Journal of Equine Veterinary Science最新文献

{"title":"Donkey sperm activating potential in heterologous ICSI","authors":"J. Otero ,&nbsp;D. Neild ,&nbsp;S.M. Gaviria ,&nbsp;A. Camporino ,&nbsp;S. Morado ,&nbsp;C.B. Castex ,&nbsp;P. Cetica ,&nbsp;C. Arraztoa","doi":"10.1016/j.jevs.2024.105311","DOIUrl":"10.1016/j.jevs.2024.105311","url":null,"abstract":"<div><div>There is a growing interest in the preservation and production of donkeys and mules, both species of significant importance in Argentina. The aim of this study was to evaluate the ability of cryopreserved donkey sperm to activate and induce pronuclear formation when injected into heterologous oocytes, as a potential predictor of future seminal capacity to produce embryos in vitro. Porcine cumulus-oocyte complexes (COCs) were obtained from ovaries of slaughtered gilts, selected, and then incubated, for 44 hours at 39°C, with 5% CO2 and 100% humidity, in 199 medium supplemented with 50 μg/ml gentamicin sulfate, 10% (v/v) porcine follicular fluid, 0.57 mM cysteine, 2 IU/ml equine chorionic gonadotropin, and 10 ng/ml epidermal growth factor. The matured COCs were denuded using serial passages through glass Pasteur pipettes. Donkey and equine sperm, cryopreserved with an egg yolk-free extender using a slow-freezing curve in an automatic freezer, were thawed in a water bath at 37°C for 1 minute and selected using Androcoll-ETM. A small aliquot was placed in a drop of 10% (v/v) polyvinylpyrrolidone in modified Whitten's (MW) injection medium. The oocytes were each placed in a drop of MW and intracytoplasmic sperm injection (ICSI) was performed. Porcine oocytes were injected with either equine sperm (control group) or donkey sperm (experimental group). As a control for the ICSI technique, and to assess parthenogenetic activation, mature oocytes were injected without depositing any sperm into the cytoplasm (Sham). Presumed zygotes were cultured for 16-18 hours in NCSU-23 medium at 39°C, with 5% CO2 and 100% humidity. They were then fixed for 15 minutes in 0.5% (v/v) glutaraldehyde in phosphate-buffered solution and stained for 10 minutes with Hoechst 33342 (5 µl/ml). Finally, they were classified as: 1) Activated: two pronuclei (2-PN) or one pronucleus with the presence of a decondensed sperm and 2) Non-activated: one pronucleus with the presence of a condensed sperm or no evidence of pronuclei. No significant difference was observed (p= 0.73) between activated porcine oocytes injected with donkey sperm: 66.67% (52/78) and those injected with equine sperm: 62.86% (44/70). Parthenogenetic development in the Sham group was 4.6% (3/65). Therefore, similar to equines, heterologous ICSI using donkey sperm could predict the future capacity of donkey semen for in vitro embryo production.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105311"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386864","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 role of different extenders in pre-centrifugation for optimizing equine semen cryopreservation","authors":"B.M. de Zutter ,&nbsp;C.P. Freitas-Dell'Aqua ,&nbsp;J.A. Dell'Aqua Jr ,&nbsp;C.M. Trinque ,&nbsp;M.S. Frasson ,&nbsp;L.E.F. Canuto ,&nbsp;T. Troncarelli ,&nbsp;G.A. Monteiro ,&nbsp;M.A. Alvarenga ,&nbsp;F.O. Papa","doi":"10.1016/j.jevs.2024.105339","DOIUrl":"10.1016/j.jevs.2024.105339","url":null,"abstract":"<div><div>Cryopreservation can cause injuries to sperm, thus the inclusion of extenders is crucial in maintaining viability outside the reproductive tract. This study evaluated the effectiveness of 4 extenders: Skim Milk (G1), Skim Milk + Pentoxifylline (G2), Skim Milk + Cholesterol (G3), and Cholesterol + Casein (G4) in maintaining the kinetics and integrity of the cytoplasmic membrane of equine spermatozoa. Semen was collected from 4 stallions with a total of 28 ejaculates. Each ejaculate was divided into 4 aliquots, the samples were cryopreserved and thawed (Papa et al. 2008; Animal Reprod Sci 107:293-301). The evaluation of sperm kinetics, including total motility (TM), progressive motility (PM), percentage of sperm with rapid movement (RAP), and average path velocity (VAP), was performed using CASA (HTM-IVOS 12 Hamilton Thorne Research, Beverly, MA, USA). The integrity of the cytoplasmic membrane (IM) was evaluated using the technique of Harrison and Vickers (Reproduction. 