Dong Lin, Zhang Zilong, Zhu Biyun, Li Shenwei, He Yan, Lou Yating, Li Ping, Zheng Huajun, Tian Zhengan, Ma Xia
School of Perfume and Aroma Technology, Shanghai Institute of Technology, Shanghai, China.
Shanghai International Travel Healthcare Center, Shanghai, China.
Front Med (Lausanne). 2022 Sep 21;9:963988. doi: 10.3389/fmed.2022.963988. eCollection 2022.
Microbial inoculants are widely used in wastewater treatment, soil remediation, and biological control. Safety and compliance for active constituents are considered to be the most important measures of imported microbial inoculants. Microbial inoculants composition was commonly identified by phenotypic culture, which is time-consuming and labor intense with occasionally false negative results provided, and can only be tested for specific species. High-throughput sequencing (HTS), known for its non-targeted detection of unknown species composition in samples, is suitable for composition consistency identification and biosafety analysis of imported microbial inoculants. In this study, the application of HTS for microflora distribution and resistance gene was verified in microbial inoculants for environmental protection and then applicated in imported microbial inoculants. Both Illumina- and Nanopore-based HTS methods identified the same dominant bacterial species successfully in the imported microbial inoculants. The main component of bacterial species was , and , and further confirmed with traditional methods. The antibiotic resistance genes were detected in all samples. Our results indicated that HTS processes the application potential to identify the active ingredients of microbial inoculants. Therefore, rapid and accurate identification of the microbial compositions in microbial formulation products is of high importance for port biosafety supervision.
微生物接种剂广泛应用于废水处理、土壤修复和生物防治。活性成分的安全性和合规性被认为是进口微生物接种剂最重要的指标。微生物接种剂的成分通常通过表型培养来鉴定，这种方法既耗时又费力，偶尔还会出现假阴性结果，而且只能针对特定物种进行检测。高通量测序（HTS）以其对样品中未知物种组成的非靶向检测而闻名，适用于进口微生物接种剂的成分一致性鉴定和生物安全性分析。在本研究中，HTS在环保型微生物接种剂中对微生物群落分布和抗性基因的应用得到了验证，随后应用于进口微生物接种剂。基于Illumina和Nanopore的HTS方法均成功鉴定出进口微生物接种剂中相同的优势细菌物种。细菌物种的主要成分是 ， ，并通过传统方法进一步确认。所有样品中均检测到抗生素抗性基因 。我们的结果表明，HTS在鉴定微生物接种剂活性成分方面具有应用潜力。因此，快速准确地鉴定微生物制剂产品中的微生物组成对于口岸生物安全监管至关重要。
