Journal of Equine Veterinary Science最新文献

{"title":"Influence of two extenders used for short-cooled storage on motility of Quarter Horse semen submitted to a test freezing","authors":"M. Nunes ,&nbsp;M.L. Munhoz ,&nbsp;B. Teixeira Marimon ,&nbsp;L.R. Domingues Ramos Filho ,&nbsp;M. Gallelli ,&nbsp;G. Ferrer Carneiro ,&nbsp;M. Miragaya","doi":"10.1016/j.jevs.2024.105310","DOIUrl":"10.1016/j.jevs.2024.105310","url":null,"abstract":"<div><div>Identifying the most appropriate extender for short-cooled storage is essential for maximizing post-thawing results. The objectives of this study were assessing the impact of two extenders routinely used for cooled-storage on post-thaw progressive motility (PM) and identifying the most adequate freezing extender for these samples’ cryopreservation. One ejaculate from each of ten Quarter Horse stallions, routinely used in a reproduction center, was collected. Samples were randomly diluted in Botu-Semen (BS) (G1) (n=5) or EquiPlus (EP) (G2) (n=5) to 50 × 106 cells/ml and maintained at 5°C for 8 h, before being equally split into 9 tubes. They were centrifuged (600g x 20 min) and the pellet was resuspended with one of 9 different freezing extenders (Blue, Green, Orange, Red, Violet, Star, MX3, Gent and Botucrio) to 200 × 106 sperm/ml. Then straws were manually filled and kept at 5°C for 20 min in a refrigerator. Subsequently, they were exposed to nitrogen vapors for 20 min, before immersion in liquid nitrogen. Straws were thawed in a water bath at 37°C for 30 seconds. Evaluation of sperm PM after cooling storage (t0) and after thawing (t1) was performed using the CASA System (AndroVision, Minitube, Germany). A generalized linear model was used, being the response variable, PM, and the fixed effects, cooling extender (two levels: G1 and G2) and freezing extender (9 levels: each freezing extender). To analyze the number of samples with post-thaw PM ≥ 30% according to the cooling extender, a generalized linear model was used too. Differences were considered significant when P values were &lt;0.05. Values are expressed as mean ± SD (Graph Pad, USA). Mean PM before freezing did not show significant differences between G1 and G2 (73.4±14.4% vs 72.8±4.3%, respectively). Post-thaw PM was significantly greater in samples of G2 vs. G1 (28.6±10.2% vs 16.15±10.2%) (P˂0.001). Moreover, 48.8% of samples of G2 showed post-thaw PM˃30%, while only 8.8% of G1 did (P˂0.01). Considering each stallion's performance, 5/5 individuals of G2 reached a post-thaw PM˃30%, while only 2/5 of G1 did. Samples of G2 showed a greater post-thaw PM when using STAR, Orange and Violet, compared to G1 (34.6±6.3% vs. 18.7±11.9%, 23.8±8.2% vs. 11.8±5.8% and 38.3±6.7% vs. 19.1±8.9%, respectively). Post-thaw PM˃30% was attained for 4/5 stallions of G2 whose sample was diluted in STAR and Botucrio, and for 5/5 whose sample was diluted in Violet. Only 1/5 stallions of G1 attained post-thaw PM˃ 30% (using STAR, Botucrio, Green and Violet). No interaction was observed between the cooling extender and the freezing extender. In general, mean PM decreased from t0 (73.1±10.1%) to t1 (22.4±11.9%) (P˂0.001). In conclusion, PM after-thawing was better when samples were stored in EP than in BS. Better results were accomplished when using Violet, Star and Botucrio as freezing extenders.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105310"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386863","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of hay nets on subluxations, pain-pressure thresholds, and cervical range of motion in the axial skeleton of adult horses","authors":"M. DeBoer ,&nbsp;T. Rieck ,&nbsp;L. Johnson ,&nbsp;H. Redenius ,&nbsp;K. Martinson","doi":"10.1016/j.jevs.2024.105256","DOIUrl":"10.1016/j.jevs.2024.105256","url":null,"abstract":"<div><div>Horses have become more intensively managed, including the use of hay nets to increase intake time and reduce hay waste, but their impact on health is not well known. The objectives were to compare changes in subluxations, range of motion, and the pain-pressure threshold in the axial skeleton of horses fed hay with (NET) or without (CON) hay nets. In September 2021, 13 adult horses were blocked by bodyweight (BW) and randomly assigned to the NET or CON