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OTUs 和 ASVs 使用定制的丰度过滤器从虾微生物群 16S 图谱中产生可比的分类和多样性。

OTUs and ASVs Produce Comparable Taxonomic and Diversity from Shrimp Microbiota 16S Profiles Using Tailored Abundance Filters.

机构信息

Departamento de Microbiología Molecular, Instituto de Biotecnología (IBT), Universidad Nacional, Autónoma de México (UNAM) Avenida Universidad #2001, Colonia Chamilpa, Cuernavaca, Morelos 62210, Mexico.

Departamento de Ciencias Químico Biológicas, Universidad de Sonora (UNISON), Blvd., Rosales y Luis, Encinas, Hermosillo, Sonora 83000, Mexico.

出版信息

Genes (Basel). 2021 Apr 13;12(4):564. doi: 10.3390/genes12040564.

Abstract

The interplay between shrimp immune system, its environment, and microbiota contributes to the organism's homeostasis and optimal production. The metagenomic composition is typically studied using 16S rDNA profiling by clustering amplicon sequences into operational taxonomic units (OTUs) and, more recently, amplicon sequence variants (ASVs). Establish the compatibility of the taxonomy, α, and β diversity described by both methods is necessary to compare past and future shrimp microbiota studies. Here, we used identical sequences to survey the V3 16S hypervariable-region using 97% and 99% OTUs and ASVs to assess the hepatopancreas and intestine microbiota of from two ponds under standardized rearing conditions. We found that applying filters to retain clusters >0.1% of the total abundance per sample enabled a consistent taxonomy comparison while preserving >94% of the total reads. The three sets turned comparable at the family level, whereas the 97% identity OTU set produced divergent genus and species profiles. Interestingly, the detection of organ and pond variations was robust to the clustering method's choice, producing comparable α and β-diversity profiles. For comparisons on shrimp microbiota between past and future studies, we strongly recommend that ASVs be compared at the family level to 97% identity OTUs or use 99% identity OTUs, both using tailored frequency filters.

摘要

虾免疫系统、其环境和微生物群之间的相互作用有助于维持生物体的内稳态和最佳生产。通常使用 16S rDNA 分析通过将扩增子序列聚类为操作分类单元 (OTUs) 来研究宏基因组组成,最近则使用扩增子序列变体 (ASVs)。为了比较过去和未来的虾微生物组研究,有必要确定这两种方法描述的分类学、α 和β多样性的兼容性。在这里,我们使用相同的序列,使用 97%和 99%的 OTUs 和 ASVs 对来自两个标准化养殖条件下池塘的虾的肝胰腺和肠道微生物群进行调查,使用 V3 16S 高变区 97% 的 OTUs 和 ASVs。我们发现,应用过滤器保留每个样本总丰度>0.1%的聚类可以进行一致的分类学比较,同时保留>94%的总读数。在科水平上,这三组变得可比,而 97%同一性 OTU 集产生了不同的属和种分布。有趣的是,器官和池塘变化的检测对聚类方法的选择具有鲁棒性,产生可比的α和β多样性分布。对于过去和未来虾微生物组研究之间的比较,我们强烈建议在科水平上将 ASVs 与 97%同一性 OTUs 或使用 99%同一性 OTUs 进行比较,两者都使用定制的频率过滤器。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ab4b/8070570/9cdffb419f0b/genes-12-00564-g001.jpg

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