Edible insects are increasingly recognized as sustainable and nutrient-dense ingredients with potential applications across diverse food systems. While their use in solid foods has been widely explored, the incorporation of insect-derived ingredients into beverages remains fragmented and insufficiently conceptualized. This narrative review critically examines the current state of insect-based beverages, integrating technological, nutritional, cultural, and market-oriented perspectives. Rather than adopting a fully systematic review methodology, this article synthesizes representative scientific literature, traditional practices, and emerging commercial examples to explore how edible insects are being positioned within two distinct yet complementary pathways: (i) functional beverages targeting nutrition, gut health, sports performance, immunity, and meal replacement, and (ii) experiential beverages driven by culinary storytelling, tradition, ritual, and sensory innovation, such as insect-infused beers and spirits. Relevant sources were identified through searches of major scientific databases using combinations of keywords such as edible insects, entomophagy, insect-based beverages, functional beverages, and insect protein, with emphasis on peer-reviewed literature published mainly over the past two decades, complemented by representative examples of traditional practices and emerging commercial products. The review discusses the nutritional and biofunctional potential of insect-derived proteins, lipids, and chitin-related components, with particular emphasis on their behavior in liquid matrices, including solubility, stability, flavor impact, and processing constraints. Technological challenges associated with beverage formulations are critically assessed. In parallel, consumer perception is examined through the lens of neophobia, sensory expectations, and the role of cultural framing and gastronomy in facilitating acceptance. By bridging food science, beverage technology, and food culture, this review identifies key knowledge gaps and research priorities for advancing insect-based beverages beyond niche applications. It argues that future development in this field will depend not only on technological optimization and a regulatory framework, but also on the deliberate design of products that integrate functionality with meaningful culinary and cultural narratives.
The global expansion of tree plantations has led to extensive fragmentation of natural forests, posing significant challenges for biodiversity conservation. Understanding the diversity patterns and underlying mechanisms of ground-dwelling insects in these fragmented landscapes is critical to inform effective conservation strategies. To address this, we sampled ground-dwelling insects using pitfall traps across nine remnant natural forest fragments ("islands") embedded within a tree plantation matrix in Guangxi, China. We examined insect family-level diversity and community composition in relation to fragment isolation (low vs. high) and size (large vs. small) and explored the mechanisms driving the observed patterns. Our results revealed no significant difference in ground-dwelling insect diversity between low-isolation and high-isolation fragments. However, diversity was significantly lower in smaller fragments compared to larger ones. This reduction was primarily driven by decreased seedling density within smaller fragments, directly reflecting the adverse effects of plantation-driven fragmentation on native seedling establishment. Furthermore, we observed noble shifts in community composition of ground-dwelling insects along both fragment isolation and size gradients. Highly isolated fragments exhibited a decline in phytophagous insects and omnivores (with detritivore-herbivore diets), but an increase in detritivores. Smaller fragments exhibited consistent declines across multiple insect taxa spanning various dietary guilds. The observed changes in ground-dwelling insect composition were driven by shifts in plant (especially seedling) community composition. Our findings reveal a clear cascading effect: plantation-driven fragmentation limits native plant regeneration, and these limitations subsequently propagate to higher trophic levels, profoundly impacting ground-dwelling insects. Effective restoration of plantation-fragmented landscapes requires strategies that both prioritize the preservation of large, continuous forest fragments and promote native seedling recruitment within existing fragments.
The rapid expansion of edible insect production has focused primarily on rearing, processing efficiency, safety, and nutritional composition, while the slaughter of insects has received comparatively little scientific and ethical scrutiny. This narrative review examines insect slaughter as a critical control point linking bioethics, physiology, and ingredient quality. The review synthesizes evidence from neurobiology, food science, and processing studies to evaluate how commonly used slaughter methods interact with biological aspects of insects. Existing literature shows that slaughter techniques influence protein stability and hydrolysis, lipid oxidation, antioxidant retention, techno-functional properties such as emulsification and gelation, as well as sensory attributes and consumer acceptance. Available evidence suggests that methods designed to rapidly suppress metabolic activity may be associated with improved preservation of certain nutritional and functional parameters, although findings remain species- and context-dependent. The review further highlights major knowledge gaps, including the lack of species- and life-stage-specific welfare indicators and standardized assessment protocols. Overall, the findings support the need to reconceptualize insect slaughter as a strategic upstream decision rather than a neutral processing step. Integrating ethical considerations with nutritional, functional, and regulatory perspectives is essential for establishing science-based standards and ensuring the responsible development of edible insect-based food and feed systems.
