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Myeloperoxidase in Equine Semen: Concentration and Localization during Freezing Processing 总被引:1,自引:0,他引:1
Jérôme Ponthier Maud DesvalsThierry Franck PhD Geoffroy de la Rebière Marc SpalartEric Palmer Didier Serteyn Stéfan Deleuze 《Journal of Equine Veterinary Science》2012,32(1):32-37
Myeloperoxidase (MPO) is a pro-oxidant enzyme contained in and released by neutrophils during degranulation or after lysis. Post-thaw semen contains MPO, and concentration of this enzyme is associated with decreased motility. The aim of this study was to determine kinetics of MPO concentration during freezing, its origin, and its impact on frozen-thawed semen. Forty ejaculates were used. Semen was frozen using the classical freezing procedure. MPO concentrations were assayed in fresh semen, after centrifugation, and after cooling down to 4°C. Post-thaw MPO assay results and spermogram characteristics were determined. MPO immunocytochemistry was performed on 4 different ejaculates at each step of freezing procedure. MPO concentration increased after cooling down to 4°C and thawing compared with fresh semen. As temperature decreased, MPO was higher or tended to be higher in post-thaw poor quality samples. Nonsperm cells showed various degrees of MPO immunostaining and were observed as epithelial cells with nuclear pyknosis and keratinization. MPO immunostaining increased in medium and decreased in nonsperm cells during freezing. Our study shows that MPO concentration in equine semen increases when temperature decreases. We hypothesize that nonsperm cells present in fresh semen could release MPO. 相似文献
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Contents: At the Bundesanstalt Wels tests with frozen-thawed boar semen in plastic straws were conducted. The influences of straw volume, method of freezing and thawing extender were investigated. The straw volumes of 1, 0 and 0, 5 ml showed significantly better thawing results in a preliminary trial than the 5, 0 ml Macrotub. When semen was frozen in a computerized freezer with automatic seeding, all tested straw. volumes gave significantly better results than the straws frozen in static N2-vapor. A ready to use commercial extender was used for thawing with good results to simplify the handling of deep frozen semen for on farm insemination. In the main trial 60 ejaculates from 10 boars were frozen in 1, 0 ml straws in the computerized freezer. Three in vitro parameters for fresh semen (motility, osmotic resistance and keeping quality under standard conditions) and two parameters for thawed semen (motility and percentage of sperm with normal apical ridge) were recorded and correlated. The osmotic resistance test proved to be well suited as means of predicting the fitness of an ejaculate for deep freezing, but the other two fresh semen parameters showed poor correlation with the parameters for thawed semen. From the parameters for thawed semen a freezing score was derived as a measure of the freezability of single ejaculates and boars. A preliminary insemination trial gave satisfying farrowing rates. Inhalt: Zur Tiefgefrierung von Ebersamen: Untersuchungen zum Einfluß von verschiedenen “Straw”-Volumina, Einfriewerfahren und Auftaubedingungen Ander Bundesanstalt Wels wurden Tiefgefrierversuche mit Eberspermain Kunststoffpailletten durchgeführt. Die jeweiligen Einflüsse von Paillettenvolumen, Gefrierverfahren und Auftauverdünner wurden untersucht. In Vorversuchen erwiesen sich die Paillettenvolumina 1, 00 und 0, 5 ml gegenüber den 5,0 ml Makrotüb hinsichtlich Kopfkappenintegrität und Auftaumotilität signifkant überlegen. Die Auftauergebnisse der im computergesteuerten Freezer mit automatischem Seeding eingefrorenen Proben waren für alle untersuchten Paillettengröβen signifikant bis hoch signifikant besser, als die Werte der im statischen Stickstoffdampf eingefrorenen Proben. Als Auftauverdünner wurde ein handelsüblicher Fertigverdünner gewählt um eine besonders für den Eigenbestandsbesamer wichtige einfache Handhabung der TG-Besamung zu gewährleisten. Im Hauptversuch wurden 60 Ejakulate von 10 Ebern verwendet und in 1, 0 ml Pailletten im programmierbaren Freezer eingefroren. Drei Frischsamenparameter (Motilität, osmotische Resistenz und Haltbarkeit unter Laborbedingungen) und zwei Auftausamenparameter (Motilität