牦牛脑多头蚴病药物治疗试验

用丙硫咪唑和吡喹酮两种驱虫药物,采用"药物增量法"和"药物联合法"对24头具有明显临床症状的脑多头蚴病例牦牛进行不同剂量治疗试验.结果表明,吡喹酮增量法治疗后症状消失时间最短,为8.5 d;丙硫咪唑增量法有效率和治愈率均为100%,吡喹酮增量法的有效率80%,治愈率70%;丙硫咪唑和吡喹酮联合法的有效率67%,治愈率50%.对照组10头脑多头蚴病牦牛均在不同时间内死亡.     

湖南省山羊脑多头蚴的线粒体nad1和nad4基因的序列测定及种系发育分析

本研究旨在阐明脑多头蚴湖南分离株线粒体烟酰胺腺嘌呤二核苷酸( NADH)脱氢酶亚单位1基因(nad1)部分序列(pnad1)和烟酰胺腺嘌呤二核苷酸(NADH)脱氢酶亚单位4基因(nad4)部分序列(pnad4)的遗传变异情况,并用pnad1和pnad4序列重构脑多头蚴与其它带科绦虫的种群遗传关系.利用聚合酶链反应(PCR)扩增脑多头蚴的pnad1和pnad4,应用ClustalX 1.81程序对序列进行比对,再用Phylip3.67程序MP法和Mage4.0程序NJ法绘制种系发育树,并用Puzzle5.2程序构建最大似然树,同时利用DNAstar5.0中的Megalign程序进行同源性分析.结果显示所获得的pnad1和pnad4序列长度分别均为666和887 bp,湖南分离株与已知多头带绦虫位于同一分枝.由于脑多头蚴pnad1和pnad4序列种内相对保守,种间差异较大,故均可作为种间遗传变异研究的标记,从而为脑多头蚴的分子流行病学和其相关疾病的诊断奠定基础.     

Development of a rapid assay for the diagnosis of Myxobolus cerebralis in fish and oligochaetes using loop-mediated isothermal amplification

A loop-mediated isothermal amplification assay was developed for the rapid detection of Myxobolus cerebralis in both fish and oligochaete hosts. The assay was optimized to amplify parasitic DNA by incubation with Bst DNA polymerase and a set of six specially constructed primers at 65 degrees C for 60 min. The amplification products were detected visually using SYBR Green I dye which gave identical results to gel electrophoresis analysis. Parasite DNA was detected from infected oligochaetes, and from the anal fin, caudal fin, dorsal fin and operculum of clinically infected fish. This 'Myxo-LAMP' assay has a detection limit similar to that of a polymerase chain reaction assay (10(-6)), but is more rapid and only requires a water bath for amplification and is therefore practical for simple and rapid diagnosis of infected tissue.     

Myxobolid parasites infecting the gills of the Iberian nase, Chondrostoma polylepis (Steindachner), in Galicia (NW Spain), with a description of Myxobolus gallaicus sp. nov.

Three species of myxobolid parasites were found infecting the gills of the Iberian nase, Chondrostoma polylepis , in the River Ulla (Galicia, NW Spain), Myxobolus leuciscini , M. impressus and M. gallaicus sp. nov., which is described herein. Myxobolus leuciscini locates in the intralamellar space and the plasmodium begins to develop in the endothelium of the capillaries of the secondary lamellae. Myxobolus impressus plasmodia are located in the interlamellar multilayered epithelium. Myxobolus gallaicus plasmodia are elongated and are located exclusively in the central venous sinus of the branchial filament, suggesting that they begin their development in branchial endothelial tissue. We also present data on spore size and morphology, plasmodium and pansporoblast characteristics and pathological effects of the three species in the host branchial tissues.     

Complete developmental cycle of Myxobolus pseudodispar (Gorbunova) (Myxosporea: Myxobolidae)

Myxobolus pseudodispar (Gorbunova) is a common parasite of the muscle of roach, Rutilus rutilus L., whereas its actinosporean development occurs in two oligochaete alternate hosts. This paper reports the complete developmental cycle of this parasite in the oligochaete alternate host Tubifex tubifex and the roach. In laboratory experiments, parasite-free T. tubifex specimens were infected by myxospores of M. pseudodispar collected from roach in Lake Balaton. Parasite-free roach fingerlings were infected with floating triactinospores (TAMs) released from oligochaetes on day 69 after challenge. Young plasmodia and spores in roach were first recorded on day 80 post-exposure (p.e.). Myxospores collected from experimentally infected roach initiated a new development in T. tubifex and the resulting TAMs infected roach. No infection of roach resulted from feeding oligochaetes containing mature triactinospores.     

