Insecticidal Activity of Essential Oils and Their Synergistic Effect on Improving the Efficacy of ß-Cypermethrin against Blattella germanica.

We screened the contact activity of 32 commercial essential oils (EOs) and their synergistic effect with ß-cypermethrin against Blattella germanica. Results showed that the most effective EOs against B. germanica were from Illicium verum, Syzygium aromaticum, and Cinnamomum camphora, with LD50 values of less than 500 µg/insect. The most potent synergistic effects of ß-cypermethrin on B. germanica were from Dysphania ambrosioides and Mentha canadensis. Both oils have a co-toxic factor of 133.33. The results of the major compound testing of the EOs showed that trans-anisaldehyde and thymol have the best insecticidal activity against B. germanica, with LD50 values of 141.30 and 138.61 µg/insect, respectively. The compounds with the best synergistic effect on ß-cypermethrin were γ-terpinene and linalool at a concentration of 0.5%. The co-toxic factors for γ-terpinene and linalool were 150 and 133.33, respectively, which were similar to the synergistic effect observed with 2% piperonyl butoxide.

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.

Effects of solid and aqueous dietary diflubenzuron ingestion on some biological parameters in synthetic pyrethroid-resistant German cockroach, Blattella germanica L. (Blattodea: Ectobiidae).

Cockroaches, widespread pests found in metropolitan areas, are known as vectors of various disease agents, including viruses, fungi and antibiotic-resistant bacteria, as well as causing allergies in humans. Insect growth regulators have been used in pest management for several decades. These insecticides disrupt insect development and reproduction. Chitin synthesis inhibitors interfere with chitin biosynthesis in insects, causing abortive moulting and mortality, as well as inhibiting egg fertility, and larval hatching in insects. In this research, we evaluated the various effects of diflubenzuron, a chitin synthesis inhibitor, on synthetic pyrethroid-resistant German cockroach (Blattella germanica L. Blattodea: Ectobiidae), including ootheca production, oothecal viability, ootheca incubation time, the number of nymphs emerging from the ootheca and survivorship of nymphs. The cockroaches were fed diets that contained diflubenzuron, which was added to solid bait (impregnated fish food) and ingestible aqueous bait (impregnated cotton). Three concentrations (0.5%, 1% and 2%) were used in the experiments. As a result, diflubenzuron treatment led to ootheca production ranging from 60% to 100%; statistically, no difference was found between the treatment and the control groups. The number of nymphs emerging from the first and second ootheca was reduced by 40%-100% in the diflubenzuron-treated groups compared with the control. Nymphs exposed to diflubenzuron-impregnated solid bait and ingestible aqueous bait experienced mortality exceeding 92.1% and 66.27% within 15 days, respectively. In conclusion, diflubenzuron is a potential insecticide for use in cockroach baits to control B. germanica, as it caused high nymphal and embryonic mortality in the synthetic pyrethroid-resistant population and decreased the number of nymphs emerging from the ootheca.

Effects of Nicotine and Tobacco-Related Products on the Feeding Behavior of the German Cockroach (Blattodea: Blattellidae).

Animals use olfaction to detect developmentally significant volatile organic compounds (VOCs) in their local environment. As part of a wider study aiming to demonstrate that the olfactory responses of animals to VOCs can be modified through the creation of a drug-addicted status and association with a selected VOC, we investigated nicotine and tobacco smoke particulate (TSP) extract as possible addictive compounds for male German cockroaches, Blattella germanica (Linnaeus). In feeding experiments using an artificial food stimulus, food treated with TSP extract was preferred over untreated food. Surprisingly, nicotine, which was expected to be the most important addictive tobacco component, did not induce noticeable effects on cockroach behavior. Both TSP extract and nicotine were shown to be phagostimulants. Olfactometry assays that measured odor-mediated insect behavior demonstrated that male B. germanica did not choose TSP-extract-treated food even when attempts were made specifically to train them via this modality. These results support a hypothesis that B. germanica needs to consume TSP-containing food to show a clear preference for this stimulus and that gustatory mechanisms are involved due to compounds present in the TSP extract.

Resistance development characteristics of reared German cockroach (Blattodea: Blattellidae) to chlorpyrifos.

