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Prolonged maintenance of stallion semen by optimization of cooling conditions

JOURNAL OF EQUINE VETERINARY SCIENCE 2025年 144卷 105243页

作者： Guertin, Jillian E. Losano, Joao de Agostini Salazar, Sophia Callaham, Justin Daigneault, Bradford W. Univ Florida Dept Anim Sci 2250 Shealy Dr Gainesville FL 32611 USA

Bottlenecks to the success of equine assisted reproductive technologies (ART) include suboptimal conditions for prolonged storage of stallion sperm. Shipped stallion sperm are transported in cooling devices designed to maintain temperature for up to 48 h. Increasing the storage time of cooled semen while maintaining acceptable motility would relieve logistical ART challenges. Experiments were conducted to test the hypothesis that external regulation of shipment container temperature would prolong storage time of cooled stallion semen. Initial experiments determined the effect of pre-freezing cooling cans at -20 degrees C or -80 degrees C on sperm motility. Fresh sperm was extended in INRA96 and placed in commercial Equitainers for 3.5 days (84 h). Quantification of sperm kinematics was determined every 12 h. Sperm held in Equitainers with -20 degrees C cans maintained higher total and progressive motility than -80 degrees C conditions at 60 h (63 %, 29% vs. 32%, 17%, respectively). Internal monitoring of Equitainers containing -20 degrees C freezer can temperature identified 20 degrees C as the threshold for rapidly decreased motility. In the second experiment, sperm were maintained in Equitainers containing -20 degrees C freezer cans and placed in two different conditions: 1) ambient temperature for 48 h (23 degrees C) or 2) 5 degrees C for external temperature regulation up to 8 days, or when total motility dropped below 50 %. Sperm kinematics was determined every 24 h. Total motility from samples held with external temperature regulation (5 degrees C) remained above 50 % for more than 7 days. These findings suggest that cooled stallion semen can be advanced beyond traditional 48 h constraints by improving temperature maintenance of storage conditions.

关键词： Stallion Cooled semen Motility Temperature

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DNA methylation-based semen age prediction using the markers identified in Koreans and Europeans

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FORENSIC SCIENCE INTERNATIONAL-GENETICS 2025年 77卷 103243-103243页

作者： Lee, Ji Eun Cho, Sohee So, Moon Hyun Lee, Hwan Young Seoul Natl Univ Coll Med Dept Forens Med 103 Daehak Ro Seoul 03080 South Korea Seoul Natl Univ Coll Med Inst Forens & Anthropol Sci Seoul South Korea

In the forensic field, sexual assaults have consistently been the important issue, with semen frequently serving as the primary evidence. When the suspect is unidentified, estimating the perpetrator's age using investigating semen can provide important information. The VISAGE consortium conducted research on the semen age prediction focused on European semen samples, but the age prediction model has remained undisclosed. Additionally, several studies have reported methylation differences across populations, indicating that the European semen age prediction model might not be broadly applicable to other groups. A study did explore semen age prediction in Koreans using Illumina's Infinium Methylation450K BeadChip array, however recent developments in technology could enhance this approach. To address this, we conducted a study on Korean males aged 18-70 years. We initially analyzed 49 samples utilizing Illumina's Infinium MethylationEPIC BeadChip array to identify age-related CpG sites. From this analysis, we identified 9 age-related CpG markers, excluding one due to difficulties in locus-specific analysis. As a result, we used 11 markers including 8 newly identified CpGs from the EPIC array and 3 CpG markers from previous research utilizing the SNaPshot assay. Furthermore, we incorporated 13 CpG markers from the European study to analyze a total of 159 semen samples using the Illumina Nextera MPS system. This approach enabled us to test age-related markers identified in Europeans within the Korean population and to construct a more accurate age prediction model using markers from both Korean and European sources.

