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.     

The Effect of Trehalose Supplementation of INRA-82 Extender on Quality and Fertility of Cooled and Frozen-Thawed Stallion Spermatozoa

Stallion semen cryopreservation is often associated with poor post-thaw sperm quality. Sugars act as nonpermeating cryoprotectants. The aim of the present study was to evaluate the cryoprotective effect of trehalose on stallion sperm quality and field fertility rates subjected to cooling and freeze–thaw process. Semen samples were collected from six Arabian stallions, divided into five different treatments in a final concentration of 100 × 10⁶ sperm/mL by using INRA-82 extender containing 0, 25, 50, 100, and 200 mM of trehalose then subjected to both cold storage and cryopreservation. Sperm motility, acrosome, plasmatic membrane, and DNA integrity were analyzed, and 57 mares were used to evaluate the field fertility of chilled and frozen-thawed semen. Results showed that the extender containing 100 mM trehalose only increased the functional acrosomal, plasma membrane, and DNA integrities. The inclusion of 50 mM trehalose in semen extender resulted in significantly (P < .05) increased post-thaw total motility compared to the control group, and chilled semen achieved higher pregnancy rates compared to the frozen-thawed one. Pregnancy rate of mares inseminated with frozen-thawed semen (P < .05; 46.15% vs. 36.36%, respectively) was lower than those inseminated with chilled semen (76.47% vs. 68.75%, respectively) but higher than control. In conclusion, addition of 50 mM trehalose yielded the highest quality stallion semen after cooling and post-thawing in terms of motility, integrities of acrosome, membrane, and DNA as well as improved field fertility.     

Stallion Semen Cooling Using Native Phosphocaseinate-based Extender and Sodium Caseinate Cholesterol-loaded Cyclodextrin-based Extender

The objective of this study was to compare semen parameters and embryo recovery rates of cooled stallion semen extended with INRA 96 or BotuSemen Gold. In experiment 1, 45 ejaculates from nine mature stallions were collected, assessed, and equally split between both extenders and then extended to 50 million sperm/mL. Then, the extended semen was stored in three passive cooling containers (Equitainer, Equine Express II, and BotuFlex) for 48 hours. In experiment 2, the same ejaculates extended in experiment 1 were cushion-centrifuged, the supernatant was discarded, and the pellets were resuspended at 100 million sperm/mL with their respective extender. Semen was then cooled and stored as in experiment 1. In both experiments, sperm motility parameters, plasma membrane integrity, and high mitochondrial membrane potential were assessed at 0, 24, and 48 hours post cooling. For experiment 3, 12 mares (n = 24 cycles) were bred with 48 hour–cooled semen from one stallion. Semen was processed as described in experiment 1. Mares had embryo flushing performed by 8-day post-ovulation. In experiment 1, BotuSemen Gold displayed superior total and progressive motility relative to INRA 96 (P < .05). There were no significant differences between the types of containers in any experiment. In experiment 2, INRA 96 and BotuSemen Gold extenders had similar total and progressive motility, but BotuSemen Gold had superior sperm velocity parameters at all timepoints. Embryo recovery was identical for both extenders (50%). Finally, the results obtained herein suggest that BotuSemen Gold is a suitable alternative to be included in semen cooling tests against INRA 96 in clinical practice.     

贵州钙果发展前景、存在的问题及对策

从钙果的生物学特性、适应性、开发利用前景、贵州钙果发展优势等方面进行综合分析,探讨贵州钙果产业发展的对策措施。     

高原型藏羊的采精及细管冻精授配技术试验

为提高青海高原型藏羊选育工作,探索羊的人工采精和细管冻精技术在高寒地区的应用效果,对青海省海北州祁连3只高原型藏种公羊和玛多县30只高原型藏母羊进行了选育试验,实验结果表明:藏种公羊鲜精采集量平均为1.5mL,呈乳白或乳黄色、无气味、有云雾状,平均活力冻前鲜精为0.65,高出一般羊精子标准活力的0.075,冻后为0.35~0.5,高出本省羊精子标准活力0.1,且经冷配后的30只藏母羊均无返情现象。本实验对于藏羊的改良有重要的实践意义,对于改善纯种高原型藏羊也有重要的参考价值。     

浅谈猪精液保存在人工授精中的发展与展望

在世界上主要的猪肉生产国中,猪人工授精(AI)技术的应用率在90%以上。尽管猪冷冻精液技术在不断进步,但由于其效率不高,在猪场实际生产中并不多见使用。在猪人工授精技术中,液态精液是目前应用最广泛的精液,与传统的牛冷冻精液不同,其独特的精子微胶囊保护膜技术,可以对猪精子进行更加完善的液态保存,因此液态精液是目前猪人工授精使用的主要方式。文章综述了猪精液保存技术在人工授精中的应用,并展望了今后的发展方向,为人工授精技术助力恢复生猪生产提供参考。     

早胜牛的采精及细管冻精制作技术要点

近年来，伴随我国畜牧业蓬勃发展的同时，为了更大限度的提高母畜的受胎率和充分利用优秀种畜的基因，人工授精技术得到了不断完善和发展，并在全国各地广泛推广和应用。本文结合实践经验总结出的一些技术措施，阐述了对早胜牛这一地方优良种质资源进行保种和开发利用的重要意义，同时指出人工授精技术和低温冷冻保存精液技术在早胜牛上的实践，对于进一步开发利用培育优质高产肉牛新品种，促进肉牛产业高质量发展具有非常重要的现实意义。通过早胜牛种公牛采精的全过程以及细管冻精制作要点的详细讲解，低温冷冻保存精液技术，可更经济、可靠地实现家畜品种资源的保护，让优良基因长时间保存使得后续研究需要得到保障。     

国家奶牛良种补贴项目在泾阳县实施的成效与启示

为今后奶牛改良工作成果的巩固和推广提供依据,对国家奶牛良种补贴项目在泾阳实施的效果与措施进行总结.结果表明:累计使用良种冷冻精液细管23.7万支,生产改良奶牛3.15万头,且已有1.1万头的优质母牛已正式进入产奶阶段.良种补贴后代奶牛的头胎日产奶量达到27 kg左右,比同期母亲日产奶量高2 kg～3 kg;生奶平均乳...     

Changes in Equine Reproduction: Have They Been Good or Bad for the Horse Industry?

In the past four decades there have been tremendous changes in equine reproduction. Most breeds now allow the use of artificial insemination with fresh, cooled and frozen semen. Artificial insemination has many advantages for the breeder, in particular the control of bacteria through the use of semen extenders containing antibiotics. Deposition of sperm in small volumes onto the uterotubal junction has allowed the use of relatively low numbers of sperm. Intracytoplasmic injection of sperm into oocytes allows older, subfertile stallions to be used as breeding stallions. Advances in mare reproduction have included developing tools for hastening the onset of the breeding season. Other advances include embryo transfer, oocyte collection and transfer, and cloning. The acceptance of reproductive technology depends on the success of the technology, the attitude of the breeders/veterinarians, and the cost/benefit ratio to the industry and breed registry.