Pathogenic fungi synergistically cooperate with Serratia marcescens to increase cockroach mortality.

The abuse of chemical insecticides has led to strong resistance in cockroaches, and biopesticides with active ingredients based on insect pathogens have good development prospects; however, their slow effect has limited their practical application, and improving their effectiveness has become an urgent problem. In this study, the interaction between Serratia marcescens and Metarhizium anisopliae enhanced their virulence against Blattella germanica and exhibited a synergistic effect. The combination of S. marcescens and M. anisopliae caused more severe tissue damage and accelerated the proliferation of the insect pathogen. The results of high-throughput sequencing demonstrated that the gut microbiota was dysbiotic, the abundance of the opportunistic pathogen Weissella cibaria increased, and entry into the hemocoel accelerated the death of the German cockroaches. In addition, the combination of these two agents strongly downregulated the expression of Imd and Akirin in the IMD pathway and ultimately inhibited the expression of antimicrobial peptides (AMPs). S. marcescens released prodigiosin to disrupted the gut homeostasis and structure, M. anisopliae released destruxin to damaged crucial organs, opportunistic pathogen Weissella cibaria overproliferated, broke the gut epithelium and entered the hemocoel, leading to the death of pests. These findings will allow us to optimize the use of insect pathogens for the management of pests and produce more effective biopesticides.

Differences in Salmonella Typhimurium infection and excretion among laboratory and field strains of the German cockroach suggest a genomic basis for vector competence.

The German cockroach, Blattella germanica, can be a vector of human enteric bacterial pathogens, including Salmonella enterica serovar Typhimurium (S. Typhimurium). Transmission of such pathogens by cockroaches has largely been considered a passive mechanical process, but recent studies have argued against this dogma by demonstrating bacterial proliferation within the cockroach gut and the necessity of specific bacterial genes for successful transmission in the feces, revealing unappreciated biological complexity in the vector-pathogen relationship between cockroaches and S. Typhimurium. However, the influence of naturally occurring variation among cockroach populations on pathogen infection and dissemination has not been investigated. Thus, this study aimed to examine whether distinct strains of B. germanica exhibit differences in their ability to become infected by and disseminate S. Typhimurium. We performed controlled infections of one long-term laboratory strain and three recently field-collected strains reared under identical conditions, then compared bacterial loads in the body and excreta of individual insects. Separately, we also compared rates of necrophagy, a behavior known to contribute to the horizontal spread of S. Typhimurium among cockroaches. Our data show significant differences in infection susceptibility, pathogen shedding in the excreta, and necrophagy between laboratory and field strains as well as between some field strains. These observations represent the first evidence that genomic variation among cockroach populations may influence their ability to become infected by and disseminate pathogens, providing further support for the hypothesis that German cockroaches are active biological vectors rather than passive mechanical vectors of S. Typhimurium. Additional studies are needed to identify the genomic drivers of vector competence for S. Typhimurium in B. germanica.

RNAi of yellow-y, required for normal cuticle pigmentation, impairs courtship behavior and oviposition in the German cockroach (Blattella germanica).

The insect cuticle plays a key role in maintaining the insect's physiological function and behavior. Herein, the yellow-y protein is required to produce black melanin, and is expressed in a pattern that correlates with the distribution of this pigment. However, yellow-y can also have other functions, for instance, in insect behavior, but not much is known. In this study, we have studied the yellow-y gene in one important model and pest species, namely the German cockroach (Blattella germanica), which is to our knowledge the first time reported. In essence, we identified the yellow-y gene (BgY-y) and characterized its function by using RNA interference (RNAi). Silencing of BgY-y gene led to different developmental abnormalities (body weight and wings) in both genders. Specifically, there was an abundant decrease in melanin, turning the body color in pale yellow and the cuticle softer and more transparent. Interestingly, we also observed that the knockdown of BgY-y impaired the male cockroaches to display a weaker response to female-emitted contact sex pheromones, and also that the oviposition ability was weakened in the RNAi females. This study comprehensively analyzed the biological functions of the yellow-y gene in German cockroaches from the perspectives of development, body color, courtship behavior and oviposition, and as a consequence, this may opens new avenues to explore it as a novel pest control gene.

Pinworm Blatticola blattae infection and survival of starving German cockroach Blattella germanica.

