Nanoemulsion based on essential oil of Mentha piperita in the sustainable management of Oligonychus ilicis McGregor, 1917 (Acari: Tetranychidae)

Nanoemulsion based on essential oil of Mentha piperita in the sustainable management of Oligonychus ilicis McGregor, 1917 (Acari: Tetranychidae) | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Nanoemulsion based on essential oil of Mentha piperita in the sustainable management of Oligonychus ilicis McGregor, 1917 (Acari: Tetranychidae) Vanessa Racaneli Sian, Gustavo Pazolini Stein, Ana Beatriz Mamedes Piffer, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9260778/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 13 You are reading this latest preprint version Abstract The red mite, Oligonychus ilicis McGregor, 1917 (Acari: Tetranychidae), is a major pest of coffee crops ( Coffea canephora ), causing significant damage to plantations. In this context, essential oils, particularly when formulated as nanoemulsions, emerge as a promising alternative for pest management due to their high efficacy and low environmental toxicity. This study explores the acaricidal potential of a nanoemulsion based on Mentha piperita essential oil against O. ilicis . For this study, an analysis of the main chemical constituents of the essential oil (EO) was first carried out, and a nanoemulsion based on peppermint essential oil was formulated using the low-energy emulsification method, with specific surfactants employed to stabilize the particles. The proposed formulation remained stable over time, and subsequently, the toxicity of the nanoemulsion was evaluated through laboratory bioassays involving the direct application of different concentrations (0.025, 0.040, 0.060, 0.10, 0.15, and 0.25% [mL mL⁻¹]) to adult females of O. ilicis . The results revealed a dose-dependent mortality, with mortality rates exceeding 90% for O. ilicis at concentrations as low as 0.040%. The LC₅₀ and LC₉₀ values were estimated at 0.0257 and 0.0424% (mL mL⁻¹), respectively. These findings demonstrate that the peppermint essential oil-based nanoemulsion exhibits strong acaricidal activity against the red coffee mite, O. ilicis . Given its efficacy and potential environmental safety, this formulation represents a promising and sustainable tool for the integrated management of this key pest in coffee production systems. alternative pest control biopesticide Coffea canephora phytochemicals red coffee mite Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 1 Introduction The red mite, Oligonychus ilicis McGregor, 1917 (Acari: Tetranychidae), is a phytophagous arthropod of major economic importance and is described as one of the key pests of Conilon coffee ( Coffea canephora Pierre & Froehn) (Piffer et al., 2023 ). This species preferentially colonizes the adaxial surface of leaves, where it pierces epidermal cells to feed, extracting part of the cellular contents and imparting a characteristic bronzed appearance to the foliage. In association with its presence, a delicate web produced by the mites themselves can accumulate dust and debris, giving leaves a dirty appearance (Ozman et al., 2024 ). Prolonged dry periods with water deficit are favorable for mite outbreaks and may lead to severe defoliation. In young plantations, the damage caused by this pest reduces photosynthetic capacity, compromises vegetative growth and vigor, and can directly affect crop survival (Lopez and Liburd, 2020 ; Marucci et al., 2024 ). According to the Ministry of Agriculture, Livestock and Supply (MAPA), there are approximately 70 products registered for the control of the red coffee mite, of which only two are classified as environmentally safe (Agrofit, 2025). Most of these products contain avermectin-based active ingredients, which not only cause mortality and reduced reproduction in soil invertebrates but may also induce phytotoxic effects in plants (Souza and Guimarães, 2022 ). Moreover, synthetic chemicals have adverse effects on non-target organisms and aquatic resources (Carducci et al., 2019 ). To date, there are no officially registered biological products for the management of O. ilicis , highlighting the urgent need for research and the development of sustainable and environmentally safer alternatives (Agrofit, 2025). Essential oils (EOs) are metabolites produced through the secondary metabolism of plants as a defense mechanism against arthropods. Their bioactivity is associated with phytochemical classes such as terpenoids, alkaloids, flavonoids, steroids, saponins, and tannins, many of which are naturally toxic to pests (Richardson et al., 2015 ; Divekar et al., 2022 ). Among the plant species with reported acaricidal properties, peppermint ( Mentha piperita ) is noteworthy (Souza et al., 2022 ). Peppermint is a perennial aromatic herb containing important compounds such as menthol, menthone, limonene, isomenthone, menthyl acetate, carvone, β-pinene, and 1,8-cineole, all of which have demonstrated activity against pest organisms (Richardson et al., 2015 ; Stringaro et al., 2018 ; Divekar et al., 2022 ). However, under environmental conditions, EOs are chemically unstable and undergo rapid degradation (Zhang et al., 2022 ; Albuquerque et al., 2022 ). Nanoemulsions have attracted increasing attention in agriculture due to their ability to stabilize lipophilic active ingredients, provide controlled release and prolonged bioactivity, and enhance the chemical stability of formulations by preventing phase separation (Ghosh et al., 2014 , Divekar et al., 2022 ). Furthermore, these formulations represent a promising alternative to synthetic chemicals, as they tend to exert lower environmental impact (Annadurai et al., 2024 ; Modafferi et al., 2025 ). In this context, the objective of this study was to characterize the chemical composition of M. piperita essential oil, formulate a peppermint oil-based nanoemulsion, and evaluate its acaricidal effect against O. ilicis under laboratory conditions. 2 Materials and Methods The experiments were conducted at the Laboratory of Entomology and Agricultural Acarology of the Federal Institute of Education, Science and Technology of Espírito Santo – Itapina Campus (IFES – Itapina Campus), located in the municipality of Colatina, at the geographic coordinates 19°29'52.7"S and 40°45'38.5"W (Fig. 1 ). The bioassays were carried out in climate-controlled chambers at 25 ± 1°C, 70 ± 10% relative humidity, and a 12 h photoperiod. 2.1 Essential oil acquisition and chemical composition analysis The essential oil was commercially obtained from Ferquima Indústria e Comércio Ltda (Batch 218). The chemical characterization of the essential oil was performed by gas chromatography coupled with mass spectrometry (GC-MS), using an Agilent 7890B gas chromatograph coupled to a 5977A MSD mass selective detector, operating with electron impact ionization at 70 eV. Compound separation was carried out using an HP-5 capillary column (30 m × 250 µm × 0.25 µm). The injector and detector temperatures were set at 290°C and 310°C, respectively. The oven temperature program consisted of an initial temperature of 40°C, followed by a heating ramp of 5°C/min up to 280°C, and a second ramp of 15°C/min up to a final temperature of 310°C. For the calculation of Kovats retention indices (KI), a standard mixture of n-alkanes ranging from C10 to C40 was used. Compound identification was based on comparison of the obtained mass spectra with the NIST library data, as well as correlation of retention indices and spectral patterns with reference data from the literature (Adams, 2017). 2.2 Formulation and physicochemical characterization of the Mentha piperita nanoemulsion The peppermint essential oil-based nanoemulsion was obtained using the low-energy emulsification method, with adaptations (McClements, 2012 ). Initially, 24 combinations varying in hydrophilic-lipophilic balance (HLB) values and surfactant ratios were tested to identify the most stable formulation. The oil phase (essential oil + surfactant) was homogenized by vortexing, after which the aqueous phase (distilled water) was slowly added under continuous stirring until reaching a final mass of 1 g (50 mg/mL). The most stable M. piperita nanoemulsion was achieved at HLB 15, using 5% essential oil, 20% surfactants (Polysorbate 20 and Sorbitan monooleate), and 75% water. Subsequently, the formulation was diluted to concentrations of 2.5%, 1%, 0.5%, and 0.2% essential oil. Nanoemulsion stability was monitored on days 0, 17, and 34 after preparation through visual assessment of macroscopic characteristics, following certain criteria (Ortiz-Zamora et al., 2020 ; Lima et al., 2021 ) (Fig. 2 ). The presence of a continuous phase and the Tyndall effect was observed by a bluish coloration under incident light, indicating the formation of nanostructures in the formulations. The dilutions that remained macroscopically stable (1, 0.5, and 0.2%) were subjected to hydrodynamic diameter, polydispersity index (PDI), and zeta potential analyses. FT-IR analyses were performed using a Spectrum Two spectrometer equipped with a diamond ATR (Attenuated Total Reflection) accessory. The PerkinElmer Spectrum software was used for spectral processing. Spectra were acquired with 32 scans per spectrum of the surfactants, in triplicate, within the mid-infrared range of 4000–400 cm⁻¹, with a spectral resolution of 8 cm⁻¹, adapted (Carvalho et al., 2023 ). Colloidal characterization was carried out by dynamic light scattering (DLS) using a Litesizer 500® (Anton Paar) to determine the average hydrodynamic diameter, polydispersity index (PDI), and zeta potential, with a 1:400 (v/v) dilution in distilled water. Measurements were taken on days 1, 17, and 34 after preparation at concentrations of 1%, 0.5%, and 0.2%, with up to 1000 runs per sample. Based on stability analyses and the maintenance of physicochemical parameters observed at concentrations of 0.5% and 0.2%, lower dilutions were selected for the bioassays against mites. Thus, concentrations of 0.025%, 0.040%, 0.060%, 0.10%, 0.15%, and 0.25% were used in the biological tests, obtained from progressive dilution of the previously stabilized formulation. 