鸭疫里氏杆菌鸭源致病性大肠埃希氏菌和鸭沙门菌的RAPD研究

以 4株不同血清型的鸭疫里氏杆菌、5株不同血清型的鸭源致病性大肠埃希氏菌和 5株同一血清型的鸭沙门菌为研究对象 ,分别提取基因组DNA ,利用随机扩增多态性DNA(RAPD)技术对其基因组DNA进行了分析。结果 ,从 2 0条随机引物中筛选出了 38条能在这 3种菌中有较好多态性的随机引物 ,8个有效引物共扩增出了 10 2个DNA片段 ,其中 14个菌株共有的谱带仅有 1条 ,显示多态性的片段有 10 1个 ,占 99.0 % ;对RA的 4个菌株扩增出的谱带数为 5 1条 ,共同片段有 12条 ,多态性片段 39条 ,占 76 .5 % ;对沙门菌的 5个菌株扩增出的条带数为 4 6条 ,共同片段为13条 ,多态性片段 33条 ,占 71.7% ;对大肠埃希氏菌的 5个菌株扩增出的条带数为 4 8条 ,共同片段 4条 ,多态性片段 4 4条 ,占 91.7%。在 8条引物中进一步筛选出 1条引物G12 ,其图谱中 5 35bp的条带为鸭疫里氏杆菌所特有 ,330bp的条带为沙门菌所特有 ,12 2 7bp的条带为大肠埃希氏菌所特有 ,表明G12可作为分子标记用来鉴别这 3种细菌。     

鸭沙门菌的随机扩增多态性DNA分析

以5株同一血清型的鸭沙门菌为研究对象,分别提取基因组DNA后,用20条随机引物对其进行RAPD分析。结果,20条随机引物中不能扩出任何条带的引物有2条,能扩出条带但扩增图谱中没有特异性条带出现的引物有10条,能扩出条带且扩增图谱具有多态性的引物有8条。对筛选到的具有多态性的8条引物进行分析,共扩增出46个DNA片段,片段大小为0.2～3.2kb,其中5个菌株共有的片段有13条,显示多态性的片段有33条。     

鸡白痢沙门菌基因组差异片段的筛选与分析

采用抑制性消减杂交技术(SSH)对鸡白痢沙门菌C79-13株与鸡伤寒沙门菌Sg9株进行了基因组差异片段分析。结果,从C79-13株中共检出13个特异性差异片段。经同源分析,这些序列可分为3类:噬菌体相关序列、质粒相关序列和已知功能序列。这些差异片段包含一些重要的沙门菌毒力相关基因,如编码大肠杆菌素I、paJ蛋白、尾突蛋白、切除酶的基因。结果表明,鸡白痢沙门菌C79-13株与鸡伤寒沙门菌Sg9株基因组间存在较多差异基因,这些差异片段为确定鸡白痢沙门菌特异性遗传标志,建立分子鉴别新体系提供了基础。     

鸡白痢沙门菌S06004ΔspiC突变株的构建与鉴定

为了研究spiC基因对鸡白痢沙门菌致病性的影响,采用同源重组方法构建该基因缺失株。以鸡白痢沙门菌S06004基因组DNA为模板,PCR扩增spiC基因上、下游DNA片段作为等位基因;将上游片段、卡那霉素抗性基因(KmR)及下游片段依次连接并克隆到pGMB151自杀性质粒上,构建重组自杀质粒pGMB151-ΔspiC/KmR,通过大肠杆菌χ7213与鸡白痢沙门菌S06004固相杂交,将质粒转入S06004中,运用反向筛选法获得含KmR基因的spiC基因缺失株(ΔspiC/KmR);再使用编码FLP重组酶的温度敏感型质粒pCP20敲除KmR,获得无抗性ΔspiC。PCR鉴定和测序结果显示,spiC基因被成功缺失。血清型、生化特性、生长特性、耐药性等鉴定表明,S06004ΔspiC未发生理化特性改变。本研究成功构建了spiC基因缺失株S06004ΔspiC,为进一步研究鸡白痢沙门菌致病性变化及spiC基因的功能奠定了重要基础。     

应用复合PCR方法检测沙门菌的研究

参照文献报道的沙门菌Repeatse和hisJ基因片段的引物序列 ,设计并合成了 2对引物 ,对沙门菌属和非沙门菌属的标准菌株及分离菌株进行了复合PCR扩增反应。结果 ,沙门菌PCR产物均出现 199bp与 4 95bp的特异性DNA扩增条带 ,而非沙门菌均未出现扩增条带 ,证明这 2对引物具有沙门菌属特异性。将提取的沙门菌DNA做梯度稀释 ,测定该复合PCR体系的敏感度。结果表明 ,此体系可检出 10 3CFU/mL菌液提取的DNA模板。应用上述方法 ,对进境鱼粉阳性样品进行了检测 ,均出现阳性结果。本研究表明 ,复合PCR是一种特异、敏感、快速的沙门菌检测方法 ,可应用于口岸系统鱼粉、肉骨粉的日常检测。     