1990; 88:342-352). Statistical analysis was performed using the Kolmogorov-Smirnov test, ANOVA, and Tukey, with P&lt;0.05. After cryopreservation and thawing, the groups G3 (78.1 ± 1.5) and G4 (78.9 ± 1.3) showed higher total motility compared to G2 (73.9 ± 1.7). Progressive motility, percentage of rapid sperm, and plasma membrane integrity were higher in groups G3 (44.7 ± 2.3 and 64 ± 2.8) and G4 (45.1 ± 2.3 and 65 ± 2.5) compared to groups G1 (40.8 ± 2.6 and 60.1 ± 3.2) and G2 (41.6 ± 2.6 and 61.2 ± 2.8). The presence of cholesterol in the extenders G3 and G4 was essential for the preservation of membrane fluidity and stability. Cholesterol helps prevent membrane damage during cryopreservation, while the sodium caseinate present in G4 also contributes to the protection of sperm cells. On the other hand, the extender containing pentoxifylline (G2), which functions by inhibiting phosphodiesterase enzymatic activity, leading to an increase in intracellular cAMP, showed worse performance post-thawing, possibly due to rapid depletion of the energy substrate, generating an increase in ATP production, which can lead to cell death. The findings of this study indicate that extenders with the inclusion of cholesterol (G3 and G4) are more effective in cryopreservation of equine semen than extenders based solely on skim milk. It is known that kinetic parameters of total motility and membrane integrity have proven to be crucial components in fertility prognosis, as these characteristics are interrelated (Foster et al. Theriogenology. 2011; 75:1499-1505). Therefore, the incorporation of cholesterol into the sperm membrane is an effective strategy to increase the viability of cryopreserved semen, particularly in stallions with low resistance to thermal shock, ensuring better results in equine reproduction programs.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105339"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386972","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":"Histopathological and diagnostic aspects of glanders based on a case series from Brazil","authors":"A.F.C. Nassar ,&nbsp;D.P. Chiebao ,&nbsp;C. Del Fava ,&nbsp;S. Miyashiro ,&nbsp;V. Castro ,&nbsp;R.A. Ogata ,&nbsp;J.M. Yamamora ,&nbsp;C.A.S. Monteiro ,&nbsp;E.J.B. Monteiro","doi":"10.1016/j.jevs.2024.105248","DOIUrl":"10.1016/j.jevs.2024.105248","url":null,"abstract":"<div><div>Glanders is a zoonotic disease of equids caused by the bacterium <em>Burkholderia mallei</em>, responsible for considerable economic loss. This study aimed to describe the clinical manifestations, pathological findings, and also bacteriological and molecular methods for agent detection in naturally infected animals (16 adult horses and one fetus) detected by serological survey from three glanders outbreaks. Of the 16 horses, 6 (37.5%) did not show clinical signs. After necropsy,samples of organs, lymph nodes, lesions and secretions were collectedfor histopathology, bacterial isolation, and PCR. The clinical and gross alterations mainly comprised nasal and respiratory forms in the three outbreaks, and less of the cutaneous form. All tested animals were positive by PCR (100%, n=17) on at least one sample. Of 121 samples analyzed 8.2% (10/121) isolated <em>B. mallei</em> and 41.3% (50/121) were polymerase chain reaction (PCR) positive. This work highlights the importance of combining diagnostic techniques, such as histopathology microbiological culture and PCR,to confirm cases and characterize the morbidity of glanders as well as considering seropositive animals without glanders clinical signs as potential carrier animals affecting disease control programmes.