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Toxoplasma gondii rhoptry protein 18(ROP18)is a promising vaccine candidate against toxoplasmosis in mice

Toxoplasma gondii rhoptry protein 18(ROP18)is a promising va...

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中国畜牧兽医学会家畜寄生虫学分会第六次代表大会暨第十一次学术研讨会

作者： Yuan Ziguo~1,Zhang Xiuxiang~1,He Xianhui~1,Zhou Donghui~1, Lin Ruiqing~1,Zhu Xingquan~2 1 College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China 2 State Key Laboratory of Veterinary Etiological Biology,Key Laboratory of Veterinary Parasitology of Gansu Province,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences, Lanzhou 730046,China

Toxoplasma gondii is an obligate intracellular protozoan parasite infecting a large variety of domestic and wild animals and human *** are involved in *** invasion and host cell interaction,and have been implicated as important virulence *** this study,we constructed a DNA vaccine expressing rhoptry protein 18(ROP18) of ***,and evaluated the immune response it induced in Kunming *** gene sequence encoding ROP18 was inserted into the eukaryotic expression vector pVAX *** immunized Kunming mice *** immunization,we evaluated the immune response using lymphoproliferative assay,cytokine and antibody measurements,and the survival times of mice challenged *** results showed that the group immunized with pVAX-ROP18 developed a high level of specific antibody responses against *** ROP18 expressed in ***,a strong lymphoproliferative response,and significant levels of IFN-γ,IL-2,IL-4 and IL-10 production,compared with the other groups immunized with either empty plasmid or phosphate-buffered saline,respectively. These results demonstrate that pVAX-ROP18 induces significant humoral and cellular Th1 immune *** lethal challenge,the mice immunized with the pVAX-ROP18 showed a significantly increased survival time(27.9±15.1 days) compared with control mice which died within 7 days of *** data demonstrate,for the first time,that ROP18 triggered a strong humoral and cellular response against ***,and that ROP18 is a promising vaccine candidate against toxoplasmosis,worth further development.

关键词： ROP18)is a promising vaccine candidate against toxoplasmosis in mice Toxoplasma gondii rhoptry protein 18

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IRAP:An ef cient retrotransposon-based electrophoretic technique for studying genetic variability among geographical isolates of Schistosoma japonicum

IRAP:An ef cient retrotransposon-based electrophoretic techn...

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中国畜牧兽医学会家畜寄生虫学分会第六次代表大会暨第十一次学术研讨会

作者： Li Juan~(1,2),Zhao Guanghui~(1,3),Li Xiaoyan~4,Chen Fen~(1,2),Chen Jingbin~5,Zou Fengcai~6,Yang Jianfa~6, Lin Ruiqing~2,Weng Yabiao~(1,2),Zhu Xingquan~(1,6) 1 State Key Laboratory of Veterinary Etiological Biology,Lanzhou Veterinary Research Institute,CAAS, Lanzhou,730046 China 2 College of Veterinary Medicine,South China Agricultural University,510642 China 3 College of Veterinary Medicine,Northwest A & F University,Yangling,712100 China 4 College of Agriculture,Northwest A & F University,Yangling,712100 China 5 College of Agriculture,South China Agricultural University,510642 China 6 College of Animal Science and Technology,Yunnan Agricultural University,650201 China

In the present study,an inter-retrotransposon-amplified polymorphism(IRAP) technique,based on retrotransposons,was used to examine genetic variability among Schistosoma japonicum isolates from different provinces in China's *** the 15 primers screened,5 produced highly reproducible IRAP *** these primers,54 discernible DNA fragments were generated with 40(74.07%) being polymorphic,indicating considerable genetic variation among the examined *** *** primer LTR-11 was found to be able to differentiate male and female parasites,producing one constant specific band for female *** *** percentages of polymorphic bands(PPB) among all parasites,among isolates from mountainous provinces and among those from the lake/marshland areas were 74.07,48.15,and 66.67%,*** analysis revealed that the IRAP profiles could group *** isolates in China's mainland into two clades(mountainous and lake/marshland types),and samples from the same geographical origins clustered *** results demonstrated that the IRAP technique is suitable for studying genetic diversity and population structures,and also provides an effective technique for studying sex differentiation of ***.

