Search

The cryptic behavior that characterizes bed bugs (Cimex lectularius L.) makes them one of the most difficult indoor pests to control. Thus, the use of products with long residual efficacy is a key component of successful bed bug management. Aprehend is a biopesticide for bed bug control, whose active ingredient is the entomopathogenic fungus, Beauveria bassiana. This product shows promise for extended bed bug control against pyrethroid-resistant bed bugs, causing mortality days after contact with the treated surface. However, it is unclear how surface type and distance traveled by bed bugs over a treated surface impact efficacy. Therefore, we evaluated the efficacy of different Aprehend band widths applied to different substrates commonly encountered by bed bugs, looking specifically at average time to mortality and overall mortality. Our findings indicate that exposure to fresh applications of Aprehend resulted in high mortality when bed bugs traveled very short distances (1 cm) over some Aprehend-treated surfaces (vinyl tile and cotton jersey fabric), while exposure to other treated surfaces (unfinished pinewood and painted drywall) resulted in comparatively lower mortality even over longer distances (5 cm). Furthermore, we found that bed bugs crawling on cotton jersey fabric picked up a significantly higher number of spores compared to unfinished pinewood. These results indicate that applications of Aprehend as 5 cm (2 inches) bands according to label recommendations can be effective, although surface type is an important determinator of efficacy. This information will help guide pest management professionals in their use of B. bassiana for bed bug control., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

The common bed bug, Cimex lectularius, is one of the main human parasites. The world-wide resurgence of this pest is mainly due to globalization, and the spread of insecticide resistance. A few studies have compared the transcriptomes of susceptible and resistant strains; however, these studies usually relied on strains originating from distant locations, possibly explaining their extended candidate gene lists. Here, we compared the transcriptomes of 2 strains originating from the same location and showing low overall genetic differentiation (FST=0.018) but varying in their susceptibility to pyrethroids, before and after insecticide exposure. In sharp contrast with previous studies, only 24 genes showing constitutive differential expression between the strains were identified. Interestingly, most of the genes with increased expression in the resistant strain encoded cuticular proteins. However, those changes were not associated with significant difference in cuticular thickness, suggesting that they might be involved in qualitative changes in the cuticle. In contrast, insecticide exposure induced the expression of a multitude of genes, mostly involved in detoxification. Finally, our set of transcriptome candidate loci showed little overlap with a set of loci strongly genetically differentiated in a previous study using the same strains. Several hypothesis explaining this discrepancy are discussed., Competing Interests: Conflict of interest The authors declare no competing interests., (© The Author(s) 2024. Published by Oxford University Press on behalf of Society for Molecular Biology and Evolution.)

Living in groups can provide essential experience that improves sexual performance and reproductive success. While the effects of social experience have drawn considerable scientific interest, commonly used behavioral assays often do not capture the dynamic nature of interactions within a social group. Here, we conducted 3 experiments using a social network framework to test whether social experience during early adulthood improves the sexual competence of bed bugs ( Cimex lectularius ) when placed in a complex and competitive group environment. In each experiment, we observed replicate groups of bed bugs comprising previously socialized and previously isolated individuals of the same sex, along with an equal number of standardized individuals of the opposite sex. Regardless of whether we controlled for their insemination history, previously isolated males mounted and inseminated females at significantly higher rates than previously socialized males. However, we found no evidence of social experience influencing our other measures of sexual competence: proportion of mounts directed at females, ability to overcome female resistance, and strength of opposite-sex social associations. We similarly did not detect effects of social experience on our female sexual competence metrics: propensity to avoid mounts, rate of successfully avoiding mounts, opposite-sex social association strength, and rate of receiving inseminations. Our findings indicate that early social experience does not improve sexual competence in male and female bed bugs., (© The Author(s) 2024. Published by Oxford University Press on behalf of the International Society for Behavioral Ecology.)

