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A large‐arena bioassay is used to examine sex differences in spatiotemporal patterns of bed bug  Cimex lectularius  L. behavioural responses to either a human host or CO 2  gas. After release in the centre of the arena, 90% of newly‐fed bed bugs move to hiding places in the corners within 24 h. They require 3 days to settle down completely in the arena, with generally low activity levels and the absence of responses to human stimuli for 5 days. After 8–9 days, persistent responses can be recorded. Sex differences are observed, in which females are more active during establishment, respond faster after feeding, expose themselves more than males during the daytime, and respond more strongly to the host signal. The number of bed bugs that rest in harbourages is found to vary significantly according to light setting and sex. Both sexes stay inside harbourages more in daylight compared with night, and males hide more than females during the daytime but not during the night. The spatial distribution of the bed bugs is also found to change with the presence of CO 2 , and peak aggregation around the odour source is observed after 24 min. Both male and female bed bugs move from hiding places or the border of the arena toward the centre where CO 2  is released. Peak responses are always highest during the night. Bed bug behaviour and behaviour‐regulating features are discussed in the context of control methods.

Questing activity in bed bug populations: male and female responses to host signals