Susceptibility of Eucalyptus trees to defoliation by the Eucalyptus snout beetle, Gonipterus sp. n. 2, is enhanced by high foliar contents of 1,8-cineole, oxalic acid and sucrose and low contents of palmitic and shikimic acid.

Gonipterus sp. n. 2 (Coleoptera, Curculionidae) is an invasive, commercially important weevil that causes large-scale defoliation of Eucalyptus trees. The weevil specifically feeds on young leaves and new shoots, thus reducing tree growth. The weevil displays a very strong preference for certain Eucalyptus genotypes, however, this behaviour and the chemistry underlying it is poorly understood, thereby complicating the selection of resistant trees. To elucidate the feeding preference of Gonipterus sp. n. 2, we assessed the relative levels of susceptibility of 62 Eucalyptus genotypes from 23 species using a laboratory choice assay. This revealed large intraspecific variation in susceptibility to weevil feeding, which for certain species, exceeded the interspecific variation. A semiquantitative metabolite profile analysis on 13 genotypes revealed strong correlations of 10 metabolites to feeding damage. The behavioural effects of the identified compounds were assessed through an in vitro feeding preference assay using artificial diets as well as under field conditions. This revealed three phagostimulants (1,8-cineole, oxalic acid and sucrose) and two feeding deterrent compounds (shikimic acid and palmitic acid) for Gonipterus sp. n. 2. These chemical markers can be applied to tree breeding programmes for the selection of resistant genotypes to reduce damage caused by Gonipterus weevils.

Synthesis and activity of 3-oxo-α-ionone analogs as male attractants for the solanaceous fruit fly, Bactrocera latifrons (Diptera: Tephritidae).

A series of 3-oxygenated α-ionone analogs have been developed as highly specific male lures for the solanaceous fruit fly Bactrocera latifrons, a pest of solanaceous fruits. We compared the attractant and phagostimulant activities of analogs with or without (i) unsaturations at the 4,5- and/or 7,8-positions and (ii) oxygen moieties at the 3- and/or 9-positions of the ionone molecule. Since naturally occurring vomifoliol (V2) was found to induce a highly potent phagostimulant activity in B. latifrons males, related analogs including dehydrovomifoliol (V1), 6-hydroxy-α-ionone (U1), and 6-hydroxy-α-ionol (U2) were synthesized to evaluate their attractant and phagostimulant activities. Synthetic V1, V2, U1, and U2 exhibited low attractant activity, but their phagostimulant activity was relatively high. Optical isomers of 3-oxo-7,8-dihydro-α-ionone (P3) and V1 were prepared to examine the stereochemical specificity of attractants. (+)-(6R)-P3 and (+)-(6S)-V1 exhibited the corresponding activities, while their respective antipodal enantiomers were found entirely inactive.

Sucrose Improves Insecticide Activity Against Drosophila suzukii (Diptera: Drosophilidae).

The addition of sucrose to insecticides targeting spotted wing drosophila, Drosophila suzukii (Matsumura), enhanced lethality in laboratory, semifield, and field tests. In the laboratory, 0.1% sucrose added to a spray solution enhanced spotted wing drosophila feeding. Flies died 120 min earlier when exposed to spinosad residues at label rates enhanced with sucrose. Added sucrose reduced the LC50 for dried acetamiprid residues from 82 to 41 ppm in the spray solution. Laboratory bioassays of spotted wing drosophila mortality followed exposure to grape and blueberry foliage and/or fruit sprayed and aged in the field. On grape foliage, the addition of 2.4 g/liter of sugar with insecticide sprays resulted in an 11 and 6% increase of spotted wing drosophila mortality at 1 and 2 d exposures to residues, respectively, averaged over seven insecticides with three concentrations. In a separate experiment, spinetoram and cyantraniliprole reduced by 95-100% the larval infestation of blueberries, relative to the untreated control, 7 d after application at labeled rates when applied with 1.2 g/liter sucrose in a spray mixture, irrespective of rainfall; without sucrose infestation was reduced by 46-91%. Adding sugar to the organically acceptable spinosyn, Entrust, reduced larval infestation of strawberries by >50% relative to without sugar for five of the six sample dates during a season-long field trial. In a small-plot field test with blueberries, weekly applications in alternating sprays of sucrose plus reduced-risk insecticides, spinetoram or acetamiprid, reduced larval infestation relative to the untreated control by 76%; alternating bifenthrin and phosmet (without sucrose) reduced infestation by 65%.

Blood constituents as phagostimulants for the bed bug Cimex lectularius L.

