Identification of a conservative site in the African swine fever virus p54 protein and its preliminary application in a serological assay

BackgroundASF was first reported in Kenya in 1910 in 1921. In China, ASF spread to 31 provinces including Henan and Jiangsu within six months after it was first reported on August 3, 2018. The epidemic almost affected the whole China, causing direct economic losses of tens of billions of yuan. Cause great loss to our pig industry. As ELISA is cheap and easy to operate, OIE regards it as the preferred serological method for ASF detection. P54 protein has good antigenicity and is an ideal antigen for detection.ObjectiveTo identify a conservative site in the African swine fever virus (ASFV) p54 protein and perform a Cloth-enzyme-linked immunosorbent assay (ELISA) for detecting the ASFV antibody in order to reduce risks posed by using the live virus in diagnostic assays.MethodWe used bioinformatics methods to predict the antigen epitope of the ASFV p54 protein in combination with the antigenic index and artificially synthesized the predicted antigen epitope peptides. Using ASFV-positive serum and specific monoclonal antibodies (mAbs), we performed indirect ELISA and blocking ELISA to verify the immunological properties of the predicted epitope polypeptide.ResultsThe results of our prediction revealed that the possible antigen epitope regions were A23–29, A36–45, A72–94, A114–120, A124–130, and A137–150. The indirect ELISA showed that the peptides A23–29, A36–45, A72–94, A114–120, and A137–150 have good antigenicity. Moreover, the A36–45 polypeptide can react specifically with the mAb secreted by hybridoma cells, and its binding site contains a minimum number of essential amino acids in the sequence 37DIQFINPY44.ConclusionsOur study confirmed a conservative antigenic site in the ASFV p54 protein and its amino acid sequence. A competitive ELISA method for detecting ASFV antibodies was established based on recombinant p54 and matching mAb. Moreover, testing the protein sequence alignment verified that the method can theoretically detect antibodies produced by pigs affected by nearly all ASFVs worldwide.     

宁夏两次夏季典型暴雨过程成因对比分析

利用自动气象站站点实况和ECMWF ERA5的逐时再分析资料,通过环流分析和物理量诊断方法,对比2017年6月4日和2019年8月2日宁夏区域性暴雨强降水区域、环流背景、影响系统和物理量因子的共性和不同.结果表明:后者累积降水量和最大小时雨强均比前者大;2017年6月4日,宁夏处于东高西低的环流形势中,强降水出现在低空急流的左前方,低空切变线的右前方,即宁夏中北部,稳定维持的地面中尺度辐合线触发了局地强降水;2019年8月2日暴雨过程出现在宁夏南北两头,是两槽一脊环流背景下,由500hPa冷涡、冷槽和700hPa低槽、切变、低空急流共同造成的,是一次典型的强对流单体导致的暴雨过程.2019年8月2日短时强降水过程比湿、CAPE、ZH零度高度、-20度层高度数值较大;两次短时强降水过程沙氏指数为正;2017年6月4日LI为正值,2019年8月2日LI为负值;2017年6月4日850-500假相当位温差为负值,2019年8月2日为正值;二者200hpa 、700hPa为辐散,850hPa为辐合,后者200hPa辐散中心强度大于前者,而700hPa辐散中心强度和850hPa辐合中心强度反之。     

禽I型副粘病毒广西分离株F基因序列及毒力的研究

对源于鸡、鹅、鸽的 9株APMV_1广西分离株及我国常用的中发型疫苗毒株I系 (Mukteswar株 )的F基因N_端前段进行了RT_PCR扩增 ,产物进行核苷酸序列测定 ,用基因分析软件DNAStar进行分析并与已发表的其它参考毒株进行比较。结果发现 ,广西分离株在裂解位点的氨基酸组成和排列均符合强毒株的特征 ;根据F基因绘制的系谱树来看 ,广西鸡和鹅分离株都归属于基因型VII,证实近年来广西流行的基因型与国内其他地区的相同 ;并发现中发型疫苗毒株I系 (Mukteswar株 )也属于基因型VII。研究还对分离株进行了常规的毒力和致病性测定试验 ,并与根据F基因序列特征判定的结果相比较 ,结果发现其中一株鸽源和两株鹅源分离株根据常规方法判定的结果与毒株在临床上的致病性不相符 ,而根据F基因序列特征判定的结果与毒株在临床上的致病性相一致     

