Integrating entomopathogenic fungi (EPF) into crop protection programs requires ensuring their compatibility with insecticides, herbicides, and fungicides to enable simultaneous applications and reduce operational costs. Compatibility is typically assessed by incorporating pesticides into growth media and measuring toxicity through germination, growth, and sporulation (Biological Index). However, field applications mainly expose conidia directly to pesticides, potentially leading to mismatches between laboratory and real-world outcomes. Here, we evaluated the compatibility of 5 fungicides (copper oxychloride, fosetyl + proamocarb, potassium phosphonate, difenoconazole, and fluopyram + tebuconazole) and 4 herbicides (diflufenican, oxyfluorfen, MCPA, and glyphosate) with 2 virulent EPF strains (Metarhizium brunneum and Beauveria bassiana) selected for efficacy against olive fruit fly, Bactrocera oleae (Rossi) (Diptera: Tephritidae), comparing the conventional medium-amendment approach (MAA) with a direct conidial-pesticide mixture method (CPM), and relating both to in vivo biological activity. For M. brunneum, only one of 4 herbicides was compatible using MAA, whereas CPM classified all 4 as compatible. Of the 5 fungicides, MAA classified 3 as compatible and 2 as toxic, while CPM classified 3 as compatible and 2 as moderately toxic. In vivo bioassays using adults of the tephritid model Mediterranean fruit fly, Ceratitis capitata (Wiedemann) indicated that CPM results aligned more closely with observed efficacy. B. bassiana showed similar results, with lower virulence and fungicide compatibility, and CPM again better reflecting in vivo biological activity. Overall, results suggest that CPM may provide a more realistic approach than MAA for assessing EPF compatibility with agrochemicals and could facilitate EPF integration into crop protection.

Grapevine red blotch virus (GRBV) reduces grapevine vigor, delays grape ripening, reduces sugar content and quality of wines in North America and other continents. To date, the three-cornered alfalfa hopper, Spissistilus festinus, has been clearly identified as a vector in the United States. Since this hopper has not been observed in some vineyards where secondary spread occurs, additional vectors of GRBV are likely. To search for vectors, we conducted a two-year survey collecting live hemipterans from infected vines and surrounding grass, trees, and shrubs. Screening a wide variety of hemipterans from a vineyard with infected plants focuses on relevant insects and avoids the difficulty of rearing multiple species. Most collected insects were immediately used in greenhouse transmission tests to infect clean grapevines, and a subset of insects were frozen for PCR tests. Out of 147 transmission tests using 245 plants, four plants were infected after exposure to the leafhoppers Euscelidius variegatus or Dikraneura absenta or delphacid Nothodelphax sp. Whole-body PCR tests of both E. variegatus and D. absenta frozen directly from the field were positive confirming that they ingested GRBV from the field (no Nothodelphax tested). PCR tests of the cixiid Cixius sp. and leafhoppers Fieberiella florii and Gyponana octolineata were positive but the grapevine transmission tests did not support these as vectors. This survey has identified three hemipterans for further transmission studies and for specific GRBV tests of salivary glands to fully confirm vectoring potential.

Sugars are essential nutrients for the growth, development, and reproduction of insects. To enhance the reproductive and pest control potential of Trichogramma pintoi, this study investigated the effects of supplementing different concentrations (10%, 15%, 20%, and 25%) of nutrients, sucrose, glucose, fructose, maltose, trehalose, honey, and water (control), on the longevity, fecundity, flight capacity, and gustatory response of T. pintoi adults. The results showed that T. pintoi took the shortest time to locate 20% concentrations of the different nutrients and exhibited the longest feeding duration. Supplementing fructose (female 19.53 d, male 11 d) and sucrose (female 17.67 d, male 9 d) resulted in the longest adult longevity of T. pintoi, with the highest fecundity (131.73 eggs/female, and 137.4 eggs/female, respectively). While, T. pintoi fed on water had a shortest longevity (Female: 1.47 d, Male: 1.27 d), and a lowest fecundity (12.93 eggs/female). Additionally, supplementing with different sugar solutions significantly enhanced the flight capability of T. pintoi. Among them, honey produced the highest proportion of L4 (flight capability with Level 4), reaching 78.67%. This study demonstrated that supplementing with 20% fructose and sucrose can significantly prolong the longevity and fecundity of T. pintoi, while honey can significantly enhance their flight capacity. These findings provide a reference standard for improving the mass rearing capacity and application effcacy of T. pintoi pest control.