und Kopfkappenintegrität) wurden erhoben und korreliert. Dabei enwies sichder osmotische Resistemtest als für die Beurteilung von Frischsperma hinsichtlich TG-Eignung gut geeignet, während die beiden anderen Frischsamenparameter keine bzw. nur schwache Korrelation zu den Auftauergebnissen aufwiesen. Aus den Auftausamenparametern wurde eine “Einfrierbarkeitszahl” als Maβ für die Eignung des Spermas zur Tiefgefrierkonservierung der einzelnen Eber bzw. Ejakulate erstellt. Dabei enwies sich der osmotische Resistenztest als für die Beurteilung von Frischsperma hinsichtlich TG-Eignung gut geeignet. Bei einem orientierenden Besamungsversuch wurden zufriedenstellende Abferkelraten erzielt. 相似文献
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AB Caselles A Miro‐Moran A Morillo Rodriguez JM Gallardo Bolaños C Ortega‐Ferrusola GM Salido FJ Peña JA Tapia IM Aparicio 《Reproduction in domestic animals》2014,49(2):254-262
Apoptosis in the testis is required to ensure an efficient spermatogenesis. However, sometimes, defective germ cells that are marked for elimination during this process escape elimination in the testes, giving rise to ejaculates with increased percentages of abnormal and apoptotic spermatozoa and a high percentage of apoptotic bodies. Apoptosis markers in the ejaculate have been associated with low fertility, either in animals or humans. Therefore, the goal of this study was to investigate whether fresh equine semen contains apoptotic bodies [initially named Merocyanine 540 (M540) bodies] and to study the relationship between the quantity of these bodies and cell concentration, the volume of ejaculate, viability and motility. Moreover, we also studied whether the presence apoptotic bodies in fresh semen was related to the resistance of the stallion spermatozoa to being incubated at 37°C or being frozen and thawed. Fresh equine semen was stained with fluorescent dyes such as M540 and Annexin‐V. Active Caspase 3 was studied in fresh semen through Western blotting and immunofluorescence with a specific antibody. Sperm kinematics was assessed in fresh, incubated and thawed samples using computer‐assisted semen analysis, and viability was evaluated with the LIVE/DEAD Sperm Viability Kit. Overall, our results demonstrate for the first time the presence of apoptotic bodies in equine semen. The quantity of apoptotic bodies was highly variable among stallions and was positively correlated with Caspase 3 activity in fresh samples and negatively correlated with the viability and motility of stallion spermatozoa after the cryopreservation process. 相似文献
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The Use of Cefquinome in Equine Semen Extender 总被引:1,自引:0,他引:1
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. 相似文献
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猪精液冷冻技术的研究 总被引:31,自引:0,他引:31
以解冻后的精子活率、活力、质膜完整率、顶体完整率和人工授精结果为判定指标 ,比较了几种冷冻稀释液、解冻液及冷冻 -解冻程序对猪精液冷冻的效果 ,并对所筛选出的最佳稀释液和冷冻程序做了改进。结果如下 :(1) 号稀释液冷冻解冻后的精子活力 (32 .4± 7.3) %、活率 (4 2 .2± 3.2 ) %、顶体完整率 (6 2 .3± 0 .8) %和弯尾率 (4 2 .6± 7.5 ) %均显著高于 、 、 号稀释液 (P<0 .0 5 )。 (2 )在 号稀释液中添加 0 .5 %、1%和 2 %的 O.E.P(氨基 -钠 -十二烷硫酸酯 )使精子活率提高 5 %、弯尾率提高 6 %、顶体完整率提高近 10 % ,与对照组相比差异显著 (P<0 .0 5 )。 (3)应用改进的稀释液 ( 液加 1% O.E.P) ,细管法比颗粒法冷冻精子活率提高近 7个百分点 ,活力提高 6个百分点 (P<0 .0 5 )。(4 )室温预平衡 4h的精子活力和活率都比对照组 (0 h)有显著提高 (P<0 .0 5 ) ,而预平衡 2、6 h的精子活力和活率与对照组差异不显著 (P>0 .0 5 )。 (5 ) 号解冻液解冻后精子活力 (4 2 .9± 2 .6 ) %显著高于 、 、 号解冻液 (P<0 .0 5 )。 (6 )用 5 0℃水浴解冻精子的活力 (4 6 .3± 2 .6 ) %显著高于 39℃ (4 2 .9± 2 .6 ) %和 70℃水浴 (4 1.1± 5 .7) %解冻的精子活力 (P<0 .0 5 )。 (7)精清和 号解冻液 相似文献
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大额牛冷冻精液技术的研究 总被引:4,自引:0,他引:4
本文对大额牛的射精量、精子密度、精子活力等指标进行了测定,同时对大额牛精液的冷冻稀释液、制作程序进行了较为系统的研究。结果得出,大额牛在人工饲养条件下,公牛的射精量为4.92±1.50mL、精子密度为16.79±4.5亿/mL、原精活力0.63±0.078;冷冻精液解冻活力平均可达0.3875±0.0426、冻精解冻后顶体完整率平均52.74%±5.88%,畸形率为12.95%±3.18%,精子复苏率为59.63%±3.67%,其他指标与一般黄牛基本一致。 相似文献
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为加强濒危珍稀动物种质资源的保护,开展了濒危珍稀禽类—红腹锦鸡的人工授精研究。2005年5月26日~6月8日,对8只红腹锦鸡用手按摩其背、尾部采精12次。初测其精液品质,结果表明,采精量平均(0.114±0.016)mL(0.01~0.2 mL),每毫升精液中精子平均3.2亿个(3.0~3.3亿),活力9级以上,pH值6.5,偏酸性,淡乳白色(半透明似冲熟的藕粉),微腥。鲜精分别加11%蔗糖—卵黄稀释液(3号液);11%蔗糖—0.1%柠檬酸三钠—卵黄稀释液(5号液);5%葡萄糖—卵黄稀释液(8号液)。精子活力达8~9级。用3%柠檬酸三钠—卵黄稀释液(2号液)稀释,精子活力2级。冷冻时,用11%蔗糖溶液100 mL中加16 mL鲜卵黄,加5 mL甘油,配制的3号冷冻稀释液稀释,解冻后,精子活力达4级。优于试验中选拟的其它配方(加入5 mL甘油的5号冷冻液,解冻后精子活力为3级;加入5 mL甘油的8号冷冻液,解冻后未见活的精子)。如果冷冻稀释液中甘油改为6 mL,则解冻后精子全部死亡。 相似文献