Variation among stallions in sperm quality after single layer centrifugation

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.     

Sperm subpopulations influence the pregnancy rates in cattle

This study aimed to study the characteristics and subpopulations of spermatozoa from bulls with low and high reproductive performance based on pregnancy rates. Based on historical records of pregnancy rate from four farms, 24 bulls were selected. Two groups were established, with low pregnancy rates (n = 12; LOW), including bulls that presented pregnancy rates <52.27% (33.33% to 51.81%); and a group with high pregnancy rates (n = 12; HIGH), with pregnancy rates >52.27% (52.27% to 69.64%), after fixed-time artificial insemination (FTAI). The thawed sperm straws were analysed to sperm kinetics, morphology, plasma membrane integrity and sperm subpopulations. The LOW group exhibited a higher proportion of static cells (p < .05). In contrast, the HIGH group showed greater percentages for membrane integrity and total and progressive motility, and cells with fast and medium velocity (p < .05). In the cluster procedures, four sperm subpopulations were established. The low-fertility bulls presented the highest percentage of subpopulation 2 (41.46%), characterized by slow and progressive spermatozoa. The high-fertility bulls exhibited the highest percentage of subpopulation 3 (37.17%), characterized by fast and nonlinear spermatozoa. Results from this study indicated that bulls with greater percentages of fast and nonlinear spermatozoa seem to have greater fertilization capacity and the subpopulations analysis can be considered a tool to identify ejaculates with high fertility.     

Use of dimethylformamide to cryopreserve alpaca semen previously incubated with collagenase

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.     

自然保护区和国有林区野猪在贵州省道真县境内死亡的流行病学调查

2020年2月20日,贵州省道真县生态护林员在其境内的国家级大沙河自然保护区和国有林区发现多具野猪尸体。为弄清野猪死亡原因,防止可能出现的疫情在当地扩散,采用描述性流行病学方法,对本次野猪死亡事件展开调查。截至2020年3月4日,在该县3个镇8个村(社区)10个村民组,共排查发现23具野猪尸体,以体质量偏小的低幼龄野猪居多,约占70%;发现地点分布具有明显的聚集性和方向性;因大部分尸体高度腐败,仅从采集的1份病料中,检出猪圆环病毒核酸阳性,未检出猪瘟、口蹄疫、猪繁殖与呼吸综合征、非洲猪瘟等疫病病原;经排查,未发现野猪尸体附近的村(社区)生猪养殖户饲养的生猪发生异常发病死亡现象。经伽马分布估算,该县平均每天可能出现的死亡野猪数为0.44头(95%CI,0.28~0.64)。综上分析推测,野猪死亡既有经境内公路来往的生猪运输车辆及废弃物传入疫病导致的可能性,也不排除中毒或天气变化等其他因素的可能。由此建议加强公路沿线一带的林区排查,加大跨省公路车辆消毒力度,降低野猪死亡事件带来的潜在风险。     

Cryopreservation of ram semen: Manual versus programmable freezing and different lengths of equilibration

The aim of our study was to examine effects of the length of semen equilibration as well as two freezing techniques on ram sperm post-thaw quality. The ejaculates of Wallachian sheep rams (n = 12) were collected by an electro-ejaculation, equilibrated in a Triladyl® (0, 2, 4, 6, and 8 h) containing glycerol and egg yolk and frozen by programmable freezing (PF) or manual freezing (MF). After thawing, sperm samples were subjected to the motility (computer-assisted sperm analysis [CASA]), viability (SYBR-14/PI), and fertilizing ability (FA) (in vitro penetration/fertilization test on bovine oocytes) assays. It was found that the equilibration of 6 h (E-6) ensured higher post-thaw sperm motility and progressive movement compared with other lengths tested, irrespective of a freezing technique. The E-6 sperm viability did not differ between PF and MF but was lower (P < 0.05) than control. Sperm FA (E-6) was similar in PF (60.44%) and MF (62%) but slightly lower than in fresh (72.8%). Our data demonstrate that the use of MF was comparable with PF, which can be applied in the field conditions without need in a piece of cost-expensive equipment, which can greatly benefit the gene bank of animal genetic resources.     

