Predictability in the evolution of Orthopteran cardenolide insensitivity.

The repeated evolutionary specialization of distantly related insects to cardenolide-containing host plants provides a stunning example of parallel adaptation. Hundreds of herbivorous insect species have independently evolved insensitivity to cardenolides, which are potent inhibitors of the alpha-subunit of Na+,K+-ATPase (ATPα). Previous studies investigating ATPα-mediated cardenolide insensitivity in five insect orders have revealed remarkably high levels of parallelism in the evolution of this trait, including the frequent occurrence of parallel amino acid substitutions at two sites and recurrent episodes of duplication followed by neo-functionalization. Here we add data for a sixth insect order, Orthoptera, which includes an ancient group of highly aposematic cardenolide-sequestering grasshoppers in the family Pyrgomorphidae. We find that Orthopterans exhibit largely predictable patterns of evolution of insensitivity established by sampling other insect orders. Taken together the data lend further support to the proposal that negative pleiotropic constraints are a key determinant in the evolution of cardenolide insensitivity in insects. Furthermore, analysis of our expanded taxonomic survey implicates positive selection acting on site 111 of cardenolide-sequestering species with a single-copy of ATPα, and sites 115, 118 and 122 in lineages with neo-functionalized duplicate copies, all of which are sites of frequent parallel amino acid substitution. This article is part of the theme issue 'Convergent evolution in the genomics era: new insights and directions'.

Current status and perspectives of fungal entomopathogens used for microbial control of arthropod pests in Brazil.

Entomopathogenic fungi play a central role in Brazil's biopesticide market. Approximately 50% of registered microbial biopesticides comprise mycoinsecticides and/or mycoacaricides consisting of hypocrealean fungi, with most based on Metarhizium anisopliae sensu stricto (s. str.) and Beauveria bassiana s. str. These fungi are mainly used to control spittlebugs in sugarcane fields and whiteflies in row crops, respectively, with annual applications surpassing three million hectares. Research also emphasizes the potential of fungal entomopathogens to manage arthropod vectors of human diseases. Most registered fungal formulations comprise wettable powders or technical (non-formulated) products, with relatively few new developments in formulation technology. Despite the large area treated with mycoinsecticides (i.e., approx. 2 million ha of sugarcane treated with M. anisopliae and 1.5 million ha of soybean treated with B. bassiana), their market share remains small compared with the chemical insecticide market. Nevertheless, several major agricultural companies are investing in fungus-based products with the aim at achieving more sustainable IPM programs for major pests in both organic and conventional crops. Government and private research groups are pursuing innovative technologies for mass production, formulation, product stability and quality control, which will support cost-effective commercial mycoinsecticides. Here, we summarize the status of mycoinsecticides currently available in Brazil and discuss future prospects.

Behavioral responses of pest mole crickets, Neoscapteriscus spp. (Orthoptera: Gryllotalpidae), to selected insecticides.

BACKGROUND: Mole crickets (Neoscapteriscus spp.) consume turfgrasses and pasture grasses and uproot plants by their tunneling, which is detrimental to turf aesthetics and decreases forage quantity and quality. Insecticides are frequently used to prevent damage. In typical field trials, damage symptoms, not percent mortality or achieved level of control, are used to assess treatment efficacy. Here, however, laboratory tests assessed the direct effect of key insecticides on Neoscapteriscus mole cricket behavior. RESULTS: Mole crickets, Neoscapteriscus spp., were able to detect and avoid areas treated with fipronil [formulated product (FP)] and imidacloprid (FP). They tunneled less in sand treated with fipronil and avoided sand treated with fipronil and imidacloprid if given a choice. Mole crickets escaped areas treated with acephate, bifenthrin and fipronil. Bifenthrin and acephate caused increased tunneling during the first 90 min of observation. Fipronil and imidacloprid significantly reduced overall tunneling on treated areas. CONCLUSION: Tested insecticides elicited two types of behavioral changes in Neoscapteriscus mole crickets: increased locomotory activity and tunneling [acephate (organophosphate) and bifenthrin (pyrethroid)] and reduced spatial movement [fipronil (phenylpyrazole) and imidacloprid (neonicotinoid)]. These behavioral responses resulted mainly from contact chemoreception and inherent neurotoxicity of the chemicals on Neoscapteriscus mole crickets. © 2017 Society of Chemical Industry.