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105248"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142807130","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":"Ethylene glycol toxicosis in 39 sport horses following ingestion of contaminated water: A case report","authors":"Zs. Daradics ,&nbsp;D. Bungărdean ,&nbsp;A.F. Lupșan ,&nbsp;M. Popescu ,&nbsp;O. Bulmez ,&nbsp;V. Ciulu-Angelescu ,&nbsp;V.F. Chelaru ,&nbsp;I. Morar ,&nbsp;M. Mircean ,&nbsp;C. Cătoi ,&nbsp;M.A. Tripon ,&nbsp;C.M. Crecan","doi":"10.1016/j.jevs.2025.105343","DOIUrl":"10.1016/j.jevs.2025.105343","url":null,"abstract":"<div><div>Ethylene glycol toxicosis is a common occurrence in dogs and cats but has been reported in other species as well. Up to date, only one case of ethylene glycol toxicosis has been described in horses, and specific guidelines for treating ethylene glycol intoxication in this species are not available. Here we describe the case of 39 sport horses that ingested water contaminated with ethylene glycol. The main clinical sign was apathy, but more severely affected individuals also developed abdominal pain and laminitis. Treatment was initiated around 24 h after the exposure, using ethanol as an antidote, which was administered initially through a nasogastric tube and subsequently via perfusion. Five horses required hospitalization, but all 39 horses recovered. Based on biochemical measurements performed at different timepoints, we proposed several markers that can be used to determine the need for specialized care early on during the treatment.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105343"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142927253","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":"Prolonged dismount latency and its relationship with pregnancy outcomes in Thoroughbred horses","authors":"A. Medica,&nbsp;R. Griffin,&nbsp;A. Swegen,&nbsp;Z. Gibb","doi":"10.1016/j.jevs.2024.105304","DOIUrl":"10.1016/j.jevs.2024.105304","url":null,"abstract":"<div><div>In equine reproductive management, understanding the factors that influence pregnancy outcomes is essential for optimising breeding success. In the Thoroughbred breeding industry, dismount semen (residual post-copulatory urethral semen) is frequently collected and assessed as a proxy for ejaculate quality. Given that stallion ejaculate is fractionated, we hypothesised that the composition of dismount semen is influenced by the duration of time the stallion remains mounted on the mare following ejaculation, and that prolonged post-ejaculatory intromission would allow more of the ejaculate to be deposited into the uterus, potentially improving pregnancy rates. The aim of this study was to investigate the relationship between dismount latency—defined as the time a stallion remains mounted on a mare post-ejaculation—and both dismount sample quality and pregnancy outcomes. Dismount latency was measured as the time from the start of ejaculation to the removal of the penis from the mare. Dismount samples were collected from 62 Thoroughbred stallions and analysed for sperm concentration (NucleoCounter) and motility parameters (iSperm). There was a negative correlation between dismount latency and sperm concentration in the dismount sample (R² = -0.131; P ≤ 0.05). In terms of motility, total motility was negatively correlated with dismount latency (R² = -0.058, P = 0.09), while progressive motility (R² = -0.004, P = 0.67), straightness (R² = 0.041, P = 0.18), and average path velocity (R² = 0.02, P = 0.3) demonstrated no significant correlation with dismount latency. Additionally, a one-tailed t-test revealed that dismount latency was significantly longer when pregnancies were achieved (11.1 ± 0.42 seconds for positive pregnancies vs. 10.2 ± 0.67 seconds for negative pregnancies; P ≤ 0.05). Based on these findings, we conclude that higher-quality dismount samples—those with greater sperm concentrations and motility—are typically obtained when the stallion spends less time on the mare post-ejaculation. Conversely, poorer-quality dismount samples, characterised by lower sperm concentrations and motility, are collected when the