Histological observations of bovine tuberculosis in lung and lymph node tissues from British deer

Deer are recognized as hosts of Mycobacterium bovis and assessing the role of wild cervids in perpetuating tuberculosis among cattle has motivated extensive research on several continents. In this paper, the histopathology of lymph node and lung tuberculous granulomas in M. bovis positive British deer is presented. The overall aim was to seek further insights into the potential for onward transmission from infected deer to other species, including cattle. Samples were obtained from an extensive survey of wild mammals in South-West England and from statutory tuberculosis surveillance. M. bovis culture-positive samples were characterised microscopically as to their stage of lesion advancement, number of acid-fast bacilli and granuloma encapsulation. Seventy percent of the deer developed granulomas containing far greater numbers of M. bovis bacilli than typically reported in cattle. Red and fallow deer had the largest number of poorly encapsulated granulomas often containing many hundreds of bacilli. The results are consistent with infected wild British deer being a potential source of environmental contamination and onward transmission to other species. However, further work on levels of bacillary shedding is required before this can be confirmed.     

Protein characterization using electrophoresis and immunofixation; a case‐based review of dogs and cats

Protein electrophoresis and immunotyping can be a useful adjunct to the standard biochemical techniques for characterizing serum and urine proteins. This paper reviews currently available and commonly used methods for diagnostic protein electrophoresis, including both agarose gel and capillary zone electrophoretic techniques and total protein assessments. Immunofixation and immunosubtraction methods for identification of immunoglobulin location and class are also presented. Practical application of quality assurance and quality control strategies in compliance with American Society of Veterinary Clinical Pathology (ASVCP) best practices are discussed. Commonly encountered serum and urine electrophoretic diagnostic patterns, including electrophoretically normal, acute‐phase protein responses, polyclonal gammopathies, restricted polyclonal/oligoclonal gammopathies, paraproteinemias (monoclonal or biclonal gammopathies), and Bence‐Jones proteinurias are also reviewed using relevant case material. Cases in which immunofixation electrophoresis are particularly useful are highlighted, and methodologies to more accurately quantify serum monoclonal proteins (M‐proteins), monitoring tests commonly used in human medicine, are discussed.     

普通小麦-柔软滨麦草易位系M97抗条锈基因的遗传分析和分子作图

为了明确M97抗条锈性遗传规律,在苗期用7个小麦条锈菌系对M97与感病品种铭贤169的杂交后代F1、F2、F3和BC1代进行抗条锈性遗传分析,并对M97抗Sun11-4的抗条锈基因进行SSR分子标记。M97对Sun11-4和Sun11-11的抗病性均由1对显性基因控制,对CY29、CY30、CY33的抗病性由1显1隐2对基因共同控制,对CY31的抗病性由2对显性基因独立或重叠作用控制。以接种Sun11-4的F2代分离群体构建作图群体,筛选到Xwmc222、Xwmc147、Xbarc229和Xwmc339等4个与抗病基因连锁的SSR标记,其遗传距离分别为3.4、4.8、7.6和12.1 cM。将该抗病基因定位于小麦1DS染色体,且该基因不同于已知的抗条锈基因,暂命名为YrM97。用YrM97两侧遗传距离最近的2个标记Xwmc222和Xwmc147对42个黄淮麦区主栽小麦品种进行分子检测,仅有9.5%的品种具有与YrM97相同的标记位点。     

微生物源农药申嗪霉素的研制与应用

申嗪霉素是中国自主研发的一种新型微生物源农药,具有高效、安全、广谱等特点,其主要成分是甜瓜根际促生菌M18产生的次级代谢产物吩嗪-1-羧酸。文章重点就申嗪霉素产生菌M18的分离及其代谢产物的鉴定、申嗪霉素生物合成及调控机理最新研究进展、申嗪霉素高产工程菌株的构建和产业化、以及申嗪霉素大田防病试验结果及推广应用情况等进行详细综述,并对其抗菌作用机理进行探讨,旨在为申嗪霉素的生物合成机理研究、遗传和代谢改造以及推广应用提供参考。     

猪繁殖与呼吸综合征病毒GP5、M蛋白基因的克隆及其基因疫苗构建

参照GenBank中猪繁殖与呼吸综合征病毒(PRRSV)美洲型代表株VR2332 GP5和M蛋白基因序列,设计并合成2对引物,用RT-PCR方法分别扩增出PRRSV野毒株GP5和M蛋白基因603,525bp片段,并将其分别克隆到pMD18-T载体。测序正确后,将GP5和M蛋白基因分别克隆到真核表达载体pEGFP-C1上,成功构建基因疫苗表达载体pEGP5-C1和pEM-C1。小鼠免疫试验证实,这些基因疫苗质粒可以诱导小鼠产生特异性抗体,并在二免后1周开始检测到特异性淋巴细胞增殖反应。     

富硒大果山楂果实中有关酶活性及贮藏品质的研究

[目的]探索硒在大果山楂贮藏保鲜中的应用,并为开发富硒大果山楂提供理论依据。[方法]以富硒大果山楂为原料,对其果实中有关酶活性及贮藏品质进行研究。[结果]富硒大果山楂果实中过氧化物酶(POD)和多酚氧化酶(PPO)的活性比对照组分别降低了49.2%和37.5%,超氧化物歧化酶(SOD)和过氧化氢酶(CAT)的活性比对照组分别增加了16.7%和33.5%。大果山楂果实硬度为25.29 N/cm~2,维生素C的含量为1 186.50 mg/kg,可溶性总糖的含量为144.00 g/kg,可溶性固形物的含量为16.60%,较对照组分别增加了13.2%、9.5%、29.0%、3.7%。[结论]硒能延缓大果山楂果实的成熟衰老,改善其品质。     

