Endosymbiont and gut bacterial communities of the brown-banded cockroach, Supella longipalpa.

The brown-banded cockroach (Supella longipalpa) is a widespread nuisance and public health pest. Like the German cockroach (Blattella germanica), this species is adapted to the indoor biome and completes the entirety of its life cycle in human-built structures. Recently, understanding the contributions of commensal and symbiotic microbes to the biology of cockroach pests, as well as the applications of targeting these microbes for pest control, have garnered significant scientific interest. However, relative to B. germanica, the biology of S. longipalpa, including its microbial associations, is understudied. Therefore, the goal of the present study was to quantitatively examine and characterize both the endosymbiont and gut bacterial communities of S. longipalpa for the first time. To do so, bacterial 16S rRNA gene amplicon sequencing was conducted on DNA extracts from whole adult females and males, early instar nymphs, and late instar nymphs. The results demonstrate that the gut microbiome is dominated by two genera of bacteria known to have beneficial probiotic effects in other organisms, namely Lactobacillus and Akkermansia. Furthermore, our data show a significant effect of nymphal development on diversity and variation in the gut microbiome. Lastly, we reveal significant negative correlations between the two intracellular endosymbionts, Blattabacterium and Wolbachia, as well as between Blattabacterium and the gut microbiome, suggesting that Blattabacterium endosymbionts could directly or indirectly influence the composition of other bacterial populations. These findings have implications for understanding the adaptation of S. longipalpa to the indoor biome, its divergence from other indoor cockroach pest species such as B. germanica, the development of novel control approaches that target the microbiome, and fundamental insect-microbe interactions more broadly.

German cockroach extract prevents IL-13-induced CCL26 expression in airway epithelial cells through IL-13 degradation.

Inhaled aeroallergens can directly activate airway epithelial cells (AECs). Exposure to cockroach allergens is a strong risk factor for asthma. Cockroach allergens mediate some of their effects through their serine protease activity; protease activity is also a major contributor to allergenicity. The Th2 cytokine interleukin-13 (IL-13) induces upregulation of the eosinophil chemotactic factor CCL26. CCL26 induces eosinophil migration in allergic inflammation. In this work, we studied the effect of cockroach proteases on IL-13-induced effects. Immersed cultures of the human bronchial epithelial cell line BEAS-2B and air-liquid interface (ALI) cultures of primary normal human bronchial epithelial (NHBE) cells were stimulated with IL-13, Blattella Germanica cockroach extract (CE), or both. IL-13-induced genes were analyzed with qRT-PCR. IL-13 induced upregulation of CCL26, periostin, and IL-13Rα2 in bronchial epithelial cells which were decreased by CE. CE was heat-inactivated (HICE) or pre-incubated with protease inhibitors. HICE and CE preincubated with serine protease inhibitors did not prevent IL-13-induced CCL26 upregulation. CE-degraded IL-13 and specific cleavage sites were identified. CE also decreased IL-4-induced CCL26 upregulation and degraded IL-4. Other serine proteases such as bovine trypsin and house dust mite (HDM) serine proteases did not have the same effects on IL-13-induced CCL26. We conclude that CE serine proteases antagonize IL-13-induced effects in AECs, and this CE effect is mediated primarily through proteolytic cleavage of IL-13. IL-13 cleavage by cockroach serine proteases may modulate CCL26-mediated effects in allergic airway inflammation by interfering directly with the pro-inflammatory effects of IL-13 in vivo.

Intestinal pathogens detected in cockroach species within different food-related environment in Pudong, China.