stallion spends more time on the mare. This suggests that in cases of prolonged dismount latency, more sperm is being deposited directly into the mare's uterus rather than into the vagina, which is typical of premature dismounts. Thus, prolonged dismount latency appears to improve pregnancy outcomes, and management strategies encouraging longer dismount latency should be considered to enhance breeding success. However, the negative correlation between dismount sample quality and pregnancy success raises concerns about using dismount samples as a reliable indicator of ejaculate quality, highlighting the need for caution when evaluating these samples as proxies for fertility.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105304"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386698","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":"Equine epididymal spermatozoa: A practical guide to collection, processing, and laboratory techniques","authors":"T.M. Tripon ,&nbsp;C.M. Crecan ,&nbsp;Z. Daradics ,&nbsp;I.A. Morar","doi":"10.1016/j.jevs.2024.105328","DOIUrl":"10.1016/j.jevs.2024.105328","url":null,"abstract":"<div><div>Equine epididymal spermatozoa are the last source for gamete rescue in case of sudden death or emergency castration of a valuable stallion. This study aimed to compare the concentration and kinematic parameters of equine epididymal spermatozoa under different protocols. We therefore tested the influence of general and local anesthesia, transport, harvesting technique, refrigeration, seasonality, and age. We hypothesized that all these parameters might influence the quality of spermatozoa and that a standard protocol would be helpful for the practitioner. We analyzed 142 samples after elective orchiectomy using a series of comparative protocols. First, we looked at the effect of general and local anesthesia and compared a TIVA (total intravenous anesthesia) vs PIVA (partial intravenous anesthesia), with or without local lidocaine. We then looked at the influence of transport length and temperature. Once arrived at the laboratory we tested the retrograde flush technique as well as flotation to harvest spermatozoa, we tested two commercial extenders, centrifugation, and refrigeration effect. Finally, we looked at the seasonality and age influence over the study parameters to formulate prognostic characteristics for the owners. All samples collected were analyzed using computer-assisted sperm analysis (SCA^ⓇCASA). The results of this study show that PIVA (partial intravenous anesthesia) has an impact on motility parameters when compared to a TIVA (total intravenous anesthesia) protocol. Local lidocaine did not affect the kinematic characteristics of epididymal spermatozoa. Kinematic characteristics decreased with transport at 5 degrees Celsius but viable spermatozoa were still successfully harvested after 72 hours. The retrograde flush technique was superior for harvesting spermatozoa and centrifugation did not improve any of the parameters tested. Nonetheless, age and season influenced the kinematic characteristics of epididymal spermatozoa. This study looked at healthy stallions that underwent elective orchiectomy therefore the results might indicate more of the characteristic of equine spermatozoa before ejaculation than of epididymal spermatozoa of stallions during emergency castration such as in the case of scrotal hernia or sudden death. However, the results of this study show that epididymal spermatozoa maintain proper viability after testes are removed and that their kinematic characteristics are preserved during general and local anesthesia. During laboratory processing, the retrograde flush technique seems superior, and centrifugation is not necessary. Older stallions are expected to have reduced motility parameters and concentration as well as stallions outside of reproduction season.