Fumagilin-B is used to treat nosema infection in honey bee colonies; however, it is unclear whether treatment consistently reduces Vairimorpha ceranae (Fries et al.) abundance and improves colony strength and survival in the Canadian Prairies. This study assessed spring and fall fumagillin treatments on nosema abundance, colony strength, and mortality in 2 different beekeeping regions within Alberta, using both indoor and outdoor wintering management at each site. We compared 4 fumagillin treatments: Spring-only, Fall-only, Spring-and-Fall, and Control (no treatment). The spring treatment dose was ∼68 mg/colony, whereas the fall treatment dose was 120 or 48 mg/colony, depending on the year. We found that the colonies were infected predominately with V. ceranae, with V. apis (Zander) present only in mixed infections in a subset of colonies. Although treatment in either the spring or fall did reduce nosema abundance in the short term, it did not eliminate the infection, making continued monitoring necessary. Colony strength was improved by spring treatment in some locations but not consistently, possibly due to the treatment timing or low dose. The combined spring and fall treatment increased colony survival over winter in one of 2 yr. Wintering method did not interact with treatment to affect nosema abundance in the spring. There does not appear to be a significant residual benefit of fall treatment as it did not reduce spring nosema abundance or increase colony population. Therefore, spring treatment should be applied to reduce spring V. ceranae abundance rather than relying on residual efficacy from previous fall treatments.

Pollen is an essential component of bee diets, and rearing bumble bees (Bombus spp.) for commercial use necessitates feeding pollen in mass quantities. This pollen is collected from honey bee (Apis mellifera L.) colonies because neither an artificial diet nor an economical, large-scale pollen collection process from flowers is available. The provenance of honey bee-collected pollen is often unknown, and in some cases has crossed international borders. Both deformed wing virus (DWV) and the fungal pathogen Ascosphaera apis (Claussen) Olive & Spiltoir (cause of chalkbrood disease); occur in honey bee-collected pollen, and infections have been observed in bumble bees. We used these pathogens as general surrogates for viruses and spore-forming fungal diseases to test the efficacy of 3 sterilization methods, and assessed whether treatment altered pollen quality for the bumble bee. Using honey bee-collected pollen spiked with known doses of DWV and A. apis, we compared gamma irradiation (GI), ozone fumigation (OZ), and ethylene oxide fumigation (EO) against an untreated positive control and a negative control. Following sterilization treatments, we tested A. apis spore viability, detected viral presence with PCR, and tested palatability to the bumble bee Bombus impatiens Cresson. We also measured bacterial growth from pollens treated with EO and GI. GI and EO outperformed OZ treatment in pathogen suppression. EO had the highest sterilizing properties under commercial conditions and retained palatability and supported bee development better than other treatments. These results suggest that EO sterilization reduces pathogen risks while retaining pollen quality as a food source for rearing bumble bees.

Graphical Abstract

Due to a lack of knowledge on the pollination requirements of kiwifruit cultivars grown within the United States, farmers simultaneously implement multiple pollination methods, like the rental of managed bee species or artificial pollination to achieve high fruit yields. However, implementing multiple pollination methods is costly and possibly an inefficient use of resources. We assessed the contribution of two managed bees (Apis mellifera and Bombus impatiens) to the pollination of kiwifruit by i) determining the relative abundance of kiwifruit pollen collected by foragers of each bee species, and ii) comparing fruit set and fruit quality among insect and artificially pollinated flowers through an insect exclusion experiment. A significant difference was observed between the mean relative abundance of kiwifruit pollen carried in the corbicula of A. mellifera and B. impatiens, with B. impatiens carrying on average 46% more kiwifruit pollen than A. mellifera. Artificially pollinated kiwifruit flowers set significantly greater numbers of fruit per flower at four weeks post-bloom and at harvest compared to insect pollination, wind pollination, and pollen exclusion treatment. Artificial pollination produced fruits of greater weight, size, and seed number compared to insect-pollinated flowers, and few fruits were produced in the pollen exclusion and wind pollination treatments. Kiwifruit producers experiencing similar conditions to ours should focus on artificially pollinating their crops rather than relying on managed or wild insects for kiwifruit pollination. Future research should evaluate other methods of artificial pollination to determine their effectiveness, efficiency, and economics in the pollination of kiwifruit grown within the United States.

Canadian beekeepers faced widespread levels of high honey bee colony mortality over the winter of 2021/2022, with an average winter loss of 45%. To understand the economic impact of winter colony mortality in Canada and the beekeeping management strategies used to mitigate these losses, we develop a profit model of commercial beekeeping operations in Alberta, Canada. Our model shows that for operations engaging in commercial pollination as well as honey production (compared to honey production alone), per colony profit is higher and operations are better able to withstand fluctuations in exogenous variables such as prices and environmental factors affecting productivity including winter mortality rates. The results also suggest that beekeeping operations that replace winter colony losses with splits instead of package bees accrue higher per colony profit than those importing packages to replace losses. Further, operations that produce their own queens to use in their replacement splits, accrue even higher profit. Our results demonstrate that the profitability of beekeeping operations is dependent on several factors including winter mortality rates, colony replacement strategies, and the diversification of revenue sources. Beekeepers who are not as susceptible to price and risk fluctuations in international markets and imported bee risks accrue more consistently positive profits.

Does Xylella fastidiosa, a bacterial plant pathogen with noncirculative foregut-borne transmission, manipulate behavior of its sharpshooter vector to facilitate its own inoculation? To answer this question, blue-green sharpshooters, Graphocephala atropunctata (Signoret), were reared on basil to clean their foreguts, then removed from the colony and given one of four pre-electropenetrography (EPG) treatments: i) old colony adults on basil, ii) young colony adults on basil, iii) young colony adults held on healthy grapevine for 4 days, and iv) young colony adults held on Xf-infected (symptomatic) grapevine for 4 days. After treatments, stylet probing behaviors were recorded on healthy grapevine via AC-DC electropenetrography. Waveforms representing putative Xf inoculation (XB1 [salivation and rinsing egestion] and XC1 [discharging egestion]) and other behaviors were statistically compared among treatments. Mean number of events per insect and ‘total’ duration per insect of XB1 and XC1 were highest for insects from healthy grape, lowest for basil (regardless of insect age), and intermediate for Xf-infected grape. The surprising results showed that prior exposure to healthy grapevines had a stronger effect on subsequent performance of inoculation behaviors on healthy grapevine than did prior exposure to Xf-infected grapevine. It is hypothesized that non-Xf microbes were acquired from healthy grapevine, causing greater clogging of the precibarium, leading to more performance of inoculation behaviors.This study shows for the first time that presence of noncirculative, foregut-borne microbes can directly manipulate a vector’s behavior to increase inoculation. Also, EPG can uniquely visualize the dynamic interactions between vectors and the microbes they carry.

