), decreased nonprogressive spermatozoa (11.9 ± 6.2 vs. 15.8 ± 8.2; p = 0.04), and lower semen volume (3.1 ± 2.0 vs. 3.6 ± 1.3; p = 0.06) compared to the moderate fish consumption group. Positive associations were detected between the frequency of fish consumption, the semen volume, and nonprogressive spermatozoa, respectively. Conclusions: Fish consumption at moderate level such as once a week leads to improvement in sperm quality and testosterone levels. [ABSTRACT FROM AUTHOR]

The rising incidence rate of depression presents a substantial threat to human well‐being. Semen Trigonellae (ST), the dried mature seeds of Trigonella foenum‐graecum L., has a long‐standing traditional reputation for alleviating anxiety and hopelessness. However, the anti‐depressant mechanism of ST remains poorly understood. This study aimed to assess ST's anti‐depressant, as well as explore its potential mechanism from a gut microbial aspect. The Kunming mice were challenged by lipopolysaccharide (LPS) to induce depression‐like behavior and then orally administrated with aqueous extract of ST. The behavioral test, and hippocampal and serum biochemical indicators were detected to assess anti‐depressant effect of ST. We utilized full‐length 16S rRNA gene sequencing to investigate how ST influences gut microbiota modulation. Administration of ST mitigated LPS‐induced depression‐like behaviors. ST reversed the decrease in hippocampal 5‐hydroxytryptamine (5‐HT) and brain‐derived neurotrophic factor (BDNF) levels while reducing serum levels of tumor necrosis factor α (TNF‐α), interleukin 1β (IL‐1β), and interleukin 6 (IL‐6). Microbial analysis revealed that administration of ST markedly shifted the gut microbiota structure, dramatically and dose‐dependently increased the abundance of Ligilactobacillus murinus and Ligilactobacillus animalis. Experimentally, oral administration of live L. murinus and L. animalis to LPS‐challenged mice yielded similar effects to ST in ameliorating depression, elevating 5‐HT and BDNF, and reducing proinflammatory cytokines. These findings provide evidence that ST is a promising medical food for the management of depression, in which modulation of the gut microbiota, particularly enhancing the abundance of Ligilactobacillus plays an important role. [ABSTRACT FROM AUTHOR]

The present experiment was carried out to investigate the role of Oxyrase in preserving the in vitro quality, redox status and in vivo fertility of crossbred boar spermatozoa. A total of 24 ejaculates from 6 crossbred (n = 4 from each boar) boars were collected and extended in Beltsville Thawing Solution (BTS) in 1:2 ratio and divided into three aliquots. The first aliquot served as a control (without Oxyrase). Rest of the two aliquots were supplemented with 0.125 (T1) and 0.25 IU/mL Oxyrase (T2). Semen samples were preserved at 15°C for 5 days and kinematics of spermatozoa by CASA, semen quality parameters and oxidative stress status were evaluated at 0, 72 and 120 h of storage. The findings of studies revealed that supplementation of Oxyrase at 0.25 IU/mL resulted in higher (p < 0.05) total motility, progressive motility, plasma membrane integrity, acrosome integrity and functional integrity of plasma membrane at 72 and 120 h in comparison to the control group. Mitochondrial membrane potential (MMP) was higher (p < 0.05) at 72 and 120 h, whereas higher (p < 0.05) DNA integrity was observed at 120 h in T2. The lipid peroxidation (LPO) was lower (p < 0.05) and superoxide dismutase activity (SOD) and total antioxidant capacity (TAC) were higher (p < 0.05) in the T2 group at 120 h as compared to control. In vivo fertility trials indicated a higher (p < 0.05) litter size in T2 in comparison to other groups. The study concluded that the inclusion of Oxyrase at 0.25 IU/mL in the extender protects the crossbred boar spermatozoa against oxidative damage and improves the in vivo fertility. [ABSTRACT FROM AUTHOR]

Premature acrosomal exocytosis in cryopreserved semen is one of the reasons attributed to low fertility among livestock. In the present study, we attempted to enhance the cryopreserved semen quality by selective removal of acrosome‐reacted spermatozoa using FITC‐PNA conjugated iron magnetic nanoparticles (MNPs). Further, the effect of nano purification on other sperm functional attributes was also assessed. Iron MNPs were prepared using co‐precipitation method and dextran‐coated MNPs were conjugated with FITC‐PNA (0.04 mg/mL). A preliminary experiment was conducted to standardise the dose of FITC‐PNA conjugated iron MNPs (0.02, 0.05, 0.1, 0.15, 0.2, 0.4 and 0.6 mg). Among the different doses used, 0.6 mg FITC‐PNA conjugated iron MNPs significantly (p < 0.05) removed higher acrosomal reacted spermatozoa from the semen, and therefore, this dose was used in further experiments. Cryopreserved semen from Holstein Friesian breeding bulls (n = 6) were thawed and washed using Sperm‐TALP to remove residual extender. Washed spermatozoa (2 × 106) were exposed to 0.6 mg of FITC‐PNA conjugated iron MNPs for 10 min at 37°C. The nano purified semen was assessed for various vital sperm parameters viz., viability, intracellular calcium, apoptosis, mitochondrial ROS and mitochondrial membrane potential using flow cytometry. We found that nanopurification using FITC‐PNA conjugated iron MNPs significantly (p < 0.05) improved the sperm quality. The proportion of viable non‐apoptotic spermatozoa with low intracellular calcium levels was significantly (p < 0.05) enriched in nano purified semen. Nano purification did not affect sperm mitochondrial membrane potential and ROS production. In conclusion, these preliminary findings indicate that FITC‐PNA coated iron MNPs effectively removed acrosome reacted spermatozoa and significantly improved sperm functional attributes in the purified fraction. [ABSTRACT FROM AUTHOR]

The main goals of this study were to document and compare the normal ranges of testicular haemodynamic parameters in pre‐ (aged 8–12 months; n = 4) and postpubertal (aged 24–259 months; n = 16) Miranda donkeys in the breeding season, and to correlate animal biometric data and testicular Doppler indices with basic semen quality parameters of sexually mature jacks. Colour and pulsed‐Doppler ultrasonography were employed to assess blood flow in the left and right distal supratesticular artery (DsTA) and their marginal branches (marginal arteries—MA). Peak systolic velocity (PSV), end diastolic velocity (EDV), pulsatility index (PI) and resistive index (RI) were evaluated in both blood vessels, and TAMV (time‐averaged mean velocity), TABF (total arterial blood flow) and TABF rate (TABF‐R) were calculated for MA. The mean diameter of MA was greater (p < 0.05; 0.24 ± 0.05 vs. 0.19 ± 0.05 cm; mean ± SD) but TABF‐R was less (p < 0.05; 0.004 ± 0.004 vs. 0.02 ± 0.01 mL/s/cm3) in sexually mature donkeys compared with prepubertal animals. Apart from RI values for the right testicle of prepubertal donkeys, PI and RI were consistently greater (p < 0.05) in DsTA compared with MA. Significant correlations were found among select biometric and haemodynamic attributes of the testes (height, width and length, TV, TTV and PSV‐ST) and ejaculate characteristics (volume, sperm defects‐total, head and midpiece) in sexually mature donkeys (n = 8). The present results highlight the importance of scrotal ultrasonography for the reproductive assessment of jacks and provide reference values, based on the available subpopulation of Miranda donkeys that can be used in their clinical and reproductive management and research, or conservation programmes. [ABSTRACT FROM AUTHOR]