异育银鲫咽碘泡虫病组织病理与病理生理

咽碘泡虫(Myxobolus pharynae)病是近几年发生在江苏省盐城地区的大丰、射阳和滨海以及周边地区,引起养殖异育银鲫(Carassius auratus gibelio)大批死亡的一种黏孢子虫病,咽碘泡虫只特异地寄生在异育银鲫的咽部组织内,为了阐明该病对鱼体的损伤作用,我们对不同患病时期的异育银鲫的组织病理和疾病中期的病理生理进行研究。组织病理结果表明:疾病初期病鱼咽部略有轻度充血,咽碘泡虫以营养体阶段寄生在咽部黏膜下层的组织中,并开始形成由成纤维细胞包裹的小孢囊,其他组织器官无病理损伤现象;疾病中期由于小孢囊数量增加和囊内营养体分裂增殖并逐步发育为成熟孢子后体积增大,构成的大孢囊使咽部显著膨大,包裹小孢囊的结缔组织囊壁充血,逐步萎缩而变薄,成纤维细胞核固缩坏死,咽部黏膜层中的上皮细胞淡染、核固缩坏死,味蕾失去应有的结构,鳃组织细胞在鳃小片间严重增生,肾部分区域出现细胞坏死,肾小球肿胀,肾小管上皮细胞出现滴状玻璃样变性,肝、脾、肠和前肾无病理变化现象;疾病后期小孢囊囊壁、黏膜下层和黏膜层组织细胞进一步坏死崩解,出现孔洞,成熟孢子、坏死组织和血液一并流出孔洞,病鱼肠腔中有许多来自坏死的咽部组织细胞和成熟孢子,其他组织器官病理变化与疾病中期相似。疾病中期病鱼的病理生理分析结果表明:病鱼红细胞数量和大小、血红蛋白浓度、血栓细胞数量、血清总蛋白、白蛋白、球蛋白、血糖、谷丙转氨酶、谷草转氨酶和碱性磷酸酶都分别极显著低于健康鱼(P0.01),出现贫血;病鱼红细胞脆性、白细胞数、嗜中性粒细胞数、单核细胞数、总胆红素、肌酐、尿素和乳酸脱氢酶分别都极显著高于健康鱼(P0.01);淋巴细胞数和嗜酸性粒细胞数与健康鱼相比,无显著差异(P0.05)。由于咽碘泡虫的寄生部位鱼咽部严重膨大堵塞口咽腔并引发鳃组织增生、肾的损伤和贫血等变化,进一步发展为咽部组织坏死破裂出现孔洞流血,导致病鱼无法摄食和呼吸困难等功能障碍而死亡。     

The susceptibility of diverse species of cultured oligochaetes to the fish parasite Myxobolus pseudodispar Gorbunova (Myxozoa)

This study provides detailed information on the invertebrate hosts of Myxobolus pseudodispar (Myxozoa) and explores the susceptibility range of several species and analyses the relevance of the species composition of an oligochaete population. Our findings demonstrate that the oligochaete host range of M. pseudodispar is similarly wide as the number of vertebrate host species. Besides Tubifex tubifex and Limnodrilus hoffmeisteri, Psammoryctides barbatus and Psammoryctides moravicus were also found to be susceptible invertebrate hosts. The genetic characterization of the mitochondrial 16S rDNA of T. tubifex sensu lato revealed that lineages I, II and III are susceptible to M. pseudodispar, whereas T. tubifex lineage VI seems to be non‐susceptible. T. tubifex lineage V and L. hoffmeisteri specimens were positive in a M. pseudodispar‐specific PCR, but in most cases, the release of mature actinospores could not be detected. Hence, these non‐susceptible oligochaetes likely serve as `biological filters` as they remove myxospores from the sediment without producing actinospores. Together with the phylogenetic analysis of the susceptible and non‐susceptible oligochaete hosts on the basis of mt 16S rDNA sequences, the route of the development of M. pseudodispar in the oligochaete hosts was tracked by in situ hybridization. According to our findings, the gut epithelia seem to be a portal of entry of the sporoplasms, where the development of the parasite also takes place. The basal lamina seems to be involved in the migration of the parasite, and the worm’s cellular immune response is activated by the infection.