Understanding the process of resistance development of German cockroach, Blattella germanica (L.), in detail is necessary to potentially delay the development of insecticides resistance by rotation or discontinuation of insecticides at the right time. In this study, we investigated the resistance development of the reared German cockroach to chlorpyrifos (CPF) for 23 generations from susceptible cockroaches. CPF 50% lethal dose (LD50) and resistance ratio of each generation cockroaches were determined. The CPF LD50 to each generation cockroaches was used as the insecticide selection pressure of this generation by topical application. The resistance development curve was depicted according to the CPF LD50 to all 23 generations of cockroaches. As a result, a highly resistant German cockroach cohort to CPF, which the resistance ratio was 21.63, was obtained after 23 generations' selection. During the selection, the cockroaches developed low resistance from F1 to F5, moderate resistance from F6 to F12, and high resistance from F13 to F23. There was a rapid resistance increase every 5-7 generations. The resistance growing showed relatively slow from F1 to F11. The fastest growing phase of the resistance was from F12 to F20, in which accounted for more than 80% of the total resistance increase in 23 generations. The development of resistance to CPF tended to slow down from F21 to F23. These findings may provide a basis for the rational use of insecticides, delaying the development of resistance by rotation or discontinuation.

Insecticidal application of essential oils loaded polymeric nanoparticles to control German cockroach: Design, characterization and lethal/sublethal effects.

Essential oils (EO) from peppermint, palmarosa, geranium, lavender and rosemary were tested against the German cockroach, Blatella germanica L. (Blattaria: Blatellidae). Peppermint and palmarosa oils were the most effective and were included in a polyethylene glycol 6000 matrix to obtain EO loaded polymeric nanoparticles (EOPN). The physicochemical analyses indicated that, at 7 days postformulation, peppermint EOPN had sizes of 380 nm, the loading efficiency (LE) was 72.25% and the polydispersity index (PDI) was >0.4 (polydisperse sample). Palmarosa EOPN had sizes of 191 nm; LE was 89.75% and PDI was <0.25 (monodisperse sample). Peppermint and palmarosa EOPN enhanced the lethal and sublethal effects of the EO on B. germanica. These results suggest that the newly developed nanoinsecticides could be successfully used to control German cockroach.

Topical Toxicity Profiles of Some Aliphatic and Aromatic Essential Oil Components Against Insecticide-Susceptible and Resistant Strains of German Cockroach (Blattodea: Ectobiidae).

Toxicity profiles of four aliphatic (α-pinene, cyclononanone, limonene, nerolidol), four aromatic (ß-thujaplicin, carvacrol, eugenol, tropolone) essential oil components (EOCs), and permethrin were investigated against three strains of German cockroach, Blattella germanica (L.). The strains include a susceptible strain (S), and two multi-resistant strains - strains D and E. Also, a synergism bioassay, using piperonyl butoxide (PBO) was conducted. The most toxic EOCs were aromatic EOCs carvacrol, eugenol, and tropolone, followed by aliphatic EOC limonene; all had LD50 values of <0.7 mg/µl. Four of the EOCs were equally toxic against all the strains, with carvacrol being the most toxic, followed by eugenol, tropolone, and α-pinene. The other four EOCs were more toxic against strain S than against the two resistant strains. Permethrin was significantly more toxic to strain S (LD50 = 0.056 µg/µl) compared with the resistant strains (D = 2.138 µg/µl, E = 1.730 µg/µl). Toxicity of aliphatic EOCs correlated positively with their molecular weight against strain E only, whereas both molecular weight and vapor pressure of aromatic EOCs correlated significantly with toxicity in all strains. Strain D exhibited the greatest resistance (RR of 6.7) to EOCs, and synergism to the aliphatic EOC cyclononanone. Clear synergism with PBO was observed in permethrin against resistant strains, but not in all of the EOCs, suggesting multiple resistance mechanisms in the resistant cockroaches. These findings give insight on the potential of EOCs to be incorporated as parts of an IPM approach to managing insecticide resistant German cockroaches.

Life history and functional capacity of the microbiome are altered in beta-cypermethrin-resistant cockroaches.