Sperm cryopreservation in small ruminant is an efficient strategy to distribute spermatozoa for reproductive programmes, but this process reduces the fertility potential of frozen–thawed spermatozoa. The aim of the current research was to evaluate the impact of different concentrations of cysteamine (CYS) in soybean lecithin (SL)‐based medium on postthawed buck semen quality and fertility potential. Semen samples were collected from five bucks, twice a week, then diluted in the SL‐based extender containing different concentrations of CYS as follows: extender containing 0 mM (control, C0), 1 mM (C1), 2 mM (C2), 4 mM (C4) and 8 mM (C8) CYS. Motility characteristics, membrane integrity, abnormal morphology, mitochondrial activity, acrosome integrity, viability, apoptotic‐like changes, lipid peroxidation, DNA fragmentation, ROS concentration, pregnancy rate and kidding rate were evaluated after freeze–thaw process. In results, C1 resulted in greater (p ≤ 0.05) total motility, progressive motility, average path velocity, membrane integrity, mitochondrial activity, acrosome integrity, viability, pregnancy rate and kidding rate compared to the other groups. Furthermore, supplementation of freezing medium with 1 mM of CYS presented lower (p ≤ 0.05) apoptotic‐like changes, lipid peroxidation, DNA fragmentation and ROS concentration compared to the other groups. On the other hand, C8 presented the least (p ≤ 0.05) total motility, progressive motility, average path velocity, membrane integrity, mitochondrial activity, acrosome integrity and viability as well as the highest (p ≤ 0.05) apoptotic‐like changes, lipid peroxidation, DNA fragmentation and ROS concentration compared to the other groups. Therefore, supplementation of freezing medium with 1 mM CYS could be a helpful strategy to protect buck's spermatozoa quality and fertility potential during cryopreservation process. [ABSTRACT FROM AUTHOR]

In this study, we evaluated sheep sperm quality after using Tris‐citrate‐fructose‐based extender with and without egg yolk, a Tris‐citrate without fructose and with egg yolk and the commercial extender Biladyl®, preserving diluted semen at 15 and 23°C for different times (4, 24, 48 and 72 h). The results showed that the diluents with fructose and egg yolk gave the best results of seminal quality. Moreover, the production of ROS was higher for the temperature of 23°C compared to the temperature of 15°C (control). In addition, VCL and the percentage of spermatozoa with intact acrosome decreased with temperatures of 23°C. Finally, a drastic decrease in sperm quality was observed after 24 hours of preservation for most of the parameters evaluated. [ABSTRACT FROM AUTHOR]

The aim of this experiment was to assess the effect of media viscosity on ram sperm motility, kinematics and rheotaxis in vitro by using methylcellulose as a media thickener. Frozen–thawed semen of three rams was thawed and diluted in Tyrode's albumin lactate pyruvate (TALP) media supplemented with 0%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6% and 0.7% w/v of methylcellulose. Sperm motility and kinematic characteristics were analysed using computer‐assisted sperm analysis (CASA). The rheotactic behaviour was assessed in a microfluidic channel, and the number of spermatozoa that passed the 10 mm point of a microfluidic channel over a 2min period against a flow rate of 30 μm/sec was assessed. The use of media with higher viscosity (higher levels of methylcellulose) resulted in significantly lower (p <.05) sperm motility and kinematic parameters. Moreover, higher levels of methylcellulose reduced (p <.05) the number of spermatozoa that exhibited positive rheotaxis. In conclusion, viscosity affected the kinematic properties and rheotactic behaviour of ram sperm. [ABSTRACT FROM AUTHOR]

The present study compares two protocols for the cryopreservation of chicken semen. Both protocols had an initial low cooling rate in the first step, followed by higher cooling rates around ice nucleation (Protocol 1) or following the dissipation of the latent heat of fusion (Protocol 2) in the second step. Semen ejaculates obtained from 12 roosters were diluted with Rootex with 6% dimethylformamide and frozen following either Protocol 1 (from +5°C to −10°C at 5°C/min and from −10°C to −130°C at 60°C/min) or Protocol 2 (from +5°C to −35°C at 7°C/min and from −35°C to −140°C at 60°C/min). Compared with fresh semen, following both protocols, cryopreservation resulted in reduced post‐thaw sperm quality (p <.001). Post‐thaw percentage of sperm with an intact plasma membrane was greater using Protocol 2 than Protocol 1 (p <.05). The results suggest that high cooling rates around the time of ice nucleation are not recommendable. [ABSTRACT FROM AUTHOR]

Goat production is affected by reproductive seasonality. In vitro embryo production (IVEP) could overcome this effect. This study aimed to evaluate the impact of the season of semen collection/freezing on IVEP of prepubertal goat oocytes and on sperm quality and functionality concerning capacitation. Semen from six fertile bucks was collected, pooled and cryopreserved in spring and autumn and used for IVEP of oocytes recovered during the breeding season. Oocytes were IVM in TCM‐199 with hormones, EGF and cysteamine; fertilized and cultured in BO‐IVF and BO‐IVC media (IVF Bioscience, UK). Semen samples were assessed at 0 and 3 h after culture in capacitating (BO‐IVF, CAP) and non‐capacitating conditions for sperm plasma membrane and acrosome integrity, mitochondrial membrane potential (MMP), intracellular calcium and plasma membrane lipid disorder. Blastocyst production was higher with spring sperm compared to autumn (12.0% vs. 2.1%, respectively; p <.05). After CAP, acrosome reaction and intracellular calcium were higher (p <.05) in spring than autumn sperm. No differences were found in other sperm parameters. In conclusion, seasonal variations in the IVEP of prepubertal goats could be linked to differences in sperm ability to undergo in vitro capacitation. [ABSTRACT FROM AUTHOR]

A protocol for conventional in vitro fertilization (IVF) in horses using fresh semen has been described, using a prolonged incubation in FERT‐TALP medium (22 h) at 38.2°C in the presence of penicillamine, hypotaurine and epinephrine (PHE). Our work aimed to develop a protocol that maintains quality parameters in frozen–thawed equine spermatozoa incubated for 22 h in the presence of PHE using different media (FERT‐TALP and INRA96) and incubation temperatures (30 and 38.2°C). Twelve frozen ejaculates from four stallions were thawed and then incubated in either FERT‐TALP or INRA96 with PHE at 30 or 38.2°C for 22 h. Following incubation, total motility (TM), progressive motility (PM), viability and acrosome integrity were evaluated. The results showed that TM was significantly higher (p <.001) at 30°C in both media, while PM was higher for INRA96 at 30°C compared to 38°C (p <.05). Moreover, INRA96 at 30°C exhibited higher sperm viability and acrosome integrity (p <.001) compared to the other experimental groups. These preliminary results suggest that incubating thawed equine spermatozoa at 30°C with PHE in INRA96 successfully maintains motility, viability and acrosome integrity in equine spermatozoa, indicating its potential use for conventional equine IVF. [ABSTRACT FROM AUTHOR]

Myo‐inositol may be efficient to improve sperm parameters to increase the chance of fertility. Although, the data are controversial. This study aimed to assess the impact of Myo‐inositol supplements on semen quality and in vitro fertilization (IVF) outcomes. In this systematic review and meta‐analysis, a comprehensive search was conducted in PubMed, Web of Science, and Embase. The objective was to identify relevant human studies that investigated the effects of Myo‐inositol treatment on various sperm factors, such as sperm motility, sperm concentration, sperm morphology, viable spermatozoa, spermatozoa with DNA fragmentation, and pregnancy rate. Additionally, the testosterone levels of patients with Oligo‐astheno‐teratozoospermia (OAT) after Myo‐inositol application were considered. The findings of 16 selected studies from 2240 citations indicated significant improvements in several parameters of sperm after Myo‐inositol administration. Myo‐inositol treatment was associated with a notable increase in total sperm motility (SMD 0.90; 95% CI: 0.34 to 1.46; I2 = 0%, p = .001) and progressive sperm motility (SMD 1.48; 95% CI: 0.37 to 2.59; I2  = 0%, p = .008). Additionally, there was a significant improvement in testosterone levels (SMD 0.54; 95% CI: 0.34 to 0.73; I2  = 0%, p < .0001). Furthermore, Myo‐inositol therapy demonstrated a significant decrease in spermatozoa with DNA fragmentation (SMD −1.37; 95% CI: −2.43 to −0.32; I2  = 85%, p = .01). This study suggests that Myo‐inositol therapy has a positive impact on specific sperm parameters, such as total and progressive sperm motility, along with testosterone levels. These findings provide support for the potential benefits of Myo‐inositol in improving male fertility parameters related to sperm factors. [ABSTRACT FROM AUTHOR]