Potato leafroll virus (PLRV) has been well managed by neonicotinoids since their widespread adoption in the United States, becoming virtually absent from seed and production fields in the Northwest. However, with increasing interest in discontinuing neonicotinoid usage, there is concern that PLRV could enjoy a resurgence in the absence of effective alternative chemistries. We tested the effects of afidopyropen, an insecticide with novel mode of action, on PLRV transmission and the feeding/probing behavior of its primary vector, the green peach aphid (Myzus persicae [Sulzer]). Afidopyropen foliar sprays decreased PLRV transmission by individual green peach aphids relative to water controls: PLRV acquisition from treated potato plants and subsequent transmission was reduced by 89%, and PLRV inoculation by viruliferous aphids to treated potato plants was reduced by 35%. Although electropenetrograph analyses following 4-h recordings of individual aphids on potato plants showed decreases in the total and mean duration of phloem feeding (E) on plants subject to an afidopyropen treatment relative to water, these only trended toward significance. Taken together, these results suggest treatment with afidopyropen can decrease PLRV transmission in potatoes, but that significant changes in feeding/probing might not occur quickly post-exposure. Overall, while the reductions in transmission were not as dramatic as have been observed following neonicotinoid treatments, afidopyropen may be a useful alternative and should be evaluated in field experiments.

Cotton leafroll dwarf virus (CLRDV) is a yield-limiting, aphid-transmitted virus that was identified in cotton, Gossypium hirsutum L., in the United States of America in 2017. CLRDV is currently classified in the genus Polerovirus, family Solemoviridae. Although 8 species of aphids (Hemiptera: Aphididae) are reported to infest cotton, Aphis gossypii Glover is the only known vector of CLRDV to this crop. Aphis gossypii transmits CLRDV in a persistent and nonpropagative manner, but acquisition and retention times have only been partially characterized in Brazil. The main objectives of this study were to characterize the acquisition access period, the inoculation access period, and retention times for a U.S. strain of CLRDV and A. gossypii population. A sub-objective was to test the vector competence of Myzus persicae Sulzer and Aphis craccivora Koch. In our study, A. gossypii apterous and alate morphs were able to acquire CLRDV in 30 min and 24 h, inoculate CLRDV in 45 and 15 min, and retain CLRDV for 15 and 23 days, respectively. Neither M. persicae nor A. craccivora acquired or transmitted CLRDV to cotton.

Bioassays involving newly hatched larval insects can be limited by the larvae's feeding state. Assays attempting to monitor mortality effects can be negatively affected by starvation effects on the larvae. Neonate western corn rootworms have significant reductions in viability if not provided food within 24 h post hatch. The recent development of an improved artificial diet for western corn rootworm larvae provides a new bioassay type for evaluating entomopathogenic nematodes that also makes the testing arena easy to observe. Here, we evaluated four species of entomopathogenic nematodes including Heterorhabditis bacteriophora Poinar, Steinernema carpocapsae (Weiser), Steinernema diaprepesi Nguyen & Duncan, and Steinernema rarum (de Doucet) against neonate western corn rootworm, Diabrotica virgifera virgifera LeConte, in 96-well plate diet bioassays. Nematode inoculation levels were 0, 15, 30, 60, and 120 nematodes per larva. Percentage mortality increased for each species as the rate of inoculation increased. Overall, H. bacteriophora and S. carpocapsae caused the greatest amount of larval mortality. The diet-based bioassays were shown to be an effective method for nematode exposure to insect pests. The assays provided adequate moisture to keep nematode from desiccating while also allowing freedom of movement around the arenas. Both rootworm larvae and nematodes were contained within the assay arenas. The addition of nematodes did not cause any significant deterioration of the diet within the three-day period of testing. Overall, the diet bioassays worked well as a measure of entomopathogenic nematode virulence.

During the past decade, the use of predatory mirids alone or combined with releases of egg parasitoids of the genus Trichogramma have been tested in Europe for biological control of the worldwide invasive pest, Tuta absoluta (Meyrick). Here, we evaluated the control of this pest by the release of the Neotropical mirid Macrolophus basicornis (Stal), the Neotropic/Nearctic parasitoid Trichogramma pretiosum Riley, and by combined releases of the predator and the parasitoid. Tests were conducted in greenhouse compartments during the summer and fall season. Each compartment contained 10 tomato plants in which only the pest was released or the pest with 1 or 2 natural enemies. Plant damage, and pest and natural enemy densities were checked weekly on one apical, medium, and bottom leaf of 5 plants. Both M. basicornis and T. pretiosum significantly reduced T. absoluta density when released alone. Combined releases resulted in a 10% higher reduction during the summer season, but not during the fall season. The damage caused by T. absoluta was significantly higher in control treatments than in all natural enemy treatments: at the end of the summer trial leaves were completely damaged in the control treatment, whereas only up to 25% leaf damage occurred in the natural enemy treatments. Combined releases did not result in lower damage than with releases of either M. basicornis or T. pretiosum. Practical aspects of single and combined releases are discussed.

