Correct gustatory recognition and selection of foods both within and outside the hive by honey bee workers are fundamental to the maintenance of colonies. The tarsal chemosensilla located on the legs of workers are sensitive to nonvolatile compounds and proposed to be involved in gustatory detection. However, little is known about the molecular mechanisms underlying the gustatory recognition of foods in honey bees. In the present study, RNA-seq was performed with RNA samples extracted from the legs of 1-, 10-, and 20-d-old workers of Apis cerana cerana Fabricius, a dominant indigenous crop pollinator with a keen perception ability for phytochemicals. A total of 124 candidate chemosensory proteins (CSPs), including 15 odorant-binding proteins (OBPs), 5 CSPs, 7 gustatory receptors (GRs), 2 sensory neuron membrane proteins (SNMPs), and 95 odorant receptors (ORs), were identified from the assembled leg transcriptome. In silico analysis of expression showed that 36 of them were differentially expressed among the three different ages of A. c. cerana workers. Overall, the genes encoding OBPs and CSPs had great but extremely variable FPKM values and thus were highly expressed in the legs of workers, whereas the genes encoding ORs, GRs, and SNMPs (except SNMP2) were expressed at low levels. Tissue-specific expression patterns indicated that two upregulated genes, AcerOBP15 and AcerCSP3, were predominately expressed in the legs of 20-d-old foragers, suggesting they may play an essential role in gustatory recognition and selection of plant nectars and pollens. This study lays a foundation for further research on the feeding preferences of honey bees.

The honey bee queen plays a central role in the Apis mellifera L. (Hymenoptera: Apidae) colony, and her high reproductive capacity is fundamental for building up the workforce of a colony. Caste development in honey bee females involves elaborate physiological pathways unleashed at the beginning of the first larval instars, with juvenile hormone (JH) playing a crucial role. Here we took advantage of established in vitro rearing techniques to conduct a 2 × 2 experimental design and test initial rearing age (young vs old) and JH treatment (JH III vs solvent control) to enlighten the role of nutrient quality and JH in shaping honey bee female fertility, morphological features related to queenliness, and key physiological parameters (hemolymph vitellogenin/Vg, sugar levels, and Vg transcript levels). Our results show that while the age at initial larval rearing had major impacts on external morphology development, where younger larvae exhibited a higher probability to develop into queen-like adults morphotypes, the JH application during the larval stage improved physiological pathways related to ovary development and metabolism during the ontogenic development. We detected that the supplementation of queen larvae with JH promoted important benefits regarding queen fertility as the increase of ovariole number and vg levels at hemolymph, both crucial factors at eggs production. The data presented here provide guidance in efforts to improve honey bee queen quality, especially in light of frequent episodes of queen failures in the beekeeping industry.

Global western honey bee, Apis mellifera (L.) (Hymenoptera: Apidae), colony declines pose a significant threat to food production worldwide. Poor nutrition resulting from habitat loss, extensive monocultures, and agricultural intensification is among the several suggested drivers for colony declines. Pollen is the primary source of protein for honey bees; therefore, both pollen abundance and diversity are critical for colony growth and survival. Many cropping systems that employ honey bee colonies for pollination may lack sufficient pollen diversity and abundance to provide optimal bee nutrition. In this observational study, we documented the diversity and relative abundance of pollen collected by honey bees in five major pollinator-dependent crops in the western United States. We sampled pollen from pollen traps installed on honey bee colonies in the following cropping systems—almond, cherry, highbush blueberry, hybrid carrot, and meadowfoam. The pollen diversity was estimated by documenting the number of different pollen pellet colors and plant taxa found in each pollen sample. The lowest pollen diversity was found in almond crop. Relatively higher quantities of pollen collection were collected in almond, cherry, and meadowfoam cropping systems. The information gleaned from this study regarding pollen diversity and abundance may help growers, land managers, and beekeepers improve pollen forage available to bees in these cropping systems.

This paper estimates the economic value of ecosystem services provided by Brazilian native bee, Xylocopa spp. Latreille (Hymenoptera: Apidae), pollination on a scale relevant to individual smallholder farmers that produce yellow passion fruit (Passiflora edulis Sims). The study areas are located in the vicinity of Pedro de Toledo and Itariri (Sao Paulo State—Brazil), in the Atlantic Forest region. The local economy is based on family farms, small stores, and ecotourism. The value was obtained using the ecological economics Avoided Cost Method, also known as replacement cost. Farms from this region informally hire temporary day laborers to supplement natural pollination with manual pollination of passion fruit flowers, so the cost of contracting temporary laborers was used to estimate the economic value of bee pollination. The value of pollination services was estimated at US$ 2,583.00 per hectare over 2 yr of P. edulis farming. Our estimates based on passion fruit farmer surveys and ecological valuation over 2 yr suggest that manual pollination accounts for 44–48% of production costs and results in a loss of ∼58% of profits when wild bee pollination services are not available and manual pollination is required. We suggest that smallholder farmers follow the suggestions of previous studies and conserve adequate forest habitat for bee nesting and foraging, plan pesticide use around flowering and pollination, and supplement bee populations to maximize the benefit of the pollination ecosystem service and profits.

Since 2016, Apis cerana sacbrood virus (AcSBV) has been recorded in Taiwan. It is epizootic in Apis cerana (Hymenoptera: Apidae) and causing serious loss of A. cerana. Herein, we performed a long-term survey of AcSBV prevalence in the populations of A. cerana in Northern Taiwan from January 2017 to July 2018. The surveillance of AcSBV prevalence in A. mellifera (Hymenoptera: Apidae) populations was starting and further confirmed by sequencing since April 2017; thus, these data were also included in this survey. In our survey, the average prevalence rates of AcSBV were 72 and 53% in A. cerana and A. mellifera, respectively, in 2017, which decreased to 45 and 27% in 2018. For the spatial analysis of AcSBV in two honey bee populations, Hsinchu showed the highest prevalence, followed by New Taipei, Yilan, Taipei, and Keelung, suggesting that AcSBV might have come from the southern part of Taiwan. Interestingly, the AcSBV prevalence rates from A. cerana and A. mellifera cocultured apiaries gradually synchronized. The result of phylogenetic analysis and comparison of the annual AcSBV prevalence in A. cerana-only, A. mellifera-only, and A. cerana/A. mellifera cocultured sample sites indicate cross-infection between A. cerana and A. mellifera; however, AcSBV may lose the advantage of virulence in A. mellifera. The evidence suggested that the transmission of AcSBV might occur among these two honey bee species in the field. Therefore, A. mellifera may serve as a guard species to monitor AcSBV in A. cerana, but the cross-infection still needs to be surveyed.

Most plant viruses maintain complex interactions with their vector or nonvector insects and can indirectly (via host plants) or directly affect the fitness of insects. However, little is known about the genes involved in the interactions between insects and transmitted or nontransmitted viruses, particularly nontransmitted viruses. Sitobion avenae (Fabricius) is a vector of barley yellow dwarf virus GAV strains (BYDV-GAV), but not a vector of wheat dwarf virus (WDV), which is transmitted by the leafhopper [Psammotettix alienus (Dahlbom)]. In this study, S. avenae was utilized to determine the transcriptomic responses after feeding on wheat infected by each of the two viruses, respectively, using an Illumina Hiseq sequencing platform. The transcriptomic data presented 61,508 genes, of which 854 differentially expressed. Moreover, in addition to sharing 208 genes, the number of differentially expressed genes (DEGs) in S. avenae exposed to BYDV was higher (800) than that when exposed to WDV (262). The DEGs related to the immune system and fitness of S. avenae in response to BYDV-/ WDV-infected plants were identified and analyzed using Gene Ontologies (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG), and the number of related DEGs was lower as nonvector than as vector. This study provides the baseline information to further examine molecular mechanisms of how wheat viruses affect S. avenae fitness and immune response either as a vector for BYDV-GAV or as a nonvector for WDV.

The spread of adventive Trissolcus japonicus (Ashmead, 1904) populations in North America is anticipated to increase biological control of Halyomorpha halys (Stål; Hemiptera: Pentatomidae), the brown marmorated stink bug. In an agricultural context, biological control will succeed if it can be integrated in an environment with insecticide applications. We investigated T. japonicus compatibility with nine conventional and organic insecticides commonly used in integrated pest management in perennial crops. Through evaluating mortality and longevity in field and laboratory trials, we determined that T. japonicus fares poorly when exposed to residues of neonicotinoids and pyrethroids. Spinosad resulted in the highest percentage of T. japonicus mortality, 100% in the laboratory and 97% in a field trial. The anthranilic diamide, chlorantraniliprole, had the lowest lethality, with no differences compared to an untreated control. Trissolcus japonicus survived insecticide applications in hazelnut orchards, and over 50% of wasps remained alive after contact with the anthranilic diamides, chlorantraniliprole and cyantraniliprole, the biopesticide Chromobacterium, and an untreated control. Our results indicate that T. japonicus is unlikely to survive and parasitize H. halys in settings that coincide with broad-spectrum insecticide application. Future T. japonicus redistributions could continue in orchards treated with anthranilic diamides and Chromobacterium. As H. halys is a landscape-level pest, orchards may also benefit from biological control if T. japonicus are released in unsprayed areas adjacent to agriculture and in urban sites.

