The microsporidia Nosema apis (Zander) and Nosema ceranae (Fries) are common intestinal parasites in honey bee (Apis mellifera L.) colonies. Though globally prevalent, there are mixed reports of Nosema infection costs, with some regions reporting high parasite virulence and colony losses, while others report high Nosema prevalence but few costs. Basic and applied studies are urgently needed to help beekeepers effectively manage Nosema spp., ideally through an integrated pest management approach that allows beekeepers to deploy multiple strategies to control Nosema when Nosema is likely to cause damage to the colonies, rather than using prophylactic treatments. Beekeepers need practical and affordable technologies that facilitate disease diagnosis and science-backed guidelines that recommend when, if at all, to treat infections. In addition, new treatment methods are needed, as there are several problems associated with the chemical use of fumagillin (the only currently extensively studied, but not globally available treatment) to control Nosema parasites. Though selective breeding of Nosema-resistant or tolerant bees may offer a long-term, sustainable solution to Nosema management, other treatments are needed in the interim. Furthermore, the validation of alternative treatment efficacy in field settings is needed along with toxicology assays to ensure that treatments do not have unintended, adverse effects on honey bees or humans. Finally, given variation in Nosema virulence, development of regional management guidelines, rather than universal guidelines, may provide optimal and cost-effective Nosema management, though more research is needed before regional plans can be developed.

Mealybugs (Hemiptera: Pseudococcidae) represent a serious threat for viticulture as vectors of phloem-restricted viruses associated with the grapevine rugose wood and leafroll diseases. Heliococcus bohemicus (Šulc) is known to be involved in the spread of these two viral diseases, being a vector of the Grapevine virus A (GVA) and the Grapevine leafroll-associated virus 1 and 3 (GLRaV-1 and GLRaV-3). This study investigated the acquisition and transmission efficiency of H. bohemicus fed on mixed-infected plants. Nymphs were field-collected onto GVA, GLRaV-1, and GLRaV-3 multiple-infected grapevines in two vineyards in North-Western Italy, and were used in transmission experiments under controlled conditions. Even if most of the collected nymphs were positive to at least one virus, transmission occurred only to a low number of test grapevines. The transmission frequency of GLRaV-3 was the highest, whereas GVA was transmitted to few test plants. The transmission of multiple viruses occurred at low rates, and nymphs that acquired all the three viruses then failed to transmit them together. Statistical analyses showed that the three viruses were independently acquired and transmitted by H. bohemicus and neither synergistic nor antagonistic interactions occurred among them. GVA and GLRaVs transmission efficiencies by H. bohemicus were lower than those reported for other mealybug vectors. This finding is consistent with the slow spread of leafroll and rugose wood diseases observed in Northern Italy, where H. bohemicus is the predominant vector species.

A study on seasonal abundance of Auchenorrhyncha species and their infectivity by Xylella fastidiosa in the Apulia region of Italy was conducted to identify ideal periods for monitoring and adoption of potential control measures against insect vectors. Adult populations of Auchenorrhyncha species were monitored monthly over a 2-yr period from five olive groves. A total of 15 species were captured, identified, and tested for presence of X. fastidiosa by polymerase chain reaction (PCR). For three species, Philaenus spumarius L., Neophilaenus campestris (Fallèn), and Euscelis lineolatus Brullé, positive reactions to X. fastidiosa were obtained, on average, in 16.3, 15.9 and 18.4% of adult insects, respectively. Philaneous spumarius was the dominant species (39.8% of total Auchenorrhyncha captured) with the highest adult abundance in summer months. Adult P. spumarius and N. campestris were first detected between March and May in both years, and all insects tested during these periods (year 1: n = 42, year 2: n = 132) gave negative reactions to X. fastidiosa by PCR. Similarly, first adults of E. lineolatus that appeared from October to November (year 1: n = 20, year 2: n = 15) tested negative for presence of X. fastidiosa. Given the lack of transstadial and transovarial transmission of X. fastidiosa and considering that P. spumarius is univoltine, control measures against nymphal stages of P. spumarius should be investigated as means of population suppression to reduce spread of X. fastidiosa in olive groves.

Several mealybug species are vectors of grapevine leafroll-associated viruses (GLRaV), which cause the economically important grapevine leafroll disease in grape-producing regions worldwide. The mealybug Ferrisia gilli Gullan is a new pest of grapevines in El Dorado County, located in the Sierra Foothill wine-growing region of California. GLRaV species 1, 2, 3, and 4LV have been detected in vineyards with symptomatic vines in the Sierra Foothills. We conducted controlled virus acquisition and transmission experiments using source vine accessions infected with different combinations of GLRaV. We determined that F. gilli acquired GLRaV 1, 2, 3, and 4LV, and transmitted GLRaV-3 and GLRaV-4LV to uninfected recipient vines. Like numerous other mealybug species, in addition to causing direct damage to vines, F. gilli poses a threat to the grape industry as a vector of economically damaging viruses.

Parasitoids can utilize the nutrition of host eggs to culture their own offsprings. Host qualities, including size, age, and species, influence parasitism preference and fitness of the parasitoid offsprings. However, an interesting question here is whether the parasitoids have the ability of recognizing fertilized and nonfertilized host eggs. In this study, we investigated the performance of three indigenous Trichogramma species (Trichogramma japonicum Ashmead, Trichogramma dendrolimi Matsumura, and Trichogramma chilonis Ishii) on fertilized and nonfertilized eggs of their native host, Chilo suppressalis (Walker). The results showed that the three Trichogramma species tested could recognize fertilized and nonfertilized eggs of C. suppressalis. The different fertilization status eggs of C. suppressalis were all accepted by T. japonicum, T. chilonis, and T. dendrolimi with a clear tendency that they preferred to parasitize fertilized eggs whether under no-choice or choice conditions. All Trichogramma species successfully completed development in parasitized eggs of C. suppressalis regardless if the host eggs were fertilized or not. There was similar adult emergence rate, development time, and female progeny between fertilized and nonfertilized eggs for all Trichogramma species with an exception of development for T. chilonis. Trichogramma chilonis had shorter developmental time in fertilized eggs than in nonfertilized ones. Whether in fertilized or nonfertilized eggs, T. dendrolimi had longer development time than other two Trichogramma species. However, T. dendrolimi had the largest percentage of female progeny on fertilized eggs, followed by T. japonicum, and T. chilonis had the least percentage of female progeny. The present study indicates that Trichogramma can recognize fertilized and nonfertilized host eggs and use the nonfertilized eggs to finish offspring development.

Orius strigicollis (Poppius) is an anthocorid bug with high foraging ability on thrips as well as on mites, and the bug has been considered as a potential biological control agent in Taiwan. Life table and predation studies of O. strigicollis fed on Cadra cautella (Walker) and Tetranychus urticae (Koch) eggs were conducted at 25 ± 1°C. Data were analyzed and compared using TWOSEX-MSChart and CONSUME-MSChart software. O. strigicollis fed on eggs of C. cautella, a substitute prey, showed significantly higher survival rate and developmental rate than individuals fed on their natural prey, T. urticae eggs. The fecundity of O. strigicollis fed on C. cautella eggs was, on average, 13.2 times higher than that of those fed on T. urticae eggs, despite of the fact that during the entire nymphal stage, the consumption rate of O. strigicollis on T. urticae eggs was ca. 9 times higher than on almond moth eggs The conversion rate (i.e., number of prey eggs needed to produce one predator egg) for this predatory bug reared on T. urticae eggs and almond moth eggs were 604.6 and 6.0, respectively, indicating that almond moth eggs served as an effective alternative prey for ensuring the predator's reproduction. This is the first study pertaining to the population parameters and predation rates of O. strigicollis using the age-stage two-sex approach to describe differences between O. strigicollis populations reared on natural and alternative preys. This information may be useful in mass rearing programs and field application involving this biological control agent.

Aphidius colemani (Viereck) (Hymenoptera: Aphidiidae) is commercially produced and utilized for biological control of peach aphid Myzus persicae (Sulzer) (Hemiptera: Aphididae) on greenhouse crops in many countries. To provide knowledge for the evaluation of parasitoid–host interactions and development of effective mass rearing programs, we investigated how and why host age or size affected fitness gain in A. colemani. We show that the parasitoid was significantly more likely to encounter larger hosts and that an encounter almost always triggered an attack attempt. However, the attack attempt did not proportionally translate into oviposition because larger aphids had greater ability to defend themselves and the parasitoid spent more time in handling larger aphids. The host age at parasitization had no effect on emergence rates and sex ratio of parasitoid progeny, suggesting that pupae and larvae have similar survival rate in hosts of different ages and/or the parasitoid females do not adjust sex allocation based on host size. When parasitizing mid-aged hosts, the parasitoid gained maximum fitness for their progeny in developmental period, body size, and parasitism. Taking all findings together, we suggest that parasitizing mid-aged green peach aphid nymphs is most profitable for A. colemani.