Cockroaches are widely perceived to evolve resistance to insecticides. Over-expression of a resistance-conferring gene can be costly and may require energy and resource reallocation for metabolic and developmental processes. To evaluate whether changes in the composition of gut microbiota in Blattella germanica affected its resistance evolution to beta-cypermethrin and to determine the role of gut microbiota in host growth and development, we studied the relationship between insecticide resistance and the diversity and genetic content of gut microbiota in cockroaches. Results suggest beta-cypermethrin-resistant cockroaches (R strain) exhibited a delayed development period and reduced adult longevity compared with susceptible cockroaches (S strain). Based on 16S rRNA gene sequencing and community metagenomics, we found that the relative abundance of Lactobacillus and Acetobacteraceae were significantly lower in the R strain compared with the S strain in the foregut and midgut of both strains. Functional annotation of Kyoto Encyclopedia of Genes and Genomes (KEGG) modules of midgut genes in the two strains revealed that 10.6% were involved in metabolism, while the relative abundance in the R strain was 7.4%. Unigenes were also translated into amino acid sequences and assigned to protein families based on hits to the Carbohydrate-Active enzymes (CAZy) database. This process identified the glycoside hydrolases, glycosyl transferases and carbohydrate-binding modules of the S strain as all being significantly higher in diversity than those in the R strain. Overall, we conclude that fitness-related costs increased in the resistant strain of cockroaches compared with the susceptible strain, and the variation in insect gut microbiota, especially those related to growth and development, was an important influencing factor.

Exposure risks and ineffectiveness of total release foggers (TRFs) used for cockroach control in residential settings.

BACKGROUND: The German cockroach, Blattella germanica, is one of the most challenging pests to eradicate from indoor environments. Professional pest control is often prohibitively expensive, prompting low-income residents to turn to over-the-counter consumer products, including total release foggers (TRFs, "bug bombs"). Despite their widespread use, little is known regarding either the associated pesticide exposure risks or the efficacy of TRFs. METHODS: Cockroach-infested homes were recruited into the study. Wipe samples were collected from various surfaces before TRFs were discharged, immediately after, and one month later to determine pesticide exposure risks in 20 homes (divided equally among four different TRF products). Simultaneously, cockroach populations were monitored in all homes to assess the efficacy of TRFs. In parallel, 10 homes were treated with gel baits (divided equally between two bait products), to compare TRFs to a more targeted, low-risk, do-it-yourself intervention strategy. RESULTS: TRFs failed to reduce cockroach populations, whereas similarly priced gel baits caused significant declines in the cockroach populations. Use of TRFs resulted in significant pesticide deposits throughout the kitchen. Across all products, pesticides, and horizontal kitchen surfaces, pesticide residues following TRF discharge were 603-times (SEM ±184) higher than baseline, with a median increase of 85 times. CONCLUSIONS: The high risks of pesticide exposure associated with TRFs combined with their ineffectiveness in controlling German cockroach infestations call into question their utility in the marketplace, especially because similarly priced and much safer bait products are highly effective in the indoor environment.

Disruption of the microbiota affects physiological and evolutionary aspects of insecticide resistance in the German cockroach, an important urban pest.

The German cockroach, Blatella germanica, is a common pest in urban environments and is among the most resilient insects in the world. The remarkable ability of the German cockroach to develop resistance when exposed to toxic insecticides is a prime example of adaptive evolution and makes control of this insect an ongoing struggle. Like many other organisms, the German cockroach is host to a diverse community of symbiotic microbes that play important roles in its physiology. In some insect species, there is a strong correlation between the commensal microbial community and insecticide resistance. In particular, several bacteria have been implicated in the detoxification of xenobiotics, including synthetic insecticides. While multiple mechanisms that mediate insecticide resistance in cockroaches have been discovered, significant knowledge gaps still exist in this area of research. Here, we examine the effects of altering the microbiota on resistance to a common insecticide using antibiotic treatments. We describe an indoxacarb-resistant laboratory strain in which treatment with antibiotic increases susceptibility to orally administered insecticide. We further reveal that this strains harbors a gut microbial community that differs significantly from that of susceptible cockroaches in which insecticide resistance is unaffected by antibiotic. More importantly, we demonstrate that transfer of gut microbes from the resistant to the susceptible strain via fecal transplant increases its resistance. Lastly, our data show that antibiotic treatment adversely affects several reproductive life-history traits that may contribute to the dynamics of resistance at the population level. Together these results suggest that the microbiota contributes to both physiological and evolutionary aspects of insecticide resistance and that targeting this community may be an effective strategy to control the German cockroach.

Repellency of zerumbone identified in Cyperus rotundus rhizome and other constituents to Blattella germanica.

