Nanoemulsion Based on Piper tuberculatum Essential Oil for the Treatment of the Crop Pest Bemisia tabaci Biotype B

Abstract

The agricultural sector has been searching for eco-friendly methods for pest control. Essential oil-based formulations are promising alternatives. Here, we developed a nanoemulsion based on Piper tuberculatum Jacq. essential oil to control Bemisia tabaci biotype B. The essential oils extracted from P. tuberculatum leaves were mostly composed of β-caryophyllene (16.86%), germacrene D (13.39%), and α-pinene (11.60%). Nanoemulsions containing essential oils were significantly toxic to B. tabaci adults 24 h after cabbage leaf treatment. The lethal concentration required to reduce the number of individuals by 50% (LC₅₀) was 8700 ppm, with a confidence interval (95% CI) of 7900-8800 ppm. Notably, nanoemulsions with 10000 and 20000 ppm essential oils presented insecticidal activity statistically similar to that of commercial synthetic insecticides. Our findings highlight the innovative potential of P. tuberculatum essential oil nanoemulsions as effective alternatives for controlling B. tabaci, providing a sustainable and economically viable solution.

Keywords:
whitefly; nanotechnology; nanoformulation; natural products

Introduction

Bemisia tabaci Gennadius biotype b (Hemiptera: Aleyrodidae), commonly known as whitefly, is a global pest that affects several agricultural crops. By feeding on the vascular tissues of plants, sap-sucking insects play an additional role by serving as transmission vectors for several phytopathogenic viruses.¹ Chemical control has been widely used in the management of B. tabaci; however, its indiscriminate use is associated with environmental damage and human health, in addition to causing the development of resistance and affecting nontarget species.² In this sense, alternative and sustainable methods, such as the use of botanical insecticides, have gained prominence.³

Botanical insecticides contain compounds resulting from the secondary metabolism of plants, which are part of their chemical defense against herbivorous insects.⁴ Insecticidal active ingredients can come from the whole plant or parts of it and can be powders, extracts, or essential oils.⁵ This alternative method has gained considerable attention because of its low residual effect and low toxicity for mammals, natural enemies, and pollinators, as well as greater selectivity,⁶ constituting an integrated pest management (IPM) methodology.⁷

Essential oils are isolated from plants as mixtures of volatile, lipophilic, and strongly odorous secondary metabolites, mainly terpenoids such as monoterpenes and sesquiterpenes.⁸ They exhibit repellent properties, insecticide, growth inhibitor, oviposition inhibitor, ovicidal, and growth-reducing effects on a variety of insects. The great structural diversity of its components makes it a potential source of multitarget bioactive molecules. Its components can disrupt the waxes and cuticular membranes of insect integuments, leading to desiccation, and act on digestive and neurological enzymes of the insects,⁹ favoring their use in the control of agricultural pests such as B. tabaci.

Despite their potential, the use of essential oils faces limitations due to their high volatility, low solubility in water, susceptibility to oxidation and instability in the face of reactions caused by light, humidity, and temperature. An alternative to improve the stability and effectiveness of these oils is the use of nanotechnology aimed at encapsulating their bioactive compounds.¹⁰ Among the various nanostructured systems are nanoemulsions, which are colloidal systems whose particles have dimensions on the order of 20-500 nm and function as vehicles for active molecules.¹¹, ¹² This provides characteristics such as a transparent visual appearance, high colloidal stability, and an extensive interfacial area with respect to volume. Nanoformulations represent an emerging technology with broad applications. Features such as increased efficiency, durability, and a reduced effective concentration can result in significant benefits, increasing the value of nanostructured products.¹³

Plants from the Piperaceae family constitute a source of essential oils with insecticidal activity, especially species from the Piper genus.¹⁴ Among these species, Piper tuberculatum Jacq. has demonstrated a potential role in controlling insect pests.¹⁴, ¹⁵ This species is widely distributed in tropical regions around the world and is found in several Latin American countries, including Brazil, Colombia and Venezuela.¹⁶ An interesting characteristic of these plants is the accumulation of several physiologically active natural products, such as alkaloids and amides.¹⁷ However, to the best of our knowledge, there are no reports on the development of a formulation based on P. tuberculatum essential oil nanoemulsions.

The present study aimed to develop a nanoemulsion-based formulation containing P. tuberculatum essential oil to control B. tabaci. Initially, the essential oil was extracted from the leaves of P. tuberculatum, and its chemical composition was characterized via gas chromatography coupled with mass spectrometry (GC-MS). The nanoemulsions were subsequently formulated with different concentrations of the essential oil and tested via in vitro bioassays with adult B. tabaci.

Experimental

Botanical material

Leaves from the aerial part of P. tuberculatum were collected in the morning on the banks of the Gurguéia River, Bom Jesus, PI, Brazil (9°04’46.2”S 44°19’33.9”W −9.079491, −44.326091). The exsiccate was deposited in the Graziela Barroso Herbarium under number TEPB 32523. The plants were registered in the National System for Management of Genetic Heritage and Associated Traditional Knowledge (SisGen) under number AE123A5.

