Expression of Anti-Lipopolysaccharide Factor Isoform 3 in Chlamydomonas reinhardtii Showing High Antimicrobial Activity

Antimicrobial peptides are a class of proteins with antibacterial functions. In this study, the anti-lipopolysaccharide factor isoform 3 gene (ALFPm3), encoding an antimicrobial peptide from Penaeus monodon with a super activity was expressed in Chlamydomonas reinhardtii, which would develop a microalga strain that can be used for the antimicrobial peptide production. To construct the expression cluster, namely pH2A-Pm3, the codon optimized ALFPm3 gene was fused with the ble reporter by 2A peptide and inserted into pH124 vector. The glass-bead method was performed to transform pH2A-Pm3 into C. reinhardtii CC-849. In addition to 8 μg/mL zeocin resistance selection, the C. reinhardtii transformants were further confirmed by genomic PCR and RT-PCR. Western blot analysis showed that the C. reinhardtii-derived ALFPm3 (cALFPm3) was successfully expressed in C. reinhardtii transformants and accounted for 0.35% of the total soluble protein (TSP). Furthermore, the results of antibacterial assay revealed that the cALFPm3 could significantly inhibit the growth of a variety of bacteria, including both Gram-negative bacteria and Gram-positive bacteria at a concentration of 0.77 μM. Especially, the inhibition could last longer than 24 h, which performed better than ampicillin. Hence, this study successfully developed a transgenic C. reinhardtii strain, which can produce the active ALFPm3 driven from P. monodon, providing a potential strategy to use C. reinhardtii as the cell factory to produce antimicrobial peptides.     

护林防火技术在森林资源保护中的应用

合理运用护林防火技术可以提升森林资源保护工作的整体质量,推动林业不断发展。本文分析了森林火灾发生的原因,总结了护林防火技术,提出了相关应用对策,以期为减少森林火灾的发生和保护森林资源提供参考。     

Soil protection for a sustainable future: options for a soil monitoring network for Ireland

The increased recognition of the importance of soil is reflected in the UN Post‐2015 Development Agenda with sustainable development goals that directly and indirectly relate to soil quality and protection. Despite a lack of legally binding legislation for soil protection, the European Commission remains committed to the objective of soil protection. However, the achievement of a legally binding framework for soil protection relies on the implementation of a soil monitoring network (SMN) that can detect changes to soil quality over time. As beneficiaries do not pay for the provision of soil information, the options for soil monitoring are limited. The use of existing data sets should be considered first. Using Ireland as an example, this research explored the opportunities for a SMN for Ireland considering three existing national data sets. The options for a SMN are considered in terms of their spatial and stratified distribution, the parameters to be measured and an economic analysis of the options proposed. This research finds that for Ireland, either a 10 or a 16 km² grid interval stratified by land use and drainage class offers the best potential in relation to the spatial distribution of existing data sets to reflect local data at a national level. With existing data, the stratified SIS data using the 16 km² grid offers the best value for money, with baseline costs for analysis, excluding field costs, of between €706 481 and €2.8 million. Acknowledging the impossibility of measuring all parameters with ideal frequency, this study proposes a two‐tier system for optimized monitoring frequency. Parameters must anticipate future policy requirements. Finally, the implementation of a SMN must be accompanied by standardized methods, defined thresholds and action mandates to maintain soil quality within allowable limits.     

社会林业与五指山的自然保护

五指山自然保护区是世界上现存地势最高的原始热带雨林之一，是我省目前面积最大的自然保护区，也是海南三大河流的发源地。但由于保护区横跨琼中、通什、保亭三县、市，紧邻保护区还生存着众多贫困的黎、苗族村民。因此，保护区与周边社区的联系是十分紧密的，社区的发展也直接影响到五指山热带天然林的保护。海南省林业局中德海南热带林保护和恢复项目在社会林业与五指山森林的保护上做出了有益的尝试，就在全省来说，也是首次对社会林业与森林保护上进行的探索。五指山地区社会林业的研究，对保护海南热带雨林也有着积极的意义。特对社会林业与五指山的森林保护进行了阐述、分析和设想。     

The Brief Introduction to the Sino-US Joint Centers for Soil and Water Conservation and Environmental Protection

Erosion and transport of soil has worldwide implications for agriculture, landscape stability, climate, natural hazards, and clean, renewable resources of water and air. Assured access to clean water and a healthy and safe environment requires an ethic of conservation and protection. The minimum scale in which these principles apply successfully is basin wide. These are the fundamental concerns of the Sino-US Centers for Soil and Water Conservation and Environmental Protection.     

