Lai Peggy S, Allen Joseph G, Hutchinson Diane S, Ajami Nadim J, Petrosino Joseph F, Winters Thomas, Hug Christopher, Wartenberg Gary R, Vallarino Jose, Christiani David C
Division of Pulmonary and Critical Care, Massachusetts General Hospital, Boston, MA, United States of America.
Department of Environmental Health, Harvard T.H. Chan School of Public Health, Boston, MA, United States of America.
PLoS One. 2017 Jul 13;12(7):e0180969. doi: 10.1371/journal.pone.0180969. eCollection 2017.
To characterize the microbial environment of workers in academic mouse research facilities using endotoxin, 16S qPCR, and 16S amplicon sequencing. To determine whether the work microbiome contributes to the human microbiome of workers.
We performed area air sampling from the animal rooms, dirty, middle, and setup cage wash locations in four academic mouse research facilities. 10 workers in the dirty cage wash area underwent personal air sampling as well as repeated collection of nasal, oral, and skin samples before and after the work shift. Environmental samples underwent measurement of endotoxin, mouse allergen, bacteria copy number via 16S qPCR, and microbial identification via 16S rDNA sequencing. 16S rDNA sequencing was also performed on human samples before and after the work shift. SourceTracker was used to identify the contribution of the work microbiome to the human microbiome.
Median endotoxin levels ranged from undetectable to 1.0 EU/m3. Significant differences in mouse allergen levels, bacterial copy number, microbial richness, and microbial community structure were identified between animal, dirty, middle, and setup cage wash locations. Endotoxin levels had only a moderate correlation with microbial composition. Location within a facility was a stronger predictor of microbial community composition (R2 = 0.41, p = 0.002) than facility. The contribution of the work microbiome to the pre-shift human microbiome of workers was estimated to be 0.1 ± 0.1% for the oral microbiome; 3.1 ± 1.9% for the nasal microbiome; and 3.0 ± 1.5% for the skin microbiome.
The microbial environment of academic animal care facilities varies significantly by location rather than facility. Endotoxin is not a proxy for assessment of environmental microbial exposures using 16S qPCR or 16S rDNA sequencing. The work microbiome contributes to the composition of the nasal and skin microbiome of workers; the clinical implications of this observation should be further studied.
利用内毒素、16S定量聚合酶链反应(qPCR)和16S扩增子测序来描述学术性小鼠研究设施中工作人员的微生物环境。确定工作微生物群是否对工作人员的人类微生物群有影响。
我们在四个学术性小鼠研究设施的动物房、脏笼清洗区、中间区和笼具设置清洗区进行了区域空气采样。脏笼清洗区的10名工作人员在轮班前和轮班后接受了个人空气采样以及鼻腔、口腔和皮肤样本的重复采集。环境样本进行了内毒素、小鼠过敏原、通过16S qPCR检测细菌拷贝数以及通过16S rDNA测序进行微生物鉴定。轮班前和轮班后的人类样本也进行了16S rDNA测序。使用SourceTracker来确定工作微生物群对人类微生物群的贡献。
内毒素水平中位数范围从检测不到到1.0 EU/m3。在动物房、脏笼清洗区、中间区和笼具设置清洗区之间,小鼠过敏原水平、细菌拷贝数、微生物丰富度和微生物群落结构存在显著差异。内毒素水平与微生物组成仅具有中等相关性。设施内的位置比设施本身更能预测微生物群落组成(R2 = 0.41,p = 0.002)。工作微生物群对工作人员轮班前口腔微生物群的贡献估计为0.1±0.1%;对鼻腔微生物群的贡献为3.1±1.9%;对皮肤微生物群的贡献为3.0±1.5%。
学术性动物护理设施的微生物环境因位置而非设施不同而有显著差异。内毒素不能作为使用16S qPCR或16S rDNA测序评估环境微生物暴露的替代指标。工作微生物群对工作人员鼻腔和皮肤微生物群的组成有影响;这一观察结果的临床意义应进一步研究。