Journal of Equine Veterinary Science最新文献

{"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":"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":"Characterization of sperm traits of donkey semen cryopreserved with different permeable cryoprotectants","authors":"C. Zanardi ,&nbsp;A. Carvalho ,&nbsp;C. Freitas-Dell'Aqua ,&nbsp;M.A. Alvarenga ,&nbsp;F.O. Papa ,&nbsp;G. Monteiro ,&nbsp;L. Segabinazzi","doi":"10.1016/j.jevs.2024.105336","DOIUrl":"10.1016/j.jevs.2024.105336","url":null,"abstract":"&lt;div&gt;&lt;div&gt;Recent research has highlighted the potential advantages of dimethylformamide (DMF) over methylformamide (MF) as cryoprotectants for donkey semen (Bruno.et.al. JEVS 2024;136:105069). Higher concentrations of cryoprotectants could negatively affect mare fertility, underscoring the need for optimized protocols to balance cryoprotection with reproductive outcomes. The objective of this study was to assess (Experiment 1) the effects of different permeable cryoprotectants (glycerol [GLY], etilenoglycol [ETI], MF, and DMF) and (Experiment 2) to investigate the impact of reduced concentrations of cryoprotectants (5%, 3%, and 2%) on sperm traits of cryopreserved donkey semen. Semen was extended to 100 million sperm/mL with a sodium caseinate medium (BotuGold&lt;sup&gt;Ⓡ&lt;/sup&gt;), centrifuged (600xg/10min) and resuspended (100 million sperm/mL) in one of the tested extenders for cryopreservation (Oliveira.et.al., Theriogenology,2016;85:12-67-73). In experiment 1, four ejaculates of five Pêga donkeys (n=20) were cryopreserved using the BotuCrio&lt;sup&gt;Ⓡ&lt;/sup&gt;-based medium (10% of egg yolk) added with 5% of one of the following cryoprotectants: GLY, ETI, MF, or DMF; and BotuCrio&lt;sup&gt;Ⓡ&lt;/sup&gt; (BC, 1% GLY and 4% MF) was used as a Control. In experiment 2, five ejaculates of six Pêga donkeys (n=30) were cryopreserved using BotuCrio&lt;sup&gt;Ⓡ&lt;/sup&gt;-based medium containing 5%, 3%, or 2% of the BC cryoprotectant combination (1:4; GLY:MF). Frozen-thawed semen (37°C/30s) was assessed by CASA for total motility (TM), progressive motility (PM), rapid sperm percentage (RAP), and by fluorescence microscopy for plasma membrane integrity (PMI). Data were assessed using mixed model/Bonferroni for parametric data, and Kruskal-Wallis/Dunns for non-parametric data with P&lt;0.05. In experiment 1, BC and DMF had the highest TM (P&lt;0.05) whereas ETI presented the lowest TM but similar to GLY. MF had intermediate (P&gt;0.05) values of TM only similar to GLY (BC: 59.75±12.49; DMF:60.55±12.75, MF: 50.10±14.47; GLY:45.75±16.11; EG:39.40±17.24). PM was higher in BC compared to GLY and ETI (P&lt;0.05), whereas MF and DMF had intermediate PM results similar to all groups (BC: 39.70±13.12; MF: 33.85±11.88; DMF: 33.30±8.47; GLY: 27.80±12.78; ETI: 29.65±20.25). DMF yielded the best results for RAP, which was similar to BC, whereas ETI and GLY had the lowest RAP values (P&lt;0.05). MF had intermediate RAP parameters compared to all groups (DMF: 48.25±14.40; BC:45.50±16.14; MF: 38.50±15.23; GLY: 31.35±15.67; ETI: 29.55±16.51). DMF and BC better (P&lt;0.05) preserved sperm PMI. MF and GLY had intermediate PMI compared to all groups (DMF: 44.35±9.68; BC: 41.25±10.44; MF: 35.05±10.81; GLY: 40.10±10.38; ETI: 33.55±12.67). In experiment 2, TM was similar across groups (5%: 57.70±4.82; 3%: 53.60±4.82; 2%: 49.50±4.82). However, the 5% yielded higher PM and RAP compared to 2% (P&lt;0.05; PM: 5%: 37.00±6.11; 2%: 29.90±6.11; RAP: 5%: 42.40±7.15; 2%: 34.30±7.15), but similar to 3% (PM, 34.00±6.11; RAP, ","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105336"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386621","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":"Cadmium, lead, and arsenic in stallion semen","authors":"M. Akar ,&nbsp;T. Oztas ,&nbsp;O. Peltoniemi ,&nbsp;S. Goericke-Pesch ,&nbsp;S. Björkman ,&nbsp;M. Kareskoski","doi":"10.1016/j.jevs.2024.105260","DOIUrl":"10.1016/j.jevs.2024.105260","url":null,"abstract":"<div><div>Heavy metals such as cadmium, lead, and arsenic are