Neonicotinoid insecticides are now the most widely used insecticides in the world. Previous studies have indicated that sublethal doses of neonicotinoids impair learning, memory capacity, foraging, and immunocompetence in honey bees (Apis mellifera, Linnaeus) (Hymenoptera: Apidae). Despite these, few studies have been carried out on the molecular effects of neonicotinoids. In this study, we focus on the second-generation neonicotinoid thiamethoxam, which is currently widely used in agriculture to protect crops. Using high-throughput RNA-Seq, we investigated the transcriptome profile of honey bees after subchronic exposure to 10 ppb thiamethoxam over 10 d. In total, 609 differentially expressed genes (DEGs) were identified, of which 225 were upregulated and 384 were downregulated. Several genes, including vitellogenin, CSP3, defensin-1, Mrjp1, and Cyp6as5 were selected and further validated using real-time quantitative polymerase chain reaction assays. The functions of some DEGs were identified, and Gene Ontology–enrichment analysis showed that the enriched DEGs were mainly linked to metabolism, biosynthesis, and translation. Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that thiamethoxam affected biological processes including ribosomes, the oxidative phosphorylation pathway, tyrosine metabolism pathway, pentose and glucuronate interconversions, and drug metabolism. Overall, our results provide a basis for understanding the molecular mechanisms of the complex interactions between neonicotinoid insecticides and honey bees.

The foraging behavior of different bee species is a key factor influencing the pollination efficiency of different crops. Most pear species exhibit full self-incompatibility and thus depend entirely on cross-pollination. However, as little is known about the pear visitation preferences of native Apis cerana (Fabricius; Hymenoptera: Apidae) and adventive Apis mellifera (L.; Hymenoptera: Apidae) in China. A comparative analysis was performed to explore the pear-foraging differences of these species under the natural conditions of pear growing areas. The results show significant variability in the pollen-gathering tendency of these honey bees. Compared to A. mellifera, A. cerana begins foraging at an earlier time of day and gathers a larger amount of pollen in the morning. Based on pollen collection data, A. mellifera shows variable preferences: vigorously foraging on pear on the first day of observation but collecting pollen from non-target floral resources on other experimental days. Conversely, A. cerana persists in pear pollen collection, without shifting preference to other competitive flowers. Therefore, A. cerana outperforms adventive A. mellifera with regard to pear pollen collection under natural conditions, which may lead to increased pear pollination. This study supports arguments in favor of further multiplication and maintenance of A. cerana for pear and other native crop pollination. Moreover, it is essential to develop alternative pollination management techniques to utilize A. mellifera for pear pollination.

Bemisia tabaci (Gennadius; Hempitera: Aleyrodidae) is considered to be a cryptic (sibling) species complex, the members of which exhibit morphological invariability while being genetically and behaviorally distinct. Members of the complex are agricultural pests that cause direct damage by feeding on plants, and indirectly by transmitting viruses that cause diseases leading to reduced crop yield and quality. In Pakistan, cotton leaf curl disease, caused by multiple begomovirus species, is the most economically important viral disease of cotton. In the study outlined here, the diversity and geographic distribution of B. tabaci cryptic species was investigated by analyzing a taxonomically informative fragment of the mitochondrial cytochrome c oxidase 1 gene (mtCOI-3′). The mtCOI-3′ sequence was determined for 285 adult whiteflies and found to represent six cryptic species, the most numerous being Asia II-1 and Middle East Asia Minor 1 (MEAM-1), the later also referred to as the B-biotype, which was previously thought to be confined to Sindh province but herein, was also found to be present in the Punjab province.The endemic Asia I was restricted to Sindh province, while an individual in the Asia II-8 was identified in Pakistan for the first time. Also for the first time, samples were collected from northwestern Pakistan and Asia II-1 was identified. Results indicate that in Pakistan the overall diversity of B. tabaci cryptic species is high and, based on comparisons with findings from previous studies, the distribution is dynamic.

Onion thrips, Thrips tabaci Lindeman, is a primary insect pest of onions (Allium cepa) worldwide. Onion thrips cause feeding damage by destroying epidermal tissue. They are also vectors of Pantoea ananatis (Serrano) Mergaert, the bacteria that causes center rot. Onions with center rot develop white streaks with water-soaked margins along the onion leaves, which turn necrotic and lead to bulb rot during storage.The role of thrips feeding on the establishment and progression of bacterial infection in onions has not been investigated. Onions infested with thrips and inoculated with P. ananatis had more necrotic tissue and symptoms were more severe with increasing thrips density. We conducted a fluorescence microscopy study that examined how P. ananatis (expressing a fluorescence protein gene) colonized a control group of onions without thrips in comparison to a test group of onions with thrips. We found that P. ananatis colonized some onions in the control group because of naturally existing wounds in the epidermal tissue but more colonization was found in the thrips infested group because of the increased presence of entry points caused by thrips feeding. Overall, our results demonstrate that wounds caused by thrips feeding facilitate center rot development by providing entry sites for the bacteria into leaf tissue.

Drosophila suzukii (Matsumura; Diptera: Drosophilidae) is an invasive vinegar fly and pest of soft fruits in North America, including wild blueberries (Vaccinium angustifolium Aiton) in Maine. Despite its presence in the continental United States for 9 yr, little is known about its natural enemy complex. Here we report the results of a 3-yr study designed to identify naturally-occurring predators in Maine's wild blueberry fields. Experiments were conducted to determine pupation site and pupation depth to understand D. suzukii's predation vulnerability. Predation rates in the field of fully-exposed, caged, and buried pupae were measured. Pitfall traps were deployed to identify the potential predator assemblage, and laboratory experiments were conducted to determine how many pupae were consumed by commonly occurring ground beetle species (Carabidae) and field crickets (Gryllus pennsylvanicus Burmeister).The most commonly collected predators were ants, ground beetles, harvestmen, and field crickets. Significantly more pupae were found to occur in the soil compared to blueberry fruit, with most pupae in the top 0.5 cm layer of soil. Pupal predation rates in the field were high, with higher rates of predation on exposed pupae compared to buried pupae. Laboratory studies revealed that ground beetles and field crickets are likely predators of D. suzukii pupae.

The purpose was to evaluate the side effects of strains Metarhizium anisopliae (Metsch.) Sorokin sensu lato Unioeste 43 and M. anisopliae sensu stricto ESALQ 1641 on Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) under controlled conditions. A free-choice test for parasitism was performed, with the confinement of T. pretiosum females mated with cards (1 × 5 cm) containing age-standardized Anagasta kuehniella Zeller eggs, either sprayed with a fungal strain (10⁹ conidia/ml) or 0.01% v/v Tween 80 (control). For the no-choice tests, T. pretiosum females mated were confined with cards sprayed with fungal strains before or after parasitism, and cards with fungal applications at different times. The number of parasitized eggs, percentage of emergence, longevity, egg-to-adult period, sex ratio, total and confirmed mortality by the fungus, and longevity of females that parasitized previously sprayed eggs, were assessed. Histological analysis of immature phases was also performed. The fungus was repellent to T. pretiosum in the free-choice test, while in the no-choice test, fungal applications before and after parasitism did not affect the number of eggs parasitized by T. pretiosum or the sex ratio of emerging adults. However, both strains affected adult emergence rates, the egg-to-adult period, and longevity. Overall, both M. anisopliae strains had minor effects on these biological parameters of T. pretiosum under controlled conditions. Hyphae were not detected in histological observations of immature stages of the parasitoid.

The sterile insect technique uses males that have been mass-reared in a controlled environment. The insects, once released in the field, must compete to mate. However, the mass-rearing condition supposes a loss of fitness that will be noticeable by wild females. To compare the fitness of wild males and mass-reared males, three competition settings were established. In setting 1, wild males, mass-reared males and wild females were released in field cages. In setting 2, wild females and wild males were released without competition, and in setting 3, mass-reared males and mass-reared females were also released without competition. Male fitness was based on their mating success, fecundity, weight and longevity. The fitness of the females was measured based on weight and several demographic parameters. The highest percentage of mating was between wild males and wild females between 0800 and 0900 h in the competition condition, while the mass-reared males started one hour later. The successful wild males weighed more and showed longer mating times, greater longevity and a higher number of matings than the mass-reared males. Although the mass-reared males showed the lowest percentage of matings, their fecundity when mating with wild females indicated a high fitness. Since the survival and fecundity of wild females that mated with mass-reared males decreased to become similar to those of mass-reared females that mated with mass-reared males, females seem to be influenced by the type of male (wild or mass-reared).

The effect of nine constant temperatures on developmental time of Oligota flavicornis (Boisduval and Lacordaire) preying on Tetranychus cinnabarinus (Boisduval) (Acari: Tetranychidae) eggs was determined under laboratory conditions of 75 ± 5% RH and a 16:8 (L:D) h photoperiod. O. flavicornis survival rates were highest between 18 and 30°C, although O. flavicornis eggs developed successfully to adults at 12–32°C, and the developmental durations to adult at the seven temperatures (12, 15, 18, 20, 25, 30, and 32°C) were 114.41, 51.66, 33.45, 23.21, 13.43, 11.54, and 17.18 d, respectively. Two linear and seven nonlinear models (Logan-6 and Logan-10, Taylor, Lactin-1 and Lactin-2, and Brière-1 and Brière-2) were fit to the developmental rates of the immature predatory stages to estimate the thermal constant (K) and critical temperatures.The lower temperature threshold (T₀) and K for the immature stages using the common linear model were 9.96°C and 225.73 degree-days and Ikemoto–Takai linear model were 11.01°C and 167.14 degree-days, respectively.The upper temperature threshold values estimated by the Logan-6 and Lactin-1 models were both 34.86°C.The T₀ values estimated by the Brière-1 and Brière-2 models were 10.67 and 9.32°C for all immature stages, respectively, and the estimated optimal temperature according to the Brière-2 model was 29.59°C. Therefore, the two linear models and Brière-2 model estimates approximated the actual relationship between the temperature and developmental rate of immature O. flavicornis.

