Evidence for male- and juvenile-specific contact pheromones of the common bed bug Cimex lectularius

Males and females of the common bed bug, Cimex lectularius L. (Heteroptera: Cimicidae), have been shown to produce and respond to an aggregation pheromone. We tested whether juvenile C. lectularius also produce and respond to aggregation pheromone, and whether the pheromone is perceived by contact chemoreception. In dual‐choice laboratory experiments, juveniles, but not males or females, preferred juvenile‐exposed paper discs to control discs. Unlike juveniles, males and females preferred male‐exposed paper discs to control discs. Neither juveniles, males, nor females preferred female‐exposed discs to control discs. When test stimuli were inaccessible, C. lectularius failed to show any preference. Male‐ and juvenile‐specific contact pheromones may have contrasting functions of marking shelters as safe refugia for development and growth (juveniles) or mate encounter (adults), but result in the same phenomenon, the aggregation of conspecifics.     

Opportunities for female choice in the bed bug Cimex lectularius

Bed bugs are cited as exemplars of sexual conflict because mating can only occur via traumatic insemination. However, past antagonistic coevolution between the sexes does not necessarily preclude current female choice. Here, we investigate opportunities for precopulatory female choice in bed bugs. We examined whether females seek out mating opportunities when they gain the most benefit: when females are virgin and/or have recently fed. But, we found that female mating and feeding status had little effect on female attraction to males and male odor. To determine whether females approach male harborages (home crevices) to seek matings in nature, we investigated where matings occurred among unfamiliar pairs of bed bugs. We found that, despite female attraction to male odor, matings were most likely to take place in the female's harborage rather than the male's harborage. We also examined the effect of feeding on male and female ability to mate. Whereas previous research reported that engorgement impaired female ability to refuse matings, we found that male feeding status had a larger effect on the success of mating encounters than female feeding status. Fed males had poor mating success, suggesting that males may be faced with a trade‐off between mating and feeding.     

Host association drives genetic divergence in the bed bug,Cimex lectularius

Genetic differentiation may exist among sympatric populations of a species due to long‐term associations with alternative hosts (i.e. host‐associated differentiation). While host‐associated differentiation has been documented in several phytophagus insects, there are far fewer cases known in animal parasites. The bed bug, Cimex lectularius, a wingless insect, represents a potential model organism for elucidating the processes involved in host‐associated differentiation in animal parasites with relatively limited mobility. In conjunction with the expansion of modern humans from Africa into Eurasia, it has been speculated that bed bugs extended their host range from bats to humans in their shared cave domiciles throughout Eurasia. C. lectularius that associate with humans have a cosmopolitan distribution, whereas those associated with bats occur across Europe, often in human‐built structures. We assessed genetic structure and gene flow within and among populations collected in association with each host using mtDNA, microsatellite loci and knock‐down resistance gene variants. Both nuclear and mitochondrial data support a lack of significant contemporary gene flow between host‐specific populations. Within locations human‐associated bed bug populations exhibit limited genetic diversity and elevated levels of inbreeding, likely due to human‐mediated movement, infrequent additional introduction events per infestation, and pest control. In contrast, populations within bat roosts exhibit higher genetic diversity and lower levels of relatedness, suggesting populations are stable with temporal fluctuations due to host dispersal and bug mortality. In concert with previously published evidence of morphological and behavioural differentiation, the genetic data presented here suggest C. lectularius is currently undergoing lineage divergence through host association.     

The antennal sensilla of the praying mantis Tenodera aridifolia: A new flagellar partition based on the antennal macro-, micro- and ultrastructures

In insects, the antenna consists of a scapus, a pedicellus, and a flagellum comprising many segments (flagellomeres). These segments possess many morphological types of sensory organs (sensilla) to process multimodal sensory information. We observed the sensilla on flagellomeres in praying mantis (Tenodera aridifolia) with both scanning and transmission electron microscopes. We classified the sensilla into six types: chaetic, campaniform, coelocapitular, basiconic, trichoid and grooved peg sensilla, and inferred their presumptive functions on the basis of their external and internal structures. In addition, based on their distribution, we newly divided the flagellum into 6 distinct parts. This new division leads to a better understanding about the sexual dimorphism and the antennal development in the mantises. The sexual difference in distribution of the grooved peg sensilla suggests that this type of sensilla may play a role in sex-pheromone detection in mantis, which is a rare case of double-walled sensilla mediating this function.     

