Genomic and proteomic characterization of the Aspergillus niger isolate JSC-093350089 collected from U.S. segment surfaces of the International Space Station (ISS) is reported along with a comparison to the experimentally established strain ATCC 1015. Whole-genome sequencing of JSC-093350089 revealed enhanced genetic variance when compared to publicly available sequences of A. niger strains. Analysis of the isolate xe2 x80 x9a xc3 x84 xc3 xb4s proteome revealed significant differences in the molecular phenotype of JSC-093350089 including increased abundance of proteins involved in the A. niger starvation response oxidative stress resistance cell wall integrity and modulation and nutrient acquisition. Together these data reveal the existence of a distinct strain of A. niger onboard the ISS and provide insight into the molecular phenotype that is selected for by melanized fungal species inhabiting spacecraft environments.

The first global genomic proteomic and secondary metabolomic characterization of the filamentous fungus Aspergillus nidulans following growth onboard the International Space Station (ISS) is reported. The investigation included the A. nidulans wild-type and three mutant strains two of which were genetically engineered to enhance secondary metabolite production. Whole genome sequencing revealed that ISS conditions altered the A. nidulans genome in specific regions. In strain CW12001 which features overexpression of the secondary metabolite global regulator laeA ISS conditions induced the loss of the laeA stop codon. Differential expression of proteins involved in stress response carbohydrate metabolic processes and secondary metabolite biosynthesis was also observed. ISS conditions significantly decreased prenyl xanthone production in the wild-type strain and increased asperthecin production in LO1362 and CW12001 which are deficient in a major DNA repair mechanism. These data provide valuable insights into the adaptation mechanism of A. nidulans to spacecraft environments.

The first global genomic, proteomic, and secondary metabolomic characterization of the filamentous fungus Aspergillus nidulans following growth onboard the International Space Station (ISS) is reported. The investigation included the A. nidulans wild-type and three mutant strains, two of which were genetically engineered to enhance secondary metabolite production. Whole genome sequencing revealed that ISS conditions altered the A. nidulans genome in specific regions. In strain CW12001, which features overexpression of the secondary metabolite global regulator laeA, ISS conditions induced the loss of the laeA stop codon. Differential expression of proteins involved in stress response, carbohydrate metabolic processes, and secondary metabolite biosynthesis was also observed. ISS conditions significantly decreased prenyl xanthone production in the wild-type strain and increased asperthecin production in LO1362 and CW12001, which are deficient in a major DNA repair mechanism. These data provide valuable insights into the adaptation mechanism of A. nidulans to spacecraft environments.

Genomic and proteomic characterization of the Aspergillus niger isolate, JSC-093350089, collected from U.S. segment surfaces of the International Space Station (ISS) is reported, along with a comparison to the experimentally established strain ATCC 1015. Whole-genome sequencing of JSC-093350089 revealed enhanced genetic variance when compared to publicly available sequences of A. niger strains. Analysis of the isolate’s proteome revealed significant differences in the molecular phenotype of JSC-093350089, including increased abundance of proteins involved in the A. niger starvation response, oxidative stress resistance, cell wall integrity and modulation, and nutrient acquisition. Together, these data reveal the existence of a distinct strain of A. niger onboard the ISS and provide insight into the molecular phenotype that is selected for by melanized fungal species inhabiting spacecraft environments.