The invasive Asian longhorned beetle, Anoplophora glabripennis (Motschulsky), is a destructive xylophagous forest pest species originating from Asia. Several endemic North American hymenopteran (Braconidae) species in the mid-Atlantic region were capable of attacking and reproducing on A. glabripennis larvae in laboratory bioassays. Ontsira mellipes Ashmead (Hymenoptera: Braconidae) has been continually reared on A. glabripennis larvae at USDA-ARS BIIRU since 2010, and has been identified as a potential new-association biocontrol agent. Two experiments were conducted to investigate parasitism, paralysis, reproductive biology, larval development, and longevity of adult O. mellipes. In the first experiment, pairs of adult parasitoids were given single A. glabripennis larvae every 2 d (along with honey and water) over their lifetimes, while in the second experiment individual parasitoids were observed daily from egg to adult, and adults were subsequently starved. Adults in the first experiment parasitized ∼21% of beetle larvae presented to them throughout their life, and paralysis of larvae occurred 1–2 d after oviposition. More than half of the individual pairs parasitized A. glabripennis larvae, with each female producing around 26 offspring throughout her life. In the second experiment, median development time of O. mellipes from egg to adult was about 3 wk, with five larval instars. Adult O. mellipes that were provided with host larvae, honey, and water lived 9 d longer than host-deprived and starved adults. These findings indicate that mass-rearing procedures for O. mellipes may be developed using the new association host for development of effective biocontrol programs against A. glabripennis.

It is common for a female trichogrammatid to lay more than one egg per host, a phenomenon known as self-superparasitism, which exposes her offspring to intraspecific, intrinsic competition (IIC) with its own siblings. Information about how often self-superparasitism occurs and how IIC interacts with abiotic factors is rare, especially regarding the Neotropical Trichogramma species. Here we determined the frequency of self-superparasitism in Trichogramma atopovirilia Oatman & Platner (Ta) and T. pretiosum Riley (Tp), and the effects of IIC and temperature on the sex ratio, egg-to-adulthood period, and survivorship of both species’ offspring. Individual females were offered eggs of Spodoptera frugiperda (J.E. Smith) for 30 min. A group of parasitized hosts was then dissected for determination of the self-superparasitism frequency, while another group was incubated at 15, 18, 21, 24, 27, 30, and 33°C. High rates of self-superparasitism were found in both Ta (0.55 ± 0.07) and Tp (0.62 ± 0.06). IIC interacted with temperature decreasing Ta and Tp's survivorship, lengthening the egg-to-adulthood period in Tp and shortening it in Ta, and balancing Ta's sex ratio. Based on survivorship rate, Ta and Tp could not be differentiated if their immatures develop in absence of IIC. However, in its presence, Tp was 3 × more likely to survive than Ta at 33°C, while at 15°C Ta survived 2× better than Tp. These results show that self-superparasitism can be very common in both Ta and Tp, and that its effects on the species’ biological traits and competitiveness strongly depend on the IIC–temperature interaction.

A systems approach has been developed for mitigation of risk associated with Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae), in citrus fruit exported from South Africa, as an alternative to a stand-alone cold treatment. This study was undertaken to assess compliance with inspection standards applicable to various steps within the systems approach and to determine its overall efficacy. Larval infestation of fruit was monitored weekly in fruit from 33 orchards, until the time of harvest, postpicking, and postpacking into export cartons. Significant positive regressions were recorded between infestation of fruit during the full monitoring period in the orchard and the last 4 wk before harvest, between the last 4 wk before harvest and on delivery to the packinghouse, and on delivery to the packinghouse and in the packed carton. There was an improvement in the level of compliance with each of these successive steps in the system, thus verifying that the grading and inspection thresholds were appropriately sensitive and confirmed the effectiveness of the system. The overall risk mitigation efficacy of the systems approach was calculated. The calculation included several known compounding under estimations of efficacy. Nonetheless, the proportion of fruit that could be infested with T. leucotreta after application of the systems approach was between P ≤ 5.328 × 10⁻⁶ and P ≤ 8.380 × 10⁻⁷, 6–38 times less than the proportion associated with the probit 9 (P ≤ 3.2 × 10⁻⁵) standard for a stand-alone cold treatment, being three survivors in 100,000 at the 95% confidence level.

Some of South Africa's export markets require postharvest cold treatment of citrus fruit for phytosanitary risk mitigation for Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae). An alternative to a standalone cold treatment may be a reduced intensity cold treatment as a step in a systems approach. For cold treatment trials, large numbers of larvae are required. Due to recent dramatic improvement of T. leucotreta control in the field, sufficient naturally infested citrus fruit are no longer available. Artificial infestation of fruit is not viable due to rapid decay of the fruit. Consequently, it is necessary to use laboratory-reared T. leucotreta larvae in artificial diet. In trials, field-collected larvae from the Eastern Cape were at least as cold-tolerant as those from other regions. Larvae in Navel oranges showed the median level of susceptibility in a range of citrus types evaluated at 6°C, and their use in trials was considered acceptable due to their greater natural susceptibility to T. leucotreta infestation. We demonstrated that larvae at high density in artificial diet were at least as cold-tolerant as larvae at lower densities. When exposed to 2°C for 18 d or longer, larvae in artificial diet as used in the trials were at least as cold-tolerant as larvae in fruit. Very few surviving larvae from fruit completed development, with no subsequent generation. Consequently, it is considered justifiable to conduct cold-treatment trials with laboratory-reared T. leucotreta larvae in artificial diet without risk of underestimating the effect of cold on feral larvae in citrus fruit.

Some of South Africa's citrus export markets require mandatory postharvest cold treatment of citrus fruit as a phytosanitary risk mitigation treatment for Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae). An alternative to this may be partial cold treatment as one of the final steps in a systems approach to mitigate phytosanitary risk. Consequently, the efficacy of such partial cold treatments was evaluated. It was first determined that a 2°C cold treatment was significantly more effective against fourth and fifth instars (the most cold-tolerant instars) than treatments at 3°C and 4°C for a duration of 18 d. Secondly, it was determined that 2°C for 18 d and 1°C for 16 d were similarly effective, but both treatments were significantly more effective than 1°C for 14 d. Mean mortality of fourth and fifth instars treated with 2°C for 18 d in seven replicates from four trials was 99.94%. Finally, it was determined that the inability of the majority of surviving larvae to develop to adulthood would further increase the efficacy of a 2°C for 18 d treatment to 99.96%. Inclusion of reproductive nonviability of survivors increased mortality to 99.99%.

Insect development occurs within a specific temperature range. Constant temperature studies may produce misleading information on the eco-physiological impacts of temperature on the population dynamics of an insect species, as in most natural environments, temperature usually undergoes daily variation. In China, field surveys showed that the decline in the Agasicles hygrophila (Selman & Vogt) (Coleoptera: Chrysomelidae) population from early August to late September in summer resulted in difficulties in effectively controlling the population of Alternanthera philoxeroides (Mart.) Griseb (Amaranthaceae). Previous studies have largely ignored more natural, fluctuating conditions. In our study, we first investigated the impacts of different temperature conditions (25°C constant temperature for 20 h with a 4-h period of a high temperature of either 30°C, 33°C, 36°C, or 39°C) on adult reproduction and longevity, egg development time, egg hatch rate, female ovarian development, and oogenesis of A. hygrophila. Our results indicated that high temperatures of 30°C and 33°C did not affect the female ovarian development and oogenesis of A. hygrophila. Contrarily, high temperatures of 36°C and 39°C negatively affected the population development of A. hygrophila. At 36°C and 39°C, the egg hatch rates were very low, and the egg development times significantly lengthened. The frequency of abnormal ovaries significantly rose at 39°C. We concluded that the decline in the A. hygrophila population during August and September may be related to the extreme high temperatures that frequently occur in summer. These results help provide a better understanding of A. hygrophila population dynamics under natural conditions.

The green peach aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae), is an agricultural pest that seriously infests many crops worldwide. This study used electrical penetration graphs (EPGs) and life table parameters to estimate the sublethal effects of cyantraniliprole and imidacloprid on the feeding behavior and hormesis of M. persicae. The sublethal concentrations (LC₃₀) of cyantraniliprole and imidacloprid against adult M. persicae were 4.933 and 0.541 mg L⁻¹, respectively. The feeding data obtained from EPG analysis indicated that the count probes and number of short probes (<3 min) were significantly increased when aphids were exposed to LC₃₀ of imidacloprid-treated plants. In addition, the phloem-feeding behavior of M. persicae was significantly impaired on fed tobacco plants treated with cyantraniliprole and imidacloprid at LC₃₀. Analysis of life table parameters indicated that the growth and reproduction of F₁ generation aphids were significantly affected when initial adults were exposed to LC₃₀ of cyantraniliprole and imidacloprid. The nymphal period, female longevity, total preoviposition period, and mean generation time were significantly prolonged when initial adults were exposed to LC₃₀ of imidacloprid. By comparison, these parameters were prolonged but not significantly in the cyantraniliprole treatment. The fecundity and gross reproductive rate were significantly increased in the treated groups. Similarly, the net reproductive rate was greater in the treated group than the control group. Our results indicate that treatment with LC₃₀ of imidacloprid and cyantraniliprole would lead to a hormetic response of M. persicae, with higher likelihood of occurrence when initial adults were exposed to LC₃₀ of cyantraniliprole.

To understand the role that temperature and humidity play in the population dynamics of the beet webworm, Loxostege sticticalis L. (Lepidoptera: Crambidae), egg hatch, survival of first–fifth instars, survival of the full larval stage, survival curves, and larval development rates were investigated at combinations of four temperatures (18, 22, 26, and 30°C) and five relative humidities (RH; 20%, 40%, 60%, 80%, and 100%). We found that greatest egg hatch rate, survival rates of the first and second instars, and survival rate of the complete larval stage occurred at 22°C and 60–80% RH; the lowest values for these parameters were observed at 30°C and 20% RH. Survival of first instars was significantly affected by the interaction of temperature and relative humidity. However, survival of third and fourth instars was neither affected by temperature nor relative humidity, and that of fifth instars was significantly affected only by relative humidity level. The survival curve for larvae was well described by a type III Weibull distribution. Duration of larval stage decreased as temperature increased, but was not affected by relative humidity. We therefore conclude that eggs and early instars are the most critical stages for survival to the pupal stage, and 22–26°C and 60–80% RH are the optimum conditions for their survival and development. These findings confirm that temperature and relative humidity are the critical environmental factors affecting the population growth of L. sticticalis, with temperature being more important.