Western flower thrips, Frankliniella occidentalis (Pergande), is a destructive insect pest in greenhouse production systems. Therefore, integrating the entomopathogenic fungus, Beauveria bassiana (Balsamo) Vuillemin, with the soil-dwelling rove beetle, Dalotia coriaria (Kraatz), targeting different aboveground and belowground life stages may help effectively manage western flower thrips populations. Two greenhouse experiments were conducted evaluating five treatments: 1) insecticides (spinosad, pyridalyl, chlorfenapyr, and abamectin), 2) B. bassiana, 3) D. coriaria, 4) B. bassiana and D. coriaria combination, and 5) water control. The estimated mean number of western flower thrips adults captured on yellow sticky cards was significantly lower for the insecticide treatment (mean range: 0–46 western flower thrips adults per yellow sticky card) than the B. bassiana and D. coriaria combination (0.3–105.1 western flower thrips per yellow card) over 8 wk. There were no significant differences in the final foliar damage ratings of chrysanthemum, Dendranthema × grandiflorum (Ramat.) Kitam., plants among the five treatments in experiment 1, but there were significant differences in experiment 2. In experiment 2, chrysanthemum plants across all treatments were not marketable due to western flower thrips feeding damage. Therefore, using B. bassiana and D. coriaria early in production should suppress population growth by targeting both foliar-feeding and soil-dwelling life stages of western flower thrips simultaneously.

The diamondback moth (DBM), Plutella xylostella L., is the most destructive pest affecting vegetable production in Kenya and around the world. Parasitoids have shown promising results in lowering the pest populations and damage caused by DBM. However, variations in host plant quality have been reported to have bottom-up effects up to the third and fourth trophic levels. We assessed the effects of two cultivated Brassica varieties (cabbage, Brassica oleracea var. capitata L. cultivar ‘Gloria F1′ and kale, B. oleracea var. acephala L. cultivar ‘Thousand headed’) on the development and performance of the specialist pest P. xylostella and two exotic parasitoids Diadegma semiclausum (Hellen) and Cotesia vestalis (Haliday). The exposed larval period of DBM took about 1.5 d longer on kale than cabbage and the total immature development time of both females and males was significantly longer on kale than cabbage. Higher pupal weight and higher fecundity were recorded on DBM fed on kale. Development time of D. semiclausum and C. vestalis was not affected by the host crop as was the parasitism rate of D. semiclausum. Heavier male pupae and larger adults of D. semiclausum, as well as more fecund adults of C. vestalis, were obtained from hosts fed on cabbage. Larger adults of C. vestalis were obtained from herbivores fed on kale. These results show potentially positive effects of host plant allelochemicals that are detrimental to herbivores while promoting parasitoid development and performance, which can be harnessed for the control of DBM.

Schizotetranychus oryzae Rossi de Simons attains pest level in rice crops. This study aimed to evaluate the biological features of Neoseiulus californicus (McGregor) feeding on S. oryzae on leaves of flooded rice. We also evaluated N. californicus behavior in response to S. oryzae and conspecific cues. Thirty-two eggs were individually placed in arenas on rice leaves and fed on S. oryzae for the biological test.Three cultivars were used to test the behavior of N. californicus in response to S. oryzae and conspecific cues. The intrinsic rate of increase (rm) of the predator feeding on S. oryzae was 0.21 female/female/day. Schizotetranychus oryzae was considered a suitable prey for N. californicus under laboratory conditions. The results obtained in behavioral tests showed that N. californicus preferred laying eggs on leaves uninfested by S. oryzae and it did not show oviposition preference in arenas with conspecific predator cues. This predator was found in all rice cultivation areas in the state of Rio Grande do Sul, Brazil. Neoseiulus californicus proved to be a control agent for S. oryzae.

We studied the direct (lethal) and indirect (sublethal) effects of field-aged insecticide residues of spinetoram, chlorantraniliprole and lambda-cyhalothrin on adults and larvae of the green lacewing, Chrysoperla johnsoni Henry, Wells and Pupedis an important generalist predator in western United States pear orchards. We applied formulated pesticides using their high label rates mixed with 935 liters/ha of water on pear trees utilizing an airblast sprayer. We collected leaves from the experiment trees at 1, 7, 14, 21 days after treatment (DAT) and exposed C. johnsoni adults and second instars to insecticide residues or untreated control for 72 h. We monitored larvae that survived the treatments until adult emergence (for larvae) or adults until death (for paired adults). We assessed the adults for acute and chronic mortality, longevity, fecundity, and fertility, and larvae for acute mortality, sex ratio, developmental time, and survival of adults emerged from treated second instars, to determine lethal and sublethal effects. The field-exposed insecticide residues of chlorantraniliprole exhibited higher lethal and sublethal toxicities on C. johnsoni during the 21-d leaf collection period when compared with the field-exposed residue toxicities of lambda-cyhalothrin and spinetoram. The field-exposed residue of lambda-cyhalothrin and spinetoram exhibited moderate toxicities on C. johnsoni during the 21-d leaf collection period. Results from this study show similar trends with previous studies using multiple routes of exposure laboratory assays and field experiments on Chrysoperla sp. The direct and indirect effects of field-aged residues of spinetoram, chlorantraniliprole, and lambda-cyhalothrin on C. johnsoni are discussed.

Aspects of the nutritional ecology and life histories of five predatory coccinellids (Coleoptera: Coccinellidae)—two arboreal predator species, Oenopia conglobata contaminata (Menetries) and Adalia bipunctata (Linnaeus), and three herbaceous dwelling predator species, Coccinella undecimpunctata aegyptica (Reiche), Exochomus nigripennis (Erichson), and Hippodamia variegata (Goeze) (Coleoptera: Coccinellidae)—were compared when fed either Agonoscena pistaciae Burckhardt and Lauterer (Hemiptera: Aphalaridae), the key psylla pest of pistachio trees, or Aphis gossypii Glover (Hemiptera: Aphididae), a common aphid on herbaceous plants in pistachio orchards.The putative habitat preferences of four of the five coccinellids studied were consistent with their performance on the major herbivore present in those two habitats in terms of net reproductive rate. Oenopia conglobata contaminata and Ad. bipunctata showed higher net reproductive rates when fed on pistachio psylla prey, whereas C. undecimpunctata aegyptiaca and E. nigripennis had better reproductive output on aphid prey. Moreover, E. nigripennis was the most specialized of these generalist predators, eating more and reproducing better when fed aphids. Hippodamia variegata was relatively unaffected by the diet offered, suggesting greater ability to switch among herbivore prey and perhaps better potential as a biocontrol agent with abilities to exploit Ag. pistaciae without large life history tradeoffs.

The emerald ash borer (EAB), Agrilus planipennis Fairmaire, a buprestid beetle native to Asia, has become a serious pest of ash trees (Fraxinus spp.) in North America since the early 2000s. Due to the impracticality of applying insecticides in natural forests, biocontrol is the most viable method to manage EAB in natural ecosystems. Here, we report the first evidence for the establishment and impact of Spathius galinae Belokobylskij & Strazenac, a larval parasitoid first released in North America in 2016 and 2017 at six mixed-hardwood forest sites, in Connecticut, New York, and Massachusetts. We also report current levels of abundance and parasitism of another introduced larval EAB parasitoid, Tetrastichus planipennisi Yang (Hymenoptera: Eulophidae), released in 2015 and 2016 in these same sites. Spathius galinae was recovered at all release sites in 2018, and its density in sampled trees had increased 1.5- to 20-fold (relative to the first postrelease sample year), reaching a final density of 2.3–14.3 broods/m² of phloem area and causing 13.1–49.2% marginal rate of parasitism at four of the six sites. In contrast, T. planipennisi was only recovered in 2018 at four of the six release sites, and both its density (0.1–2.3 broods/m² of phloem area) and parasitism (0.1–5.6%) were lower than that of S. galinae throughout the study at the four sites where recoveries were made. Our data fill a critical gap in the development of a biocontrol-based EAB management plan to protect surviving ash trees capable of reaching maturity and producing replacement trees.

The amaranth leaf-webber, Spoladea recurvalis (Fabricius; Lepidoptera: Crambidae), is a serious pest of Amaranthus sp. in Africa and Asia. Apanteles hemara (Nixon; Hymenoptera: Braconidae) is by far the most important larval endoparasitoid of the amaranth leaf-webber. We examined the effects of host density and age on the biological characteristics of A. hemara. The regression model of the number of hosts supplied to A. hemara against the number of larvae parasitized resulted in a curve corresponding to type II functional response, with a significant increase in the number of hosts parasitized up to the density of 30 hosts before being constant up to 40 hosts. In contrast, the parasitism rate decreased linearly with increasing host densities. Development time, sex ratio, and adult longevity were not significantly affected by host density. The immature parasitoid mortality was significantly higher at higher host densities. Apanteles hemara did not parasitize 7-d-old larvae and beyond, while parasitism was significantly higher among 1- to 2-d-old compared with 3- to 4-d-old larvae. Immature parasitoid mortality was 2.6 times higher in 1- to 2-d-old larvae compared with 5- to 6-d-old larvae. The developmental period of the parasitoid from egg to adult was longest among 1- to 2-d-old larvae and least among 5- to 6-d-old larvae. Nonreproductive mortality was markedly higher among 1- to 2-d-old larvae compared with the older larvae. Adult female A. hemara were significantly larger on 3- to 4-d-old larvae compared with either 1- to 2-d-old or 5- to 6-d-old larvae. We discuss the implications of our results for the interpretation of functional response in parasitoids, mass rearing, conservation, and augmentative biological control of S. recurvalis.

Most adult parasitoids depend on carbohydrate-rich food as an energy source for longevity, fecundity, and mobility. Thus, providing sugars has been proposed as a technique to maximize the biological control efficacy of parasitoids. However, the sugars provided for parasitoids need to be carefully selected because herbivore hosts might also benefit. Here we explore the effects of 12 naturally occurring sugars on the longevity and fecundity of the rice pest, Chilo suppressalis, and the longevity of its parasitoid, Cotesia chilonis, as well as the effect of sugars on sugar consumption of C. chilonis. Results showed that none of the tested sugars significantly impacted the longevity of C. suppressalis, but fructose, glucose, maltose, and sucrose significantly increased its fecundity. The longevity of C. chilonis was significantly increased when fed fructose, glucose, sucrose, maltose, trehalose, and melezitose. Our data suggest that C. chilonis consumed larger quantities of glucose, fructose, and sucrose followed by maltose, melezitose, and trehalose and the longevity of C. chilonis was positively correlated to sugar consumption. Our data also suggest that the herbivore C. suppressalis and its parasitoid C. chilonis responded differently to trehalose and melezitose. Although additional studies are needed, our data suggest that these sugars could be considered as candidate components for sugar sprays to enhance the activity and efficacy of C. chilonis, but without benefiting its pest host.

