The common bed bug Cimex lectularius synthesizes hemozoin as an essential defense against the toxic effects of heme.

The common bed bug Cimex lectularius (Linnaeus 1758) is an ectoparasite that feeds preferably on human blood, being considered an important public health issue. Blood-feeding is a challenging process for hematophagous organisms, and one of the inherent risks with this kind of diet is the liberation of high doses of free heme after the digestion of hemoglobin. In order to deal with this potent cytotoxic agent, such organisms have acquired different defense mechanisms. Here, we use UV-visible spectrophotometry and infrared spectroscopy to show that C. lectularius crystalizes free heme to form the much less dangerous compound, hemozoin. According to our results, the peak of formation of hemozoin in the intestinal contents occurred 4-5 days after the blood meal, primarily in the posterior midgut. The quantification of the rate of conversion of heme to hemozoin revealed that at the end of digestion all the heme was in the form of hemozoin. Inhibition of the synthesis of hemozoin using the anti-malarial drug quinine led to an increase in both catalase activity in the intestinal epithelium and the mortality of the bed bugs, indicating that the insects were unable to cope with the oxidative stress generated by the overload of free heme. The data presented here show for the first time how C. lectularius deals with free heme, and how the process of formation of hemozoin is essential for the survival of these insects. Since resistance to insecticides is a common feature among field populations of bed bugs, there is an urgent need to develop alternative control methods. Thus, targeting the synthesis of hemozoin emerges as a possible novel strategy to fight bed bugs.

Female bed bugs (Cimex lectularius L) anticipate the immunological consequences of traumatic insemination via feeding cues.

Not all encounters with pathogens are stochastic and insects can adjust their immune management in relation to cues associated with the likelihood of infection within a life cycle as well as across generations. In this study we show that female insects (bed bugs) up-regulate immune function in their copulatory organ in anticipation of mating by using feeding cues. Male bed bugs only mate with recently fed females and do so by traumatic insemination (TI). Consequently, there is a tight temporal correlation between female feeding and the likelihood of her being infected via TI. Females that received predictable access to food (and therefore predictable insemination and infection cycles) up-regulated induced immunity (generic antibacterial activity) in anticipation of feeding and mating. Females that received unpredictable (but the same mean periodicity) access to food did not. Females that anticipated mating-associated immune insult received measurable fitness benefits (survival and lifetime reproductive success) despite laying eggs at the same rate as females that were not able to predict these cycles. Given that mating is a time of increased likelihood of infection in many organisms, and is often associated with temporal cues such as courtship and/or feeding, we propose that anticipation of mating-associated infection in females may be more widespread than is currently evidenced.

Blood deprivation and heat stress increase mortality in bed bugs (Cimex lectularius) exposed to insect pathogenic fungi or desiccant dust.

Bed bugs (Cimex lectularius L.) have returned as a nuisance pest in the last 20 years. Different bed bug control measures in combination have not been thoroughly studied, although induction of multiple stressors may improve extermination. The effects of heat stress only, heat stress followed by exposure to insect pathogenic fungi, and heat stress followed by exposure to desiccant dust on starved and blood-fed bed bugs were investigated. Five days at 22 °C (control), 32 °C, 34 °C, or 36 °C (heat stress) did not cause mortality in adults. However, their starved first instar nymphs produced after heat stress suffered mortalities of 33%, 56% and 100%, respectively. Exposure to insect pathogenic fungi after heat stress increased the mortality of adults and their progeny compared to exposure to fungi without heat stress. The beneficial effects of heat stress were not observed in blood-fed bed bugs. Desiccant dust killed all nymphs within 2 days and all adults within 3 days regardless of previous heat stress, but survival time was prolonged by access to blood. This study highlights the advantage of combining different methods in pest management, and points to heat stress combined with blood deprivation as possible management elements to increase the control success.

Evidence for a sexually selected function of the attachment system in bedbugs Cimex lectularius (Heteroptera, Cimicidae).

