Ultrasound pretreatment was proved to be effective in accelerating waste activated sludge (WAS) disintegration and promoting anaerobic fermentative short-chain fatty acids (SCFA) production. However, due to the high activities of SCFA consumers, SCFA yield is limited. Herein we reported an efficient strategy, i.e., using free ammonia (FA) to aid ultrasound pretreatment, to enhance SCFA accumulation. Experimental results showed that the greatest SCFA yield of 316.7 +/- 14.9 mg COD/g VSS was obtained at 2 w/mL ultrasound pretreatment for 15 min followed by initial 60 mg/L FA pretreatment for 2 d, which was 2.2 times (1.7 times) of that from sole ultrasound (FA) pretreatment. The mechanism study revealed that this combination method accelerated sludge solubilization, enhanced the release of biodegradable organics, and caused severe inhibitions to SCFA consumption. Moreover, this method has inhibitory effects on specific activities of hydrolytic microorganisms, SCFA-producing bacteria and methanogens, but the inhibitions to methanogens were severe.