Water loss and metabolic activity in bed bug eggs (Cimex lectularius)

Few studies have evaluated water loss and respiratory activity of insect eggs, particularly insects that are known to live within indoor environments. The present study quantifies water loss and respiratory activity for the eggs of a re‐emerging indoor pest of human environments Cimex lectularius (L.). Water loss is measured gravimetrically and calculated as a function of chorion permeability. For these studies, bed bug eggs are placed at 0% relative humidity and repeatedly weighed over 48 h. Temperature effects and bed bug strain differences on the standard metabolic rate (SMR) and respiratory quotient are measured using closed system respirometry. The SMR (; mL g^?1 h^?1) is measured for two field strain bed bugs and compared with a laboratory strain held at one temperature (25 °C). The standard metabolic rate is measured for Harlan (laboratory) strain bed bug eggs at six different temperatures (15, 20, 25, 30, 35 and 39 °C). Total water loss is not significantly different between all three strains. However, water loss across the chorion (chorion permeability) is significantly different between the Harlan laboratory strain and the two field collected strains. Standard metabolic rates for Harlan (laboratory) strain bed bug eggs increase with temperatures from 15 to 35 °C but decline at 39 °C. Overall, the Harlan bed bug eggs have the largest standard metabolic rates (0.18 ± 0.05 mL g^?1 h^?1) compared with the Epic Center strain eggs (0.14 ± 0.03 mL g^?1 h^?1) and Richmond strain eggs (0.16 ± 0.04 mL g^?1 h^?1), although this difference is not significant.     

有瓣蝇类触角感受器形态与功能研究进展

有瓣蝇类隶属于昆虫纲双翅目,其物种多样性高,适应能力强,生态类型丰富,与人类关系密切,是开展昆虫适应演化研究的理想类群。触角是有瓣蝇类最重要的嗅觉感受器官,在其精准寻找食源,高效完成交配、产卵等生活史环节中都起着获取外界信息的关键作用。目前已有大量对于有瓣蝇类触角感受器的研究,但这些研究对触角感受器形态名词的使用存在诸多差异、混乱和歧义,使得不同研究间难以相互参考。本文统一了之前研究中有瓣蝇类触角上常见的各类感受结构的不同名词;并结合其它昆虫类群的相关研究,综述了各类感受器在形态和功能方面的研究进展;探讨了该领域中尚待解决的问题。     

Supernumerary chromosomes in the human bed bug,Cimex lectularius Linn. (Cimicidae:Hemiptera)

The chromosome complement in the human bed bug, Cimex lectularius Linn., is 26+X₁X₂Y in the male and 26+X₁X₁X₂X₂ in the female. However, a population from Cairo, Egypt has 4 supernumerary X chromosomes. In the hybrid between the Berkeley population (with no supernumeraries) and the Cairo population (with 4 supernumeraries), the behavior of supernumeraries was observed during embryogenesis and oogenesis as well as spermatogenesis.In embryogenesis the transmission of supernumeraries was quite regular. However, one chromosome may sometimes be eliminated early in the germ line. This abnormality could induce the variations in chromosome number encountered in later stages. In the first meiotic division, some of the supernumeraries were nondisjunctional. Moreover, in the second division, some supernumeraries were eliminated. These results show that there is a tendency towards a decrease in the number of supernumeraries in the hybrids.Although the supernumeraries behave like X chromosomes, they seem not to be important for sex determination and appear to be largely or entirely inert genetically. Supernumeraries in the bed bug originate from small fragments caused by structural rearrangement. They are increased by an accumulation mechanism. Supernumeraries in the bed bug appear to be of relatively recent origin. The phylogenetic sequence in their development was probably from none to a stabilized number of four in the Old World. Then the supernumeraries were lost in two specialized lines, Cimex columbarius Jenyns on domestic birds in Europe and Cimex lectularius Linn. on man in the Western Hemisphere.This study was carried out under U.S. Public Health Service Grant (GM-13197).     