Mesoionic pyrido[1,2-a]pyrimidinones: Discovery of triflumezopyrim as a potent hopper insecticide1.

A novel class of mesoionic pyrido[1,2-a]pyrimidinones has been discovered with exceptional insecticidal activity controlling a number of insect species. In this communication, we report the part of the optimization program which led to the discovery of triflumezopyrim as a highly potent insecticide controlling various hopper species. Our efforts in discovery, synthesis, structure-activity relationship elucidation, and biological activity evaluation are also presented.

Selenium accumulation protects plants from herbivory by Orthoptera via toxicity and deterrence.

To investigate whether selenium (Se) accumulation in plants provides a chemical defense against generalist insect herbivores, the feeding preference and performance of a mix of orthopteran species were investigated. The selenium hyperaccumulator Stanleya pinnata and accumulator Brassica juncea were used in herbivory studies in the laboratory, and S. pinnata was also used in a manipulative field experiment. In laboratory studies, both crickets and grasshoppers avoided plants pretreated with selenate, while those given no choice died after eating leaves with elevated Se (447 +/- 68 and 230 +/- 68 microg Se g(-1) DW, respectively). B. juncea has previously been shown to accumulate selenate, while S. pinnata hyperaccumulates methyl-selenocysteine. Thus, these findings demonstrate that both inorganic and organic forms of selenium protect plants from herbivory. Grasshoppers fed S. pinnata contained methylselenocysteine in their midgut and absorbed this form into surrounding tissues. In a manipulative field experiment, methylselenocysteine protected S. pinnata from invertebrate herbivory and increased its long-term survival rate over an entire growth season. * In native habitats of selenium hyperaccumulators, orthopterans represent a major group of insect herbivores. Protection offered by organic selenium accumulation against these herbivores may have promoted the evolution of selenium hyperaccumulation in plants.

Behavioral responses of Schistocerca americana (Orthoptera: Acrididae) to Azadirex (neem)-treated host plants.

Azadirex (azadirachtin and other biologically active extracts from neem trees) has been shown to have considerable potential to be used in integrated pest management systems based on its growth regulator/insecticide properties. Less well known are the antifeedant properties. The feeding-deterrent properties of a commercial azadirex formulation (Azatrol EC) were evaluated using both no-choice and choice tests, the American grasshopper, Schistocerca americana (Drury), and four host plants [savoy cabbage, Brassica oleracea variety capitata L.; cos (romaine) lettuce, Lactuca sativa variety longifolia Lam.; sweet orange, Citrus sinensis variety Hamlin L.; and peregrina, Jatropha integerrima Jacq.]. These studies demonstrated that azadirex application can significantly affect the feeding behavior of grasshoppers. Some degree of protection can be afforded to plants that differ markedly in their innate attractiveness to the insect, although the level of protection varies among hosts. The tendency of grasshoppers to sometimes feed on azadirex-treated foliage suggests that it will be difficult to prevent damage from occurring at all times, on all hosts. No evidence of rapid habituation to azadirex was detected. Rapid loss of efficacy was observed under field conditions, suggesting that daily retreatment might be necessary to maintain protection of plants from feeding.

Laboratory evaluation of Melia azedarach L. and Eucalyptus globulus Labill. Extracts in order to control Ocneridia volxemi Bolivar (Orthoptera, Pamphaginae) hoppers.

This study was carried out to evaluate the effect of Melia azedarach L. (Meliaceae) dry fruit extracts (oil and water) and Eucalyptus globulus Labill. (Myrtaceae) dry leaves extracts (water) on Oneridia volxemi hoppers (fifth-instar) feeding on cereals leaves (Triticum durum) under laboratory conditions. Larva feeding reduced and mortality was significatly (p < 0.05) on cereals leaves sprayed with three concentrations 0, 5, 1 and 2% of oil extracted from Melia azederach. In addition, O. volxemi feeding and mortality was significantly (p < 0.05) on cereals leaves sprayed with two concentrations 50 g/l and 80 g/l of water extracts from Melia azedarach dry fruits soaked for 24 hours. the water extract solution of 80 g/l significantly reduced feeding than the other concentrations 50 and 25 g/l. This study showed also the water extract solution from Eucalyptus globulus dry leaves reduced also feeding and induced the mortality of O. volxemi but not better than Melia azedarach dry fruit extracts water at the same concentration (80 g/l). The aim of this study is in an integrated management program for control of O.