The compounds 1,8-cineole and zerumbone (ZER) from the Cyperus rotundus rhizome along with another 11 previously identified rhizome essential oil constituents and α-humulene, which lacks the only carbonyl group present in ZER, as well as binary mixtures of ZER and seven active compounds were tested for repellency to male B. germanica. The results were compared to N,N-diethyl-3-methylbenzamide (deet). In filter-paper choice tests, ZER was the most repellent compound, and α-humulene was ineffective, which indicates that the α,ß-unsaturated carbonyl group of ZER is a prerequisite component for repellency. At 81.5 µg cm-2, enhanced repellency was produced by binary mixtures of ZER and 1,8-cineole, (+)-dihydrocarvone or (R)-(+)-limonene (70:30, 50:50 and 30:70 ratios by weight). These mixtures were very effective against male B. germanica within 24 h and were more repellent than a single compound or deet alone. The optimum ZER content was determined to be more than 50%. In Ebeling choice box tests at 652.4 µg cm-2, these compounds and deet resulted in complete repellency to intact male B. germanica, while they exhibited 35-47% repellency to antennectomized male one. Mixtures formulated from the active constituents of the C. rotundus rhizome could be useful as potential repellents for controlling B. germanica.

Hydronopylformamides: Modification of the Naturally Occurring Compound (-)-ß-Pinene to Produce Insect Repellent Candidates against Blattella germanica.

The development of a novel repellent plays an important role in the integrated control of Blattella germanica. A series of novel hydronopylformamides derivatives were synthesized from a naturally occurring compound (-)-ß-pinene. The structures of these hydronopylformamides derivatives were characterized by Fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance spectroscopy (¹H-NMR and 13C-NMR), and electron impact mass spectrometry (EI-MS). Repellency of these hydronopylformamides derivatives against Blattella germanica was evaluated by the using petri dish arena method. The results showed that four derivatives (compounds 8a, 8b, 8c and 8e) exhibited repellency against Blattella germanica at a concentration of 20 mg/mL. Compound 8a was the most active compound among these derivatives, where the repelling ratios of compound 8a against Blattella germanica were 66.10%, 50.46%, 48.26%, at concentrations of 20 mg/mL, 10 mg/mL, and 5 mg/mL, respectively. In addition, compound 8a showed better repellency than the traditional insect repellent N, N-diethyl-3-methylbenzamide (DEET), which indicated that compound 8a had a good application prospect in the prevention of Blattella germanica. This research hopes to promote the value-added utilization of (-)-ß-pinene and the development of novel German cockroach repellents.

Unidirectional Cross-Resistance in German Cockroach (Blattodea: Blattellidae) Populations Under Exposure to Insecticidal Baits.

Insect pests, including the German cockroach, Blattella germanica (L.), are prone to the development of physiological resistance when exposed to a number of insecticide sprays, and cross-resistance is frequently observed. Toxic baits are often used as a primary method of controlling German cockroaches, also resulting in heavy selection pressure from insecticidal baits. In response to this pressure, cockroach populations have developed aversion to specific inert ingredients in bait. Here, we examined the effect of exposure to baits containing fipronil, indoxacarb, or hydramethylnon on the development of physiological resistance to the same and other insecticides in a number of German cockroach strains. We found that prolonged exposure to baits containing fipronil or indoxacarb increased physiological resistance to these compounds. However, no increase in physiological resistance against any insecticide was observed in response to exposure to hydramethylnon bait. Additionally, we found that exposure to fipronil bait increased cross-resistance to indoxacarb. On the other hand, exposure to indoxacarb bait did not increase cross-resistance to fipronil. Neither fipronil nor indoxacarb bait exposure increased resistance to hydramethylnon. Interestingly, the development of insecticide resistance in response to bait exposure was strain-dependent and influenced by bait palatability. Our results demonstrate that exposure to toxic baits, particularly those containing fipronil, plays a significant role in the development of insecticide resistance, including cross-resistance, in German cockroaches. Further, although insecticide resistance in response to baits is mediated by exposure through the oral route, the molecular mechanisms at play are likely different for each insecticide.

Persistence of a sugar-rejecting cockroach genotype under various dietary regimes.

