Immune responses of orange-spotted grouper,Epinephelus coioides,against virus-like particles of betanodavirus produced in Escherichia coli

Betanodaviruses are the causative agents of viral nervous necrosis (VNN), a serious disease of cultured marine fish worldwide. Virus-like particles (VLPs) are one of the good novel vaccine candidates to control this disease. Until now, betanodavirus vaccine studies mainly focused on the humoral immune response and mortality after virus challenge. However, little is known about the activation of genes responsible for cellular and innate immunity by vaccines. In the present study, VLPs of orange-spotted grouper nervous necrosis virus (OGNNV) were produced in prokaryotes and their ability to enter Asian sea bass cells was the same as native virus, suggesting that they possess a similar structure to OGNNV. VLPs immunogenicity was then determined by intramuscularly vaccinating Epinephelus coioides at different concentrations (1.5 or 15 μg g^?1 fish body weight, FBW) and immunizing frequencies (administration once, twice and thrice). A single vaccination with the dosage of 1.5 μg g^?1 FBW is enough to provoke high titer antibodies (average 3 fold higher than that of negative control) with strong neutralizing antibody titer as early as 1 week post immunization. Furthermore, quantitative PCR analysis revealed that eleven genes associated with humoral, cellular and innate immunities were up-regulated in the liver, spleen and head kidney at 12 h post immunization, correlating with the early antibody response. In conclusion, we demonstrated that VLP vaccination induced humoral immune responses and activated genes associated with cellular and innate immunity against betanodavirus infection in orange-spotted grouper.     

犬细小病毒抗体间接ELISA检测方法的建立

利用本实验室建立的昆虫细胞/杆状病毒表达系统表达犬细小病毒病毒样颗粒（CPV-VLPs）,采用硫酸铵沉淀、蔗糖密度梯度离心对表达的CPV-VLPs进行纯化,用电子显微镜、SDS-PAGE及Western-blotting方法检测纯化效果。以纯化的CPV-VLPs作为包被抗原建立CPV间接ELISA检测方法,对各反应条件进行优化并分析其特异性、敏感性、重复性。结果显示,CPV-VLPs经过纯化后纯度可达到95%以上;优化的ELISA最佳工作条件为：纯化抗原包被浓度为5.0mg/L,4℃包被过夜;1%BSA,37℃封闭2h;待检血清1∶40稀释,37℃孵育1.5h;HRP标记的酶标二抗1∶20 000稀释,37℃孵育1h;TMB室温避光显色30min;确定的血清阴性阳性临界值D450nm值为0.264。该方法可特异性检测犬细小病毒阳性血清,与犬瘟热、犬传染性肝炎、犬冠状病毒病、狂犬病阳性血清均不发生反应。该方法的敏感性为1∶640,批内重复试验变异系数小于6%,批间重复试验变异系数小于8%。42份临床血清样本的检测结果表明,与血凝抑制试验的符合率为90.48%。     

Cloning and sequence analysis of nine novel MYB genes in Taxodiaceae plants

Myeloblastosis(MYB) is one of the largest transcribed factor families in plants. To gain an overall picture of the evolution of MYB genes in relict plants, we cloned nine novel MYB genes in Taxodiaceae plants(Taxodium distichum, Taxodium ascendens, Cryptomeria japonica var. Sinensis, Cryptomeria japonica cv. Araucarioides, Cryptomer Japonica, Metasequoia glyptostroboides, Cunninghamia lanceolata, Taiwania cryptomerioides and Glyptostrobus pensilis). The deduced amino acid sequences for MYBs showed that the nine MYB proteins contained two DNA binding domains. The first domain is from amino acid position 29 to 78, wherein three tryptophanes at 33, 53 and 73 were separated by 19 amino acids, respectively. The second domain is from amino acid position 82 to 127, wherein three tryptophanes at 86, 105 and 124 were separated by 18 amino acids, respectively, whereas the first tryptophane at amino acid position 86 is replaced by a phenylalanine. The characterization of these conserved domains at nine MYBs indicated that they all belong to the R2R3-MYB group. The secondary structure analysis showed that α-helix and β-turn are the major motifs of the predicted secondary structure of MYBs. The three dimensional model of each MYB protein showed that the structure is like clip, making it more flexible and mobile. The similarities between the nine MYB proteins in Taxodiaceae were calculated. The highest identical value of 99% is between CjsMYB, CjMYB and CjaMYB, whereas the lowest value of 82% is between TaMYB and ClMYB. According to the phylogenetic tree, the distances between different genera were relatively large whereas those within genera were relatively small. As expected, accessions of the same genus formed a subgroup before being grouped with other genera.     