The addition of antioxidants to cryopreservation media reportedly improves sperm post‐thaw quality and reproductive performance after artificial insemination. Therefore, the objectives of this study were to evaluate if the addition of L‐carnitine and pyruvate to freezing media, or their addition to samples after thawing, improves the post‐thaw quality of equine spermatozoa. Thus, in Experiment 1, stallion semen samples were cryopreserved in: (1) EDTA–glucose–based extender with 20% egg yolk and 5% dimethylformamide (EDTA control); (2) skim milk–based extender with 20% egg yolk and 5% dimethylformamide (milk control); (3) Extender 1 supplemented with 50 mM L‐carnitine and 10 mM pyruvate (EDTA‐carnitine‐pyruvate); and (4) Extender 2 supplemented with 50 mM L‐carnitine and 10 mM pyruvate (milk‐carnitine‐pyruvate). In Experiment 2, 50 mM L‐carnitine and 10 mM pyruvate were added post‐thaw to samples cryopreserved with extenders 1 and 2 (EDTA control and milk control). Sperm kinematic parameters, DNA fragmentation, membrane lipid peroxidation, acrosome status and viability were evaluated after thawing. No significant differences (p > 0.05) were observed for most of the kinematic parameters, DNA fragmentation, membrane lipid peroxidation, acrosome status and viability of spermatozoa, between the samples frozen in the presence or absence of L‐carnitine and pyruvate, nor between the samples after the post‐thaw addition of these components. A higher (p < 0.05) mean velocity and higher (p < 0.05) amplitude of lateral head displacement were observed in the samples frozen in the milk‐based extender with the addition of L‐carnitine and pyruvate after thawing. The addition of 50 mM L‐carnitine and 10 mM pyruvate, either to the freezing extenders or after thawing, was not deleterious for sperm; however, it did not improve equine sperm motility, viability, acrosome and DNA integrity, nor decrease membrane lipid peroxidation after thawing. [ABSTRACT FROM AUTHOR]

Objectives: This study aimed to investigate the correlation between testicular shear wave elastography (SWE) values and semen analysis results in men with infertility. Methods: This was a retrospective case–control study. Patients were categorized as normal, abnormal, or azoospermic based on sperm analysis results. Testicular volume was measured using B‐mode ultrasonography using the Lambert formula. Subsequently, 40–80 regions of interest measuring 1.5 × 1.5 mm were manually positioned in both testicles based on their size, and two‐dimensional SWE was applied through virtual touch imaging quantification software. Results: The patients had a mean age of 33.79 ± 6.3 years, with semen analysis revealing normal results in 15 patients (22.4%), pathological findings in 35 patients (52.2%), and azoospermia in 17 patients (25.4%). Right, left, total, and mean testicular volumes were significantly lower in patients with azoospermia compared to those in both normal and impaired semen parameters (P <.05). Conversely, testicular elastography scores were higher in patients with azoospermia than in the other groups (P <.05). The significant negative correlation between volume and elastographic findings remained independent of age (r = 0.4, P <.001). The accuracy rates for detecting impaired semen parameters and azoospermia were 94.3% and 94.1%, respectively, after considering factors such as age, testicular volume (right/left/total), and elastography (right/left/total). Notably, the total mean elastography score ranked first, with 100% in the independent normalized importance distribution of these variables. Conclusion: SWE can be used effectively alone or in combination with other diagnostic tools to evaluate histopathological changes in the testicles of male patients with infertility. [ABSTRACT FROM AUTHOR]

Background Objectives Materials and methods Results Discussion and conclusion Severe acute respiratory syndrome coronavirus 2, the virus responsible for coronavirus disease 2019, affects multiple organs. The virus enters cells through angiotensin‐converting enzyme‐2 and host factors present in genital organs, leading to concern over virus shedding in semen and reproductive function.To investigate severe acute respiratory syndrome coronavirus 2 in semen from patients with a mild infection, identify the seminal infected cells, and explore the effect of the infection on sex hormones and semen parameters.Prospective study of 54 men with mild severe acute respiratory syndrome coronavirus 2 infection. Semen was collected at 7, 15, 30, 60, 90, 180, and 365 days after symptom onset, and severe acute respiratory syndrome coronavirus 2 RNA was measured in serum, saliva, urine, and semen. The presence of infectious severe acute respiratory syndrome coronavirus 2 in semen was assessed using Vero cell culture. Infected semen cells were identified using immunofluorescence against severe acute respiratory syndrome coronavirus 2 nucleoprotein antigen and cell markers. Semen characteristics as well as testosterone, inhibin B, luteinizing hormone, and follicle‐stimulating hormone levels were determined.11% of patients had at least one severe acute respiratory syndrome coronavirus 2 RNA‐positive semen. One patient had viral semen shedding up to day 90 after infection onset, with replication‐competent virus isolated from semen and 40% cell fraction at day 7. After sperm preparation, 90% fraction was severe acute respiratory syndrome coronavirus 2 RNA‐positive at days 7 and 15. The swim‐up fraction was positive only on day 7. In semen, nucleoprotein antigen was detected mainly in exfoliated epithelial cells and less frequently in Sertoli cells. Sperm count and motile sperm count were lower at day 30 than at day 7. Round cells in semen were increased during the acute phase. At days 7 and 15, sperm count and motile sperm count were lower in severe acute respiratory syndrome coronavirus 2 RNA‐positive semen compared with negative semen, while semen volume and follicle‐stimulating hormone levels were increased. Long‐term follow‐up shows no evidence of a detrimental effect on hormonal or semen characteristics.11% of patients with mild coronavirus disease 2019 who were not hospitalized had severe acute respiratory syndrome coronavirus 2 excretions in semen, which persisted for up to 90 days in one patient. No germ cells appeared infected by the virus, but the detection of nucleoprotein antigen‐positive epithelial semen cells and Sertoli cells suggests genital tract infection. Albeit infrequent, semen may contain the replication‐competent virus during the acute phase with potential risk of severe acute respiratory syndrome coronavirus 2 transmissions during sexual contact and assisted reproduction procedures. The effect of mild coronavirus disease 2019 on spermatogenesis and reproductive hormones was moderate and reversible. [ABSTRACT FROM AUTHOR]

This study aims to compare the efficacy of computer‐assisted sperm analysis (CASA) and smartphone‐applied sperm analysis (SASA) in assessing the quality of frozen–thawed bull semen. A total of 75 straws (n = 75) semen samples were used from different production batches of five Holstein bulls. The semen analyses were conducted in three groups: Group I (CASA‐37°C), semen samples were evaluated using the CASA system at 37°C (n = 25); Group II (SASA‐25°C), semen samples were assessed using the SASA system at a temperature of room heat (25°C) (n = 25); and Group III (SASA‐37°C), semen samples were evaluated using the SASA system at 37°C (n = 25). The frozen–thawed bull semen samples were analysed in terms of total motility (TM), progressive motility (PM), immotile, velocity average path (VAP), velocity curve linear (VCL), velocity straight line (VSL) and sperm concentration. There was no significant difference between the groups in terms of spermatozoa concentration (p >.05). However, significant differences among the groups were observed for total motile spermatozoa values (p <.001). Values of progressive motile spermatozoa were lower in Group I and Group II compared to Group III (p <.001). The immotile spermatozoa values were significant between the groups (p <.001) and were found to be proportional to total motile spermatozoa values. Additionally, the VAP, VCL and VSL values were comparable between Group II and Group III, but lower when compared to Group I. In conclusion, the results of the study demonstrate that the Sperm Cell™ system can accurately analyse the concentration of frozen–thawed bull semen. The analyses performed at room temperature indicate a parallelism between the PM value and CASA results. However, it is thought that SASA devices require a series of standardization studies in different semen extenders, different sample concentrations and different animal species, analogous to the standardization evolution process of CASA devices in semen analysis. [ABSTRACT FROM AUTHOR]

