恒定水头供水瓶的性能分析

恒定水头供水瓶的弹性构件弹簧竖立在敞口容器和支座之间,该装置通过水体自重与弹簧弹力相互作用使敞口容器内的上液面保持绝对高程不变,使水头保持稳定。在实际应用中,可以在一定限度内自由增减敞口容器内水量,避免了马氏瓶在实验前需要放水以维持水头稳定与实验过程中不能对瓶内水量做任何控制以及该仪器不适于高温环境中使用的弊端,且该仪器可以缩小体积,方便拆装与携带。经计算,该装置系统误差在4%以内。研究提出的恒定水头供水瓶,原理简单,适应性强,可在各种入渗、蒸发实验中用于控制水头高度的恒定,具有广泛的应用前景。     

A simplified roller bottle platform for the production of a new generation VLPs rabies vaccine for veterinary applications

Rabies is a neglected disease with an estimated annual mortality of 55,000 human deaths, affecting mainly low-income countries. Over 95% of these cases result from virus transmission through the bite of infected dogs and for this reason there is a real need for a cheap and effective rabies veterinary vaccine to be used in mass vaccination campaigns. In this work, we describe the establishment of a simple platform for the production of a virus-like particles based rabies vaccine using mammalian cells and roller bottles as culture system. Adherent cells were cultured during more than 15 days and VLPs were continuously produced and secreted to the culture supernatant. Immunogenicity and protective efficacy of VLPs were tested through rabies virus neutralizing antibody test and NIH potency test. These viral particles induced high titer of long lasting neutralizing antibodies and protected mice against active virus challenge. Therefore, this development represents a promising platform for the production of a new generation and virus-free rabies vaccine candidate for veterinary applications.     

侵染广东连州葫芦的黄瓜绿斑驳花叶病毒的分子特征 及致病性分析

【目的】黄瓜绿斑驳花叶病毒(Cucumber green mottle mosaic virus,CGMMV)是侵染瓜类作物的主要病毒之一,对瓜类产业造成巨大的危害。本研究旨在探明侵染广东省连州市葫芦的CGMMV分离物(CGMMV-GDLZ)分子特征及其在系统进化中的地位,并测定其对黄瓜、葫芦和西瓜的致病性,为CGMMV的防控提供理论依据。【方法】从广东省连州市葫芦种植基地采集2个疑似CGMMV侵染的病样以及1个无症状样品,提取总RNA,根据CGMMV参考序列（GenBank登录号：KX883801）设计引物进行RT-PCR检测,引物序列为F：CCACGAGTTGTTTCCTAATGCTG/R：TTTGCTAGGCGTGATCGGATTGT,退火温度53℃,扩增长度890 bp。将CGMMV全长序列分为前后两段,前半段1—3 511 nt,扩增引物序列为F：AAGTTCATTTCATTTGGAGAGGGTTTTAATTTTTATAA TTAAACAAA/R：AGTTCTGCATTAATTGCTATTTGGTAGGCACAGTGGTAG;后半段3 301—6 423 nt,扩增引物序列为F：GTGCGTGCTACCCCGACTCCAATAGGTTTGATTGCCCGTG/R：GGTGGAGATGCCATGCCGACCCTGGGCCCCTACCCGGGGAAAGG。将前后两段PCR产物通过同源重组的方法克隆到pCB301双元载体上,测序得到CGMMV-GDLZ分离物全长序列。利用CGMMV-GDLZ分离物全长序列在NCBI中进行Blast分析,然后通过MEGA7软件对CGMMV-GDLZ以及其他已经报道的CGMMV分离物进行系统进化树分析。将构建好的pCB301-CGMMV侵染性克隆注射接种本生烟验证其侵染性,然后再注射接种黄瓜、葫芦和西瓜的子叶,测定CGMMV-GDLZ分离物的致病性。【结果】RT-PCR结果证实,广东省连州市葫芦病样感染了CGMMV。CGMMV-GDLZ分离物全长序列为6 423 nt,编码4个蛋白,分别为129K复制酶（61—3 495 nt）、186K复制相关蛋白（61—5 007 nt）、运动蛋白MP（4 994—5 788 nt）和外壳蛋白CP（5 763—6 248 nt）。CGMMV-GDLZ核苷酸序列与CGMMV-eWT分离物（GenBank登录号：KY753928）同源性最高,为99.97%。系统进化树分析结果显示,CGMMV-GDLZ分离物与日本、韩国等东亚CGMMV分离物同属Group 1,在遗传距离上与山东、浙江和河南的CGMMV分离物最接近。pCB301-CGMMV侵染性克隆可以系统侵染本生烟,造成本生烟上部叶片出现皱缩、斑驳、凸起等症状,RT-PCR和Western blot进一步确认了侵染性克隆的侵染性。注射接种CGMMV-GDLZ分离物后15 dpi,葫芦和西瓜即可产生斑驳、花叶、突起、生长迟缓等症状,24 dpi时症状更明显。而15 dpi时,CGMMV-GDLZ分离物在黄瓜上的症状不明显,与未接种对照植株几乎没有区别;将植株从控温接种室移入网室中,30 dpi时,黄瓜植株上部叶片开始出现斑驳和花叶,40 dpi时,症状已经非常明显。RT-PCR和Western blot检测进一步确认了上述结果。【结论】侵染广东省连州市葫芦的CGMMV-GDLZ分离物与山东、浙江和河南的CGMMV分离物很可能具有相同的传染源;CGMMV-GDLZ分离物可以侵染本生烟、黄瓜、葫芦和西瓜等作物,但对这些作物的致病性存在差异。     