treatments for one year. After one year, horses switched treatments and the trial concluded in September 2023. Horses were housed in dry lots with free choice access to round bales with or without hay nets (4.45 cm openings). Chiropractic adjustments occurred in May and August each year to note axial skeleton subluxations. Evaluations occurred in June, July, and August to measure cervical range of motion and mechanical nociceptive thresholds (MNTs) of the jaw, poll, and withers. SAS was used and significance was set at <em>P</em>≤0.05. There was an average 1.6-point increase in cervical subluxations observed in NET horses (<em>P</em>&lt;0.05) while no changes were observed for the TMJ and thoracic or lumbar vertebral regions (<em>P</em>&gt;0.05). Average cervical range of motion was 5 degrees greater in NET horses compared to CON horses for chin to chest and chin to fetlock measurements (<em>P</em>≤0.05). No differences were observed in MNTs (<em>P</em>&gt;0.05). These data suggest that netted round bales may only have minor impacts on the axial skeleton of horses over 12-weeks.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105256"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142791595","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Alcohol-based antisepsis without the use of chlorhexidine for arthroscopy in horses","authors":"Y. Nakamae,&nbsp;Y.A. Elce,&nbsp;M.E. Saab,&nbsp;J T. McClure,&nbsp;A.J. Doyle","doi":"10.1016/j.jevs.2024.105257","DOIUrl":"10.1016/j.jevs.2024.105257","url":null,"abstract":"<div><div>Alcohol-based antisepsis has shown experimentally to be as effective as 4 % chlorhexidine gluconate (CHG) at reducing bacterial counts (colony forming units; CFU) on equine skin. Our objectives were to determine the immediate and post-surgical reduction in CFU/mL on equine skin prepared with CHG-based or 70 % isopropyl alcohol (IPA)-based (without CHG) protocols in a clinical setting with arthroscopic surgery. Our hypotheses were that the log₁₀CFU/mL reduction would not significantly differ between protocols immediately after preparation or at the end of surgery. Six horses underwent a 40 min bilateral tarsocrural joint arthroscopy with each limb randomly assigned to Group A or B. Group A tarsocrural joints underwent a rough scrub using 4 % CHG and a 5 min sterile scrub using 2 % CHG. Group B underwent a rough scrub with neutral soap followed by a 90 s sterile scrub with IPA. Samples were collected before rough scrub (T0), immediately after sterile scrub (T1), and end of surgery (T2). CFU/mL were determined in duplicate and were log-transformed and averaged. ANOVA models compared the immediate reduction (T0-T1) and sustained reduction (T0-T2) between treatment groups. The immediate and sustained log10CFU/mL reduction between groups was not different (<em>P</em> = 0.46, <em>P</em> = 0.42). Both groups achieved at least a 2-log immediate and sustained reduction. Limitations include small population size, short surgical duration, length of follow-up, and researchers were not blinded to treatment during sampling. This study demonstrates efficacy of IPA-based antisepsis, without the need for CHG, and supports further investigation in clinical surgery as an acceptable method of surgical site preparation.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105257"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142872195","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The use of sex-sorted semen in horses","authors":"JC Samper ,&nbsp;JC Hernandez Aviles ,&nbsp;LF Ramirez-Agamez ,&nbsp;CC Love ,&nbsp;C Gonzalez-Marin ,&nbsp;P Fleury ,&nbsp;P Dini ,&nbsp;A De La Fuente ,&nbsp;R Foss ,&nbsp;FL Campos ,&nbsp;PJ Ross","doi":"10.1016/j.jevs.2024.105251","DOIUrl":"10.1016/j.jevs.2024.105251","url":null,"abstract":"<div><div>The application of sex-sorted semen in horses has historically lagged the bovine industry due to differences in the reproductive physiology and grater variability in equine semen quality. Recent advancements, including SuperGen™ sorters and Ultraplus™ semen processing methods developed by STgenetics® (Navasota, Texas), have significantly improved the efficiency of the sex-sorting process and reduced the sperm damage previously reported. To facilitate the broader