Journal of Equine Veterinary Science最新文献

{"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":"Unraveling the effect of glycerol and amides and different cooling rates on post-thawed sperm parameters of equine frozen semen","authors":"M.S. Frasson ,&nbsp;S.N. Oliveira ,&nbsp;J.A. Dell'Aqua Jr ,&nbsp;L.T. Rodrigues ,&nbsp;T.M.S. Cavaleiro ,&nbsp;C.P. Freitas-Dell'Aqua ,&nbsp;A.M. Crespilho ,&nbsp;P. de Mello Papa ,&nbsp;L. Segabinazzi ,&nbsp;F.O. Papa","doi":"10.1016/j.jevs.2024.105286","DOIUrl":"10.1016/j.jevs.2024.105286","url":null,"abstract":"<div><div>This study assessed the effect of different cooling curves, stabilization periods, and permeating cryoprotectants on equine cryopreserved semen. Forty-eight ejaculates of six stallions were used for the study. After semen collection, the ejaculates were split into four aliquots, extended 1:1 (v:v, semen:extender) with a milk-based extender, and centrifuged at 600 × g for 10 min. After centrifugation, the sperm pellet was resuspended at 200 × 106 sperm/mL in a BotuCrio^Ⓡ-based medium containing 5% of one of the following permeating cryoprotectant: glycerol (GLY), methylformamide (MF), dimethylformamide (DMF) and dimethylacetamide (DMA). Thereafter, semen was packed into 0.5 mL French straws and five straws from each group were subjected to four cooling rates using the Mini-Digitcool ZH 400 system: -1°C/min(C1), -0.25°C/min (C2), -4°C/min with stabilization at 5°C for 15min (C3), and -4°C/min with stabilization at 5°C for 75min (C4). Afterwards, the straws were frozen in nitrogen vapor (-140°C) at curve of ∼ -25°C/min with stabilization for 20min, and then immersed in liquid nitrogen and stored at -196°C. For sperm analysis, frozen semen samples were thawed at 46°C for 20s, incubated at 37°C/10min, and assessed for sperm kinetics by the CASA system (Hamilton Thorne Research – IVOS 12) and for plasma membrane stability (PMS) and high mitochondrial membrane potential (HMMP) by flow cytometry. Sperm parameters between cryoprotectants and cooling curves were assessed by ANOVA and Tukey test, and P was set as 〈 0.05. The C1 cooling curve produced superior sperm parameters independent of the cryoprotectant (TM, 60±12; PM, 21±12; RAP, 37±17; PMS, 50±13; HMMP, 6858±1230) compared to C2 (TM, 56±16; PM, 19±13; RAP, 33±18; PMS, 48±9; HMMP, 4827±1053), C3(TM, 54±15; PM, 17±11; RAP, 33±16; PMS, 41±9; HMMP, 2131±1043) e C4 (TM, 55±15; PM, 17±13; RAP, 33±16; PMS, 39±10; HMMP, 2112±1010). GLY samples had superior sperm kinetics (TM, 58±18; PM, 28±13; RAP, 40±19) and for HMMP (4504±2472), but lower for PMS (39±9) than DMF (TM, 60±10; PM, 17±7; RAP, 38±9; HMMP, 3709±1961; PMS 51±11) and DMA (TM, 51±15; PM, 9±6; RAP, 23±15; HMMP, 3719±2186; PMS, 46±11), but similar to MF (TM, 58±13; PM, 21±13; RAP, 36±17; HMMP, 4015±2377; PMS,41±9). DMA samples had the lowest values of sperm kinetics, whereas DMF presented the highest plasma membrane stability. In conclusion, GLY presented superior sperm kinetics and HMMP to the amides (DMF and DMA), and mainly DMF presented greater protection to the plasma sperm membrane. Regardless of the cryoprotectant added, the quickest freezing curve, -1°C/min, was more effective in preserving sperm quality.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105286"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386846","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":"Effect of heterologous blood plasma (HBP) on the freezing extender of equine sperm","authors":"L. Tobal,&nbsp;L. Arruda,&nbsp;G. Pinto,&nbsp;G. Xavier,&nbsp;M. Guerra,&nbsp;G. Carneiro","doi":"10.1016/j.jevs.2024.105326","DOIUrl":"10.1016/j.jevs.2024.105326","url":null,"abstract":"<div><div>Sperm cryopreservation is essential for the conservation of genetic resources, but freezing can compromise its fertilizing capacity. Optimizing cryopreservation techniques is crucial for developing effective extenders. Platelet-rich plasma (PRP) is an innovative therapeutic approach, showing promising results. However, considering the difficulty of performing absolute platelet counts in the field, this study integrated heterologous blood plasma (HBP) into the cryopreservation extender for equine semen. The objective was to evaluate the efficacy of HBP, added to freezing extender, through analysis of sperm kinetic parameters, plasma membrane integrity (PMI), acrosomal membrane integrity (AMI), and mitochondrial membrane potential (MMP). The experiment was conducted at the Animal Andrology Laboratory of the Federal Rural University of Pernambuco, using five stallions (5 to 9 years old). Blood samples were collected in tubes containing 3.6% sodium citrate and subjected to centrifugation (100G/10 minutes). Supernatant was collected and centrifuged (1600G/20 minutes) and combined into a single tube and homogenized. Ejaculates were collected using an artificial vagina. For freezing, samples were diluted in Botucrio^Ⓡ, supplemented with HBP (0%, 1%, 2%, and 5%), and frozen in an automated system (TK 3000^Ⓡ). Statistical analyses were performed using GraphPad InStat software. No significant differences were observed in total motility parameter among the treated groups. However, the study revealed significant differences in lateral head amplitude (ALH) parameters, with the group treated with 5% HBP showing higher values than control group. Additionally, linearity (LIN) values were greater in the 1.25% and 5.0% groups compared to control group (P ≤ 0.05). Kinetic parameters such as LIN and WOB have been shown to have a high correlation with the progressivity of frozen-thawed sperm (Harris. et al. Reproduction. 2023). Conversely, the wobble index (WOB) demonstrated a significant increase compared to control group. Regarding acrosome membrane integrity (%), it was seen a positive effect of HBP in the groups with concentrations of 2.5% (65,12± 6.01) and 5% (65,64± 3.80) when compared to control group (61,44±7.17). These results can be attributed to the buffering effect of HBP, that prevents osmotic shock, decreasing the risk of crystallization during the cryopreservation process. It can be concluded that adding heterologous blood plasma to the freezing medium demonstrated satisfactory results in sperm kinetics, presenting linear sperm with an intact acrosome.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105326"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386702","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 isoform lactate dehydrogenase C (LDHC) has a major role in improving stallion spermatozoa incubated in pyruvate-based media","authors":"E. da Silva Álvarez,&nbsp;L. Becerro Rey,&nbsp;F.E. Martin Cano,&nbsp;A. Silva Rodríguez,&nbsp;C. Ortega Ferrusola,&nbsp;G. Gaitskell-Phillips,&nbsp;M.C. Gil Anaya,&nbsp;F.J. Peña Vega","doi":"10.1016/j.jevs.2024.105278","DOIUrl":"10.1016/j.jevs.2024.105278","url":null,"abstract":"<div><div>Stallion spermatozoa use different energy sources; while oxidative phosphorylation predominates, glycolysis and beta-oxidation of fatty acids are also important. Moreover, interactions among different pathways are of major importance. Glycolysis depends on the availability of NAD+ electron acceptor, which is reduced to NADH, that in the complex I of the electron transport chain (ETC) donates an electron regenerating NAD+. On the other hand, if mitochondria are damaged, regeneration of NAD+ may be reduced leading to reduced glycolysis further altering sperm metabolism. However, alternative ways to regenerate NAD+ may be present. We hypothesized that aerobic glycolysis is present in the stallion spermatozoa, constituting a backup mechanism to regenerate NAD+. To test this hypothesis, we incubated stallion spermatozoa in different media, including a high glucose media of 67 mM with 1 mM pyruvate and 10 mM glucose with 10 mM pyruvate. The addition of 10 mM pyruvate to the 67mM glucose media improved sperm motility (<em>P&lt;</em>0.001) Aliquots extended in the high glucose media (67 mM glucose) after 3 h of incubation at 37°C experienced a significant drop in motility respect initial values (58.1 ± 1.8% vs 81.2 ± 1.8%; <em>P&lt;</em>0.0001), while aliquots incubated in the 67mM glucose 10 mM pyruvate media, maintained the motility all along the incubation period (77.1± 1.4%), viability and mitochondrial membrane potential were also improved (P&lt;0.001). We studied the metabolic proteome and metabolome using UHPLC/MS/M and identified for the first time three different isoforms of the enzyme lactate dehydrogenase (LDH), LDHA (cytosolic), LDHB (mitochondrial, with higher affinity for pyruvate), and LDHC (cytosol, motile cilium), the latter was the predominant isoform detected. We performed a custom analysis of the stallion sperm proteome using human orthologs in Metascape (<span><span>https://metascape.org/gp/index.html#/main/step1</span><svg><path></path></svg></span>), using the terms “pyruvate” and “lactate” to study its enrichment. Both terms were highly enriched in the stallion spermatozoa; LACTATE <em>p</em>= 1.7 × 1e-13 and PYRUVATE <em>p</em>=4.2 xe-13. Metabolic analysis showed that including 10 mM pyruvate, induced aerobic glycolysis as increased amounts of Lactate and NAD+ were detected. We concluded that activation of aerobic glycolysis in a high glucose media improves sperm survival through the regeneration of NAD+.