As one of the near-to-mid-term alternatives to fossil fuels, cellulosic biofuels can cut greenhouse gas emissions while continuing to meet liquid transportation fuel needs. By processing cellulosic biomass into pellets, density and handling efficiency of cellulosic feedstocks will be improved, resulting in a reduction in transportation and handling costs in biofuel manufacturing. Temperature of biomass during the pelleting process can affect the quality of the pellet. But effects of pelleting variables on biomass temperature during ultrasonic vibration-assisted (UV-A) pelleting are still unknown. This paper reports an experimental investigation on temperature of biomass in UV-A pelleting. It studies the effects of moisture content of the biomass and pelleting variables (ultrasonic power, tool travel distance, and feedrate). The results will be helpful in understanding the effects of ultrasonic vibration on biomass temperature, compaction mechanism, and biofuel conversion.