Additional file 1 of Predicting how varying moisture conditions impact the microbiome of dust collected from the International Space Station
Creators: name: Nastasi, Nicholas
name: Bope, Ashleigh
name: Meyer, Marit E.
name: Horack, John M.
name: Dannemiller, Karen C.
Publisher: name: figshare
Year: 2024
Subjects: subject: Microbiology
subject: FOS: Biological sciences

description: Additional file 1: Table S1: Frozen dust sample dates and location. These frozen samples were not vacuumed, instead they were picked from the location and placed into a sterile bag. They were then frozen at -80C until use in this study. Table S2: Fungal and bacterial concentrations for 2-week incubation samples at 25℃ and each ERH condition tested. Table S3: Summary of Satterthwaite two-sample t-test statistics for fungal and bacterial 2-week incubations. Table S4: qPCR values for fungal and bacterial quantities for frozen dust sample and the original dust collected from the ISS vacuum bags. Table S5: Total fungal growth rates for TOW incubations. Values represent the average of the 4 ISS bags collected. Table S6: qPCR values for all TOW samples. Table S7: Effective growth rate constants (k) for TOW at constant (24 h per day) ERH conditions. Table S8: Effective growth constants (R) for all TOW samples. Table S9: Relative growth constants (R/k) for all TOW samples. Table S10: Most common taxa that was present in all sequenced samples sorted by order, genus, and species for bacteria and fungi. Table S11: Adonis values for fungal bray Curtis PCoA analysis for each time-of-wetness condition. Table 12: Adonis statistics for bacterial time of wetness beta diversity measurements. Table S13: Adonis statistics for fungal and bacterial frozen sample comparisons. Frozen samples were compared to original dust samples (from ISS vacuum bag) as well as 2-week incubations at 50%, 85%, and 100% RH. Table S14: Fungal alpha diversity Kruskal–Wallis statistics for richness and Shannon diversity for 2-week incubations at each RH conditions tested. Significant changes in both richness and Shannon diversity compared to the original dust began to occur at 80% RH (Q &lt; 0.05). Table S15: Richness and Shannon diversity Kruskal–Wallis statistics for fungal time-of-wetness samples. Table S16: Kruskal–Wallis test statistics for alpha diversity metrics for all sequenced bacterial samples. Table S17: Kruskal–Wallis test statistics for fungal and bacterial frozen sample comparisons. Table S18: Differential abundance fungal comparison between non-elevated (Original Dust, 50, 60, 70% RH) and elevated (80, 85, 90, 100% RH) after 2-week constant ERH incubations at 25℃. Table S19: Differential abundance fungal comparison between original dust samples and 24-h TOW saturated (100% RH) conditions. There were 52 fungal species more abundant in the original dust compared to 6 species more abundant at saturated conditions. Table S20: Differential abundance fungal comparison between unmodified (50% RH) 24-h TOW samples and 24-h TOW saturated (100% RH) conditions. There were 29 fungal species more abundant in the unmodified condition compared to 10 species more abundant at saturated conditions. Table S21: Differential abundance fungal comparison between high (85% RH) 24-h TOW samples and 24-h TOW saturated (100% RH) conditions. There was 1 fungal species more abundant in the high condition compared to 8 species more abundant at saturated conditions. Table S22: Differential abundance fungal comparison between high (85% RH) for all TOW samples and all TOW saturated (100% RH) conditions. There were 30 fungal species more abundant in the high condition compared to 4 species more abundant at saturated conditions. Table 23: Example of bacterial differential abundance analysis for non-elevated (original dust and 50% RH) and elevated (80, 85, 90, and 100% RH) conditions for 2-week incubations. No bacterial species were found to be more abundant in either condition. This was true for all time-of-wetness incubation comparisons as well (not shown). Additional file 1: Figure S1: A) Bacterial concentration of original dust and at each ERH condition tested (50, 60, 70, 80, 85, 90, and 100%) after two weeks at 25℃. Quantities for each condition represent a total of 36 qPCR measurements from 4 vacuum bags with triplicate physical samples from each bag and triplicate qPCR measurements per sample. B) Time-of-Wetness models for bacterial growth in ISS dust for elevated (85% ERH) and saturated (100% ERH) conditions. Figure S2: Fungal principal coordinate analyses of time-of-wetness samples separated out by elevated (A) and saturated (B) relative humidity conditions. Figure S3: Fungal principal coordinate analyses of time-of-wetness samples separated out by time points of 6 h (A), 12 h (B), 18 h (C), and 24 h (D). Figure S4: Fungal principal coordinate analyses of 50% ERH time-of-wetness samples separated out by sample days. Figure S5: A) Bacterial PCoA plots for constant ERH 2-week incubations. Only original dust, 50%, 80%, 85%, 90%, and 100% ERH samples were sequenced. B) Bacterial PCoA plot for Time-of-Wetness incubations. Only constant 24-h samples for 50% and 100% ERH conditions were performed for days 5, 10, 14, and 21. Both figures represent weighted unifrac distance matrices. Figure S6: Principal coordinate analyses of frozen dust sample returned from the ISS. Frozen samples were compared to original dust samples (from ISS vacuum bag) as well as 2-week incubations at 50%, 85%, and 100% ERH. Fungi PCoA plots used the Bray–Curtis dissimilarity statistics (C), while bacteria used both weighted (A) and unweighted unifrac (B). Figure S7: Frozen sample alpha diversity plots for (A) bacteria and (B) fungi. Frozen dust samples were compared to original dust, 50% ERH 2-week, 85% ERH 2-week, and 100% ERH 2-week incubations. Figure S8: Mean relative (A) and absolute (B) abundance data for fungal genus in constant equilibrium relative humidity (ERH) incubation samples. Original dust was not incubated and represents what was in the dust in the ISS vacuum bags with no ERH exposure. For each ERH condition, samples were incubated for 2 weeks at 25°C. Each condition (including original dust) represents the mean of 12 total dust samples which includes 3 physical triplicates from the 4 ISS vacuum bags used in this study. Figure S9: Mean relative (A) and absolute (B) abundance data for fungal genus of frozen ISS dust and original dust samples. Original dust was not incubated and represents what was in the dust in the ISS vacuum bags with no ERH exposure. Frozen dust samples were collected onboard the ISS via a tweezer (no vacuum), placed in a triple-sealed plastic bag, and stored at -80°C until use in this study. The abundance data represents of a total of 12 samples for original dust samples (3 for each ISS bag) and 8 frozen dust samples (1 for each location sampled).