关键词： IRAP:An ef cient retrotransposon-based electrophoretic technique for studying genetic variability among geographical isolates of

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Construction and analysis of expressed sequence tags from a cDNA library of male adult of Toxocara canis

Construction and analysis of expressed sequence tags from a ...

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中国畜牧兽医学会家畜寄生虫学分会第六次代表大会暨第十一次学术研讨会

作者： Zhou Rongqiong~(1,2),Huang Hancheng~1,Hu Shijun~1,Zhou Zuoyong~1,Xia Qingyong~2 1 Department of Veterinary Medicine,Rongchang campus,Southwest University,Chongqing 402460,P. R.China 2 Institute of Sericulture and Systems Biology,Southwest University,Chongqing 400715,P.R.China

The nematode Toxocara canis is a cosmopolitan parasite of canids,and has a wide range of paratenic hosts,including *** order to study the gene expression including development,digestion and reproduction organs of ***,a cDNA library had been constructed and expressed sequence tags (ESTs) had been *** are useful resources in annotating genes and providing functional analysis of *** this study,a cDNA library from male adult nematode of *** was created and 189 ESTs from 5'-ends of the cDNA clones representing 101 unigenes were *** titer of the primary cDNA library was 2.71×10～6 pfu/mL with a recombination rate of 99.47%.Most of the cDNA inserts ranged from 500 to 2000 bp with an average length of 1000 *** analysis of these ESTs allowed identification of 101 unique sequences containing 271 contigs and 74 *** searches revealed that 45 unigenes(44.55%of the total) or 88 ESTs(46.56%of the total) were novel genes that had no significant matches to any protein sequences in the public *** rest 56 unigenes(55.44%of the total) or 101 ESTs(53.44%of the total) were closely matched to the known genes or sequences deposited in the public *** the EST sequences were deposited under dbEST in GenBank (GenBank:HO348195-HO348295).These genes were classified into six groups based on their know or putative biological *** also confirmed the sex-specific gene expression patterns of two identied genes using RT-PCR *** work will provide a valuable resource for understanding this nematode growth,reproduction and metabolic functions.

关键词： Expressed sequence tag(EST) cDNA library Toxocara canis Male

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Immunization with a recombinant 58-kilodalton antigen from Baylisascaris schroederi induces protective immunity against larval migration of *** in mice

Immunization with a recombinant 58-kilodalton antigen from B...

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中国畜牧兽医学会家畜寄生虫学分会第六次代表大会暨第十一次学术研讨会

作者： Xie Yue,Gu Xiaobin,Fu Yan,Zhang Runhui,Zheng Wanpeng,Peng Xuerong,Wang Shuxian, Yang Guangyou~* Department of Parasitology,College of Veterinary Medicine,Sichuan Agricultural University,Ya'an 625014,China

Baylisascaris schroederi(Nematoda:Ascaridida) is the leading cause of death from primary and secondary infection in wild and captive giant pandas due to a high infection rate and the pathogenic consequences of visceral larval migrans(VLM).To date,little was known about how protective immunity to *** infection could be *** this study,we have identified a cDNA encoding B. schroederi 58-kDa(Bs58) antigen(L2,L3 and adult stages) and evaluated the immune effect of the Escherichia coli-expressed recombinant protein(rBs58) in *** analysis showed that Bs58 has no similarity at the amino acid level to mammalian proteins but has some similarity to those of the filarial parasites and Caenorhabditis elegans gene ***,an intense expression of endogenous As24 in the body wall,gut epithelium,ovary and uterus of adult female worms was determined by *** addition,Bs58 homologues were found to be expressed in human,swine and dog *** of mice in three consecutive doses with rBs58 in Freund's Complete Adjuvant (FCA) results in protection against challenge infections as manifested by a 69%reduction(P<0.001) in recovery compared with that in the non-treated group at day 7 *** obtained from immune protected mice showed a significantly increased in the level of the total immunoglobulin G(IgG) (p<0.0001) but had no IgE and IgM *** of the IgG subclasses showed a higher level of IgG1(p0.0001) and a lower level of IgG2a(p<0.005) antibody response in the sera of the immunized mice,suggesting that protection was associated with a typeⅡimmune *** immunization trial results in a *** model suggest the possibility of using rBs58 to develop a useful vaccine candidate antigen for parasitic diseases caused by ascarid nematodes in giant pandas and other animals.

