实验动物与比较医学 ›› 2026, Vol. 46 ›› Issue (2): 222-230.DOI: 10.12300/j.issn.1674-5817.2025.087
王莹, 纪文韬, 徐少琼, 陈国元, 冯洁(
)(
), 吴宝金(
)(
)
收稿日期:2025-06-10
修回日期:2025-10-19
出版日期:2026-04-25
发布日期:2026-04-18
通讯作者:
作者简介:王 莹(1996—),女,硕士,助理工程师,研究方向:实验动物质量控制研究。E-mail:wangying2023@sibcb.ac.cn
WANG Ying, JI Wentao, XU Shaoqiong, CHEN Guoyuan, FENG Jie(
)(
), WU Baojin(
)(
)
Received:2025-06-10
Revised:2025-10-19
Published:2026-04-25
Online:2026-04-18
Contact:
FENG Jie (ORCID: 0009-0007-5364-9638), E-mail: fengjie@sibcb.ac.cn;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》和本屏障设施的《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的空气沉降菌、物料和人员手套等样本进行细菌监测,收集2020年1月到2024年12月的监测数据并进行整理统计,同时采用PCR和测序法对部分样品进行菌种鉴定。 结果 2020—2024年共采集样本7 898份,包括空气沉降菌样本3 175份、物料样本3 353份和手套样本1 370份。总体检测合格率为95.7%(7 559/7 898),其中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%(1 350/1 370)。5年间3类样本的合格率均在90%以上,且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,经分离培养获得细菌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9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细菌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。
中图分类号:
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的监测与分析[J]. 实验动物与比较医学, 2026, 46(2): 222-230. 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 Laboratory Animal Barrier Facilities[J]. Laboratory Animal and Comparative Medicine, 2026, 46(2): 222-230. DOI: 10.12300/j.issn.1674-5817.2025.087.
年度 Year | 空气沉降菌合格率 Air-settling bacteria compliance rate | 物料合格率 Material compliance rate | 手套合格率 Glove compliance rate | 总计 Total |
|---|---|---|---|---|
| 2020 | 99.7 (613/615) | 94.9 (616/649) | 99.3 (284/286) | 97.6 (1 513/1 550) |
| 2021 | 99.7 (634/636) | 95.5 (595/623) | 98.8 (319/323) | 97.9 (1 548/1 582) |
| 2022 | 94.5 (547/579) | 90.8 (546/601) | 99.5 (192/193) | 93.6 (1 285/1 373) |
| 2023 | 96.3 (649/674) | 92.8 (662/713) | 98.4 (242/246) | 95.1 (1 553/1 633) |
| 2024 | 95.5 (641/671) | 92.0 (706/767) | 97.2 (313/322) | 94.3 (1 660/1 760) |
| 总计 Total | 97.1 (3 084/3 175) | 93.2 (3 125/3 353) | 98.5 (1 350/1 370) | 95.7 (7 559/7 898) |
表1 2020—2024年屏障设施内样本的监测数量与合格率 (%(合格份数/样本数))
Table 1 Number and compliance rates of samples monitored in barrier facilities (2020—2024)
年度 Year | 空气沉降菌合格率 Air-settling bacteria compliance rate | 物料合格率 Material compliance rate | 手套合格率 Glove compliance rate | 总计 Total |
|---|---|---|---|---|
| 2020 | 99.7 (613/615) | 94.9 (616/649) | 99.3 (284/286) | 97.6 (1 513/1 550) |
| 2021 | 99.7 (634/636) | 95.5 (595/623) | 98.8 (319/323) | 97.9 (1 548/1 582) |
| 2022 | 94.5 (547/579) | 90.8 (546/601) | 99.5 (192/193) | 93.6 (1 285/1 373) |
| 2023 | 96.3 (649/674) | 92.8 (662/713) | 98.4 (242/246) | 95.1 (1 553/1 633) |
| 2024 | 95.5 (641/671) | 92.0 (706/767) | 97.2 (313/322) | 94.3 (1 660/1 760) |
| 总计 Total | 97.1 (3 084/3 175) | 93.2 (3 125/3 353) | 98.5 (1 350/1 370) | 95.7 (7 559/7 898) |
样本类别 Sample category | 第一季度 First quarter | 第二季度 Second quarter | 第三季度 Third quarter | 第四季度 Fourth quarter | 卡方值 χ2 value | P值 P value |
|---|---|---|---|---|---|---|
| 空气沉降菌Air-settling bacteria | 98.5 (724/735) | 98.8 (722/731) | 95.1 (796/837) | 96.6 (842/872) | 25.421 | <0.001 |
