黄芪多糖对猪精液冷冻保存效果的影响

为了评价黄芪多糖（Astragalus polysaccharides）对外来公猪精液冷冻保存的影响情况,为猪精液冷冻稀释液配方的改良提供理论依据,试验采集美系长白、大约克与杜洛克3种公猪的精液,用添加不同浓度（0、0.01%、0.02%、0.03%、0.04%、0.05%、0.06%）黄芪多糖的冷冻稀释液稀释,0.25 mL塑料细管分装并冷冻,测定解冻后精子活力、畸形率及顶体完整率,并进行相同浓度3种公猪之间和同种公猪不同浓度之间的比较。结果表明,在相同黄芪多糖浓度下,仅杜洛克公猪冻后精子顶体完整率在不添加黄芪多糖时显著高于大约克公猪（P<0.05）,其余均无显著差异（P>0.05）;每种公猪的最佳黄芪多糖添加浓度均为0.04%,在该浓度下精液冻后质量均显著优于对照组（P<0.05）;各种公猪最佳黄芪多糖添加浓度下的精液冻后质量指标之间均无显著差异（P>0.05）。总之,稀释液中添加0.04%黄芪多糖在长白、大约克与杜洛克3种公猪的精液冷冻都可以取得较好的效果。     

红景天多糖对藏猪精液冻后品质及精子基因组DNA甲基化的影响

在藏猪精液的冷冻液中添加不同质量浓度的红景天多糖(0,2,4,6,8,10mg/L),制成高密度细管冻精,以冷冻解冻后精子活率、精子畸形率、精子顶体完整率和质膜完整率为精液品质的评价指标,筛选最适红景天多糖添加质量浓度;用甲基化荧光定量法检测3组(鲜精组、未添加组、最适添加组)精子基因组DNA甲基化的水平。结果显示:添加红景天多糖质量浓度为6mg/L组的精子活率显著高于其他组(P<0.05),该组精子畸形率、精子顶体完整率、精子质膜完整率也显著好于未添加组(P<0.05);这3组精子基因组的DNA甲基化水平(0.610 5±0.080 0,0.945 7±0.043 7,0.680 2±0.051 0)中,最适添加组虽显著高于鲜精组(P<0.05),但却显著低于未添加组(P<0.05)。结果表明:在藏猪精液冷冻液中添加质量浓度6mg/L的红景天多糖不仅可以明显改善藏猪精液冻后品质,而且可以明显减少冷冻对藏猪精子基因组DNA甲基化水平的影响,这将为提高藏猪精液冷冻保存效果的进一步研究提供参考。     

猪精液冷冻技术研究进展

本文综述了猪冷冻精液的国内外发展简史,对冷冻保存机理进行了简要阐述,并从冷冻保存稀释液、冷冻保护剂及其浓度、诱发结晶和解冻等方面阐述了影响猪冷冻精液的因素,指出了目前在猪精液冷冻理论和实践中的存在问题,并针对这些问题提出了一定的解决办法.     

海藻糖对猪精液冷冻保存效果的影响

在传统的Tris-柠檬酸-葡萄糖稀释液基础上，分别添加25％、50％、75％、100％的海藻糖，研究不同浓度海藻糖对猪精液冷冻后精子质量的影响。结果表明，海藻糖相对于对照TCG稀释液能够显著改善和提高猪精液的冷冻效果，其最佳添加浓度为25％，冷冻-解冻后猪精子活力、活率、线粒体活性、质膜完整性以及顶体完整率均显著提高（P〈0．05），分别达到41．38％、46．34％、44．56％、43．51％和64．09％。海藻糖可以明显抑制精子获能，获能处理前精子获能率仅为3．68％，而获能处理后达到41．82％，有利于促进精子获能。精液稀释液中甘油的适宜添加浓度为2％，海藻糖只有与甘油共同作用，才能在冷冻-解冻过程更加有效地保护精子。猪精子活力、活率、线粒体活性、质膜完整率、顶体完整率等之间存在极显著的正相关关系（P〈0．01），而与获能处理前精子的获能率存在显著的负相关关系（P〈0．05）。     

