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 ™ 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.

中文翻译：

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高通量测序方法揭示了熊熊ter中的寄生虫种类繁多（Linnaeus，1758）

评估寄生虫多样性的程度需要使用适当的分子工具，尤其是考虑到越来越多的证据表明多种寄生虫同时存在。在这里，我们在之前的研究中比较了12份经PCR鉴定为锥虫（Leishmaniinae）阳性的Bombus terrestris标本中的新一代测序技术（Ion PGM™System）的性能。还使用经典方案（特异性PCR扩增或使用大范围引物加Sanger测序的扩增）和离子PGM测序对这些大黄蜂进行了Nosematidae和Neogregarinorida寄生虫的筛查。后者揭示了个体内的寄生虫多样性更高，尤其是在利什曼虫科中（这些寄生虫是Lotmaria passim，Critidia mellificae和Crithidia bombi）以及这些寄主中未曾报道过的分类单元的发生：Cristidia acanthocephali和一种新的新gregarinorida种。此外，不同引物组产生的互补结果凸显了使用多种标记物以最大程度减少某些目标生物被忽视的机会的便利性。总之，深度测序方法提供了比通常的鉴定方法更全面的方法来研究寄生虫多样性，并提供了新的见解，其对于大黄蜂健康的重要性应进一步分析。