Many hematophagous arthropods are stimulated by blood constituents to initiate feeding. We used a membrane-based feeding system to identify chemicals that stimulate acceptance and engorgement responses in various life stages of bed bugs. Water was fortified with a variety of compounds (e.g. salts, amino acids, vitamins, nucleotides, cholesterol and fatty acids) in these bioassays. ATP was the most effective phagostimulant in adults and nymphs, resulting in >70% of bed bugs fully engorging. Addition of NaCl to low ATP solutions that alone elicited <50% engorgement significantly enhanced feeding responses of bed bugs. A comparison of feeding responses with solutions of various adenine nucleotides showed that ATP was more stimulatory than ADP, which was more effective than AMP. Feeding assays with physiological levels of other blood constituents such as d-glucose, albumin, globulin, cholesterol and mixtures of vitamins and amino acids did not stimulate engorgement, suggesting that adenine nucleotides are the most important feeding stimulants in bed bugs. Identification of phagostimulants for bed bugs will contribute towards the development of artificial diets for rearing purposes, as well as for the development of alternative methods to eliminate bed bug infestations.

Do Phagostimulants, Alone or Combined with Ammonium Acetate, Di-Ammonium Phosphate, and Acetic Acid, Effectively Attract Both Sexes of Peach Fruit Flies, Bactrocera zonata (Diptera: Tephritidae)?: Insights from Laboratory and Field Bioassays.

Laboratory and field assays of three sets of experiments were conducted to evaluate the impact of different phagostimulants alone and in combination with other phagostimulant lure sources, such as ammonium acetate, DAP, and acetic acid, on the attractancy of both sexes of B. zonata. In the first experiment, the laboratory olfactometer study revealed that out of eleven phagostimulants, banana, mulberry, mango, guava, molasses, and protein hydrolysate exhibited moderate attractancy (15.2-60.2%) to B. zonata. Unexpectedly, banana and protein hydrolysate were demonstrated to be highly attractive phagostimulants for starved female B. zonata (53.6% and 60.2%, respectively). In the field study, none of the tested phagostimulants exhibited high attractancy; however, banana, mulberry, protein hydrolysate, guava, mango, and molasses demonstrated moderate attractancy (5.6-35.6%) to B. zonata. In the second experiment, out of five phagostimulant-mixtures, phagostimulant-mixture-4 proved highly attractive (40.5-68.6% and 45.5-51.2%), followed by phagostimulant-mixture-3, which proved to be moderately attractive (17.0-22.5% and 28.4-36.1%) to B. zonata in olfactometer and field studies, respectively. In the third experiment, out of five phagostimulant-AdMixtures, phagostimulant-AdMixture-4 demonstrated strong attractiveness in the olfactometer (41.6-68.7%) and field studies (52.7-58.7%) for B. zonata, while the rest of the AdMixtures demonstrated moderate to no attractiveness for B. zonata. So, phagostimulant-AdMixture-4 with GF-120 could be used in the development of a phagostimulant bait station which attracts the maximum B. zonata population and ultimately provides pest-free fruits to the farmers.

Characteristics of a Spray-Dried Porcine Blood Meal for Aedes aegypti Mosquitoes.

Research into mosquito-borne illnesses faces hurdles because feeding fresh animal blood to rear female mosquitoes presents logistical, economic, and safety challenges. In this study, a shelf-stable additive (spray-dried porcine blood; SDPB) hypothesized to supply accessible hemoglobin was evaluated within an alternative meal (AM) containing whey powder and PBS for rearing the yellow fever mosquito Aedes aegypti. LC-MS/MS proteomics, microbial assays, and particle reduction techniques confirmed and characterized the functionality of hemoglobin in SDPB, while engorgement, fecundity, egg viability, and meal stability bioassays assessed AM performance. Chemical assays supported hemoglobin as the phagostimulant in SDPB with aggregates partially solubilized in the AM that can be more accessible via particle reduction. Unpaired two-tailed t-tests indicate that the AM stimulates oogenesis (t11 = 13.6, p = 0.003) and is stable under ambient (1+ y; t12 = 0.576, p = 0.575) and aqueous (14 d; t12 = 0.515, p = 0.639) conditions without decreasing fecundity. Egg hatch rates for the ninth generation of AM-reared Ae. aegypti were 50-70+%. With further development, this meal may serve as a platform for mass rearing or studying effects of nutritional additives on mosquito fitness due to its low cost and stability. Future work may examine tuning spray drying parameters and resulting impacts on hemoglobin agglomeration and feeding.

Sugar Bait Composition Containing Ivermectin Affect Engorgement and Mortality of the Mosquito Aedes aegypti (Diptera: Culicidae).

Toxins and attractants have been studied and used in sugar baits for mosquitoes. However, little importance has been given to the combination of sugar concentration, bait composition, and their relationship with mosquito engorgement and mortality. Therefore, the present study evaluated the effects of three concentrations of sucrose on baits with and without an attractant (concentrated guava juice), on engorgement and mortality rates of adult Aedes aegypti (Linnaeus, Diptera: Culicidae). Toxic sugar baits (TSB) and attractant toxic sugar baits (ATSB) containing 10, 50, and 70% sucrose and 100 ppm ivermectin (IVM) were prepared to assess engorgement and mortality rates. Subsequently, different concentrations of IVM (0.312-100 ppm) in TSB and ATSB were prepared with sucrose concentrations of 10 and 70% to determine the lethal concentrations (LC50 and LC90) values. Engorgement on the baits was observed under a stereomicroscope, and mortality was followed up to 48 h after feeding. In general, more mosquitoes engorged on TSB regardless of the sugar concentration, while higher concentrations of sugar in ATSBs resulted in higher numbers of mosquitoes engorging. A large increase in the LC90 of IVM was observed for females feeding on ATSBs and TSBs with 70% sucrose relative to those feeding on baits with lower sugar concentrations. No such effect was observed for males.

