Effects of nonnutritive sugar inclusion in laboratory diets and attracticidal spheres on survivorship and mobility of 2 Dipteran species, Rhagoletis pomonella (Diptera: Tephritidae) and Drosophila suzukii (Diptera: Drosophilidae).

Native apple maggot fly, Rhagoletis pomonella, and invasive spotted-wing drosophila, Drosophila suzukii, are key pests of apple and small fruit, respectively, in the United States. Both species are typically managed with standard insecticide applications. However, interest in alternative strategies that result in insecticide reductions has led to evaluations of nonnutritive sugars as toxicants for Drosophila species and development of attracticidal spheres for both species. Here, we evaluated the survivorship of R. pomonella and D. suzukii when provided with standard diets that substituted saccharin, sucralose, aspartame, erythritol, dextrose, or mannitol for the sucrose component and compared them with standard diets and water-only controls for up to 15 days. Presence of erythritol and mannitol significantly decreased survivorship of R. pomonella and erythritol significantly decreased the survivorship of D. suzukii. However, mobility trials following a 2 h exposure to aqueous solutions of each sugar treatment resulted in no strong impact on either species. Survivorship after 30 min exposure to erythritol or mannitol alone, or in combination with varying concentrations of sucrose (serving as a phagostimulant) at 30 min and 24 h were evaluated for both species. Only D. suzukii survivorship was affected with decreased survivorship on erythritol:sucrose solutions of 20:0% and 15:5% for 24 h. Based on all results, erythritol appeared most promising, and was integrated into attracticidal spheres as a toxicant but even at the highest concentration, survivorship remained unaffected for either species, thus making this nonnutritive sugar impractical and ineffective as a toxicant substitute in attracticidal spheres.

Landscape-scale spatiotemporal dynamics of Halyomorpha halys (Stål) (Hemiptera: Pentatomidae) populations: implications for spatially-based pest management.

BACKGROUND: Brown marmorated stink bug (BMSB), Halyomorpha halys (Stål), is an invasive and severe pest of specialty and row crops. A 2-year field study conducted in four Mid-Atlantic states in the USA characterized the spatial and temporal dynamics of BMSB populations and its association with landscape elements in commercial agriculture settings. In each state, two 1 km2 sites included typical landscape elements (i.e., tree fruit orchards, annual field and vegetable crops, woodlands, and human-made structures). Twenty-seven georeferenced pheromone traps were deployed per site and the number of BMSB adults and nymphs captured was counted throughout the growing season. RESULTS: Findings from spatial analysis by distance indices, along with time-series maps of BMSB distribution, showed that BMSB exhibited significant spatial aggregation, and that its distribution was spatially consistent between years. Analyses with geographic information systems (GIS) revealed that BMSB 'hot spots' occurred in different landscape elements throughout each season. Most patches (i.e., clusters of significantly higher trap captures) were found near woodlands early in the season, near tree fruit orchards in summer, and on the border of annual field crops in autumn. Buffer analysis with GIS indicated that more BMSB adults were captured closer to woodlands compared with other landscape elements. CONCLUSION: Understanding the spatial and temporal movement and distribution of BMSB is critical to predicting their potential impact and ultimately devising strategies to mitigate this risk to vulnerable crops. The results of this study can be used to design streamlined, spatially-based areawide management of BMSB in heterogeneous and complex agricultural landscapes. © 2023 Society of Chemical Industry. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Development and survivorship of Lycorma delicatula (Hemiptera: Fulgoridae) on cultivated and native Vitis spp. (Vitales: Vitaceae) of the Eastern United States.

As Lycorma delicatula (White) continues to spread across the United States, more winegrapes are potentially susceptible to damage from this pest. Lycorma delicatula, spotted lanternfly, is primarily associated with Ailanthus altissima (Mill.) Swingle, a tree from its native range that is now globally distributed. While L. delicatula is a known pest of cultivated Vitis spp. in South Korea, its relationship with the specific grape species grown in the United States is unclear. This study assessed L. delicatula survivorship and development on 5 Vitis species, including 2 winegrape V. vinifera L. varieties, 'Pinot Noir' and 'Chardonnay', Concord grape, Vitis labrusca L., River grape, Vitis riparia Michx., and muscadine grape, Vitis rotundifolia Michx. var. 'Carlos'. A diet of A. altissima served as a positive control. Lycorma delicatula provided with a diet of V. riparia or V. vinifera 'Pinot Noir' yielded the highest survivorship and fastest rates of development among grape diets and were statistically equivalent to those provided with A. altissima. Vitis rotundifolia did not support L. delicatula growth past the third-instar life stage, indicating this species is a poor host for the early development of this pest. Our results indicate that both V. riparia and V. vinifera are favorable hosts for L. delicatula and may provide the means for this insect to invade and establish in new regions.