新城疫病毒融合蛋白裂解位点基因的分离克隆及在原核细胞中的表达

用ＳＰＦ鸡胚繁殖新城疫病毒（ＮＤＶ）Ｆ４６Ｅ９株（强毒）、ＬａＳｏｔａＥ４株（弱毒），对病毒进行纯化，抽提ＲＮＡ。用１对均为２７个碱基的引物进行ＲＴ－ＰＣＲ，扩增出了２个毒株的融合蛋白裂解位点（Ｆｃ）基因。将Ｆｃ基因定向克隆入ｐＵＣ１８的ＥｃｏＲＩ和ＳａｌⅠ位点之间，获得２个毒株Ｆｃ基因克隆ｐＵＣＦ４６Ｆｃ和ｐＵＣＬａＦｃ，用ＥｃｏＲＩ／ＳａｌⅠ双酶切法、ＰｓｔⅠ单酶切法、ＰＣＲ法和核酸探针法对其进行了鉴定。将鉴定好的ＬａＳｏｔａＥ４Ｆｃ基因定向导入表达载体质粒ｐＢＶ２２１，获得重组子ｐＢＶＬａＦｃ。ＰＣＲ和内切酶酶切法鉴定表明，Ｆｃ插入的位置和方向都正确无误。用Ｅ．ｃｏｌｉＤＨ５α通过热诱导法进行了表达。用ＳＤＳ－ＰＡＧＥ和Ｗｅｓｔｅｒｎ－ｂｌｏｔ法检测了表达产物。结果表明，有１条能够与ＮＤＶ多抗反应的特异条带，分子量约１５７００，与预期大小一致，说明是Ｆｃ基因的表达产物     

2000-2009年新疆雷电灾情分析

对2000-2009年气象部门收集的雷灾资料进行统计分析,研究新疆雷电灾情的时空分布特征、受损财物与受伤害人员情况等。资料包括97次雷灾事件,其中30次雷灾造成了31人死亡,15人受伤。新疆雷灾发生在3~9月,夏季占78%以上,6月最高。造成人员伤亡和财产损失的雷灾主要发生在11:00~24:00时,17:00~18:00时为峰值。雷灾致人员伤亡的雷击地点,发生在户外的最多。     

Spatial Clustering of Escherichia coli with Reduced Susceptibility to Cefotaxime and Ciprofloxacin among Dairy Cattle Farms Relative to European Starling Night Roosts

European starlings (Sturnus vulgaris) have been implicated in the dispersal of zoonotic enteric pathogens. However, their role in disseminating antimicrobial‐resistant organisms through their home range has not been clearly established. The aim of this study was to determine whether starling night roosts served as foci for spreading organisms with reduced susceptibility to antimicrobials among dairy cattle farms. Bovine faecal pats were collected from 150 dairy farms in Ohio. Each farm was visited twice (in summer and fall) between 2007 and 2009. A total of 1490 samples (10 samples/farm over two visits) were tested for Escherichia coli with reduced susceptibility to cefotaxime and ciprofloxacin. Using a spatial scan statistic, focal scans were conducted to determine whether clusters of farms with a high prevalence of organisms with reduced susceptibility to cefotaxime and ciprofloxacin surrounded starling night roosts. Faecal pats 13.42% and 13.56% of samples carried Escherichia coli with reduced susceptibility to cefotaxime and ciprofloxacin, respectively. Statistically significant (P < 0.05) spatial clusters of faecal pats with high prevalence of Escherichia coli showing reduced susceptibility to cefotaxime and ciprofloxacin were identified around these night roosts. This finding suggests that the risk of carriage of organisms with reduced susceptibility to antimicrobials in cattle closer to starling night roosts was higher compared to cattle located on farms further from these sites. Starlings might have an important role in spreading antimicrobial‐resistant E. coli to livestock environments, thus posing a threat to animal and public health.     