It is known that the release of volatile chemicals by many plants can attract the natural enemies of herbivorous insects. Such indirect interactions are likely when plants produce nectar from their extrafloral nectaries, and particularly when the production of extrafloral nectar (EFN) is induced by herbivory. In the present study, we conducted experiments to test whether foliar herbivory inflicted by Spodoptera frugiperda Smith (Noctuidae) increases nectar production by extrafloral nectaries on one of its host plants, Ricinus communis L. (Euphorbiaceae). Due to the current economic importance of R. communis, we also investigated whether the following seed traits—water content, dry mass, and essential oil production—are negatively affected by herbivory. Finally, we tested whether or not nectar production and seed traits are influenced by plant fertilization (plant quality). We found that nectar production was increased after herbivory, but it was not affected by the type of fertilization. Seed dry mass was higher in plants that were subjected to full fertilization, without herbivory; plants maintained in low fertilization conditions, however, had higher seed mass when subjected to herbivory. The same inverted pattern was observed for oil production. Therefore, our results suggest that EFN production in R. communis may act as an indirect defense strategy against herbivores, and that there is a trade-off between reproduction and plant growth when low-fertilized plants are subjected to herbivory.

While wild silkworms have served humans for several thousand years, little attention on cold hardiness has been paid to these economically important species. In the present study, supercooling capacity and low temperature tolerance of Chinese oak silkworm, Antheraea pernyi (Guérin-Méneville) (Lepidoptera: Saturniidae), an economic insect reared both for silk production as well as human food, were examined under laboratory conditions. The supercooling points (SCPs) of pupae dropped significantly from a mean of −15.6°C in prediapause to −20.1°C in diapause, and then increased to −17.5°C during postdiapause development. Sex and voltinism influenced body mass but had no significant effect on the SCP. Our data demonstrated that cold tolerance of A. pernyi is tightly linked to life stage. Exposure of eggs to −5°C for up to 8 h had no effect on the hatching rate, whereas silkworm larvae failed to break through the chorion and hatch following a 4–8-h exposure to −10°C. Mean SCPs of intact eggs and naked larvae one day before hatching were similar, −23.3°C and −22.3°C, respectively, indicating that chorion does not significantly affect SCP. Comparison of lower lethal temperature (LLT₅₀) and SCP means suggested that both pupae and eggs of A. pernyi are chill intolerant. These data will improve our understanding of low temperature tolerance in this commercially important species.

Ant species like Pheidole megacephala (F.), Solenopsis invicta (Buren), and the Argentine ant, Linepithema humile (Mayr), have repeatedly been reported to be strongly associated with honeydew-producing arthropods like aphids, scale insects, and mealybugs, effectively protecting them from biological control agents like parasitoids. Here we report the results of a successful trial using pheromone dispensers to suppress Argentine ant activity over large sections in a commercial vineyard over a period of two months and preventing ant access into and foraging within the vine canopy. We found Argentine ant activity to be significantly reduced in pheromone-treated plots for the duration of the trial period compared with control plots. Our results showed a significant reduction in the numbers of Argentine ant workers recruited to randomly placed food resources within treated plots compared with untreated plots. Furthermore, spatial distribution of Argentine ants alongside transects in untreated plots remained relatively continuous, while increasing sharply beyond the borders of treated plots. Lastly, we measured the body fat content of workers and found a significant reduction in fat among workers from treated plots compared with untreated plots, suggesting an adverse effects on nest fitness. Additionally, we provide an initial assessment of the feasibility of the presented approach. Our results showed that it is possible to control Argentine ant, preventing them access to and foraging within the vine canopy, thereby reducing Argentine ants’ access to honeydew.

The white-backed planthopper, Sogatella furcifera (Horváth), is a destructive migratory pest in east and southeast Asia. Huge populations stemming from annual migrations by this insect have caused a series of devastating losses to rice production. There have been numerous early immigrations in five of the past 10 yr but few early immigrations in the others. The annual fluctuation in early immigration is evident, but the mechanism behind these annual fluctuations is unclear. This research aimed to determine the underlying causes for the annual fluctuations in early immigration. We used trajectory analysis to explore the source areas and investigated the meteorological conditions to determine the reason for the annual fluctuations. The results showed that 1) the source areas of S. furcifera are mainly located west of Guangdong and east of Guangxi; 2) the annual fluctuations of the immigrant population size is significantly correlated with the frequency of prevailing winds; and 3) early immigration is influenced by both winter and spring temperatures in the south central Indochina peninsula. These results indicated that an allopatric prediction and sustainable management of rice planthoppers would be difficult to implement within one country. International cooperation and information exchange about this pest between China and other countries in Southeast Asia should be implemented.

The almond wasp, Eurytoma amygdali Enderlein (Hymenoptera: Eurytomidae), a key pest of almond, is a univoltine pest diapausing as last instar larvae inside the damaged fruits for almost nine months in a year. In this study, changes in the amount of total simple sugars, lipid, protein, glycogen, trehalose, glucose, supercooling points (SCPs), and cold hardiness of the diapausing larvae were measured from October to March for first year diapause-destined and in August and September for second year diapause-destined larvae. Changes in glycogen content were reversely proportional to changes in total simple sugars and low molecular weight carbohydrates. These changes reflect the interconversion of glycogen to sugar alcohol in order to increase the insect cold tolerance. We found that cold hardiness and diapause of the last instar larvae of the almond wasp have evolved separately. Cold hardiness was highly associated with physiological changes (accumulation of cryoprotectants), but no physiological changes occurred in early diapause of first year diapause-destined and second year diapause-destined larvae. The almond wasp larvae were found to be a freeze-avoidant insect, as no larva survived after SCP determination and crystallization of its body fluids.

The daily susceptibility rhythm of the cotton aphid, Aphis gossypii Glover, to diazinon and the corresponding changes in the activity of three xenobiotic detoxifying enzymes—cytochrome P450 monooxygenases (P450), glutathione S-transferases (GSTs), and esterases—were investigated. Bioassays were conducted to estimate the median lethal doses (LD₅₀) of diazinon at six different zeitgeber times (ZT0, 4, 8, 12, 16, and 20) under constant light (LL) and lighting conditions of 16 h of light and 8 h of darkness (LD). The results showed that the highest susceptibility occurred at the onset of night (ZT16) and 4 h before this time point (ZT12) under the LD condition. The endogenous rhythmicity of susceptibility was ensured, as the highest susceptibility occurred at the same time under the LL condition. The circadian changes in susceptibility to diazinon were almost coincident with changes in esterase and GSTs activity, but not in P450 activity. We also found rhythmic changes in energy components of whole-body aphids, with similar patterns of circadian changes of proteins, lipids, and soluble carbohydrates, but not glycogen, under LL and LD conditions. These photoperiod conditions (LD and LL) showed different fluctuation in trends of energy resources and of course, different quantities. Our study represents the first report of circadian control of insecticide susceptibility in aphids and provides insights into more efficient control of these pests by unveiling the times of day during which aphids are more susceptible to insecticides with attention to endogenous physiological phenomena.

Linepithema micans (Forel) (Hymenoptera: Formicidae) is the main ant species responsible for dispersal of Eurhizococcus brasiliensis (Wille) (Hemiptera: Margarodidae), a root scale that damages grapevines in southern Brazil. The effects of different formulations of toxic baits based on boric acid and hydramethylnon to control L. micans and E. brasiliensis were evaluated. Toxic baits with boric acid (1.0%) mixed in different concentrations of inverted sugar (20%, 30%, and 40%), and hydramethylnon, mixed with sardines (paste), cassava flour and peanut, brown sugar (sucrose), or sardine oil-based gel, were evaluated in a greenhouse and in the field. In the greenhouse experiment, the number of foraging ants was significantly reduced in the pots where the hydramethylnon in sardine paste (Solid S), sardine oil-brown sugar-based gel (GEL SAM), and peanut oil-brown-sugar gel (GEL AM) formulations were applied. The GEL SAM toxic bait effectively reduced the infestation of L. micans, and could be used for indirect control of E. brasiliensis on young grapevines.

The corn earworm, Helicoverpa zea (Boddie), is a polyphagous pest found throughout the United States, where it attacks many field and vegetable crops. Although H. zea has long been a traditional pest of sweet corn, its importance to this crop has increased dramatically over the past two decades. In this review, we summarize information critical for current and future management of H. zea in sweet corn production in the United States. First, we discuss the pest status of H. zea and its life history, including migration, infestation and larval development, diapause, overwintering, and abiotic factors that affect its biology. Next we describe monitoring methods, crop protection decision-making processes, chemical control options, and the use of genetic technologies for control of H. zea. Alternative H. zea management options including biological control, cultural controls, host plant resistance, and pheromone disruption are also reviewed. The role of climate change and its effects on H. zea and its ecology are discussed, as well as the recent invasion of its relative, Helicoverpa armigera (Hübner), which is a major pest of corn in other parts of the world. To conclude, we suggest future research opportunities for H. zea and H. armigera management in sweet corn.