The common green lacewing, Chrysoperla carnea (Stephens) (Neuroptera: Chrysopidae), is a well-known biocontrol agent.The current study examined host plant–herbivore-predator interactions with C. carnea and Myzus persicae on four host plants (peach, almond, pepper, and potato).The experiments were carried out at 25 ± 1°C and 65 ± 5% RH at a photoperiod of 16:8 (L:D) h). Duration of the preadult growth period, adult longevity, fecundity, and population growth parameters were analyzed based on the age-stage, two-sex life table theory. The shortest and longest preadult developmental times of the predator were observed on the peach and potato, respectively. The highest and lowest predation rate, oviposition period, and male and female longevity of predator were also observed on the peach and potato, respectively. The lowest intrinsic rate of increase (r) and finite rate of increase (λ) were observed on the potato (0.1087 and 1.11 d^–1, respectively) and the highest on the peach (0.1460 and 1.15 d^–1, respectively). The maximum and minimum mean generation times (T) were 41.84 and 35.59 d in the potato and peach, respectively. Overall, peach was found to be a more appropriate host than the other host plants for development and predation fitness of C. carnea. These findings reveal that information on tritrophic interactions and subsequent life table evaluation of natural enemies improves integrated pest management programs.

The kudzu bug, Megacopta cribraria (Fabricius), is an invasive pest of soybeans, Glycine max (L.) Merr., that has recently been detected in the United States.This study investigated whether there was a differential attraction of adult bugs to soybean growth stages, and whether the attraction was related to soybean constitutive volatiles. Greenhouse choice assays examined the behavioral orientation preference of adult bugs exposed to four growth stages of whole soybean plants: vegetative (V2), flowering (R1), pod (R3), and seed (R5). Results show that significantly more adults landed on plants in the early reproductive stage R1 than in other stages. Laboratory olfactometer assays also demonstrate that significantly more adult bugs were attracted to R1 plants, with females responding more strongly than males. Both greenhouse and olfactometer assays indicate that the differential attraction of adult bugs to soybean growth stages was mediated by plant constitutive volatiles.These results offer an insight into kudzu bug chemical and behavioral ecology and thus are of great significance for optimizing the timing of field scouting and treatment as well as the development of soybean pest management programs.

Plant volatiles help herbivores to locate their hosts, and therefore, they could be used to help develop pesticide-free pest management strategies.To develop an attractant for tea leafhopper (Empoasca onukii), we screened nine tea plant volatile compounds for their attractiveness using Y-tube olfactometer assays. Results indicated that tea leafhoppers significantly preferred ocimene, limonene, (Z)-3-hexenol, and (Z)-3-hexenyl acetate over clean air. These compounds were combined in a blend which lost its attractiveness at concentrations below 10^–2 g/ml in liquid paraffin. In field tests, the blend was attractive to leafhoppers only in autumn, but not in summer. Analyses of the tea field background odor showed that all four components of the blend were present at much higher concentrations in summer (0.05–0.001 ng/liter) than in autumn (∼10- to 25-fold lower). In fieldY-tube bioassays, compared with the tea field background odor, the blend was attractive at a concentration of 10^–1 g/ml in liquid paraffin, but not at 10^–2 g/ ml.These results suggest that field background odor can disrupt the attractiveness of an attractant based on plant volatiles to herbivores.

The sex pheromones of Ectropis grisescensWarren and Ectropis obliqua Prout were both reported to contain (Z,Z,Z)-3,6,9-octadecatriene (Z3,Z6,Z9-18:H) and (Z,Z)-3,9-cis-6,7-epoxy-octadecadiene (Z3,epo6,Z9-18:H). To clarify how these two sibling geometrids maintain premating isolation, the female sex pheromones of the two species were reexamined. Gas chromatography–electroantennographic detection (GC-EAD) and gas chromatography–mass spectrometry revealed two GC-EAD-active compounds, Z3,Z6,Z9-18:H and Z3,epo6,Z9-18:H, in E. grisescens female pheromone glands as well as an additional GC-EAD-active compound, (Z,Z)-3,9-cis-6,7-epoxy-nonadecadiene (Z3,epo6,Z9-19:H), in E. obliqua female pheromone glands. Synthesized Z3,Z6,Z9-18:H and Z3,epo6,Z9-18:H elicited dose-dependent electroantennogram (EAG) responses from male antennae of both E. grisescens and E. obliqua. However, Z3,epo6,Z9-19:H only elicited dose-dependent EAG responses from E. obliqua and limited EAG responses from E. grisescens at all doses. In wind-tunnel studies, lures that contained Z3,Z6,Z9-18:H and Z3,epo6,Z9-18:H attracted E. grisescens males and had no effect on E. obliqua males.The addition of Z3,epo6,Z9-19:H to the blend of Z3,Z6,Z9-18:H and Z3,epo6,Z9-18:H strongly attracted E. obliqua males but had a limited attraction for E. grisescens males. Thus, Z3,Z6,Z9-18:H and Z3,epo6,Z9-18:H were sex pheromone components of E. grisescens, whereas Z3,Z6,Z9-18:H, Z3,epo6,Z9-18:H and Z3,epo6,Z9-19:H were sex pheromone components of E. obliqua.The presence or absence of Z3,epo6,Z9-19:H played a central role in the premating isolation of these two sibling species.

Bradysia odoriphaga Yang et Zhang, the Chinese chive root maggot, is a devastating pest of agricultural plants causing significant losses in vegetable crops, edible mushrooms, and humus. To determine the effects of feeding on different host plants and diets on the life parameters of B. odoriphaga and its tolerance to stressful conditions, we analyzed the life-table data of B. odoriphaga reared on Chinese chive (Allium hookeri), Welsh onion (Allium fistulosum), garlic (Allium sativum), oyster mushroom (Pleurotus ostreatus), and humus and assayed its tolerance to heat and insecticides. Furthermore, we assayed the physiological responses of B. odoriphaga reared on different hosts. Development from egg to adult was successfully completed on five hosts. Life table indicated that when reared on Chinese chive and oyster mushroom, B. odoriphaga showed better life parameters (shorter development time, higher survival rate, and fecundity) than on humus and garlic. However, B. odoriphaga larvae fed on humus and garlic better tolerated heat and insecticides than those that were fed Chinese chive and oyster mushroom; larvae survived longer at 38 and 40°C heat shock and at higher insecticide doses. Activities of antioxidant and detoxification enzymes varied significantly in larvae fed on different hosts. Catalase, superoxide dismutase, glutathione S-transferase, and carboxylesterase activities were higher in larva reared on garlic and humus than on Chinese chive and oyster mushroom. These findings indicated that B. odoriphaga preferred liliaceous vegetables and mushroom, and their tolerance against stress increased when reared on humus and garlic. Such basic information can promote targeted pest management in different agricultural fields and allow better understanding of the acclimatization strategy of B. odoriphaga.

Reproductive plasticity is a key determinant of species invasiveness. However, there are a limited number of studies addressing this issue in exotic insects.The rice water weevil Lissorhoptrus oryzophilus Kuschel (Coleoptera: Curculionidae), which is native to North America, is one of the most invasive insect pests in east Asia. In this study, we investigated the reproductive status of first-generation females (progeny of overwintered weevils) from five geographic regions in southern and northern China in the field, and reproductive status and ovipositional features of females provided with suitable host plants in the laboratory after collection. Under field conditions, a proportion of females oviposited, while the rest remained in diapause from all three southern regions investigated, but reproductive development did not take place in females from the two northern regions, where the weevil produces only one generation per year. However, when fed host plants in the laboratory, females from all regions laid eggs.They typically had a very short ovipositional period (3–6 d on average on rice at a temperature of 27°C), laid a low number of eggs, and did not die soon after oviposition; this was different from common reproductive females. We concluded that first-generation L. oryzophilus females, which largely enter diapause after emergence, are highly plastic in their reproductive performance and are ready to reproduce under favorable conditions. Our results indicate the significance of their reproductive plasticity for geographic spread and population development.

Large numbers of flies are needed to produce the quantity of larvae required for insect bioconversion of waste. However, this ‘mass-rearing’ may negatively affect adult survival and reproductive output.This study assessed the suitability for mass-rearing of four blow fly species, Chrysomya chloropyga, Chrysomya chloropyga (Wiedemann), Chrysomya megacephala (F.), Chrysomya putoria (Wiedemann) and Lucilia sericata (Meigen). Flies were kept at densities of 20, 50, 100, 250, 500, and 1,000 flies per 30 × 30 × 30 cm cage with an even sex ratio. Time to 50% mortality (LT50) was recorded, and the effects of density, species, and sex on LT50, fecundity, and fertility were determined. Females survived longer than males across all species. There was evidence for a trade-off between survival and high fecundity in L. sericata and C. chloropyga at density 250. C. megacephala had low fecundity across all densities. At high densities, C. putoria had the lowest mortality and highest fecundity, making it the most suitable for mass-rearing.

