Responses of the northern fowl mite (NFM), Ornithonyssus sylviarum (Canestrini & Fanzago) (Acari: Macronyssidae), and the chicken body louse (CBL), Menacanthus stramineus (Nitzsch) (Phthiraptera: Menoponidae), to variation in temperature, light, and humidity were assessed in bioassays. The location on a continuous thermal gradient at which each ectoparasite arrested was recorded and analyzed. NFM adults arrested at an average temperature of 30.09 ± 0.34°C. Adult CBL and first-instar CBL nymphs arrested at 33.69 ± 0.20°C and 34.99 ± 0.26°C, respectively. Groups of each ectoparasite were placed into clear glass vials (n = 10/vial) with one half shaded, and vials were exposed to three light levels, as follows: high (200 µmolm ^-2s^-1) , low (4 µmolm ^-2s^-1), and nearly no light (0 µmolm ^-2s^-1). The vial cap edges provided an opportunity to assess the interactive effect of light with harborage. NFM avoided light and sought harborage. In low light, the harborage preference overrode the tendency to avoid light. CBL avoided the harborage and showed a minimal preference for light. A four-level humidity gradient was established in two separate experimental arenas for NFM and CBL. Trials were run in ambient light (4 µmolm ^-2s^-1) for the NFM and in nearly no light for the CBL. The NFM gradient used 38 ± 2%, 54 ± 7%, 73 ± 3%, and 90 ± 4% RH, whereas the CBL gradient used 42 ± 5%, 48 ± 7%, 63 ± 4%, and 73 ± 5% RH. NFM showed no humidity response in the walking bioassay, but the CBL settled at the lowest humidity level. Temperature and humidity on different hen body regions were related to the bioassay results and observed on-host ectoparasite distributions.