Journal of Economic Entomology

Environmental RNAi (eRNAi) is a recent innovation in insect pest control, and comprehensive risk assessment is needed to ensure the environmental safety and longevity of this technology. As eRNAi relies on the insects cellular machinery for its mode of action, environmentally mediated plasticity in the activity of cellular processes required for RNAi could influence efficacy and the development of resistance. Here, we investigated the extent to which plant cultivar and temperature influence the efficacy of insecticidal double-stranded RNA (dsRNA) targeting actin in larvae of the Colorado potato beetle (CPB; Leptinotarsa decemlineata). Potato cultivar did not significantly affect survival or gene silencing in dsRNA-treated larvae, indicating that efficacy is consistent across potato varieties, at least under laboratory conditions. However, the amount of leaf tissue consumed by larvae varied across cultivars, with consumption being inversely proportional to protein content. Thus, the ingested dose of dsRNA may vary across crop varieties. Temperature did influence RNAi efficacy, with both gene silencing and mortality being reduced when dsRNA treatment occurred at lower temperatures. After three days of feeding with dsRNA, gene silencing occurred at all temperatures, but knockdown efficiency was 62% at 30{degrees}C and 35% at 18{degrees}C. Beetles consumed significantly less leaf tissue (and thus dsRNA) at 18{degrees}C, but the observed temperature-dependent effects could not be fully explained by the quantity of dsRNA ingested. Further, efficacy at different temperatures was not related to transcript levels of core RNAi genes, indicating that other mechanisms are responsible for the observed effects. Overall, these results indicate that environmental conditions can influence the efficacy of insecticidal eRNAi and may affect the rate at which insects develop resistance to these technologies.

Given that plant nutrient content is both spatially and temporally dynamic (Lenhart et al., 2015; Deans et al., 2016, 2018), insect herbivores are exposed to an incredible amount of nutritional variability. This variability can constrain insects to feeding on sub-optimal resources, but it can also provide an opportunity for insects to regulate their intake of specific nutrients to obtain an optimal balance. Nutrient regulation has implications for pest control strategies in agricultural systems, as the nutritional state of pest species may impact their susceptibility to insecticides. Deans et al. (2017) showed that diet macronutrient balance has significant effects on the susceptibility of Helicoverpa zea larvae to Cry1Ac, an endotoxin expressed in transgenic Bt crops. This was demonstrated using a highly inbred laboratory strain of H. zea, limiting the applicability of these results to field populations that encompass greater genetic diversity. In this study, we assessed the impact of field-relevant macronutrient variability on the efficacy of two Bt endotoxins, Cry1Ab and Cry1Ac, using three field populations collected from different geographic regions. This was done to further understand the impact of nutritional variability on Bt susceptibility and also to determine the relevance of these effects in the field. While we saw limited differences in Cry susceptibility across populations, dietary effects were highly variable. Across populations there were distinct population-level differences in the interactions between Cry concentration and diet, the type of Cry toxin impacted by diet, and the treatment diet that produced optimal survival and performance. These results show that nutrition can have strong impacts on Bt susceptibility but also that these impacts are strongly affected by genetic background in H. zea. To accurately assess Bt susceptibility in the field, including resistance monitoring, bioassay methods should incorporate the appropriate nutritional parameters and be as localized as possible.

O_LIHerbivorous insects are major crop pests whose feeding often results in significant economic damage. Detecting and monitoring insect herbivory is crucial for effective pest management, but manual methods can be time-consuming, inefficient, and destructive. This study investigates the use of contact microphones as a non-invasive and efficient tool for detecting, identifying, and monitoring insects feeding on plants. C_LIO_LIApproach and methods: Contact microphones connected to a Raspberry Pi were clipped onto plant stems and programmed to record sounds made by the feeding insects. Three combinations of herbivorous insects and plants were evaluated: tobacco hornworm (Manduca sexta Linnaeus) on tobacco, Colorado potato beetle (Leptinotarsa decemlineata Say) on potato, and European corn borer (Ostrinia nubilalis Hubner) and Northern corn rootworm (Diabrotica barberi Smith & Lawrence) on corn (Zea mays Linnaeus). Sound recordings were analyzed to determine the presence and absence of insects and to quantify differences in feeding activity between different insects. The laboratory study was repeated in a corn field to observe corn rootworms (Diabrotica spp.). C_LIO_LIMain results: Contact microphones successfully detected insect herbivory on the tobacco, potato, and corn plants in the lab and field, demonstrating the use of substrate-born acoustic signals for detecting both large and small-bodied insect pests. The recordings also revealed differences in the feeding patterns, frequency, and amplitude between insect species. Field experiments indicate low background noise relative to lab experiments. C_LIO_LIConclusions and implications: Contact microphones offer a promising cost-effective, non-destructive, and efficient method for monitoring insect herbivory. This approach has the potential to improve scientific observations of insect herbivores and pest management strategies by enabling early pest detection and measuring the success of interventions. Early field results indicate the viability of this approach under realistic conditions in the field. The results highlight the importance of considering the acoustic environment in agricultural ecosystems, suggesting sound can be used to better understand the behavior of insects and their interactions with plant hosts. Contact microphones could have widespread applications in agriculture and ecology by providing a simple and effective tool for detecting and monitoring insect activity. C_LI