Attachment to surfaces is a major aspect of an animal's interaction with the environment. Consequently, shaping of the attachment system in relation to weight load and substrate is considered to have occurred mainly by natural selection. However, sexual selection may also be important because many animals attach to their partner during mating. The two hypotheses generate opposing predictions in species where males are smaller than females. Natural selection predicts that attachment ability will scale positively with load, and hence body size, and so will be larger in females than males. Sexual selection predicts attachment forces in males will be larger than those in females, despite the males' smaller size because males benefit from uninterrupted copulation by stronger attachment to the female. We tested these predictions in the common bedbug Cimex lectularius, a species in which both sexes, as well as nymphs, regularly carry large loads: blood meals of up to 3 times their body weight. By measuring attachment forces to smooth surfaces and analysing in situ fixed copulating pairs and the morphology of attachment devices, we show that: (i) males generate twice the attachment force of females, despite weighing 15% less; (ii) males adhere to females during copulation using hairy tibial adhesive pads; (iii) there are more setae, and more setae per unit area, in the pads of males than in those of females but there is no difference in the shape of the tarsal setae; and (iv) there is an absence of hairy tibial attachment pads and a low attachment force in nymphs. These results are consistent with a sexually selected function of attachment in bedbugs. Controlling sperm transfer and mate guarding by attaching to females during copulation may also shape the evolution of male attachment structures in other species. More generally, we hypothesise the existence of an arms race in terms of male attachment structures and female counterparts to impede attachment, which may result in a similar evolutionary diversification to male genitalia.

Discrimination between lineage-specific shelters by bat- and human-associated bed bugs does not constitute a stable reproductive barrier.

The common bed bug Cimex lectularius, has been recently shown to constitute two host races, which are likely in the course of incipient speciation. The human-associated lineage splits from the ancestral bat-associated species deep in the history of modern humans, likely even prior to the Neolithic Period and establishment of the first permanent human settlements. Hybridization experiments between these two lineages show that post-mating reproductive barriers are incomplete due to local variation. As mating takes place in off-host refugia marked by aggregation semiochemicals, the present investigation tested the hypothesis that bed bugs use these semiochemicals to differentiate between refugia marked by bat- and human-associated bed bugs; this would constitute a pre-copulation isolation mechanism. The preference for lineage-specific odors was tested using artificial shelters conditioned by a group of either male or female bed bugs. Adult males were assayed individually in four-choice assays that included two clean unconditioned control shelters. In most assays, bed bugs preferred to rest in conditioned shelters, with no apparent fidelity to shelters conditioned by their specific lineage. However, 51 % of the bat-associated males preferred unconditioned shelters over female-conditioned shelters of either lineage. Thus, bed bugs show no preferences for lineage-specific shelters, strongly suggesting that semiochemicals associated with shelters alone do not function in reproductive isolation.

[The common bed bug (Cimex lectularius) - biology, medical relevance, possibilities for the detection and control]. / Stenice domácí (Cimex lectularius) - biologie, zdravotnický význam, moznosti detekce a hubení.

Bed bugs have become a major concern worldwide in the 21st century and are therefore intensively investigated. The new findings not only extend the knowledge of their biology, medical relevance, and causes of the resurgence, but also can be used in bed bug management. A brief overview is provided of some of the most important research results and opinions, published in the last few years in prestigious international journals.

Feel the heat: activation, orientation and feeding responses of bed bugs to targets at different temperatures.

