Validation of RT‐PCR Assays for Molecular Characterization of Porcine Teschoviruses and Enteroviruses

Porcine enteroviruses (PEVs) and teschoviruses (PTVs) are described as causative agents of neurological disorders, fertility disorders and dermal lesions of swine. Difficulties in the serological detection of these viruses may lead to a significant underestimation of infections with clinical symptoms. With the recent availability of genome sequence data for all the serotypes, molecular diagnosis is a possibility. The present study describes a new approach to molecular ‘serotyping’ of PTVs and PEV‐B viruses, involving the amplification and sequencing of a genomic fragment of the VP1 coding region. A molecular characterization of Italian entero‐teschovirus isolates was performed using a set of previously published and newly designed polymerase chain reaction primers. A total of 33 porcine isolates and 10 reference strains were analysed. Porcine enterovirus‐B samples were first diagnosed as positive for enterovirus by amplification of the 5′‐non‐translated region. Samples were then typed by amplification and sequencing of a portion of the VP1 coding region. Porcine enterovirus‐A and PTVs were detected by a published assay in the 5′‐NC region that allows them to be differentiated according to the size of amplification product, using the same set of primers. For serotype characterization of PTV, we evaluated four different regions: the N terminus of the capsid protein VP2, the region encoding for RNA‐dependent RNA polymerase, and the capsid VP1 and VP4 regions. The newly designed primers in the VP1 region was proved to be broad in range and suitable for serotype assessment and therefore constitute a useful diagnostic tool for molecular diagnosis of porcine teschovirus/enterovirus strains and for the study of molecular epidemiology and evolution of these viruses.     

Synthesis and insecticidal activities of pyridine ring derivatives of podophyllotoxin

In an attempt to find the biorational pesticides, we synthesized 12 pyridinyl derivatives of podophyllotoxin (PPT) and 4′-demthylepipodophylltoxin (4′-DMEP) in this study. Their structures and the α/β substitution at C-4 were confirmed by ¹H NMR, IR, MS spectral analyses and elemental analysis. The insecticidal activities were tested against fifth-instar larvae of Pieris rapae and the third-instar larvae of Cullex pipiens pallens at concentrations of 250 and 10 μg ml⁻¹. Four derivatives of PPT, 4.1, 4.2, 4.3 and 4.5, showed higher insecticidal activities against P. rapae than PPT, while three derivatives of PPT, 4.4, 4.5 and 4.6, displayed higher mosquito larvicidal activity than PPT, with LC₅₀ values of 1.66, 3.96 and 1.54 mg l⁻¹, respectively. Interestingly, we also found that the pyridine ring derivatives of PPT showed delayed insecticidal activity, which is different from traditional neurotic insecticides. The results suggest that 4′-OCH₃ in the PPT derivatives is essential to keep the insecticidal activity and the insecticidal activities of pyridine ring derivatives of PPT are higher than that of the derivatives of 4′-DMEP, supporting PPT has the potential to be a lead structure of semi-synthetic insecticides.     

表达猪繁殖与呼吸综合征病毒CH-1a株核衣壳蛋白重组鸡痘病毒的构建

从含有猪繁殖与呼吸综合征病毒（PRRSV）核衣壳蛋白（N）的质粒扩增出N基因，构建禽痘病毒转移载体。该载体含有禽痘病毒早晚期启动子LP2EP2控制之下的PRRSVN基因、P11启动下的报告基因lacZ以及用于同源重组的禽痘病毒基因组的片段。在转移载体转染亲本病毒S—FPV-017感染的鸡胚成纤维细胞（CEF）之后，采用蓝色表型筛选的方法，筛选到表达N基因的重组病毒，并对其进行了6轮蚀斑纯化。PCR方法鉴定证明重组病毒的基因组中含有完整PRRSVN基因，间接免疫荧光试验证明了PRRSVN蛋白在重组病毒感染的CEF细胞中获得表达，本研究为猪繁殖与呼吸综合征非复制型疫苗的研制打下了基础。     

20%咪鲜胺·异菌脲悬浮剂的高效液相色谱分析

本文采用高效液相色谱法分析20%咪鲜胺·异菌脲悬浮剂,使用C18反相柱和紫外可变波长检测器,以甲醇 水为流动相,用外标法对有效成分进行分析和定量。咪鲜胺和异菌脲标准偏差分别为0.055和0.021变异系数分别为0.53%和0.19%。平均回收率分别为98.95%和99.10%。     

Leaf necrosis and twig dieback of sweet persimmon (Dyospiros kaki L.) caused by Pseudomonas syringae pv. syringae in Italy

In spring 1996, extensive leaf necrosis and twig dieback were observed on young sweet persimmon (Dyospiros kaki L.) trees, cultivars O'Gosho, Hachija, Mercatelli and Kaki-tipo planted in the Abruzzo region (central Italy). Many trees were killed. When the dieback reached the trunk, in many cases, new vegetation was noticed above the graft point. The cultivar Jiro-C was not affected by the disease. During 1997, no symptoms were observed on any plant. The orchard was planted in a clay soil with a very low content of organic matter. Biochemical, nutritional and pathogenicity tests indicated Pseudomonas syringae pv. syringae van Hall as the causal agent of the disease. This is the first report of this bacterium as a pathogen of sweet persimmon in Europe.     

我国复配食品添加剂发展现状、问题及对策

综述复配食品添加剂的定义、分类和特点,针对我国复配食品添加剂在生产应用中存在的主要问题,提出相应的解决措施,展望复配食品添加剂在我国的发展前景,为推动复配食品添加剂产业发展提供借鉴。     

耦合水文情势及鱼类繁殖的江垭水库生态调度需求研究（生态调度专刊）

大型水库实施生态调度是修复河流水文情势、维持河流生态系统健康的重要手段。江垭水库位于溇水干流下游的张家界慈利县，是澧水流域具有重要防洪意义的大型水库。本研究通过环境DNA采样、IHA水文指标变动分析和文献资料总结等方法，开展了江垭坝下水文情势及鱼类繁殖需求的关系研究，提出了江垭水库生态调度需求及建议。2021年9月在溇水干支流综合调查到鱼类55种，与上世纪90年代相比，流水性种类在干流江段显著减少了51.35%；按照生态调度优先等级评估原则，确定江垭水库生态调度的主要目标物种为产漂流性卵的银鲴、银鮈、贝氏?和产粘沉性卵的鲤、鲫；江垭建坝前后下游水文情势变化较大，发生高度改变且对鱼类洄游产卵及鱼卵发育有重要作用的水文指标包括：年最大1日、3日平均流量指标，年出现高流量脉冲事件的次数和持续时间，以及连续日流量上涨率。以促进坝下不同产卵类型的鱼类繁殖为目标，江垭水库生态调度一方面需要恢复一定的高流量脉冲次数和历时，另一方面需要控制下游的水位日降幅，通过泄放合理的生态流量来维持下游鱼类的生物多样性。     

变量筛选方法对郁闭度遥感估测模型的影响比较

比较基于偏最小二乘回归的Bootstrap方法与传统的平均残差平方和(RMSq)准则所选变量建立模型的精度差别.结果表明:Bootstrap方法是一种更优秀的变量筛选方法,比RMSq方法精度提高约5%;而且它不受变量多带来的运算困难的限制,更便于实际应用.