2.3 Rearing of Oligonychus ilicis Mites were collected from a coffee plantation at IFES – Itapina Campus, where no chemical products had been applied, and subsequently reared under laboratory conditions following an adaptation of the technique (Reis et al., 1997 ). In the same plantation, leaves were collected, washed in distilled water, and disinfected in a sodium dichloroisocyanurate solution (1:10). Petri dishes (14.0 × 1.5 cm) were prepared as rearing arenas, with moistened cotton placed at the bottom and around the coffee leaf to maintain leaf turgor and prevent mite escape. The arenas were maintained in climate-controlled B.O.D. chambers at 25 ± 1°C, 70 ± 10% relative humidity, and a 12 h photoperiod, with weekly maintenance to ensure colony preservation under laboratory conditions. 2.4 Laboratory bioassays with essential oil-based nanoemulsions against Oligonychus ilicis For the bioassays, O. ilicis females were standardized to obtain adult individuals of uniform age. Arenas were assembled in Petri dishes (10.0 × 1.2 cm) containing coffee leaf discs (4 cm in diameter), with moistened cotton placed around the discs to maintain leaf turgor. Ten females were carefully transferred to each dish using a fine-bristle brush. The nanoemulsion was tested at concentrations of 0.025, 0.040, 0.060, 0.10, 0.15, and 0.25% (mL mL⁻¹), obtained from the base formulation. Each treatment consisted of 10 replicates. The control treatment consisted of distilled water with Tween® 80 (0.05%) as a surfactant. Application of the solutions to the mites was performed using an airbrush calibrated at a constant pressure of 15 psi, delivering 1 mL of solution per replicate. Mite mortality was assessed at 12, 24, 36, 48, 60, and 72 hours after spraying. Individuals were considered dead when they failed to move after gentle stimulation with the brush bristles. 2.5 Statistical analysis Data were subjected to Probit regression analysis, and the resulting equations were used to estimate LC₅₀ and LC₉₀ values. Survival data were analyzed using a Cox proportional hazards model, and treatment contrasts were evaluated with the emmeans function of the corresponding R package (Lenth, 2023). All statistical analyses were performed using R software, version 4.3.2 (R Core Team, 2023). 3 Results 3.1 Chemical composition of peppermint oil by gas chromatography–mass spectrometry Twelve compounds were identified, accounting for 99.4% of the total peak area (Table 1). Among them, the major constituents were isomenthol and isomenthone, which represented 67.4% of the oil (Fig. 3). The presence of monoterpene ketones, monoterpene alcohols, monoterpene ethers, and hydrocarbons was detected. When correlating the absorption bands observed in the FT-IR spectra with the compounds identified in the essential oil of M. piperita (Table 2), a large number of bands below 1500 cm⁻¹ were observed, which is associated with the diversity of functional groups among the substances identified by GC-MS. In addition to the bands related to alkane and olefin vibrations, sharp and intense peaks were observed, corresponding to the C=O stretching vibration at 1750–1700 cm⁻¹, the C–O stretching vibration at 1050 cm⁻¹, and the asymmetric C–O–C stretching vibration at 1280–1230 cm⁻¹. Table 1 Major chemical constituents of Mentha piperita essential oil identified by GC-MS analysis. Compound name Relative area (%) Menthol (isomer) 42,079 Menthone 25,323 Menthol (neoisomer) 8,759 Menthyl acetate 7,616 1,8-Cineole 6,49 Limonene 2,544 Caryophyllene oxide 1,683 β-Pinene 1,258 Caryophyllene (E) 1,159 Pulegone 1,091 α-Pinene 0,996 Piperitone 0,433 3.2 Macroscopic stability, hydrodynamic size, and zeta potential analysis The peppermint essential oil-based nanoemulsion at different concentrations was characterized in terms of droplet size, hydrodynamic diameter, polydispersity index (PDI), zeta potential, and particle size distribution peaks over time. The use of the low-energy input method enabled the development of highly stable nanoemulsions, with droplet sizes in the nanometric range, low PDI values, and good overall stability (Fig. 4). Initially, all concentrations exhibited stability. The 1% dilution, although displaying a whitish appearance, did not undergo phase separation and retained nanometric droplet size; however, it did not remain stable over time (Fig. 4). In contrast, the 0.2% and 0.5% dilutions remained stable until the final day of analysis, without significant differences in hydrodynamic diameter between them. Furthermore, the zeta potential and polydispersity index values demonstrated stability (Fig. 4). The dilutions exhibited hydrodynamic diameters of 59.25 nm and 58.64 nm, respectively. Based on these data, and considering the visual homogeneity observed in the samples, it can be inferred that even lower dilutions, such as those used in the bioassays (0.025% to 0.25%), exhibit equivalent physicochemical behavior, maintaining the integrity of the nanostructured system. The stability observed in the 0.5% and 0.2% samples, together with the absence of flocculation or phase separation and the preservation of colloidal properties, supports this inference, despite the instrumental limitation in directly measuring such lower concentrations. 3.5 Toxicity effect of the nanoemulsion based on Mentha piperita essential oil An acaricidal effect of the peppermint essential oil-based nanoemulsion was verified, with adult mortality of O. ilicis increasing in a dose-dependent manner. The lethal concentrations for 50% and 90% of the population were estimated at 0.0257 and 0.0424 (mL·mL⁻¹), respectively (Fig.5). In the survival analysis of O. ilicis , the highest mortality rates occurred within 12 hours after direct application on the individuals, reaching up to 100% mortality at concentrations of 0.15% and 0.25% (Fig. 6). 4 Discussion Nanoemulsions, due to their nanometric-scale droplets, are structurally more susceptible to destabilization by Ostwald ripening (Koroleva and Yurtov, 2021 ). However, this same reduced size favors their kinetic stability by minimizing phenomena such as flocculation, coalescence, and gravitational separation, particularly in well-balanced formulations (Oca-Ávalos et al., 2017 ). With lower hydrodynamic diameter and polydispersity index values, the results obtained in this study are consistent with the standards established by some researchers, indicating the development of a stable and promising system (McClements, 2012 ). Such stability favors the controlled and prolonged release of the phytochemical compounds present in the essential oil, contributing to the lethal effects observed in the bioassays (Ghosh et al., 2023 ; Ayllón-Gutiérrez et al., 2024 ). The toxicity of essential oils may be directly related to the diversity and concentration of their active constituents, which vary according to agronomic, edaphoclimatic, and methodological factors, such as soil type, extraction method, and plant phenological stage, among others (Khan et al., 2023 ; Abd-Elnabi et al., 2025 ). The chemical analysis of M. piperita essential oil revealed the presence of compounds belonging to three main chemical groups: hydrocarbon monoterpenes, oxygenated monoterpenes, and oxygenated sesquiterpenes. Among these, oxygenated monoterpenes stand out, particularly menthol and menthone, which emerged as the major constituents. These compounds have been extensively studied due to their recognized insecticidal and repellent properties (Baker et al., 2023 ). Menthol is capable of inhibiting the enzyme acetylcholinesterase (AChE), leading to the accumulation of acetylcholine at synapses, resulting in nervous hyperexcitation, paralysis, and insect death (Finetti et al., 2021 ; Liu et al., 2022 ). Menthone, on the other hand, although also presenting this effect, has its activity more associated with the modulation of ion channels, especially sodium channels and GABA receptors, altering nerve impulse conduction and causing motor incoordination (Gad et al., 2022 ; Souza et al., 2022 ; Wu et al., 2023 ; Santos et al., 2024 ). Research has shown that the essential oils of M. piperita exhibit residual contact toxicity and exert a repellent and food deterrent effect on third-instar larvae of the diamondback moth ( Plutella xylostella L., Lepidoptera: Plutellidae)(Koundal et al. , 2024; Paz et al. , 2025). Additionally, the methanolic extract of M. piperita is demonstrated as effective as a larvicide and oviposition inhibitor for this pest (Afiunizadeh et al., 2022 ; Koundal et al. , 2024; Paz et al. , 2025). Also, researchers reported effective control of the fall armyworm ( Spodoptera frugiperda (Lepidoptera: Noctuidae)) using peppermint oil, reinforcing its potential as a broad-spectrum bioinsecticide (Netter et al., 2024 ). Similarly, researchers observed the insecticidal and synergistic potential of menthone, particularly against adults of the yellow fever mosquito ( Aedes aegypti Linnaeus, 1762 (Diptera: Culicidae)) and the housefly ( Musca domestica Linnaeus, 1758 (Diptera: Muscidae)) (Baker et al., 2023 ). In light of the results obtained in this study, it is evident that the M. piperita nanoemulsion not only exhibits satisfactory physicochemical stability but also demonstrates high biological efficacy against O. ilicis . The combination of major bioactive compounds and the use of a nanostructured formulation contribute to the toxic action observed, even at low concentrations and in the initial exposure periods (Heydari et al., 2020 ). These findings reinforce the potential of essential oil nanoemulsions as sustainable and effective alternatives for the management of coffee plant phytophagous mites, with the possibility of integration into biological control and integrated pest management (IPM) programs, offering reduced environmental impact and lower risk of resistance development. 