猪流行性腹泻病毒S1D蛋白的优化表达及其单克隆抗体的制备

为进一步探究猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)S蛋白的抗原表位及其功能,本试验通过优化S1D基因的密码子,构建了S1D基因未优化的重组原核表达质粒pET-S1D和已优化的pET-ΔS1D,并进行了诱导表达和纯化。使用SDS-PAGE和Western blotting方法验证S1D、ΔS1D蛋白在大肠杆菌内得到正确表达,利用Image J软件对S1D、ΔS1D蛋白表达量进行灰度扫描,通过t检验分析两者差异性。将纯化的ΔS1D 蛋白免疫 BALB/c小鼠,通过细胞融合、筛选及亚克隆,获得单克隆细胞株。利用体内诱生法制备抗PEDV S1D蛋白的单克隆抗体腹水,使用ELISA、Western blotting、间接免疫荧光试验3种方法对腹水效价及特异性进行检测和验证。SDS-PAGE和Western blotting结果显示,表达S1D、ΔS1D蛋白的样品均在34 ku处出现正确的目的条带。t检验结果表明, S1D、ΔS1D两者蛋白表达量差异极显著(P<0.01)。ELISA结果显示, 腹水的抗体效价达到了1∶1 000 000,腹水与PEDV病毒粒子和纯化后的ΔS1D蛋白反应均呈阳性,与PEDV N蛋白、pET-32a(+)空载体蛋白和猪繁殖与呼吸综合征病毒(Porcine reproductive and respiratory syndrome virus,PRRSV)、猪传染性胃肠炎病毒(Transmissible gastroenteritis virus,TGEV)、猪瘟病毒(Classical swine fever virus,CSFV)、猪德尔塔冠状病毒(Porcine deltacoronavirus,PDCoV)和猪急性腹泻综合征冠状病毒(Swine acute diarrhea syndrome coronavirus,SADS-CoV) 5种病毒反应均呈阴性。Western blotting结果显示,腹水与ΔS1D蛋白和PEDV S蛋白分别在34和180 ku处有特异性条带出现,与pET-32a(+)空载体蛋白、正常Vero细胞蛋白均无特异性条带出现。间接免疫荧光试验结果显示,腹水及阳性对照组均能使细胞出现特异性绿色荧光信号,而空白及阴性对照组均未见绿色荧光信号。密码子优化可在原核表达系统中显著提高重组蛋白的表达水平,本研究基于高效表达的ΔS1D蛋白,成功制备了1株能稳定分泌与 PEDV S蛋白特异性结合的单克隆抗体的细胞株,为进一步探究 PEDV S蛋白抗原表位及蛋白功能的研究奠定了基础。     

2020年新疆地区猪繁殖与呼吸综合征病毒抗体检测与分析

为了解2020年新疆地区猪繁殖与呼吸障碍综合征病毒（PRRSV）抗体水平及其消长规律,本试验采用间接酶联免疫吸附试验（ELISA）法对1 218份血清中PRRSV免疫抗体水平进行检测和分析。结果显示,PRRSV抗体平均阳性率为78.49%（956/1218）,高于国家规定标准（70%）。S/P的平均值为1.22±0.84,变异系数为69.45%。不同类别猪群抗体平均阳性率在59.19%~91.50%之间,有一定的差异。在调查的10个规模化养殖场中,9个场PRRSV免疫抗体阳性率达到国家标准。不同类别猪群PRRSV变异系数较高,需进一步对现有的免疫程序进行调整和完善,确保各猪群健康。     

2020年新疆地区规模化猪场猪PCV2抗体检测与分析

为了解新疆地区2020年规模化猪场猪圆环病毒2型的免疫抗体水平,选择新疆地区6个规模化猪场,针对不同日龄段的猪采集血清样品333份,采用猪圆环病毒2型ELISA抗体检测方法进行抗体检测分析。结果发现,2020年新疆地区规模化猪场猪圆环病毒2型平均抗体阳性率为77.78%（259/333）,平均抗体阳性率符合国家标准。但各猪场免疫效果参差不齐,抗体阳性率最高的为A场,阳性率达100.00%;阳性率最低的为C场,仅48.78%,低于（70%）的国家标准。不同日龄段的猪抗体水平也存在一定的差异,抗体阳性率最高为种公猪,达96.30%;而保育阶段猪抗体水平最低,仅有47.92%,低于国家标准。     

