伊朗自然感染绵羊皱胃中处于发育停滞期的捻转血矛线虫幼虫的首次分子特征和季节性分析。

First Molecular Characterization and Seasonality of Larvae of Trichostrongylid Nematodes in Arrested Development in the Abomasum of Iranian Naturally Infected Sheep.

机构信息

Department of Pathobiology, Faculty of Veterinary Medicine, Urmia University, Nazlu Campus, POBox: 57153-1177, Urmia, Iran.

Department of Microbiology, Faculty of Veterinary Sciences, Ilam University, Ilam, Iran.

出版信息

Acta Parasitol. 2021 Mar;66(1):193-198. doi: 10.1007/s11686-020-00274-3. Epub 2020 Sep 16.

DOI:10.1007/s11686-020-00274-3

PMID:32939698

Abstract

PURPOSE

The aim of this study was to determine molecularly the species of larvae of trichostrongylid nematodes in arrested development in the abomasum of sheep from Iran and their seasonality.

METHODS

A total of 240 sheep abomasa were randomly collected at different seasons from Ilam abattoir in 2017-2018. The adult nematodes were isolated and identified. The mucosa of the abomasum was removed and digested using a pepsin solution. DNA from different parts of the abomasum was extracted using a SinaPureTM kit. Polymerase chain reaction (PCR) amplification was performed; the ITS2-rRNA gene was amplified using specific primers. Amplified PCR products were purified and sequenced by Bioneer Corporation (South Korea). The obtained ITS2 sequences were aligned using BLAST analysis and submitted to GenBank.

RESULTS

Morphologically, five species of adult nematodes of four genera including Haemonchus contortus, Marshallagia marshalli, Marshallagia occidentalis, Parabronema skrjabini, and Teladorsagia circumcincta were identified. Marshallagia marshalli (65.4%, 120.2 ± 28.5) had the highest prevalence during autumn. Molecularly, arrested development of larvae of M. marshalli and T. circumcincta were demonstrated in mucosa of the abomasum in autumn and early winter using PCR of the ITS-2 region of rRNA.

CONCLUSION

The results evidenced the first molecular characterization of the larvae of trichostrongylid nematodes from abomasum of sheep from Iran.

摘要

目的

本研究旨在确定伊朗绵羊皱胃中处于发育停滞状态的捻转血矛线虫幼虫的种属，并分析其季节性。

方法

2017-2018 年，我们从伊拉姆屠宰场随机采集了 240 只绵羊的皱胃。分离并鉴定了成虫。用胃蛋白酶溶液消化皱胃黏膜。使用 SinaPureTM 试剂盒从皱胃的不同部位提取 DNA。采用特定引物进行聚合酶链反应（PCR）扩增；用 ITS2-rRNA 基因扩增。纯化扩增的 PCR 产物，并由 Bioneer 公司（韩国）进行测序。所得 ITS2 序列使用 BLAST 分析进行比对，并提交至 GenBank。

结果

形态学上，共鉴定出 4 个属的 5 种成虫线虫，包括捻转血矛线虫、马歇尔线虫、 occidentalis 马歇尔线虫、Skrjabinema 副柔线虫和 circumcincta 有齿食道口线虫。秋季，M. marshalli（65.4%，120.2±28.5）的检出率最高。分子检测结果显示，在秋季和初冬，利用 rRNA 的 ITS-2 区 PCR，在绵羊皱胃黏膜中可以检测到 M. marshalli 和 T. circumcincta 幼虫的发育停滞。