黄芩多糖对长白猪精液冷冻保存效果的研究

本研究旨在探讨不同浓度的黄芩多糖添加方案对猪冷冻精液保存效果的影响。猪冷冻处理的精液共6组,分别为空白对照组和添加不同浓度黄芩多糖的试验组（在冷冻稀释液中分别添加0.2、0.4、0.6、0.8、1.0g/L黄芩多糖）,对冷冻-复苏的精液进行精子活力及相关运动参数、精子质膜完整性、顶体完整率等指标检测。结果表明：与空白对照组相比,添加0.4 g/L的黄芩多糖对冷冻-复苏后的精子直线速度、曲线速度、平均路径速度、顶体完整率、DNA完整率和精子抗氧化酶活性均有提高（P<0.05）。0.6 g/L黄芩多糖组精液冷冻复苏后精子的运动参数、顶体完整率及抗氧化酶活性与0.4 g/L黄芩多糖组无显著差异。此外,0.8 g/L黄岑多糖组的质膜完整率及直线性高于其他组（P<0.05）。综上所述,添加不同浓度的黄芩多糖均可提高猪冷冻精液品质,其中提高猪冷冻精液保存质量的最适添加量是0.4 g/L。     

猪精液冷冻技术研究

1956年英国人Polge开始研究猪精液的冷冻保存.后来的研究中,利用牛精液冷冻方法冷冻猪精液,解冻后的精子具有活力,但没有受精能力,虽然在1970年之前有几例猪冻精成功受胎的报道,但其结果不能被重复.     

不同抗氧化剂对猪精液冷冻保存效果的影响

在LEY冷冻稀释液基础上分别添加不同浓度维生素C(0、5、10、20、40、60 mmol/mL)、维生素E(0、0.2、0.5、1.0、2.5、5.0 mg/mL)、SOD(0、100、200、400、600 IU/mL)、CAT(0、50、100、200、300 IU/mL)、GSH(0、1、5、10 mmol/mL)等抗氧化剂,检测冷冻-解冻后精子活力、活率、质膜完整率、顶体完整率、平衡后和解冻后精子MDA含量,以观察5种抗氧化剂对猪精液冷冻保存效果的影响。结果表明:在冷冻稀释液中联合添加100 IU/mLSOD和200 IU/mL CAT明显提高冷冻-解冻后精子活力、活率、质膜完整率和顶体完整率(P<0.01)。     

甘草多糖对猪精液低温保存的影响

通过在猪精液低温稀释液中添加不同质量浓度的甘草多糖(0.0 g/L、0.10 g/L、0.30 g/L、0.50g/L),在4℃低温保存下于试验第1天、第3天、第7天,对猪精子的活率、活力、畸形率、顶体完整率、质膜完整率、SOD及MDA等参数进行测定,探讨甘草多糖对猪精液低温保存应用的效果.结果显示,在精子质量评价和抗...     

绿原酸对猪精液冷冻保存效果的影响

为探究绿原酸对猪精液冷冻保存效果的影响,分别在TCG稀释液中添加不同浓度(15、30、50、80和100 pg/mL)的绿原酸,通过测定冷冻-解冻后精子的活率、顶体完整率、质膜完整率、DNA完整率、超氧化歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)活性来判定对保存效果的影响.结果表明,当添...     

Effect of pasteurized egg yolk on the quality of cryopreserved boar semen

Egg yolk (EY, control) is an essential ingredient of diluents for boar semen cryopreservation. Pasteurized egg yolk (PEY) reduces hygienic risks in processing and is easier to standardize. The aim of this study was to evaluate the in vitro effect of PEY (treatment) on frozen-thawed boar semen. In a split-sample approach (n = 13 boars), it could be shown that there is neither an influence (p > .05) on post-thawing motility (PTM: 5, 30 and 120 min) nor on morphologically intact sperm, percentage of acrosome defects and membrane fluidity using a PEY extender compared to the control. Mitochondrial activity (p = .043), membrane integrity (p = .015) and PTM 300 min (p = .023) were slightly affected in the treatment group. Overall, sperm quality was at a high level in both experimental groups. Further studies are needed to determine the impact of PEY on the fertilizing capacity of boar ejaculates.     

The effect of Curcuma longa extracted (curcumin) on the quality of cryopreserved boar semen

The aim of this study was to determine the optimal concentration of curcumin needed for cryopreservation of boar semen. Semen samples (n = 9) were collected from nine Duroc boars which having proven fertility were used for routine artificial insemination. Semen samples were collected and divided into six groups (groups A‐F) according to various concentrations of curcumin in freezing extender (i.e. 0, 0.125, 0.25, 0.50, 0.75 and 1.0 mmol/L, respectively). The semen was frozen by traditional liquid nitrogen vapor method and stored at ?196°C in the liquid nitrogen tank. After storage, frozen semen samples were thawed at 50°C for 12 s and evaluated for progressive motility, viability and acrosome integrity. The present results indicated that the addition of curcumin at 0.25 (group C) or 0.50 mmol/L curcumin (group D) yielded the higher percentage of progressive motility (33.3 and 36.1%, respectively) (P < 0.001). A significantly higher percentage of acrosome integrity was found in groups B (29.7%), C (31.1%) and D (30.2%) than in the other groups (P < 0.01). However, there was no significant difference in percentage of viability among groups. In conclusion, addition to the freezing extender of curcumin during cryopreservation at a concentration of 0.25 or 0.50 mmol/L is the optimal concentration of curcumin for improving the quality (i.e. increased progressive motility and acrosome integrity) of cryopreserved boar semen.     