5 Conclusion The findings of this study demonstrate that the M. piperita nanoemulsion combines desirable physicochemical stability with pronounced acaricidal efficacy against O. ilicis under laboratory conditions. Its rapid biological activity underscores its potential as a promising, environmentally sustainable tool for the management of the red coffee mite. Declarations Acknowledgments The authors would like to thank the Laboratory of Agricultural Entomology and Acarology (LEAA) and the Federal Institute of Espírito Santo (IFES) – Campus Itapina for their support in carrying out this study. Author Contributions Vanessa Racaneli Sian: Conceptualization, Investigation, Writing – Original Draft. Gustavo Pazolini Stein: Investigation, Writing – Review & Editing. Ana Beatriz Mamedes Piffer: Formal Analysis, Writing – Review & Editing. Plúcia Franciane Ataide Rodrigues: Methodology, Investigation. Anderson Mathias Holtz: Conceptualization, Supervision, Project Administration. Hildegado Seibert França: Methodology, Supervision, Resources. Funding This work was supported by the Instituto Federal do Espírito Santo (IFES), the Fundação de Amparo à Pesquisa e Inovação do Espírito Santo (FAPES), the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), and the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). Ethics statement Not applicable. Declaration of c onflict of Interest The authors declare that they have no conflict of interest. Data Availability Statement The data that support the findings of this study are all available in this article. References Abd-Elnabi AD, El-Sawy EA, Badawy ME. Plant oil nano-emulsions as a potential solution for pest control in sustainable agriculture. Neotrop Entomol . 54 (1):35 (2025). Doi:10.1007/s13744-024-01243-5 Adams, R. P., 2007. Identification of essential oil components by gas chromatography/mass spectrometry, 4th Edition. Allured Publ., Carol Stream, IL. Afiunizadeh M, Karimzadeh J, Imani S, Moharramipour S . Insecticidal and oviposition deterrent effects of five medicinal plant extracts on the diamondback moth. J Plant Diseases and Protec . 129 (4):805-817 (2022). Doi:10.1007/s41348-022-00592-w Agrofit – Phytosanitary Pesticide System [Internet]. 2025 [cited 2025 May 27]. Available from: https://agrofit.agricultura.gov.br/agrofit_cons/principal_agrofit_cons Albuquerque PM, Azevedo SG, Andrade CP, D’Ambros NCDS, Pérez MTM, Manzato L. Biotechnological applications of nanoencapsulated essential oils: A review. Polímeros . 14 (24):5495 (2022). Doi:10.3390/polym14245495 Annadurai KS, Chandrasekaran N, Velraja S, Hikku GS, Parvathi VD. Essential oil nanoemulsion: An emerging eco-friendly strategy towards mosquito control. Acta Trop . 257 :107290 (2024). Doi:10.1016/j.actatropica.2024.107290 Ayllón-Gutiérrez R, Díaz-Rubio L, Montaño-Soto M, Haro-Vázquez MDP, Córdova-Guerrero I. Applications of plant essential oils in pest control and their encapsulation for controlled release: a review. Agriculture . 14 (10):1766 (2024). Doi:10.3390/agriculture14101766 Baker OS, Norris EJ, Burgess ER. Insecticidal and synergistic potential of three monoterpenoids against Aedes aegypti (Diptera: Culicidae) and Musca domestica (Diptera: Muscidae). Molecules . 28 (7):3250 (2023). Doi:10.3390/molecules28073250 Carducci FC, de Batista Fonseca IC, dos Santos WG, Pereira CTM, Junior VM, Sera T, et al. Resistance to red mite in Coffea arabica genotype introgressed with Coffea racemosa genes. Aust J Crop Sci . 13 (5):683-6 (2019). Doi:10.21475/ajcs.19.13.05.p1254. Carvalho NBO, Lyra LF, Sakane KK. Aplicação da espectroscopia vibracional no infravermelho, acoplada a análises estatísticas multivariadas, para a análise dos efeitos do uso de herbicidas à base do Diuron em feijões carioca. Res Soc Dev . 12 (11):e136121143753 (2023). Doi:10.33448/rsd-v12i11.43753 Divekar PA, Narayana S, Divekar BA, Kumar R, Gadratagi BG, Ray A. et al. Plant secondary metabolites as defense tools against herbivores for sustainable crop protection . Int J Mol Sci . 23 (5):2690 (2022). Doi:10.3390/ijms23052690 Finetti L, Tiedemann L, Zhang X, Civolani S, Bernacchia G, Roeder T . Monoterpenes alter TAR1-driven physiology in Drosophila species. J Exp Biol . 224 (1): jeb232116 (2021). Doi:10.1242/jeb.232116 Gad HA, Ramadan GR, El-Bakry AM, El-Sabrout AM, Abdelgaleil SA . Monoterpenes: promising natural products for public health insect control – a review. Int J Trop Insect Sci. 42 (2):1059-75 (2022). Doi:10.1007/s42690-021-00692-4 Ghosh V, Mukherjee A, Chandrasekaran N. Optimization of process parameters for spontaneous emulsification-based nanoemulsion: evaluation of larvicidal activity against Culex quinquefasciatus . BioNanoScience . 4 (2):157-65 (2014). Doi:10.1007/s12668-014-0131-z Ghosh V, Ranjha R, Gupta AK. Polymeric encapsulation of anti-larval essential oil nanoemulsion for controlled release of bioactive compounds. Inorg Chem Commun . 150 :110507 (2023). Doi:10.1016/j.inoche.2023.110507 Heydari M, Amirjani A, Bagheri M, Sharifian I, Sabahi Q. Eco-friendly pesticide based on peppermint oil nanoemulsion: Preparation, physicochemical properties, and its aphicidal activity against cotton aphid. Environ Sci Poll Res 27 (6):6667-6679 (2020). Doi: 10.1007/s11356-019-07332-y Khan MH, Dar NA, Alie BA, Dar SA, Lone AA, Mir GH et al. Unraveling the variability of essential oil composition in different accessions of Bunium persicum collected from temperate micro-climates. Molecules . 28 (5):2404 (2023). Doi:10.3390/molecules28052404 Koroleva MY, Yurtov EV. Ostwald ripening in macro- and nanoemulsions. Russ Chem Rev . 90 (3):293 (2021). Doi:10.1070/RCR4962 Koundal R, Dolma SK, Chand G, Agnihotri VK, Reddy SE . Chemical composition and insecticidal properties of essential oils against diamondback moth ( Plutella xylostella L.). Toxin review . 41 (1):48-54 (2020). Doi:10.1080/15569543.2018.1536668 Lenth R. Emmeans: estimated marginal means, aka least-squares means. 2023 [Internet]. Available from: https://CRAN.R-project.org/package=emmeans Lima LA, Ferreira-Sá PS, Garcia Jr MD, Pereira VLP, Carvalho JCT, Rocha L. Nanoemulsions of the essential oil of Baccharis reticularia and its constituents as eco-friendly repellents against Tribolium castaneum . Ind Crops Prod. 162 :113282 (2021). Doi:10.1016/j.indcrop.2021.113282 Liu Z, Li QX, Song B. Pesticidal activity and mode of action of monoterpenes. J Agric Food Chem . 70 (15):4556-71 (2022). Doi:10.1021/acs.jafc.2c00635 Lopez L, Liburd OE. Injury to southern highbush blueberries by southern red mites and management using various miticides. Insects. 11 (4):233 (2020). Doi:10.3390/insects11040233 Marucci RC, Ruber SE, Pec M, Liburd, OE. Are predatory mites effective as biological control agents to suppress Oligonychus ilicis (Acari: Tetranychidae) in blueberry plantings?. J Economic Entom . 117 (3):834-842 (2024). Doi:10.1093/jee/toae086 McClements DJ. Nanoemulsions versus microemulsions: terminology, differences, and similarities. Soft Matter . 8 (6):1719-29 (2012). Doi:10.1039/C2SM06903B Modafferi A, Urbaneja A, Aure CM, Laudani F, Palmeri V, Giunti G, Pérez-Hedo M. Green pest control strategies: essential oil-based nano-emulsions for Delottococcus aberiae management. J Pest Sci 98 (3):1263-1275 (2025) Doi:10.1007/s10340-025-01914-1 Netter L, Silva YLL, Silva AB, Santos LV, Rocha, SBN. Atividade inseticida do óleo essencial de hortelã-pimenta sobre lagartas de Spodoptera frugiperda . Rev Ciênc Agríc . 22 (Esp):1-5 (2024). Doi:10.28998/rca.22 Oca-Ávalos JMM, Candal RJ, Herrera ML. Nanoemulsions: stability and physical properties . Curr Opin Food Sci. 16 :1-6 (2017). Doi:10.1016/j.cofs.2017.06.003 Ortiz-Zamora L, Bezerra DC, de Oliveira HNS, Duarte JL, Guisado-Bourzac F, Chil-Núñez I. Preparation of non-toxic nano-emulsions based on a Brazilian plant species through a low-energy concept. Ind Crops Prod . 158 :112989 (2020). Doi:10.1016/j.indcrop.2020.112989. Ozman SS, Sullivan G, Cakir S, Bas H, Saglam D, Doker I, et al. Phytoseiid mites: Trees, ecology and conservation. Diversity . 16 (9): 542 (2024). Doi:10.3390/d16090542 Paz Neto ADA, da Câmara CAG, Monteiro VB, de Moraes MM, de Melo JPR, Leal TTB . Bioactivity of essential oils from three species of Mentha L. against Plutella xylostella (Linnaeus, 1767)(Lepidoptera: Plutellidae). Entomologia Neotrop. 54 (1):101 (2025). Doi:10.1007/s13744-025-01317-y Piffer ABM, Holtz AM, Neto VB, de Paula Marchiori JJ, de Carvalho JR, Pretti IR, et al. Can soil conditioners of plant origin be used for pest control? Observ Econ Lat Am 21 (10): 15883–15900 (2023). Doi:10.55905/oelv21n10-075 R Core Team. R: a language and environment for statistical computing. 2023 [Internet]. Available from: http://www.R-project.org Reis PR, Alves EB, Sousa EO. Biologia do ácaro-vermelho do cafeeiro Oligonychus ilicis (McGregor, 1917). Ciência e Agrotecnologia , 21 : 260-266 (1997). Doi:10.5555/19981110067 Richardson LL, Adler LS, Leonard AS, Andicoechea J, Regan KH, Anthony WE, et al. Secondary metabolites in floral nectar reduce parasitic infections in bumblebees. Proc Biol Sci . 282 (1803):20142471 (2015). Doi:10.1098/rspb.2014.2471 Santos CAL, Moreira AMT, Teles BRDS, Kamdem JP, AlAsmari AF, Alasmari F. et al . Mentha arvensis oil exhibits repellent, acute toxic, and antioxidant activities in Nauphoeta cinerea . Sci Rep . 14 (1):21599 (2024). Doi:10.1038/s41598-024-72722-3 Souza LP, Zuim V, Stinguel P, Pinheiro PF, Zago HB. Toxicity of essential oil of Mentha piperita (Lamiaceae) and its monoterpenoid menthol against Tetranychus urticae Kogan 1836 (Acari: Tetranychidae). An Acad Bras Cienc. 94 (4):e20200427 (2022). Doi:10.1590/00013765202220200427 Souza RB, Guimarães JR. Effects of avermectins on the environment based on its toxicity to plants and soil invertebrates—a review. Water Air Soil Pollut . 233 (7):259 (2022). Doi:10.1007/s11270-022-05744-0 Stringaro A, Colone M, Angiolella L. Antioxidant, antifungal, antibiofilm and cytotoxic activities of Mentha spp essential oils. Medicines . 5 (4):112 (2018). Doi:0.3390/medicamentos5040112 Wu Z, Jin C, Chen Y, Yang S, Yang X, Zhang D, Xie Y. Essential oils of Mentha spp . as a potential toxic fumigant with acetylcholinesterase inhibition in Reticulitermes dabieshanensis . Plants. 