日本脑炎诊断技术研究进展

日本脑炎是一种严重危害养猪业以及人类健康的虫媒病,每年造成大量的经济损失。为了更好地控制与预防该病,及时的诊断尤为重要。用于日本脑炎诊断的技术主要有病毒分离与鉴定、血清学诊断技术及分子生物学技术等。由于每种诊断技术都有其优势和不足之处,所以应根据具体情况选择合适的诊断方法。论文就各种诊断技术的原理、特点进行概述,并对日本脑炎诊断技术的发展趋势进行展望。     

Pseudomonas fluorescens contamination of a feline packed red blood cell unit and studies of canine units

Background: While screening programs have reduced the risk of infectious disease transmission by donors in human and veterinary blood banking, bacterial contamination of blood products has emerged as a major complication in human medicine. Objectives: To describe a Pseudomonas fluorescens (Pf)‐contaminated feline packed RBC (pRBC) unit and experimentally investigate Pf‐contaminated canine pRBCs. Methods: Canine pRBCs were inoculated with Pf‐rich pRBCs from the sentinel feline unit and stored at 4°C or 20°C for 72 hours. Aliquots from the pRBCs were serially evaluated by microscopy, culture, and a eubacterial 16S rRNA real‐time PCR assay. Results: One Pf‐contaminated feline unit turned black after 22 days of storage and was removed from the blood bank; a source was not found, and no other contaminated units were identified. Canine pRBCs spiked with 5 or 25 μL of the sentinel unit became culture‐ and/or 16S PCR‐positive at ≥8 hours at 20°C and 48 hours at 4°C and developed a color change at ≥24 hours. Sensitivity studies indicated that without incubation, inoculation of ≥100 μL Pf‐rich pRBCs was necessary for a positive 16S PCR test result. Conclusions: P. fluorescens grows in stored pRBCs slowly at 4°C and rapidly at 20°C. Screening of blood products for color change, estimating bacterial concentration with microscopy, and 16S PCR testing are simple and fast ways to detect bacteria in stored blood. Aseptic collection, temperature‐controlled storage, and regular visual monitoring of stored units is recommended. Discolored units should not be transfused, but examined for bacterial contamination or other blood product quality problems.     

新疆地区规模化奶牛场牛支原体流行病学调查

为了调查新疆地区规模化奶牛场牛支原体的感染情况,采用牛支原体间接ELISA和特异性PCR检测牛血清中的牛支原体抗体及病料中牛支原体核酸,共检测9个地区15个规模化奶牛场437份血清,肺脏、关节液及鼻腔黏液44份。结果显示,血清抗体阳性率为76.43%（334/437）,其中脐带血抗体阳性率为40.00%（4/10）;病料阳性率为40.91%（18/44）。检测结果表明,新疆地区大部分奶牛场存在牛支原体感染,部分奶牛场发生牛支原体肺炎及关节炎病例,并存在垂直传播的风险。牛支原体感染可能成为危害新疆规模化奶牛场犊牛健康的主要疫病之一。     

控制高径比条件下崩岗土体的收缩开裂特性

崩岗土体的收缩开裂受到多种因素的影响。该研究为研究高径比对其影响,共设计10组高径比,通过定点拍照记录脱湿前与脱湿结束时的土体形态变化,结合数字图像处理技术进行定量分析,探讨在控制高径比条件下崩岗土体的收缩开裂规律。结果表明：1）崩岗4层土中,过渡层的裂隙性、径向收缩性能最强,砂土层最弱,两者之间的较大差异会严重破坏崩岗土体的稳定性与承载力,促使崩壁崩塌;2）高径比较小的试样裂隙发育明显,径向收缩不明显;高径比较大的试样无裂隙发育,径向收缩显著。其中,4层土由干缩开裂土样过渡至径向收缩土样的高径比具体临界值分别位于：0.147～0.160、0.160～0.183、0.160～0.183、0.134～0.147;3）当高径比相同时,即使高度、直径不一致,但其各裂隙参数、径向收缩率具备相似性,轴向收缩率随厚度的增加而增加;4)随高径比的增加,收缩含水率逐渐增大,开裂含水率逐渐减小,两者之间的差值可以表示脱湿过程中土体产生抗拉强度的大小。收缩开裂裂隙度、宽径比、径向收缩率随高径比的增加整体呈现增大的趋势,其余参数均呈现减小的趋势。其中,4层土中,过渡层的收缩开裂特性受高径比影响最显著,砂土层受影响程度最小。研究结果可为揭示崩岗崩塌机理提供科学依据。     