recognized for their toxic effects on reproductive health across species. These elements can impair spermatogenesis, reduce sperm motility, and cause DNA damage, compromising fertility. Exposure to heavy metals poses a concern for breeding programs, potentially leading to reduced reproductive success and long-term genetic impacts on offspring health. The horse can also be a potential bioindicator of exposure to heavy metals that might not be detected in human blood samples. The aim of this study was to evaluate and compare the concentrations of cadmium, lead, and arsenic in the serum, hair, and semen of stallions. Serum, hair, and semen samples were collected from 17 fertile breeding stallions (mean age 17.5 (13-25) years, during the 2021 breeding season on two commercial stud farms in Western Finland. The stallions were housed in box stalls on peat bedding, with daily access to pasture. Feeding regimens varied between individuals but consisted of locally produced hay or haylage and commercial concentrates. The study was approved by the National Animal Experimentation Board of Finland (ESAVI/22394/2022). One ejaculate per stallion was collected with an artificial vagina, and a 10 mL aliquot of raw semen was frozen at −80°C for further analysis. Serum and hair samples were collected in heavy metal-free serum tubes and stored at −80°C until further analyses. The samples were digested using a microwave-digestion system and analyzed following the guidelines of SFS-EN ISO 17294-2 (2016) on inductively coupled plasma mass spectrometry (ICP-MS) for determining selected elements. Overall, the concentrations of heavy metals were considered low. The concentrations varied between sample types, with hair samples containing the highest levels of As and Pb. The concentrations (mean ± SD) of cadmium in serum, hair, and semen were 0.46 ± 0.17 µg/L, 6.85 ± 4.80 µg/kg, and 0.15 ± 0.08 µg/L, respectively. The concentrations (mean ± SD) of lead in serum, hair, and semen were 2.77 ± 0.93 µg/L, 160.4 ± 114.3 µg/kg, and 0.44 ± 0.39 µg/L. The concentrations (mean ± SD) of arsenic in serum, hair, and semen were 0.95 ± 0.68 µg/L, 131.2 ± 90.2 µg/kg, and 0.57 ± 0.36 µg/L. Seven stallions had serum cadmium values exceeding the exposure limit value for humans (&gt; 0.5 ug/l), but all samples were well below tissue concentrations of clinical toxicity. No statistically significant correlations existed between serum and semen concentrations of arsenic, cadmium, and lead. In conclusion, the risk of harmful effects of heavy metals on male fertility seems low but may vary between geo-industrial areas.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105260"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386957","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":"In vitro assessment of horse-isolated strains of Lactobacillus acidophilus and Ligilactobacillus equi species for fecal microbiota modulation in horses","authors":"A.M. Nogacka ,&nbsp;A. García ,&nbsp;C. G․ de los Reyes-Gavilán ,&nbsp;S. Arboleya ,&nbsp;M. Gueimonde","doi":"10.1016/j.jevs.2024.105341","DOIUrl":"10.1016/j.jevs.2024.105341","url":null,"abstract":"<div><div>Horses are hindgut fermenters that harbor a complex intestinal microbiota (IM) which provides key enzymes aiding in the breakdown of complex carbohydrates present in their herbivorous diet. Therefore, these animals are deeply dependent on their IM for digestion and nutrition. Consequently, IM imbalances may result in alteration of fermentation patterns with impact on the animal health and the risk of disease. In this context, strategies for assisting the maintenance of a healthy IM in horses are of interest. However, there is limited research concerning the use of probiotics to improve hindgut fermentation and diet digestibility, with very few studies focusing on the use of lactobacilli strains from equine origin. Herein, we conducted independent fecal batch fermentations, using feces from “Asturcón” horses as inocula, added individually with four different lactobacilli strains (two strains of <em>Lactobacillus acidophilus</em> and two of <em>Ligilactobacillus equi</em>) isolated from this same horse breed. The impact on the gut microbiota composition was assessed by 16S rRNA gene profiling and the metabolic activity (production of short-chain fatty acids) by gas chromatography. The functionality of the lactobacilli strain was determined by monitoring in real-time gas production and determining changes in pH along incubation. <em>L. acidophilus</em> IPLA20127, promoted an increase in IM diversity and in the relative abundance of <em>Lactobacillus</em> genus, as well as higher butyric and valeric acid levels. This strain shows potential as probiotic supplement, without triggering acidification, nor promoting an increase of gas production or abrupt IM changes in our experimental model.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105341"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142915189","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":"Can a simple and cheap semen assessment help with fertility?","authors":"G.B.A.G. Amorim,&nbsp;O.M. Oliveira,&nbsp;F.O. Papa,&nbsp;J.A. Dell'Aqua Jr,&nbsp;M.A. Alvarenga,&nbsp;C.P. Freitas-Dell'Aqua","doi":"10.1016/j.jevs.2024.105262","DOIUrl":"10.1016/j.jevs.2024.105262","url":null,"abstract":"<div><div>The objective of this study was to evaluate, using light microscopy and simple tests, the following parameters in stallion semen: total and progressive motility, plasma membrane integrity, and sperm morphology, and to determine which of these factors may interfere with fertility. The study utilized a diluent for sperm evaluation based on Eosin and Nigrosin (Botuvital-Botupharma, Botucatu, Brazil), which allows the analysis of sperm pathologies and visualization of plasma membrane integrity under light microscopy. A total of 24 batches of frozen semen from 24 Arabian and Straight Egyptian Arabian stallions were used. Seen was frozen at different locations in Europe and the Middle East, and used for AI in Doha, Qatar. The straws contained the same concentration pattern of 100 million sperm cells. Each batch was thawed at 37°C for 1min, total and progressive motility were evaluated using light microscopy at 40X magnification, Botuvital was used to assess sperm morphology and membrane integrity. A total of 200 cells per sample was evaluated (at 100Xmagnification), and those with a pink head were classified as having a damaged plasmatic membrane. For sperm morphology, using the same light microscope, 200 cells per sample were categorized into major, minor, total, and non-compensatory defects. For AI, mares were examined once per day by transrectal ultrasonography (Mindray DP50 ultrasound with transrectal probe - frequency 5mHz). When a follicle ≥35mm was detected along with uterine edema, a single dose (1ml) of histrelin acetate (250µg, intramuscular, Strelin, Botupharma, Brazil) was administered. Deep-horn uterine insemination was performed 36-40 hours after ovulation induction using an equine universal flexible pipette. A single AI was performed on each mare on top of ovulation, and stallion groups were separated based on whether pregnancy was confirmed or not at 14 days by ultrasound. Parameters were assessed for normality using the Kolmogorov-Smirnov test, variables that showed normal distribution were compared using the unpaired T-test and non-parametric were compared using the Mann-Whitney test, with P&lt;0.05 considered significant. Major sperm pathologies (P=0.0252), total sperm pathologies (P=0.0189), and non-compensatory defects (P=0.0099) - pathologies that cannot be compensated by increasing sperm concentration (Saacke.Theriogenology.2008) - showed statistically significant differences, while total motility, progressive motility, and membrane integrity did not show significant differences between animals that had confirmed pregnancies and those that did not. In conclusion, morphologically abnormal sperm are a critical component in the analysis of semen quality, and this highlights the efficiency of a quick and inexpensive assessment of sperm morphology, which can be conducted in the field.