Do adult Magicicada (Hemiptera: Cicadidae) feed? Historical perspectives and evidence from molecular gut content analysis.

The periodical cicadas in the genus Magicicada are remarkable for their unusual life histories and dramatic synchronized emergences every 13 or 17 years. While aspects of their evolution, mating behaviors, and general biology have been well-characterized, there is surprising uncertainty surrounding the feeding habits of the short-lived adult stage. Despite a tentative scientific consensus to the contrary, the perception that adult Magicicada do not feed has persisted among the general public, and recent studies are lacking. We directly investigated the feeding behavior of Magicicada spp. through high-throughput sequencing (HTS)-based dietary analysis of nymphs, freshly molted (teneral) adults, and fully sclerotized adults collected from orchard and wooded habitats during the 2021 emergence of Brood X. Identifiable plant DNA (trnF, ITS amplicons) was successfully recovered from nymphs and adults. No plant DNA was recovered from teneral adults, suggesting that all DNA recovered from sclerotized adults was ingested during the post-teneral adult stage. Both nymphs and adults were found to have ingested a range of woody and herbaceous plants across 17 genera and 14 families. Significantly more plant genera per individual were recovered from adults than from nymphs, likely reflecting the greater mobility of the adult stage. We hypothesize that the demonstrated ingestion of plant sap by Magicicada adults is driven by a need to replace lost water and support specialized bacteriome-dwelling endosymbionts that cicadas depend upon for growth and development, which constitutes true feeding behavior.

Short- and long-term effects of season-long infestation of Lycorma delicatula (Hemiptera: Fulgoridae) on young apple (Malus domestica) and peach (Prunus persica) trees.

Lycorma delicatula White, commonly called spotted lanternfly, is an invasive fulgorid that was first detected in the United States in Pennsylvania in 2014 and has spread across Eastern states. Lycorma delicatula is a phloem feeder, and feeding of persistent populations can cause wilt and dieback in host plants. Vineyards in infested regions have experienced vine loss, where damage has been attributed to L. delicatula. Nymphal and adult lifestages have been observed on cultivated apple and peach trees, but the feeding effects of this pest on these crops have not been quantified. Here, we caged young potted apple and peach trees and infested them with 50 nymphal or 25 adult L. delicatula per plant for 3 months and tracked plant health. Horticultural measurements including trunk diameter, height, leaf drop, and photosynthesis rates were recorded before, during, and after the infestation. To evaluate the long-term effects of infestation, all apple and peach trees were removed from pots and planted in an orchard block where their phenology, growth, and physiology could be evaluated the following growing season. Short- and long-term measurements showed no significant differences in apple tree health between infested and noninfested trees. There was a significant reduction in the growth of trunk diameter in peach trees during exposure to early-instar nymphs; infested peach trees also had significantly increased frost damage to buds the following spring. However, there were no long-term impacts on peach growth and physiology. These results indicate that L. delicatula likely will not be a major threat to these deciduous fruit crops.

Halyomorpha halys (Hemiptera: Pentatomidae) trap captures at orchard and nonorchard sites and the influence of uncultivated woody host plants in adjoining woodlots.

The invasive Halyomorpha halys (Hemiptera: Pentatomidae) has threatened Mid-Atlantic tree fruit since 2010. To identify factors underlying observed differences in H. halys pest pressure among individual orchards within a geographically proximate area, a 3-yr study was conducted across 10 apple orchard and 8 nonorchard sites bordered by unmanaged woodlots. At each site, 3 pheromone traps were monitored weekly for H. halys captures from late April to mid-October. Apple injury was assessed at harvest at orchard sites annually, and a survey of woody plants found in woodlots adjacent to all sites was conducted. There were no significant differences in captures between orchard and nonorchard site types, but captures were significantly different among individual orchard sites and among individual nonorchard sites. A significant positive relationship between the amount of stink bug injury on apple at harvest and late season captures was detected at orchard sites in 2018 and 2019. Among woodlots adjacent to all sites, a significant positive relationship between the proportion of Lonicera spp. and mid- and late-season nymphal captures was identified. Season-long nymphal captures were positively related to the proportion of Lonicera and Elaeagnus and negatively with Sassafras. For adults, captures were negatively related to the proportion of Ailanthus and positively related to the proportion of Fraxinus in the early and mid-season, respectively. Our results indicate that orchard presence was not driving the relative abundance of localized H. halys populations and that differences in relative densities among sites point to other factors, such as abundance of specific uncultivated woody hosts in unmanaged areas.

