The escalating demand for sustainable pest management strategies has intensified research interest in entomopathogenic fungi (EPF), particularly regarding their taxonomic diversity and biocontrol potential in tropical ecosystems. However, only a limited number of comprehensive surveys that combine molecular phylogenetics with functional characterization have been conducted in Asia. In the present study, we collected mycosed insect cadavers from forest areas in Chiang Mai Province, northern Thailand. Morphological characteristics and multilocus phylogenetic analysis were conducted to facilitate proper species identification. Fungal isolates were further screened for proteolytic, chitinolytic, and lipolytic activities using a preliminary whole-plate assay. Further, a larval mortality bioassay using the mealworm beetle, Tenebrio molitor, was performed to confirm pathogenicity and assess the insecticidal potential of the fungal isolate under controlled laboratory conditions. In the field, seven insect cadavers infected with six different EPF species were collected. The collected fungal isolations were identified as Beauveria asiatica, B. bassiana, Clonostachys rogersoniana, Cordyceps blackwelliae, two strains of C. tenuipes, and Purpureocillium takamizusanense. For the enzymatic assay, C. tenuipes (MFLUCC 25-0373) has the highest chitinolytic activity, while P. takamizusanense (MFLUCC 25-0376) showed the highest proteolytic activity. In addition, Cl. Rogersoniana (MFLUCC 25-0379), C. tenuipes (MFLUCC 25-0373), and P. takamizusanense (MFLUCC 25-0376) showed higher and similar lipolytic activity. Notably, no single isolate in this study exhibited detectable activity for all three enzymes. Except for B. bassiana (MFLUCC 25-0375) and C. blackwelliae (MFLUCC 25-0377), which showed chitinolytic and proteolytic activity, respectively, all other isolates demonstrated activity for at least two of the three key enzymes. Most fungal isolates induced initial larval mortality by the third day post-exposure; however, virulence levels varied over time. Notably, no clear correlation was observed between enzyme activity and larval virulence, suggesting that fungal pathogenicity is a multifactorial process not merely dependent on extracellular enzymatic activity. Therefore, here, cumulative larval mortality was considered the primary criterion for candidate selection for future biocontrol formulation development, under which B. bassiana (MFLUCC 25-0375) was identified as the most suitable candidate, exhibiting the highest virulence.

The predatory bug Orius similis prefers to prey on the early-instar nymphs of Bemisia tabaci, while the parasitoid Encarsia formosa targets the late-instar nymphs. Consequently, complementary release strategies of both these natural enemies may be a promising tactic to increase downward pressure on B. tabaci populations and enhance current biological control strategies. This hypothesis was tested by examining different release ratios of natural enemies under different tempera-ture regimes within the laboratory and under more natural conditions using cage studies in the field. The results from this study indicated that when O. similis and E. formosa were released in ratios of 1:1, 1:2, and 1:3, the parasitism of E. formosa was higher than when it was released alone, whereas the predation of O. similis was lower than its solitary release. At 19, 22, 25, 28, and 31 degrees C, the responses of O. similis and E. formosa to B. tabaci nymphs were consistent with a type II functional response. The combined release of O. similis and E. formosa had the strongest control efficacy on B. tabaci nymphs at 28 degrees C. The effectiveness of different release ratios of O. similis and E. formosa on the control rate of B. tabaci nymphs was ranked as follows: 1 O. similis + 3 E. formosa > 1 O. similis + 2 E. formosa > 3 O. similis + 1 E. formosa > 2 O. similis + 1 E. formosa > 1 O. similis + 1 E. formosa > O. similis > E. formosa. Additionally, cage tests demonstrated that the combined release of O. similis and E. formosa effectively controlled the B. tabaci population. The 1:3 release ratio yielded superior control compared to other ratios. Therefore, when releasing O. similis and E. formosa in the field, it is recommended to do so at tem-peratures between 25 degrees C to 31 degrees C, with a ratio of 1:3, to significantly enhance the control of B. tabaci.