关键词： Immunization with a recombinant 58-kilodalton antigen from Baylisascaris schroederi induces protective immunity against larval migration of B.schroederi in mice

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Molecular characterization and phylogenetic analysis of ascarid nematodes from Ailuropoda melanoleuca and other twenty captive wild mammals based on mitochondrial and nuclear sequences

Molecular characterization and phylogenetic analysis of asca...

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中国畜牧兽医学会家畜寄生虫学分会第六次代表大会暨第十一次学术研讨会

作者： Li Yan,Xie Yue,Gu Xiaobin,Fu Yan,Zhang Runhui, Zheng Wanpeng,Peng Xuerong,Wang Shuxian,Yang Guangyou~* Department of Parasitology,College of Veterinary Medicine,Sichuan Agricultural University,Ya'an 625014,China

Although ascarid nematodes are the most important parasites of wild animals with public health concern,most ascarids from wild animals has not been studied at molecular *** classification and phylogenetic relationships among and within roundworms from twenty-one captive wild animals in zoo were studied by sequencing and analysis of some parts of the ribosome 18S and 28S genes and mtDNA 12S *** and identity analysis of the three genes showed that roundworms from these animals were distributed into five genus:Toxocara cati taxa,Toxocara canis,Toxascaris,Baylisascaris and Ascaris. Homology analyses indicated that 18S genes sequence were conserved among roundworms from all these 21 animals.28S genes showed variability among *** levels of 12S genes sequences ranged from 11.7%to 12.8%between Toxocara cati and Toxocara canis indicating that mt 12S gene appeared variability among intraspecies and *** relationship were inferred by three methods,NJ/MP/ML based on the datas of rDNA,12S gene and combined three *** tree suggested that the ascarids of 21 species wild animals were classified into two families,four genus and seven *** alternative topologies showed that each clade was supported by high bootstrap value. Meanwhile,the combined trees suggested that the genetic relationships of Baylisascaris ailuri are closer to Baylisascaris transfuga than those of Baylisascaris schroederi and received higher support(94%).This study has provided useful molecular markers for the epidemiological investigation of roundworms from wild animals.

关键词： Molecular characterization and phylogenetic analysis of ascarid nematodes from Ailuropoda melanoleuca and other twenty captive wild mammals based on mitochondrial and nuclear sequences

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扫描电镜超微结构观察鉴定蛇舌状虫属(Armillifer spp.)若虫与基于其18SrRNA基因的种系发育关系分析

扫描电镜超微结构观察鉴定蛇舌状虫属(Armillifer spp.)若虫与基...

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中国畜牧兽医学会家畜寄生虫学分会第六次代表大会暨第十一次学术研讨会

作者：李健石云良施维方芳周庆安李雯雯何国声黄维义广西大学动物科学技术学院中国农业科学院上海兽医研究所农业部动物寄生虫学重点实验室

目的运用扫描电镜超微结构观察以鉴定食蟹猴体内分离的蛇舌状虫若虫,并基于18SrRNA序列分析其分子种系发育关系。材料方法将保存于70%酒精中的若虫应用常规的戊二醛与锇酸双固定法,置于扫描电镜下观察。消化样品,提取全基因组DNA,对18Sr... 详细信息