High-speed centrifugation of extender of freeze-thaw boar semen

In semen cryopreservation, egg yolk is still widely used as a non-penetrating cryoprotectant. Much has been developed in the search for alternatives for this biological product. This work aimed to evaluate the processed egg yolk through ultracentrifugation and/or sonication in the cryopreservation of swine semen. Twenty-seven semen doses were purchased from a commercial boar stud and processed for cryopreservation using egg yolk lactose 11% (control) extender, processed using two different methods: high-speed centrifugation and sonication. Then, they were submitted to freeze-thawing protocol and were assessed for kinematic and cell structural parameters. Samples in which extenders underwent centrifugation had better results in velocity parameters, meanwhile those that only sonication was performed had poorest results in this parameter. The preservation of the membrane and mitochondria structure had better results when the diluent was only centrifuged in comparison with the other treatments. Therefore, centrifugation of extender containing egg yolk is important for better cryopreservation of swine semen.     

Preservation of boar semen: An update

In the pork industry, artificial insemination and the storage of boar semen in liquid at 17°C are routinely applied to optimize the ejaculate and bring about rapid genetic changes that are reflected in the animal protein. Although the results are satisfactory, they are below what occurs with natural mating. It is currently possible to preserve boar semen with storage at 17°C and slow freezing, since to date there is only one study on vitrification, with negative results applicable only in the case of implementing an intracytoplasmic sperm injection. In both methods and due to the sensitivity of boar sperm to osmotic and temperature changes, there is a loss in the quality of the initial sample; however, slow freezing in boar semen has greater deleterious effects on the sample that are reflected in the pregnancy rates and number of live births. Therefore, only 1% of all inseminations are done with frozen semen. The aim of this review is to provide advances and results of studies conducted on the preservation of boar semen, delving more deeply into the critical points that each of the preservation techniques presents, including bacterial contamination, extender components, temperature, ice nucleation, use of additives in extenders and the main deleterious effects on sperm quality.     

Effects of air exposure and agitation on quality of stored boar semen samples

The objective of this study was to investigate the effects of semen volume, air contact inside semen dose tubes, daily agitation of semen doses and extender type on semen quality, thermo-resistance and bacteria growth in extended boar semen doses preserved over 7 days of liquid storage. Ejaculates from 4 proven terminal cross-bred boars were collected using the gloved-hand technique for 4 weeks and used in the 3 × 2 × 2 factorial study. The effects of treatment (CON: 80 ml doses sealed at the top of the tube; 40HIGH: 40 ml doses sealed at top of tube, and 40LOW: 40 ml doses sealed at top of the liquid), agitation (agitated versus not agitated) and extender type (long-term versus short-term) were investigated on semen quality, thermo-resistance and bacteria growth in boar semen doses. The results of the study revealed that motility (p = .031) and viability (p = .041) in 40HIGH were lower than CON. pH (p < .001) was higher in 40HIGH compared with CON and 40LOW. Agitation did not impact motility (p = .581), progressive motility (p = .870), viability (p = .509) or morphology (p = .970), while long-term extender maintained higher motility (p = .002), progressive motility (p = .036), viability (p < .001) and normal acrosome (p < .001) than a short-term extender. VAP (p = .039) of 40HIGH was lower than CON in a thermo-resistance test. Neither treatment (p > .798, .766) nor agitation (p > .396, .476) impacted bacterial growth in this study. In conclusion, air contact negatively impacts boar semen pH and consequently sperm motility. Semen doses prepared with 80 or 40 ml volumes of extended boar semen with minimal air contact in the tubes yield more desirable semen quality and agitating boar semen doses daily does not have negative or positive effects on boar semen quality.     

抗生素对猪精液常温保存效果的影响

采集9头公猪的精液（其中长白4头、大白4头、杜洛克1头）,对精液进行稀释后,分别加入9种抗生素,在18℃保存,测定精子保存时间和精子存活指数.结果表明：磺胺抗菌效果最好,其有效保存时间为4.38 d,精子存活指数为3.14.恩诺沙星、阿奇霉素、林肯霉素的抗菌效果次之,庆大霉素、青链霉素的抗菌效果再次之,新霉素、卡那霉素的抗菌效果最差.     

