Repellency of N,N-diethyl-3-methylbenzamide (DEET) during host-seeking behavior of bed bugs (Hemiptera: Cimicidae) in binary choice olfactometer assays.

The bed bug (Cimex lectularius L.) is one of the most prolific and burdensome indoor pests, and suppression of bed bug populations is a global priority. Understanding bed bug behavior is important to the development of new tactics for their control. Major gaps exist in our understanding of how host cues, insecticide resistance, and exposure modality impact the repellency of formulated products to bed bugs. Here, we validate the use of a binary choice olfactometer for assessing bed bug repellency behaviors using N,N-diethyl-3-methylbenzamide (DEET) in a dose-dependent manner, while considering the role of host-associated stimuli (with vs. without CO2), exposure modality (olfactory vs. olfactory and contact), and resistance status (susceptible vs. resistant) on repellency. We observed that host-seeking insecticide-susceptible bed bugs were repelled only when olfactorily exposed to high concentrations of DEET. However, exposure to DEET by contact repelled insecticide-susceptible bed bugs at 100-fold lower dose of DEET. Further, we demonstrate for the first time that insecticide-resistant bed bugs were significantly more responsive to DEET than susceptible bed bugs. We conclude that the 2-choice olfactometer is an effective tool for assessing the behavioral responses of bed bugs to spatial and contact repellents.

Behavioral interactions of bed bugs with long-lasting pyrethroid-treated bed nets: challenges for vector control.

BACKGROUND: Widespread vector control has been essential in reducing the global incidence and prevalence of malaria, despite now stalled progress. Long-lasting insecticide-treated nets (LLINs) have historically been, and remain, one of the most commonly used vector control tools in the campaign against malaria. LLINs are effective only with proper use, adherence, retention and community adoption, which historically have relied on the successful control of secondary pests, including bed bugs. The emergence of pyrethroid-resistant bed bugs in malaria-endemic communities and failure to control infestations have been suggested to interfere with the effective use of LLINs. Therefore, the behavioral interactions of bed bugs with commonly used bed nets should be better understood. METHODS: To investigate the interactions between bed bugs (Cimex lectularius L.) and LLINs, insecticide-susceptible and pyrethroid-resistant bed bugs were challenged to pass through two commonly used LLINs in two behavioral assays, namely host (blood meal)-seeking and aggregation-seeking assays. The proportions blood-fed and aggregated bed bugs, aggregation time and mortality were quantified and analyzed in different bed bug life stages. RESULTS: Overall, both the insecticide-susceptible bed bugs and highly resistant bed bugs showed a varying ability to pass through LLINs based on treatment status and net design. Deltamethrin-treated nets significantly impeded both feeding and aggregation by the susceptible bed bugs. While none of the tested LLINs significantly impeded feeding (passage of unfed bed bugs through the nets) of the pyrethroid-resistant bed bugs, the untreated bed net, which has small mesh holes, impeded passage of fed bed bugs. Mortality was only seen in the susceptible bed bugs, with significantly higher mortality on deltamethrin-treated nets (63.5 ± 10.7%) than on permethrin-treated nets (2.0 ± 0.9%). CONCLUSIONS: Commonly used new LLINs failed to prevent the passage of susceptible and pyrethroid-resistant bed bugs in host- and aggregation-seeking bioassays. The overall low and variable mortality observed in susceptible bed bugs during both assays highlighted the potential of LLINs to impose strong selection pressure for the evolution of pyrethroid resistance. Already, the failure to control bed bug infestations has been implicated as a contributing factor to the abandonment or misuse of LLINs. For the first time to our knowledge, we have shown the potential of LLINs in selecting for resistant secondary pest populations and so their potential role in stalling malaria control programs should be further investigated. The emergence of pyrethroid-resistant bed bugs in malaria-endemic communities may interfere with the effective use of pyrethroid-impregnated bed nets. We assessed the interactions of two bed bug strains with commonly used bed nets using two behavioral assays, namely host (blood meal)-seeking by unfed bed bugs and aggregation-seeking by freshly fed bed bugs. These assays assessed the passage of bed bugs through various bed nets in response to host cues and aggregation stimuli, respectively. Conditioned paper is a section of file folder paper that has been exposed to bed bugs and has been impregnated with feces and aggregation pheromone; it is attractive to aggregation-seeking fed bed bugs. An unconditioned ramp is a similar section of file folder paper that allows bed bugs to traverse the bed net and gain access to a blood-meal source.