Host location in bed bugs is poorly understood. Of the primary host-associated cues known to attract bed bugs - CO2, odors, heat - heat has received little attention as an independent stimulus. We evaluated the effects of target temperatures ranging from 23 to 48°C on bed bug activation, orientation and feeding. Activation and orientation responses were assessed using a heated target in a circular arena. All targets heated above ambient temperature activated bed bugs (initiated movement) and elicited oriented movement toward the target, with higher temperatures generally resulting in faster activation and orientation. The distance over which bed bugs could orient toward a heat source was measured using a 2-choice T-maze assay. Positive thermotaxis was limited to distances <3 cm. Bed bug feeding responses on an artificial feeding system increased with feeder temperature up to 38 and 43°C, and declined precipitously at 48°C. In addition, bed bugs responded to the relative difference between ambient and feeder temperatures. These results highlight the wide range of temperatures that elicit activation, orientation and feeding responses in bed bugs. In contrast, the ability of bed bugs to correctly orient towards a heated target, independently of other cues, is limited to very short distances (<3 cm). Finally, bed bug feeding is shown to be relative to ambient temperature, not an absolute response to feeder blood temperature.

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 (λmax 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 (λmax 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.

Estimating the critical thermal maximum (CTmax) of bed bugs, Cimex lectularius: Comparing thermolimit respirometry with traditional visual methods.

Evaluating the critical thermal maximum (CTmax) in insects has provided a number of challenges. Visual observations of endpoints (onset of spasms, loss of righting response, etc.) can be difficult to measure consistently, especially with smaller insects. To resolve this problem, Lighton and Turner (2004) developed a new technique: thermolimit respirometry (TLR). TLR combines real time measurements of both metabolism (V·CO2) and activity to provide two independent, objective measures of CTmax. However, several questions still remain regarding the precision of TLR and how accurate it is in relation to traditional methods. Therefore, we evaluated CTmax of bed bugs using both traditional (visual) methods and TLR at three important metabolic periods following feeding (1d, 9d, and 21d). Both methods provided similar estimates of CTmax, although traditional methods produced consistently lower values (0.7-1°C lower than TLR). Despite similar levels of precision, TLR provided a more complete profile of thermal tolerance, describing changes in metabolism and activity leading up to the CTmax, not available through traditional methods. In addition, feeding status had a significant effect on bed bug CTmax, with bed bugs starved 9d (45.19[±0.20]°C) having the greatest thermal tolerance, followed by bed bugs starved 1d (44.64[±0.28]°C), and finally bed bugs starved 21d (44.12[±0.28]°C). Accuracy of traditional visual methods in relation to TLR is highly dependent on the selected endpoint; however, when performed correctly, both methods provide precise, accurate, and reliable estimations of CTmax.

Posttreatment Feeding Affects Mortality of Bed Bugs (Hemiptera: Cimicidae) Exposed to Insecticides.

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.

Climbing Ability of the Common Bed Bug (Hemiptera: Cimicidae).

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.

Status of Urban Bed Bug Infestations in Southern China: An Analysis of Pest Control Service Records in Shenzhen in 2012 and Dongguan in 2013.

The recent resurgence of bed bugs (Cimex spp.) in many developed countries has drawn increasing attention worldwide. The status of urban bed bug infestations were investigated in Shenzhen and Dongguan, two major cities in southern Guangdong Province of southern China, based on pest control service records from two different companies (one during 2012 and another during 2013). The results showed that Shenzhen and Dongguan have a severe problem with bed bug infestations: the control of bed bugs is a constant concern, except during the winter. In Shenzhen, a similar number of premises were treated for bed bugs in central business districts and suburban districts. However, in Dongguan, more premises were treated for bed bugs in suburban districts than in central business districts. The treatment rate for worker sleeping quarters, apartments, hotel, and private houses in Shenzhen was 53.8, 43.0, 1.9, and 1.3%, respectively. The percentage of treated rooms was 56.1% for worker sleeping quarters and 91.1% for apartments. In Dongguan, the treatment rate for worker sleeping quarters, apartments, hotel, and private houses was 90.0, 10.0, 0.0, and 0.0%, respectively.

Sublethal Effects of ActiveGuard Exposure on Feeding Behavior and Fecundity of the Bed Bug (Hemiptera: Cimicidae).

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.

