Journal of Equine Veterinary Science最新文献

{"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}

{"title":"BeyondⓇ extender in donkey semen: evaluation at 5 °C and 17 °C over 7 days","authors":"M. Caldevilla,&nbsp;D. Neild,&nbsp;A. Ferrante,&nbsp;I. Cáceres Monie,&nbsp;C. Arraztoa,&nbsp;J. Plaza,&nbsp;J. Otero,&nbsp;M. Gambarotta,&nbsp;M. Miragaya","doi":"10.1016/j.jevs.2024.105270","DOIUrl":"10.1016/j.jevs.2024.105270","url":null,"abstract":"<div><div>The objective of this study was to evaluate the ability of the equine commercial extender Beyond^Ⓡ, both alone and with the addition of fresh egg-yolk (EY), to maintain donkey sperm motility, viability and acrosomes at 5 °C and 17 °C over 7 days or until progressive motility (PM) was lower than 20%. Fifteen ejaculates from 5 fertile jacks were diluted in EquiPlusTM, incubated 30 minutes at room temperature, centrifuged and resuspended in: Beyond^Ⓡ(B), B with 2% EY (BY) and skim milk-based extender with 2% EY (KY). Sperm kinematic parameters (CASA; AndroVision^Ⓡ) and viability and acrosome integrity (FITC-PNA/PI using flow cytometry; BD FACSCanto II^Ⓡ) were evaluated daily. Either ANOVA or Kruskal Wallis was used to analyze the data and P&lt;0.05 was considered significant. In the samples at 5 °C a significant decrease in % PM was observed between day 7 (B: 34.18±8.0; BY: 36.14 ±17.9; KY: 30.08±0) compared to 24h (B: 50.04±20.8; BY: 59.19±20.4; KY: 63.29±17.1). At 72h significantly higher percentages of total motility (TM) were observed for B and BY compared to KY. PM in B was not significantly different to BY and KY at each evaluation time. However, the number of samples with PM &gt; 20% at day 7 were greater in the BY samples (9/15) compared to B (5/15) and KY (1/15). Additionally, a significant decrease in live acrosome intact (LAI) sperm was observed on day 7 (B: 19.55±15.63; BY: 9.99±10.03; KY: 36.79±0) compared to 24h (B: 43.24±19.18; BY: 55.56±16.96; KY: 61.89±12.26). No significant differences were observed in % LAI for B compared to BY and KY at each evaluation time. At 17 °C, a significant decrease in percentage PM was observed on day 7 (B: 33.5±11.55; BY: 36.69±8.6) compared to 24h (B: 60.47±15.1; BY: 65.57±13.1). TM and PM in B were not significantly different to BY at each evaluation time. The number of samples with PM &gt; 20% at day 7 was greater in the BY samples (9/15) compared to B (3/15). Additionally, % LAI was significantly lower on day 7 (B: 26.67±28.82; BY: 14.49±14.65) compared to 24h (B: 45.40±28.68; BY: 48.39±28.83). No significant differences were observed in % LAI for B compared to BY at each evaluation time. No significant differences were observed in any of the sperm parameters between both temperatures (5 and 17 °C) for any of the extenders assayed. To conclude, although Beyond^Ⓡ can be used to preserve donkey semen for 7 days, both at 5 and 17 °C, it is noteworthy that many more samples lasted 7 days with PM &gt; 20% when extended in Beyond^Ⓡ with 2% egg-yolk.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105270"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386616","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":"Comparison of techniques for sperm evaluation in Purebred Spanish Horses","authors":"N. Latorre ,&nbsp;A. Sánchez-Rodríguez ,&nbsp;C. Gómez-Cuétara ,&nbsp;E.R.S. Roldan","doi":"10.1016/j.jevs.2024.105296","DOIUrl":"10.1016/j.jevs.2024.105296","url":null,"abstract":"<div><div>The present study was designed to compare methods of sperm assessment in Purebred Spanish Horses, to evaluate different staining methods and to facilitate routine handling and work both in research laboratories and in the field. The parameters evaluated were acrosome integrity, capacitation, and sperm protamination. Semen was collected from February to May 2024 from 11 PRE stallions (three replicates of each). Immediately after collection, samples were diluted in INRA96 to a final concentration of 100 × 10⁶ sperm/mL and maintained at room temperature. For the examination of acrosome integrity, smears stained with Eosin-Nigrosin-Giemsa (ENG) were compared with smears stained with peanut agglutinin (PNA). Capacitation was evaluated by analyzing smears stained with PNA compared with samples fixed in 2% glutaraldehyde in cacodylate buffer and stained with Hoechst 33258 and chlortetracycline (CTC). Finally, for the analysis of protamination, smears stained with chromomycin A3 (CMA3), methylene blue (Diff-Quik), aniline blue, or toluidine blue were compared. Significant differences (p&lt;0.05) between techniques were found using a paired t-test, when comparing two groups, or one-way-ANOVA, when comparing four groups; correlation tests were also performed (GraphPad Prism v9). The percentage of sperm with intact acrosomes identified with ENG was higher than with PNA, whereas sperm with damaged acrosomes with ENG was lower than with PNA. Conversely, the percentage of sperm with lost acrosomes was similar between the two