福氏志贺菌2a多药耐药基因marA的克隆及其原核表达

根据福氏志贺菌2a多药耐药基因marA的序列,设计并合成了1对特异性引物,在引物的5'和3'端分别加入含有BamHⅠ和SalⅠ限制性酶切位点的序列,以福氏志贺菌2a菌株基因组DNA为模板扩增了marA基因序列.将PCR产物与pMD18-T载体连接,转化到大肠杆菌DH5α.鉴定成功获得目的片段后,经BamHⅠ+ SalⅠ双酶切并与载体pET-30a连接,构建原核表达质粒pET-30a-marA,并将其转化宿主菌BL21(DE3)pLys,用IPTG诱导其表达.SDS-PAGE分析表明,重组载体pET-30a-marA可成功地在大肠杆菌中表达MarA蛋白.Western-blotting检测证明,该蛋白能与志贺菌阳性血清发生特异性反应.     

鸡白痢沙门菌等位基因特异性PCR检测方法的建立

根据鸡白痢沙门菌与鸡伤寒沙门菌的rfbS基因在第237和第598位碱基的不同,设计和合成了等位基因特异性PCR引物,建立了快速检测鸡白痢沙门菌的PCR方法,并应用该方法对鸡白痢沙门菌临床分离样品进行了PCR鉴定。结果显示,该PCR方法能够特异性地鉴定鸡白痢沙门菌,检测灵敏度达100 pgDNA。对35个经常规方法鉴定的鸡白痢沙门菌分离株应用等位基因特异性PCR方法进行鉴定,鉴定出33株鸡白痢沙门菌,符合率为94.3%。表明,建立的等位基因特异性PCR方法能够准确而快速地鉴定鸡白痢沙门菌。     

应用PCR-RFLP分析鉴别广西鸡毒霉形体

针对鸡毒霉形体 (MG) 16S和 2 3SrRNA基因间的高变区序列设计合成了 1对引物 ,对MG菌株进行了PCR扩增 ,获得了 1条 899bp的DNA片段 ,而其他禽病病原DNA无扩增。扩增产物经用限制性内切酶AfaⅠ和DraⅠ进行酶切片段长度多态性分析 ,表明各MG菌株酶切图谱间均存在差异。     

大肠埃希氏菌和沙门菌氨基糖苷耐药基因PCR检测方法的建立

采用平板稀释法,选用氨基糖苷类药物链霉素、庆大霉素、卡那霉素和新霉素对22 株猪、鸡大肠埃希氏菌和30株猪、鸡沙门菌进行了药敏试验。结果显示,在4种药物中大肠埃希氏菌对卡那霉素的耐药率最高,达81.8%,而沙门菌则对链霉素的耐药率最高,达60%,两类细菌都对新霉素的耐药率最低。设计了5对引物,对耐药基因aph(3′) Ⅱ、aadA1、aadA2、aadB和aac(3) Ⅰa进行了扩增,获得的基因片段与预期的大小一致;序列分析表明,扩增产物与GenBank中的相应序列有很高的同源性(>99.8%)。采用建立的PCR技术,对22株猪、鸡大肠埃希氏菌和30 株猪、鸡沙门菌的耐药基因进行了检测,证实耐药基因普遍存在,在大肠埃希氏菌中aadA1 基因的检出率最高(12/22),在沙门菌中aph(3′) Ⅱ基因的检出率最高(18/30),在所有的菌株中都未能检测出aac(3) Ⅰa基因。     

进口鱼粉和肉骨粉病原性沙门菌的分离鉴定及其16S rRNA序列的同源性分析

从进口的 17批饲料用鱼粉、肉骨粉样品中分离得到 4株细菌 ,采用生化和血清学方法鉴定 ,提取其DNA ,用细菌 16SrRNA基因通用引物进行PCR扩增 ,并对PCR扩增产物测序 ,将测定的 16SrRNA序列在NCBI数据库中进行序列同源性比较 ,确证为沙门菌 ,其中样品 2、8与鼠伤寒沙门菌的序列同源性大于 99% ,样品 10、11与伤寒沙门菌的序列同源性大于 99%。由此可以认定样品 2、8为鼠伤寒沙门菌 ,样品 10、11为伤寒沙门菌     

痢疾杆菌福氏2a及其环丙沙星耐药株gyrA和parC基因耐药性决定区序列的测定分析

测定了痢疾杆菌福氏 2a 30 1株与喹喏酮类药物耐药性相关的 gyrA基因和 parC基因的序列 ,并对其环丙沙星耐药诱变株的 gyrA和 parC基因喹喏酮类药物耐药性决定区 (QRDR)序列进行了测定分析。结果表明 ,痢疾杆菌福氏 2a 30 1株 gyrA和 parC基因分别为 2 6 2 5bp和2 2 5 6bp ,环丙沙星诱导的耐药菌 gyrA基因QRDR(2 4 5bp)发生氨基酸残基 6 9 Ala→Val和 87 Tyr→Asp改变 ,parC基因QRDR(2 37b)发生氨基酸残基 79 Ala→Asp、84 Ala→Glu和 85 Pro→Ala改变。这一研究结果对认识痢疾杆菌喹喏酮类药物耐药性的分子机理具有重要意义     