12 (23):4034 (2023). Doi:10.3390/plants12234034 Zhang W, Jiang H, Rhim JW, Cao J, Jiang W. Effective strategies of sustained release and retention enhancement of essential oils in active food packaging films/coatings . Food Chem. 367: 130671 (2022). Doi:10.1016/j.foodchem.2021.130671 Table Table 2 is available in the Supplementary Files section. Additional Declarations No competing interests reported. Supplementary Files Table2.docx Resumogrficoingles.pdf Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 20 Apr, 2026 Reviews received at journal 20 Apr, 2026 Reviews received at journal 16 Apr, 2026 Reviewers agreed at journal 15 Apr, 2026 Reviews received at journal 11 Apr, 2026 Reviewers agreed at journal 11 Apr, 2026 Reviewers agreed at journal 10 Apr, 2026 Reviewers agreed at journal 09 Apr, 2026 Reviewers agreed at journal 08 Apr, 2026 Reviewers invited by journal 08 Apr, 2026 Editor assigned by journal 08 Apr, 2026 Submission checks completed at journal 08 Apr, 2026 First submitted to journal 29 Mar, 2026 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-9260778","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":623438392,"identity":"315ac741-a224-4ba9-95a3-301224adebda","order_by":0,"name":"Vanessa Racaneli Sian","email":"data:image/png;base64,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","orcid":"","institution":"Instituto Federal do Espírito Santo","correspondingAuthor":true,"prefix":"","firstName":"Vanessa","middleName":"Racaneli","lastName":"Sian","suffix":""},{"id":623438393,"identity":"6728ebcd-668a-46aa-932a-ecd7df148c67","order_by":1,"name":"Gustavo Pazolini Stein","email":"","orcid":"","institution":"Instituto Federal do Espírito Santo","correspondingAuthor":false,"prefix":"","firstName":"Gustavo","middleName":"Pazolini","lastName":"Stein","suffix":""},{"id":623438394,"identity":"57f38da9-88e5-4015-8d13-c00b1e10617c","order_by":2,"name":"Ana Beatriz Mamedes Piffer","email":"","orcid":"","institution":"Universidade Federal de Viçosa","correspondingAuthor":false,"prefix":"","firstName":"Ana","middleName":"Beatriz Mamedes","lastName":"Piffer","suffix":""},{"id":623438395,"identity":"90718210-99a2-4f2f-9ca7-b46393482b6c","order_by":3,"name":"Plúcia Franciane Ataide Rodrigues","email":"","orcid":"","institution":"Instituto Federal do Espírito Santo","correspondingAuthor":false,"prefix":"","firstName":"Plúcia","middleName":"Franciane Ataide","lastName":"Rodrigues","suffix":""},{"id":623438402,"identity":"51e7587d-53c3-4114-9b7b-f30d17f440c0","order_by":4,"name":"Anderson Mathias Holtz","email":"","orcid":"","institution":"Instituto Federal do Espírito Santo","correspondingAuthor":false,"prefix":"","firstName":"Anderson","middleName":"Mathias","lastName":"Holtz","suffix":""},{"id":623438403,"identity":"8cbded78-5b50-45f8-895a-9d1d48fc675d","order_by":5,"name":"Hildegado Seibert França","email":"","orcid":"","institution":"Instituto Federal do Espírito Santo","correspondingAuthor":false,"prefix":"","firstName":"Hildegado","middleName":"Seibert","lastName":"França","suffix":""}],"badges":[],"createdAt":"2026-03-29 19:53:47","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-9260778/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-9260778/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":107085499,"identity":"3504ee5f-719e-4705-a0cc-2fb7e5e14d5c","added_by":"auto","created_at":"2026-04-16 15:00:11","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":136517,"visible":true,"origin":"","legend":"\u003cp\u003eGeographic location of IFES – Itapina Campus.\u003c/p\u003e","description":"","filename":"Figure1.png","url":"https://assets-eu.researchsquare.com/files/rs-9260778/v1/2e09ea86025374e5d57bd261.png"},{"id":107481451,"identity":"e3b29ae2-c3cc-46a4-9140-994c98acec48","added_by":"auto","created_at":"2026-04-22 02:18:02","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":648263,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eM. piperita\u003c/em\u003e essential oil nanoemulsions, with respective concentrations and storage times after production.\u003c/p\u003e","description":"","filename":"Figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-9260778/v1/615a392aa4f7b86ff7705357.png"},{"id":107705059,"identity":"a078769e-57fd-4a84-9a4c-586a1efec781","added_by":"auto","created_at":"2026-04-24 09:07:18","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":60322,"visible":true,"origin":"","legend":"\u003cp\u003eChromatogram of Mentha piperita obtained by gas chromatography coupled with mass spectrometry (GC-MS).\u003c/p\u003e","description":"","filename":"Figure3.png","url":"https://assets-eu.researchsquare.com/files/rs-9260778/v1/d01b46bd990971d1a125db6e.png"},{"id":107085504,"identity":"37993a56-936b-4a92-b240-5b9713f58e6a","added_by":"auto","created_at":"2026-04-16 15:00:11","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":672322,"visible":true,"origin":"","legend":"\u003cp\u003eStability test, particle size, and zeta potential of the most stable concentrations of \u003cem\u003eMentha piperita\u003c/em\u003e nanoemulsion.\u003c/p\u003e","description":"","filename":"Figure4.png","url":"https://assets-eu.researchsquare.com/files/rs-9260778/v1/ceaa1ccb436d9b50a5aa03ee.png"},{"id":107480838,"identity":"bb5e5920-e4c1-46f1-b447-0ed5c633e49b","added_by":"auto","created_at":"2026-04-22 02:13:47","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":7554,"visible":true,"origin":"","legend":"\u003cp\u003eOverlap of FT-IR spectra of \u003cem\u003eMentha piperita\u003c/em\u003e essential oil, surfactant, and nanoemulsion.\u003c/p\u003e","description":"","filename":"Figure5.png","url":"https://assets-eu.researchsquare.com/files/rs-9260778/v1/d60724556450e3c55db01585.png"},{"id":107085506,"identity":"e173e94a-9bea-462b-b4bc-eea4617bc22a","added_by":"auto","created_at":"2026-04-16 15:00:11","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":6417,"visible":true,"origin":"","legend":"\u003cp\u003eMortality dynamics of \u003cem\u003eOligonychus ilicis\u003c/em\u003e under different concentrations of \u003cem\u003eMentha piperita\u003c/em\u003e essential oil nanoemulsion over 12, 24, 36, 48, 60, and 72 hours. Temp.: 25 ± 1 °C, RH 70 ± 10%, and 12 h photoperiod.\u003c/p\u003e","description":"","filename":"Figure6.png","url":"https://assets-eu.researchsquare.com/files/rs-9260778/v1/5acc099b3032bc05fcef893e.png"},{"id":107708713,"identity":"1a1b8913-081b-46c5-92a2-6ffbe666be36","added_by":"auto","created_at":"2026-04-24 09:31:44","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1897158,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9260778/v1/20b8aabb-09fa-4e18-bfe4-ce9a86384ecc.pdf"},{"id":107481544,"identity":"64f37a2d-5f3e-4fcb-b871-d6beaef13a93","added_by":"auto","created_at":"2026-04-22 02:18:56","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":25168,"visible":true,"origin":"","legend":"","description":"","filename":"Table2.docx","url":"https://assets-eu.researchsquare.com/files/rs-9260778/v1/7d58b3a9f3b1ff0461aef524.docx"},{"id":107085501,"identity":"3dd05f5b-c440-40f9-ba33-603e59891ce0","added_by":"auto","created_at":"2026-04-16 15:00:11","extension":"pdf","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":3144305,"visible":true,"origin":"","legend":"","description":"","filename":"Resumogrficoingles.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9260778/v1/265e875aad6d13ec5dd0d4d8.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Nanoemulsion based on essential oil of Mentha piperita in the sustainable management of Oligonychus ilicis McGregor, 1917 (Acari: Tetranychidae)","fulltext":[{"header":"1 Introduction","content":"\u003cp\u003eThe red mite, \u003cem\u003eOligonychus ilicis\u003c/em\u003e McGregor, 1917 (Acari: Tetranychidae), is a phytophagous arthropod of major economic importance and is described as one of the key pests of Conilon coffee (\u003cem\u003eCoffea canephora\u003c/em\u003e Pierre \u0026amp; Froehn) (Piffer et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). This species preferentially colonizes the adaxial surface of leaves, where it pierces epidermal cells to feed, extracting part of the cellular contents and imparting a characteristic bronzed appearance to the foliage. In association with its presence, a delicate web produced by the mites themselves can accumulate dust and debris, giving leaves a dirty appearance (Ozman et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Prolonged dry periods with water deficit are favorable for mite outbreaks and may lead to severe defoliation. In young plantations, the damage caused by this pest reduces photosynthetic capacity, compromises vegetative growth and vigor, and can directly affect crop survival (Lopez and Liburd, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Marucci et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAccording to the Ministry of Agriculture, Livestock and Supply (MAPA), there are approximately 70 products registered for the control of the red coffee mite, of which only two are classified as environmentally safe (Agrofit, 2025). Most of these products contain avermectin-based active ingredients, which not only cause mortality and reduced reproduction in soil invertebrates but may also induce phytotoxic effects in plants (Souza and Guimar\u0026atilde;es, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Moreover, synthetic chemicals have adverse effects on non-target organisms and aquatic resources (Carducci et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). To date, there are no officially registered biological products for the management of \u003cem\u003eO. ilicis\u003c/em\u003e, highlighting the urgent need for research and the development of sustainable and environmentally safer alternatives (Agrofit, 2025).\u003c/p\u003e \u003cp\u003eEssential oils (EOs) are metabolites produced through the secondary metabolism of plants as a defense mechanism against arthropods. Their bioactivity is associated with phytochemical classes such as terpenoids, alkaloids, flavonoids, steroids, saponins, and tannins, many of which are naturally toxic to pests (Richardson et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Divekar et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Among the plant species with reported acaricidal properties, peppermint (\u003cem\u003eMentha piperita\u003c/em\u003e) is noteworthy (Souza et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Peppermint is a perennial aromatic herb containing important compounds such as menthol, menthone, limonene, isomenthone, menthyl acetate, carvone, β-pinene, and 1,8-cineole, all of which have demonstrated activity against pest organisms (Richardson et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Stringaro et al., \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Divekar et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). However, under environmental conditions, EOs are chemically unstable and undergo rapid degradation (Zhang et al., \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Albuquerque et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eNanoemulsions have attracted increasing attention in agriculture due to their ability to stabilize lipophilic active ingredients, provide controlled release and prolonged bioactivity, and enhance the chemical stability of formulations by preventing phase separation (Ghosh et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2014\u003c/span\u003e, Divekar et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Furthermore, these formulations represent a promising alternative to synthetic chemicals, as they tend to exert lower environmental impact (Annadurai et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Modafferi et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). In this context, the objective of this study was to characterize the chemical composition of \u003cem\u003eM. piperita\u003c/em\u003e essential oil, formulate a peppermint oil-based nanoemulsion, and evaluate its acaricidal effect against \u003cem\u003eO. ilicis\u003c/em\u003e under laboratory conditions.