techniques. Microscopic evaluation of sperm stained with ENG was more complex and time-consuming compared with PNA staining. The percentage of capacitated sperm analyzed with PNA and CTC, showed no significant differences between the two methods. For these two parameters, the correlations between the methods studied were not significant. Finally, chromatin compaction alterations showed significant differences between aniline blue (1.3±0.6%) and both Diff-Quik (4.0±0.7%) and toluidine blue (3.5±0.4%), whereas CMA3 (2.4±0.5%) were similar when compared with the other methods. The only significant correlation (0=0.019) appeared between CMA3 and aniline blue (Pearson r=0.69). It is noteworthy that these staining techniques evaluate different aspects of the chromatin, which may be the reason of the observed differences. Therefore, a reference technique, such as SCSA, would be a good option for the comparison of these 4 methods. Future studies should focus on the identification of a method of reference for each parameter to perform regression studies that allow us to model differences and generate an algorithm so the different staining techniques may be used in different laboratories or in the field and be comparable.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105296"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386760","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":"Prediction of breeding soundness in pre-, peri- and postpubertal colts","authors":"Martin Köhne ,&nbsp;Astrid von Rintelen-Feldmann ,&nbsp;Martha Papkalla ,&nbsp;Anna Tönissen ,&nbsp;Gunilla Martinsson ,&nbsp;Marion Schmicke ,&nbsp;Harald Sieme","doi":"10.1016/j.jevs.2024.105295","DOIUrl":"10.1016/j.jevs.2024.105295","url":null,"abstract":"<div><div>Prediction of future fertility is important for stallion breeders as rearing of colts for breeding purposes is expensive. Thus, the study examined the testicular development and hormonal profiles in a group of pre-, peri- and postpubertal Warmblood colts (n=46) and evaluated the fertility of selected stallions (n=12) at 2.5 years of age, aiming to find parameters for predicting the breeding soundness at the youngest possible age. Exams were performed at 177±23, 604±23 and 894±24 days of age and included body condition scoring and weighing, examination of the testes (palpation, ultrasound, volume determination) and blood sampling (before and after stimulation with hCG (5000 I.U., i.v.). Hormone analysis included determination of AMH (basal) and testosterone (at 0, 2, 24 and 72 h after stimulation). Stallions with abnormal (n=6) and normal (n=6) were subjected to spermatological examination. During the examination period, palpatory findings were inconsistent in individual colts and testicular volume increased gradually with age. Body mass was 455±38 kg for peripubertal (BCS: 5.17±1.02) and 526±40 kg for postpubertal stallions (BCS: 5.26±0.94). Stallions (n=12) were grouped according to their total testicular volume (small vs. normal) and palpatory findings (abnormal vs. normal) at 2.5 years of age (937±21 days). No significant differences were observed for the total number of morphologically normal, progressively motile spermatozoa for different stallion groups, but more progressively motile spermatozoa (PMS) were observed after storage for 24 h in a centrifuged extended semen sample (p&lt;0.05). Flowcytometric analysis revealed a lower percentage of membrane intact spermatozoa after incubation with Ca-ionophore for 120 min in stallions with small testes, while the percentage of acrosome reacted spermatozoa was higher in this group. A positive correlation of BCS and PMS was found (extended sample: r=0.765, centrifuged sample: 0.699, p&lt;0.05). Analysis of AMH plasma concentrations showed no differences between abnormal or small testes irrespective of age group. AMH concentration was significantly higher in peripubertal colts as compared to younger and older animals. hCG stimulation affected testosterone in all age groups and resulted in the strongest increase in postpubertal stallions 72 h after administration. Based on the evaluated parameters in pre- and peripubertal colts, no prediction of sperm quality at 2.5 years of age was possible. Moreover, inconsistency in palpatory findings (except for cryptorchism) and testicular size for individual colts at 0.5 and 1.5 years of age renders early prediction of breeding soundness in stallions impossible. The results demonstrate, however, an influence of the BCS on sperm quality in postpubertal colts.