检测青海省藏獒棘球绦虫虫卵方法的比较

应用蔗糖离心尼龙网筛法、福尔马林乙醚沉淀法和饱和蔗糖漂浮法,对来自青海省的83份藏獒的粪便,进行棘球绦虫虫卵检查。结果：上述三法的阳性检出率分别为18．07％（15／83）,15．66％（13／83）和13．25％（11／83）。     

应用荧光定量PCR对细粒棘球绦虫miRNAs在虫体不同发育阶段表达量的研究

为确定通过Solexa高通量测序鉴定出的大量细粒棘球绦虫miRNAs序列在不同生活史阶段的表达量。采用SYBR染料法和Race试剂盒相结合的荧光定量方法对原头节和生发层阶段的miRNAs表达进行了检测分析。结果表明：两阶段miRNAs表达差异较大,其中mir-71、mir-7、mir-1和mir-19在生发层阶段表达量较高,而let-7则在原头节阶段表达较高。说明这些miRNAs可能在细粒棘球绦虫发育阶段的基因转录调控中起着重要作用。     

用抗虫体表膜抗原抗体测定犬细粒棘球绦虫粪抗原

利用Echinococcus granulosus(Eg)成虫表膜抗原抗体夹心ELISA方法检测犬细粒棘球绦虫感染的粪抗原.首先提取Eg成虫表膜抗原,制备抗Eg成虫表膜抗原的血清,然后用间接ELISA和Western blotting方法检测其抗原抗体反应及免疫原性,用间接免疫组化方法对Eg成虫表膜蛋白进行组织定位.利...     

莫能菌素对细粒棘球蚴原头蚴作用的试验

将细粒棘球蚴原头蚴培养于含7μg/ml莫能菌素的NCTC135培养液中,对其在36小时内的活动及结构变化作了观察。光镜下观察,培养至12小时部份虫体停止活动;至24小时已有半数虫体结构模糊,美蓝着染证明部分死亡;至36小时所有原头蚴结构模糊,全部死亡。电镜下观察,虫体高尔基复合体最先出现退行性变化,随后线粒体结构破坏,进而引起整个胞质及胞核的改变,至36小时细胞死亡。文中就莫能菌素作用的特点及细胞器改变的意义作了讨论。     

细粒棘球绦虫脂肪酸去饱和酶1基因序列及不同发育阶段表达分析

旨在寻找细粒棘球绦虫发育相关基因,为预防和治疗细粒棘球绦虫引起的包虫病而制定相应的措施提供理论基础。通过分析细粒棘球绦虫原头蚴高通量转录组测序数据文库,对Unigene进行KEGG通路分析,筛选出一个脂肪代谢通路,并筛选出该通路中主要的调控基因脂肪酸去饱和酶1。通过对Eg-FADS1基因的PCR扩增,分析其核苷酸序列及氨基酸序列,用实时荧光定量PCR和全量组织原位杂交检测Eg-FADS1基因在不同发育阶段的表达情况。结果显示,该基因具有完整的开放阅读框,cDNA全长为819bp,编码272个氨基酸,预测该蛋白分子质量为30.98ku,等电点为7.10。实时荧光定量PCR结果表明,Eg-FADS1基因在原头蚴及成虫阶段均有表达,并且该基因在成虫阶段极显著高于原头蚴阶段的表达量。全量组织原位杂交结果显示,Eg-FADS1mRNA在原头蚴及成虫阶段分布广泛。提示Eg-FADS1具有生物防治作用,值得进一步研究。     

Molecular update on cystic echinococcosis in cattle and water buffaloes of southern Italy

