武汉单极虫生活史中放射孢子虫的发现及鉴定

The life cycle of Thelohanellus wuhanensis from Carassius auratus involving an Aurantiactinomyxon-type actinospore

  • 摘要: 鲫养殖中粘孢子虫病非常严重,为掌握其病原的感染传播途径,实验通过粘孢子虫生活调查研究,在底栖寡毛类苏氏尾鳃蚓体内发现了一种放射孢子虫。该放射孢子虫的孢子无孢柄;孢体顶面观和侧面观都呈近卵圆形,长(19.8±1.3)μm,宽(18.2±1.1)μm;3个极囊梨形,呈点状聚集分布在孢体顶端,极囊长(4.53±0.4)μm,宽(3.4±0.4)μm;3个尾柄几乎等长,刺状,从孢体基部向下伸展,尾柄间夹角>100°,尾柄长(195.0±15.7)μm,宽(11.5±0.8)μm。根据形态特征将其划归为橘瓣放射孢子虫集合类群。18S rDNA序列分析表明该放射孢子虫与鲫体表寄生武汉单极虫为同一物种,序列相似率为99.8%~100%。序列系统发育分析进一步发现单极虫类群中多数种类的放射孢子虫阶段主要寄生在苏氏尾鳃蚓体内。本研究首次发现和报道了鲫寄生武汉单极虫生活史中寡毛类宿主及其放射孢子虫的形态特征,为鲫粘孢子虫病生态防控提供重要的基础资料。

     

    Abstract: Thelohanellus wuhanensis is an important parasite in crucian carp culture, forming numerous plasmodia on the skin, and causing massive mortality of juvenile fish in soil ponds. During the survey of actinosporeans in oligochaetes of fish pond in Liyang (31°18'N, 119°28'E), an Aurantiactinomyxon-type actinosporean was found from the oligochaete Branchiura sowerbyi. The actinospores are not connected with each other, and possess a style-less spore body with 3 almost equal-sized, elongate caudal processes. In apical view, the spore body is spherical and three polar capsules are positioned on the top of the spore body and close to each other. There is an above 100° angle between each process. Spore body average length (19.8±1.3)μm, width (18.2±1.1)μm polar capsules (4.53±0.4)μm long, (3.4±0.4)μm wide; caudal processes (195.0±15.7)μm long, (11.5±0.8)μm wide. Fresh spores floating in water were collected with 10 μL pipette tip under microscope and stored at -20℃. Genomic DNA was extracted with MagicMag Genomic DNA micro Kit (Sangon Biotech, China) following the instruction. Partial 18S rDNA sequence was amplified with primers MyxospecF and MyxospecR (Fiala 2006). Positive PCR products were sequenced with the primers in Sangon Biotech. Homologous sequences were determined by performing a BLASTN search at NCBI. DNA sequence similarities were calculated using the MEGA 6.0 software package. On the basis of 18S rDNA sequence analysis, the actinosporeans reported here were identified as T. wuhanensis, and showed 99.8%-100% sequence similarity with T. wuhanensis (HQ613410, JQ088179, JQ690370 and JQ968687) available in GenBank. Phylogenetic analysis revealed the actinsporean stages of most taxa in the clade Thelohanellus developed in alternative host Branchiura sowerbyi.

     

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