Chronic alcohol consumption is associated with perturbation of hepatic metabolism of sulphur-containing amino acid. The goal of present study was to evaluate the influence of dietary supplementation of methionine or folate to chronically ethanol-fed mts on the metabolism of sulfur-containing amino acids and one-carbon metabolism. Sprague-Dawley male mts were fed Lieber-Decarli liquid diet with 0% ethanol (control), 36% ethanol (E), 36% ethanol combined with methionine supplement (EM) or folate supplement (EF) for 8 weeks. Hepatic S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH), plasma folate and homocysteine (Hcy), urinary excretion of folate and formiminoglutamate were investigated after feeding experimental diets. Growth was retarded by 36% ethanol consupmtion (E, EM and EF) (p<0.01). Liver total fat (p<0.05) and plasma ALT (P<0.01) were increased by methionine supplementation (EM), implicating fatty liver and liver injury. Liver folate was increased slightly by folate supplementation (EF) (p=0.077). Urinary folate loss was increased 2.3 fold by ethanol consumption (E) and 17.2 fold by folate supplementation (EF), while decreased by methionine supplementation (EM) (p<0.000l). Plasma Hcy was increased 1.9 fold by methionine supplementation (EM) in ethanol-fed mts (p<0.05), which was related with decreased methionine synthase activity (p<0.05). Hepatic SAM/SAH ratio was depressed by methionine supplementation in ethanol-fed mts (EM) (p<0.05). Urinary formininoglutamate (Figlu) excretion after histidine loading was increased by ethanol ingestion and reduced by methionine supplementation (p<0.00l). Based on these data, methionine supplementation appears to accelerate histidine oxidation. In conclusion, dietary supplementation of methionine to ethanol-fed mts exacerbates alcoholic liver injury possibly by complicating sulphur-containing amino acid metabolism, as while it may have beneficial effects on folate and histidine metabolism.