Wide diversity of parasites in Bombus terrestris (Linnaeus, 1758) revealed by a high-throughput sequencing approach
Wide diversity of parasites in Bombus terrestris (Linnaeus, 1758) revealed by a high-throughput sequencing approach
复制标题
DOI：
10.1111/1462-2920.15336
复制
发表时间：
2020-12-04
影响因子：
5.1
通讯作者：
Maside, Xulio
中科院分区：
文献类型：
--
作者：
Bartolome, Carolina;Jabal-Uriel, Clara;Maside, Xulio
评估寄生虫多样性的程度需要应用适当的分子工具，特别是考虑到越来越多的证据表明多种寄生虫共存。在这里，我们比较了性能的下一代测序技术（离子PGM（TM）系统）在12熊蜂terrestris标本，PCR鉴定为阳性锥虫（利什曼虫亚科）在以前的研究。这些熊蜂也筛选微孢子虫和Neogelastomerinorida寄生虫的发生，使用两个经典的协议（无论是特异性PCR扩增或扩增与广泛的引物加桑格测序）和离子PGM测序。后者揭示了更高的寄生虫多样性的个人，特别是利什曼虫亚科（这是目前作为一个组合的Lotmaria百香果，Crithidia mellificae和Crithidia bombi），并发生从未报道过在这些主机的分类群：Crithidia acanthocephali和一个新的neogalphinorida物种。此外，由不同引物组产生的互补结果突出了使用多个标记的便利性，以最大限度地减少一些靶生物被忽视的机会。总而言之，深度测序方法提供了一种比通常的鉴定方法更全面的方法来调查寄生虫多样性，并提供了新的见解，其对大黄蜂健康的重要性应进一步分析。
Assessing the extent of parasite diversity requires the application of appropriate molecular tools, especially given the growing evidence of multiple parasite co-occurrence. Here, we compared the performance of a next-generation sequencing technology (Ion PGM (TM) System) in 12 Bombus terrestris specimens that were PCR-identified as positive for trypanosomatids (Leishmaniinae) in a previous study. These bumblebees were also screened for the occurrence of Nosematidae and Neogregarinorida parasites using both classical protocols (either specific PCR amplification or amplification with broad-range primers plus Sanger sequencing) and Ion PGM sequencing. The latter revealed higher parasite diversity within individuals, especially among Leishmaniinae (which were present as a combination of Lotmaria passim, Crithidia mellificae and Crithidia bombi), and the occurrence of taxa never reported in these hosts: Crithidia acanthocephali and a novel neogregarinorida species. Furthermore, the complementary results produced by the different sets of primers highlighted the convenience of using multiple markers to minimize the chance of some target organisms going unnoticed. Altogether, the deep sequencing methodology offered a more comprehensive way to investigate parasite diversity than the usual identification methods and provided new insights whose importance for bumblebee health should be further analysed.