The German cockroach Blattella germanica is commonly infected with the pinworm Blatticola blattae. To determine the effect of pinworm infection on cockroach survival, we artificially altered the pinworm infection status of cockroaches and determined the number of survival days under no-feeding conditions, with and without opportunities for fecal feeding. Four lines of the German cockroach (Wn, Wp, Nn, and Np groups) were used in the experiment. Wp and Np were pinworm-infected lines. Wn and Nn were pinworm-free lines. The 50% survival days of cockroaches in the absence of opportunities for fecal feeding were not significantly different in Wp (3.45 days) vs Wn (3.27), and in Np (4.60) vs Nn (4.48). In contrast, in the presence of fecal feeding, the 50% survival times for the pinworm-infected Wp (4.04) and Np (6.65) were significantly longer than those for the pinworm-free Wn (2.77) and Nn (5.46). The number of survival days without feeding was significantly higher in the pinworm-infected group given the opportunity to eat feces than in the non-infected group. These results suggest that pinworm infection of cockroaches during starvation, in association with fecal feeding, may be associated with longer survival.

Two Lactones Non-toxic to Humans Are Effective Repellents for the German Cockroach.

Lactones are cyclic esters of hydroxy carboxylic acids, present in several fruits and animals consumed by humans. There is evidence that some lactones modify insect behavior. The aim of the present study was to evaluate the repellent effect of four lactones (γ- and δ-nonalactone, and γ- and δ-dodecalactone) in first instar nymphs of the German cockroach, Blattella germanica (Linnaeus). To assess repellency, a nymph was placed on a circle of filter paper, half of which had been treated with lactone dissolved in acetone and the other half with acetone alone. The behavior of the nymph was recorded and the time the nymph spent in each half of the paper was quantified using Ethovision XT 10.1 software. Values of Distribution Coefficient (DC) were calculated: DC = (Tt - TA) / Tt, where Tt is the experimental time and TA is the time the nymph spent in the area treated with the repellent agent. DC can vary between 0 and 1. Values significantly higher than 0.5 indicate repellency. N,N-diethyl-meta-toluamide (DEET) was used as a positive control. DEET, δ- and γ-nonalactone caused repellency as from 77.9 µg/cm2, whereas γ- and δ-dodecalactone had a repellent effect starting at 779.0 µg/cm2. The values of DC for these concentrations were 0.89 (DEET), 0.86 (γ-nonalactone), 0.87 (δ-nonalactone), 0.83 (γ-dodecalactone), and 0.72 (δ-dodecalactone). To our knowledge, this is the first report of repellency produced by lactones in the German cockroach. This work allowed to identify two lactones that have a repellent effect similar to DEET.

Acetyl coenzyme A carboxylase modulates lipogenesis and sugar homeostasis in Blattella germanica.

Lipid and sugar homeostasis is critical for insect development and survival. In this study, we characterized an acetyl coenzyme A carboxylase gene in Blattella germanica (BgACC) that is involved in both lipogenesis and sugar homeostasis. We found that BgACC was dominantly expressed in the fat body and integument, and was significantly upregulated after molting. Knockdown of BgACC in 5th-instar nymphs did not affect their normal molting to the next nymphal stage, but it caused a lethal phenotype during adult emergence. BgACC-RNA interference (RNAi) significantly downregulated total free fatty acid (FFA) and triacylglycerol (TAG) levels, and also caused a significant decrease of cuticular hydrocarbons (CHCs). Repression of BgACC in adult females affected the development of oocytes and resulted in sterile females, but BgACC-RNAi did not affect the reproductive ability of males. Interestingly, knockdown of BgACC also changed the expression of insulin-like peptide genes (BgILPs), which mimicked a physiological state of high sugar uptake. In addition, BgACC was upregulated when B. germanica were fed on a high sucrose diet, and repression of BgACC upregulated the expression of the glycogen synthase gene (BgGlyS). Moreover, BgACC-RNAi increased the circulating sugar levels and glycogen storage, and a longevity assay suggested that BgACC was important for the survival of B. germanica under conditions of high sucrose uptake. Our results confirm that BgACC is involved in multiple lipid biogenesis and sugar homeostasis processes, which further modulates insect reproduction and sugar tolerance. This study benefits our understanding of the crosstalk between lipid and sugar metabolism.

The Response of Susceptible and Pyrethroid-Resistant Blattella germanica (Dyctioptera: Blattellidae) to Shelter-Associated Cues.