关键词： Forensics DNA methylation Age Massively parallel sequencing semen

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Ureaplasma urealyticum upregulates seminal fluid leukocytes and lowers human semen quality: a systematic review and meta-analysis

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BASIC AND CLINICAL ANDROLOGY 2025年第1期35卷 1-22页

作者： Kukoyi, Omotola SeunFunmi Ashonibare, Victory Jesutoyosi Adegbola, Cecilia Adedeji Akhigbe, Tunmise Maryanne Akhigbe, Roland Eghoghosoa Univ klinikum Aachen Funct Microbiome Grp Uniklin Aachen Aachen Germany Oasis Grace Hosp Reprod Biol & Toxicol Res Lab Osogbo Osun State Nigeria Heinrich Heine Univ Med Fac Dept Cardiovasc Surg & Res Grp Expt Surg Cardiovasc Regenerat Med & Tissue Engn CURE 3D Lab Dusseldorf Germany Ladoke Akintola Univ Technol Dept Physiol Ogbomosho Oyo State Nigeria Osun State Univ Dept Agron Ejigbo Campus Ejigbo Osun State Nigeria

Background Ureaplasma urealyticum belongs to the class Mollicutes and causes non-gonococcal urethritis, an inflammation of the urethra that is linked with impaired semen quality. However, some reports are contradictory, and the reported effect of U. urealyticum on specific sperm variables is not consistent. Thus, this study synthesized findings from published primary data and provides a robust and reliable inference on the impact and associated mechanisms of U. urealyticum on sperm quality. Methods A systematic search was conducted until 31st May, 2024, on Cochrane, Google Scholar, and Pubmed. The Population, Exposure, Comparator/Comparison, Outcomes, and Study Design (PECOS) model was adopted. The populations were male in their reproductive ages who were infected with Ureaplasma urealyticum and confirmed positive versus the control who were age-matched non-infected or treated, while the outcomes included conventional semen parameters, seminal fluid leucocyte count, and sperm interleukin-6 (IL-6) concentrations, and the studies were either cross-sectional or longitudinal. Results When compared with the control, quantitative analysis demonstrated that U. urealyticum significantly reduced ejaculate volume (SMD 0.33 [95% CI: 0.15, 0.52] p = 0.0004), sperm concentration (SMD 0.47 [95% CI: 0.31, 0.64] p < 0.00001), total sperm motility (SMD 0.73 [95% CI: 0.43, 1.02] p < 0.00001), total motile sperm count (SMD 0.21 [95% CI: 0.17, 0.26] p < 0.00001), normal sperm morphology (SMD 0.88 [95% CI: 0.42, 1.35] p = 0.0002), but increased seminal fluid leukocyte count (SMD -0.82 [95% CI: -1.61, -0.02] p = 0.04). In addition, qualitative analysis revealed that U. urealyticum-positive subjects had significantly higher levels of IL-1 beta, IL-6, IL-8, TNF-alpha, peroxidase, leukocytes, neutrophils, CD4 + T cells, and CD8 + T cells in the seminal fluid when compared with the control. Furthermore, higher sperm DNA fragmentation and apoptotic sperm cells were observed in U. urealyticum-

关键词： Immunology Infection Male infertility semen Testosterone Ureaplasma urealyticum Ureaplasma urealyticum

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Influence of tannin-rich extract from commercial Acacia mearnsii and gallocatechin on ovine cryopreserved semen viability

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SYSTEMS BIOLOGY IN REPRODUCTIVE MEDICINE 2025年第1期71卷 90-101页

作者： Liman, Mohammed S. Hassen, Abubeker Smuts, Mario P. Biraima, Ahmed D. A. Sutovsky, Peter Mcgaw, Lyndy J. Holm, Dietmar E. Univ Pretoria Fac Vet Sci Dept Prod Anim Studies Pretoria South Africa Minist Livestock & Fisheries Dept Range Management & Grazing Reserve Niger State Livestock & Fisheries Inst Minna Nigeria Univ Pretoria Fac Nat & Agr Sci Dept Anim & Wildlife Sci Pretoria South Africa Univ Khartoum Fac Anim Prod Dept Meat Prod Khartoum Sudan Univ Missouri Div Anim Sci Columbia MO USA Univ Missouri Dept Obstet Gynecol & Womens Hlth Columbia MO USA Univ Pretoria Fac Vet Sci Dept Paraclin Sci Phytomedicine Programme Pretoria South Africa

