Microbial contamination during long-term confinements of space exploration presents potential risks for both crew members and spacecraft life support systems. A novel swab kit was used to sample various surfaces from a submerged, closed, analog habitat to characterize the microbial populations. Samples were collected from various locations across the habitat which were constructed from various surface materials (linoleum, dry wall, particle board, glass, and metal), and microbial populations were examined by culture, quantitative PCR (qPCR), microbiome 16S rRNA gene sequencing, and shotgun metagenomics. Propidium monoazide (PMA)-treated samples identified the viable/intact microbial population of the habitat. The cultivable microbial population ranged from below the detection limit to 10⁶ CFU/sample, and their identity was characterized using Sanger sequencing. Both 16S rRNA amplicon and shotgun sequencing were used to characterize the microbial dynamics, community profiles, and functional attributes (metabolism, virulence, and antimicrobial resistance). The 16S rRNA amplicon sequencing revealed abundance of viable (after PMA treatment) Actinobacteria (Brevibacterium, Nesternkonia, Mycobacterium, Pseudonocardia, and Corynebacterium), Firmicutes (Virgibacillus, Staphylococcus, and Oceanobacillus), and Proteobacteria (especially Acinetobacter) on linoleum, dry wall, and particle board (LDP) surfaces, while members of Firmicutes (Leuconostocaceae) and Proteobacteria (Enterobacteriaceae) were high on the glass/metal surfaces. Nonmetric multidimensional scaling determined from both 16S rRNA and metagenomic analyses revealed differential microbial species on LDP surfaces and glass/metal surfaces. The shotgun metagenomic sequencing of samples after PMA treatment showed bacterial predominance of viable Brevibacterium (53.6%), Brachybacterium (7.8%), Pseudonocardia (9.9%), Mycobacterium (3.7%), and Staphylococcus (2.1%), while fungal analyses revealed Aspergillus and Penicillium dominance.

中文翻译：

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有人居住期间封闭栖息地的微生物组和基因组分析。

在太空探索的长期限制中，微生物污染对机组人员和航天器生命支持系统均构成潜在风险。一种新颖的拭子套件用于从淹没的，封闭的，类似的栖息地采样各种表面，以表征微生物种群。从整个栖息地的各个位置收集样品，这些位置由各种表面材料（油毡，干墙，刨花板，玻璃和金属）制成，并通过培养，定量PCR（qPCR），微生物组16S rRNA基因测序检查了微生物种群和shot弹枪宏基因组学。单叠氮化丙锭（PMA）处理的样品确定了栖息地的可行/完整微生物种群。可培养的微生物种群范围从检测限以下到10 ⁶CFU /样品及其身份，使用Sanger测序进行表征。16S rRNA扩增子和shot弹枪测序均用于表征微生物动力学，群落概况和功能属性（新陈代谢，毒力和抗菌素耐药性）。该的16S rRNA扩增子测序揭示了可行的丰度（PMA处理后），放线菌（短杆菌，Nesternkonia，分枝杆菌，假，和棒状杆菌），厚壁菌门（Virgibacillus，金黄色葡萄球菌，和Oceanobacillus）和变形菌（尤其是油毡，干壁和刨花板（LDP）表面上的不动杆菌（Acinetobacter），玻璃/金属表面上的Firmicutes（Leuconostocaceae）和Proteobacteria（Enterobacteriaceae）的成员很高。由16S rRNA和宏基因组学分析确定的非度量多维标度显示LDP表面和玻璃/金属表面上存在差异性微生物。样品的PMA处理后的宏基因组鸟枪测序发现存活的细菌优势杆菌（53.6％），Brachybacterium（7.8％），假（9.9％），结核分枝杆菌（3.7％）和金（2.1％），而真菌分析显示曲霉菌和青霉菌占优势。