| 物料Materials | 94.2 (789/838) | 93.6 (690/737) | 90.8 (777/856) | 94.3 (869/922) | 10.986 | 0.012 |
| 手套Gloves | 98.3 (284/289) | 98.7 (308/312) | 98.4 (362/368) | 98.8 (396/401) | 0.416 | 0.940 |
表2 2020—2024年屏障设施内不同季度样本的监测数量和合格率 (%(合格份数/样本数))
Table 2 Number of monitored samples and compliance rates by quarter (2020—2024)
样本类别 Sample category | 第一季度 First quarter | 第二季度 Second quarter | 第三季度 Third quarter | 第四季度 Fourth quarter | 卡方值 χ2 value | P值 P value |
|---|---|---|---|---|---|---|
| 空气沉降菌Air-settling bacteria | 98.5 (724/735) | 98.8 (722/731) | 95.1 (796/837) | 96.6 (842/872) | 25.421 | <0.001 |
| 物料Materials | 94.2 (789/838) | 93.6 (690/737) | 90.8 (777/856) | 94.3 (869/922) | 10.986 | 0.012 |
| 手套Gloves | 98.3 (284/289) | 98.7 (308/312) | 98.4 (362/368) | 98.8 (396/401) | 0.416 | 0.940 |
环境样本来源 Source of environmental samples | 样本数量/份 Number of samples/n | 合格样本数量/份 Number of compliant samples/n | 合格率/% Compliance 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 |
表3 屏障设施内不同区域空气沉降菌的监测结果
Table 3 Monitoring results of air-settling bacteria in different areas of barrier facilities
环境样本来源 Source of environmental samples | 样本数量/份 Number of samples/n | 合格样本数量/份 Number of compliant samples/n | 合格率/% Compliance 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Gram-positive bacteria | 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Enterococcus | 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Gram-negative bacteria | 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Rodentibacter | 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 |
表4 屏障设施内不同区域空气沉降菌样本的细菌分离鉴定结果 (样本数/份)
Table 4 Results of isolation and identification of bacteria from air-settling bacteria samples in different areas of barrier facilities
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Gram-positive bacteria | 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Enterococcus | 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Gram-negative bacteria | 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Rodentibacter | 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 |
物料样本来源 Material sample sources | 样本数量/份 Number of samples/n | 合格样本数量/份 Number of compliant samples/n | 合格率/% Compliance rate/% |
|---|---|---|---|
| 料盒Feed box | 177 | 139 | 78.5 |
| 料铲Feed shovel | 177 | 151 | 85.3 |
| 饲料Feed | 167 | 152 | 91.0 |
超净工作台 Super clean bench | 1 539 | 1 417 | 92.1 |
| 铁架Iron shelf | 265 | 251 | 94.7 |
| 笼盖Cage cover | 325 | 316 | 97.2 |
| 笼盒Cage box | 457 | 453 | 99.1 |
| 垫料Bedding | 179 | 179 | 100.0 |
饮用水 Drinking water | 67 | 67 | 100.0 |
| 总计Total | 3 353 | 3 125 | 93.2 |
表5 屏障设施内不同物料样本的监测结果
Table 5 Monitoring results of different material samples in barrier facilities
物料样本来源 Material sample sources | 样本数量/份 Number of samples/n | 合格样本数量/份 Number of compliant samples/n | 合格率/% Compliance rate/% |
|---|---|---|---|
| 料盒Feed box | 177 | 139 | 78.5 |
| 料铲Feed shovel | 177 | 151 | 85.3 |
| 饲料Feed | 167 | 152 | 91.0 |
超净工作台 Super clean bench | 1 539 | 1 417 | 92.1 |
| 铁架Iron shelf | 265 | 251 | 94.7 |
| 笼盖Cage cover | 325 | 316 | 97.2 |
| 笼盒Cage box | 457 | 453 | 99.1 |
| 垫料Bedding | 179 | 179 | 100.0 |