Modern bed bugs are resistant to multiple insecticide classes, particularly the pyrethroids. The efficacy of pyrethroid-impregnated mattress liners marketed for bed bug management has been variable. This study evaluated the efficacy of a permethrin-impregnated mattress liner, ActiveGuard, against 24 bed bug strains, consisting of both Cimex hemipterus (F.) and Cimex lectularius L. A 'mat assay', employing an allethrin-impregnated mat, was used to establish the pyrethroid resistance profile of all strains. Three experiments were conducted to evaluate the effect of ActiveGuard exposure on bed bug knockdown: 1) exposing the bed bugs continuously on the liner for up to 24 d, 2) holding the bed bugs on the liner for either 4 or 6 h, and 3) placing a noninsecticide treated fabric above the liner with the bed bugs held continuously on top. Our results indicated that all modern strains (collected within the last 15 years during the current resurgence) were pyrethroid-resistant, although the magnitude of resistance was highly variable between strains. In the continuous exposure study, an incomplete knockdown was recorded for most modern bed bug strains, with some having no knockdown even up to 7 d of constant exposure. In the 4 or 6 h exposure study, the level of knockdown was reduced even further, and very few bed bugs were knocked down in the double fabric study. The results of this study indicate that pyrethroid-impregnated mattress liners are not likely to be effective in the management of most modern bed bug infestations involving either C. hemipterus or C. lectularius., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Bed bugs are an important group of medical and urban insect pests. They are obligate blood-feeders. Their bites may cause skin irritation and allergic reactions and, under some circumstances, may lead to mental and other health issues. Despite numerous discoveries on the biology of these obnoxious pests and progress in control strategies over the last two decades, bed bugs continue to preferentially plague those from low socioeconomic communities because the poor generally could not afford effective control options. As a result, such infestations in poorer communities serve as a reservoir for wider society. This Special Collection of the Perspective on Biology and Management of Bed Bugs presents nine original research papers on bed bug detection, insecticide performance and resistance, nonchemical treatment, fungal biopesticides, and pest management procurement and contracts. We hope that these investigative findings will spur research on safer, more affordable, and effective control options in the future., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The presence of fecal spots has often been used to verify the existence of a bed bug (Cimex lectularius) infestation. However, no research has been conducted to determine how much fecal material that a bed bug population produces over time. In this study, the number of fecal spots that each nymphal life stage was capable of producing after a bloodmeal was quantified. Adult fecal spots were also quantified to determine if there was consistent production between feedings. During this study, it was discovered that bed bugs produced visible fecal spots and clear spots that were only visible under ultraviolet light. Therefore, three types of fecal spots were quantified: dark feces, light feces, and clear spots. Clear spots were produced in greater amounts (38.5-55.5%) than either dark spots (27.3-40.7%) or light spots (17.3-21.9%). For example, 5th instar bed bugs were thought to produce an average of 21 spots (dark and light) after a single bloodmeal. However, using the ultraviolet light, it was found that the 5th instars actually produced an average of 44.7 spots. Using the total fecal spot data collected during this study, researchers could project contamination potential for an infestation starting with a single gravid female over 30, 60, and 90 d. In addition, the amount of area covered by these spots was projected to be over 12 m2 in just 3 mo, which could greatly reduce the environmental and aesthetic quality of a home., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Cimex lectularius L. populations have been documented worldwide to be resistant to pyrethroids and neonicotinoids, insecticides that have been widely used to control bed bugs. There is an urgent need to discover new active ingredients with different modes of action to control bed bug populations. Fipronil, a phenylpyrazole that targets the GABA receptor, has been shown to be highly effective on bed bugs. However, because fipronil shares the same target site with dieldrin, we investigated the potential of fipronil resistance in bed bugs. Resistance ratios in eight North American populations and one European population ranged from 1.4- to >985-fold, with highly resistant populations on both continents. We evaluated metabolic resistance mechanisms mediated by cytochrome P450s, esterases, carboxylesterases, and glutathione S-transferases using synergists and a combination of synergists. All four detoxification enzyme classes play significant but variable roles in bed bug resistance to fipronil. Suppression of P450s and esterases with synergists eliminated resistance to fipronil in highly resistant bed bugs. Target-site insensitivity was evaluated by sequencing a fragment of the Rdl gene to detect the A302S mutation, known to confer resistance to dieldrin and fipronil in other species. All nine populations were homozygous for the wild-type genotype (susceptible phenotype). Highly resistant populations were also highly resistant to deltamethrin, suggesting that metabolic enzymes that are responsible for pyrethroid detoxification might also metabolize fipronil. It is imperative to understand the origins of fipronil resistance in the development or adoption of new active ingredients and implementation of integrated pest management programs., (© The Author(s) 2021. Published by Oxford University Press on behalf of Entomological Society of America.All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

