硫酸铜对泥鳅精子活力的影响     

陈玉明  陈杰  王日文  陈凤梅  胡家会 《动物医学进展》2011,32(4):41-44

以泥鳅精子为受试材料,应用计算机辅助精子分析系统,研究硫酸铜(0.1、1、10 mg/L)对泥鳅的精子活力的影响.结果表明,在0 h试验组,0.1 mg/L的硫酸铜明显降低泥鳅精子运动速度,1 mg/L的硫酸铜对运动时间和速度产生显著抑制作用,10 mg/L的硫酸铜时运动百分数、运动时间和速度都产生明显影响(P<0.0...  相似文献   

Delivery of CatSper2 siRNA into Rat Sperms by Electroporation Repressed Ca2＋ Influx During Sperm Hyperactivation     

ZHANG Zhen  ZHOU Xuan  LI  Hui-xia CUI  Qun-wei YU  Jing WANG  Gen-lin 《中国农业科学(英文版)》2011,(12):1958-1967

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AMPK激活剂在绵羊精液冷冻保存中的作用研究     

汪棋  汪长建  魏宗友  陆汉希  姚晓磊  杨花  王锋  张艳丽 《畜牧兽医学报》2020,51(12):3033-3045

旨在研究AMPK激活剂二甲双胍（metformin，Met）和阿卡地新（acadesine，AICAR）对绵羊精液冷冻保存效果的影响。本研究首先在冷冻基础稀释液中分别添加不同浓度（0、100、200、300、400、500 μmol·L^-1）的Met和AICAR，冷冻解冻后根据精子活力、运动性能和结构完整性指标筛选出最佳的添加浓度（400 μmol·L^-1 Met、200 μmol·L^-1 AICAR）；然后分别使用不同的冷冻稀释液（对照组：稀释液；Met组：含400 μmol·L^-1 Met的稀释液；AICAR组：含200 μmol·L^-1 AICAR的稀释液）冷冻精液，解冻后检测精子中AMPK蛋白表达、顶体酶活性、代谢指标、线粒体功能以及抗氧化酶活性。结果表明，稀释液中添加400 μmol·L^-1 Met和200 μmol·L^-1AICAR均可显著提高解冻后精子活力、运动性能及精子结构完整性（P<0.05），其中400 μmol·L^-1 Met组精子总活力达43.20%，顶体完整率为91%，质膜完整率为46%。与对照组相比，Met组和AICAR组解冻后精子中AMPK磷酸化水平显著升高（P<0.05）；顶体酶活性显著提高（P<0.05）；丙酮酸水平显著下降（P<0.05），乳酸脱氢酶活性、乳酸以及ATP含量均显著升高（P<0.05）；与对照组相比，Met和AICAR组稀释液更有利于维持线粒体膜电位（P<0.05），提高ATP酶（P<0.05）以及抗氧化酶的活性（P<0.05）。添加适当浓度的AMPK激活剂可以提高绵羊精液冷冻保存的效果。  相似文献   

早胜牛的采精及细管冻精制作技术要点     

梁永虎 《现代农业科技》2023,(22)

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。  相似文献   

Recovery of sperms bearing X chromosomes with different concentrations of magnetic nanoparticles in ram     

Maryam Moradi  Hadi Hajarian  Hamed Karamishabankareh  Leila Soltani  Bijan Soleymani 《Reproduction in domestic animals》2021,56(2):263-269

Pre-conceptual sex selection is still a highly debatable process whereby X and Y chromosome bearing spermatozoa are isolated before oocyte fertilization. Recently, magnetic nanoparticles (MNP) have been used to determine X and Y chromosomes bearing spermatozoa as a result of searching for a cheap, highly efficient method using non-toxic materials. This study aimed to recover the sperm bearing X chromosomes in ram with different concentrations of MNP and then evaluate the success of this method using polymerase chain reaction (PCR). Ram sperms were divided into four groups, treated with 0 (control), 50, 100 and 200 μg/ml MNP, respectively. MNP was used to restore sperm cells bearing X chromosomes. Upon recovery, the PCR was performed to identify the X and Y sperms, Methyl ThiazoleTetrazolium (MTT), to assess MNP toxicity and sperm viability and acridine orange (AO) to evaluate sperm DNA integrity. The results of PCR revealed that the treatment of spermatozoa- bearing X chromosomes with 50 μg/ml MNP had the highest effects on the recovery of X sperm rather than the other concentrations of MNP. However, the concentrations of MNP did not have any toxic effects on spermatozoa, sperm viability and, DNA integrity, but the high concentration of MNP (200 μg/ml) significantly reduced DNA integrity. According to MTT and AO results, the concentrations of MNP used in this study had no toxic effects on spermatozoa and did not reduce the sperm viability and DNA integrity, except that 200 μg/ml MNP significantly reduced DNA integrity.  相似文献   

Production of monoclonal antibody against recombinant bovine sex-determining region Y (SRY) and their preferential binding to Y chromosome-bearing sperm     

Bijan Soleymani  Kamran Mansouri  Mohsen Rastegari-Pouyani  Shahram Parvaneh  Fatemeh Khademi  Mehdi Sharifi Tabar  Ali Mostafaie 《Reproduction in domestic animals》2021,56(2):270-277