Graphical Abstract

In Xinjiang's cotton growing area of China, previous studies have shown that matrine is a selective botanical insecticide, with high toxicity to Aphis gossypii Glover (Hemiptera: Aphididae) and low toxicity to its dominant natural enemy, Hippodamia variegata Goeze (Coleoptera: Coccinellidae). However, lethal effects alone are not sufficient evidence to justify introducing matrine into local IPM strategies. In this context, we systematically evaluated the safety of matrine to H. variegata by investigating the effects of contact and stomach toxicity of matrine on the lady beetle's life-table parameters, predatory ability, flight ability of parental adults, and cross-generational effects on life-table parameters of the predator's offspring. We found that matrine at 2,000 mg/l did not have any significant negative effects to adult fecundity, longevity, or the predatory capacity of parental adults of H. variegata. Moreover, it is the same for cross-generational effects of matrine on H. variegate. The contact toxicity of matrine significantly reduced the flight time of H. variegata males, but did not significantly affect flight time and average velocity. Our results support the view that matrine is safe to H. variegata and can be recommended for use in the local IPM strategy for control of A. gossipii.

The native parasitoids Bracon cephi (Gahan) and B. lissogaster Muesebeck (Hymenoptera: Braconidae) reduce populations of Cephus cinctus Norton (Hymenoptera: Cephidae), a native grassland species, and major wheat pest on the Northern Great Plains of North America. Non-host feeding adults of these braconids increase longevity, egg load, and egg volume when provisioned carbohydrate-rich diets. Nutrition from nectar can enhance the success of natural enemies in pest management programs. Cowpea, Vigna unguiculata (L.) Walpers, is a potential cover-crop that could add resilient features to the landscape and has extrafloral nectaries (EFN), easy-access nectar sources for beneficial insects. If more cowpea was grown on the Northern Great Plains, would B. cephi and B. lissogaster benefit from foraging on putatively beneficial EFN? We investigated cowpea inflorescence stalk extrafloral nectars (IS-EFN) and leaf stipel extrafloral nectars (LS-EFN) as potential food sources for these parasitoids. Females were caged on EFN sources on living cowpea plants to assess longevity. Egg load and volume were measured at 2, 5, and 10 days after placement. Bracon cephi survived 10 days on water, 38 days on IS-EFN; B. lissogaster 6 days on water, 28 days on IS-EFN. Bracon lissogaster maintained a constant egg load and volume across treatments while B. cephi produced 2.1-fold more eggs that were 1.6-fold larger on IS-EFN. Y-tube olfactometry indicated adult females were attracted to airstreams containing cowpea volatiles. These results demonstrate that non-native, warm-season cowpea benefits these native parasitoids and may improve conservation biocontrol of C. cinctus.

The redbanded stink bug, Piezodorus guildinii (Westwood, 1837), is a highly destructive soybean pest native to the Neotropical Region. In the past 60 yr, P. guildinii has been observed to expand its distribution in North and South America, causing significant soybean yield losses. In order to predict the future distribution direction of P. guildinii and create an effective pest control strategy, we projected the potential global distribution of P. guildinii using 2 different emission scenarios, Shared Socioeconomic Pathways 126 and 585, and 3 Earth system models, with the maximum entropy niche model (MaxEnt). Then, the predicted distribution areas of P. guildinii were jointly analyzed with the main soybean-producing areas to assess the impact for different soybean region. Our results showed that temperature is the main environmental factor limiting the distribution of P. guildinii. Under present climate conditions, all continents except Antarctica have suitable habitat for P. guildinii. These suitable habitats overlap with approximately 45.11% of the total global cultivated soybean areas. Moreover, P. guildinii was predicted to expand its range in the future, particularly into higher latitudes in the Northern hemisphere. Countries, in particular the United States, where soybean is widely available, would face a management challenge under global warming. In addition, China and India are also high-risk countries that may be invaded and should take strict quarantine measures.The maps of projected distribution produced in this study may prove useful in the future management of P. guildinii and the containment of its disruptive effects.

The governments of Australia and New Zealand require a phytosanitary treatment to control adult brown marmorated stink bug, Halyomorpha halys (Stål) (Hemiptera: Pentatomidae), as overwintering aggregations have been intercepted in the importation pathway of various nonhorticultural consignments, including vehicles. The commercialized fumigant, eFUME, a 16.7% by mass dilution of ethyl formate in carbon dioxide, resulted in complete control of ca. 1,000 field-collected, naturally diapausing adult H. halys in each of 3 independent trials at 10 ± 0.5 °C (x ± 2s) when ethyl formate levels in enclosure headspace were maintained steady-state at ca. 14.5 mg/liter for 4 h to yield Ct exposures ranging from 57.9 to 63.1 mg/liter h. Consistent with previous findings where greenhouse reared H. halys were controlled using laboratory formulations of this ethyl formate—carbon dioxide mixture, these confirmatory methods and results further inform technical and operational features of commercial practice.

The efficacy of an existing cold disinfestation postharvest treatment targeting Ceratitis capitata (Wiedemann) in citrus was determined for the marula fly, Ceratitis cosyra (Walker). The cold tolerances of C. capitata and C. cosyra were first quantified in artificial diet at 3.5 °C at different exposure periods for up to 18 days. Ceratitis capitata was found to be more cold tolerant than C. cosyra. At 3.5 °C, the duration to achieve 99.9968% mortality was calculated to be 11.57 days for C. capitata and 9.10 days for C. cosyra. Under an existing C. capitata cold treatment schedule at 1 °C for 14 days, the conditions required for complete mortality of the third larval stage of C. cosyra in orange, Citrus sinensis (L.) Osbeck cv. Valencia, were then determined. No survivors of C. cosyra in oranges were recorded beyond 11 days of cold treatment at 1 °C. The efficacy of this C. capitata treatment for disinfestation of C. cosyra was thereafter confirmed in large scale trials in Valencia oranges. In the large-scale trial at the lowest mean temperature of 1.19 °C for 14 days, there were no survivors from a total of 85 490 treated C. cosyra third instars in oranges. Since C. capitata was shown to be more cold tolerant than C. cosyra and a large scale test demonstrated at least 99.9965% efficacy after 14 days at 1.19 °C, compared with the established effective C. capitata cold treatment of 14 days at 1.11 °C, cold disinfestation treatments for C. capitata should be at least equally effective against C. cosyra.

The fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), is a significant invasive pest identified as a serious threat to global agricultural production and food security. However, its ovipositional preference, larval feeding selectivity, and nutritional indices are less studied. Here, we investigated these traits of FAW when fed on maize, wheat, soybean, tomato, cotton, and Chinese cabbage, and analyzed the correlation between its nutritional indices and the nutritional contents of crops. The results showed that the highest number of eggs were laid on maize and the lowest number were laid on tomato. The highest feeding choice rate of third instar larvae was on maize, and the lowest was on Chinese cabbage. The fifth instar larvae showed the highest feeding choice rate on maize, but no significant differences were found among other crop species. The food consumption (FC), the relative growth rate (RGR), and the approximate digestibility (AD) were significantly higher on maize and wheat, while the efficiency of conversion of ingested food (ECI) and the efficiency of conversion of digested food (ECD) were significantly higher when fed on cotton and Chinese cabbage. The FC, the relative consumption rate (RCR), RGR, and AD were significantly and positively correlated with soluble sugar and protein contents of host plants, while the ECI and ECD were significantly and negatively correlated with the soluble sugar content. The present study indicates that FAW may cause potential economic losses to these crops besides maize, and these findings are valuable in managing and controlling this pest.

Graphical Abstract

Chemical pesticides, while playing an important role in the suppression of insect pests, should be used in a manner that minimizes negative effects on natural enemies. The parasitoid, Anagyrus vladimiri Triapitsyn (Hymenoptera: Encyrtidae), plays an important role in the management of mealybug pests of citrus groves in the Mediterranean region. This study was conducted to evaluate the effect of commonly used acaricides (Spirodiclofen, Spirotetramat, Sulfur, Fenpyroximate, Abamectin) and mineral oils (Levanola, EOS, JMS, and Ultrapaz) on acute mortality of A. vladimiri. Toxicity was assessed in 4 cases: (i) direct spray application on adults, (ii) pesticide application on the mummified host, (iii) feeding with contaminated food, and (iv) contact with pesticide residue. The pesticide Abamectin, applied alone and with Levanola oil was highly toxic to adults in all bioassays, with the exception of direct spray application on the mummified host. Fenpyroximate was found to be highly toxic only when sprayed directly on adults, and sulfur was slightly harmful. Mineral oils were harmful when ingested with food; otherwise, they did not cause appreciable adult mortality. The findings of the present study suggest that all tested materials, with the exception of Abamectin and Fenpyroximate, are compatible with the survival of A. vladimiri. Direct ingestion of oils can, however, cause a degree of mortality. Given that indiscriminate use of these pesticides may affect the population ecology of A. vladimiri, they should be used with caution.

Graphical Abstract

Timing of insecticide application can impact efficacy, given variation in both weather and development of the crop and its insect pests. Both target and nontarget insects may vary in life stage and abundance at the time of application. In alfalfa Medicago sativa L. cropping systems, producers have interest in early-season insecticide applications to eliminate last-minute decisions about preharvest applications for alfalfa weevil Hypera postica (Gyllenhal) (Coleoptera: Curculionidae).The standard recommendation is based on scouting larvae close to the first harvest time. We compared early and standard timing of application of a lambda-cyhalothrin pyrethroid on alfalfa pest and beneficial insects. Field trials at a university research farm were conducted in 2020 and 2021. In 2020, early application was as effective as the standard timing against alfalfa weevil, as compared to the untreated control, but less effective than the standard timing in 2021. Effects of timing against Lygus bugs (Hemiptera: Miridae), grasshoppers (Orthoptera: Acrididae), and aphids (Hemiptera: Aphididae) were inconsistent between years. We observed the potential for early application to reduce negative impacts on ladybird beetles (Coleoptera: Coccinellidae) and spiders (Araneae), however, damsel bugs (Hemiptera: Nabidae) were similarly reduced by insecticide application regardless of timing. Overall arthropod community composition differed by both year and treatment. Future research should explore potential trade-offs of spray timing at larger spatial scales.

Nearly all maize seed sold in the United States includes a neonicotinoid seed treatment (NST), meant to protect seedlings against early-season insect pests. For key pests, including western corn rootworm (Diabrotica virgifera virgifera LeConte) (D.v.v), insecticidal proteins derived from Bacillus thuringiensis (Bt) are expressed in plant tissues as alternatives to soil-applied insecticides. Insect resistance management (IRM) plans use non-Bt “refuges” to encourage survival of Bt-susceptible D.v.v., which maintains susceptible alleles in the population. In non-cotton producing regions, IRM guidelines require a minimum 5% blended refuge for maize expressing more than 1 trait targeting D.v.v. Prior work has shown that 5% blends yield insufficient proportions of refuge beetles to contribute reliably to IRM. Whether NSTs interfere with survivorship of refuge beetles is unknown. Our objective was to determine whether NSTs affect proportions of refuge beetles, and secondarily, to determine whether NSTs provide agronomic advantages over Bt seed alone. To reveal host plant type (i.e., Bt or refuge), we used a stable isotope (¹⁵N) to mark refuge plants in plots with 5% seed blends. To assess refuge performance between treatments, we compared proportions of beetles from respective natal hosts. In all site-years, NSTs showed inconsistent effects on proportions of refuge beetles. Treatment comparisons showed inconsistent agronomic benefits of NSTs when combined with Bt traits. Our results demonstrate that NSTs have a negligible impact on refuge performance and reinforces the assertion that 5% blends are serving little benefit for IRM. Plant stand and yield were not improved by NSTs.

The Nantucket pine tip moth (NPTM) (Rhyacionia frustrana Comstock) is a native, regeneration pest of young loblolly pines (Pinus taeda L.), causing shoot dieback, tree deformity, and growth and volume declines. Soil applications of systemic insecticides may be an effective strategy to suppress NPTM populations. The study objective was to assess the efficacy of four systemic insecticide treatments (chlorantraniliprole, dinotefuran, fipronil, and imidacloprid) for two growing seasons in outplanted bareroot and containerized seedling trials. Response variables included NPTM infestation rates, along with tree height, groundline diameter, volume index, and stem form. Infestation rates significantly decreased for each systemic insecticide treatment during the first year compared to controls, although dinotefuran and imidacloprid provided season-long control in one trial. Chlorantraniliprole reduced NPTM infestation rates for two growing seasons in both trials. While imidacloprid treatments did not alter growth metrics except for one comparison, fipronil and dinotefuran treatments improved several growth metrics. Chlorantraniliprole consistently improved growth metrics throughout the study.