应用正交试验优化猪细小病毒N株在IBRS-2细胞转瓶培养

利用正交试验对猪细小病毒(PPV)N株在IBRS-2细胞转瓶培养中的培养条件进行优化,为PPV N株弱毒疫苗的大量培养工艺提供参考.设细胞培养液pH值(7.0、7.2和7.4)、接毒时间(0、24和48 h)、接毒剂量(0.10％、1.00％和10.00％)、收毒时间(48、72和96 h)等因子和水平.结果表明,PPV N株转瓶培养的最佳培养条件组合为细胞培养液pH值7.2,采用同步接毒,接毒剂量为0.10％,收毒时间为接毒后96 h.其中收毒时间是影响PPV N株毒价的最主要因素.     

棉花黄萎病抗性鉴定新方法——无底塑钵菌液浇根法

通过一系列的试验 ,笔者发展了一种快速鉴定棉花品种抗黄萎病性能的新方法—无底塑钵菌液浇根法 ,此法比传统的纸钵撕底醺根法更简便 ,发病较轻 ,能准确客观地反映品种的抗性     

一种新的厌氧污泥比活性试验测定法

本文介绍了一种以医用血清瓶作发酵瓶、用医用注射器来量测所产沼气的厌氧污泥比活性试验测定法。实践表明此法简单实用、快速准确。     

葡萄组培苗瓶外生根技术研究

采用不同种类和浓度的生长素诱导葡萄组培苗瓶外生根，并观察组培苗在不同处理基质、不同的温度、不同的生理年龄条件下生根的效果。通过优化技术参数，建立了葡萄组培苗瓶接生根的技术优化体系，结果表明，用浓度为0.3mg/L的IBA浸蘸处理的插条，在温度为27~29℃的环境条件下在基质珍珠岩上培养，瓶苗茎段的生根率可达85.5 %，移栽后苗木的成活率达98 %，相对于其他方法有了较大的提高。该技术的建立有助于实现葡萄工厂化育苗技术的商业化扩大。     

吊瓶精准输液技术对芒果产量的影响

采用吊瓶精准输液技术，在芒果树开花到成熟期按树体大小定量向树杆木质部注射营养液，以确定对芒果产量的增产效应，结果表明：芒果果数可增产23.85%，单果重可增产22.77%，总产量可增产52.06%，其产投比为3.38:1，是对照产投比（2.29:1）的1.48倍。通过定量分析和计算机模拟的方法推导出芒果精确施肥用药量的数学模型为?=5.15383(D)3.09674(g)，对吊瓶输液技术导致芒果抗旱丰产的原因进行分析及讨论。     

组培洗瓶机的水力洗涤工艺研究

采用模型试验的方法对组培洗瓶机的水力洗涤工艺进行了研究，论证了水力洗涤方式完全可满足组培洗瓶的要求，确定了水力洗涤各参数对洗涤效果的影响程度，并获得了水力洗涤主要参数的较佳范围，为设计组培洗瓶机提供了理论依据。     

The effect of exogenous purine supply on the endogenous excretion of purine derivatives in the urine of growing lambs

An experiment was made to determine the absorption of purine metabolites in dietary nucleic acids through the digestive tract, and also to determine the utilization of nucleic acids absorbed in the body, using growing lambs. Two pairs of 120‐ and 180‐day‐old twin female lambs with a bodyweight of 18.2–19.0 kg were kept in metabolism crates and fed on purine‐free milk replacement (MR) with supplements of exogenous purine (purine base or purine nucleoside) at a level of 0.2 mmol/BW^0.75/d for 5 consecutive days, and thereafter they were maintained in the crates for 4 days. The daily amount of exogenous purine supply was calculated based on the urinary excreted purine derivatives (PD) in lambs fed on milk replacement alone. A urine sample was collected daily for 9 consecutive days, and the urinary excretion of PD was determined daily. Urinary PD excretion opened to increase within 24 h after the dose of purine bases, and the level was recovered on 3 days after ceasing the exogenous purine supply. The recovery of PD in the urine was about 70% of the purine supplement. When purine nucleosides were added to the feed, urinary PD excretion was initiated within 24 h after dosing, and the values were recovered after ending the purine nucleoside supply. The recovery rate of PD in the urine was only 30% of the supplemented purine. The plasma allantoin levels were almost similar after feeding purine bases and purine nucleosides, and the values were mostly in the range (40–60 µmol/L). These findings indicate that an exogenous purine can be directly incorporated into the body, and the purine as nucleoside is more effectively utilized for the synthesis of nucleic acids than as a purine base in the body of growing lambs.