Protective Antigens from Virulent Isolates of Fusobacterium necrophorum FN(A)

通过胰蛋白酶裂解坏死梭杆菌FN(A)型毒力菌株菌体,分别以菌体裂解物上清和沉淀作为抗原免疫家兔制备血清。利用SDS-PAGE/Western-blot技术在菌体中筛选出4种具有免疫原性的组分,通过免疫后的攻毒试验,筛选出1种具有免疫保护性的抗原。抗原分离纯化后,进行N端氨基酸序列测定、免疫试验及生物学特性研究,据试验结果可初步判定该抗原为溶血素类似物或一种新发现的抗原物质。该抗原不仅能使机体产生保护性免疫反应,而且不同菌株中此种抗原间可产生明显的交叉免疫。A virulent isolate of the bacterium FN(A) was lysed with trypsin.Supernatants and precipitates of the bacterial lysates were used to prepare anti-FN serum by immunizing rabbits.Four immunogenic components were isolated using SDS-PAGE/Western-blot.Immunized rabbits were challenged,and one protective antigen was screened.After antigen isolation and purification,the N-terminal amino acid sequence was determined and an immunological test and hemolysis test were performed.Results suggested that this antigen is either a hemolysin analogue or a new antigenic substance.The antigen was able to induce a protective immune response as well as generate a marked cross-immune reaction among different bacterial strains.国家科技部奶牛专项(2002BA518A04);; 中国农业科学院特产研究所科研基金项目(tcs-20041

Establishment of a PCR Technique to Detection of I,II Groups of D.nodosus in Footrot in Ruminants

根据节瘤拟杆菌特有保守序列设计引物,利用聚合酶链式反应(PCR)建立了一种鉴别检测该菌的方法。对可能影响PCR检测结果的一系列因素进行了较详细的研究,选择了适宜PCR检测的快捷处理病样获得反应模板的方法,对PCR反应条件进行了优化,使对已知菌株培养物检测灵敏性最高可到30个菌/30μL反应体系;用广泛的相关菌株和菌群验证引物的特异性,证明设计的引物特异性强。将PCR方法用于临床病样的初步检测,部分病样PCR检测阳性。为用PCR方法检测节瘤拟杆菌引发的牛、羊、鹿腐蹄病奠定了基础。Detection assay by PCR(Polymerase chain reaction) was constructed to D.nodosus,that can discriminate two group of Dichelobacter nodosus:(1)Three primers were designed,based on the conserved region of aligned fimA sequences of groups I,II of D.nodosus,of which forward primers were identical to group I,II,and reverse primers was specific to each group I and group II respectively;(2)The optimal PCR conditions were determined,in 30μl reaction volume the primers detected at least 30 cells of D.nodosus in crude lysates;(3)A serial of PCR assay verified the primers was specific to D.nodosus but not to the other bacterial and cells.The developed PCR technique were evaluated with directly lysate of samples from lesions footrot,the results found a few sample were positive in PCR test.From the results,which is suggested the developed PCR technique is adapted to identify D.nodosus and discriminate its serogroups,It will become a practical method to diagnosis of footrot in ruminants.中国科技部奶业专项“奶牛蹄病高效疫苗的研究与产业化开发”(2002BA518A04

Initial Application of a PCR for Detection of D.nodosus

本研究在建立节瘤拟杆菌PCR检测方法的基础上,将PCR方法用于临床样品的检测。PCR检测发现制约PCR直接检测临床样品检出率的主要因素是临床样品中细菌外的抑制物;采自趾叉皮炎的临床样品16份PCR检测呈阳性,趾叉皮炎是腐蹄病的初期感染症状;而其它蹄病后期症状临床样品PCR检测均为阴性,表明PC R检测方法适宜腐蹄病的早期诊断。The developed PCR technique were evaluated with directly lysate of samples from lesions in footrot,the results found PCR test was accurate and successful corresponding with DNA sequencing and microscope observation to sample from interdigital dermatitis in footrot;which is early syndrome of footrot in ruminant,so the PCR test was practised to detect those sample from early lesion.From the results,it will become a practical method to diagnosis of footrot in an early infection.中国科技部奶业专项“奶牛蹄病高效疫苗的研究与产业化开发”(2002BA518A04)研究课题资

1990-2011年山东省旅游节庆的时空变异特征及机理/Temporal-spatial variability of tourism festivals and its mechanism in Shandong Province during 1990-2011[J]

以山东省为研究区域,运用半变异函数、克里格空间插值等地统计学分析方法,分析了1990、1999、2005、2011年4个时间点上旅游节庆活动的时空分布规律、结构特征及变异性,并建立了拟合模型.结果表明:①旅游节庆数量空间不均衡显著,总体上呈现出“东高西低”的“三级阶梯状”分布格局；②空间结构特征明显,正负相关并存,并随滞后距离的增加,正相关性削弱,负相关性增强；③与1990年相比,2011年基台值增加了20.61倍,变程值由115.5 km增至335km,结构方差比则由69％下降至26％,随机性因素的影响减弱,而结构性因素的作用增强；④所有方向的分维数值呈下降趋势,其空间差异性增大,且全方向和东—西方向的差异性最大,其余方向相对均衡,其发展空间格局的板块状异向性分布特征明显；⑤经济发展水平、旅游供给能力、交通可达性是引起山东省旅游节庆差异演化规律的3个主要因素