SpaceX Inspiration4 Oral, Nasal, and Skin Metagenomic and Metatranscriptomic Microbial Swabs

The SpaceX Inspiration4 mission was a 3-day mission with four private astronauts to low Earth orbit that occurred in September 2021. The crew collected biospecimen samples before, during, and after flight. One of these biospecimen collections included oral, nasal, and skin microbial swabs collected from ten locations (oral, nasal cavity, post-auricular, axillary vault, volar forearm, occiput, umbilicus, gluteal crease, glabella, and toe web space). Skin swabs were first dipped in nuclease-free water before swabbing each region for 30 seconds. Oral and nasal swabs did not include this swab-wetting step. Nuclease-free water samples were sequenced as controls. Swabs were collected at three pre-flight timespoints (L-92, L-44, L-3), two in-flight timepoints (flight day 2 (FD2), and flight day 3 (FD3)), and three post-flight timepoints (R+1, R+45, R+82). Swabs were stored in 400uL of Zymo Research DNA/RNA Shield at 4C until nucleic acid extraction. DNA and RNA were extracted from the same swab to generate metagenomic and metatranscriptomic profiles.

SpaceX Inspiration4任务是2021年9月执行的一项为期3天的近地轨道飞行任务，搭载四名私人宇航员。该机组在飞行前、飞行期间及飞行后三个阶段采集了生物标本样本。其中一组生物标本采集包含从10个部位（口腔、鼻腔、耳后、腋窝穹窿、掌侧前臂、枕部、脐部、臀沟、眉间以及趾间隙）采集的口腔、鼻腔及皮肤微生物拭子。具体操作细节如下：皮肤拭子需先浸入无核酸水（nuclease-free water）中，随后在对应区域擦拭30秒；口腔拭子与鼻腔拭子则无需此润湿步骤。无核酸水样本将作为对照样本进行测序。拭子的采集时间点共设8个：3个飞行前时间点（L-92、L-44、L-3）、2个飞行中时间点（飞行第2天（FD2）与飞行第3天（FD3））以及3个飞行后时间点（R+1、R+45、R+82）。所有拭子均保存于400微升Zymo Research出品的DNA/RNA Shield保存液中，置于4℃环境下存储，直至开展核酸提取操作。研究人员从同一拭子中提取DNA与RNA，以生成宏基因组（metagenomic）与宏转录组（metatranscriptomic）谱学特征数据。