Li Jian, Shi Yun-Liang, Shi Wei, Fang Fang, Zhou Qing-An, Li Wen-Wen, He Guo-Sheng, Huang Wei-Yi
College of Animal Science and Technology, Guangxi University, Nanning 530005, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2012 Apr 30;30(2):81-5.
To observe the ultrastructure of nymphal Armillifer sp. isolated from Macaca fascicularis by using scanning electron microscope (SEM), and analyze the phylogenetic relationships based on 18S rRNA gene sequences.
The parasite samples stored in 70% alcohol were fixed by glutaraldehyde and osmium peroxide. Ultrastructural characters of those samples were observed under SEM. Amplification and sequencing of the 18S rRNA gene were performed following the extraction of total genome DNA. Sequence analysis was performed based on multiple alignment using ClustalX1.83, while phylogenetic analysis was made by Neighbor-Joining method using MEGA4.0.
The nymphs were in cylindrical shape, the body slightly claviform tapering to posterior end. Abdominal annuli were gradually widened from anterior to posterior parts, the 12th-13th abdominal annuli of which were similar in width. The annuli ranged closer in the front half body, whereas in the latter part there were certain gaps between them. The circular-shaped mouth located in the middle of head ventrally. Folds were seen in inner margin of the mouth with a pair of curved hooks on both sides above it which practically disposed in a straight line. Two pairs of large sensory papillae were observed symmetrically over the last thoracic annulus of cephalothoraxs lying below the outer hook, and the first abdominal annulus was near the median ventral line. The number of abdominal annuli was 29, not including 2 incomplete terminal annuli. Rounded sensory papillae were fully distributed on the body surface, except the dorsal side of head and the ventral part of the terminal annulus. Agglomerate-like anus opening was observed at the end of ventral abdominal annuli and distinctly sub-terminal. These morphological features demonstrated that the nymphs were highly similar with that of Armillifer moniliformis Diesing, 1835. A fragment of 18SrRNA gene (1 836 bp) sequences was obtained by PCR combined with sequencing, and was registered to the GeneBank database with an accession number HM048870. The phylogenetic tree indicated that A. moniliformis, A.agkistrodon and A.armillatus were at the same clade with a bootstrap value at 95%, and A. moniliformis and A. agkistrodon were solo at a clade with a bootstrap value of 75%.
The nymphs isolated from Macaca fascicularis are identified as A. moniliformis temporarily.
利用扫描电子显微镜观察从食蟹猴分离出的有钩绦虫蚴的超微结构，并基于18S rRNA基因序列分析其系统发育关系。
将保存在70%乙醇中的寄生虫样本用戊二醛和过氧化锇固定。在扫描电子显微镜下观察这些样本的超微结构特征。提取总基因组DNA后进行18S rRNA基因的扩增和测序。使用ClustalX1.83进行多序列比对后的序列分析，同时使用MEGA4.0通过邻接法进行系统发育分析。
蚴虫呈圆柱形，身体略呈棒状，向后端逐渐变细。腹部环纹从前向后逐渐变宽，其中第12 - 13个腹部环纹宽度相近。环纹在前半身排列较紧密，而后半身之间有一定间隙。圆形口位于头部腹面中央。口的内缘可见褶皱，其上方两侧有一对弯曲的钩，几乎呈直线排列。在头胸部最后一个胸环的外钩下方对称观察到两对大的感觉乳突，第一腹环靠近腹中线。腹部环纹数量为29个，不包括2个不完全的末端环纹。圆形感觉乳突遍布体表，除头部背面和末端环纹腹面部分。在腹侧腹部环纹末端观察到团聚状肛门开口，明显位于亚末端。这些形态特征表明蚴虫与1835年迪辛描述的念珠棘虫高度相似。通过PCR结合测序获得了一段18SrRNA基因（1836 bp）序列，并以登录号HM048870注册到基因库数据库中。系统发育树表明，念珠棘虫、竹叶青棘虫和有棘棘虫处于同一分支，自展值为95%，念珠棘虫和竹叶青棘虫单独处于一个分支，自展值为75%。
从食蟹猴分离出的蚴虫暂被鉴定为念珠棘虫。