Aspergillus fumigatus is a saprophytic filamentous fungus that is ubiquitous outdoors (soil decaying vegetation) and indoors (hospitals simulated closed habitats etc.). A. fumigatus can adapt to various environmental conditions and form airborne conidia that are the inoculum for a variety of diseases (e.g. non- and invasive pulmonary infections allergic bronchopulmonary aspergillosis etc.) in immunocompromised hosts. In an on-going Microbial Observatory Experiments on the International Space Station (ISS) molecular phylogeny of several fungal isolates were characterized. Two strains ISSF 21 and IF1SW-F4 were isolated from the HEPA filter and the surface of the Cupola of the ISS respectively. Using primers targeting the internal transcribed spacers ITS1 and 2 both isolates were identified as A. fumigatus. The whole genome sequence analysis of ISSF 21 revealed increased number of single nucleotide polymorphisms (SNPs) when compared to the reference A. fumigatus 293. Knowing that A. fumigatus is an opportunistic pathogen and microgravity highly influences the antibiotic susceptibility and pathogenicity of microorganisms we examined pathogenicity of both ISS isolates using the zebrafish larval model. The space station isolates (ISSF-021 and IF1SW-F4) were more virulent than two clinical strains (Af293 and CEA10) whose pathogenicity was highly characterized. Here the whole genome sequences of ISSF-021 strain are being deposited.

The International Space Station (ISS) is a unique habitat for humans and microorganisms. Here, we report the results of the ISS experiment EXTREMOPHILES, including the analysis of microbial communities from several areas aboard at three time points. We assess microbial diversity, distribution, functional capacity and resistance profile using a combination of cultivation-independent analyses (amplicon and shot-gun sequencing) and cultivation-dependent analyses (physiological and genetic characterization of microbial isolates, antibiotic resistance tests, co-incubation experiments). We show that the ISS microbial communities are highly similar to those present in ground-based confined indoor environments and are subject to fluctuations, although a core microbiome persists over time and locations. The genomic and physiological features selected by ISS conditions do not appear to be directly relevant to human health, although adaptations towards biofilm formation and surface interactions were observed. Our results do not raise direct reason for concern with respect to crew health, but indicate a potential threat towards material integrity in moist areas.

Presented here is the environmental microbiome study of the International Space Station surfaces. The environmental samples were collected with the polyester wipes from eight different locations in the ISS during two consecutive sampling sessions (three months apart). The specific objective was to unveil the pool of genes for each location during two separate sessions to learn of functional and metabolic diversity of microorganisms in the ISS. The International Space Station (ISS) as a closed built environment has its own environmental microbiome which is shaped by microgravity, radiation, and limited human presence. The microbial diversity associated with ISS environmental surfaces was investigated during this study. Polyester wipes and contact slides were used for sampling of eight various surface locations on the ISS at different time periods. The samples were retrieved and analyzed immediately upon the return to the Earth (via Soyuz TMA-14M or Dragon capsule from SpaceX). After surface sample collection, contact slides containing nutrient media for the growth of bacteria and fungi were incubated at 25C. The polyester wipes were processed to measure microbial burden (R2A, Blood Agar, and Potato Dextrose Agar) and recover cultivable bacteria as well as fungi. Subsequently, viable microbial burden was assessed using Adenosine Triphosphate (ATP) assay, and quantitative polymerase chain reaction (PCR) methods after propidium monoazide (PMA) treatment. The 16S-tag and metagenome analyses were used to elucidate viable microbial diversity. The cultivable bacterial population yield from the polyester wipes was very high (5 to 7-logs) when compared with the contact slides (10^2 to 10^3 CFU/m2). The PMA-qPCR analysis showed considerable variation of viable bacterial population (10^5 to 10^9 16S rDNA gene copies/m2) among locations sampled. Unlike contact slides, polyester wipes cover much larger sample surface (~1 m2) and produce much more reliable results of the microbial diversity of the ISS covering both cultivable and non-cultivable species. The cultivable, total, and viable microbial diversity was determined utilizing state-of-the art molecular techniques. The implementation of the PMA assay before DNA extraction allowed distinguishing viable microorganisms, which is crucial for determining their role to the crew health, the ISS maintenance and the general knowledge of the closed environmentally controlled built systems.