\u003c/p\u003e"},{"header":"2 Materials and Methods","content":"\u003cp\u003eThe experiments were conducted at the Laboratory of Entomology and Agricultural Acarology of the Federal Institute of Education, Science and Technology of Esp\u0026iacute;rito Santo \u0026ndash; Itapina Campus (IFES \u0026ndash; Itapina Campus), located in the municipality of Colatina, at the geographic coordinates 19\u0026deg;29'52.7\"S and 40\u0026deg;45'38.5\"W (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The bioassays were carried out in climate-controlled chambers at 25\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C, 70\u0026thinsp;\u0026plusmn;\u0026thinsp;10% relative humidity, and a 12 h photoperiod.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Essential oil acquisition and chemical composition analysis\u003c/h2\u003e \u003cp\u003eThe essential oil was commercially obtained from Ferquima Ind\u0026uacute;stria e Com\u0026eacute;rcio Ltda (Batch 218). The chemical characterization of the essential oil was performed by gas chromatography coupled with mass spectrometry (GC-MS), using an Agilent 7890B gas chromatograph coupled to a 5977A MSD mass selective detector, operating with electron impact ionization at 70 eV. Compound separation was carried out using an HP-5 capillary column (30 m \u0026times; 250 \u0026micro;m \u0026times; 0.25 \u0026micro;m). The injector and detector temperatures were set at 290\u0026deg;C and 310\u0026deg;C, respectively.\u003c/p\u003e \u003cp\u003eThe oven temperature program consisted of an initial temperature of 40\u0026deg;C, followed by a heating ramp of 5\u0026deg;C/min up to 280\u0026deg;C, and a second ramp of 15\u0026deg;C/min up to a final temperature of 310\u0026deg;C. For the calculation of Kovats retention indices (KI), a standard mixture of n-alkanes ranging from C10 to C40 was used. Compound identification was based on comparison of the obtained mass spectra with the NIST library data, as well as correlation of retention indices and spectral patterns with reference data from the literature (Adams, 2017).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Formulation and physicochemical characterization of the \u003cem\u003eMentha piperita\u003c/em\u003e nanoemulsion\u003c/h2\u003e \u003cp\u003eThe peppermint essential oil-based nanoemulsion was obtained using the low-energy emulsification method, with adaptations (McClements, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). Initially, 24 combinations varying in hydrophilic-lipophilic balance (HLB) values and surfactant ratios were tested to identify the most stable formulation. The oil phase (essential oil\u0026thinsp;+\u0026thinsp;surfactant) was homogenized by vortexing, after which the aqueous phase (distilled water) was slowly added under continuous stirring until reaching a final mass of 1 g (50 mg/mL). The most stable \u003cem\u003eM. piperita\u003c/em\u003e nanoemulsion was achieved at HLB 15, using 5% essential oil, 20% surfactants (Polysorbate 20 and Sorbitan monooleate), and 75% water.\u003c/p\u003e \u003cp\u003eSubsequently, the formulation was diluted to concentrations of 2.5%, 1%, 0.5%, and 0.2% essential oil. Nanoemulsion stability was monitored on days 0, 17, and 34 after preparation through visual assessment of macroscopic characteristics, following certain criteria (Ortiz-Zamora et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Lima et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The presence of a continuous phase and the Tyndall effect was observed by a bluish coloration under incident light, indicating the formation of nanostructures in the formulations.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe dilutions that remained macroscopically stable (1, 0.5, and 0.2%) were subjected to hydrodynamic diameter, polydispersity index (PDI), and zeta potential analyses. FT-IR analyses were performed using a Spectrum Two spectrometer equipped with a diamond ATR (Attenuated Total Reflection) accessory. The PerkinElmer Spectrum software was used for spectral processing. Spectra were acquired with 32 scans per spectrum of the surfactants, in triplicate, within the mid-infrared range of 4000\u0026ndash;400 cm⁻\u0026sup1;, with a spectral resolution of 8 cm⁻\u0026sup1;, adapted (Carvalho et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eColloidal characterization was carried out by dynamic light scattering (DLS) using a Litesizer 500\u0026reg; (Anton Paar) to determine the average hydrodynamic diameter, polydispersity index (PDI), and zeta potential, with a 1:400 (v/v) dilution in distilled water. Measurements were taken on days 1, 17, and 34 after preparation at concentrations of 1%, 0.5%, and 0.2%, with up to 1000 runs per sample.\u003c/p\u003e \u003cp\u003eBased on stability analyses and the maintenance of physicochemical parameters observed at concentrations of 0.5% and 0.2%, lower dilutions were selected for the bioassays against mites. Thus, concentrations of 0.025%, 0.040%, 0.060%, 0.10%, 0.15%, and 0.25% were used in the biological tests, obtained from progressive dilution of the previously stabilized formulation.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Rearing of \u003cem\u003eOligonychus ilicis\u003c/em\u003e\u003c/h2\u003e \u003cp\u003eMites were collected from a coffee plantation at IFES \u0026ndash; Itapina Campus, where no chemical products had been applied, and subsequently reared under laboratory conditions following an adaptation of the technique (Reis et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e1997\u003c/span\u003e). In the same plantation, leaves were collected, washed in distilled water, and disinfected in a sodium dichloroisocyanurate solution (1:10). Petri dishes (14.0 \u0026times; 1.5 cm) were prepared as rearing arenas, with moistened cotton placed at the bottom and around the coffee leaf to maintain leaf turgor and prevent mite escape. The arenas were maintained in climate-controlled B.O.D. chambers at 25\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C, 70\u0026thinsp;\u0026plusmn;\u0026thinsp;10% relative humidity, and a 12 h photoperiod, with weekly maintenance to ensure colony preservation under laboratory conditions.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Laboratory bioassays with essential oil-based nanoemulsions against \u003cem\u003eOligonychus ilicis\u003c/em\u003e\u003c/h2\u003e \u003cp\u003eFor the bioassays, \u003cem\u003eO. ilicis\u003c/em\u003e females were standardized to obtain adult individuals of uniform age. Arenas were assembled in Petri dishes (10.0 \u0026times; 1.2 cm) containing coffee leaf discs (4 cm in diameter), with moistened cotton placed around the discs to maintain leaf turgor. Ten females were carefully transferred to each dish using a fine-bristle brush.\u003c/p\u003e \u003cp\u003eThe nanoemulsion was tested at concentrations of 0.025, 0.040, 0.060, 0.10, 0.15, and 0.25% (mL mL⁻\u0026sup1;), obtained from the base formulation. Each treatment consisted of 10 replicates. The control treatment consisted of distilled water with Tween\u0026reg; 80 (0.05%) as a surfactant. Application of the solutions to the mites was performed using an airbrush calibrated at a constant pressure of 15 psi, delivering 1 mL of solution per replicate.\u003c/p\u003e \u003cp\u003eMite mortality was assessed at 12, 24, 36, 48, 60, and 72 hours after spraying. Individuals were considered dead when they failed to move after gentle stimulation with the brush bristles.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.5 Statistical analysis\u003c/h2\u003e \u003cp\u003eData were subjected to Probit regression analysis, and the resulting equations were used to estimate LC₅₀ and LC₉₀ values. Survival data were analyzed using a Cox proportional hazards model, and treatment contrasts were evaluated with the emmeans function of the corresponding R package (Lenth, 2023). All statistical analyses were performed using R software, version 4.3.2 (R Core Team, 2023).\u003c/p\u003e \u003c/div\u003e"},{"header":"3 Results","content":"\u003cp\u003e\u003cstrong\u003e3.1 Chemical composition of peppermint oil by gas chromatography\u0026ndash;mass spectrometry\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTwelve compounds were identified, accounting for 99.4% of the total peak area (Table 1). Among them, the major constituents were isomenthol and isomenthone, which represented 67.4% of the oil (Fig. 3). The presence of monoterpene ketones, monoterpene alcohols, monoterpene ethers, and hydrocarbons was detected. When correlating the absorption bands observed in the FT-IR spectra with the compounds identified in the essential oil of \u003cem\u003eM. piperita\u003c/em\u003e (Table 2), a large number of bands below 1500 cm⁻\u0026sup1; were observed, which is associated with the diversity of functional groups among the substances identified by GC-MS.\u003c/p\u003e\n\u003cp\u003eIn addition to the bands related to alkane and olefin vibrations, sharp and intense peaks were observed, corresponding to the C=O stretching vibration at 1750\u0026ndash;1700 cm⁻\u0026sup1;, the C\u0026ndash;O stretching vibration at 1050 cm⁻\u0026sup1;, and the asymmetric C\u0026ndash;O\u0026ndash;C stretching vibration at 1280\u0026ndash;1230 cm⁻\u0026sup1;.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1\u0026nbsp;\u003c/strong\u003eMajor chemical constituents of \u003cem\u003eMentha piperita\u003c/em\u003e essential oil identified by GC-MS analysis.