The emerald ash borer (EAB), Agrilus planipennis Fairmaire, is the most destructive invasive pest on ash (Fraxinus spp.) in the United States. We determined whether ash trees injected with emamectin benzoate (EB) could protect untreated neighboring ash trees. We also determined whether the selective treatment of ash trees with EB injections had adverse effects on the establishment of introduced larval parasitoids Tetrastichus planipennisYang and Spathius galinae Belokobylskij & Strazenac. In experiment one, trees were treated with EB and then retreated 3 years later. Five years post initial treatment, we found that 90% of treated ash trees retained healthy crowns, significantly higher than those of untreated control ash trees (16%). For experiment two, trees only received one treatment of EB and after 2 years 100% of treated ash trees retained healthy crowns, significantly higher than those of untreated ash trees (50%). In both experiments, we found that distance from the central EB-treated tree was not a significant predictor for tree health or presence of EAB exit holes. Although distance from the EB-treated trees appeared to have a significant positive relationship with woodpecker feeding signs on neighboring trees, such relationships did not result in significant differences in the proportion of neighboring ash trees retaining healthy crowns between EB treatment and control plots. The introduced EAB parasitoids appeared to have established equally well between treatment and control plots. Findings are discussed in the context of integration of EB trunk injection with biological control for protection of North American ash against EAB.

The use of toxic baits has become one of the main methods of management of fruit flies in Brazil.The application of toxic baits may cause side effects on the native parasitoid Doryctobracon areolatus (Hymenoptera: Braconidae). Based on the results, formulations made from the food attractants 3% Biofruit, 1.5% Ceratrap, 1.25% Flyral, 3% Isca Samaritá, 3% Isca Samaritá Tradicional, and 7% sugarcane molasses associated with the Malathion 1000 EC and the ready-to-use toxic bait Gelsura (containing the active ingredient alpha-cypermethrin) were classified as harmful (class 4) to D. areolatus (mortality > 85% at 96 HAE). In contrast, for toxic baits formulated with insecticide phosmet, the mortality ranged from 38% to 72%, classified as slightly harmful or moderately harmful. However, when phosmet was added to the 3% SamaritáTradicional bait, the mortality was only 3.9% (class 1—harmless), similar to the toxicity observed for the Success 0.02 CB ready-to-use bait (0.24 g a.i. spinosad/l) (<5% mortality). Although toxic baits were formulated with spinosyn-based insecticides, all toxic bait formulations were classified as harmless or slightly harmful (<50% mortality) to D. areolatus, with the exception of 1.5% Ceratrap + spinetoram and 7% Sugarcane molasses + spinosad (≈ 60% mortality—moderately harmful). In addition, these formulations did not show sublethal effects in reducing the parasitism and emergence rate of the F₁ generation of D. areolatus in A. fraterculus larvae. The results serve as a basis for the correct use of toxic food baits without affecting the biological control.

Rugose spiraling whitefly (RSW), Aleurodicus rugioperculatus Martin, a native pest of coconut in Central America, has recently been introduced to South-East Asia. Little is known about the spread of RSW in Bangladesh, the effect this pest has on coconut plants, or the full range of its plant hosts. To fill this knowledge gap, we conducted surveys across the southern coastal coconut producing region of Bangladesh. Coconut plant damage from RSW was high throughout all the areas surveyed, and this was correlated with the population density of RSW. Areas, such as Khulna, which had the highest population density of RSW, had the highest level of RSW leaf damage. Coconut plants with an infestation of RSW had significantly fewer leaves and fruits. The coconut fruits were also smaller, both in terms of size and weight, and contained less water and lighter endosperm compared to non-infested plants. This highlights the need for the management of RSW in coconut growing regions. We also report 39 new host plant species for RSW from 22 plant families, including the economically important mangrove plant Nipa palm, Nypa fruticans Wurmb.

Due to declines in pollinator populations, many people are now interested in learning about which annual flowers they can plant in their garden to better support pollinators. However, reports of experimental evaluation of cultivars of annual flowers for attraction to pollinators are scarce. We sampled pollinators visiting six cultivars of marigold (Tagetes erecta and T. patula), ten cultivars of bidens (Bidens ferulifolia and B. aurea), and eight cultivars of portulaca (Portulaca oleracea and P. grandiflora) for two years to compare pollinator visitation rates among cultivars within each flower type. Pollinators collected on flowers in research plots were categorized into four groups, honey bees (Apis mellifera), common eastern bumble bees (Bombus impatiens), wild bees, and syrphids, to show the proportion of different pollinator visitors to each cultivar. Pollinator visitation rates varied significantly among cultivars of marigold, bidens, and portulaca, with some cultivars having as much as 10-fold the visitation rate of other cultivars of the same flower type. In the second year we also evaluated nectar production and nectar quality of the most and least visited cultivars of portulaca and bidens. Our results show that pollinators have a strong preference for cultivars that produce the most nectar or nectar with the highest sugar content. This research will better inform entomologists, growers, educators, and plant breeders, about which cultivars of marigold, portulaca, and bidens are visited the most by pollinators, and how to accurately determine this at the cultivar level.

Amyelois transitella is the primary pest of pistachios in California. The first A. transitella outbreak of the twenty-first century occurred in 2007 and a total of five outbreaks occurred between 2007 and 2017 (total insect damage >1%). This study used processor information to identify the nut factors associated with the outbreaks. Processor grade sheets were used to explore the relationship between the variables time of harvest, percent nut split, percent nut dark staining, percent nut shell damage, and percent adhering hull for Low Damage (82,537 loads) and High Damage years, (92,307 loads). Total insect damage (±SD) for the Low Damage years averaged 0.005 ± 0.01 and in High Damage years was three times higher, 0.015 ± 0.02. In Low Damage years the strongest correlation was between total insect damage and two variables, percent adhering hull and dark stain (0.25, 0.23) while in High Damage years the correlation between total insect damage and percent dark stain was the highest (0.32) followed by percent adhering hull (0.19). The linkage of these nut factors to insect damage suggests that outbreak prevention depends on early identification of premature hull split/breakdown in addition to the traditional emphasis on treating the standing population of A. transitella.

