Tropical Bed Bug Research Articles

Development of molecular diagnostic protocols for simultaneous identification of common bed bugs (Cimex lectularius) and tropical bed bugs (Cimex hemipterus)

BackgroundThe resurgence of two bed bug species, the common bed bug (Cimex lectularius Linnaeus, 1758) and tropical bed bug (Cimex hemipterus Fabricius, 1803), in the same geographical regions has been frequently reported recently. Consequently, the rapid identification of these species is crucial for implementing targeted capture traps and tailored pyrethroid resistance diagnosis, due to differences in genetic and physiological traits.MethodsTo develop molecular diagnostic methods, distinct protocols were established for multiplex PCR and loop-mediated isothermal amplification (LAMP) using species-specific primers based on species-specific segments of internal transcribed spacer 2 sequences. These methods were optimized for rapid and accurate identification of the two bed bug species.ResultsBoth multiplex PCR and LAMP protocols were effective in simultaneously identifying the two bed bug species, even when utilizing DNA released from dead specimens. Notably, the straightforward procedure and minimal time commitment of LAMP suggest its potential for rapid and accurate diagnosis of bed bugs in the field. The diagnostic accuracy of these methods was validated through a blind test.ConclusionsThe multiplex PCR and LAMP protocols lay the foundation for rapid and accurate field identification of bed bug species, enabling the use of appropriate traps and the detection of species-specific pyrethroid resistance mutations. This approach ensures effective management tailored to the unique characteristics of each bed bug species.Graphical

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.

Efficacy of plant-based products and nonconventional pesticides for the management of tropical bed bug.

Insecticide resistance is widespread in global bed bug populations. Both common bed bugs and tropical bed bugs are pyrethroid-resistant among most field populations. Plant-based products and nonconventional pesticides offer minimal-risk strategies for managing bed bug resistance, but this strategy has yet to be formally evaluated in Cimex hemipterus (F.) (Hemiptera: Cimicidae). Here, several commercial plant-based formulations (Cedarcide, EcoRaider (also known as EcoVenger), EcoSMART, and Bio-D), a novel product, Provecta, and a pyrethroid insecticide, Pesguard FG161 were tested against pyrethroid-susceptible and resistant strains of C. hemipterus using direct spray, residual exposure, and egg dipping assays. Direct spray treatments outperform residual applications against all tested bed bug strains. Cedarcide exhibits the highest consistency in eliminating bed bugs, followed by EcoRaider, EcoSMART, and Provecta that outperform Bio-D and Pesguard FG161. In comparison to Pesguard FG161, all plant-based insecticide products and Provecta showed higher efficacy against pyrethroid-resistant strains. Although effective, product efficacy varies in terms of speed. Cedarcide kills all bed bugs within 1 min after spraying; however, other products can take up to 9 days to achieve 100% mortality. The efficacy of all products was reduced when evaluated on fabric surface (42%-65% mortality). Cedarcide and EcoRaider reduced egg hatchability by 37%-73% and 47%-70%, respectively. This study suggests that certain plant-based insecticides and an unconventional insecticide can serve as alternative direct spray treatments for managing tropical bed bugs, though their residual effects are limited.

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.

Efficacy of steam and diatomaceous earth dust against the tropical bed bug Cimex hemipterus (F.) under laboratory and field conditions.

Over the past two decades, bed bugs (Cimex spp.) have resurged as common urban pests around the world. The search for efficient and safe control measures has become a key interest among researchers, manufacturers, and pest control professionals. In this study, we evaluated and compared the efficacy of steam, diatomaceous earth (DE) dust, and a combination of both against tropical bed bugs (Cimex hemipterus (F.)) under laboratory and field conditions. In the laboratory study, the mortality of bed bugs after 2 days of exposure to DE dust was 100%. When bed bugs stayed on the surface of an object or in cracks, a brief steam treatment (1 s) caused 100% mortality. However, when bed bugs were hidden under a fabric cover, steam application for 10 s only caused 89 ± 6% mortality. Bed bugs that survived steam treatment exhibited reduced feeding activity. In a 14-week long study, there was no significant difference in the reduction rate of bed bugs between steam treatment and DE dust treatment. A 37-week long control study showed that steam and steam plus DE dust treatments eliminated 97-100% of the infestations. Applying steam and DE dust are effective strategies for eliminating natural tropical bed bug infestations. Continuous follow-up monitoring and treatment until no bed bugs are found are crucial in completely eliminating the infestation of tropical bed bugs. © 2024 Society of Chemical Industry.

