A new Brucella canis species-specific PCR assay for the diagnosis of canine brucellosis

Brucellosis is a zoonotic disease that is transmitted from animals to humans, and the development of a rapid, accurate, and widely available identification method is essential for diagnosing this disease. In this study, we developed a new Brucella canis species-specific (BcSS) PCR assay and evaluated its specificity and sensitivity. A specific PCR primer set was designed based on the BCAN_B0548-0549 region in chromosome II of B. canis. The PCR detection for B. canis included amplification of a 300-bp product that is, not found on other Brucella species or, genetically or serologically related bacteria. The detection limit of BcSS-PCR assay was 6 pg/μl by DNA dilution, or 3 × 10³ colony-forming units (CFU) in the buffy coats separated from whole blood experimentally inoculated with B. canis. Using the buffy coat in this PCR assay resulted in approximately 100-times higher sensitivity for B. canis as compared to detect directly from whole blood. This is the first report of a species-specific PCR assay to detect B. canis, and the new assay will provide a valuable tool for the diagnosis of B. canis infection.     

The siderophore-interacting protein is involved in iron acquisition and virulence of Riemerella anatipestifer strain CH3

Riemerella anatipestifer causes epizootic infectious disease in poultry and serious economic losses especially to the duck industry. However, little is known regarding the molecular basis of its pathogenesis. The ability to acquire iron under low-iron conditions is related to the virulence of a variety of bacterial pathogens. In this study, a sip (Riean_1281) deletion mutant CH3Δsip was constructed and characterized for iron-limited growth, biofilm formation, and pathogenicity to ducklings. Results showed that siderophore-interacting protein (SIP) was involved in iron utilization and the sip deletion significantly reduced biofilm formation and adherence to and invasion of Vero cells. In addition, the sip gene was absent in 1 of 24 (4.17%) virulent strains and 2 of 3 (66.7%) avirulent strains of R. anatipestifer, and the sip gene from six R. anatipestifer strains, which belong to serotypes 1, 2, and 10, respectively, shared 100% amino acid identities to those of R. anatipestifer strains DSM15868 and RA-GD. These results suggested that siderophore-mediated iron acquisition may be an important iron-uptake pathway in R. anatipestifer. Animal experiments indicated that the median lethal dose of the CH3Δsip mutant in ducklings was about 35-fold higher than that of the wild-type CH3 strain. Thus, our results demonstrated that R. anatipestifer SIP was involved in iron acquisition and necessary for its optimal virulence.     

水牛RNA聚合酶Ⅲ启动子的克隆与鉴定

[目的]获得有效的水牛RNA聚合酶Ⅲ启动子序列,为开展水牛源细胞的基因特异沉默研究奠定基础.[方法]通过启动子上、下游保守序列对水牛源启动子7SK、U6进行克隆和启动子关键顺式作用元件识别,利用一段针对EGFP的shRNA片段（shEGFP）对水牛7SK、U6启动子进行功能性分析,然后分别在水牛源及鼠源细胞中与pEGFP-N1共转染,转染48h后用荧光显微镜检测EGFP的表达情况,并用流式细胞仪和荧光实时定量PCR检测EGFP沉默表达情况.[结果]克隆获得水牛7SK、U6启动子序列分别为430和357 bp,其OCT-1（或CACCC盒）及TATA盒高度保守.连接shEGFP后与pEGFP-N1共转染细胞,通过荧光显微镜可观察到细胞发生了明显的荧光表达沉默现象;通过流式细胞分析,发现水牛7SK和U6启动子引导的shEGFP在水牛源细胞中沉默效率高达93.82％和87.45％;荧光实时定量PCR检测结果显示,在转染bu7SK-shEGFP和buU6-shEGFP的水牛源BFF细胞中,EGFP表达水平均显著低于其他物种启动子的细胞转染组（P＜0.05）,而在鼠源PT67细胞中,水牛启动子的启动效率与其他物种启动子差异不显著（P＞0.05）.[结论]水牛7SK和U6启动子可高效启动shRNA表达,且具有一定的物种特异性.     

淀山湖鱼类群落结构多样性的年际变化

淀山湖是上海市内陆水域最大的淡水湖泊,主要的淡水渔业水域和水产品来源地,也是重要的水生生物保护基地。为了评估增殖放流和生态环境变化对淀山湖鱼类群落结构变化的影响,本研究以2010-2012年淀山湖的渔业调查资料为基础,对该湖泊的鱼类优势种组成和多样性指数进行年际变化分析,并应用丰度生物量曲线方法对该湖的鱼类群落状况进行分析。结果表明:2010-2012年淀山湖鱼种类数基本稳定,基本鱼种组成变化不显著,优势鱼种组成趋势渐以小型鱼类为主;群落结构多样性指数年间差异不显著(P0.05);ABC曲线显示2010-2012这三年淀山湖群落的数量优势度曲线均高于生物量的优势度曲线,鱼类群落结构仍处于严重干扰状态。因此,建议改善增殖放流鱼种和加强渔业管理,以维持淀山湖鱼类群落的稳定。     

Gene Expression in Black Tiger Prawns Following Intramuscular Injection of β-gal Plasmid

Gene expression in black tiger prawns (Penaeus monodon) was studied following intra-muscular injection of CMVGal plasmid into the second abdominal segment. We used an in situ staining technique to detect -gal expression in one- and three-month-old injected prawns. We found that only one of the three-month-old prawns expressed the marker gene (2 days after injection), and the site of expression was confined to the sixth abdominal segment away from the injection site. We repeated the experiment on a new batch of three-month-old prawns, but using fluorometric determination technique. This time we found that -gal expression was detected (6/42) at the site of injection after 2, 7, and 14 days. In two other test samples, transgene expression was detected in the sixth abdominal segment only, further confirming the possibility of injected DNA dispersal. The results of the study also suggest that direct gene transfer is a feasible technique in black tiger prawns.     

东风11 G型内燃机车牵引缓冲装置关键部件检修工艺

介绍了东风11 G型内燃机车牵引缓冲装置的作用及主要结构,并以其关键部件车钩、缓冲器为研究对象,研究其检修工艺。     

新生代员工企业文化内部整合感知对工作满意度的影响研究——基于汽车整车制造业的实证数据

企业文化内部整合两个特质“参与性”与“一致性”对于工作满意度存在积极影响。相比传统员工,新生代员工对于企业文化的内部整合感知对于工作满意度表现出更强的预测作用。管理者应注重新生代员工的参与性激励、团队凝聚力塑造,与其建立民主沟通模式。     

Ciguatoxin in moray eels raising the risk for seafood safety in Viet Nam

Fisheries Science - Recently, suspected ciguatera fish poisoning (CFP) cases caused by the consumption of moray eels, popular seafood for locals and tourists, have been reported in Viet Nam....     

奶牛分子育种专题学习网站的建设构想

阐述了建设奶牛分子育种专题学习网站的意义和构想,规划了网站的主要内容和发展方向,为相关专业的研究者和从业人员提供参考。