Previous studies have demonstrated that the agricultural antibiotic jinggangmycin (JGM) stimulates reproduction in the brown planthopper Nilaparvata lugens Stål and that the stimulation of brown planthopper reproduction induced by JGM is regulated by the fatty acid synthase (FAS) and acetyl-CoA carboxylase (ACC) genes. However, a key issue in the stimulation of reproduction induced by pesticides involves the growth and development of oocytes. Therefore, the present study investigated oocyte changes via transmission electron microscopy (TEM) and changes in hormone levels (juvenile hormones (JH) and 20-hydroxyecdysone (20 E)) in JGM-treated females. TEM observations showed that the size of the lipid droplets in the oocytes of JGM-treated females, compared with those in the oocytes of the control females, significantly reduced by 32.6 and 29.8% at 1 and 2 d after emergence (1 and 2 DAE), respectively. In addition, the JH levels of JGM-treated females at 1 and 2 DAE were increased by 49.7 and 45.7%, respectively, whereas 20 E levels decreased by 36.0 and 30.0%, respectively. We conclude that JGM treatments lead to substantial changes in lipid metabolism, which are directly and indirectly related to stimulation of reproduction of brown planthopper together with our previous findings.

Methyl anthranilate was identified as the active compound in extracts of maize (Zea mays L.) roots that were shown to be repellent to neonate western corn rootworm (Diabrotica virgifera virgifera LeConte) larvae. A bioassay-driven approach was used to isolate the active material from diethyl ether extracts of roots from germinating maize seeds. Separation of the extract on a Florisil column yielded an active fraction of 90:10 hexane:diethyl ether. Analysis with gas chromatography–mass spectrometry identified two compounds in the active fraction: indole (2,3-benzopyrrole) and methyl anthranilate (methyl 2-aminobenzoate). When tested in behavioral bioassays, methyl anthranilate elicited a significant (P < 0.05) repellent response at doses of 1, 10, and 100 µg. In subsequent single-choice bioassays, 1, 10, and 100 µg of methyl anthranilate prevented larvae from approaching 10 mmol/mol concentrations of carbon dioxide, which is normally highly attractive to the larvae. Indole, the other compound identified from the active fraction, did not elicit a behavioral response by the larvae. Methyl anthranilate has potential for development as a management tool for western corn rootworm larvae and may be best suited for use in a push–pull control strategy.

Decreased pest pressure is sometimes associated with more diverse agroecosystems, including the addition of a rye cover crop (Secale cereale L.). However, not all pests respond similarly to greater vegetational diversity. Polyphagous pests, such as true armyworm (Mythimna unipuncta Haworth), black cutworm (Agrotis ipsilon Hufnagel), and common stalk borer (Papaipema nebris Guenee), whose host range includes rye have the potential to cause injury to crops following a rye cover crop. The objectives of this study were to compare the abundance of early-season insect pests and injury to corn (Zea mays L.) from fields with and without a rye cover crop on commercial farms. Fields were sampled weekly to quantify adult and larval pests and feeding injury to corn plants from mid-April until corn reached V8 stage, during 2014 and 2015. Measurements within fields were collected along transects that extended perpendicularly from field edges into the interior of cornfields. Adult true armyworm and adult black cutworm were captured around all cornfields, but most lepidopteran larvae captured within cornfields were true armyworm and common stalk borer. Cornfields with a rye cover crop had significantly greater abundance of true armyworm and greater proportion of injured corn. Both true armyworm abundance and feeding injury were significantly greater in the interior of cornfields with rye. Common stalk borer abundance did not differ between cornfields with or without rye cover. Farmers planting corn following a rye cover crop should be aware of the potential for increased presence of true armyworm and for greater injury to corn.

The concept of insecticide termination timing is generally accepted among cotton (Gossypium hirsutum) researchers; however, exact timings are often disputed. Specifically, there is uncertainty regarding the last economic insecticide application to control fruit-feeding pests including tarnished plant bug (Lygus lineolaris (Palisot de Beauvois)), boll weevil (Anthonomus grandis), bollworm (Helicoverpa zea), tobacco budworm (Heliothis virescens), and cotton fleahopper (Pseudatomoscelis seriatus). A systematic review of prior studies was conducted within a meta-analytic framework. Nine publicly available articles were amalgamated to develop an optimal timing principle. These prior studies reported 53 independent multiple means comparison field experiments for a total of 247 trial observations. Stochastic plateau theory integrated with econometric meta-analysis methodology was applied to the meta-database to determine the shape of the functional form of both the agronomic optimal insecticide termination timing and corresponding yield potential. Results indicated that current university insecticide termination timing recommendations are later than overall estimated timing suggested. The estimated 159 heat units (HU) after the fifth position above white flower (NAWF5) was found to be statistically different than the 194 HU termination used as the status quo recommended termination timing. Insecticides applied after 159 HU may have been applied in excess, resulting in unnecessary economic and environmental costs. Empirical results also suggested that extending the insecticide termination time by one unit resulted in a cotton lint yield increase of 0.27 kilograms per hectare up to the timing where the plateau began. Based on economic analyses, profit-maximizing producers may cease application as soon as 124 HU after NAWF5. These results provided insights useful to improve production systems by applying inputs only when benefits were expected to be in excess of the respective costs.

The crucifer flea beetle, Phyllotreta cruciferae (Goeze), is a key pest of canola (Brassica napus L.) in the northern Great Plains of North America. The efficacies of entomopathogenic nematodes (Steinernema spp. and Heterorhabditis spp.), a sprayable polymer gel, and a combination of both were assessed on canola for flea beetle management. Plots were treated soon after colonization by adult flea beetles, when canola was in the cotyledon to one-leaf stage. Ten plants along a 3.6-m section of row were selected and rated at pre-treatment and 7 and 14 d post treatment using the damage-rating scheme advanced by the European Plant Protection Organization, where 1 = 0%, 2 = 2%, 3 = 5%, 4 = 10%, and 5 = 25% leaf area injury. Under moderate flea beetle feeding pressure (1–3.3% leaf area damaged), seeds treated with Gaucho 600 (Bayer CropScience LP Raleigh, NC) (imidacloprid) produced the highest yield (843.2 kg/ha). Meanwhile, Barricade (Barricade International, Inc. Hobe Sound, FL) (polymer gel; 1%) + Scanmask (BioLogic Company Inc, Willow Hill, PA) (Steinernema feltiae) resulted in the highest yields: 1020.8 kg/ha under high (2.0–5.3% leaf area damaged), and 670.2 kg/ha at extremely high (4.3–8.6 % leaf area damaged) feeding pressure. Our results suggest that Barricade (1%) + Scanmask (S. feltiae) can serve as an alternative to the conventional chemical seed treatment. Moreover, Scanmask (S. feltiae) can be used to complement the effects of seed treatment after its protection has run out.

The soybean looper, Chrysodeixis includens (Walker) (Lepidoptera: Noctuidae), is a major defoliating pest of soybeans, Glycine max (L.) Merrill, in Louisiana. However, other alternate host crops in the agroecosystem have the potential to impact C. includens populations. Life table statistics of C. includens on four host plants were evaluated. C. includens larvae were fed leaves of three cotton Gossypium hirsutum L. cultivars ‘DP 143 B2RF,’ ‘DP 174 RF,’ and ‘PHY 485 WRF’; cowpea Vigna unguiculata (L.) Walpers ‘California Blackeye’; three soybean cultivars ‘Lyon,’ ‘PI 227687,’ and ‘RC 4955’; and sweetpotato Ipomoea batatas (L.) Lamarck ‘Evangeline.’ All C. includens larvae reared on cotton cultivars DP 143 B2RF and PHY 485 WRF experienced 100% mortality during the first instar. Total developmental period of preadult C. includens was significantly shorter on cotton DP 174 RF and cowpea California Blackeye but longer on sweetpotato Evangeline. Sweetpotato Evangeline had the highest amount of leaf tissue consumed and soybean Lyon had the least. Pupal weight was highest when insects fed on cotton DP 174 RF and lowest on soybean PI 227687. Life table statistics showed that the highest intrinsic rate of increase and net reproductive rate were attained when insects were reared on cotton DP 174 RF and cowpea California Blackeye whilst the lowest were recorded on soybean PI 227687. This study provides valuable information on the role of alternative host crops on the partial life history of C. includens in Louisiana agroecosystems.

The southern pine beetle Dendroctonus frontalis Zimmermann (Coleoptera: Curculionidae: Scolytinae) is attracted to an aggregation pheromone that includes the multifunctional pheromone component endo-brevicomin. The effect of endo-brevicomin on attractive lures varies from strong enhancement to reduction of beetle attraction depending upon release rate, lure component spacing, and proximity of beetle infestations. Anecdotal observations have further suggested that the effects of endo-brevicomin vary during the year. We investigated this possibility under nonoutbreak conditions in southwestern Mississippi where for two-and-a-half years we monitored traps baited with frontalin and the host odor alpha-pinene either (a) alone, or with an endo-brevicomin release device either (b) located directly on the trap, or (c) displaced 6 m away. The endo-brevicomin devices in our tests increased D. frontalis catches during all times of year, and 6 m displacement of the endo-brevicomin release device from the trap did not significantly alter responses except during the spring flight peak when displacement increased catches. Our data suggest that flying D. frontalis have a stronger tendency to avoid the immediate proximity of a release point of endo-brevicomin during their springtime dispersal flight when catches are greatest. Catches of Thanasimus dubius (F.) (Coleoptera: Cleridae), a major predator of D. frontalis, were not altered by endo-brevicomin, and ratios of D. frontalis to T. dubius changed over the course of the year. We discuss the possible effects of intra-annual variation in D. frontalis response to endo-brevicomin both on beetle attack behavior and use of endo-brevicomin as a lure adjuvant in D. frontalis population monitoring.