Meloidogyne enterolobii is an aggressive root-knot nematode (RKN) species that has emerged as a significant pathogen of sweetpotato in the Southeastern United States. M. enterolobii is spread primarily through the movement of infected seed sweetpotatoes used for propagation. The RKN resistance in commercially grown sweetpotato cultivars has proven ineffective against this nematode. Detecting RKN in sweetpotato by eye is unreliable, and further distinguishing M. enterolobii from other RKN species that infect sweetpotato is labor intensive; relying on molecular tests conducted on individual nematodes dissected out of host roots by trained technicians. Here, we have developed a high-throughput survey method to collect skin samples and extract total DNA from batches of sweetpotato storage roots. Combining this method with species-specific PCR assays allowed for quick and sensitive detection of M. enterolobii and other RKN species infecting sweetpotatoes. We tested this method using batches of infected storage roots at varying levels of M. enterolobii infection. We also inoculated skin samples with varying numbers of individual M. enterolobii eggs to determine the methods detection threshold and used this method to conduct surveys for RKN on fresh market sweetpotatoes. Our results show that this method can consistently and reliably detect M. enterolobii in sweetpotato batches at levels as low as 2 eggs per 10 mL skin sample. This method will be a useful tool to help screen for the presence of M. enterolobii in seed sweetpotatoes before they are replanted, thereby helping to slow the spread of this nematode to M. enterolobii-free sweetpotato growing operations.

The spotted wing drosophila (SWD) Drosophila suzukii (Diptera: Drosophilidae) is a pest of soft fruit. Since its introduction in Europe in 2008 farmers struggle to protect their crops. The sterile insect technique (SIT) has proven efficient at controlling numerous fruit fly species and could be deployed to control D. suzukii. In recent years, key elements of SIT applied to D. suzukii have become available. However, field- and field-like experiments are scarce. In this experiment, we assayed the efficacy of a high-performance strain at reducing the reproduction of D. suzukii in complex, yet replicated and controlled conditions. Two ratios of sterile to fertile insects (5:1 and 1:1) using bisexual releases were compared to a control treatment with fertile, wild flies only. The presence of sterile individuals at a 5:1 ratio significantly reduced fly reproduction, measured after 5 days, by an approximate threefold factor. However, the proportion of infested fruits in the treated plots remained unaffected. The number of available berries in the cage appeared an unexpected determinant of fly infestation, suggesting undocumented density-dependent processes. The success of this assay opens the door to larger scales experiments, over several generations, and, in the near future, the field-evaluation of the efficacy of the SIT to control D. suzukii.

The alfalfa leafcutting bee (Megachile rotundata) is one of the primary pollinators for the alfalfa seed industry. The alfalfa leafcutting bee is a solitary cavity nesting bee. Female Megachile rotundata bees will construct and provision individual brood cells lined with cut leaves (cocoon) and will gather nectar and pollen to place within the constructed cocoon. The female bee will lay a single egg within the constructed cocoon and leave the egg to undergo larval stage development and pupation into the adult stage. During this time multiple pathogens and parasitoids can prey on the developing larvae, resulting in the loss of the future adult bee. A major concern for commercial alfalfa seed growers is the presence of invertebrate pests and fugal pathogens. In the present study, we used historical data from the Parma Cocoon Diagnostic Laboratory to determine baseline rates of pathogen and parasite infection of Megachile rotundata cells and used this analysis to determine historical infection rates and cutoffs for management practices. Additionally, using a Faxitron (X-ray) analysis for Megachile rotundata cell obtained in 2020, we compared the presence of chalkbrood, pathogens, and parasitoids in samples collected from both growers stocks and newly purchased Canada bees. The results of the investigation demonstrate historical averages of the presence of chalkbrood, pathogens, and parasitoids. We also show a significant increase in chalkbrood and predators in 2007-2011 and a significant difference in chalkbrood and predators between bee samples obtained from Canada and grower stocks.