Establishment of a Duplex Real-time RT-PCR Assay for the Differential Detection of Nipha Virus and Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus

To establish a rapid,sensitive and specific assay for the differential detection of Nipah virus (NiV) and highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV),a duplex Real-time RT-PCR was developed with specific primers and probes targeting to the special sequences of NiV M gene and HP-PRRSV nsp2 gene by optimization of reaction conditions.The performance of the assay was linear ranging from 4.6×10¹ to 4.6×10⁷ copies/μL for RNA standard control of NiV M (NiV-M-RNA) and from 4.1×10¹ to 4.1×10⁸ copies/μL for RNA standard control of HP-PRRSV nsp2 (HP-PRRSV-nsp2-RNA),and detection limits of the assay was 46 copies for the NiV-M-RNA and 4.1 copies for the HP-PRRSV-nsp2-RNA,respectively.The coefficients of variation (CVs) of both inter-assay and intra-assay repeatability were less than 2.0%,showing good repeatability.The assay was able to specifically detect NiV and HP-PRRSV simultaneously without cross-reaction with classical swine fever virus (CSFV),porcine epidemic diarrhea virus (PEDV),swine influenza virus (SIV),porcine parvovirus (PPV),pseudorabies virus (PRV) and porcine circovirus type 2 (PCV2).Of the 236 samples from pigs for both NiV and HP-PRRSV detection by the established assay,all the samples were negative for NiV,8 samples were HP-PRRSV positive.In conclusion,this assay offers a useful approach for the differential detection of NiV and HP-PRRSV in clinical specimens from the pigs.     

Test Analysis of Ecosystem on Functional Groups of Microorganism in Spiral Up- flow Reactor System

The test of functional groups of microorganisms in Spiral Up-flow Reactor System and the correlation between microorganisms in the water and the main environmental factors are studied.The results show that the diversity of microorganisms population in the SUFR system is complex and the bio-community formed in the SUFR system is stable.The close relations between abundance of bacteria and nutrients are found.The correlation between heterotrophic bacteria and COD is 0.949.The correlation between organic phosphate bacteria and TP is 0.815. The correlation between nitrosobacteria and NH_3-N is 0.909. The correlation between disnitrifier bacteria and TN is 0.653.     

辽西地区引种野杏数量性状研究

以辽西地区26个野杏无性系为试材,采用变异性分析、方差分析以及主成分分析等方法,研究了其25个数量性状的变异特点,以期为野杏无性系的遗传多样性研究和良种选育提供重要参考依据。结果表明:19个数量性状呈正态分布;25个野杏无性系数量性状变异系数均值23%,其中22个数量性状的变异系数在10%以上,小枝长度变异系数最大,达64%,单果质量、单核质量、单仁质量的变异系数分别为24%、21%和18%,表明野杏无性系数量性状变异程度丰富,选择潜力较大。野杏无性系果实性状重复力普遍偏高,核仁主要性状重复力次之,树体主要数量性状重复力较二者偏低,均达到极显著水平;果核仁性状的重复力除单果质量、单核质量和仁厚分别为0.634、0.756和0.624外,其它指标皆在0.909以上,其中仁长重复力最高,达0.973。主成分分析结果表明,前6个主成分累积贡献率达83.50%,能够反映野杏无性系数量性状的大部分信息;其中第1主成分主要反映野杏果核仁等经济性状。     

菠萝酸性转化酶基因家族初步生物信息学及表达分析

酸性转化酶（acid invertase, AIN）在菠萝采后蔗糖降解过程中起着重要作用,基于菠萝全基因组数据库,预测菠萝AIN家族基因并进行生物信息学分析,解析其在采后菠萝不同贮藏温度下的表达变化情况,为阐明AIN基因在采后菠萝果实贮藏特性中的作用奠定基础。以水稻AIN家族基因为探针,在菠萝全基因组中鉴定到2个菠萝细胞壁酸性转化酶基因（cell wall acid invertase, CWIN）和2个液泡酸性转化酶基因（vacuolar acid invertase, VIN）,分别命名为AcCWIN1、AcCWIN2、AcVIN1、AcVIN2,设计编码区引物进行测序验证,并进行生物信息学分析。进化分析结果表明,AcCWIN1、AcCWIN2和AcVIN1、AcVIN2蛋白分别归于细胞壁酸性转化酶和液泡酸性转化酶2个进化支上,且均属于糖基水解酶家族GH32,基因结构、保守域和保守基序均一致。荧光定量分析结果表明,菠萝果肉中AcVIN1和AcVIN2在果实采后贮藏过程中表达量升高,且AcVIN1在发生黑心病的部位大量表达,而AcCWIN1和AcCWIN2在采后贮藏过程中表达量逐渐降低,且随着贮藏温度的升高其表达量降低,预示AcVIN1、AcVIN2较AcCWIN1、AcCWIN2在菠萝采后蔗糖降解和黑心病的发生方面发挥着更为重要的作用。     

