Comparing the effectiveness of different insecticide application orders for suppressing Drosophila suzukii Matsumura (Diptera: Drosophilidae) infestation: experimental and modeling approaches.

Drosophila suzukii Matsumura (Diptera: Drosophilidae) is a key pest of soft-skinned fruit such as blackberry and blueberry. Differing seasonal spray regimes are expected to have variable effects on D. suzukii populations. Semi-field cage trials were performed at three locations in the United States (Georgia, Oregon, and North Carolina) on blueberry and blackberry crops to evaluate this hypothesis. Insecticides with different efficacy rates (ZC - zeta-cypermethrin, SPI - spinetoram, CYAN - cyantraniliprole) were applied during field experiments conducted within large cages. Treatment schedules consisted of two insecticide applications which performed over three weeks. Seasonal treatment schedules were applied in the following order: ZC-CYAN and CYAN-ZC in rabbiteye and highbush blueberry with the addition of a ZC-SPI treatment applied in blackberry. In addition, a population model was applied to simulate the relative efficacy of the insecticide schedules in Oregon on D. suzukii population model based on previously published efficacy, biological, and weather parameters. Overall, all schedules resulted in reduced D. suzukii infestation compared to untreated control (UTC) treatments, with statistical differences in all three locations. The numerically lower infestation was found in some cases in ZC-CYAN schedule. Population modeling conducted exclusively for blueberry, and the simulations indicated no discernible differences between the two respective schedules (ZC-CYAN vs CYAN-ZC). The present study demonstrates that seasonal infestation of D. suzukii could be suppressed irrespective of application order. Additional research is required to assess the optimal timing and sequence of insecticide applications for controlling seasonal populations of D. suzukii in fruit crops. Such information could be invaluable for growers who are seeking to strategize their insecticide applications.

Open field trials of food-grade gum in California and Oregon as a behavioral control for Drosophila suzukii Matsumura (Diptera: Drosophilidae).

The invasion of Drosophila suzukii, spotted-wing drosophila, across Europe and the US has led to economic losses for berry and cherry growers, and increased insecticide applications to protect fruit from damage. Commercial production relies heavily on unsustainable use of conventional toxic insecticides. Non-toxic insecticide strategies are necessary to alleviate the disadvantages and non-target impacts of toxic conventional insecticides and improve Integrated Pest Management (IPM). A novel food-grade gum deployed on dispenser pads (GUM dispensers) was evaluated to mitigate D. suzukii crop damage in five commercial crops and nine locations. Trials were conducted at a rate of 124 dispensers per hectare in cherry, wine grape, blueberry, raspberry, and blackberry in California and Oregon, USA during 2019 and 2020. The majority of trials with the food-grade gum resulted in a reduction of D. suzukii egg laying in susceptible fruit. In some cases, such damage was reduced by up to 78%. Overall, results from our meta-analysis showed highly significant differences between GUM treatments and the untreated control. Modeling simulations suggest a synergistic reduction of D. suzukii damage when used in combination with Spinosad (Entrust SC) insecticide. These data illustrate commercial value of this tool as a sustainable alternative to manage D. suzukii populations within a systems approach.

A Horticultural Cuticle Supplement Can Impact Quality Characters and Drosophila suzukii Damage of Several Small and Stone Fruit.

Surface wax and other cellular building blocks play an important role in preserving fruit integrity from biotic and abiotic adversities. Huge energy expenses are made by plants to place these protective compounds onto the epidermal cuticle. Sprayable plant and fruit coatings have been developed to protect plant tissues from environmental stresses, pathogens, and arthropods. The aim of this study was to determine if an experimental cuticle supplement containing waxes can affect fruit quality parameters such as firmness and size of various crops. Cherry, blueberry, and winegrape plants treated with the cuticle supplement showed significant increases in berry firmness ranging from 4.6 to 11.6%. No quality benefits were however observed on blackberry. Cuticle supplement applications did not significantly affect berry size. Laboratory trials resulted in a 54% mean reduction in a model pest insect i.e., Drosophila suzukii egg laying on blueberry. Short-duration field trials over 72 ±â€…2 to 96 ±â€…2 h on commercial-standard blueberry bushes resulted in 50-93.4% reductions of D. suzukii damage. Longer-term field trials on cherry and blueberry challenged with egg-laying D. suzukii showed reductions of damage ranging from 45 to 95%, up to 30 d after initial cuticle supplement applications. These results indicate that the cuticle supplement significantly alters berry firmness and reduces D. suzukii damage under commercial production conditions. One factor that may contribute to this reduction includes improved fruit quality parameters. The current work serves to expand integrated pest management options to control D. suzukii populations in commercial field settings.