The fumigation activity of phosphine (PH₃) and ethyl formate (EF) and their phytotoxicity to 13 imported foliage nursery plant species were evaluated. The lethal concentration and time (LCT₉₉) values of the PH₃ indicated that the susceptibility of the nymphs (3.95 and <0.45 mg·h/liter, respectively) was higher than that of the adults (5.29 and 3.66 mg·h/liter, respectively) of two mealybugs [Pseudococcus longispinus (Targioni-Tozzetti) and P. orchidicolaTakahashi]. The highest concentration reduction rate of PH₃ and EF on the 13 foliage nursery plants in the 12-liter desiccator was 41.5% for Heteropanax fragrans and 71.7% for Schefflera arboricola, respectively, which indicates that PH₃ has a lower sorption rate than EF. The phytotoxicities of PH₃-treated foliage nursery plants did not significantly differ from those of the nontreated plants, but EF caused phytotoxicity in 11 foliage nursery plants a week after treatment. When the exposure time of PH₃ increased to 24 h, the adults and nymphs of both mealybug species showed 100% mortality in the 0.5 m³ fumigation chamber. In the 10 m³ fumigation container used in the field, there was 100% mortality of both mealybugs after treatment with 2 g/m³ PH₃ for 24 h at 16°C. These results indicate that EF is not a suitable mealybug fumigant due to its high sorption and phytotoxicity to foliage nursery plants, despite fumigation activity against the two species. However, PH₃ seems to be suitable for mealybug fumigation in foliage nursery plants and can be used as a substitute for methyl bromide.

The edible katydid, Ruspolia differens (Serville), is an economically and nutritionally important insect species that is common and widespread in Africa. Currently, there are aims to develop mass-rearing methods suitable for this species, but a critical first step would require a suitable oviposition medium to ensure economic and easy production of eggs with high hatchability. The objective of the present study was to determine the oviposition preference and egg hatchability in different oviposition media. Based on preliminary testing of a wide range of different materials, two oviposition media, namely, artificial (folded plastic cloth) and natural medium (leaf sheath of Pennisetum purpureum Schumach attached to a stem), were selected for a detailed experiment. A two-choice laboratory experiment was conducted in which R. differens females were able to select between the folded plastic cloth (one side covered with short fleece) and a leaf sheath of P. purpureum (wrapped in polyester fiber). The results show that R. differens laid almost three times more eggs on the plastic cloth (mean ± SE, 169.3 ± 18.0) than on the natural substrate, P. purpurem (69.0 ± 13.9). However, the hatchability (overall mean% ± SE, 56.5 ± 4.7) of eggs in the two media did not differ. Folded plastic cloth, therefore, provided a more effective medium for egg laying by R. differens, and is thus a promising oviposition medium for getting natural populations to start laying eggs in future mass rearing.

Bradysia odoriphaga Yang & Zhang (Diptera: Sciaridae) is an important pest of Chinese chives. Information on the effects of biotic and abiotic factors on the flight performance of B. odoriphaga is crucial for understanding the pest's ability to disperse and migrate. In this study, the effects of sex and air temperature on the flight performance of B. odoriphaga imagoes were assessed by tethering individual imagoes to computerized flight mills for a 10-h experiment. The results showed that the percentage of imagoes that flew a particular distance gradually decreased as flight distance increased. The percentage of imagoes was significantly higher for males than females when the flight distance was <300 m. Sex and air temperature significantly affected average flight time (which ranged from 14.6 to 68.3 min) and average flight distance (which ranged from 10.4 to 107.2 m), but did not significantly affect average flight speed (which ranged from 3.8 to 6.4 m/min). For both females and males, the average flight distance and flight time were shortest at 18°C and longest at 22°C; the interaction between air temperature and sex was not significant.The results suggest that B. odoriphaga has a poor potential for long-distance migration.These findings will be helpful for developing forecasting and management systems for B. odoriphaga.

Essential oils are well known to act as biopesticides. This research evaluated the acute toxicity and synergistic effect of essential oil compounds in combination with cypermethrin against Spodoptera litura Fabricius (Lepidoptera: Noctuidae). The effects of distillation extracts of essential oils from Alpinia galanga Zingiberaceae (Zingiberales) rhizomes and Ocimum basilicum Lamiaceae (Lamiales) leaves; one of their primary essential oil compounds 1,8-cineole; and linalool were studied on second-instar S. litura by topical application under laboratory conditions. The results showed that A. galanga had the highest control efficiency, whereas1,8-cineole provided a moderate efficacy. The mixtures of linalool, 1,8-cineole, O. basilicum, or A. galanga with cypermethrin were synergistic on mortality. Activity measurements of the main detoxification enzymes show that linalool and 1,8-cineole inhibit the activity of cytochromes P450 and carboxylesterases, which could explain their synergistic effect. Based on our results, the use of these mixtures represents an ideal eco-friendly approach, helping to manage cypermethrin resistance of S. litura.

To further develop integrated pest management (IPM) strategies against two sucking insect pests, Aleurocanthus spiniferus (Quaintanca) (Hemiptera: Aleyrodidae) and Toxoptera aurantii Boyer (Hemiptera: Aphididae), in Chinese tea plantations, it is important to evaluate the effects of insecticides on biological control agents, such as the seven-spot lady beetle, Coccinella septempunctata L. (Coleoptera: Coccinellidae). Therefore, we tested the toxicity and sublethal effects of spirotetramat, clothianidin, lambda-cyhalothrin, and bifenthrin on C. septempunctata.The side effects of the active ingredients of the insecticides were evaluated with residual contact tests on the larvae of C. septempunctata in the laboratory. Spirotetramat and clothianidin exhibited lower lethality to C. septempunctata as shown by the higher LC₅₀ values and had higher selectivity for A. spiniferus and T. aurantii based on the selective toxicity ratios being superior to other tested insecticides. Spirotetramat also did not affect survival, longevity, fecundity, and egg hatching of C. septempunctata. Clothianidin and bifenthrin prolonged the duration of larval development stages of C. septempunctata obviously. Clothianidin at >2.5 mg/liter, lambda-cyhalothrin at >0.03 mg/liter, and bifenthrin at >0.125 mg/liter significantly reduced the survival and pupation rates of C. septempunctata larvae. Furthermore, spirotetramat at 3.125 mg/liter was harmless (IOBC class 1) to larvae of this coccinellid species. Based on the results, spirotetramat was innocuous to C. septempunctata larvae compared with clothianidin, lambda-cyhalothrin, and bifenthrin. Therefore, spirotetramat might be incorporated into IPM programs in combination with C. septempunctata for control of A. spiniferus and T. aurantii in the tea plantations.

Two-spotted spider mite, Tetranychus urticae Koch (Tetranychidae: Acariformes), is one of the most important agricultural pests in the world. Their populations have a tendency of rapidly developing resistance to acaricides, making it necessary to have a variety of active ingredients for sustainable chemical control of this pest. We investigated acaricidal properties of a relatively new insecticide spirotetramat using its commercial formulation, Movento Energy. Spirotetramat applied at concentrations equivalent to the field rates of 78 and 90 g ai/ ha had a strong negative effect on the survival of the treated T. urticae, with ca. 95% of females and ca. 65% of males dying after the treatment. Spirotetramat appeared to interfere with female reproductive system. Many of the dead treated females had eggs stuck in the oviduct and protruding from their bodies. Surviving treated females did not lay eggs. Furthermore, untreated females that mated with treated males did not produce female offspring and displayed the symptoms of spirotetramat poisoning. Toxic effects continued manifesting themselves after female mites were transferred from treated onto untreated plant culture. Contrary to previous studies, contact toxicity was also detected. None of the treated immature stages survived to adulthood. Based on these results, spirotetramat may be a good option for integrated pest management in crops that are simultaneously affected by sucking insects and spider mites and in pesticide rotation sequences that are a part of integrated resistance management programs.

Corn earworm, Helicoverpa zea (Boddie) Lepidoptera: Noctuidae, has not been considered an economic pest of field corn. Historical losses estimates ranged from 1.5 to 2.5%, and a large number of foliar insecticide applications would be needed to minimize infestations. In recent years, Bt, Bacillus thuringiensis (Berliner) field corn, Zea mays (L.) Poales: Poaceae, technologies that exhibit activity against corn earworm have been introduced. However, it is unclear how much damage to corn ears (number of damaged kernels) is required to reduce yield. In this study manual damage methods were utilized to inflict defined levels of kernel damage and to impose damage at levels greater than observed with natural corn earworm infestations. Bt corn hybrids expressing the Agrisure Viptera (Vip 3A) trait were used to minimize injury from natural infestations of corn earworm. Manual kernel damage was imposed at R3 stage to mimic corn earworm feeding while avoiding interference with pollination.These methods were used in experiments where treatments were applied to individual ears and hand-harvested and in experiments where treatments were applied to all primary ears in the plot and machine-harvested. Damage of ≥60 kernels per ear was required to significantly reduce yield regardless of harvest method. Kernel damage from natural corn earworm infestations reported in other studies was much lower than 60 kernels per ear. Timely planting is a key component of all integrated pest management programs. Field corn planted during the recommended planting window for optimum yield is unlikely to experience corn earworm damage great enough to reduce yield.