From the 1960s to the 1980s, with the implementation of nationwide 'Four-Pest Elimination' campaigns (bed bugs, Cimex lectularius L. and Cimex hemipterus (F.) (Hemiptera: Cimicidae), were listed as one of the targeted pests), bed bug infestations were gradually eliminated in most provinces in China. However, during the last two decades, reports of bed bug infestations in the Pearl River Delta of China have shown an upward trend. Up to now, the bed bug species occurring in this area was much less frequently reported. In this study, we used both morphological and molecular methods to accurately identify the species of bed bugs collected from the cities of Guangzhou and Foshan, China. Results indicated that no significant difference was observed in the mean pronotum width-to-length ratio of Guangzhou (2.6) and Foshan (2.4) specimens; however, both were significantly lower than that of a laboratory strain C. lectularius (3.1). The genetic distances of our specimens with C. hemipterus and C. lectularius were 0-0.2% and 22.2-22.6%, respectively. On the basis of the morphological characteristics and mitochondrial DNA sequence data, it can be affirmed that bed bugs collected from Guangzhou and Foshan were C. hemipterus., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Bed bugs spend most of their lives hiding in harborages, usually in the seams of mattresses and box springs and in crevices of bed frames. For insecticidal products that target these shelters, the repellency of the products for bed bugs may influence their duration of contact. Bed bugs are known to avoid contacting surfaces treated with certain insecticides. The fungal biopesticide Aprehend contains spores of the entomopathogen, Beauveria bassiana. It is sprayed around bed frames, box springs, and furniture where bed bugs are likely to walk, which includes potential shelters. Here, I investigated the influence of a permethrin-impregnated cover, ActiveGuard, on bed bug sheltering behavior and the effectiveness of combining ActiveGuard with Aprehend. Bed bugs avoided harboring in a shelter constructed with ActiveGuard compared to a nontoxic encasement-type cover. This avoidance behavior reduced mortality induced by ActiveGuard shelters compared to forced continuous contact on the ActiveGuard cover. However, while bed bugs also avoided Aprehend-treated ActiveGuard shelters, the combined treatment induced almost complete mortality and more quickly than Aprehend-treated shelters made of the encasement-type cover. This suggests compatibility between the two integrated pest management (IPM) tools even though the bed bug's avoidance behavior would suggest otherwise. Since Aprehend is highly effective against pyrethroid-resistant bed bugs, its use would provide more effective control where bed bug populations are more resistant to the permethrin-impregnated cover., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Cimex lectularius and Cimex hemipterus are the most common species of bedbugs that infest homes. Although case reports decreased substantially by the end of the 20th century, bed bugs, and especially C. lectularius, are currently suffering a resurgence mostly attributed to insecticide resistance, inadequate pest control, and increased travel. Here, we report, to the best of our knowledge, the first molecular confirmation of C. lectularius in Central America. Specimens were obtained from an apartment located in Heredia, Costa Rica. These specimens were identified morphologically as C. lectularius. The species identification was confirmed by amplifying and sequencing fragments of the cytochrome oxidase subunit I (COI) and the 16S rRNA (16S) genes. The phylogenetic analysis showed that the sequences obtained were more closely related to a C. lectularius mitochondrial complete genome sequence from China, with similarities of 98.84% (686/694) for COI and 98.97% (387/391) for 16S. The finding of C. lectularius in Costa Rica will require further investigation in order to determine the extent of current infestations and the susceptibility to insecticides, especially due to the impact that this species can have in human health, as well as the tourism industry in the region., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Despite awareness of the mutations conferring insecticide resistance in the bed bug, Cimex lectularius L. (Hemiptera: Cimicidae), within the United States few studies address the distribution and frequency of these. Within the United States, studies have focused on collections made along the East Coast and Midwest, documenting the occurrence of two mutations (V419L and L925I) within the voltage-gated sodium channel α-subunit gene shown to be associated with knockdown resistance (kdr) to pyrethroids. Here, the distribution and frequency of the V419L and L925I site variants is reported from infestations sampled within Oklahoma and its immediately adjacent states. Additionally, the presence of a mutation previously undocumented in the United States (I935F) is noted. While novel in the United States, this mutation has previously been reported in Australian and Old World populations. No infestations were found to harbor wild-type individuals, and hence susceptible, at each of the three sites. Instead, ~21% were found to possess the resistant mutation at the L925I site (haplotype B), ~77% had mutations at both the V419L and L925I sites (haplotype C), and 2% possessed the mutation at the L936F site (haplotype Ab). The high frequency of haplotype C corresponds to previous studies in the United States, and contrasts dramatically with those of the Old World and Australia. The data presented here provide insight into the contemporary occurrence of kdr-associated insecticide resistance in the South Central United States, a region for which data have previously been absent. These data suggest that New World and Old World/Australian infestations are likely to have originated from different origins., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

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., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America.)