Tephritid mass-rearing systems require an artificial substrate for pupation. Pupation substrate characteristics influence the quality of insects produced. Coconut fiber, as an alternative to the conventional pupation substrate vermiculite, was evaluated for Anastrepha obliqua Macquart (Diptera: Tephritidae) pupation behavior (pupation patterns, distribution, respiration rate, and pupal weight) and adult fitness (adult eclosion time, flight ability, and male mating competitiveness). Pupation percentage at 24 h, pupal weight, and flight ability were not significantly affected by substrate type. Adult eclosion levels of 50% were reached at 29.7 and 41.6 h for coconut fiber and vermiculite, respectively. Pupae distribution patterns differed between substrates because the larval aggregation level was reduced during the pupation process in coconut fiber. The pupae aggregation was three times greater in vermiculite than in coconut fiber. A higher respiratory rate in the last days of pupation and adult eclosion were recorded in the insects maintained in coconut fiber. Coconut fiber suitability as a pupation substrate for quality mass production of pupae and its implications for sterile insect technique are discussed.

Production of feces (PF) is a useful proxy indicating quantity of ingested food. Although influenced by many uncontrolled factors PF provides insight into food consumption under natural conditions. In Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) variation in PF was investigated in populations of various hostplants at localities of central Bohemia (50N, 14E), throughout the season of 2016. The adults collected from these hostplants were starved for 48 h at standard conditions, and dry mass of feces produced during this period was measured. Despite enormous differences in PF among individuals, significant variation over the season occurred in average PF of both males and females. PF increased with abundance of aphids and was significantly greater in females than males. Gravidity, as manifested through oviposition within 48 h after capture, was associated with increased PF, while hostplant and color morph did not affect variation in PF among individuals. From PF as measured in this study, it can be estimated that at sites hosting abundant aphid populations H. axyridis (as an adult male or female) may consume 19 (male) to 45 (female) aphids per day (assumed body length 1mm). In the absence of aphids, adults may consume one to nine individuals of alternative prey per day (body length 1–2 mm).

Inspite of well-established potentiality of plant essential oils as biopesticides, their environmentally low persistence is considered as a hindering obstacle for its commercialization. In the present study, chemical composition and toxicity of essential oils isolated from leaves of Thymus eriocalyx and Thymus kotschyanus were evaluated against two-spotted spider mite, Tetranychus urticae. The chemicals present in the crude oil were found to be thymol (28.83%), oleic acid (11.51%), palmitic acid (8.60%), borneol (5.72%), ρ-cymene (3.60%), and 1,8-cineole (3.57%) in the essential oil of T. eriocalyx, and camphene (35.59%), linalyl acetate (20.47%), linalool (14.75%), α-terpineol (13.87%), and geranyl acetate (3.07%) in the essential oil of T. kotschyanus.The essential oils had strong fumigant toxicity on the adult females of Te. urticae and their fumigation persistence was prolonged until 6 and 5 d, respectively, for T. eriocalyx and T. kotschyanus. Loading of essential oils in MCM-41 increased their stability and persistence was extended up to 20 and 18 d for T. eriocalyx and T. kotschyanus. Further, mite mortality increased from 80 to 203 mites by T. eriocalyx and from 58 to 186 mites by T. kotschyanus nanoencapsulated essential oils. Based on these results, nanoencapsulation of T. eriocalyx and T. kotschyanus essential oils in MCM-41 may be a useful method for their application in the management of Te. urticae.

The coffee berry borer (CBB), Hypothenemus hampei Ferrari (Coleoptera: Curculionidae: Scolytinae) is a recent invader to Hawaii.To date, limited information regarding the seasonal phenology of this pest on the islands limits the implementation of integrated control strategies. As part of a coffee farmer training program, we monitored CBB flight activity in 15 coffee plantations (Kona and Kau Districts) over 10 mo with methanol-ethanol (3:1 ratio) baited traps. Concurrently, we quantified CBB infestation and penetration rates inside developing coffee berries through the end of harvest. Approximately 1 million CBB were captured, with the highest activity (e.g., >500 CBB/trap/wk) in December through February, coinciding with end of main regional harvesting periods. Relatively high activity (>250 CBB/trap/wk) was also observed during berry development, in May and June (Kona) and June and July (Kau). Field infestation rates were higher overall in Kau (9.6 ± 1.1%) compared with coffee plantations in Kona (4.7 ± 0.4%). Linear regression investigated relationships between CBB trap data and berry infestation rates. Trap catch data generally correlated better with the proportion of shallow entries (AB position) compared with deeper penetrations (CD position) or total infestation. Pearson correlation coefficients based on different parameters (i.e., region, altitude, and berry phenology) revealed positive and mostly significant correlations between these variables (R values 0.410 to 0.837). Timing peak flight activity of CBB with insecticide applications will help coffee growers improve pest control.The ability of trap data to calculate reliable economic (action) thresholds for the CBB is discussed.

Since the discovery of Aphis glycines Matsumura (Hemiptera: Aphididae) in the United States, the primary management tactic has been foliar insecticides. Alternative management options such as host plant resistance to A. glycines have been developed and their effectiveness proved. However, the use of host plant resistance was complicated by the discovery of multiple, virulent biotypes of A. glycines in the United States that are capable of overcoming single Rag genes, Rag1 and Rag2, as well as a two-gene pyramid of Rag1+Rag2. However, current models predict that the virulent allele frequency of A. glycines decreases in response to the use of pyramided Rag genes, suggesting that pyramids represent a more sustainable use of these traits. Previous research has demonstrated that virulent biotypes can be effectively managed using a three-gene pyramid of Rag1+Rag2+Rag3. Additional Rag-genes have been discovered (Rag4 and Rag5), but whether the incorporation of these genes into novel three-gene pyramids will improve efficacy is not known. We tested single-gene (Rag1 and Rag2) and pyramid cultivars (Rag1+Rag2, Rag1+Rag2+Rag3, Rag1+Rag2+Rag4) to multiple biotypes in laboratory assays. Our results confirm that the Rag1+Rag2+Rag3 pyramid effectively manages all known A. glycines biotypes when compared with cultivars that are overcome by the associated biotype. Our results indicate that Rag1+Rag2+Rag4 would be an effective management option for biotype-1, biotype-2, and biotype-3 A. glycines, but had a negligible impact on biotype-4.

Pest-induced changes in plant reflectance are crucial for the development of pest management programs using remote sensing. However, it is unknown if plant reflectance data is also affected by foliar insecticides applied for pest management. Our study assessed the effects of foliar insecticides on leaf reflectance of soybean. A 2-yr field trial and a greenhouse trial were conducted using randomized complete block and completely randomized designs, respectively. Treatments consisted of an untreated check, a new systemic insecticide (sulfoxaflor), and two representatives of the most common insecticide classes used for soybean pest management in the north-central United States (i.e., λ-cyhalothrin and chlorpyrifos). Insecticides were applied at labeled rates recommended for controlling soybean aphid; the primary insect pest in the north-central United States. Leaf-level reflectance was measured using ground-based spectroradiometers. Sulfoxaflor affected leaf reflectance at some red and blue wavelengths but had no effect at near-infrared or green wavelengths. Chlorpyrifos affected leaf reflectance at some green, red, and near-infrared wavelengths but had no effect at blue wavelengths. λ-cyhalothrin had the least effect on spectral reflectance among the insecticides, with changes to only a few near-infrared wavelengths. Our results showing immediate and delayed effects of foliar insecticides on soybean reflectance indicate that application of some insecticides may confound the use of remote sensing for detection of not only insects but also plant diseases, nutritional and water deficiencies, and other crop stressors.

This study was conducted to develop temperature-dependent postdiapause development models of overwintering larvae of the Asian corn borer, Ostrinia furnacalis Guenée (Lepidoptera: Crambidae), and to evaluate the models' forecasting accuracy using spring adult emergence data. Overwintering larvae were collected at three different times: 24 February (first), 23 March (second), and 25 April (third), 2005.The developmental periods of each collection colony were measured at eight constant temperatures, and those developmental rates were modeled with linear and nonlinear regression. One linear and three nonlinear models provided good fits of developmental rate to temperature across all colonies (r² = 0.96–0.99). The distribution of development completion time was modeled with a Weibull equation that fit data from the second (r² = 0.92) and third (r² = 0.97) colonies better than the first (r² = 0.87). A Lactin 2 model based on data from the first colony was statistically the best model to describe the relationship between temperature and the postdiapause development rate of O. furnacalis (r²_adj = 0.99). However, validation results based on the field data showed that the Logan 6 model combined with the Weibull model (based on the second colony) was well describing spring adult emergence patterns up to 50% cumulative emergence date.

Hydrolyzed protein lures are widely used to monitor fruit fly pests but are rapidly degraded by microbial activity and must be replaced frequently. To improve the stability of lures, the quaternary ammonium biocide, benzalkonium chloride (BC), was evaluated in mixtures with two hydrolyzed proteins commonly used to monitor Anastrepha spp. The mean number of Anastrepha obliqua adults captured during six consecutive weeks using Captor + borax with the addition of 240 mg BC/liter, not renewed during the test, was similar to Captor + borax that was replaced at weekly intervals and was more effective than Captor + borax without BC. Numbers of A. obliqua flies captured in 30% CeraTrap diluted in water containing 240 mg BC/liter were similar to those caught in traps baited with Captor + borax or 30% CeraTrap without BC in the first 9 d of evaluation but was significantly more effective than both lures after 56 d. After >2 mo of use, 30% CeraTrap containing 240 mg BC/liter remained as effective as newly prepared 30% CeraTrap. The addition of BC to lures reduced surface tension of liquid lures by ∼40–50%. However, when BC was increased to 720 mg BC/liter, only a small additional reduction in surface tension was observed and higher concentrations of BC did not increase capture rates. These findings could contribute to reduced costs for trapping networks and the development of long-lasting formulations of liquid protein lures for bait stations and mass-trapping targeted at major tephritid pests.

Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), commonly called ‘Queensland fruit fly’ in Australia, and Mediterranean fruit fly (Ceratitis capitataWiedemann) (Diptera:Tephritidae) are the two most economically important fruit fly in Australia with B. tryoni in the east and Mediterranean fruit fly in the west. The two species coexisted for several decades, but it is believed that B. tryoni displaced Mediterranean fruit fly. In southeastern Australia, this was deemed inadequate for export market access, and a large fruit fly free zone (fruit fly exclusion zone) was developed in 1996 where B. tryoni was eradicated by each state department in their portion of the zone. This zone caused an artificial restricted distribution of B. tryoni. When the fruit fly exclusion zone was withdrawn in Victoria and New South Wales in 2013, B. tryoni became endemic once again in this area and the national distribution of B. tryoni changed. For export markets, B. tryoni is now deemed endemic to all eastern Australian states, except for the Greater Sunraysia Pest-Free Area. All regulatory controls have been removed between eastern states, except for some small zones, subject to domestic market access requirements. The eastern Australian states now form a B. tryoni endemic trading group or block. All Australian states and territories maintain legislation to regulate the movement of potentially infested host fruit into their states. In particular, eastern states remain active and regulate the entry of commodities possibly infested with Mediterranean fruit fly. The combination of regulatory controls limits the chances of Mediterranean fruit fly entering eastern states, and if it did, Mediterranean fruit fly is unlikely to establish in the opposition to a well-established B. tryoni population.

It has been suggested that fruit wastes including dropped and unharvested fruits, and fruit byproducts (i.e., pomace) found in fruit plantings and cideries or wine-making facilities could serve as potential off-season breeding sites for spotted wing Drosophila (Drosophila suzukii Matsumura (Diptera: Drosophilidae)). This idea, however, has yet to be widely tested. The goal of our study was to determine the potential of dropped fruit and fruit wastes as Fall spotted wing Drosophila breeding resources in Michigan, USA. Fruit waste samples were collected from 15 farms across the lower peninsula of Michigan and were evaluated for spotted wing Drosophila and other drosophilid emergence and used in host suitability bioassays. All of the dropped apples, pears, grapes, and raspberries and 40% of apple and 100% of grape fruit pomace evaluated were found to contain spotted wing Drosophila with the highest numbers collected from dropped grapes and pears. Greater spotted wing Drosophila recovery was found in fruit wastes at sites attached with cideries and wine-making facilities and with multiple cultivated fruit crops than sites with no cideries and only one crop. Females oviposited in raspberry, pear, apple, grape, apple pomace and grape pomace samples with the highest rates of reproduction in raspberries. Our results demonstrate that fruit wastes including dropped berry, pomme and stone fruits, as well as fruit compost may be important late season reproductive resources for spotted wing Drosophila.

Bagrada hilaris (Burmeister) (Hemiptera: Pentatomidae) is a serious pest of cruciferous crops in the central coast of California. Management of B. hilaris primarily involves the use of broad spectrum insecticides, such as pyrethroids and neonicotinoids, when the pest invades a crop field. Insect growth regulators (IGRs) are known for their efficacy on nymphal stages of B. hilaris, but little is known about their transovarial effects.Thus, the major objective of this study was to determine the transovarial effects of IGRs such as novaluron, diflubenzuron, and azadirachtin, when adults are exposed to direct topical spraying and dried residues. In addition, a direct topical spray of IGRs on older instars (fourth and fifth instars) was examined at 1-, 3-, and 7-d post-treatment.The number of young instars (first and second instars) and adults was recorded up to ∼31 d after exposure. In the topical spray assays, the number of young instars was significantly lower for novaluron, diflubenzuron, and azadirachtin than for the nontreated control.The number of young instars that emerged after novaluron treatment was low (0–11%). In the assays with dried residues, the number of young instars that developed was significantly lower for novaluron treatment than for the other IGRs and the nontreated control.There was no significant difference among diflubenzuron, azadirachtin, and nontreated control treatments in the development of young instars. In the assays with a topical spray of IGRs on older instars, significantly more nymphs died at 3 and 7 d after exposure.The IGRs had no clear impact on adults.

The southern green stink bug, Nezara viridula (L.) (Hemiptera: Pentatomidae), is a serious insect pest of tomatoes in Florida. In this study, we examined the use of three species of trap crops to manage N. viridula in North Florida tomato crops in 2014 and 2015. We used striped sunflower (Helianthus annuus) (Asterales: Asteraceae) and wild game feed sorghum (Sorghum bicolor) (Poales: Poaceae) in both years, but different species of millet each year: browntop millet (Panicum ramosum) (Poales: Poaceae) in 2014 and pearl millet (Pennisetum glaucum) (Poales: Poaceae) in 2015. The number of stink bug adults collected from wild game feed sorghum exceeded the number from sunflower, and none were collected from either species of millet. Sorghum attracted a significantly higher number of adults than did striped sunflower; however, both sunflower and sorghum attracted the adults of N. viridula. Adults of the pest feed on the sorghum panicle and sunflower head (inflorescence). Although fewer stink bugs were found feeding on sunflower, the sunflower was found to be a good source of other natural enemies and pollinators and also attracted significantly greater numbers of the brown stink bug Euschistus servus (Say) (Hemiptera: Pentatomidae) (another pest of tomatoes). While this study demonstrated the effectiveness of sorghum, we recommend that sorghum be planted with another trap crop, preferably sunflower, for better preventive control of the southern green stink bug.

Phthorimaea operculella Zeller has proven to be a limitation to potato production. Although pesticides can effectively reduce P. operculella populations, information regarding effective cultural controls is lacking. The following study was designed to evaluate the effect of different irrigation regimes and planting depths to reduce P. operculella damage under central pivot irrigation. Field trials were conducted in 2007 and 2008 and experimental plots followed standard growers' agronomic practices. Three levels of irrigation were used: optimal, 10% less than optimal, and 10% above optimal; potato planting depth was arranged as shallow, normal, and deep; normal planting was considered tubers planted at 0.15 m depth. Mines and number of larvae per plant were counted weekly until harvest and tuber damage, above and below ground, was estimated. Overall, irrigation had a stronger effect than planting depth but there were not a clear planting depth:irrigation interaction. Both years, sampling dates had a strong effect on the number of mines per plant where the number of mines per plant increased as season progressed.There were not clear trends related to mines per plant, mines per tuber or yield related to planting depth, but there were strong tendencies related to irrigation. The data presented herein provide valuable information regarding the potential to adjust irrigation to warrant better P. operculella control.

The common blossom thrips, Frankliniella schultzeiTrybom (Thysanoptera: Thripidae), is an important lettuce pest worldwide. Conventional sampling plans are the first step in implementing decision-making systems into integrated pest management programs. However, this tool is not available for F. schultzei infesting lettuce crops. Thus, the objective of this work was to develop a conventional sampling plan for F. schultzei in lettuce crops. Two sampling techniques (direct counting and leaf beating on a white plastic tray) were compared in crisphead, looseleaf, and Boston lettuce varieties before and during head formation. The frequency distributions of F. schultzei densities in lettuce crops were assessed, and the number of samples required to compose the sampling plan was determined. Leaf beating on a white plastic tray was the best sampling technique. F. schultzei densities obtained with this technique were fitted to the negative binomial distribution with a common aggregation parameter (common K = 0.3143). The developed sampling plan is composed of 91 samples per field and presents low errors in its estimates (up to 20%), fast execution time (up to 47 min), and low cost (up to US $1.67 per sampling area). This sampling plan can be used as a tool for integrated pest management in lettuce crops, assisting with reliable decision making in different lettuce varieties before and during head formation.

Abstract

Bagrada hilaris (Burmeister) (Hemiptera: Pentatomidae) is a non-native stink bug that feeds primarily on cole crops and wild mustards. Its invasion into desert agriculture in California and Arizona presents a conundrum between rapid pest development at warm temperatures and severe damage to cool season crops. In this study, the development and survival of B. hilaris were determined at nine constant temperatures (ranging from 20–42°C) when reared on organically grown broccoli florets. Egg hatching was greatly delayed at 20°C, and first instar nymphs did not survive at this temperature. No eggs hatched at 42°C. The highest survival rates (70.0–86.7%) of B. hilaris were observed at temperatures ranging from 24 to 35°C. The total developmental rate of B. hilaris from egg to adult increased from 0.027 to 0.066/d from 24 to 35°C, and then slightly dropped to 0.064/d at 39°C. Based on the linear model, B. hilaris requires 285.4 degree-days to complete its development. The Briere 1 model predicted the lower and upper temperature thresholds as 16.7 and 42.7°C, respectively.The optimal temperature for development (T_Opt) was estimated as 36°C. According to the results, B. hilaris is well adapted to warm conditions, and temperatures of 33–39°C are well suited for B. hilaris development. Information from this study helps explain the rapid range expansion of B. hilaris across the southern United States and will be instrumental in predicting future expansion across the rest of the country and in other parts of the world. The relationship between thermal thresholds and invasion dynamics of this pest are discussed.

