The objective was to evaluate gut microbiome in relation to recent aspartame and Acesulfame‐K artificial sweetener consumption. Thirty‐one adults completed a four‐day food record and provided a fecal sample on the fifth day. Fecal samples were analyzed for bacterial DNA using Multitag Pyrosequencing. Median values for bacterial abundance across non‐consumers and consumers were compared. Overall bacterial abundance profiles across non‐consumers and consumers were compared using UniFrac analysis. Seven participants (23%) consumed aspartame. Seven participants (23%) consumed Acesulfame‐K. These were not the same seven individuals, and three individuals consumed both aspartame and Acesulfame‐K. No participants consumed saccharin. There were no differences in median bacterial abundance for any bacteria (class or order) across consumers and non‐consumers of either aspartame or Acesulfame‐K. The median Bacteriodetes:Firmcutes ratio also did not differ across aspartame non‐consumers (0.96, range: 0.15‐2.97) and consumers (1.08, range: 0.69‐1.87), (p‐value=0.60). Relative abundance of bacteria by class is illustrated for Acesulfame‐K non‐consumers vs. consumers. The median Bacteriodetes:Firmcutes ratio was non‐significantly higher in Acesulfame‐K non‐consumers (1.11, range: 0.14‐2.97) and consumers (0.79, range: 0.38‐2.97), (p‐value=0.74). These results suggest that recent intake of aspartame or Acesulfame‐K is not associated with overall gut microbiome profile in adults. Further studies with more individuals are warranted to evaluate lower abundance microbial taxa or interactions other factors. Support: Internal Funding