The First FISH-Confirmed Non-Canonical Telomeric Motif in Heteroptera: Cimex lectularius Linnaeus, 1758 and C. hemipterus (Fabricius, 1803) (Hemiptera, Cimicidae) Have a 10 bp Motif (TTAGGGATGG)n.

Fluorescence in situ hybridization (FISH) with two different probes, the canonical insect telomeric sequence (TTAGG)n and the sequence (TTAGGGATGG)n, was performed on meiotic chromosomes of two members of the true bug family Cimicidae (Cimicomorpha), the common bed bug Cimex lectularius Linnaeus, 1758 and the tropical bed bug C. hemipterus (Fabricius, 1803), whose telomeric motifs were not known. In both species, there were no hybridization signals with the first probe, but strong signals at chromosomal ends were observed with the second probe, indicating the presence of a telomeric motif (TTAGGGATGG)n. This study represents the first FISH confirmation of the presence of a non-canonical telomeric motif not only for the infraorder Cimicomorpha but also for the suborder Heteroptera (Hemiptera) as a whole. The present finding is of key significance for unraveling the evolutionary shifts in the telomeric sequences in this suborder.

Protocol for tropical bed bug use as forensic tools.

As blood-feeding insects that feed on human hosts, bed bugs could be used in forensic investigations if they are present at a crime scene with no apparent evidence. This study describes how tropical bed bugs (Cimex hemipterus) can be used as forensic tools to collect valid human DNA samples. Short Tandem Repeat (STR) analysis was performed on collected bed bug samples, whereby the results indicate that the obtained quantities of human DNA are mostly substantial to facilitate a comprehensive genetic profiling process.

Review on the impacts of indoor vector control on domiciliary pests: good intentions challenged by harsh realities.

Arthropod vectored diseases have been a major impediment to societal advancements globally. Strategies to mitigate transmission of these diseases include preventative care (e.g. vaccination), primary treatment and most notably, the suppression of vectors in both indoor and outdoor spaces. The outcomes of indoor vector control (IVC) strategies, such as long-lasting insecticide-treated nets (LLINs) and indoor residual sprays (IRSs), are heavily influenced by individual and community-level perceptions and acceptance. These perceptions, and therefore product acceptance, are largely influenced by the successful suppression of non-target nuisance pests such as bed bugs and cockroaches. Adoption and consistent use of LLINs and IRS is responsible for immense reductions in the prevalence and incidence of malaria. However, recent observations suggest that failed control of indoor pests, leading to product distrust and abandonment, may threaten vector control programme success and further derail already slowed progress towards malaria elimination. We review the evidence of the relationship between IVC and nuisance pests and discuss the dearth of research on this relationship. We make the case that the ancillary control of indoor nuisance and public health pests needs to be considered in the development and implementation of new technologies for malaria elimination.

Bed bug control with various dusts: Efficacy comparison between silicon dioxide, diatomaceous earth, and Sommières earth.

Bed bugs are considered a major public health problem in industrialized countries. Usually, bed bug infestations are managed using a combination of physical and chemical methods. In recent years, new strategies for bed bug control have emerged, particularly the use of dusts like diatomaceous earth and silicon dioxide. However, in Europe, the use of silicon dioxide is restricted to professional, while diatomaceous earth can be harmful to the lungs. This study aimed to assess bed bug mortality rates associated with Sommières earth, green clay, talc, and sodium bicarbonate compared to silicon dioxide and diatomaceous earth from a pest management company, diatomaceous earth for litter conditioner, and diatomaceous earth from a supermarket. We tested permanent exposure, short exposure, horizontal transfer and repellent effect on two bed bug colonies. Sommières earth demonstrated efficacy ranging from 75% to 100% in permanent and short exposures, similar to the efficacy of diatomaceous earth from the pest management company. On the contrary, diatomaceous earth for litter conditioner and diatomaceous earth from a supermarket, green clay, talc, and sodium bicarbonate were found to be ineffective. This study demonstrates, for the first time, the efficacy of Sommières earth against bed bugs, but also highlights the variability in efficacy of diatomaceous earths on bed bugs depending on their quality.