\u003c/p\u003e\n\u003cdiv align=\"\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"301\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 53.9735%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCompound name\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46.0265%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRelative area (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 53.9735%;\"\u003e\n \u003cp\u003eMenthol (isomer)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46.0265%;\"\u003e\n \u003cp\u003e42,079\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 53.9735%;\"\u003e\n \u003cp\u003eMenthone\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46.0265%;\"\u003e\n \u003cp\u003e25,323\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 53.9735%;\"\u003e\n \u003cp\u003eMenthol (neoisomer)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46.0265%;\"\u003e\n \u003cp\u003e8,759\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 53.9735%;\"\u003e\n \u003cp\u003eMenthyl acetate\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46.0265%;\"\u003e\n \u003cp\u003e7,616\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 53.9735%;\"\u003e\n \u003cp\u003e1,8-Cineole\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46.0265%;\"\u003e\n \u003cp\u003e6,49\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 53.9735%;\"\u003e\n \u003cp\u003eLimonene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46.0265%;\"\u003e\n \u003cp\u003e2,544\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 53.9735%;\"\u003e\n \u003cp\u003eCaryophyllene oxide\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46.0265%;\"\u003e\n \u003cp\u003e1,683\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 53.9735%;\"\u003e\n \u003cp\u003e\u0026beta;-Pinene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46.0265%;\"\u003e\n \u003cp\u003e1,258\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 53.9735%;\"\u003e\n \u003cp\u003eCaryophyllene (E)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46.0265%;\"\u003e\n \u003cp\u003e1,159\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 53.9735%;\"\u003e\n \u003cp\u003ePulegone\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46.0265%;\"\u003e\n \u003cp\u003e1,091\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 53.9735%;\"\u003e\n \u003cp\u003e\u0026alpha;-Pinene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46.0265%;\"\u003e\n \u003cp\u003e0,996\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 53.9735%;\"\u003e\n \u003cp\u003ePiperitone\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46.0265%;\"\u003e\n \u003cp\u003e0,433\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003e3.2 Macroscopic stability, hydrodynamic size, and zeta potential analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe peppermint essential oil-based nanoemulsion at different concentrations was characterized in terms of droplet size, hydrodynamic diameter, polydispersity index (PDI), zeta potential, and particle size distribution peaks over time. The use of the low-energy input method enabled the development of highly stable nanoemulsions, with droplet sizes in the nanometric range, low PDI values, and good overall stability (Fig. 4).\u003c/p\u003e\n\u003cp\u003eInitially, all concentrations exhibited stability. The 1% dilution, although displaying a whitish appearance, did not undergo phase separation and retained nanometric droplet size; however, it did not remain stable over time (Fig. 4). In contrast, the 0.2% and 0.5% dilutions remained stable until the final day of analysis, without significant differences in hydrodynamic diameter between them. Furthermore, the zeta potential and polydispersity index values demonstrated stability (Fig. 4). The dilutions exhibited hydrodynamic diameters of 59.25 nm and 58.64 nm, respectively.\u003c/p\u003e\n\u003cp\u003eBased on these data, and considering the visual homogeneity observed in the samples, it can be inferred that even lower dilutions, such as those used in the bioassays (0.025% to 0.25%), exhibit equivalent physicochemical behavior, maintaining the integrity of the nanostructured system. The stability observed in the 0.5% and 0.2% samples, together with the absence of flocculation or phase separation and the preservation of colloidal properties, supports this inference, despite the instrumental limitation in directly measuring such lower concentrations.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.5 Toxicity effect of the nanoemulsion based on \u003cem\u003eMentha piperita\u003c/em\u003e essential oil\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAn acaricidal effect of the peppermint essential oil-based nanoemulsion was verified, with adult mortality of \u003cem\u003eO. ilicis\u003c/em\u003e increasing in a dose-dependent manner. The lethal concentrations for 50% and 90% of the population were estimated at 0.0257 and 0.0424 (mL\u0026middot;mL⁻\u0026sup1;), respectively (Fig.5). In the survival analysis of \u003cem\u003eO. ilicis\u003c/em\u003e, the highest mortality rates occurred within 12 hours after direct application on the individuals, reaching up to 100% mortality at concentrations of 0.15% and 0.25% (Fig. 6).\u003c/p\u003e"},{"header":"4 Discussion","content":"\u003cp\u003eNanoemulsions, due to their nanometric-scale droplets, are structurally more susceptible to destabilization by Ostwald ripening (Koroleva and Yurtov, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). However, this same reduced size favors their kinetic stability by minimizing phenomena such as flocculation, coalescence, and gravitational separation, particularly in well-balanced formulations (Oca-\u0026Aacute;valos et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). With lower hydrodynamic diameter and polydispersity index values, the results obtained in this study are consistent with the standards established by some researchers, indicating the development of a stable and promising system (McClements, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). Such stability favors the controlled and prolonged release of the phytochemical compounds present in the essential oil, contributing to the lethal effects observed in the bioassays (Ghosh et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Ayll\u0026oacute;n-Guti\u0026eacute;rrez et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe toxicity of essential oils may be directly related to the diversity and concentration of their active constituents, which vary according to agronomic, edaphoclimatic, and methodological factors, such as soil type, extraction method, and plant phenological stage, among others (Khan et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Abd-Elnabi et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). The chemical analysis of \u003cem\u003eM. piperita\u003c/em\u003e essential oil revealed the presence of compounds belonging to three main chemical groups: hydrocarbon monoterpenes, oxygenated monoterpenes, and oxygenated sesquiterpenes. Among these, oxygenated monoterpenes stand out, particularly menthol and menthone, which emerged as the major constituents. These compounds have been extensively studied due to their recognized insecticidal and repellent properties (Baker et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eMenthol is capable of inhibiting the enzyme acetylcholinesterase (AChE), leading to the accumulation of acetylcholine at synapses, resulting in nervous hyperexcitation, paralysis, and insect death (Finetti et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Liu et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Menthone, on the other hand, although also presenting this effect, has its activity more associated with the modulation of ion channels, especially sodium channels and GABA receptors, altering nerve impulse conduction and causing motor incoordination (Gad et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Souza et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Wu et al., \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Santos et al., \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eResearch has shown that the essential oils of \u003cem\u003eM. piperita\u003c/em\u003e exhibit residual contact toxicity and exert a repellent and food deterrent effect on third-instar larvae of the diamondback moth (\u003cem\u003ePlutella xylostella\u003c/em\u003e L., Lepidoptera: Plutellidae)(Koundal \u003cem\u003eet al.\u003c/em\u003e, 2024; Paz \u003cem\u003eet al.\u003c/em\u003e, 2025). Additionally, the methanolic extract of \u003cem\u003eM. piperita\u003c/em\u003e is demonstrated as effective as a larvicide and oviposition inhibitor for this pest (Afiunizadeh et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Koundal \u003cem\u003eet al.\u003c/em\u003e, 2024; Paz \u003cem\u003eet al.\u003c/em\u003e, 2025). Also, researchers reported effective control of the fall armyworm (\u003cem\u003eSpodoptera frugiperda\u003c/em\u003e (Lepidoptera: Noctuidae)) using peppermint oil, reinforcing its potential as a broad-spectrum bioinsecticide (Netter et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Similarly, researchers observed the insecticidal and synergistic potential of menthone, particularly against adults of the yellow fever mosquito (\u003cem\u003eAedes aegypti\u003c/em\u003e Linnaeus, 1762 (Diptera: Culicidae)) and the housefly (\u003cem\u003eMusca domestica\u003c/em\u003e Linnaeus, 1758 (Diptera: Muscidae)) (Baker et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn light of the results obtained in this study, it is evident that the \u003cem\u003eM. piperita\u003c/em\u003e nanoemulsion not only exhibits satisfactory physicochemical stability but also demonstrates high biological efficacy against \u003cem\u003eO. ilicis\u003c/em\u003e. The combination of major bioactive compounds and the use of a nanostructured formulation contribute to the toxic action observed, even at low concentrations and in the initial exposure periods (Heydari et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). These findings reinforce the potential of essential oil nanoemulsions as sustainable and effective alternatives for the management of coffee plant phytophagous mites, with the possibility of integration into biological control and integrated pest management (IPM) programs, offering reduced environmental impact and lower risk of resistance development.