The global application of Transeius montdorensis (Acari: Phytoseiidae) as a biological control agent across various protected crops has proven effective against a range of insect pests like thrips and whiteflies, as well as pest mites like broad mites and russet mites. Optimization of rearing T. montdorensis under laboratory conditions is crucial for further studies of this species to improve their application in Integrated Pest Management (IPM) programs. Here, we evaluated the development and reproduction of T. montdorensis when fed on four different diets, including cattail pollen (Typha latifolia), living dried fruit mites (Carpoglyphus lactis), frozen C. lactis eggs, and a mixed diet of frozen C. lactis eggs and T. latifolia pollen. Females consuming the mixed diet exhibited superior total fecundity and daily oviposition rate, along with the highest intrinsic rate of increase (rm) and net productive rate (R0) among all diets tested. The immature period was significantly longer for mites on a diet of living C. lactis compared to those on other diets. Importantly, utilizing frozen C. lactis eggs and T. latifolia pollen mitigates the risk of infestation and contamination from the living dried fruit mites, which is important for laboratory and field settings when releasing the predator colonies. Our findings not only present an optimized rearing method for predatory mites under laboratory conditions but also suggest potential broader applications for enhancing the effectiveness and sustainability of biological control strategies across various agroecosystems and reducing dependency on chemical pesticides.

Dalbulus maidis [(DeLong & Wolcott), corn leafhopper], a phloem-feeding insect, is the most efficient vector of maize stunting pathogens (Spiroplasma kunkelii, Maize bushy stunt phytoplasma, and Maize rayado fino virus) in the Americas. Studies involving gene editing in insects are rapidly providing information that can potentially be used for insect vector and plant disease control. RNA interference (RNAi), a sequence-specific gene silencing method, is one of the most widely used molecular tools in functional genomics studies. RNAi uses exogenous double-stranded RNA (dsRNA) or small interfering RNA (siRNA) to prevent the production of proteins by inhibiting the expression of their corresponding messenger RNA (mRNA). In this study, we measured the efficacy of gene silencing, and its effects on D. maidis mortality as proof of concept that RNAi is a viable tool for use in genetic pest control of phloem-feeding insects. Oral delivery of dsRNA using an artificial diet was used to silence two key insect genes, vacuolar ATP synthase subunit B, and subunit D (V-ATPase B and V-ATPase D). Our results showed reduced gene expression of V-ATPase B and V-ATPase D after ingestion of dsRNA, and significantly higher mortality, and wing deformation, associated with reduced gene expression, compared to control insects that were not orally fed dsRNA. These results reveal RNAi as a viable tool for use in genetic pest control of phloem-feeding insects, and a way for further functional genomic studies, such as identification of potential target genes for either population suppression or population replacement of this vector of maize diseases.

O_LIBees are important for vital pollination of wild and crop plants, but are in decline worldwide. Intensification of agriculture is a major driver of bee decline. Organic farming practices are designed to limit environmental impacts of agriculture and can increase bee abundance and species diversity. However, studies have been heavily focused towards some guilds of bees, overlooking others. This includes social brood parasites, cuckoo bumblebees, an understudied bee lineage. Little is known about bumblebee host and cuckoo population dynamics, and the effects of farming practice on cuckoo bumblebees have never previously been evaluated.\nC_LIO_LITo compare the effects of farming practice (organic vs conventional) on the abundance, species diversity, and community dissimilarity of cuckoo bumblebees and their hosts, we compared host and cuckoo community metrics across ten matched pairs of organic and conventional farms in Yorkshire, UK.\nC_LIO_LIAs found by many previous studies, host bumblebees were more abundant on organic farms than on conventional farms. Despite this, cuckoo bumblebees were equally abundant on both farm types. Contrary to prediction, community dissimilarity and species diversity were unaffected by farm type for both host and cuckoo communities.\nC_LIO_LISynthesis and applications: Results suggest that cuckoo bumblebee community metrics are not solely driven by host community metrics, and that cuckoos may respond differently from their hosts to differences among farming practices. This could, in turn, indicate that a unified management practice is not sufficient to conserve all bumblebee species.\nC_LI