目的运用扫描电镜超微结构观察以鉴定食蟹猴体内分离的蛇舌状虫若虫,并基于18SrRNA序列分析其分子种系发育关系。材料方法将保存于70%酒精中的若虫应用常规的戊二醛与锇酸双固定法,置于扫描电镜下观察。消化样品,提取全基因组DNA,对18SrRNA基因进行扩增并测序;运用序列比对软件ClustalX 1.83将测序结果与相关序列进行多重比对分析,用MEGA 4.0采用邻位相连法构建种系发育树进行分析。结果若虫呈圆柱状,前段略粗,末端变细,呈纺锤形。前段腹环上下距离逐渐增宽,至12～13腹环时趋于等宽,腹环与腹环之间,在前半段连接紧密,在后半段有一定间隔。头部腹面正中为口呈圆形,口稍上方两侧各见一对钩,钩几乎在同一平行线上。两外侧钩正下方,头胸部最后一节胸环上各有一个对称的大型感觉乳突,紧接的第一节腹环靠中线处有一对对称的大型感觉乳突,腹环数目由此算起共29个腹环,末端有2个腹面未连接的不完整腹环不计算在内。若虫全身布满近圆形的感觉乳突,但在头部背面与末节腹面未见有这样的乳突出现。末端腹面有一块状隆起上可见肛门开口。大部分形态特征与串珠蛇舌状虫(Armillifer moniliformis)相似。扩增得到长度为1836bps的18SrRNA基因序列,经测序后采用邻位对接法构建的种系发育树,表明***与***、***构成自展值为95%的分支,前二者又构成一个独立的自展值为75%的分支。此次分离食蟹猴的蛇舌状虫若虫暂鉴定为串珠蛇舌状虫若虫。讨论根据此次分离到蛇舌状虫若虫的地域分布暂将其定为***。扫描电镜观察此次分离的若虫的体表特征,将其与之前张永年等报道的***若虫相比有明显不同,较Banaja报道的***若虫存在更为显著差异。值得一提的是该若虫腹环后缘形成的"锯齿样"饰物与***若虫腹环后缘的单行细刺是不一样的。以上的观察将为我国对***若虫虫种的鉴定及药物治疗等方面的研究提供基础资料。分子种系进化关系方面,本文的结果与Tappe等基于***的18SrRNA基因序列构建种系发育树关于舌形虫与其宿主间协同进化的结论吻合,而且同处于亚洲分布的***、***两个种独立于非洲分布的***能形成一个自展值为75%的拓扑分支,暗示着依据地域分布鉴定虫种的合理性。此次种系发育分析结果已足以在分子上区别于该虫与之前造成错误鉴定的***、***,为今后***若虫所致的人舌形虫病提供了分子诊断的可能。

关键词：种系发育关系 Armillifer spp 电镜超微结构观察

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柔嫩艾美耳球虫Rhomboid蛋白与微线蛋白相互作用的研究

柔嫩艾美耳球虫Rhomboid蛋白与微线蛋白相互作用的研究

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中国畜牧兽医学会家畜寄生虫学分会第六次代表大会暨第十一次学术研讨会

作者：郑君张西臣宫鹏涛杨举李赫张国才李建华吉林大学畜牧兽医学院

目的:筛选与柔嫩艾美耳球虫Rhomboid蛋白存在相互作用的微线蛋白,并进一步验证两种蛋白的相互作用,然后观察互作蛋白对雏鸡的免疫保护作用以及Rhomboid蛋白酶抑制剂体外对子孢子侵入细胞的影响。方法:构建CytoTrap酵母双杂交系统诱饵质... 详细信息

目的:筛选与柔嫩艾美耳球虫Rhomboid蛋白存在相互作用的微线蛋白,并进一步验证两种蛋白的相互作用,然后观察互作蛋白对雏鸡的免疫保护作用以及Rhomboid蛋白酶抑制剂体外对子孢子侵入细胞的影响。方法:构建CytoTrap酵母双杂交系统诱饵质粒pSos-Rho,转化酵母菌cdc25H,验证是否存在自激活作用;然后构建Ga14酵母双杂交系统诱饵质粒pGBKT7-Rho与捕获质粒pGADT7-MIC1和pGADT7-MIC4,分别共转化酵母菌AH109,筛选相互作用的蛋白;接着将互作的两种蛋白通过大肠杆菌和HeLa.细胞表达,通过免疫共沉淀进一步验证两种蛋白的相互作用;后将含有互作蛋白的酵母细胞AH109破碎后免疫雏鸡,检测体液免疫水平和细胞免疫水平,通过攻虫试验观察互作蛋白对雏鸡的免疫保护作用;最后通过体外培养***,验证Rhomboid蛋白酶抑制剂DCI的体外抑制作用。结果:Rhomboid构建的重组诱饵蛋白在CytoTrap酵母双杂交系统中存在自激活作用,而在Gal4酵母双杂交系统中,对酵母细胞既没有毒性,也不存在自激活作用,通过Gal4酵母双杂交系统的筛选,Rhomboid与MIC4蛋白存在相互作用,免疫共沉淀试验进一步证明,两种蛋白存在相互作用,并且Rhomboid蛋白可以切割其底物MIC4蛋白;将含有互作蛋白的酵母细胞AH109破碎后免疫雏鸡,与对照组相比,体液免疫水平和细胞免疫水平均差异显著(P<0.01),攻虫后抗球虫指数ACI为168.24;体外抑制试验显示,与对照组相比,第一代裂殖体数量在DCI浓度20μM和30μM时差异极显著(P<0.01),第二代裂殖体数量在DCI浓度20μM时差异显著(P<0.05),子代卵囊数在DCI浓度20μM时差异显著(P<0.05)。结论:本研究成功证明柔嫩艾美耳球虫Rhomboid蛋白与MIC4蛋白存在相互作用,且Rhomboid蛋白可以切割其底物MIC4蛋白;将含有这两种互作蛋白的酵母细胞破碎后免疫雏鸡,显示了较好的免疫保护性;20μM DCI即可有效抑制子孢子体外对鸡肾细胞的侵入,同时能够控制球虫子代卵囊的产出量,对球虫病有潜在的治疗作用。本研究为更深入的探讨柔嫩艾美耳球虫侵入宿主细胞的分子机制奠定了基础,也为球虫病的疫苗预防和药物治疗提供理论依据。