Six trap designs were assessed for capturing noctuid moths in field trials in the United Kingdom and Argentina. The traps were baited with either a sex pheromone for Autographa gamma in the UK trials and Helicoverpa gelotopoeon (Dyar) (Lepidoptera: Noctuidae) in Argentina, or a floral odor blend. In the UK trials the Universal Trap and a funnel sleeve trap were compared; in Argentina the funnel sleeve trap, a homemade bucket trap, and (L.) (Lepidoptera: Noctuidae) three sticky traps: LepTrap, wing trap, and delta trap were compared. Comparisons were made between the traps and attractants and captures of noctuid moths and nontarget insects. Traps baited with the floral attractant caught a lower number but a wider range of noctuid species including Helicoverpa, Spodoptera, Rachiplusia, Dargida, Mythimna, Chrysodeixis, Agrotis, and Autographa spp., and nontarget insects. In the UK trials, the funnel sleeve trap caught significantly more A. gamma than the Universal Trap. The addition of an insecticide to the Universal Trap marginally increased trap catches. In the Argentinian trials, the homemade bucket and the sleeve traps outperformed all sticky traps in most situations regardless of crop environment and attractant type. The homemade bucket and the funnel sleeve traps are also considerably cheaper than the other trap designs. Of the sticky traps the LepTrap caught more noctuids than the wing and delta traps. The results suggest that the bucket trap and the sleeve trap have a much greater maximum capacity and possibly a greater retention efficiency for noctuid moths compared to the sticky traps.

Reverse transcription quantitative polymerase chain reaction (PCR) has become an invaluable technique for analyzing gene expression in many insects. However, this approach requires the use of stable reference genes to normalize the data. Chlorops oryzae causes significant economic damage to rice crops throughout Asia. The lack of suitable reference genes has hindered research on the molecular mechanisms underlying many physiological processes of this species. In this study, we used quantitative real-time PCR to evaluate the expression of eight C. oryzae housekeeping genes glyceraldehyde-3-phosphate dehydrogenase (GAPDH), β-actin (βACT), beta-tubulin (βTUB), Delta Elongation factor-1 (EF1δ), ribosomal protein S11 (RPS11), RPS15, C-terminal-Binding Protein (CtBP), and ribosomal protein 49 (RP49) in different developmental stages and tissues in both larvae and adults. We analyzed the data with four different software packages: geNorm, NormFinder, BestKeeper, and RefFinder and compared the results obtained with each method. The results indicate that PRS15 and RP49 can be used as stable reference genes for quantifying gene expression in different developmental stages and larval tissues. GAPDH and βACT, which have been considered stable reference genes by previous studies, were the least stable of the candidate genes with respect to larval tissues. GAPDH was, however, the most stable reference gene for adult tissues. We verified the candidate reference genes identified and found that the expression levels of Cadherins (Cads) changed when different reference genes were used to normalize gene expression. This study provides a valuable foundation for future research on gene function, and investigating the molecular basis of physiological processes, in C. oryzae.

The bermudagrass stem maggot, Atherigona reversura Villeneuve (Diptera: Muscidae), was first reported damaging bermudagrass Cynodon dactylon (L.) Pers grown for forage in 2010 in the southeastern United States. Injury results from individual larvae feeding internally on the vascular tissue just above the terminal node of the grass stem. Injury slows plant growth and reduces forage accumulation. To address the need for economic guidelines to manage this new pest, the relationship between the percent of stems damaged by bermudagrass stem maggot and forage yield was measured in commercial bermudagrass hay fields in northcentral Texas. Yield loss was estimated to be 9.97 kg/ha (8.90 lbs /acre) for each percentage of stems with bermudagrass stem maggot damage.This relationship was used to calculate economic injury levels for a range of hay market values and control costs. The impact of stem damage on protein content, energy, and digestibility of bermudagrass hay was also investigated. Although there was a significant trend for declining forage quality with increasing stem damage, stem damage explained very little of the model's variability.

Immature stages of the boll weevil, Anthonomus grandis grandis Boh. (Coleoptera: Curculionidae), develop protected inside cotton fruiting structures. Therefore, the adult beetles have become the main target of insecticide applications. The use of insect growth regulators (IGRs) is recommended against immatures, even though they may also affect the survival and reproductive traits of adult insects. The present study evaluated the impact of a juvenile hormone analog (pyriproxyfen), an ecdysteroid agonist (methoxyfenozide), and a chitin biosynthesis inhibitor (lufenuron) on adult cotton boll weevils, a key cotton pest. Mated and virgin beetles were treated by feeding them contaminated squares and cotton leaf discs that were previously immersed into pyriproxyfen, methoxyfenozide, and lufenuron solutions at field-rate concentrations. After exposure, treated couples were caged onto cotton plants, and survival, fecundity, and egg viability were evaluated. The IGRs neither affected the survival nor fecundity of adult boll weevils. On the other hand, egg viability was significantly reduced by lufenuron, regardless of whether the females were treated premating or postmating or whether their pairs were either treated or untreated. However, egg viability increased as the females aged since the initial exposure date to lufenuron, indicating a potential transovarial effect of this insecticide. Our results indicate that pyriproxyfen and methoxyfenozide do not affect adult boll weevils, whereas lufenuron temporarily reduces the egg viability of this key cotton pest.

Thaumetopoea pityocampa (Denis and Schiffermüller) is a severe defoliator of various species of Pinus and Cedrus, while the urticanting hairs produced by its larvae cause public health problems for humans and pets. In the present study, we report results of trials (from summer 2015 until winter 2017) of mating disruption for management of T. pityocampa in different areas of Italy and Greece. Overall, the total number of male moths captured in mating disruption-treated plots over each season (70) was significantly lower than the respective number in untreated plots (780). The total number of winter nests was likewise significantly less in the mating disruption plots (13) compared with control plots (147). Our results indicate that mating disruption can be an important tool for judicious, insecticide-free control of T. pityocampa in urban, suburban, and recreational areas, where many alternative control measures are not available.

We present research on the chemical ecology of 14 species of longhorned beetles (Coleoptera: Cerambycidae), in four tribes of the subfamily Cerambycinae, conducted in east-central Illinois over 8 yr. Adult males produce aggregation-sex pheromones that attract both sexes. Twenty independent field bioassays explored the pheromone chemistry of the species and tested the possible attractive or antagonistic effects of compounds that are not produced by a given species, but are pheromone components of other species. Analyses of beetle-produced volatiles revealed compounds that had not been reported previously from several of the species. The most common pheromone component was (R)-3-hydroxyhexan-2-one, but pheromones of some species included isomers of the related 2,3-hexanediols. Males of the congeners Phymatodes amoenus (Say) and Phymatodes testaceus (L.) produced pure (R)-2-methylbutan-1-ol. Enantiomers of 2-methylbutan-1-ol also proved to be powerful synergists for Megacyllene caryae (Gahan), Sarosesthes fulminans (F.), and Xylotrechus colonus (F.). The major components of pheromone blends were consistently present in collections of headspace volatiles from male beetles, and only the major components were inherently attractive to a subset of species when tested as single components. Minor components of some species acted as powerful synergists, but in other cases appeared not to influence attraction. Among the minor components identified in headspace extracts from males, 2,3-hexanedione and 2-hydroxyhexan-3-one appeared to be analytical artifacts or biosynthetic by-products, and were neither attractants nor synergists. The antagonistic effects of minor compounds produced by heterospecific males suggest that these compounds serve to maintain prezygotic reproductive isolation among some species that share pheromone components.

We tested 3-methyl-2-cyclohexen-1-one (MCH) and novel semiochemicals as potential spruce beetle (Dendroctonus rufipennis Kirby) (Coleoptera: Curculionidae, Scolytinae) repellents over multiple years in Utah and Colorado trapping bioassays. MCH is a known spruce beetle repellent and our testing revealed Acer kairomone blend (AKB) and isophorone plus sulcatone as repellents. We subsequently tested these semiochemicals for area and single tree protection to prevent spruce beetle attacks at locations in Utah, Colorado, Wyoming, New Mexico, and Alaska. Individual tree protection trials found MCH–AKB provided significant protection against spruce beetle attacks in the southern Rocky Mountains but not in Alaska. Adding sulcatone or doubling MCH–AKB pouches did not further enhance protection. A degree of protection was extended to spruce at least 10 m distant from the repellents, including in Alaska. Tree diameter was not a significant covariate among treated trees but was positively correlated with the probability of infestation for surrounding spruce. In area protection trials, spruce in control plots were 2.4 times more likely to be in a higher severity attack class compared with spruce in plots treated with MCH–AKB pouches deployed at 30 sets per hectare. Tree diameter had a significant, positive relationship to the probability of infestation. We found MCH–AKB to offer a high degree of protection against beetle attack in Engelmann spruce (Picea engelmannii Parry ex Engelm.) (Pinales: Pinaceae) (Picea engelmannii Parry ex Engelm.) (Pinales: Pinaceae), especially for single tree protection (66% of control trees were strip- or mass-attacked compared with 6% of repellent-treated trees). AKB requires registration and labeling, however, before this economical and environmentally benign semiochemical can be used operationally.

Neoseiulus fallacis (Garman) is a predatory mite that is common in apple orchards and distributed throughout North America. However, N. fallacis may be susceptible to pesticides used for the management of crop pests. This study aimed to evaluate the temporal effects of commonly used insecticides on N. fallacis survival. Neoseiulus fallacis adults were exposed to field-aged residues, and mortality and lethal time were measured over 96 h of exposure. Carbaryl caused high mortality to N. fallacis and the shortest lethal time values (LT₅₀), followed by spinetoram, with moderate lethal time values. Esfenvalerate, acetamiprid, chlorantraniliprole, and novaluron showed little to no lethality to N. fallacis following exposure to dry field-aged residues. The results of this study provide important field-relevant knowledge that is often void from laboratory-based studies, which can aid integrated pest management (IPM) decision-makers in apple production systems.