RNA interference of NADPH-cytochrome P450 reductase results in reduced insecticide resistance in the bed bug, Cimex lectularius

Background

NADPH-cytochrome P450 reductase (CPR) plays a central role in cytochrome P450 action. The genes coding for P450s are not yet fully identified in the bed bug, Cimex lectularius. Hence, we decided to clone cDNA and knockdown the expression of the gene coding for CPR which is suggested to be required for the function of all P450s to determine whether or not P450s are involved in resistance of bed bugs to insecticides.

Methodology/Principal Findings

The full length Cimex lectularius CPR (ClCPR) cDNA was isolated from a deltamethrin resistant bed bug population (CIN-1) using a combined PCR strategy. Bioinformatics and in silico modeling were employed to identify three conserved binding domains (FMN, FAD, NADP), a FAD binding motif, and the catalytic residues. The critical amino acids involved in FMN, FAD, NADP binding and their putative functions were also analyzed. No signal peptide but a membrane anchor domain with 21 amino acids which facilitates the localization of ClCPR on the endoplasmic reticulum was identified in ClCPR protein. Phylogenetic analysis showed that ClCPR is closer to the CPR from the body louse, Pediculus humanus corporis than to the CPRs from the other insect species studied. The ClCPR gene was ubiquitously expressed in all tissues tested but showed an increase in expression as immature stages develop into adults. We exploited the traumatic insemination mechanism of bed bugs to inject dsRNA and successfully knockdown the expression of the gene coding for ClCPR. Suppression of the ClCPR expression increased susceptibility to deltamethrin in resistant populations but not in the susceptible population of bed bugs.

Conclusions/Significance

These data suggest that P450-mediated metabolic detoxification may serve as one of the resistance mechanisms in bed bugs.     

Effects of elevated temperature on the mycetome and symbiotes of the bed bug Cimex lectularius (Heteroptera)

High temperature treatment significantly reduced the reproductive rate of Cimex lectularius and simultaneously rendered their mycetomes nearly symbiote-free. The possibility that the symbiotes may affect host reproductive potential is discussed.     

Vision in the common bed bug Cimex lectularius L. (Hemiptera: Cimicidae): eye morphology and spectral sensitivity

Bed bugs as pests of public health importance recently experienced a resurgence in populations throughout the U.S. and other countries. Consequently, recent research efforts have focused on improving understanding of bed bug physiology and behaviour to improve management. While few studies have investigated the visual capabilities of bed bugs, the present study focused specifically on eye morphology and spectral sensitivity. A 3‐D imaging technique was used to document bed bug eye morphology from the first instar through adult and revealed morphological characteristics that differentiate the common bed bug from the tropical bed bug as well as sex‐specific differences. Electrophysiological measurements were used to evaluate the spectral sensitivity of adult bed bugs. Male bed bugs were more responsive than females at some wavelengths. Electrophysiological studies provided evidence for at least one photoreceptor with a spectral sensitivity curve peak in the green (λ_max 520 nm) region of the spectrum. The broadened long wavelength portion of the spectral sensitivity curve may potentially indicate another photoreceptor in the yellow–green (λ_max 550 nm) portion of the spectrum or screening pigments. Understanding more about bed bug visual biology is vital for designing traps, which are an important component of integrated bed bug management.     