关键词：柔嫩艾美耳球虫酵母细胞酵母双杂交系统微线蛋白球虫病 DCI 互作蛋白 Rhomboid

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Characterization of Baylisascaris schroederi from Qinling subspecies of giant panda in China by the first internal transcribed spacer(ITS-1)of nuclear ribosomal DNA

Characterization of Baylisascaris schroederi from Qinling su...

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中国畜牧兽医学会家畜寄生虫学分会第六次代表大会暨第十一次学术研讨会

作者： Lin Qing,Li Hongmei,Gao Man,Wang Xingyie,Ren Wanxin,Cong Meimei,Tan Xuechao,~* Yu Sanke,~* Zhao Guanghui (College of Veterinary Medicine,Northwest A & F University,Yangling,Shaanxi Province 712100,PR China)

The VLM due to Baylisascaris schroederi infection was identified as the most significant threat to wild panda survival,which was responsible for about 50%of the *** deaths between 2001 and *** traditional approaches for the identification and differentiation based on morphological features need experts and are sometimes insufficient to distinguish closely related species,and the biological studies are usually labor-consuming,and can be affected by the status of host,treated methods, and other *** the present study,a total of 20 nematode isolates（including 10 male and 10 female worms） representing Baylisascaris schroederi from 5 Qinling subspecies of giant pandas（Ailuropoda melanoleuca） in Shaanxi province of China,were characterized and grouped genetically by the first internal transcribed spacer（ITS-1） of nuclear ribosomal DNA（rDNA）.The rDNA fragment spanning 3' end of 18S rDNA,complete ITS-1 rDNA,and 5' end of 5.8S rDNA was amplified and *** sequence variability in ITS-1 rDNA was examined within *** and among parasites in order Ascaridata available in GenBank,and their phylogenetic relationships were also *** sequences of ITS-1 rDNA for all the *** isolates were 427 bp in length,with no genetic variation detected among these *** analyses based on the ITS-1 rDNA sequences revealed that all the male and female *** isolates sequenced in the present study were posited into clade of genus Baylisascaris,sistered to zoonotic nematodes in genus Ascaris,and the ITS-1 rDNA sequence could distinguish different species in order *** results showed that the ITS-1 rDNA provides a suitable molecular marker for the inter-species phylogenetic analysis and differential identification of nematodes in order Ascaridata.

关键词： Characterization of Baylisascaris schroederi from Qinling subspecies of giant panda in China by the first internal transcribed spacer ITS-1)of nuclear ribosomal DNA

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Analysis of T cell receptor(TCR)usage in Eimeria maxima immunized SPF white leghorn chickens

Analysis of T cell receptor(TCR)usage in Eimeria maxima immu...