Honey bees contribute substantially to the world economy through pollination services and honey production. In the U.S. alone, honey bee pollination is estimated to contribute at least $11 billion annually, primarily through the pollination of specialty crops. However, beekeepers lose about half of their hives every season due to disease, insecticides, and other environmental factors. Here, we explore and validate a spatiotemporal statistical model of Varroa destructor mite burden (in mites/300 bees) in managed honey bee colonies, exploring the impact of both environmental factors and beekeeper behaviors. We examine risk factors for Varroa infestation using apiary inspection data collected across the state of Illinois over 2018-19, and we test the models using inspection data from 2020-21. After accounting for spatial and temporal trends, we find that environmental factors (e.g., floral quality, insecticide load) are not predictive of Varroa intensity, while several beekeeper behaviors (e.g., smaller colony density, supplemental feeding, and mite monitoring/treatment) are protective against Varroa. Interestingly, while monitoring and treating for Varroa is protective, treating without monitoring is no more effective than not treating at all. This is an important result supporting Integrated Pest Management (IPM) approaches. Author SummaryHoney bees contribute substantially to the world economy through pollination services and honey production. However, beekeepers lose about half of their hives every season due to disease, insecticides, and other environmental factors. Pathogens, such as Varroa mites and the viruses they vector, are especially detrimental to colony health, and best practices for pest management remain contentious. In this study, we model Varroa destructor mite burden in managed honey bee colonies using apiary inspection data collected across the state of Illinois from 2018 - 2021. Our modelling approach accounts for both spatial and temporal trends, allowing us to investigate the marginal impacts of environmental factors and beekeeper interventions on mite burden. We show that treating for Varroa mites has a protective effect only when accompanied by a monitoring strategy, important evidence in favor of Integrated Pest Management (IPM) approaches.

Myzus persicae, a serious sap-sucking pest of a large variety of host plants in agriculture, is traditionally controlled using chemical insecticides but there is interest in using biopesticides as restrictions are increasingly placed on the use of broad-spectrum pesticides. Here we show that in petri dish experiments high concentrations of the fungal entomopathogen Beauveria bassiana (strain PRRI 5336) lead to rapid mortality of M. persicae but at a low concentration (1 x 104 conidia mL-1) there is a hormetic effect where longevity and fecundity are enhanced. Hormetic effects persisted across a generation with reduced development times and increased fecundity in the offspring of M. persicae exposed to B. bassiana. Whole plant experiments point to a hormetic effect being detected in two out of three tested lines. The impact of these effects might also depend on whether M. persicae was transinfected with the endosymbiont Rickettsiella viridis, which decreases fecundity and survival compared to aphids lacking this endosymbiont. This fecundity cost was ameliorated in the generation following exposure to the entomopathogen. While B. bassiana is effective in controlling M. persicae especially at higher spore concentrations, utilization of this entomopathogen requires careful consideration of hormetic effects at lower spore concentrations, and further research to optimize its application for sustainable agriculture is recommended. AUTHOR SUMMARYBiopesticides such as Beauveria bassiana can be effective alternatives to chemical insecticides to control insect pests. We tested the efficacy of this biopesticide against the important agricultural pest aphid Myzus persicae in laboratory experiments. We also tested whether the potential biological control agent and endosymbiont Rickettsiella viridis could provide protection against mortality caused by B. bassiana. While high doses of B. bassiana caused rapid mortality in aphids, low doses enhanced aphid fecundity and survival. This enhancement persisted into the next generation, with shortened development times and increased fecundity regardless, even when high doses were used in the previous generation. The endosymbiont R. viridis did not provide clear protection against B. bassiana, in contrast to previous studies in other aphid species, but beneficial effects at low doses also occurred in this aphid line. We also observed hormesis on experiments on whole plants, but only for some aphid genotypes. To a lesser extent, we also observed beneficial effects of low doses of B. bassiana in experiments on whole plants, but only in some aphid genotypes. Fitness enhancement by biopesticides at low doses raises concerns for field applications but further research is required to understand its underlying mechanisms.