Field and Laboratory Testing of Feeding Stimulants to Enhance Insecticide Efficacy Against Spotted-Wing Drosophila, Drosophila suzukii (Matsumura).

The invasive spotted-wing drosophila, Drosophila suzukii (Matsumura), is a key insect pest of berries globally, causing lost revenues and increased production costs associated with applications of insecticides. The insecticides utilized are commonly broad-spectrum pyrethroids, organophosphates, or carbamates in conventionally managed fields and spinosad in organically managed fields. Adoption of more selective insecticides has been limited due to their lower residual activity, and the requirement that some must be ingested to be effective. We investigated the use of feeding stimulants for D. suzukii as a method to improve longevity and efficacy in a range of insecticides. In laboratory bioassays, sugar increased the efficacy of all chemical classes tested; however, the inclusion of yeast only showed a benefit with malathion. Feeding stimulants had a limited effect in some cases under field conditions. Similarly, infestation in field plots and a semifield bioassay showed no significant decreases in infestation with the inclusion of feeding stimulants for the insecticides tested in these trials. We discuss the implications of these findings for managing D. suzukii in fruit crops to help ensure the harvest of marketable fruit.

Effects of chitosan and erythritol on labellar taste neuron activity, proboscis extension reflex, daily food intake, and mortality of male and female spotted-winged drosophila, Drosophila suzukii.

In recent years, there has been interest in reduced-risk materials with insecticidal properties for the invasive pest spotted-wing drosophila, Drosophila suzukii. Here, we compared the peripheral sensitivity (via the tip-recording technique, used to monitor the neural activity of gustatory receptor neurons [GRNs]) and palatability (via the Proboscis Extension Reflex [PER]) of chitosan, a polysaccharide derived from chitin, with that of erythritol, a sugar alcohol, to male and female D. suzukii. Because in some insect species it has previously been shown that chitosan has some insecticidal properties, then treatment effects on mortality rates of male and female D. suzukii were quantified. Physiological recordings from the l-type labellar sensilla showed that erythritol evoked responses from one GRN, while chitosan elicited spiked activity from a second one. The first PER bioassay revealed that the level of response to erythritol increased significantly for males and females as the concentrations increased, and the effect of fly sex was non-significant. The second PER bioassay compared the male and female response to chitosan and erythritol each at 0.125, 0.25, 0.5, 1, and 2% concentrations. The overall female PER to erythritol was significantly greater than that exhibited by males, and no differences were noted between sexes when chitosan was evaluated. These results indicate that chitosan alone can elicit PER responses in adult D. suzukii. In the third experiment, chitosan was toxic to D. suzukii. When combined with sucrose (2%), chitosan elicited high levels (80-100%) of mortality of adult D. suzukii within 3 days, particularly in males. The presence of erythritol did not seem to increase the toxic effect of chitosan.

A Multimodal Attract-and-Kill Device for the Asian Citrus Psyllid Diaphorina citri (Hemiptera: Liviidae).

Phytophagous insects, including Asian citrus psyllids (Diaphorina citri Kuwayama), use multiple sensory modalities (vision, olfaction, and gustation,) to locate and accept host plants. We explored incorporation of several sensory cues into a multi-modal attract-and-kill device (AK device) using a three-dimensional shape to increase visibility, as well as elements of color, attractant, phagostimulant, UV reflectant, and toxicant. Attraction of adult D. citri to the device was mediated by a combination of a highly reflective yellow cylinder, a UV reflectant compound (magnesium oxide), and an odorant blend as a short-range attractant. The device surface was coated with a slow-release wax matrix (SPLAT™) augmented with a phagostimulant consisting of a 3-component blend (formic acid, acetic acid, and para-cymene) and an insecticide (ß-cyfluthrin). Psyllids landing on the device attempted to feed from the wax matrix, became intoxicated, died, and fell from the device. The device remained fully active over a period of 12 weeks partly because dead psyllids or nontargets did not adhere to the surface as occurs on adhesive yellow sticky cards, the industry standard. Laboratory and field assays showed that the device attracted and killed significantly more adult D. citri than ordinary yellow sticky cards. This device or a future iteration based on the design elements of this device is expected to contribute to sustainable and environmentally appropriate management of D. citri by exploiting the psyllid's innate behavioral responses to visual, olfactory, and gustatory stimuli.