鸡传染性支气管炎的多重RT-PCR检测及793／B血清型的鉴定

【目的】建立鸡传染性支气管炎病毒(IBV)快速、准确的检测方法。【方法】根据GenBank中已经发表的793/B型和多株IBV的N基因和S1基因,对比分析后以N基因的保守序列设计合成了1对引物,跨度为582bp;以793/B的S1基因设计合成了1对特异型引物,跨度为891 bp;建立了能检测出IBV并能够同时鉴定出793/B血清型的实验室一次性PCR诊断方法。【结果】多重RT-PCR检测表明,793/B血清型可出现582和891 bp 2条核酸片段,而其他血清型只出现582 bp 1条核酸片段,新城疫病毒、传染性喉气管炎、鹅副粘病毒则无任何条带出现。【结论】所建立的TR-PCR方法具有一定的特异性,可用于IBV的快速检测和793/B血清型的临床诊断。

鸡传染性支气管炎病毒变异株(793/B)核蛋白基因的克隆与序列分析

根据GenBank已经发表的传染性支气管炎病毒(IBV)N全基因组序列设计引物,对IBV793/B分离毒株N基因进行克隆与序列分析。结果表明,IBV793/B的N基因由1229bp组成,与GenBank已发表的11株IBV的N基因相比较,IBV793/B的N基因共有88处点突变,在第991位发生了一个核苷酸的缺失。N基因的核苷酸同源性为86.9%～91.4%,氨基酸同源性为75.8%～77.5%。表明IBV93/B的N基因存在着较大的变异性。

Screening for the Strain Highly Producing Antagonistic Substance from Bacillus subtilis B47 by UV Mutagenesis

[Objective] The aim of this study was to breed new strains which have higher inhibitory effects on the pathogens of watermelon fusarium wilt.[Method] The endophytic Bacillus subtilis B47 strain was obtained from tomato stems by UV mutagenesis for two consecutive times,then genetic stability as well as physiological and biochemical properties of mutant strains were studied.[Result] The antibacterial activity of all the three mutant strains F303,F304 and F305 was higher than that of B74 strain.After subculture of 10 successive generations,the antibacterial activity of all the three mutant strains for the pathogens of watermelon fusarium wilt decreased,but the antibacterial activity of F305 strain decreased the least,indicating its best genetic stability among the tested strains.The antibacterial circle diameter of F305 strain was 5 mm larger than that of wild strain B47 under the same condition.The mutant strain F305 was in logarithmic growth phase within 36 h and in stationary phase within 36-96 h,while its optimum growth temperature was 35 ℃.F305 strain could grow in sodium salt with the concentration of 1%-10%,but it grew best at the concentration of 1%.Physiological and biochemical responses of F305 strain were in accordance with those of wild strain B47.[Conclusion] This study lays the foundation for the factorial production of antagonistic substance by B47 strain and new methods of preventing from the pathogens watermelon fusarium wilt.

鸡传染性支气管炎病毒793/B RT-PCR检测方法的建立及应用

根据GenBank已发表的鸡传染性支气管炎病毒(Infectious bronchitis virus,IBV)793/BS1基因序列,设计1对引物,扩增891bp特异性核酸片段,建立了检测IBV793/B的RT-PCR方法。特异性试验结果表明,IBV793/B能扩增出891bp的核酸片段,而IBVM41、H120、H52毒株以及新城疫病毒(NDV)、禽流感病毒(AIV)、传染性法氏囊病病毒(IBDV)均无特异性条带出现。敏感性试验结果表明,该方法的最低检出量为10pg的模板。上述结果表明,本试验所建立的RT-PCR方法敏感性高、特异性强。利用建立的RT-PCR方法对从山东省分离的8株疑似鸡IBV793/B进行检测,结果7株为阳性。该方法的建立为IBV793/B的诊断及流行病学调查提供了可靠的方法。

Influence of strain rate on microstructure and formability of AZ31B magnesium alloy sheets

The effects of strain rate on microstructure and formability of AZ31B magnesium alloy sheets were investigated through uniaxial tensile tests and hemispherical punch tests with strain rates of 10^-4, 10^-3, 10^-2, 10^-1 s^-1 at 200℃. The results show that the volume fraction of dynamic recrystallization grains increases and the original grains are gradually replaced by recrystallization grains with the strain rate decreasing. A larger elongation and a smaller r-value are obtained at a lower strain rate, moreover the erichsen values become larger with the strain rate reducing, so the formability improves. This problem arises in part from the enhanced softening and the coordination of recrystallization grains during deformation.