云南省种公猪精液中乙型脑炎病毒的监测

应用RT-PCR技术开展云南省种公猪精液中乙型脑炎病毒(JEV)感染监测,进而对阳性样品病毒基因扩增产物进行克隆、测序、比对及系统发育分析。从云南省16个地州797份猪精液中检出JEV阳性样品7份,阳性率0.88%。阳性精液样品中的JEV与基因Ⅰ型毒株PrM基因核苷酸序列同源性为97.5%～98.8%,与其他基因型毒株的同源性介于76.9%～89.8%之间,与疫苗毒株(基因Ⅲ型毒株,S19980008)的同源性为89.1%～89.8%。云南省种公猪精液中JEV属于基因Ⅰ型毒株,与人、猪、蚊虫基因Ⅰ型分离毒株遗传关系密切。     

EGTA对猪精液液态保存的影响

猪精液液态保存体系中,钙离子浓度升高将降低精子活力,缩短精子体外保存时间。常用的金属螯合剂EDTA的溶解度较低,且容易发生沉淀,因此改进螯合剂对于改进猪精液保存液配方,提高猪精液液态保存质量十分重要。本研究中我们在精液液态保存液中分别添加不同浓度(1.5,3,6,12 mmol/L)的钙离子螯合剂EGTA,检测其对猪精子的活力、质膜完整性、顶体完整率、获能情况以及受精能力的影响。结果表明,精液保存液中添加EGTA能显著提高猪精子保存质量,其中添加3 mmol/L EGTA效果最好(P<0.05)。与对照组相比,添加3 mmol/L EGTA时,体外受精率和囊胚率显著提高(P<0.05)。因此,精液保存液中添加EGTA有助于提高猪精液液态保存的质量。     

A preliminary study on the effect of dietary supplementation with cod liver oil on the polyunsaturated fatty acid composition of boar semen

Eight mature Norwegian Landrace boars, of proven fertility and in routine semen production for AI, were fed individually with the same basic diet for 9 weeks. One group of 4 animals served as the control, the remaining 4 boars received a daily supplement of 75 ml cod liver oil (CLO-group). Fifteen consecutive semen samples were collected from each boar. The fatty acid composition of the semen was determined, and the content of the 15 most numerous fatty acids with a chain length longer than 12 carbon atoms was followed over time. In both groups, the proportion of 16:1n-7 decreased significantly, while 16:0 and 22:6n-3 (DHA) increased. By the end of the experiment, DHA had tended to increase and 22:5n-6 to decrease to a greater extent in the CLO-group. A significant difference between the groups was seen for onen-6 PUFA (22:4n-6), which remained unchanged in the control group but decreased in the CLO-group. No change was seen in docosapentaenoic acid (22:5n-3) and eicosapentaenoic acid (20:5n-3) was not found in any sample. These results indicate that CLO supplementation affects the fatty acid composition of boar semen. There were no significant differences in the non-return rates (4–25 days) between the two groups before, during or after the experiment.Abbreviations AA arachidonic acid - AI artificial insemination - area% each fatty acid percentage of the total area of the peaks from gas chromatographic analysis of methylated fatty acids - CLO cod liver oil - DHA docosahexaneoic acid - DPA docosapentaenoic acid - EPA eicosapentaenoic acid - LA linoleic acid - LNA linolenic acid - NR non-return rate - PUFA polyunsaturated fatty acid - T _c the temperature marking the beginning of the phase transition Fatty acid nomenclature: Example, 22:6n-3: 22=number of carbon atoms, 6=number of double bonds,n-3=position of the first double bond counted from the terminate (n or methyl end of the fatty acid chain     

Evaluation of cryopreserved boar semen after supplementation sericin form silkworm (Bombyx mori) in semen extender

Boar cryopreserved semen is scarcely used for artificial insemination due to its quality which is largely reduced by membrane lipid peroxidation. This present study was designed to improve the post‐thawed boar semen quality by determining the optimal level of sericin supplementation (antioxidants) in semen extender. Five levels of sericin supplementation between 0% and 1% (w/v) were examined. Semen was frozen by the liquid nitrogen vapor method, thawed slowly at 5°C for 5 min, and used for the evaluation of sperm quality. The results indicated 0.5%–1% sericin supplementation was more effective on maintenance of sperm viability, acrosome integrity, and mitochondrial functions during freezing–thawing. Moreover, 0.75% sericin supplementation was most protective toward total sperm motility and sperm progressive motility. Additionally, 0.25%–0.75% sericin supplementation significantly suppressed increases in the index of lipid peroxidation. In conclusion, 0.75% sericin is recommended as an alternative component of the freezing extender to improve cryopreserved boar semen. However, further research using AI will be necessary to demonstrate that this indication can be applied to the production of offspring in the farms.