会稽山古香榧群传统农业动态保护途径探讨

香榧是古老的中国原产树种和世界稀有的经济树种,具有果用、药用、油用、观赏等众多功能。会稽山区是香榧的原产地和主产地,其千年古香榧群是一种重要的农业文化遗产。在系统分析会稽山古香榧群特点基础上,提出了会稽山古香榧群传统农业保护的重要意义及动态保护的主要途径。研究对于保护好会稽山千年古香榧群,合理利用其优良种质资源,促进古香榧群传统农业的继承与发展具有一定的指导意义。     

饲料粉碎粒度和调质温度对肉鸡养分消化率及生长性能的影响

饲料制粒可以提高采食量和缩短采食时间,以提高肉鸡生长性能和饲料转化率.而原料的粉碎和调质是制粒过程中非常重要的环节,不仅直接影响到饲料的营养价值和肉鸡的消化吸收,还会影响加工成本和产品质量.粉碎粒度过大会导致混合不均匀、颗粒品质欠佳等不利影响,并且对动物的采食及采食后的物理性消化产生影响,粒度过小则会对动物健康产生不利影响.调质过程主要是使饲料受到水热作用,使其蛋白质变性、淀粉糊化等,调质温度是调质环节的关键参数,温度的高低对蛋白质变性及淀粉糊化程度有直接影响,从而间接影响到动物的生长性能和养分消化利用率.同时,调质温度过高会导致饲料中添加的酶制剂及维生素等严重失活,反而会降低饲料的营养价值.本文综述了肉鸡饲料粉碎粒度和调质温度的研究进展,阐述了粉碎粒度、调质温度与肉鸡生长性能和养分消化率的重要关系.     

养殖环境细菌微生物气溶胶粒径分布及其健康危害评估

为深入了解畜禽舍环境中气载细菌微生物的空气动力学粒径分布规律,并评估其潜在的健康危害风险,采用Andersen-6级微生物空气采样器以血-琼脂培养基、沙氏培养基和高氏合成1号培养基为采样介质,对鸡舍、猪舍、牛舍环境中空气样品进行系统定点取样、测定及分析。研究结果表明,鸡舍环境中气载需氧菌含量最高,猪舍次之,牛舍最低;空气细菌粒径分布均为第Ⅰ级最高,鸡舍空气粒径呈偏态分布,牛舍、猪舍分别在第Ⅲ级和第Ⅳ级出现第2个峰值。携带细菌可吸入微粒在猪舍环境中比例最大。空气真菌与放线菌均在第Ⅳ级最高,携带真菌和放线菌可吸入粒子的比例显著大于细菌（P<0.05）。鸡舍、猪舍、牛舍空气微生物粒径各级分布比例基本一致。在鸡舍、猪舍、牛舍每天约有6.1×105CFU、4.7×104CFU和3.6×104CFU气载细菌微生物可分别进入人和动物小支气管或直接进入肺泡,从而对人和动物健康构成潜在危害。     

泉州山区乡镇组团发展现代农业的调查与思考

山区乡镇组团发展现代农业,旨在强化同一县(市、区)内或不同县(市、区)相邻乡镇之间的合作共赢。为深入了解山区乡镇发展现状,促进山区乡镇组团发展现代农业,课题组在泉州南安市罗东镇、九都镇、乐峰镇、向阳乡和永春县外山乡、洛江区虹山乡展开调研,梳理归纳出该区域组团发展现代农业的现实条件与制约因素,并对山区乡镇组团发展现代农业的运作思路提出合理化建议。     

Influence of high-mobility group box 1 on proliferation of neural stem cell in pre-infarction cortex of focal cerebral ischemia/reperfusion model rats