Cockroaches are considered mechanical transmitters of infectious diseases, posing a threat to human health. This study assessed the potential of cockroaches in food-related environments to mechanically transmit intestinal pathogens. Cockroaches captured with traps were placed together into a low temperature refrigerator at - 80° for 2 h. Standard taxonomic keys and Fluorescent quantitative PCR techniques were applied for species identification and digestive tract etiological examination. A total of 360 cockroach traps were placed, with a positive rate of 20.8%, and 266 cockroaches were captured. In general, compared with other places and areas, the degree of infestation of cockroaches was more serious in catering places and kitchens. Blattella germanica were most found in catering places (40.2%), followed by Periplaneta fuliginosa in schools (22.2%). According to the life stage, among the 128 cockroach samples, 23 were positive for nymphs and 13 were positive for adults. There were statistically significant differences in the intestinal pathogen detection rates between nymphs and adults (P < 0.05). A total of eight intestinal pathogens were detected, and enterovirus infections were the main ones, with sapovirus being the most detected in Blattella germanica or nymph. Shiga toxin-producing Escherichia coli (STEC) was the most frequently isolated bacterium. Blastocystis hominis had the highest isolation rate. In contrast, 12 diarrhoeal disease pathogens were isolated, and the viruses and bacteria with the highest frequencies were norovirus and E. coli, respectively; no parasites were found. Blattella germanica and Periplaneta fuliginosa in food-related environments can act as potential vectors for the spread of intestinal pathogens and may pose a significant threat to public health.

A new amber representative of the genus Ectobius Stephens, 1835 (Blattodea: Ectobiidae) from Storeblt (Denmark).

A new fossil cockroach, Ectobius danekrae sp. nov., is described on the basis of a single male adult individual from late Eocene Danish amber. This is the third representative of the genus Ectobius Stephens, 1835 from Eocene ambers. The new species exhibits plesiomorphic characters of small right stylus and subsymmetrical claws, compared to the known fossil and living species of the genus. The references to Ectobius balticus (Germar et Berendt, 1856) and Margattea Shelford, 1911 sp. presence in Danish amber are insufficient.

Behavioral responses of field-collected German cockroaches to pyrethroids and pyrethroid-formulated insecticides.

BACKGROUND: Pyrethroids are synthetic insecticides with low mammalian toxicity and broad-spectrum activity across insects. One major challenge with pyrethroids is their perceived repellency. This perception can influence decisions made by pest control operators, especially when insecticides are used to reduce insect entry into or movement within structures. One major indoor pest that has been repeatedly shown to be repelled by some pyrethroids is the German cockroach, Blattella germanica. However, most experiments evaluating pyrethroid repellency in the German cockroach have used end-point assays, which do not provide information on the movement that led to the final position. Therefore, we evaluated the kinetic behavioral response of field-collected German cockroaches to five pyrethroid-based products and their active ingredients (A.I.) in open behavioral arenas using advanced video tracking software. In addition, in an effort to compare our free-moving experiments with end-point assays, we evaluated sheltering behavior using two-choice harborage arrestment assays where German cockroaches were provided a choice between pyrethroid-treated and untreated shelters. RESULTS: All pyrethroid-formulated products and their respective A.I.'s failed to affect field-collected German cockroach movement behavior in free-moving assays, while positive controls (DEET, corn mint oil) resulted in reduced time spent by German cockroaches in treated areas. However, despite their willingness to move over pyrethroids-treated surfaces, field-collected German cockroaches displayed a reduced propensity to arrest on pyrethroids treated tents. CONCLUSION: While most pyrethroids/pyrethroid-formulated products affected German cockroach arrestment, pyrethroids and pyrethroid-formulated products failed to change German cockroach movement behavior in free-moving assays. These results indicate the pyrethroids tested act as contact irritants rather than true-spatial repellents on field-collected German cockroaches. This distinction is critical to refining pest management strategies involving pyrethroids. © 2023 Society of Chemical Industry.

A cuticular protein, BgCPLCP1, contributes to insecticide resistance by thickening the cockroach endocuticle.

Overuse of insecticides has led to severe environmental problems. Insect cuticle, which consists mainly of chitin, proteins and a thin outer lipid layer, serves multiple functions. Its prominent role is as a physical barrier that impedes the penetration of xenobiotics, including insecticides. Blattella germanica (L.) is a major worldwide indoor pest that causes allergic disease and asthma. Extensive use of pyrethroid insecticides, including ß-cypermethrin, has selected for the rapid and independent evolution of resistance in cockroach populations on a global scale. We demonstrated that BgCPLCP1, the first CPLCP (cuticular proteins of low complexity with a highly repetitive proline-rich region) family cuticular protein in order Blattodea, contributes to insecticide penetration resistance. Silencing BgCPLCP1 resulted in 85.0 %-85.7 % and 81.0 %-82.0 % thinner cuticle (and especially thinner endocuticle) in the insecticide-susceptible (S) and ß-cypermethrin-resistant (R) strains, respectively. The thinner and more permeable cuticles resulted in 14.4 % and 20.0 % lower survival of ß-cypermethrin-treated S- and R-strain cockroaches, respectively. This study advances our understanding of cuticular penetration resistance in insects and opens opportunities for the development of new efficiently and environmentally friendly insecticides targeting the CPLCP family of cuticular proteins.