Sexual dimorphism of arrestment and gregariousness in the bed bug (Cimex lectularius) in response to cuticular extracts from nymphal exuviae

Releaser pheromones have direct behavioural effects to arrest, attract or disperse insects, whereas interactions within groups of social insects are often influenced by primer pheromones. The behaviour of insects displaying intermediate levels of sociality is largely unexplored in this context. In the present study, both the gregariousness and arrestment (settling near the odour source) of bed bugs Cimex lectularius L. (Hemiptera: Cimicidae) in response to conspecific exuvial extracts are described. Adult males are arrested on filter papers with extracts derived from exuviae of fifth‐instar nymphs. Adult females and nymphs display no significant evidence for such behaviour. Adults of both sexes show no preference for extracts of male versus female fifth‐instar exuviae. Arrestment of adult males does not occur on papers treated with fourth‐instar exuvial extracts. Because the insects are assayed behaviourally in groups, an index is calculated describing how far bugs are away from being located independently of one another, as a measure of gregariousness. Adult males have lower values for this index (i.e. locations are closer to independence). Adult females, nymph cohorts and mixed age groups all have higher values for this index, which tend to increase over time. Females exhibit a clear increasing dose‐dependent relationship for this index. It is concluded that the extracts of fifth‐instar nymphal exuvia arrest males on refuges that possess the odour source. However, gregariousness is induced in females, without evidence of a tendency to assemble near the odour source.     

Sensitivity to bites by the bedbug, Cimex lectularius

Abstract.  Bedbugs are a public health problem and can cause significant economic losses, but little is known about the effects of bites on humans. We reviewed case reports and published papers on bedbug bites to assess the empirical basis of the commonly cited figure that only ∼ 80% of the population are sensitive to bedbug bites. We found the sensitivity estimate to be based on only one study carried out 80 years ago. However, this study did not account for the now well-established fact that only repeated exposure to external allergens leads to skin reactions. In our sample, 18 of 19 persons showed a skin reaction after bedbug exposure, but in most cases only after repeated controlled exposure. With repeated exposure, the latency between bite and skin reactions decreased from ∼ 10 days to a few seconds. Our results are relevant for the hospitality industry, where apparently increasing infestation rates are likely to lead to an increase in the number of tourists and hotel employees exposed to bedbugs. Medical and public health professionals may expect to see an increase in the prevalence of people with bedbug bite sensitivity. The significance of the delayed reaction time of skin to bites may also have implications in litigation cases where people seek compensation.     

Structure of sensilla, olfactory perception, and behaviour of the bedbug, Cimex lectularius, in response to its alarm pheromone

When deprived of the terminal antennal segments, male and female bedbugs failed to respond to their alarm pheromone and to their assembling scent. Trans-oct-2-en-1-al or trans-hex-2-en-1-al, being the major constituents of the former, induce in adults and larvae of Cimex lectularius a typical alarm behaviour resulting in dispersal of assembled bedbugs; the rapidity of escape depends on the aldehyde concentration in the air. The behavioural threshold for adults is about 9×10¹⁴ molecules of trans-oct-2-en-1-al or 6×10¹⁵ molecules of trans-hex-2-en-1-al per ml air.The distal part of the terminal antennal segment of C. lectularius reveals the following sensilla: bristles (type A1), immersed cones (type B1), plates (type B2), grooved pegs (type C), smooth pegs (type D), hairs with even (type E1), and uneven wall thickness (type E2). The number and distribution of these sensilla is relatively constant and similar in both sexes, but differs slightly in neonate larvae. The pegs and hairs of types C, D, E1 and E2 were shown to have porous walls, a prerequisite for olfactory function.Receptor potentials were recorded from olfactory sensilla of types E1 and E2 after stimulation with trans-hex-2-en-1-al and trans-oct-2-en-1-al. The minimal concentration of trans-hex-2-en-1-al evoking a receptor potential is about 2×10¹⁰ molecules per ml air. The above olfactory sensilla were found to respond also to hexan-1-al, but almost no responses to pentan-1-al, butan-1-al, trans-hex-2-ene, and trans-oct-2-ene were observed. A minimum chain length of six carbons atoms and a terminal carbonyl group are molecular prerequisites for optimal odorant activity, while the presence of a Δ²-double bond is not essential for stimulation of the alarm pheromone receptors of the bedbug.     