Many female parasitoid wasps optimize host selection to balance the benefits of high-quality hosts and the costs of predator- or hyperparasitoid-induced mortality risks to maximize their fitness. Cannibalism exists in many insect species and affects survival of parasitoid larvae developing in or on parasitized hosts. However, little is known about how parasitoid wasps resolve the fitness consequence of host cannibalism-induced mortality risk during host selection. We examined the effect of oothecal age on cannibalism in the American cockroach Periplaneta americana (L.) (Dictyoptera: Blattidae) and its effect on host age selection and fitness of its oothecal parasitoid Evania appendigaster (L.) (Hymenoptera: Evaniidae). P. americana differentially cannibalized 1-d-old (30–60%) versus 10- to 40-d-old oothecae (<9%). However, parasitoid females did not avoid but still preferred to parasitize 1-d-old (45%) over 10- to 40-d-old oothecae (1.6–20%). The parasitism rate was greater and the handling time was shorter on 1-d-old compared to older oothecae. For parasitoid progeny emerging from different-aged oothecae, regression analysis showed that development time increased and body size (measured as hind tibia length) and longevity decreased with oothecal age. These results demonstrate that reduced parasitoid progeny survival due to host cannibalism did not change the parasitoid's oviposition preference for newly laid oothecae, and that E. appendigaster females traded progeny survival for fitness gains for themselves and their progeny.

Spatial assessment of molting in workers of Coptotermes formosanus Shiraki was conducted in laboratory-reared colonies using extended foraging arenas. Workers at a premolt stage were found concentrated in the nest or in a planar arena near the nest. However, molting individuals were found exclusively in the central nest and they stayed inside or near the central nest for at least 36 h postmolting.The absence of premolt workers at foraging sites suggests that the workers have an affinity to the nest for molting and the second study on nest-fidelity evaluation suggested that the workers molt in the proximity of eggs.The molting site fidelity by workers in a colony ensures that speeding up the time for mortality induced by chitin synthesis inhibitor (CSI) baits will not result in an inhibitory cascade of dead termites around the bait stations. Thus, speeding up the elimination of a C. formosanus colony using CSI baits with the addition of molt-accelerating compounds will not lead to secondary repellency. Reasons for the molting-site fidelity amongst workers in a colony are discussed.

German cockroaches (GCRs), Blattella germanica (L.) (Dictyoptera: Blattellidae), are attracted to those beer semiochemicals (e.g., ethanol) that formerly living and active yeasts have produced or otherwise formed in the brewing process. We predicted that an earlier step in the production of beer, where yeasts actively metabolize the sugar in malted barley powder (dry malt extract [DME]), is very attractive to GCRs. In laboratory experiments, a 3-component composition (3CC) comprising DME, water, and Brewer's yeast strongly attracted GCR nymphs, females, and males. Both Brewers' yeast and ‘spoilage organisms’ in the DME or water seem to add to the attractiveness of the 3CC, but there is no additive or synergistic effect between them.The 3CC becomes optimally attractive to GCRs after 12 h of fermentation and stays that attractive for at least 120 h. In field trapping experiments, the 3CC and—unexpectedly—also the DME each proved as effective for attracting and capturing GCRs as a commercial cockroach bait (Combat Roach Gel). Future studies will investigate lethal biocontrol agents that can be added to the 3CC, or the DME, and will explore the efficacy of such lethal baits for GCR control.

The aim of this study was to test the bioavailability and degradation in soil of the termiticides bifenthrin and fipronil, which are used to treat subterranean termites (Heterotermes indicola, Wasmann). Soil collected from different areas of Lahore was categorized as sandy clay loam (SCL) or sandy loam (SL). Laboratory bioassays were conducted to determine the bioavailability ratio of bifenthrin and fipronil in each type of soil after different periods of time. LT₅₀ values were determined posttreatment at different time intervals. Regarding soil type, both termiticides were more effective in SL soil, compared with SCL soil posttreatment. There were significant differences in termite mortality in treated compared with untreated control samples (P < 0.005). To test the degradation rate of soil termiticides, both qualitative and quantitative analyses were carried out by HPLC, and the effect of time on termiticide recovery rate determined. Calculated half-life values for bifenthrin (maximum, 1,002 and 1,262 d in SCL soil and SL soil, respectively) indicated that it persisted in both soil types at all concentrations. The maximum calculated half-life values of fipronil were 270 and 555 d in SCL and SL soil, respectively. At lower concentrations and over longer periods of time, fipronil completely degraded in SL soil, while a negligible amount was detected in SCL soil. Termiticide concentration decreased over time, as did the termiticide recovery rate. Overall, bifenthrin was more persistent than fipronil under all treatment conditions tested.

Termite baits using chitin synthesis inhibitors can protect structures from subterranean termite damage by eliminating colonies. One of the potential shortcomings of baits is that termites may have to feed extensively on a bait station for the colony to be eliminated. If disturbances occur during this critical feeding time, termites may abandon the site, which could prevent colony elimination. We exposed whole laboratory colonies of Coptotermes gestroi (Wasmann) containing an average of 59,000 termites to 0.5% noviflumuron baits, from 1 to 10 d, to simulate a temporary access to the bait as a result of disturbances. Our results show that colonies which fed for a single day and consumed as little as 1.1 g of bait matrix (=5.5 mg noviflumuron) were eliminated within 90 d. This study indicates that only a fraction of the bait matrix is necessary for elimination of large laboratory colonies, and that hypothetical disturbance reducing the access to a bait station would likely have little impact on the effect of the bait. Within 1 d, colonies had already acquired a lethal dose, which is shared by trophallaxis to all individuals of the colony, and colonies inevitably died despite short exposure to noviflumuron. It is estimated that if a field mature colony of 1 million individuals consumes more than 18.6 g of bait (=93 mg noviflumuron) in a relatively short period, it would already have acquired a colony-wide lethal dose and reached a point of no-return for colony elimination.

The house fly, Musca domestica L. (Diptera: Muscidae), is a major pest of medical and veterinary importance all over the world. Management efforts for house flies are usually compromised owing to their resistance to many groups of conventional insecticides. Cantharidin, a natural toxin produced by meloid beetles, is a biopesticide with a reported toxicity to some insect pests including house flies. However, the effects of cantharidin on biological and fitness parameters of house flies have not yet been investigated. In the present study, we investigated the toxicity and sublethal effects of cantharidin on biological parameters of house flies for two consecutive generations. The results revealed that the values of LC_50, LC₂₅, LC₁₀, and LC₂ against house flies were to be 2.45, 1.23, 0.66, and 0.30 mg/ liter, respectively. Sublethal effects of these concentrations on the development and reproduction parameters of house flies revealed that cantharidin reduced population growth by affecting pupation rate, adult emergence, and by lengthening developmental time. The female ratio, fecundity, egg hatching, and survival of adult flies were significantly reduced at LC_2, LC₁₀, LC₂₅, and LC₅₀ of cantharidin when compared with the control group. Furthermore, the increase in concentration of cantharidin had a significant effect on reducing the mean values of mean relative growth rate, net reproductive rate (R_o), intrinsic rate of natural increase (r_m), and biotic potential (bp). In conclusion, the results of this study revealed the toxicity of cantharidin against house flies and the adverse effects of sublethal concentrations on biological parameters which may have positive implications for effective management of house flies.

RNA interference is a powerful tool against corn rootworm. Adults and neonates of southern corn rootworm, Diabrotica undecimpunctata howardi Barber (Coleoptera: Chrysomelidae), were exposed to the LC₅₀ of vATPase-A and Snf7 double-stranded RNAs (dsRNAs), and the effects on female fecundity, egg viability, male fitness as measured by sperm viability and mating capacity, larval recovery along with dry weight, and instar determination 10 d after exposure to dsRNA, were determined. Significant reductions were observed for a number of parameters in dsRNA-exposed rootworms relative to control treatments. Female fecundity and larval recovery were significantly reduced after exposure to both dsRNAs. In addition, larval dry weight and recovery of 2nd and 3rd instars along with dry weight for 3rd instars were significantly reduced after neonate exposure to vATPase-A dsRNA. Neither dsRNA affected male capacity to mate or sperm viability after exposure to the respective LC₅₀s. After 10 d of feeding on untreated corn roots, neonates that survived exposure for 2 d to the vATPase-A dsRNA LC₅₀ exhibited lower dry weight than the control.There was significant gene knockdown in adult males and females after exposure for 5 d to LC₅₀ of vATPase-A and Snf7 dsRNAs.The parameters are discussed in terms of fitness and possible outcomes after deployment of corn hybrids expressing dsRNAs.

Chilo suppressalis (Walker; Lepidoptera: Crambidae) is one of the most damaging rice pests in China. Insecticides play a major role in its management. We describe how we monitored the resistance of C. suppressalis to four insecticides in seven field populations from Jiangxi, Hubei, and Hunan Provinces, China, in 2014–2016.The topical application method for resistance monitoring was suitable for triazophos, monosultap, and abamectin. The conventional rice seedling dipping method proved ineffective for testing chlorantraniliprole so the new artificial diet incorporation method was substituted. This new method provided more consistent results than the other methods, once baseline toxicity data had been established. All populations had moderate to high resistance to triazophos from 2014 to 2016. Monosultap resistance in two populations increased from low in 2014 to moderate in 2016 and the other five populations showed moderate to high-level resistance throughout. Abamectin resistance in three populations increased from sensitive or low in 2014 to moderate in 2015–2016, and the other populations had moderate to high levels of resistance. Resistance to chlorantraniliprole increased from sensitive or low in 2014 to moderate to high in 2016.These results suggested that resistance management strategies should be developed according to the needs of a specific location. It was suggested that, in these localities, organophosphate insecticides should be prohibited, the application of nereistoxin, macrolide antibiotic, and diamide insecticides should be limited, and other insecticides, including spinetoram and methoxyfenozide, that exhibited no resistance should be used rationally and in rotation to delay resistance development.

