Effects of microwave pretreatment on waste activated sludge (WAS) in mesophilic semicontinuous digesters with acclimatized inoculum at solids retention times (SRTs) of 5, 10, and 20 days are presented. Batch digesters determined optimum microwave temperature, intensity, WAS concentration, and percentage of WAS pretreated for highest WAS solubilization (soluble to total chemical oxygen demand ratio [SCOD:TCOD]) and biogas production. Pretreatment results indicated the potential to damage floc structure and release 4.2-, 4.5-, and 3.6-fold higher soluble proteins, sugars, and SCOD:TCODs compared with controls, with nucleic acid release. Pretreatment increased dewaterability and bioavailability of WAS with 20% higher biogas production compared with controls in batch digestion. In semicontinuous digesters, relative (to control) improvements in removals dramatically increased, as SRT was shortened from 20 to 10 to 5 days, with 23 and 26% higher volatile solids removals for WAS pretreated to 96 degrees C by microwave and conventional heating at a 5-day SRT.