Abstract:Background: Data on ovicidal activity of pediculicides are essential to determine the efficacy of commercial products, and to define application schemes. We present an adaptation of historical methods for rearing head lice, and describe their development ex vivo.Methods: Adult head lice were collected and fed on volunteers to obtain fertile eggs of known age. Two methods of feeding were applied: interval feeding (performed every 8-12 hours), and continuous feeding (placing lice in a round Plexiglas receptacle, closed with gauze attached to the skin). Eggs were incubated at 25-28,6°C and 51,8% -69,3% relative humidity. The following outcome measures were used: appearance of eye spot; presence of embryonic structures; presence of embryonic movements; and hatching. A total of 340 eggs were obtained by continuous feeding. Of these, 205 were examined daily. The remaining 135 eggs and those obtained by interval feeding (n=159) were observed once, to confirm hatching status.Results: Eye spots of the embryos started to appear 6 days after oviposition (median = 7 days), and embryonic movements were first seen 9 days after oviposition (median =11 days). Interval feeding of head lice revealed a hatch rate of 10,7% (17/159), as compared to a hatch rate of 80,9% (275/340) achieved with continuous feeding. Hatching started 9 days after oviposition (median = 12 days). Skin irritation (papular rash; moderate to severe itching) appeared after continuous feeding of adult lice on volunteers.Conclusions: Continuous feeding of head lice on a human host is a suitable approach to obtain head lice eggs for ovicidal testing. The method is simple and cheap, offers conditions similar to the natural habitat, and hatch rates are high. However, side effects on volunteers are considerable.