Oral toxicity of an artificial sweetener sucralose on the German cockroach (Blattodea: Ectobiidae) and its impact on water balance and gut microbiome.

Artificial or non-nutritive sweeteners are indigestible by most animals. Some sweeteners are orally toxic to insects and have received recent interest as potential safe insecticides due to their low mammalian toxicity. In this study, we investigated the oral toxicity of sucralose on insecticide-susceptible and resistant German cockroaches, Blattella germanica (L.). In a nonchoice test, we evaluated 5, 10, and 20% sucralose solutions. Depending on the cockroach strains, mean mortality ranged from 62.5 to 92.5%, 15 to 55%, and 2.5 to 27.5% for 20, 10, and 5% sucralose, respectively. Next, we measured the impact of a 20% sucralose treatment on water loss rates in the cockroach strains. All strains lost 23.0-30.29% of body water by 6 d. Dehydrated cockroaches were more prone to be killed by sucralose than nondehydrated ones. Lastly, we evaluated the effect of 20% sucralose treatment on gut bacterial composition and found the diversity of gut bacteria in treated cockroaches was significantly reduced after 3 days, implicating a rapid change in the alimentary environment.

Transcriptome analysis revealed that short-term stress in Blattella germanica to ß-cypermethrin can reshape the phenotype of resistance adaptation.

Previous studies on insect resistance have primarily focused on resistance monitoring and the molecular mechanisms involved, while overlooking the process of phenotype formation induced by insecticide stress. In this study, we compared the expression profiles of a beta-cypermethrin (ß-CYP) resistant strain (R) and a susceptible strain (S) of Blattella germanica after ß-CYP induction using transcriptome sequencing. In the short-term stress experiment, we identified a total of 792 and 622 differentially expressed genes (DEGs) in the S and R strains. Additionally, 893 DEGs were identified in the long-term adaptation experiment. To validate the RNA-Seq data, we performed qRT-PCR on eleven selected DEGs, and the results were consistent with the transcriptome sequencing data. These DEGs exhibited down-regulation in the short-term stress group and up-regulation in the long-term adaptation group. Among the validated DEGs, CUO8 and Cyp4g19 were identified and subjected to knockdown using RNA interference. Subsequent insecticide bioassays revealed that the mortality rate of cockroaches treated with ß-CYP increased by 69.3% and 66.7% after silencing the CUO8 and Cyp4g19 genes (P<0.05). Furthermore, the silencing of CUO8 resulted in a significant thinning of the cuticle by 59.3% and 53.4% (P<0.05), as observed through transmission electron microscopy and eosin staining, in the S and R strains, respectively. Overall, our findings demonstrate that the phenotypic plasticity in response to short-term stress can reshape the adaptive mechanisms of genetic variation during prolonged exposure to insecticides. And the identified resistance-related genes, CUO8 and Cyp4g19, could serve as potential targets for controlling these pest populations.

A study of allergen detection panel in Guangzhou, southern China based on real-world data from the past 7 years.

This study aims to reduce the cost of allergen testing for Guangzhou, China by limiting the number of allergens for which patients are tested, and provide a testing panel to improve diagnostic and therapeutic efficiency. This retrospective study of real-world data from 2012 to 2019 included 39,570 patients with suspected allergies in Guangzhou, southern China. All the patients were tested for one or more of the following allergens serum specific immunoglobulin E (sIgE): Dermatophagoides pteronyssinus, Dermatophagoides farinae, cat dander, dog dander, Artemisia vulgaris, Aspergillus fumigatus, Alternaria alternata, Blattella germanica, egg whites, milk, wheat, peanuts, soybeans, Cancer pagurus, and Penaeus monodon by PhadiaCAP 1000. Totally, only the positive rates of allergens sIgE in D. farinae, D. pteronyssinus, milk, egg whites, B. germanica, C. pagurus, A. alternata, and P. monodon were > 10%, the other allergens were between 4-7%. Moreover, among the allergic diseases, dust mites exhibited the overall highest positive rate, followed by milk and B. germanica. In children, milk was the main allergen, whereas in adults, mites, cockroaches, shrimp, and crab allergens had higher positive rates. The optimal scale analysis shows that the multiple sensitization classification of patients can be divided into three categories: I D. farinae and D. pteronyssinus; II. C. pagurus, P. monodon, and B. germanica; III. Milk and egg whites. Generally, a panel including 4 allergens can detect > 90% of the potential allergy in this local population. In Guangzhou, southern China, D. farinae, milk, B. germanica, and A. alternata as a panel screening allergy for suspected allergic patients was suggested base on this study.