In the Multidimensional Management of Multiple Pests (3MP) theoretical framework, the bottom-up effects of soil fertilization on crop-insect pest-natural enemy multitrophic interactions can be leveraged to enhance pest control. Excessive nitrogen (N) fertilization is widespread in farming systems; however, its impact on pest control - particularly when multiple biocontrol agents are used simultaneously - remains poorly understood. Moreover, the implications of excessive N on soil-surface greenhouse gas emissions have not been fully explored. We conducted a series of mesocosm experiments using a Nicotiana tabacum - Myzus persicae - Aphidius gifuensis / Chrysoperla pallens system under semifield conditions. These trials assessed population-level effects and included behavioral and molecular assays to investigate predator feeding behavior. Despite detecting high rates of intraguild predation (IGP) in mesocosms - confirmed by midgut content molecular analysis due to spatial constraints - we found no significant difference in IGP rates between optimal and excessive N fertilization treatments. Behavioral assays revealed that the predator preferentially targeted and more effectively foraged unparasitized aphids. This behavior likely contributed to the greater aphid suppression observed in mesocosms where both parasitoid and predator were released together, compared to single-agent releases. Notably, excessive N input did not alter the combined top-down control of the two biocontrol agents on aphid populations. However, excessive N negatively affected tobacco growth and increased soil-surface emissions of nitrous oxide (N2O). These results underscore the importance of accounting for bottom-up fertilization effects in multitrophic interactions and highlight the need for balanced nitrogen management in integrated pest management (IPM) strategies.

The extensive reliance on synthetic pesticides has substantially enhanced agricultural productivity but has also generated persistent challenges related to environmental degradation, pesticide resistance, food safety, and human health. Biopesticides derived from microorganisms, plants, and biologically active natural compounds have emerged as sustainable alternatives that address these concerns while supporting ecologically sound pest management. This review critically evaluates the diversity of biopesticides, their formulation strategies, utilization prospects, and future directions, with emphasis on formulation science as a central determinant of efficacy, stability, and field performance. The article synthesizes recent advances in biopesticide formulations, including suspension systems, capsule suspensions, granular matrices, oil dispersions, and nano-enabled delivery platforms, highlighting their roles in enhancing persistence, controlled release, and target specificity. Particular attention is given to micro- and nanoencapsulation approaches that mitigate environmental sensitivity, extend shelf life, and facilitate integration into Integrated Pest Management programs. Regulatory challenges governing biopesticide commercialization are examined, underscoring the need for risk-proportionate and harmonized approval frameworks to accelerate innovation and market adoption. Emerging technologies, including synthetic biology, RNA interference, computational modeling, and microbiome-based strategies, are discussed as transformative tools that are reshaping biopesticide discovery and application. Overall, this review provides a consolidated perspective on the scientific and technological pathways required to advance biopesticides as reliable, scalable, and environmentally compatible components of sustainable crop protection systems.

[This corrects the article DOI: 10.3389/finsc.2025.1692096.].

Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is a major invasive pest threatening tomato production globally. Its high adaptability, rapid reproduction, and global spread from South America to Europe, Asia, and Africa have made it a serious challenge for growers. Conventional management relies heavily on chemical insecticides such as pyrethroids and neonicotinoids. However, the development of resistance, environmental concerns, and impacts on non-target organisms have limited their long-term effectiveness. Alternative strategies are increasingly needed. Cultural and physical controls, such as crop sanitation and traps, offer partial success. Biological control using natural enemies has shown promise but faces implementation challenges. Among emerging approaches, plant essential oils (PEOs) have gained attention as natural, biodegradable insecticides. PEOs exhibit diverse bioactivities—neurotoxic, repellent, antifeedant, and ovicidal—making them effective against T. absoluta. This review explores the pest’s biology, current control methods, and the potential of PEOs as part of sustainable pest management strategies. Future research should focus on improving PEO formulation, stability, and field performance to enable wider adoption in tomato pest management programs, supporting Zero Hunger, Responsible Consumption and Production.

