Water is a critical commodity for spacecraft crews, requiring extreme conservation and reclamation strategies. In addition to suppression of the immune system in spaceflight, enhancement of bacterial growth and antimicrobial resistance in weightlessness raise serious concerns regarding microbial contamination of water systems. Rapid methods are needed for monitoring water, both pre-flight and on orbit. We are developing techniques to enumerate specific, metabolically active bacteria that may threaten crew health or lead to water system deterioration. Our methods are directed at the detection of individual bacteria, rather than populations of bacteria, and we aim to determine the identity of the organism as well as its physiological state con-currently. Our objectives are to determine, in a single test, the total number of bacteria present in a water sample, if a specific strain of bacteria is present within the total population and if these bacteria are viable or dead. We report on the combination of methods into highly sensitive, rapid technologies for detecting specific target bacteria and assessing their viability in spacecraft water.