共查询到20条相似文献，搜索用时 31 毫秒
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Carlos Ramires Neto Grabriel Augusto Monteiro MSc Rafaela Fatima SoaresCesar Pedrazzi DVM Jose Antonio Dell'aqua Jr. Frederico Ozanan Papa Marco Antonio Alvarenga 《Journal of Equine Veterinary Science》2013
Cooling of equine semen obtained from some stallions results in lower seminal quality and viability when the seminal plasma (SP) is present. The objective of this study was to evaluate the effect of the removal of SP using a Sperm Filter on the viability of cooled stallion semen. For this purpose, 31 stallions were used. Their ejaculates were divided into three groups: CN, semen was diluted with an extender; FLT, SP was removed by filtration; and CT, SP was removed by centrifugation and cooled to 15°C for 24 hours. Sperm kinetics and plasma membrane integrity were evaluated immediately after collection (T0) and after 24 hours of refrigeration (T1). No difference (P > .05) was noted at T1 for total sperm motility (TM), progressive sperm motility, or plasma membrane integrity when semen samples from all the stallions were analyzed. However, when samples from stallions termed “bad coolers” were analyzed (TM = <30% at T1), a difference was observed in TM and progressive sperm motility for CN compared with FLT and CT at T1. Sperm recovery was greater when SP was removed using the filter (FLT) to that when the SP was removed by centrifugation (CN) (89% vs. 81%). Thus, we concluded that filtering with a Sperm Filter is an efficient and practical method for removal of SP from stallion ejaculates, with lower sperm loss than centrifugation. We also found that the presence of SP reduces the quality and viability of cooled semen from stallions whose semen is sensitive to the process of refrigeration. 相似文献
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Classically, evaluation of the breeding stallion for reduced fertility has relied on physical examination of the reproductive system, as well as evaluation of sperm number, motility, and morphology. Over the past 20 years, a number of other diagnostic methods have become available to facilitate reproductive evaluation of the stallion. Specifically, ultrasound imaging has provided much-improved diagnostic methods for evaluation of the external and internal genitalia of the stallion, and these methods have now become routine in evaluation of the stallion. Biochemical analyses of semen can provide useful information for diagnosis of azoospermia (determination of alkaline phosphatase), detection of urine contamination, or changes in pH. Numerous sperm function assays provide information concerning subcellular compartments of the sperm including the plasma membrane, DNA, acrosome, and mitochondria. Data correlating these functional assays with fertility in the stallion are limited in most cases, with the exception of the sperm chromatin structure assay. Finally, the recent sequencing of the equine genome offers the possibility of both marker-assisted selection for fertility traits and more specific information about genetic mutations that may be associated with differing levels of fertility in the stallion. 相似文献
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In this work, we studied retrospectively the influence of several factors on semen quality of the Spanish Purebred (SPB) stallion. Among these factors, we considered stallion age, season, and interval between two consecutive collections. The semen was collected from 11 SPB stallions (634 ejaculates). Semen quality was evaluated based on volume, concentration, and motility of the ejaculates. Results showed that the semen collected in autumn and winter was more concentrated in comparison with that collected in spring and summer, when the volume of ejaculate was the highest. Those ejaculates collected from younger stallions were characterized by having the smallest volume and the highest sperm concentration. When incorporating an SPB stallion into a program of artificial insemination, it is important to take into account all of these factors to achieve the maximum reproductive potential of the animal. 相似文献
