The dusky-footed woodrat, Neotoma fuscipes Baird is a natural reservoir of the Lyme disease spirochete, Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner, in California. To investigate the potential of host-targeted insecticide to control the tick vectors of B. burgdorferi, permethrin-impregnated or untreated cotton balls were distributed in metal cylinders as potential nesting material adjacent to 95 woodrat houses in chaparral-covered rangeland. Laboratory experiments demonstrated that adult woodrats would enter the cylinders and construct nests from permethrin-treated or untreated cotton. The residual concentration of permethrin did not vary significantly during an 11-mo period and remained > 60% of the registered insecticidal formulation (7.5% [AI] by cotton weight). The abundance of 4 species of ticks (Ixodes neotomae Cooley; the western blacklegged tick I. pacificus Cooley & Kohls; I. woodi Bishopp; and the Pacific Coast tick, Dermacentor occidentalis Marx) infesting woodrats was similar in the treatment and control areas. Although > 90% of the cotton disappeared from the metal cylinders in both areas, examination of 8 active woodrat houses revealed that small amounts of cotton had been incorporated into the nest cups of only 25%. In contrast, the abundance of the flea Orchopeas sexdentatus (Baker) decreased significantly in the treatment area only. Spirochetes were not detected in 168 adult O. sexdentatus fleas that had fed on spirochetemic woodrats, which demonstrates that this flea is an inefficient host of B. burgdorferi. We conclude that the use of permethrin-impregnated cotton as potential nesting material is ineffective for controlling ticks associated with the dusky-footed woodrat in brushlands, but this methodology may be useful for reducing populations of sylvatic fleas.