Quantitative trait loci mapping for yield-related traits under low and high planting densities in maize (Zea mays)

Average maize yield per hectare has increased significantly because of the improvement in high-density tolerance, but little attention has been paid to the genetic mechanism of grain yield response to high planting density. Here, we used a population of 301 recombinant inbred lines (RILs) derived from the cross YE478 × 08–641 to detect quantitative trait loci (QTLs) for 16 yield-related traits under two planting densities (57,000 and 114,000 plants per ha) across four environments. These yield-related traits responded differently to high-density stress. A total of 110 QTLs were observed for these traits: 33 QTLs only under low planting density, 50 QTLs under high planting density and 27 QTLs across both densities. Only two major QTLs, qCD6 and qWKEL2-2, were identified across low- and high-density treatments. Seven environmentally stable QTLs were also observed containing qED6, qWKEL3, qRN3-3, qRN7-2, qRN9-2 and qRN10 across both densities, as well as qRN9-1 under low density. In addition, 16 and eight pairs of loci with epistasis interaction (EPI) were detected under low and high planting densities, respectively. Additionally, nine and 17 loci showed QTL × environment interaction (QEI) under low- and high-density conditions, respectively. These interactions are of lesser importance than the main QTL effects. We also observed 26 pleiotropic QTL clusters, and the hotspot region 3.08 concentrated nine QTLs, suggesting its great importance for maize yield. These findings suggested that multiple minor QTLs, loci with EPI and QEI, pleiotropy and the complex network of “crosstalk” among them for yield-related traits were greatly influenced by plant density, which increases our understanding of the genetic mechanism of yield-related traits for high-density tolerance.     

基于NIR定量模型快速预测发酵麸皮中还原糖和可溶性蛋白成分的含量

为建立一种能快速预测发酵麦麸还原性糖和可溶性蛋白含量的定量分析模型,本研究以发酵麦麸为样本,采用3,5-二硝基水杨酸比色法（3,5-dinitrosalicylic acid, DNS）和BCA蛋白浓度测定法分别测定样品的还原性糖和可溶性蛋白含量;利用近红外光谱技术（Near infrared spectrum instrument, NIR）结合偏最小二乘法（Partial least squares regression, PLS）,比较预处理方法、最佳波长及主成分因子的决定系数R²,建立发酵麦麸还原性糖和可溶性蛋白含量的NIR快速检测定量模型。结果表明：1）发酵麦麸还原性糖定量模型的预处理方法使用一阶导数 （First derivative, FD）+二阶导数 （Second derivative, SD）+标准正态变换 （Standard normal variate, SNV）,光谱范围为908~1 670 nm ,主因子数为7时,模型效果最优,其决定系数Rc²为0.904 8,校正均方根误差（Square error corrected, SEC）为1.576 1,相对分析误差（Relative percentage difference, RPD）为3.240 8;外部验证集决定系数Rp²为0.954 9且还原糖活性成分的验证集样本的测定值与NIR光谱预测值的P值为0.959 5>0.05。2）发酵麦麸可溶性蛋白定量模型的预处理方法使用一阶导数 （First derivative, FD）+二阶导数 （Second derivative, SD）+标准正态变换 （Standard normal variate, SNV）,光谱范围为908~1 670 nm ,主因子数为10时,模型效果最优,其决定系数Rc²为0.938 2,校正均方根误差（Square error corrected, SEC）为2.003,相对分析误差（Relative percentage difference, RPD）为4.021 9;外部验证集决定系数Rp²为0.994 4,且可溶性蛋白活性成分的验证集样本测定值与NIR光谱预测值的P值为0.901 9>0.05。综上,建立的NIR光谱定量模型稳定性和准确性较好,且预测准确度良好,可用于快速预测发酵麦麸样品的还原性糖和可溶性蛋白含量。