实验动物屏障设施环境微生物污染情况监测与分析

doi:10.12300/j.issn.1674-5817.2025.087

摘要/Abstract

摘要：

目的了解实验动物屏障设施内微生物污染情况与分布特征，为屏障设施环境质量控制提供科学依据。方法根据国家标准《实验动物环境及设施》和屏障设施的《标准操作规程》，对中国科学院分子细胞科学卓越创新中心动物实验技术平台单走廊屏障设施内空气沉降菌、物料和人员手套等样本进行细菌监测，将2020~2024年的监测数据进行整理统计，并利用PCR和测序法对部分样品进行了菌种鉴定。结果 2020~2024年间共采集样本7 898份，包括空气沉降菌样本3 175份、物料样本3 353份以及手套样本1 370份。总体合格率为95.7%（7 559/7 898），空气沉降菌合格率为97.1%（3 084/3 175）、物料合格率为93.2%（3 125/3 353）、人员手套合格率为98.5%（1350/1370）。5年间监测3类样本的合格率均在90%以上；不同季度间空气沉降菌和物料样本合格率差异有统计学意义（P＜0.05）。对2024年1~3月采集的样本进一步研究，分离培养获得细菌190株，其中空气沉降菌126株、物料来源菌52株、人员手套来源菌12株。经PCR扩增和测序鉴定，190株细菌分属7个菌属20个菌种，革兰氏阳性菌占多数，以葡萄球菌属为主，占比77.9%（148/190）。结论屏障设施内空气、物料和人员手套等携带的微生物以革兰氏阳性菌为主。通过定期对屏障设施内空气沉降菌、物料和人员手套进行监测，可及时发现并控制饲养管理和设施运行过程中的潜在风险，对维持屏障设施系统良好运行、保障动物实验质量具有重要意义。

关键词: 实验动物, 屏障设施, 质量控制, 细菌鉴定

Abstract:

Objective To understand the microbial contamination and distribution characteristics in barrier facility of laboratory animal, and to provide scientific basis for environmental quality control of housing facilities. Methods According to national standard"Laboratory animal-Environment and housing facilities" and the "Standard Operating Procedures" of the barrier facility, bacterial monitoring was carried out on samples of air-settling bacteria, materials and personnel gloves in the single-channel barrier facility of laboratory animal of the Animal Core Facility of the Center for Excellence in Molecular Cell Science of the Chinese Academy of Sciences(CEMCS), Monitoring data from 2020 to 2024 were collated and statistically analyzed, with species identification performed on selected samples using PCR and sequencing methods. Results Between 2020 and 2024, a total of 7 898 samples were collected, including 3 175 samples of airborne bacteria, 3 353 samples of materials, and 1370 samples of gloves. The overall pass rate is 95.7% (7 559/7 898), the pass rate for air settling bacteria is 97.1% (3 084/3 175), the pass rate for materials is 93.2% (3 125/3 353), and the pass rate for personnel gloves is 98.5% (1 350/1 370). Over a five-year period, the compliance rate for all three sample categories remained above 90%. Statistically significant differences (P<0.05) were observed in compliance rates for airborne microbial samples and material samples across different quarters.The samples collected from January-March 2024 were further studied and 190 strains of bacteria were isolated and cultured and obtained, of which 126 strains were air deposition bacteria, 52 strains were material source bacteria and 12 strains were personnel glove source bacteria. After PCR amplification and sequencing identification, 190 bacteria were classified into 7 genera and 20 species, with Gram positive bacteria accounting for the majority, mainly Staphylococcus, accounting for 77.9% (148/190). Conclusion Microorganisms carried by air, materials and personnel gloves inside the barrier facility are mainly Gram-positive bacteria. Regular monitoring of airborne bacteria, materials and personnel gloves in the barrier facility can detect and control potential risks in the process of feeding management and facility operation, which is of great significance in maintaining the good operation of the barrier facility system and guaranteeing the quality of animal experiments.

Key words: Laboratory animals, Barrier facility, Quality control, Bacterial identification

中图分类号:

Q95-33

王莹,纪文韬,徐少琼,等.实验动物屏障设施环境微生物污染情况监测与分析[J]. 实验动物与比较医学.. DOI: 10.12300/j.issn.1674-5817.2025.087.

WANG Ying,JI Wentao,XU Shaoqiong,et al. Monitoring and Analysis of Environmental Microbial Contamination in Barrier Facility of Laboratory Animal[J]. Laboratory Animal and Comparative Medicine. DOI: 10.12300/j.issn.1674-5817.2025.087.

