Antibody and T cell responses induced in chickens immunized with avian influenza virus N1 and NP DNA vaccine with chicken IL-15 and IL-18

We had examined the immunogenicity of a series of plasmid DNAs which include neuraminidase (NA) and nucleoprotein (NP) genes from avian influenza virus (AIV). The interleukin-15 (IL-15) and interleukin-18 (IL-18) as genetic adjuvants were used for immunization in combination with the N1 and NP AIV genes. In the first trial, 8 groups of chickens were established with 10 specific-pathogen-free (SPF) chickens per group while, in the second trial 7 SPF chickens per group were used. The overall N1 enzyme-linked immunosorbent assay (ELISA) titer in chickens immunized with the pDis/N1 + pDis/IL-15 was higher compared to the chickens immunized with the pDis/N1 and this suggesting that chicken IL-15 could play a role in enhancing the humoral immune response. Besides that, the chickens that were immunized at 14-day-old (Trial 2) showed a higher N1 antibody titer compared to the chickens that were immunized at 1-day-old (Trial 1). Despite the delayed in NP antibody responses, the chickens co-administrated with IL-15 were able to induce earlier and higher antibody response compared to the pDis/NP and pDis/NP + pDis/IL-18 inoculated groups. The pDis/N1 + pDis/IL-15 inoculated chickens also induced higher CD8+ T cells increase than the pDis/N1 group in both trials (P < 0.05). The flow cytometry results from both trials demonstrated that the pDis/N1 + pDis/IL-18 groups were able to induce CD4+ T cells higher than the pDis/N1 group (P < 0.05). Meanwhile, pDis/N1 + pDis/IL-18 group was able to induce CD8+ T cells higher than the pDis/N1 group (P < 0.05) in Trial 2 only. In the present study, pDis/NP was not significant (P > 0.05) in inducing CD4+ and CD8+ T cells when co-administered with the pDis/IL-18 in both trials in comparison to the pDis/NP. Our data suggest that the pDis/N1 + pDis/IL-15 combination has the potential to be used as a DNA vaccine against AIV in chickens.     

壬基酚对家蚕（Bombyx mori）生长发育的影响

采用添加环境激素壬基酚（Nonylphenol,NP）人工饲料饲养家蚕（Bombyx mori）的验方法,观察了家蚕的生长速度、生长量、变态、生命力情况。结果表明,连续取食0．500mmol·L^-1以上浓度壬基酚的饲料,家蚕幼虫和蛹的生长速度、生长量、发育整齐度、生命力等都明显下降,在发育后期和蜕皮、化蛹、羽化等变态时期更加明显。同时,NP在蚕体内可能有累积作用。2．000mmol·L^-1浓度的NP能使家蚕幼虫很快死亡。NP对家蚕生长发育的影响有明显的性别差异,导致家蚕的雌雄大小开差缩小,并且具有显著的浓度效应和时间效应,是环境激素效应的典型表现。     

Biomass production, foliar and root characteristics and nutrient accumulation in young silver birch (Betula pendula Roth.) stand growing on abandoned agricultural land

The above-ground biomass and production, below-ground biomass, nutrient (NPK) accumulation, fine roots and foliar characteristics of a 8-year-old silver birch (Betula pendula) natural stand, growing on abandoned agricultural land in Estonia, were investigated. Total above-ground biomass and current annual production after eight growing seasons was 31.2 and 11.9 t DM ha⁻¹, respectively. The production of stems accounted for 62.4% and below-ground biomass accounted for 19.2% of the total biomass of the stand. Carbon sequestration in tree biomass reaches roughly 17.5 t C ha⁻¹ during the first 8 years. The biomass of the fine roots (d < 2 mm) was 1.7 ± 0.2 t DM ha⁻¹ and 76.2% of it was located in the 20 cm topsoil layer. The leaf area index (LAI) of the birch stand was estimated as 3.7 m² m⁻² and specific leaf area (SLA) 15.0 ± 0.1 m² kg⁻¹. The impact of the crown layer on SLA was significant as the leaves are markedly thicker in the upper part of the crown compared with the lower part. The short-root specific area (SRA) in the 30 cm topsoil was 182.9 ± 9.5 m² kg⁻¹, specific root length (SRL), root tissue density (RTD) and the number of short-root tips (>95% ectomycorrhizal) per dry mass unit of short roots were 145.3 ± 8.6 m g⁻¹, 58.6 ± 3.0 kg m⁻³ and 103.7 ± 5.5 tips mg⁻¹, respectively. In August the amount of nitrogen, phosphorus and potassium, accumulated in above ground biomass, was 192.6, 25.0 and 56.6 kg ha⁻¹, respectively. The annual flux of N and P retranslocation from the leaves to the other tree parts was 57.2 and 3.7 kg ha⁻¹ yr⁻¹ (55 and 27%), respectively, of which 29.1 kg ha⁻¹ N and 2.8 kg ha⁻¹ P were accumulated in the above-ground part of the stand.     

