Studies on Antagonism between Exorchis ovariolobularis and Schistosoma japonicum in Oncomelania hupensis

目的　探讨在日本血吸虫中间宿主湖北钉螺体内叶巢外睾吸虫和日本血吸虫的对抗性。　方法　通过叶巢外睾吸虫和日本血吸虫对湖北钉螺的双重感染 ,计算血吸虫的感染率和尾蚴逸出量 ,常规石蜡切片观察钉螺组织学变化。　结果　钉螺在感染血吸虫 37d后再感染外睾吸虫 ,经一定时间后检查发现 ,血吸虫的感染率为 5 2 9% ,显著低于同时单独感染日本血吸虫对照组的感染率 (75 9% ) ;先感染叶巢外睾吸虫 ,经 10、 32、6 0、 10 0和 12 0d不同的时间间隔后再感染日本血吸虫的钉螺 ,血吸虫感染率分别为 6 4 %、 6 6 7%、 6 5 2 %、5 6 4 %和 5 7 1% ,而单独感染日本血吸虫对照组钉螺血吸虫感染率为 90 5 % ,经统计检验 ,各双重感染实验组血吸虫与对照组的感染率间差异具有显著或非常显著性意义 (P <0 0 5或P <0 0 1)。对各试验组及对照组钉螺逸出尾蚴试验发现 ,试验组钉螺血吸虫尾蚴逸出量均显著低于对照组的逸出量。组织学观察发现各双重感染组钉螺消化腺萎缩 ,消化腺盲囊间隙只有少量血吸虫子胞蚴 ,血吸虫子胞蚴皱缩、变形及不规则 ,且子胞蚴中只含稀疏的尾蚴胚球 ,有的子胞蚴中已无胚球 ;而单独感染血吸虫的对照组中血吸虫均发育到成熟的子胞蚴或尾蚴。　结论　钉螺体内叶巢外睾吸虫和日本血吸虫之间存Objective To examine the antagonistic interactions between Schistosoma japonicum and Exorchis ovariolobularis in the snail host, Oncomelania hupensis . Methods The infection rate and average number of cercariae of S japonicum were examined in O hupensis which had been superinfected with S japonicum and E ovariolobularis .The experimental snails were histologically observed by paraffin section. Results When snails were fed with eggs of E ovariolobularis 37 days later than S japonicum infection, the infection rate of S japonicum (52 9%) was significantly lower than that of the single S japonicum infected control (75 9%). When E ovariolobularis infected snails were exposed to S japonicum miracidia at different intervals, the infection rates of S japonicum were 64%, 66 7%, 65 2%, 56 4% and 57 1%, respectively. On the other hand, the infection rate of S japonicum in the single infected control was 90 5%. There existed significant differences. Furthermore, the average number of S japonicum cercariae shed from superinfected snails were significantly less than that of single S japonicum infected control. The histological observation showed that the larvae of S japonicum in superinfected snails were sluggish in development, irregular, shrunken, deformed, relatively small in number as compared with the control. The sporocysts contained few germ balls or immature cercariae and most of the digestive glands of superinfected snails were atrophied. Conclusion There is antagonism between E ovariolobularis and S japonicum in O hupensis , and E ovariolobularis is dominant in this situation

Comparison of four promoters for transient expression of RFP reporter gene in cultured Bombyx mori cells (Bm-e-HNU5)

以红色荧光蛋白基因(RFP)为报告基因,构建含4种不同启动子的重组表达质粒,用脂质体介导法转染家蚕Bombyxmori细胞(Bm-e-HNU5),观察家蚕细胞质肌动蛋白4基因启动子(A4)、α微管蛋白基因启动子(α-tub)、蚕丝心蛋白重链基因启动子(Fib)和家蚕核型多角体病毒早期即刻蛋白基因启动子(IE)4种启动子调控RFP报告基因在家蚕细胞内的瞬时表达情况。构建的重组表达质粒pDsRed-α-tub、pDsRed-A4、pDsRed-IE和pDsRed-Fib经双酶切和PCR鉴定正确无误。转染和转录实验结果表明,除了pDsRed-A4外,其他3种重组质粒在Bm-e-HNU5细胞中都得到高转染率,α-tub、IE和Fib可依次增强RFP报告基因在家蚕细胞内的瞬时表达活性。The red fluorescent protein reporter gene (RFP) was used to construct recombinant plasmids containing four different promoters, i. e., the cytoplasmic actin4 promoter (A4), α-tubulin promoter (α-tub)from silkworm, the Bombyx mori nuclear polyhedrosis virus immediate early protein promoter (IE) and the fibroin heavy chain gene promoter (Fib), respectively. These recombinant plasmids, i. e., pDsRed-A4,pDsRed-α-tub,pDsRed-Fib and pDsRed-IE, had been constructed successfully by restriction enzyme digestion and PCR analysis, and then were transfected into B. mori cell lines (Bm-e-HNU5) by lipid-mediated method to observe the ability of the four promoters to drive RFP reporter gene transient expression in cells. Transfection and transcription experiments indicated that except pDsRed-A4, the other three kinds of recombinant plasmids all transfected Bm-e-HNU5 obviously. The promoters of α-tub, IE and Fib enhanced the transient expression activity of RFP reporter gene in the Bm-e-HNU5, and their activity strengthened sequentially.国家自然科学基金项目(39870410