The objective of this study was to evaluate the effect of a tannin-rich extract from commercial Acacia mearnsii (MTE_0), and gallocatechin, a flavonoid compound derived from Acacia mearnsii, on the long-term viability and motility of cryopreserved ovine semen. Six fresh ejaculates obtained from six adult merino rams twice per week for three weeks were allocated to five aliquots (0, 12.5, 25, 50, and 100 mu M gallocatechin added into the Optidyl (TM) extender) before cooling and cryopreservation. Effects of MTE_0 and gallocatechin on post-thawed motility characteristics were analyzed using computer-assisted semen analysis (CASA), and viability (LIVE/DEAD (R) kit, Molecular Invitrogen, Waltham, MA), oxidative stress (2,7-dichlorodihydrofluorescein diacetate (H2DCFDA, Thermo Fisher (R), Waltham, MA)) for reactive oxygen species (ROS), mitochondrial membrane potential (JC-1 MitoTracker, Molecular Invitrogen, Waltham, MA), acrosomal integrity (lectin PNA), and capacitation using merocyanine 540 (M540) and YO-PRO-1 dyes in flow cytometry. Data were analyzed using one-way ANOVA (IBM SPSS 21.0 for Windows, Armonk, NY). Gallocatechin at 25 mu M positively affected (p <= .001) kinematic parameters including average path velocity (VAP), progressive velocity (VSL), and beat cross frequency (BCF) of cryopreserved semen. Similarly, gallocatechin at 25 mu M- improved sperm motility (live 21.99 +/- 2.06%), reduced ROS levels (26.45 +/- 1.10%), and mitigated premature capacitation (viable and stable 20.08 +/- 1.48%) compared to other treatments. Gallocatechin addition to semen resulted in a significant (p <= .001) positive effect compared with the MTE_0 extract. It is concluded that gallocatechin inclusion at 25 mu M significantly reduces semen deterioration following cryopreservation. This study is the first to introduce gallocatechin as an efficient antioxidant additive to ovine semen to improve its quality during storage. Our findings will help improve post-thaw ovine semen quali

关键词： Gallocatechin semen cryopreservation CASA flow cytometry

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Mild coronavirus disease 2019: Results of a multicentric prospective study on severe acute respiratory syndrome coronavirus 2 presence in semen fractions and cells and its effects on semen parameters

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ANDROLOGY 2025年第3期13卷 544-554页

作者： Bujan, Louis Mendeluk, Gabriela Mahe, Dominique Llabador, Marie-Astrid Bettiol, Celia Plotton, Ingrid Mansuy, Jean-Michel Delobel, Pierre Daudin, Myriam Moinard, Nathalie Ariagno, Julia Chenlo, Patricia Alvarez, Muriel Dejucq-Rainsford, Nathalie Pasquier, Christophe Hop Paule Viguier Inst Natl Sante & Rech Med Inserm Dev Embryonnaire Fertilite Environnem Serv Med ReprodDEFEINSERM1203 Toulouse France Univ Buenos Aires Clin Hosp Jose San Martin Hosp Clin Jose de San Martin Lab Male FertilINFIBIOCFac Pharm & Biochem Av Cordoba 2351 RA-1120 Buenos Aires DF Argentina Univ Rennes Inst Rech Sante Environnem & Travail Irset Physiol & Physiopathol Tractus Urogenital Urgent INSERM Rennes France Hop Bichat Claude Bernard AP HP Nord Serv Biol Reprod Paris France Hosp Civils Lyon Lab Biochim & Biol Mol Hormonol & Endocrinol Mol Bron France CHU Toulouse Inst Federat Biol Lab Virol Toulouse France CHU Toulouse Hop Purpan Serv Malad Infect & Trop Toulouse France

