It is known that high-fat diet and alcohol intake can modulate the gut microbiota and consequently affect physiological processes such as fat storage and conditional behavior. However, the effects of the interaction between high-fat diet, its withdrawal and ethanol intake in gut microbiota remain unclear. To address this question, we used an animal model in which C57BL/6 mice were fed on standard (AIN93G) or high-sugar and -butter (HSB) diet for 8 weeks. Then, a protocol of free choice between water and a 10% alcohol solution was introduced, and the HSB diet was replaced with AIN93G in two experimental groups. This model allowed us to distinguish the individual effects of HSB diet and ethanol, and the effects of its interaction on the microbiome. The interaction of those factors was the main driver in the structure changes of the fecal microbial community. HSB diet and ethanol consumption directly affected the abundance of Firmicutes and Actinobacteria phylum, and Clostridiaceae and Coriobacteriaceae family. On the other hand, we also showed that abundance of Bacteroidales_S24-7 family and the Firmicutes/Bacteroidetes ratio were affected only by HSB diet consumption and that ethanol consumption was uniquely responsible for the bacterial translocation to the liver, indicating a breaking of the gut barrier. Finally, we also pointed out that the withdrawal of the HSB diet affects the preference for alcohol and shows a structural resilience in the fecal microbiome. These results highlight the importance of the gut microbiome modulation and its possible role on the phenotype developed by animals.

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