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试验旨在研究不同抗氧化剂对马精子低温保存及冷冻效果的影响。选取6匹英纯血种公马作为试验动物,以INRA82液作为基础稀释液（对照组）,分别添加谷氨酰胺（0.015 g/mL）、甘氨酸（0.019 g/mL）、半胱氨酸（0.024 g/mL）、甲硫氨酸（0.015 g/mL）、牛磺酸（0.063 g/mL）、维生素C（0.4 mg/mL）、维生素E（0.5 mg/mL）、褪黑素（0.001 mg/mL）制备成含有不同抗氧化剂的低温保存和冷冻保存稀释液,将浓缩处理后的马精子分别置于上述稀释液中保存或冷冻,检测低温保存48 h后精子运动参数,评价抗氧化剂对精子低温保存的影响;检测精子冻融后运动参数、精子质膜完整性、线粒体膜电势及顶体完整性,评价抗氧化剂对精子冷冻效果的影响。结果表明,精子低温保存48 h后,添加牛磺酸组活精子比例和前向运动精子比例显著高于对照组（P<0.05）,添加维生素C组前向运动精子比例显著高于对照组（P<0.05）。冷冻解冻后结果表明,添加不同抗氧化剂没有改善精子冻融后活精子比例和前向运动精子比例,但添加甲硫氨酸显著延长了精子体外存活能力（P<0.05）;添加不同抗氧化剂精子质膜完整性与对照组间无显著差异（P>0.05）,但甲硫氨酸和甘氨酸组有高于对照组的趋势;添加维生素E和甲硫氨酸组精子线粒体膜电势显著高于对照组（P<0.05）,不同抗氧化剂组精子顶体完整性与对照组间均无显著差异（P>0.05）。结果提示,低温保存时添加牛磺酸和维生素C可以提高精液低温保存效果;冷冻时添加甲硫氨酸能提高精子质膜完整性和线粒体膜电势,且能够延长精子冻融后的存活时间。 相似文献
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Epigallocatechin‐3‐gallate (EGCG) and green tea polyphenols do not improve stallion semen parameters during cooling at 4°C 
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下载免费PDF全文 D Bucci M Spinaci B Mislei B Gadani G Rizzato CC Love C Tamanini G Galeati G Mari 《Reproduction in domestic animals》2017,52(2):270-277
Stallion semen storage for artificial insemination is mainly based on liquid cooled storage. In many stallions this technique maintains sperm quality for an extended period of time (24–72 hr) at 7°C. While this technique is commonly used in the horse industry, there can be a decline in fertility in some stallions, due to an inability of their sperm to tolerate the cool storage process. The aim of the present work was to evaluate the effect of two natural antioxidants (epigallocatechin‐3‐gallate (EGCG) at 20, 60 and 120 μm and green tea polyphenols, and p at .001, .01 and .1 mg/ml) on some sperm parameters (sperm motility, viability/acrosome integrity and DNA quality) in extended semen immediately after its collection (T0) and after 2, 6, 24 and 48 hr of cool storage. Two ejaculates from three trotter stallions were analysed after 48 hr of storage at 4°C. No beneficial effect on the analysed parameters was observed: the two antioxidants were not able to improve sperm quality after 48 hr of storage. These results are in agreement with previous findings on the effect of different antioxidants reported by other researches, who have demonstrated that stallion semen keeps good antioxidant capacity after dilution for 24 hr. In conclusion, the positive effect exerted by antioxidant molecules in other species is not confirmed in the equine one. 相似文献
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XU Wen-hui ZHENG Xin-bao YU Wei-hao MENG Jun LUO Yong-ming LI Hai ZENG Ya-qi WANG Jian-wen WU Zhuang-yuan YAO Xin-kui 《中国畜牧兽医》2017,44(7):2042-2049
This study was aimed to evaluate the effects of various antioxidants, namely glutamine (0.015 g/mL), glycine (0.019 g/mL), cysteine (0.024 g/mL), methionine (0.015 g/mL), taurine (0.063 g/mL), vitamin C (0.4 mg/mL), vitamin E (0.5 mg/mL) and melatonin (0.001 mg/mL) on equine sperm quality after chill or freeze-thaw.Semen were collected from 6 adult thoroughbred stallions, INRA82 was used as the base extender (control group), adding INRA82 with different antioxidants was used in experimental group. Assess the effect of antioxidants on semen by detecting motion parameters after storage at 5℃ for 48 h. Motion parameters, plasma membrane integrity (PMI) and mitochondrial membrane potential were used to evaluate semen quality after thawing. The extender supplemented with 25 mmol/L taurine led to higher TM and PM, and supplemented with 0.4 mg/mL vitamin C obtained significant higher PM compared with control group (P<0.05) after storage at 5℃ for 48 h. The freeze extender supplemented with 0.5 mg/mL vitamin E or 0.015 g/mL methionine significantly increased the mitochondrial membrane potential compare with control group (P<0.05). No significant differences were observed for PMI and acrosomes integrity rate after frozen-thawed (P>0.05), but there was a trend that PMI of adding methionine and glycine group was higher than control group. The results suggested that extender supplemented with taurine and vitamin C could improve the semen preservation effect, and the extender supplemented with methionine could improve plasma membrane integrity, mitochondrial membrane potential of thawing sperm, and also could prolong the survival time of frozen thawed sperm. 相似文献
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Marco Antonio Alvarenga PhD Frederico O. Papa Marcio T. Carmo Tathiane Kievitsbosch Maria Manoela B. Castro Chaves Carlos Ramires Neto 《Journal of Equine Veterinary Science》2012