AIM：To investigate the influence of high-mobility group box 1 (HMGB1) on the proliferation of neural stem cells in peri-infarction cortex of focal cerebral ischemia/reperfusion model rats. METHODS：Male SD rats (n=48) were randomly divided into sham group, ischemia/reperfusion (I/R) group, RNA interference group and negative interference group. The rat middle cerebral artery was blocked to establish focal cerebral I/R model (ischemia for 1 h and reperfusion for 7 d). Lentivirus vector of HMGB1 shRNA was used to suppress the HMGB1 protein expression in the rat brain. The effect of RNA interference was evaluated by the methods of double-immunofluorescence labeling of HMGB1/GFAP and Western blotting. The proliferation of neural stem cells in the peri-infarction cortex was assessed by double labeling of BrdU/nestin. RESULTS：The protein expression of HMGB1 in I/R group was much higher than those in sham group (P<0.05). RNA interference effectively inhibited the HMGB1 expression (P<0.05). Double labeled BrdU/nestin positive cells in I/R group were more than that in sham group (P<0.05). The double labeled BrdU/nestin positive cells were significantly decreased in RNA interference group (P<0.05). CONCLUSION：Focal cerebral ischemia/reperfusion injury promotes the proliferation of neural stem cells in peri-infarction cortex by increasing HMGB1 protein level.     

Is That a Rodent in Your Luggage? A Mixed Method Approach to Describe Bushmeat Importation into the United States

Bushmeat, defined as meat derived from wild animals, is a potential source of zoonotic pathogens. Bushmeat from restricted animals is illegal to import into the United States under US federal regulations. We reviewed US Centers for Disease Control and Prevention (CDC) port of entry surveillance records from September 2005 through December 2010 and conducted focus group studies to describe trends in and reasons for bushmeat importation into the United States. In total, 543 confiscated bushmeat items were recorded. Half of the confiscated bushmeat items identified were rodents. Africa was the most frequent continent of origin. Seasonality was evident, with bushmeat confiscations peaking in late spring to early summer. Four times more bushmeat was confiscated during an enhanced surveillance period in June 2010 compared with the same period in previous years, suggesting that routine surveillance underestimated the amount of bushmeat detected at US Ports of Entry. Focus groups held in three major US cities revealed that bushmeat importation is a multifaceted issue. Longstanding cultural practices of hunting and eating bushmeat make it difficult for consumers to acknowledge potential health and ecologic risks. Also, US merchants selling African goods, including bushmeat, in their stores have caused confusion among importers as to whether importation is truly illegal. Enhancing routine surveillance for bushmeat and consistent enforcement of penalties at all ports of entry, along with health education aimed at bushmeat importers, might be useful to deter illegal importation.     

Lethal effects caused by suspended particles and sediment load on zoospores and gametophytes of the brown alga <Emphasis Type="Italic">Eisenia bicyclis</Emphasis>

ABSTRACT:   The effects caused by suspended particles in seawater and sediment cover of the substrate on the attachment to the substrate of zoospores, and on the subsequent growth and survival of gametophytes of the brown alga Eisenia bicyclis were examined in the laboratory. The attachment rate to the substrate of the zoospores was remarkably reduced by the sediment cover on the substrate, and it decreased to 3.8% of the no-sediment control by a slight sediment cover of 3.0 mg/cm² (0.048 mm in thickness). The growth of gametophytes was not inhibited at 10 mg/cm² or less of sediment cover, though it was stopped at 30 mg/cm². The survival rate of gametophytes became 39.4% at 5 mg/cm² on the substrate, and gametophytes were all blighted at 30 mg/cm². The ratio of male to female of the surviving gametophytes became 81.5:18.5, when the sediment cover was 10 mg/cm². The rate of total loss ( TL [%]) of zoospores and gametophytes of Eisenia bicyclis was obtained from the following equation: TL  = 100 (1 − exp[−0.0339  C ] exp[−1.24  Q ]), where, C (mg/L) and Q (mg/cm²) indicate the concentration of suspended particles in seawater and the sediment on the substrate, respectively.