Assessing acceptability of wild and cultivated hosts of Lycorma delicatula (Hemiptera: Fulgoridae) under semifield conditions.

Lycorma delicatula White (Hemiptera: Fulgoridae) is an invasive phloem feeder with a broad host range that includes both hardwood trees and cultivated temperate fruit crops. Here, we evaluated acceptability of wild hosts, A. altissima and Juglans nigra L. (Fagales: Juglandaceae) and cultivated hosts Vitis vinifera, Malus domestica (Rosales: Rosaceae), and Prunus persica L. Batsch (Rosales: Rosaceae) to L. delicatula under field conditions. Fluorescent-marked early instar nymphs, late instar nymphs, or adult L. delicatula were released at the base of single potted host plants and the number of individuals retained was recorded over 24 h. Paired choice trials with A. altissima and another host plant were conducted. Individuals retained on or moving between plants were recorded over 24 h. Sentinel A. altissima, J. nigra, V. vinifera, M. domestica, and P. persica potted plants were deployed at 5 sites and the number of L. delicatula present on each plant was recorded weekly. In single and paired host trials, early instars and adults were generally retained in higher numbers on A. altissima and V. vinifera, and late instars were retained on A. altissima and J. nigra. Significantly more L. delicatula were present on sentinel A. altissima compared with other host plants, except J. nigra during the period when late instars were the most prevalent lifestage in the field. These results indicate that wild hosts such as A. altissima and J. nigra are likely supporting establishment of L. delicatula populations, and that presence of cultivated V. vinifera may contribute to population establishment and growth.

Methyl salicylate improves the effectiveness of the odor-baited trap tree approach for adult plum curculio, Conotrachelus nenuphar (Coleoptera: Curculionidae), monitoring and attract-and-kill.

In commercial apple orchards, the odor-baited trap tree approach involving the synergistic lure composed of benzaldehyde (BEN) and the PC aggregation pheromone grandisoic acid (GA) serves as an effective monitoring tool as well as an attract-and-kill strategy for plum curculio (PC), Conotrachelus nenuphar Herbst. (Coleoptera: Curculionidae), management. However, the relatively high cost of the lure and the degradation of commercial BEN lures by UV light and heat discourage its adoption by growers. Over a 3-yr period, we compared the attractiveness of methyl salicylate (MeSA), either alone or in combination with GA, to plum curculio (PC) with that of the standard combination of BEN + GA. Our main goal was to identify a potential replacement for BEN. Treatment performance was quantified using 2 approaches: (i) unbaited black pyramid traps (2020, 2021) to capture PC adults and (ii) PC oviposition injury (2021, 2022) on apple fruitlets of trap trees and of neighboring trees to assess potential spillover effects. Traps baited with MeSA captured significantly more PCs than unbaited traps. Trap trees baited with a single MeSA lure and 1 GA dispenser attracted a similar number of PCs as trap trees baited with the standard lure composed of 4 BEN lures and 1 GA dispenser based on PC injury. Trap trees baited with MeSA + GA received significantly more PC fruit injury than neighboring trees suggesting no or limited spill-over effects. Our collective findings suggest that MeSA is a replacement for BEN thereby cutting costs of lures by ca. 50% while maintaining trap tree effectiveness.

Confirmatory evaluation of eFUME for control of brown marmorated stink bug, Halyomorpha halys (Hemiptera: Pentatomidae).

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 ±â€…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.

Evaluating deployment strategies for spotted lanternfly (Lycorma delicatula Hemiptera: Fulgoridae) traps.

