滴度及注射方式对2型腺相关病毒载体表达效果的影响

由于腺相关病毒载体(adeno-associated viral vector,AAV)本身的侵染不具有神经元特异性,其在神经系统相关研究中会存在侵染范围不符合试验要求的情况,因此本试验拟研究不同滴度和注射方式对病毒侵染范围的影响。结果显示,在注射较高滴度的AAV2-CMV-EGFP(1.3×10¹¹mg/L)3周后,其侵染范围可由延髓中缝苍白核(raphe pallidus nucleus,RPa)区域延伸至下丘脑室旁核(paraventricular nucleus,PVN)区域;较低滴度(1.3×10⁸mg/L)结合压力注射的方式可以在一定程度上限制AAV的侵染范围,并且载体携带的EGFP荧光基团在637 nm波段不具有明显的假阳性信号。     

Control of insect pests using trunk injection in a newly established apple orchard

We evaluated the potential for using preplant trunk injections of emamectin benzoate in nonbearing apple trees. Trees were evaluated for pest injury and emamectin residues throughout the planting season and into the following year. Injections into the trunk best delivered emamectin benzoate to the canopy compared with injections into the taproot, and the higher rate reduced insect pests more than the lower rate. In the following year, differences in insect control between trunk and root injections were less pronounced, but the higher rate of emamectin benzoate persisted longer and better reduced pests relative to the other treatments.     

Embryo-mediated genome editing for accelerated genetic improvement of livestock

Selecting beneficial DNA variants is the main goal of animal breeding. However, this process is inherently inefficient because each animal only carries a fraction of all desirable variants. Genome editing technology with its ability to directly introduce beneficial sequence variants offers new opportunities to modernize animal breeding by overcoming this biological limitation and accelerating genetic gains. To realize rapid genetic gain, precise edits need to be introduced into genomically-selected embryos, which minimizes the genetic lag. However, embryo-mediated precision editing by homology-directed repair (HDR) mechanisms is currently an inefficient process that often produces mosaic embryos and greatly limits the numbers of available edited embryos. This review provides a summary of genome editing in bovine embryos and proposes an embryo-mediated accelerated breeding scheme that overcomes the present efficiency limitations of HDR editing in bovine embryos. It integrates embryo-based genomic selection with precise multi-editing and uses embryonic cloning with elite edited blastomeres or embryonic pluripotent stem cells to resolve mosaicism, enable multiplex editing and multiply rare elite genotypes. Such a breeding strategy would enable a more targeted, accelerated approach for livestock improvement that allows stacking of beneficial variants, even including novel traits from outside the breeding population, in the most recent elite genetic background, essentially within a single generation.     

Management targets for continuously stocked mixed oat × annual ryegrass pasture in a no-till integrated crop–livestock system

Feeding livestock with cover crops can improve the efficiency and sustainability of integrated crop–livestock systems under no-till. However, no-till systems are based on permanent soil cover by organic material, so grazing livestock can compete for soil cover. Hence, managing stocking rates during the grazing period of the cultivated forage species is a key factor to assure enough herbage mass for maintaining long-term sustainable no-till systems. In this context, the objective of this study was to determine sward management targets for a continuously stocked mixed oat (Avena strigosa) × annual ryegrass (Lolium multiflorum) pasture in rotation with soybean in a no-till integrated crop–livestock system to determine the optimum balance between animal production and herbage mass for soil cover. The effects of sward height management on animal performance and herbage mass covers were evaluated. Treatments corresponded to four sward heights: 10, 20, 30 and 40 cm, maintained throughout the experimental period through continuous stocking and variable stocking rate, plus a no-grazing control area. Treatments were arranged in a randomized complete block design with three replications. Herbage mass and animal performance increased linearly with sward height, but weight gain per hectare decreased. Grazing efficiency fitted to a quadratic regression and conversion efficiency a logarithmic model. Equilibrium between grazing and conversion efficiencies was reached on swards managed at 20 cm, indicating that this sward height provided enough herbage mass to allow both animal performance and no-till crop demand for soil cover.     

Italian ryegrass establishment by self-seeding in integrated crop-livestock systems: Effects of grazing management and crop rotation strategies

We evaluated the re-establishment of an Italian ryegrass pasture by self-seeding on a no-till integrated crop-livestock systems (ICLS) in the southern region of Brazil. This work is part of a long-term experimental protocol initiated in 2003. We tested the effects of various management practices, such as summer crop systems (soybean vs. maize–soybean rotation), stocking methods (continuous vs. rotational) and grazing intensities (low vs. moderate), on Italian ryegrass pasture establishment. In addition, we tested resilience of the system by testing pasture's ability to re-establish following a year without seed head production. The experiment consisted in the rotation, on the same area, of Italian ryegrass pasture grazed by sheep during the winter and up to the end of the grass production cycle, and soybean or soybean-maize grain crops rotation cultivated during the summer. The pasture established itself by self-seeding since 2005. Data were collected in 2011 and 2012 stocking season. The soybean summer crop, continuous stocking and low grazing intensity, all positively affected the production of reproductive tillers in 2011. Grazing intensity in 2011 strongly influenced early vegetative tiller densities (before crop harvest) in 2012. However, none of the grazing intensity or the stocking method treatments affected herbage mass at the end of pasture establishment in 2011 or 2012. On the other hand, the soybean summer crop positively affected pasture establishment, both in term of tiller densities and herbage mass at the end of pasture establishment. The removal of all seed heads in 2011 (preventing seed production) resulted in the total failure of pasture establishment in 2012. Overall, Italian ryegrass establishment by self-seeding relies on the annual replacement of the soil seed bank. This experiment demonstrated that under various stocking methods, moderate grazing intensity and maize or soybean summer crop, Italian ryegrass pasture establishment by self-seeding remains successful even when the stocking periods extended up to the end of the grass production cycle. Self-seeding with moderate grazing intensity ensures successful pasture establishment, reduces labour and costs and allows to increase the stocking period and so animal live weight gain over the grazing season.     

