Quality of Fresh,Cooled, and Frozen Semen From Stallions Supplemented with Antioxidants and Fatty Acids

This study assessed the effect of oral supplementation with the primary antioxidants and fatty acids involved in spermatogenesis (L-carnitine, selenium, vitamin E, omega-3, and omega-6) on the seminal quality in fresh, cooled, and frozen semen of stallions (n = 8), using a randomized design. The animals were divided into Group I (n = 4) and Group II (n = 4) for a 30-week experiment. The two groups alternated between nutraceutical supplementation and a placebo over the course of the experiment. Semen collections were performed in two sets: once in the middle of the experiment, before the two groups switched treatments, and once at the end. The volume, appearance, sperm concentration, spermatozoa kinetics, and membrane integrity of fresh semen were evaluated. The spermatozoa kinetics and membrane integrity of cooled (for 24, 36, and 48 hours) and frozen semen were also evaluated. No differences were observed in volume, appearance, and sperm concentration between treatment and control. However, compared with placebo, nutraceutical supplementation increased (P < .05) total motility, trajectory speed, as well as plasma and acrosomal membrane integrity in spermatozoa from fresh semen. In cooled semen, nutraceutical treatment also increased (P < .05) total motility, speed, and membrane integrity of spermatozoa compared with the control. In frozen semen, supplementation increased (P < .05) spermatozoa progressive motility and plasma membrane integrity. Our results suggest a positive, synergistic effect of the antioxidant L-carnitine and selenium on spermatozoa kinetics. Similarly, the increase in plasma and acrosomal membrane integrity could be attributed to higher concentrations of polyunsaturated fatty acids (a key cell-membrane component), combined with the prevention of excess lipid peroxidation by antioxidants. In conclusion, supplementation with nutraceuticals containing fatty acids and antioxidants improved the quality of fresh, cooled, and frozen stallion semen. Therefore, nutraceutical use should increase the success of artificial insemination with cooled and cryopreserved semen.     

四季气温变化对皮埃蒙特牛精液品质的影响

环境温度对精子活力、密度等对有较大程度的影响。本文比较了南阳地区四季气温变化对皮埃蒙特牛精液量、精液色泽、活力,密度等的影响,指出了为保证精液质量在控制环境温度方面应做的工作。     

奶牛冷冻稀释液研究进展

为了提高冻精质量,人们对稀释液进行了大量科学试验,获得许多重要成果.本文综述了奶牛冷冻精液稀释液的研究进展,为进一步研究稀释液和精子之间的相互关系及新型冷冻稀释液的开发和利用提供参考.     

Semen alterations in porcine rubulavirus-infected boars are related to viral excretion and have implications for artificial insemination

Porcine rubulavirus (PoRV), also known as blue eye disease (BED) of swine, causes respiratory and reproductive problems in pigs at several developmental stages. To study the effect of PoRV infection on semen production, five boars were infected with 1 x 10(6) TCID(50)/ml of PoRV strain PAC-3 and evaluated for 59 days post inoculation (DPI). Infected boars developed reproductive tract pathology that included swelling of the testes and epididymides. Analysis of the semen showed that the infection had little effect on semen production in four animals, but semen from one boar showed severe alterations in sperm concentration, motility, and morphology. When motility was analyzed in BTS-diluted semen after 24, 48, or 72 h, alterations were detected in all boars. Furthermore, viral antigen was detected in semen, the seminal plasma fraction, or sperm fraction from all boars. These results showed that PoRV is excreted via semen and, therefore, artificial insemination is a potential route of dissemination.     

Dose-Response of Prolactin to Increasing Doses of the Dopamine Receptor Antagonist,Sulpiride, in Horses: Effect of Season in Mares and Stallions of Estradiol Pretreatment in Geldings

Two experiments were performed to determine whether dopaminergic input to the adenohypophysis (1) differs across seasons in mares and stallions proportionally with changes in prolactin secretion and (2) is altered by estradiol administration in geldings. In experiment 1, prolactin responses to increasing doses of l-sulpiride in eight mares and eight stallions in March, June, September, and December were used to estimate the theoretical dose equivalent to 50% of maximal response. Prolactin areas increased (P < .001) with increasing doses of sulpiride and were greatest (P < .05) in March for stallions, but in June for mares. Mean half-maximal dose, which was assumed to be proportional to the dopaminergic input to the pituitary, was lowest (P < .05) in June and greatest in September. Experiment 2 used the same approach to determine whether the stimulatory effect of estradiol pretreatment on prolactin secretion was associated with an alteration of the half-maximal response. Geldings (n = 6/group) were administered 100 mg of estradiol cypionate in oil, or oil alone, on day 0 (October 3) and increasing doses of l-sulpiride starting on day 6. Estradiol treatment increased (P < .08) the prolactin response to l-sulpiride at 0.41 μg/kg body weight and all higher doses (P < .05); mean half-maximal dose did not differ (P > .1) between groups. We conclude that dopaminergic input to the adenohypophysis of mares and stallions varies with season and that the stimulatory effect of estradiol on prolactin secretion is not associated with a decrease in dopaminergic input to the adenohypophysis.     