The newly developed fungal biopesticide Aprehend, containing spores of Beauveria bassiana, is the first biological control agent to be incorporated into management programs to control the common bed bug (Cimex lectularius L.) (Hemiptera: Cimicidae). Aprehend is sprayed as barriers where bed bugs are likely to walk and pick up spores as they search for a bloodmeal. A key application target for Aprehend is the box spring, which may be covered by encasement-type or insecticide-impregnated covers. Since some insecticides can reduce the persistence of fungal spores, we tested the efficacy and spore germination percentages of Aprehend when applied to the two types of box spring covers. We found that spore germination was about 11% lower on the permethrin-impregnated ActiveGuard cover than on the encasement-type AllerEase cover. However, bed bugs exposed for 15 min to Aprehend on the two box spring covers suffered similarly high levels of mortality irrespective of the cover material. Thus, there was no inhibitory or additive effect of the ActiveGuard cover on bed bug mortality. Lastly, overall mortality was higher if bed bugs were exposed to Aprehend-treated ActiveGuard than the ActiveGuard cover alone. Our findings indicate that if pest managers are using ActiveGuard covers in combination with Aprehend, best practice would be to use ActiveGuard on mattresses and apply Aprehend directly to the box spring or to a box spring covered by an encasement-type cover., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Most field studies on Cimex lectularius are conducted in residential or hospitality industrial settings. Cimex lectularius infestations in office settings are reported, but are rarely studied. An office environment (~875 m2) consisting of 105 cubicles or pod-seating areas with persistent C. lectularius sightings over a 2-yr period was evaluated for 90 d through intensive trapping to determine C. lectularius distribution and to eliminate the infestation. The study area was partially occupied during the study period. Two treatments of amorphous silica dust (112.5 g dust in total) were applied 29 and 57 d after the first day of monitoring. A total of 32 C. lectularius were captured by interceptors over a period of 55 d. Dry ice traps captured eight additional C. lectularius. Visual inspections identified one chair with live C. lectularius and eggs. The infestation was eliminated on 69 d after initial installation of interceptors. Spatial analysis using Ripley's K and L functions showed a high level of aggregation up to a 15 meter scale. Dispersal of C. lectularius in office settings was limited. Intensive trapping plus limited insecticide dust treatments effectively detected and eliminated C. lectularius., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The common bed bug [Cimex lectularius L. (Hemiptera: Cimicidae)] and tropical bed bug [Cimex hemipterus F. (Hemiptera: Cimicidae)] resurged in the United States and many other countries over the past decades. The need for safe and effective bed bug control products propelled the development of numerous 'green insecticides', mostly with essential oils listed as active ingredients. Various inorganic and organic oils also were used for bed bug management. However, there are no published studies on their toxicities against bed bugs. In this study, we screened 18 essential oils, three silicone oils, and paraffin oil (C5-20 paraffins) for their toxicities against bed bugs. All the oils exhibited insecticidal activity in topical assays. Their toxicities varied significantly; all of the evaluated essential oils were less effective than silicone oils and paraffin oil. The LD50 values of the most effective essential oil (blood orange), paraffin oil, and the most effective silicone oil (dodecamethylpentasiloxane) are 0.184 ± 0.018, 0.069 ± 0.012, and 0.036 ± 0.005 mg per bug, respectively. Direct spray of 1% water solution of 3-[hydroxy (polyethyleneoxy) propyl] heptamethyltrisiloxane, the only silicone oil that mixes well with water, resulted in 92% bed bug mortality after 1 d. Results of this study indicate silicone oils and paraffin oil have the potential to be used as safer alternative bed bug control materials., (© The Author(s) 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Bed bugs (Cimex lectularius L.) are now endemic in most major cities, but information regarding their basic biology is still largely based on research done over four decades ago. We investigated the effects of starvation, mating, sperm storage, and female and male age on egg production and hatch. Egg production cycles varied with the number of bloodmeals that females received. Once-mated females fed every 5 d had constant egg production for ∼75 d followed by a monotonic decline to near zero. Percentage egg hatch was high and constant, but declined after ∼30 d to near zero. To determine whether the age of the female, male, or sperm affected these patterns, we mated newly eclosed females to 60-d-old virgin males, 60-d-old mated males, or newly eclosed males. Females