一批微生物超标面膜中日勾维多细菌源菌的分离鉴定及毒力与耐药性分析

Isolation, identification, and virulence and drug resistance examination of Pluralibacter gergoviae from a batch of facial masks with microorganisms out of limits

摘要:

【背景】 近年来，由不可接受微生物污染引发的化妆品召回案件层出不穷，现有法规《化妆品安全技术规范》(2015年版)中微生物检测指标不完善，难以满足化妆品产品质量控制和评价的需要。【目的】 对2022年化妆品国家级监督抽检中一批微生物超标面膜中污染菌进行分离鉴定，并探讨采用多种手段进行鉴定的准确性，判断是否为不可接受微生物，同时，针对污染菌进一步开展安全性评价。【方法】 参照《化妆品安全技术规范》(2015年版)对微生物超标面膜中污染菌进行分离纯化后，分别采用VITEK 2 Compact全自动生化鉴定系统、基质辅助激光解吸电离飞行时间质谱(matrix assisted laser desorption ionization-time of flight mass spectrometry, MALDI-TOF MS)和16S rRNA基因测序技术对污染菌进行鉴定，并进一步开展毒力基因、耐药基因安全性评价分析。【结果】 3种技术均可鉴定出面膜中污染菌为不可接受微生物——日勾维多细菌源菌。高通量测序发现该菌携带52种毒力基因，预测含其他毒力相关基因429种，提示该菌具有极大的安全隐患。同时检测到该菌携带24种耐药基因，可能对青霉素、头孢菌素、万古霉素、四环素和氨基糖苷类等24种(类)抗生素耐药，检测到多重耐药基因10种，耐药机理多与产生外排泵有关。提示该菌存在一定的多重耐药安全风险。【结论】 本研究从一批微生物超标面膜中分离到日勾维多细菌源菌，为不可接受微生物。该菌具有极大的产毒能力，且具有一定的多重耐药安全隐患，应考虑经风险评估后将该菌纳入微生物检测指标。

关键词:不可接受微生物;微生物超标面膜;日勾维多细菌源菌;分离鉴定;毒力基因;耐药基因

Abstract:

[Background] In recent years, cosmetic recall cases caused by contamination with objectionable microorganisms have kept emerging. The microbial detection indicators in the Cosmetic Safety Technical Specifications (2015 Edition) are not perfect and difficult to meet the needs of quality control and evaluation of cosmetics products. [Objective] To isolate and identify the contaminant microorganisms in a batch of facial masks with microorganisms out of limits in the national sampling inspection of cosmetics in 2022, explore the performance of different identification methods, determine whether the contaminant microorganisms are objectionable microorganisms or not, and evaluate the safety of the contaminant microorganisms.[Methods] According to the Cosmetic Safety Technical Specifications (2015 Edition), the contaminant microorganisms in the facial masks were isolated and identified by VITEK 2 system, matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), and 16S rRNA gene sequencing. Furthermore, the safety evaluation and detection of virulence genes and drug resistance genes were carried out for the contaminant microorganisms. [Results] The results obtained with the three identification techniques all showed that the contaminant microorganism in the facial masks was an objectionable microorganism, Pluralibacter gergoviae. High-throughput sequencing detected 52 virulence genes and predicted 429 other virulence-related genes, which suggested that this bacterium had great safety risks. At the same time, 24 drug resistance genes were detected, suggesting that the bacterium may be resistant to 24 antibiotics such as penicillin, cephalosporin, vancomycin, tetracycline, and aminoglycosides. Moreover, 10 multi-drug resistance genes were detected, and the resistance mechanism was associated with the production of efflux pumps. It is suggested that the bacterium have a certain risk of multi-drug resistance. [Conclusion] In this study, an objectionable microorganism P. gergoviae was isolated from a batch of facial masks with microorganisms out of limit. This bacterium is capable of producing toxins and has a potential safety risk of multidrug resistance. It should be considered to include this bacterium into the microbial detection after risk assessment.

Key words:objectionable microorganisms;facial mask with microorganisms out of limits;Pluralibacter gergoviae;isolation and identification;virulence genes;resistance genes