Genome of tropical bed bug Cimex hemipterus (Cimicidae, Hemiptera) reveals tetraspanin expanded in bed bug ancestor.

Cimex species are ectoparasites that exclusively feed on warm-blooded animals such as birds and mammals. Three cimicid species are known to be persistent pests for humans, including the tropical bed bug Cimex hemipterus, common bed bug Cimex lectularius, and Eastern bat bug Leptocimex boueti. To date, genomic information is restricted to the common bed bug C. lectularius, which limits understanding their biology and to provide controls of bed bug infestations. Here, a chromosomal-level genome assembly of C. hemipterus (495 Mb [megabase pairs]) contained on 16 pseudochromosomes (scaffold N50 = 34 Mb), together with 9 messenger RNA and small RNA transcriptomes were obtained. In comparison between hemipteran genomes, we found that the tetraspanin superfamily was expanded in the Cimex ancestor. This study provides the first genome assembly for the tropical bed bug C. hemipterus, and offers an unprecedented opportunity to address questions relating to bed bug infestations, as well as genomic evolution to hemipterans more widely.

Species identification and pyrethroid resistance genotyping of recently resurgent Cimex lectularius and Cimex hemipterus in Korea.

The global resurgence of bed bug infestations, exacerbated by increasing international travel, trade, and insecticide resistance, has significantly impacted Korea. This study identified the bed bug species and performed pyrethroid resistance genotyping of recently resurgent bed bugs in Korea. Thirty-one regional bed bug samples were collected from 5 administrative regions: Gyeonggi-do (n=14), Seoul (n=13), Busan (n=2), Jeonllanam-do (n=1), and Chungcheongbuk-do (n=1). The samples underwent morphological and molecular identification. Twenty-four regional samples (77.4%) were identified as the tropical bed bug, Cimex hemipterus, and the remaining 7 regional samples (22.6%) were identified as the common bed bug, Cimex lectularius. The C. hemipterus regional samples carried at least three mutations associated with knockdown resistance (kdr), including 2 super-kdr mutations. The 7 C. lectularius regional samples possessed at least one of the 3 kdr-related mutations associated with pyrethroid resistance. This study confirms that the prevalent bed bug species recently in Korea is C. hemipterus, replacing the previously endemic C. lectularius. Additionally, the rise in bed bug populations with pyrethroid resistance underscores the necessity of introducing alternative insecticides.

Population genetic structure of tropical bed bug (Hemiptera: Cimicidae) populations and their breeding pattern in Iraq.

A study was conducted to investigate the population genetic structure and breeding pattern of 140 tropical bed bugs, Cimex hemipterus (F.) (Hemiptera: Cimicidae), collected from 14 infested sites in major cities in Iraq. The samples were genotyped using a set of 7 polymorphic microsatellite markers. High genetic variety was seen among populations, with an average of 2-9 alleles per locus. The number of alleles across 7 microsatellite loci was between 6 and 18. There was a notable disparity in the alleles per loci when comparing the overall population to those within it. The overall population exhibited an average observed heterozygosity of 0.175 and an average expected heterozygosity of 0.730. Among the population, the average observed heterozygosity was 0.173, while the average expected heterozygosity was 0.673. Analysis of molecular variance (AMOVA) revealed that 93% of the genetic variability was within the populations, and 7% was among them. The genetic differentiation coefficient (FST = 0.045), indicates a low degree of genetic differentiation and a high degree of inbreeding (FIS = 0.761), as indicated by notably significant positive inbreeding coefficients. Admixed individuals were revealed using STRUCTURE and neighbor-joining phylogenetic trees, demonstrating moderate gene flow between populations and a lack of genetic structure in the regional groups. Thus, both active dispersion and human-mediated dispersion possess the potential to influence the low population genetic structure of tropical bed bug C. hemipterus populations in Iraq, which can have implications toward tropical bed bug and management strategies.