2019—2020年新疆部分地区猪PCV2的抗体水平检测及分析

为了解2019—2020年新疆地区猪圆环病毒病2型（PCV2）抗体水平及其消长规律,本试验采用间接酶联免疫吸附试验（ELISA）方法对475份血清中PCV2免疫抗体水平进行检测和分析。结果显示,PCV2抗体平均阳性率为69.68%（331/475）,未达到国家规定标准（70%）。S/P的平均值为1.56。不同类别猪群抗体平均阳性率在42.50%~86.67%之间,有一定的差异。在调查的5个规模化养殖场中,仅有2个场PCV2免疫抗体阳性率达到国家标准。各场应根据抗体检测结果,进一步对现有的免疫程序进行调整和完整,确保各猪群健康。     

寒冷地区草原有毒有害牧草监测与防治

草原是内蒙古地区草食畜牧养殖产业健康发展的基础,同时也是优质牧草的主要来源。有毒有害牧草生长扩散预示着草场退化,对当地草食畜牧养殖产业健康发展会造成毁灭性打击。有毒有害植物生长发育,不仅会危害牲畜健康且还会严重消耗土壤的水分和养分,妨碍优良牧草繁殖生长,使草场产量和质量显著下降。在长时间草场利用过程中,很多农牧民群众只注重索取,不注重生态环境保护,使草场生产能力和抵抗能力逐渐变差,给有毒有害牧草繁殖生长提供了条件。因此需要加强寒冷地区草原有毒有害牧草的监测,并采取针对性措施进行防控,降低发病率。该文主要探讨内蒙古呼伦贝尔市寒冷地区草原有毒有害牧草的种类,并提出相应的防治措施。     

Estimation of Runoff,Peak Discharge and Sediment Load at the Event Scale in a Medium‐Size Mediterranean Watershed Using the Annagnps Model

In this paper, the Annualized Agricultural Non‐Point Source (AnnAGNPS) model has been used to estimate runoff, peak discharge and sediment load at the event scale in a Mediterranean watershed. The study area is the Carapelle torrent, Southern Italy (area = 506 km²), where continuous rainfall, streamflow and sediment load data are available. Nineteen flood events have been registered in the period 2007–2009 and were used for the application of the model. The aim of the paper is to evaluate the predictive accuracy of the model at the event scale, in a medium‐size watershed, given the specific conditions of the semi‐arid environments. A sensitivity analysis has been carried out to assign the correct parameterization: the mean normalized output variation of the most meaningful input parameters pointed out the influence of the curve number on runoff, peak discharge and sediment load predictions (values greater than 1); the MN Manning's roughness coefficient and K, C and P factors of the universal soil loss equation showed a moderate influence on sediment load simulations (values between 0·5 and 1). The selection of the Soil Conservation Service synthetic storm types has been based on the observed storm events analysis to improve the peak discharge simulations. The model prediction has proved to be good for runoff (R² = 0·74, NSE = 0·75, W = 0·92) and peak discharge (R² = 0·85, NSE = 0·70, W = 0·94), and satisfactory for sediment yield (R² = 0·70, NSE = 0·63, W = 0·91). The relative error is lower for high events; this result is quite interesting in semi‐arid environments, where most of the annual sediment yield is concentrated in a few, severe events. Copyright © 2013 John Wiley & Sons, Ltd.     