In vitro capacitation allows for a greater understanding of the mechanisms underlying fertilization and the development of improved reproductive techniques for improving fertility rates in porcine. Tyrodes albumin lactate pyruvate (TALP) and modified Krebs Ringers Broth (m‐KRB) are two medias that are commonly used in research experiments to induce capacitation in boar spermatozoa (Cañón‐Beltrán et al., Theriogenology, 198, 2023 and 231; Oberlender et al., Archivos de Medicina Veterinaria, 44, 2012 and 201; Sahoo et al., International Journal of Biological Macromolecules, 241, 2023 and 124502). Moreover, understanding the morphological and functional changes in boar spermatozoa at different hours of capacitation periods might aid in the development of novel techniques for improving sperm quality and increasing the litter size. This study was carried out to investigate the effect of Tyrode albumin lactate pyruvate and modified Krebs Ringers Broth media on in vitro capacitation of HD‐K75 boar spermatozoa at three different periods of incubation. A total of 24 ejaculate from four clinically healthy, 10–12 months aged HD‐K75 boars, maintained at ICAR‐All India Coordinated Research Project (AICRP) on pig were selected. Semen was collected by 'Simple fist' method using a portable dummy. The semen samples having 200 mL volume, 103 × 106 spermatozoa/ml concentration and 70% initial motility were selected and split into two parts and suspended in TALP and m‐KRB media, respectively, and incubated for 5 h at 37°C. Seminal parameters viz. sperm viability, plasma membrane integrity and acrosomal integrity were estimated in the samples at 0, 3 and 5 h of incubation. This study revealed that there was significant variation between media in live acrosome‐reacted (p <.05) and HOST‐reacted (p <.01) spermatozoa, while between capacitation periods significant (p <.01) variation was observed in hyperactivated spermatozoa, live acrosome‐reacted spermatozoa, HOST‐reacted spermatozoa, FITC‐labelled PSA, extracellular protein and sperm cholesterol. Non‐significant variation was observed in total phospholipid. TALP showed overall better consequence on sperm viability, plasma membrane and acrosomal integrity of boar spermatozoa. From this study, it could be concluded that both TALP and m‐KRB media were virtuous to induce capacitation in HD‐K75 boar spermatozoa. TALP media, however, had a better effect on sperm viability, plasma membrane and acrosomal integrity of boar spermatozoa. Out of the three different periods, 3 h capacitation period resulted in significantly (p <.01) higher incidence of sperm viability, plasma membrane and acrosomal integrity in HD‐K75 boar spermatozoa. [ABSTRACT FROM AUTHOR]

Sepsis causes systemic inflammatory responses and acute lung injury (ALI). Despite modern treatments, sepsis‐related ALI mortality remains high. Aqueous extract of Descuraniae Semen (AEDS) exerts anti‐endoplasmic reticulum (ER) stress, antioxidant and anti‐inflammatory effects. AEDS alleviates inflammation and oedema in ALI. Sodium‐potassium‐chloride co‐transporter isoform 1 (NKCC1) is essential for regulating alveolar fluid and is important in ALI. The NKCC1 activity is regulated by upstream with‐no‐lysine kinase‐4 (WNK4) and STE20/SPS1‐related proline/alanine‐rich kinase (SPAK). This study aimed to investigate the effects of AEDS on lipopolysaccharide (LPS)‐induced ALI model in A549 cells, considering the regulation of ER stress, WNK4‐SPAK‐NKCC1 cascades, inflammation and apoptosis. Cell viability was investigated by the CCK‐8 assay. The expressions of the proteins were assessed by immunoblotting analysis assays. The levels of pro‐inflammatory cytokines were determined by ELISA. The expression of cytoplasmic Ca2+ in A549 cells was determined using Fluo‐4 AM. AEDS attenuates LPS‐induced inflammation, which is associated with increased pro‐inflammatory cytokine expression and activation of the WNK4‐SPAK‐NKCC1 pathway. AEDS inhibits the WNK4‐SPAK‐NKCC1 pathway by regulating of Bcl‐2, IP3R and intracellular Ca2+. WNK4 expression levels are significantly higher in the WNK4‐overexpressed transfected A549 cells and significantly decrease after AEDS treatment. AEDS attenuates LPS‐induced inflammation by inhibiting the WNK4‐SPAK‐NKCC1 cascade. Therefore, AEDS is regarded as a potential therapeutic agent for ALI. [ABSTRACT FROM AUTHOR]

Background: Low sperm motility, one of the common causes of male infertility, is associated with abnormal sperm quality. Currently, important sperm/semen biomarkers are sperm chromatin status and oxidation‒reduction potential (ORP) in semen. Because the association between sperm motility and these biomarkers is still not fully clarified, our study was designed to verify the distribution and risk of sperm DNA fragmentation (SDF) and oxidative stress in semen in asthenozoospermic men. Materials and methods: This study was carried out on discharged sperm cells of asthenozoospermic men (isolated asthenozoospermia or coexisted with reduced sperm number and/or morphology), nonasthenozoospermic men (reduced total sperm count and/or sperm morphology) (experimental groups) and normozoospermic men (proven and presumed fertility) (control group). Basic semen analysis was evaluated according to the 6th edition of the World Health Organization manual guidelines. SDF was assessed using the sperm chromatin dispersion test, while static(s) ORP in semen was measured by means of a MiOXSYS analyser. Results: The men from the asthenozoospermic group had lower basic semen parameters than those from the control and nonasthenozoospermic groups. In men with poor sperm motility SDF and sORP, prevalence and risk for > 20% SDF (high level of DNA damage) and for > 1.37 sORP (oxidative stress) were significantly higher than those of control and nonasthenozoospermic subjects. The risk for sperm DNA damage and oxidative stress in asthenozoospermic men was over 10‐fold higher and almost 6‐fold higher than those in control subjects and almost or over 3‐fold higher than those in nonasthenozoospermic men. Conclusions and discussion: Poor human sperm motility coexisted with low basic sperm quality. Sperm DNA damage and oxidative stress in semen were much more frequent in asthenozoospermia. These abnormalities can decrease the sperm fertilizing capability under both natural and medically assisted reproduction conditions. Thus, in asthenozoospermia, the evaluation of sperm chromatin status and oxidation–reduction potential in semen is justified and inevitable, and the appropriate antioxidant therapy can be suggested. [ABSTRACT FROM AUTHOR]

Background: The implications of SARS‐CoV‐2 infection on male fertility remain largely unknown. Besides their well‐known pro‐ and anti‐inflammatory actions, prostaglandins and polyamines are present in semen, where they play key roles in sperm quality. Objectives: To analyze semen parameters, oxidative profile and the seminal fluid prostaglandin and polyamine systems in samples collected from individuals without coronavirus disease 2019 diagnosis and men who recovered from coronavirus disease 2019. Materials and methods: This study compared semen collected from men without positive coronavirus disease 2019 diagnosis with samples obtained from individuals 1–6 months and 7–30 months post SARS‐CoV‐2 infection. Semen parameters, thiobarbituric acid reactive substances, cyclooxygenase 2 expression by fluorescence immunocytochemistry and immunoblotting, prostaglandin levels by enzyme immunoassay, ornithine decarboxylase activity by a radioactive assay, and polyamine and acetylated polyamine levels by thin‐layer chromatography were assessed. Results: In both groups of semen samples from coronavirus disease 2019 recovered men, sperm vitality, total and progressive sperm motility, and putrescine levels were significantly decreased when compared with samples from the uninfected group. In contrast, lipid peroxidation, leukocyte‐associated cyclooxygenase 2 expression, and prostaglandin D2 levels were higher in semen from coronavirus disease 2019 recovered men than in samples from uninfected individuals. While sperm concentration and morphology, ornithine decarboxylase activity, and N‐acetylputrescine levels were statistically diminished in semen obtained up to 6 months after coronavirus disease 2019 recovery, these parameters remained unchanged when samples were collected 7–30 months after coronavirus disease 2019 recovery. Coronavirus disease 2019 vaccination did not show negative effects on any of the parameters evaluated. Discussion and conclusion: Our work provides insights into the detrimental impact of coronavirus disease 2019 on several sperm parameters, in some cases, even more than a year after SARS‐CoV‐2 infection, which would be accompanied by alterations in the seminal fluid prostaglandin and polyamine profiles. Therefore, future treatments targeting the prostaglandin and polyamine pathways in coronavirus disease 2019 recovered men could lead to a successful reinstatement of semen parameters. [ABSTRACT FROM AUTHOR]