BackgroundSevere 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 *** 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 *** and methodsProspective 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 ***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 sp

关键词： COVID-19 male genital tract male reproductive function reproductive biology SARS-CoV-2 semen seminal cells sexual transmission risk shedding virus semen

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Sperm Motility Evaluation in Stallion Fresh, Cooled and Frozen semen Using a Portable Computer-Assisted Sperm Analysis System

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REPRODUCTION IN DOMESTIC ANIMALS 2025年第3期60卷 e70052页

作者： Brito, Leonardo F. C. Univ Penn New Bolton Ctr Dept Clin Studies Sch Vet Med Kennett PA 19348 USA

semen analysis is an important laboratory diagnostic test for stallions. Evaluation of sperm motility is integral to basic semen analysis and results are important for breeding management and clinical practice. Computer-assisted sperm analysis (CASA) allows objective sperm motility evaluation and increases analytical precision. The objective of the present study was to validate a portable CASA system (AndroScope). Fresh/cooled semen samples (33 ejaculates, 18 stallions) and frozen semen (40 ejaculates and one epididymal flush, 27 stallions) were evaluated using the AndroScope and the IVOS II CASA system ('gold standard'). Positive associations (R-2 = 0.31 to 0.83, p < 0.0001) between the systems were observed for all sperm motility estimates. The percentage of motile sperm was lower (p < 0.0001) with the AndroScope than with IVOS II (mean difference: -4.2%), but the percentage of progressively normal sperm did not differ between the two systems. Systemic differences were observed for sperm kinetic estimates. Sperm velocities, ALH, and BCF were lower (p < 0.0001) with the AndroScope, observations likely associated with the slower video acquisition frame rate used by this system when compared with the IVOS II (45 Hz vs. 60 Hz, respectively). In conclusion, the AndroScope produced precise results for all sperm motility parameters. The ability to identify and track sperm, and to analyse single sperm kinematics seemed similar to that of the 'gold standard' system used for validation. The AndroScope video acquisition frame rate must be taken into consideration when evaluating sperm kinetics and setting thresholds for classification of different motile sperm subpopulations.

关键词： CASA computer-assisted sperm analysis fertility semen sperm motility stallion

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The assessment of oxidative stress in human semen: chaos and confusion in pursuit of diagnostic precision

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REPRODUCTIVE BIOMEDICINE ONLINE 2025年第2期50卷 104488-104488页

作者： Aitken, Robert John Gharagozloo, Parviz Univ Newcastle Coll Engn Sci & Environm Prior Res Ctr Reprod Sci Discipline Biol Sci Sch Environm & Life Sci Callaghan NSW Australia Hunter Med Res Inst New Lambton Hts NSW Australia CellOxess Biotechnol Res & Dev Ewing NJ USA

The importance of oxidative stress in the aetiology of male infertility has occasioned numerous clinical trials designed to assess the potential of antioxidants for treating this condition. These trials have not returned definitive results, probably because they have never selected participants on the basis of oxidative stress. Clearly, if a moderate to severe state of oxidative stress does not exist in semen, antioxidants can hardly be expected to improve fertility. To resolve this issue, robust, user-friendly point-of-care assays need to be developed that will enable clinicians to quickly diagnose and monitor oxidative stress in patients' semen. Traditional assays of total antioxidant capacity do not fulfil this role, because they are time consuming, expensive and laboratory based. The introduction of an alternative electrochemical system (MiOXSYS (R)) was designed to address this problem. This assay records the static oxido-reductive potential of semen and then creates an index by dividing this measurement by the sperm concentration. The creation of such an index is flawed and undermines many of the data generated to date. This commentary explains the nature of this problem and emphasizes the continuing unmet need for effective diagnostic procedures capable of detecting seminal oxidative stress in the patient population.