Route of pesticide spread on the body surface of Blattella germanica (Linnaeus): a NanoSuit-energy dispersive X-ray spectroscopy analysis.

Numerous studies have focussed on the mechanisms of entry of pesticides into insect body parts such as oral intake, penetration through the integument of the body wall, and inhalation through spiracles. However, little is known about how insecticides spread to the points of entry or the paths on the body surface that are used to reach the target sites. In this study, elemental signals of pesticide-mimicking test solutions were tracked and their routes of spreading in experimental insects (Blattella germanica L.) were investigated using NanoSuit (a method of surface modification) and energy dispersive X-ray spectroscopy, combined with high-resolution scanning electron microscopy. When the test solution initially adhered to the dorsal and/or ventral body surface, it tended to spread horizontally to reach lateral plates. Whereas, when the solution directly adhered to the anterior side of the lateral plates, it spread to posterior segments. In this case, however, spreading in the opposite direction (i.e., the solution directly adhered to the posterior side of the lateral plates) was interrupted at a boundary erected by different groups of fine structures; each protrusion was large, and the arrangement was rather dense in the posterior segments. Morphological features of these fine structures and chemical characteristics of the hydrophobic surface substances potentially regulate the strength of the capillary force, which determines pesticide spreading.

Three new species of Scalida (Blattodea: Blattellidae) from Yunnan, China.

Three new Scalida species from Yunnan are reported: Scalida hamata Qiao & Che sp. nov., Scalida spinosa Qiao & Che sp. nov., Scalida ramiformis Qiao & Che sp. nov.. Morphological descriptions of the new species, including the male genitalia, are provided along with detailed photographs. A list and a key to species of Scalida in the world are provided.

Dual function of the transcription factor Ftz-f1 on oviposition in the cockroach Blattella germanica.

The transcription factor Ftz-f1 has multiple functions in insect development in a spatial-temporal line. One of these roles is in the insect ovaries, specifically in the regulation of steroidogenic enzymes production. We studied the function of F in Blattella germanica oogenesis, as it shows two moments of high expression in ovaries: before the imaginal moult, and just before ovulation in the adult. Injecting dsftz-f1 into adult females, either just after the imaginal moult or just prior to choriogenesis, prevented oviposition, with differences between the two approaches. In 3-day-old adult females treated with dsftz-f1 just after the emergence, the expression of ftz-f1 was not modified, but the steroidogenic genes increased their expression. ftz-f1 transcript levels in the ovaries of 5-day-old dsftz-f1-treated females were significantly depleted, and the expression levels of the same steroidogenic genes began to decrease. These results suggest that Ftz-f1 regulates the expression of steroidogenic genes in B. germanica, with phm possibly being a key target. Ftz-f1 has a different temporal function in the cytoskeleton of follicular cells of the basal ovarian follicles. Early in the gonadotrophic cycle, Ftz-f1 promotes the expression of genes related to the cytoskeleton and muscle proteins, while at the end of the cycle it maintains the expression levels of these genes, thus ensuring correct ovulation.

Differential development and survival of Blattella asahinai and Blattella germanica (Blattodea: Ectobiidae) at six constant temperatures.