Repellent effect of two anthranilates against the tropical bed bug Cimex hemipterus (Hemiptera: Cimicidae) demonstrated in laboratory assays.

The bites and blood sucking of bed bugs (Cimex spp.) (Hemiptera: Cimicidae) pose a serious threat to human physical and mental health. Application of an effective repellent can prevent or reduce bed bug bites. Previous studies on repellent screening mainly focused on Cimex lectularius L. In this study, we investigated the repellent effect of two safe food additives, ethyl anthranilate (EA) and butyl anthranilate (BA), against Cimex hemipterus (F.), and also explored the role of antennae and mouthparts on C. hemipterus perception of repellents. Both EA and BA had a strong repellent effect against tropical bed bugs and their repellency was similar or lower than that of N,N-diethyl-3-methyl benzoyl amide, depending on whether or not a CO2 source was present. EA had higher repellency than BA and exhibited repellency to C. hemipterus for 3 days when a CO2 source was present. C. hemipterus avoided resting on 20% EA- and BA-treated harborages. Applying 20% EA and BA on rabbit skin significantly reduced the blood intake of C. hemipterus within 2 h. C. hemipterus could perceive EA and BA after their antennae or mouthparts or both antennae and mouthparts were removed. Both EA and BA had strong repellency against C. hemipterus, with EA being more repellent. Ablation of antennae and mouthparts did not affect the perception of EA and BA by C. hemipterus. © 2023 Society of Chemical Industry.

Human profiling from STR and SNP analysis of tropical bed bug, Cimex hemipterus, for forensic science

Tropical bed bugs, Cimex hemipterus, which commonly feeds on human blood, may be useful in forensic applications. However, unlike the common bed bug, Cimex lectularius, there is no information regarding tropical bed bug, C. hemipterus, being studied for its applications in forensics. Thus, in this study, lab-reared post-feeding tropical bed bugs were subjected to Short Tandem Repeat (STR) and Single Nucleotide Polymorphism (SNP) analyses to establish the usage of tropical bed bugs in forensics. Several post-feeding times (0, 5, 14, 30, and 45 days) were tested to determine when a complete human DNA profile could still be obtained after the bugs had taken the blood meal. The results showed that complete STR and SNP profiles could only be obtained from the D0 sample. The profile completeness decreased over time, and partial STR and SNP profiles could be obtained up to 45 days post-blood meal. The generated SNP profiles, complete or partial, were also viable for HIrisPlex-S phenotype prediction. In addition, field-collected bed bugs were also used to examine the viability of the tested STR markers, and the STR markers detected mixed profiles. The findings of this study established that the post-blood meal of tropical bed bugs is a suitable source of human DNA for forensic STR and SNP profiling. Human DNA recovered from bed bugs can be used to identify spatial and temporal relations of events.

Historical and Contemporary Control Options Against Bed Bugs, Cimex spp.

Bed bugs (Hemiptera: Cimicidae) are an important group of obligate hematophagous urban insect pests. The global resurgence of bed bugs, involving the common bed bug, Cimex lectularius L., and the tropical bed bug, Cimex hemipterus (F.), over the past two decades is believed to be primarily due to the development of insecticide resistance, along with global travel and poor pest management, which have contributed to their spread. This review examines and synthesizes the literature on bed bug origins and their global spread and the literature on historical and contemporary control options. This includes bed bug prevention, detection and monitoring, nonchemical and chemical control methodologies (and their limitations), and potential future control options. Future research needs are highlighted, especially the factors behind the modern resurgence, the necessity of identifying differences between the two bed bug species relevant to control, and the need to improve insecticide test protocols and management strategies.

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.