In this work, it was studied the role of faeces in the location and permanence in a shelter in susceptible and pyrethroid-resistant individuals of Blattella germanica (Linnaeus 1767). Additionally, the effect of different concentrations of palmitic acid on the modulation of these behaviours was tested. A shelter constituted by a square cardboard structure was offered to susceptible as well as to resistant specimens. The shelter bases were treated with faecal extracts obtained from susceptible or resistant cockroaches, or with solutions of palmitic acid. The behaviour of susceptible as well as resistant specimens was analysed using infrared videography software. Susceptible's faecal extract attracted both specimens since the time spent by cockroaches to locate the treated shelters was lower, whereas the faecal extract from resistant insects did not elicit any effect on both strains. Faecal extracts showed an arrestant effect on both strains, suggested by the time spent inside the shelter that was significantly higher in their presence. On the other hand, treatment with palmitic acid produced an attractant or a repellent effect depending on the concentration and strain. The tested lower concentration was attractant to susceptible insects, but did not produce any effect on resistant ones. In addition, the higher concentrations did not produce any effect on susceptible individuals, but resulted repellent for resistant ones. Palmitic acid did not produce an arrestant effect on the strains as there was not an increase in time spent inside the shelter in the presence of this substance. An increase in the number of visits to the shelter and to the periphery was also observed in shelters treated with the faecal extract and with the lower concentration of palmitic acid. These results show that compounds of the susceptible faeces were attractant to cockroaches of both strains, while faecal extracts from resistant insects were not. Moreover, a dual effect of palmitic acid was observed, being attractant at low concentrations and repellent as concentration increased. Additionally, a difference in the concentration threshold at which the effect of this substance changes was observed between strains.

Gustatory polymorphism mediates a new adaptive courtship strategy.

Human-imposed selection can lead to adaptive changes in sensory traits. However, rapid evolution of the sensory system can interfere with other behaviours, and animals must overcome such sensory conflicts. In response to intense selection by insecticide baits that contain glucose, German cockroaches evolved glucose-aversion (GA), which confers behavioural resistance against baits. During courtship the male offers the female a nuptial gift that contains maltose, which expediates copulation. However, the female's saliva rapidly hydrolyses maltose into glucose, which causes GA females to dismount the courting male, thus reducing their mating success. Comparative analysis revealed two adaptive traits in GA males. They produce more maltotriose, which is more resilient to salivary glucosidases, and they initiate copulation faster than wild-type males, before GA females interrupt their nuptial feeding and dismount the male. Recombinant lines of the two strains showed that the two emergent traits of GA males were not genetically associated with the GA trait. Results suggest that the two courtship traits emerged in response to the altered sexual behaviour of GA females and independently of the male's GA trait. Although rapid adaptive evolution generates sexual mismatches that lower fitness, compensatory behavioural evolution can correct these sensory discrepancies.

Effects of copper and zinc oxide nanoparticles on German cockroach development, indoxacarb resistance, and bacterial load.

BACKGROUND: The German cockroach, Blattella germanica, is a ubiquitous and medically significant urban pest. The ongoing development of insecticide resistance in global populations of B. germanica has complicated control efforts and created a need for improved tools. We previously reported that disruption of the gut microbiota by oral administration of the antimicrobial doxycycline reduced resistance in an indoxacarb resistant field strain and also delayed nymphal development and reduced adult fecundity. However, the application of doxycycline for cockroach control in the field is impractical. Here, we sought to determine whether two metal nanoparticles with known antimicrobial properties, copper (Cu) and zinc oxide (ZnO), have similar effects to doxycycline on the physiology of B. germanica and could provide more practical alternatives for control. RESULTS: We found that dietary exposure to 0.1% Cu nanoparticles, but not ZnO, significantly delays the development of nymphs into adults. However, neither of the nanoparticles altered the fecundity of females, and ZnO surprisingly increased resistance to indoxacarb in a resistant field strain, in contrast to doxycycline. Semi-quantitative polymerase chain reaction (qPCR) further revealed that prolonged dietary exposure (14 days) to Cu or ZnO nanoparticles at the low concentration readily consumed by cockroaches (0.1%) does not reduce the load of the bacterial microbiota, suggesting alternative mechanisms behind their observed effects. CONCLUSIONS: Together, our results indicate that ingestion of Cu nanoparticles can impact German cockroach development through an undetermined mechanism that does not involve reducing the overall load of the bacterial microbiota. Therefore, Cu may have some applications in cockroach control as a result of this activity but antagonistic effects on insecticide resistance should be considered when evaluating the potential of nanoparticles for cockroach control. © 2023 Society of Chemical Industry.