Mosquito control districts conduct rigorous insecticide treatments against both larval and adult Culex quinquefasciatus Say (Diptera: Culicidae), the primary vector of West Nile virus in the southern United States. However, the development of resistant populations of Cx. quinquefasciatus in response to extensive larvicide or adulticide applications has been demonstrated repeatedly across the world. Examining changes in insecticide susceptibility in treated field areas can help inform mosquito control districts as to whether or not their treatments remain effective. We hypothesized that frequent insecticide applications for the control of mosquitoes in East Baton Rouge Parish, Louisiana, lowered susceptibility of wild Cx. quinquefasciatus to larvicides. Larvicide susceptibility was measured using Lysinibacillus sphaericus, spinosad, and temephos in populations of Cx. quinquefasciatus sampled from sites in three Parishes where frequencies of insecticide applications varied, and frequencies of resistance were measured relative to a susceptible reference colony. Susceptibility to these larvicides was widespread, although fourfold resistance to the organophosphate temephos was detected at one site in East Baton Rouge Parish in the spring of 2016, which increased to eightfold resistance by the end of the mosquito season. Activities of esterases were found to be elevated in wild, temephos-resistant mosquitoes, indicating the potential role of these enzymes as a mechanism of resistance. The results of this study provide a baseline of comparison for future measurements of susceptibility in Cx. quinquefasciatus in Louisiana, and may help inform local mosquito control districts as to the effectiveness and sustainability of their insecticide programs.

Gene introgression between related pest populations is an important component in the assessment of how rapidly economically important traits, such as pesticide resistance, can spread within a region. An example of this is provided by the noctuid moth Spodoptera frugiperda (J.E. Smith), or fall armyworm, which is composed of two ‘host strains’ that differ in their host plant preferences. Resistance to a Bacillus thuringiensis toxin has been observed in some populations and there is concern about its spread throughout the Western Hemisphere. If this trait is easily transmitted between strains, it would expand the range of plants affected and make control efforts more difficult. A complicating factor is that the strains are morphologically indistinguishable and can only be identified by a small number of genetic markers. As a result, little is known about the frequency of interstrain hybridization in the wild. This study uses a novel strategy involving comparisons between two single-nucleotide polymorphisms (SNPs) to quantify the frequency of interstrain mating in field populations.The results demonstrate that hybridization between strains is 4- to 5-fold reduced compared to that within strains. In addition, it appears that directional interstrain mating biases observed in laboratory studies are probably not a major factor in determining the distribution of hybrid genotypes in field populations. The differential SNP technique is a significant improvement over current methods for identifying interstrain hybrids and should facilitate our understanding of fall armyworm strain and hybrid distributions in the field and the frequency of genetic exchanges between strains.

Leaffooted plant bugs (LFPBs) (Leptoglossus spp., Guérin-Méneville) (Hemiptera: Coreidae) are large seed-feeding bugs native to the Western Hemisphere. In California, several Leptoglossus spp. feed on almonds, pistachios, and pomegranate and are occasional pests.The objective of this study was to survey the different species of Leptoglossus present in almond, pistachio, and pomegranate orchards in the Central Valley of California. We used two molecular markers, amplified fragment length polymorphisms (AFLPs) and mitochondrial DNA COI, to determine the number of species or strains of each species, and to infer whether individuals of each species move and possibly interbreed with populations from the other host plants. Two species of leaffooted bugs were abundant, Leptoglossus clypealis Heidemann, and Leptoglossus zonatus (Dallas). L. clypealis was collected in almond and pistachio, while L. zonatus was found on all three host plants, but was the dominant species in pomegranate. The AFLP results indicated that L. clypealis consisted of one species, which suggests it moves between almonds and pistachios during the growing season. Mitochondrial DNA COI for L. clypealis found 1–2% divergence between sequences, and a high haplotype diversity of 0.979 with 17 haplotypes. The AFLP results for L. zonatus found two genetically divergent populations which were morphologically similar. The mtDNA COI sequences for L. zonatus were used for haplotype analysis; three haplotypes were found in California, with one haplotype shared with collections from Brazil. The importance of genetic variability and cryptic species for pest management are discussed.

Stink bugs (Hemiptera: Pentatomidae) are an economically important group of insects that attack numerous crops in the central valley of California. Management of these pests using pheromones or biological control can be species specific, and proper identification of insect species is essential for effective management.The objective was to examine genetic variability in four species of stink bugs, which included Chinavia hilaris (Say) (= Acrosternum hilare) (Hemiptera: Pentatomidae), Chlorochroa uhleri (Stål) (Hemiptera: Pentatomidae), Chlorochroa sayi (Stål) (Hemiptera: Pentatomidae), and Thyanta pallidovirens (Stål) (Hemiptera: Pentatomidae) and to determine whether there may be cryptic species present. Stink bugs were collected in pistachios or on adjacent vegetation when abundant in the central valley of California. The mitochondrial DNA cytochrome oxidase I (COI) gene region (i.e., the barcode) was sequenced for each individual. Data were combined with available GenBank accessions for each species and used to construct a phylogenetic tree. Divergence between genera ranged from 11.2 to 15.7%, whereas divergence between the two Chlorochroa spp. was 4.6%. Genetic variation within Chinavia hilaris collections was up to 4.7%, which suggests the presence of a cryptic species. Genetic divergence was highest between individuals of Chinavia hilaris from the west coast and the east coast of the United States. In contrast, genetic variation within individuals of C. uhleri and Ch. sayi was less than 1%. Nine haplotypes were found for Chinavia hilaris, five for C. uhleri, three for Ch. sayi, and five for T. pallidovirens. The relevance of correct species identification and genetic diversity to stink bug management practices was discussed.

The apple maggot fly, Rhagoletis pomonella Walsh (Diptera: Tephritidae), is a serious quarantine pest in the apple-growing regions of central Washington and Oregon. The fly is believed to have been introduced into the Pacific Northwest via the transport of larval-infested apples near Portland, Oregon, within the last 40 yr. However, R. pomonella also attacks native black hawthorn, Crataegus douglasii Lindley (Rosales: Rosaceae), and introduced ornamental hawthorn, Crataegus monogyna Jacquin, in the region. It is, therefore, possible that R. pomonella was not introduced but has always been present on black hawthorn. If true, then the fly may have independently shifted from hawthorn onto apple in the Pacific Northwest within the last 40 yr after apples were introduced. Here, we test the introduction hypothesis through a microsatellite genetic survey of 10 R. pomonella sites in Washington and 5 in the eastern United States, as well as a comparison to patterns of genetic variation between populations of Rhagoletis cingulata Loew and Rhagoletis indifferens Curran, two sister species of cherry-infesting flies known to be native to the eastern and western United States, respectively. We report results based on genetic distance networks, patterns of allelic variation, and estimated times of population divergence that are consistent with the introduction hypothesis for R. pomonella. The results have important implications for R. pomonella management, suggesting that black hawthorn-infesting flies near commercial apple-growing regions of central Washington may harbor sufficient variation to utilize apple as an alternate host, urging careful monitoring, and possible removal of hawthorn trees near orchards.

The silverY moth, Autographa gamma L. (Noctuidae: Plusiinae), is a pest of major economic importance in its native range of Europe, Asia, and North Africa. Although not present in North America, larvae of A. gamma are commonly intercepted in commodity shipments at U.S. ports, and adult surveys are conducted each year in more than 20 states. Because of the similarity of A. gamma to several native North American species that are attracted to the same pheromone lure, morphological identification of adults is difficult and requires dissection. In 2010, a specimen of Autographa californica (Speyer, 1875) (Lepidoptera: Noctuidae) from Pennsylvania was incorrectly identified as A. gamma, signaling the need for an alternative method of rapid identification. Here we detail a real-time PCR assay capable of identifying A. gamma specimens in approximately 45 min using extracted DNA. The assay uses a hydrolysis probe that targets a species-specific segment of the CO1 DNA barcode region, while a control probe targets a conserved region of 18S rDNA. The assay was tested with two independent runs of 452 specimens of Plusiinae representing 23 different species. The assay provided unambiguous data 99.7% of the time and did not result in any false positives; these data were used to develop a rule set for interpreting the real-time PCR results. In addition, the same diagnostic probe was tested in bulk sample simulations using real-time PCR and droplet digital PCR where A. gamma could be detected in concentrations as low as 1:1,000,000 (gamma:californica).These experiments provide baseline data for developing a bulk sample assay.

The superfamily Psylloidea (Hemiptera: Sternorrhyncha) lacks a robust multigene phylogeny. This impedes our understanding of the evolution of this group of insects and, consequently, an accurate identification of individuals, of their plant host associations, and their roles as vectors of economically important plant pathogens.The conserved nuclear gene elongation factor-1 alpha (EF-1α) has been valuable as a higher-level phylogenetic marker in insects and it has also been widely used to investigate the evolution of intron/exon structure. To explore evolutionary relationships among Psylloidea, polymerase chain reaction amplification and nucleotide sequencing of a 250-bp EF-1α gene fragment was applied to psyllids belonging to five different families. Introns were detected in three individuals belonging to two families. The nine genera belonging to the family Aphalaridae all lacked introns, highlighting the possibility of using intron presence/absence as a diagnostic tool at a family level. When paired with cytochrome oxidase I gene sequences, the 250 bp EF-1α sequence appeared to be a very promising higher-level phylogenetic marker for psyllids.

