Methods for Testing Repellents Against Bed Bugs (Hemiptera: Cimicidae).

Bed bug repellents should not only prevent humans from being bitten but impede an infestation of personal belongings. Only a few test proposals for evaluating the efficacy of repellents against bed bugs have been published so far. In the present study, two test systems were assessed for efficacy testing with five potential bed bug repellents (cinnamon oil, icaridin, N,N-diethyl-3-methylbenzamide (DEET), permethrin, and margosa extract). The first test setup was a harborage choice test system that consisted of a crystallizing dish with a treated and an untreated harborage. Sixty minutes and 24 h after treatment, DEET, icaridin, and cinnamon oil showed the highest repellency with a median proportion of at least 99% repelled bed bugs. The second test system was a barrier test. Bed bugs were attracted by CO2 and heat to cross filter papers treated with the potential repellents. The repellency of substances was significantly lower in comparison to the harborage choice test, except for DEET. The latter showed the highest repellency (97%) against bed bugs 24 h after application compared to controls. Results show that bed bugs are less sensitive to repellents when searching for a bloodmeal than when searching for a shelter.

Detection of target-site and metabolic resistance to pyrethroids in the bed bug Cimex lectularius in Berlin, Germany.

Knockdown-resistance (kdr) against pyrethroids in bed bugs (Cimex lectularis) is associated with the presence of several point mutations in the voltage-sensitive sodium channel α-subunit gene and/or an increased metabolic detoxification by cytochrome P450 monooxygenases (CYPs). In the present study, pyrosequencing assays were developed to quantify the presence of the kdr substitutions (V419L or L925I substitution) in bed bugs in Berlin, Germany. In 14 of 17 bed bug field strains, pyrosequencing revealed the presence of the substitution L925I with allele frequencies between 30% and 100%. One field strain additionally carried the substitution V419L with allele frequencies of 40% in males and 96% in females. In seven of the 17 field strains, mRNA levels of four CYP genes were examined using RT-qPCR. Relative to a susceptible laboratory reference strain, five field strains showed significantly higher mRNA levels of cyp397a1 with 7.1 to 56-fold increases. One of these strains additionally showed a 4.9-fold higher mRNA level of cyp398a1 compared to the reference strain, while cyp4cm1 and cyp6dn1 showed no significant differences. Our findings indicate that multiple resistance mechanisms are present in German C. lectularius populations simultaneously.

A Novel Simulated-Use Test for Determining the Efficacy of Insecticides Against Bed Bugs (Hemiptera: Cimicidae).

The common bed bug Cimex lectularius L. has undergone a worldwide expansion in recent years, due to increased opportunities for dispersal and development of insecticide resistance. For successful control, efficacy testing of products against bed bugs and determination of insecticide resistance under practical conditions are of outstanding importance. A new test system mimicking the practical use situation of residual insecticides was developed and evaluated. Bed bugs were attracted by CO2 and heat to cross surfaces treated with alpha-cypermethrin and bendiocarb. In contrast to the complete efficacy of alpha-cypermethrin (less than 1% surviving bed bugs [with one exception of 5%]), only 45.3, 46, and 29% of insecticide-susceptible bed bugs showed lethal damage 7 d after contact with freshly bendiocarb-treated wallpaper or insecticide aged for 1 or 2 wk. Results show that the efficacy of different insecticides can be assessed with this new test system. Moreover, susceptibility to deltamethrin of five bed bug field strains, collected from infested apartments in Berlin, Germany, was determined in a filter paper contact bioassay. Resistance ratios (RRs) ranged between 4.3 and 20.7. In the novel simulated-use test, efficacy of alpha-cypermethrin was tested against the bed bug strain with the highest RR. In contrast to the insecticide-susceptible laboratory strain, alpha-cypermethrin was not effective against the field strain, where 26-50% of the bed bugs survived and even laid eggs. These results provide evidence for the presence of practically relevant pyrethroid resistance in bed bugs in Germany.