Construction and expression of eukaryotic plasmids containing lamivudine-resistant or wild-type strains of Hepatitis B Virus genotype C

AIM:To construct eukaryotic expression plasmids of full-length Hepatitis B Virus(HBV) genotype C genome,which contain lamivudine-resistant mutants(YIDD,YVDD) or wild-type strain(YMDD) ,and to observe the expression of HBV DNA and antigens [hepatitis B surface antigen(HBsAg) and hepatitis B e antigen(HBeAg) ] of the recombinant plasmids in HepG2 cells. METHODS:Three HBV full-length genomes were amplified from the plasmids pMD18T-HBV/YIDD,pMD18T-HBV/YVDD and pMD18T-HBV/YMDD,using PCR. Three recombinant plasmids were generated by inserting each of the PCR products into the eukaryotic expression vector pcDNA3.1(+) ,between the EcoRI and HindⅢ sites. After being characterized by restriction endonuclease digestion,and DNA sequence analysis,the recombinant plasmids were transfected into HepG2 cells. At 48 and 72 h post-transfection,the levels of intracellular viral DNA replication were detected by real-time PCR,and the expression of HBsAg and HBeAg in the cell culture supernatant was determined by ELISA. RESULTS:Restriction endonuclease digestion and DNA sequence analysis confirmed that the threerecombinant plasmids were correctly constructed. After transfecting the plasmids into HepG2 cells,high levels of intracellular viral DNA replication were observed,and HBsAg and HBeAg were secreted into the cell culture supernatant. CONCLUSION:Eukaryotic expression plasmids pcDNA3.1(+) -HBV/YIDD,pcDNA3.1(+) -HBV/YVDD or pcDNA3.1(+) -HBV/YMDD,which contained HBV genotype C full-length genome,were successfully constructed. After transfection into HepG2 cells,the recombinant plasmids efficiently expressed HBV DNA,HBsAg and HBeAg. Our results provide an experimental basis for the further study of HBV lamivudine-resistant mutants.

Hydrogen Production with High Evolution Rate and High Yield by Immobilized Cells of Hydrogen-producing Bacteria Strain B49 in a Column Reactor

To improve the hydrogen evolution rate in continuous hydrogen production of a fermentative hydrogen-producing bacteria strain B49 (AF481148 in EMBL), 4% immobilized cells by polyvinyl alcohol-boric acid method, with the addition of a small amount of calcium alginate in a column reactor obtain hydrogen yield of 2.31 mol H2/mol glucose and hydrogen evolution rate of 1435.4 ml/L culture&middoth respectively at medium retention time of 2 h with a medium containing l0 g glucose/L. As the cell density in gel beads is increased to 8%, hydrogen yield and hydrogen evolution rate for l0 g glucose/L are 2.34 mol H2/mol glucose and 2912.4 ml/L culture &middot h respectively at medium retention time of 1 h, and for molasses wastewater COD of 7505.9 mg/L hydrogen production potential of 205.6 ml/g COD and hydrogen evolution rate of 2057.7 ml/L culture&middoth at hydraulic retention time of 0.75 h are observed. In the continuous culture pH value keeps around 3.9 by self regulation.

Mechanical characterization of Mg-B4C nanocomposite fabricated at different strain rates

Magnesium has wide application in industry.The main purpose of this investigation was to improve the properties of magnesium by reinforcing it using B4C nanoparticles.The reinforced nanocomposites were fabricated using a powder compaction technique for 0,1.5vol%,3vol%,5vol%,and 10vol% of B4C.Powder compaction was conducted using a split Hopkinson bar(SHB),drop hammer(DH),and Instron to reach different compaction loading rates.The compressive stress–strain curves of the samples were captured from quasi-static and dynamic tests carried out using an Instron and split Hopkinson pressure bar,respectively.Results revealed that,to achieve the highest improvement in ultimate strength,the contents of B4C were 1.5vol%,3vol%,and 3vol% for Instron,DH,and SHB,respectively.These results also indicated that the effect of compaction type on the quasi-static strength of the samples was not as significant,although its effect on the dynamic strength of the samples was remarkable.The improvement in ultimate strength obtained from the quasi-static stress–strain curves of the samples(compared to pure Mg)varied from 9.9% for DH to 24% for SHB.The dynamic strength of the samples was improved(with respect to pure Mg)by 73%,116%,and 141%for the specimens compacted by Instron,DH,and SHB,respectively.The improvement in strength was believed to be due to strengthening mechanisms,friction,adiabatic heating,and shock waves.

