Eupolyphaga sinensis Walker is a valuable traditional Chinese animal medicine first recorded in Shennong Bencao. Previous research has shown that E. sinensis is easily contaminated by aflatoxins (AFs), which are highly toxic mycotoxins, during harvest, storage, and transport, thereby posing a considerable threat to consumer health. Most often, these AFs are produced by Aspergillus species. In this study, we contrast the traditional culture-based dilution plating method to the high-throughput sequencing (HTS) technology for fungal identification in TCM E. sinensis. Both of the methods used internal transcribed spacer 1 (ITS1) and calmodulin (CaM) sequencing for fungal molecular identification. The new CaM primer we designed in the study is suitable for MiSeq PE300 sequencing used for identification of Aspergillus species in community DNA samples. More fungal species were found in the E. sinensis samples based on HTS than those found using the culture-based dilution plating method. Overall, combining the sequencing power of ITS1 and CaM is an effective method for the detection and monitoring of potential toxigenic Aspergillus species in E. sinensis. In conclusion, HTS can be used to obtain a large amount of sequencing data about fungi contaminating animal medicine, allowing earlier detection of potential toxigenic fungi and ensuring the efficient production and safety of E. sinensis.

中文翻译：

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基于 ITS1 和 CaM 高通量测序的土鳖虫分枝杆菌和曲霉属的分子特征

土鳖虫是一种名贵的中药动物药，最早记载于神农本草。此前的研究表明，黄曲霉在收获、储存和运输过程中很容易被黄曲霉毒素（AFs）污染，黄曲霉毒素是一种剧毒的霉菌毒素，对消费者的健康构成了相当大的威胁。大多数情况下，这些 AF 是由曲霉属产生的。在这项研究中，我们将传统的基于培养的稀释接种方法与高通量测序 (HTS) 技术进行了对比，用于中药 E. sinensis 的真菌鉴定。这两种方法都使用内部转录间隔区 1 (ITS1) 和钙调蛋白 (CaM) 测序进行真菌分子鉴定。我们在研究中设计的新型 CaM 引物适用于 MiSeq PE300 测序，用于鉴定群落 DNA 样本中的曲霉属物种。在基于 HTS 的 E. sinensis 样品中发现的真菌物种比使用基于培养的稀释电镀方法发现的真菌物种多。总的来说，结合 ITS1 和 CaM 的测序能力是检测和监测 E. sinensis 中潜在产毒曲霉物种的有效方法。综上所述，HTS可用于获取大量污染动物药的真菌的测序数据，可以及早发现潜在的产毒真菌，确保中华绒螯蟹的高效生产和安全。