The spotted lanternfly, Lycorma delicatula (White) (Hemiptera: Fulgoridae), is an invasive planthopper that was first detected in the United States in Berks County, PA, in 2014, and has since spread to 13 states in the Eastern United States. This phloem-feeding pest has a broad host range, including economically important crops such as grapevine, Vitis spp. Monitoring presence and relative abundance of L. delicatula is essential to develop pest management tools. Here, we compared deployment strategies to optimize use of L. delicatula monitoring traps. Standard circle traps, sticky bands, and circle traps with replaceable bag tops were deployed at sites with either high or low populations present. Trap deployment at different heights and on different host tree species and trap sampling intervals were evaluated for standard circle traps only. Circle traps captured significantly more L. delicatula adults at low-density sites compared with other trap types in 2021, and no differences were detected at high-density sights. Traps deployed 1 m from the ground captured significantly more adults than those deployed at 0.5 m; no differences were detected for nymphs. While no significant differences in captures were found among intervals, weekly or biweekly sampling prevented specimen degradation. Although traps deployed on Ailanthus altissima (Mill.) Swingle (Sapindales: Simaroubaceae) captured significantly or numerically more L. delicatula at most sites, traps deployed on other hosts also yielded consistent captures. We were also able to alter the construction of circle trap skirts to allow for deployment on different sized tree trunks.

Dendrochronology reveals different effects among host tree species from feeding by Lycorma delicatula (White).

The spotted lanternfly, Lycorma delicatula (White) (Hemiptera: Fulgoridae), was first detected in the United States in Berks County, Pennsylvania, in 2014. Native to China, this phloem-feeding planthopper threatens agricultural, ornamental, nursery, and timber industries in its invaded range through quarantine restrictions on shipments, as well as impacts on plants themselves. The long-term impacts of L. delicatula feeding on tree species have not been well studied in North America. Using standard dendrochronological methods on cores taken from trees with differing levels of L. delicatula infestation and systemic insecticidal control, we quantified the impact of L. delicatula feeding on the annual growth of four tree species in Pennsylvania: Ailanthus altissima, Juglans nigra, Liriodendron tulipifera, and Acer rubrum. The results suggest that L. delicatula feeding is associated with the diminished growth of A. altissima, but no change was observed in any other tree species tested. The results also suggest that systemic insecticides mitigate the impact of L. delicatula feeding on A. altissima growth.

Survival and development of Lycorma delicatula (Hemiptera: Fulgoridae) on common secondary host plants differ by life stage under controlled conditions.

Host range assessment for emerging invasive insects is a vital step toward fully defining the issues the insect may pose. Spotted lanternfly (SLF) is an invasive species that is rapidly expanding its presence in the United States. The primary hosts facilitating this spread are tree of heaven, a plant from SLF's native range, and the economically important winegrape. Black walnut is also implicated as an important and common host plant. This study investigated the survival and development of SLF on diets that included a variety of crop host plants in the presence or absence of tree of heaven. The following plant species, 'Honeycrisp' apple, 'Reliance' peach, silver maple, and tree of heaven were paired with winegrape or black walnut throughout the study. SLF had strong development and high survival on a diet of winegrape alone, and winegrape or black walnut paired with tree of heaven. Survival parameters were reduced with all other plant pairings. In particular, SLF in the winegrape and peach diet treatment did not develop past the third nymphal instar. A second experiment evaluated the survival of early and late instar nymphs and adult SLF life stages on three specialty crops - 'Cascade' hops, muscadine grapes, and kiwifruit over a two-week period. Nymphs survived longer than adults, with survival of first and second instar nymphs on hops not differing from the control tree of heaven treatment. The adult stage survived best on kiwi and muscadine grape. Our results show tree of heaven and winegrape were the only single plant diets evaluated that are sufficient for complete SLF development, while other host plants may require additional host or hosts of sufficient nutritional quality for SLF survival.

Evaluating invasion risk and population dynamics of the brown marmorated stink bug across the contiguous United States.

BACKGROUND: Invasive species threaten the productivity and stability of natural and managed ecosystems. Predicting the spread of invaders, which can aid in early mitigation efforts, is a major challenge, especially in the face of climate change. While ecological niche models are effective tools to assess habitat suitability for invaders, such models have rarely been created for invasive pest species with rapidly expanding ranges. Here, we leveraged a national monitoring effort from 543 sites over 3 years to assess factors mediating the occurrence and abundance of brown marmorated stink bug (BMSB, Halyomorpha halys), an invasive insect pest that has readily established throughout much of the United States. RESULTS: We used maximum entropy models to estimate the suitable habitat of BMSB under several climate scenarios, and generalized boosted models to assess environmental factors that regulated BMSB abundance. Our models captured BMSB distribution and abundance with high accuracy, and predicted a 70% increase in suitable habitat under future climate scenarios. However, environmental factors that mediated the geographical distribution of BMSB were different from those driving abundance. While BMSB occurrence was most affected by winter precipitation and proximity to populated areas, BMSB abundance was influenced most strongly by evapotranspiration and solar photoperiod. CONCLUSION: Our results suggest that linking models of establishment (occurrence) and population dynamics (abundance) offers a more effective way to forecast the spread and impact of BMSB and other invasive species than simply occurrence-based models, allowing for targeted mitigation efforts. Implications of distribution shifts under climate change are discussed. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Influence of Trap Location in the Tree Canopy on Captures of Adventive Trissolcus japonicus (Hymenoptera: Scelionidae).