produced the most eggs when mated to young males, followed by old mated males, and then old virgin males; percentage hatch followed a similar pattern, suggesting that sperm stored within males for long was deficient. To examine effects of sperm stored within females, we mated newly eclosed females, starved them for 30 or 60 d, then fed them every 5 d. The 60-d starved group produced fewer eggs than the 30-d starved group, and both produced fewer eggs than young females mated to old or young males. Longer periods of sperm storage within females caused lower corresponding percentage hatch. These findings indicate egg production and hatch are governed by complex interactions among female and male age, frequency of feeding and mating, and sperm condition., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Baits are a preferred method of urban pest management. Baits enable more targeted insecticide applications with a fraction of the active ingredient used in residual sprays. Bait translocation by foragers, and consequent secondary kill of nonforagers, enhances bait effectiveness in social insects, and in other group-living species like German cockroaches (Blattella germanica L.). We investigated the potential for secondary kill in bed bugs (Cimex lectularius L.), another gregarious species, using a liquid bait. We first investigated whether blood-fed adults enhance nymph survivorship within aggregations by increasing the local relative humidity (RH) and providing fecal nutrients. Higher RH (50% and 95%) resulted in greater survivorship of first instars compared with 0% RH. Therefore, in subsequent experiments, we controlled RH to decouple its effect on nymph survivorship from effects of fecal nutrients. The presence of fed or unfed adults did not increase unfed first instar survivorship, suggesting that if nymphs ingested feces, its nutritional benefits were minimal. Nymph survivorship was unaffected by the presence of adult males fed fipronil or clothianidin, suggesting that unlike in cockroaches, highly effective insecticides might not be effective as secondary kill toxicants in bed bugs. To directly compare secondary kill in first-instar bed bugs and B. germanica, we exposed both to insecticide-laden adult B. germanica feces. Whereas first-instar B. germanica died in the presence of insecticide-laden feces, bed bugs did not. We, therefore, conclude that secondary kill with neuroactive insecticides will likely not be a significant factor in bed bug population suppression., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Bed bug, Cimex lectularius L., management in low-income, high-rise housing for the elderly and disabled can be difficult. Early detection is key to slowing their spread, and reducing management cost and time needed for control. To determine the minimum number of passive monitors needed to detect low-level bed bug infestations in this environment, we evaluated three monitors placed one, two, or four per apartment in a 3 by 3 experimental design. One sticky monitor, The Bedbug Detection System, and the two pitfall monitors, ClimbUp Insect Interceptors BG and BlackOut BedBug Detectors, were evaluated. Bed bugs were trapped by the ClimbUp Insect Interceptors BG and the BlackOut BedBug Detector in 88% and 79% of apartments, respectively, but only in 39% of the apartments monitored with The Bedbug Detection System. The Bedbug Detection System required significantly longer time to detect bed bugs than either the ClimbUp Insect Interceptor BG or the BlackOut BedBug Detector. With the less effective Bedbug Detection System data removed from analyses, detection rates ranged from 80 to 90%, with no significant differences among one, two, or four monitors per apartment. Results indicate it is especially important to include a bed placement when only placing a few monitors. Future work should compare the combination of cursory visual inspections with various monitor numbers and placements per apartment to determine the most efficient, cost-effective system that will be accepted and implemented in low-income housing., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Bed bugs, Cimex lectularius L., feed on humans, have increased exponentially in the past two decades in many major cities throughout the world, have caused intense infestations, and have become a significant health concern. Improved traps are needed to detect early infestations, to assess control programs, and for control of bed bugs. Carbon dioxide released alone or simultaneously with other attractants into three types of traps at the relatively low rate of 1 ml/min caught significantly more bed bugs than untreated controls in a 183- by 183-cm arena. This finding may enable CO2 to be used more economically in traps. Three percent ammonium bicarbonate released at a rate of ≤0.03 ml/h also caught significantly more bed bugs than untreated controls. A blend of (E)-2-hexenal and (E)-2-octenal at concentrations of 0.025% or 0.1% each and released at 0.02 ml/h attracted significantly more bed bugs than untreated controls. These findings ought to improve detection of bed bugs., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

