In dogs, gut microbiome dysbiosis is associated with several health conditions, including gastrointestinal disease. Probiotic supplementation can support a balanced gut microbiome. This study assessed the impact of a probiotic containing a mixture of Lacticaseibacillus casei, Limosilactobacillus fermentum, Levilactobacillus brevis, and Enterococcus faecium on the gut microbiota of six dogs using short-term colonic simulations. Two groups were included, i.e., blank versus supplementation with the test product, and incubated for 48 h. Probiotic-supplemented reactors had significantly greater fermentative activity compared with the blank, as shown by lower pH levels and higher gas pressure after 6 h, 24 h, and 48 h of incubation (p < 0.05 for all). Saccharolytic fermentation also increased, with a significantly higher level of acetate at 24 h and propionate at 6 h, 24 h, and 48 h with the test product versus blank (p < 0.05 for all). There was no significant effect of the test product on alpha-diversity, but beta-diversity analysis revealed a clear separation in the microbial community composition between the test product and blank. Eight bacterial taxa were enriched with test product supplementation, including the probiotic test strains as well as Megamonas and Bacteroides species. This study, using in vitro short-term colon simulations with six canine donors, provides insights into the probiotic characteristics of the test product.