use sex-sorted semen in the equine industry, it is essential to address challenges such as shipping cooled semen to a central sorting facility, minimizing oxidative stress and DNA fragmentation, and developing effective methods for short-term cooling or long-term cryopreservation of the sex-sorted sperm. In this study, we evaluated the feasibility of shipping cooled (8 °C) equine semen to a central facility for sorting and assessed the quality of sperm post-sorting and preservation. Results from the improved process showed enhancements in sperm motility, morphology, viability and DNA fragmentation following sex-sorting. Although there was a slight, consistent increase in lipid peroxidation of sex-sorted sperm, the quality of sperm over the first 24 h post-sorting was comparable to that of conventional (non-sorted) cool-shipped semen. Fertility rates with fresh semen did not differ between conventional and sex-sorted sperm. When using frozen-thawed sex-sorted sperm for intracytoplasmic sperm injection (ICSI), consistent blastocyst production was achieved, albeit at slightly lower rates than those observed with conventional semen for the same stallions. Stallion-specific variability may account for some of these differences. Overall, sex-sorted semen used for ICSI resulted in a higher number of embryos of the desired sex compared to those obtained using conventional semen.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105251"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142791766","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Protocol modification for frozen semen insemination in mares","authors":"J. Hayna ,&nbsp;A. Ragon ,&nbsp;A. Hubner","doi":"10.1016/j.jevs.2024.105292","DOIUrl":"10.1016/j.jevs.2024.105292","url":null,"abstract":"<div><div>Equine breeding commonly employs artificial insemination with increased demand for cryopreserved semen. Classic management strategies have relied on intensive mare examination, commonly with insemination after ovulation. Previous authors (Squires.et.al. AAEP Proc. 2003;49:353-356) looked to reduce examination frequency utilizing timed insemination. Examination frequency was reduced but a second insemination was used. In an effort to reduce examination frequency and eliminate a second insemination, a protocol modification was made in the timing of inseminations. Mares were part of active breeding programs associated with the practices of two theriogenologists. A total of 67 mares, ranging from 3-22 years of age, yielded 127 total cycles which were inseminated over 3 years. The horses represented a variety of breeds, and all mares were to carry their pregnancies. When transrectal palpation and ultrasound showed the mares had a dilated cervix, uterine edema and an ovarian follicle greater than 35mm in size, ovulation was induced. Induction was designated time 0 hours. Mares were induced with either hCG 3000IU IV, deslorelin acetate 1.7mg IM or histrelin 0.5mg IM. Mares were examined and inseminated according to the protocol chosen, 6 Hours, Timed or Proposed. The 6 Hour method used mare examinations every 6 hours until ovulation was detected and inseminated at that time. Timed used examinations at 24, 30 and 48 hours with insemination at 30 and 48 hours. Proposed used examinations at 24, 30, 33-36 and 48 hours, with insemination at the 33-36 hours. Examination at 48 hours was used to confirm ovulation and proper timing. Nine mares were inseminated on 13 cycles in the Proposed group. Thirty mares were inseminated on 38 cycles for the Timed group. 48 mares were inseminated on 76 cycles for the 6 Hours group. Pregnancy rates per cycle for each group were 69.2%, 44% and 48.6% respectively. Statistically there was no difference between the groups. We feel this demonstrates that the Proposed method is an acceptable method to achieve pregnancy per cycle rates that are comparable with previously described methods. Disadvantages of the study was its retrospective nature and the small sample size of the proposed method. Advantages were the removal of examinations between the hours of 10PM and 8AM and the use of a single dose of frozen semen. Adjusting the timing of ovulation induction to the morning eliminated overnight and early morning exams, which had a direct, positive effect on professional wellbeing which cannot be understated.