The ability to mark individuals is a critical feature of many entomological investigations, including dispersal studies. Insect dispersal is generally investigated using mark-release-recapture techniques, whereby marked individuals are released at a known location and then captured at a measured distance. Ectoparasite dispersal has historically been challenging to study, in part because of the ethical concerns associated with releasing marked individuals. Here, we introduce the protein self-marking technique, whereby ectoparasites mark themselves in the field by feeding on the blood of an introduced host. We demonstrate the potential of this technique using laboratory-reared bed bugs (Cimex lectularius L.) that marked themselves by feeding on either rabbit or chicken blood. We then used enzyme-linked immunosorbent assays to detect host-specific blood serum proteins in bed bugs. We assessed these protein markers' ability to 1) distinctively identify marked individuals, 2) persist following multiple feedings on an alternate diet, 3) persist over time across a range of temperatures, and 4) transfer from marked to unmarked individuals. Protein markers were detectable in bed bugs before and after molting, remained detectible after multiple feedings on an alternate diet, persisted regardless of whether an individual was starved or fed on an alternate diet following original mark acquisition, and did not transfer between individuals. The duration of detectability depended on temperature. Our results suggest that protein self-marking is an effective technique for marking bed bugs and holds promise for use in dispersal studies of ectoparasitic insects., (© The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Behavioral bioassays were conducted to determine whether bed bug adults and nymphs prefer specific colored harborages. Two-choice and seven-choice behavioral color assays indicate that red (28.5%) and black (23.4%) harborages are optimal harborage choices for bed bugs. Yellow and green harborages appear to repel bed bugs. Harborage color preferences change according to gender, nutritional status, aggregation, and life stage. Female bed bugs prefer harborages with shorter wavelengths (lilac-14.5% and violet-11.5%) compared to males, whereas males prefer harborages with longer wavelengths (red-37.5% and black-32%) compared with females. The preference for orange and violet harborages is stronger when bed bugs are fed as opposed to when they are starved. Lone bed bugs (30%) prefer to be in black harborages while red harborages appear to be the optimum harborage color for bed bugs in more natural mixed aggregations (35.5%). Bed bug nymphs preferred different colored harborages at each stage of development, which is indicative of their developing eye structures and pigments. First instars showed no significant preference for any colored harborage soon after hatching. However, by the fifth instar, 27.5% of nymphs significantly preferred red and black harborages (which was a similar preference to adult bed bugs). The proportion of oviposited eggs was significantly greater under blue, red, and black harborages compared to other colored harborages tested. The use of visual cues such as specific colors offers great potential for improving bed bug monitoring tools by increasing trap captures., (© The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

We examined bed bug prevalence in 2,372 low-income apartments within 43 buildings in four New Jersey cities using a combination of resident interviews, brief visual inspections, and monitoring with Climbup Insect Interceptors. Infestation rates ranged from 3.8 to 29.5% among the buildings, with an overall infestation rate of 12.3%. Within each apartment, the bed area trapped significantly more bed bugs per trap than the sofa (or upholstered chair) area. African American residents had a proportionally higher number of bed bug infestations than white residents. Women were more likely to report bed bug bite symptoms than men. Only 68% of the residents who experienced bed bug infestations reported symptoms after being bitten (n = 475). Among those with self-reported symptoms (n = 319), the frequency of the reported symptoms was: pain 90%, itchiness 20%, welts 13%, and insomnia 8%. Fifty-nine percent of the residents (n = 539) who experienced bed bug infestations applied insecticides to control bed bugs. Climbup interceptors detected 89 ± 1% and brief visual inspections detected 72 ± 3% of the infestations. Only two out of 291 infestations were not detected by brief visual inspection or interceptors. Assuming US$50 per hour labor rate, the average per apartment cost for the building-wide bed bug monitoring protocol was US$12 per apartment. Forty-nine percent of the infestations detected by the protocol were in apartments whose residents were unaware of the bed bug activity., (© The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Insecticide