The potential evidential value of male underwear in cases of alleged sexual assault is often overlooked. Male underwear can be a critical item in the investigation of alleged sexual assaults. Body fluids/DNA, which may transfer to the penis during sexual contact, may in turn transfer to the inside front of the underwear, and persist for months or years, provided the underwear are not washed. Here, we demonstrate how the case circumstances drive the sampling strategy of male underwear, in order to maximize the effectiveness of the forensic analysis. Sampling considerations including recovery methods and sampling sequence are discussed, and a methodical examination strategy of male underwear is proposed. To highlight the pertinence of male underwear to the investigation of alleged sexual assaults, three real‐life cases are discussed, in which male underwear were examined for multiple body fluids/DNA, and the findings obtained proved evidentially significant. The different cases demonstrate the versatility of male underwear examination in situations, where different body fluids and DNA may transfer based on the specific allegation, and emphasize how targeted sampling can allow the scientist to assess the probability of the findings based on two competing propositions. Accurate sampling strategies are imperative for robust probability assignment in evaluative reporting of scientific findings. [ABSTRACT FROM AUTHOR]

Summary: The use of cryopreserved stallion semen is a significant aspect of equine reproduction, despite the tendency for lower per cycle pregnancy rates than cooled or fresh semen. This decreased fertility of thawed frozen semen is due in part to the innate subfertility of the chosen male breeding population, but also to the direct insults to the sperm cell during the freezing and subsequent thawing process. The mare aspect of frozen semen breeding is intensively managed to compensate for the aforementioned damages, and acceptable pregnancy rates can be achieved. Here we describe the stallion semen cryopreservation process including semen collection, expected causes of damage in processing, freezing and thawing, mare management and future directions. [ABSTRACT FROM AUTHOR]

Objectives: This study aimed to examine the effects of supplementation of vitamin D to the egg‐yolk extender on characteristics of frozen–thawed ram semen. Methods: Semen samples obtained from adult rams were pooled and divided into five equal volumes. It was reconstituted with extenders containing different concentrations of vitamin D: 0 (control), 12.5 (VITD 12.5), 25 (VITD 25), 50 (VITD 50), and 100 ng/mL (VITD 100), and then they were frozen. Sperm motility parameters, plasma membrane functional integrity, acrosomal integrity, DNA fragmentation, and mitochondrial membrane potential of the groups were evaluated after sperm thawing. Results: Total motility and progressive motility were higher in VITD 50 than in all other groups (p < 0.05). Higher sperm straightness, linearity, and wooble were higher in VITD 50 than in the control group (p < 0.05). A similar pattern of VITD 50 was observed for plasma membrane integrity and mitochondrial membrane potential (p > 0.05). Conclusions: In the study, it was observed that adding vitamin D to the extender had a beneficial effect on ram spermatological parameters. In addition, it was concluded that the use of the 50 ng/mL vitamin D in the extender provided more effective protection than the other doses. [ABSTRACT FROM AUTHOR]

The Bachaur is a mediumized draft purpose breed which has been recognized by ICAR‐National Bureau of Animal Genetic Resources (NBAGR) Karnal, India, and presently is on the verge of extinction. Since there are no data regarding the seminal parameters of this breed, this work was performed to evaluate seminal parameters of freshly ejaculated semen. A total of three healthy breeding Bachaur bulls aged 2.5–5 years were selected for the study which were maintained under identical managemental conditions. Semen parameters of these bulls were studied across 10 ejaculates. The average scrotal circumference and testicular weight of the three bulls were 27.78 ± 1.2 cm and 400.67 ± 26.6 g, respectively. The average overall volume (mL), pH, concentration (million/mL), liveability (%), abnormality (%), HOST (%) and acrosome integrity (%) were 2.20 ± 0.19, 6.86 ± 0.06, 1245.60 ± 23.49, 85.09 ± 0.91, 4.13 ± 0.06, 81.16 ± 1.18 and 83.54 ± 1.32, respectively. The average overall mass motility of three Bachaur bulls was 3.57 ± 0.06 in 0–5 scale and individual motility averaged 84.78 ± 1.70 per cent. The volume of ejaculates in Bachaur bull seemed to be lower as compared to other exotic and Indian breeds. However, the semen parameters with regard to mass motility, liveability, abnormalities, hypo‐osmotic swelling test (HOST) and acrosomal integrity seemed similar to other exotic and Indian breeds. [ABSTRACT FROM AUTHOR]

The scenario of the fertile spermatozoa with high fertilizing capability is basically dependent on gene expression‐based epididymal function. The current investigation aimed to declare the varied expression of different candidate genes (PLA2G4D, LCN15, CLUAP1, SPP1, AQP12B, DEFB110 and ESR1) relevant to spermatozoa features between the different epididymal segments in the mature dromedary camels (n = 30). Scrotal contents were collected post‐slaughtering, during the breeding season and the epididymis was separated from the testicles and divided into three segments (caput, corpus and cauda) based on its morphology and anatomical characteristics. Epididymal spermatozoa were harvested from each epididymal portion and evaluated for motility, count, viability and morphology. Samples were grouped depending on their epididymal sperm cells features into high‐fertile (n = 15) and low‐fertile (n = 15) groups. The gene expression of the candidate genes was defined in the isolated RNA from each epididymal portion tissue. The segmental sperm motion and count were significantly (p <.05 and p <.01) higher in the three epididymal parts of high‐fertile camels than the lower ones. There were some candidate genes markedly up‐regulated in its expression in epididymal head of high‐fertile camels (PLA2G4D and LCN15) and low fertile (CLUAP1), while others in the body region of the high‐fertile group (SPP1, AQP12B and DEFB110). Nevertheless, ER1 did not differ in the expression among the epididymal segments. In conclusion, the variant expression patterns of these epididymal genes in relation to the regional spermatozoa features might suggest important roles of these genes in sperm maturation process in the epididymis and focusing more interest on their potential utility as markers for male camel fertility prediction. [ABSTRACT FROM AUTHOR]

The study investigated if gonad maturation in triploid brown trout, Salmo trutta, was entirely suppressed or only delayed, and if triploids could interbreed with diploid counterparts. Ten percent of the total number of 3‐year‐old triploid S. trutta, 15% of 4‐year‐old fish, and 17% of 5‐year‐old fish produced semen. Three and 4 years old triploid fish did not produce eggs, but 15% of the 5‐year‐old fish did so. The quantity and sperm motility of triploid semen did not differ from diploids, but the sperm concentration was significantly lower. When diploid eggs were fertilized with triploid semen (3n × 2n crosses), the percentage of eyed stage embryos, of hatched larvae, and of normal‐shaped larvae did not differ from the diploid controls. Circa 90% of 3n × 2n crosses had a ploidy level of 2.4n. In the remaining percentage of 3n × 2n crosses, the ploidy level was ≥2n and <2.4n. In sperm competition experiments where diploid eggs were fertilized with a mixture of diploid and triploid semen, 52% of the originating larvae had a ploidy level of 2n, 43% of 2.4n, and 5% of the fish were not exactly classified. From the start of feeding to an age of 248 days, the mortality rate of 3n × 2n interploid crosses and of 2n × 2n controls was similar. The growth of interploid crosses was significantly higher than that of controls. In triploid mature females, the egg mass per kilogram of body weight was significantly lower than in diploids. The mass of the non‐hardened eggs and the percentile weight increase during hardening did not differ from diploid eggs. When triploid eggs were fertilized with diploid semen (2n × 3n crosses), the development rate to normal hatched larvae was less than 10%. All originating larvae had a ploidy level of 3n. From the start of feeding to an age of 248 days, 2n × 3n crosses had a higher mortality rate (15%) than diploid controls (<5%). Growth of this type of interploid crosses was reduced in comparison to controls. Therefore, triploids introduced into natural waters for recreational fisheries or escaping from farms may interbreed with diploid counterparts. This not only alters the genotypes of local populations but also changes the ploidy levels. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the protective effects of nanoparticle selenium (SeNP) and sodium selenite (SS) on preventing oxidative stress during the freezing process of dog semen. A total of six dogs were used in the study. The ejaculate was collected from dogs three times at different times by massage method. A total of 18 ejaculates were used and each ejaculate was divided in five experimental groups. The experimental groups were designed to tris extender containing no antioxidants control, 1 μg/mL SeNP1, 2 μg/mL SeNP2, and 1 μg/mL SS1 and 2 μg/mL SS2. Extended semen were equilibrated for 1 h at 4°C, then frozen in liquid nitrogen vapour and stored in liquid nitrogen (~−196°C). After thawing, semen samples were evaluated in terms of CASA motility and kinematic parameters, spermatozoa plasma membrane integrity and viability (HE Test), spermatozoa morphology (SpermBlue) and DNA fragmentation (GoldCyto). Antioxidant enzyme activity (glutathione peroxidase; GPX, superoxide dismutase; SOD, catalase; CAT) and lipid peroxidation (malondialdehyde; MDA) were evaluated in frozen–thawed dog sperm. When the results were evaluated statistically, the progressive motility, VCL, and VAP kinematic parameters in the SeNP1 group were significantly higher than the control group after thawing (p <.05). The highest ratio of plasma membrane integrity and viable spermatozoa was observed in the SeNP1 group, but there was no statistical difference found between the groups (p >.05). Although the ratio of total morphological abnormality was observed to be lower in all groups to which different selenium forms were added, compared to the control group, no statistical difference was found. Spermatozoa tail abnormality was significantly lower in the SeNP1 group than in the control and SS2 group (p <.05). The lowest ratio of fragmented DNA was observed in the SeNP1 group, but there was no statistical difference was found between the groups (p >.05). Although there was no statistical difference between the groups in the evaluation of sperm antioxidant profile, the highest GPX, SOD and CAT values and the lowest lipid peroxidation values were obtained in the SeNP1 group. As a result, it was determined that 1 μg/mL dose of SeNP added to the tris‐based extender in dog semen was beneficial on spermatological parameters, especially sperm kinematic properties and sperm morphology, and therefore nanoparticle selenium, a nanotechnology product, made a significant contribution to the freezing of dog semen. [ABSTRACT FROM AUTHOR]