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is considered the main vector of the bacteria associated with huanglongbing, a very serious disease that has threatened the world citrus industry. The absence of efficient control management protocols, including a lack of resistant cultivars, has led to the development of different approaches to study this pathosystem. The production of resistant genotypes relies on D. citri gene expression analyses by RT-qPCR to assess control of the vector population. High-quality, reliable RT-qPCR analyses depend upon proper reference gene selection and validation. However, adequate D. citri reference genes have not yet been identified. In the present study, we evaluated the genes EF 1-α, ACT, GAPDH, RPL7, RPL17, and TUB as candidate reference genes for this insect. Gene expression stability was evaluated using the mathematical algorithms deltaCt, NormFinder, BestKeeper, and geNorm, at five insect developmental stages, grown on two different plant hosts [Citrus sinensis (L.) Osbeck (Sapindales: Rutaceae) and Murraya paniculata (L.) Jack (Sapindales: Rutaceae)]. The final gene ranking was calculated using RefFinder software, and the V-ATPase-A gene was selected for validation. According to our results, two reference genes are recommended when different plant hosts and developmental stages are considered. Considering gene expression studies in D. citri grown on M. paniculata, regardless of the insect developmental stage, GAPDH and RPL7 have the best fit as reference genes in RT-qPCR analyses, whereas GAPDH and EF 1-α are recommended as reference genes in insect studies using C. sinensis.

Generalist insect predators play an essential role in regulating the populations of Bemisia tabaci and other pests in agricultural systems, but may be affected negatively by insecticides applied for pest management. Evaluation of insecticide compatibility with specific predator species can provide a basis for making treatment decisions with the aim of conserving natural enemies. Eleven insecticides representing six modes of action groups were evaluated for toxicity against four predator species and at different developmental stages. Full-concentration series bioassays were conducted on laboratory-reared or insectary-supplied predators using Petri dish and systemic uptake bioassay techniques. Highest toxicities were observed with imidacloprid and clothianidin against first and second instar nymphs of Geocoris punctipes (Say) (Hemiptera: Geocoridae). Later instar nymphs were less susceptible to neonicotinoid treatments based on higher LC₅₀s observed with imidacloprid, thiamethoxam, and dinotefuran against third or fourth instar nymphs.The pyrethroid insecticide bifenthrin was highly toxic against adults of G. punctipes and Orius insidiosus (Say) (Hemiptera: Anthocoridae). Standard concentration/mortality evaluation of nonacute toxicity insecticides, including buprofezin, pyriproxyfen, spirotetramat, and spiromesifen, was inconclusive in terms of generating probit statistics. However, low mortality levels of insects exposed for up to 120 h suggested minimal lethality with the exception of pyriproxyfen that was mildly toxic to Chrysoperla rufilabris (Burmeister) (Neuroptera: Chrysopidae).

Citrus (Citrus spp.) production continues to decline in East Africa, particularly in Kenya andTanzania, the two major producers in the region. This decline is attributed to pests and diseases including infestation by the African citrus triozid, Trioza erytreae (Del Guercio) (Hemiptera:Triozidae). Besides direct feeding damage by adults and immature stages, T. erytreae is the main vector of ‘Candidatus Liberibacter africanus’, the causative agent of Greening disease in Africa, closely related to Huanglongbing. This study aimed to generate a novel barcode reference library for T. erytreae in order to use DNA barcoding as a rapid tool for accurate identification of the pest to aid phytosanitary measures. Triozid samples were collected from citrus orchards in Kenya, Tanzania, and South Africa and from alternative host plants. Sequences generated from populations in the study showed very low variability within acceptable ranges of species. All samples analyzed were linked to T. erytreae of GenBank accession number KU517195. Phylogeny of samples in this study and other Trioza reference species was inferred using the Maximum Likelihood method. The phylogenetic tree was paraphyletic with two distinct branches. The first branch had two clusters: 1) cluster of all populations analyzed with GenBank accession of T. erytreae and 2) cluster of all the other GenBank accession of Trioza species analyzed except T. incrustata Percy, 2016 (KT588307.1), T. eugeniae Froggatt (KY294637.1), and T. grallata Percy, 2016 (KT588308.1) that occupied the second branch as outgroups forming sister clade relationships.These results were further substantiated with genetic distance values and principal component analyses.

Capturing the complementary strengths of observational and experimental research methods usually requires the researcher to gather separate experimental and observational data sets. In some cases, however, commercial agricultural practices produce the spatial and temporal mixing of ‘treatments' independently of other possibly covarying factors that is normally achieved only with formal experimentation. The resulting ‘pseudoexperiments’ can provide strong evidence for causal relationships. Here, we analyze a large observational data set that creates a series of such pseudoexperiments to assess the effect of different commercial varieties of almond, Prunus dulcis (Mill.) on the impact of two key lepidopteran pests, the navel orangeworm Amyelois transitella (Walker) (Lepidoptera: Pyralidae), and the peach twig borer Anarsia lineatella Zeller (Lepidoptera: Gelechiidae). Almonds are universally planted as polycultures of different varieties to obtain efficient cross-pollination. We find substantial differences across almond varieties in the rates of infestation of almond hulls and nutmeats by the two pests. We find no support for the hypothesis that earlier-maturing varieties sustain higher attack; for A. transitella, later-maturing varieties instead had more frequent infestation. On many almond varieties, A. lineatella reaches high infestation levels by feeding almost exclusively on the hulls, rather than nutmeats. Given the importance of these pests in directly destroying almond nuts and in promoting aflatoxin-producing Aspergillus sp. fungal infections of almonds, further work exploring the impact of these pests is warranted. Because many crops requiring cross-pollination are planted as mixtures of different varieties, commercial agricultural production data hold great potential for studying within-crop variation in susceptibility to insect attack.

Understanding how host-crop genotypes affect the life history of insect pests is important for developing and using varietal resistance as a pest control measure. Here we determined how wild and cultivated sugarcane genotypes affect key life-history traits and the demographic performance of the root spittlebug, Mahanarva fimbriolata (Stål; Hemiptera: Cercopidae), a major pest of sugarcane, which produces most of the world's sugar and bioethanol. In the greenhouse, plants of four sugarcane genotypes (two wild and two cultivated) were infested with newlyecloded spittlebug nymphs. A longitudinal life-table experiment was conducted recording developmental time, survival, reproductive output, and longevity of the released spittlebug nymphs. One of the promising sugarcane genotypes was the accession IM76-229 (Saccharum robustum Brandes & Jesw. ex Grassl), which allowed only 20% of nymph survival to adulthood and reduced the spittlebug longevity by 10 d. Such effects on these life-history traits led to a negative intrinsic rate of population growth (r_m = –0.002) of the insects, indicating that the test population would eventually go extinct on this sugarcane genotype. An opposite trend (i.e., population growth) was obtained for the insects raised on Saccharum spp. (SP81-3250 and SP80-1816 cultivars) and Erianthus arundinaceus (Retz) Jeswiet (Kawandang genotype).These results show that S. robustum IM76-229 genotype has resistance traits that drastically reduce spittlebug population growth and indicate that it can be used in breeding programs aiming to develop sugarcane cultivars with resistance to spittlebugs.

Conventional insecticide assays, which measure the effects of insecticide exposure on short-term mortality, overlook important traits, including persistence of toxicity or sub-lethal effects. Therefore, such approaches are especially inadequate for prediction of the overall impact of insecticides on beneficial arthropods. In this study, the side effects of four modern insecticides (chlorantraniliprole, emamectin benzoate, spinosad, and spirotetramat) on Adalia bipunctata (L.) (Coleoptera: Coccinellidae) were evaluated under laboratory conditions by exposition on treated potted plants. In addition to investigation of acute toxicity and persistence of harmful activity in both larvae and adults of A. bipunctata, demographic parameters were evaluated, to provide a comprehensive picture of the nontarget effects of these products. Field doses of the four insecticides caused detrimental effects to A. bipunctata; but in different ways. Overall, spinosad showed the best toxicological profile among the products tested. Emamectin benzoate could be considered a low-risk insecticide, but had high persistence. Chlorantraniliprole exhibited lethal effects on early instar larvae and adults, along with a long-lasting activity, instead spirotetramat showed a low impact on larval and adult mortality and can be considered a short-lived insecticide. However, demographic analysis demonstrated that chlorantraniliprole and spirotetramat caused sub-lethal effects. Our findings highlight that sole assessment of mortality can lead to underestimation of the full impact of pesticides on nontarget insects. Demographic analysis was demonstrated to be a sensitive method for detection of the sub-lethal effects of insecticides on A. bipunctata, and this approach should be considered for evaluation of insecticide selectivity.

The Neotropical brown stink bug Euschistus heros (Fabr.) (Hemiptera: Pentatomidae) is one of the major soybean pests in South America.This species is responsible for high levels of infestation, causes critical damage to seeds, it is associated with leaf retention on plants, and it is extremely difficult to manage. Host plant resistance is a notable technique to assist in reducing the stink bug population in soybean crops. The antibiosis resistance of soybean genotypes to E. heros was evaluated in laboratory. Genotypes L1-1-01, ‘IAC 100', ‘IAC 23', and ‘Coodetec 208’ increased the length of nymphal developmental time. PI 274453 and PI 227687 reduced egg viability and also adult body weight. PI 274454, ‘IAC 19’, PI 227687, and PI 229358 led to low nymphal viability.These results suggest that these genotypes may be useful in soybean breeding programs that focus on the development of genotypes resistant to E. heros.