sprays and dusts are used for controlling bed bugs, Cimex lectularius L. In natural environments, bed bugs have daily access to hosts after they are exposed to insecticides. The established laboratory insecticide bioassay protocols do not provide feeding after insecticide treatments, which can result in inflated mortality compared with what would be encountered in the field. We evaluated the effect of posttreatment feeding on mortality of bed bugs treated with different insecticides. None of the insecticides tested had a significant effect on the amount of blood consumed and percent feeding. The effect of posttreatment feeding on bed bug mortality varied among different insecticides. Feeding significantly reduced mortality in bed bugs exposed to deltamethrin spray, an essential oil mixture (Bed Bug Fix) spray, and diatomaceous earth dust. Feeding increased the mean survival time for bed bugs treated with chlorfenapyr spray and a spray containing an essential oil mixture (Ecoraider), but did not affect the final mortality. First instars hatched from eggs treated with chlorfenapyr liquid spray had reduced feeding compared with nymphs hatched from nontreated eggs. Those nymphs hatched from eggs treated with chlorfenapyr liquid spray and successfully fed had reduced mortality and a higher mean survival time than those without feeding. We conclude that the availability of a bloodmeal after insecticide exposure has a significant effect on bed bug mortality. Protocols for insecticide efficacy testing should consider offering a bloodmeal to the treated bed bugs within 1 to 3 d after treatment., (© The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Human bed bug infestations have dramatically increased worldwide since the mid-1990s. A similar phenomenon was also observed in Israel since 2005, when infestations were reported from all over the country. Two single nucleotide polymorphisms (V419L and L925I) in the bed bug voltage-sensitive sodium channel confer kdr-type resistance to pyrethroids. Using quantitative sequencing (QS), the resistance allele frequencies of Israeli bed bug populations from across the country were determined. Genomic DNA was extracted from samples of 12 populations of bed bugs collected from Israel and DNA fragments containing the V419L or L925I and I936F mutations sites were PCR amplified. The PCR products were analyzed by QS and the nucleotide signal ratios calculated and used to predict the resistance allele frequencies of the unknown populations. Results of the genetic analysis show that resistant nucleotide signals are highly correlated to resistance allele frequencies for both mutations. Ten of the 12 tested populations had 100% of the L925I mutation and 0% of the V419L mutation. One population was heterogeneous for the L925I mutation and had 0% of the V419L mutation and another population was heterogeneous for the V419L mutation and had 100% of the L925I mutation. I936F occurred only at low levels. These results indicate that bed bugs in Israel are genetically resistant to pyrethroids. Thus, pyrethroids should only be used for bed bug management with caution using effective application and careful monitoring procedures. Additionally, new and novel-acting insecticides and nonchemical means of controlling bed bugs should be explored., (© The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Homoplasmy, the occurrence of a single mitochondrial DNA haplotype within an individual, has been the accepted condition across most organisms in the animal kingdom. In recent years, a number of exceptions to this rule have been reported, largely due to the ease with which single nucleotide polymorphisms can be detected. Evidence of heteroplasmy-two or more mitochondrial variants within a single individual-has now been documented in a number of invertebrates; however, when present, heteroplasmy usually occurs at low frequencies both within individuals and within populations. The implications of heteroplasmy may be far reaching, both to the individual in relation to its health and fitness, and when considering the evolutionary dynamics of populations. We present novel evidence for frequent mtDNA heteroplasmy in the bed bug, Cimex lectularius L. (Hemiptera: Cimicidae). Our findings show that heteroplasmy is common, with 5 of 29 (17%) populations screened exhibiting two mitochondrial variants in a ∼1:2 ratio within each individual. We hypothesize that the mechanism underlying heteroplasmy in bed bugs is paternal leakage because some haplotypes were shared among unrelated populations and no evidence for nuclear mitochondrial DNA sequences was detected., (© The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Sublethal exposure to pesticides can alter insect behavior with potential for population-level consequences. We investigated sublethal effects of ActiveGuard, a permethrin-impregnated fabric, on feeding behavior and fecundity of bed bugs (Cimex lectularius L.) from five populations that ranged from susceptible to highly pyrethroid resistant. After