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105295"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386756","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":"Testicular hormones production and the gene expression of their receptors in the testicles of stallions subjected to scrotal heat stress and treated with pentoxifylline","authors":"Y.F.R. Sancler-Silva ,&nbsp;E.R. Silva-Junior ,&nbsp;A. Esteller-Vico ,&nbsp;Y.L. Boakari ,&nbsp;H. El-Sheikh Ali ,&nbsp;F.O. Papa ,&nbsp;B.A. Ball","doi":"10.1016/j.jevs.2024.105322","DOIUrl":"10.1016/j.jevs.2024.105322","url":null,"abstract":"<div><div>Depending on the duration and intensity of scrotal heat stress, the impact on different cells of the testicular parenchyma varies in severity, initially compromising spermatogenesis and, in more severe cases, impairing hormonal production. Pentoxifylline has been shown to reduce histological damage from testicular degeneration in horses, but its impact on testicular hormonal production after heat stress is still unknown. This study evaluated the effects of heat stress and pentoxifylline treatment on serum concentrations of testosterone, estrone sulfate, and anti-Müllerian hormone (AMH), as well as on the gene expression of their receptors in testicular biopsies of stallions subjected to scrotal heat stress. A total of 14 stallions were divided into three groups: Control (CRL, n=4), Testicular Degeneration (DEG, n=5), and Testicular Degeneration Treated with Pentoxifylline (DEG+PTX, n=5). Testicular degeneration was induced by scrotal insulation using a thermal bag filled with air at 50°C for 1 hour, twice a day (early morning and late afternoon), over two consecutive days (D-1 and D0). From the following day (D1), oral pentoxifylline (17 mg/kg) was administered every 12 hours for 30 days. Blood samples were collected by external jugular venipuncture once a week for eight consecutive weeks. Serum concentrations of testosterone, AMH, and estrone sulfate were determined by ELISA. On days 30 (D30) and 60 (D60), testicular biopsies were taken and snap-frozen for later evaluation by RT-qPCR. The genes evaluated were AR, ESR1, ESR2, and AMH. Normally distributed data were analyzed using a mixed-model ANOVA, and non-normally distributed data were analyzed using the Wilcoxon test, using JMP 12.0 software (SAS Institute, Cary, NC, US). No significant differences were observed between the experimental groups in serum concentrations of testosterone, estrone sulfate, and AMH over the 60-day evaluation period, and all values were consistent with reference values for healthy animals. However, after 60 days of thermal injury, there was a significant increase in AMH gene expression in the pentoxifylline-treated group compared to the control group (P&lt;0.05). No other differences were observed between groups for the other genes evaluated. In conclusion, the heat stress induced in this study did not impact gonadal hormone production or the gene expression of hormonal receptors in the testes. Furthermore, pentoxifylline appears to exert an effect on the upregulation of the AMH gene after 60 days of treatment.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105322"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386622","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":"Changes in equine testicular and epididymal homeostasis in response to scrotal heating","authors":"M. Ferrer ,&nbsp;M. Barletta ,&nbsp;J. Williams ,&nbsp;K. Moran ,&nbsp;G. Bilbao ,&nbsp;J. Bartolome","doi":"10.1016/j.jevs.2024.105284","DOIUrl":"10.1016/j.jevs.2024.105284","url":null,"abstract":"<div><div>Heat stress is a common cause of testicular and epididymal dysfunction. While the effect of scrotal heating on ejaculated sperm and testicular histologic changes is known, the testicular and epididymal molecular events and changes in gene expression that cause them are largely unknown. Here, we hypothesized that early equine testicular and epididymal responses to heat stress are associated with oxidative stress and immune dysregulation. Normospermic adult light-breed stallions were divided into two treatments: control (CON, n=5) and scrotal insulation (SI, n=5). The scrotum of SI stallions was covered with an insulation bag for 24 h. The stallions were castrated immediately after the bags were removed. CON stallions did not undergo scrotal insulation. Tissue biopsies were collected from the testes, head, body, and tail of the epididymis for transcriptome analysis. The RNA sequencing libraries were generated using NEBNext^ⓇUltra™RNA Library Prep Kit (Illumina^Ⓡ). Differential gene expression was compared between each SI and CON tissue using the DESeq2 R package and the ClusterProfiler sofware. Differentially expressed genes (DEGs) and pathways were identified (P&lt;0.05, log2 fold change ≥1) in the PANTHER Classification System. The number of DEGs in the testes, head, body, and tail of the epididymis of SI stallions was 507, 1198, 3420, and 1245, respectively. The main downregulated testicular pathways were associated with spermatogenesis. The SI upregulated testicular DEGs corresponded to 331 pathways (268 biological processes, 80.9%, 42 molecular functions, 12.7%, 21 cellular components, 6.3%). Upregulated DEGs were associated with oxidant-antioxidant balance (e.g. peroxidase activity GO:0004601), increased cell respiration, increased use of lipids and amino