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The family Cimicidae comprises ectoparasites feeding exclusively on the blood of endothermic animals. Cimicid swallow bugs specifically target swallow birds (Hirundinidae) and their nestlings in infested nests. Bugs of the genus Oeciacus are commonly found in mud nests of swallows and martins, while they rarely visit the homes of humans. Although-unlike other cimicid species-the house martin bug Oeciacus hirundinis has never been reported as a vector of zoonotic pathogens, its possible role in arbovirus circulation in continental Europe is unclear. Samples of O. hirundinis were therefore collected from abandoned house martin (Delichon urbicum) nests in southern Moravia (Czech Republic) during the 2021/2022 winter season and checked for alpha-, flavi- and bunyaviruses by RT-PCR. Of a total of 96 pools consisting of three adult bugs each, one pool tested positive for Usutu virus (USUV)-RNA. Phylogenetic analysis showed that the virus strain was closely related to Italian and some Central European strains and corresponded to USUV lineage 5. The detection of USUV in O. hirundinis during wintertime in the absence of swallows raises the question for a possible role of this avian ectoparasite in virus overwintering in Europe.

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BACKGROUND: Bed bugs, Cimex lectularius, form day-time aggregations from which they depart at night to feed on human blood. Obtaining an initial blood meal is a critical step in the development of first instars. Previous research had shown that first instars had greater success in obtaining this essential meal when in the presence of adults than when they were alone. METHODS: Feeding by bed bugs was tested in upright vertical cylindrical chambers fitted with a paper ramp to aid in climbing toward a blood feeder suspended across the upper end of the cylinder. Feeding success by the first instars was tested when they were alone in the chamber or when they were in the presence of adult females, males, or both together.  RESULTS: The mean proportions of the first instars that fed were significantly higher when they were confined with adults of both sexes or adult females than when they were confined alone or with males. Feeding by adult males was also enhanced by confinement with females. When first instars and adult females were confined together, the mean duration before first instars began feeding was longer than for females. There was no difference in feeding success by first instars confined with their mothers or nonmothers. CONCLUSIONS: Elevated feeding by first instars and adult males in the presence of females may be adaptive traits that enhance fitness. First instars must feed to avoid dehydration and starvation and to obtain resources needed for development. Adult males would benefit not only by increased feeding success but also by greater likelihood of finding a recently engorged female with which to mate. The lack of any difference in feeding success of first instars in the presence of their mothers or nonmothers argues against parental care in this species.

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Cimicidae are well-known for traumatic insemination, and males pierce females with their parameres and transfer sperm through them. The shape of parameres is relatively stable in the family, but in some genera, the paramere is elongated, appearing less resistant against lateral deflection. To understand the mechanical limitations of the paramere, we studied its penetration mechanics of the common bed bug, Cimex lectularius. We examined the post-abdominal morphology, paramere geometry and material properties and conducted breaking stress experiments on the paramere under wet and dry conditions. Mechanical property gradients are present with the paramere tip as the stiffest region and the base as the most flexible one. These mechanical properties relate to the presence of Ca, Zn and Si. The basal wing-shaped structure is flexible, enabling it to interlock with the anal region during mating. The paramere is slightly twisted; the tip region is circular in cross-section, and the geometry of the rest is rather complex. In the mechanical tests, wet parameres mainly buckled, while dried parameres broke off. The level of structural failures depended on directions from which the compression forces were applied. Structural, material and mechanical strengthening mechanisms preventing the paramere from mechanical failure are discussed.