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105328"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386704","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":"Preliminary results on the use of a novel synthetic extender for artificial insemination in jennies with cooled donkey semen","authors":"L.A. Chapero ,&nbsp;A. Abrego Alvarez ,&nbsp;K.D. Moran ,&nbsp;M.G. Bilbao ,&nbsp;L. Losinno ,&nbsp;L. Rossetto","doi":"10.1016/j.jevs.2024.105274","DOIUrl":"10.1016/j.jevs.2024.105274","url":null,"abstract":"<div><div>Fertility rates using cooled semen in donkeys are acceptable, however in pursuit of an extender that maintains semen viability beyond 48 hours, we tested a new commercially available synthetic extender developed originally for horses (Beyond^Ⓡ, Minitube). This study aimed to evaluate fertility on artificially inseminated jennies with cooled extended donkey semen stored for 72 hours. Semen samples were collected from four Catalan donkeys of proven fertility (three ejaculates each). For the <em>in vitro</em> trial, semen pools containing one ejaculate from each donkey were divided into three samples and randomly assigned to one of the following treatments: 1) Beyond^Ⓡ (B); 2) Beyond^Ⓡ + 2% of centrifuged egg yolk (BEY); and 3) Equiplus^Ⓡ (Minitube) as control (E). Samples from B and BEY were centrifuged at 600 g for 10 minutes, the pellet resuspended to a final concentration of 70 × 10⁶ sperm/ml. The same dilution was used for E. All samples were stored at 5°C. Total motility (TM) and progressive motility (PM) were evaluated at 0, 24, 48, and 72 h using iSperm^Ⓡ (Aidmics Biotechnology). In the <em>in vivo</em> trial, semen from one donkey were treated with BEY or E and stored at 5°C for 72 h, starting with an initial 1 × 10⁹ PM sperm. Healthy and fertile jennies (n=8) were preovulatory inseminated with either BEY or E. Ovulation was induced (200 µg of buserelin; Busereleq, CIMOL Laboratory) at detection of a preovulatory follicle (size 33 ± 1 mm). The interval between AI and ovulation was ± 12 h. Pregnancy diagnosis was performed 14 days post-ovulation. In the <em>in vitro</em> trial, a general linear model with a compound symmetry structure covariance matrix was used. Treatment, time and their interaction were considered fixed effect while semen pool was considered a random effect. P was set at &lt;0.05. For TM, there were no differences at 0 h. For later times, we observed a treatment-by-time interaction effect (P&lt;0.05). For PM, there were no differences at 0 h. For later times, we observed a treatment effect (P = 0.0001) and time effect (P&lt;0.05), but no interaction. At 72 h, the percentual averages for TM and PM were 22.8% and 1.6% for B; 43,2% and 13% for BEY; and 0,6% and 0% for E, respectively. One pregnancy was obtained from the six inseminated jennies with BEY (16.7%), but no pregnancies from the two ones inseminated with E. These preliminary results show that pregnancies can be achieved with cooled semen treated with BEY and stored for 72 h but the low numbers so far do not allow for drawing conclusions.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105274"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386513","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":"Bacterial prevalence in equine semen","authors":"V.L.T. Jesus ,&nbsp;LM.B. Rosa ,&nbsp;M.T.R. Ribeiro ,&nbsp;H. S. da Costa Souza ,&nbsp;A.C. de Oliveira Andrade ,&nbsp;V.S. Freitas ,&nbsp;P.J. Ferraz ,&nbsp;I.J.O. Chávez ,&nbsp;J.C.F. Jacob","doi":"10.1016/j.jevs.2024.105294","DOIUrl":"10.1016/j.jevs.2024.105294","url":null,"abstract":"<div><div>Stallion semen contains a high prevalence of contaminating bacteria from the external genitalia during collection (Rota et al., Theriogenology 2011;76:464-70). These bacteria can induce endometritis in mares predisposed to infections after insemination and can have a deleterious effect on the quality of preserved sperm, affecting their viability and fertility (Samper &amp; Tibary, Theriogenology 2006;66:551-9). In addition, some reports have demonstrated the detrimental effects of some bacteria, such as <em>Pseudomonas aeruginosa</em> or <em>Streptococcus equisimilis</em> on the motility and membrane integrity of equine sperm (Aurich &amp; Spergser, Theriogenology 2007;67:912-8<strong>).