Insect sex pheromone receptors (PRs) are crucial for regulating mating and reproduction. In the insect olfactory perception pathway, the pheromone-binding protein (PBP) facilitates the efficient translocation of sex pheromones, enabling them to bind to PRs. PRs convert chemical signals into electrical signals, which are transmitted to the insect central nervous system to ultimately regulate reproductive behaviors. Thus, conducting functional analysis of PRs not only clarifies the molecular mechanism underlying insect mating via sex pheromone recognition and reveals the intrinsic regulatory link between sex pheromone detection and mating behavior but also provides theoretical support for the scientific understanding of the insect olfactory system. Additionally, this research lays a core theoretical foundation for the development of green pest control technologies in agriculture and forestry. This paper systematically reviews the research methods, technical principles, and advantages and disadvantages of techniques used to study insect PR genes. It summarizes representative identified PRs and their corresponding research strategies, aiming to provide a reference for future investigations into insect chemical communication and for the advancement of pest control practices.
Insecticides in various formulations are indispensable for agricultural pest control and the prevention of vector-borne diseases. Piperonyl butoxide (PBO) is a synergist widely used to enhance the effectiveness of insecticides, notably pyrethroids, by inhibiting the detoxifying cytochrome P450 enzymes, thus reducing the capacity of insects to metabolize and resist insecticides. Recent studies have revealed an unexpectedly weak enhancement of pyrethroid efficacy by PBO, which may have serious implications for insect control. The underlying mechanism of this effect remains unclear. Here, we demonstrate that the PBO-induced inhibition of cytochrome P450 impedes the effect of deltamethrin by mainly affecting its interaction with the inactivated state of voltage-gated sodium channels (Nav). We describe a new octopamine-dependent regulatory mechanism involving Gαs, PKA, DARPP-32, and PP1-2A, which affect the cytochrome P450 conformation, thus limiting the effect of PBO and modulating the Nav gating. As a result, deltamethrin cannot reach its final binding site in the fenestration to exert its full effect. We confirmed in vivo that the octopamine level is elevated in insects under the treatment with a chemical stressor, which decreases the deltamethrin efficacy. Our findings reveal a novel adaptive mechanism that increases insect survival by reducing insecticide efficacy, underscoring the need to develop more effective formulations and technologies for pest and vector control.
Cytochrome P450 enzymes (P450s) play a key role in the metabolism of a diverse array of exogenous compounds. In insects they are often implicated in the phase I metabolism of many insecticides, which can lead to the emergence of resistance. Studies of such P450s have frequently employed heterologous expression in insect cell lines for in vitro functional characterisation. However, identifying optimal conditions for insect P450s expression is challenging. Here, we address this challenge by developing an improved protocol for insect P450s expression in insect cell lines employing two new concepts: CPR Balance and P450 Point. By adjusting the viral titer of CPR/P450, stable expression of microsomal P450 in the cellular system was achieved; By establishing time gradients for viral infection of H5 cells, the optimal time point for P450 harvest was determined. These two improvements enhanced the operability of P450 in vitro expression and increased the content of active P450. Using three P450s from agricultural pest silverleaf whitefly, Bemisia tabaci, we demonstrated that the protocol is efficient and stable in expressing active P450 proteins in vitro and is applicable to multiple CYPs from different subfamilies. Additionally, a detailed step-by-step protocol is included in the Appendix. Our work provides an improved reference for eukaryotic expression of P450s from insects and will inform future in vitro expression work using the bac-to-bac system.