Susceptibility and fecundity of Bedbugs (Cimex hemipterus) from Yaba College of Technology Lagos exposed to Selected Classes of Insecticides: A short report

Background: Tropical bed bug, Cimex hemipterus, is a nocturnal obligate blood-sucking ectoparasite of humans that is highly prevalent in Nigeria. Bed bug controls heavily rely on the application of several insecticide-based formulations. Insecticide resistance in bed bug populations has been widely reported. In this study, we assessed the mortality of bed bugs following exposure to different classes of insecticides. Methodology: Active adult female bed bugs were collected from students’ halls of residence and exposed to DDT (4%), permethrin (0.75), bendiocarb (0.1) and malathion (4%) insecticide-impregnated papers. Mortality and number of eggs laid by each exposed group were recorded at 90 minutes, 24hrs, 48hrs and 72hrs. Results: The impact of the different assays on bugs mortality revealed DDT to produce a time-dependent outcome with the highest mortality rate [(36.7% (Cl 32.7±38.9)], this was followed by Bendiocarb [31.6% (CI 27.2±35.1)]. In comparison, permethrin and malathion produced the same effect [26.7% (CI 23±30.5)] 24 hours post-exposure. In contrast, malathion and bendiocarb impacted the most mortality [53.3% (49.3±58.7) and 46.6%(42.2±49.5)], followed by permethrin [18.9% (CI 44.6±56.8) after 72 hours. The relative numbers of eggs produced by bedbugs in the DDT, bendiocarb and malathion assays were lower than what was observed in the control and permethrin group 24 hours after exposure. The highest reduction in egg production was observed in the malathion exposed group 48- and 72-hours post-exposure compared to the other insecticides and the control group. Conclusion: This study revealed suspected insecticide resistance to all classes of insecticide used on bed bug populations in Lagos State, Nigeria.

Identification and Validation of Reference Genes for Expression Analysis Using qRT-PCR in Cimex hemipterus (Hemiptera: Cimicidae).

The tropical bed bug, Cimex hemipterus (F.) (Hemiptera: Cimicidae) is an important public-health pest that feeds on the blood of humans and some other animals. To explore the function of the target genes of C. hemipterus, it is essential to select suitable reference genes for the accurate quantification of gene expression. Here, we selected 10 frequently used reference genes in insects and evaluated their stability in C. hemipterus under various biotic (developmental stage, sex, and tissue) and abiotic (gas stimulation and temperature) conditions through RefFinder (which integrates four computational programs: geNorm, NormFinder, BestKeeper, and ∆Ct). Results indicate that the optimal combination of reference genes for each experimental condition was as follows: RPL8 and EF1α for the developmental stage (eggs, early instar nymphs, late instar nymphs, and adults), RPL8 and RPS16 for adult sex, RPL8 and RPL11 for adult tissue (head, thorax, abdomen, and legs), RPL8 and β-tubulin for gas stimulation (air and carbon dioxide), and RPL8 and NADH for temperature (0, 5, 17, 30, and 37 °C). Finally, the expression pattern of the HSP70 and GR21 genes were analyzed, and the results highlight the importance of appropriate reference-gene selection. Our results provide a comprehensive list of optimal reference genes from C. hemipterus for the first time, which will contribute to accurately analyzing the expression of target genes.

Identification of Knockdown Resistance Mutations in the Cimex hemipterus (Hemiptera: Cimicidae) in Iran.

The worldwide resurgence of tropical bed bug Cimex hemipterus beginning in the late 1990s has led to growing concern. Molecular data on pyrethroid resistance, which is essential for the control strategies, is unknown for C. hemipterus in Iran. The current study evaluated the deltamethrin resistance status of C. hemipterus by bioassay and molecular tests. Live bed bugs were collected from sleeping quarters (dormitories) in the city of Tehran and used for insecticide bioassay tests. For bioassay evaluation, mixed-sex pools of adult bugs were exposed to deltamethrin (0.025%)-treated paper. Polymerase chain reaction assay evaluated resistance-related mutations in the voltage-gated sodium channel gene (VGSC) gene of studied populations. On the basis of the bioassay test within the 48-h exposure to deltamethrin, C. hemipterus were determined to be resistant. Knockdown time ratios (KR50) in the studied populations of C. hemipterus was 5.5-fold compared with those of the C. lectularius Teh strain. DNA sequencing of the VGSC gene revealed the presence of mutations at M918I and L1014 in C. hemipterus. According to the bioassay and molecular results of current study, C. hemipterus showed a high degree of pyrethroid resistance. The application of multiple approaches including physical, biological, and chemical tests should be regarded in future bed bug control strategies.

