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.

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).

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.

In an on-going Microbial Observatory experimental investigation on the International Space Station (ISS) multiple bacterial isolates of Biosafety Level 2 (BSL-2) were isolated and identified. The antibiotic susceptibility pattern was tested in these BSL-2 isolates for the following antibiotics: cefazolin ciprofloxacin cefoxitin erythromycin gentamycin oxacillin penicillin rifampin tobramycin and many of the BSL-2 isolates showed multiple drug resistance. Among these isolates 21 strains were chosen for whole genome sequencing (WGS) for a possible lead to develop appropriate countermeasures. In addition the genomic data would enable to determine the influence of microgravity on the pathogenicity and virulence in the BSL-2 microorganisms.

In an on-going Microbial Observatory experimental investigation on the International Space Station (ISS) multiple bacterial isolates of Biosafety Level 2 (BSL-2) were isolated and identified. The antibiotic susceptibility pattern was tested in these BSL-2 isolates for the following antibiotics: cefazolin ciprofloxacin cefoxitin erythromycin gentamycin oxacillin penicillin rifampin tobramycin and many of the BSL-2 isolates showed multiple drug resistance. Among these isolates 21 strains were chosen for whole genome sequencing (WGS) for a possible lead to develop appropriate countermeasures. In addition the genomic data would enable to determine the influence of microgravity on the pathogenicity and virulence in the BSL-2 microorganisms.

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.

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 25 xcb x9aC. 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 (102 to 103 CFU/m2). The PMA-qPCR analysis showed considerable variation of viable bacterial population (105 to 109 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.

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

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.

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.