Passive protection of mice with antiserum to neuraminidase fromPasteurella multocida serotype A:3

Antiserum to a partially purified neuraminidase fromPasteurella multocida, type A:3, was adsorbed with protease-digestedP. multocida type 3 lipopolysaccharide (LPS) to remove LPS immunoreactivity. The LPS-adsorbed antineuraminidase caused a 77% reduction in the neuraminidase activity of homologousP. multocida in anin vitro enzyme neutralization test. All 14 mice passively immunized with the adsorbed antineuraminidase were protected against challenge infection with homologousP. multocida in a mouse protection test. Ten out of 14 mice in one group that received antisera containing antibodies to both neuraminidase and LPS were protected. In contrast, only 1 out of 14 mice that were immunized with pre-immune serum survived the challenge. These results suggest that antiserum toP. multocida neuraminidase was, at least partly, responsible for the protection observed in this study. Neuraminidase may be one of the immunogenic protective proteins present in aqueous extracts ofPasteurella multocida.     

金属热防护系统多层隔热材料的传热分析及参数优化

采用能量平衡方程和离散坐标法近似地建立了金属热防护系统多层隔热材料的一维稳态传热的数学模型，并利用遗传算法和纤维隔热材料的有效导热系数试验数据求解传热反问题，从而得到了模型中的纤维隔热材料的辐射衰减系数、反照率和隔热屏表面辐射发射系数。最后，由实验测得了多层隔热材料的有效导热系数，验证了采用优化参数后，多层隔热材料的传热模型的正确性。     

马传贫驴白细胞弱毒疫苗株跨膜蛋白主要免疫决定区基因的克隆与表达

从感染驴白细胞的马传贫驴白细胞弱毒疫苗株前病毒DNA中克隆了编码跨膜蛋白主要免疫决定区(TMIR)的基因，并在大肠杆菌中进行了表达。所表达的融合蛋白有一部分是可溶的，其氨基端带有6个组氨酸的标签，因此可以用固定化金属离子亲和层析法在非变性条件下进行纯化。在间接酶联免疫吸附试验(ELISA)和免疫印迹试验中，重组的TMIR蛋白可与马传贫阳性血清样品发生反应，而与健康马血清无任何反应。这表明该重组蛋白具有良好的抗原性和特异性，可用于马传贫弱毒疫苗株在体内外复制、接种马体内免疫应答及马传贫诊断的研究。     

鸡致病性大肠杆菌I型菌毛交叉保护性研究及其结构基因 FimA 的序列分析

从山东省各地分离到107株鸡致病性大肠杆菌，选择了其中2株高致病性大肠杆菌，血清型分别为O78和OM，经增菌培养后提取菌毛制备为单价菌毛油乳苗，分别接种于1日龄和14日龄雏鸡，于4周龄攻毒。结果二者之间有一定的交叉保护作用。根据Genbank收录的人源大肠杆菌I型菌毛FimA基因和P型菌毛papA基因序列，分别设计了2对引物。通过PCR对上述2株大肠杆菌进行扩增，结果只有FimA的一对引物扩增出相应的条带，经测序证明为FimA基因。papA基因的引物未扩出任何条带，证明这2株大肠杆菌表达I型菌毛。通过对2株大肠杆菌结构基因FimA进行分析，发现二者具有高度的同源性。本研究的目的是探讨菌毛亚单位之间的交叉保护性与其菌毛的结构基因之间是否存在相关性。     

鸡空肠弯曲菌灭活苗的制备及其免疫保护试验研究

用分离到的空肠弯曲菌制备鸡空肠弯曲菌甲醛灭活疫苗 ,并进行了该病的免疫保护实验。制备 2× 10 9个 / m L浓度的细菌悬液 ,4m L / L浓度的甲醛溶液作用 12 h灭活后制成空肠弯曲菌灭活苗 ,免疫 2周龄的 SPF雏鸡 ,14 d后 ,进行攻击实验 ,观察其临床症状 ;并分别于感染后 7d和 14 d捕杀 ,观察病理学变化及进行细菌分离实验以揭示其保护率。结果表明 ,鸡空肠弯曲菌灭活苗的保护率可达 10 0 % ,说明该苗经口免疫后安全可靠 ,无毒副作用