从腹泻金黄地鼠中分离出致病性大肠杆菌

从腹泻金黄地鼠中分离到 3株细菌 ,经染色镜检、生化反应鉴定和动物试验确证 :该菌为大肠杆菌。该试验为地鼠大肠杆菌的防制提供了流行病学资料与理论依据。     

16S rRNA在兽医病原菌分类鉴定中的应用

介绍16S rRNA基因的特点，阐述其用于病原菌分类鉴定的基本原理，综述其在兽医病原菌分类鉴定中的应用。     

Application of acoustic Doppler current profiler combined with a scientific echo sounder for krill Euphausia pacifica density estimation

ABSTRACT:   An application of the acoustic Doppler current profiler (ADCP, 153.6 kHz) in combination with a scientific echo sounder (EK60, 38 and 200 kHz) was investigated to estimate the density of krill Euphausia pacifica . The acoustic backscattering strength from sound scattering layers was compared with biomass estimates from midwater trawls. Euphausia pacifica was targeted among mixed species populations in the sound scattering layer in the offshore Funka Bay area of Hokkaido, Japan. The frequency characteristics of acoustic backscattering by krill were calculated using a distorted wave Born approximation scattering model at three frequencies. Krill aggregations identified from the EK60 data were extracted as the mean volume backscattering strength difference between two frequencies. They were then used to identify similar aggregations in the ADCP data by matching observation times and depths for the two methods, which were applied simultaneously. Results from the comparison of the mean volume backscattering strength and the density calculated from the ADCP and EK60 showed that ADCP can be used to measure density and spatial–temporal distribution of krill aggregations. Current speed and direction at the study site were found to be 16.1 cm/s and 187.0°, respectively, and krill speed and direction (including the current component) were found to be 19.8 cm/s and 172.2°, respectively. Based on the ADCP data, the net speed and direction of the krill aggregations were found to be 5.9 cm/s and 128.0°, respectively.     

基于系统辩识的PID参数自整定

This paper presented a metnod of self-determining PID parameters by using system idenification and gave a engineering sample.     

鹅细小病毒HG5/82株的分离鉴定及生物学特性的研究

本研究通过对1982年黑龙江某鹅场发生的鹅细小病毒病疑似病例的肝病变组织病毒分离,进行回顾性研究.将肝脏加PBS研磨后,-70℃/37℃反复冻融三次,除菌过滤后接种12日龄鹅胚.发现病毒对鹅胚的致死时间为5～6 d,胚体体表有轻微出血点.鹅胚尿囊液经磷钨酸染色,可见具有典型特征的细小病毒粒子.将该病毒尿囊液在12日龄鹅胚中连续传16代,随着传代次数的增加,胚体多集中在3～4 d死亡.病毒传至第三代时死亡胚体头、颈、背部出现较为严重的出血点,体表有水肿.第五代病毒尿囊液(命名为HG5/82)对12日龄鹅胚的ELD50为10-5.92/0.1 mL.将20只12日龄雏鹅分为三个试验组和一个阴性对照组,各组分别接种鹅胚尿囊液104.92ELDso、103.92ELD50、102.92ELD50及生理盐水,发现雏鹅接种HG5/82后4 d出现轻微腹泻,随后恢复正常.用套式PCR检测雏鹅肛拭子病毒的体外排放情况.发现三个试验组在接种病毒后1～20 d用套式PCR均可检测到病毒.对照组及试验组接种后20～56d没有检测到病毒.将套式PCR产物进行序列测定并于参考毒株进行比较,表明该序列具有GPV相应区域的共同分子特征,与GPV参考毒株B的相应序列同源性达93.6%.本研究结果表明,该分离毒株为典型的鹅细小病毒(Goose Parvovirus,GPV),对雏鹅的致病力较弱.     

酥梨采后灰霉病生防菌株的鉴定及抑菌作用

以微生物拮抗病原菌是防治病害安全高效的可行手段。通过对果实表面微生物进行筛选鉴定,以期得到一株可以有效抑制灰霉病的生物防治菌株。对拮抗菌株H-1进行形态学特征、生理生化特性分析,结合16S rDNA序列分析和gyrB 基因序列分析构建系统发育树;采用平板对峙法结合果实伤口试验考察拮抗菌H-1在体外及梨果表面的抑菌作用。结果表明：经鉴定H-I菌株为贝莱斯芽孢杆菌（Bacillus velezensis）;B. velezensis H-1在体外条件下可显著抑制灰葡萄孢霉（Botrytis cinerea）、细极链格孢（Alternaria tenuissima）和串珠镰孢菌（Fusarium verticillioides）的菌丝生长,其中对灰葡萄孢霉菌丝的抑制作用最强;果实伤口试验表明,1×1011 CFU/mL的B. velezensis H-1可完全抑制梨果表面灰霉菌生长。由此可见,B. velezensis H-1可以有效抑制梨果采后灰霉病的发生。     

Detection of squash mosaic virus in seeds of melon (Cucumis melo) by enzyme-linked immunosorbent assay (ELISA)

