A dominant antigenic epitope on SARS-CoV spike protein identified by an avian single-chain variable fragment (scFv)-expressing phage

Severe acute respiratory syndrome (SARS) is a newly emergent human disease, which requires rapid diagnosis and effective therapy. Among antibody sources, immunoglobulin Y (IgY) is the major antibody found in chicken eggs and can be used as an alternative to mammalian antibodies normally used in research and immunotherapy. In this study, phage-expressing chicken monoclonal scFv antibody was chosen and characterized with phage display antibody technology. Truncated fragments of SARS-CoV spike protein were cloned in pET-21 vector and expressed in BL-21 Escherichia coli (E. coli) cells. After purification, the purity of these recombinant spike proteins was examined on SDS-PAGE and their identity verified with Western blot analysis using anti-his antibodies and sera from convalescent stage SARS-CoV-infected patients. Using these bacteria-derived proteins to immunize chickens, it was found that polyclonal IgY antibodies in the egg yolk and sera were highly reactive to the immunogens, as shown by Western blot and immunocytochemical staining analysis. A phage displaying scFv library was also established from spleen B cells of immunized chicken with 5 x 10(7) clones. After four panning cycles, the eluted phage titer showed a 10-fold increase. In sequence analysis with chicken germline gene, five phage clones reacted, with large dissimilarities of between 31 and 62%, in the complementarity-determining regions, one dominant phage 4S1 had strong binding to fragment Se-e, located between amino acid residues 456-650 of the spike protein and this particular phage had significantly strong binding to SARS-CoV-infected Vero E6 cells. Based on the results, we conclude that generating specific scFv-expressing phage binders with the phage display system can be successfully achieved and that this knowledge can be applied in clinical or academic research.     

Effects of inulin on performance, egg quality, gut microflora and serum and yolk cholesterol in laying hens

1. The present study was conducted to investigate the effects of inulin on laying hens. A total of 360 Brown Nick laying hens were divided randomly into 6 groups of 60 with 6 replicates of 10 hens and fed on diets containing 0 (control), 0·1, 0·5, 1·0, 1·5 or 2·0% inulin during the 4-week trial. 2. Dietary supplementation of inulin reduced cholesterol concentration (mg/g yolk) and content (mg/egg) in eggs. Cholesterol content in eggs decreased linearly with increasing levels of dietary inulin level. 3. Supplementation of inulin in diets decreased coliform bacteria counts and pH in the caecum. The lowest coliform bacteria counts (6·30 ± 0·03 log10 cfu/g) and pH (6·47 ± 0·01) were obtained in the 2·0% inulin group, the two indices decreasing by 21·6% and 3·0% respectively, compared with the control group. Coliform bacteria count and pH were changed linearly in accordance with increasing levels of dietary inulin level. Caecal Bifidobacteria counts were increased in the 2·0%-inulin group. 4. Inulin supplementation of layer diets did not appear to have any adverse effects on laying rate, egg weight, feed intake, feed conversion efficiency, cracked-egg rate, eggshell thickness or Haugh unit compared with the control laying hens. 5. Therefore, dietary supplementation with inulin may lead to the development of low-cholesterol chicken eggs as demanded by health-conscious consumers.     

Hematology values from clinically healthy Peromyscus leucopus.

Reported herein are hematocrit and total and differential WBCs obtained from 132 clinically healthy male and female white-footed mice (Peromyscus leucopus) of four different age groups: 4, 5-8, 9-20, and >32 wk old. Minimal differences were identified between the age groups. The number and percentage of neutrophils in the 4-wk-old mice were significantly higher than those in the other groups; the relative percentage of lymphocytes in the 4-wk-old mice was significantly lower than in the 9-20-wk-old mice. The only significant gender effects identified were higher numbers of WBCs and lymphocytes in females of the 4-wk-old group and higher hematocrits in males of the 5-8- and >32-wk-old groups.     