Removal of excess seminal plasma is sometimes necessary to increase the quality and the longevity of cooled equine semen; moreover, this procedure is an indispensable step aiming to concentrate the sperm cells before freezing equine semen. Typically, the removal of seminal plasma is achieved by centrifugation; however, studies have shown that the force and duration of centrifugation can damage sperm cells and reduce the sperm recovery rate. Recently, new methodologies, such as cushion and filtration, have been described that aim to decrease the mechanical damage of centrifugation to sperm cells. This study aims to compare different methods for concentrating stallion semen. 相似文献
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Taylor D. Stephens Ryan M. Brooks Julia L. Carrington Lulu Cheng Anne C. Carrington Cheryl A. Porr Rebecca K. Splan 《Journal of Equine Veterinary Science》2013
Frozen semen provides several advantages to the breeder relative to fresh or cooled semen. However, some stallions are undesirable candidates for semen freezing due to poor post-thaw motility or longevity caused by membrane damage, osmotic stress, and oxidative stress during cryopreservation. The objective of this study was to determine the effect of post-thaw addition of pentoxifylline, caffeine, or taurine on sperm motility and longevity in equine frozen semen. Pentoxifylline, caffeine, or taurine was incorporated immediately into thawing frozen semen from nine warmblood stallions. Spermatozoa motility and longevity parameters were recorded and analyzed for each additive and for an untreated control. Of the three additives, only pentoxifylline improved total and progressive semen motility relative to that of untreated control. Pentoxifylline also increased semen curvilinear velocity, average path velocity, and straight line velocity relative to those of caffeine, taurine, or control. Semen treated with pentoxifylline also showed greater longevity relative to that of caffeine- or taurine-treated or untreated semen. Taurine improved linearity in comparison to that of semen treated with pentoxifylline, caffeine, or control but did not improve other parameters. Pentoxifylline may be useful in enhancing the quality of equine frozen semen and therefore improving its fertility. Additional studies are warranted that examine the effect of these additives on the conception rate. Pentoxifylline can be used to increase motility and longevity of equine frozen semen and theoretically increase probability of conception and overall breeding success rates. 相似文献
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Influence of partial or total replacement of glycerol by alternative cryoprotectants in Ghent freezing extender on post‐thaw sperm quality in stallions 下载免费PDF全文
下载免费PDF全文 Although glycerol is the cryoprotectant most commonly used in stallions, it has also a considerable toxicity for equine sperm. It was the aim of this study to analyse the quality of frozen‐thawed stallion semen after complete or partial replacement of glycerol in the freezing extender by alternative cryoprotectants. We hypothesized that partial or total replacement of glycerol by cryoprotectants occurring in cold‐resistant frog, insect or plant species results in similar or better semen quality after freezing–thawing. As basic medium, the commercial Ghent basic extender was used and either supplemented with glucose and urea, trehalose and proline, or trehalose and betaine. Based on a series of preliminary experiments, semen was frozen in either commercial Ghent cryopreservation extender (Ghent control), Ghent glucose–urea extender or a Ghent combined extender (glucose–urea, trehalose‐betaine and trehalose‐proline; volume ratio of 2:1:2) in a computer‐controlled rate freezer. After freezing–thawing, semen was analysed for motility, membrane integrity, phosphatidylserine translocation, mitochondrial membrane potential and chromatin condensation. No differences between Ghent control and Ghent glucose–urea extender were seen, while all endpoints except DNA integrity were negatively affected in Ghent combined extender (e.g., progressive motility: Ghent 49.2 ± 3.7, Ghent glucose–urea 46.5 ± 4.6, Ghent combined 24.4 ± 2.8%; p < .001). In conclusion, glycerol concentration in a commercial freezing extender for equine spermatozoa can be successfully reduced when urea as an additive cryoprotectant is added and the glucose concentration is elevated. However, total glycerol replacement with urea, betaine, proline and trehalose was less successful. 相似文献