Loblolly pine (Pinus taeda L.) is the most important commercial tree species in the southeastern United States. Since the 1950s, there have been reports of loblolly pines showing reduced growth and increased mortality, particularly in central Alabama and western Georgia, United States; the phenomenon is termed as southern pine decline (SPD). Recently, the role of rhizophagous (root-feeding) insects in loblolly pine health within the context of SPD has come under greater scrutiny. We investigated the impacts of subcortical insects, particularly rhizophagous weevils (Coleoptera: Curculionidae), on loblolly pine health in northeastern Georgia. We created plots—representing a gradient of increased relative tree stress—from ungirdled trees, ungirdled trees baited with ethanol and turpentine (ungirdled-baited), and girdled trees. In total, 10,795 subcortical insects from four families (Buprestidae, Cerambycidae, Curculionidae, and Siricidae) and >82 species were trapped in two years. Almost half of the insects trapped (46% of individuals and 11% of species) were nonnative to North America. Insect captures in plots with girdled trees were 61 and 187% greater than those with ungirdled-baited and ungirdled trees, respectively. Tree treatment impacted captures of native, but not nonnative insects. Relative feeding area by the rhizophagous weevils Hylobius pales (Herbst) and Pachylobius picivorus (Germar) on pine twigs placed in pitfall traps was 1, 17, and 82% in plots with ungirdled, ungirdled-baited, and girdled trees, respectively. Hence, there was a strong association of native subcortical insects, especially rhizophagous weevils, with relatively highly stressed trees, confirming that they are secondary instead of primary pine colonizers.

The cerambycid beetles comprise a diverse family that includes many economically important pests of living and dead trees. Pheromone lures have been developed for cerambycids in many parts of the world, but to date, have not been tested in Australia. In this study, we tested the efficacy of several pheromones, identified from North American and European species, as attractants for cerambycids at three sites in southeast Queensland, Australia. Over two field seasons, we trapped 863 individuals from 47 cerambycid species. In the first season, racemic 3-hydroxyhexan-2-one was the most attractive compound among the eight pheromones tested. Subsequently, we aimed to optimize trapping success by combining this compound with other components. However, neither the addition of other pheromone components nor host plant volatiles improved the efficacy of 3-hydroxyhexan-2-one alone. We also tested a generic pheromone blend developed for North American cerambycids, and found that only the combination of this blend with host plant volatiles improved trapping success. The Australian cerambycid fauna is not well known, and effective lures for use in trapping beetles would greatly assist in the study of this important group. Effective semiochemical lures would also have implications for biosecurity through improved monitoring for invasive species.

We assessed how height of panel traps above the forest floor, and the type of trap bait used, influenced the abundance and diversity of cerambycid beetles caught in forested areas of east-central Illinois. Panel traps were suspended from branches of hardwood trees at three heights above the ground: understory (∼1.5 m), lower canopy (∼6 m), and midcanopy (∼12 m). Traps were baited with either a multispecies blend of synthesized cerambycid pheromones or a fermenting bait mixture. Traps captured a total of 848 beetles of 50 species in the cerambycid subfamilies Cerambycinae, Lamiinae, Lepturinae, and Parandrinae, and one species in the closely related family Disteniidae. The species caught in highest numbers was the cerambycine Anelaphus pumilus (Newman), represented by 349 specimens. The 17 most abundant species (mean ± 1 SD: 45 ± 80 specimens per species) included 12 cerambycine and five lamiine species. Of these most abundant species, 13 (77%) were attracted to traps baited with the pheromone blend. Only the cerambycine Eburia quadrigeminata (Say) was attracted by the fermenting bait. Three species were captured primarily in understory traps, and another five species primarily in midcanopy traps. Variation among cerambycid species in their vertical distribution in forests accounted for similar overall abundances and species richness across trap height treatments. These findings suggest that trapping surveys of native communities of cerambycids, and quarantine surveillance for newly introduced exotic species, would be optimized by including a variety of trap baits and distributing traps across vertical strata of forests.

Development of spider mite management technology other than chemical control is desired because of the serious development of acaricide resistance worldwide. Recent studies have evidenced the lethal effects of ultraviolet-B (UVB) radiation on spider mites. To develop the technology on how to use UVB irradiation for spider mite control, we tested whether UVB lamp–light reflection sheet (LRS) combinations suppressed the population size of Tetranychus urticae Koch on strawberry in a greenhouse from December to May (2012–2013, 2013–2014) in Japan. We designed four combinations of UVB lamps and LRSs: 1) neither UVB lamps nor LRSs (UV−LRS−); 2) a UVB lamp without an LRS (UV+LRS−; 2012–2013 only); 3) a UVB lamp and a mulch-type LRS (UV+LRSm); and 4) a UVB lamp and a wing-type LRS (UV+LRSw). The number of adult females peaked at 438.0 and 222.0 per plant in UV−LRS− of 2012–2013 and 2013–2014, respectively, and peaked at 191.6 females in UV+LRS− of 2012–2013. In contrast, the peak abundance was 20.9–98.0 females in UV+LRSm, and fewer than 15 females were noted in UV+LRSw over either experimental period. UVB irradiance on lower leaf surfaces was higher in UV+LRSm and UV+LRSw than UV−LRS− and UV+LRS−, and the mite densities were significantly correlated with UVB irradiance on lower leaf surfaces. Consequently, we conclude that the combinations of UVB lamp–LRS have an excellent capacity to control T. urticae on greenhouse strawberry, and that the LRS was an essential component in this technological approach.

Organic zucchini squash is a high-value vegetable crop in Florida and potential exists to expand its production throughout the state. A lack of knowledge on the effectiveness of organic products and their integration with natural enemies is an important constraint to the regulation of pest populations in organic squash production in Florida. The objectives of this study were to evaluate the effect of insecticides labeled for organic production that can be used for management of Bemisia tabaci (Gennadius) biotype B, on organically grown squash; and to determine the effects of the most efficient insecticides on a key natural enemy, Delphastus catalinae (Horn). Experiments were conducted in the greenhouse in exclusion cages. The first experiment compared the effects of four bioinsecticides on whitefly densities. Insecticides include 1) AzaSol (azadirachtin), 2) PyGanic EC 1.4 (pyrethrin), 3) M-Pede (insecticidal soap), and 4) Entrust (spinosad). The second experiment investigated the effects of bioinsecticides on D. catalinae. Treatment effectiveness was evaluated 1, 3, and 5 d posttreatment. PyGanic and M-Pede were highly effective in controlling whitefly populations on organic squash, while moderate control was provided by AzaSol and there was no control provided by Entrust. PyGanic and M-Pede treatments reduced D. catalinae populations when adults were released 1 d post pesticide application. However, when adults were released 5 d post application, there was no reduction. The importance of using bioinsecticides in combination with natural enemies to regulate pest populations in organic cropping systems is discussed.

Onion thrips, Thrips tabaci Lindeman (Thysanoptera: Thripidae), is a highly destructive pest of onion, Allium cepa L., and its management relies on multiple applications of foliar insecticides. Development of insecticide resistance is common in T. tabaci populations, and new strategies are needed to relax existing levels of insecticide use, but still provide protection against T. tabaci without compromising marketable onion yield. An action threshold-based insecticide program combined with or without a thrips-resistant onion cultivar was investigated as an improved approach for managing T. tabaci infestations in commercial onion fields. Regardless of cultivar type, the average number of insecticide applications needed to manage T. tabaci infestations in the action-threshold based program was 4.3, while the average number of sprays in the standard weekly program was 7.2 (a 40% reduction). The mean percent reduction in numbers of applications following the action threshold treatment in the thrips-resistant onion cultivar, ‘Advantage’, was 46.7% (range 40–50%) compared with the standard program, whereas the percentage reduction in applications in action threshold treatments in the thrips-susceptible onion cultivar, ‘Santana’, was 34.3% (range 13–50%) compared with the standard program, suggesting a benefit of the thrips-resistant cultivar. Marketable bulb yields for both ‘Advantage’ and ‘Santana’ in the action threshold-based program were nearly identical to those in the standard program, indicating that commercially acceptable bulb yields will be generated with fewer insecticide sprays following an action threshold-based program, saving money, time and benefiting the environment.

Spotted-wing drosophila, Drosophila suzukii Matsumura (Diptera: Drosophilidae), is a polyphagous, invasive pest of small fruits. Current management relies heavily on chemical insecticides, and an effective oviposition deterrent could contribute to alternative management approaches that reduce the need for these chemical insecticides. A novel deployment method for repelling Drosophila suzukii, thereby reducing D. suzukii oviposition in fall-bearing red raspberry, was evaluated in the field. Infestations occurring within 4 d after deployment were significantly lower in 2-m-long plots (Rubus idaeus ‘Caroline’) treated with the repellent (20% 1-octen-3-ol in specialized pheromone and lure application technology [SPLAT]) compared to control plots (blank SPLAT). Repellent-treated plots had roughly 28.8 and 49.5% fewer offspring reared per gram of fruit than control plots in two experiments, respectively. Nontarget effects were also evaluated in 2-m plot experiments as well as 5- by 5-m plot experiments. There were no differences in the number of parasitic hymenoptera trapped on yellow sticky cards hung in repellent compared to control plots. While there were no differences in the number of visits to raspberry flowers observed by honey bees in repellent versus control plots, the number of visits by bumble bees was greater in repellent plots compared to control plots. Challenges regarding evaporation rates and potential uses for repellents in an integrated pest management program for the control of D. suzukii are discussed.