平武黄牛体尺外貌的研究

对平武黄牛主产区生态条件的调查、外貌的观察和体尺的测定,与四川现有黄牛地方品种的比较分析,结果表明平武黄牛在体尺、外貌、体尺指数等方面有别于其它黄牛品种,是川西北山区良好的役用型黄牛种群,具有向役肉兼用型方向发展的优良特性。     

桑椹红色素稳定性的研究

研究了光和热等物理因素、常用食品添加剂、金属离子、氧化剂及还原剂对桑椹红色素稳定性的影响。结果表明 :pH值对桑椹色素的影响显著 ,在酸性条件下色素稳定 ;长时间高温和光照对色素有明显的破坏作用 ;金属离子Na+ 、Ca2 + 、Cu2 + 、Fe2 + 、Mg2 + 、Zn2 + 和Mn2 + 对桑椹色素色泽有一定的增色效果 ,但Fe3 + 则有明显的不良影响 ;食品中常用的葡萄糖、蔗糖、苯甲酸钠和山梨酸钾对桑椹色素无明显的不良影响 ;VC对桑椹色素有双重作用 ,而H2 O2 和Na2 SO3 对色素有严重的破坏作用。     

肉兔不同杂交组合的生产性能测定

养兔具有投资少、见效快、效益高的特点，饲养规模可大可小，饲养方式多种多样，是当前广大农村实现农业增效、农民增收的好项目。但是，养兔效益可受品种、繁育方式、饲料营养、饲养管理等多种因素的影响。据生产实践，作为商品兔，一般杂种兔的经济效益高于纯种兔，杂种后代的增重速度可比纯种兔提高10％以上。但不同品种间杂交，其杂交效果各不一样。     

重要营养素--血红素铁

血红素铁是卟啉类化合物,是哺乳动物血液中的天然色素和生物铁,广泛应用于医药、食品和化工业。文章就血红素铁的理化特性、生理功能及应用进行了简述,并对其常用提取方法进行了比较概述。     

草地畜牧业是推动黔西南州农业结构战略牲调整的突破口

改革开放20多年来。黔西南州农业获得了长足发展，在产品供求关系、生产目标、增长方式、经营形式、产业关联等方面都发生了显著变化，进入了农产品供求基本平衡，以提高品质、优化结构和增加农民收入为主，注重传统投入与资本集约和技术集约相结合的优化发展阶段。加入WTO，对于我州正处在发展新阶段的农业和农村经济来说。是一次重要的机遇和挑战。面对激烈的国际、国内市场竞争。在西部大开发中，为了改善生态环境，调整农业结构，增加农民收入，州委、州政府决定把草地畜牧业作为我州农业和农村产业结构调整的重点。制定了百万亩草地畜牧业发展规划。出台了加快草地畜牧业发展的实施意见。     

应用兔肾原代细胞培养 兔粘液瘤病毒的研究

试用兔肾原代细胞传代；增殖兔粘液瘤病毒，呈现典型ＣＰＥ。粘液瘤病毒ＳＧ３３株在兔肾细胞上连续传３代之后，就不产生ＣＰＥ，在鸡胚上传３代再回归兔肾原代细胞，又呈现ＣＰＥ。ＳＧ＿（３３）在鸡胚上不能产生明显痘斑。维持液ｐＨ值影响ＳＧ＿（３３）在兔肾原代细胞上形成ＣＰＥ。     

两种蔗糖浓度对切割取样鼠胚冷冻解冻效果的研究

小鼠晚期桑椹胚、早期囊胚和扩大囊胚，经切割取样分别采用０．３６５Ｍ和０．２５Ｍ蔗糖浓度进一步细管和分步脱除甘油解冻后体外培养发育率分别为６５．５％、４６．４％和７５．４％，两种浓度间差异显著（Ｐ＜０．０５）；分步脱除甘油解冻瑟前者差异不显著（Ｐ＞０．０５），与后者差异极显著（Ｐ＜０．０１）。