Glucose-aversion is a heritable trait that evolved in a number of German cockroach (Blattella germanica L.) populations in response to strong selection with glucose-containing insecticide baits. However, in the absence of glucose-containing bait, glucose-averse (GA) cockroaches have lower performance than wild-type (WT) cockroaches in several fitness-determining traits. We allocated 48 caged populations initiated with homozygous GA and WT adults to four dietary treatments consisting of either pure rodent chow, rodent chow mixed to yield a content of either 20% glucose or 20% fructose, or a treatment consisting of choice between the 20% glucose- and the 20% fructose-containing food. After 6 months we found significantly higher frequency of WT individuals in populations restricted to the 20% glucose food, and after 12 months all dietary treatments contained significantly more WT individuals than expected. In accompanying experiments, we found lower survival and longer development time of GA nymphs restricted to glucose-containing food. We furthermore found evidence for assortative mating of females with males from their own genotype, with significant differences within WT cockroaches. Our study shows experimental evidence that within heterogeneous populations, WT German cockroaches will over time prevail in abundance over GA individuals, even when glucose is not a dietary component.

Development of Diagnostic Insecticide Concentrations and Assessment of Insecticide Susceptibility in German Cockroach (Dictyoptera: Blattellidae) Field Strains Collected From Public Housing.

Insecticide resistance in German cockroaches (Blattella germanica (L.)) has been a barrier to effective control since its first documentation in the 1950s. A necessary first step toward managing resistance is to understand insecticide susceptibility profiles in field-collected strains so that active ingredients (AIs) with lowest resistance levels can be identified. As a first step in this study, diagnostic concentrations (DCs) were determined for 14 insecticide AIs based on lethal concentrations that killed 99% or 90% of the individuals from a susceptible lab strain (JWax-S). Next, cockroaches were collected from two low-income multifamily housing complexes in Danville, IL, and Indianapolis, IN, and used to establish laboratory strains. These strains were screened against the 14 AI-DCs in vial bioassays, and susceptibility profiles were determined by comparing percent mortalities between the field strains relative to the JWax-S strain. Results revealed lowest resistance of field strains to boric acid, abamectin, dinotefuran, clothianidin, thiamethoxam, and chlorfenapyr. For the AIs hydramethylnon and imidacloprid, field strains did not display survivorship different than the lab strain, but >90% mortality was never achieved. Lastly, both field strains displayed resistance to indoxacarb, fipronil, acetamiprid, beta-cyfluthrin, bifenthrin, and lambda-cyhalothrin, but at varying levels. These results satisfy two objectives. First, baseline monitoring DCs were established for 14 insecticides presently registered for use against cockroaches, which represents a useful resource. Second, our findings reveal insecticide AIs with lowest resistance levels for use in forthcoming field studies that will investigate impacts of different insecticide deployment strategies on resistance management and evolution in cockroach field populations.

Insecticide Resistance of Several Field-Collected German Cockroach (Dictyoptera: Blattellidae) Strains.

German cockroaches, Blattella germanica (L.), remain one of the most difficult indoor insect species to control because of its ability to develop resistance to insecticides. The toxicity and resistance levels of five technical-grade insecticides (permethrin, chlorpyrifos, propoxur, imidacloprid, and fipronil) were determined for adult males of seven strains of the German cockroach, a laboratory-reared susceptible strain (S) and six field-collected strains (B, D, E, G, H, and I). Using topical application methods, fipronil was the most toxic insecticide to all seven strains. The LD50 values of fipronil in the susceptible strain (S) and the field-collected strains B, D, E, G, H, and I were 1.33, 2.62, 11.53, 5.07, 7.66, 5.15, and 10.15 ng/insect, respectively. The field-collected strains were most resistant to permethrin among the five insecticides, except for strain H. The resistance ratios of strains B, D, E, G, and I to permethrin were 31.8, 37.3, 51.9, 34.9, and 37.5, respectively. With a resistance ratio of 6.4, the field-collected strain H was most resistant to chlorpyrifos. The field-collected strains were not significantly resistant to propoxur. Strains B, H, and I were not significantly resistant to imidacloprid when compared with the susceptible strain. Based on the different resistance ratios for each insecticide, we conclude that there are high rates of insecticide resistance in German cockroaches from Franklin County, NC, and that the field-collected strains most likely had different treatment histories.

Repellent Activity of Apiaceae Plant Essential Oils and their Constituents Against Adult German Cockroaches.