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105278"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386517","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 three different sized SephadexⓇ beads for filtration of cryopreserved epididymal sperm","authors":"P. Razquin,&nbsp;J. Funes,&nbsp;M,&nbsp;J. Graham","doi":"10.1016/j.jevs.2024.105318","DOIUrl":"10.1016/j.jevs.2024.105318","url":null,"abstract":"&lt;div&gt;&lt;div&gt;Cryopreserving epididymal spermatozoa is often the last opportunity to preserve genetic material for stallions that suffered catastrophic injury, acute or chronic illness, and unexpected death. Epididymal sperm tend to have lower quality and pregnancy rates when compared to cryopreservation of ejaculated frozen semen. Gradient chromatography columns using Sephadex&lt;sup&gt;Ⓡ&lt;/sup&gt; beads have been used to improve the quality of stallion semen; but no data for epidydimal sperm exists. The aim of this study was to compare the efficacy of different Sephadex&lt;sup&gt;Ⓡ&lt;/sup&gt; bead sizes to improve the quality of frozen epididymal sperm. Sperm from adult stallions (n=10) was obtained by retrograde flush post-castration and frozen in a lactose-EDTA extender. The sperm were thawed and evaluated for total and progressive motility, concentration and viability (control). Sephadex&lt;sup&gt;Ⓡ&lt;/sup&gt; columns were prepared (G-25, G-50 and G-75) and sperm from each stallion divided into three equal sized aliquots, which were filtrated through each one of the columns. The filtrate was assessed for volume, sperm concentration, total and progressive motility, morphology and sperm viability. Similar percentages of sperm were recovered from all three sized Sephadex&lt;sup&gt;Ⓡ&lt;/sup&gt; beads (mean 35-37% ±19). The total motility was not significantly different between the control sperm and the Sephadex&lt;sup&gt;Ⓡ&lt;/sup&gt; column groups (mean 35% ±13; 39% ±11; 37% ±12; 36% ±16; for control, G-25, G-50 and G-75). The progressive motility of the control and Sephadex&lt;sup&gt;Ⓡ&lt;/sup&gt; groups were also similar (mean 22%±13; 28%±12; 28%±13; 28%±15; for control, G-25, G-50 and G-75, respectively). Morphological evaluations were similar between control and filtered semen for the percentage of normal sperm (mean 15% ±8; 21% ±9; 22% ±10; 25% ±9), detached heads (mean 2% ±1; 3% ±2; 1% ±1; 2% ±1; p 〉0.05), abnormal heads (mean 28%±13; 25%±16; 25%±17; 22%±16), distal droplets (mean 14%±10; 27%±12; 29%±15; 27%±16), proximal droplets (mean 15% ±6; 11% ±5; 11% ±6; 9% ±6), midpiece defects (mean 1% ±1; 2% ±2; 2% ±2; 3% ±2) and coiled tails (mean 1%±1; 1%±2; 1%±1; 1%±2). The only statistically significant difference was the percentage of sperm with bent tails between control group (mean 22%±17) and Sephadex&lt;sup&gt;Ⓡ&lt;/sup&gt; treatments (mean 8%±7; 7%±5; 8%±; p≤0.05) but not within Sephadex&lt;sup&gt;Ⓡ&lt;/sup&gt; treatments. For viability, a difference between control and Sephadex&lt;sup&gt;Ⓡ&lt;/sup&gt; treatments was observed (mean 52%±12; 66%±8; 57%±9; 57%±10; p≤0.05) and post hoc analysis showed the difference originating from Sephadex&lt;sup&gt;Ⓡ&lt;/sup&gt; G-25 (p≤0.05) with no difference between control and Sephadex&lt;sup&gt;Ⓡ&lt;/sup&gt; G-50 and G-75. Our results show that filtering frozen thawed epidydimal sperm through Sephadex&lt;sup&gt;Ⓡ&lt;/sup&gt; does not improve semen quality although when sperm was filtered through G-25 column there was a reduction in bent tails and an increase in viability. These results differ from other studies where fresh and cooled semen","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105318"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386764","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":"Screening