饮用水 Drinking water | 67 | 67 | 100.0 |
| 总计Total | 3 353 | 3 125 | 93.2 |
细菌类别 Bacterial category | 料铲 Feed shovel | 铁架 Iron shelf | 笼盖 Cage cover | 料盒 Feed box | 超净工作台 Super clean bench | 总计 Total |
|---|---|---|---|---|---|---|
| 革兰氏阳性菌 Gram-positive bacteria | 12 | 7 | 8 | 8 | 17 | 52 |
| 葡萄球菌属 Staphylococcus | 11 | 5 | 7 | 8 | 17 | 48 |
| 木糖葡萄球菌 Staphylococcusxylosus | 5 | 2 | 0 | 4 | 9 | 20 |
| 尼泊尔葡萄球菌 Staphylococcus nepalensis | 2 | 1 | 4 | 2 | 5 | 14 |
| 缓慢葡萄球菌 Staphylococcuslentus | 1 | 2 | 2 | 2 | 2 | 9 |
| 科恩葡萄球菌 Staphylococcuscohnii | 3 | 0 | 0 | 0 | 0 | 3 |
| 松鼠葡萄球菌 Staphylococcussciuri | 0 | 0 | 1 | 0 | 0 | 1 |
| 腐生葡萄球菌 Staphylococcussaprophyticus | 0 | 0 | 0 | 0 | 1 | 1 |
| 气球菌属 Aerococcus | 1 | 2 | 1 | 0 | 0 | 4 |
| 绿色气球菌 Aerococcusviridans | 1 | 2 | 1 | 0 | 0 | 4 |
表6 屏障设施内不同物料样本的细菌分离鉴定结果 (样本数/份)
Table 6 Results of isolation and identification of bacteria from different material samples in barrier facilities
细菌类别 Bacterial category | 料铲 Feed shovel | 铁架 Iron shelf | 笼盖 Cage cover | 料盒 Feed box | 超净工作台 Super clean bench | 总计 Total |
|---|---|---|---|---|---|---|
| 革兰氏阳性菌 Gram-positive bacteria | 12 | 7 | 8 | 8 | 17 | 52 |
| 葡萄球菌属 Staphylococcus | 11 | 5 | 7 | 8 | 17 | 48 |
| 木糖葡萄球菌 Staphylococcusxylosus | 5 | 2 | 0 | 4 | 9 | 20 |
| 尼泊尔葡萄球菌 Staphylococcus nepalensis | 2 | 1 | 4 | 2 | 5 | 14 |
| 缓慢葡萄球菌 Staphylococcuslentus | 1 | 2 | 2 | 2 | 2 | 9 |
| 科恩葡萄球菌 Staphylococcuscohnii | 3 | 0 | 0 | 0 | 0 | 3 |
| 松鼠葡萄球菌 Staphylococcussciuri | 0 | 0 | 1 | 0 | 0 | 1 |
| 腐生葡萄球菌 Staphylococcussaprophyticus | 0 | 0 | 0 | 0 | 1 | 1 |
| 气球菌属 Aerococcus | 1 | 2 | 1 | 0 | 0 | 4 |
| 绿色气球菌 Aerococcusviridans | 1 | 2 | 1 | 0 | 0 | 4 |
细菌类别 Bacterial category | 样本数量/份 Number of samples/n |
|---|---|
| 革兰氏阳性菌Gram-positive bacteria | 12 |
| 葡萄球菌属Staphylococcus | 6 |
| 木糖葡萄球菌Staphylococcusxylosus | 2 |
| 表皮葡萄球菌Staphylococcusepidermidis | 1 |
| 缓慢葡萄球菌Staphylococcuslentus | 1 |
| 尼泊尔葡萄球菌Staphylococcusnepalensis | 1 |
| 溶血性葡萄球菌Staphylococcushaemolyticus | 1 |
| 芽孢杆菌属Bacillus | 2 |
| 高地芽孢杆菌Bacillusaltitudinis | 1 |
| 蜡样芽孢杆菌Bacilluscereus | 1 |
| 类芽孢杆菌属Paenibacillus | 2 |
| 解木聚糖类芽孢杆菌Paenibacillusxylanilyticus | 1 |
| 解葡聚糖类芽孢杆菌Paenibacillus glucanolyticus | 1 |
| 大洋芽孢杆菌属Oceanobacillus | 1 |
| 黄豆大洋芽孢杆菌Oceanobacillus sojae | 1 |
| 气球菌属Aerococcus | 1 |
| 绿色气球菌Aerococcusviridans | 1 |
表7 屏障设施内手套样本的细菌分离鉴定结果
Table 7 Results of isolation and identification of bacteria from glove samples in barrier facilities
细菌类别 Bacterial category | 样本数量/份 Number of samples/n |
|---|---|
| 革兰氏阳性菌Gram-positive bacteria | 12 |
| 葡萄球菌属Staphylococcus | 6 |
| 木糖葡萄球菌Staphylococcusxylosus | 2 |
| 表皮葡萄球菌Staphylococcusepidermidis | 1 |
| 缓慢葡萄球菌Staphylococcuslentus | 1 |
| 尼泊尔葡萄球菌Staphylococcusnepalensis | 1 |
| 溶血性葡萄球菌Staphylococcushaemolyticus | 1 |
| 芽孢杆菌属Bacillus | 2 |
| 高地芽孢杆菌Bacillusaltitudinis | 1 |
| 蜡样芽孢杆菌Bacilluscereus | 1 |
| 类芽孢杆菌属Paenibacillus | 2 |
| 解木聚糖类芽孢杆菌Paenibacillusxylanilyticus | 1 |
| 解葡聚糖类芽孢杆菌Paenibacillus glucanolyticus | 1 |
| 大洋芽孢杆菌属Oceanobacillus | 1 |
| 黄豆大洋芽孢杆菌Oceanobacillus sojae | 1 |
| 气球菌属Aerococcus | 1 |
| 绿色气球菌Aerococcusviridans | 1 |
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