Artificial insemination (AI) centres select bulls as calves according to their genetic breeding values and raise them until the first semen collection; yet, a high dropout rate of reared bulls is a problem for AI centres. Potential hormonal indicators of bull sexual maturation (cortisol, dehydroepiandrosterone (DHEA), testosterone, oestradiol, insulin‐like growth factor 1 (IGF‐1)) were observed and evaluated in relation to the performance parameters to perhaps identify candidate biomarkers allowing an early selection of bulls as suitable sires. Blood samples from 102 German Holstein calves at 4 ± 1, 8 ± 1 and 12 ± 2 months of age from six AI centres were analysed using validated immunoassays for cortisol, DHEA, testosterone, oestradiol and IGF‐1. Semen analyses included native and thawed diluted semen. Bulls were classified at the first semen collection into groups with good versus poor performance (GP vs. LP). After 2 years, the subsequent differentiation was done in high (HPP), medium (MPP) and low performance persistency (LPP). Age at first semen collection was an important factor for sperm quality. Cortisol concentrations decreased with age, but the cortisol/DHEA ratio decreased with age only in GP bulls (p <.05). Oestradiol and testosterone concentrations both correlated with libido behaviour (p <.05). Testosterone and IGF‐1 concentrations were higher at the time of first semen collection in GP bulls and increased with age (p <.05). In conclusion, testosterone and IGF‐1 concentrations at first semen collection are associated with performance at first semen collection and future performance persistency, and might be useful early biomarkers for consistent sperm producing bulls on AI centres. [ABSTRACT FROM AUTHOR]

Dimensions of linear type traits facilitate selection of livestock for breeding and rearing. To date, use of linear type traits for selection of breeding bulls is highly concentric to scrotal circumference (SC), with probable overlook to other important traits. Present study reported the importance of various gonadal linear type traits on spermatozoa production, age‐related changes in gonadal linear type traits of bulls and predictive ability of these traits on bulls' reproductive potentials. Among all gonadal traits, testicular density (TD), scrotal volume (SV), paired testicular weight (PWT) and SC were found most important predictor variables in order, which can discriminate between good/poor breeding bulls, that is, produced frozen semen doses (FSD) or not. Dimensions of gonadal traits increased significantly up to 36 months age and thereafter, development became slow and negligible. In contrast, TD decreased by 30%, 51%, 64%, 68% and 71% at 12, 24, 36, 48 and >49 months age, respectively, from its base value at 6 months. Bulls of lower TD (≤0.88 g/cm3) had significantly higher ejaculate volume (+9%), sperm motility, sperm concentration (+100 million/mL) and sperm output (+26%)/ejaculate as compared to bulls of higher TD (>0.88 g/cm3). Discriminant function was developed using TD, SV, PWT and SC to identify bulls of superior reproductive potentials. It was concluded that among the investigated traits, TD was the strongest to discriminate between FSD and Non‐FSD bulls. Therefore, our findings suggested that TD could be more potential trait than SC for dairy bulls' breeding soundness evaluation and assessment of reproductive ability. [ABSTRACT FROM AUTHOR]

A variety of parameters, including liquefaction and semen viscosity, affect the sperm's ability to travel and reach the egg for fertilization and conception. Given that the details behind the viscosity of the semen in male camels have not yet been fully clarified, the purpose of this study was to ascertain how the addition of papain affected the viscosity of fresh diluted camel semen. The study examined semen samples derived from camels that had distinct viscosities. Sperm motility, viability, abnormal sperm percentage, concentration, viscosity, morphometry, acrosome integrity and liquefaction were among the evaluations following 0, 5, 10, 20 or 30 min of incubation at 37°C with papain (0.004 mg/mL, 0.04 mg/mL or 0.4 mg/mL; a semen sample without papain was used as a control). A statistically significant interaction between the effects of papain concentrations and incubation time was found (F = 41.68, p =.0001). Papain concentrations (p =.0001) and incubation times (p =.0001) both had a statistically significant impact on viscosity, according to a simple main effects analysis. A lower viscosity was found (p <.05) at 0.04 mg/mL (0.1 ± 0.0) after 10 min of incubation. A simple main effects analysis showed that papain concentrations and incubation time have a statistically significant effect on sperm motility (p =.0001). At 0.04 mg/mL papain, the sperm motility % was higher (p <.05) after 10 min (64.4 ± 4.8), 20 min (68.4 ± 6.2), and 30 min incubation (72.2 ± 6.6) compared to 0, 5 min (38.3 ± 4.1 and 51.6 ± 5.0, respectively). In conclusion, the fresh diluted camel semen had the lowest viscosity properties after 10 min of incubation with 0.04 mg/mL papain, without compromising sperm motility, viability, acrosome integrity and sperm morphology. [ABSTRACT FROM AUTHOR]

Viruses in human semen may be sexually transmitted via free and cell‐mediated viral infection. The potential effects of semen on the infection and sexual transmission of most viruses in semen remain largely unclear. The present study elucidated the inhibitory effects of human seminal plasma (SP) on Jurkat cell (JC)‐mediated mumps virus (MuV) infection. We demonstrated that MuV efficiently infected JCs and that the JCs infected by MuV (JC‐MuV) mediated MuV infection of HeLa cells. Remarkably, SP was highly cytotoxic to JCs and inhibited JC‐MuV infection of HeLa cells. The cytotoxic factor possessed a molecular weight of less than 3 kDa, whereas that of the viricidal factor was over 100 kDa. The cooperation of cytotoxic and viricidal factors was required for the SP inhibition of JC‐MuV infection, and prostatic fluid (PF) was responsible for both the cytotoxic and viricidal effects of SP. The cytotoxic effects we observed were resistant to the treatment of PF with boiling water, proteinase K, RNase A, and DNase I. Our results provide novel insights into the antiviral properties of SP, which may limit cell‐mediated sexual viral transmission. [ABSTRACT FROM AUTHOR]