关键词： Antioxidant. MiOXSYS (R) Oxidative stress semen Total antioxidant capacity

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Improving Ram semen Low Cryotolerance by Replacing the Seminal Plasma With That of High-Cryotolerant Rams or Extender

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VETERINARY MEDICINE AND SCIENCE 2025年第3期11卷 e70381页

作者： Dayanikli, Cemal Bulbul, Bulent Dogan, Sukru Sengul, Ebru Kirbas, Mesut Kal, Yavuz Ataman, Mehmet Bozkurt Sheep Breeding Res Inst Dept Breeding Tech Balikesir Turkiye Dokuz Eylul Univ Fac Vet Med Dept Reprod & Artificial Inseminat Izmir Turkiye Bahri Dagdas Int Agr Res Inst Dept Anim Breeding Konya Turkiye Selcuk Univ Fac Vet Med Dept Reprod & Artificial Inseminat Konya Turkiye

Ram semen cryotolerance problem continues despite intensive research. This study aimed to evaluate the effect of replacing total seminal plasma (SP) with that of high-cryotolerant rams or extender on semen quality parameters in low-cryotolerant rams. Rams previously determined as low (n = 6) and high-cryotolerant (n = 5) were used in the breeding season. High-cryotolerant rams were only included with their SP in the SP+ group. Each normospermic fresh ejaculate from low-cryotolerant rams was split into three aliquots to form the groups. The control aliquot was diluted using a two-step process. Centrifugation removed SP from the second aliquot (SP- group), and the sperm pellet was rediluted with a Tris-based extender using the same amount of SP removed. The third aliquot's SP was removed as in the SP- group, but the remaining sperm pellet was rediluted with fresh SP harvested from high-cryotolerant rams using the same amounts of removed SP in the SP+ group. semen was rediluted with a Tris-based extender in the SP- and SP+ groups using a two-step process as in the control group. Sperm quality was similar between the groups after dilution and equilibration. However, the post-thaw semen quality was higher in the SP- (one parameter) and SP+ (four parameters) groups than in the control group (p < 0.05). Additionally, these parameters were similar in the SP- and SP+ groups. This study showed that, although an adequate extender had positive effects, replacing the whole SP with that of high-cryotolerant rams may better solve the low cryotolerance problem in ram semen.

关键词： cryotolerance extender ram semen seminal plasma

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In vitro effect of platelet-rich plasma (PRP) on morpho-functional characteristics, oxidative stress, apoptosis and CatSper-1 ion channel in short-term stored (5 °C) ram semen

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SMALL RUMINANT RESEARCH 2025年 244卷

作者： Turk, Gaffari Gungor, Ibrahim brahim Halil Arkali, Gozde Cihangiroglu, Aslihan Cakir Badilli, Nida Acisu, Tutku Can Cinkara, Serap Dayan Yuksel, Suheda Kaya, Seyma Ozer Sonmez, Mustafa Gur, Seyfettin Yuce, Abdurrauf Firat Univ Fac Vet Med Dept Reprod & Artificial Inseminat Elazig Turkiye Firat Univ Fac Vet Med Dept Physiol Elazig Turkiye Siirt Univ Fac Vet Med Dept Reprod & Artificial Inseminat Siirt Turkiye