Rapidly expanding insect farming industries worldwide are raising concerns about introducing contaminants into food chains. One of the dominant species in this new bioindustry, the black soldier fly (Hermetia illucens), offers diverse benefits ranging from protein production to waste management, biofuel, and pharmaceutical applications. Recent studies have highlighted the ability of insects to accumulate metals; however, knowledge of their ability to eliminate contaminants is needed. Similarly, information regarding the accumulation of polycyclic aromatic hydrocarbons (PAHs) is lacking. This study aimed to understand how H. illucens larvae accumulate and eliminate metals and PAHs from contaminated substrates. We performed two-phase bioaccumulation experiments followed by toxicokinetic modelling to estimate uptake and elimination rate constants and half-lives in H. illucens. During the uptake phase, insects were exposed for five days to contaminated substrates at EU maximum feed concentrations: 2 mg kg-1 for As and Cd, 10 mg kg-1 for Pb, and 12.5 μg kg-1 for each of four PAHs (benzo[a]pyrene, benz[a]anthracene, benzo[b]fluoranthene, chrysene). In the second phase, insects were exposed to non-contaminated substrates for five days to evaluate elimination. Results showed metal accumulation with kinetic bioaccumulation factors of 4.26, 1.16, and 1.31 for Cd, As, and Pb, respectively. A one-day depuration reduced As and Pb below regulatory thresholds; Cd required three days. Minimal to no accumulation was observed for B[a]P and B[b]F, with higher uptake for B[a]A. PAH half-lives were under one day. These findings, from controlled experiments with limited contaminants, support incorporating a depuration phase for risk mitigation in H. illucens waste-to-protein systems.
The single von Willebrand factor C (SVWC) domain-containing protein family represents a crucial class of immune molecules recently identified in insects and crustaceans. Initially regarded as functional analogs of vertebrate interferons (IFNs) due to their virus-induced expression and activation of the Janus kinase-signal transducer and activator of the transcription (JAK-STAT) pathway, recent studies have revealed that SVWC proteins possess far more complex functions. Many SVWC members are themselves a novel class of pattern recognition receptors (PRRs) that can directly bind to viruses and bacteria. Importantly, SVWCs are not a single entity but a highly diverse family-multiple subtypes exist in Drosophila, Bombyx mori, and shrimp-a gene expansion that implies functional differentiation. This review systematically examines the multifunctionality of SVWC proteins in insects and crustaceans, with a particular focus on the functional specialization driven by subtype diversity. We delve into the complex regulatory networks governing SVWC expression, including the differential activation by nuclear factor kappa B (NF-κB) pathways (Dorsal, Rel-2, Relish) and interferon regulatory factor (IRF) pathways. We detail the unique signaling mechanism by which SVWCs activate the JAK-STAT pathway via integrins, rather than the canonical Domeless receptor. Furthermore, we extend the discussion to the emerging roles of SVWCs as PRRs in humoral immunity (activating Toll/IMD pathways to induce antimicrobial peptides) and cellular immunity (mediating hemocyte phagocytosis). Based on current evidence, We propose that diverse SVWC subtypes may recognize distinct pathogens, bind to different integrin receptors, and activate specific STAT variants via disparate upstream induction pathways, thereby establishing a systematic and hierarchical immunoregulatory network. This understanding positions the SVWC protein family as a central hub in the insect immune network and offers a novel perspective on the complexity and evolution of invertebrate immunity.
A technology that enables highly sensitive, selective on-site detection of odorants without sophisticated equipment could be valuable for a range of applications, from water quality testing to disease diagnosis. In this study, we introduce a portable technology using Sf21 cells engineered to express Drosophila melanogaster odorant receptors, co-receptors, and calcium-sensitive fluorescent proteins for geosmin detection-a natural compound known for its distinct musty odor. Sensor cartridges were fabricated by immobilizing sensor cells on the glass surface inside tubes. When coupled with a portable fluorometer, the system successfully detected geosmin at concentrations as low as 100 pM (18.2 ppt) without requiring any concentration steps. Moreover, the system was deployed at a lakeside dam and detected geosmin in real field samples. This is the first successful detection of target odorants using biosensors comprising transfected sensor cells bearing specific odorant receptor genes. The results highlight the practicality of insect odorant receptor-expressing sensor cells and reveal the potential of this technology in various applications, such as food, beverage, and water quality testing, where on-site odorant detection is required.
