INTERNATIONAL SPACE STATION MICROBIAL OBSERVATORY OF PATHOGENIC VIRUS, BACTERIA, AND FUNGI (ISS-MOP) PROJECT WILL GENERATE THE MICROBIAL CENSUSES ASSOCIATED WITH VARIOUS ISS MODULES USING TRADITIONAL CULTURE-BASED METHODS AND STATE-OF-THE ART MOLECULAR TECHNIQUES. THE OBJECTIVES OF THE ISS-MOP PROJECT ARE TO MEASURE PRESENCE OF VIRAL AND SELECT BACTERIAL AND FUNGAL PATHOGENS AND CORRELATE THEIR PRESENCE ON CREW. THE EFFECTS OF THE SPACEFLIGHT ENVIRONMENT ON VIRAL AND MICROBIAL PATHOGEN DYNAMICS ARE LARGELY UNKNOWN AND REPRESENT BOTH A SIGNIFICANT GAP IN OUR KNOWLEDGE AND AN IMPORTANT OPPORTUNITY TO STUDY THE PRESENCE OF MICROBIAL POPULATIONS AND PREDICT HEALTH RISKS DURING LONG-TERM SPACE EXPLORATION. THE HYPOTHESIS OF THE ISS-MOP PROJECT IS THAT CERTAIN SHORT-LIVING VIRUSES ASSOCIATED WITH INERT ENVIRONMENTAL SPACES IN THE CLOSED HABITAT OF THE ISS CAN HAVE AN ADVERSE INFLUENCE ON CREW HEALTH. TO TEST THIS HYPOTHESIS THE TEAM WILL CATALOGUE VIRUSES ASSOCIATED WITH SURFACES AND AIR OF THE ISS MODULES USING THE ALL-ENCOMPASSING LAWRENCE LIVERMORE MICROBIAL DETECTION ARRAY (LLMDA), WHICH CONTAINS PROBES TO DETECT NOT ONLY VIRUSES BUT ALSO CLINICALLY IMPORTANT HUMAN BACTERIAL AND FUNGAL PATHOGENS. IN ADDITION, SKIN AND SALIVA SAMPLES FROM CREWMEMBERS WILL BE REQUESTED TO TEST THE HYPOTHESIS. THERE ARE THREE AIMS PROPOSED IN THIS ISS-MOP PROJECT. WHILE THE CULTIVATION-BASED ASSAY (AIM 1) PROVIDES A VALID ESTIMATION OF MICROBIAL PRESENCE, IT OFFERS ONLY A LIMITED ASSESSMENT OF THE PHYLOGENETIC/PATHOGENIC DIVERSITY AND PHYSIOLOGICAL BREADTH OF THE MICROBIAL POPULATION PRESENT. THEREFORE, THE TEAM WILL USE THE NOVEL AND COMPREHENSIVE LLMDA MICROARRAY, THAT IS SPECIFICALLY DESIGNED TO ELUCIDATE THE PATHOGENIC VIRAL AND MICROBIAL DIVERSITY PROFILES OF COMPLEX SAMPLES TO AT LEAST THE SPECIES LEVEL (AIM 2). IN ADDITION, WE WILL CHARACTERIZE THE VIRULENCE AND ANTIBIOTIC RESISTANCE MECHANISMS ASSOCIATED WITH THESE PATHOGENS FROM THE ISS BY DEVELOPING A NEW VIRULENCE AND ANTIBIOTIC RESISTANCE GENE ARRAY AND ANALYZE THE SAMPLES ON THIS ARRAY. THE FINAL DELIVERABLE OF THE PROPOSED PROJECT TO NASA WILL BE THE ISS-MOP DATASET, A COMPILATION OF ALL OF THE GENOMIC SEQUENCES AND GENETIC INFORMATION OF VIRUSES AND MICROBES ENCOUNTERED ON AND WITHIN THE ISS HABITAT (AIM 3). THE OBJECTIVES OF THE ISS-MOP PROJECT DIRECTLY RESPOND TO THE NATIONAL RESEARCH COUNCIL COMMITTEE FOR THE DECADAL SURVEY ON BIOLOGICAL AND PHYSICAL SCIENCES IN SPACE RECOMMENDATION P1 WHERE IT STATES, TO ESTABLISH A MICROBIAL OBSERVATORY PROGRAM ON THE ISS TO CONDUCT LONG-TERM, MULTIGENERATIONAL STUDIES OF MICROBIAL POPULATION DYNAMICS. THE ISS-MOP PROJECT WILL ALSO PROVIDE ANSWERS TO NASA SPACE BIOLOGY GUIDING QUESTIONS: HOW DO ALTERATIONS IN GRAVITY AFFECT IN MICROBIAL CELLS (CMM1)?; DOES THE ENHANCED VIRULENCE OR OTHER GENETIC CHANGES OBSERVED WITH MICROBIAL CULTURES FLOWN IN SPACE OCCUR IN OTHER SPECIES (CMM4)?; AND WHAT ARE THE GENETIC CHANGES UNDERLYING PHYSIOLOGICAL RESPONSES TO SPACE FLIGHT DEMONSTRATED IN BIOLOGICAL SYSTEMS (CMM5)?