植物提取物在家畜精液冷冻保存中的研究与应用进展

植物提取物是从植物中提取的天然抗氧化剂,其在精液冷冻保存过程中能清除多余的活性氧,降低精子的氧化损伤,减少丙二醛产生,提高精子活力与精子各项运动参数,从而提高解冻后的精液品质。本文综述了近几年植物提取物(迷迭香提取物、茶多酚、红景天多糖、葡萄籽原花青素、竹叶黄酮、白藜芦醇、番茄红素和槲皮素)在家畜精液冷冻保存上的应用与研究,旨在为进一步深入研究家畜精子氧化损伤机制提供理论依据。     

广州地区娟姗种公牛精液品质随月份的变化研究

选择在广州地区亚热带气候条件下2岁～6岁的娟姗公牛5头,观察其精液品质(采精量、原精密度、原精活力以及细管精液产量)在全年不同月份的变化情况.结果表明:娟姗公牛的每次采精量和细管精液产量在气温较高的7～9月份会因为采精频率的减少而增加,10月份会因为周采精频率增加而使每次采精量和细管精液产量有明显减少;娟姗公牛的原精密度在气温适宜的2月份和采精频率较低的8月份会出现两个高峰值;娟姗公牛的原精活力在气温较高的月份会出现明显的下降趋势,2月份气温适宜时原精活力最好.     

保护剂在猪精液冷冻保存中的应用

文章综述了猪精液冷冻保存技术的发展简史,阐述了保护剂的作用机理,介绍了常用的保护剂及其应用,以期为猪精液冷冻保存技术的后续研究提供参考,延长猪精液冷冻保存的时间,提高猪人工授精的受精率。     

Quality of Raw and of Cold-Stored Semen in Icelandic Stallions

The aim of the present study was to evaluate the quality of raw and cooled semen in Icelandic stallions. Experiments were performed using seven stallions aged between 3 and 19 years. From each stallion, six ejaculates were collected, and semen quality was determined. Thereafter, the semen was split into eight equal parts and processed with and without centrifugation using the extenders INRA 82-egg yolk, INRA 96, GENT, and Equi-Pro to a final concentration of 30 × 10⁶ sperm/mL. The extended semen was then cooled in an Equitainer, where it was stored for 24 hours, and subsequently refrigerated for another 24 hours at 5°C. Immediately after dilution as well as after 24 and 48 hours storage, sperm motility was analyzed using computer-assisted sperm analyzer, and viability was assessed after dual DNA staining with SYBR-14 in combination with propidium iodide. The results show that the stallion had a significant (P < .05) influence on all variables evaluated in raw semen, and mean (±SEM) values of 43.4 ± 4.3 mL for the volume, 193.0 ± 17.0 × 10⁶ sperm/mL for the concentration, 6.7 ± 0.5 × 10⁹ for total sperm and 73.5 ± 2.1% for total sperm motility, 48.7 ± 2.0% for progressive motility, and 65.3 ± 2.0% for rapid cells were measured. In the cold-stored semen, all variables were significantly (P < .05) influenced by the stallion, extender, and storage time (48 hours). Except for Equi-Pro, all extenders examined were suitable for cooled semen preservation. For storage of more than 24 hours, centrifugation and removal of the seminal plasma were advantageous for all extenders with the exception of Equi-Pro.     

广州地区娟姗公牛与荷斯坦公牛精液品质的比较研究

本试验选择亚热带气候条件下广州地区的娟姗公牛和荷斯坦公牛各5头,比较两个品种公牛的精液品质(采精量、原精密度、原精活力、细管精液产量、冻后活力、低渗膨胀率及穿透率)。研究表明,荷斯坦公牛每次采精的采精量(16.14±0.06 mL)和细管精液产量(189.17±3.11支)都极显著地高于娟姗公牛(4.74±0.05 mL,158.46±2.64支)(P<0.01);娟姗公牛的原精密度(8.95±0.08亿/mL)极显著地高于荷斯坦公牛(8.32±0.07亿/mL;P<0.01);娟姗公牛原精活力(0.731±0.004)高于荷斯坦公牛(0.729±0.003),但两者差异不显著(P<0.05);娟姗公牛精液的冻后活力(0.355±0.003)极显著高于荷斯坦公牛(0.339±0.003;P<0.01);娟姗公牛冷冻精液的低渗膨胀率(34.50%±0.49%)显著高于荷斯坦公牛(31.21%±0.59%;P<0.01);娟姗公牛冷冻精液对去透明带仓鼠卵的穿透率(84.51%±13.83%)显著高于荷斯坦公牛(81.52%±6.13%;P<0.05)。     