The objective of this study was to examine the impact of platelet-rich plasma (PRP) on the morpho-functional characteristics, oxidative stress, apoptosis and CatSper-1 ion channel in short-term stored ram semen at 5 degrees C. The study was conducted in six independent replicates on pooled semen samples collected by artificial vagina from six sexually active Akkaraman rams. The pooled semen samples were diluted to a concentration of 400 million spermatozoa per milliliter with a tris-based diluent including 6 PRP concentration (1, 2, 4, 8, 10, 20 %) and no PRP (control) at 38 degrees C. Following the reduction of the temperature to 5 degrees C, diluted semen samples were stored for a period of 96 hours. Motility (total, progressive, rapid, medium, low), kinematic (VCL, VSL, VAP, LIN, STR, WOB, ALH, BCF), morphological (plasma membrane integrity, abnormality), flow-cytometric (mitochondrial activity, acrosomal damage, viability, apoptosis), oxidative stress (MDA, GSH, GSH-Px, CAT), and CatSper-1 analyses were performed at both hours zero (initial measurement) and 96. In all groups, a time-dependent deterioration was observed in nearly all parameters at 96 h in comparison to the 0 h. Compared to the control, live spermatozoon rates (8 % and 10 %), GSH-Px (4 %, 8 % and 10 %) and CAT (4 % and 8 %) activities, and CatSper-1 levels at 96 h were higher, while the apoptotic spermatozoon rates (8 % and 10 %) and MDA levels (4 %, 8 % and 10 %) were lower in semen samples containing PRP. However, in semen samples containing 20 % PRP, the apoptotic spermatozoon rates and MDA levels at 96 h were higher, while GSH-Px activity and CatSper-1 levels were lower than in the control. As a result, it was concluded that the addition of 4-10 % PRP to the diluent containing tris+egg yolk has an effective role in preventing oxidative, apoptotic and ion channel (CatSper-1) damages in ram spermatozoa stored for a short time at 5 degrees C, while the concentration of 20 % is harmful.

关键词： Platelet-rich plasma semen Apoptosis CatSper-1 Oxidative stress Ram

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Mitochondria-targeted antioxidant MitoQ improves the quality of low temperature-preserved yak semen via alleviating oxidative stress

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ANIMAL REPRODUCTION SCIENCE 2025年 273卷 107680页

作者： Zhu, Yanjin Yu, Jun Yang, Qinhui Xie, Yumian Li, Xupeng Chen, Zhuo Xiong, Yan Fu, Wei He, Honghong Yin, Shi Lan, Daoliang Li, Jian Xiong, Xianrong Southwest Minzu Univ Key Lab Anim Sci Natl Ethn Affairs Commiss Chengdu 610041 Peoples R China Southwest Minzu Univ Key Lab Qinghai Tibetan Plateau Anim Genet Resourc Minist Educ Chengdu 610041 Peoples R China

Low-temperature preservation of yak semen during transportation and conservation is crucial to accelerate yak breeding. The effects of low-temperature cooling on yak semen quality, however, are poorly understood. This study aimed to determine the dose-dependent effect of mitochondriatargeted antioxidant "MitoQ" on the motility, oxidative status, and mitochondrial function of yak semen during low-temperature preservation. semen samples were collected from six adult healthy Maiwa yaks and preserved at 4 degrees C in semen extender containing 0, 50, 100, 200, and 400 nM MitoQ, respectively. Firstly, the motility, membrane integrity, acrosome integrity, and abnormity index of yak spermatozoa were evaluated to determine the optimal MitoQ concentration. Next, the effect of MitoQ at the optimal concentration on spermatozoa antioxidant capacity, including reactive oxygen species (ROS) and malondialdehyde (MDA) contents, total antioxidant capacity (T-AOC), and superoxide dismutase content (SOD) levels, as well as mitochondrial membrane potential were analyzed. Up to 96 h of low-temperature storage, 200 nM MitoQ showed the most optimal effect on motility, membrane integrity, and acrosome integrity (P < 0.05) but not on sperm morphology (P > 0.05). Also, 200 nM MitoQ markedly reduced yak spermatozoa ROS and MDA contents for up to 48 h of low-temperature storage (P < 0.05). Finally, 200 nM MitoQ significantly improved T-AOC, SOD, and mitochondrial membrane potential for up to 24, 48, and 72 h of low-temperature storage, respectively (P < 0.05). In summary, semen extender supplementation with 200 nM MitoQ is beneficial for low-temperature yak semen preservation via improving the oxidative status.

关键词： Yak semen MitoQ Cold-store Oxidative stress

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