Effect of Se and VE Supplement on Semen Quality,Antioxidant Enzyme Activities and Heat Shock Protein Expression of Goat in High Temperature Season

This experiment was conducted to study the effect of selenium and vitamin E supplement on semen quality, antioxidant enzyme activities and heat shock protein expression of goat in Hainan high temperature season.16 adult Hainan Black goat with good health and approximate weight were randomly divided into 4 groups, fed with basal diet(control group), basal diet+0.5 mg/kg Se(Se group), basal diet+100 mg/kg VE(VE group), and basal diet+0.5 mg/kg Se+100 mg/kg VE(Se+VE group), respectively.The experimental period was 93 d.Semen samples were collected in the last week of the experiment on two consecutive days.The semen quality, antioxidant enzyme activities and heat shock protein expression were analyzed.The results showed that compared with control group, the ejaculate volume was not significantly affected by Se or VE supplement(P>0.05).Sperm density and sperm motility were increased significantly by Se and VE supplement(P<0.05), and the abnormal rate was decreased extremely significantly(P<0.01).The goats fed with Se and VE also had higher activities of GSH-Px(P<0.01), SOD(P<0.05), CAT(P<0.05) and T-AOC(P<0.01), and lower MDA concentration in seminal plasma(P<0.05).The relative expression levels of HSP70 and HSP90 mRNA in supplement groups were decreased extremely significantly(P<0.01).However, there were some certain differences between the Se and VE supplement groups on semen quality and heat shock protein expression.In conclusion, the supplementation of Se and VE could help to improve goat semen quality by increasing the sperm density, sperm motility, the antioxidant enzyme activities, and decreasing the abnormal rate in hot season of Hainan.Finally, Se and VE supplement had good effects on relieving the environment heat stress.     

呼玛县林业局野生蓝莓人工抚育技术探讨

文章详细介绍了呼玛县林业局野生蓝莓分布、生物学特性以及提高野生蓝莓产量和质量的抚育技术。     

稀释液中添加海藻糖对猪冷冻精液质量参数和活性氧的影响

本试验旨在探讨海藻糖对猪精子冷冻效果以及精子内活性氧(ROS)的含量影响。试验分对照组和4个海藻糖处理组(0.025,0.05,0.1和0.2 mol/L)。精子冷却后,添加0.1 mol/L海藻糖处理组精子活力显著(P<0.05)高于对照组,而精子内和稀释液中各组间ROS发生量都没有显著的差异。精子冷冻解冻后,添加0.05 mol/L海藻糖处理组的精子活力显著(P<0.05)高于对照组;添加0.05 mol/L和0.1 mol/L海藻糖处理组的精子生存率显著(P<0.05)高于对照组;添加0.1 mol/L海藻糖处理组顶体完整的精子百分比显著(P<0.05)高于对照组;4个海藻糖处理组膨胀精子百分比都显著(P<0.05)高于对照组;添加0.025 mol/L海藻糖处理组ROS水平显著(P<0.05)低于对照组。结果表明:海藻糖对精子冷冻保存是有益的,且能抑制精子内ROS的发生,但它的作用机制有待于进一步的研究。     

日粮维生素E水平对山羊精液品质的影响

采用4×4拉丁方设计,给金堂黑山羊饲喂维生素E浓度不同(常量:30 mg/kg维生素E,10,50,100倍常量的添加量)而其他营养成分均一致的4种日粮,探索日粮中添加维生素E对山羊精液品质的影响。结果表明:日粮中添加维生素E有助于提高山羊采精量、精子密度和鲜精活率,改善其精液品质(P<0.05);随着日粮中维生素E浓度的增加,山羊采精量、精子密度和鲜精活率也随之改善,添加量为常量的50倍(1500 mg/kg)时,精子密度和鲜精活率最佳(P<0.01)。当添加量超过常量50倍时(100),精子密度和鲜精活率有所降低,但与添加量为50倍的试验组差异不显著(P>0.05)。