新城疫病毒NP蛋白MHC I类分子限制性T细胞表位研究

为了对新城疫病毒（Newcastle disease virus,NDV）的细胞毒性T细胞（cytotoxic T cell,CTL）表位进行研究,本研究通过在线分析软件预测了NDV弱毒株La Sota和强毒株Herts/33 NP蛋白的MHC I类分子限制性表位,共获得9条可能的表位多肽序列,并人工合成了这些短肽。分别构建了含有La Sota和Herts/33 NP基因的DNA重组质粒并免疫4周龄SPF级C57BL/6小鼠,首免后21 d加强免疫1次,二免后10 d无菌采集脾脏制备脾淋巴细胞悬液,采用ELISpot法测定预测多肽诱导脾淋巴细胞分泌IFN-γ的能力,根据分泌IFN-γ形成的特异性斑点数进行生物统计学分析,发现2条多肽产生的斑点数显著高于无关肽刺激组（P＜0.01）。确定了针对小鼠H-2 kb的限制性T细胞抗原表位的2条多肽。这些可用于合成特异性的MHC-肽四聚体,为后续研究NDV与树突状细胞（dendritic cells,DC）及T淋巴细胞之间的免疫抑制机制提供重要保障。     

H9N2亚型禽流感病毒NP蛋白原核表达及多克隆抗体制备

【目的】对华南地区分离到的1株H9N2亚型禽流感病毒(Avian influenza virus, AIV)流行株NP蛋白进行原核表达,制备H9N2亚型AIV NP蛋白多克隆抗体。【方法】将1株H9N2亚型AIV的NP基因克隆至pET-32a(+)原核表达载体,构建NP蛋白原核表达质粒。将重组质粒同时转化大肠杆菌BL21(DE3)和Rosetta(DE3)感受态细胞,经IPTG诱导表达重组蛋白。通过考马斯亮蓝染色和Western blotting分析并比较NP蛋白在两种表达菌中的表达量,进一步优化诱导剂浓度、诱导时间及诱导温度等条件,提高蛋白的表达效率。采用镍柱亲和层析法对NP蛋白进行纯化,用BCA法测定蛋白浓度。以纯化的NP蛋白免疫新西兰大白兔获得多克隆抗体,通过Western blotting和间接免疫荧光对所制备的多克隆抗体进行鉴定,通过间接ELISA测定抗体效价。【结果】试验成功构建pET-32a-H9N2-NP原核表达质粒并表达重组NP蛋白。考马斯亮蓝染色和Western blotting结果均表明,重组NP蛋白在大肠杆菌Rosetta(DE3)感受态细胞中表达水平远高于大肠...     

普通念珠藻多糖复合物对植物生长的促进作用

以普通念珠藻为原料,用水作提取剂,提取得到普通念珠藻多糖复合物N(P)。以水稻、番茄、白菜上海青、吊钟花、墨兰、柳枝为材料,研究NP对试验对象生长的影响。结果表明,NP能促进盆栽水稻的生长和分蘖;使盆栽番茄株高增高,株茎增粗,鲜重、干重增加;使盆栽上海青白菜鲜重、干重增加;促进盆栽吊钟花的株高增高,花芽数增多;使盆栽墨兰分蘖率提高;促进柳枝根系生长,新芽数增多,且芽长增长;促进大田番茄株高增高、株茎增粗,增加成熟果总重。     