Microbe-mediated activation of indoxacarb in German cockroach (Blattella germanica L.).

The German cockroach (Blattella germanica L.) is a major urban pest worldwide and is notorious for its ability to detoxify and resist insecticides. German cockroaches have generalist feeding habits that expose them to a range of potential hazardous substances and host a wide variety of unique microbial species, which may potentially facilitate unique detoxification capabilities. Since field German cockroach populations are routinely exposed to both bait and spray insecticide treatments, we hypothesized whether these unique gut microbes might play roles in toxicological processes of the host insect. The goals of this research were to understand the metabolic processes inside the German cockroach gut after treatment with kanamycin, a broad-ranging antibiotic, and indoxacarb, an oxadiazine pro-insecticide used in cockroach bait products. In these experiments, two resistant cockroach strains were obtained from field populations in Danville, IL and compared to a susceptible laboratory strain that had no previous exposure to insecticides (J-wax). Roaches provided kanamycin-infused water had lower median mortality to indoxacarb compared to the control treatment in feeding bioassays regardless of strain, but in vial (surface contact) bioassays, only susceptible cockroaches experienced a shift in mortality apparently due to their greater susceptibility. When frass extracts of indoxacarb-fed cockroaches were analyzed, less of the active, hydrolytic metabolite DCJW (N-decarbomethoxyllated JW062) was produced relative to the parent compound indoxacarb with the antibiotic KAN. This result was further corroborated by hydrolase activity assays of whole homogenized cockroach guts. Taken together these results provide novel evidence of microbe-mediated pro-insecticide activation in the cockroach gut.

Detection of Voltage-Gated Sodium Channel (VGSC) Mutation in Deltamethrin-Resistant Blattella germanica L. from Indonesia.

<b>Background and Objective:</b> German cockroach (<i>Blattella germanica</i> L.) is one of the most common residential pests in Indonesia. Controlling the population face obstacles due to insecticide resistance, especially to deltamethrin. This research investigated the resistance status and the possibility of a Voltage-Gated Sodium Channel (VGSC) knockdown resistance mutation (L1014F) in two field strains of German cockroaches collected from two cities in Indonesia (Bukittinggi, named RMKN-BKT and Bandung, named KRSA-BDG) with VCRU-WHO as the standard strain. <b>Materials and Methods:</b> This study started with a bioassay test to determine the lethal dose of 50% (LD₅₀) for each strain and followed by a molecular test for mutation detection. <b>Results:</b> The results showed that the RMKN-BKT and KRSA-BDG strains were highly resistant to deltamethrin with RR50 values of 80,090 times and 73,272 times, respectively. Only the RMKN-BKT strain was shown to carry L1014F kdr mutation which lead to an amino acid replacement from leucine (TTG) to phenylalanine (TTC). Two silent mutations were also found in both field strains at codons 983 (TGC/cysteine→TGT/cysteine) and 984 (GGG/ glycine→GGA/glycine) which were suggested as polymorphism phenomena. The absence of the L1014F mutation in the Bandung strain does not exclude the possibility of the presence of the VGSC mutation at other points. <b>Conclusion:</b> It requires subsequent investigation in mutation detection at other points and the possible presence of other resistance mechanisms to get a precise solution in the population control. Bioinsecticides may stand as a breakthrough so that the strategy will no longer focus on insecticides.

An aggregation model of cockroaches with fast-or-slow motion dichotomy.

We propose a mathematical model, namely a reaction-diffusion system, to describe social behaviour of cockroaches. An essential new aspect in our model is that the dispersion behaviour due to overcrowding effect is taken into account as a counterpart to commonly studied aggregation. This consideration leads to an intriguing new phenomenon which has not been observed in the literature. Namely, due to the competition between aggregation towards areas of higher concentration of pheromone and dispersion avoiding overcrowded areas, the cockroaches aggregate more at the transition area of pheromone. Moreover, we also consider the fast reaction limit where the switching rate between active and inactive subpopulations tends to infinity. By utilising improved duality and energy methods, together with the regularisation of heat operator, we prove that the weak solution of the reaction-diffusion system converges to that of a reaction-cross-diffusion system.