OBSERVATIONS ON THELIFE CYCLE OF EXORCHIS OVARIOLOBULARIS AND ITS DEVEL OPMENT IN THE NATURAL MOLLUSCAN HOST

本文报告叶巢外睾吸虫 ( Exorchis ovariolobularis)幼虫期人工感染图氏窄口螺 ( Stenothyratoucheana) ,在其体中发育的全过程。从实验螺获得的本吸虫成熟尾蚴 ,在实验室内经实验小鱼获得囊蚴。用囊蚴感染阴性鲶鱼 ( Parasilurusasotus) ,获得本吸虫的童虫和成虫。由此证明图氏窄口螺是本吸虫的适宜的贝类宿主。叶巢外睾吸虫是寄生在闽江下游的鲶鱼肠道 ,闽江两岸河畔有大量图氏窄口螺孳生栖息 ,但没有钉螺。已故唐仲璋教授早年在福州闽江已发现鲶鱼肠中的外睾吸虫成虫及在图氏窄口螺体中的本吸虫尾蚴 ,绘有草图 ,并推测该尾蚴可能是外睾吸虫的幼虫期。本文实验证实了他的推断。证明图氏窄口螺是福建闽江鲶鱼的叶巢外睾吸虫的天然贝类宿主。该地区本吸虫天然流行的具体情况有待进一步调查Observations on the experimental infection of Exorchis ovariolobularis in Stenothyra toucheana and the whole life cycle of this trematode species were repsrted.The mature cercariae were obtained from the experimentally infected snails, and then were transformed into metacercariae in experimental fishes.The metacercariae were fed to negative Parasilurus asotus, and juveniles and adults could be obtained from P.asotus's intestine. This experiment demonstrated that S.toucheana is an appropriate molluscan host of this trematode species. E.ovariolobularis parasitizes the intestine of Parasilurus asotus of lower reaches of Min River. On the sides of lower reaches of Min River, There are many snails of S.toucheana which is the major species in that enviornment where no Oncomelania hupensis can be found.The late professor Tang C.C.had found adults of E.ovariolobularis in the intestine of Parasilurus asotus and cercaria of this species of tremotode in S.toucheana. He supposed that this cercaria should be the larval stage of E.ovariolobularis. This experiment confirmed his conjecture and demonstrated that S.toucheana is the natural molluscan host of E.ovariolobularis. The natural epidemiology of E.ovariolobularis needs further investigations

The histology, histochemistry and scanning electron microscopy study of Exorchis ovariolobularis rediae

运用组织学、组织化学及扫描电镜方法对叶巢外睾吸虫 ( Exorchis ovariolobularis)雷蚴进行了详细的观察 .叶巢外睾吸虫雷蚴具有完善的摄食器官及强的活动能力 ,其强大的肌质咽在早期雷蚴首先分化出来 .这是在钉螺媒介中叶巢外睾吸虫直接对抗血吸虫幼虫期的物质基础 .The biological characheristics of the rediae of Exorchis ovariolobularis have been studied in the way of histology, histochemistry and scanning electron microscopy. It was found that the Exorchis ovariolobularis rediae, which is equipped with a well developed mouth, a strong muscular pharynx is actively mobile. The structure of the rediae form the basis of redial direct antagonism against Schistisoma sporocyst

Advances in the research on silkworm microsatellite DNA.

