Christopher T. Werle

A four-component synthetic attractant for Drosophila suzukii (Diptera: Drosophilidae) isolated from fermented bait headspace

BACKGROUND A mixture of wine and vinegar is more attractive than wine or vinegar to spotted wing drosophila, Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), and ethanol and acetic acid are considered key to that attractiveness. In addition to ethanol and acetic acid, 13 other wine and vinegar volatiles are antennally active to D. suzukii and might be involved in food finding. RESULTS Out of the 13 antennally active chemicals, acetoin, ethyl lactate and methionol increased fly response to a mixture of acetic acid and ethanol in field trapping experiments. A five-component blend of acetic acid, ethanol, acetoin, ethyl lactate and methionol was as attractive as the starting mixture of wine and vinegar in field tests conducted in the states of Oregon and Mississippi. Subtracting ethyl lactate from the five-component blend did not reduce the captures of flies in the trap. However, subtracting any other compound from the blend significantly reduced the numbers of flies captured. CONCLUSION These results indicate that acetic acid, ethanol, acetoin and methionol are key olfactory cues for D. suzukii when attracted to wine and vinegar, which may be food-finding behavior leading flies to fermenting fruit in nature. It is anticipated that this four-component blend can be used as a highly attractive chemical lure for detection and management of D. suzukii. Published 2013. This article is a U.S. Government work and is in the public domain in the USA.

Polistes spp. (Hymenoptera: Vespidae) orientation to wine and vinegar

Abstract Attractants are sought for trapping of Polistes spp. paper wasps when they are pestiferous. The serendipitous capture of Polistes metricus Say and Polistes bellicosus Cresson in traps baited with a wine/vinegar mixture for spotted wing drosophila, Drosophila suzukii (Matsumura) prompted experiments to determine the nature of the wasp response. Both wasp species were captured in subsequent field tests in traps baited with the same mixture of wine plus vinegar, and not in unbaited traps. Polistes bellicosus responses to wine, vinegar, ethanol (as a major volatile of wine), and acetic acid (as a major volatile of vinegar) were evaluated using a Y-tube olfactometer. In the olfactometer, P. bellicosus wasps were attracted to wine and not to vinegar. They also preferred wine alone to wine with vinegar, and were attracted to ethanol. Female wasps were deterred by acetic acid. In field tests comparing traps baited with wine, vinegar and a combination of the 2 materials, P. bellicosus and Polistes fuscatus (Fab.) were captured in traps baited with wine, but were not trapped with vinegar. The inclusion of vinegar with wine did not improve bait attractiveness in the field. We conclude that the paper wasp response to the D. suzukii bait of wine plus vinegar was largely the result of the wasp response to wine. This work constitutes the first demonstration of an attractive bait that can be used to trap P. bellicosus and P. metricus in situations where they are pestiferous, and suggests a potential source of a chemical attractant based on wine volatiles.

Ingestible insecticides for spotted wing Drosophila control: a polyol, Erythritol, and an insect growth regulator, Lufenuron

Bioassays tested insecticidal activity of Erythritol from the nutritive sweetener, Truvia, and an insect growth regulator, Lufenuron, against life stages (eggs, larvae, pupae, adults) of Drosophila melanogaster (Meigen) and Drosophila suzukii (Matsumura), the spotted wing Drosophila (SWD). These compounds were chosen for their demonstrated acute toxicity to adult and larval Drosophila and potential use on organic fruit farms. D. melanogaster fed on standard Drosophila diet media moistened with water containing known concentrations of Erythritol. Likewise, SWD consumed standard diet media as well as thawed host fruit (blackberries and blueberries) treated with solutions of Erythritol, Lufenuron or both. During the first bioassay, Erythritol at lower concentrations between 0 and 500 mm (~61 000 ppm) in water and mixed with instant diet media increased adult survival from ~80% to 97% for D. melanogaster and SWD. However, from aqueous concentrations ranging from 1750 (~414 000 ppm) to 2000 mm (~244 000 ppm), Erythritol killed 100% of adult Drosophila in culture vials. One hundred per cent mortality for SWD and D. melanogaster occurred at ≥0.5 m (~61 000 ppm) Erythritol added to diet media or topically applied to host fruit. In a second bioassay, 0.013–1.000 ppm of aqueous Lufenuron, a chitin synthase inhibitor, when added to dry diet media prevented 90–99% of SWD from reaching the pupal stage. In another assay, ~67% of SWD eggs or neonates (early first instars) died inside blackberries pre‐treated with (dipped in) a soapy solution of 10 ppm Lufenuron. Pre‐treating blackberry fruit with an Erythritol–Lufenuron mixture reduced SWD brood survival by 99%. Likewise, during our last fruit‐based bioassay, 98% of eggs and neonates died inside blueberries similarly pre‐treated. During the last experiment, Lufenuron in diet media also rendered adult females sterile. Sterility, however, dissipated over 7 days once females began feeding on a Lufenuron‐free diet media.

