<p>The termiticidal activity of bio-oil derived from the wet torrefaction of tobacco (<i>Nicotiana tabacum</i> L) stem was investigated as a potential biopesticide against the drywood termite <i>Cryptotermes cynocephalus</i> (Blattodea: Kalotermitidae). Forced feeding assays were conducted over 21 days using filter paper treated with varying concentrations of bio-oil produced from Virginia and Native tobacco varieties. Efficacy was assessed by comparing termite mortality and filter paper consumption rates in both no-choice and choice feeding tests. The bio-oil demonstrated high toxicity in no-choice tests, with a 2.5% concentration of the Virginia and Native varieties resulting in 78–100 and 83–100% mortality, respectively. Active termite feeding and significant paper consumption was observed in all treatments. In choice feeding tests, lower mortality rates (36–60%) were observed and the continued consumption of treated paper, confirms the non-repellent property of the tobacco bio-oil against <i>C. cynocephalus</i>. The combination of sustained feeding and delayed mortality suggests that the bio-oil acts primarily as a stomach toxicant rather than a fast-acting contact poison. The efficacy is chemically linked to the Native oil’s richness in nicotine and other nitrogen-containing heterocycles, and the Virginia oil’s abundance of oxygenated organic molecules like butanoic acid. Although preliminary, these results require confirmation using solid-wood feeding bioassays under laboratory conditions and subsequent field trials; nevertheless, the findings indicate that tobacco-derived bio-oil has potential as a sustainable biopesticide for termite control.</p>