Entomopathogenic fungi (EPF) are widely recognized as effective and environmentally sustainable biological control agents for a broad spectrum of agricultural insect pests. The increasing complexity and intensification of contemporary agricultural systems require the identification and development of more virulent and resilient fungal strains to ensure consistent pest suppression. In this study, ten EPF isolates representing Beauveria bassiana, Cordyceps javanica, and Metarhizium anisopliae were selected based on preliminary laboratory screens, and the insecticidal effectiveness of these isolates was tested against the mustard aphid, Lipaphis erysimi, along with the direct and interactive effects of plasma-activated water (PAW). Here, the PAW was generated using a surface dielectric barrier discharge (sDBD) plasma system, producing highly acidic water enriched with reactive oxygen and nitrogen species. Direct and daily application of PAW at concentrations of 50, 100, and 1,000 ppm resulted in complete aphid mortality within seven days at all tested concentrations. Conidial germination assays showed that lower PAW concentrations had less effect, whereas higher concentrations significantly inhibited germination. In certain isolates, the combined application of 100 ppm PAW and EPF produced greater aphid control than single EPF treatments. Overall, a single application of EPF achieved 87.67% aphid control, whereas the combined treatment achieved 99.67% mortality. Based on Lethal Time (LT50) values, C. javanica SLLC-Cj 24 (single application average 6.93 days; combined application average 6.60 days) was the most virulent strain, followed by SLLC-Cj 11 (single application average 7.17 days; combined application average 6.65 days). Among the tested fungal strains, B. bassiana SLLC-Bb 12 demonstrated the strongest synergistic interaction with PAW, followed by C. javanica SLLC-Cj 1. These findings demonstrate that PAW possesses intrinsic insecticidal activity and can enhance overall mortality when integrated with EPF. However, isolate- or strain-specific interactions may influence mortality and infection rates, highlighting the need to select fungal cultures, optimize concentration, and exposure parameters for synergistic biocontrol strategies.

Optimizing the growth performance of black soldier fly ( Hermetia illucens L.) (Diptera: Stratiomyidae) larvae (BSFL) remains challenging due to limited understanding of the influence of genetic strain. Moreover, the effects of strain cannot be accurately assessed without controlling for confounding factors such as rearing density and diet fermentation variables often overlooked in previous studies. In this study, we evaluated the growth performance and density sensitivity of BSFL strains with varying degrees of inbreeding, inferred from the frequency of wild-type line introductions. Larvae were reared at defined densities using a standardized, fermentation-reduced diet to minimize microbial and nutritional variability. Daily changes in body width were modelled using logistic growth functions to estimate key parameters: asymptotic growth limit ( L ), inflection point ( m ), and growth rate ( k ). These parameters, along with fresh weight, dry weight, and body mass index at prepupa stage, were compared among strains. Correlations between density and growth traits were also analysed. Additionally, fatty acid and amino acid profiles were assessed. Results showed that strains with frequent wild-type introductions resulted in enhanced growth under low-density conditions but were more sensitive to crowding, whereas domesticated strains demonstrated greater tolerance to high density. However, highly domesticated strains exhibited significantly reduced body size and dry weight. Total fat and amino acid content remained consistent across treatments, with minor strain-specific variations on specific amino acid. These findings highlight the importance of genetic strain, density control, and standardized protocols in improving BSF rearing efficiency and ensuring reproducible research outcomes, which contributes to Sustainable Development Goal (SDG) 2, Zero Hunger, through its implications for sustainable insect-based protein production.

Tomato leaf miner, Tuta absoluta, is a major pest responsible for significant yield losses in tomato cultivation worldwide. Increasing resistance to chemical insecticides and growing environmental concerns have underscored the need for eco-friendly pest management alternatives aligned with sustainable development Goals (SDGs) such as zero hunger and responsible consumption and production. This study evaluated the insecticidal potential of Annona squamosa seed extracts prepared using five different solvents: acetone, ethyl acetate, ethanol, methanol, and hexane. Their toxicity was tested against T. absoluta eggs and larvae. Bioassays revealed that the hexane extract, at the highest concentration (250 ppm), completely suppressed egg hatchability (0%) up to 120 h. The same concentration exhibited larvicidal activity, causing 46.66% at 24 h and 73.33% mortality at 48 h post-treatment. The lowest LC₅₀ and LC₉₀ values were observed in the hexane extract: 155.04 and 344.06 ppm at 24 h, and 85.49 and 169.43 ppm at 48 h, respectively. Enzymatic assays indicated a decrease in catalase (CAT) activity and an increase in glutathione S-transferase (GST) activity in T. absoluta larvae 24 h after treatment with the hexane extract. A non-target assay on Eudrilus eugeniae showed 5.33% minimal mortality at the highest concentration, compared to 96.66% mortality in Imidacloprid treatment and 0% in control. GC-MS analysis identified two major compounds, E-11-hexadecenoic acid ethyl ester (25.18%) and 9,12-octadecadienoyl chloride (Z, Z) (15.23%), which may be involved in the insecticidal activity. Molecular docking studies demonstrated strong binding affinities of E-11-hexadecenoic acid ethyl ester and 9,12-octadecadienoyl chloride (Z, Z) with the target insect enzyme acetylcholinesterase (AChE), suggesting their potential mode of action. These findings indicate that the hexane extract of A. squamosa seeds contains promising bioactive molecules with significant insecticidal potential. This research highlights the potential of plant-derived insecticides for environmentally safe and sustainable management of T. absoluta, reducing dependence on synthetic chemicals and promoting sustainable tomato production, good health and well-being and responsible consumption and production.