猪精液液态保存的研究进展

猪的人工授精主要使用液态保存精液 ,此法保存精子的存活率高、功能受损较小、授精繁殖率较高且操作简便。体外液态保存猪精液时 ,不仅要注意猪精子对温度变化敏感的特性 ,还要考虑酸碱度、离子的种类和强度、渗透压、过氧化损伤和微生物污染等对精子的影响 ,要向稀释液中添加各种保护成分 ,维持精子的存活和功能。目前主要是比较和改进各种稀释液 ,建立准确的精子质量评价方法 ,提高受精率     

The Use of Cefquinome in Equine Semen Extender

Antibiotics are commonly used in equine semen extender for conservation, if semen has to be stored cooled for a maximum of 48 hours or frozen, to eliminate pathogenic or potentially pathogenic bacteria from semen and reduce the risk of postmating endometritis. Little is known about the effect of antibiotics on spermatozoa when semen is stored over a longer period. Cefquinome, a broad spectrum antibiotic and fourth-generation cephalosporin, has been proven to be a powerful drug for the treatment of endometritis and mastitis in different species. Recently in equine studies, it was found to localize in high concentrations in the endometrium. Therefore, cefquinome was used as the antibiotic in semen extender and compared with a commercial semen extender containing gentamicin for effects on motility and membrane integrity of spermatozoa. During the breeding season, ejaculates from nine light horse stallions were collected and half of each ejaculate was stored for 48 hours in modified Kenney type semen extender containing either cefquinome or gentamicin. At 0, 24, and 48 hours, aliquots (20 μL) of the stored semen were evaluated for (progressive) motility and membrane integrity, as well as for various motility parameters by computer assisted sperm analysis. No differences (P > .05) were found in total motility or progressive motility between extenders at any time point. However, there were differences (P < .05) in velocity parameters, although the effect of velocity parameters on fertility is not clear. In general, semen parameters after storage in non-fat dried skim milk semen extender containing cefquinome are comparable with those after storage in semen extender containing gentamicin. The wider spectrum of bactericidal activity possessed by cefquinome may prove to be beneficial in some cases.     

穿山甲、王不留行对卵巢摘除小白鼠乳腺实质发育的影响研究

将Jcl-lcr系18日龄雌性小白鼠28只分为4组,从20日龄时摘除卵巢,28日龄开始,第1组连续给穿山甲、王不留行煎液,每天口服0.2ml(相当于生药0.1g);第2组、第3组、第4组(对照组)以相同方法分别给红花、淫羊藿、益母草煎液和麦芽、莱菔子等煎液及蒸馏水。第45日龄取材检查,结果显示第1组的乳腺分枝数与对照组乳腺分枝相比,差异不显著。同样第1组的乳腺腺泡数与对照组乳腺腺泡数相比,差异也不显著。结论穿山甲、王不留行对摘除卵巢未性成熟小白鼠乳腺发育无作用;穿山甲、王不留行对未性成熟小白鼠乳腺实质发育的促进作用是通过卵巢所致。     

抗氧化剂在绵羊精液保存中的研究与应用进展

人工授精能提高优良种公羊的利用率,提高养羊业的经济效益,也是良种选育和扩繁的必备技术。在此技术中,精液保存是最重要的环节,目前有液态和冷冻保存2种方法。精液在处理和保存过程中会不可避免地产生活性氧(ROS)家族和活性氮(RNS)家族,它们会对精子结构和遗传物质稳定性造成不可逆损伤。因此,在稀释液中添加适量抗氧化剂能够降低ROS和RNS含量,有效延长精子在体外保存时间。本文根据国内外最新进展,综述了抗氧化剂在绵羊精液保存中研究和应用现状,为新型稀释液的研究提供参考。     

奶牛冻精生产中值得注意的问题

作者从原精、冷冻前、冷冻过程中、冷冻后的几个环节论述了质量控制过程中的几个值得注意的问题。     

Influence of Steroidal Anti-Inflammatory Drugs on Viability and Fertility of Equine Semen

