We investigated the nematicidal and insecticidal activities of the aqueous extract from Bidens pilosa var. radiata on pine-wood nematode (Bursaphelenchus xylophilus) and Japanese pine sawyer (Monochamus alternatus) larvae and adults, the causal agent and vector of pine wilt, respectively. The aqueous extract killed the pine-wood nematode, adults, and larvae of the vector in vitro at all concentrations tested, and the effect decreased significantly with increasing extract dilution. Repellent activity was observed on the nematode and the vector adults as well, although the activity decreased with time in case of the vector. Furthermore, the aqueous extract of B. pilosa effectively promoted approximately 3.0–9.0 mm of hyphal growth in Beauveria spp., when compared with the control treatment. A minor insecticidal effect was also observed on two species of click beetle (Cryptalaus larvatus pini and Paracalais berus), which are natural predators of the Japanese pine-sawyer larvae. Pesticidal and repellent activities of the aqueous extract observed on several organisms related to pine wilt suggest that a multifactorial approach may effectively control this devastating disease.