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105262"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386694","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":"Sedation with xylazine and detomidine for andrological examination in horses and reversal with yohimbine: is there an impact on ejaculate quality?","authors":"C.M. Trinque,&nbsp;L.E.F. Canuto,&nbsp;T.M.S. Cavalero,&nbsp;M.S. Frasson,&nbsp;G.S. Camargo,&nbsp;L. Sitó da Silva,&nbsp;M.A. Alvarenga,&nbsp;C.P. Freitas-Dell'Aqua,&nbsp;J.A. Dell'Aqua Jr,&nbsp;F.O. Papa","doi":"10.1016/j.jevs.2024.105327","DOIUrl":"10.1016/j.jevs.2024.105327","url":null,"abstract":"&lt;div&gt;&lt;div&gt;For transrectal palpation and ultrasonography to assess the internal glands of a stallion, veterinarians often use physical or chemical restraint to facilitate the procedure, with the majority using sedation (Pearson et al., Equine Veterinary Education, 2020; 33: 522-530). The objectives of the study were to evaluate the use of a combination of xylazine and detomidine sedation (SEDACOMBO&lt;sup&gt;Ⓡ&lt;/sup&gt;: 0.4 mg/kg of xylazine and 0.008 mg/kg of detomidine, Botupharma Ltda., Botucatu, SP, Brazil) to perform an internal reproductive system examination, as well as the reversal by yohimbine (RESET&lt;sup&gt;Ⓡ&lt;/sup&gt;: 0.1 mg/kg, Botupharma Ltda., Botucatu, SP, Brazil) on the sperm parameters of stallions. Five stallions, aged 4 to 20 years, were assigned to 3 different treatments in a crossover design: CONTROL (saline solution/saline solution), GROUP 1 (SEDACOMBO&lt;sup&gt;Ⓡ&lt;/sup&gt;/RESET&lt;sup&gt;Ⓡ&lt;/sup&gt;), and GROUP 2 (saline solution/RESET&lt;sup&gt;Ⓡ&lt;/sup&gt;). After stimulation by an estrous mare, an intravenous injection of Sedacombo&lt;sup&gt;Ⓡ&lt;/sup&gt; or saline solution was given. After 30 minutes, RESET&lt;sup&gt;Ⓡ&lt;/sup&gt; or saline solution was administered, and after an additional 15 minutes, semen collection was performed. Immediately after collection, the ejaculate was analyzed for volume (with and without gel), total number of sperm, sperm concentration, sperm kinetics (Computer Assisted Sperm Analyses - CASA), membrane stability, mitochondrial potential, and superoxide anion generation via flow cytometry. Statistical analysis was performed using the Kolmogorov-Smirnov, ANOVA, and Tukey tests, with P&lt;0.05 considered significant. GROUP 2 showed a higher seminal volume, with gel (103.7±12.2 ml) and without gel (87.4±8.4 ml), compared to the CONTROL group (62.2±9.9 and 51.8±8.3 ml; P&lt;0.05) and without gel compared to GROUP 1 (P&lt;0.05) (53.6±8.0 ml). Sperm concentration was lower in GROUP 2 (P〈 0.05) (97.8±11.1 × 10&lt;sup&gt;6&lt;/sup&gt;/ml) compared to CONTROL (185.3±27.2 × 10&lt;sup&gt;6&lt;/sup&gt;/ml), but similar to GROUP 1 (167.8±33.7 × 10&lt;sup&gt;6&lt;/sup&gt;/ml). The total sperm number remained the same across all treatments. GROUP 1 demonstrated greater membrane stability (84±1.5%) compared to GROUP 2 (72.0±4.4%). The CONTROL group (83.2±2.5%) and GROUP 1 (84.4±1.1%) had similar numbers of cells with high mitochondrial potential, both more than GROUP 2 (73.6±4.1%). Other parameters (kinetics and superoxide anion generation) did not differ between groups. The increase in ejaculate volume in GROUP 2 can be explained by the presence of yohimbine, which blocks alpha-2 adrenergic receptors in the central nervous system, stimulating the ejaculation process (Yonezawa et al., Biomedical Research, 2005; 26: 201-206). The increase in seminal plasma in GROUP 2 explains the decrease in sperm membrane quality (Monteiro et al., Veterinária e Zootecnia, 2011; 18(2): 255-263). In conclusion, the isolated application of RESET&lt;sup&gt;Ⓡ&lt;/sup&gt; altered sperm membrane integrity, but when applied 30 minutes after sedati","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105327"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386703","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}

{"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}