The Asian cockroach, Blattella asahinai Mizukubo, is a peridomestic nuisance pest in the southeastern United States. Blattella asahinai is the closest relative to Blattella germanica (L.), the German cockroach, one of the most prolific and widespread domestic pests. Because these two species live in different habitats, they are expected to have differential development patterns reflecting environmental adaptations. Development of B. asahinai and B. germanica cockroach nymphs were observed at six constant temperatures ranging from 10 to 35 °C. At 10 °C and 15 °C, all nymphs died in the first instar, but B. germanica nymphs survived longer (10 °C: 12.9 d; 15 °C: 42.9 d) than B. asahinai nymphs (10 °C 8.2 d; 15 °C 18.4 d) at both temperatures. At 20 °C, 25 °C, and 30 °C, B. asahinai consistently had more instars and longer stadia than B. germanica. At 35 °C, only B. germanica was able to complete nymphal development; cannibalism among B. asahinai nymphs during molting was often observed at this temperature. The results for B. asahinai corroborated previously estimated growth patterns. The lower nymphal development threshold was 14.1 °C for B. germanica and 13.7 °C for B. asahinai. Comparing the development of B. germanica directly with its closest relative reveals specific physiological adaptations that B. germanica has developed for the indoor biome.

Insecticide resistance and its potential mechanisms in field-collected German cockroaches (Blattodea: Ectobiidae) from Thailand.

We investigated insecticide resistance profiles of field populations of the German cockroach, Blattella germanica (L.), collected from central regions of Thailand. Seven strains (PW, RB, MTH, MTS, TL, AY, and SP) were evaluated with diagnostic doses (DD; 3 × LD95 generated from a susceptible strain) of deltamethrin, fipronil, and imidacloprid using topical assays and compared with a susceptible strain (DMSC). Results showed fipronil (2-27% mortality), deltamethrin (16-58% mortality), and imidacloprid (15-75% mortality) resistance in the field strains. Synergism studies with piperonyl butoxide (PBO) and S,S,S-tributyl phosphorotrithioate (DEF) in combination with the DD of insecticides significantly increased (P < 0.05) mortality of the test insects of the field strains suggesting the involvement of P450 monooxygenase and esterase pathways of detoxification. Gel bait evaluations demonstrated that all field-collected strains were resistant to Maxforce Forte (0.05% fipronil), Maxforce Fusion (2.15% imidacloprid), and Advion Cockroach Gel Bait (0.6% indoxacarb) with mean survival times ranging from 1.87-8.27, 1.77-11.72, and 1.19-3.56 days, respectively. Molecular detection revealed that the Rdl mutation was completely homozygous in all field-collected strains except in the PW strain. Field-collected strains were screened for 3 voltage-gated sodium channel (VGSC) mutations associated with pyrethroid resistance. The L993F mutation was present in 5 strains, but no C764R and E434K mutations were detected.

Survival of Salmonella Typhimurium in the hemolymph of the German cockroach vector is limited by both humoral immune factors and hemocytes but not by trehalose metabolism.

The German cockroach (Blattella germanica) has been linked to transmission of Salmonella enterica serovar Typhimurium (S. Typhimurium), but infection dynamics within this vector are poorly characterized. Our recent work has focused on S. Typhimurium infection in the cockroach gut. However, microbial dissemination to the hemolymph is an essential aspect of many vector-borne pathogen transmission cycles and could potentially contribute to S. Typhimurium colonization of cockroaches. Therefore, the goal of this study was to examine the ability of S. Typhimurium to disseminate, survive, and proliferate in the hemolymph of cockroaches after oral infection. We detected only low numbers of bacteria in the hemolymph of a minority of insects (~26%) after oral infection. Further, S. Typhimurium was unable to survive overnight in cell-free hemolymph. Several hypotheses to explain the inability of S. Typhimurium to colonize hemolymph were tested. First, we investigated the ability of S. Typhimurium to metabolize trehalose, the primary sugar in hemolymph. S. Typhimurium grew efficiently in vitro using trehalose as a sole carbon source and mutant strains lacking trehalose metabolism genes exhibited no growth deficiencies in media mimicking the composition of hemolymph, suggesting that trehalose metabolism ability is not a factor involved in restricting survival in hemolymph. On the other hand, heat-inactivated cell-free hemolymph was permissive of S. Typhimurium growth, demonstrating that survival in hemolymph is limited specifically by heat-labile humoral factors. The involvement of cellular immune responses was also investigated and cockroach hemocytes in culture were observed to internalize S. Typhimurium within 1 h of exposure. Most hemocytes harbored few to no bacteria after 24 h, indicating that hemocyte responses are additionally involved in clearing infection from the hemolymph. However, dense intracellular clusters of S. Typhimurium were observed sporadically, suggesting a small subset of hemocytes may serve as reservoirs for bacterial replication. Together, our results reveal that a minute proportion of ingested S. Typhimurium is able to escape the cockroach gut and enter the hemolymph, but this systemic population is limited by both humoral effectors and hemocytes. Thus, we conclude that invasion of the hemolymph appears minimally important for colonization of the cockroach vector and that colonization of the gut is the main driver of vector-borne transmission. Our insight into the antimicrobial mechanisms of cockroach hemolymph also highlights the strong ability of these prevalent pests/vectors to cope with frequent infectious challenges in septic habitats.