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人工授精技术是一种较先进的辅助生殖技术,可提高繁殖育种的效益,但因猫类较独特的生殖特点,其人工授精技术发展受限。近年来,科研人员和临床工作者借鉴其他物种相对成熟的人工授精技术,结合猫类生殖特点,对猫人工授精过程中精液采集、精液品质评估、精液保存、刺激母猫排卵方式及具体的人工输精等相关技术进行了优化和创新,如开发了新颖实用的尿导管法采精技术以采集猫精液;在传统精液品质评估的基础上,配合使用荧光染色技术、流式细胞技术及体外受精技术对猫精液品质进行评估;探索不同冷冻液配方及冷冻程序对猫精液保存效果的影响,并尝试使用超快速冷冻法保存公猫遗传物质;研究使用不同种类及不同剂量的激素对诱导母猫发情和排卵效果的差异,以及不同输精部位、方法及时机对母猫妊娠结果的影响等。尽管其中一些新技术与方法已在试验动物中彰显出了其实际应用潜力,但当下依旧还存在操作相对复杂、成本较高、效果低于预期等问题,导致目前猫人工授精技术的应用较局限,不如在犬或其他动物中应用广泛,未来还需更进一步研究和探索才能满足临床实际应用的需要。文章就上述猫人工授精技术中各个方面的研究进展进行了综述,以期为该技术的进一步研究和应用提供参考。 相似文献
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Ziyad Al-Kass Alexander Brown Anders Johannisson Theodoros Ntallaris Jane M. Morrell 《Reproduction in domestic animals》2021,56(6):848-856
Although single layer centrifugation (SLC) selects robust spermatozoa from stallion semen, the effect of individual variation has not been studied in detail. The objective of this study was to determine the variation among stallions in the effects of SLC on sperm quality during cooled storage for up to 48 hr. Semen samples from seven stallions (18 ejaculates) were split, with one portion being used for SLC and the other serving as a control (CON). Sperm quality (kinematics, reactive oxygen species (ROS) production, membrane integrity (MI) and chromatin integrity) were analysed at 0, 24 and 48 hr using computer-assisted sperm analysis and flow cytometry. Sperm quality was better in SLC than in CON at all timepoints, especially chromatin integrity and MI (p < .0001 for both), and some categories of ROS production (e.g. proportion of live hydrogen peroxide negative spermatozoa, p < .0001), but the degree of improvement varied among stallions and type of ROS (p < .05–p < .0001). Total and progressive motility were also better in SLC samples than in CON at 24 and 48 hr (p < .0001), although the effect on sperm kinematics varied. The interaction of treatment, time and stallion was not significant. In conclusion, sperm quality was better in SLC samples than in CON, although there was considerable individual variation among stallions. The improvement in sperm quality, particularly in chromatin integrity, was clearly beneficial, and therefore the use of this technique would be warranted for all stallion semen samples. 相似文献
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REASONS FOR PERFORMING STUDY: A method of removing equine arteritis virus (EAV) from equine semen used for artificial insemination is urgently needed. Recent medical studies suggest that a double semen processing technique of density gradient centrifugation followed by a 'swim-up' can provide virus-free sperm preparations for assisted reproduction. OBJECTIVES: To investigate the use of the double semen processing technique to obtain virus-free sperm preparations from stallion semen containing EAV. METHODS: Aliquots of an ejaculate from an uninfected stallion were spiked with virus and processed by the double processing technique. The sperm preparations were tested by PCR for the presence of EAV. The procedure was repeated using an ejaculate from a known shedding stallion, testing processed and unprocessed aliquots by PCR and virus isolation. RESULTS: Virus-free sperm preparations were obtained using the double sperm processing technique. The 'swim-up' step is apparently required to ensure complete virus removal. CONCLUSIONS: The double semen processing technique is potentially a useful and simple tool for the removal of EAV from the semen of shedding stallions. POTENTIAL RELEVANCE: The inclusion of density gradient centrifugation and 'swim-up' in protocols for the processing of semen for artificial insemination could help prevent the transmission of viral diseases carried in semen, such as EAV. 相似文献
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Influence of Semen Storage and Cryoprotectant on Post-thaw Viability and Fertility of Stallion Spermatozoa 总被引：1，自引：0，他引：1
C.M. Melo MS F.S. Zahn PhD I. Martin MS C. Orlandi MS J.A. Dell'Aqua Jr. PhD M.A. Alvarenga PhD F.O. Papa PhD 《Journal of Equine Veterinary Science》2007,27(4):171-175