Trait-environment relationships are a promising tool to understand how environmental factors shape freshwater communities, particularly on large scales. However, inconsistent responses of the community trait composition to environmental gradients have been reported for freshwater invertebrates. One reason for these discrepancies may be the predominant use of single traits without accounting for intercorrelations among traits. In this study, we evaluated whether grouping taxa into trait profile groups (TPGs), i.e., groups with similar trait combinations, yields more consistent relationships with estimated pesticide toxicity across large spatial scales compared to single traits. We analyzed invertebrate assemblages and environmental data, particularly measured pesticide concentrations in each stream, from five U.S. regions compiled within the United States Geological Surveys (USGS) regional stream-quality assessments (RSQA). We used boosted regression trees (BRTs) to assess the ability of single traits and TPGs to predict estimated pesticide toxicity. TPGs responded slightly more consistently than single traits across regions, but effective predictive models could be established for only one region. Then, we identified the most important traits and TPGs with the feature importance of the BRTs and used them as ecological indicators in a comparison with the fraction of EPT taxa and the SPEARpesticides indicator. We compared indicators by evaluating, for each, the performance, consistency, and specificity of its relationship to pesticide toxicity. The marginal effects of estimated pesticide toxicity were slightly higher and more consistent across regions for TPGs than single traits. However, in terms of performance, consistency, and specificity the SPEARpesticides indicator surpassed both TPGs and single traits. Given the limitations of current trait data, expert-defined indicators may be more effective than a data-driven approach using TPGs for assessing the impacts of pesticides on stream insect communities. Nonetheless, future developments of the TPG approach-particularly through the integration of physiological and genomic traits-hold promise for improving its predictive power.
The tomato leaf miner, Phthorimaea (Tuta) absoluta, Meyrick 1917 is recognized as a highly destructive pest, causing significant economic losses to crops in both greenhouse and open field environments across four continents: Asia, Africa, Europe, and South America. High genetic homogeneity among populations from various regions and countries indicates significant gene flow between P. absoluta populations, suggesting a lack of geographical barriers to dispersion. Furthermore, P. absoluta has developed resistance to insecticides due to target-site mutations or metabolic resistance, which enable the insect to withstand lethal doses of insecticides. To control this insect pest, the plant-mediated RNA interference (RNAi) is most promising host-induced gene silencing technique, utilized the plant's machinery to express double-stranded (dsRNA), which triggers the RNAi pathway in P. absoluta. Due to thermal tolerance, the P. absoluta has increased its area of invasion by 600 km per year over 9 years. Female P. absoluta releases pheromones that are recognized by males with a sophisticated olfactory circuit on their antenna. Pheromone binding proteins (PBPs) play a crucial role in mate recognition and attraction, and their expression peaks during courtship, specifically around 6:00 a.m. Given its potential to significantly alter the insect genome, clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (Cas9) offer a revolutionary strategy to control P. absoluta. Furthermore, this pest has developed remarkable adaptations to survive on unfavorable hosts by secreting specific proteins from its salivary glands that detoxify plant defenses. Insecticide resistance is likely the cause of field control failures of P. absoluta. Biological control, sex pheromone traps, and cultural control are the most promising approaches to address insecticide resistance resulting from these failures. Therefore, the implementation of integrated control programs and appropriate resistance management strategies is necessary to keep P. absoluta infestations under economic damage thresholds.