Title: Lutte contre les punaises de lit avec des poudres : comparaison de l'efficacité du dioxyde de silicium, de la terre de diatomée et de la terre de Sommières. Abstract: Les punaises de lit sont considérées comme un problème de santé publique majeur dans les pays industrialisés. Habituellement, les infestations de punaises de lit sont gérées en utilisant une combinaison de méthodes physiques et chimiques. Ces dernières années, de nouvelles stratégies de lutte contre les punaises de lit ont vu le jour, notamment l'utilisation de poudres comme la terre de diatomées et le dioxyde de silicium. Cependant, en Europe, l'usage du dioxyde de silicium est réservé aux professionnels tandis que la terre de diatomées peut être nocive pour les poumons. Cette étude visait à évaluer les taux de mortalité des punaises de lit associés à la terre de Sommières, à l'argile verte, au talc et au bicarbonate de sodium par rapport au dioxyde de silicium, à la terre de diatomées d'une entreprise de lutte antiparasitaire, à la terre de diatomées pour conditionneur de litière et à la terre de diatomées d'un supermarché. Nous avons testé l'exposition permanente, l'exposition courte, le transfert horizontal et l'effet répulsif sur deux colonies de punaises de lit. La terre de Sommières a démontré une efficacité allant de 75% à 100% en exposition permanente et courte, similaire à l'efficacité de la terre de diatomées d'une entreprise de lutte antiparasitaire. Au contraire, la terre de diatomées pour conditionneur de litière et la terre de diatomées d'un supermarché, l'argile verte, le talc et le bicarbonate de sodium se sont révélés inefficaces. Cette étude démontre, pour la première fois, l'efficacité de la terre de Sommières contre les punaises de lit mais met également en évidence la variabilité de l'efficacité des terres de diatomées sur les punaises de lit en fonction de leur qualité.

Morphometric Characteristics and Species Identification of Bed Bugs (Hemiptera: Cimicidae) in Eastern Iran.

Background: The common bed bugs, Cimex lectularius, and tropical bed bugs, Cimex hemipterus are the primary species of public health importance in the family Cimicidae. This study aimed to determine the morphometric criteria and prevalent species of bed bugs in eastern Iran. Methods: Bed bugs were collected from March 2021 to June 2022 from infested residential buildings and apartments in eastern Iran, including North Khorasan, Khorasan Razavi, and Sistan and Baluchistan Provinces. The morphological and morphometrical characteristics were used to identify collected bed bugs at inter- and intra-specific levels. Results: A total of 34 isolates comprising 127 adult bed bugs were collected from Bojnord, Mashhad, Neishabur, Taibad, Sabzevar, Kashmer, Zahedan, Saravan, Rask, Pishin and Chabahar. Of these, 33 isolates (n=124) were found to be tropical bed bugs, C. hemipterus, and one population (n=3) was identified as C. lectularius. The index pronotal width/length ratio was calculated from 2.72 to 2.94 and 1.98 to 2.47 for C. lectularius and C. hemipterus, respectively. The length/width ratio of the hind femur was 3.365 in C. hemipterus and 4.267 in C. lectularius. The ratio of length/width of the third femur (F3 l/w) between populations of C. hemipterus was different, and this difference was statistically significant (P< 0.05). Conclusion: The results of this study indicated that C. hemipterus was the dominant bed bug species in the east of Iran and provided more morphometric criteria of C. hemipterus for researchers to identify the species and determine the intraspecific variations in the present and future.

Sexual conflict and sexual networks in bed bugs: the fitness cost of traumatic insemination, female avoidance and male mate choice.

Sexual conflict is prevalent among animals and is primarily caused by the fact that the optimal mating rates are often higher in males than in females. While there is a growing appreciation that females can also gain from multiple matings, we still know relatively little about which sex controls the observed mating rates and how close it is to the optimal female mating rates. To address this issue, we tracked female bed bugs (Cimex lectularius) inseminated daily versus weekly and found that weekly inseminated females lived longer and produced over 50% more offspring. In a follow-up experiment employing a social network framework, we placed 24 bed bugs into a semi-naturalistic arena and recorded all sexual interactions. While recently inseminated females did not avoid males more often, they were more frequently rejected by males. Finally, we tracked avoidance behaviour in a single cohort of female bed bugs as they received six successive daily inseminations. Avoidance rates increased and insemination durations decreased with increasing number of prior inseminations. Overall, our results indicate high costs of polyandry. Although females possess some plastic avoidance strategies, the observed rates of insemination fall closer to the male rather than female optimum.