Reproduction barrier between two lineages of bed bug (Cimex lectularius) (Heteroptera: Cimicidae).

Populations of bed bugs, Cimex lectularius, have increased in recent years spreading into numerous urban areas across the Western world and making them an increasingly important pest of the twenty-first century. Research into hybridization within and between different lineages of bed bugs can help us to understand processes of micro- and macro-evolution in these ectoparasites and may inform the control of this pest species. Hybridization experiments between two host lineages of bed bug (C. lectularius) from Central Europe (Czech Republic), those associated with humans and those with bats, were conducted under laboratory conditions. Number of eggs and early instars were compared between crosses of mixed host lineages (interspecific mating) with pairs from the same host lineage, those from the same locality and same lineage from different localities (intraspecific mating). While crosses within host lineages resulted in egg production and later instars, crosses between different host lineages were unsuccessful, although of the mated females possessed sperm in their mesospermaleges and/or seminal conceptacles. These crosses did not even result in egg production. Moreover, in the mixed lineage crosses, mortality rates in adults were higher (51 and 50% higher in bat and human lineage, respectively) than in those animals from the same lineage. Survival of adults was in pairs from the same locality slightly higher than in pairs from different localities and differed statistically. These results support the existence of post-mating barriers and show reproductive isolation between two lineages of C. lectularius. Bat and human host adaptations can promote evolving of such barriers and can be product of alloxenic speciation.

Blood constituents as phagostimulants for the bed bug Cimex lectularius L.

Many hematophagous arthropods are stimulated by blood constituents to initiate feeding. We used a membrane-based feeding system to identify chemicals that stimulate acceptance and engorgement responses in various life stages of bed bugs. Water was fortified with a variety of compounds (e.g. salts, amino acids, vitamins, nucleotides, cholesterol and fatty acids) in these bioassays. ATP was the most effective phagostimulant in adults and nymphs, resulting in >70% of bed bugs fully engorging. Addition of NaCl to low ATP solutions that alone elicited <50% engorgement significantly enhanced feeding responses of bed bugs. A comparison of feeding responses with solutions of various adenine nucleotides showed that ATP was more stimulatory than ADP, which was more effective than AMP. Feeding assays with physiological levels of other blood constituents such as d-glucose, albumin, globulin, cholesterol and mixtures of vitamins and amino acids did not stimulate engorgement, suggesting that adenine nucleotides are the most important feeding stimulants in bed bugs. Identification of phagostimulants for bed bugs will contribute towards the development of artificial diets for rearing purposes, as well as for the development of alternative methods to eliminate bed bug infestations.

Bed bugs are back and also arriving is the southernmost record of Cimex lectularius (Heteroptera: Cimicidae) in South America.

The arrival and establishment of the bed bug Cimex lectularius L., 1755 in Magallanes Region (Chile) is reported. The present record is the southernmost for this species in America. The circumstances that provided for the species reaching this southern locality are discussed.

Bed bugs, their blood sources and life history parameters: a comparison of artificial and natural feeding.

A blood-feeding system that utilizes a small amount of whole heparinized human blood in parafilm bags is described in detail, and similarities and differences between artificially fed and naturally rodent-fed bed bugs (Hemiptera: Cimicidae) are discussed. Blood with high levels of heparin (10%) was unsuitable for artificial colony rearing, whereas bed bugs fed on 1% heparinized blood and those that naturally ingested rat blood completed their lifecycle with similar stage structures over time, with no significant differences in mortality. No differences in feeding efficiency or fertility were found in a direct comparison of bed bugs maintained under each of these two treatments, but analysis of the full lifecycle revealed that artificially fed bed bugs became significantly smaller and laid fewer eggs than rodent-fed bed bugs. The level of membrane stretching regulated the number of bed bugs that fed. When the membrane was stretched to twice its length and width, 96% of bed bugs successfully fed through the parafilm. Whole heparinized blood that was stored at 6 °C for ≥ 14 days failed to retain its nutritional value and the amount of blood consumed and number of consecutive moults were significantly reduced.