Comparative Insecticide Application Techniques (Micro-Sprinkler) Against Drosophila suzukii Matsumura (Diptera: Drosophilidae) in Highbush Blueberry.

Drosophila suzukii (Matsumura), spotted-wing drosophila, is a major pest in small fruit crops including highbush blueberry. Controlling D. suzukii is challenging and chemical control is the main method to manage D. suzukii populations. Growers have expressed interest in using micro-sprinklers as an alternative method to apply insecticides. The current study aimed to evaluate if insecticide applications using micro-sprinklers can be used as an alternative method to protect the fruit from D. suzukii egg-laying. Modeling was used as an additional tool to parameterize the relative insecticide efficacy on oviposition. Field measurements of different treatments were conducted over periods of eleven days on commercial-standard highbush blueberry. Cyantraniliprole and spinetoram were applied using both a micro-sprinkler and a backpack sprayer. Treatments of Chromobacterium subtsugae and zeta-cypermethrin were only applied using a backpack sprayer. Both cyantraniliprole and spinetoram treatments resulted in moderate suppression of D. suzukii egg-laying. No statistical significance was found between micro-sprinkler and backpack sprayer applications for these two insecticides. Zeta-cypermethrin treatments using a backpack sprayer resulted in the most significant suppression of D. suzukii egg-laying over eleven days, while C. subtsugae was less effective at preventing D. suzukii egg-laying. Modeling simulations estimate the impact of the control methods on D. suzukii populations dynamics and simulation outputs indicated that backpack sprayers reduced D. suzukii populations at more pronounced levels compared to micro-sprinkler applications. The present study indicates that there is an underlying value of micro-sprinkler systems as an alternative and rapid spray application technique to help suppress D. suzukii pest populations during high-pressure periods in highbush blueberry production.

Drosophila suzukii (Diptera: Drosophilidae): A Decade of Research Towards a Sustainable Integrated Pest Management Program.

Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) also known as spotted-wing drosophila (SWD), is a pest native to Southeast Asia. In the last few decades, the pest has expanded its range to affect all major European and American fruit production regions. SWD is a highly adaptive insect that is able to disperse, survive, and flourish under a range of environmental conditions. Infestation by SWD generates both direct and indirect economic impacts through yield losses, shorter shelf life of infested fruit, and increased production costs. Fresh markets, frozen berries, and fruit export programs have been impacted by the pest due to zero tolerance for fruit infestation. As SWD control programs rely heavily on insecticides, exceedance of maximum residue levels (MRLs) has also resulted in crop rejections. The economic impact of SWD has been particularly severe for organic operations, mainly due to the limited availability of effective insecticides. Integrated pest management (IPM) of SWD could significantly reduce chemical inputs but would require substantial changes to horticultural management practices. This review evaluates the most promising methods studied as part of an IPM strategy against SWD across the world. For each of the considered techniques, the effectiveness, impact, sustainability, and stage of development are discussed.

Electrophysiological Responses of the Mediterranean Fruit Fly, Ceratitis capitata, to the Cera Trap® Lure: Exploring Released Antennally-Active Compounds.

The Mediterranean fruit fly (medfly), Ceratitis capitata, is a worldwide pest of agriculture able to use olfactory cues to locate habitat, food sources, mates and oviposition sites. The sensitivity of medfly olfaction has been exploited to develop olfactory-based attractants that are currently important tools for detection, control and eradication of its populations. Among these is Cera Trap® (BIOIBERICA, S.A.U.), a cost-effective bait. Here we used coupled gas chromatography/electroantennographic detection (GC-EAD) and GC/mass spectrometry (GC-MS) approaches to characterize the medfly antennally-active compounds released by this lure. We identified GC peaks corresponding to chemicals belonging to six different classes including heterocyclic aromatic compounds, aliphatic alcohols, aldehydes, esters, sesquiterpene hydrocarbons, and aromatic alcohols. We tested ten potential candidate volatiles belonging to these classes and predicted to be emitted by the lure and found that they were eliciting electroantennographic responses in medfly adults. These results will help in unravelling the physiological mechanisms of odor perception in both sexes, especially in relation to Cera Trap® attractant activity, which in the field has been shown to be female-specific. These findings and their developments will ultimately expand the toolbox for medfly control in the field.