One of the major challenges facing citrus integrated pest management (IPM) in California is the recent, sharp increase in the acreage of mandarins being planted. The current citrus IPM guidelines have been established from years of experiments and experience in oranges, with no specific guidelines for mandarins. In the absence of research into key arthropod pest effects in mandarins, the assumption that the pest management practices for oranges appropriately transfer for optimal production in mandarins has not been tested. We used a data mining or ‘ecoinformatics' approach in which we compiled and analyzed production records collected by growers and pest control advisors to gain an overview of direct pest densities and their relationships with fruit damage for 202 commercial groves, each surveyed for 1–10 yr in the main production region of California. Pest densities were different among four commonly grown species of citrus marketed as mandarins (Citrus reticulata, C. clementina, C. unshiu, and C. tangelo) compared with the standard Citrus sinensis sweet oranges, for fork-tailed bush katydids (Scudderia furcata Brunner von Wattenwyl [Orthoptera: Tettigoniidae]), and citrus thrips (Scirtothrips citri Moulton [Thysanoptera: Thripidae]). Citrus reticulata had notably low levels of fruit damage, suggesting they have natural resistance to direct pests, especially fork-tailed bush katydids. These results suggest that mandarin-specific research and recommendations would improve citrus IPM. More broadly, this is an example of how an ecoinformatics approach can serve as a complement to traditional experimental methods to raise new and unexpected hypotheses that expand our understanding of agricultural systems.

Sterile insect technique (SIT) for Queensland fruit fly, Bactrocera tryoni Froggatt, Australia's most economically damaging fruit fly species, is currently undergoing a major renewal and expansion. SIT relies on efficient and economical mass-rearing procedures that produce high-quality flies. Two solid larval diets, carrot and lucerne chaff, have traditionally been used to rear Queensland fruit fly. Recently, a gel larval diet has been developed to eliminate biological bulking agents from the mass-rearing process, but to date, there has been no direct comparison of gel larval diet with traditional solid diets. In the present study, the performance of flies reared on gel larval diet was compared with the performance of flies reared on carrot and lucerne chaff diets. In addition, to investigate whether the performance of reared flies depends on ancestral diet as well as tested diet, we sourced eggs from a colony maintained on carrot diet and from a colony maintained on a lucerne chaff diet. Overall, the gel diet was as good or better than the solid diets in all quality control parameters, including, egg–larval duration, pupal number, pupal recovery, adult emergence, percentage of fliers, and rate of fliers. Of note, larvae developed faster and pupated more synchronously on the gel diet than on either of the solid diets. At the loading densities used, gel and carrot diets produced less waste than lucerne chaff diet. Gel diets offer a rearing solution for Queensland fruit fly that eliminates biological bulking agents and yields faster and more synchronous larval development without compromising productivity or quality.

Spotted-wing drosophila, Drosophila suzukii (Matsumura), an economically damaging invasive species of numerous fruit crops, was first detected in Minnesota in 2012. High fecundity, and short generation times facilitated a rapid rise in the global pest status of D. suzukii, particularly in North America and Europe. To date, the majority of crop injury research has focused on fruit crops such as blueberries, raspberries, and cherries. However, little is known regarding the impact of D. suzukii on the wine grape industry in the upper Midwest region of the United States. Field trials were conducted in Minnesota during the summers of 2017–2018 to examine season-long phenology of D. suzukii in wine grape vineyards and wineries, and to assess the efficacy of exclusion netting for control of D. suzukii. Four treatments were evaluated, 1) open plot check (control), 2) open plot treated with an insecticide, 3) exclusion netting, and 4) exclusion netting, with artificial infestations of D. suzukii adults. Exclusion netting was applied at véraison and removed at harvest. On each sample date, 20 berries (10 intact and 10 injured) were collected from each plot for dissection. The number of larvae and adults were recorded for each berry to determine infestation levels. As shown by mean larval infestations and injured berries across treatments, exclusion netting provided a significant reduction in the level of D. suzukii infested berries when compared with the untreated check. These results indicate that exclusion netting could provide an effective alternative management strategy for D. suzukii in wine grapes.

Despite limited efficacy data, do-it-yourself (DIY) insecticide products often promise low-cost alternatives to professional pest control. Total release foggers (TRFs, ‘bug bombs’), which are prominent DIY products, were recently shown to be ineffective at reducing German cockroach (Blattella germanica L.) infestations, in contrast to highly effective baits. However, the reason(s) for TRF failure remain unknown. Therefore, we investigated insecticide resistance of apartment-collected cockroaches from homes where TRFs failed. In topical (direct) application assays, resistance to cypermethrin (a common active ingredient in TRFs) was 202 ± 33 times that of a laboratory insecticide-susceptible population (based on LD₅₀ ratios), while resistance to fipronil, a common bait active ingredient, was considerably lower at 14 ± 2 times that of the laboratory insecticide-susceptible population. The addition of PBO, a P450 inhibitor that synergizes pyrethroids, enhanced the efficacy of cypermethrin, but only at high doses of cypermethrin. Additionally, >96% of screened cockroaches possessed at least one copy of the L993F mutation in the voltage-gated sodium channel, known to confer resistance to pyrethroids (knockdown resistance, kdr). Because TRF treatments killed insecticide-susceptible sentinel cockroaches but failed to kill apartment-collected cockroaches, these results suggest that pyrethroid resistance is a major factor contributing to the failure of TRFs. Multiple mechanisms of resistance, including metabolic detoxification of the pyrethroids and kdr mutations that confer target-site insensitivity, suggest that TRFs would lack efficacy against German cockroaches in residential settings, where high levels of pyrethroid resistance have been documented globally.

The German cockroach Blattella germanica (L.) (Blattodea, Ectobiidae) is one of the most common indoor pests in multifamily housing communities. Our objectives were to investigate the spatial distribution patterns of German cockroach infestations in a multiunit dwelling before and after building-wide integrated pest management (IPM) implementation. The study was conducted in a high-rise apartment building with 188 residential units in Paterson, New Jersey. All the identified infestations were treated by researchers using IPM implementation, and monitoring was repeated at 6 and 12 mo after the initial survey. We examined cockroach spatial distribution patterns within and between apartments. Traps placed by the stove and refrigerator caught significantly more cockroaches than traps placed in the kitchen cabinet under the sink or in the bathroom. At month 0, cockroach infestations within the building were spatially correlated. At 12 mo after implementation of IPM, only infestations cross the floors remained to be correlated. Therefore, IPM not only eliminates cockroach infestations in individual units, but also reduces cockroach dispersal between units.

Laboratory studies of Coptotermes formosanus Shiraki (Blattodea: Rhinotermitidae) often employ the use of field-collected foraging populations of individuals as defined colonies. The biological relevance of this practice is often called into question, because these colonies lack a full composition of reproductive castes and brood, which may have physiological and behavioral consequences. Rearing intact laboratory colonies can be done; however, it is time-consuming and labor-intensive. The artificial fusion of field-collected foraging populations with a young, laboratory-reared incipient colony may provide whole, intact colonies for laboratory research. The current study measures survivorship of fused colonies using laboratory-reared complete incipient colonies ranging in age from 0 to 5 mo, fused with 100 workers and 10 soldiers from field-collected populations of different colonial origin. Results indicate that 60% of colony fusion was successful when the incipient colony introduced is 5 mo of age. This method of colony fusion will provide researchers with intact colonies using minimal resources.

Organisms live in complex multivariate environments. In agroecosystems, this complexity is often human-induced as pest individuals can be exposed to many xenobiotics simultaneously. Predicting the effects of multiple stressors can be problematic, as two or more stressors can have interactive effects. Our objective was to investigate whether indirect glyphosate-based herbicide (GBH) exposure of the host plant has interactive effects in combination with an insecticide (azinphos-methyl) on an invasive pest Colorado potato beetle (Leptinotarsa decemlineata Say). We tested the effects of GBH and insecticide on the survival, insecticide target genes expression (acetylcholinesterase genes) and oxidative status biomarkers (glutathione S-transferase [GST], glucose-6-phosphate dehydrogenase [G6PDH], glutathione reductase homolog [GR], glutathione peroxidase homolog [GPx], total glutathione [totGSH], glutathione reduced-oxidized [GSH: GSSG], catalase [CAT], superoxide dismutase [SOD], lipid hydroperoxides). We found that exposure to indirect GBH has no single or interactive effects in combination with the insecticide on larval survival. However, prior exposure to GBH inhibits Ldace1 gene expression by 0.55-fold, which is the target site for the organophosphate and carbamate insecticides. This difference disappears when individuals are exposed to both GBH and insecticide, suggesting an antagonistic effect. On the other hand, oxidative status biomarker scores (PCAs of GPx, GR, and CAT) were decreased when exposed to both stressors, indicating a synergistic effect. Overall, we found that indirect GBH exposure can have both antagonistic and synergistic effects in combination with an insecticide, which should be considered when aiming for an ecologically relevant risk assessment of multiple human-induced stressors.

Western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), has developed resistance to transgenic corn that produces the insecticidal toxin Cry3Bb1 derived from the bacterium Bacillus thuringiensis (Bacillales: Bacillaceae) (Bt), with cross-resistance extending to corn with Bt toxins mCry3A and eCry3.1Ab. Additionally, some populations of western corn rootworm have evolved resistance to Cry34/35Ab1 corn. We conducted a 2-yr field and laboratory study that included three field locations: 1) Bt-susceptible population, 2) field with a recent history of Cry3Bb1 resistance, and 3) field with a long-term history of Cry3Bb1 resistance. The population with recently evolved Cry3Bb1 resistance showed resistance to Cry3Bb1 corn in both laboratory bioassays and field evaluations; by contrast, the population with a long-term history of Cry3Bb1 resistance showed resistance, in both laboratory and field experiments to Cry3Bb1 corn and corn with a pyramid of mCry3A plus eCry3.1Ab corn. Field-based evaluations also showed that the field population with a long-term history of Cry3Bb1 resistance imposed higher root injury to Cry3Bb1 corn and the pyramid of mCry3A plus eCry3.1Ab compared with the susceptible control. The results of this study are discussed in the context of developing strategies to manage western corn rootworm in areas where populations have evolved resistance to Cry3Bb1 corn.