Multiple Mechanisms Confer Fipronil Resistance in the German Cockroach: Enhanced Detoxification and Rdl Mutation.

Populations of Blattella germanica (L.) (German cockroach) have been documented worldwide to be resistant to a wide variety of insecticides with multiple modes of action. The phenylpyrazole insecticide fipronil has been used extensively to control German cockroach populations, exclusively in baits, yet the highest reported fipronil resistance is 38-fold in a single population. We evaluated five populations of German cockroaches, collected in 2018-2019 in apartments in North Carolina and assayed in 2019, to determine the status of fipronil resistance in the state. Resistance ratios in field-collected strains ranged from 22.4 to 37.2, indicating little change in fipronil resistance over the past 20 yr. In contrast, resistance to pyrethroids continues to escalate. We also assessed the roles of detoxification enzymes in fipronil resistance with four synergists previously shown to diminish metabolic resistance to various insecticides in German cockroaches-piperonyl butoxide, S,S,S-tributyl phosphorotrithioate, diethyl maleate, and triphenyl phosphate. These enzymes appear to play a variable role in fipronil resistance. We also sequenced a fragment of the Rdl (resistant to dieldrin) gene that encodes a subunit of the GABA receptor. Our findings showed that all field-collected strains are homozygous for a mutation that substitutes serine for an alanine (A302S) in Rdl, and confers low resistance to fipronil. Understanding why cockroaches rapidly evolve high levels of resistance to some insecticides and not others, despite intensive selection pressure, will contribute to more efficacious pest management.

Actividad insecticida de aceites esenciales sobre Blatella germanica (Linneaus, 1767) (Dictyoptera: Blattellidae) / nsecticidal activity of essential oils against Blatella germanica (Linneaus, 1767) (Dictyoptera: Blattellidae)

Blattella germanica (Linneaus, 1767) es una especie de cucaracha considerada plaga de la salud pública por estar asociada a gran número de microorganismos causantes de enfermedades al hombre. Para su control se utilizan diferentes tipos de formulaciones a base de insecticidas sintéticos a los cuales en su gran mayoría es resistente. En este contexto existe un interés creciente por los insecticidas botánicos. En el siguiente trabajo se evaluaron los aceites de Citrus aurantium (L.,1753), Ocimum basilicum (L.,1753), Piper aduncum subsp ossanum (C.DC. Saralegui) y Eucalyptus globulus (Labill, 1800) mediante aplicación tópica de un microlitro en el primer esternito abdominal de los individuos. Los cuatro aceites mostraron actividad insecticida sobre adultos de B. germanica con CL50 que oscilaron entre 58 µg/µL para O. basilicum y 250 µg/µL para P. aduncum(AU)´

Blattella germanica (Linneaus, 1767) is a cockroach species considered a public health pest, since it is associated with a great number of disease-causing microorganisms in humans. For its control, different types of synthetic-based insecticidal formulations are used, to which it is mostly resistant. In this context, there is a growing interest in botanical insecticides. In this research, oils from Citrus aurantium (L., 1753), Ocimum basilicum (L., 1753), Piper aduncum subsp. ossanum (C.DC. Saralegui), and Eucalyptus globulus (Labill, 1800) were evaluated by topical application of 1 µL to the first abdominal sternum of the individuals. The four essential oils evaluated showed insecticidal activity against adult B. germanica with LC50 ranging from 58µg/µL for O. basilicum to 250µg/µL for P. aduncum(AU)´

Antimicrobial peptide expression in the cockroach gut during enterobacterial infection is specific and influenced by type III secretion.