Responses of the bed bug,Cimex lectularius,to temperature extremes and dehydration: levels of tolerance,rapid cold hardening and expression of heat shock proteins

This study of the bed bug, Cimex lectularius, examines tolerance of adult females to extremes in temperature and loss of body water. Although the supercooling point (SCP) of the bed bugs was approximately −20°C, all were killed by a direct 1 h exposure to −16°C. Thus, this species cannot tolerate freezing and is killed at temperatures well above its SCP. Neither cold acclimation at 4°C for 2 weeks nor dehydration (15% loss of water content) enhanced cold tolerance. However, bed bugs have the capacity for rapid cold hardening, i.e. a 1‐h exposure to 0°C improved their subsequent tolerance of −14 and −16°C. In response to heat stress, fewer than 20% of the bugs survived a 1‐h exposure to 46°C, and nearly all were killed at 48°C. Dehydration, heat acclimation at 30°C for 2 weeks and rapid heat hardening at 37°C for 1 h all failed to improve heat tolerance. Expression of the mRNAs encoding two heat shock proteins (Hsps), Hsp70 and Hsp90, was elevated in response to heat stress, cold stress and during dehydration and rehydration. The response of Hsp90 was more pronounced than that of Hsp70 during dehydration and rehydration. Our results define the tolerance limits for bed bugs to these commonly encountered stresses of temperature and low humidity and indicate a role for Hsps in responding to these stresses.     

青杨脊虎天牛触角主要感器的超微结构及其分布

利用扫描电镜观察青杨脊虎天牛Xylotrechusrusticus L.触角感器的类型和分布。结果表明,在青杨脊虎天牛触角上共有7类16种常见的感器类型:4种锥形感器、1种凹槽钉形感器、5种刺形感器、1种芽形感器、1种剑形感器、1种齿形感器、1种钩形感器、3种指形感器。根据这些感器的分布、形态特征结合已有的文献描述,分析各感器可能具有的功能。     

Reemergence of the bedbug Cimex lectularius in Seoul, Korea

A healthy 30-yr-old woman carrying an insect that had been caught in her living room visited the International Clinic at Severance Hospital, Seoul, in December 2007. The insect she brought was identified to be a nymph of a bedbug, Cimex lectularius, and her skin rashes looked typical bedbug''s bites. Her apartment was investigated, and a dead body of a bedbug, cast skins, and hatched eggs were found in her rooms and neighbors'' rooms in the same building. She was living in that apartment in Seoul for 9 months since she had moved from New Jersey, USA. We assume that the bedbugs were introduced from abroad, since there had been no report on bedbugs in Seoul for more than 2 decades at least. This is a report of a reemergence of the common bedbug, C. lectularius in Seoul, Korea.     

The role of contact chemoreception in the host location process of an egg parasitoid

Taste allows insects to detect palatable or toxic foods, identify a mate, and select appropriate oviposition sites. The gustatory system strongly contributes to the survival and reproductive success of many species, yet it is rarely studied in insect parasitoids. In order to locate and assess a host in which they will lay their eggs, female wasps actively search for chemical cues using their sensory organs present mainly on the antennae. In this paper, we studied the role of antennal taste sensilla chaetica in the perception of contact semiochemicals in Trissolcus brochymenae (Hymenoptera: Platygastridae), an egg parasitoid of the brassicaceae pest Murgantia histrionica (Heteroptera: Pentatomidae). Methanolic extracts obtained from male and female hosts elicited action potentials in taste neurons housed in antennal sensilla chaetica, indicating that these sensilla are involved in the perception of non volatile host kairomones. In behavioural assays, wasp females displayed an intense searching behaviour in open arenas treated with host extracts, thus confirming that these kairomones are soluble in polar solvents. We further investigated the extracts by Gas Chromatography-Mass Spectrometry (GC-MS) and found that they contain several compounds which are good candidates for these contact kairomones. This study contributes to better understanding contact chemoreception in egg parasitoids and identifying gustatory receptor neurons involved in the host location process.