for antimicrobial resistant bacteria in cooled stallion semen and post-insemination uterine lavage fluid","authors":"J. Marttila,&nbsp;E. Tukia,&nbsp;A. Heikinheimo,&nbsp;M. Kareskoski","doi":"10.1016/j.jevs.2024.105303","DOIUrl":"10.1016/j.jevs.2024.105303","url":null,"abstract":"&lt;div&gt;&lt;div&gt;Cooled semen contains antibiotics to control bacterial growth during storage, which may contribute to the development of antimicrobial resistance (AMR) and introduce resistant bacteria into the uterus during artificial insemination (AI). Our aim was to evaluate the presence of AMR in cooled semen and post-insemination uterine lavage fluid. In Part 1, 98 commercial domestic or international cooled semen shipments were cultured after storage ranging from 4h to overnight. Seven samples were cultured both before and after cooled storage with or without antibiotic-containing extender, within 4h and 30 h after semen collection. In Part 2, culture samples were obtained from 10 cooled AI doses that were used for insemination of 10 mares. Uterine lavage fluid samples were collected at 16 to 25 h after insemination. Samples were cultured on sheep blood agars. After enrichment, samples were also cultured on selective media for detection of extended spectrum beta-lactamase (ESBL) producers, carbapenemase-resistant &lt;em&gt;Enterobacteriaceae&lt;/em&gt; (CARBA), methicillin-resistant &lt;em&gt;Staphylococcus aureus&lt;/em&gt; (MRSA), and vancomycin-resistant enterococci (VRE). Bacterial species were identified with MALDI-TOF mass spectrometry. Antibiotic susceptibility of selected pure cultures was determined using the disk diffusion method. In Part 1, 23% of semen samples showed bacterial growth on blood agars. Growth on selective media was observed in 21% (MRSA; none were identified as S. aureus), 17% (ESBL), 14% (CARBA), and 9% (VRE, including &lt;em&gt;Enterococcus faecalis&lt;/em&gt;) of samples. Only 1/7 of the samples stored without extender showed no growth on any medium. In Part 2, most mare and semen samples (9/10) had no bacterial growth. Bacterial growth on selective media was detected in two semen samples: one with growth of clinically non-significant bacterial species on all selective media, and one with non-significant growth on MRSA media. These semen doses were not associated with bacterial growth in corresponding uterine lavage samples. Bacterial growth was observed in two mare samples: one with non-significant growth on MRSA media and one with Enterobacter cloacae -growth on ESBL media. The corresponding semen samples had no bacterial growth. All Enterobacteriaceae-strains isolated from the positive mare sample on ESBL media were resistant to third generation cephalosporins, with three strains producing AmpC-cephalosporinases and one strain producing both ESBL and AmpC-enzymes. Resistant strains, such as ESBL and AmpC-producing &lt;em&gt;Enterobacter cloacae&lt;/em&gt; and VRE may present health risks, including zoonotic spread of AMR. These ESBL and AmpC-enzymes are often plasmid-encoded, facilitating horizontal gene transfer among bacteria, potentially leading to spread of AMR in gram-negative bacteria in horses, complicating infection treatment and posing public health risks due to close human-horse interactions. In conclusion, while bacteria, including resistant strains, m","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105303"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386768","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":"Trends in reproductive technology use in the American Quarter Horse (AQHA)","authors":"P. Loomis,&nbsp;D. Scofield,&nbsp;E. Squires","doi":"10.1016/j.jevs.2024.105300","DOIUrl":"10.1016/j.jevs.2024.105300","url":null,"abstract":"<div><div>The acceptance of reproductive technology by a breed association is dependent on the cost of the procedure, interest in the technology by the breeder as well as the safety and efficacy of the procedure. The AQHA is the largest Breed Registry in the world with 3.1 million registered horses in 2023 and is an example of a registry that has embraced nearly all types of assisted reproduction techniques that have been developed. The AQHA registered 86,762 foals in 2023; 70,526 of these foals were born in the USA and 16,162 foals were born outside the