The pretreatment of seaweed by washing in freshwater is often used in seaweed biofuel research studies. However, the effect of washing seaweed prior to anaerobic digestion (AD) does not appear to have been greatly studied. This study examines washing Sargassum muticum with freshwater and its effect on ultimate and proximate analyses, salt content, methane production from anaerobic digestion, and leachate loss from ensiling. Washing with freshwater significantly (p < 0.01) increased moisture content (unwashed 85.6%, washed 89.1%) but significantly (p < 0.05) reduced ash (unwashed 32.7% dry weight dw, washed 30.6% dw) and salt content (unwashed ash containing 51.5%, washed 42.5%). The dry biomass higher heating value was significantly (p < 0.05) increased by washing due to the lower ash content (11.5 to 12.6 kJ g−1 dw). There was no significant change in the protein or lipid content, although washing increased the nitrogen content (3.85–4.77% dw). Washing significantly (p < 0.05) increased leachate losses during ensiling, with total leachate losses increasing after washing (12.7–25.2%). The methane yield from anaerobic digestion (28 days) was not statistically significantly different (p > 0.05) between unwashed (0.225 L CH4 g−1 VS) and washed samples (0.177 L CH4 g-1 VS). However, washing delayed biomethane production.