Population, Prophets, Pensions, and Politics

As immigrant populations in Western welfare states grow at a faster rate than the native populations, whose birthrates have declined dramatically in recent years, Europe's tradition of democracy and tolerance is threatened. The reasons for the birthrate decline and the resultant aging of the native population are many and complex, but one important contributing factor is the pay-as-you-go pension system, which reduces people's immediate dependence on children. Moreover, the payroll and social security taxes that support the welfare state reduce the earnings capacity of men and women of the traditional age for having children, thus pushing down the birthrate. It is time to consider whether modern Europe's small-size families and high pensions are sustainable     

Terrorism,counter-terrorism,and critique: opportunities,examples, and implications

This article explores the parameters, value and limitations of different critical strategies for those dissatisfied with the contemporary politics of terror. It argues, first, that the prominent (counter-)terrorism paradigm – in which terrorism is approached as a ubiquitous and very specific security challenge meriting appropriately exceptional responses – is far more critiqued than we might anticipate. And, second, that such critiques – which can be found across political language, popular culture, everyday life, and beyond – employ distinct critical resources to serve varying ends. Reflecting on these, the article offers a new heuristic distinguishing five critical strategies which seek to: (i) repudiate; (ii) question; (iii) subvert; (iv) replace, or (v) deconstruct the prominent counter-terrorism paradigm. This typology, it argues, offers scope for optimism and strategic resources for those attracted to a critical terrorism studies project going forward.     

Pakistan,India, China,and the United States: Energy,Climate Change,and National Security

This article focuses on the interplay of energy, climate change, and national security issues in Southwest Asia, using the newer definition of “national security” to include energy security, economic development, and climate change, as well as traditional security focusing on the military aspects.     

Norway,Spitsbergen, and America, 1905-1920

In the Spitsbergen treaty of 1920, Norway acquired sovereignty over the Spitsbergen Islands. Rather than Woodrow Wilson, the American president, the architects behind the treaty were Robert Lansing, Wilson’s secretary of state, and, behind the diplomatic scene, the mining investor, John M. Longyear. In 1906, Longyear established a mining company to exploit the coal deposits at Spitsbergen. He induced Congress, the State Department, and the White House to forge an American policy for the European Arctic, including the appointment of Lansing, an international lawyer, as a counsel in the State Department. Lansing was a leading expert on both international law and the lack of state authority at the terra nullius, Spitsbergen. In 1915, he became secretary of State and, at the Paris Peace Conference, decided American policy regarding the Spitsbergen question. This analysis shows how the outcome of the Spitsbergen question was a result of American mining interests, supplemented by Norwegian-American shared interests in conflict resolution based on international law.     

International relations in Malaysia: theories, history, memory, perception, and context

The article makes a preliminary survey of the teaching of internationalrelations (IR) in Malaysia. It starts by describing the originsof the field, and the emergence of an IR epistemic communityjoining both academia and government. This account is necessarilyderived from the experiences of the four most established Malaysianuniversities distinguished by length of existence and officialfavor. Subsequently, the survey would describe course contentand influences going into their design. The penultimate sectionswould attempt to place the evolution of Malaysian IR teachingwithin a historical context. This survey nonetheless concludesthat nationalist aspirations continue to remain a secondaryinfluence when compared with intellectual dependence upon theWest in the design of IR education in Malaysia. Received for publication August 28, 2008. Accepted for publication October 2, 2008.     

Indigeneity,Ethnicity, and the State: Australia,Fiji, and New Zealand

This article draws on the politics of indigeneity to distinguish the claims of first occupancy from simple ethnic identity politics, illustrating that relative political marginalization in Australasia is not so much a function of minority status but of indigeneity itself. The politics of indigeneity's aim is to create political space for self-determination and a particular indigenous share in the sovereign authority of the nation-state itself. The Australasian states are compared with Fiji to demonstrate that the significance of historical constraints on political authority transcend the withdrawal of a colonial power and the restoration of collective indigenous majority population status.     

Culture,liberation, and 'development'

Most of the socio-economic changes taking place in Africa and much of the South are externally driven. External agencies, often in league with the State, by-pass working people and do not involve them in the decision-making processes. Their economic approaches ignore people's cultures and their world view. This denies working people a creative capacity to adapt new techniques and knowledge to their own concrete reality. This article argues for the importance of the historical frame of reference and for the centrality of culture in socio-economic processes. The author argues against approaches which are not culturally familiar to working people.