Development of a toxic bait for control of eastern lubber grasshopper (Orthoptera: Acrididae).

This study assessed baits for eastern lubber grasshopper, Romalea guttata (Houttuyn). When offered a choice among several grain-based baits (rolled oats, wheat bran, oat bran, yeast, corn meal, cornflakes) and vegetable oils (canola, corn, peanut, soybean), eastern lubber grasshopper adults preferred bait consisting of wheat bran carrier with corn oil as an added phagostimulant. Other carriers were accepted but consumed less frequently. Discrimination by eastern lubber grasshoppers among oils was poor. Similarly, addition of flavorings (peppermint, anise, lemon, banana) resulted in few significant effects. The carbaryl, wheat bran, and oil bait developed in this study was effective at causing eastern lubber grasshopper mortality in field-cage studies. Significant mortality occurred even though grasshoppers had to locate dishes of bait in a large cage, and could feed on daylilies, or grass growing through the bottom of the cage, rather than on the bran flakes. Consumption of as little as a single carbaryl-treated bran flake could induce mortality, although individuals varied greatly in their susceptibility. The bait matrix developed in this study was readily consumed when in the presence of some plant species. We expect that wheat bran and corn oil bait would be most effective as protection for less preferred plants (tomato, pepper, eggplant, leek, parsley, fennel, daylily, lily of the Nile, and canna lily) because baits were readily consumed in the presence of these plants. Plants that are readily consumed in the presence of bait (preferred plants) included butter crunch lettuce, carrot, yellow squash, cauliflower, collards, green onion, chive, cucumber, cabbage, cantalope, endive, red leaf lettuce, society garlic, caladium, and amaryllis. Baits are likely to be less effective in the presence of such plants. On average, vegetables in Solanaceae (i.e., tomato, pepper, and eggplant) and Apiaceae (i.e., fennel and parsley) elicited high levels of bait-feeding activity, indicating that these vegetables were not highly preferred. The plants tested from Liliaceae, Cucurbitaceae, Asteraceae, and Brassicaceae elicited an intermediate-to-low level of bait feeding.

Unique activity associated with non-insecticidal Bacillus thuringiensis parasporal inclusions: in vitro cell-killing action on human cancer cells.

Parasporal inclusion proteins from a total of 1744 Bacillus thuringiensis strains, consisting of 1700 Japanese isolates and 44 reference type strains of existing H serovars, were screened for cytocidal activity against human leukaemia T cells and haemolytic activity against sheep erythrocytes. Of 1684 B. thuringiensis strains having no haemolytic activity, 42 exhibited in vitro cytotoxicity against leukaemia T cells. These non-haemolytic but leukaemia cell-toxic strains belonged to several H-serovars including dakota, neoleonensis, shandongiensis, coreanensis and other unidentified serogroups. Purified parasporal inclusions of the three selected strains, designated 84-HS-1-11, 89-T-26-17 and 90-F-45-14, exhibited no haemolytic activity and no insecticidal activity against dipteran and lepidopteran insects, but were highly cytocidal against leukaemia T cells and other human cancer cells, showing different toxicity spectra and varied activity levels. Furthermore, the proteins from 84-HS-1-11 and 89-T-26-17 were able to discriminate between leukaemia and normal T cells, specifically killing the former cells. These findings may lead to the use of B. thuringiensis inclusion proteins for medical purposes.

Effect of benserazid and allatectomy on the development and structure of the ovariole in the house cricket, Acheta domestica L.

Light and electron microscopic investigations were carried out on the fourth day imago of the house cricket, Acheta domestica L. treated with benserazid, or allatectomized. Introduction of benserazid (daily on the 1st, 2nd and 3rd days of imaginal life in 0.1 mg/g) failed to influence the development of the oocytes. However, benserazid in the concentration of 0.2 mg/g inhibited the growth of the terminal oocytes, while in 0.6 mg/g it completely blocked the development of terminal oocytes and vitellogenesis. The effect of benserazid proved to be reversible. Application of juvenile hormone analogue (ZR-515) prevented the effect of benserazid on the gonads of the crickets. Ultrastructural damages of the oocytes and follicular epithelium were the same in case of benserazid treatment and allatectomy, although the changes were more expressed in the latter case. The benserazid effect was found to be indirect, caused by damage to the interaction between the aminergic and peptidergic systems of the brain.