Partial Characteristics of Hydrogen Production by Fermentative Hydrogen-producing Bacterial Strain B49

To investigate the characteristics of hydrogen production by a novel fermentative hydrogen-producing bacterial strain B49 (AF481148 in EMBL), batch experiments are conducted under different conditions. Hydrogen production has a correlation with cell growth and the consumption of glucose and soluble protein. The optimum pH for cell growth is 4.5±0.15. At acidic pH 4.0±0.15, the bacteria has the maximum accumulated hydrogen volume of 2382 ml/L culture and the maximum hydrogen evolution rate of 339.9 ml/L culture·h with 1% glucose. The optimum temperature for cell growth and hydrogen production is 35℃. In addition, fermentative hydrogen-producing bacterial strain B49 can generate hydrogen from the decomposition of other organic substrates such as wheat, soybean, corn, and potato. Moreover, it can also produce hydrogen from molasses wastewater and brewage wastewater, and hydrogen yields are 137.9 ml H 2/g COD and 49.9 ml H 2/g COD, respectively.

Magnesium improves hydrogen production by a novel fermentative hydrogen-producing bacterial strain B49

Batch experiments were conducted to investigate the effects of magnesium on glucose metabolism, including growth and hydrogen-producing capacity of fermentative hydrogen-producing bacterial strain B49. These abilities were enhanced with an increase in magnesium concentration. At the end of fermentation from （10 g/L） glucose, for 10 mg/L MgCl2·6H2O the cell growth in terms of optical density （OD） at 600nm was 0.46, the ratio of ethanol amount （mg/L） to acetate amount （mg/L） was 1.1, and the accumulated hydrogen volume was 934.9 mL H2/L culture; for 200 mg/L of MgCl2·6H2O OD600 nm was increased to 1.34. The accumulated hydrogen volume was increased to 2 360.5 mL H2/L culture, the ratio of ethanol amount （mg/L） to acetate amount （mg/L） was increased to 1.3 and polysaccharide was decreased to 2.5 mg/L. Moreover, the magnesium solution addition to the medium at different fermentation times affected hydrogen-producing ability. However, the later the addition time was postponed, the less the effect was on hydrogen evolution. Further experiments confirmed the enhancement was dependent on magnesium ions and not on the other inorganic ions such as SO42- or Cl-, which constituted the magnesium salts.

基于声发射b值演化的脆性岩石裂纹起裂应力评价方法

岩石裂纹起裂应力的识别和研究对认识脆性岩石受载变形及破坏过程具有重要意义。系统总结了现有的裂纹起裂应力确定方法(裂纹体积应变法、侧向应变法、移动点法、声发射振铃计数法)及存在的问题,并以单轴压缩条件下岩石破裂过程中声发射幅值及b值分析为基础,提出了一种基于声发射b值演化的脆性岩石裂纹起裂应力评价方法。采用GAW2000单轴刚性试验机和声发射测试仪开展了一系列变辉长岩和花岗岩的单轴加载试验,根据力学及声发射测试结果,对所提方法及现有方法进行了评价。研究结果表明:声发射b值演化法具有明确的物理机制及意义,可以方便准确地确定起裂应力大小,测定应力结果与现有方法所得结果具有一致性。通过方差分析可知,所提方法与现有方法不存在统计差异,验证了该方法测定裂纹起裂应力的可靠性。2类岩样的起裂应力均与单轴抗压强度呈正线性相关,使用所提方法确定的变辉长岩起裂应力约为单轴抗压强度的0.51倍,花岗岩起裂应力约为单轴抗压强度的0.48倍。所提方法及研究成果对于丰富岩石裂纹起裂应力测试手段和探究岩石脆性破坏机制具有一定的价值。