Striacosta albicosta (Smith) (Lepidoptera: Noctuidae) is an important pest of corn, Zea mays L. in the Great Lakes region, which can be controlled by transgenic corn expressing Vip3A protein from Bacillus thuringiensis. To inform insect resistance management, the susceptibility, survival, and development of first, third, and fifth instar S. albicosta to Vip3A was determined using protein-overlay and corn tissue bioassays. Tissue bioassays were also used to determine the quantity of corn tissues with and without Vip3A-expression consumed by various instars. In diet bioassays, third and fifth instars were significantly less susceptible to Vip3A compared with first instars; however, no significant difference was observed in susceptibility of older instars. In tissue bioassays, survival was lowest for larvae fed Vip3A-expressing tissues, ranging from 0 to 21%, however, developmental measures of larvae fed Vip3A-expressing tissues did not differ from those fed artificial diet or tissues of other Bt events. Consumption of Vip3A × Cry1Ab tissues did not differ from that of Cry1Ab for each instar. Estimated Vip3A exposure of first instars ranged from 3 to 57 times higher than the concentration required for 99% mortality (LC₉₉) based on the product of the reported Vip3A expression in transgenic corn tissues and the consumption observed in tissue bioassays; however, the estimated exposure of third and fifth instars to Vip3A was lower than their respective LC₉₉. These findings suggest that first instar S. albicosta maybe exposed to a high dose of Vip3A under field conditions; however, Vip3A-expression in corn may not be high dose against older instars, increasing the risk of resistance development.

The common bed bug Cimex lectularius L. has undergone a worldwide expansion in recent years, due to increased opportunities for dispersal and development of insecticide resistance. For successful control, efficacy testing of products against bed bugs and determination of insecticide resistance under practical conditions are of outstanding importance. A new test system mimicking the practical use situation of residual insecticides was developed and evaluated. Bed bugs were attracted by CO₂ and heat to cross surfaces treated with alpha-cypermethrin and bendiocarb. In contrast to the complete efficacy of alpha-cypermethrin (less than 1% surviving bed bugs [with one exception of 5%]), only 45.3, 46, and 29% of insecticide-susceptible bed bugs showed lethal damage 7 d after contact with freshly bendiocarb-treated wallpaper or insecticide aged for 1 or 2 wk. Results show that the efficacy of different insecticides can be assessed with this new test system. Moreover, susceptibility to deltamethrin of five bed bug field strains, collected from infested apartments in Berlin, Germany, was determined in a filter paper contact bioassay. Resistance ratios (RRs) ranged between 4.3 and 20.7. In the novel simulated-use test, efficacy of alpha-cypermethrin was tested against the bed bug strain with the highest RR. In contrast to the insecticide-susceptible laboratory strain, alpha-cypermethrin was not effective against the field strain, where 26–50% of the bed bugs survived and even laid eggs. These results provide evidence for the presence of practically relevant pyrethroid resistance in bed bugs in Germany.

Metaflumizone is a novel semicarbazone insecticide. It functions as a sodium channel blocker insecticide (SCBI) with excellent insecticidal activity on most economically important lepidopterous pests. This study assessed the resistance risk of Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) to metaflumizone in the laboratory and the effects of metaflumizone selection on toxicities to other insecticides. Spodoptera exigua collected from a field population at Huizhou in 2012 were successively challenged by metaflumizone to evaluate the risk of resistance evolution.Twelve generations of selection increased resistance to metaflumizone by 3.4-fold and threshold trait analysis revealed that the realized heritability (h²) of this resistance was 0.086. When h² was equal to 0.086 and 90% of individuals were killed at each generation, LC₅₀ to metaflumizone increased by 10-fold after 15 generations. The selection by metaflumizone did not increase the resistance to indoxacarb, chlorantraniliprole, spinosad, methomyl, or endosulfan, suggesting a lack of cross-resistance. However, metaflumizone challenge upheld the recession of resistance to emamectin benzoate, chlorfluazuron, and tebufenozide.The block of resistance drops by metaflumizone exposure implied a possible cross-resistance between metaflumizone and these three insecticides. These results contribute to integrated resistance management of S. exigua.

Deciphering genetic structure and inferring migration routes of insects with high migratory ability have been challenging, due to weak genetic differentiation and limited resolution offered by traditional genotyping methods. Here, we tested the ability of double digest restriction-site associated DNA sequencing (ddRADseq)-based single nucleotide polymorphisms (SNPs) in revealing the population structure relative to 13 microsatellite markers by using four small brown planthopper populations as subjects. Using ddRADseq, we identified 230,000 RAD loci and 5,535 SNP sites, which were present in at least 80% of individuals across the four populations with a minimum sequencing depth of 10. Our results show that this large SNP panel is more powerful than traditional microsatellite markers in revealing fine-scale population structure among the small brown planthopper populations. In contrast to the mixed population structure suggested by microsatellites, discriminant analysis of principal components (DAPC) of the SNP dataset clearly separated the individuals into four geographic populations. Our results also suggest the DAPC analysis is more powerful than the principal component analysis (PCA) in resolving population genetic structure of high migratory taxa, probably due to the advantages of DAPC in using more genetic variation and the discriminant analysis function. Together, these results point to ddRADseq being a promising approach for population genetic and migration studies of small brown planthopper.

Methoprene-tolerant (Met) is a putative JH intracellular receptor that transduces JH signal by activation of the inducible Krüppel homolog 1 (Kr-h1). We analyzed the gene sequences of Met and Kr-h1 and their patterns of expression in Grapholita molesta (Busck) immature and adult stages in order to better understand the roles of these primary JH responders in regulating the metamorphosis and reproduction of this global pest of fruit crops. The deduced amino acid sequences of both GmMet and GmKr-h1 were highly homologous to those of other Lepidoptera, especially the cotton bollworm, Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae). Peak expression of GmMet occurred during the last 3 to 5 d of the final instar, followed by that of GmKr-h1, in the last 3 d of final instar. Similar patterns of GmMet and GmKr-h1 expression were detected across various tissue types in the fifth-instar larvae, with the highest expression observed in the head, followed by the epidermis, and the fat body. When expression of GmMet and GmKr-h1 was knocked down via dsRNA injection in the fifth instar, the results were increased larval mortality, abnormal pupation, delayed pupal duration, reduced adult emergence, extended preoviposition period, and reduced fecundity. We infer that both GmMet and GmKr-h1 participated in regulation of metamorphosis and reproduction in G. molesta, the former acting upstream of the latter, and could present biorational targets for novel pest control compounds.

Superoxide dismutase (SOD) is an antioxidant metalloenzyme that catalyzes the dismutation of the superoxide anion O₂^- to O₂ and H₂O₂. Many studies have focused on the role of SOD in response to abiotic stress, but its role during biotic stress, such as changes in organismal population density, has rarely been investigated. The oriental armyworm, Mythimna separata, is an economically important pest that exhibits phenotypic changes in response to population density. Solitary and gregarious phases occur at low and high population density, respectively. To examine the role of SODs in response to population density stress, we cloned two genes encoding SOD, MsCuZnSOD and MsMnSOD, and compared their expression in solitary and gregarious phases of M. separata. The MsCuZnSOD and MsMnSOD ORFs were 480 and 651 bp and encoded predicted protein products of 159 and 216 amino acids, respectively. The two SODs contained motifs that are typical of orthologous proteins. Real-time PCR indicated that the two SOD genes were expressed throughout developmental stages and were significantly upregulated in more mature stages of gregarious M. separata. Expression of the two SOD genes in various tissues of sixth-instar larvae was higher in gregarious versus solitary insects. Furthermore, expression of the SOD genes was significantly upregulated in response to crowding in solitary individuals, but suppressed in gregarious insects subjected to isolation. Collectively, these results suggest that population density may be key factor in the induction of SOD genes in M. separata.

African citrus greening disease (ACGD) is considered as one of the major diseases of citrus threatening citrus production in East Africa. Our study aimed for the first time to assess the incidence, severity, and distribution patterns of ACGD in Kenya and Tanzania. In total, 105 citrus orchards were assessed in 13 regions representing low, mid, and high altitude areas. In each backyard and orchard, trees were randomly selected and rated for visual ACGD symptoms; then leaves and insect samples collected for analysis of ‘Candidatus Liberibacter africanus' (CLaf), the presumptive causal agent of ACGD. Endpoint PCR, sequencing, and molecular phylogenetic tools were employed to confirm the identity of potential circulating pathogens. Incidence and severity of ACGD varied significantly among the different regions. Both Trioza erytreae (Del Guerico) (Hemiptera: Triozidae) and the invasive Asian citrus psyllid vector Diaphorina citri (Kuwayama) (Hemiptera: Liviidae) were found to co-occur in upper and lower midland regions. Molecular characterization identified ‘Candidatus Liberibacter africanus spp. Clausenae’ (CLafCl) as the main causal agent of ACGD in most of the citrus plants and insect samples. No instances of Candidatus Liberibacter asiaticus infection were found. These findings provide valuable insights into understanding and management of ACGD by employing stringent and early disease detection tools to curb the spread of the disease.