Nipaecoccus viridis (Newstead) (Hemiptera: Pseudococcidae) is an emerging invasive pest of citrus. There is no information on the life history of N. viridis in relation to citrus cultivars produced in Florida. Here, we quantified the life-history parameters of N. viridis on 6 citrus and 3 non-citrus host plant species under laboratory conditions. We also investigated the effect of mode of reproduction (sexual vs. parthenogenic) on the fecundity of N. viridis. Nipaecoccus viridis was able to develop to maturity and reproduce on all host plant taxa tested. The individual host plants affected the developmental and survival rate, adult longevity, and offspring production of N. viridis. All citrus genotypes commercially grown in Florida were susceptible to N. viridis, with the exception of Citrus aurantium L., which suggests that there is need for management of this invasive pest in Florida citrus groves. Murraya paniculata L., Hibiscus rosa-sinensis L., and Punica granatum L. were also acceptable hosts for N. viridis. Sexually reproducing female N. viridis produced significantly more eggs than those reproducing asexually. Based on our results, propagation of M. paniculata and other citrus relatives should be discouraged around citrus groves in Florida. Given the fitness benefit of sexual reproduction in this pest, pheromone-based tools like mating disruption and mass trapping of adult males could be considered as part of an integrated pest management program with insecticides.

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.

The traditional stake survey and in-ground (IG) monitoring stations have been ineffective in aggregating the Asian subterranean termite, Coptotermes gestroi (Wasmann) in southeastern Florida. In this study, we used both IG and above-ground (AG) Sentricon stations to monitor and bait C. gestroi, and as expected, none of the 83 IG stations was intercepted. Despite this, AG bait stations with 0.5% noviflumuron were successfully used to eliminate C. gestroi colonies. From 2 field experiments, the mean colony elimination time (±SD) using AG baits were 6.4 ± 3.8 wk (n = 4) and 8.0 ± 2.1 wk (n = 12), respectively. Such results were compatible with baiting studies against field colonies of C. gestroi elsewhere, that is, 4–9 wk. The successful rates in monitoring and baiting of C. gestroi with IG stations in other regions also varied, which may be due to the variabilities in tunnel geometry of this species in different environments. In areas with established C. gestroi populations, routine inspection for signs of activity in structures and surrounding trees can be a critical component for pest control providers for early detection of infestation and colony elimination with AG bait stations.

The first case of field-evolved resistance in European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) to transgenic corn (Zea mays L.) producing a Bacillus thuringiensis (Bt) Berliner toxin was discovered in Nova Scotia, Canada in 2018. This case involved resistance to Bt corn producing Cry1Fa toxin. As a mitigation response, Bt corn hybrids producing only Cry1Fa were replaced in that region with hybrids producing two or three Bt toxins targeting O. nubilalis. In this study, we collected O. nubilalis in several corn-growing regions of Canada during 2018 to 2020 and tested their progeny for susceptibility to four Bt toxins produced by currently available Bt corn that targets O. nubilalis: Cry1Fa, Cry1Ab, Cry1A.105, and Cry2Ab. Based on toxin concentrations killing 50% of larvae from 23 field-derived strains relative to two susceptible laboratory strains, the resistance ratio was at least 10 for Cry1Fa for 12 strains (52% of strains) consisting of 10 strains from Nova Scotia, as well as strains from near Montreal, Quebec and Roseisle, Manitoba. We found low but statistically significant resistance relative to at least one of two susceptible strains for Cry1Ab (23% of strains), Cry1A.105 (45% of strains), and Cry2Ab (14% of strains), with maximum resistance ratios of 3.9, 5.8, and 2.0, respectively. These results provide key information for addressing O. nubilalis resistance to Bt corn in Canada.

The objective of this study was to evaluate the physicochemical compatibility of mixtures of synthetic and botanical limonoid-based insecticides, as well as the toxicity of these associations, in the management of Spodoptera frugiperda (J.E. Smith) under laboratory and field conditions. For this, the associations of 4 commercial botanical insecticides based on neem registered in Brazil (Azamax, Agroneem, Azact CE, and Fitoneem) were tested with synthetic insecticides from the group of growth regulators (IGRs [triflumuron, lufenuron, methoxyfenozide and tebufenozide]). When mixed, all combinations caused a significant reduction in the pH of the mixture and a significant increase in electrical conductivity. However, all tested combinations showed similar stability behavior to the negative control (distilled water), which demonstrated their physicochemical compatibility. Furthermore, in laboratory and field bioassays, mixtures of IRGs with limonoid-based formulations provided satisfactory effects in the management of S. frugiperda. However, binary mixtures of insecticide Intrepid 240 SC with Azamax or Azact CE (at LC₂₅ previously estimated) showed the highest toxicities on S. frugiperda larvae in laboratory bioassays and damage reduction caused by S. frugiperda in a 2-yr field experiments. Therefore, mixtures of IGRs with limonoid-based botanical insecticides are promising alternatives for the management of S. frugiperda and important component of integrated pest management and insect resistance management programs.

The fall armyworm, Spodoptera frugiperda (J. E. Smith), is a highly polyphagous pest native to the tropical Americas that has recently spread to become a global super-pest threatening food and fiber production. Transgenic crops producing insecticidal Cry and Vip3Aa proteins from Bacillus thuringiensis (Bt) are used for control of this pest in its native range. The evolution of practical resistance represents the greatest threat to sustainability of this technology and its potential efficacy in the S. frugiperda invasive range. Monitoring for resistance is vital to management approaches delaying S. frugiperda resistance to Bt crops. DNA-based resistance screening provides higher sensitivity and cost-effectiveness than currently used bioassay-based monitoring. So far, practical S. frugiperda resistance to Bt corn-producing Cry1F has been genetically linked to mutations in the SfABCC2 gene, providing a model to develop and test monitoring tools. In this study, we performed targeted SfABCC2 sequencing followed by Sanger sequencing to confirm the detection of known and candidate resistance alleles to Cry1F corn in field-collected S. frugiperda from continental USA, Puerto Rico, Africa (Ghana, Togo, and South Africa), and Southeast Asia (Myanmar). Results confirm that the distribution of a previously characterized resistance allele (SfABCC2mut) is limited to Puerto Rico and identify 2 new candidate SfABCC2 alleles for resistance to Cry1F, one of them potentially spreading along the S. frugiperda migratory route in North America. No candidate resistance alleles were found in samples from the invasive S. frugiperda range. These results provide support for the potential use of targeted sequencing in Bt resistance monitoring programs.

