Military veterans account for 8% of homeless individuals living in the United States. To highlight associations between history of homelessness and the gut microbiome, we compared the gut microbiome of veterans who reported having a previous experience of homelessness to those from individuals who reported never having experienced a period of homelessness. Moreover, we examined the impact of the cumulative exposure of prior and current homelessness to understand possible associations between these experiences and the gut microbiome. Microbiome samples underwent genomic sequencing and were analyzed based on alpha diversity, beta diversity, and taxonomic differences. Additionally, demographic information, dietary data, and mental health history were collected. A lifetime history of homelessness was found to be associated with alcohol use disorder, substance use disorder, and healthy eating index compared to those without such a history. In terms of differences in gut microbiota, beta diversity was significantly different between veterans who had experienced homelessness and veterans who had never been homeless ( P = 0.047, weighted UniFrac), while alpha diversity was similar. The microbial community differences were, in part, driven by a lower relative abundance of Akkermansia in veterans who had experienced homelessness (mean; range [in percentages], 1.07; 0-33.9) compared to veterans who had never been homeless (2.02; 0-36.8) ( P = 0.014, ancom-bc2). Additional research is required to facilitate understanding regarding the complex associations between homelessness, the gut microbiome, and mental and physical health conditions, with a focus on increasing understanding regarding the longitudinal impact of housing instability throughout the lifespan.IMPORTANCEAlthough there are known stressors related to homelessness as well as chronic health conditions experienced by those without stable housing, there has been limited work evaluating the associations between microbial community composition and homelessness. We analyzed, for the first time, bacterial gut microbiome associations among those with experiences of homelessness on alpha diversity, beta diversity, and taxonomic differences. Additionally, we characterized the influences of diet, demographic characteristics, military service history, and mental health conditions on the microbiome of veterans with and without any lifetime history of homelessness. Future longitudinal research to evaluate the complex relationships between homelessness, the gut microbiome, and mental health outcomes is recommended. Ultimately, differences in the gut microbiome of individuals experiencing and not experiencing homelessness could assist in identification of treatment targets to improve health outcomes., Competing Interests: Dr. Hoisington reports grants from the VA, DOD, and NIH. Dr. Holliday reports grants from the VA, DOD, American Psychological Association, and NIH. Dr. Forster reports grants from the VA, DOD, NIH, and the state of Colorado. Dr. Postolache reports current support from the VA and NIH and the DC Department of Behavioral Health, Washington, DC. In the past, he was also funded by the American Foundation for Suicide Prevention, NARSAD, and FDA. Dr. Lowry is a member of the faculty of the Integrative Psychiatry Institute, Boulder, CO, USA; the Institute for Brain Potential, Los Banos, CA, USA; and Intelligent Health, Ltd., Reading, UK, and reports grants from the VA, NIH, NSF, and Institute for Cannabis Research. Dr. Brenner reports grants from the VA, DOD, NIH, and the state of Colorado; editorial renumeration from Wolters Kluwer; and royalties from the American Psychological Association, Oxford University Press, and the Rand Corporation. In addition, she consults with sports leagues via her university affiliation.