The most devastating pest to honey bees (Apis mellifera) worldwide is the parasitic mite Varroa destructor. The development of miticide-resistant mite populations has been a major driver of colony loss in many countries. We investigated the threat Varroa poses to honey bee populations in New Zealand and tested the effectiveness of the two most popular chemical treatments used by beekeepers. Colony losses reported by New Zealand beekeepers have risen over five consecutive years from 2017 to 2021, as have the proportion of losses attributed to Varroa, with this parasite found to be the main driver of colony loss in 2021. Varroa resistance to miticide treatments flumethrin and amitraz was tested. The concentration of flumethrin required to kill 50% of the mites (LC50) was 156 g/g, 13 times greater than the adjusted LC50 value of 12 g/g observed in a trial also conducted in New Zealand in 2003, thus indicating evidence of developing mite resistance to flumethrin in New Zealand. Molecular analyses searching for mutations in the Varroa genome known to be associated with flumethrin resistance found no evidence of such mutations, suggesting that any extant resistance to flumethrin has evolved independently in New Zealand. No evidence of resistance to amitraz was found, as the LC50 value of 12 g/g was lower than what was observed in the 2003 trial (110 g/g). Further development of integrated pest management, such as gene-silencing RNA interference (RNAi) and selective breeding of Varroa-resistant bees, is needed to effectively manage a parasite that threatens global agriculture.

Istocheta aldrichi (Diptera: Tachinidae) is a specialist parasitoid of the invasive Japanese beetle, Popillia japonica (Coleoptera: Scarabaeidae). Research and releases for biological control depend on field collecting parasitized hosts and rearing the parasitoid through diapause to obtain I. aldrichi adults. This study investigated how rearing practices before, during and after the seasonal overwintering period affected the proportion of I. aldrichi pupae that emerged as adults, the timing of parasitoid emergence, and their longevity. Increasing cold exposure duration during overwintering increased adult I. aldrichi emergence from puparia and reduced development time after transfer to warm conditions. Adult I. aldrichi emergence from overwintered puparia depended on interactions between overwintering environment (indoors vs. outdoors), spring thermal regime, and the timing of host collection in the previous season. Burying puparia in the soil in late summer/early fall resulted in higher subsequent adult I. aldrichi emergence. Manipulating spring temperatures in controlled environments allowed parasitoid emergence to be staggered over several weeks without reducing emergence success. Emergence under outdoor spring conditions was unreliable. Adult longevity was affected by temperature and diet: cooler conditions extended lifespan, honey-water increased longevity relative to pollen alone or honey-water and pollen together. These results provide a foundation to further improve I. aldrichi rearing techniques for use in experimental research and applied biological control of P. japonica.

The cucurbit beetle Diabrotica speciosa is one of the most important corn pests in Brazil, seen as responsible for about 10-13% losses in maze crops, especially in the southern region of Brazil. The development of sustainable technologies for specific pest control, such as RNAi, depends on the identification of target genes and on the characterization of the gene expression profile of these genes in different developmental stages. Gene expression analyses, on the other hand, depend on reliable reference genes for the normalization of RT-qPCR data. In this study, we performed a cross-species evaluation of qPCR reference genes already validated in Diabrotica virgifera virgifera, due to its relevance as an important corn pest in North America. The expression of the reference genes EF1, {beta}-actin, {beta}-tubulin, and GAPDH was evaluated in different developmental stages of the Diabrotica speciosa: egg, neonate, 3rd instar larva, pupa, and adult. The stability of the reference genes was analyzed using the {Delta}Ct method, BestKeeper, GeNorm, and NormFinder algorithms available at RefFinder web-based tool. From all four genes, {beta}-actin and EF1 were the most suitable for normalizing genes, with both expression and stability ranking position in accordance with the results reported for Diabrotica virgifera virgifera. Our findings provide valuable information for researchers studying gene expression during different developmental stages of this species and highlight the potential of validating cross-species reference genes.