Considering the efficiency of steroidal anti-inflammatory drugs (SAIDs) in the immunomodulation of breeding-induced endometritis and the possibility of using these drugs by intrauterine route instead of the parenteral application, the present study aimed at evaluating the effect of SAIDs added to the semen extender on equine sperm viability and fertility. In experiment 1, 15 SAIDs were individually added to a skim milk-based extender and, based on the results of sperm motility, dexamethasone was the drug of choice for the subsequent trials. The effect of dexamethasone on the viability of fresh and 24-hour cooled semen was investigated in experiment 2. In experiment 3, fertility rate was measured in both post-breeding endometritis-resistant and susceptible mares. Although dexamethasone supplementation caused a premature decrease in sperm total and progressive motility and in sperm velocities (P < .05), no difference was observed for sperm membrane integrities and fertility (P > .05). Based on these results, we can conclude that dexamethasone can be added to equine semen at the time of insemination or before cooling, although its use was not able to increase fertility.     

种公牛阴囊周径与部分精液指标之间的相关性研究

通过对山东中大种公牛站现有四个品种(西门塔尔、利木赞、盖普威、荷斯坦)种公牛阴囊周径(scrotal circumference,SC)的测量,分析其与采精量、精子活率、精子密度、6周总采精量、6周总有效精子数5个指标间相关关系.结果表明:不同品种的种公牛SC之间差异显著(P＜0.01);同品种不同年龄的种公牛SC之间差异极显著(P＜0.01);不同品种间种公牛SC与精子活力、精子密度、总有效精子数呈正相关.     

不同采精阶段对后备北极公狐的射精量、精子活力及密度的影响

采用后备北极公狐在配种期内用６个阶段的采精频率检测公狐精液的采出率、射精量、精子活力和密度。精液采出率以３月下旬和４月上旬最高,极显著地高于３月上旬（Ｐ＜００１）;射精量以３月下旬和４月上旬最高,３月上、中旬最低（Ｐ＜００５）;精子活力以４月上旬最高,极显著地高于４月下旬（Ｐ＜００１）,但４月上旬与其他阶段差异不显著（Ｐ＞００５）;精子密度以３月中旬和４月中旬最大,３月下旬最低,二者差异显著（Ｐ＜００５）。结果表明,不同的采精阶段对后备公狐的射精量、精子活力和密度均有一定的影响,尤其对后备公狐发情开始和接近结束阶段的影响更大。     

提高牛细管冻精精子活力的试验研究

本文从细管冻精的3个关键技术环节研究了稀释液的种类对精子活力与存活的影响,确定了细管冻精精子在液氮冷冻度过冰晶期对其损伤最小的最佳熏蒸高度（最佳冷冻温度）和最佳熏蒸时间,并且对成品冷冻的细管冻精不同解冻温度及时间进行研究。由柠檬酸钠和果糖组成的稀释液精子存活时间长,Tris稀释液冷冻后精子活力高于其他配方;牛细管冻精最佳熏蒸距离为2cm,时间影响不显著,7~9min均可;用50℃温水15s解冻精子的活力最高。     

Effects of Repeated Partial Thaw and Refreeze on Post-Thaw Parameters of Stallion Semen Cryopreserved in Cryovials

The current study evaluated post-thaw semen parameters of stallion semen cryopreserved in cryovials and subjected to multiple partial thaw-refreeze cycles. Five fertile stallions were collected twice, and ejaculates were analyzed for concentration, percent membrane integrity, motility, morphology, and sperm chromatin structure (SCSA). Semen processed with freezing extender from each ejaculate was cryopreserved in both 1.2-mL cryovials and 0.5-mL straws. Cryovials were subjected to eight subsequent partial thaw-refreeze cycles. Cryovials were warmed for approximately 30 seconds; then, a sample of cryopreserved semen was removed with a 16-gauge needle, and the cryovial was immediately refrozen in liquid nitrogen. A piece of 0.5-mL straw cut under liquid nitrogen from the same stallion and ejaculate was thawed alongside each cryovial to serve as a control. Thawed samples were analyzed for percent membrane integrity, motility, and SCSA. Post-thaw parameters of motility and membrane integrity were analyzed by one-way or two-way analysis of variance with repeated measures when appropriate. The SCSA data were analyzed using a mixed regression model. Post-thaw motility and percentage of intact sperm were significantly lower when sperm was cryopreserved in cryovials compared to straws. However, these parameters may remain adequate for use in assisted reproductive techniques (ARTs) such as intracytoplasmic sperm injection through all cryovial thaws. Additionally, DNA denaturability was not affected by semen packaging method and was only affected by thaw number, increasing at post-thaws 5 and 6. This technique may offer a unique approach for cryopreservation and utilization of stallion sperm for ARTs in the future.