B亚型禽偏肺病毒减毒株对商品肉鸡免疫效果的评价

旨在研究B亚型禽偏肺病毒减毒株(LN16-A株)对商品肉鸡的免疫效果。本研究从规模化养殖场选取3周龄的商品肉鸡,将B亚型禽偏肺病毒减毒株(LN16-A株)通过滴鼻方式免疫(200μL,病毒含量≥103 TCID50·只^(-1)),空白对照组以同样剂量的PBS滴鼻。免疫21 d后,翅下采集血液分离血清,利用ELISA方法及中和试验检测相应抗体;在6周龄时,通过滴鼻方式使用B亚型禽偏肺病毒强毒(LN16-V株)(200μL,5000 TCID50·只-1)进行攻毒,攻毒后连续观察7 d,并利用RT-qPCR检测各组鸡鼻腔拭子的病毒拷贝数,攻毒后第9天各组随机选取2只鸡进行剖杀,采集鼻甲骨、气管和肺组织,通过HE染色制作病理切片,观察组织病理学变化。抗体检测结果显示,在疫苗免疫后21 d,免疫组ELISA抗体和中和抗体阳性率均为100%,平均ELISA抗体效价为1860,平均中和抗体效价为7.44 log2;临床症状观察结果显示,对照组商品肉鸡出现混浊、黏稠、牵丝状的鼻液等明显临床症状,发病率为100%,而免疫组肉鸡无临床症状;RT-qPCR结果表明,免疫组鸡鼻腔拭子病毒基因组拷贝数相较于对照组降低了78%~96%;病理切片结果显示,对照组肉鸡鼻甲骨、气管以及肺均出现不同程度的病理损伤,免疫组肉鸡各组织器官未见明显病理变化。综上表明,B亚型禽偏肺病毒减毒株(LN16-A株)可以诱导商品肉鸡产生特异性抗体,能显著降低攻毒后的排毒水平,对商品肉鸡具有良好的保护效果。本研究结果将为肉鸡养殖场禽偏肺病毒病的防控提供重要理论指导和技术支撑。

Influencing Factors and Process on <i>in Situ</i>Degradation of Poly(<i>Butylene Succinate</i>) Film by Strain <i>Bionectria ochroleuca</i>BFM-X1 in Soil

This is the first report on the PBS film degraded by any Bionectria ochroleuca fungal strain. The fungal strain BFM-X1 was isolated from an air environment on a vegetable field and was capable of degrading poly(butylene succinate) (PBS). The taxonomic identity of the strain BFM-X1 was confirmed to be Bionectria ochroleuca (showing a 99% similarity to B. ochroleuca in a BLAST search) through an ITS rRNA analysis. The bio-degradation of the PBS film by strain BFM-X1 was studied. Approximately 97.9% of the PBS film was degraded after strain BFM-X1 was inoculated at 28?C for 14 days. The degradation efficiency of BFM-X1 against PBS film under different soil environmental conditions was characterized. The results indicated that 62.78% of the PBS film loss was recorded in a 30-d experimental run in a sterile soil environment indoors. On adding strain BFM-X1 to a soil sample, the PBS degradation rate accelerated approximately fivefold. Furthermore, both temperature and humidity influenced the in situ degradation of the PBS by strain BFM-X1, and temperature may be the major regulating factor. The degradation was particularly effective in the warm season, with 90% of weight loss occurring in July and August. Scanning electron microscope observations showed surface changes to the film during the degradation process, which suggested that strain BFM-X1preferentially degraded an amorphous part of the film from the surface. These results suggested that the strain B. ochroleuca BFM-X1 was a new resource for degrading PBS film and has high potential in the bioremediation of PBS-plastic-contaminated soil

猴痘病毒B.1谱系遗传分支、毒力基因及蛋白功能

2022年以来,猴痘疫情在全球暴发和流行。相较以往的猴痘病毒,2022年流行的猴痘毒株传播能力和宿主适应性等明显增强,猴痘B.1谱系毒株已成为全球猴痘疫情流行的主要毒株。为此,本文对猴痘病毒B.1谱系遗传分支、毒力基因及蛋白功能进行综述,并就部分基因产物的蛋白功能进行了注释,以期为猴痘疫情的科学防控提供参考。

马立克氏病病毒(MDV)814株B抗原基因的克隆及部分序列分析

提取感染鸡胚成纤维细胞ＭＤＶ－Ｉ弱毒株８１４病毒ＤＮＡ为模板，根据ＲＢＩＢ株ｇＢ基因５＇及３＇两端核苷酸序列设计引物，利用ＰＣＲ技术扩增了我国ＭＤＶ－Ｉ弱毒株８１４ｇＢ基因（１．９ｋｂ），将扩增片段平末端克隆到载体ｐＢｌｕｅｓｃｒｉｐｔＳＫ中ＥｃｏＲＶ位点，经ＢａｍＨＩ、ＨｉｎｄⅢ酶切鉴定得到不同插入方向的重组质粒。构建扩增片段的酶切图谱及部分序列分析证明与ＲＢＩＢ株ｇＢ基因无差异，显示了极高的同源性。本文进一步证明ｇＢ基因在ＭＤＶ不同毒株间是十分保守的。