From a non‐target to a target: identification of a fermentation volatile blend attractive to Zaprionus indianus

The African fig fly, Zaprionus indianus Gupta, is rapidly spreading through the New World and is a new potential pest for numerous fruit crops. Methods are needed to detect and monitor Z. indianus. A recent study shows that Z. indianus can be attracted with a mixture of wine and vinegar, but there are no chemical attractants yet identified. This fly was captured incidentally as a non‐target insect in experiments to develop chemical lures, based on wine and vinegar fermentation volatiles, for Drosophila suzukii Matsumura and Drosophila melanogaster Meigen. We then generated testable hypotheses on what combination of these volatiles was involved in Z. indianus attraction to wine and vinegar. We determined through a series of trapping experiments that the blend of ethanol, acetic acid, acetoin, isoamyl acetate, methionol and ethyl hexanoate constitutes a strong attractant for Z. indianus and accounts for its attraction to the combination of wine and vinegar. These results and findings provide the first opportunity to develop a long‐lasting and consistent chemical lure for trapping of Z. indianus. Such a lure in a suitable trap should provide a good means to document the spread of the fly and determine its seasonality and abundance in new areas and crops.

Erythritol and Lufenuron Detrimentally Alter Age Structure of Wild Drosophila suzukii (Diptera: Drosophilidae) Populations in Blueberry and Blackberry

Abstract We report on the efficacy of 0.5 M (61,000 ppm) erythritol (E) in Truvia Baking Blend, 10 ppm lufenuron (L), and their combination (LE) to reduce egg and larval densities of wild populations of Drosophila suzukii (Matsumura) infesting fields of rabbiteye blueberries (Vaccinium virgatum) and blackberries (Rubus sp.). Formulations included the active ingredients (lufenuron, erythritol, or both), sugar (in control and erythritol treatments), and Dawn hand-soap applied to plants with pressurized 3-gallon garden spray tanks. The three chemical treatments (E, L, and LE) had no effect on D. suzukii ovipositing in blackberry and blueberry fruit, but they did reduce larval infestation by 75%, particularly densities of first and second instars. Erythritol and lufenuron were equally efficacious compounds as a D. suzukii ovicide and larvicide, but they did not display additive or synergistic activity. Extremely high larval mortality in control fruits show an age structure heavily skewed toward egg output.

Seasonal and spatial dispersal patterns of select ambrosia beetles (Coleoptera: Curculionidae) from forest habitats into production nurseries

Abstract Exotic ambrosia beetles (Coleoptera: Curculionidae) are important pests of ornamental tree nurseries. Although these beetles reportedly disperse in early spring from peripheral forested areas into nurseries, few studies have determined how far they fly to infest new host trees, or whether a masstrapping strategy can adequately protect a nursery crop. Field monitoring with ethanol baits in South Carolina (2011–2012), Mississippi (2013–2014), and Louisiana (2013–2014), USA, determined the timing of peak ambrosia beetle flights, dispersal distance, and optimal trap location. In addition to the well-documented spring flight peak, southeastern nursery managers may need to be aware of a second, late-summer flight. Captures from traps placed in a nursery at various distances (−25 to 200 m) from the forest—nursery interface showed a significant linear and quadratic trend in decreasing numbers of beetles captured with increasing distance from the forest in South Carolina, whereas significant linear, quadratic, and cubic trends were detected in Louisiana and Mississippi. Although captures at the nursery edge were lower than within the forest, traps placed at the nursery edge may still represent the optimal tool for both monitoring and mass-trapping programs because of easier access for personnel. Susceptible tree cultivars may gain added protection when placed deeper within nursery interiors and when baited traps line adjacent nursery edges.

Movement of Xylosandrus germanus (Coleoptera: Curculionidae) in Ornamental Nurseries and Surrounding Habitats

ABSTRACT Some exotic ambrosia beetles are damaging pests in ornamental nurseries. Xylosandrus germanus (Blandford) is the most problematic ambrosia beetle in Ohio nurseries. Movement of X. germanus in nurseries has not been characterized, and knowledge is lacking on whether infestations originate from within nurseries or surrounding habitats. Flight activity of X. germanus was monitored in nurseries and adjacent wooded areas to determine the source of beetles infesting nurseries, and characterize their movement within nurseries. Ethanol-baited bottle traps were positioned within wooded areas adjacent to commercial nurseries and within nurseries at various distances from the nursery woodlot interface. Flight activity of overwintered X. germanus occurred in wooded areas adjacent to nurseries before occurrence within nurseries. There was a direct relationship between degree-days and the distance from woodlots when X. germanus were first found in traps in spring, with earlier captures closest to wooded areas and latest ones furthest away into the nursery. X. germanus appeared to move into nurseries from adjacent wooded areas, with numbers trapped within nurseries decreasing with distance away from wooded areas. Trees in the interior of nurseries would appear to be subjected to less attack pressure than trees near the nursery border. Intercepting beetles as they move into nurseries might be an effective strategy to reduce attack pressure on valuable trees.