Foliar-applied insecticides are commonly used for adult western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), control in Nebraska but little efficacy data is available. Anecdotal reports of reduced efficacy in areas of northeast Nebraska led to the conduct of this study. Objectives were to (i) evaluate the efficacy of commercial applications of commonly used formulated insecticides (bifenthrin, lambda-cyhalothrin, chlorpyrifos, or tank mixes) for WCR control in 7 northeast Nebraska counties during 2019 and 2020 and (ii) conduct adult WCR concentration-response vial bioassays with bifenthrin, chlorpyrifos, and dimethoate active ingredients on a subset of field populations. Whole plant counts (WPC) were used to measure WCR densities in insecticide-treated and untreated maize fields before and after insecticide application. Field control was excellent with organophosphate/pyrethroid tank mixes as proportional change in mean WPC of treated fields was significantly reduced (>0.90) versus untreated fields where little change in WPC occurred. The exception was one treated Boone County field where proportional reduction in WPC was ≤0.78. Bioassays revealed LC₅₀s and resistance ratios of most populations exposed to bifenthrin and dimethoate were not significantly different than the susceptible control. Most populations exhibited a low level of chlorpyrifos resistance when compared to the susceptible control. Field and lab data suggest the local onset of practical WCR field-evolved resistance to bifenthrin in Boone County and chlorpyrifos in Boone and Colfax counties. Results of this study will increase our understanding of WCR resistance evolution, serve as a comprehensive baseline for future research, and inform WCR management programs.

Bactrocera tsuneonis and Bactrocera minax are the most destructive pests that damage citrus in China. These key pests hinder the citrus trade, cause significant financial losses, drastically lower citrus production and quality, and decrease farmer enthusiasm for citrus planting. Bactrocera minax and B. tsuneonis are very similar in all life stages. There are limited morphological characteristics to differentiate the adult species, and it is nearly impossible to differentiate these two species in the egg and larval stages. Loop-mediated isothermal amplification (LAMP) is a rapid and robust diagnostic tool used to identify these two species accurately. We designed two sets of primers to distinguish B. minax and B. tsuneonis using DNA barcoding region of the COI gene. Only 50 min was needed under a constant temperature of 65°C to determine the species of the two flies. The reaction system has high specificity and sensitivity, in which these two species can be accurately distinguished between different geographical populations and 1.0 ng/µL was the lowest DNA concentration that could be detected. Our primers can quickly identify these key pests without knowing their morphology, which could facilitate plant protection workers at the primary level to solve problems in plant quarantine.

As the most outer layer between itself and the environment, integuments are necessary for insects with various important functions. Cuticular proteins (CPs) are the main components in integuments, while the functions of CP genes remain unknown in Mythimna separata (Walker), which is a devastating agricultural pest. In this study, 79 CP genes were identified from the transcriptomes of larval integuments, 57 of which were from the family containing conserved Rebers & Riddiford (R&R) consensus (CPR family). Amongst these CPRs, 44 genes belonged to the subfamily with RR-1 motif (RR-1 genes) and clustered into three clades, with the top 15 most abundant RR-1 genes identified based on fragments per kilobase per million mapped fragments (FPKM) values. RT-qPCR analysis showed that most of RR-1 genes such as MsCPR1-4 were highly expressed at larval stages and in their integuments. The expression levels of RR-1 genes were generally decreased at the beginning but increased at the late stage of molting process. RNAi was applied for six RR-1 genes, and MsCPR1-4 were knocked down significantly. Silence of MsCPR2 resulted in abnormal integument formed after molting, while knockdown of MsCPR3 and MsCPR4 led to failure of molting, respectively. No phenotype was obtained for the RNAi of MsCPR1. Therefore, the expression of RR-1 genes and their functions were analyzed in the development of integuments in M. separata, providing new insights of RR-1 genes and potential targets for the development of growth regulators and new insecticides for M. separata.

Graphical Abstract

Helicoverpa armigera (Hübner) is a major crop pest native to Europe, Asia, Australia, and Africa which has recently invaded South America and has caused billions of dollars in agricultural losses. Because of challenges in differentiating between H. armigera and Helicoverpa zea (Boddie), a closely related species native to North and South America, genetic tests have previously been developed to detect H. armigera DNA in pooled samples of moth legs. In this study, a field-based recombinase polymerase amplification (RPA) assay using a lateral flow strip and a qPCR melt curve assay were developed for specific detection of H. armigera DNA in pooled moth samples. In addition, a crude DNA extraction protocol for whole moths was developed to allow rapid preparation of DNA samples.The RPA field test was able to detect ≥ 10 pg of purified H. armigera DNA and the crude DNA of one H. armigera sample in a background of 999 H. zea equivalents. The qPCR assay was able to detect ≥ 100 fg of purified H. armigera DNA and the crude DNA of one H. armigera sample in a background of up to 99,999 H. zea equivalents. Both RPA and qPCR assays detected H. armigera in the crude DNA extracted in the field from a pool of one H. armigera moth and 999 H. zea moths. These newly developed molecular assays to detect H. armigera will contribute to large-scale surveillance programs of H. armigera.