酵母菌Y48对活性黑KN-B的脱色研究

从土壤中分离获得了一株对偶氮染料具有明显脱色效果的酵母菌Pseudozyma rugulosaY48,其对初始浓度为100 m.gL-1的活性黑KN-B的最大脱色率可达99%.采用酵母菌Y48对偶氮染料脱色的优化控制条件是:接种量应不低于3%(体积分数);最适pH值在6—9之间;氮源(NH4)2SO4的浓度不低于0.1%;碳源葡萄糖的浓度为1%.对其脱色机理的研究表明,Y48对活性黑KN-B的脱色是由吸附和降解两个过程完成的,而对活性黑KN-B分子的去除则主要依靠生物降解.此研究表明,酵母菌Y48在染料废水的处理上具有较大的应用潜能.

基于线粒体细胞色素b基因的细颈囊尾蚴种群遗传多样性研究

为探讨我国细颈囊尾蚴种群之间的遗传相似性及与其他带科带属绦虫的亲缘关系,用线粒体细胞色素b(Cyt b)基因全序列研究了分离自我国四川和云南2省7地区猪、山羊和绵羊共33个细颈囊尾蚴的遗传变异,并用MEGA 4.0程序NJ法和PAUP 4.0程序MP法绘制种系发育树。本研究结果显示:细颈囊尾蚴分离株的Cyt b基因全长为1 068bp,共检测到22个单倍型,单倍型间核苷酸相似性较高(97.3%～99.9%),其与带科带属其他5种绦虫的相似性均在82%以下。系统发育树显示,7个不同地域的细颈囊尾蚴聚为一支,分为3个亚群,且与地理区域、宿主没有直接的相关性,呈现混杂的分布格局,未形成明显的地理分支或宿主分支。研究结果表明:细颈囊尾蚴Cyt b基因存在一定的遗传差异,但种内相对保守,与其他带科绦虫种间差异较大,故可作为种间遗传变异研究的标记,并为细颈囊尾蚴的种群遗传学研究及其与其他带科绦虫病的鉴别诊断的建立奠定基础。

玉米抗矮花叶病毒B株系的QTL定位

为明确玉米对矮花叶病的抗病机制,以代表国内外两大玉米杂种优势类群的优良自交系黄早4和Mo 17为亲本,构建了含239个重组自交系的F9代分离群体,并利用该群体构建了包含101个SSR标记的遗传连锁图谱,图谱全长1422.7 cM,标记间的平均图距为15.6 cM。通过人工接种病毒鉴定,评价了亲本及群体对玉米矮花叶病毒B株系的抗性反应。采用复合区间作图法对玉米矮花叶病抗性QTL进行了定位及其遗传效应分析,在第5、6染色体上,各定位了控制发病率的1个微效QTL和1个主效QTL,分别与标记Bnlg602和Bnlg161连锁,其遗传效应能分别解释表型方差的2.3%和33.8%。

新肽类抗生素B-3543的研究——Ⅰ.产生菌的分类鉴定及生物学特性(英文)

从中国武汉地区水稻白叶枯病叶上分离出一株真菌B-3543,它产生一种新的肽类抗生素,对多种革兰氏阳性和阴性细菌有抗菌活性。其中,对水稻白叶枯病菌抗菌作用最强,其最低抑菌浓度为0.2μg/ml。根据该菌株的形态特征、培养特征和生理特性,鉴定为桃色拟青霉B-3543菌株。

中国被毛孢发酵物对小鼠T、B淋巴细胞转化功能的影响

[目的]观察中国被毛孢发酵物对正常小鼠T、B淋巴细胞转化功能的影响。[方法]给予小鼠高(1.0g·kg-1)、低(0.1g·kg-1)剂量的中国被毛孢发酵物,观察其对小鼠T、B淋巴细胞转化功能的影响。[结果]中国被毛孢发酵物和冬虫夏草均能显著提高正常小鼠脾T和B淋巴细胞转化刺激指数(P<0.05,P<0.01);[结论]中国被毛孢发酵物和冬虫夏草均均能提高正常小鼠免疫功能。