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105292"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386753","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The protective capacity of different diluents on stallion sperm during storage in a passive refrigeration system “BotuflexⓇ '' for 24 h at 5°C before freezing","authors":"L.P. Zanlorenzi-Silva ,&nbsp;C.P. Freitas-Dell'Aqua ,&nbsp;M.E.A. Baby ,&nbsp;C.M. Trinque ,&nbsp;M.S. Frasson ,&nbsp;L.E.F. Canuto ,&nbsp;M.A. Alvarenga ,&nbsp;J.A. Dell'Aqua JR ,&nbsp;T.M.S. Cavalero ,&nbsp;F.O. Papa","doi":"10.1016/j.jevs.2024.105337","DOIUrl":"10.1016/j.jevs.2024.105337","url":null,"abstract":"<div><div>The refrigeration of semen at 5°C for 24 hours before freezing, using Equitainer^Ⓡ and Max-Semen Express^Ⓡ, resulted in fertility rates similar to conventional methods (Melo.et al. Animal Reproduction. 2006; v.94, p.78-81). The objective of this study was to compare conventional freezing and refrigeration prior to freezing, using different refrigeration diluents in Botuflex-Botupharma^Ⓡ, with the purpose of improving sperm motility, plasma membrane stability, mitochondrial potential, and sperm morphology rates. The study used three ejaculates from four stallions of different breeds with proven fertility, collected using a Botucatu^Ⓡ artificial vagina. After, the sperm parameters were measured and the ejaculates were diluted with Botu-Special^Ⓡ (BSP), Botu-Sêmen^Ⓡ (BSEM), Botu-Sêmen Gold^Ⓡ (BG) and Botu-Turbo^Ⓡ (BT), which were divided into two groups, with “C” being the control frozen according to the conventional methodology with Botucrio^Ⓡ and “R”, refrigerated at 5°C for 24 hours and subsequently frozen by the same methodology. After thawing, kinetics (CASA), plasma membrane stability, mitochondrial potential by flow cytometry and sperm morphology (DIC) were evaluated. Statistical analysis was performed by Kolmogorov-Smirnov, ANOVA and Tukey test, P〈 0.05 was considered. The results of the R BSP (36.9 ± 3.5ab) and R BG (43.5 ± 4.2a) groups showed similar stability to the control C BSP (41.3 ± 3.2ab) and C BG (47.9 ± 3.5a). Regarding mitochondrial potential, R BG (46.9 ± 4.0a) and R BSP (39.5 ± 3.5ab) showed similar values to the controls C BG (50.5 ± 3.3a) and C BSP (44.3 ± 3.1ab), suggesting no negative impact on mitochondrial function. As for total motility, R BG (73.9 ± 3.2a) and R BSP (70.8 ± 4.1ab) were similar to their respective controls C BG (79.2 ± 2.8a) and C BSP (79.6 ± 1.9a). Progressive motility was also preserved in R BG (41.9 ± 3.8a) and R BSP (40.0 ± 4a), when compared to the C BG (43.7 ± 3.6a) and C BSP (45.9 ± 3.4a) groups. In terms of fast sperm, the R BG (62.4 ± 4.7a) and R BSP (57.7 ± 5.4a) groups did not show significant reductions. The experimental groups R BSEM and R BT were not mentioned during the discussion as they did not have satisfactory results. The sperm morphology analysis revealed no statistical differences between the groups analyzed. The conclusion is the R BG and R BSP groups showed significant sperm function preservation similar to the controls. Thus, the Botu-Special^Ⓡ and Botu-Sêmen Gold^Ⓡ media can be used to refrigerate semen at 5°C for 24 hours before freezing. This technique allows veterinarians to optimize their work in the field, without having to freeze semen in establishments where conditions are not suitable for the cryopreservation of equine semen.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105337"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386625","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The semen microbiome influences the mare uterine microbiome","authors":"G. Podico,&nbsp;I.F. Canisso","doi":"10.1016/j.jevs.2024.105314","DOIUrl":"10.1016/j.jevs.2024.105314","url":null,"abstract":"&lt;div&gt;&lt;div&gt;The uterine microbiome has been well characterized in farm animals, including horses, and associations between the semen microbiome and uterine microbiome have been described in humans but unknown in horses. Of interest is that high bacterial richness in semen was linked to dysbiosis in the vaginal microbiome of sub-fertile humans. An extra-species model (donkey-to- and horse-to-mare) was