Mexican bean beetle, Epilachna varivestis Mulsant, is a serious pest of snap beans, Phaseolus vulgaris L., in the eastern United States. These beetles are intolerant to direct sunlight, explaining why individuals are typically found on the undersides of leaves and in the lower portion of the plant canopy. We hypothesized that snap beans grown on reflective, agricultural polyethylene (plastic mulch) would have fewer Mexican bean beetles and less injury than those grown on black plastic or bare soil. In 2014 and 2015, beans were seeded into beds of metallized, white, and black plastic, and bare soil, in field plots near Blacksburg, VA. Mexican bean beetle density, feeding injury, predatory arthropods, and snap bean yield were sampled. Reflected light intensity, temperature, and humidity were monitored using data loggers. Pyranometer readings showed that reflected light intensity was highest over metallized plastic and second highest over white plastic; black plastic and bare soil were similarly low. Temperature and humidity were unaffected by treatments. Significant reductions in Mexican bean beetle densities and feeding injury were observed in both metallized and white plastic plots compared to black plastic and bare soil, with metallized plastic having the fewest Mexican bean beetle life stages and injury. Predatory arthropod densities were not reduced by reflective plastic. Metallized plots produced the highest yields, followed by white. The results of this study suggest that growing snap beans on reflective plastic mulch can suppress the incidence and damage of Mexican bean beetle, and increase yield in snap beans.

The banana-spotting bug, Amblypelta lutescens lutescens Distant (Hemiptera: Coreidae), is native to Australia and a major polyphagous pest of many tropical and subtropical horticultural crops in the east and north of the country. Different plant structures (flowers, vegetative flush, and different sized fruit) of avocado, lime, and papaya crops and green bean pods (a known suitable host) were evaluated for their suitability as hosts for A. l. lutescens. Neonate to imago survivorship, the time taken to complete neonate to imago development, preovi-positional period, and fecundity were assessed for each crop. Of all the different phenological stages of the plants investigated, A. l. lutescens could complete development to imago on vegetative flush of papaya and lime, papaya flowers, and green bean pods but on no other structures tested. There was higher survivorship to the second instar when neonates fed on green bean pods or flowers or vegetative flush of avocado, lime, or papaya crops than when neonates fed on small, medium, or large fruit of these crops. Insects that developed to the imago on green bean pods were significantly heavier than insects that developed on papaya flowers or papaya vegetative flush. The mean preoviposition period was shorter, and adult females more fecund, if they completed immature development and then fed as adults on papaya vegetative flush or green beans rather than papaya flowers. The data indicate that avocado is not a suitable host for A. l. lutescens, suggesting that adult populations that cause significant pest damage to the fruit of this crop originate elsewhere.

The rednecked cane borer, Agrilus ruficollis (F.), is a pest of cultivated and wild blackberries in the midwestern and eastern parts of the United States. Damage from this pest occurs from larvae girdling primocanes and tunneling in the pith, forming galls that can potentially reduce yields. There is only one registered insecticide and no trap available for monitoring. Paints mimicking the spectral reflectance of blackberry leaves and canes of both primocane and floricane were applied to wooden dowels or corrugated plastic mimicking the shape of blackberry canes and leaves. The dowels or corrugated plastic were covered with sticky Tangletrap, and field was evaluated for attractiveness to A. ruficollis for three years, with modifications to trap design each year. Commercially available emerald ash borer, Agrilus planipennis Fairmaire, funnel traps were evaluated for attractiveness to A. ruficollis in 2014. In 2011, the greatest numbers of A. ruficollis adults were captured on prism-shaped, primocane-mimicking traps that reflected light at a peak wavelength between 540–560 nm. In 2012 and 2013, field tests demonstrated that more A. ruficollis adults were captured on green emerald ash borer traps. Testing in 2014 reinforced the previous findings that A. ruficollis was most attracted to the green emerald ash borer traps. In 2013, it was noted that colored traps usually captured significantly more A. ruficollis males than females. This indicated a need to determine if there was a chemical cue used by A. ruficollis adult females to locate and oviposit on only blackberry primocanes and not floricanes.

The Asian cockroach (Blattella asahinai Mizukubo) was introduced to Florida in 1986 and has since spread throughout the Southeastern United States. Blattella asahinai is a peridomestic pest and high population densities in residential areas can become a nuisance, especially when adults fly into homes. Few studies to date have been conducted on Asian cockroach control, and we evaluated the efficacy of Zyrox Fly Granular Bait and Maxforce Complete Granular Insect Bait against this species in the laboratory compared with the closely related German cockroach (Blattella germanica (L.)). In no-choice and two-choice assays with both species, Zyrox bait and Maxforce bait achieved nearly 100% mortality within two and five days, respectively. We also tested Zyrox bait against B. asahinai in an invasive field population in North Carolina at the label rate (2 g/m²) and at approximately three times the label rate (6.9 g/m²), and found that broadcast applications at both rates reduced populations by an average of 64 and 92%, respectively, for 35 d after the initial application. Zyrox Fly Bait appears to be effective against the Asian and German cockroaches, and could be another tool in an integrated pest management program, if its label could be extended or the active ingredient (cyantraniliprole) formulated into a cockroach bait.

Slow-acting and nonrepellent termiticides are possible candidates for nestmate to nestmate transfer called horizontal transfer. For the horizontal transfer study of spinosad, Coptotermes formosanus Shiraki was released in sand and soil at 1, 25, and 50 ppm Entrust^® for 1 h and then mixed with healthy untreated termites for 21 d at the ratio of 1:1. Donor and recipient termites began to contact and groom each other immediately after release. Mortality of termites was recorded at 1, 3, 7, and 14 d after treatment. Spinosad was more effectively transferred in sand than in soil. In sand at 25 and 50 ppm, significantly high mortality of donors and recipients was observed after 7 d. When termites were exposed to treated soil at day 21, all three concentrations resulted in significantly higher mortality compared to the control. In our laboratory study, spinosad was effectively transferred by donor termites. Transfer of spinosad depended on its bioavailability and concentration. Further study is needed to address its effects against C. formosanus under field conditions.

Bed bugs, Cimex lectularius L., are one of the most difficult urban pests to control. Pest management professionals rely heavily on insecticide sprays and dusts to control bed bugs. Dust formulations are considered to provide longer residual control than sprays. However, there are no scientific data available on the comparative efficacy of the commonly used insecticide dusts. We evaluated the efficacy of eight insecticide dust products using three exposure methods: 1) brief exposure—bed bugs crossed a 2.54-cm-wide dust-treated band, 2) forced exposure—bed bugs were continuously exposed to a dust-treated substrate, and 3) choice exposure—bed bugs were given a choice to stay on either dust-treated or untreated substrate. The brief exposure method was the most sensitive in detecting the differences among the insecticides. Only CimeXa (silica gel) dust caused 100% mortality from all three exposure methods. Other tested dusts (1% cyfluthrin, 0.05% deltamethrin, 0.075% zeta-cypermethrin + 0.15% piperonyl butoxide, 1% pyrethrins, 1% 2-phenethyl propionate + 0.4% pyrethrin, 0.25% dinotefuran + 95% diatomaceous earth, 100% diatomaceous earth) caused ≤65% mortality in a brief exposure assay. We also evaluated the horizontal transfer effect of the silica gel dust. Silica gel dust-exposed bed bugs transferred the dust horizontally to unexposed bed bugs resulting in 100% mortality at 4:6 donor: recipient ratio and 88.0 ± 5.0% mortality at 1:5 donor: recipient ratio. The results suggest silica gel is the most promising insecticide dust for controlling C. lectularius.

In the present study, the repellent effects of essential balm, a traditional medicine product in China, was tested against foraging and defending red imported fire ants, Solenopsis invicta Buren, under laboratory and field conditions. The laboratory study showed that both band- (width = 1 cm) and patch-smearing of essential balm at each concentration (0.5, 1, or 2 µl/cm²) significantly decreased the number of S. invicta foragers within the 6-h observation period. Moreover, band-smearing of 2 µl/cm² essential balm and patch-smearing of 0.5, 1, and 2 µl/ cm² essential balm inhibited most S. invicta foraging activity at 3, 6, 6, and 24 h into the experiment, respectively. The field study showed that after a disturbance was created on the S. invicta mound, there were significantly less defending ants on the substance treated (patch-smeared) with 0.5, 1, and 2 µl/cm² essential balm than the controls, but the number of ants on the substance of these three concentrations was similar. Our study suggested that essential balm is a strong repellent against foraging and defending S. invicta and could be applied when temporary protection from S. invicta is needed.

Corn fields planted with plant-incorporated Bacillus thuringiensis (Bt) proteins must have a portion of the field planted with non-Bt, isoline, plants that serve as a refuge for susceptible insects. In the Corn Belt, refuge seeds are now blended in the bag with Bt seeds for corn hybrids containing two or more toxins targeted toward the western corn rootworm, Diabrotica virgifera virgifera LeConte. Syngenta's corn hybrid, Agrisure Duracade, containing the eCry3.1Ab (event 5307) and mCry3a (event MIR604) rootworm-targeted toxins were registered as a seed blend in 2014. Western corn rootworm larval movement between the refuge plants and the Duracade plants was assessed to determine western corn rootworm survival and amount of root damage on these plants when planted in all possible seed blend scenarios. In this study, western corn rootworm larvae moved between isoline and Bt plants and adult survival was greater on Bt plants if movement from a neighboring infested isoline plant had occurred. However, root damage to these Bt plants did not reach economic levels. The low numbers of western corn rootworm larvae that did move from an infested Bt plant to an isoline plant could potentially select for resistance if they survived to adulthood.