The aim of the current study was to verify that stallion spermatozoa could be cooled for 24 hours and then frozen. In experiment I, one ejaculate from each of 13 stallions was used. Semen was collected and split into two parts; one part immediately frozen using standard cryopreservation techniques and the other diluted, stored in an Equitainer for 24 hours, and then frozen. In experiment II, one ejaculate from each of 12 stallions was collected, diluted with Botu-Semen, and split into two parts: one cooled in an Equitainer and the other in Max-Semen Express without prior centrifugation. After 24 hours of cooling, the samples were centrifuged to remove seminal plasma and concentrate the sperm, and resuspended in Botu-Crio® extender containing one of three cryoprotectant treatments (1% glycerol + 4% dimethylformamide, 1% glycerol + 4% dimethylacetamide and 1% glycerol + 4% methylformamide), maintained at 5°C for 20 minutes, then frozen in nitrogen vapor. No difference was observed between the two cooling systems. The association of 1% glycerol and 4% methylformamide provided the best post-thaw progressive motility. For experiment III, two stallions were used for a fertility trial. Forty-three inseminations were performed using 22 mares. No differences were seen in semen parameters and pregnancy rates when comparing the two freezing protocols (conventional and cooled/frozen). Pregnancy rates for conventional and cooled/frozen semen were, respectively, 72.7% and 82.3% (stallion A), and 40.0% and 50.0% (stallion B). We concluded that cooling equine semen for 24 hours before freezing, while maintaining sperm viability and fertility, is possible. 相似文献
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Katila T 《Acta veterinaria Scandinavica》2001,42(2):199-217
The article reviews methods used for in vitro evaluation of sperm, with particular emphasis on frozen-thawed stallion sperm. The techniques, limitations of the methods and correlations with fertility results are discussed. Very few studies have tried to find correlation between fertility of frozen stallion semen and laboratory tests. It is difficult and expensive to inseminate an adequate number of mares to achieve statistically significant differences. Significant, but low correlations have been demonstrated between the foaling rate and subjective motility of sperm incubated for 2 h and 4 h at 37 degrees C and hypoosmotic swelling test after 0 and 3 h of incubation. Significant correlations have been reported between the pregnancy rate and viability of propidium iodide-stained sperm assessed by flow cytometry as well as for glass wool and Sephadex filtration tests. No correlations have been detected between fertility and motility immediately after thawing. In spite of that, motility estimation by light microscope is the most commonly used method to evaluate frozen-thawed stallion sperm. Computer assisted automatic sperm analyzers have replaced light microscopy in research projects, but so far nobody has been able to demonstrate a correlation between fertility of frozen stallion semen and any of the motility parameters obtained by these instruments. 相似文献
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Sperm concentration and sperm membrane intactness (SMI) or viability are two measures of sperm quality that provide important but different information about a stallion's reproductive capability. Sperm concentration is a measure that, by itself, informs little about the reproductive status of either the stallion or the ejaculate. Nevertheless, it is part of the product, along with semen volume, that determines total sperm number. The correct calculation of total sperm number directly affects the number of mares a stallion can breed and therefore, fertility. If either sperm concentration or semen volume is incorrectly measured, both the number of mares that a stallion can breed and the fertility of those breedings are affected. Although considerable between-stallion variation exists, sperm concentration, semen volume and total sperm number tend to be seasonal and vary with ejaculation frequency. 相似文献
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马、驴精液的保存与应用技术发展速度相对较慢,文幸从马、驴精液的采集,处理,稀释,冷冻,解冻,授精等几个方面就目前国内外发展情况进行了概述. 相似文献