The isopod, Armadillidium vulgare (Latreille) (Isopoda: Armadillidiidae), and the millipede, Ommatoiulus moreletii (Lucas) (Diplopoda: Iulidae), are increasingly being reported as pests of emerging broadacre crop seedlings in southern Australia. This is thought to be due to the increased adoption of stubble retention practices, leading to increased abundance of these soil-dwelling organisms. Here, we evaluate the propensity of A. vulgare and O. moreletii to damage a range of crop seedlings. Through the combined analysis of a controlled feeding trial and field reports, we show A. vulgare is able to feed on and damage a range of pulses, legumes, cereals, and oilseeds, as emerging seedlings. O. moreletii had a more restricted range of feeding, being limited to lupin, lucerne, and canola in the feeding trial. These results are discussed in the context of developing pest management guidelines for these species.

The aim of this study was to compare the effect of spittlebug Mahanarva fimbriolata Stål (Hemiptera: Cercopidae) on photosynthetic parameters of both a susceptible (SP81-3250) and a resistant (H.Kawandang) sugarcane genotype. In the first assay, the susceptibility level of genotypes to spittlebug was confirmed by comparing damage score and chlorophyll content of the plants. In the second assay, the effect of spittlebug nymphs on photosynthetic characteristics was assessed using the following parameters: Net photosynthetic rate (A), carboxylation efficiency (A/Ci), stomata conductance (g_S), transpiration (E), electron transport rate (ETR), maximum quantum yield of Photosystem 2 (PSII) (F_V/F_M), effective quantum yield (Y(II)), photochemical quenching (Y(NPQ)), and nonphotochemical quenching (Y(NO)). Spittlebug nymphs affected the photosynthetic process of the susceptible genotype SP81-3250 by decreasing the Chl content, ETR, F_V/F_M, andY(II). However, this genotype was able to maintain A probably due to its ability to maintain stomata aperture, increase the carboxylation efficiency of Rubisco, and dissipate excess energy through the xanthophyll cycle, as Y(NPQ) increased under the spittlebug attack. On the other hand, the spittlebug did not affect Chl content and F_V /F_M of the H.Kawandang genotype. Furthermore, H.Kawandang increased A to compensate for the sink demand by the spittlebug by increasing stomatal aperture and carboxylation efficiency and increasing efficiency of the photochemical apparatus in converting light energy into chemical products. We can conclude that the feeding habits of spittlebug nymphs have different impacts on photosynthesis of susceptible and resistant sugarcane genotypes.

Despite decades of research on management tactics for the navel orangeworm, Amyelois transitella (Walker) (Lepidoptera: Pyralidae), on almonds, we still do not have an established means of using in-season pest-density estimates to predict damage to nuts at harvest. As a result, hull-split pesticide applications, although timed carefully to coincide with navel orangeworm oviposition and with crop vulnerability, are not tied to pest densities—thus falling short of our goals under modern pest management. Here we use an ecoinformatics approach, analyzing a pre-existing data set collected in commercial almond production in California, to ask: 1) are navel orangeworm density estimates obtained using different sampling methods in strong agreement with one another? and 2) can we use either single density estimates or combinations of density estimates to explain variation in nutmeat damage at harvest? We find that correlations between density estimates of navel orangeworm made over a single growing season are often weak, and suggest that density estimates taken closer to the time of harvest (catches of adult females between hull split and harvest; infestation of early-split nuts) may be most useful for predicting damage at harvest. Single-density estimates explained ≤39.1% of variation in harvest damage, whereas a combination of predictors explained 51.5% of the total variance in nutmeat damage at harvest. Our results suggest that density estimates taken just prior to harvest may, with refinement, be usable within a predictive framework to guide late-season control decisions.

Delottococcus aberiae De Lotto (Hemiptera: Pseudococcidae) is the latest exotic mealybug species introduced in citrus in the Mediterranean basin. It causes severe distortion and size reduction on developing fruits. Due to its first interaction with citrus, D. aberiae economic thresholds are still unknown for this crop and the current Integrated Pest Management programs have been disrupted.The objectives of this study were to determine the aggregation patterns of D. aberiae in citrus, develop an efficient sampling plan to assess its population density, and calculate its Economic and Economic Environmental Injury Levels (EIL and EEIL, respectively). Twelve and 19 orchards were sampled in 2014 and 2015, respectively. At each orchard, population densities were monitored fortnightly in leaves, twigs, and fruit, and fruit damage was determined at harvest. Our results showed a clumped aggregation of D. aberiae in all organs with no significant differences between generations on fruit. Fruit damage at harvest was strongly correlated with fruit occupation in spring. Based on these results and using chlorpyrifos as the insecticide of reference, the EIL and EEIL were calculated as 7.1 and 12.1% of occupied fruit in spring, respectively. With all this, we recommend sampling 275 fruits using a binomial sampling method or alternatively, 140 fruits with an enumerative method bimonthly between petal fall and July.

Hermetic storage is of interest to farmers and warehouse managers as a method to control insect pests in small storage facilities. To develop improved understanding of effects of hermetic storage on insect pest activity and mortality over time, oxygen levels, acoustic signals, and observations of visual movement were recorded from replicates of 25, 50, and 100 adult Sitophilus oryzae (L.) (Coleoptera: Curculionidae) hermetically sealed in 500- and 1,000-ml glass jars. Recordings were done for 28 d; twice daily for the first 6 d and twice weekly thereafter. Insect sounds were analyzed as short bursts (trains) of impulses with spectra that matched average spectra (profiles) of previously verified insect sound impulses. Oxygen consumption was highest in treatments of 100 insects/500-ml jar and lowest in 25/1000-ml jars. The rates of bursts per insect, number of impulses per burst, and rates of burst impulses per insect decreased as the residual oxygen levels decreased in each treatment. Activity rates <0.02 bursts s^–1, the acoustic detection threshold, typically occurred as oxygen fell below 5%. Mortality was observed at 2% levels. The time to obtain these levels of insect activity and oxygen depletion ranged from 3–14 d depending on initial infestation levels. Acoustic detection made it possible to estimate the duration required for reduction of insect activity to levels resulting in negligible damage to the stored product under hermetic conditions. Such information is of value to farmers and warehouse managers attempting to reduce pest damage in stored crops.

Control of the lesser mealworm (Alphitobius diaperinus Panzer, 1797 [Coleoptera: Tenebrionidae]) is usually conducted with chemical insecticides, which have limited efficacy as well as environmental and health risks. Diatomaceous earth (DE) is safe and effective in controlling many insects including lesser mealworm. However, little is known of DE efficacy against this pest. Thus, an assessment of DE was conducted for the first time in commercial broiler houses in Brazil, with the aim of developing a strategy for the use of DE in the control of this insect. The efficacy of DE was determinate to a minimum concentration, and the application was directed to the most infested sites. The DE applied to the entire poultry house (280 g/m²) was verified to control the insects. DE was also comparable to chemical treatments, with a reduction of 80% in the insect population. DE controlled the lesser mealworm and is recommended for application at a concentration of 280 g/m² when applied to the surface of new poultry litter, before the first lot of birds is housed.Thereafter, DE should be reapplied before the housing of each lot of birds, in the same concentration, only in the housing area and under the feeders and near the walls and pillars. In addition to being a control strategy, DE can be an important tool in the management of lesser mealworm insecticide resistance.

The study of the technical and economic aspects of rearing natural enemies is essential for its effective use as a biological control agent in the field.The aim of this study was to determine the cost of production of the parasitoid, Telenomus remus (Nixon; Hymenoptera: Platygastridae), reared in eggs of its natural host, Spodoptera frugiperda (J. E. Smith; Lepidoptera: Noctuidae), and the alternative host, Corcyra cephalonica (Stainton; Lepidoptera: Pyralidae). The insects were reared in accordance with their respective methodologies of laboratory rearing. The cost of production of this biocontrol agent is US$ 0.0004 when reared with S. frugiperda eggs and US$ 0.0002 with C. cephalonica eggs.

Squash bug, Anasa tristis DeGeer (Hemiptera: Coreidae), is a major pest of squash and pumpkins in the United States. In order to better understand the importance of natural egg parasitism of this species in Virginia, we conducted a 2-yr statewide survey. In total, 1,127 squash bug egg masses (∼20,000 total eggs) were sampled from squash and pumpkins from 43 counties in Virginia from 2014 to 2015. Egg masses were brought back to the lab to record levels of squash bug nymphal emergence or adult parasitoid eclosion and identification. Over 50% of the total squash bug eggs collected statewide were parasitized. Gryon pennsylvanicum Ashmead (Hymenoptera: Scelionidae) was the predominant egg parasitoid accounting for over 98% of all parasitoid adults recovered. The only other species emerging from squash bug eggs was Anastatus reduvii Howard (Hymenoptera: Eupelmidae), which is a generalist parasitoid. G. pennsylvanicum was found in 75% of the counties surveyed with the highest levels of parasitism occurring in the Northern, Southwestern Mountain, and Western Piedmont regions of the state and the lowest levels of parasitism occurring in theTidewater region in the southeastern portion of the state. Based on this 2-yr survey, G. pennsylvanicum was determined to be a major natural enemy of squash bug, significantly reducing the number of nymphs that emerge from deposited eggs. Conservation of this natural enemy should therefore be a priority for integrated pest management programs in cucurbits.