We evaluated the repellent activity of 12 Apiaceae plant essential oils and their components against male and female adult German cockroaches, Blattella germanica L., to find new natural repellents. Of all the plant essential oils tested, ajowan (Trachyspermum ammi Sprague) and dill (Anethum graveolens L.) essential oils showed the most potent repellent activity against male and female adult German cockroaches. Repellent activities of chemicals already identified in active oils were also investigated. Of the compounds identified, carvacrol, thymol, and R-(-)-carvone showed >80% repellent activity against male and female adult German cockroaches at 2.5 µg/cm2. S-(+)-Carvone, (+)-dihydrocarvone, and terpinen-4-ol showed >70% repellent activity against male and female adult German cockroaches at 10 µg/cm2. Our results indicated that Apiaceae plant essential oils and their constituents have good potential as natural repellents against adult German cockroaches.

Increased Expression of P-Glycoprotein Is Associated With Chlorpyrifos Resistance in the German Cockroach (Blattodea: Blattellidae).

A principal method for control of the German cockroach, Blattella germanica (L.), is the broad-spectrum organophosphorus insecticide, chlorpyrifos (O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphorothioate); however, extensive and repeated application has resulted in the development of resistance to chlorpyrifos in this insect. Evidence suggests that ATP-binding cassette protein transporters, including P-glycoprotein, are involved in insecticide resistance. However, little is known of the role of P-glycoprotein in insecticide resistance in the German cockroach. Here, we developed a chlorpyrifos-resistant strain of German cockroach and investigated the relationship between P-glycoprotein and chlorpyrifos resistance using toxicity assays; inhibition studies with two P-glycoprotein inhibitors, verapamil and quinine; P-glycoprotein-ATPase activity assays; and western blotting analysis. After 23 generations of selection from susceptible strain cockroaches, we obtained animals with high resistance to chlorpyrifos. When P-glycoprotein-ATPase activity was inhibited by verapamil and quinine, we observed enhanced susceptibility to chlorpyrifos in both control and chlorpyrifos-resistant cockroaches. No significant alterations of P-glycoprotein expression or ATPase activity were observed in cockroaches acutely exposed to LD50 doses of chlorpyrifos for 24 h, while P-glycoprotein expression and ATPase activity were clearly elevated in the chlorpyrifos-resistant cockroach strain. Thus, we conclude that P-glycoprotein is associated with chlorpyrifos resistance in the German cockroach and that elevated levels of P-glycoprotein expression and ATPase activity may be an important mechanism of chlorpyrifos resistance in the German cockroach.

Molecular basis of the remarkable species selectivity of an insecticidal sodium channel toxin from the African spider Augacephalus ezendami.

The inexorable decline in the armament of registered chemical insecticides has stimulated research into environmentally-friendly alternatives. Insecticidal spider-venom peptides are promising candidates for bioinsecticide development but it is challenging to find peptides that are specific for targeted pests. In the present study, we isolated an insecticidal peptide (Ae1a) from venom of the African spider Augacephalus ezendami (family Theraphosidae). Injection of Ae1a into sheep blowflies (Lucilia cuprina) induced rapid but reversible paralysis. In striking contrast, Ae1a was lethal to closely related fruit flies (Drosophila melanogaster) but induced no adverse effects in the recalcitrant lepidopteran pest Helicoverpa armigera. Electrophysiological experiments revealed that Ae1a potently inhibits the voltage-gated sodium channel BgNaV1 from the German cockroach Blattella germanica by shifting the threshold for channel activation to more depolarized potentials. In contrast, Ae1a failed to significantly affect sodium currents in dorsal unpaired median neurons from the American cockroach Periplaneta americana. We show that Ae1a interacts with the domain II voltage sensor and that sensitivity to the toxin is conferred by natural sequence variations in the S1-S2 loop of domain II. The phyletic specificity of Ae1a provides crucial information for development of sodium channel insecticides that target key insect pests without harming beneficial species.

Evaluation of sublethal effects of polymer-based essential oils nanoformulation on the german cockroach.

The German cockroach, Blattella germanica (L.), is a serious household and public health pest worldwide. The aim of the present study was to evaluate the sublethal activity of polymer-based essential oils (EOs) nanoparticles (NPs) on adults of B. germanica. The LC50 and LC25 for contact toxicity were determined. To evaluate the repellency of EOs and NPs at LC25, a software was specially created in order to track multiple insects on just-recorded videos, and generate statistics using the obtained information. The effects of EOs and NPs at LC25 and LC50 on the nutritional physiology were also evaluated. The results showed that NPs exerted sublethal effects on the German cockroach, since these products enhance the repellent effects of the EOs and negatively affected the nutritional indices and the feeding deterrence index.