BACKGROUNDThe implementation of sterile insect technique (SIT) has proven effective in the area-wide suppression of several significant agricultural and sanitary pests by employing traditional cobalt-60 (60 Co-{gamma}) as a radiation source. Recently, X-ray has been validated as a feasible alternative to 60 Co-{gamma} radiation sources. Nonetheless, higher doses of X-ray irradiation lead to insect sterility but diminish mating competitiveness, thereby impacting the effectiveness of SIT applications. Thus, it is crucial to ascertain the optimal irradiation dose and develop strategies to enhance the mating competitiveness of sterile insects to enhance SIT efficacy. RESULTSIn this study, we determined the effect of various X-ray irradiation doses (ranging from 0 to 366 Gy) on the fecundity, fertility, and mating competitiveness of Cydia pomonella, a globally invasive fruit pest. Results demonstrated that the sterility rate of sterile males increased proportionally with irradiation dose up to 200 Gy, beyond which it plateaued. Notably, exposure to 200 Gy of irradiation notably decreased the mating competitiveness of male, as evidenced by a mating competitiveness index of 0.17 in laboratory and 0.096 in the orchard. This decline in mating competitiveness is likely linked to the down-regulation of genes associated with the recognition of sex pheromones, specifically CpomOR3a, CpomOR3b, and CpomOR5, following X-ray irradiation. Fumigation of the plant volatile, linalool at varying concentrations (70, 83, and 96 L/m 3) resulted in differential enhancements in male mating competitiveness, with the moderate concentration significantly improving the competitiveness of sterilized males, possibly by restoring their ability to recognize sex pheromones. Implementation of repeated releases of sterilized males on a pilot scale led to a notable reduction in the population of C. pomonella in the field. CONCLUSIONThese findings indicate that fumigation with plant volatiles has the potential to mitigate male sterility induced by X-ray irradiation, offering a promising approach to enhance the efficacy of SIT applications for the control of C. pomonella. Graphic Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=110 SRC="FIGDIR/small/604520v2_ufig1.gif" ALT="Figure 1"> View larger version (27K): org.highwire.dtl.DTLVardef@7ac9aeorg.highwire.dtl.DTLVardef@14ed866org.highwire.dtl.DTLVardef@1611e8borg.highwire.dtl.DTLVardef@11c6377_HPS_FORMAT_FIGEXP M_FIG C_FIG We determined the optimal X-ray irradiation dose and developed a linalool fumigation approach to improve the mating competitiveness of sterilized insects, thereby bolstering the efficacy of against Cydia pomonella.

BACKGROUNDEstimating parasitoid abundance in the field can be difficult, even more so when attempting to quantify parasitism rates and the ecosystem service of biological control that parasitoids can provide. To understand how observed parasitism rates (in-field mummy counts) of the green peach aphid, Myzus persicae (Sulzer) (Homoptera: Aphididae) translate to actual parasitism rates (laboratory-reared parasitoid counts), field work was undertaken in Australian canola fields over a growing season. Parasitoids were reared within a controlled laboratory setting. RESULTSTotal observed and actual parasitism rates of M. persicae varied considerably across regions, but less so on a field level. Overall, actual parasitism was on average 2.4 times higher than that observed in the field, with rates an average of 4-fold higher in South Australia. As crop growth stage progressed, the percentage of mummies observed increased. Percentage of parasitoids reared also increased with crop growth stage, averaging 3.4% during flowering and reaching 14.4% during podding/senescing. Although there was a greater diversity of reared parasitoid species at later crop growth stages, actual parasitism rate was unaffected by parasitoid species. Diaeretiella rapae was the most commonly reared parasitoid, increasing in abundance with crop growth stage. CONCLUSIONThese findings indicate that mummy counts alone do not provide a clear representation of parasitism within fields.

Aphid parasitoids (Hymenoptera; Aphidiidae) were surveyed within grain production landscapes in Victoria, Australia between 2017 and 2018, as well as more sporadically nationwide between 2016 and 2019. In addition, aphidiine records were collated from insect depositories around Australia and online databases. The 5551 specimens recorded constituted a total of 23 species and seven genera. Diaeretiella rapae (MIntosh) was the most common species, representing more than 70% of all aphidiines recorded. This species also showed a greater northerly geographic range than other aphidiines. During sampling between 2017 and 2019, aphidiines were reared from mummies to ascertain host-parasitoid relationships. Diaeretiella rapae was again the most commonly reared parasitoid, although aphidiine preference varied with aphid host and between states and territories. An illustrated dichotomous key to Australian aphidiines in grain production landscapes is provided for the 11 species sampled in our field surveys. This is the first comprehensive review of aphidiines sampled within Australia in over two decades. Knowledge about the diversity and distribution of these parasitoids is important for understanding their impact on current and future invasions of aphid species. In addition, understanding the interactions between grain aphids and their associated parasitoids will further support the inclusion of parasitoid wasps into integrated pest management (IPM) strategies.