exposure to ActiveGuard fabric or untreated fabric for 1 or 10 min, adult virgin female bed bugs were individually observed when offered a blood meal to determine feeding attempts and weight gain. Because bed bug feeding behavior is tightly coupled with its fecundity, all females were then mated, and the number of eggs laid and egg hatch rate were used as fecundity measures. We observed that pyrethroid-resistant and -susceptible bugs were not significantly different for all feeding and fecundity parameters. Bed bugs exposed to ActiveGuard for 10 min were significantly less likely to attempt to feed or successfully feed, and their average blood meal size was significantly smaller compared with individuals in all other groups. Independent of whether or not feeding occurred, females exposed to ActiveGuard for 10 min were significantly more likely to lay no eggs. Only a single female exposed to ActiveGuard for 10 min laid any eggs. Among the other fabric treatment-exposure time groups, there were no observable differences in egg numbers or hatch rates. Brief exposure of 10 min to ActiveGuard fabric appeared to decrease feeding and fecundity of pyrethroid-resistant and susceptible bed bugs, suggesting the potentially important role of sublethal exposure for the control of this ectoparasitic insect., (© The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Little is known about what factors influence the climbing ability of bed bugs, Cimex lectularius L. (Hemiptera: Cimicidae), in relation to the various surfaces they encounter. We examined how sex, time since last fed, and what surfaces the bed bugs were in contact with affected their climbing performance. The effects of sex and time since fed were tested by counting the number of bed bugs able to climb a 45° slope. The pulling force was recorded using an analytical balance technique that captured the sequential vertical pulling force output of bed bugs attached to various surfaces. Recently fed female bed bugs were found to have the most difficulty in climbing smooth surfaces in comparison with males. This difference can be explained by the larger weight gained from bloodmeals by female bed bugs. A variety of vertical pulling forces were observed on surfaces ranging from sandpaper to talc powder-covered glass. For surfaces not treated with talc powder, bed bugs generated the least amount of vertical pulling force from synthetically created 0.6-µm plastron surfaces. This vast range in the ability of bed bugs to grip onto various surfaces may have implications on limiting bed bugs dispersal and hitchhiking behaviors., (© The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Bed bugs Cimex lectularius L. were exposed to conidia (spores) of the entomopathogenic fungus Metarhizium anisopliae by feeding, aerosol spray, or contact with a treated surface. Feeding experiments demonstrated that bed bugs were innately susceptible to this fungus. However, only at 98% humidity were mortality rates high, regardless of whether bed bugs were sprayed with a fungal solution or contacted a treated surface. Mortality in treated bed bugs at ambient humidity did not increase when these bed bugs were kept in aggregation with other bed bugs that had recently blood fed to repletion. Based on these laboratory studies, we conclude that M. anisopliae is a poor pathogen for use in control of bed bugs, particularly at humidities that would likely be encountered under field conditions., (© 2014 Entomological Society of America.)

Detection of low-level bed bug, Cimex lectularius L. (Hemiptera: Cimicidae), infestations is essential for early intervention, confirming eradication of infestations, and reducing the spread of bed bugs. Despite the importance of detection, few effective tools and methods exist for detecting low numbers of bed bugs. Scent dogs were developed as a tool for detecting bed bugs in recent years. However, there are no data demonstrating the reliability of trained canines under natural field conditions. We evaluated the accuracy of 11 canine detection teams in naturally infested apartments. All handlers believed their dogs could detect infestations at a very high rate (≥95%). In three separate experiments, the mean (min, max) detection rate was 44 (10-100)% and mean false-positive rate was 15 (0-57)%. The false-positive rate was positively correlated with the detection rate. The probability of a bed bug infestation being detected by trained canines was not associated with the level of bed bug infestations. Four canine detection teams evaluated on multiple days were inconsistent in their ability to detect bed bugs and exhibited significant variance in accuracy of detection between inspections on different days. There was no significant relationship between the team's experience or certification status of teams and the detection rates. These data suggest that more research is needed to understand factors affecting the accuracy of canine teams for bed bug detection in naturally infested apartments., (© 2014 Entomological Society of America.)