Background. Currently, there are few studies on the effects of hyperlipidemia on semen parameters and serum hormones in men. In this study, we divided the study subjects into two groups of normal and hyperlipidemia according to the criteria, and observed the differences between semen parameters and serum reproductive hormones in hyperlipidaemic men and normal men, to explore the potential associations between the indicators. Materials and Methods. Eight hundred eighty five men attending infertility clinics in six hospitals from September 2016 to June 2017 were selected. Their lipid levels, semen parameters, and serum reproductive hormone levels were tested, and a total of 480 men with normal lipids and 405 men with hyperlipidemia were selected according to the criteria, and the relationship between semen quality, serum reproductive hormones, lipids and semen parameters, and serum hormones was statistically analyzed. Results. There was no significant difference in semen parameters between hyperlipidaemic men and normal men (P >0:05), serum testosterone levels were significantly lower in hyperlipidaemic men (P <0:05), and there was a negative correlation between triglycerides (TG) and testosterone in the blood (P <0:05).Conclusion. Hyperlipidemia does not affect male semen parameters, and changes in testosterone in hyperlipidaemic men may be related to triglycerides. [ABSTRACT FROM AUTHOR]

Background: Sperm cryopreservation is an important procedure for oligozoospermic subjects at risk of azoospermia and after surgical recovery of spermatozoa in non‐obstructive azoospermic men. Conventional procedures for sperm cryopreservation might be, however, not suitable for samples with a very low sperm number. Objectives: In this pilot study, we investigated the recoveries of sperm motility and viability in severe oligozoospermic subjects (n = 39) after cryopreservation with a tip‐microVapour Fast Freezing, a procedure previously developed by our group for men with good semen quality. Sperm DNA fragmentation was also evaluated in a second group of oligozoospermic samples (n = 16). Materials and methods: We used a Vapour Fast Freezing procedure using 10 μL tips as carrier, and Test Yolk Buffer as freezing medium (tip‐microVapour Fast Freezing). In a subset of samples (n = 22), we compared recovery of motility and viability as obtained with tip‐microVapour Fast Freezing and with a Vapour Fast Freezing procedure using 500 μL straws. Sperm DNA fragmentation was evaluated by the sperm chromatin dispersion test. Results: We found a recovery rate (median [interquartile range]) of 0.29 (0.13–0.41) for progressive motility, 0.30 (0.21–0.52) for total motility and 0.48 (0.29–0.60) for viability. Interestingly, we observed that samples with the poorest motility were apparently less damaged by freezing/thawing. In a subset of samples (n = 22), we directly compared values of viability, progressive motility and total motility by freezing/thawing with tip‐microVapour Fast Freezing and Vapour Fast Freezing conducted with 500 μL straws. We found much better values of all sperm parameters in samples after freezing/thawing with tip‐microVapour Fast Freezing than with Vapour Fast Freezing in 500 μL straws: that is, progressive motility: 7.00 (3.00–8.50)% versus 2.00 (0.00–4.25)%, p < 0.001; total motility: 12.00 (8.00–16.25)% versus 6.50 (1.00–9.25)%, p < 0.001; viability: 29.75 (23.75–45.25) versus 22.50 (13.75–28.13), p < 0.001, respectively. In the second group of oligozoospermic samples, we found that tip‐microVapour Fast Freezing produced lower levels of sperm DNA fragmentation than straws (33.00 [19.75–36.00]% vs. 36.00 [22.75–41.87]%, p < 0.001). Discussion and conclusion: Tip‐microVapour Fast Freezing appears to be a very promising method to cryopreserve semen samples from severe oligozoospermic patients. [ABSTRACT FROM AUTHOR]

The swine gastrointestinal tract contains a great variety of microbes, forming a complex and dynamic ecosystem. Various internal and external factors (e.g. age, breed and diet) may influence its composition. This study aimed to investigate the gut microbial diversity of German Piétrain boars housed on different deep‐litter bedding materials (regional wood shavings, linen, hemp, spelt husks, and wood shavings) via 16S‐rDNA sequencing. Additionally, short‐chain fatty acids were analysed using gas chromatography. Fresh faecal samples (n = 80) from 40 Piétrain boars were collected twice during the trial. Although it can be assumed that boars ingest bedding orally, no differences in the microbiome composition could be found. The main phyla were Firmicutes and Bacteroides. Acinetobacter was identified as a biomarker for sperm quality differences (total sperm motility) in breeding boars. [ABSTRACT FROM AUTHOR]

This study investigated the effects of storage conditions on the quality of chilled ram semen stored at 4°C for 48 h, comparing aerobic and anaerobic conditions. Ejaculates from INRA180 rams were collected and stored under both conditions, with assessments at 0‐, 24‐, and 48‐h intervals. Various sperm parameters were examined, including motility, velocity, viability, morphology, membrane integrity, and lipid peroxidation. Results showed that storage duration significantly impacted sperm quality, leading to a gradual decline from 0 to 24 h and 24 to 48 h. Notably, after the initial 24 h, progressive motility (PM) and membrane integrity (MI) demonstrated distinct responses to storage conditions. Anaerobic storage consistently improved PM and MI values compared to aerobic storage between 24 and 48 h. Anaerobic conditions also enhanced viability and reduced abnormality at the 48‐h mark. Total motility remained stable throughout storage. Velocity parameters (VCL: curvilinear velocity; VSL: straight velocity and VAP: velocity average path) exhibited differences between the 24‐ and 48‐h intervals, with anaerobic storage resulting in higher VAP and VSL values. Moreover, lipid peroxidation exhibited a progressive increase from 0 to 24 h and 24 to 48 h, independent of storage conditions. Remarkably, anaerobic storage consistently yielded lower lipid peroxidation levels compared to aerobic storage, regardless of storage duration. In conclusion, this study highlights that the anaerobic storage proved advantageous for chilled ram semen quality, particularly after the initial 24 h. [ABSTRACT FROM AUTHOR]

Spermatozoa can experience negative changes when subjected to freezing and thawing, including lowered motility, viability and acrosome response. Herein, the effects of different concentrations of soybean lecithin nanoparticles on cryopreserved Holstein bull semen were examined. Semen was collected, cryopreserved and utilized for sperm kinetic parameter analysis following dilution, equilibration and thawing with 0.5% soybean lecithin (E1), the control extender, and 0.75% (E2), 0.5% (E3), 0.25% (E4) and 0.125% (E5) of lecithin nanoparticles. Results revealed that following dilution, the progressive motility (PM) at E3, E4 and E5 of lecithin nanoparticles was higher (p <.05) than it was for E2. After equilibration, compared to the E1, E2, and E3 values, the PM, vitality, normal morphology, membrane integrity and intact acrosome values at the E5 were consistently greater (p <.05). Comparing the percentages of intact acrosome and membrane integrity at E2 and E3 to E4 and E5, a substantial decrease (p <.05) was seen. Following thawing, the percentage of PM improved at E2 and E5, even though their mean PM values were similar (p >.05) compared to E1, E3 and E4. Vigour and progression parameters of sperm (DAP, DCL, DSL, VAP, VCL, VSL and STR) at E5 were higher (p <.05) than those at E1, E2, E3 and E4. In conclusion, the cryopreserved sperm from Holstein bulls revealed outstanding properties both after equilibration and after thawing with 0.125% lecithin nanoparticles, and they were sensitive to high dosages. [ABSTRACT FROM AUTHOR]

The study investigated midpiece defects in sperm from a 5‐year‐old Brangus bull with a high rate of semen batch rejection, due to morphologically abnormal sperm, with no reduction in sperm kinematics. A comprehensive evaluation was conducted over a 16‐month period, involving 28 ejaculates. Notably, despite the high proportion of midpiece defects (average 37.73%, from 3% to 58%), the study revealed stable sperm production, with no discernible differences in the kinematic data before and after cryopreservation. Electron microscopy identified discontinuities in the mitochondrial sheath, characteristic of midpiece aplasia (MPA). The anomalies were attributed to be of genetic origin, as other predisposing factors were absent. Additionally, the electron microscopy unveiled plasma membrane defects, vacuoles and chromatin decondensation, consistent with previous findings linking acrosome abnormalities with midpiece defects. The findings underscored the necessity of conducting thorough laboratory evaluations before releasing cryopreserved semen for commercialization. Despite substantial morphological alterations, the initial semen evaluation data indicated acceptable levels of sperm kinematics, emphasizing the resilience of sperm production to severe morphological changes. This case report serves as a contribution to the understanding of midpiece defects in bull sperm, emphasizing the need for meticulous evaluation and quality control in semen processing and commercialization. [ABSTRACT FROM AUTHOR]