Drinking a hot blood meal elicits a protective heat shock response in mosquitoes.

The mosquito's body temperature increases dramatically when it takes a blood meal from a warm-blooded, vertebrate host. By using the yellow fever mosquito, Aedes aegypti, we demonstrate that this boost in temperature following a blood meal prompts the synthesis of heat shock protein 70 (Hsp70). This response, elicited by the temperature of the blood meal, is most robust in the mosquito's midgut. When RNA interference is used to suppress expression of hsp70, protein digestion of the blood meal is impaired, leading to production of fewer eggs. We propose that Hsp70 protects the mosquito midgut from the temperature stress incurred by drinking a hot blood meal. Similar increases in hsp70 were documented immediately after blood feeding in two other mosquitoes (Culex pipiens and Anopheles gambiae) and the bed bug, Cimex lectularius, suggesting that this is a common protective response in blood-feeding arthropods.

Effect of feeding status on mortality response of adult bed bugs (Hemiptera: Cimicidae) to some insecticide products.

Fresh and aged residual deposits of several insecticide products were tested against bed bug adults to determine if a recent bloodmeal affected their mortality response to the residues. The bed bugs with a recent bloodmeal survived significantly longer compared with the unfed ones on their exposure to fresh or aged residual deposits of chlorfenapyr and aged residual deposits of deltamethrin on a wooden substrate. Even though the survival time of fed bed bugs was significantly longer than that of unfed ones on their exposure to fresh residue of deltamethrin and aged residue of desiccant pyrethrin dust, these treatments resulted in similarly high final mortalities regardless of feeding status of the insects. Mortality responses of fed and unfed bed bugs were similar to fresh or aged residual deposits of imidacloprid + cyfluthrin combination and fresh residual deposits of desiccant pyrethrin dust. Topical application assays indicated that a recent bloodmeal significantly increased the bed bug's survival time for chlorfenapyr, but not for deltamethrin. Pyrethroid-resistant bed bugs also showed a similar increase in their survival time for chlorfenapyr after a bloodmeal. The comparison of mortality responses between fed and unfed bed bugs treated with similar amount of chlorfenapyr per fresh body weight indicated that increased body mass was not the primary cause for this bloodmeal-induced tolerance increase for chlorfenapyr. Because the surviving bed bugs can continue ovipositing, the effectiveness of chlorfenapyr residual deposits in bed bug harborages could be significantly affected by the feeding status of the adult bed bug populations.

Aggregation responses of Cimex hemipterus F. to semiochemicals identified from their excreta.

BACKGROUND & OBJECTIVES: Bedbug Cimex hemipterus lives in cracks and crevices, and shows aggregation activity in the harbourage containing its fecal matter. Limited information is available on the chemicals influencing this aggregation behaviour in C. hemipterus. Possible components responsible for aggregation, if identified and evaluated can be helpful in designing bedbug management. METHODS: Study on the isolation and identification of chemicals from the excreta extract responsible for aggregation has been carried by GC-MS. The compounds identified were evaluated at different concentrations to C. hemipterus and their attraction index was determined. RESULTS: The GC-MS analysis of the excreta extract resulted in identification of 33 different types of semiochemicals such as esters, carboxylic acids, alcohols, aldehydes, ketones and other hydrocarbons. Among these 33 compounds of various groups, the carboxylic acid (E)-2-hexenoic acid and the aldehyde (E)-2-hexenal found to elicit highest positive aggregation behavioural response in all the stages followed by hexenal. INTERPRETATION & CONCLUSION: The fecal matter influence the aggregation behaviour in C. hemipterus. (E)-2-hexenoic acid, hexanoic acid, (E)-2-hexenal and hexanal found to exhibit aggregation in various stages of bedbug life cycle. These chemical components can be further exploited for designing and development tool for management of bedbugs.