Use of the cricket embryo (Acheta domesticus) as an invertebrate teratology model.

Embryos of the cricket Acheta domesticus (L.) have been shown by bioassay to develop gross morphological abnormalities after exposure to a number of complex organic mixtures as well as to display a critical period of teratogen sensitivity and an ability to metabolize xenobiotics during development. Because the assay is simple, inexpensive, short-term (less than two weeks), and objective, it could be useful as an in vivo screen in a hierarchical approach to teratogen detection. Further investigation of cricket embryo responses to known teratogens is needed to establish the predictive value of this assay.

Chemical impurity produces extra compound eyes and heads in crickets.

A chemical impurity isolated from commercially purchased acridine causes cricket embryos to develop extra compound eyes, branched antennae, extra antennae, and extra heads. Purified acridine does not produce similar duplications of cricket heads or head structures nor do the substituted acridines proflavine, acriflavine, or acridine orange. A dose-response relation exists such that the number and severity of abnormalities increase with increasing concentration of the teratogen.

Effect of Insecticide (BHC) on the neurosecretory cells of ventral nerve cord of adult Poekilocercus pictus (Acrididae).

Neurosecretory cells have been observed in all the ganglia of the ventral nerve cord of Poekilocerus pictus. In the suboesophageal ganglion they could be differentiated into 2 types on the basis of their staining reaction, but neurosecretory cells of all the other ganglia stain similarly. Effect of an organic insecticide BHC was observed on the neurosecretory cells of all the locations in the ventral nerve cord. Initially the insecticide acts to trigger the secretory activity but a prolonged action leads to a complete exhaustion of the secretory material which is also accompanied with many cytomorphological alterations such as loss of turgidity and compactness of the cells and appearance of undulations and sometimes complete rupture of the cell wall.

Effect of insecticide (BHC) on brain neurosecretory cells of Poekilocerus pictus (Acrididae).

Neurosecretory cells in the adult males and females of Poekilocerus pictus were studied. They were found in forebrain and hand brain regions in the form of small patches. The median group of pars intercerebralis is however a big patch of about 100...110 cells. The brain neurosecretory cells of Poekilocerus pictus could be differentiated into two types on the basis of their staining reaction i.e. A and B types. The A type cells are restricted to only the median group whereas the other groups have only the B type cells. Effect of 0.01% BHC (LD 50 dose), an organic insecticide was studied. Four incubation periods viz. 1 h, 3 1/2 h, 6 h and 24 hours were tried. the insecticide acted as inducer to the triggering mechanism of enhanced secretory activity and with prolonged incubation a complete exhaustion of secretory material accompanied with lot of disruptions and disturbances to the compactness of the neurosecretory cell wall occurred.

[Effects of reserpine on development of Oedipoda coerulescens L. (Acrididae, Orthoptera) (author's transl)]. / Effets de la réserpine sur le développement d'Oedipoda coerulescens L. (Acridien, Orthoptère).

Inhibitory effects of reserpine on ecdysis and chromatic adaptation in Oedipoda appear to be linked with changes observed in both neural and neuro-endocrine regulating systems.

[Influence of reduced pterine and juvenile hormone on the pigmentation of the phasms Carausius morosus (author's transl)]. / Influence des ptérines réduites et de l'hormone juvénile sur la pigmentation du phasme Carausius morosus

Tetrahydrofolic acid and dimethyltetrahydropterine, FH4 and DMH4P, have an antagonist action. FH4 darkens the phasms. DMH4P and the tryptophane have an inhibitrice influence on the melanisation, probably by the increase of the synthesis or the release of serotonin, in the hypoderm. For the adult phasm remaining ivory, xanthopterin must be added in the treatment. The addition of diapausing chrysalids pieridae extract (containing juvenile hormone JH) and treated by FH4, in the treatment precedent, transmits the pgimentary effect at the first generation, which presents a strong percentage of dark or ivory phasms.