The objective of the Rodent Research-7 mission (RR-7) was to study the impact of the space environment on the gut microbiota of two strains of mice and how any changes in-turn affect the immune system metabolic system and circadian or daily rhythms. To this end ten 11-week-old female C57BL/6J and ten 11-week-old female C3H/HeJ mice were flown to the International Space Station on June 29 2018 on SpaceX-15 and housed in two Rodent Habitats. Samples of food swabs from living surfaces and fecal pellets were collected from each animal before launch and regularly during the mission. The mission also involved extended video collection (48 hr video segments per Habitat) to monitor circadian rhythms and on-orbit mass measurement. After 25 days on-orbit half of the mice of each strain were euthanized on the ISS with Ketamine/Xylazine/Acepromazine and cardiac puncture after which carcasses were segmented in three sections and preserved in RNA later. After 75-76 days the remaining 5 animals from each group were euthanized and processed in the same manner. The 25-day dissected carcasses returned on SpX-15 and the 75-day dissected carcasses returned on SpX-16. In addition to the Flight group three ground control groups were also part of the study: Basal (representing the pre-launch state) Vivarium (standard vivarium housing for the same duration of time as flight) and Ground (same habitat in the International Space Station Environment Simulator ISSES). Twenty mice (10 of each strain) were included in each of these control groups which were euthanized and processed on the same schedule and in the same manner as the flight samples. Dissections for tissues from all experimental groups were completed by the PI groups along with NASA s Biospecimen Sharing Program in February 2019. GeneLab received kidneys from thirty-eight C57BL/6J mice: 9 Basal 5 Ground (25 days) 4 Ground (75 days) 5 Flight (25 days) 5 Flight (75 days) 5 Vivarium (25 days) 5 Vivarium (75 days). GeneLab received kidneys from thirty-nine C3H/HeJ mice: 9 Basal 5 Ground (25 days) 5 Ground (75 days) 5 Flight (25 days) 5 Flight (75 days) 5 Vivarium (25 days) 5 Vivarium (75 days). From these kidneys RNA was extracted libraries generated (stranded ribodepleted) and sequenced (target 60 M clusters at PE 98 bp).

The objective of the Rodent Research-7 mission (RR-7) was to study the impact of the space environment on the gut microbiota of two strains of mice and how any changes in-turn affect the immune system metabolic system and circadian or daily rhythms. To this end ten 11-week-old female C57BL/6J and ten 11-week-old female C3H/HeJ mice were flown to the International Space Station on June 29 2018 on SpaceX-15 and housed in two Rodent Habitats. Samples of food swabs from living surfaces and fecal pellets were collected from each animal before launch and regularly during the mission. The mission also involved extended video collection (48 hr video segments per Habitat) to monitor circadian rhythms and on-orbit mass measurement. After 25 days on-orbit half of the mice of each strain were euthanized on the ISS with Ketamine/Xylazine/Acepromazine and cardiac puncture after which carcasses were segmented in three sections and preserved in RNA later. After 75-76 days the remaining 5 animals from each group were euthanized and processed in the same manner. The 25-day dissected carcasses returned on SpX-15 and the 75-day dissected carcasses returned on SpX-16. In addition to the Flight group three ground control groups were also part of the study: Basal (representing the pre-launch state) Vivarium (standard vivarium housing for the same duration of time as flight) and Ground (same habitat in the International Space Station Environment Simulator ISSES). Twenty mice (10 of each strain) were included in each of these control groups which were euthanized and processed on the same schedule and in the same manner as the flight samples. Dissections for tissues from all experimental groups were completed by the PI groups along with NASA s Biospecimen Sharing Program in February 2019. GeneLab received dorsal skin samples from forty C57BL/6J mice: 10 Basal 5 Ground (25 days) 5 Ground (75 days) 5 Flight (25 days) 5 Flight (75 days) 5 Vivarium (25 days) 5 Vivarium (75 days). GeneLab received dorsal skin samples from forty C3H/HeJ mice: 10 Basal 5 Ground (25 days) 5 Ground (75 days) 5 Flight (25 days) 5 Flight (75 days) 5 Vivarium (25 days) 5 Vivarium (75 days). From these skin samples RNA was extracted libraries generated (stranded ribodepleted) and sequenced (target 60 M clusters at PE 98 bp).

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 Space Algae Experiment Verification Test completed a competitive growth selection on mutagenized microalgae in a series of batch cultures that were passaged for three growth cycles. The experiment was conducted with ground control conditions similar to those actually used in spaceflight. Cultures were limited in growth rate due to gas permeable membranes to provide oxygen and carbon dioxide exchange and a lack of agitation to mix the cells throughout the liquid media. Two strains were grown and three biological replicates were completed for each strain. Specific variables (factors) tested were: 1) The effect of UVC mutagenesis was tested by sampling the algae cultures prior to mutagenesis. 2) The sensitivity of different strains was tested by conducting the experiment with a wild-type and cell wall mutant (cw15) strain. 3) The effect of competitive growth was tested by sampling each biological replicate experiment at the end of each growth cycle. 4) The effect of live culture storage in the dark was tested by sampling each growth cycle twice. At the time of passage a sample of cells was pelleted and frozen. The cultures were then stored in the dark in a Cargo Transport Bag (CTB) to simulate storage on the ISS and return of cultures. This factor tested whether cultures could be stored alive in the dark or if samples needed to be fixed at the time of passage in order to get an accurate representation of the genetic variation in each cycle of growth. Paired-end whole genome sequencing was completed for 38 samples: 2 strains pre mutagenesis and 2 strains X 3 biological replicates X 3 growth cycles X 2 storage conditions.

Rubinsicoccus jplei genome a novel genus isolated from JPL spacecraft environment where Mars 2020 subsystems are assembled.

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 on-going Microbial Observatory Experiments on the International Space Station (ISS) revealed the presence of various microorganisms that may be affected by the distinct environment of the ISS. The low-nutrient environment combined with enhanced irradiation and microgravity may trigger changes in the molecular suit of microorganisms leading to increased virulence and resistance of microbes. Proteomic characterization of two Aspergillus fumigatus strains ISSFT-021 and IF1SW-F4 isolated from HEPA filter debris and cupola surface of the ISS respectively is presented along with a comparison to experimentally established clinical isolates Af293 and CEA10. In-depth analysis highlights variations in the proteome of both ISS-isolated strains when compared to the clinical strains. Proteins up-regulated in ISS isolates were involved in oxidative stress response and carbohydrate and secondary metabolism. This report provides insight into possible molecular adaptation of filamentous fungi to the unique ISS environment. Lastly an attempt was made to elucidate plausible causes of the enhanced virulence of both ISS-isolated A. fumigatus strains.

Genome sequencing and assembly of fungal isolates belonging to Penicilliium and Aspergillus genera isolated from International Space Station

Genome sequencing and assembly of lower Eukaryotic isolates belonging to class Tremellomycetes isolated from International Space Station

Draft genomes of various bacterial phyla isolated from different locations on the International Space Station, as part of the Microbial Tracking 2 mission (ISS-MOP).