Structural and transcriptional evidence of mechanotransduction in the Drosophila suzukii ovipositor.

Drosophila suzukii is an invasive pest that prefers to lay eggs in ripening fruits, whereas most closely related Drosophila species exclusively use rotten fruit as oviposition site. This behaviour is allowed by an enlarged and serrated ovipositor that can pierce intact fruit skin, and by multiple contact sensory systems (mechanosensation and taste) that detect the optimal egg-laying substrates. Here, we tested the hypothesis that bristles present in the D. suzukii ovipositor tip contribute to these sensory modalities. Analysis of the bristle ultrastructure revealed that four different types of cuticular elements (conical pegs type 1 and 2, chaetic and trichoid sensilla) are present on the tip of each ovipositor plate. All of them have a poreless shaft and are innervated at their base by a single neuron that ends in a distal tubular body, thus resembling mechanosensitive structures. Fluorescent labelling in D. suzukii and D. melanogaster revealed that pegs located on the ventral side of the ovipositor tip are innervated by a single neuron in both species. RNA-sequencing profiled gene expression, notably sensory receptor genes of the terminalia of D. suzukii and of three other Drosophila species with changes in their ovipositor structure (from serrated to blunt ovipositor: Drosophila subpulchrella, Drosophila biarmipes and D. melanogaster). Our results revealed few species-specific transcripts and an overlapping expression of candidate mechanosensitive genes as well as the presence of some chemoreceptor transcripts. These experimental evidences suggest a mechanosensitive function for the D. suzukii ovipositor, which might be crucial across Drosophila species independently from ovipositor shape.

Reproductive Site Selection: Evidence of an Oviposition Cue in a Highly Adaptive Dipteran, Drosophila suzukii (Diptera: Drosophilidae).

Drosophila suzukii (Matsumura) is a vinegar fly species that originates from Eastern Asia and has spread throughout Europe and the Americas since its initial detection in United States in 2008. Its relatively large, sclerotized, and serrated ovipositor enables the ability to penetrate ripening fruits, providing a protected environment for its egg and larval stages. Because the mechanism of oviposition site selection of D. suzukii is a matter of hypothesis, the aim of the present study was to elucidate behavioral and chemical aspects of short-range ovipositional site selection within the context of D. suzukii reproductive biology. The preference of D. suzukii to lay eggs on artificially pierced, previously infested, or intact fruits was tested. Video recordings and photographic evidence documented the release of an anal secretion over the fruit surface near the oviposition sites. Gas chromatographic analysis revealed the presence of 11 compounds detected only on the skin of egg-infested berries. Electroantennographic experiments with both sexes of D. suzukii highlighted the importance of six volatile compounds: methyl myristate, methyl palmitate, myristic acid, lauric acid, palmitic acid, and palmitoleic acid. Finally, a synthetic blend composed of the six compounds in a ratio similar to that found on the skin of egg-infested berries increased the oviposition rate of conspecific females. Data from our work suggest that the identified volatiles are cues for reproductive site selection. We discuss how these oviposition cues may affect the fitness of D. suzukii. The knowledge gained from this study may accelerate establishment of control strategies based on the interference and disruption of D. suzukii communication during the oviposition processes.

Interactions among morphotype, nutrition, and temperature impact fitness of an invasive fly.

Invasive animals depend on finding a balanced nutritional intake to colonize, survive, and reproduce in new environments. This can be especially challenging during situations of fluctuating cold temperatures and food scarcity, but phenotypic plasticity may offer an adaptive advantage during these periods. We examined how lifespan, fecundity, pre-oviposition periods, and body nutrient contents were affected by dietary protein and carbohydrate (P:C) ratios at variable low temperatures in two morphs (winter morphs WM and summer morphs SM) of an invasive fly, Drosophila suzukii. The experimental conditions simulated early spring after overwintering and autumn, crucial periods for survival. At lower temperatures, post-overwintering WM lived longer on carbohydrate-only diets and had higher fecundity on low-protein diets, but there was no difference in lifespan or fecundity among diets for SM. As temperatures increased, low-protein diets resulted in higher fecundity without compromising lifespan, while high-protein diets reduced lifespan and fecundity for both WM and SM. Both SM and WM receiving high-protein diets had lower sugar, lipid, and glycogen (but similar protein) body contents compared to flies receiving low-protein and carbohydrate-only diets. This suggests that flies spend energy excreting excess dietary protein, thereby affecting lifespan and fecundity. Despite having to recover from nutrient depletion after an overwintering period, WM exhibited longer lifespan and higher fecundity than SM in favorable diets and temperatures. WM exposed to favorable low-protein diet had higher body sugar, lipid, and protein body contents than SM, which is possibly linked to better performance. Although protein is essential for oogenesis, WM and SM flies receiving low-protein diets did not have shorter pre-oviposition periods compared to flies on carbohydrate-only diets. Finding adequate carbohydrate sources to compensate protein intake is essential for the successful persistence of D. suzukii WM and SM populations during suboptimal temperatures.

