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The Internal Active Thermal Control System (IATCS) aboard the International Space Station (ISS) contains an aqueous, alkaline fluid (pH 9.5±0.5) that aids in maintaining a habitable environment for the crew. Because microbes have significant potential to cause disease, adverse effects on astronaut health, and microbe-induced corrosion, the presence of both bacteria and viruses within IATCS fluids is of concern. This study sought to detect and identify viral populations in IATCS samples obtained from the Kennedy Space Center as a first step towards characterizing and understanding potential risks associated with them. Samples were concentrated and viral nucleic acids (NA) extracted providing solutions containing 8.87-22.67 μg NA per mL of heat transfer fluid. After further amplification viral DNA and cDNA were then pooled, fluorescently labeled, and hybridized onto a Combimatrix panvira 12K microarray containing probes for ∼1,000 known human viruses.