Trissolcus japonicus (Ashmead) (Hymenoptera: Scelionidae) is an egg parasitoid of the invasive Asian pest, brown marmorated stink bug, Halyomorpha halys (Stål) (Hemiptera: Pentatomidae). Also native to Asia, adventive T. japonicus populations have been detected in North America since 2014, and are currently reported from 15 U.S. states, the District of Columbia, and two Canadian provinces. Yellow sticky cards (YSC) have proven effective for monitoring the presence, seasonal abundance, and distribution of these adventive populations. Our research has utilized YSC deployed in the midcanopy of H. halys host trees, following a study in which all leaves on felled tree of heaven, Ailanthus altissima (Mill.) Swingle, were inspected for H. halys egg masses, yielding eggs parasitized by T. japonicus only from mid- and upper-canopy leaves. However, given that other investigators have captured T. japonicus using YSC deployed in the lower-canopy, and that the effect of YSC placement in trees on T. japonicus captures had not been examined, captures of T. japonicus on YSC in the mid- and lower-canopy of individual A. altissima were compared. Traps were replaced weekly for five weeks and assessed for scelionid species. In 2020 and 2021, T. japonicus represented ≥53% of all Scelionidae captured, and there was not a significant effect of YSC location in the canopy on its captures. Deploying YSC at either canopy height was effective for measuring the relative abundance of T. japonicus, but sampling from the lower canopy substantially improved the efficiency and convenience of T. japonicus surveillance.

Seasonal Distribution of Halyomorpha halys (Hemiptera: Pentatomidae) Captures in Woods-to-Orchard Pheromone Trap Transects in Virginia.

The invasive brown marmorated stink bug, Halyomorpha halys (Stål), is a pest of numerous economically important crops in the USA. In the Mid-Atlantic region, it is a significant, direct pest in tree fruit orchards, many of which are bordered by woodlots containing a variety of its deciduous tree and shrub hosts. During the growing season, H. halys moves from woodland habitats into crops, but seasonal changes in its relative abundance between these adjacent habitats have not been examined. Using linear transects of six pheromone-baited pyramid traps that extended from 100 m into the interior of woodlots to 100 m into the interior of adjacent commercial apple orchards in Virginia, spatiotemporal changes in H. halys captures were measured during three growing seasons. Captures of H. halys adults and nymphs were recorded weekly from May through October, and annual data were separated into early, mid, and late-season captures. Only adults were captured during the early season, and there was no indication of a spatial trend in captures across traps in the transects among years. Beginning in mid-season and becoming increasingly apparent by late season, captures of H. halys adults and nymphs tended to become most frequent in traps at the woods and orchard edges and at 50 m into the orchard interior. These findings conform with and expand upon previous research documenting an edge effect for H. halys relative abundance and can inform and support the optimization of perimeter-based management strategies for H. halys in Mid-Atlantic apple orchards.

Survivorship and Development of the Invasive Lycorma delicatula (Hemiptera: Fulgoridae) on Wild and Cultivated Temperate Host Plants.

The invasive spotted lanternfly, Lycorma delicatula, (White Hemiptera: Fulgoridae) continues to spread throughout the Eastern United States. This species exhibits a broad host range, with tree of heaven, Ailanthus altissima (Mill.) Swingle, commonly referred to as the preferred host. Here, we evaluated 2-wk survivorship of early nymphal instars, late nymphal instars, and adult L. delicatula on single diets of ten wild and cultivated hosts: tree of heaven; apple, Malus domestica; peach, Prunus persica; black cherry, P. serotina Ehrh; black locust, Robinia pseudoacacia L.; black walnut, Juglans nigra L.; common hackberry Celtis occidentalis L.; mulberry Morus alba L.; sugar maple Acer saccharum Marshall; white oak, Quercus alba L.. Among them, early and late instars had significantly greater survivorship on tree of heaven and black walnut and adults had greatest survivorship on tree of heaven. Additionally, we evaluated development and survivorship of L. delicatula from newly hatched nymphs to adulthood on single diets of tree of heaven, black walnut, grapevine, apple, and peach, and mixed diets of tree of heaven plus one other host. Single host diets that supported L. delicatula development to adulthood were tree of heaven and black walnut. Interestingly, mixed diets also supported development, and reduced development time to adults by up to 12% compared with the single tree of heaven diet. Our results suggest that within agroecosystems and across landscapes, L. delicatula can develop on single hosts such as tree of heaven, but also on multiple host plants, yielding adults earlier in the growing season.