Functional transcriptome analyses of Drosophila suzukii antennae reveal mating-dependent olfaction plasticity in females.

Insect olfaction modulates basal behaviors and it is often influenced by the physiological condition of each individual such as the reproductive state. Olfactory plasticity can be achieved by modifications at both peripheral and central nervous system levels. Here we performed a genome-wide transcriptomic analysis of the main olfactory organ, the antenna, to investigate how gene expression varies with female mating status in Drosophila suzukii, a destructive and invasive soft fruit pest. We observed a wide mating-induced up-regulation of chemosensory-related genes in females, especially odorant receptor (Or) genes. We then used a candidate gene approach to define the comprehensive dataset of antenna-expressed chemosensory receptors and binding proteins, which showed many similarities with Drosophila melanogaster. Candidate gene approach was also used to finely quantify differential expression at Or isoform level, suggesting post-mating transcriptional modulation of genes involved in the peripheral olfactory system. We identified 27 up-regulated Or transcripts encoded by 25 genes, seven of them were duplications specific to D. suzukii lineage. Post-mating olfactory modulation was further supported by electroantennogram recordings that showed a differential response according to mating status to one out of eight odors tested (isoamyl-acetate). Our study characterizes the transcriptional mechanisms driven by mating in D. suzukii female antennae. Understanding the role of genes differentially expressed in virgin or mated females will be crucial to better understand host finding and the crop-damaging oviposition behavior of this species.

Survival and Fecundity Parameters of Two Drosophila suzukii (Diptera: Drosophilidae) Morphs on Variable Diet Under Suboptimal Temperatures.

Life history parameters are used to estimate population dynamics, mortality, and reproduction in insects relative to their surrounding environment. For Drosophila suzukii Matsumura (Diptera: Drosophilidae), an invasive agricultural pest, previous studies have estimated net reproductive rate (Ro), generation time (T), and intrinsic rate of population increase (rm). A main limitation is that these estimates were measured under relatively favorable settings, and do not reflect environmental conditions and physiological states encountered during dormancy periods. Therefore, this study investigated the impacts of 1) low temperatures and 2) dietary protein: carbohydrate ratios (P:C) on both survival and fecundity parameters of D. suzukii summer morphs (SM) and postoverwintering winter morphs (WM) over physiological age (degree-days, DD). In both morphs, reproductive rates were higher and lifespan was longer when flies were exposed to low protein (P:C 1:4) or carbohydrate-only diets (P:C 0:1) compared with high protein diets (P:C 1:1). WM had higher reproductive rates and longer generation times than SM on optimal 1:4 diet in all trialed temperatures, but at the lowest temperatures, SM had higher reproductive rates than WM in carbohydrate-only and high protein diets. This likely reflected delayed oogenesis and hindered reproduction after an overwintering period in WM receiving suboptimal diets. Oviposition for SM and WM receiving 1:4 diet commenced from 0 to 100 DD, and peaked between 400 and 500 DD, earlier than flies receiving 0:1 diet. These results suggest that dietary protein has a crucial role in early oogenesis, particularly for postoverwintering WM. The parameters developed here reflect the population dynamics of D. suzukii before and after the crop growing season, an essential time for population buildup, survival, and early and late host infestation.

Seasonal Reproductive Biology of Drosophila suzukii (Diptera: Drosophilidae) in Temperate Climates.