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Parasites are integral parts of ecosystem function and important drivers of evolutionary processes. Characterizing ectoparasite diversity is fundamental to studies of host-parasite interactions, evolution, and conservation, and also for understanding emerging disease threats for some vector borne pathogens. With more than 1400 species, bats represent the second most speciose mammalian clade, but their ectoparasite fauna are poorly known for most species. We sequenced mitochondrial Cytochrome Oxidase C subunit I and nuclear 18S ribosomal gene fragments, and used Bayesian phylogenetic analyses to characterize ectoparasite taxon identity and diversity for 17 species of parasitized bats sampled along the Baja California peninsula and in Northwestern Mexico. The sequence data revealed multiple novel lineages of bat bugs (Cimicidae), flies (Nycteribiidae and Streblidae), and ticks (Argasidae). Within families, the new linages showed more than 10% sequence divergence, which is consistent with separation at least at the species level. Both families of bat flies showed host specificity, particularly on Myotis species. We also identified new records for the Baja peninsula of one tick (Carios kelleyi), and of five Streblid bat fly species. One Nycteribiid bat fly haplotype from Pallid bat (Antrozous pallidus) hosts was found throughout the peninsula, suggesting potential long distance co-dispersal with hosts. Different bat bug and tick communities were found in the north and south of the peninsula. This study is the first systematic survey of bat ectoparasites in the Baja California peninsula, revealing novel lineages that are highly genetically differentiated from other parts of North America. For some ectoparasite species, haplotype distributions may reflect patterns of bat migration. This work is a first step in characterizing ectoparasite diversity over the Baja California peninsula, and understanding how ecological and evolutionary interactions shape bat ectoparasite communities among host species in different parts of their ranges.

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An infestation of a Cimicidae (Hemiptera: Cimicidae) member, especially the bed bug, can cause economic loss and impact health. A cost-effective and user-friendly method for identifying the infesting species will help with the early detection and control of infestations. A linear morphometric method is often used, but it requires the examination of many characters and a highly preserved specimen. We conducted a comparative morphometric study of the effectiveness of Cimicidae classification using a single organ, the pronotum, through outline-based and linear morphometric methods. Bat (Stricticimex parvus), human (Cimex hemipterus), and bird (Paracimex sp.) ectoparasites were subject of the study. With both methods, the properties of size and shape were compared and used separately to classify the specimens. Classification analyses of the two methods provided similar results, but more informative variables of size and shape were obtained with the outline-based approach. Size, as analyzed with the outline-based method, could detect sexual dimorphism, and produced better reclassification. The shape variables obtained from the linear measurements were strongly influenced by size variation, much more than the ones obtained from coordinates describing the pronotum contours. Our data suggest that the outline-based approach provides better characterization variables, thus we recommend them for a wider use in other Cimicidae family members.

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Polyctenidae bugs are rarely studied, hematophagous, and highly specialized ectoparasites of bats. There are only 32 described species worldwide, including six species in the Afrotropical region. Knowledge on these parasites is limited, and most studies are restricted to the New World polyctenid species. Here we report additional records of Adroctenes horvathi from Kenya and South Africa, as well as Hypoctenes faini from Rwanda. We present an updated list of published polyctenid records in the Afrotropical region indicating their host specificity and their geographical distribution. We report global infection patterns and sex ratio of polyctenids based on previously published data, including Old and New World species. Lastly, we demonstrate the first molecular phylogeny of Polyctenidae, showing their phylogenetic relationship with the closely related family Cimicidae.

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The bed bug, Cimex lectularius L., is a common ectoparasite found to live among its vertebrate hosts. Antennal segments in bugs are critical for sensing multiple cues in the environment for survival. To determine whether the thermo receptors of bed bugs are located on their antennae; innovative bioassays were created to observe the choice between heated and unheated stimuli and to characterize the response of bugs to a heat source. Additionally, the effect of complete antenectomized segments on heat detection were evaluated. Heat, carbon dioxide, and moisture are cues that are found to activate bed bug behavior; a temperature at 38°C was used to assess the direction/degree at which the insect reacts to the change in distance from said stimulus. Using a lightweight spherical ball suspended by air through a vacuum tube, bed bugs and other insects are able to move in 360° while on a stationary point. Noldus EthoVision XT was used to capture video images and to track the bed bugs during 5-min bioassays. A bioassay was created using four Petri dish arenas to observe bed bug attraction to heat based on antennae segments at 40°C. The purpose of this study was to evaluate the effects of heat on complete antenectomized segments of the antennae. The results in this experiment suggest that bed bugs detect and are attracted to heat modulated by nutritional status. Learning the involvement of antennae segments in heat detection will help identify the location and role of thermoreceptors for bed bug host interaction.

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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.