Omnivorous synanthropic cockroaches, such as the German cockroach (Blattella germanica), are reservoirs and vectors of enteric bacterial pathogens. A lifestyle conducive to frequent encounters with high loads of diverse bacteria may have led to the evolution of unique innate immune systems in these insects. The innate immune response of insects relies largely on generalized mechanisms to sense and eliminate foreign microbes. However, analyses of the genomes of common synanthropic cockroaches previously revealed a repertoire of pathogen associated molecular pattern (PAMP) receptors and antimicrobial peptides (AMPs) that is significantly expanded relative to most holometabolous insect models and vectors, supporting the intriguing possibility that cockroaches may encode enhanced recognition within their immune system and may possess an enhanced capacity to fine tune innate immune responses. Investigating how cockroaches respond to infection with enterobacteria provides the opportunity to expand our fundamental knowledge of the regulation of insect innate immunity in a context that is biologically and medically relevant. German cockroaches can harbor both Salmonella enterica serovar Typhimurium and Escherichia coli in their gut without experiencing pathogenesis. The former colonizes the gut and replicates while the latter persists only transiently. We hypothesized that differences in the innate immune response may contribute to or result from the difference in infection dynamics between the two enterobacteria. To test this hypothesis, we used qRT-PCR to analyze expression of five genes encoding representative AMPs (Attacins, Blattellicin, Defensins) in the gut of German cockroaches 1 and 24 h after ingestion of live or heat-killed enterobacteria. We found that robust AMP expression was induced in response to ingestion of a live wild-type strain of S. Typhimurium, but not in response to live E. coli, heat-killed S. Typhimurium, or a live mutant strain of S. Typhimurium lacking type III secretion systems. These results indicate that the cockroach immune system does not respond to stimulation with high levels of ingested bacterial PAMPs such as peptidoglycan. Rather, AMP expression in the gut appears to be induced by active bacterial colonization involving type III secretion. We speculate that this form of regulation may have evolved to prevent over activation of the immune system from frequent ingestion of innocuous, non-colonizing, or non-viable bacteria. While additional work is needed to delineate the molecular mechanisms underlying our observations, our findings provide significant novel insight into the immunological adaptation of cockroaches to life in septic environments as well as the factors that regulate bacterial pathogen transmission by these insects.

Prevalence of Cockroaches, Bed Bugs, and House Mice in Low-Income Housing and Evaluation of Baits for Monitoring House Mouse Infestations.

Pests are frequently found in homes, especially in apartment buildings in low-income communities. We investigated the prevalence and patterns of pest infestations in low-income communities in four cities (Jersey City, Linden, Paterson, Trenton) in New Jersey, USA. Resident interviews, visual inspections, and the placement of monitors were used to identify pest infestations. A total of 1,753 apartments from 19 buildings or building complexes were accessed. The infestation rates of cockroaches, bed bugs (Cimex lectularius L.), and house mice (Mus musculus domesticus Schwarz and Schwarz) were 37, 9, and 20%, respectively. Among apartments with cockroaches, 97.8, 2.5, and 0.8% had German cockroach [Blattella germanica (L.)], American cockroach [Periplaneta americana (L.)], and Oriental cockroach (Blatta orientalis L.), respectively. The percentage of residents who were aware of the presence of cockroaches, bed bugs, and house mice was 70.8, 55.3, and 56.8%, respectively. The prevalence of pest infestation was associated with resident ethnicity and gender. Among 856 interview responses, 78% implemented methods themselves to control pests in their homes in the past six months. Chocolate spread detected 99% of house mouse infestations and was much more sensitive than three commercial blank baits. Significant differences were also observed in the feeding preference of the three commercial blank baits.

Genome-wide identification and expression pattern analysis of novel chemosensory genes in the German cockroach Blattella germanica.

The German cockroach Blattella germanica is an important urban insect pest worldwide. In many insects, chemosensation is essential for guiding their behaviors for survival. Although a large number of chemosensory-related genes have been identified in B. germanica, little information on tissue-specific and developmental expression patterns has not been uncovered yet. In this study, we performed transcriptome analysis of different B. germanica tissues to reveal novel chemosensory proteins (CSPs) and sensory neuron membrane proteins (SNMPs). In addition, a phylogenetic tree and gender-specific expression of multiple chemosensory gene families have been analyzed. We identified three CSPs genes (BgerCSP11, BgerCSP12, and BgerCSP13) and five SNMP genes in B. germanica. Tissue-specific expression profiling showed that CSP1, 8, and 9 exhibited significant expression levels in both adult and 5th instar nymph antennae. The results have paved the way for further functional study of the chemosensory mechanism in B. germanica and provided potential insecticide targets.

Melanin synthesis genes BgTH and BgDdc affect body color and cuticle permeability in Blattella germanica.