插秧机作业面积测量远程监测系统软件设计——基于VisualBasic.NET 2010

为实现对插秧机作业区域自动识别和作业面积的自动测量,实时掌握跨区作业插秧机远程作业面积参量值,以VisualStudio平台上的VisualBasic.NET2010为开发环境,开发了一套高速插秧机跨区作业面积远程测量监测系统软件。该软件功能主要包括GPS定位轨迹及作业区域的识别、面积测量、数据通讯、数据显示和数据存贮。系统采用VisualBasic.NET2010语言进行开发,将远程跨区作业的插秧机面积测量监测数据经由GPRS网络进行无线传输到上位机,并采用SOCKET组件和ADO.NET技术实现系统数据通信,完成PC机与下位机之间数据的传输,对测量结果进行显示存储,同时通过系统误差分析完成系统的可行性判断。     

虚拟仪器在杏鲍菇生长环境监控系统中的应用

有机食用菌杏鲍菇的栽培是宁夏南部山区的特色产业,杏鲍菇的生物学特性直接决定了工厂化栽培的基本特征。温湿度、二氧化碳浓度等环境因子对杏鲍菇不同时期的生长有着重要影响,研究环境因子的调控理论和方法是提升杏鲍菇品质与产量的重要基础,因此需要对环境因子各种变量进行数据采集。为此,以三菱FX2NPLC、FX2N-4 AD模拟量输入模块,以及传感器搭建环境因子数据采集平台;并以Lab VIEW作为上位机,通过PC与PLC串口通信,实现对环境因子的实时监控及数据存储。该研究为后期分析温室内外环境因子与杏鲍菇生长发育状况的数据关系奠定了科学基础。     

基于无线传感器网络的水质监测系统研究

针对国内外水质环境研究中水质样品异地监测分析存在的巨大缺陷,研究了水质自动监测系统。文章介绍了基于无线通讯传感器网络的简易水质自动监测系统,实现水质数据自动采集和实时上传,可以实现对钱塘江源头水质（衢州水质）进行更好的监测及保护。     

Study on the Change Rule of Egg Yolk Antibody in Laying Hens after First Immunization by Japanese Encephalitis Virus

The purpose of this experiment was to study the immunization rule of the egg yolk antibody affected by different vaccines,immunization dose and injection ways and further to discuss the optimal immunization procedures of the laying hens for the preparation of egg yolk antibody against swine Japanese encephalitis virus.180 brown laying hens without any vaccines were selected and divided into 18 groups randomly,each group of 10 hens.Groups 1,2 were the control groups,injected with the sterile saline;Groups 3 to 10 were injected with subcutaneous or intramuscular injection,and the vaccine was injected with 0.2,0.5,1.0 and 1.5 mL successively.Groups 11 to 18 were also adopted two kinds of injection,followed by the same dose of vaccine immunization.Six eggs of each experimental group were gathered before immune day and after 3,7,10,14,18,21 and 28 days,the egg yolk antibody was extracted and the titer was determined.As a result,the egg yolk antibody titers of groups 1 to 6,11 and 12 were all 0,and no significant immune response produced;The hens from 7 to 10 groups were injected with the inactivated vaccine.After 7 days,the average antibody titer reached the peak,and the duration of the antibody was 14 days.The hens from 13 to 18 groups were injected with the attenuated virus vaccine.After 14 days,the average antibody titer reached the highest value,and the duration of the antibody was 21 days.The egg yolk antibody titers were not significantly different in the two compared experiment groups with the same injection dose but with different injection ways (P>0.05).With the same injection way of each experiment group,and the difference was significant (P>0.05).Compared with some groups with the same injection and vaccine,the titer of yolk antibody was gradually increased with the increase of the immune dose,and the difference was significant (P<0.05).The results showed that,no matter intramuscular or subcutaneous injection,in order to produce a significant immune response to hens,the immune antigen dose was 1.0 mL inactivated vaccine or 0.5 mL attenuated vaccine at least.Compared with the attenuated and inactivated vaccine,inactivated vaccine stimulated the body to produce the antibody faster,but the maintenance time was shorter;The lower dose of attenuated vaccine could stimulate the body to produce antibodies,but the speed was slower,the maintenance time was longer.