The safety of the International Space Station (ISS) crewmembers and maintenance of ISS hardware are the primary rationale for monitoring microorganisms in this closed habitat. The composition of the microbial community of this built environment is unique due to microgravity space radiation and elevated carbon dioxide levels. As built environments are known to have their own microbiomes next-generation sequencing methods have to be utilized to explore the ISS microbial profile and use this data for further development of safety and maintenance practices. ISS vacuum cleaner bag components (surface) and high-efficiency particulate arrestance (HEPA) filter element (air) samples were analyzed by traditional cultivation adenosine triphosphate (ATP) and propidium monoazide-quantitative polymerase chain reaction (PMA-qPCR) assays to estimate viable microbial populations. In addition vacuum cleaner bag components of two cleanrooms at Jet Propulsion Laboratory (JPL Pasadena CA) were examined concurrently. The 16S rRNA gene sequencing based on Illumina platform was used to elucidate the ISS microbial diversity and explore differences between the microbiomes of the ISS and Earth-based cleanrooms. The statistical analyses of these microbiomes show that Actinobacteria Firmicutes and Proteobacteria dominate in the air and surface of the ISS and the cleanroom samples but vary in abundance. While members of Actinobacteria were predominant in the ISS Proteobacteria the least abundant phylum in the ISS dominated the Earth-based cleanrooms. The viable bacterial population (PMA-treated samples) decreased significantly but the treatment did not appear to have an effect on the bacterial composition (diversity) associated with a sampling site. Viable fungal sequences were not retrieved from the ISS HEPA sample where as highest viable fungal diversity was observed in the Earth-based cleanroom (JPL class 100K) debris. The results of this study provided strong evidence of substantial contribution of human skin-associated microorganisms such as Corynebacterium/Propionibacterium (Actinobacteria),not Staphylococcus (Firmicutes) species as the dominant species in the ISS in terms of viable and total bacterial community structure.

The safety of the International Space Station (ISS) crewmembers and maintenance of ISS hardware are the primary rationale for monitoring microorganisms in this closed habitat. The composition of the microbial community of this built environment is unique due to microgravity space radiation and elevated carbon dioxide levels. As built environments are known to have their own microbiomes next-generation sequencing methods have to be utilized to explore the ISS microbial profile and use this data for further development of safety and maintenance practices. ISS vacuum cleaner bag components (surface) and high-efficiency particulate arrestance (HEPA) filter element (air) samples were analyzed by traditional cultivation adenosine triphosphate (ATP) and propidium monoazide-quantitative polymerase chain reaction (PMA-qPCR) assays to estimate viable microbial populations. In addition vacuum cleaner bag components of two cleanrooms at Jet Propulsion Laboratory (JPL Pasadena CA) were examined concurrently. The 16S rRNA gene sequencing based on Illumina platform was used to elucidate the ISS microbial diversity and explore differences between the microbiomes of the ISS and Earth-based cleanrooms. The statistical analyses of these microbiomes show that Actinobacteria Firmicutes and Proteobacteria dominate in the air and surface of the ISS and the cleanroom samples but vary in abundance. While members of Actinobacteria were predominant in the ISS Proteobacteria the least abundant phylum in the ISS dominated the Earth-based cleanrooms. The viable bacterial population (PMA-treated samples) decreased significantly but the treatment did not appear to have an effect on the bacterial composition (diversity) associated with a sampling site. Viable fungal sequences were not retrieved from the ISS HEPA sample where as highest viable fungal diversity was observed in the Earth-based cleanroom (JPL class 100K) debris. The results of this study provided strong evidence of substantial contribution of human skin-associated microorganisms such as Corynebacterium/Propionibacterium (Actinobacteria),not Staphylococcus (Firmicutes) species as the dominant species in the ISS in terms of viable and total bacterial community structure.

This study investigated the interplay between the microbial community of the International Space Station and its crew. Environmental samples were collected from 8 habitable locations around the ISS. The microbial composition was measured using shotgun metagenomic sequencing and procesed using the Livermore Metagenomics Analysis Toolkit.