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Introduction: Non-alcoholic fatty liver disease (NAFLD), the hepatic feature of the metabolic syndrome, is the most prevalent chronic liver diseases in developed countries and urban population of China.Recent studies indicated the modification of gut microbiota plays an important role in the progression from simple steatosis to steatohepatitis.Epidemiological studies have demonstrated consumption of fish oil rich in n-3 polyunsaturated fatty acids such as docosahexanoic acid and eicosapentanoic acid protected against NAFLD progression, but the mechanisms remained unclear.In the present study, we adopted 16s rRNA amplicon sequencing technique to investigate the impacts of fish oil supplement on gut microbiomes in high fat diet (HFD) induced rat NAFLD model.