USA. Approximately 8,500 AQHA registered foals were born in Europe, with the remaining international foals born primarily in Canada and Mexico. A significant number of Quarter Horse foals in Australia and South American countries were conceived by frozen semen exported from AQHA registered stallions standing in the US. These countries maintain their own Quarter Horse registries and therefore those foals are not reflected in the AQHA registry data. Data were obtained from the AQHA on the number of registered foals produced each year from embryo transfer (fresh and shipped embryos), frozen embryos, transported cooled and frozen semen. According to the AQHA data, the first foal registered by embryo transfer was in 1981. The number produced remained relatively low until the year 2000, when registrations increased dramatically to over 6000 in 2023. This was due in part to the development of commercial flushing and storage medias that allowed embryos to be collected on the farm and sent to large recipient stations for transfer. The first frozen/thawed embryo registered by AQHA was in 2012 and the numbers have only recently increased to 300-400 per year. Since in vivo derived embryos do not survive freezing and thawing very well (unless smaller than 300 microns), the vast majority of frozen embryos are derived by intracytoplasmic sperm injection (ICSI) and vitrified. Recently, ICSI has become accepted as a technique for in vitro embryo production and has been embraced by breeders, however AQHA does not currently collect data on the number of foals produced by ICSI. The first registered foal from cooled transported semen was in 1997 and the first from frozen semen occurred in 2000. The use of cooled transported semen increased dramatically to where about 14,000 foals are registered each year from the use of transported semen. It continues to be the procedure of choice, but the numbers of foals from frozen semen have increased over the past 15 years to where now 32% of foals from transported semen were sired by frozen semen. Through education and improvement in cryopreservation techniques and breeding strategies, frozen semen has been more widely accepted. AQHA has set restrictions on the use of certain technologies. For example, for stallions born after 2015 there is a limit on the use of frozen semen beyond 2 years following death.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105300"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386769","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":"Is the sperm-binding test to the hen's egg perivitelline membrane reliable for the functional assessment of equine and donkey semen?","authors":"C.S. Fontes ,&nbsp;H.D.M. Garcia ,&nbsp;M.S. Freitas ,&nbsp;S.C. Teixeira ,&nbsp;E.R. Silva-Junior ,&nbsp;L.L. Oliveira ,&nbsp;T.A.R. Paula ,&nbsp;Y.M.F. Sancler-Silva","doi":"10.1016/j.jevs.2024.105285","DOIUrl":"10.1016/j.jevs.2024.105285","url":null,"abstract":"<div><div>The evaluation of male fertility is traditionally conducted <em>in vivo</em>, but this method is costly and influenced by various factors, such as females and management. The sperm-binding test to the hen's egg perivitelline membrane (HEPM) is an affordable <em>in vitro</em> alternative with promising results in various species, although its application in equids is still not well studied. Thus, this study aimed to: i) assess the feasibility of the sperm-binding test to HEPM at two insemination doses (IDs) for equine and donkey semen; ii) evaluate the influence of seminal plasma and semen cryopreservation on the <em>in vitro</em> sperm-binding capacity to HEPM from both species. In Experiment I, the sperm-binding capacity to HEPM of equid semen with sperm viabilities (SVs) of 0%, 25%, 50%, 75%, and 100% was compared at insemination doses of 10 million (ID10) and 50 million (ID50) total sperm per apparatus containing the membrane. In Experiment II, the sperm-binding capacity to HEPM of fresh and frozen-thawed semen, with or without seminal plasma, was compared. All mean comparisons were evaluated using the PROC GLIMMIX package, considering a statistical difference when P&lt;0.05. The sperm-binding capacity to HEPM was influenced by the insemination dose and the species, being higher for donkeys and for the ID50 dose. When the species was disregarded, the test allowed differentiation