The effect of temperature on the cotoxicity coefficient (CTC) value was used to evaluate mixture efficacy of different temperature coefficient chemicals from 15 to 35°C by exposing third-instar Apolygus lucorum (Meyer-Dür) to dip-treated asparagus bean pods. The results indicated the joint toxicity of same temperature coefficient insecticide (TCI) types were unaffected by temperature. This means that even when temperatures change, the mixture ratios of the highest CTC values remained the same, and the effect of temperature on the joint toxicity of same TCI types was only on the CTC values. However, the effect of temperature was variable when considering the joint toxicity of different TCI types. The effect of temperature on the joint toxicity of both strong positive and strong negative TCI types was clear, and the highest CTC values of mixture ratios changed with temperature regularly. When comparing the influence of temperature between strong/slight positive/negative insecticides, the results indicated a greater influence of the strong TCI. Paradoxically, the highest CTC value of the imidacloprid and methomyl mixture did not change with temperature changes consistently, even with the variance of imidacloprid ratios, a strong TCI. These results will guide pest managers in choosing the most effective insecticide mixtures for A. lucorum control under given environmental conditions.

Both an eCry3.1Ab-selected and paired control western corn rootworm, Diabrotica virgifera virgifera LeConte, colony were tested for adult longevity, egg oviposition, egg viability, and larval development in order to evaluate the potential fitness costs associated with eCry3.1Ab resistance. Adult longevity experiments were conducted by pairing virgin males and females together in plastic boxes supplied with food, water, and ovipositional medium and observed for survival time. Eggs were also collected from the ovipositional medium once a week to determine average egg oviposition and egg viability. Larval development time experiments were conducted by infesting seedling assays with 25 neonate larvae and recording larval recovery after several days. Adult longevity, average egg oviposition, and larval development time results indicated a lack of fitness costs associated with eCry3.1Ab resistance in the western corn rootworm. Results of egg viability indicated a fitness advantage for the eCry3.1Ab-selected colony with a significantly higher egg hatch than the control.

Blended refuge for transgenic plants expressing Bacillus thuringiensis (Bt) toxins has been approved in the northern United States as a resistance management strategy alternative to a structured refuge. A three-year study (2012–2014) was conducted with 54 trials across nine states in the southern United States to evaluate plant injury from lepidopteran pests of corn and yield in a corn hybrid expressing Cry1F × Cry1Ab × Vip3Aa20 (Pioneer Brand Optimum Leptra) planted as a pure stand and in refuge blends of 5, 10, and 20% in both early and late plantings. Injury by corn earworm, Helicoverpa zea Boddie (Lepidoptera: Noctuidae), and fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), was generally proportional to the percentage of non-Bt corn within each refuge blend. Across locations, ear injury in plots with 100% Cry1F × Cry1Ab × Vip3Aa20 (Optimum Leptra) corn ranged from no injury to a maximum of 0.42 cm² per ear in Mississippi in 2013. Leaf injury ratings in 100% non-Bt plots in early and late planted trials in 2014 were 86- and 70-fold greater than in 100% Cry1F × Cry1Ab × Vip3Aa20 (Optimum Leptra) plots. Plants in plots with blended refuges had significantly greater leaf injury in 2012 (5, 10, and 20% refuge blends), in the early-planted corn in 2013 (10 and 20% only), and in both early- and late-planted corn in 2014 (20% only) as compared with leaf injury in a pure stand of Cry1F × Cry1Ab × Vip3Aa20 (Optimum Leptra) seen during these years. Corn ears in plots with blended refuges also had significantly greater area of kernels injured in 2012 (5, 10, and 20%), in early- and late-planted corn in 2013 (5, 10, and 20%), and in early (10 and 20% only)- and late-planted corn (5, 10, and 20%) in 2014 as compared with ear injury in a pure stand of Cry1F × Cry1Ab × Vip3Aa20 (Optimum Leptra) seen during these years. Infestations of southwestern corn borer, Diatraea grandiosella Dyar (Lepidoptera: Crambidae), were also significantly reduced by Cry1F × Cry1Ab × Vip3Aa20 (Optimum Leptra). Despite these differences in injury, yield averaged across locations varied among refuge blends only in the late-planted trials in 2013, with greater yields in the 0% refuge blend than in the 20% blend; however, when examining yield separately by location, only two of nine locations had higher yields in the 100% Bt plots than in any of the blended refuge plots. As a complement to studying the contribution of blended refuge to delaying resistance, quantifying injury and yield in a range of refuge blends is a necessary step to provide management information on the range of lepidopteran pests that occur in the southern United States.

Western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is a serious pest of corn in the United States, and recent management of western corn rootworm has included planting of Bt corn. Beginning in 2009, western corn rootworm populations with resistance to Cry3Bb1 corn and mCry3A corn were found in Iowa and elsewhere. To date, western corn rootworm populations have remained susceptible to corn producing Bt toxin Cry34/35Ab1. In this study, we used single-plant bioassays to test field populations of western corn rootworm for resistance to Cry34/35Ab1 corn, Cry3Bb1 corn, and mCry3A corn. Bioassays included nine rootworm populations collected from fields where severe injury to Bt corn had been observed and six control populations that had never been exposed to Bt corn. We found incomplete resistance to Cry34/35Ab1 corn among field populations collected from fields where severe injury to corn producing Cry34/35Ab1, either singly or as a pyramid, had been observed. Additionally, resistance to Cry3Bb1 corn and mCry3A corn was found among the majority of populations tested. These first cases of resistance to Cry34/35Ab1 corn, and the presence of resistance to multiple Bt toxins by western corn rootworm, highlight the potential vulnerability of Bt corn to the evolution of resistance by western corn rootworm. The use of more diversified management practices, in addition to insect resistance management, likely will be essential to sustain the viability of Bt corn for management of western corn rootworm.

The Bonagota salubricola (Meyrick) is a major pest in apple orchards in Brazil, and chemical control has been the primary tool for insect management. To support the development of an insect resistance management (IRM) program, baseline studies of the susceptibility of a reference (laboratory) B. salubricola population were conducted; seven wild B. salubricola populations were monitored for susceptibility to insecticide; and the toxicity of some new chemicals to third-instar larvae and adults was evaluated by a leaf dip and ingestion bioassay, respectively. Neonates from the susceptible (laboratory) population exposed to insecticide showed an LC₅₀ ranging from 0.34 (spinetoram) to 30.19 (novaluron) µg of a.i. ml⁻¹ (88.8-fold variation), so the diagnostic concentrations for an IRM program in Brazil based on the LC₉₉ were as follows: 19.0 µg of a.i./ml chlorantraniliprole, 510.0 novaluron, 72.0 phosmet, 4.1 spinetoram, 12.8 spinosad, and 110.0 tebufenozide. Based on the LC₉₉, significant differences were not observed in the susceptibility of the field and laboratory populations to chlorantraniliprole, phosmet, spinetoram, spinosad, and tebufenozide insecticides, but there were significant differences in the survival rates of the two populations to novaluron insecticide (3.3%). All insecticides at the diagnostic concentrations showed high toxicity to third-instar larvae (mortality rates between 73 to 97%). Phosmet, spinetoram, and spinosad insecticides were toxic to B. salubricola adults (mortality >85%), while chlorantraniliprole, novaluron, and tebufenozide insecticides caused mortality below 5%. The evaluated insecticides showed high toxicity to different developmental stages of B. salubricola, so the diagnostic concentrations may be used in IRM programs in Brazil.

House dust mites (HDMs) and stored-product mites (SPMs) of various species inhabit human homes and stored agricultural products. These mites are carriers and hosts of microorganisms that enable their survival. The bacteriome from 13 species of SPMs and HDMs was analyzed and compared by 454 pyrosequencing of partial 16S rRNA gene amplicons. Altogether 128,052 sequences were obtained and assigned to 71 operational taxonomic units (OTUs) at the 97% identity level. The number of sequences in the OTUs between species of mites ranged from 6 to 31 in the individual mite species. We did not find any significant effect of diet or evolutionary origin of mites or their interaction on the composition of the mite bacteriome. In mite species with low bacterial diversity, the bacterial communities were dominated by potential symbiotic or parasitic bacteria, i.e., Cardinium in Dermatophagoides farinae (Hughes, 1961) and Aeroglyphus robustus (Banks 1906) and the enteric bacteria Erwinia in Blomia tropicalis Van Bronswijk, de Cock & Oshima, 1974 and Xenorhabdus in Tyroborus lini (Oudemans, 1924). Among the bacterial species identified, Staphylococcus, Bacillus, Kocuria, Brevibacterium, Corynebacterium, and Brachybacterium likely serve as food sources for the mites. The domestic acaridid mites carried high numbers of various bacteria that are potential threats to human health. These results contribute to the general understanding of the ecology of mite adaptation to human-made habitats.

Metcalfa pruinosa (Say, 1830) (Hemiptera: Flatidae) has caused substantial agricultural damage since its recent introduction to the Republic of Korea; however, the source of this introduction is still unclear. To examine the genetic divergence and phylogenetic relationships among several populations of M. pruinosa from Korea and foreign countries, 251 COI sequences from 251 samples collected from Korea, France, Italy, Spain, Slovenia, and the United States were newly analyzed, together with seven published COI sequences from Canada. In total, 19 haplotypes were detected from the 258 COI sequences, and three haplotypes, H1, H3, and H9, were detected from samples in Korea. The MJ network and Bayesian inference revealed that the three haplotypes of Korea were closely connected with samples of Italy, Spain, Slovenia, France, and the United States. Our study revealed the possibility of multiple invasions of M. pruinosa from Europe and/or North America into Korea.