\u003c/p\u003e"},{"header":"5 Conclusion","content":"\u003cp\u003eThe findings of this study demonstrate that the \u003cem\u003eM. piperita\u003c/em\u003e nanoemulsion combines desirable physicochemical stability with pronounced acaricidal efficacy against \u003cem\u003eO. ilicis\u003c/em\u003e under laboratory conditions. Its rapid biological activity underscores its potential as a promising, environmentally sustainable tool for the management of the red coffee mite.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors would like to thank the Laboratory of Agricultural Entomology and Acarology (LEAA) and the Federal Institute of Esp\u0026iacute;rito Santo (IFES) \u0026ndash; Campus Itapina for their support in carrying out this study.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eVanessa Racaneli Sian: Conceptualization, Investigation, Writing \u0026ndash; Original Draft. Gustavo Pazolini Stein: Investigation, Writing \u0026ndash; Review \u0026amp; Editing. Ana Beatriz Mamedes Piffer: Formal Analysis, Writing \u0026ndash; Review \u0026amp; Editing. Pl\u0026uacute;cia Franciane Ataide Rodrigues: Methodology, Investigation. Anderson Mathias Holtz: Conceptualization, Supervision, Project Administration. Hildegado Seibert Fran\u0026ccedil;a: Methodology, Supervision, Resources.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by the Instituto Federal do Esp\u0026iacute;rito Santo (IFES), the Funda\u0026ccedil;\u0026atilde;o de Amparo \u0026agrave; Pesquisa e Inova\u0026ccedil;\u0026atilde;o do Esp\u0026iacute;rito Santo (FAPES), the Coordena\u0026ccedil;\u0026atilde;o de Aperfei\u0026ccedil;oamento de Pessoal de N\u0026iacute;vel Superior (CAPES), and the Conselho Nacional de Desenvolvimento Cient\u0026iacute;fico e Tecnol\u0026oacute;gico (CNPq).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDeclaration of c\u003c/strong\u003e\u003cstrong\u003eonflict of Interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no conflict of interest.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability Statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data that support the findings of this study are all available in this article.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAbd-Elnabi AD, El-Sawy EA, Badawy ME. Plant oil nano-emulsions as a potential solution for pest control in sustainable agriculture. \u003cem\u003eNeotrop Entomol\u003c/em\u003e.\u003cstrong\u003e54\u003c/strong\u003e(1):35 (2025). Doi:10.1007/s13744-024-01243-5\u003c/li\u003e\n\u003cli\u003eAdams, R. P., 2007. Identification of essential oil components by gas chromatography/mass spectrometry, 4th Edition. Allured Publ., Carol Stream, IL.\u003c/li\u003e\n\u003cli\u003eAfiunizadeh M, Karimzadeh J, Imani S, Moharramipour S\u003cem\u003e.\u003c/em\u003e Insecticidal and oviposition deterrent effects of five medicinal plant extracts on the diamondback moth. \u003cem\u003eJ Plant Diseases and Protec\u003c/em\u003e.\u003cstrong\u003e129\u003c/strong\u003e(4):805-817 (2022). Doi:10.1007/s41348-022-00592-w\u003c/li\u003e\n\u003cli\u003eAgrofit \u0026ndash; Phytosanitary Pesticide System [Internet]. 2025 [cited 2025 May 27]. Available from: https://agrofit.agricultura.gov.br/agrofit_cons/principal_agrofit_cons\u003c/li\u003e\n\u003cli\u003eAlbuquerque PM, Azevedo SG, Andrade CP, D\u0026rsquo;Ambros NCDS, P\u0026eacute;rez MTM, Manzato L. Biotechnological applications of nanoencapsulated essential oils: A review. \u003cem\u003ePol\u0026iacute;meros\u003c/em\u003e.\u003cstrong\u003e14\u003c/strong\u003e(24):5495 (2022). Doi:10.3390/polym14245495\u003c/li\u003e\n\u003cli\u003eAnnadurai KS, Chandrasekaran N, Velraja S, Hikku GS, Parvathi VD. Essential oil nanoemulsion: An emerging eco-friendly strategy towards mosquito control. \u003cem\u003eActa Trop\u003c/em\u003e.\u003cstrong\u003e257\u003c/strong\u003e:107290 (2024). Doi:10.1016/j.actatropica.2024.107290\u003c/li\u003e\n\u003cli\u003eAyll\u0026oacute;n-Guti\u0026eacute;rrez R, D\u0026iacute;az-Rubio L, Monta\u0026ntilde;o-Soto M, Haro-V\u0026aacute;zquez MDP, C\u0026oacute;rdova-Guerrero I. Applications of plant essential oils in pest control and their encapsulation for controlled release: a review. \u003cem\u003eAgriculture\u003c/em\u003e.\u003cstrong\u003e14\u003c/strong\u003e(10):1766 (2024). Doi:10.3390/agriculture14101766\u003c/li\u003e\n\u003cli\u003eBaker OS, Norris EJ, Burgess ER. Insecticidal and synergistic potential of three monoterpenoids against \u003cem\u003eAedes aegypti\u003c/em\u003e (Diptera: Culicidae) and \u003cem\u003eMusca \u003c/em\u003edomestica (Diptera: Muscidae). \u003cem\u003eMolecules\u003c/em\u003e.\u003cstrong\u003e28\u003c/strong\u003e(7):3250 (2023). Doi:10.3390/molecules28073250\u003c/li\u003e\n\u003cli\u003eCarducci FC, de Batista Fonseca IC, dos Santos WG, Pereira CTM, Junior VM, Sera T, \u003cem\u003eet al.\u003c/em\u003e Resistance to red mite in \u003cem\u003eCoffea arabica\u003c/em\u003e genotype introgressed with \u003cem\u003eCoffea racemosa\u003c/em\u003e genes.\u003cem\u003eAust J Crop Sci\u003c/em\u003e.\u003cstrong\u003e13\u003c/strong\u003e(5):683-6 (2019). Doi:10.21475/ajcs.19.13.05.p1254. \u003c/li\u003e\n\u003cli\u003eCarvalho NBO, Lyra LF, Sakane KK. Aplica\u0026ccedil;\u0026atilde;o da espectroscopia vibracional no infravermelho, acoplada a an\u0026aacute;lises estat\u0026iacute;sticas multivariadas, para a an\u0026aacute;lise dos efeitos do uso de herbicidas \u0026agrave; base do Diuron em feij\u0026otilde;es carioca. \u003cem\u003eRes Soc Dev\u003c/em\u003e.\u003cstrong\u003e12\u003c/strong\u003e(11):e136121143753 (2023). Doi:10.33448/rsd-v12i11.43753\u003c/li\u003e\n\u003cli\u003eDivekar PA, Narayana S, Divekar BA, Kumar R, Gadratagi BG, Ray A. \u003cem\u003eet al. \u003c/em\u003ePlant secondary metabolites as defense tools against herbivores for sustainable crop protection\u003cem\u003e. Int J Mol Sci\u003c/em\u003e.\u003cstrong\u003e23\u003c/strong\u003e(5):2690 (2022). Doi:10.3390/ijms23052690 \u003c/li\u003e\n\u003cli\u003eFinetti L, Tiedemann L, Zhang X, Civolani S, Bernacchia G, Roeder T\u003cem\u003e.\u003c/em\u003e Monoterpenes alter TAR1-driven physiology in \u003cem\u003eDrosophila\u003c/em\u003e species. \u003cem\u003eJ Exp Biol\u003c/em\u003e.\u003cstrong\u003e224\u003c/strong\u003e(1): jeb232116 (2021). Doi:10.1242/jeb.232116\u003c/li\u003e\n\u003cli\u003eGad HA, Ramadan GR, El-Bakry AM, El-Sabrout AM, Abdelgaleil SA\u003cem\u003e.\u003c/em\u003e Monoterpenes: promising natural products for public health insect control \u0026ndash; a review. \u003cem\u003eInt J Trop Insect Sci.\u003c/em\u003e\u003cstrong\u003e42\u003c/strong\u003e(2):1059-75 (2022). Doi:10.1007/s42690-021-00692-4\u003c/li\u003e\n\u003cli\u003eGhosh V, Mukherjee A, Chandrasekaran N. Optimization of process parameters for spontaneous emulsification-based nanoemulsion: evaluation of larvicidal activity against \u003cem\u003eCulex quinquefasciatus\u003c/em\u003e.\u003cem\u003eBioNanoScience\u003c/em\u003e. \u003cstrong\u003e4\u003c/strong\u003e(2):157-65 (2014). Doi:10.1007/s12668-014-0131-z\u003c/li\u003e\n\u003cli\u003eGhosh V, Ranjha R, Gupta AK. Polymeric encapsulation of anti-larval essential oil nanoemulsion for controlled release of bioactive compounds. \u003cem\u003eInorg Chem Commun\u003c/em\u003e. \u003cstrong\u003e150\u003c/strong\u003e:110507 (2023). Doi:10.1016/j.inoche.2023.110507\u003c/li\u003e\n\u003cli\u003eHeydari M, Amirjani A, Bagheri M, Sharifian I, Sabahi Q. Eco-friendly pesticide based on peppermint oil nanoemulsion: Preparation, physicochemical properties, and its aphicidal activity against cotton aphid. \u003cem\u003eEnviron Sci Poll Res \u003c/em\u003e\u003cstrong\u003e27\u003c/strong\u003e(6):6667-6679 (2020). Doi: 10.1007/s11356-019-07332-y\u003c/li\u003e\n\u003cli\u003eKhan MH, Dar NA, Alie BA, Dar SA, Lone AA, Mir GH \u003cem\u003eet al.\u003c/em\u003e Unraveling the variability of essential oil composition in different accessions of \u003cem\u003eBunium persicum\u003c/em\u003e collected from temperate micro-climates. \u003cem\u003eMolecules\u003c/em\u003e.\u003cstrong\u003e28\u003c/strong\u003e(5):2404 (2023). Doi:10.3390/molecules28052404\u003c/li\u003e\n\u003cli\u003eKoroleva MY, Yurtov EV. Ostwald ripening in macro- and nanoemulsions. \u003cem\u003eRuss Chem Rev\u003c/em\u003e. \u003cstrong\u003e\u003cem\u003e90\u003c/em\u003e\u003c/strong\u003e(3):293 (2021). Doi:10.1070/RCR4962\u003c/li\u003e\n\u003cli\u003eKoundal R, Dolma SK, Chand G, Agnihotri VK, Reddy SE\u003cem\u003e.\u003c/em\u003e Chemical composition and insecticidal properties of essential oils against diamondback moth (\u003cem\u003ePlutella xylostella\u003c/em\u003e L.). \u003cem\u003eToxin review\u003c/em\u003e\u003cstrong\u003e. 41\u003c/strong\u003e(1):48-54 (2020). Doi:10.1080/15569543.2018.1536668 \u003c/li\u003e\n\u003cli\u003eLenth R. Emmeans: estimated marginal means, aka least-squares means. 2023 [Internet]. Available from: https://CRAN.R-project.org/package=emmeans \u003c/li\u003e\n\u003cli\u003eLima LA, Ferreira-S\u0026aacute; PS, Garcia Jr MD, Pereira VLP, Carvalho JCT, Rocha L. Nanoemulsions of the essential oil of \u003cem\u003eBaccharis reticularia\u003c/em\u003e and its constituents as eco-friendly repellents against \u003cem\u003eTribolium castaneum\u003c/em\u003e. \u003cem\u003eInd Crops Prod.\u003c/em\u003e\u003cstrong\u003e162\u003c/strong\u003e:113282 (2021). Doi:10.1016/j.indcrop.2021.113282\u003c/li\u003e\n\u003cli\u003eLiu Z, Li QX, Song B. Pesticidal activity and mode of action of monoterpenes.\u003cem\u003eJ Agric Food Chem\u003c/em\u003e.\u003cstrong\u003e70\u003c/strong\u003e(15):4556-71 (2022). Doi:10.1021/acs.jafc.2c00635\u003c/li\u003e\n\u003cli\u003eLopez L, Liburd OE. Injury to southern highbush blueberries by southern red mites and management using various miticides. \u003cem\u003eInsects.\u003c/em\u003e\u003cstrong\u003e11\u003c/strong\u003e(4):233 (2020). Doi:10.3390/insects11040233 \u003c/li\u003e\n\u003cli\u003eMarucci RC, Ruber SE, Pec M, Liburd, OE. Are predatory mites effective as biological control agents to suppress \u003cem\u003eOligonychus ilicis\u003c/em\u003e (Acari: Tetranychidae) in blueberry plantings?. \u003cem\u003eJ Economic Entom\u003c/em\u003e.\u003cstrong\u003e117\u003c/strong\u003e(3):834-842 (2024). Doi:10.1093/jee/toae086\u003c/li\u003e\n\u003cli\u003eMcClements DJ. Nanoemulsions versus microemulsions: terminology, differences, and similarities. \u003cem\u003eSoft Matter\u003c/em\u003e.