acids, purine metabolism, activation of the innate and adaptive immune response (e.g. activation of immune response GO:0002253; positive regulation of innate immune response GO:0045089; T cell activation GO:0050863; B cell proliferation GO:0030888; positive regulation of mast cell activation GO:0033005; e.g. FGR, LRRC32, CD226, TLR6, TRIM15, IL33, CD4), and deposition of amyloid. The epididymal head had dysregulation of pathways associated with spermatogenesis, energy metabolism, and antigen presentation and processing, with DEGs associated with macrophage and T cell signaling, regulatory T cell differentiation, and cytotoxic T cells. DEGs in the epididymal body were associated with microtubule formation, sperm flagellum components, and cell motility. DEGs in the epididymal tail were associated with energy metabolism and cell respiration. The findings support dysregulation of the local oxidant-antioxidant system, energy metabolic pathways, and immune system homeostasis in SI stallions and lay the foundation for understanding heat-induced reproductive dysfunction.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105284"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386844","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":"Readers Information","authors":"","doi":"10.1016/S0737-0806(25)00031-0","DOIUrl":"10.1016/S0737-0806(25)00031-0","url":null,"abstract":"","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105373"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386508","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":"Variation in semen lipids during cryopreservation by cooling with milk-based extender in donkeys","authors":"M.M.C. Chaves ,&nbsp;C. Ramires Neto ,&nbsp;Camila P. Freitas-Dell'Aqua ,&nbsp;K.R. Belaz ,&nbsp;M.N. Eberlin ,&nbsp;F.O. Papa ,&nbsp;M.J. Sudano ,&nbsp;I.F. Canisso ,&nbsp;M.A. Alvarenga ,&nbsp;J.A. Dell'Aqua Jr","doi":"10.1016/j.jevs.2024.105275","DOIUrl":"10.1016/j.jevs.2024.105275","url":null,"abstract":"<div><div>Donkey sperm cryopreservation is becoming increasingly necessary for the production of hybrids with horses and seed stock of endangered breeds. Therefore, this study aimed to assess lipid changes during the cryopreservation of donkey sperm. One ejaculate from each of seven donkeys was harvested, evaluated, and extended in a skim milk-based extender at 50 million sperm/mL and cooled for 24h at 5°C. Aliquots of fresh, not extended semen were preserved in liquid nitrogen for lipidomic analyses by MALDI-MS. The analyses were performed with MetaboAnalyst 2.0 and the discriminating analyses of PLS-DA to demonstrate lipid variation occurring during the cryopreservation process. The lipid types that were differentially expressed were analysed by T-test followed by Tukey. The Vulcano plot was used to compare groups, using the following criteria values of P&lt;0.05 and fold-change (FC) ≥1.5. The identification of lipids was in the databases <span><span>www.lipidmaps.org</span><svg><path></path></svg></span>. Analysis of phospholipids between fresh and cooled semen showed 96 total lipids in fresh semen and 98 in cooled semen; 18 lipids are most abundant in the fresh semen, of which 6 are Glycerophospholipids, 2 Glycerolipids, 2 Sphingolipids, 2 are Fatty Acyls and 1 (728.6 m/z) can be Sphingolipids or Glycerolipids, 8 of which are known to be polyunsaturated and 4 saturated and the other 5 were not identified (733.9, 748, 764, 801, 826 m/z); 14 lipids are most abundant in refrigerated semen, 1 Sterol Lipids, 7 Glycerophospholipids, 1 Glycerolipids, 1 Fatty Acyls, 2 Sphingolipids, of these 11 are polyunsaturated and 1 saturated, another 2 (737, 749 m/z) were not identified. After refrigeration, 14 lipids were lost, 1 Sterol Lipids, 5 Glycerophospholipids 1, Sphingolipids and 4 Glycerolipids, 8 unsaturated and 4 saturated and 3 (736, 760, 762 m/z) were not identified, and even after refrigeration 16 lipids were incorporated on semen, 6 Glycerolipids, 1 Glycerophospholipids and 3 Fatty Acyls, of which 9 were unsaturated and 1 saturated, the remaining 6 (737, 755, 759, 798, 811, 859 m/z) lipids were not identified. Comparatively, refrigerated semen showed a greater presence of polyunsaturated lipids compared to fresh semen and the addition of the milk-based extender incorporated more unsaturated lipids into the sperm membrane. The presence of double bonds between carbons in polyunsaturated fatty acid molecules makes them very vulnerable to attack by free radicals and the initiation of the lipoperoxidation cascade. In conclusion, freezing donkey semen results in a change in phospholipids with loss of lipids, incorporation of new ones and change in relative abundance mainly of polyunsaturated lipids that can lead sperm to greater oxidative damage during the cryopreservation process.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105275"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386514","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}