of binding potential between infertile seminal samples (0% SV) and samples that exhibited fertile potential (≥25% SV). However, it did not allow differentiation of binding potential between samples with lower and higher SVs (25%, 50%, and 75%). Additionally, cryopreservation and seminal plasma did not influence the sperm-binding capacity to HEPM. In conclusion, this study demonstrated that the sperm-binding test to HEPM can be used to assess the <em>in vitro</em> potential of equid sperm binding capacity. Further studies regarding insemination dose, incubation time of the inseminated HEPM, and the number of repetitions per seminal sample are required to achieve greater accuracy of the test.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105285"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386845","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":"Pharmacologically induced ex copula ejaculation in stallions: assessment of new combined protocols","authors":"J. Garza ,&nbsp;Samaria Del Angel ,&nbsp;M.F. Gallelli ,&nbsp;M. Miragaya","doi":"10.1016/j.jevs.2024.105288","DOIUrl":"10.1016/j.jevs.2024.105288","url":null,"abstract":"&lt;div&gt;&lt;div&gt;Pharmacologically induced ejaculation is an alternative for semen collection in stallions with different physical or behavioral pathologies affecting mounting or copulation. Several protocols have been studied with varying results, so this is still an active research topic. The aim of this study was assessing different protocols to induce ex copula ejaculation. Healthy stallions (n=8, mean age: 8.7 years, range: 5-22 years) were included. Intervals of two days were left between treatments. The following protocols were performed without sexual prestimulation and administering drugs at the same time: xylazine (0.33mg/kg)+butorphanol (0.02mg/kg) (XB), detomidine (0.01mg/kg)+butorphanol (0.02mg/kg) (DB), detomidine (0.01mg/kg)+butorphanol (0.02mg/kg)+oxytocin (20 IU) (DBO) and xylazine (0.33mg/kg)+detomidine (0.01mg/kg)+butorphanol (0.02mg/kg)+oxytocin (20 IU) (XDBO). Other two protocols were imipramine oral administration (2.5mg/kg) followed by xylazine (0.33mg/kg) +butorphanol (0.02mg/kg) (IXB) or detomidine (0.01mg/kg)+butorphanol (0.02mg/kg)+oxytocin (20 IU) (IDBO) 2 hours later. Besides, XB and DBO were also performed with sexual prestimulation (RXB and RDBO) by 10 min of teasing with an estrous mare. Then, stallions were left in their boxes for 5 min, before drug administration. Semen was collected into a non-spermicidal plastic bag. Interval from treatment to ejaculation (min.), volume (ml) and concentration (sperm x10&lt;sup&gt;6&lt;/sup&gt;/ml) of the ejaculate were compared between treatments by one-way ANOVA followed by Bonferroni test. The percentage of stallions that ejaculated post-treatment were compared between groups by Chi-Square test. Differences were considered significant with P&lt;0.05. Values are expressed as mean ± SD (Graph Pad). Ejaculation was observed in 37.5% (3/8) of stallions of XB and RXB, in 25% (2/8) of individuals of DBO and RDBO, in 50% of IDBO (4/8) and in 62.5% (5/8) of IXB. Any stallion of DB or XDBO group ejaculated. Effectiveness for triggering ejaculation was not significantly different between treatments (P=0.06). The mean interval from treatment to ejaculation (min) was 2.01±0.9, 1.7±0.5,1.9±0.7, 1.4±0.14, 1.8±0.4 and 1.9±0.4 for XB, RXB, DBO, RDBO, IXB and IDBO, respectively, without significant differences between groups. The mean ejaculate volume was greater in stallions of RXB (31.7±8.5 ml) than in all other protocols (P˂0.01), except for RDBO (13.8±3.8ml). Mean ejaculate concentration was greater in individuals of DBO (723 ± 5 sperm x106/ml), IDBO (672±62 sperm x106/ml) and IXB (786±56 sperm x106/ml), than in those of RXB (262 ± 170 sperm x106/ml) (P˂0.01). In conclusion, except for DB and XDBO, all protocols were effective in triggering ejaculation in some stallions. Sexual prestimulation only increased the ejaculate`s volume in stallions treated with xylazine+ butorphanol. As not all individuals ejaculate with all protocols, several trials using different treatments should be performed in case of failu","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105288"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386859","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}