Squash mosaic virus (SqMV, comovirus) is seed-transmitted in severalCucurbitaceae. Therefore, the use of virus-free seed is important to prevent establishment of this virus in the Netherlands and to avoid spread to other countries.This study was undertaken to develop an enzyme-linked immunosorbent assay (ELISA) for the detection of SqMV in melon seeds. An antiserum was produced to a serotype 1 isolate from melon. Two ELISA variants were investigated viz. an ELISA variant with simultaneous incubation of sample and enzyme conjugate (ELISA 1) and an ELISA variant with successive incubation of sample and enzyme conjugate (ELISA 2). The sensitivity of ELISA was tested by mixing fluor of ground infected and non-infected seeds in different proportions. SqMV was detected by both ELISA variants at dilutions of 1 160 (1 part of infected flour mixed with 159 parts of non-infected flour) or higher after a substrate incubation period of 4 h. However, ELISA 1 gave relatively higher absorbance values than ELISA 2 for nearly all dilutions. Since ELISA 1 is also faster than ELISA 2, ELISA 1 is advised for routine testing. In these test, using subsamples of 100 melon seeds SqMV is detected reliably. ELISA 1 is now used in the Netherlands for routine-indexing of melon seed lots for SqMV.Samenvatting Het pompoenemozaïekvirus gaat over met het zaad van verscheideneCucurbitaceae. Het gebruik van virusvrij zaad is belangrijk om te voorkomen dat het virus zijn intrede doet in Nederland en zich naar andere landen verspreidt.Een antiserum werd geproduceerd tegen een serotype 1 isolaat van meloen. Met behulp van dit antiserum werd een ELISA ontwikkeld om pompoenemozaïekvirus in zaden van meloen aan te tonen. Twee varianten van ELISA werden vergeleken, namelijk een variant waarbij monster en enzymconjugaat gelijktijdig geïncubeerd werden (ELISA 1) en een variant waarbij monster en enzymconjugaat na elkaar geïncubeerd werden (ELISA 2). De gevoeligheid van de ELISA varianten werd uitgetoetst door meel van zieke zaden in verschillende verhoudingen te mengen met meel van gezonde zaden. Het pompoenemozaïekvirus werd met beide ELISA varianten aangetoond in verdunningen van 1 160 (1 deel meel van zieke zaden gemengd met 159 delen meel van gezonde zaden) of hoger na 4 uur incubatie met substraat. ELISA 1 gaf doorgaans hogere extinctiewaarden dan ELISA 2 voor bijna alle verdunningen. Omdat ELISA 1 ook nog sneller is dan ELISA 2, wordt ELISA 1 aanbevolen voor routinematig gebruik. Wanneer voor routinematig gebruik 100 meloenezaden per submonster getoetst worden, kan het pompoenemozaïek virus betrouwbaar worden aangetoond. In Nederland worden momenteel per zaadpartij 20 submonsters van 100 zaden getoetst.     

菊花18个品种的RAPD分析

 采用RAPD 技术分析了18 个菊花品种DNA 的多态性。从80 个10 碱基随机引物中筛选出多态性频率高的3 个引物。扩增的多态性片段在600 bp～1300 bp 之间。检测出两个品种特有的分子标记, ‘大红托桂’有OPD15 (1200 bp) ,‘玉翎管’缺失OPA17 (1100 bp) 。瓣型一致的品种间基因型相似系数较高。     

Characterization of phytoplasmas detected in Chinaberry trees with symptoms of leaf yellowing and decline in Bolivia

Polymerase chain reaction (PCR) assays were used to detect phytoplasmas in foliage samples from Chinaberry ( Melia azedarach ) trees displaying symptoms of yellowing, little leaf and dieback in Bolivia. A ribosomal coding nuclear DNA (rDNA) product (1·8 kb) was amplified from one or more samples from seven of 17 affected trees by PCR employing phytoplasma-universal rRNA primer pair P1/P7. When P1/P7 products were reamplified using nested rRNA primer pair R16F2n/R16R2, phytoplasmas were detected in at least one sample from 13 of 17 trees with symptoms. Restriction fragment length polymorphism (RFLP) analysis of P1/P7 products indicated that trees CbY1 and CbY17 harboured Mexican periwinkle virescence (16SrXIII)-group and X-disease (16SrIII)-group phytoplasmas, respectively. Identification of two different phytoplasma types was supported by reamplification of P1/P7 products by nested PCR employing X-disease-group-specific rRNA primer pair R16mF2/WXint or stolbur-group-related primer pair fSTOL/rSTOL. These assays selectively amplified rDNA products of 1656 and 579 bp from nine and five trees with symptoms, respectively, of which two trees were coinfected with both phytoplasma types. Phylogenetic analysis of 16S rDNA sequences revealed Chinaberry yellows phytoplasma strain CbY17 to be most similar to the chayote witches'-broom (ChWBIII-Ch10) agent, a previously classified 16SrIII-J subgroup phytoplasma. Strain CbY1 resembled the Mexican periwinkle virescence phytoplasma, a 16SrXIII-group member. The latter strain varied from all known phytoplasmas composing group 16SrXIII. On this basis, strain CbY1 was assigned to a new subgroup, 16SrXIII-C.     

引起糖甜菜细菌性叶斑病的萎蔫短小杆菌新致病变种

 1995年在内蒙古临河市新发现了糖甜菜细菌性叶斑病,从病斑所分离的10个细菌菌株经柯赫氏法则验证,均确系该病的病原菌。采用形态观察、表型特征和生理生化特性测定、数值分析、血清学反应、细胞化学成分分析、DNA G+C mol%和DNA-DNA同源性测定进行了鉴定,并与植物病原棒形细菌15个标准菌株进行了比较。该病原菌为革兰氏阳性细菌,不规则短杆状,有一根鞭毛、亚极生或侧生,结合其生理生化特性、细胞化学成分和DNA G+C mol%和DNA-DNA同源性测定结果,认为应属于短小杆菌属(Curtobacterium)的萎蔫短小杆菌(Cur. flaccumfaciens),数值分析也支持这一结论。此外,据血清学反应结果及其对短小杆菌属的其它植物寄主的致病情况,认为该病原菌应是萎蔫短小杆菌种下的一个新的致病变种,定名为Curtobacterium flaccumfaciens pv. beticola pv. nov. Chen et al.,2000(萎蔫短小杆菌糖甜菜致病变种)。