\u003cstrong\u003e8\u003c/strong\u003e(6):1719-29 (2012). Doi:10.1039/C2SM06903B\u003c/li\u003e\n\u003cli\u003eModafferi A, Urbaneja A, Aure CM, Laudani F, Palmeri V, Giunti G, P\u0026eacute;rez-Hedo M. Green pest control strategies: essential oil-based nano-emulsions for \u003cem\u003eDelottococcus aberiae\u003c/em\u003e management. \u003cem\u003eJ Pest Sci\u003c/em\u003e\u003cstrong\u003e98\u003c/strong\u003e(3):1263-1275 (2025) Doi:10.1007/s10340-025-01914-1\u003c/li\u003e\n\u003cli\u003eNetter L, Silva YLL, Silva AB, Santos LV, Rocha, SBN. Atividade inseticida do \u0026oacute;leo essencial de hortel\u0026atilde;-pimenta sobre lagartas de \u003cem\u003eSpodoptera frugiperda\u003c/em\u003e. Rev \u003cem\u003eCi\u0026ecirc;nc Agr\u0026iacute;c\u003c/em\u003e.\u003cstrong\u003e22\u003c/strong\u003e(Esp):1-5 (2024). Doi:10.28998/rca.22\u003c/li\u003e\n\u003cli\u003eOca-\u0026Aacute;valos JMM, Candal RJ, Herrera ML. Nanoemulsions: stability and physical properties\u003cem\u003e. Curr Opin Food Sci. \u003c/em\u003e\u003cstrong\u003e16\u003c/strong\u003e:1-6 (2017). Doi:10.1016/j.cofs.2017.06.003 \u003c/li\u003e\n\u003cli\u003eOrtiz-Zamora L, Bezerra DC, de Oliveira HNS, Duarte JL, Guisado-Bourzac F, Chil-N\u0026uacute;\u0026ntilde;ez I. Preparation of non-toxic nano-emulsions based on a Brazilian plant species through a low-energy concept. \u003cem\u003eInd Crops Prod\u003c/em\u003e. \u003cstrong\u003e158\u003c/strong\u003e:112989 (2020). Doi:10.1016/j.indcrop.2020.112989. \u003c/li\u003e\n\u003cli\u003eOzman SS, Sullivan G, Cakir S, Bas H, Saglam D, Doker I, \u003cem\u003eet al.\u003c/em\u003e Phytoseiid mites: Trees, ecology and conservation. \u003cem\u003eDiversity\u003c/em\u003e. \u003cstrong\u003e16\u003c/strong\u003e(9): 542 (2024). Doi:10.3390/d16090542 \u003c/li\u003e\n\u003cli\u003ePaz Neto ADA, da C\u0026acirc;mara CAG, Monteiro VB, de Moraes MM, de Melo JPR, Leal TTB\u003cem\u003e.\u003c/em\u003e Bioactivity of essential oils from three species of \u003cem\u003eMentha \u003c/em\u003eL. against \u003cem\u003ePlutella xylostella\u003c/em\u003e (Linnaeus, 1767)(Lepidoptera: Plutellidae). \u003cem\u003eEntomologia Neotrop.\u003c/em\u003e\u003cstrong\u003e54\u003c/strong\u003e(1):101 (2025). Doi:10.1007/s13744-025-01317-y \u003c/li\u003e\n\u003cli\u003ePiffer ABM, Holtz AM, Neto VB, de Paula Marchiori JJ, de Carvalho JR, Pretti IR, \u003cem\u003eet al.\u003c/em\u003e Can soil conditioners of plant origin be used for pest control? \u003cem\u003eObserv Econ Lat Am \u003c/em\u003e\u003cstrong\u003e21\u003c/strong\u003e(10): 15883\u0026ndash;15900 (2023). Doi:10.55905/oelv21n10-075 \u003c/li\u003e\n\u003cli\u003eR Core Team. R: a language and environment for statistical computing. 2023 [Internet]. Available from: http://www.R-project.org\u003c/li\u003e\n\u003cli\u003eReis PR, Alves EB, Sousa EO. Biologia do \u0026aacute;caro-vermelho do cafeeiro \u003cem\u003eOligonychus ilicis\u003c/em\u003e (McGregor, 1917). \u003cem\u003eCi\u0026ecirc;ncia e Agrotecnologia\u003c/em\u003e, \u003cstrong\u003e21\u003c/strong\u003e: 260-266 (1997). Doi:10.5555/19981110067\u003c/li\u003e\n\u003cli\u003eRichardson LL, Adler LS, Leonard AS, Andicoechea J, Regan KH, Anthony WE, \u003cem\u003eet al.\u003c/em\u003e Secondary metabolites in floral nectar reduce parasitic infections in bumblebees. \u003cem\u003eProc Biol Sci\u003c/em\u003e. \u003cstrong\u003e282\u003c/strong\u003e(1803):20142471 (2015). Doi:10.1098/rspb.2014.2471\u003c/li\u003e\n\u003cli\u003eSantos CAL, Moreira AMT, Teles BRDS, Kamdem JP, AlAsmari AF, Alasmari F. \u003cem\u003eet al\u003c/em\u003e. \u003cem\u003eMentha arvensis\u003c/em\u003e oil exhibits repellent, acute toxic, and antioxidant activities in \u003cem\u003eNauphoeta cinerea\u003c/em\u003e. \u003cem\u003eSci Rep\u003c/em\u003e.\u003cstrong\u003e14\u003c/strong\u003e(1):21599 (2024). Doi:10.1038/s41598-024-72722-3\u003c/li\u003e\n\u003cli\u003eSouza LP, Zuim V, Stinguel P, Pinheiro PF, Zago HB. Toxicity of essential oil of \u003cem\u003eMentha piperita\u003c/em\u003e (Lamiaceae) and its monoterpenoid menthol against \u003cem\u003eTetranychus urticae\u003c/em\u003e Kogan 1836 (Acari: Tetranychidae). \u003cem\u003eAn Acad Bras Cienc.\u003c/em\u003e\u003cstrong\u003e94\u003c/strong\u003e(4):e20200427 (2022). Doi:10.1590/00013765202220200427 \u003c/li\u003e\n\u003cli\u003eSouza RB, Guimar\u0026atilde;es JR. Effects of avermectins on the environment based on its toxicity to plants and soil invertebrates\u0026mdash;a review.\u003cem\u003eWater Air Soil Pollut\u003c/em\u003e. \u003cstrong\u003e233\u003c/strong\u003e(7):259 (2022). Doi:10.1007/s11270-022-05744-0\u003c/li\u003e\n\u003cli\u003eStringaro A, Colone M, Angiolella L. Antioxidant, antifungal, antibiofilm and cytotoxic activities of Mentha spp essential oils. \u003cem\u003eMedicines\u003c/em\u003e. \u003cstrong\u003e5\u003c/strong\u003e(4):112 (2018). Doi:0.3390/medicamentos5040112 \u003c/li\u003e\n\u003cli\u003eWu Z, Jin C, Chen Y, Yang S, Yang X, Zhang D, Xie Y. Essential oils of \u003cem\u003eMentha spp\u003c/em\u003e. as a potential toxic fumigant with acetylcholinesterase inhibition in \u003cem\u003eReticulitermes dabieshanensis\u003c/em\u003e. \u003cem\u003ePlants.\u003c/em\u003e\u003cstrong\u003e12\u003c/strong\u003e(23):4034 (2023). Doi:10.3390/plants12234034\u003c/li\u003e\n\u003cli\u003eZhang W, Jiang H, Rhim JW, Cao J, Jiang W. Effective strategies of sustained release and retention enhancement of essential oils in active food packaging films/coatings\u003cem\u003e. Food Chem.\u003c/em\u003e\u003cstrong\u003e367:\u003c/strong\u003e130671 (2022). Doi:10.1016/j.foodchem.2021.130671\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Table","content":"\u003cp\u003eTable 2 is available in the Supplementary Files section.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"bionanoscience","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bnsc","sideBox":"Learn more about [BioNanoScience](http://link.springer.com/journal/12668)","snPcode":"12668","submissionUrl":"https://submission.nature.com/new-submission/12668/3","title":"BioNanoScience","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"alternative pest control, biopesticide, Coffea canephora, phytochemicals, red coffee mite","lastPublishedDoi":"10.21203/rs.3.rs-9260778/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9260778/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe red mite, \u003cem\u003eOligonychus ilicis\u003c/em\u003e McGregor, 1917 (Acari: Tetranychidae), is a major pest of coffee crops (\u003cem\u003eCoffea canephora\u003c/em\u003e), causing significant damage to plantations. In this context, essential oils, particularly when formulated as nanoemulsions, emerge as a promising alternative for pest management due to their high efficacy and low environmental toxicity. This study explores the acaricidal potential of a nanoemulsion based on \u003cem\u003eMentha piperita\u003c/em\u003e essential oil against \u003cem\u003eO. ilicis\u003c/em\u003e. For this study, an analysis of the main chemical constituents of the essential oil (EO) was first carried out, and a nanoemulsion based on peppermint essential oil was formulated using the low-energy emulsification method, with specific surfactants employed to stabilize the particles. The proposed formulation remained stable over time, and subsequently, the toxicity of the nanoemulsion was evaluated through laboratory bioassays involving the direct application of different concentrations (0.025, 0.040, 0.060, 0.10, 0.15, and 0.25% [mL mL⁻\u0026sup1;]) to adult females of \u003cem\u003eO. ilicis\u003c/em\u003e. The results revealed a dose-dependent mortality, with mortality rates exceeding 90% for \u003cem\u003eO. ilicis\u003c/em\u003e at concentrations as low as 0.040%. The LC₅₀ and LC₉₀ values were estimated at 0.0257 and 0.0424% (mL mL⁻\u0026sup1;), respectively. These findings demonstrate that the peppermint essential oil-based nanoemulsion exhibits strong acaricidal activity against the red coffee mite, \u003cem\u003eO. ilicis\u003c/em\u003e. Given its efficacy and potential environmental safety, this formulation represents a promising and sustainable tool for the integrated management of this key pest in coffee production systems.\u003c/p\u003e","manuscriptTitle":"Nanoemulsion based on essential oil of Mentha piperita in the sustainable management of Oligonychus ilicis McGregor, 1917 (Acari: Tetranychidae)","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-04-16 15:00:06","doi":"10.21203/rs.3.rs-9260778/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-04-20T20:20:32+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-20T09:22:15+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-17T01:45:13+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"334131150302540894662703417373125320214","date":"2026-04-15T07:55:49+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-11T22:31:07+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"213541598635051100964057617190012467299","date":"2026-04-11T21:48:22+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"18263158331895151685487509309282485090","date":"2026-04-10T17:10:04+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"264202433334016326185839994236674539160","date":"2026-04-09T10:48:23+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"298721942695232652867440228144024624095","date":"2026-04-09T01:40:05+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-04-08T13:08:34+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-04-08T11:17:35+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-04-08T09:30:31+00:00","index":"","fulltext":""},{"type":"submitted","content":"BioNanoScience","date":"2026-03-29T19:45:27+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bionanoscience","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bnsc","sideBox":"Learn more about [BioNanoScience](http://link.springer.com/journal/12668)","snPcode":"12668","submissionUrl":"https://submission.nature.com/new-submission/12668/3","title":"BioNanoScience","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"a7954648-743c-4bcb-a715-9c97ec214898","owner":[],"postedDate":"April 16th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-05-07T15:53:56+00:00","versionOfRecord":[],"versionCreatedAt":"2026-04-16 15:00:06","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-9260778","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-9260778","identity":"rs-9260778","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}