Drosophila suzukii (Matsumura; Diptera: Drosophilidae) is a key pest of sweet cherry and small fruits worldwide. The present studies were designed to describe the reproductive physiology in both sexes, through dissections of their reproductive organs. We extensively dissected female D. suzukii throughout the season from 2013 to 2016 and classified the reproductive status flies based on five recognizable ovarian maturation stages: 1) no ovaries; 2) unripe ovaries 3) ripening eggs in ovarioles; 4) mature eggs in ovarioles; and 5) mature eggs in the abdomen. Development was examined as a function of calendar days as well as degree-days (DD). Results obtained from winter collections revealed that females collected from November to March contained a lower percentage of mature eggs than females collected from April to September. These data suggest that environmental conditions during the dormant period induce reproductive diapause. Oogenesis likely increased with an increase in mean monthly temperatures and DD. The first overwintered females with mature eggs were dissected as early as 21 February 2014 in Trento (7 DD). Additionally, we found that a low proportion of males (less than 50%) had sperm in their testes between January and March, yet during the same period females already have sperm stored in their spermathecal. Ivy berries was an alternative but unfavorable non-crop host during the late dormant period, as evidenced by emergence of smaller adults when compared to individuals emerging from cherry fruits. This study showed that D. suzukii females have great potential for oviposition early in the season, posing a risk to early season maturing crop hosts.

Genetic variability in Italian populations of Drosophila suzukii.

BACKGROUND: Drosophila suzukii is a highly destructive pest species, causing substantial economic losses in soft fruit production. To better understand migration patterns, gene flow and adaptation in invaded regions, we studied the genetic structure of D. suzukii collected across Italy, where it was first observed in 2008. In particular, we analysed 15 previously characterised Simple Sequence Repeat (SSR) markers to estimate genetic differentiation across the genome of 278 flies collected from nine populations. RESULTS: The nine populations showed high allelic diversity, mainly due to very high heterozygosity. The high Polymorphism Information Content (PIC) index values (ranging from 0.68 to 0.84) indicated good discrimination power for the markers. Negative fixation index (F IS) values in seven of the populations indicated a low level of inbreeding, as suggested by the high number of alleles. STRUCTURE, Principal Coordinate and Neighbour Joining analysis also revealed that the Sicilian population was fairly divergent compared to other Italian populations. Moreover, migration was present across all populations, with the exception of the Sicilian one, confirming its isolation relative to the mainland. CONCLUSIONS: This is the first study characterising the genetic structure of the invasive species D. suzukii in Italy. Our analysis showed extensive genetic homogeneity among D. suzukii collected in Italy. The relatively isolated Sicilian population suggests a largely human-mediated migration pattern, while the warm climate in this region allows the production of soft fruit, and the associated D. suzukii reproductive season occurring much earlier than on the rest of the peninsula.

Molecular markers for analyses of intraspecific genetic diversity in the Asian Tiger mosquito, Aedes albopictus.

BACKGROUND: The dramatic worldwide expansion of Aedes albopictus (the Asian tiger mosquito) and its vector competence for numerous arboviruses represent a growing threat to public health security. Molecular markers are crucially needed for tracking the rapid spread of this mosquito and to obtain a deeper knowledge of population structure. This is a fundamental requirement for the development of strict monitoring protocols and for the improvement of sustainable control measures. METHODS: Wild population samples from putative source areas and from newly colonised regions were analysed for variability at the ribosomal DNA internal transcribed spacer 2 (ITS2). Moreover, a new set of 23 microsatellite markers (SSR) was developed. Sixteen of these SSRs were tested in an ancestral (Thailand) and two adventive Italian populations. RESULTS: Seventy-six ITS2 sequences representing 52 unique haplotypes were identified, and AMOVA indicated that most of their variation occurred within individuals (74.36%), while only about 8% was detected among populations. Spatial analyses of molecular variance revealed that haplotype genetic similarity was not related to the geographic proximity of populations and the haplotype phylogeny clearly indicated that highly related sequences were distributed across populations from different geographical regions. The SSR markers displayed a high level of polymorphism both in the ancestral and in adventive populations, and F ST estimates suggested the absence of great differentiation. The ancestral nature of the Thai population was corroborated by its higher level of variability. CONCLUSIONS: The two types of genetic markers here implemented revealed the distribution of genetic diversity within and between populations and provide clues on the dispersion dynamics of this species. It appears that the diffusion of this mosquito does not conform to a progressive expansion from the native Asian source area, but to a relatively recent and chaotic propagule distribution mediated by human activities. Under this scenario, multiple introductions and admixture events probably play an important role in maintaining the genetic diversity and in avoiding bottleneck effects. The polymorphic SSR markers here implemented will provide an important tool for reconstructing the routes of invasion followed by this mosquito.