designed to determine the effects of the male seminal microbiome on the mare uterine microbiome. This study hypothesizes that the semen microbiome of the horse differs from the jack's, which could trigger distinct post-breeding changes in the mare uterine microbiome. Twenty-two fertile mares were inseminated with a single fertile donkey jack or horse stallion raw semen (2 billion total sperm) in a cross-over design with a washout cycle in between. A sample of uterine fluid was collected before the insemination and after 6h, 24h, and at the embryo flush 8d post-ovulation. Bacterial DNA was purified from semen and uterine fluid with a commercial kit (QIAGEN, QIamp DNA microbiome kit) and submitted for PCR amplification and sequencing of the full-length 16S region. Taxonomy assignments were done via QIIME2, and R was used for the analyses. The dataset was filtered and agglomerated per rank to the Family level. Alpha and beta diversity indexes were calculated and compared with the Wilcoxon rank-sum test, a linear mixed model, and PERMANOVA; significance was set at P〈 0.05. Alpha diversity (Faith's PD index) differed between donkey and horse semen (P=0.048), whereas beta diversity was similar across species (P 〉0.05). The most prevalent Phyla in mares bred to the donkey were Proteobacteria (50.3%), Actinobacteria (18%), and Firmicutes (29.8%) similarly, in mares bred to the horse, Proteobacteria (62.5%), Actinobacteria (21.7%), and Firmicutes (12.3%) were the most abundant. Noteworthy, the Order of Lactobacillales bacteria contributed up to 20% of the microbiome composition in mares bred to the donkey jack and up to 6% in mares bred to the horse stallion. Twenty-four bacteria families were found to have a common relationship between donkey semen and the mare uterus after the insemination, and nine were identified between horse semen and the uterus. Species richness and evenness (alpha diversity) were not different in mares with a positive or a negative embryo flush (Observed ASVs, P=0.11; Chao1, P=0.22; Shannon, P=0.91; Simpson, P=0.19, Faith's PD, P=0.70). The beta diversity of the uterus 8d post-ovulation differed based on the embryo outcome in both mares bred to the donkey or the horse (R-squared=0.069; F-value=2.01; P=0.02). In conclusion, the semen microbiome varied between the two equids used in the study, and more importantly, the semen microbiome affected the mare uterine microbiome and suggested for the first time in the horse that the presence of a temporary combined male and female microbiome has apparent consequences for the embryonic recovery.&lt;","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105314"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386868","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of the germplasm bank for the Spanish autochthonous breed Asturcon","authors":"C. Tamargo,&nbsp;Á. Fernández,&nbsp;M.J. Merino,&nbsp;C.O. Hidalgo","doi":"10.1016/j.jevs.2024.105325","DOIUrl":"10.1016/j.jevs.2024.105325","url":null,"abstract":"<div><div>Germplasm banks provide an essential service by safeguarding the genetics of endangered or valuable species or individuals and are one of the strategies to preserve endangered domestic breeds (Solti et al. Theriogenology 2000; 53, 149-162). The SERIDA (Asturias, Spain) keeps germplasm from several local breeds. The aim of this work is to characterize the semen of Asturcon ponies, an autochthonous and endangered equine breed adapted to the mountainous Atlantic environment and officially protected (National Official Breed Catalog). The study included 241 ejaculates from 16 pony stallions (aged 6-17 years) collected by artificial vagina, 3 days per week, in breeding season (from April to July) over 5 years (Crespo et al. Animal Reproduction Science 2020; 223,106641). Immediately after collection, gel-free semen was evaluated for: volume, estimated from a graduated tube; sperm concentration, determined with a calibrated spectrophotometer (Accucell, IMV Technologies, L'Aigle, France); and motility, assessed subjectively with a phase contrast microscope (× 4 objective) and heated stage (37°C) and by Computer-Assisted Sperm Analysis (CASA) system (ISAS v. 1.19 software; Proiser, Spain) in fresh and post-thawed semen. Then semen was diluted