This study evaluates factors influencing pregnancy rates per artificial insemination (P/AI) and pregnancy loss in Lohi ewes undergoing laparoscopic AI with frozen–thawed semen under sub‐tropical conditions. Data from three experiments comprising ewes (n = 358) of mixed parity (nulliparous; NP and parous; P), various body condition score (BCS) and assigned to long‐term (LTP, 11 days) and short‐term (STP, 5 days) oestrus synchronization regimen across high breeding season (HBS) and low breeding season (LBS) were analysed. Laparoscopic insemination was conducted 54 h post‐sponge removal. Pregnancy diagnosis and loss were evaluated on days 35 and 90 post‐insemination via ultrasonography. Results showed parity significantly influenced P/AI, with nulliparous ewes achieving higher pregnancy ratios than parous ewes (p =.001). BCS significantly influenced P/AI (p <.05), with a quadratic relationship observed between BCS and season (BCS*BCS*Season; p =.07). Progestin treatment did not significantly influence the ratio of pregnant ewes (p =.07). Pregnancy losses were significantly higher during LBS than HBS (p <.05), irrespective of progestin treatment. In conclusion, parity and BCS significantly influenced P/AI, with BCS demonstrating a quadratic association with season. Ewes bred during LBS experienced higher pregnancy losses than HBS, irrespective of progestin treatment. [ABSTRACT FROM AUTHOR]

Cryopreservation is a valuable technique used to assist in the genetic improvement of cultured stocks and provide a continuous supply of good‐quality semen for artificial insemination. Conserving semen by cryopreservation serves several purposes (e.g. artificial reproductive technologies and species conservation) and is also used in the clinical treatment of human infertility. However, the lifespan of cryopreserved semen is influenced by a range of factors, including storage temperature, cooling rate, chemical composition of the extender, the concentration of cryoprotectant, reactive oxygen species, seminal plasma composition and hygienic control. The choice of cryoprotectant is a vital factor underlying the success of animal semen cryopreservation. In this regard, extensive research has been carried out on various cryoprotectants, such as egg yolk, dimethyl sulfoxide, methanol, ethylene glycol and dimethylacetamide. Recent studies have also described the use of a range of new cryoprotectants for cryopreservation, including compounds of plant origin (soy), amino acids, antifreeze proteins, carbohydrates and cyclodextrins. Moreover, semen cryopreservation and storage require the use of liquid nitrogen or ultralow refrigeration methods for both long‐ and short‐term storage. This review summarizes the general methods used for freezing semen and discusses the use of traditional and newly emerging cryoprotectants (permeable and non‐permeable) for the cryopreservation of semen in selected fish and mammalian species. [ABSTRACT FROM AUTHOR]

The aim of this study was to assess the addition of 2% sodium caseinate in a commercial egg yolk‐based medium in frozen ovine semen. Eight Dorper males were used for the study. The ejaculate was divided into two portions and frozen without (G1) or with the addition of 2% sodium caseinate (G2). Kinetic parameters were evaluated using CASA (computer‐assisted sperm analysis), and membrane and acrosome integrity as well as oxidative stress were assessed using flow cytometry. After thawing, a thermoresistance test was conducted at time points T0 and T90. For the fertility test, 100 ewes were inseminated with semen from two rams selected based on in vitro parameters, one with good post‐thaw quality (+70% total motility) and the other with low post‐thaw quality (−55% total motility). For the fertility test, the females were divided into 4 groups for insemination: low‐quality ram without caseinate (GBS = 25) and with caseinate (GBC = 25), and high‐quality ram without caseinate (GAS = 25) and with caseinate (GAC = 25). Regarding the results of sperm kinetics, there was a statistically significant difference in the parameters of average path velocity (VAP) and curvilinear velocity (VCL) between the group frozen with BotuBov and the group with added caseinate. At time point T90, straight‐line velocity maintained a trend (p <.06), with BotuBov® (BB group) being superior to caseinate this time, and in the linearity parameter, caseinate was superior to BotuBov®. Flow cytometry analysis showed no difference between any of the evaluated tests. In the fertility test, there was no statistically significant difference in the pregnancy rate between the BotuBOV® group (23%, 11/48) and the sodium caseinate group (BC group) (33%, 17/52), and no differences were observed in the male versus diluent interaction (p =.70). In conclusion, sodium caseinate supplementation did not influence sperm kinetic parameters and the fertility of sheep. [ABSTRACT FROM AUTHOR]

The effects of an aqueous extract of Scabiosa atropurpurea L. (AES) on the reproduction potential of Queue Fine de l'Ouest rams were evaluated over 9 weeks. Eighteen mature (4–6 years old) rams (52.8 ± 2.6 kg) were divided into three groups. The control (C) group was fed oat hay ad libitum with 700 g of concentrate and the other two groups were fed the same diet supplemented with AES at 1 and 2 mg/kg body weight (AES1 and AES2, respectively). Ram sperm was collected with an artificial vagina (2 × 2 days/week) to evaluate sperm production and quality, antioxidant activity, the adenosine triphosphate (ATP) and calcium concentrations. Sexual behaviour and plasma testosterone concentrations were also investigated. The administration of AES improved sexual behaviour (the duration of contact and the number of lateral approaches). The addition of AES also improved individual spermatozoa motility (C: 71.7% ± 6.3%; AES1: 78.3% ± 4.9%; AES2: 83.8% ± 4.4%), the sperm concentration (C: 5.6 ± 0.36; AES1: 6.4 ± 0.81; AES2: 6.7 ± 0.52 × 109 spermatozoa/mL), the ATP ratio (C: 1 ± 0.08; AES1: 2.1 ± 0.08; AES2: 3.3 ± 0.08) and the calcium concentration (C: 5.6 ± 0.24; AES1: 7.7 ± 0.21; AES2: 8.1 ± 0.24 mmol/L). AES treatment decreased the percentage of abnormal sperm (C: 18.5% ± 1.2%; AES1: 16.2% ± 1.1%; AES2: 14.8% ± 0.94%) and DNA damage (C: 62%; AES1: 27%; AES2: 33%) and was associated with elevated seminal fluid antioxidant activity (C: 22 ± 0.27; AES1: 27.1 ± 1.08 and AES2: 27.5 ± 0.36 mmol Trolox equivalents/L) and plasma testosterone (C: 8.3 ± 0.7; AES1: 11.7 ± 0.4; AES2: 15 ± 0.7 ng/L). In conclusion, our study suggests that S. atropurpurea may be potentially useful to enhance libido and sperm production and quality in ram. [ABSTRACT FROM AUTHOR]

Melatonin is an intracellular antioxidant of sperm membrane that protects the cells from lipid peroxidation. Yet, its role as an antioxidant on semen quality of buffalo bulls is still obscure. The present study was undertaken to assess the effect of exogenous melatonin implant (18 mg/50 kg bodyweight) on post-thaw sperm characteristics, oxidative stress, endocrinological profiles and fertility of buffalo bulls. Six apparently healthy breeding Murrah buffalo bulls were randomly selected at bull farm, Guru Angad Dev Veterinary and Animal Sciences University for the present study and divided into two groups viz. control (n = 3) and melatonin implanted group (n = 3). A total of 120 ejaculates were collected from bulls of both groups (n = 60 each) throughout the study period. Most beneficial effects of melatonin implants were observed during post-implantation period. The percentages of post-thaw sperm total and progressive motility, viability and mitochondrial membrane potential were higher (p < .05) in melatonin implanted buffalo bulls compared to controls during post-implantation period. Following melatonin implantation, MDA production in post-thaw semen was lower (p < .05) in melatonin implanted group than in control group. Plasma melatonin and testosterone concentrations were higher (p < .05) in buffalo bulls implanted with melatonin as compared to their control counterparts. No differences (p > .05) in plasma LH concentrations were observed in both groups. First service pregnancy rate was 43.3% using semen of melatonin implanted bulls and 30.0% with semen of controls (p > .05). Thus, melatonin was able to protect sperm membrane against oxidative damage and improve post-thaw semen quality, thereby resulting in higher fertilizing potential of spermatozoa. [ABSTRACT FROM AUTHOR]