Development of rearing methodology for the invasive Spotted Lanternfly, Lycorma delicatula (Hemiptera: Fulgoridae).

Lycorma delicatula, White (Hemiptera: Fulgoridae), spotted lanternfly, is a univoltine, phloem-feeding, polyphagous and invasive insect in the USA. Although a primary host for this species is Ailanthus altissima, tree of heaven, L. delicatula also feeds on a wide range of hosts important to the USA including cultivated grapevines. Due to the need for classical or augmentative biological control programs to reduce impacts of L. delicatula across invaded areas, we developed a laboratory-based rearing protocol for this invasive species. Here, we evaluated the use of A. altissima apical meristems, epicormic shoots, and fresh foliage cut from A. altissima as a food source for rearing newly hatched L. delicatula. On these sources of plant material <20% of L. delicatula developed into adults and no oviposition occurred. However, when young, potted A. altissima trees were used as a food source, >50% of L. delicatula nymphs developed to the adult stage under natural daylengths and temperatures ranging from 20-25°C. The addition of wild grapevine, Vitis riparia, did not increase survivorship or reduce development time. To elicit mating and oviposition, adults were provided with A. altissima logs as an oviposition substrate and maintained under shortened daylengths and reduced nighttime temperatures (12L:12D and 24°C:13°C). This resulted in 2.12 egg masses deposited per female, which was 4× more than when adults were maintained in standard rearing conditions (16L:8D and 25°C). Based on these experiments, we present a protocol for reliably rearing L. delicatula under laboratory and/or greenhouse conditions.

Behavioral Effects and Retention of Protein Immunomarkers on Plum Curculio Conotrachelus nenuphar (Coleoptera: Curculionidae).

Protein immunomarking can be used to track the dispersal of insects in the field or identify plant-insect interactions. By marking insects with known proteins and recapturing them, their movement or host use can be quantified with Enzyme Linked Immunosorbent Assay (ELISA). Before using this technique, retention and behavioral effects of these markers should be evaluated to ensure that the insect's natural behaviors are conserved. Here, we tested the effects of protein markers on the plum curculio, Conotrachelus nenuphar (Coleoptera: Curculionidae) using two different application methods. This weevil is native to North American and a pest of tree fruit and blueberry in the United States and causes damage resulting in near complete crop loss if left untreated. We tested the effects of marking adult C. nenuphar with two inexpensive food-based immunoprotein markers, bovine casein (cow's milk) and chicken albumin (egg whites) on climbing distance (total cm), lateral movement (total cm), and lateral movement speed (cm/s), as well as retention time of protein immunomarkers. Neither protein immunomarker affected C. nenuphar movement or climbing, although females climbed significantly greater distances than males. ELISA assays detected 37.5-56.2% of milk protein and 56.2-59.3% of egg on the insect 7 d after application depending on application method. Our findings indicate that food-based protein immunomarkers can be used in future studies to test C. nenuphar movement within host plants without impacting behavior. The use of protein immunomarking will allow studies that will lead to behaviorally based management tactics.

Phenology of Lycorma delicatula (Hemiptera: Fulgoridae) in Virginia, USA.

The spotted lanternfly, Lycorma delicatula (White), is an invasive planthopper that was first discovered in North America in Berks County, Pennsylvania in 2014. Currently, L. delicatula has spread to eight additional states and threatens agricultural, ornamental, and timber commodities throughout the United States. The timing of insect life events is very important in the development of pest management tools and strategies. In 2019 and 2020, L. delicatula phenology was successfully documented in Winchester, Virginia using weekly 5-min observational surveys at established monitoring plots. Each year, L. delicatula were active in the environment from May to November with initial detections of first, second, third, fourth, and adults occurring in May, May, June, June, and July, respectively. Cumulative average growing degree days were also calculated for the onset of each L. delicatula life stage using local weather data and a lower developmental threshold of 10°C. First-instar L. delicatula were initially observed at 135 and 111.5, adults at 835 and 887, and egg masses at 1673.5 and 1611.5 in 2019 and 2020, respectively. Combined, these data can be used by growers and land managers to facilitate timing of effective pest management strategies.