The agricultural landscape of North Dakota is changing. Corn and soybean are now commonplace, but once were rare. Spring sown wheat Triticum aestivum L. and durum wheat Triticum turgidum spp. durum continue to be dominant, but more winter-sown wheat is expected in the future. The presence of wheat in the landscape throughout much of the year will benefit populations of the Hessian fly, Mayetiola destructor (Say), which occurs throughout the state, sometimes in large numbers. Hessian fly is unusual among crop pests for which resources for plant resistance are well developed. On wheat genotypes expressing a single effective H resistance gene, 100% of larvae die before exhibiting any growth. Over 35 H genes in the public domain are available for crossing into elite cultivars. We explored the effectiveness of 31 Hessian fly resistance genes for a North Dakota Hessian fly population. Six genes—H4, H15, H21, H23, H26, and H29—caused 100% larval mortality. Seven others caused at least 80% mortality. Experimental data were used to address three additional questions. Do adult females avoid laying eggs on plants that will kill their offspring: Are neonate larvae able to detect resistance that will end up killing them? Do all 31 genes confer equal protection against larval-induced growth deficits? North Dakota wheat breeders have the necessary tools to create highly resistant wheat cultivars. So far, H genes have been deployed singly in cultivars. Advances in plant breeding will enable gene stacking, a more durable strategy over time.

Soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), infestations of soybean, Glycine max (L.) Merr. (Fabales: Fabaceae), and the associated yield loss have led to a large dependence on insecticidal management in soybean throughout the Midwestern United States. However, several populations of pyrethroid-resistant soybean aphids have recently been found in Iowa, Minnesota, North Dakota and South Dakota, which highlights the importance of alternative management approaches. One such alternative method is host-plant resistance, which uses naturally occurring plant defenses in crop cultivars to reduce the potential for yield loss from a pest population. Current soybean aphid-resistant cultivars do not protect against all soybean aphids due to the presence of virulent biotypes. In particular, soybean aphid biotype 4 is virulent to Rag1 and Rag2 resistance genes both individually and in combination. However, we hypothesized that resistance to biotype 4 may exist in previously identified, but uncharacterized resistant soybean plant introductions (PIs). To test this, we evaluated 51 previously identified but uncharacterized soybean aphid-resistant PIs for their resistance to colonies of soybean aphid biotype 4 collected in separate site-years (Lomira, WI 2013; Volga, SD 2015, 2016). Free-choice tests identified 14 PIs with putative resistance to ‘Lomira13′, two to ‘Volga15′, and eight to ‘Volga16′ soybean aphid colonies. Follow-up, no-choice tests corroborated two to three resistant PIs per colony, and PI 437696, which was resistant to each of the three colonies and could aid in breeding efforts and an integrated approach to soybean aphid management.

Cowpea, Vigna unguiculata (L.) Walp., serves as a major source of dietary protein in many tropical and subtropical regions around the world. To identify loci associated with agronomically desirable traits, eight elite cowpea cultivars were systematically inter-crossed for eight generations to yield 305 recombinant inbred lines. Here, we investigated whether these founder parents also possess resistance to the seed beetle Callosobruchus maculatus (F.), a highly destructive post-harvest pest. We estimated larval survival in seeds, egg-to-adult development time, adult mass at emergence, and seed acceptance for oviposition. Survival varied significantly among cowpea cultivars, but the pattern was complicated by an unexpected source of mortality; on three cultivars, mature larvae in a substantial fraction of seeds (20–36%) exited seeds prematurely, and consequently failed to molt into viable adults. Even if such seeds were eliminated from the analysis, survival in the remaining seeds varied from 49 to 92% across the eight parents. Development time and body mass also differed among hosts, with particularly slow larval development on three closely related cultivars. Egg-laying females readily accepted all cultivars except one with a moderately rugose seed coat. Overall, suitability ranks of the eight cultivars depended on beetle trait; a cultivar that received the most eggs (IT82E-18) also conferred low survival. However, one cultivar (IT93K-503-1) was a relatively poor host for all traits. Given the magnitude of variation among parental cultivars, future assays of genotyped recombinant progeny can identify genomic regions and candidate genes associated with resistance to seed beetles.

Members of the highly polyphagous Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) species complex cause major crop damage by feeding and by transmitting plant viruses.The Middle East-Asia Minor 1 (MEAM1) of the B. tabaci complex is by far the most problematic whitefly affecting crops including cultivated watermelon (Citrullus lanatus; Cucurbitaceae: Cucurbitales). Watermelon cultivars share a narrow genetic base and are highly susceptible to whiteflies. We studied the potential of C. ecirrhosus, a perennial desert species that can be hybridized with C. lanatus, as a source of whitefly resistance for cultivated watermelon. The results of this study indicate that C. ecirrhosus offers resistance (although not total) against the MEAM1 B. tabaci based on, at least, antibiosis and antixenosis. Whitefly performance concerning developmental survival, body size attainment, and nonpreference were suppressed on C. ecirrhosus compared with the watermelon cultivar ‘Sugar Baby’. Also, our olfactometer results indicated that the adults were less attracted to leaf volatiles of C. ecirrhosus. Although there is a pungent odor associated with the leaves of C. ecirrhosus, the leaf volatiles had no toxic effect on adult whitefly survival as compared with cultivated watermelon. We also demonstrated that plants of C. ecirrhosus can be clonally propagated from vine cuttings of the parent plant. Using traditional breeding procedures, C. ecirrhosus was hybridized with C. lanatus and viable F₁ and F₂ seeds were produced. This is the first report of pest resistance in C. ecirrhosus. This wild species offers a source of resistance against whiteflies for the improvement of cultivated watermelon.

The small tomato borer, Neoleucinodes elegantalis (Guenée), is an important pest of tomato (Solanum lycopersicum L.) in South and Central America. This pest is a potential threat for many tomato-producing areas and was listed in 2014 as an A1 quarantine pest by the European and Mediterranean Plant Protection Organization. Soon after hatching, the neonate N. elegantalis larvae penetrate the fruits where they feed until pupation.Therefore, effective N. elegantalis management relies on the timely scouting of egg densities to allow decision-making prior to penetration of neonates into fruits. This study aimed to develop a conventional sampling plan for scouting N. elegantalis eggs on tomato plants. The most representative and precise sampling unit was the most basal fruit cluster for plants bearing up to three clusters (S1 plants). For plants with more than three fruit clusters (S2 plants), the most representative and precise sampling unit was the combination of the second and third fruit clusters. Among the four variables evaluated (eggs/fruit, egg masses/fruit, percentage fruit with eggs, and percentage clusters with eggs), the percentage of clusters with eggs was the most economical for N. elegantalis sampling based on the number of samples and cost required. For this variable, the number of samples determined at the 25% error level was 42 and 36 samples for S1 and S2 plants, respectively. The sampling plan developed for scouting N. elegantalis is fast, reflects pest infestation in tomato fields, and costs less than US$1.50 per field scouted.

Laboratory tests on acclimated and nonacclimated life stages of Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae) (adults, pupae, larvae, and eggs) and Oryzaephilus surinamensis (L.) (Coleoptera: Silvanidae) (adults, larvae, and eggs) were conducted at 0, –5, –10, and –15°C to evaluate effects of acclimation on susceptibility to cold treatment. Acclimation of all tested life stages for 7 d at 15°C affected susceptibility of both species to the cold temperatures. After 1 d exposures for ≥2 h, acclimated adults had a noticeable increase in cold tolerance compared with nonacclimated adults for both tested species. Nonacclimated pupae of T. confusum were equally susceptible to cold compared with acclimated pupae at short exposures to low temperatures. Exposure of nonacclimated life stages of T. confusum, at –10°C for 1 d gave 0% survival. Similarly, almost all (99.6%) nonacclimated individuals of O. surinamensis died at –10°C. At 0°C, nonacclimated larvae were more cold tolerant than acclimated larvae, but this trend was reversed when larvae were exposed to –5°C. Mixed results were obtained for larvae of O. surinamensis because in some of the combinations tested, nonacclimated larvae were more tolerant, even at temperatures that were lower than 0°C. In contrast to O. surinamensis, eggs of T. confusum that had not been exposed to cold were not affected by acclimation, while exposure to cold showed increased cold hardiness in acclimated eggs. Results show that individual stored-product insect species may have mixed susceptibility to cold temperatures, which must be taken into account when using cold treatment as a management strategy.

We investigated the efficacy of two types of modified atmospheres (MA) against adults of the khapra beetle, Trogoderma granarium Everts, and the maize weevil, Sitophilus zeamais Motschulsky, under laboratory conditions. Adults of the above species on wheat were exposed to a carbon dioxide (CO₂) concentration of 70% or a low oxygen (O₂) concentration of 0.1% for durations of 0.67 (16 h), 1, 2, 4, and 6 d and stored in an environmental chamber set at 28 ± 2°C, 70 ± 5% RH, and a photoperiod of 16:8 (L:D) h. After each exposure interval, immediate mortality and knockdown were recorded, and the surviving or knocked down individuals were transferred to normal atmospheres and returned to the environmental chamber, where survival was recorded 7 d later. In addition, after the immediate and delayed mortality counts, all adults were removed from the substrate, and the number of progeny produced was recorded 60 d later. Both MA conditions totally controlled (100% mortality) the adults of T. granarium and S. zeamais immediately after a 6-d exposure or after a 4-d exposure when delayed mortality was taken into account, showing the postexposure effect of the MA. Moreover, high CO₂ was more effective than low O₂ for S. zeamais, whereas the reverse was true for T. granarium. The 4-d exposure period was crucial for the production of progeny of both species because after that period surviving insects did not produce offspring. Our results show that both MA conditions can be used with success to control these species.

Historically, stored product insect pest management has been based around the use of methyl bromide and phosphine as fumigants. However, methyl bromide has been phased out for structural fumigations, and there is increasing worldwide insecticide resistance to phosphine. One potential alternative, environmentally friendly option is the use of methyl benzoate (MBe), which is considered a food safe compound. In this study, we evaluated the direct and sublethal effects of MBe exposure on the survivorship and mobility of four stored product species with diverse life histories, including Rhyzopertha dominica, Tribolium castaneum, Sitophilus zeamais, and Trogoderma variabile. Sets of insects were exposed to a control, low, or high MBe concentrations in flasks with or without food for 24 or 72 h in the laboratory. Furthermore, we assessed phosphine exposure under similar conditions. Overall, R. dominica was the most susceptible to MBe exposure, followed by T. castaneum. By contrast, S. zeamais and T. variabile were relatively unaffected by MBe exposure. Exposure to MBe induced multiple-fold decreases in the total distance moved and velocity of adults still considered alive or affected after assays. By comparison, phosphine effectively killed all individuals of all species. Our data suggest that while MBe was effective for R. dominica, it was not competitive in comparison to phosphine for controlling susceptible strains of these species at the specified experimental conditions.