The hedgehog grain aphid (HGA), Sipha maydis Passerini (Hemiptera: Aphididae), is a significant pest of cereal crops and is present in the United States. Like other aphids, it exhibits wing polymorphism, producing both winged and apterous morphs influenced by environmental factors such as temperature and host plant quality. This study evaluated the effects of temperature and host plant species on life history traits and wing induction under controlled laboratory conditions. Two weeks after nymphs being placed on host plants, a significant interaction between host plant and temperature was observed for number of aphids and winged morph proportion. The highest aphid populations occurred on wheat and barley at 30 °C (356 ± 70 and 344 ± 42, respectively), while sorghum supported consistently lower populations, peaking at 15 °C (37 ± 4). Wing induction was greatest on sorghum and barley at 25 °C (26.9 ± 7.5% and 10.1 ± 1.6%, respectively), while no differences were observed among temperatures on wheat (x̄ = 9.2 ± 2.0%), suggesting that the host may have induced a stress-related morphological response to promote dispersal, particularly on sorghum, the least suitable host. Supercooling points (SCPs) of winged and apterous adults reared at 25 °C were also measured. While SCPs did not differ significantly between morphs, they varied by host plant: aphids reared on wheat froze at -16.7 °C, compared to -18.7 °C on sorghum and -18.6 °C on barley, indicating host-mediated cold tolerance. These findings underscore the ecological plasticity of HGA and provide valuable insights for improving outbreak prediction and integrated pest management strategies across diverse environmental conditions.

Titanium dioxide nanoparticles (TiO2 NPs) are widely used in industrial, agricultural, and consumer products and are increasingly detected in terrestrial and floral environments that are critical for pollinators. However, the mechanistic and ecological consequences of TiO2 NPs exposure on pollinator health remain poorly understood. Here, we present a multi-level ecotoxicological assessment of dietary TiO2 NPs exposure in the bumblebee Bombus terrestris L. (Hymenoptera: Apidae), integrating colony performance, gut microbiota composition, and host transcriptomic responses. Using both environmentally relevant and worst-case concentrations, we conducted short-term (3-d) and chronic (15-d) exposures to evaluate physiological disruption, microbial dynamics, and potential compensatory or recovery processes. TiO2 NPs ingestion reduced worker survival and syrup consumption, delayed oviposition, and shortened egg development, indicating impaired colony performance. Gut microbiota exhibited pronounced dysbiosis after 3 d, followed by partial converged toward control-like profiles after 15 d, suggesting microbial potential resilience and compensatory restructuring. Transcriptomic analyses revealed dose-dependent activation of immune, oxidative stress, calcium signaling, and programmed cell death pathways. Correlative analyses identified associations between specific microbial taxa, including Apibacter and Klebsiella, and host gene expression linked to immunity, detoxification, and metabolism, highlighting coordinated microbe-host responses under TiO2 NPs stress. This study provides novel mechanistic insight into how engineered nanoparticles affect pollinators across biological scales, from colony physiology and microbial ecology to molecular pathways, and emphasizes the practical importance of safeguarding pollinator health for both wild plant reproduction and crop pollination. The findings advance understanding of nanoparticle ecotoxicology and support evidence-based strategies for sustainable nanomaterial use and pollinator protection.

Spiders are major predatory natural enemies in agricultural and forest ecosystems and play an important role in pest control. The vitellogenin receptor (VgR) mediates the endocytosis of vitellogenin (Vg) into developing oocytes, providing essential nutrients for embryogenesis and playing a critical role in insect reproductive processes. In this study, A VgR gene was cloned from adult female Pardosa astrigera (L. Koch) and designated as PastVgR1. The open reading frame of PastVgR1 is 5,337 bp in length and encodes a protein of 1,779 amino acids, with a predicted molecular weight of 197.8 kDa. Sequence analysis revealed that the PastVgR1 protein possesses an N-terminal signal peptide consisting of the first 17 amino acid residues and contains conserved domains characteristic of the LDLR superfamily. RT-qPCR analysis showed that PastVgR1 expression was highly specific to the ovaries, with the highest levels in virgin females. RNA interference (RNAi)-mediated silencing of PastVgR1 significantly prolonged the pre-oviposition period, caused ovarian atrophy and oocyte dispersion, reduced both the number of eggs laid per female and the hatching rate, and delayed ovarian development. In summary, PastVgR1 represents a promising molecular target for enhancing the mass rearing of predatory spiders used in biological pest management. Optimizing the large-scale production of these natural enemies could lower the cost of biological control and ultimately support the development of sustainable, green agriculture.

Fungal secondary metabolites and virulence factors are central to the ecological success of entomopathogenic fungi (EPF) and represent untapped resources for sustainable pest management, especially against sucking insect pests. EPF initiates infection by penetrating the insect cuticle, then deploys a repertoire of toxic metabolites into the hemocoel to suppress host defenses and accelerate mortality. Key examples, such as beauvericin from Beauveria bassiana and destruxin from Metarhizium anisopliae, exemplify how these compounds function as both potent virulence factors and promising biocontrol agents. Structurally diverse metabolites-including nonribosomal peptides, alkaloids, terpenes, and polyketides-are synthesized via tightly regulated biosynthetic pathways that optimize pathogenicity. For decades, the genetic and biochemical mechanisms governing these processes remained largely unexplored. However, breakthroughs in next-generation sequencing and targeted genetic engineering have unveiled critical genes and regulatory networks underlying metabolite production and host-fungus interactions. This knowledge is enabling the rational enhancement of EPF strains and the strategic deployment of their metabolites in pest control programs. Harnessing fungal metabolites with optimized virulence offers a targeted, environmentally safe alternative to conventional insecticides, with reduced risks to nontarget organisms and minimized chemical residues. Continued research into the molecular ecology, biosynthetic innovation, and field performance of these compounds will be pivotal in translating laboratory discoveries into robust, field-ready biocontrol solutions for managing economically important insect pests.