Melanin is involved in cuticle pigmentation and sclerotization of insects, which is critical for maintaining structural integrity and functional completeness of insect cuticle. The 2 key enzymes of tyrosine hydroxylase (TH) and dopa decarboxylase (DDC) predicted in melanin biosynthesis are usually conserved in insects. However, it is unclear whether their function is related to epidermal permeability. In this study, we identified and cloned the gene sequences of BgTH and BgDdc from Blattella germanica, and revealed that they both showed a high expression at the molting, and BgTH was abundant in the head and integument while BgDdc was expressed highest in the fat body. Using RNA interference (RNAi), we found that knockdown of BgTH caused molting obstacles in some cockroaches, with the survivors showing pale color and softer integuments, while knockdown of BgDdc was viable and generated an abnormal light brown body color. Desiccation assay showed that the dsBgTH-injected adults died earlier than control groups under a dry atmosphere, but dsBgDdc-injected cockroaches did not. In contrast, when dsRNA-treated cockroaches were reared under a high humidity condition, almost no cockroaches died in all treatments. Furthermore, with eosin Y staining assay, we found that BgTH-RNAi resulted in a higher cuticular permeability, and BgDdc-RNAi also caused slight dye penetration. These results demonstrate that BgTH and BgDdc function in body pigmentation and affect the waterproofing ability of the cuticle, and the reduction of cuticular permeability may be achieved through cuticle melanization.

Metarhizium anisopliae is a valuable grist for biocontrol in beta-cypermethrin-resistant Blattella germanica (L.).

BACKGROUND: The widespread use of chemical insecticides has resulted in the development of resistance in German cockroaches worldwide, and biopesticides based on entomopathogenic fungi as active ingredients have become a promising alternative strategy. Resistance can change many of the physiological and biochemical characteristics of insect pests, such as cuticle thickness, detoxification enzyme activity, and even intestinal flora composition. Thus, potential interactions between pathogenic fungi and insecticide resistance may lead to unpredictable changes in pest susceptibility to fungi. RESULTS: Beta-cypermethrin-resistant German cockroaches were more susceptible to infection with the fungus Metarhizium anisopliae regardless of age and sex. Histopathological results showed that the infection of resistant strains (R) by M. anisopliae was visibly faster than that of susceptible strains (S). The gut microbiota of the S strain indicated a stronger ability to inhibit fungi in vitro. The abundance of Parabacteroides, Lachnoclostridium, and Tyzzerella_3 decreased significantly in the R strain, and most demonstrated the ability to regulate glucose and lipid metabolism, and antifungal infections. The expression levels of Akirin, BgTPS, and BgPo genes in the R strain were significantly lower than those in the S strain, while BgChi and CYP4G19 gene expression were significantly higher. The mortality of cockroaches infected with M. anisopliae decreased to varying degrees after RNA interference, reflecting the role of these genes in antifungal infection. CONCLUSIONS: Results confirmed that insecticide resistance may enhance cockroach susceptibility to fungi by altering intestinal flora and gene expression. Fungal biopesticides have high utilization value in pest control and insecticide resistance management strategies. © 2021 Society of Chemical Industry.

The different aspects of attractive toxic baits containing fipronil for control of the German cockroach (Blattella germanica).

The use of Attractive Toxic Baits (ATBs) is considered to be a low-risk practical method for controlling cockroaches. This study evaluates the attractiveness of a lab-made, fipronil-containing bait, and its effect on the food consumption and mortality of the German cockroach Blattella germanica, under field and laboratory conditions. Different developmental stages of the cockroach were used to determine their preferred carbohydrate/protein rich foods and examine the effectiveness of lab-made baits. The analysis of variance (ANOVA) with Tukey's Test was determined using SAS 9.1 and GraphPad Prism software programs. The significance level was considered at p<0.05. The most-consumed foods were carbohydrate-rich foods (biscuit and banana powder) and food consumption rate was highest in non-gravid females. The most attractive bait ingredients were 20% roasted peanut butter, 50% biscuit and 30% banana powder. The highest lethality was recorded from the baits containing 0.02% fipronil under laboratory conditions, while infested houses baited with the lab-made bait showed 76.5% and 100% decline, respectively, in cockroach numbers in the first- and fourth-week post-baiting. In conclusion, palatable foods with a pleasant odour, like biscuit powder, banana powder and peanut butter were the most effective ingredients for the ATBs. The ATBs impregnated with 0.02% fipronil provide a promising approach for control of the German cockroach. However, there is a need to evaluate the potentials of the lab-made baits, under laboratory and field conditions, in the control of other health-important cockroaches.