The fall armyworm (FAW) Spodoptera frugiperda was first found in China in 2018. In other countries, FAW has evolved corn and rice strain biotypes. It is not possible to identify these strains based on morphology. In addition, FAW is very similar in appearance to several other common pests. These situations bring great challenges to the population management of FAW. In this study, we developed a rapid identification method based on PCR-RFLP to distinguish the two FAW strains and the FAW from other lepidopteran pests. A 697 bp mitochondrial cytochrome c oxidase I (COI) was cloned and sequenced from FAW, Spodoptera litura, Spodoptera exigua, and Mythimna separata. The COI fragments of these species revealed unique digestion patterns created by three enzymes (Tail, AlWN I, and BstY II). Thus, these four species can be distinguished from each other.The enzyme Ban I recognized a unique SNP site on a 638 bp triosephosphate isomerase (Tpi) fragment of the corn strain FAW. The Tpi fragment of the corn strain was cut into two bands. However, the rice strain could not be digested. Using this method, all 28 FAW samples collected from different host plants and locations in China were identified as the corn strain.This suggests that the rice strain has not yet invaded China. This method allows discrimination of FAW from other Lepidopteran pests and distinguishes the two FAW host strains.

Graphical Abstract

The tomato pest Phthorimaea absoluta Meyrick is highly invasive but has not yet invaded North America. However, several morphologically similar species are already present, making detection of P. absoluta presence and invasion challenging. We designed a quantitative PCR molecular diagnostic to differentiate P. absoluta, P. operculella (Zeller), or Keiferia lycopersicella (Walsingham) (Lepidoptera: Gelechiidae) DNA. Additionally, we developed an RPA-Cas12a molecular diagnostic that allows for the isothermal detection of P. absoluta DNA, eliminating the need for a thermocycler. The results of the RPA-Cas12a diagnostic can be visualized simply using a UV light source and cell phone camera. We expect these diagnostics to improve quarantine and prevention measures against this serious agricultural threat.

Typical delimitation trapping survey designs for area-wide (nonlocalized) insect populations are regularly spaced grids, and alternative shapes have not been evaluated. We hypothesized that transect-based designs could give similar detection rates with significantly shorter servicing distances. We used the TrapGrid model to investigate novel “trap-sect” designs incorporating crossed, spoked, and parallel lines of traps, comparing them to a regular grid, in single survey and multiple-site scenarios. We calculated minimum servicing distances and simulated mean probabilities of detecting a pest population, judging overall performance of trap network designs using both metrics. For single sites, trap-sect designs reduced service distances by 65–89%, and most had similar detection probabilities as the regular grid. Kernel-smoothed intensity plots indicated that the best performing trap-sect designs distributed traps more fully across the area. With multiple sites (3 side by side), results depended on insect dispersal ability. All designs performed similarly in terms of detection for highly mobile insects, suggesting that designs minimizing service distances would be best for such pests. For less mobile pests the best trap-sect designs had 4–6 parallel lines, or 8 spokes, which reduced servicing distances by 33–50%. Comparisons of hypothetical trap-sect arrays to real program trap locations for 2 pests demonstrated that the novel designs reduced both trap numbers and service distances, with little differences in mean nearest trap distance to random pest locations. Trap-sect designs in delimitation surveys could reduce costs and increase program flexibility without harming the ability to detect populations.

Two insecticides (deltamethrin and pirimiphos-methyl) were evaluated in laboratory assays as grain protectants when applied as a total percentage of a maize mass to control adult Prostephanus truncatus and Sitophilus zeamais. All experiments were conducted at the University of Thessaly in Greece at 30°C and 65% RH under continuous darkness. Either insecticide was applied to 20 g of maize placed in a vial or to the upper one half, one fourth, or one-eighth layer of the maize, then insects were either added to the vials before or after the maize in a completely randomized block design CRBD with n = 9 replicates. Mortality, progeny production, and insect damaged kernels (IDK) were then evaluated for each vial. Insect introduction method (before or after) did not have any impact on any of the variables. Mortality was nearly 100% for all treatments for both insecticides for P. truncatus. Subsequently, progeny production and the number of insect damaged kernels were very low or zero for P. truncatus. Mortality of S. zeamais remained low across layer treatments for deltamethrin. However, S. zeamais was easily controlled by pirimiphos-methyl. The results of this laboratory study show that while deltamethrin and pirimiphos-methyl have some effectiveness as a layer treatment on a column of maize, efficacy will be dependent on the target species, and the depth of the treated layer, as well as the location on which the insects are present.

Graphical Abstract

Onion thrips (Thrips tabaci Lindeman,Thysanoptera:Thripidae) causes severe damage to many horticultural and agronomic crops worldwide. It also acts as a vector of several plant viruses. T. tabaci is a key pest of Allium cepa in the United States. However, there is limited information available on the genetic variation within and between T. tabaci populations in the United States and its key evolutionary parameters. In the current study, 83 T. tabaci specimens were collected from A. cepa from 15 different locations comprising four states of the United States. A total of 92 mtCOI gene sequences of T. tabaci from A. cepa were analyzed to understand the genetic diversity and structure of T. tabaci collected from onion host. Seven distinct haplotypes of T. tabaci infesting A. cepa were identified from the current collection, while nine T. tabaci sequences retrieved from GenBank comprised 5 haplotypes. Overall, 15 haplotypes of T. tabaci infesting A. cepa were identified in the world that includes the ten haplotypes in the United States. In the phylogenetic analysis, all the populations collected during the study clustered with thelytokous lineage, while T. tabaci sequences retrieved from GenBank corresponded to leek-associated arrhenotokous lineage. The highest genetic variation was found in Elba and Malheur populations with 3 haplotypes identified in each. The results suggest that haplotypes 1 and 7 are more frequently prevailing haplotypes in the north-western United States, with haplotype 1 being the predominant all over the country. The eastern United States appears to have a more diverse group of haplotypes. The populations from Hungary constituted distinct haplotypes and a haplotype from Kingston linked it with the predominant haplotype.

Reticulitermes flavipes is the most invasive species in its genus and is responsible for causing significant damage to human structures in areas where it has been introduced. Although it has already become established in Chile and Uruguay, it had not previously been reported in Argentina. In this study, we report the first detection of this species in Buenos Aires, the capital of Argentina. The colony was already producing alates, and species identification was confirmed through both morphology and mitochondrial gene 16S rRNA analysis. Our results, while not conclusive, suggest that this introduction was independent from the one that occurred in Chile and Uruguay, and potentially originated from the United States. The detection of R. flavipes in Argentina is significant because it highlights the potential for this species to establish itself in new regions and underscores the need for future research on and control of R. flavipes in this country.