Ampicillin treated German cockroach extract leads to reduced inflammation in human lung cells and a mouse model of Asthma.

Cockroaches can cause allergic sensitization in humans via contact with their feces or frass. Antibiotics can affect concentration of major allergen and total bacteria production in German cockroaches (Blattella germanica). This study examined the ability of antibiotic-treated German cockroaches to induce allergic airway inflammation and the effect of antibiotics on their lipopolysaccharide and Bla g1, 2, and 5 expression levels. Specifically, we measured the ability of German cockroach extract (with or without prior antibiotic exposure) to induce allergic inflammation in human bronchial epithelial cells and a mouse model of asthma. Bacterial 16S rRNA and lipopolysaccharide levels were lower in ampicillin-treated cockroaches than in the control group. The Bla g1, Bla g2, and Bla g5 expression in ampicillin-treated cockroaches decreased at both the protein and RNA levels. In human bronchial epithelial cell lines BEAS-2B exposed to the ampicillin-treated extract, expression levels of interleukin-6 and interleukin-8 were lower than that in the control group. The total cell count and eosinophil count in bronchoalveolar lavage fluid was also lower in mice exposed to the ampicillin-treated extract than in those exposed to normal cockroach extract. Mouse lung histopathology showed reduced immune cell infiltration and mucus production in the ampicillin group. Our results showed that ampicillin treatment reduced the symbiont bacterial population and major allergen levels in German cockroaches, leading to reduced airway inflammation in mice. These results can facilitate the preparation of protein extracts for immunotherapy or diagnostics applications.

The prevalence of Wolbachia in multiple cockroach species and its implication for urban insect management.

Cockroach management relies heavily on the use of conventional insecticides in urban settings, which no longer provide the anticipated level of control. Knowledge of cockroach endosymbionts, like Wolbachia, might provide novel avenues for control. Therefore, we screened 16 cockroach species belonging to 3 families (Ectobiidae, Blattidae, and Blaberidae) for the presence of Wolbachia. We mapped the evolution of Wolbachia-cockroach relationships based on maximum likelihood phylogeny and phylogenetic species clustering on a multi-loci sequence dataset (i.e., coxA, virD4, hcpA, and gatB) of Wolbachia genes. We confirmed the previous report of Wolbachia in 1 Ectobiid species; Supella longipalpa (Fab.), and detected the presence of Wolbachia in 2 Ectobiid species; Balta notulata (Stål) and Pseudomops septentrionalis Hebard, and 1 Blaberid species; Gromphadorhina portentosa (Schaum). All cockroach-associated Wolbachia herein detected were clustered with the ancestor of F clade Wolbachia of Cimex lectularius L. (bed bugs). Since Wolbachia provision C. lectularius with biotin vitamins that confer reproductive fitness, we screened the cockroach-associated Wolbachia for the presence of biotin genes. In toto, our results reveal 2 important findings: (i) Wolbachia is relatively uncommon among cockroach species infecting about 25% of species investigated, and (ii) cockroach-associated Wolbachia have biotin genes that likely provide nutritional benefits to their hosts. Thus, we discuss the potential of exploring Wolbachia as a tool for urban insect management.

Wolbachia infection in native populations of Blattella germanica and Periplaneta americana.