概述了分子标记技术,就其中的微卫星标记技术的原理、方法与特点进行了介绍,重点就该技术在家蚕中的研究进展进行了综述,并展望了该技术在家蚕的遗传分析、辅助选择育种等应用中的前景。In this article,the molecular marker technology is outlined and the application principle,method and traits of microsatellite marker of silkworm are reviewed.Especially,we introduce the applications of the technology in the Bombyx mori in detail and the outlook of genetic analysis,marker-assisted selection and so on are prospected

THE PATHOGENICITY OF GNATHOSTOMA HISPIDUM TO SWINE

猪颚口线虫病的临床症状是病猪经常呕吐、食欲减退、消瘦和体重减轻 ,早期血检嗜酸性细胞增高。本病的主要致病器官是胃和肝脏。胃的主要病变是胃底布满虫洞、粘膜增厚、发炎和溃疡。肝的病变是结缔组织增生、虫道出血、肝细胞索紊乱、肝细胞脂肪变性、萎缩和坏死。文中讨论了幼虫在体内的移行途径和本病的防治问题。The typical symptom of Gnathostomiasis is that the infected swine keep vomiting,anorexia,emaciated,lose weight and its eosinophil leukocyte increased.Swine stomach and liver are the main organ damaged.The main pathological changes is that the bottom of stomach is punctured by the parasite,its mucosa become thicker,inflamed and ulcer.The connected tissue of liver proliferated.The pathway along which the parasite migrate become hemorrhage,with the liver cell cord disordered.The liver cell fatty degenerated,withered and become necrosis.The way of the larve of Gnathostoma hispidum migrating in the body of swine and the control of Gnathostomiasis are discussed in this paper

PHYLOGENETIC STUDY ON EURYTREMA FROM DOMESTIC AND WILD ANIMAL

本文对普遍寄生于家畜及野生动物胰脏并引起胰吸虫病的几种阔盘属(Eurytrema)吸虫:腔阔盘吸虫(Eurytremacoelomaticum)、胰阔盘吸虫(Eurytremapancreaticun)、枝睾阔盘吸虫(Eurytremacladorchis)及福建阔盘吸虫(Eurytremafukienensis)的系统发生关系进行比较研究。分别提取寄生于不同种类宿主的这4种胰吸虫的基因组DNA ,利用随机引物进行PCR扩增,从80个随机引物中筛选出重复性好、谱带清晰并呈多态性的引物31个,共扩增出30 3条DNA带。根据重复性好、谱带清晰并呈多态性的引物扩增的结果计算遗传距离,根据遗传距离利用UPGMA方法进行聚类分析,构建出阔盘属几种吸虫的系统发生树,分子系统发生与形态学分类一致。This paper reported the phylogenetic relationship of species from Eurytrema.Genomic DNA of the six kinds of Eurytrema was extracted after identification and quantitative analysis.The RAPD-PCR was performed and screen those primers which could amplify clear and repeating bands.In 80 arbitrary primers,31 of them could amplify clear and repeating bands.A total of 303 fragments were obtained.The amplifying results was analysis by unweighted pair-group method with arithmetical averages(UPGMA).The phylogenetic tree by UPGMA are reconstructed based on genetic distance data,which is paralleled with the morphological classification.福建省自然科学基金 (B0 110 0 0 8)资助项

Genomic polymorphic DNA of five Eurytrema from domestic animal and wild animal

目的　对引起家畜及野生动物胰吸虫病的阔盘属吸虫 :腔阔盘吸虫 ,胰阔盘吸虫及分别来自牛、羊和野生动物山麂的枝睾阔盘吸虫基因组DNA多态性进行比较分析。方法　分别提取这五类胰吸虫的基因组DNA ,利用 10碱基随机引物进行RAPD -PCR扩增反应 ,选取重复性好、谱带清晰并呈多态性的引物 ,根据这些引物扩增的谱带利用UPGMA方法进行聚类分析。结果　从 6 0个随机引物中筛选出重复性好、谱带清晰并呈多态性的引物 2 2个 ,共扩增出 199条DNA带 ,其中多态性位点 114个 ,占 5 7 3% ,羊枝睾阔盘吸虫和麂枝睾阔盘吸虫的遗传距离最近 ,腔阔盘吸虫和麂枝睾阔盘吸虫的遗传距离最远。结论　寄生在野生动物山麂的枝睾阔盘吸虫与寄生在家养动物的几种胰吸虫具有很高的DNA多态性Objective To analysis the genomic polymorphic DNA of Eurytremapancreaticun,E.coelomaticum and E.cladorchis which differently from cattle,goat and Muntiacus muntjak.Methods Genomic DNA of the five kinds of eurytrema was performed and screen those primers which could amplify clear and repeating bands.The amplifying averages(UPGMA).Results In 60 arbitrary primers,22 of them could amplify clear and repeating bands.A total of 199 fragments were obtained,among which 57.3% were polymorphic.Conclusion There are high genomic DNA polymorphism among E.cladorchis which parasite on Muntiacus muntjak and those common eurytrema species which parasite on domestic animal.福建省自然科学基金 (B0 110 0 0 8)资助项