In insects, sensory neuron membrane proteins (SNMPs) are critical peripheral olfactory proteins and highly promote the sensitivity of pheromone detection. In this study, we cloned an SNMP transcript (CpomSNMP2, GenBank KU302714) from the antennae of the codling moth Cydia pomonella (L.). Its open reading frame is 1,575 bp and it encodes a protein with 524 amino acids. CpomSNMP2 contains two putative transmembrane domains and has a large extracellular loop. Phylogenetic analysis showed that CpomSNMP2 is clustered into the group of previously characterized lepidopteron SNMP2s. Expression levels of CpomSNMP2 were significantly higher in antennae of both males and females than in tissues from the thoraxes, abdomens, legs, and wings. CpomSNMP2 was distributed in sensilla trichodea of both males and females, but only in sensilla chaetica of males. This study provides evidence for olfactory roles of CpomSNMP2 in this moth.

Screening for resistance to insect pests is one of the early stages of grass breeding programs. Pasture spittlebugs are sap-sucking insects that potentially cause severe damage to turfgrasses, including the loss of functional quality and perenniallity. The Brazilian flora has a large number of grass species with wide morphological variability and adaptability to different soil and climate conditions that can potentially be used as lawns. However, no study has screened turfgrass genotypes for resistance to spittlebug attack. In this study, we evaluated the intra- and interspecific variability of 35 turfgrass genotypes in the genera Paspalum, Axonopus, and Zoysia for resistance to the pasture spittlebugs, Deois flavopicta (Stal) and Notozulia entreriana (Berg) (Hemiptera: Cercopidae), as measured by damage scores, densities of nymphs and adults, and level of antibiosis resistance. Genotypes were grouped into three groups using cluster analysis and principal component analysis: GroupI had genotypes associated with low damage scores and high density of adult spittlebugs; GroupII had genotypes with intermediate damage scores and low density of nymphs and adults; and GroupIII was formed by genotypes with high damage scores and high nymph density. Intra- and interspecific genotypic variability was related to antibiosis resistance and morphological variation among genotypes with some indicating nonpreference resistance and others indicating tolerance resistance. Our results indicate that besides antibiosis resistance studies, it is essential to evaluate the morphological variability of grass genotypes when screening for resistance to insects. Further studies are needed to elucidate the intraspecific variability of Paspalum notatum Flüggé genotypes for resistance to spittlebug attack.

Bacillus thuringiensis (Bt) event DAS-81419-2 (Conkesta technology) in soybean, Glycine max (L.) Merrill, expresses Cry1F and Cry1Ac proteins to provide protection from feeding by several lepidopteran pests. A total of 27 field experiments across nine locations were conducted from 2011 to 2015 in southern and central Brazil to characterize the efficacy of DAS-81419-2 soybean infested with Anticarsia gemmatalis Hübner (Lepidoptera: Erebidae), Chrysodeixis includens (Walker) (Lepidoptera: Noctuidae), Heliothis virescens (F.) (Lepidoptera: Noctuidae), and Spodoptera cosmioides (Walker) (Lepidoptera: Noctuidae) during vegetative (V4) and reproductive (R2 and R4) crop developmental stages. The efficacy of DAS-81419-2 was compared to that of a non-Bt isogenic variety managed with or without applications of commercial foliar insecticides for lepidopteran control. DAS-81419-2 soybean consistently experienced defoliation levels of 0.5% or less (compared with 20.05–56.74% in the non-Bt, nonsprayed treatment) and larval survival of < 0.1% in all four species across the vegetative and reproductive plant stages evaluated. The efficacy of DAS-81419-2 was significantly higher than commercial foliar insecticides applied to the non-Bt variety. DAS-81419-2 soybeans containing two highly effective Bt proteins are expected to be a more robust IRM tool compared to single-trait Bt technologies. The consistent efficacy of DAS-81419-2 soybeans across years, locations, and crop stages suggests that it will be a valuable product for management of hard-to-control key lepidopteran pests in South American soybean production.

The cabbage aphid is a significant pest worldwide in brassica crops, including canola. This pest has shown considerable ability to develop resistance to insecticides, so these should only be applied on a “when and where needed” basis. Thus, optimized sampling plans to accurately assess cabbage aphid densities are critically important to determine the potential need for pesticide applications. In this study, we developed a spatially optimized binomial sequential sampling plan for cabbage aphids in canola fields. Based on five sampled canola fields, sampling plans were developed using 0.1, 0.2, and 0.3 proportions of plants infested as action thresholds. Average sample numbers required to make a decision ranged from 10 to 25 plants. Decreasing acceptable error from 10 to 5% was not considered practically feasible, as it substantially increased the number of samples required to reach a decision. We determined the relationship between the proportions of canola plants infested and cabbage aphid densities per plant, and proposed a spatially optimized sequential sampling plan for cabbage aphids in canola fields, in which spatial features (i.e., edge effects) and optimization of sampling effort (i.e., sequential sampling) are combined. Two forms of stratification were performed to reduce spatial variability caused by edge effects and large field sizes. Spatially optimized sampling, starting at the edge of fields, reduced spatial variability and therefore increased the accuracy of infested plant density estimates. The proposed spatially optimized sampling plan may be used to spatially target insecticide applications, resulting in cost savings, insecticide resistance mitigation, conservation of natural enemies, and reduced environmental impact.

This study was conducted at Oklahoma State University, Stillwater, OK, to evaluate the response to deltamethrin concentrations for adults of three stored-product insects, Tribolium castaneum (Herbst), Sitophilus oryzae (L.), and Rhyzopertha dominica (F.). In insect toxicological studies, knockdown is the state of intoxication and partial paralysis as a result of exposure to an insecticide. Deltamethrin concentrations ranging from 0.48 to 140 mg/m² (1 to 3,000 ppm) were sprayed on glass Petri dishes. After the dishes dried, 20 adult insects of each species were placed on the treated dishes to determine the contact toxicity of deltamethrin. Assessments for knockdown were made at 15-min intervals for up to 8 h after initial exposure and then again after 24 or 48 h. Insects were then transferred to clean untreated Petri dishes with diet and observed from 0.5 to 72 h. Mortality was assessed 72 h after transfer to untreated dishes with food material. Deltamethrin was highly effective against all three species tested and achieved 99% knockdown of insects of all species within 4 h after exposure at concentrations ≥1.2 mg/m². Although some insects recovered from initial knockdown at concentrations ≤48 mg/m², nearly all the insects were killed at 140 mg/m² when exposed for 48 h. LC₉₅ values for all species tested, for the 48-h exposure period, were <140 mg/m², the concentration of deltamethrin that could potentially be present in new ZeroFly Storage Bag fabric. ZeroFly bags are used for stored-product insect pest control.

On-farm trial with a total of 32 farmers in eight villages of Naivasha and Nakuru areas of Kenya was conducted between December 2013 and September 2014 to evaluate hermetic grain storage technologies under farmers’ management conditions. The storage technologies evaluated were metal silo and SuperGrain IV-R bag alongside the standard woven polypropylene bag with or without Actellic super dust. Moisture content, insect population, grain discoloration, and weight loss were analyzed 90, 180, and 270 d after storage. Grain moisture content remained stable over the storage period. Both metal silo and SuperGrain IV-R bag suppressed insect population, prevented grain loss and cross-infestation of insects from the surrounding environment. On the contrary, polypropylene bags allowed rapid build up of insect population and re-infestation from the surrounding environment. Grain weight losses were 1.5% in the metal silo and 1.8% in the SuperGrain IV-R bags compared to 32% in the polypropylene bags without Actellic Super dust, 270 d after storage. The present study, therefore, demonstrates that storing grains either in metal silo or SuperGrain IV-R bags would benefit farmers in reducing grain losses and improving quality. The study was of great interest to the farmers, grain storage scientists, and food security experts.

The horn fly, Haematobia irritans (L.), is a blood-sucking ectoparasite that is responsible for sizeable economic losses in livestock. The salivary gland products facilitate blood intake. Taking advantage of the identification of novel H. irritans salivary antigens (Hematobin, HTB and Irritans 5, IT5), we investigated the parasite loads, H. irritans blood intake, and antibody response of naturally infected bovines during the fly season. Fly loads and fly hemoglobin content fluctuated during the trial. Each time horn fly loads exceeded 200 flies per cattle, a reduction in horn fly blood intake was observed three weeks later. All of the cattle elicited an antibody response against HTB and IT5 that declined once the fly season was over. Cattle anti-IT5 titers were positively correlated with parasite loads and negatively correlated with fly blood intake. These results suggest that the natural changes in the H. irritans blood intake observed in this study were associated with a natural host response against horn fly salivary antigens.

A real-time PCR assay is reported for identification of Lobesia botrana (Denis and Schiffermüller) collected in California. This assay multiplexes two independent TaqMan probe systems in a single reaction tube to reduce handling time and sample exposure to environmental contaminants. One probe system targets a segment of DNA located in the internal transcribed spacer region 2 (ITS2) that is present in the L. botrana genome but absent in native North American Tortricidae. The second probe system serves as a control for DNA quality by targeting a segment of the 18S rDNA gene that is conserved in L. botrana and all of the tested nontarget species. The assay successfully diagnosed 70 Lobesia botrana specimens and 95 nontarget specimens. No false-positive or false-negative results were observed supporting its application for identification of this pest in California.