Introduction:Black soldier fly larvae (BSFL) have gained increasing attention as a sustainable alternative protein source for animal feed, particularly when reared on organic by-products. This study evaluated the nutritional composition of BSFL reared on different organic substrates to support sustainable feed production. Methods:BSFL were reared on five organic substrates: chicken feed, pig feed, soy milk residue, coconut press cake, and perilla cake. Larvae were cultivated in metal trays (23 × 15 cm) for 2-4 weeks under controlled conditions (28 ± 2 °C; 65 ± 5% relative humidity). At the prepupal stage, larvae were harvested and analyzed for dry matter, crude protein, crude fiber, ether extract, ash, growth performance, and fatty acid profiles. Results:Significant differences in nutritional composition were observed among substrates. Crude protein content was high in larvae fed chicken feed (50.55 ± 0.07%), pig feed (52.10 ± 0.14%), soy milk residue (52.15 ± 0.78%), and perilla cake (47.20 ± 0.00%). Crude fiber was highest in larvae fed soy milk residue (7.19 ± 1.48%) and perilla cake (5.38 ± 0.25%). Favorable growth performance was observed, with larvae reaching 0.22 ± 0.01 g in weight, 20.38 ± 0.36 mm in length, and 5.08 ± 0.05 mm in width. Fatty acid analysis revealed substantial levels of saturated and unsaturated fatty acids, including palmitic, oleic, and linoleic acids. Larvae reared on coconut press cake showed the highest saturated fatty acid content (74.91 ± 0.03%), while those fed soy milk residue exhibited the highest oleic (26.68 ± 0.06%) and linoleic acid (38.44 ± 0.07%) contents, resulting in increased polyunsaturated fatty acids (38.57 ± 0.03%). Discussion:The findings demonstrate that organic by-products commonly available in Thailand are suitable substrates for BSFL production and significantly influence larval nutritional quality. These substrates enable the production of nutrient-rich, cost-effective, and sustainable insect-based feed, contributing to responsible consumption, waste valorization, and food security.

Antithrombotic proteins of medicinal leeches-originally characterized from salivary-gland secretions-suppress coagulation, platelet activation and fibrinolysis at the feeding site, but the contribution of alternative transcript processing to this repertoire remains poorly documented. Here we used four H. manillensis RNA-seq libraries (THA-Hm1–THA-Hm4), whose species identity and overall sequencing performance were characterized in our companion study, to determine how many antithrombotic loci produce multiple structurally distinct transcript isoforms and how such variation extends into coding capacity. Reference-guided assembly merged into a unified transcript set identified 38 transcript-supported candidates in addition to the 72 catalogued reference genes. When candidate transcripts were grouped by parent MSTRG locus and ranked by isoform count, six loci carried three or four structurally distinct models: MSTRG.4545, MSTRG.5212, MSTRG.3310, MSTRG.5709, MSTRG.5200 and MSTRG.5713. Transcript-length variation alone produced isoforms at three of the six loci, whereas at MSTRG.5200, MSTRG.5709 and MSTRG.5713 the variation extended into the coding sequence, producing predicted proteins ranging from 49 to 1,159 amino acids. These results reveal localized isoform complexity at a small set of antithrombotic loci and define a focused candidate set for downstream domain annotation, splice-junction visualisation and functional validation.