Certain invasive ambrosia beetles are attracted to and colonize physiologically stressed trees emitting ethanol. Xylosandrus germanus (Blandford) and Xylosandrus crassiusculus (Motschulsky) are attracted to trees emitting ethanol and are 2 of the most damaging invasive ambrosia beetles in North America. Acetic acid has been detected in stressed trees emitting ethanol, but whether it has a role in ambrosia beetle ecology is unknown. Sections of tree stems (bolts) were soaked in various dilutions of acetic acid alone and mixed with a 5% dilution of ethanol, then deployed in a field near a woodland. Bolts soaked in 5% or 10% dilutions of acetic acid were more attractive to X. germanus and X. crassiusculus than bolts soaked in dilutions of 0.1% or 1.0%. Dilutions of acetic acid mixed with 5% ethanol were more attractive to X. germanus and X. crassiusculus than similar dilutions of acetic acid alone or 5% ethanol alone. Furthermore, more offspring were produced in bolts soaked in mixtures of acetic acid and 5% ethanol than either compound alone. This information helps improve our knowledge of the physiological conditions in trees that make them suitable for colonization by X. germanus and X. crassiusculus. Furthermore, the increased attraction of damaging ambrosia beetles to acetic acid + ethanol could be useful for improving baits used for monitoring emergence of X. germanus and X. crassiusculus in spring.

Bumblebees are efficient pollinators of fruits and vegetables in greenhouses and field crops. However, pesticide use in agricultural landscapes is causing a sharp decline in pollinating insect populations. The impact of pesticides on bumblebee health is a growing concern. Cyfluthrin, atrazine, and prothioconazole are 3 commonly used pesticides in agricultural production. Although the Food and Agriculture Organization of the United Nations has published the acute median lethal dose (LD50) data for these 3 pesticides on Apis mellifera honey bee, there is still a lack of LD50 data for non-Apis bees, such as Bombus terrestris (Linnaeus, 1758). Therefore, this study determined the oral median lethal dose (LD50) of 3 pesticides, cyfluthrin, atrazine, and prothioconazole, in European bumblebees (Bombus terrestris). The active ingredient of each pesticide was first dissolved in dimethyl sulfoxide and then diluted in a sucrose solution to prepare the pesticide-sucrose mixture for feeding. The oral LD50 values of cyfluthrin for worker bees were 4.27, 3.36, and 2.16 μg/bee at 24, 48, and 72 h, respectively. The 24-h LD50 for virgin queens was 13.49 μg/bee. For 24-h exposures in worker bees, the oral LD50 values of atrazine, prothioconazole, and their mixture were 355.3, 530.0, and 480.4 μg/bee, respectively. Pesticide-sucrose solution intake decreased as pesticide concentration increased. This study provides a preliminary evaluation of the toxicity of 3 pesticide types on bumblebees and offers insight for improving the conservation and sustainability of pollinators in agriculture. Additionally, the findings contribute to regulatory assessments by providing crucial data on pesticide effects on B. terrestris, supporting more comprehensive and effective pesticide regulations.

The bird cherry-oat aphid, Rhopalosiphum padi (Linnaeus; Hemiptera: Aphididae), is a major cereal pest and a vector within the barley yellow dwarf virus complex. Manual scouting is labor-intensive and can be inconsistent, motivating rapid, image-based monitoring. We developed R. padi Count, an optimized deep learning framework built on Ultralytics YOLO11 for automated detection and image-level counting of R. padi in cluttered imagery. Starting from a YOLO11n baseline, we integrated an ADown downsampling module, Triplet Attention, and a Focusing Diffusion Pyramid Network module to improve small-object discrimination while limiting computational overhead. The model was trained and evaluated on smartphone images acquired under semi-controlled greenhouse conditions that captured variable backgrounds and occlusion. Relative to evaluated YOLO baselines, R. padi Count improved detection accuracy and reduced counting error, achieving a mean average precision at an IoU threshold of 0.50 (mAP50) of 92.97%, with an image-level mean absolute error of 1.86 aphids per image and a root mean squared error of 3.50 aphids per image. These results support the feasibility of practical, image-based aphid monitoring in the studied setting and provide a foundation for future evaluations of cross-scenario robustness and on-device performance in deployment contexts.