A novel prolixicin identified in common bed bugs with activity against both bacteria and parasites.

The hematophagous common bed bug, Cimex lectularius, is not known to transmit human pathogens outside laboratory settings, having evolved various immune defense mechanisms including the expression of antimicrobial peptides (AMPs). We unveil three novel prolixicin AMPs in bed bugs, exhibiting strong homology to the prolixicin of kissing bugs, Rhodnius prolixus, and to diptericin/attacin AMPs. We demonstrate for the first time sex-specific and immune mode-specific upregulation of these prolixicins in immune organs, the midgut and rest of body, following injection and ingestion of Gr+ (Bacillus subtilis) and Gr- (Escherichia coli) bacteria. Synthetic CL-prolixicin2 significantly inhibited growth of E. coli strains and killed or impeded Trypanosoma cruzi, the Chagas disease agent. Our findings suggest that prolixicins are regulated by both IMD and Toll immune pathways, supporting cross-talk and blurred functional differentiation between major immune pathways. The efficacy of CL-prolixicin2 against T. cruzi underscores the potential of AMPs in Chagas disease management.

Sexual conflict and social networks in bed bugs: effects of social experience.

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.

Activation of immune pathways in common bed bugs, Cimex lectularius, in response to bacterial immune challenges - a transcriptomics analysis.

The common bed bug, Cimex lectularius, is an urban pest of global health significance, severely affecting the physical and mental health of humans. In contrast to most other blood-feeding arthropods, bed bugs are not major vectors of pathogens, but the underlying mechanisms for this phenomenon are largely unexplored. Here, we present the first transcriptomics study of bed bugs in response to immune challenges. To study transcriptional variations in bed bugs following ingestion of bacteria, we extracted and processed mRNA from body tissues of adult male bed bugs after ingestion of sterile blood or blood containing the Gram-positive (Gr+) bacterium Bacillus subtilis or the Gram-negative (Gr-) bacterium Escherichia coli. We analyzed mRNA from the bed bugs' midgut (the primary tissue involved in blood ingestion) and from the rest of their bodies (RoB; body minus head and midgut tissues). We show that the midgut exhibits a stronger immune response to ingestion of bacteria than the RoB, as indicated by the expression of genes encoding antimicrobial peptides (AMPs). Both the Toll and Imd signaling pathways, associated with immune responses, were highly activated by the ingestion of bacteria. Bacterial infection in bed bugs further provides evidence for metabolic reconfiguration and resource allocation in the bed bugs' midgut and RoB to promote production of AMPs. Our data suggest that infection with particular pathogens in bed bugs may be associated with altered metabolic pathways within the midgut and RoB that favors immune responses. We further show that multiple established cellular immune responses are preserved and are activated by the presence of specific pathogens. Our study provides a greater understanding of nuances in the immune responses of bed bugs towards pathogens that ultimately might contribute to novel bed bug control tactics.

Optimization and establishment of laboratory rearing conditions for Cimex lectularius L. against variable temperature and relative humidity.

Emerging infestations of bed bugs are affecting normal human lifestyle globally. This study has been designed to optimize the rearing conditions for Cimex lectularius L. (Hemiptera), to support the scientific research on them. Bed bugs have been projected onto three different temperature (20 °C, 25 °C, and 30 °C) and relative humidity (50%, 70%, and 90%) conditions to check their overall growth and survival rate. Adult mortality, weight loss, egg laying, percentage hatching, hatching initiation and completion, nymph mortality, and molting have been evaluated to optimize the best conditions. The temperature at 25 °C with 90% RH showed minimum mortality for adults (female 13.33 ± 3.33% and male 6.67 ± 3.33%) and nymphs (13.33 ± 3.33%), while maximum egg laying (40.33 ± 1.86), with highest percentage hatching (98.23 ± 0.58%). At 30 °C with 90% RH, hatching initiation and completion (5.19 ± 0.12 days and 7.23 ± 0.16 days) as well as molting initiation and completion (3.73 ± 0.12 days and 7.00 ± 0.24 days) were found to be fastest. Thus, it can be concluded that 25 °C with 90% RH is ideal for rearing of adults and 30 °C with 90% RH is appropriate for rapid growth of nymphs.