禽流感病毒H9N2亚型三重PCR检测方法的建立

为建立简便快速检测禽流感病毒(avian influenza virus,AIV)并同时区分出H9、N2亚型的方法,本试验根据基因库中H9亚型AIV的HA基因、N2亚型AIV的NA基因及AIV的M基因序列,分别设计了3对针对这3种基因保守序列的引物,建立了AIV H9N2亚型的三重PCR检测方法。应用该方法对H9N2亚型AIV模板进行PCR扩增,可得到3条与试验设计相符的目的条带,分别为313 bp (HA基因)、451 bp (NA基因)和667 bp(M基因);对非H9亚型的N2亚型AIV模板进行扩增,出现2条特异性扩增条带,即451 bp (NA基因)和667 bp(M基因);对非H9、N2亚型AIV模板进行扩增则只出现一条目的条带,即667 bp(M基因);对其他禽呼吸道病原体进行PCR扩增,结果均为阴性。敏感性试验结果显示此三重PCR方法最低检出限为10^-2 ng/μL。应用所建立的三重PCR方法对120份临床病料进行检测的结果与病毒分离鉴定结果一致。各项试验结果均表明,该方法对于禽流感病毒尤其是H9、N2亚型禽流感病毒的检测具有快捷、特异、灵敏的特点。     

Vitreosity,its stability and relationship to protein content in durum wheat

High quality requirements are set on durum wheat (Triticum durum) from semolina mills and pasta producers. For the production of semolina and pasta with good cooking quality, high grain protein content and vitreosity is required. The dependency of vitreosity on protein content as well as its stability under the influence of humidity was not well investigated up to now. We (1) compared two methods to determine vitreosity, (2) investigated the relationship between vitreosity and protein content, (3) developed a method to analyze vitreosity under humidity, and (4) examined the relationship between protein content and agronomical as well as quality traits in durum wheat. The results showed that the formation of vitreous kernels greatly depends on the protein content. To evaluate the stability of vitreosity under the influence of humidity a new method was elaborated and employed to assess the durum germplasm under study. This revealed that vitreosity of a durum wheat variety depends on the potential to form vitreous kernels but also to maintain this vitreosity under the influence of humidity. Our results further show that protein content is a central trait in durum wheat that strongly influences important traits like grain yield, vitreosity, and b-value.     

H3N2犬流感病毒M1蛋白的原核表达及鉴定

为制备犬流感病毒(H3N2) M1蛋白纯品,针对M1基因序列设计引物,用聚合酶链式反应(PCR)扩增目的基因片段,扩增产物克隆至表达载体pET-SUMO中并转化至宿主菌BL21(DE3),诱导表达目的蛋白,探索纯化工艺,制备目的蛋白,并用Western blot检测纯化的M1目的蛋白。通过PCR成功扩增出大小为771 bp的M1基因,成功构建p ET-SUMO-M1表达载体,表达的融合蛋白相对分子量为41 kD,主要以可溶形式表达,纯化后获得蛋白纯品,Western blot检测显示用M1蛋白(28 k D)免疫小鼠制备的多抗能与制备的蛋白纯品发生特异性反应,从而证明蛋白纯品为M1目的蛋白。试验制备出的M1蛋白纯品可为进一步制备通用型抗犬流感病毒抗体提供纯品抗原。     

猪繁殖与呼吸综合征病毒GP3/GP5/M真核表达载体的构建及其体液免疫应答

本试验旨在构建表达猪繁殖与呼吸综合征病毒(PRRSV)GP3、GP5和M蛋白的真核重组质粒。以PRRSV LN株为模板,采用PCR方法扩增出GP3、GP5、M基因片段,将扩增的GP5、M通过Linker序列串联成GP5-M,然后将GP3与GP5-M双酶切后插入pcDNA3.1(+)构建重组质粒pcDNA3.1-GP3-GP5-M,将其转染COS7细胞。PCR鉴定表明重组质粒pcDNA3.1-GP3-GP5-M含有PRRSV GP3、GP5-M基因,间接免疫荧光检测表明GP3、GP5-M蛋白在COS7细胞内获得表达。Western blotting检测证实GP3、GP5、M蛋白获得正确表达,并且所表达的GP3、GP5、M蛋白是融合蛋白。将pcDNA3.1-GP3-GP5-M免疫BALB/c小鼠,首免后2周可检测到特异性PRRSV中和抗体,首免后8周中和抗体效价最高可达1∶32。进一步将pcDNA3.1-GP3-GP5-M免疫断奶仔猪,首免后4周即可产生1∶4～1∶8的中和抗体。本试验成功构建了表达PRRSV GP3、GP5和M融合蛋白的真核重组质粒pcDNA3.1-GP3-GP5-M,中和抗体检测表明pcDNA3.1-GP3-GP5-M具有良好的免疫原性,从而为PRRSV基因工程疫苗的研制奠定基础。