Morphometrics of the Tropical Bed Bug (Hemiptera: Cimicidae) From Cape Coast, Ghana.

Bed bugs, Cimex lectularius (L.) (Hemiptera: Cimicidae) and Cimex hemipterus (F.), have become established worldwide in recent years largely due to the development of insecticide resistance. However, limited attention has been given to ongoing morphological and macroevolutionary changes within the species and their populations, which could have implications for their control. Here, we evaluated whether bed bugs of the species C. hemipterus inhabiting different communities in Cape Coast, Ghana are undergoing segregation, which could lead to possible speciation. We also aimed to provide a morphometric description of all nymphal stages. Nine-bed bug populations of C. hemipterus were field-collected in Cape Coast and were subjected to geometric morphometric analysis. The multivariate parameters applied distinguished various populations from each of the locations, indicating the presence of morphologically distinct subpopulations of C. hemipterus. Shape-based segregation and shape changes associated with the insect pronotum (which is an important taxonomic character in the Cimicidae) were evident across the populations. Through this comparative study of C. hemipterus, we showed that possible subpopulations of this bed bug are being spread from Ghana. The nymphal stages (first–fifth) of C. hemipterus were distinguished by the length of the last three antennal segment and pronota width; such information contributes to the taxonomic knowledge of the species.

Performance of Pyrethroid-Neonicotinoid Mixture Formulations Against Field-Collected Strains of the Tropical Bed Bug (Hemiptera: Cimicidae) on Different Substrates.

The residual performance of two pyrethroid-neonicotinoid mixture formulations: Temprid SC (10.5% beta-cyfluthrin and 21% imidacloprid) and Tandem (3.5% lambda-cyhalothrin and 11.6% thiamethoxam) on two substrates (glass and filter paper) against eight pyrethroid-resistant strains (BM-MY, BP-MY, CH-MY, GL-MY, KL-MY, SAJ-MY, TT-MY, and QLD-AU) of the tropical bed bug, Cimex hemipterus (F.) (Hemiptera: Cimicidae) collected from Malaysia, and Australia were evaluated. The aging effect of treatment residues on glass was also investigated. A susceptible C. lectularius L. strain (Monheim) was used for comparison. Temprid SC showed varying levels of performance against all C. hemipterus strains: TT-MY (PR50 = 6.5-fold, high performance), BM-MY, GL-MY, SAJ-MY, and QLD-AU (12.8-21.6-fold, moderate performance), BP-MY, and KL-MY (48.2-49-fold, poor performance), CH-MY (128.2-fold, very poor performance). On the other hand, Tandem displayed high performance against all C. hemipterus strains (1.8-8.3-fold). Tandem caused faster mortality than Temprid SC for all strains. Temprid SC and Tandem residues killed C. hemipterus significantly faster on glass than filter paper. Compared with fresh residues, the efficacy of Temprid SC residues significantly declined after one week of aging, while the effectiveness of Tandem residues declined after two weeks of aging. Further investigations using the topical assay method with a diagnostic dose of imidacloprid found two strains (CH-MY and GL-MY) resistant to imidacloprid. The six other strains (BM-MY, BP-MY, KL-MY, SAJ-MY, TT-MY, and QLD-AU) were susceptible.

A Case Study on Tropical Bed Bug, Cimex hemipterus (Hemiptera: Cimicidae) Infestation and Management in Dormitories.

Numerous bed bug research papers have been published in the past 20 yr as a result of bed bug (Cimex spp.) (Hemiptera: Cimicidae) resurgence in the world. Yet, few of them focused on the management of the tropical bed bug, C. hemipterus (F.). Here, we describe a case of tropical bed bug infestation in two dormitory buildings and effectiveness of a tropical bed bug treatment program. The study site consisted of 125 dormitories in two buildings. An initial building-wide monitoring with ClimbUp interceptors revealed 25 infestations. The spatial distribution of bed bug infested rooms showed a significant aggregated distribution pattern with same infestation status for neighboring units sharing walls. All infested rooms were monitored every 2 wk and treated using a combination of steam and diatomaceous earth (DE) dust application if bed bugs were still found. For the 25 initially identified infested rooms, after 14 wk treatment, 44% of them no longer had bed bugs, and the mean number of bed bugs captured per room decreased by 94.1%. A combination of steam and DE dust treatment is an effective strategy for suppressing tropical bed bug infestations in dormitory environment.