Separation of X and Y chromosome-bearing sperm is an appropriate method for the selection of desired sex of offspring to increase the profit in livestock industries. The purpose of this study was the production of a monoclonal antibody against recombinant bovine sex-determining region Y protein for separation Y sperm. The hybridoma cells from splenocytes of immunized female's balb/C mice and Sp2/0 cells were made. The binding affinity of our monoclonal antibody (mAbSRY2) was compared with mouse monoclonal SRY-15. The Western blot method indicated that mAbSRY2 successfully detected the rbSRY protein. The specificity and sensitivity of mAbSRY2 is comparable to SRY-15 commercially ones. The SRY gene in 100% of bull semen contains the Y chromosome that had the strongest binding affinity to mAbSRY2 was synthesized. In other words, the binding affinity of semen contains the X sperms near the negative control. In general, this immunological method can help to separate X from Y sperms. However, the mAbSRY2 is bind to Y-bearing sexed sperm, but in the future; the sexed sperms need to apply in farms.  相似文献   

Variation among stallions in sperm quality after single layer centrifugation     

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   

Adjusted method of penis fixation during boar semi-automatic semen collection aiming to reduce bacterial contamination     

Aline Fernanda Lopes Paschoal  Ana Paula Gonçalves Mellagi  Cristina Vicenti Ferrari  Karine Ludwig Takeuti  Gabriela da Silva Oliveira  Mari Lourdes Bernardi  Rafael da Rosa Ulguim  Fernando Pandolfo Bortolozzo 《Reproduction in domestic animals》2021,56(6):897-904

Semen collection has an essential role in the initial bacterial load in boar ejaculates and extended semen. The study aimed to explore the efficacy of an adjusted penis fixation in a semi-automatic collection system on reducing bacterial contamination of ejaculates in two-boar studs with different scenarios. Historically, stud A had low levels of bacterial load in raw semen, while stud B had a high level of contamination. A total of 56 mature boars had their semen collected using two methods of penis fixation: (a) Traditional: The penis was fixed directly with the artificial cervix and transferred to the adjustable clamp; (b) Adjusted: The fixation was performed with one gloved-hand, and after exteriorization, the penis was gripped using the artificial cervix with the other gloved-hand and transferred to the adjustable clamp. The bacterial load (p = .0045) and the occurrence of ejaculates >231 CFU/ml (p = .0101) were reduced in the Adjusted compared to the Traditional method. Bacterial load was reduced when using the Adjusted method in stud B (p = .0011), which showed a greater occurrence of critical factors for bacterial contamination (p ≤ .0034). The Adjusted method reduced the occurrence of ejaculates >231 CFU/ml when the preputial ostium was dirty (p = .016) and the duration of semen collection was >7 min (p = .022) compared to the Traditional method. In conclusion, the Adjusted penis fixation was efficient in reducing bacterial load of ejaculates, mainly in boar stud B, which had high contamination challenges.  相似文献   

Study of microRNA Expression Profile in Different Regions of Ram Epididymis     

Cuiling Wu  Chunxin Wang  Bo Zhai  Yunhui Zhao  Zhuo Zhao  Zhiyu Yuan  Mingxin Zhang  Kechuan Tian  Xuefeng Fu 《Reproduction in domestic animals》2021,56(9):1209-1219

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Use of dimethylformamide to cryopreserve alpaca semen previously incubated with collagenase     

Nils H. Flores Huarco  Susana M. Giuliano  Fernanda G. Fumuso  Marcelo H. Miragaya  Hernán C. Cucho Dolmos  María I. Carretero 《Reproduction in domestic animals》2021,56(11):1387-1397

The objective of this study was to evaluate the effect of collagenase and two final dimethylformamide (DMF) concentrations (4% and 7%) on alpaca frozen-thawed sperm quality. A total of 25 ejaculates from 5 alpaca were obtained using electroejaculation. Each individual ejaculate was evaluated and then diluted 4:1 in a solution of 1 mg/ml collagenase in HEPES-TALP medium and incubated for 4 min at 37°C. Subsequently, samples were diluted in TRIS-fructose-citric acid-egg yolk and cooled to 5°C. Then, each sample was divided in two aliquots and DMF at final concentration of 4% or 7% was added, equilibrated for 1 hr at 5°C and frozen over liquid nitrogen vapours. A Kruskal–Wallis test was used to evaluate the sperm morphometry, and Completely Random Block designs were used to analyse sperm motility, viability, membrane function and acrosome status. After collagenase incubation, none of the samples showed thread formation, and sperm parameters were preserved. Non-progressive motile sperm were higher (p < .05) in equilibrated samples (4% DMF: 31.8 ± 8.3% and 7% DMF: 36.3 ± 11.8%) compared to raw (10.1 ± 4.3%) and frozen-thawed semen (4% DMF: 9.7 ± 1.8% and 7% DMF: 7.5 ± 3.2%). Sperm membrane function, membrane integrity and intact acrosomes were higher (p < .05) in raw semen (40.1 ± 12.2%, 94.6 ± 3.2% and 91.3 ± 8.1%) compared to frozen-thawed samples (4% DMF: 19.8 ± 4.7%, 53.2 ± 2.7%, 65.7 ± 8.7% and 7% DMF: 20.4 ± 4.5%, 54.1 ± 1.4%, 64.6 ± 9.1%). Length of the sperm head was lower in frozen-thawed samples, being statistically different with 4% DMF compared to pre-freezing samples. The ratio between acrosome and head areas was greater (p < .05) in frozen-thawed samples. Incubation of raw alpaca semen with collagenase decreased the thread formation without affecting sperm quality. Frozen of collagenase treated alpaca semen with 4% or 7% DMF did not preserve the sperm parameters in thawed samples.  相似文献