and centrifuged for 12 min at 600 <em>g</em>, the supernatant was discarded, and the pellet was resuspended in freezing medium (skim milk extender with 2% egg yolk and 2.5% glycerol), to a final concentration of 100 × 10⁶ spermatozoa/ml, and equilibrated/cooled (60 min) to 4°C (Fayrer-Hosken et al. Journal of Equine Veterinary Science 2008; 28, 672-676). Semen doses were packaged in 0.50 mL straws, frozen in a programmable freezer (Digit-cool; IMV Technologies) with a two-steps curve and stored in liquid nitrogen in SERIDA cryobank. Three straws per ejaculate were thawed in a water bath at 37°C for 30 s and pooled before analyses. Statistical analysis was carried out by means of the GLM and CORR procedures and Duncan test for means (SAS Institute, Inc., Cary, NC, USA). A significant effect between males (<em>P &lt; 0.05</em>) on semen quality, such as volumen, sperm concentration and motility, were detected. On the other hand, positive and significant correlations (r=0.76; <em>P &lt; 0.05</em>) were found between sperm motility pre-freezing and post-thawed (total motility 26–43%, progressive 14–28%). It is well known that there are considerable differences between stallions in the success of spermatozoa to survive cryopreservation, and often among ejaculates as well (AI-Kass and Morrell. Veterinary Science 2024; 11, 65). Whereas the sperm parameters of this endangered breed are acceptable, future research should focus on optimizing the existing protocols and on the ability of Asturcon stallions’ sperm to survive freezing and thawing procedures in rates higher than 35% to ensure that germplasm bank are correctly created.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105325"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386701","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Addition of a motility stimulant improves kinetic parameters in post-thawed cholesterol-loaded stallion semen","authors":"V.M.T. Vilela ,&nbsp;B.M. de Zutter ,&nbsp;D.F. Silva ,&nbsp;J.H.W. Diniz ,&nbsp;M.L. Frédou ,&nbsp;V.G. de Araujo ,&nbsp;Y.F.R. Sancler-Silva ,&nbsp;F.S. Ignácio ,&nbsp;F.O. Papa ,&nbsp;G.A. Monteiro","doi":"10.1016/j.jevs.2024.105333","DOIUrl":"10.1016/j.jevs.2024.105333","url":null,"abstract":"<div><div>The freezing process exposes sperm to thermal stress, resulting in structural damage that may compromise their functionality. This study aimed to assess the impact of including cholesterol bound to cyclodextrin (CLC) before cryopreservation and of adding a motility stimulant (FertTalp) (Parrish et al. Theriogenology. 1986; 25:591-600) after thawing, on sperm kinetics, intracellular hydrogen peroxide production, mitochondrial potential and plasma membrane destabilization in equine semen. Twelve Mangalarga Marchador stallions were used to collect two ejaculates each, subsequently divided into four experimental groups: G1 (no CLC), G2 (1 mg CLC), G3 (1.5 mg CLC) and G4 (2 mg CLC). In Experiment I, the semen was cryopreserved according to Papa et al. (Animal Reproduction Science. 2008; 107:293-301), and sperm kinetics, mitochondrial potential and membrane destabilization were evaluated after thawing at 37°C. In Experiment II, the same parameters were evaluated after addition of FertTalp after thawing (20% v/v), as well as the evaluation of intracellular hydrogen peroxide production and a thermo-resistance test (TTR) to verify semen longevity for 120 minutes. Statistical analysis was performed using the F test, Tukey, Friedman and Conover, significance 5%. The results of Experiment I indicated that group G3 (1.5 mg CLC) showed higher total motility (TM) and progressive motility (PM) than group G1 (without CLC) (P&lt;0.05), suggesting that this concentration of cholesterol is efficient in maintaining the kinetic quality of sperm after thawing. There were no significant differences between the groups regarding mitochondrial potential and plasma membrane destabilization. In Experiment II, addition of FertTalp post-thawing resulted in a significant increase in sperm velocity parameters (VCL, VSL, VAP) after 80 minutes of thawing, when compared to groups without FertTalp. The addition of the stimulant also favoured plasma membrane integrity and reduced intracellular hydrogen peroxide production, indicating