The purification of polysaccharides extracted from protein-rich matrices remains challenging due to strong carbohydrate-protein interactions. In this study, a sustainable and efficient deproteinization strategy was developed using a menthol-based deep eutectic solvent (DES) for purifying Protaetia brevitarsis seulensis polysaccharide (PBSP). Systematic DES screening and single-factor optimization were followed by multivariate modeling using response surface methodology (RSM) and an artificial neural network coupled with a genetic algorithm (ANN-GA). The ANN-GA model exhibited superior predictive accuracy with lower error metrics and higher R2 values than RSM. Under optimal conditions, the DES process achieved a polysaccharide retention rate (RR) of 95.30% and a deproteinization rate (DP) of 86.95%, corresponding to a comprehensive score (CS) of 91.12%. In contrast, the conventional Sevag method yielded a lower CS (68.53%) even after three extraction cycles and required approximately tenfold more solvent. Relative to PBSP, the DES-deproteinized polysaccharide showed approximately 61% higher total sugar and 75% lower protein contents, exhibiting superior purification efficiency compared with the Sevag-deproteinized polysaccharide. Structural characterization by molecular weight, FT-IR, NMR, and monosaccharide composition analyses demonstrated that DES treatment effectively removed proteins while preserving molecular integrity and the α/β-linked glucan backbone. Comparative evaluation across four edible insect polysaccharides from Oxya japonica, Tenebrio molitor, Zophobas morio, and Locusta migratoria, further confirmed the broad applicability of the developed DES process. Overall, this DES-based method provides a green, solvent-efficient, and scalable alternative for polysaccharide deproteinization, with strong potential for application to other insect-derived and protein-bound polysaccharides.
Frataxin is a conserved mitochondrial protein essential for cellular iron-sulfur (Fe-S) cluster biogenesis and oxidative balance, with its deficiency causing Friedreich's ataxia in humans. The hypomagnetic field (HMF), an environmental stressor known to influence oxidative stress and neurodevelopment, may interact with such inherent metabolic vulnerabilities. This study investigated whether HMF exposure exacerbates Fe-S homeostasis and oxidative disruption in a Drosophila melanogaster model of frataxin deficiency. Using synchrotron radiation-based X-ray fluorescence (SR-XRF) spectroscopy for in situ elemental analysis in live tissues, we found that HMF significantly altered iron distribution and content in a tissue-specific manner. In frataxin-silenced brains, HMF decreased iron distribution but increased total iron content, whereas in eyes it reduced iron content. Sulfur content decreased in frataxin-deficient eyes but increased in brains under HMF, though its spatial distribution was unchanged. Critically, HMF elevated reactive oxygen species (ROS) in frataxin-deficient brains. Transcriptomic analysis identified 202 differentially expressed genes under HMF in frataxin-silenced flies, including key regulators of iron metabolism and oxidative stress pathways. These findings demonstrate that HMF disrupts tissue-specific iron and sulfur homeostasis and intensifies oxidative stress in a frataxin-deficient insect system, underscoring its role as an environmental factor capable of aggravating metabolic fragility.
Mutualistic symbioses are potentially vulnerable to exploitation, particularly in hosts that acquire symbionts from the environment, where harmful exploiters inhabit. The independent evolution and persistence of intricate partner-choice mechanisms in many symbioses testify the threat by specialized exploiters of mutualisms, although only few have been documented in nature. We report here a lethal "Trojan horse" pathogen, Burkholderia sp. SJ1, exploiting the stinkbug-Caballeronia gut symbiosis. This bacterium resembles symbionts by using wrapping motility to traverse the host's sorting organ, inducing symbiotic organ morphogenesis and colonizing it. Unlike mutualists, however, it resists host digestion for nutrient acquisition, breaches the gut epithelium, and causes sepsis, rapidly killing the host. Colonization of the symbiotic organ is essential for its lethality. This case shows how pathogens can exploit mutualisms, highlighting the evolutionary pressures shaping partner-choice mechanisms and the fragility of even highly specialized mutualisms.
Aphis aurantii (Hemiptera: Aphididae) is a polyphagous insect pest that inflicts significant damage to diverse host plants, particularly within the Theaceae and Rutaceae, and transmits plant viruses such as citrus tristeza virus (CTV). These impacts highlight the need for genomic resources to support research on host adaptation and pest management. Here, we present the first high-quality chromosome-level genome assembly of A. aurantii. The assembled genome spans 347.32 Mb with a contig N50 of 40.10 Mb, and BUSCO assessment revealed a high completeness score of 96.85%. Using Hi-C scaffolding, 340.64 Mb of the genome was anchored into four pseudochromosomes, achieving a scaffold N50 of 90.37 Mb. Additionally, we annotated 16,881 protein-coding genes. This genome provides a reference resource for future investigations of genetic variation, host plant interactions, and polyphagy in aphids.