一株Class Ⅰ新城疫病毒分离株NP和P蛋白基因分子特征和遗传进化分析

本研究对2009年实验室保存的一株Class Ⅰ新城疫病毒分离株NDV09-056的NP和P基因进行了扩增和序列分析,对其分子特征和遗传进化关系进行了分析。结果表明该分离株NP蛋白基因全长为1746nt,共编码489aa,其N端1～401氨基酸相对比较保守,C端402～479氨基酸变化比较大;同源性分析表明本分离株的NP蛋白基因与Ⅰ类新城疫病毒代表毒株之间核苷酸的同源性为93.5%～98.5%,而与Ⅱ类新城疫病毒代表毒株的同源性较低,介于77.0%～79.2%。P蛋白主要氨基酸突变区集中在57～107aa和135～212aa两个区域,而N端和C端相对保守;同源性分析表明本分离株的P蛋白基因与Ⅰ类新城疫病毒代表毒株之间核苷酸的同源性为90.4%～95.3%,而与Ⅱ类新城疫病毒代表毒株的同源性较低,介于68.3%～72.5%。遗传进化分析表明本分离株与Ⅰ类基因3型新城疫病毒代表株NDV08-004的亲缘关系最近。     

复方硝基酚钠诱发小鼠骨髓细胞和睾丸精原细胞染色体畸变的观察

研究了鱼虾促生长药物添加剂复方硝基酚钠对体细胞和雄性生殖细胞的致突变性。骨髓细胞染色体畸变分析以每公斤体重用２５０、８３．３、２５ｍｇ的复方硝基酚钠分别给小鼠经口染毒，每日１次，连续５ｄ，同时设空白对照组和阳性对照组。睾丸精原细胞染色体畸变分析以每公斤体重用２５０、１２５、６２．５ｍｇ的复方硝基酚钠给雄性小鼠经口染毒，每日１次，连续５ｄ，同时设空白对照组和阳性对照组。上述两种致突变试验结果均为阴性。因此认为复方硝基酚钠对哺乳动物体细胞和雄性生殖细胞均未显示致突变作用     

Effect of Integrated Nutrient Management on Soil Fertility and Yield of Rice (Oryza sativa L.) under the System of Rice Intensification in the Indian Subtropics

Field experiments were conducted at Water Management Research Station, Begopara, Nadia, WB, India, during the rabi seasons of 2008–2009 and 2009–2010 to find out the integrated effect of nitrogen (N), phosphorus (P), potassium (K), farmyard manure (FYM) and zinc (Zn) under the system of rice intensification (SRI) techniques using eight treatments on the fertility changes in soil. The results revealed that the amounts of organic carbon and available N content in soil were found to maintain the highest fertility status with the highest yield in T₆ (NPK + FYM 10 tha^?1 + Zn 5 kgha^?1) and gave the highest N uptake (55.98 kgha^?1). The availability of P decreased with the increased level of Zn application and gave the highest P uptake (23.52 kgha^?1) in the treatment T₅ (NPK + FYM 10 tha^?1). The highest Zn content (4.71 mgkg^?1) was recorded in the treatment T₇ (NPK + FYM 10 tha^?1+ Zn 10 kgha^?1).     

Genetic characterization of influenza A viruses circulating in pigs and isolated in north-east Spain during the period 2006–2007

Swine influenza virus is one of the most important pathogens involved in the swine respiratory disease complex. Recent serological surveys showed a high prevalence of swine influenza strains belonging to the H₁N₁, H₁N₂ and H₃N₂ subtypes circulating in pigs in Spain. However, little is known about their genome sequence. Five swine influenza strains were isolated from some unrelated outbreaks occurred during 2006–2007, and their complete genome sequences were determined. Phylogenetic analysis revealed that they belonged to the lineages “Avian-Like” H₁N₁, “Human-Like” H₃N₂, and “Human-Like” H₁N₂, showing tight relationships with early or contemporary strains described in Europe. Notably, one virus of the H₁N₂ subtype showed genetic and antigenic divergence with the European contemporary strains or vaccinal strains of the same subtype, suggesting that some local and divergent clusters of the virus may pass unnoticed in routinary subtyping. Finally, analysis on the entire pattern of genome segments suggested that a second reassortment event could have influenced the evolution of that divergent H₁N₂ strain.