Cockroaches are significant pests worldwide, being important in medical, veterinary, and public health fields. Control of cockroaches is difficult because they have robust reproductive ability and high adaptability and are resistant to many insecticides. Wolbachia is an endosymbiont bacterium that infects the reproductive organs of approximately 70% of insect species and has become a promising biological agent for controlling insect pests. However, limited data on the presence or strain typing of Wolbachia in cockroaches are available. PCR amplification and sequencing of the wsp and gltA genes were used to study the presence, prevalence and molecular typing of Wolbachia in two main cockroach species, Blattella germanica (German cockroach) and Periplaneta americana (American cockroach), from different geographical locations of Iran. The Wolbachia endosymbiont was found only in 20.6% of German cockroaches while it was absent in American cockroach samples. Blast search and phylogenetic analysis revealed that the Wolbachia strain found in the German cockroach belongs to Wolbachia supergroup F. Further studies should investigate the symbiotic role of Wolbachia in cockroaches and determine whether lack of Wolbachia infection may increase this insect's ability to tolerate or acquire various pathogens. Results of our study provide a foundation for continued work on interactions between cockroaches, bacterial endosymbionts, and pathogens.

Phylogenetic analysis of Blaberoidea reveals non-monophyly of taxa and supports the creation of multiple new subfamilies.

The superfamily Blaberoidea is a highly species-rich group of cockroaches. High-level blaberoidean phylogenetics are still under debate owing to variable taxon sampling and incongruence between mitochondrial and nuclear evolution, as well as different methods used in various phylogenetic studies. We here present a phylogenetic analysis of Blaberoidea based on a dataset combining the mitochondrial genome with two nuclear markers from representatives of all recognized families within the superfamily. Our results support the monophyly of Blaberiodea, which includes Ectobiidae s.s. (=Ectobiinae), Pseudophyllodromiidae, Nyctiboridae, Blattellidae s.s. (=Blattellinae) and Blaberidae. Ectobiidae s.s. was recovered as sister to the remaining Blaberoidea in all inferences. Pseudophyllodromiidae was paraphyletic with respect to Anaplectoidea + Malaccina. Blattellidae s.s. excluding Anaplectoidea + Malaccina formed a monophyletic group that was sister to Blaberidae. Based on our results, we propose a revised classification for Blaberoidea: Anaplectoidinae subfam.nov. and Episorineuchora gen.nov., and two new combinations at species level within Pseudophyllodromiidae; Rhabdoblattellinae subfam.nov., Calolamprodinae subfam.nov., Acutirhabdoblatta gen.nov., as well as new combinations for three species within Blaberidae. Ancestral state reconstructions based on four morphological characters allow us to infer that the common ancestor of blaberoid cockroaches is likely to be a species with characteristics similar to those found in Ectobiidae, that is, front femur Type B, arolium present, abdomen with a visible gland and male genital hook on the left side.

Glucose- and disaccharide-containing baits impede secondary mortality in glucose-averse German cockroaches.

Glucose aversion in the German cockroach, Blattella germanica (L.), results in behavioral resistance to insecticidal baits. Glucose-averse (GA) cockroaches reject foods containing glucose, even in relatively low concentrations, which protects the cockroaches from ingesting lethal amounts of toxic baits. Horizontal transfer of baits and the resulting secondary mortality have been documented in German cockroaches, including in insecticide resistant strains. However, the effects of the GA trait on secondary mortality have not been investigated. We hypothesized that ingestion of insecticide baits that contain glucose or glucose-containing disaccharides would result in behaviorally relevant glucose levels in the feces, possibly deterring coprophagy by GA nymphs. We fed adult female cockroaches hydramethylnon baits rich in either glucose, fructose, sucrose, or maltose and compared secondary mortality of GA and wild-type (WT) nymphs via coprophagy. When adult females were fed baits containing glucose, sucrose, or maltose and their feces offered to nymphs, secondary mortality was significantly lower in GA nymphs than in WT nymphs. However, survival of GA and WT nymphs was similar on feces generated by adult females fed fructose bait. Analysis of feces indicated that disaccharides in baits were hydrolyzed into glucose, some of which was excreted in the feces of females that ingested the bait. Based on these results, we caution that baits containing glucose or glucose-containing oligosaccharides may impede cockroach interventions; while GA adults and large nymphs avoid ingesting such baits, first instars reject the glucose-containing feces of any WT cockroaches that consumed the bait.