Climate change, land-use change, and intensified agricultural practices are reshaping agroecosystems, yet pest outbreak forecasts remain weak because links between individual-level mechanisms and spatiotemporal population dynamics are not well synthesized. We synthesize a framework in which global changes act through four proximate drivers: climate, host quantity/quality, top-down control, and management mortality, to reweight seven core processes i.e. development, reproduction, survival, diapause, aestivation, migration, and dispersal, thereby shaping within-season dynamics and cross-season carryover. A shared response architecture, signal sensing, neuroendocrine integration, and downstream physiological/molecular reprogramming, explains why warming typically accelerates development, shifts phenology, and increases voltinism, whereas extremes reduce survival and reproduction and intensify selection for resistance. We integrate physiological mechanisms to regulate spring-founding populations and distribution dynamics, highlighting destabilized diapause timing, heat-drought-driven aestivation, and wind-rain-mediated migratory redistribution. We identified key gaps that need to be resolved to parameterize cross-scale models and guide climate-smart pest management.
It was first hypothesized in 2009 that endogenous host reverse transcriptase (eRT) arising from retrotransposon elements in arthropod genomes would be critical for defense against viral pathogens in crustaceans and insects. This was not proven until 2016 in insects. In the succeeding 20 years, insect studies have confirmed the eRT role in antiviral defenses that result in tolerated, persistent infections via viral accommodation mechanisms (VAM). In insects, eRT transcribes viral RNA fragments into viral copy DNA (vcDNA) in both linear (lvcDNA) and circular (cvcDNA) forms. These, in turn, give rise to small interfering RNA (siRNA) fragments to feed the host RNA interference (RNAi) mechanism. At the same time, some vcDNA fragments are integrated into the host genome as endogenous viral elements (EVE), that can also give rise to siRNA. Studies with shrimp have progressed more slowly but results so far have mimicked those revealed with insects. Here we describe the presence and functional activity of a 5,383 bp-RT-related gene from a retrotransposon in the black tiger shrimp Penaeus monodon upon challenge with white spot syndrome virus (WSSV). It encompasses four conserved domains of protease (retropepsin), reverse transcriptase (RT-RNase H), transposase, and integrase as a single ORF of 1,248 deduced amino acids. The recombinant protein of a conserved 408-amino acid RT-RNase H domain, designated as Pm-eRT was expressed and found to possess reverse transcriptase activity. During WSSV infection, Pm-eRT was upregulated in shrimp hemocytes and pleopods and DNA/RNA hybrids in hemocytes were detected. The DNA/RNA hybrid signals were translocated from the plasma membrane to the nuclear membrane in the shrimp that survived WSSV infection with low viral loads. Pharmacological inhibition of Pm-eRT activity using azidothymidine (AZT) resulted in enhanced viral replication and accelerated shrimp mortality. Collectively, these findings support those described for insects, paving the way to further work on details of VAM in shrimp with the ultimate aim of developing genetically viral-tolerant breeding stocks for shrimp cultivation.