First official report of bed bug (Hemiptera, Cimicidae) infestations in Algeria.

Background: Bed bugs are hematophagous insects with a long history of presence in human communities. Over the last three decades, infestations by bed bugs in human dwellings have drastically increased, leading to a rise in bed bug concerns. Nevertheless, very little is known about the bed bug species and their population diversity in Algeria. Method: A pilot entomological inventory was performed in May 2019 in Tizi Ouzou, in northern Algeria. The gathered bed bug specimens were identified by morphological and molecular approaches, followed by neighbor-joining and network phylogenetic analyses. Results: A total of seven out of 12 requested locations were allowed to inspect for bed bug infestation. Of these, three locations were found with active bed bug infestations. A total of 145 specimens belonging to different life stages [egg (21), nymph (74), adult male (17), and female (33)] were collected and analyzed using morphological and molecular approaches. The adult specimens were identified as Cimex lectularius according to specific morphological criteria, most importantly the pronotum laterally expanded with more flattened extreme margins. Morphological identification of the adults was confirmed further by conventional PCR targeting 450 bp fragment of the COI gene. All the nymphs and eggs were also molecularly identified as C. lectularius. Neighbor-Joining phylogenetic tree reconstructed with the collected specimens provides clues on the presence of two closely phylogenetic groups. The first one gathers our samples of Algeria with previously reported COI haplotype sequences from Asian, European, and North American countries. The second group encompasses a lesser-documented haplotype reported in Europe and Central America. These findings were further confirmed by network analysis. Conclusions: These results provide evidence of established C. lectularius infestation in Algeria and its potential dispersal capacity by travelers or immigrants and will help future management of these ectoparasites.

Morphological, molecular, and MALDI-TOF MS identification of bed bugs and associated Wolbachia species from Cameroon.

After vanishing from the public eye for more than 50 years, bed bugs have resurged to become one of the most widely discussed and heavily researched insect pests in the world. This study presents the basic information of infestations of tropical bed bugs, Cimex hemipterus (Hemiptera: Cimicidae), in Cameroon. A total of 248 immature stage and adult bed bug specimens were collected from households and a travel agency in Yaoundé and Douala, Cameroon. The ability of MALDI-TOF MS to identify bed bugs was tested using heads for adults and cephalothoraxes for immature stages. Microorganism screening was performed by qPCR and confirmed by regular PCR and sequencing. Based on morphometrical criteria, four stages of immature bed bugs are represented. Of the 248 bed bug specimens morphologically identified as Cimex hemipterus, 246 (77 males, 65 females and 104 immature specimens) were submitted to MALDI-TOF MS analysis. Of the 222 adults and immature specimens tested, 122 (59.9 %) produced good quality MALDI-TOF MS spectra (35 adults and 87 immature specimens). Blind testing allowed species level identification of 98.21 % of adult and immature C. hemipterus. Among the bacteria tested, only Wolbachia DNA was found in 12/246 (4.8 %) bed bugs. More surveys in the country are warranted to assess the true level of bed bug infestations, in order to take appropriate action for their control.

Pharmacokinetics of fluralaner as a systemic drug to control infestations of the common bed bug, Cimex lectularius, in poultry facilities.

BACKGROUND: Bed bug infestations are re-emerging in the poultry industry throughout the USA. Although the impacts of bed bugs on birds' health and welfare are poorly understood, adverse outcomes are expected, including stress, anemia, infections and lower production rates. Worker welfare is also an important consideration in commercial poultry farms. A limited number of insecticides are available for use in the complex spatial environment of commercial farms. Systemic drugs have the potential to overcome the limitations of existing pest management tactics. A recent study showed that fluralaner administered to chickens caused high levels of mortality in bed bugs. METHODS: To further understand the efficacy of this approach, we evaluated the pharmacokinetics of an oral solid formulation of fluralaner in 11 chickens and quantified its plasma concentration in chickens using UPLC/MS. We administered fluralaner to chickens with two doses of Bravecto® (each 0.5 mg/kg body mass) via gavage 1 week apart and evaluated its efficacy on bed bugs that fed on medicated chickens for up to 28 days post-treatment. RESULTS: Bed bugs that fed on fluralaner-treated chickens experienced > 50% mortality within 30 min of the administration of Bravecto and 100% mortality 2 days post-treatment. Mortality slowly declined to 66.6% by day 28. Fluralaner was quantifiable in the hens' plasma for at least 28 days post-treatment. The treatment resulted in maximal plasma concentrations (Cmax) of 106.4 ng/ml around day 9.0 (Tmax), substantially higher than the LC90, the concentration needed to kill 90% of the bed bugs. CONCLUSIONS: Fluralaner appears to be a promising candidate for bed bug control in poultry farms, with a treatment effect lasting at least 28 days.

