Influences of Exposure Time and Mortality Assessment Interval on Bioassay Results of Insecticide-Resistant Tropical Bed Bugs (Hemiptera: Cimicidae)

SIMPLE SUMMARY: Bed bugs cause health and economic impacts due to their hematophagous behavior. The tropical bed bug, Cimex hemipterus (F.) is predominant in tropical and subtropical regions and modern strains demonstrate high levels of insecticide resistance making them a major challenge to control. In this present study, we investigated the effect of exposure time and mortality assessment interval on bioassay results. Bed bugs were treated separately with six insecticide products at label rates using the surface contact method, with multiple exposure times (5 min, 10 min, 30 min, 1 h, 4 h, and continuous exposure for 96 h or 14 days for Phantom). Post-treatment mortalities of bed bugs were assessed daily for up to 4 days (Tandem, Temprid SC, Pesguard FG161, Sumithrin, and Sumithion) and 14 days for Phantom. Results showed that longer exposure times led to rapid knockdown and better killing effect in bed bugs. Exposure times and mortality assessment intervals should be carefully considered in resistance and efficacy studies with insecticide products. ABSTRACT: We evaluated the influences of insecticide exposure time and mortality assessment intervals on the bioassay results of three insecticide-resistant tropical bed bug Cimex hemipterus (F.) populations (Madam Mo, Tanjong Tokong, and Green Lane). This was achieved using the surface contact method and tested with six commercial insecticide products: Tandem, Temprid SC, Pesguard FG161, Sumithrin, Sumithion, and Phantom applied at label rate on glass petri dishes. Six exposure times (5 min, 10 min, 30 min, 1 h, 4 h, and continuous exposure for 4 or 14 days for Phantom) were tested. A susceptible common bed bug Cimex lectularius L. strain (Monheim) was used as the reference strain as no susceptible C. hemipterus strain exists. In treatment with Temprid SC, the Tanjong Tokong strain showed significantly higher KT(50) values at 5, 10, and 30 min exposures than 1 h, 4 h, and continuous exposures. When all resistant populations were evaluated at continuous exposure to Sumithion, they demonstrated moderate resistance levels (RR(50) ranged from 6.0 to 7.9), while KT(50) and KT(95) of other shorter exposure times failed to be generated due to low knockdown rate. Higher mortalities were observed in Tanjong Tokong and Green Lane strains when tested at longer exposure times with Temprid SC, Pesguard FG161, Sumithrin, Sumithion, and Phantom. Better killing effect was observed in the treatment with Temprid SC (Tanjong Tokong and Green Lane strains), Pesguard FG161 (Tanjong Tokong and Green Lane strains), Sumithrin (all C. hemipterus strains), Sumithion (all C. hemipterus strains), and Phantom (all strains tested) at longer mortality assessment intervals. We demonstrated that insecticide exposure time and mortality assessment interval could potentially affect outcomes of product performance evaluations, resulting in underestimation or overestimation of insecticide resistance levels in field populations.