The tomato leaf miner, Tuta absoluta, is a devastating pest that causes severe yield losses in tomato cultivation, threatening food security (SDG 2: Zero Hunger). Reliance on chemical insecticides has led to resistance development and environmental concerns, highlighting the need for sustainable pest management strategies that promote responsible consumption and production (SDG 12). In this context, the present study focused on the green synthesis of silver nanoparticles (AgNPs) using hexane extracts of Annona squamosa seeds and evaluated their potential as an eco-friendly insecticidal agent. The synthesized AgNPs were characterized physiochemically and structurally using UV–Vis spectroscopy, FTIR, XRD, EDX, and SEM analyses. The UV–Vis spectrum confirmed nanoparticle formation; FTIR analysis identified functional groups responsible for reduction and stabilization; XRD revealed the crystalline nature; and SEM/EDX showed spherical morphology with elemental silver signals. Insecticidal bioassays against T. absoluta demonstrated significant larval mortality of up to 96.66% within 48 h at 90 ppm. Biochemical assays revealed alterations in detoxification enzymes, with catalase (CAT) and glutathione S-transferase (GST) showing enhanced activity after 24 h of exposure, suggesting oxidative stress induction and enzyme modulation. Safety evaluation on the non-target organism Eudrilus eugeniae revealed low toxicity of 8.33% after  24 h when treated with nanoparticles synthesized from the hexane extract, highlighting environmental safety and supporting life on land and biodiversity conservation (SDG 15). Furthermore, molecular docking studies indicated strong binding affinities of bioactive phytochemicals from A. squamosa with key insect enzyme targets, such as acetylcholinesterase (AChE), supporting their role in nanoparticle-mediated toxicity. This study demonstrates the insecticidal efficacy and mode of action of A. squamosa-derived AgNPs. The findings support their potential as safe, sustainable nano-pesticides for managing T. absoluta, contributing to sustainable agriculture (SDG 2), promoting innovative, eco-friendly technologies (SDG 9), and fostering environmental sustainability (SDG 12, 15).

Soil organic fertilization and biological control-key components of the Multidimensional Management of Multiple Pests (3MP) theoretical framework-are known to influence insect pest populations. To evaluate their combined impact, we employed the Performance-Economy-Environment (PEE) multi-criteria assessment, which considers pest control efficacy, crop yield and quality, and environmental outcomes, to develop innovative IPM packages. However, such an integrated assessment of packages involving both organic fertilization and biological control has been lacking. We conducted a semi-field experiment using the Solanum lycopersicum-Bemisia tabaci-Encarsia formosa system under two fertilization regimes: full chemical fertilization (CF) and partial replacement with organic manure (COF). We found that B. tabaci adult and nymph abundance was significantly lower under the COF regime when E. formosa was present, although parasitism rates remained similar across both fertilization treatments-suggesting comparable top-down control. This implies that the COF regime exerted negative bottom-up effects on B. tabaci populations. In addition to improved pest suppression, the COF regime enhanced tomato yield and fruit quality, as evidenced by higher lycopene and soluble solids content and lower nitrate accumulation. Environmentally, the COF regime improved soil fertility-indicated by increased soil organic matter and organic carbon-and reduced nitrous oxide (N2O) emissions by 16.25%, a key greenhouse gas. This first application of the PEE assessment framework to an IPM package integrating organic fertilization and biocontrol highlights the ecological, agronomic, and economic benefits of such sustainable pest management strategies.

AIMS:The challenges of mass production and environmental stress affect the application of entomopathogenic fungi (EPF). Therefore, this study attempts to develop a procedure based on EPF molecular markers to facilitate the EP selection. METHODS AND RESULTS:In this study, three Beauveria bassiana isolates and six Metarhizium spp., were evaluated for conidial production and environmental stress tolerances. The expression levels of six phenotypic related genes were analyzed to assess correlations with conidial production and stress tolerance in two phenotypic different isolates of B. bassiana and Metarhizium spp. In B. bassiana isolates, both slt2 and catA showed strong correlations with conidial production and thermotolerance. Additionally, slt2 and hog1 were associated with osmotic stress tolerance and oxidative stress tolerance, respectively. However, the correlation between phenotypes and gene expression trends was not consistent in Metarhizium spp. Further validation using three M. pinghaense isolates indicated that hog1 was strongly correlated with conidial production, while mbf1, slt2, and cag8 were associated with thermotolerance, oxidative stress tolerance, and osmotic stress tolerance, respectively. CONCLUSIONS:These findings suggest the existence of intraspecies conservation in phenotypic mechanisms and offer a strategy for the rapid selection of promising EPF strains.

Tuta absoluta (Meyrick) has emerged as a significant pest of solanaceous crops, particularly tomatoes. Although its host range includes other plants within the Solanaceae family, this manuscript presents the first evidence of T. absoluta infesting and causing damage to Solanum indicum (Indian nightshade). Observations were conducted under greenhouse conditions, where pest-induced foliar damage caused by larval feeding was documented. These findings expand the known host range of T. absoluta, highlighting the potential risks of additional solanaceous crops and informing future pest management strategies.

