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基于ITS1-5.8S-ITS2 rDNA片段对印度克什米尔山谷中某物种的形态学和分子鉴定及其系统发育分析

Morphological and molecular identification of in and its phylogenetic analysis based on ITS1-5.8S-ITS2 rDNA fragment in Kashmir valley, India.

作者信息

Ara Ishrat, Khan Humira Rashid, Tanveer Syed, Fatima Nida I

机构信息

Department of Zoology, University of Kashmir, Srinagar, 190006 India.

Department of Biotechnology, University of Kashmir, Srinagar, 190006 India.

出版信息

J Parasit Dis. 2024 Dec;48(4):904-916. doi: 10.1007/s12639-024-01724-9. Epub 2024 Aug 16.

Abstract

is one of the most common round worms infecting wide range of terrestrial birds including domestic fowl. and are the species of same genus which infect other hosts predominantly aquatic birds, but show morphological similarities mainly females of these species are difficult to identify. Here, we described the morphologically distinct features of along with identification through molecular analysis using ITS1-5.8S-ITS2 ribosomal DNA. Four hundred gastrointestinal tracts of domestic fowl were analysed for a period of 2 years. Light microscopy and Scanning electron microscopy (SEM), was used for determining the morphometric measurements and characteristic morphological features of . A PCR based DNA sequencing of ITS1-5.8S-ITS2 ribosomal DNA was used for molecular characterization and phylogenetic analysis was carried out through MEGA 11. Ninety gastrointestinal tracts were found to be infected with . The male worm displayed well developed lateral and caudal alae, caudal end had an anal sucker, 12 pairs of caudal papillae and cloaca from which 2 unequal spicules emerged. Female had visible vaginal bends, vulva present approximately in the middle of the body, and caudal end had a long pointed tail. Sequence analysis revealed that the PCR products of ITS1-5.8S-ITS2 rDNA were 600 bp in size and showed 96-98% similarity with the ITS1-5.8S-ITS2 sequences of on BLAST analysis. Our ITS sequence of showed a nucleotide divergence of 4.9% with of Lucknow India. The phylogenetic analysis clearly differentiated from the other representatives of the genus and revealed the relationship among different species. The study revealed the infection status of prevailing among domestic fowl, and need of strategic control measures. The results of the study suggest that ITS rDNA would serve as a potential molecular marker in identification, diagnosis and systematics of species.

摘要

是感染包括家禽在内的多种陆生鸟类的最常见蛔虫之一。和是同一属的物种,主要感染其他宿主——水鸟,但形态相似,主要是这些物种的雌性难以识别。在这里,我们描述了的形态学独特特征,并通过使用ITS1-5.8S-ITS2核糖体DNA进行分子分析来进行鉴定。对400份家禽胃肠道样本进行了为期2年的分析。使用光学显微镜和扫描电子显微镜(SEM)来确定的形态测量和特征形态特征。基于PCR的ITS1-5.8S-ITS2核糖体DNA测序用于分子表征,并通过MEGA 11进行系统发育分析。发现90份胃肠道样本被感染。雄虫具有发育良好的侧翼和尾翼,尾端有一个肛门吸盘、12对尾乳头和泄殖腔,从泄殖腔中伸出2根不等长的交合刺。雌虫有可见的阴道弯曲,阴门大约位于身体中部,尾端有一条长而尖的尾巴。序列分析显示,ITS1-5.8S-ITS2 rDNA的PCR产物大小为600 bp,在BLAST分析中与的ITS1-5.8S-ITS2序列显示出96-98%的相似性。我们的ITS序列与印度勒克瑙的相比,核苷酸差异为4.9%。系统发育分析清楚地将与该属的其他代表区分开来,并揭示了不同物种之间的关系。该研究揭示了在家禽中流行的感染状况,以及采取战略控制措施的必要性。研究结果表明,ITS rDNA将作为物种鉴定、诊断和系统分类的潜在分子标记。

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