单壁碳纳米管载嗜水气单胞菌外膜蛋白亚单位疫苗的研制及免疫效果评价

为了筛选细菌的疫苗候选蛋白,本实验根据课题组前期的研究,挑选了2个嗜水气单胞菌外膜蛋白(A0KLQ6和A0KH89),以A0KHR9(OmpAII)为对照,通过比较外膜蛋白与碳纳米管载外膜蛋白通过腹腔注射与浸泡免疫模式,比较和评价斑马鱼的免疫反应与免疫保护效果。结果显示,腹腔注射组和浸泡组与普通蛋白组对比,碳纳米管载蛋白能够显著提高斑马鱼免疫相关基因表达量。腹腔注射组在5μg剂量下免疫A0KHR9、A0KLQ6和A0KH89蛋白分别获得61.60%、 65.39%和84.96%相对免疫保护率(relative immune protection rate,RPS),在5μg剂量下免疫碳纳米管载蛋白SWCNTs-A0KHR9、SWCNTsA0KLQ6和SWCNTs-A0KH89分别获得68.10%、77.50%和90.43%的RPSs。浸泡组在40 mg/L剂量下免疫A0KHR9、A0KLQ6和A0KH89蛋白分别获得30.80%、25.85%和37.80%的RPSs,在40 mg/L最高剂量下免疫碳纳米管载蛋白疫苗SWCNTs-A0KHR9、SWCNTsA0KLQ6和SWCNTs-A0KH89分别获得63.60%、73.74%和68.49%的RPSs。研究表明,铁离子限制下嗜水气单胞菌外膜蛋白A0KLQ6和A0KH89等能够作为抗嗜水气单胞菌的亚单位疫苗候选蛋白,而单壁碳纳米管载蛋白通过注射免疫与浸泡免疫均能显著提升免疫疗效,是一种疫苗的适宜纳米载体。以上研究为寻找高效的嗜水气单胞菌亚单位疫苗候选蛋白及佐剂提供了理论依据。     

注射多巴胺对中华绒螯蟹打斗行为的影响

选同一家系规格相似、健康有活力的中华绒螯蟹新品种“光合1号”1龄雄蟹100只,分为4组,放入直径20 cm、高40 cm PE管(底部均匀钻12个直径为5 mm的孔洞)中,PE管置于120 L水槽中。用2个不透明烧杯将2只中华绒螯蟹分别倒扣于试验容器中,置于PE管两侧1 h。在中华绒螯蟹第三步足的动脉膜中分别注射20μL生理盐水(对照组),10^-8 mol、10-7 mol和10-6 mol的多巴胺液,利用闭路摄像头记录4 h内2只中华绒螯蟹打斗时长、次数、打斗分数及胜负关系等指标。打斗结束后,迅速取出中华绒螯蟹血淋巴,用ELISA酶联免疫法测定多巴胺含量。试验结果显示,对照组中华绒鳌蟹打斗时间最长,与各注射组差异显著(P<0.05);注射组中,随注射量的增加,打斗时长下降。对照组中华绒螯蟹打斗强度低于10^-8 mol组,高于其他注射组,与各注射组间差异显著(P<0.05)。注射组中,随注射多巴胺量升高,打斗强度下降,10^-8 mol组打斗强度最高,各组间差异显著(P<0.05)。对照组打斗次数与10-7 mol组差异不显著(P>0.05),与其余各组差异显著(P<0.05)。注射组中,随注射量的增加,打斗次数下降,10^-8 mol组与其余各组差异显著(P<0.05),10-7 mol组与10-6 mol组差异不显著(P>0.05)。打斗结束后,胜利方体内多巴胺含量与正常水平差异不显著(P>0.05),但显著高于失败方(P<0.05)。试验结果表明,注射多巴胺极显著影响中华绒螯蟹打斗行为(P<0.01),随多巴胺注射量的升高,中华绒螯蟹打斗行为整体下降。     

硫酸卡那霉素注射液含量测定的不确定度分析

为研究微生物检定法(管碟法)测定硫酸卡那霉素注射液生物效价试验中各因素对结果的影响,采用不确定度分析,将影响因素归纳为称量、稀释、加样、测定、平行试验,量化各不确定度分量,评估各因素的影响因子。结果表明,牛津杯加样体积对测定结果影响最大,提示试验中应尽可能精确控制加样体积。     

基于径向基神经网络的非寿险公司财务预警研究

选取我国保险行业中40家非寿险公司2006~2008年的混合非平衡面板数据作为研究样本，采用径向基神经网络模型从偿付能力、盈利能力、成长能力三个角度对非寿险公司财务预警进行研究。实证表明，此模型可以为管理层的财务预警提供更有有效的信息，并且在我国保险业具有样本量小，有效数据少的情况下，有着良好的运用前景。