that FertTalp was beneficial in preserving cell functionality during the thawing process. Data from the TTR showed that, over time (0 to 120 minutes), groups with FertTalp addition maintained higher motility and superior sperm velocity parameters when compared to other groups. These results suggest that the addition of CLC at a concentration of 1.5 mg before cryopreservation can improve sperm quality after thawing, especially concerning the motility. Furthermore, the addition of FertTalp as a post-thaw motility stimulant provides additional benefits, especially in preserving velocity and cellular integrity over time. In conclusion, the addition of CLC is beneficial for improving sperm motility after cryopreservation, and the use of FertTalp provides additional advantages in maintaining sperm quality during thawing and subsequent incubation, promoting greater sperm longevity and viability.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105333"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386707","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The relationship between post-thaw sperm quality and blastocyst production following Intracytoplasmic Sperm Injection (ICSI) of in vitro-matured equine oocytes","authors":"C. Hernandez-Aviles ,&nbsp;C.C. Love ,&nbsp;L. Ramirez-Agamez","doi":"10.1016/j.jevs.2024.105293","DOIUrl":"10.1016/j.jevs.2024.105293","url":null,"abstract":"<div><div>In vitro production of equine embryos by Intracytoplasmic Sperm Injection (ICSI) is commonly utilized to maximize the availability of sperm from stallions with limited semen supply. The relationship between standard measures of sperm quality in frozen/thawed semen (i.e., sperm motility, normal morphology, DNA quality) and embryo production after ICSI of in vitro-matured equine oocytes has not been thoroughly studied. In this study, frozen/thawed semen from 44 stallions utilized in a commercial ICSI program was analyzed for post-thaw total and progressive motility (determined both subjectively and by Computer-Assisted Sperm Analysis [CASA]), morphology features (Differential Interference Contrast [DIC] microscopy), and DNA damage (Sperm Chromatin Structure Assay – SCSA) before and after sperm selection by swim-up. Sperm selected by swim-up were used for Piezo-driven ICSI on 485 in vitro-matured equine oocytes obtained by transvaginal oocyte aspiration (TVA) from 59 mares (85 cycles). The relationship between sperm quality characteristics (before and after swim-up), cleavage (&gt;8 blastomeres at day five [5] post-ICSI), and blastocyst rates (day 7 to 10 post-ICSI) was studied using a stepwise logistic regression model (JMP Pro 17.0; SAS Institute, Cary, NC). Statistical significance was set at P&lt;0.05. Data are presented as mean ± SD. Descriptive parameters of ICSI efficiency in this commercial program included: in vitro oocyte maturation rate: 64 ± 18%; cleavage rate at day 5: 62 ± 22%; blastocyst rate per injected oocyte: 29 ± 25%; blastocyst rate per cleaved oocyte: 46 ± 12%; number of blastocysts produced: 139; blastocyst per TVA/ICSI session: 1.64. No single sperm quality parameter was associated with cleavage rate. Post-thaw sperm quality parameters associated with a positive ICSI outcome (i.e., blastocyst production after TVA/ICSI cycle) included mean total motility before swim-up, determined subjectively (33 ± 14%; odds ratio [OR]: 1.20, 95% confidence interval [CI]: 1.02 – 1.42) or by CASA (26 ± 13%; OR: 1.08, CI: 1.01 – 1.28), and morphologically normal sperm (57 ± 12%; OR: 1.31, CI: 1.10 – 1.57). In contrast, mean ± SD proximal droplets after swim-up (3 ± 2 %; OR: 0.63, CI: 0.41 – 0.94), coiled tails before swim-up (4 ± 1%; OR: 0.25, CI: 0.08 – 0.63), and COMPαt before swim-up (16 ± 8%; OR: 0.74, CI: 0.57 – 0.96) were associated with a negative ICSI outcome (i.e., no blastocyst production after a TVA/ICSI cycle). The current study provides clinically useful data regarding post-thaw sperm quality measures that can be assessed in any laboratory setting to determine the fertility potential of frozen/thawed stallion sperm for ICSI.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105293"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386754","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}