Bemisia tabaci (Gennadius) is among the most serious threats to global agricultural production. Biological control of B. tabaci using the parasitoid Encarsia formosa Gahan is effective on various crops. However,the biological control of B. tabaci could be affected by various abiotic factors. Understanding the effects of temperature on parasitoid efficacy is one of the key ascpects in this case. In this study, the functional response, searching efficiency, and parasitism preference of E. formosa on early and late nymphal instars of B. tabaci at different temperatures were investigated. We found that the functional response conformed to the type II model at 19, 22, 25, 28 and 31 degrees C. The parasitoid efficiency of E. formosa toward nymphal B. tabaci was the highest at 28 degrees C. Searching efficiency of E. formosa decreased with the increase in host density. The Hassell-Varley interference model simulation indicated intraspecific interference in E. formosa. E. formosa preferred to parasitize late instar nymphs of B. tabaci compared to the early instar nymphs. In conclusion, E. formosa releases at 25-31 degrees C with a parasitoid-to-host ratio of 1:50 is considered optimal to control B. tabaci in field, which minimizes costs and maximizes parasitism rate.

The current study aimed to isolate Beauveria brongniartii conidia from forest soils, identify the fungus, and evaluate its effectiveness on the eggs, larvae, pupae, and adults of Spodoptera litura. Insect mortality rates were recorded every 3, 6, 9, and 12 days. The identification of entomopathogenic fungi was carried out using molecular techniques, including PCR, DNA sequencing, and molecular markers, to detect species-specific 18 S rDNA genetic sequences, all performed under aseptic conditions. The results indicated that higher conidia concentrations (2.7 × 109 conidia/mL) exhibited greater virulence, with eggs showing a mortality rate of 98.66%, followed by larvae 96%, adults 90.66%, and pupae 77.33% after 12 days. Probit analysis revealed minimal LC50 and LC90 values: eggs (5.5 × 102; 1.0 × 106 spores/mL), larvae (8.2 × 102; 1.2 × 107 spores/mL), pupae (9.6 × 104; 7.3 × 1010 spores/mL), and adults (1.0 × 103; 2.0 × 108 spores/mL). The total hemocyte counts and detailed observational results revealed that B. brongniartii induces cellular breakdown and cell lysis in S. litura larvae by producing enzymes that degrade the cuticle and cell membranes. Earthworm bioindicator studies showed minimal effects from B. brongniartii conidia compared to controls, while chemical treatments resulted in 96% mortality at 100 ppm. Histopathological examinations revealed no significant differences in gut tissue between earthworms treated with fungal conidia and those in the control group, unlike the substantial damage caused by chemical treatments. Biochemical analysis revealed significant alterations in enzyme activity, including reduced levels of phosphatase and catalase, as well as increased levels of lipid peroxides and superoxide dismutase. This study highlights the effectiveness of B. brongniartii in controlling S. litura, demonstrating its potential as a viable biocontrol agent and promoting eco-friendly alternatives to chemical pesticides, with no risk to non-target species or the environment.

Aedes albopictus, known as the Asian tiger mosquito, is a significant vector for dengue fever, chikungunya, zika virus, yellow fever. Current control methods rely on chemical insecticides, which face challenges such as resistance, environmental harm, and impact on non-target species Eudrilus eugeniae and Artemia salina. This study evaluates the toxic effects of biogenic copper nanoparticles (CuNPs) synthesized using Metarhizium robertsii intracellular extract obtained from our previous research. The CuNPs were tested against A. albopictus and non-target species at 24 and 48 hours post-treatment. Results demonstrated that entomopathogenic fungi-derived CuNPs exhibited potent mosquitocidal activity, resulting in 97.33% mortality in larvae, 93.33% in pupae, and 74.66% in adults at 48 hours post-treatment. The CuNPs derived from M. robertsii showed lower LC50 values of 74.873 mg/L in larvae, 76.101 mg/L in pupae, and 136.645 mg/L in adults at 48 hours post-treatment. Additionally, 12 hours post-treatment, catalase (an antioxidant enzyme) activity decreased 1.5-fold in a dose-dependent manner, while glutathione S-transferase (a detoxification enzyme) activity increased 7.8-fold. CuNPs demonstrated lower toxicity to non-target species, with 24% mortality in A. salina and 24.44% mortality in E. eugeniae at 24 hours post-treatment. The LC50 values were 634.747 mg/L for A. salina and 602.494 mg/L for E. eugeniae at 24 hours post-treatment. These findings indicate that entomopathogenic fungi-derived CuNPs are a promising, target-specific candidate for controlling A. albopictus at various life stages (larvae, pupae, and adults).