Bed Bugs (Hemiptera, Cimicidae): A Global Challenge for Public Health and Control Management.

Bed bugs, Cimex lectularius, and C. hemipterus are among the most common ectoparasites in human life worldwide. They feed on humans of all ages and sexes across all socioeconomic levels. Bed bugs' blood feeding is responsible for a wide range of clinical manifestations varying from minor reactions to bullous eruptions or severe allergies. In addition, they are responsible for considerable psychological distress. Therefore, diagnosis of bed bug bites and their consequence manifestations is beneficial in adapting remedies and treatment protocols advised by clinicians. So far, there is regrettably no definitive way to control these ectoparasites despite extensive efforts of public health authorities to manage them. An overview of the literature and medical documents gathered from bed bug-infested patients referred to the Parasitology and Dermatology departments of Avicenne Hospital (Bobigny, France) allowed us to document and illustrate a range of clinical disorders and psychological concerns caused by bed bugs' bites and their clinical diagnosis. We also review the available tools currently used to control the bed bugs and present potential candidate methods for their successful eradication.

Cimicids of Medical and Veterinary Importance.

Members of the Cimicidae family are significant pests for mammals and birds, and they have attracted medical and veterinary interest. A number of recent studies have investigated bed bugs, due to their dramatic resurgence all over the world. Indeed, bed bugs are of significant public health and socioeconomic importance since they lead to financial burdens and dermatological complications and may have mental and psychological consequences. It is important to note that certain cimicids with a preference for specific hosts (birds and bats) use humans as an alternative host, and some cimicids have been reported to willingly feed on human blood. In addition, members of the Cimicidae family can lead to economic burdens and certain species are the vectors for pathogens responsible for diseases. Therefore, in this review, we aim to provide an update on the species within the Cimicidae family that have varying medical and veterinary impacts, including their distribution and their associated microorganisms. Various microbes have been documented in bed bugs and certain important pathogens have been experimentally documented to be passively transmitted by bed bugs, although no conclusive evidence has yet associated them with epidemiological outbreaks. Additionally, among the studied cimicids (bat bugs, chicken bugs, and swallow bugs), only the American swallow bug has been considered to be a vector of several arboviruses, although there is no proven evidence of transmission to humans or animals. Further studies are needed to elucidate the reason that certain species in the Cimicidae family cannot be biologically involved in transmission to humans or animals. Additional investigations are also required to better understand the roles of Cimicidae family members in the transmission of human pathogens in the field.

Volatile Organic Compounds: A Promising Tool for Bed Bug Detection.

The recent decades' resurgence of bed bugs as a public health concern in industrialized countries has driven an increased interest on new sustainable insecticide-free methods to monitor and control these ectoparasites. Current methods of detection rely mainly on visual inspection or canine scent detection, which are methods that are time-consuming, require experience, are non-specific or require costly mission repetitions. Volatile organic compounds (VOCs) are considered an environmentally friendly alternative and a promising approach for bed bug detection. An overview of the released literature on VOCs, their chemical characteristics and their role in bed bugs' intra- and inter-species communications allowed us to highlight the identification of 49 VOCs in Cimex lectularius (23 molecules) and C. hemipterus (26), which are emitted by both sexes during diverse compartments including aggregation (46), mating (11), defense (4), etc., and all life stages including exuviae or dead bed bugs as a principal indicator of infestation. The latter has a great importance for application of these semiochemicals in successful detection and control management of bed bugs and to prevent their further dispersion. This approach has the advantage of more reliability compared to conventional detection methods with no need for repeated inspections, household furniture moving or resident rehousing for bed bugs' VOC detection, which are commonly performed by active or passive sampling with absorbing tubes and analyzed by gas chromatography-based analytical platforms.

Detection of Arthropod-Borne Bacteria and Assessment of MALDI-TOF MS for the Identification of Field-Collected Immature Bed Bugs from Mauritania.