</strong> This study aimed to assess bacterial and fungal prevalence in semen from Mangalarga Marchador stallions. Eight stallions were evaluated at random at different stud farms in the state of Rio de Janeiro, Brazil. After washing the penis with water, semen was collected using a Hannover model artificial vagina. A swab was then submerged in the semen, preserved, transported, and refrigerated in Stuart medium until it was sent to the reproduction laboratory. After incubation for 24 hours at 37°C, bacterial plating was carried out on blood agar, MacConkey agar and Chromagar, and fungal plating on Sabouraud medium. After 24 hours of culture, identification was made by morphology and Gram staining. Of the eight stallions evaluated, five (63%) had bacteria in their semen. The bacteria found were <em>Streptococcus</em> spp., <em>Escherichia coli,</em> and <em>Klebsiella</em> spp. (Scholtz et al., Equine Vet J. 2024;1–11). Streptococcus spp. was isolated in 50% of the stallions (4/8), <em>Escherichia coli</em> in 38% (3/8) and <em>Klebsiella</em> sp. in 13 % (1/8) of the samples. Mixed bacterial growth was identified in 38% (3/8) of the samples, two of them were <em>Streptococcus</em> spp. and <em>Escherichia coli</em> and one <em>Streptococcus</em> sp. and <em>Klebsiella</em> sp, respectively. There was no fungal growth in the samples analyzed. Our results are compatible with the bacterial isolates found in a study carried out in Sweden with a total bacterial prevalence of 59%, 53% being <em>Klebsiella pneumoniae</em>, 80% beta-haemolytic <em>Streptococcus</em> and 43% <em>Pseudomonas aeruginosa</em> (Al-Kass, et al, Acta Veterinaria Scandinavica, 2019). It is concluded that bacterial presence in stallion semen should be investigated before it is used in the breeding season.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105294"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386755","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":"Capacitation of frozen-thawed semen improves ICSI-outcomes in horses","authors":"K. Broothaers ,&nbsp;D. Angel-Velez ,&nbsp;T. De Coster ,&nbsp;M. Hedia ,&nbsp;F. Le Gaffric Molto ,&nbsp;K. Smits","doi":"10.1016/j.jevs.2024.105269","DOIUrl":"10.1016/j.jevs.2024.105269","url":null,"abstract":"<div><div>The first report of a repeatable in vitro fertilization (IVF) protocol for fresh semen in horses in 2022 (Felix.et.al., Biology of Reproduction. 2022; 107(6): 1551-1564) paved the way for exploring capacitation applications in equine semen. The effect of equine sperm capacitation prior to intracytoplasmatic sperm injection (ICSI) on subsequent embryo development remains to be determined. In this study, 145 cumulus-oocyte complexes (COC's) were collected from slaughterhouse-derived ovaries and matured for 30-32 hours in TCM-199 with Earl's salts containing 10% (v/v) FBS and 10 µg/mL follicle stimulating hormone and 2 µg/mL luteinizing hormone (Stimufol^Ⓡ, Reprobiol, Ouffet, Belgium). After maturation, the COCs were denuded and the ones showing an extruded polar body were fertilized with piezo-drill ICSI. Half of the mature oocytes (N = 31) were injected with frozen-thawed (FT) semen that was first selected with a sperm separation device (VetCount^Ⓡ Harvester; MotilityCount ApS, Copenhagen, Denmark) and then capacitated in FERT-TALP with penicillamine, hypotaurine and epinephrine (Felix et al., 2022) for 10 hours. The remaining oocytes (N = 33) were subjected to control ICSI with FT-semen that was washed and centrifuged in G-MOPS (Vitrolife, Londerzeel, Belgium). The same batch of frozen-thawed semen was used for both techniques. All presumed zygotes were cultured individually in Global^Ⓡ medium (LGGG-050, LifeGlobal, Guilford, CT, USA) with 10% (v/v) FBS for 5 days, after which they were transferred to DMEM/F-12 containing 10% (v/v) FBS. Cleavage rates were assessed 36h after fertilization, and blastocyst formation was assessed between day 7 and 10 post-ICSI. Statistical analyses were performed using a Fisher's exact test. Cleavage and blastocyst rates were higher for ICSI with capacitated semen (29/31 (93.5%) and 20/31 (64.5%)) than for control ICSI (24/33 (72.7%) and 12/33 (36.4%), respectively) (P = 0.03 and P = 0.02). Capacitation of semen before ICSI could improve commercial ICSI results, but more research is needed to confirm these results and to assess suitability of different stallions for capacitation of FT semen.