Mussels’ aquaculture is one of the most important cultures in Europe, with a high level of self-sufficiency. In Spain, the main cultivated species is , whose production takes place primarily in Galicia. In that region, a Protected Designation of Origin certification exists to certify the high quality of the product and its sustainability. This species is also present in other regions making the control of its traceability complicated. In order to distinguish Galician mussels from the others, the eukaryotes present in its digestive gland could be a useful tool. However, to use them as biomarkers, having an adapted protocol to recover the diversity present in the digestive gland is essential. In this work, we have selected a new protocol to optimize the study of the diversity of eukaryotes associated with the digestive gland of . This protocol is based on a combination of a pre-treatment and a commercial DNA extraction kit, the amplification of DNA with two newly designed primer sets to avoid the DNA from the mussel, and the use of next-generation sequencing. The results showed a significant reduction of mussel DNA amplification and enough DNA quality for the subsequent sequencing, increasing the number of sequences recovered from other eukaryotes. This new protocol offers then a good chance to determine the diversity of eukaryotes present in the digestive gland of , and it can be used in future studies to examinate the seasonal variation of the present species and to evaluate its potential as geographical traceability tool.

中文翻译：

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用于研究地中海贻贝消化腺中存在的真核生物的新宏基因组程序

贻贝水产养殖是欧洲最重要的文化之一，具有高度的自给自足能力。在西班牙，主要栽培品种是 ，其生产主要在加利西亚。在该地区，存在受保护的原产地命名认证，以证明产品的高质量及其可持续性。该物种也存在于其他地区，这使得其可追溯性的控制变得复杂。为了将加利西亚贻贝与其他贻贝区分开来，其消化腺中存在的真核生物可能是一个有用的工具。然而，要将它们用作生物标志物，必须采用合适的方案来恢复消化腺中存在的多样性。在这项工作中，我们选择了一个新的方案来优化与消化腺相关的真核生物多样性的研究。该方案基于预处理和商业 DNA 提取试剂盒的组合、使用两个新设计的引物组扩增 DNA 以避免贻贝中的 DNA，以及使用下一代测序。结果显示，贻贝 DNA 扩增显着减少，并且 DNA 质量足以满足后续测序的要求，从而增加了从其他真核生物中回收的序列数量。这个新协议提供了一个很好的机会来确定消化腺中存在的真核生物的多样性，并且它可以在未来的研究中用于检查现有物种的季节性变化并评估其作为地理可追溯工具的潜力。