The stable fly, Stomoxys calcitrans (L.), is a bloodsucking ectoparasite that causes irritation and distress to livestock, wildlife, and humans. Both sexes are vicious blood-feeders that feed on a variety of animals. Optically attractive sticky traps have been used to capture stable flies, and some companies claim that sticky traps can protect animals from the bites of stable flies. To further investigate the protective ability of sticky traps, Home and Garden Mosquito (HGM) traps were selected for evaluations at the University of Florida Horse Teaching Unit (HTU). Broodmares coated with fluorescent dust were either tethered to a post in the center of a paddock or released untethered into a paddock. HGM sticky traps were placed at the four compass points and four selected distances from the paddock center to capture stable flies before (unmarked) or after (marked) they visited the horses. More than 40% of flies captured on traps placed closest to the horses were marked. This indicates that the traps did not prevent the flies from visiting the horses. A percentage of marked and unmarked stable flies showed signs of blood in their guts indicating recent feeding. For unknown reasons, the number of stable flies marked with Signal Green dust exceeded the numbers marked with other colors. Although the HGM traps caught ample numbers of stable flies, the traps did not prevent stable flies from feeding on the horses. More work is needed to determine optimal trap placement and densities required to maximize stable fly management with traps.

The mortality rate of a field population of house fly (Musca domestica L.) was determined for a granular fly bait containing the active ingredient indoxacarb, which was compared to two commercially available granular fly baits containing either dinotefuran or cyantraniliprole. Indoxacarb was applied at three different application rates 0.498, 0.986, and 1.972 g/m² (low, medium, and high). Time to 50% mortality was fastest for dinotefuran (5.7 h) and slowest for the low application rate of indoxacarb (10.3 h). Time to 90% mortality was fastest for the high application rate of indoxacarb (27.7 h) and slowest for dinotefuran (51.0 h) and cyantraniliprole (45.9 h). Among the three indoxacarb application rates, the high rate reached both 50 and 90% fly mortality significantly faster than the low rate. The medium rate did not significantly differ from either the high or low application rates. Dinotefuran bait produced greater fly mortality than all other treatments at 30-min post-exposure, with mortality for remaining baits exceeding controls by 3- to 6-h post-exposure. All insecticidal baits produced similar fly mortality by 6-h post-exposure and >94% fly mortality by 96-h post-exposure, indicating that each may be effective in a fly management program. Flies consumed a similar amount of the indoxacarb (regardless of application rate) and dinotefuran baits, but consumed less of the cyantraniliprole bait, suggesting a feeding irritancy or toxicity effect manifested during consumption. Nevertheless, flies consumed enough cyantraniliprole bait to cause mortality similar to other baits by 6-h post-exposure.

Pests and disease have become an increasingly common issue as globalized trade brings non-native species into unfamiliar systems. Emerald ash borer (Agrilus planipennis), is an Asiatic species of boring beetle currently devastating the native population of ash (Fraxinus) trees in the northern forests of the United States, with 85 million trees having already succumbed across much of the Midwest. We have developed a reaction-diffusion partial differential equation model to predict the spread of emerald ash borer over a heterogeneous 2-D landscape, with the initial ash tree distribution given by data from the Forest Inventory and Analysis. As expected, the model predictions show that emerald ash borer consumes ash which causes the local ash population to decline, while emerald ash borer spreads outward to other areas. Once the local ash population begins to decline emerald ash borer also declines due to the loss of available habitat. Our model's strength lies with its focus on the county scale and its linkage between emerald ash borer population growth and ash density. This enables one to make accurate predictions regarding emerald ash borer spread which allows one to consider various methods of control as well as to accurately study the economic effects of emerald ash borer spread.

The newly developed fungal biopesticide Aprehend, containing spores of Beauveria bassiana, is the first biological control agent to be incorporated into management programs to control the common bed bug (Cimex lectularius L.) (Hemiptera: Cimicidae). Aprehend is sprayed as barriers where bed bugs are likely to walk and pick up spores as they search for a bloodmeal. A key application target for Aprehend is the box spring, which may be covered by encasement-type or insecticide-impregnated covers. Since some insecticides can reduce the persistence of fungal spores, we tested the efficacy and spore germination percentages of Aprehend when applied to the two types of box spring covers. We found that spore germination was about 11% lower on the permethrin-impregnated ActiveGuard cover than on the encasement-type AllerEase cover. However, bed bugs exposed for 15 min to Aprehend on the two box spring covers suffered similarly high levels of mortality irrespective of the cover material. Thus, there was no inhibitory or additive effect of the ActiveGuard cover on bed bug mortality. Lastly, overall mortality was higher if bed bugs were exposed to Aprehend-treated ActiveGuard than the ActiveGuard cover alone. Our findings indicate that if pest managers are using ActiveGuard covers in combination with Aprehend, best practice would be to use ActiveGuard on mattresses and apply Aprehend directly to the box spring or to a box spring covered by an encasement-type cover.

Formulations of entomopathogenic (insect-killing) fungi represent alternatives to synthetic insecticides in the management of forest and shade tree insects. We evaluated bark spray applications of the entomopathogen Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Cordycipitaceae) strain GHA (BotaniGardES), permethrin insecticide (Astro), and water (control) on colonization of black walnut (Juglans nigra L.) (Fagales: Juglandaceae) bolts by the walnut twig beetle (Pityophthorus juglandis Blackman) (Coleoptera: Curculionidae), vector of the fungus that causes thousand cankers disease. Treated bolts were baited with a P. juglandis aggregation pheromone lure and deployed in infested walnut trees. Bark application of permethrin prevented P. juglandis colonization of the phloem. Although treatment of bolts with the B. bassiana suspension did not reduce P. juglandis colonization or short-term emergence relative to the control treatment, it increased the B. bassiana infection rate from 25 to 62% of emerged adults. Results suggest that commercial applications of B. bassiana strain GHA may help augment natural levels of infection by this entomopathogen in the eastern United States, and support continued exploration of entomopathogens for biological control of the walnut twig beetle.

Techniques for the survey of necrophagous dipterans are frequently designed for adult flies and often neglect forensically important information, such as the postfeeding larval dispersal following carcass colonization. In this study, we propose and test a novel trap design for the collection of dipteran larvae at the postfeeding stage. The Trap for Dispersing Larvae (TDL) consists of two semiindependent parts: 1) a 2-liter PET bottle, containing the attractant (carcass); and 2) a plastic box, which selectively captures postfeeding dispersing larvae. Based on a field test in a dry forest fragment in Brazil, the TDL trap captured ca. 1,500 larvae of 13 species of Calliphoridae, Muscidae, Fanniidae, and Sarcophagidae. The species richness of dispersing larvae recorded in our study was ca. 60% of that previously recorded in this area. As the TDL trap can assess larval dispersal without interfering with carcass decomposition, we demonstrate that it can be used to assess temporal variation in the dynamics of colonization. In the present study, most larval dispersal occurred at 5-d postexposure, but the traps remained efficient until 7 d, by which time rat carcasses had skeletonized. We conclude that the TDL trap captures the dispersing dipteran larvae with little interference in carcass decomposition processes. The trap also provides a representative sample of species and is simple to use and of low cost.

Both the parasitoid wasp Spalangia endius Walker and the insecticide imidacloprid are used to control house flies. A recent study found that negative sublethal effects of imidacloprid on killing flies and on offspring production by this parasitoid wasp are eliminated when females have the opportunity to crawl through decaying matter. An enzyme-linked immunosorbent assay showed that the presence of decaying matter reduces the amount of pesticide on their bodies. This study examined whether this was also true for sublethal effects on mating. S. endius were exposed to a realistic concentration of imidacloprid that induces very low mortality. Then, individual parasitoids were allowed to burrow through decaying organic matter or not, followed by mating tests in the absence of decaying matter. Even after 24 h with the decaying matter, copulation for both males and females that had previously been exposed to imidacloprid was delayed compared with no-pesticide controls. Furthermore, for pesticide-exposed males, subsequently burrowing through media made copulation even more delayed than if they were not exposed to media. For pesticide-exposed females, subsequently burrowing through media neither increased or decreased the negative effect of the pesticide exposure. Together with other studies, these results reinforce that use of S. endius and use of imidacloprid are incompatible, even at much lower than recommended concentration, unless application is sufficiently separated in place and time.

Brown wheat mites, Petrobia latens (Müller 1776, Acari: Tetranychidae), are sporadic yet economically damaging pests of winter cereals. In Colorado, their life history is closely tied to the development of winter wheat, where they are present in the field from crop planting in late September through harvest in early June. In order to withstand winter months, these mites are able to survive cold temperatures. However, the mechanisms of cold hardening and their temperature limits are unknown. This research documents the seasonal supercooling points of the brown wheat mite. Their seasonal average supercooling point stayed consistent throughout the year, never varying more than a degree from the overall average supercooling point of -17°C. The greatest variation in supercooling point was seen in the spring, during which supercooling point temperatures ranged from -9.2 to -25.5°C. We also documented the upper and lower lethal temperatures for the brown wheat mite. When comparing small nymphs to large nymph and adult stages, small nymphs were slightly more cold tolerant (lethal temperature estimates required to kill 99% of the population [LT₉₉] were -30.8 and -30.6°C, respectively), but less heat tolerant (LT₉₉ was 50 and 56°C, respectively).