用ELISA法观察模型小鼠原发性感染细粒棘球绦虫后特异性抗体的变化规律

分别用较高纯度的细粒棘球绦虫六钩蚴抗原和传统使用的囊液抗原对一次感染和二次感染细粒棘球绦虫六钩蚴的小鼠血清进行ELISA测定，认为寄生于不同部位的包囊刺激机体产生的抗体水平不同．同时检测早期抗体时，六钩蚴抗原比囊液抗原更具有阶段性优越性。小鼠二次感染六钩蚴后六钩蚴抗体水平得到明显加强，并推测认为一次感染产生的六钩蚴抗体可能是具有保护性的抗体。     

新疆棘球蚴Eg95基因的克隆与序列分析

从新疆地区细粒棘球蚴原头节中提取基因组DNA,以细粒棘球蚴原头节DNA为模板,用Eg95特异性引物进行PCR扩增,获得Eg95-XJ-1基因,并将其克隆至pGM-T载体,经PCR、酶切和测序鉴定后,发现新疆株Eg95-XJ-1基因片段为1 304 bp。与Gene Bank中已知的Eg95基因有86%~99%的同源性,其差异大都集中在内含子区;与青海株Eg95基因家族成员同源性最高达97%~99%。这些结果表明,这一新疆地区的Eg95-XJ-1基因为Eg95基因家族成员之一。     

Construction of Full-length cDNA of Recombinant Rabies Virus SRV9 Strain Expressing EgM123 Gene of Echinococcus granulosus

The aim of the experiment was to construct the recombinant rabies virus SRV₉ vaccine strain with EgM123 gene by reverse genetics technology and provide the technical means for effective prevention and control of rabies and hydatidosis in China's agricultural and pastoral areas.In this study,the structural protein N,P and L genes of rabies virus SRV₉ were synthesized using gene synthesis technology,which was based on the complete genome sequence of rabies virus SRV₉ and the fusion fragment of the N-P-M fusion fragment and the rabies G gene,through the carrier of enzyme insertion connection methods,the recombinant rabies virus L gene,N-P-M gene fusion fragment and G+EgM123+eGFP gene fusion fragment were successively recombined on the expression vector pcDNA3.1(-) to construct the full-length cDNA of recombinant rabies virus SRV₉ with EgM123 gene.The synthesized genes were constructed on pcDNA3.1(-) expression vector,and the results of transformation,plasmid digestion and gene sequencing showed that the length of N,P,L,N+P+M and G+EgM123+eGFP gene fragments were 1 365,1 107,6 471,3 160 and 3 256 bp,respectively.The full-length cDNA fragment of EgM123 gene recombinant rabies virus full-length cDNA was 12 465 bp,and the sequencing results of each gene fragment were 100%.In this experiment,the full-length cDNA fragment of recombinant EgM123 rabies and eukaryotic expression vectors of the N,P and L genes of rabies virus were successfully constructed,which could save EgM123 gene recombinant rabies by reverse genetics,it also provided the reference for the development of rabies and hydatid disease combined gene recombinant oral live vaccine.     

Movement patterns of armado, Pterodoras granulosus, in the Paraná River Basin

Abstract –  We studied the migratory behaviour of armado, Pterodoras granulosus , in the Paraná River Basin of Brazil, Paraguay and Argentina, during 1997–2005. This species invaded the Upper Paraná River after upstream dispersal was facilitated when Itaipu Reservoir inundated a natural barrier. Fish were tagged ( N  = 8051) in the mainstems of the Yacyreta and Itaipu reservoirs, bays of major tributaries, the Paraná River floodplain above Itaipu Reservoir, and below dams. In all, 420 fish were recaptured of which 61% moved away from the release area. Fish moved a maximum of 215 km (mean 42), and at a maximum rate of 9.4 km·day⁻¹ (mean 0.6). Of the 256 armados that moved away from the release site, 145 moved upstream towards unimpounded stretches of the Paraná River and 111 moved downstream into the reservoir and bays of its tributaries (maximum 150 km). Based on the observed migratory movements, we suspect that most of the reproductive output originates in tributaries to the reservoirs. The ability of this species to expand its range presents a conundrum by pitting fishery management interests against conservation needs. Maintenance of the important armado fisheries depends on the ability of the species to migrate freely to use spawning and nursery areas in reservoir tributaries and floodplains. However, its ability to migrate long distances can allow this non-native species the opportunity to invade most of the Upper Paraná River.     

Cystic echinococcosis in the Falkland Islands

From 1991 to 1993, an investigation into the epidemiology of cystic echinococcosis (CE) was carried out in the Falkland Islands to evaluate the progress of the hydatid eradication campaign. The prevalence of CE in sheep was assessed using abattoir and farm slaughter data, and the exposure of dogs to the parasite was estimated using immunological techniques. A total of 59 466 sheep was examined at slaughter for E. granulosus and T. hydatigena cysts and the entire dog population of the Falkland Islands (n = 908) was examined by ELISA for the presence of specific serum antibodies to E. granulosus (IgG, IgA and IgE). In addition, a subsample of dogs (n = 464) was tested for the presence of E. granulosus antigens in faeces (copro-antigens). The prevalence of CE in sheep increased significantly during the period of the study from 0.11% in 1991 to 0.47% in 1993. Nineteen (2.1%) of 908 dog sera tested were seropositive, and eight dogs (1.7%) of 464 tested were positive in the copro-antigen assay. The combined use of abattoir surveillance, specific antibody and copro-antigen assay suggested that there were several locations in the Falkland Islands where the life cycle of E. granulosus may still perpetuate. Specific deficiencies in the eradication effort in those locations could be identified through follow-up questionnaires.     

青海牦牛和藏羊泡球蚴的感染及病理形态学观察

对青海高原地区牦牛和藏羊体内多房棘球蚴的感染进行调查,感染率分别为4.7％和5.4％;用病理组织学技术对牦牛和藏羊体内寄生的泡球蚴形态结构进行观察,证实大部分泡球蚴结构受损或退化,虽然部分结构高度分化,但不产生或极少产生原头蚴,并对其流行病学意义进行了讨论。     

木薯主要地下害虫蛴螬、蔗根锯天牛发生规律与防控技术研究

为了有效防控木薯地下害虫,笔者就木薯主要地下害虫蛴螬、蔗根锯天牛的发生规律与防控技术进行了研究。结果表明,蛴螬发生一代至少1年,成虫在6～8月雨后羽化出土,高温干旱下,坡地、沙质地及木薯连作地、甘蔗轮作地及花生间套作地受害较重;蔗根锯天牛2～3年发生一代,成虫在4～6月雨后羽化出土,高温干旱下,坡地、沙质地及木薯连作地、甘蔗轮作地受害较重;在种植前选用环保型杀虫剂48％乐斯本乳油 1 000倍液和1.8％阿维菌素乳油2 000倍液混合液浸泡种茎5～10 min后种植效果最好,可显著提高出苗率、降低虫害率和提高产量;土坑对金龟子和蔗根锯天牛也具有良好的诱杀效果;“扫虫光”不仅对蔗根锯天牛、蛴螬等木薯主要地下害虫具有良好的防治效果,而且可以促进木薯生长,显著提高鲜薯产量;以种茎药剂处理、土壤药剂处理、土坑诱杀和合理轮作为核心的木薯地下害虫环境友好综合防控效果十分显著,综合防控区平均虫害率由对照的45.6％显著下降到4.2％,产量由对照的16 920 kg/hm2显著提高到49 960 kg/hm2。上述结果为可持续防控木薯害虫的发生与危害提供了理论与技术支撑。     

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。