Human infestations by bed bugs have upsurged globally in recent decades, including in African countries, where recent reports pointed out an increase in infestation. Sympatric dwelling has been described for two species of bed bug parasitizing humans: Cimex hemipterus (the tropical bed bug) and C. lectularius. Identification of these two species is based on morphological characteristics, and gene sequencing, and may also rely on Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF MS). The present work aimed to assess whether MALDI-TOF MS was applicable for species level identification of immature stages of Cimex. Arthropods were collected in domestic settings in Nouakchott, Mauritania. Identification used morphological keys and MALDI-TOF MS identification was assessed for immature stages. Quantitative PCR and sequencing assays were used to detect arthropod-associated bacteria in each specimen. A total of 92 arthropods were collected, all morphologically identified as C. hemipterus (32 males, 14 females and 45 immature stages). A total of 35/45 specimens produced good quality MALDI-TOF MS spectra. Analysis allowed species level identification of all immature C. hemipterus after their spectra were entered into our in-house MALDI-TOF MS arthropod spectra database. Molecular screening allowed detection of Wolbachia DNA in each specimen. These results suggested that MALDI-TOF MS is a reliable tool for species level identification of Cimex specimens, including immature specimens. Future studies should assess this approach on larger panels of immature specimens for different Cimex species and focus on the precise staging of their different immature developmental stages.

Characterization of New Defensin Antimicrobial Peptides and Their Expression in Bed Bugs in Response to Bacterial Ingestion and Injection.

Common bed bugs, Cimex lectularius, can carry, but do not transmit, pathogens to the vertebrate hosts on which they feed. Some components of the innate immune system of bed bugs, such as antimicrobial peptides (AMPs), eliminate the pathogens. Here, we determined the molecular characteristics, structural properties, and phylogenetic relatedness of two new defensins (CL-defensin1 (XP_024085718.1), CL-defensin2 (XP_014240919.1)), and two new defensin isoforms (CL-defensin3a (XP_014240918.1), CL-defensin3b (XP_024083729.1)). The complete amino acid sequences of CL-defensin1, CL-defensin2, CL-defensin3a, and CL-defensin3b are strongly conserved, with only minor differences in their signal and pro-peptide regions. We used a combination of comparative transcriptomics and real-time quantitative PCR to evaluate the expression of these defensins in the midguts and the rest of the body of insects that had been injected with bacteria or had ingested blood containing the Gram-positive (Gr+) bacterium Bacillus subtilis and the Gram-negative (Gr-) bacterium Escherichia coli. We demonstrate, for the first time, sex-specific and immunization mode-specific upregulation of bed bug defensins in response to injection or ingestion of Gr+ or Gr- bacteria. Understanding the components, such as these defensins, of the bed bugs' innate immune systems in response to pathogens may help unravel why bed bugs do not transmit pathogens to vertebrates.

Comparative Efficacy of a Fungal Entomopathogen with a Broad Host Range against Two Human-Associated Pests.

The ability of a fungal entomopathogen to infect an insect depends on a variety of factors, including strain, host, and environmental conditions. Similarly, an insect's ability to prevent fungal infection is dependent on its biology, environment, and evolutionary history. Synanthropic pests have adapted to thrive in the indoor environment, yet they arose from divergent evolutionary lineages and occupy different feeding guilds. The hematophagous bed bug (Cimex lectularius) and omnivorous German cockroach (Blattella germanica) are highly successful indoors, but have evolved different physiological and behavioral adaptations to cope with the human-built environment, some of which also reduce the efficacy of fungal biopesticides. In order to gain greater insight into the host barriers that prevent or constrain fungal infection in bed bugs and German cockroaches, we tested different doses of Beauveria bassiana GHA through surface contact, topical application, feeding, and injection. Bed bugs were generally more susceptible to infection by B. bassiana with the mode of delivery having a significant impact on infectivity. The German cockroach was highly resilient to infection, requiring high doses of fungal conidia (>8.8 × 104) delivered by injection into the hemocoel to cause mortality. Mortality occurred much faster in both insect species after exposure to surfaces dusted with dry conidia than surfaces treated with conidia suspended in water or oil. These findings highlight the importance of developing innovative delivery techniques to enhance fungal entomopathogens against bed bugs and cockroaches.