三棱碘泡虫 (粘体动物门：碘泡科) 重描述及其系统发育关系分析

为有效鉴定三棱碘泡虫的有效性,补充其生物学特征,实验利用现代粘孢子虫整合分类学方法,包括形态特征、寄生特性(宿主特征和寄生部位)及分子特征,对其进行重新描述,证实其分类有效性,探讨其系统发育关系。结果显示,三棱碘泡虫专性寄生于草鱼鳃丝间,未在其他器官(肝脏、肾脏、肠道等)检获孢子。包囊呈乳白色,长椭圆形,长(2.4±0.3)(2.1～2.7) mm,宽(0.8±0.1)(0.6～0.9) mm。成熟孢子壳面观呈卵圆形,孢子后端未见"V"形褶皱,孢子外包裹有一层透明的圆形黏液膜,缝面观呈纺锤形,孢子缝面可见三条平行而突出的脊线;孢质均匀,孢子后端具有一个大嗜碘泡;成熟孢子长(10.3±0.4)(9.4～11.0)μm,宽(9.5±0.5)(8.7～10.9)μm,厚(7.4±0.5)(6.4～8.0)μm;两极囊等大,梨形,极丝7～8圈;组织病理显示,病灶鳃组织未出现严重炎症反应。BLAST比对发现,三棱碘泡虫与隐杆碘泡虫小核糖体RNA序列相似性最高(89.58%),但远低于种内序列相似性。分子系统发育分析结果显示,三棱碘泡虫先与寄生于鲤科鱼类的隐杆碘泡虫形成姊妹关系,再与寄生于胭脂鱼科鱼类鳃的诺布尔碘泡虫、微小碘泡虫及毕斯塔斯碘泡虫形成独立的鳃寄生碘泡虫分枝,表明宿主亲缘关系与组织趋向性在鱼类组织寄生碘泡科粘孢子虫系统演化历程中扮演重要作用。本研究补充了三棱碘泡虫包囊形态结构、组织趋向性及分子数据,证实了其分类有效性。     

洪湖碘泡虫PCR检测方法的建立与初步应用

为建立一种灵敏、特异的快速检测异育银鲫寄生洪湖碘泡虫(Myxobolus honghuensis)的方法,本研究根据洪湖碘泡虫ITS-5.8S r DNA基因序列筛选出一对特异性引物Mh F/R,建立PCR检测方法,对反应条件进行优化,并通过特异性试验、灵敏性试验与临床检测验证其可行性。结果显示,建立的PCR检测方法能特异性扩增洪湖碘泡虫相应的基因片段,长度为479 bp,而对试验中其他9种粘孢子虫的扩增结果均为阴性;最低能检测0.1 pg的虫体基因组DNA。通过临床样品检测,PCR方法比显微镜检测的检出率提高了19.5%。结果表明,该PCR方法特异、灵敏,适用于洪湖碘泡虫的快速检测。     

鲶病原粘孢子虫米氏碘泡虫不同株系的比较研究

【目的】研究米氏碘泡虫（Myxobolus miyairii Kudo, 1919）不同寄生部位和不同地理分布的株系差异和遗传分化情况。【方法】基于形态特征、组织向性、地理分布、18S rDNA序列相似度、遗传距离、系统发育,对米氏碘泡虫各株系进行比较分析。【结果】米氏碘泡虫各株系孢子形态特征基本一致;米氏碘泡虫重庆株系1（鲶鳃腔膜寄生）,重庆株系2（鲶肠寄生）和江西株系（鲶肠寄生）间相似度为98.6%~99.9%,遗传距离为0.000~0.013;系统发育分析显示,米氏碘泡虫重庆株系2先与江西株系聚支,其形成的进化支再与重庆株系1形成姐妹群关系。【结论】序列比较和系统发育分析结果表明,米氏碘泡虫并没有形成地理种群特有的单系,而是依据寄生部位形成肠寄生支系和鳃腔膜寄生支系。宿主种类相同的条件下,较之于地理隔离,寄生部位差异对于米氏碘泡虫种群分化的影响更大。【意义】本文米氏碘泡虫各株系的比较研究及其结果,为人们进一步了解该寄生虫的进化生物学特征积累基础资料。     

引起池养异育银鲫高死亡率的粘孢子虫新种咽碘泡虫形态和分子分析

对近几年发生在江苏省盐城地区引起池养异育银鲫大批死亡的粘孢子虫进行了形态和分子等方面分析研究,发现该粘孢子虫主要特点是:孢子梨形,部分孢子后部外周包围有透明无色的膜状鞘和壳瓣底部内侧呈现1～6个"V"形缺刻,两个极囊大小不等,孢子前端内侧两个极囊间有一个细长的囊间突起,胚质中无嗜碘泡,孢囊大小10～25 mm,孢子长16.82±0.43(16.03～17.69)μm,孢子宽10.26±0.43(9.12～10.88)μm,孢子厚8.09±0.29(7.50～8.75)μm,孢子长宽比1.64±0.09(1.50～1.93);大极囊长8.66±0.25(8.25～9.16)μm、宽3.65±0.25(3.01～4.19)μm,小极囊长8.35±0.28(7.60～8.85)μm、宽3.58±0.23(3.09～4.11)μm,囊间突起长1.80±0.12(1.59～2.00)μm。特异性地寄生在异育银鲫的口咽腔上颚的咽组织内、具有寄主和寄生在咽部的专一性,PCR扩增得到1 576 bp的18S rDNA序列,GenBank登录号为JQ726700。该粘孢子虫与相近的其它粘孢子虫比较,孢子长分别与白鲟碘泡虫(Myxobolus psephurusi)和扭曲碘泡虫(M.twistus)无显著差异(P>0.05),与M.ampullicapsulatus、M.wulii和鳙碘泡虫(M.aristichthydis)分别有显著差异(P<0.05);孢子宽和厚、极囊长和宽分别与M.ampullicapsulatus、M.wulii、白鲟碘孢虫(M.psephurusi)、鳙碘泡虫(M.twistus)和扭曲碘孢虫(M.aristichthydis)有显著差异(P<0.05);分子系统树分析结果显示,与M.ampullicapsulatus在同一个分支中,亲缘关系最近。从孢子形态学、寄主的特异性、寄生部位的专一性以及18SrDNA序列等特点综合比较分析,表明该粘孢子虫为一新种,命名为咽碘泡虫Myxobolus pharynae n.sp.。     