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105269"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386615","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":"Relationships between metabolism of cryopreserved equine sperm determined by the Seahorse Analyzer and sperm characteristics measured by flow cytometry and computer assisted analysis of motility","authors":"F.M. Straßner ,&nbsp;M. Siuda ,&nbsp;E. Malama ,&nbsp;G. Muffels ,&nbsp;H. Bollwein","doi":"10.1016/j.jevs.2024.105324","DOIUrl":"10.1016/j.jevs.2024.105324","url":null,"abstract":"<div><div>Metabolic activity of sperm has up to now mainly been determined by measuring mitochondrial membrane potential using flow cytometry, but these assays provide little bioenergetic information about sperm metabolism (Moraes et Meyers et al. Anim Reprod Sci. 2018; 194:71–80). Since a few years oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) of cells can be examined using the Seahorse analyzer (Agilent Technologies, Inc., Santa Clara, CA, United States). Studies with this system have already been performed on sperm of humans (Freitas-Martins et al. 2024; Andrology. 2024 12(2):410-421), mice (Balbach et al. Biol Reprod. 2020 103:791-801) and bovine (Moraes et Anim Reprod Sci. Anim Reprod Sci. 2021; 194:71–80), but to our knowledge not in equine sperm. The aims of our studies were to examine intra- and interindividual variabilities of OCR and ECAR in cryopreserved equine sperm and to investigate relationships of these parameters with sperm characteristics determined by flow cytometric assays and computer assisted sperm analysis. Three cryopreserved ejaculates from each of 21 (n=63) warmblood stallions of the Holsteiner Verband (Elmshorn, Germany) were examined. The quality of the semen samples was characterized using different assays during a period of 120 min by measurements every 15 min at a temperature of 37°C. The sperm metabolism was examined using the Seahorse analyzer by measuring basal OCR and ECAR without manipulation performing the Cell Mito Stress. The total motility (TM) of the sperm was assessed by CASA. Furthermore, a flow cytometric assay was carried out to evaluate simultaneously the percentage of sperm with an intact plasma membrane and negative acrosomal status, sperm with a high esterase activity, high mitochondrial membrane potential, low intracellular Ca2+ level (Bucher et al., J Dairy Sci. 2019; 102(12):11652-11669). The SCSA™ was performed to determine the percentage of sperm with a high DNA fragmentation (%DFI) in each sperm sample. There were high variabilities in OCR (34 ± 28 pmol/min of 1 × 106 sperm) as well as in ECAR (1.89 ± 1.18 mpH/min of 1 × 106 sperm) with high inter- and intraindividual variabilities in OCR- and ECAR values within (CVs: OCR: 22.83 - 93.03; ECAR: 29.46 - 90.16) and between (CVs: 65.33; ECAR: 11.39) stallions. There were no high correlations between OCR- and ECAR values and flow cytometric as well as CASA parameters (r-values: min: -0.44; max 0.55). The results of our study show that there are high intra- and interindividual variabilities in oxygen consumption as well as extracellular acidification. As there were no high correlations between the metabolic parameters and other conventionally determined sperm parameters it can be concluded that the Seahorse Analyzer provides new information about sperm quality.</div></div>","PeriodicalId":15798,"journal":{"name":"Journal of Equine Veterinary Science","volume":"145 ","pages":"Article 105324"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143386624","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}