Cystic echinococcosis (CE)--caused by the larval stage (hydatid cyst) of the cestode Echinococcus granulosus--is one of the most widespread zoonoses of veterinary and medical importance. Molecular techniques have allowed the identification of 10 different genotypes (G1-G10) of the parasite. The present paper is an update regarding the E. granulosus genotypes infecting water buffaloes and cattle bred in the Campania region of southern Italy. The molecular study was performed on 30 hydatid cysts (11 from water buffaloes and 19 from cattle). Two different mitochondrial DNA genes, namely the cytochrome c oxidase subunits 1 and the 12S ribosomal DNA (12S rDNA) were used as genetic markers. Three different genotypes of E. granulosus were unequivocally identified, i.e. the G1 (common sheep), G2 (Tasmanian sheep) and G3 (buffalo) genotypes, as well as some G1 and G2 variants. It should be noted that the present study demonstrated for the first time: (i) the presence of the G2 genotype in water buffaloes from a Mediterranean area; and (ii) the fact that the analysed portion of the 12S rDNA gene can not discriminate between the G2 and G3 genotypes of E. granulosus. The finding of the G1, G2 and G3 genotypes in large ruminants from southern Italy is of epidemiological relevance and immediate public health importance because of their recognized infectivity in humans.     

Echinococcus granulosus infection in Spain

Cystic echinococcosis (CE) caused by the cestode Echinococcus granulosus is an endemic disease in Spain. Although specific control programmes initiated in the 1980s have led to marked reductions in CE infection rates in Spain, the disease still remains an important human and animal health problem in many regions of the country. Human incidence and livestock (including sheep, cattle, pigs and horses) prevalence data were gathered from national epidemiological surveillance information systems and regional institutions for the period 2000-2005. Additionally, data on the prevalence of E. granulosus infection in dogs were obtained from published literature. The most affected regions were those of the North Eastern, Central and Western parts of the country, (Autonomous Regions of Aragon, Castile-La Mancha, Castile-Leon, Extremadura, Navarre and La Rioja), where human CE incidence rates in the range of 1.1-3.4 cases per 10(5) inhabitants coexist with ovine/bovine CE prevalence rates up to 23%. Control programmes of hydatidosis/echinococcosis should be reinforced in these regions to reduce the prevalence of the disease.     

细粒棘球绦虫miRNAs的鉴定及分析

细粒棘球绦虫具有复杂的基因调控模式,在中间宿主和终末宿主体内具有截然不同的生物学特性。miRNA是真核生物内存在的非常重要的调控因子,本研究采用Solexa高通量测序技术,对G1株原头蚴mi-croRNAs进行了鉴定和分析,总共发现了108条miRNAs,其中26条属于保守家族,82条是细粒棘球绦虫特有。     

细粒棘球绦虫BAG3和EB1基因的克隆、表达及其在原头蚴细胞凋亡中的作用

旨在探究细粒棘球绦虫BAG3(Eg-BAG3)和EB1(Eg-EB1)的分子特征及在原头蚴细胞凋亡过程中的作用,采用原核表达方法获得重组Eg-BAG3和Eg-EB1,利用生物信息学、蛋白免疫印迹及免疫荧光定位方法对Eg-BAG3和Eg-EB1的分子特征进行了初步探究,随后利用10 mmol·L^-1H₂O₂诱导原头蚴细胞凋亡,qRT-PCR方法检测Eg-BAG3和Eg-EB1的转录水平,并通过RNA干扰技术进一步探究二者与原头蚴细胞凋亡之间的关系。结果显示,Eg-BAG3含有BAG蛋白家族特征性的BAG结构域,Eg-EB1含有内吞蛋白家族特征性的BAR结构域,二者分别属于典型的BAG蛋白和内吞蛋白。免疫荧光定位结果显示Eg-BAG3广泛分布于细粒棘球绦虫的各个发育时期,Eg-EB1主要定位于原头蚴皮层、吸盘及顶突和包囊生发层,而在成虫未见特异性分布。H₂O₂可以成功诱导原头蚴细胞凋亡,且原头蚴中Eg-BAG3和Eg-EB1的相对转录水平都随着H₂O₂处理时间的延长而显著上升,在8 h后的相对转录水平与0 h相比具有显著性差异(P<0.05)。RNA干扰结果显示,Eg-BAG3干扰组原头蚴细胞凋亡率显著高于未干扰组(P<0.05),而Eg-EB1干扰组原头蚴细胞凋亡率低于未干扰组(P<0.05),表明Eg-BAG3在H₂O₂诱导的原头蚴细胞凋亡过程中起到抗凋亡作用,而Eg-EB1起到促凋亡作用。Eg-BAG3可以抑制氧化应激诱导的原头蚴细胞凋亡而Eg-EB1促进细胞凋亡,且二者可能参与调控不育囊的形成。