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Objective To measure the population size and fecundity of the common bed bug Cimex lectularius L. (C. lectularius) after feeding it with the blood obtained from human subjects who have consumed a single dose of ivermectin. Methods  Serial blood samples were obtained from two human subjects at hour 0 (control) and 4-96 hours after they received a single 0.2 mg/kg dose of ivermectin. The blood samples were then fed to 2,273 bed bugs. Bed bug incapacitation rates, fecundity, and population sizes were recorded over a 54-day period. Whole blood ivermectin levels were measured in the human subjects and the insects. Results  The fold change in the size of the control group population over the course of the experiment was found to be 2.16. This was significantly greater (p: <.001) than for all the post-ivermectin feeding groups (range: -11.04-1.43). Two weeks after the experiment, the number of eggs laid per live adult female bed bug per day was 10.74 for controls, which was significantly different (p: <0.001) compared to all the post-ivermectin feeding groups (range: 0-4.28). Conclusions  There were significant reductions in C. lectularius population size and fecundity in insects that fed on blood obtained from human study subjects up to 96 hours after they have consumed a single oral dose of ivermectin.

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BACKGROUND: The common bed bug, Cimex lectularius, is an obligatory blood-feeding ectoparasite that requires a blood meal to molt and produce eggs. Their frequent biting to obtain blood meals and intimate association with humans increase the potential for disease transmission. However, despite more than 100 years of inquiry into bed bugs as potential disease vectors, they still have not been conclusively linked to any pathogen or disease. This ecological niche is extraordinarily rare, given that nearly every other blood-feeding arthropod is associated with some type of human or zoonotic disease. Bed bugs rely on the bacteria Wolbachia as an obligate endosymbiont to biosynthesize B vitamins, since they acquire a nutritionally deficient diet, but it is unknown if Wolbachia confers additional benefits to its bed bug host. In some insects, Wolbachia induces resistance to viruses such as Dengue, Chikungunya, West Nile, Drosophila C and Zika, and primes the insect immune system in other blood-feeding insects. Wolbachia might have evolved a similar role in its mutualistic association with the bed bug. In this study, we evaluated the influence of Wolbachia on virus replication within C. lectularius. METHODS: We used feline calicivirus as a model pathogen. We fed 40 bed bugs from an established line of Wolbachia-cured and a line of Wolbachia-positive C. lectularius a virus-laden blood meal, and quantified the amount of virus over five time intervals post-feeding. The antibiotic rifampicin was used to cure bed bugs of Wolbachia. RESULTS: There was a significant effect of time post-feeding, as the amount of virus declined by ~90% over 10 days in both groups, but no significant difference in virus titer was observed between the Wolbachia-positive and Wolbachia-cured groups. CONCLUSIONS: These findings suggest that other mechanisms are involved in virus suppression within bed bugs, independent of the influence of Wolbachia, and our conclusions underscore the need for future research.

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Due to their high impact on public health, human blood-feeding arthropods are one of the most relevant animal groups. Bacterial symbionts have been long known to play a role in the metabolism, and reproduction of these arthropod vectors. Nowadays, we have a more complete picture of their functions, acknowledging the wide influence of bacterial symbionts on processes ranging from the immune response of the arthropod host to the possible establishment of pathogens and parasites. One or two primary symbiont species have been found to co-evolve along with their host in each taxon (being ticks an exception), leading to various kinds of symbiosis, mostly mutualistic in nature. Moreover, several secondary symbiont species are shared by all arthropod groups. With respect to gut microbiota, several bacterial symbionts genera are hosted in common, indicating that these bacterial groups are prone to invade several hematophagous arthropod species feeding on humans. The main mechanisms underlying bacterium-arthropod symbiosis are discussed, highlighting that even primary symbionts elicit an immune response from the host. Bacterial groups in the gut microbiota play a key role in immune homeostasis, and in some cases symbiont bacteria could be competing directly or indirectly with pathogens and parasites. Finally, the effects climate change, great human migrations, and the increasingly frequent interactions of wild and domestic animal species are analyzed, along with their implications on microbiota alteration and their possible impacts on public health and the control of pathogens and parasites harbored in arthropod vectors of human parasites and pathogens.