北方农牧交错带土地利用及其生态背景分析

在遥感和地理信息系统技术支持下,提取年降水量、≥10℃积温、年平均风速、海拔高度、坡度等数据作为生态背景基本要素,基于NOAA/AVHRR影像分类而成的北方农牧交错带全样带1989年和2000年两期土地利用数据进行空间分析,分别研究单一和复合生态背景下土地利用的时空变化特征及其与生态背景的关系。结果表明:年降水量和年平均风速对土地利用变化的影响较为明显,特别是在坡度<15°的区域,降水对耕地和草地、年平均风速对林地和草地的影响均比较明显。≥10℃积温的变化与土地利用关系则不明显。耕地、林地、草地和城镇用地主要分布在海拔1 000~3 500 m和500 m以下、坡度为15°以下的地区,水域主要分布在海拔200 m以下、坡度为5°以下的地区。     

PCV2 Rep蛋白N-糖基化位点突变对病毒复制的影响

N-糖基化对病毒的感染与增殖均具有重要作用,与PCV2复制相关的Rep蛋白含有三个N-糖基化位点。为了分析PCV2Rep蛋白N-糖基化位点突变对病毒复制的影响,本试验构建了三个双拷贝突变体感染性克隆2M23、2M256、2M286,并成功拯救病毒。通过间接免疫荧光检测病毒的拯救效果,TCID50测定病毒的感染力,荧光定量PCR检测细胞病毒的载量。结果显示,PCV2Rep蛋白的23~25aa、256~258aa N-糖基化位点突变后降低病毒的复制能力,而286~288aa突变后增强病毒的复制能力,为进一步阐明PCV2的复制及致病机制提供参考。     

建筑电气现场施工问题与应对措施探讨

建筑电气现场施工是建筑工程项目重要的环节,关系到整个工程的质量。由于电气安装等问题引发的灾难和事故不在少数,对电气施工问题的处理方式影响着整个建筑工程。由此可见,加强对建筑电气现场施工环节的控制是非常重要的。文章主要就建筑电气现场施工问题及应对措施进行了探讨。     

A Gis-derived integrated moisture index to predict forest composition and productivity of Ohio forests (U.S.A.)

A geographic information system (GIS) approach was used in conjunction with forest-plot data to develop an integrated moisture index (IMI), which was then used to predict forest productivity (site index) and species composition for forests in Ohio. In this region, typical of eastern hardwoods across the Midwest and southern Appalachians, topographic aspect and position (rather than elevation) change drastically at the fine scale and strongly influence many ecological functions. Elevational contours, soil series mapping units, and plot locations were digitized for the Vinton Furnace Experimental Forest in southeastern Ohio and gridded to 7.5-m cells for GIS modeling. Several landscape features (a slope-aspect shading index, cumulative flow of water downslope, curvature of the landscape, and water-holding capacity of the soil) were used to create the IMI, which was then statistically analyzed with site-index values and composition data for plots. On the basis of IMI values for forest land harvested in the past 30 years, we estimated oak site index and the percentage composition of two major species groups in the region: oak (Quercus spp.), and yellow poplar (Liriodendron tulipifera) plus black cherry (Prunus serotina). The derived statistical relationships were then applied in the GIS to create maps of site index and composition, and verified with independent data. The maps show the oaks will dominate on dry, ridge top positions (i.e., low site index), while the yellow poplar and black cherry will predominate on mesic sites. Digital elevation models with coarser resolution (1:24K, 1:100K, 1:250K) also were tested in the same manner. We had generally good success for 1:24K, moderate success for 1:100K, but no success for 1:250K data. This simple and portable approach has the advantage of using readily available GIS information which is time-invariant and requires no fieldwork. The IMI can be used to better manage forest resources where moisture is limiting and to predict how the resource will change under various forms of ecosystem management.