金鱼(Carassius auratus)咽部寄生洪湖碘泡虫(Myxobolus honghuensis)的分子鉴定和系统发育分析

在福州地区开展金鱼病害监测调查中,发现一种寄生于金鱼(Carassius auratus)咽部的粘孢子虫.为了明确虫株,本文以该粘孢子虫为研究对象,使用18S rDNA和ITS-5.8S 基因对其进行分子鉴定及系统发育树的构建和分析.结果表明,虫株形态与洪湖碘泡虫(Myxobolus honghuensis)相似,18...     

长臂虾亚科9个种系统发育关系的16S rDNA序列分析

分析了日本沼虾(Macrobrachium nipponense)线粒体基因16S rDNA部分序列,并将其与GenBank中的相关4属8个种的16S rDNA序列进行了同源性比对,探讨其系统发育关系。结果显示,在分析的442个比对位点中,变异位点188个,简约信息位点125个,碱基转换与颠换值比为1.134。以鼓虾(Alpheus cylindricus)为外群,分别用MP法、NJ法及ML法构建的长臂虾亚科4属的系统发育关系分支图显示:白虾属(Exopalaemon)首先与长臂虾属(Palaemon)以及小长臂虾属(Palaemonetes)聚在一起,然后与沼虾属(Macrobrachium)聚为一支;长臂虾属和小长臂虾属出现4个种混合相聚的现象;属间结点置信值以及聚集情况稳定。     

Myxobolus erythrophthalmi sp. n. and Myxobolus shaharomae sp. n. (Myxozoa: Myxobolidae) from the internal organs of rudd, Scardinius erythrophthalmus (L.), and bleak, Alburnus alburnus (L.)

During a survey of myxosporean parasites of cyprinid fish in Hungary, infections caused by unknown Myxobolus spp. were found in the internal organs of rudd, Scardinius erythrophthalmus, and bleak, Alburnus alburnus . Small plasmodia developed in blood vessels of the kidney, liver, testes and intestinal wall. The parasites were studied on the basis of spore morphology and by histological and molecular methods. In most cases, plasmodia were surrounded by host tissue without a host reaction; however, in advanced cases, a connective tissue capsule was seen around plasmodia. Spores collected from the two fish species differed from each other and from the known Myxobolus spp. both in their morphology and 18S rDNA sequences. The two species, described as M. erythrophthalmi sp. n. from rudd and M. shaharomae sp. n. from bleak, are characterized by a specific histotropism to blood vessels, while the organ specificity involves the kidney and for the latter species, most internal organs.     

Morphological and molecular biological studies on intramuscular Myxobolus spp. of cyprinid fish

The validity of Myxobolus species infecting the skeletal muscles of six cyprinid fish species was studied by morphological and molecular biological methods. Intracellularly developing Myxobolus spores identified as M. cyprini from the common carp, M. musculi from the barbel, and M. pseudodispar from the roach, rudd, common bream and white bream were very similar in their shape and size. Nonetheless, in species identified as M. pseudodispar, the occurrence of spores with an asymmetrical shape was higher than in M. cyprini, while asymmetrical spores were only occasionally found in M. musculi. The DNA sequence analysis of the polymerase chain reaction (PCR)‐amplified 18S rRNA gene of Myxobolus spores from these fish showed a similar phylogeny to that of their host species. As morphological studies and DNA sequence analysis demonstrated slight but real differences in the spores infecting muscles of the six cyprinid species, it is suggested that M. musculi, M. pseudodispar and M. cyprini are valid species.     

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.     

Myxobolus neurophilus Guilford 1963 (Myxosporea: Myxobolidae): a common parasite infecting yellow perch Perca flavescens (Mitchell, 1814) in Saskatchewan,Canada

The goal of this study was to identify a myxosporidian parasite infecting the central nervous system of yellow perch Perca flavescens (Mitchell, 1814) observed while investigating a fish kill in Saskatchewan, Canada. Fish were collected from seven different lakes, from two distinct watersheds. Sixty-four per cent (54/86) of yellow perch contained myxozoan pseudocysts located throughout the spinal cord and brain. Myxospores measured 16.5 μm (range 16.2–16.8) long and 8.2 μm (range 7.9–8.4) wide and contained two pyriform, mildly dissymmetrical, polar capsules measuring 7.7 μm (range 7.3–8.1) long and 2.7 μm (range 2.4–3.0) wide. The polar capsules each contained a single polar filament, with 7–9 turns per polar filament coil. Sequencing of the 18S SSU rDNA gene demonstrated >99% similarity to Myxobolus neurophilus. In 60% of infected fish, there was a mild to moderate, non-suppurative myelitis or encephalitis, or both, associated with myxospores. Axonal degeneration was present in rare cases. These findings extend the geographical distribution of M. neurophilus and suggest it may be widespread in yellow perch populations in Saskatchewan.