图/表 6

参考文献 23

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年度 Year	空气沉降菌合格率 Falling bacteria pass rate	物料合格率 Material pass rate	手套合格率 Gloves pass rate	总计 Total
2020	99.7%(613/615)	94.9%(616/649)	99.3%(284/286)	97.6%(1513/1550)
2021	99.7%(634/636)	95.5%(595/623)	98.8%(319/323)	97.9%(1548/1582)
2022	94.5%(547/579)	90.8%(546/601)	99.5%(192/193)	93.6%(1285/1373)
2023	96.3%(649/674)	92.8%(662/713)	98.4%(242/246)	95.1%(1553/1633)
2024	95.5%(641/671)	92.0%(706/767)	97.2%(313/322)	94.3%(1660/1760)
总计Total	97.1%(3084/3175)	93.2%(3125/3353)	98.5%(1350/1370)	95.7%(7559/7898)

年度 Year	空气沉降菌合格率 Falling bacteria pass rate	物料合格率 Material pass rate	手套合格率 Gloves pass rate	总计 Total
2020	99.7%(613/615)	94.9%(616/649)	99.3%(284/286)	97.6%(1513/1550)
2021	99.7%(634/636)	95.5%(595/623)	98.8%(319/323)	97.9%(1548/1582)
2022	94.5%(547/579)	90.8%(546/601)	99.5%(192/193)	93.6%(1285/1373)
2023	96.3%(649/674)	92.8%(662/713)	98.4%(242/246)	95.1%(1553/1633)
2024	95.5%(641/671)	92.0%(706/767)	97.2%(313/322)	94.3%(1660/1760)
总计Total	97.1%(3084/3175)	93.2%(3125/3353)	98.5%(1350/1370)	95.7%(7559/7898)

环境样本来源 Sources of environmental samples	样本数量 Sample size	合格份数 Number of qualifications	合格率Satisfactory rate
饲养间 Breeding room	1 718	1 636	95.2%
风淋间 Air shower room	183	179	97.8%
缓冲间 Buffer room	183	180	98.4%
走廊 Corridor	306	304	99.3%
隔离检疫间 Quarantine room	183	183	100.0%
实验操作间 Laboratory operating room	351	351	100.0%
消毒后室 Sterilization room	251	251	100.0%
总计 Total	3 175	3 084	97.1%

环境样本来源 Sources of environmental samples	样本数量 Sample size	合格份数 Number of qualifications	合格率Satisfactory rate
饲养间 Breeding room	1 718	1 636	95.2%
风淋间 Air shower room	183	179	97.8%
缓冲间 Buffer room	183	180	98.4%
走廊 Corridor	306	304	99.3%
隔离检疫间 Quarantine room	183	183	100.0%
实验操作间 Laboratory operating room	351	351	100.0%
消毒后室 Sterilization room	251	251	100.0%
总计 Total	3 175	3 084	97.1%

细菌类别 Bacterial category	饲养间Breeding room	隔离检疫间Quarantine room	实验操作间Laboratory operating room	消毒后室Sterilization room	走廊Corridor	风淋间 Air shower room	缓冲间Buffer room	总计Total
革兰氏阳性菌G+	99	0	2	1	13	3	6	124
葡萄球菌属 Staphylococcus	75	0	0	0	12	2	5	94
木糖葡萄球菌 Staphylococcusxylosus	48	0	0	0	10	0	2	60
缓慢葡萄球菌 Staphylococcuslentus	18	0	0	0	1	0	1	20
松鼠葡萄球菌 Staphylococcussciuri	4	0	0	0	1	0	1	6
尼泊尔葡萄球菌 Staphylococcus nepalensis	3	0	0	0	0	0	0	3
腐生葡萄球菌 Staphylococcussaprophyticus	2	0	0	0	0	0	1	3
表皮葡萄球菌 Staphylococcusepidermidis	0	0	0	0	0	2	0	2
气球菌属 Aerococcus	19	0	0	0	1	0	1	21
绿色气球菌 Aerococcusviridans	17	0	0	0	1	0	1	19
脲气球菌 Aerococcusurinae	2	0	0	0	0	0	0	2
肠球菌属 EnterococcusThiercelinandJouhaud	4	0	2	0	0	0	0	6
粪肠球菌 Enterococcusfaecalis	4	0	2	0	0	0	0	6
芽孢杆菌属 Bacillus	1	0	0	1	0	0	0	2
蜡样芽孢杆菌 Bacilluscereus	1	0	0	0	0	0	0	1
尼氏芽孢杆菌 Bacillusnealsonii	0	0	0	1	0	0	0	1
微球菌属 Micrococcus	0	0	0	0	0	1	0	1
藤黄微球菌 Micrococcusluteus	0	0	0	0	0	1	0	1
革兰氏阴性菌G-	1	1	0	0	0	0	0	2
不动杆菌属 Acinetobacter	0	1	0	0	0	0	0	1
申氏不动杆菌 Acinetobacterschindleri	0	1	0	0	0	0	0	1
啮齿杆菌属Rodentibactergenus	1	0	0	0	0	0	0	1
嗜肺啮齿杆菌 Rodentibacter pneumotropicus	1	0	0	0	0	0	0	1
总计 Total	100	1	2	1	13	3	6	126