Ambrosia Beetle (Coleoptera: Curculionidae: Scolytinae) Captures Using Colored Traps in Southeast Tennessee and South Mississippi

Abstract Ambrosia beetles (Coleoptera: Curculionidae: Scolytinae) have become a major impediment to profitable nursery production, with proper controls centered around an accurate monitoring program. Whereas the use of semiochemicals by ambrosia beetles is well understood, the potential use of visual cues including colors remains relatively understudied. Field tests were performed in Tennessee and Mississippi to determine response of the ambrosia beetle community to 13 trap colors (black, blue, brown, clear, gray, green, lavender, opaque, orange, purple, red, white, and yellow). Traps consisted of corrugated plastic in the shape of a prism and covered in insect glue (Pestick™ or Tangle-Trap®). All trap colors were analyzed with a spectro-photometer to quantify spectral reflectance, ranging from 6 - 80% reflectance. Mean ambrosia beetle capture from opaque and red traps (60 and 54, respectively) was significantly higher than from yellow or white traps (30 and 28, respectively). However, captures from each of the afore-mentioned 4 colors were not significantly different from the other 9 colors tested. Although further research is needed to explain the efficacy of the opaque traps, we can recommend that industry-standard black traps are an acceptable choice for ambrosia beetle monitoring programs; whereas, yellow or white traps are not recommended.

Laboratory and Field Assessments of Erythritol Derivatives on the Survival, Reproductive Rate, and Control of Drosophila suzukii (Diptera: Drosophilidae)

Abstract Spotted-wing Drosophila, Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) is a vinegar fly introduced unintentionally into the United States. Since 2008, D. suzukii has reduced annual berry yields from 6 to 100%. Effective control of D. suzukii during harvest requires weekly applications of low-residual, broad-spectrum insecticides that are unavailable for organic farming. A novel ingestible insecticide, a 4-carbon polyol, mesoerythritol (erythritol), was found to kill 75 to 100% of larval and adult D. suzukii. However, mesoerythritol, at effective concentrations (0.5–1.0M), may be cost-prohibitive. Therefore, we conducted laboratory tests to assess the effects of lower cost derivatives of erythritol, namely the pentaerythritol series of 1,3-diols on D. suzukii pupal production, adult production, adult mortality, brood output, and reproductive increase. We then selected the two most promising compounds for a field test on fruiting rabbiteye blueberry. From 90 to 100% of adults died when fed food moistened with 1M solutions of mesoerythritol and pentaerythritol. Mesoerythritol and dipentaerythritol at a concentration of 1M were ovicidal/ larvicidal, killing ≥85% of immatures. Overall, 1M mesoerythritol killed 80% or more larvae and adults, thus bringing populations to near zero. The heaviest compound of this series, tripentaerythritol, at all concentrations, was largely benign to both adults and immatures. Thus, we cannot recommend tripentaerythritol for D. suzukii control. In a blueberry field, 0.5M mesoerythritol and 0.5M pentaerythritol, each by themselves, reduced egg infestation by 64% and larval infestation by 93%; their combination (0.25M mesoerythritol and 0.25M pentaerythritol) achieved even greater egg control with 82% fewer eggs infesting blueberry fruits.

Development of Microsatellites for Population Genetic Analyses of the Granulate Ambrosia Beetle (Coleoptera: Curculionidae)

Limited male dispersal and local mating in ambrosia beetles are expected to result in extreme inbreeding and highly structured populations. In this study, we developed microsatellite markers for the granulate ambrosia beetle, Xylosandrus crassiusculus (Motschulsky), for use in future studies into population and family structure of this invasive pest species. We employed de novo next-generation sequencing to generate whole genome shotgun sequences for the characterization of microsatellite loci. Approximately 6% of the 84,024 contigs generated from Hi-Seq Illumina 2x250bp sequencing contained microsatellites with at least four repeats of di-, tri-, tetra-, penta-, and hexamers. Primers were synthesized for 40 microsatellite loci with trimer repeat units. Twenty-four primer pairs yielded consistent PCR products of unique loci and were validated for population genetic application using three sample groups each containing 20 X. crassiusculus individuals from Mississippi. Thirteen loci were found to be polymorphic with up to five alleles per population. The two beetle sample groups from Pearl River County (Poplarville and McNeill) belonged genetically to the same population. The population from Lamar County (Purvis) was genetically distinct, separated by a moderate genetic distance (FST = 0.11) and five unique alleles (with >5% frequency). Consistent with the perceived mating structure (incest of females with flightless males), the populations showed homozygote excess at most loci, as indicated by the coefficients of inbreeding (FIT = 0.45 and FIS = 0.37) and high mean relatedness among individuals (r = 0.15).