Analysis of Transcriptome Difference between Blood-Fed and Starved Tropical Bed Bug, Cimex hemipterus (F.) (Hemiptera: Cimicidae).

Simple SummaryBed bugs are well known for their extreme resilience to starvation. The molecular mechanisms behind this ability, however, are little known. Thus, the whole transcriptomes of blood-fed and starved bed bugs from the species Cimex hemipterus (tropical bed bugs) were sequenced and compared. The transcriptome of tropical bed bugs was initially annotated. Following differentially expressed genes (DEGs) analysis, regulated transcripts were mostly identified in biological processes during blood-feeding and starvation. The results provide an overview of the functional genes proportion of this species and a deeper understanding of the bed bug’s molecular mechanism of resistance to blood feeding and starvation.The reference transcriptome for Cimex hemipterus (tropical bed bug) was assembled de novo in this study, and differential expression analysis was conducted between blood-fed and starved tropical bed bug. A total of 24,609 transcripts were assembled, with around 79% of them being annotated against the Eukaryotic Orthologous Groups (KOG) database. The transcriptomic comparison revealed several differentially expressed genes between blood-fed and starved bed bugs, with 38 of them being identifiable. There were 20 and 18 genes significantly upregulated in blood-fed and starved bed bugs, respectively. Differentially expressed genes (DEGs) were revealed to be associated with regulation, metabolism, transport, motility, immune, and stress response; endocytosis; and signal transduction. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed an enrichment of genes encoding steroid biosynthesis, glycosaminoglycan biosynthesis, butanoate metabolism, and autophagy in both blood-fed and starved bed bugs. However, in blood-fed bed bugs, genes involved in histidine metabolism, caffeine metabolism, ubiquinone/terpenoid-quinone biosynthesis, and sulfur relay system were enriched. On the other hand, starvation activates genes related to nicotinate and nicotinamide metabolism, fatty acid elongation, terpenoid backbone biosynthesis, metabolism of xenobiotics by cytochrome P450, riboflavin metabolism, apoptosis, and protein export. The present study is the first to report a de novo transcriptomic analysis in C. hemipterus and demonstrated differential responses of bed bugs in facing blood-feeding and starvation.

Morphological Identification and DNA Barcoding of the Tropical Bed Bug Cimex hemipterus Fabricius (Hemiptera: Cimicidae) from Thrissur, Kerala, India

Knowledge of bed bugs is scant from Kerala due to improper identification, sparse population and better sanitation. But if an infestation begins it reaches extreme proportions because of the lack of awareness. To the best of our knowledge no detailed scientific studies on the identification of the bed bug species were carried out in Kerala. This study was aimed to perform morphological and molecular identification along with the morphometric analysis of the bed bug species from two sites—Avittappally and Mulankunnathukavu. Morphological identification was made by calculating the width to length ratios of the pronotum and hind femora. DNA barcoding was performed to identify the bed bug species accurately. A total of 21 morphological characters were used to compare bed bugs from the sample and the morphometric characters were measured and analysed using a Leica S8APO stereo microscope. The results indicated that the width-to-length ratios of pronotum and hind femora were 2.2±0.04 mm and 3.5±0.04 mm in Avittappally specimens and 2.1±0.05 mm and 3.2±0.05 mm in Mulankunnathukavu specimens. The genetic distance between the 2 COI sequence of our study were 0.027% and the intraspecific genetic distance between the 2 COI sequence of this study with 5 C. hemipterus sequences reported from other countries was found to be 0.025, 1.227, 0.058, 0.025 and 0.024% for Mulankunnathukavu specimen and 0.001, 1.280, 0.008, 0.001 and 0.001% for Avittappally specimen, respectively. The mean values of HL, HFW, HTW, MTW, FTW between the two places were significantly different. The results of PCA showed that the total percentage of variance reached 89.073% and 93.760%. Based on Morphological and morphometric characteristics along with molecular sequencing we assert that the bed bugs collected from both places were Cimex hemipterus.