Trichogramma, a genus of egg parasitoid wasps, are widely used as biological control agents and serve as model organisms in parasitoid research. Despite their significance, the understanding of RNA interference (RNAi) in Trichogramma remains very limited. In this study, we investigated RNAi-associated genes by bioinformatic approaches and experimentally assessed the feasibility of RNAi and the susceptibility of environmental RNAi in Trichogramma. We found that Trichogramma genomes contain a complete set of genes in the RNAi pathway and exhibit extensive gene expansion of dsRNase, which may influence RNAi efficiency by degrading dsRNA. We demonstrated successful RNAi through pupal microinjection in T. dendrolimi Matsumura, providing a technical approach for future gene functional studies. In addition, we observed no evidence of susceptibility to environmental RNAi in either T. dendrolimi adults or larvae, which might be attributed to the extensive expansion of dsRNase. This low environmental RNAi sensitivity in Trichogramma could suggest a reduced risk of RNAi-based pest management strategies affecting nontarget Trichogramma populations. Overall, this study presents a technical approach for conducting gene functional studies in Trichogramma and provides a foundation for evaluating the nontarget effects of RNAi-based pest control strategies on Trichogramma.

Tetranychus urticae (Koch) is an economically important pest of many agricultural commodities in the Pacific Northwest. Multiple miticides are currently registered for control including abamectin, bifenazate, bifenthrin, and extoxazole. However, populations of Tetranychus urticae have developed miticide resistance through multiple mechanisms, in many different growing regions. Producers of agricultural commodities where Tetranychus urticae infestations are problematic rely on integrated pest management tools to determine optimal control methods. Within this species multiple single nucleotide polymorphisms have been documented in different genes which are associated with miticide resistance phenotypes. The detection of these mutations through TaqMan qPCR has been suggested as a practical, quick, and reliable tool to inform agricultural producers of miticide resistance phenotypes present within their fields and have potential utility for making appropriate miticide application and integrated pest management decisions. Within this investigation we examined the use of a TaqMan qPCR-based approach to determine miticide resistance genotypes in field-collected populations of Tetranychus urticae from mint fields and hop yards in the Pacific Northwest of the United States and confirmed the results with a multiplex targeted sequencing. The results suggest the TaqMan approach accurately genotypes Tetranychus urticae populations collected from agricultural fields. The interpretation of the results, however, provide additional challenges for integrated pest management practitioners, including making miticide application recommendations where populations of Tetranychus urticae are a mix of resistant and wildtype individuals.

A critical component of integrated pest management is minimizing disruption of biological control by reducing use of pesticides with significant non-target effects on natural enemies. Insecticide non-target effects testing for natural enemies has become increasingly common, but research examining the non-target effects of herbicides on natural enemies is scarce and recommendations regarding herbicide selectivity are non-existent. We used meta-analysis to summarize laboratory bioassays testing non-target effects of herbicides on arthropod natural enemies and identify patterns in taxon susceptibility and active ingredient toxicity. Data was extracted from 103 papers representing 801 total observations. Herbicides increased natural enemy mortality and decreased longevity, reproduction, and predation. Mesostigmatan mites and hemipterans were the most sensitive to herbicides and spiders, neuropterans, and hymenopterans were the least sensitive. Mortality was higher in juvenile predators versus parasitoids, but did not differ between adults; parasitoid juveniles are likely better protected within the host. In terms of acute mortality, metribuzin, glufosinate, and oxyfluorfen were the most harmful herbicides. Only nicosulfuron, rimsulfuron, pendimethalin, phenmedipham, atrazine, and urea did not increase natural enemy mortality. The large effect size of glufosinate is particularly concerning, as it is the most likely replacement herbicide for glyphosate in many crops. Many active ingredients remain under-studied. Our analysis indicates that herbicides have a strong potential to disrupt biological control in cropping systems. Simple SummaryReducing the use of pesticides that harm natural enemies of crop pests is important to pest management. Currently, there is limited information on how herbicides might affect natural enemies. The researchers found that herbicides increased natural enemy mortality and reduced their longevity and efficacy as predators. Some potential glyphosate replacement herbicides were more harmful than glyphosate. There was little or no data available for many herbicides and beneficial insects, indicating that much more research is needed on this topic.