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Identification of landscape features that correlate with genetic structure permits understanding of factors that may influence gene flow in a species. Comparing effects of the landscape on a parasite and host provides potential insights into parasite-host ecology. We compared fine-scale spatial genetic structure between big brown bats (Eptesicus fuscus) and their cimicid ectoparasite (Cimex adjunctus; class Insecta) in the lower Great Lakes region of the United States, in an area of about 160,000 km2. We genotyped 142 big brown bat and 55 C. adjunctus samples at eight and seven microsatellite loci, respectively, and inferred effects of various types of land cover on the genetic structure of each species. We found significant associations between several land cover types and genetic distance in both species, although different land cover types were influential in each. Our results suggest that even in a parasite that is almost entirely reliant on its hosts for dispersal, land cover can affect gene flow differently than in the hosts, depending on key ecological aspects of both species.

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BACKGROUND: Residual insecticides interrupt the dispersal of bed bugs (Cimex lectularius, L.), but one of the issues encountered with residual applications is understanding the uptake of active residues by the insect. This study determined permethrin uptake by bed bugs walking on the ActiveGuard® Mattress Liner product, via a combination of video recording in arenas and gas chromatography analyses. RESULTS: The best model for estimating permethrin uptake utilized a covariance model (r2 = 0.469) with two factors: time of exposure (F1,55 = 2.44, P < 0.001) and distance traveled (F1,55 = 0.30, P = 0.0460). Bed bug permethrin uptake was 15.1 (95% CI: 10.3-22.1) ng insect-1 within 1 min exposure, 21.0 (15.0, 31.0) ng insect-1 within 10 min and ≈ 42 (29.8, 60.6) ng insect-1 within ≥50 min exposure. Correcting for percentage recovery, these values would be increased by a factor of 1.21. CONCLUSION: This permethrin-treated fabric provides a surface from which bed bugs begin rapidly to absorb permethrin on contact and within the first 1 cm of travel. Variability in uptake was likely a result of grooming and thigmotaxis, and future work should use quantitative methods to study behaviors and formulations that increase exposure to the toxicant. © 2016 Society of Chemical Industry.

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Few studies have addressed the efficacy of insecticides used against eggs and first-instar nymphs of the bed bug, Cimex lectularius L. (Hemiptera: Cimicidae). Insect eggs are often resistant to insecticides; therefore, information on which products are effective is important. We evaluated the efficacy of four commonly used insecticide sprays applied directly to bed bug eggs. We also evaluated the efficacy of these insecticides to first-instar nymphs exposed to residuals resulting from directly spraying eggs. Temprid SC (beta-cyfluthrin, imidacloprid) was the most effective insecticide at preventing egg hatch (13% hatch rate) for pyrethroid-resistant, field-strain (Jersey City) bed bugs compared with a control (water [99% hatch rate]), Bedlam (MGK-264, sumithrin [84% hatch rate]), Demand CS (lambda-cyhalothrin [91% hatch rate]), and Phantom SC (chlorfenapyr [95% hatch rate]). Demand CS and Temprid SC were most effective at preventing egg hatch (0%) for an insecticide-susceptible (Harold Harlan) strain, followed by Bedlam (28%). Phantom SC produced a hatch rate similar to the control (97% and 96%, respectively). Harold Harlan-strain nymphs showed 100% survival for the control but 0% survival for Bedlam and Phantom SC. Jersey City-strain nymphs showed 100% survival for the control, 99% survival for Bedlam, 0% survival for Demand CS, 4% survival for Phantom SC, and 38% survival for Temprid SC. Demand CS was less effective at preventing hatch (91% hatch rate) of Jersey City-strain nymphs but was the only product to kill all nymphs (0% survival). One of the least effective products for preventing Jersey City-strain egg hatch (Phantom SC, 95% hatch rate) was the second most effective at killing nymphs, leaving only six of 141 alive. These findings indicate that survival of directly sprayed eggs and residually exposed, first-instar nymphs varies by strain, life stage, and product used.

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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.

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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.

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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.

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Bed bugs are hematophagous insects which due to their morphological and biological characteristics are able to easily adapt themselves to human households. The authors describe two cases of dermatitis caused by bed bug bites in the city of Rio de Janeiro, Brazil. Patients presented linear lesions in the usual "breakfast, lunch and dinner" arrangement, suggesting this diagnosis. A visit to their dwellings showed infestation of insects identified as Cimex hemipterus. The knowledge of these insects by the dermatological community will contribute to an accurate diagnosis as well as subsidize the dissemination of information aiming for prevention.

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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.

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