Tuta absoluta is a globally destructive invasive Solanaceae pest with widespread resistance to conventional insecticides, highlighting the urgent need for eco-friendly and sustainable control strategies. Catalase (CAT) is a key antioxidant enzyme that primarily scavenges intracellular hydrogen peroxide (H2O2) to maintain redox homeostasis. CATs have also been detected in the oral secretions (OSs) of some herbivorous insects, suggesting an additional extracellular role in manipulating plant defense during feeding making CATs innovative targets for pest management. Here, we systematically identified CATs in Tuta absoluta and evaluated their functions and suitability as RNA interference (RNAi) targets. Nine CATs were identified, with TaCAT1-TaCAT3 clustered phylogenetically with known insect OS-associated CATs. These genes showed high expression in larval heads and guts, induced by tomato feeding, indicating roles at the insect-plant interface. Silencing these CATs significantly impaired larval performance: individual double-stranded RNAs (dsRNAs) caused 69% mortality, while combined targeting (dsCAT-mix) achieved 80% mortality, accompanied by reduced pupation and smaller pupae. Functionally, CAT silencing induced tomato leaves H2O2 accumulation, and activated jasmonic acid pathway while suppressing salicylic acid signaling. These results demonstrate that CATs facilitate larval feeding by suppressing plant oxidative and hormonal defenses. Importantly, dsCAT treatments had no negative effects on predator Nesidiocoris tenuis, underscoring biosafety and specificity. These findings uncover a previously underappreciated role of CATs as putative OS effectors in a lepidopteran pest, validating them as effective, selective, and eco-friendly RNAi targets. This work provides mechanistic insight into insect-plant interactions and a practical foundation for sustainable RNAi-based strategies against Tuta absoluta. © 2026 Society of Chemical Industry.
Outbreaks of emerging and re-emerging diseases in both animals and plants are increasing due to climate change, globalization, land-use change, and agricultural intensification. While most pathogen monitoring programs focus on zoonotic threats, wildlife and other organisms in natural habitats can also serve as reservoirs and sentinels for pathogens of agricultural and ecological concern. Plant communities and the pathogens circulating within them are underrepresented in integrated disease monitoring frameworks. This study demonstrates how biodiversity and zoonosis monitoring programs conducted in protected habitats (tallgrass prairies and woodlands) across Illinois, together with insect specimens preserved in biorepositories, can be leveraged to improve knowledge of the identities and ecological associations of a wide range of potential pathogens. We developed an integrative workflow combining taxonomic identification, molecular screening, and epidemiological inference to detect vector-borne plant pathogens from archived insect material. Focusing on Hemiptera (Auchenorrhyncha), we screened specimens for phytoplasmas (Mollicutes), uncultured bacterial plant pathogens transmitted by sap-feeding insects, and characterized host-pathogen associations. At least three distinct phytoplasma strains were detected: 'Candidatus Phytoplasma asteris' (16SrI-B), 'Candidatus Phytoplasma pruni'-related strains (16SrIII), and 'Candidatus Phytoplasma sacchari'-related strains (16SrXI-H). The latter represents the first documented occurrence of a 16SrXI-H phytoplasma subgroup in Illinois. Overall, we identified five insect specimens harboring phytoplasmas across four preserved sites, all of which were previously unreported for insect-phytoplasma associations. These findings demonstrate the value of existing biodiversity and zoonosis monitoring infrastructures for proactive surveillance of plant pathogens and extending the One Health paradigm to explicitly include natural ecosystems.
The present study evaluated the biocontrol potential of soilborne Bacillus strains isolated from the largely unexplored soils of Khyber Pakhtunkhwa, Pakistan, against two globally significant lepidopteran pests, the white-marked tussock moth (Orgyia leucostigma) and the wax moth (Galleria mellonella). Among the isolates screened, Bacillus subtilis F1 exhibited high insecticidal activity, causing 74-77% mortality of larvae within 48 h when treated with whole-cell suspensions. In comparison, treatment with cell-free culture metabolites resulted in moderate mortality (20-23%), indicating that the primary insecticidal effect is largely cell-mediated, with extracellular metabolites contributing a secondary role. While the specific compounds responsible for the metabolite activity were not characterized in this study, previous reports suggested that Bacillus spp. may secrete proteins, enzymes, or secondary metabolites with insecticidal properties, which could explain the observed larval mortality. These findings demonstrated that infection, colonization, or combined cell-associated factors play a major role in larval mortality. To our knowledge, this is the first report of dual larvicidal activity of an indigenous B. subtilis strain against both O. leucostigma and G. mellonella. The present study also highlighted the presence of promising biocontrol agents in underexplored soil ecosystems, supporting their potential application in sustainable insect pest management. The online version contains supplementary material available at 10.1007/s13205-026-04779-y.