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中国畜牧兽医学会家畜寄生虫学分会第六次代表大会暨第十一次学术研讨会

作者： Ren Chao,Liu Xianyong,Suo Xun~* National Animal Protozoa Laboratory & College of Veterinary Medicine,China Agricultural University, Beijing 100193,China

Among seven Eimeria species that infect chickens,Eimeria maxima is the most immunogenic species and can elicit full immune protection in vaccinated *** findings suggest that immunodominant protective antigens exist in several protozoan parasites,such as GRA6 antigen derived from Toxoplasma gondii,a protozoa belonging to *** it is logically to assume that one or a few immunodominant antigens also exist in ***,which can induce the expansion of ***-specific CD8+T cell population(s) in the *** other words,one or more CD8+T cell populations expressing same T cell receptors(TCRs) could be detected in a much higher frequency in the whole CD8+T cell population. Here we endeavored to reveal whether biased TCR usage exists in TCRβrepertoire in *** infected *** white leghorn chickens were immunized orally with 3 doses of *** oocysts or just PBS as *** CD8+T cells from blood and small intestine were subjected subsequently to total RNA extraction,cDNA synthesis and PCR amplification of TCR Vβchain sequences containing complementarity-determining region 3(CDR3).We obtained 110 sequences from 3 unimmunized chickens, 90 sequences from one chicken 2 days post the third immunization and 150 sequences from 3 chickens 5 days post *** analysis of CDR3 regions revealed that there was no biased TCR usage in CD8+T cells derived from the peripheral blood mononuclear cells(PBMCs) of the unimmunized or immunized ***,the TCRβrepertoire showed biased TCR usage in the intra-epithelial CD8+lymphocytes (IELs) from the challenged birds,and these biased TCRs were also existed in other challenged *** data are needed to confirm the persistence and public usage of the predominant TCRβclonotype distribution in more challenged birds.

关键词： SPF TCR)usage in Eimeria maxima immunized SPF white leghorn chickens Analysis of T cell receptor

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Detection and identification for nematode Gnathostoma in imported Monopterus albus

Detection and identification for nematode Gnathostoma in imp...

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中国畜牧兽医学会家畜寄生虫学分会第六次代表大会暨第十一次学术研讨会

作者： Li Shuqing~1,Li Wenwen~2,Chen Zhifei~1,Li Jian~2,Chen Shaohong~3,Zhang Yongnian~3 Huang Weiyi~2 Wang Qiaoquan~1 1.Shanghai Entry-Exit Inspection and Quarantine Bureau of the People's Republic of China,Shanghai 200135 2.College of Animal Science and Technology,Guangxi University,Nanning 530005 3.Chinese Center for Disease Control and Prevention,National Institute of Parasitic Diseases, Shanghai 200025

Objective To inspect the third stage larvae of Gnathostoma in imported Monopterus albus,and identify its species..Method 10 batches of imported Monopterus albus to Shanghai from abroad were detected for nematode Gnathostoma from January 2010 to March *** 52 Monopterus albus,3～10/batch,From Philippines(25),Indonesia(24) and Bangladesh(3),dissect each Monopterus albus,then minced,protease digestion,the suspension was filtered with 10 mesh copper screen,at the last take the precipitation of the *** the complete parasites under the stereoscope,morphological identification,and calculate the infection rate and average *** extracted genomic DNA of Gnathostoma,amplified internal transcribed spacer region 2 and cytochrome c oxidase subunit 1 by PCR,then Gel electrophoresis and *** sequences were aligned with corresponding sequences in *** The third stage larvas of nematode Gnathostoma were detected in Monopterus albus from Indonesia and Philippines,infection rates were 36.0%(9/25) and 50.0% (12/24),average infectiosity were 7.77(70/9) and 2.83(34/12).There wasn't Gnathostoma in imported Monopterus albus from *** the microscope,the larvas have one cephalic bulb,4 rings hooklets on the cephalic bulb,the body surface with cross striations and small spines,the front of the body spines were obviously and density,the rear spines getting small and *** had 1 cervical papillae,4 cervical *** characteristics was similar to Gnathostoma *** results showed,ITS-2 and cox1 PCR products were 647 bp and 441 bp,with the expected *** alignment analysis showed that,two PCR products had 99%～100%consistency with Gnathostoma spinigerum ITS-2(GenBank accession number AB181155 and Z97175) and coxl (GenBank accession number AY501388,AB180099 and AB551552).Conclusion All of the Gnathostoma detected in imported Monopterus albus from Philippines and Indonesia were Gnathostoma spinigerum.

关键词： GenBank Detection and identification for nematode Gnathostoma in imported Monopterus albus

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