Extraction and chemical analysis of essential oils

The leaves were dried in an air circulation oven at 45 °C for 72 h. After drying, the material was crushed in a knife mill (Solab, SL-31, SP, Brazil), and 150 g of leaf powder was mixed with 1 L of distilled water and subjected to a hydrodistillation process for 4 h via a Clevenger-type extractor. The crude essential oil formed during hydrodistillation was collected with a Pasteur pipette, and the yield was calculated.

The essential oils were analyzed in triplicate on a gas chromatograph coupled with a mass spectrometer (Shimadzu GCMS-QP2010 SE, AOC-5000, Japan) operated by electron impact at 70 eV. The chromatograph was equipped with a fused silica capillary column (DB5, 30 m × 0.25 mm × 0.25 µm, Agilent) using high-purity helium as the carrier gas with a flow rate of 1 mL min¹. The injector and interface temperatures were 260 °C. The temperature program ranged from 50 to 260 °C at 12 °C min⁻¹ and was held for 5 min. The samples were diluted in dichloromethane (Neon, SP, Brazil) (1 μg mL⁻¹), and 1 µL was injected in 10:1 split mode. Mass spectra were recorded in the range of m/z 50-400 at 3.54 scans s⁻¹. The mass spectra of the compounds were compared with those of the Nist library database. The samples were injected with a standard mixture of n-alkanes (C7-C40, Sigma-Aldrich, SP, Brazil) and the retention index (RI) calculated according to the van den Dool and Kratz equation.¹⁸

Preparation and characterization of nanoemulsions

The nanoemulsions were prepared via emulsion phase inversion (EPI) following the low-energy protocol of Ostertag et al.¹⁹ modified. The experiments were designed to obtain a final composition with a total mass of 50 g, including deionized water (m/m), tween 80 (Dinâmica, SP, Brazil) (5000 ppm, m/m), and essential oils at different concentrations (0; 5000; 10000; 20000 ppm, m/m). The organic phase was prepared by mixing the essential oil and surfactant under constant stirring at 750 rpm for 30 min. Subsequently, the aqueous phase was added to the organic phase via a burette with a flow rate of 2 mL min⁻¹, while the system was maintained under constant stirring (750 rpm) for 60 min. The formulations were prepared in triplicate for each dose, maintained at room temperature (30 ± 2 °C), and characterized after 1 and 30 days via dynamic light scattering (DLS) (nanoPartica SZ-100V2, Horiba Scientific, USA) to determine the size of the droplets.

B. tabaci rearing and toxicity bioassay

Biological toxicity tests were performed on B. tabaci adults. The insects were collected in Eugenópolis, PI, Brazil (9°12’58”S 44°26’37”W) and maintained in a greenhouse with cabbage plants (Brassica oleracea L.). To set up the bioassay, 90-day-old cabbage leaves were removed with a scalpel, leaving a 5 cm petiole. The leaves were subsequently immersed in the following treatments: (i) nanoemulsion of P. tuberculatum essential oil at different concentrations (0, 5000, 10000, or 20000 ppm) or (ii) the synthetic insecticide thiamethoxam prepared according to the product label (250000 ppm m/m). The treated leaves were placed in Petri dishes containing filter paper moistened with distilled water. Additionally, the petioles were covered with cotton, which was moistened daily with distilled water. B. tabaci adults (10 individuals) were transferred to Petri dishes containing treated cabbage leaves and then incubated in a biochemical oxygen demand (B.O.D.) incubator (Lucadema, Brazil) at 27 °C, with a 12 h photoperiod.⁶

The percentage of insect mortality was recorded 24 h posttreatment and calculated according to the following equation: percentage mortality = (number of individuals killed/number of individuals treated) × 100.²⁰ The design was completely randomized, with four replications for each dose. The data were subjected to the Mann-Whitney nonparametric test followed by regression analysis via SigmaPlot v14.0 software.²¹ The lethal concentration required to reduce the number of individuals by 50% (LC₅₀) and 95% confidence intervals (95% CI) were estimated from the regression equation.

Results and Discussion

Chemical composition of P. tuberculatum essential oil

The yield of essential oil from P. tuberculatum leaves was 0.16 ± 0.05%. A total of 36 compounds were identified through GC-MS analyses (Table 1), with β-caryophyllene (16.86%), germacrene D (13.39%), and α-pinene (11.60%) as the major constituents. These compounds were also found in the essential oil from P. tuberculatum leaves collected in São Paulo, Brazil, at concentrations of 40.2, 10.4, and 5.5%, respectively.²² β-Caryophyllene was also the main compound in the essential oil of P. tuberculatum leaves collected in Amazonas, Brazil (54.8%), while 8.42% were oxygenated sesquiterpenes.²³ The major compounds in the essential oils of P. tuberculatum leaves collected in Ceará, Brazil, were α-pinene (11.27%), β-pinene (20.01%), 1,8-cineole (10.81%), linalool (28.61%), and other compounds, including germacrene D (3.43%).²⁴ Sesquiterpenoids and oxygenated derivatives were found in the essential oil from P. tuberculatum leaves collected in Sucre, Venezuela, with spathulenol (11.37%) as the main constituent.²⁵ Leaves of P. tuberculatum collected in Rondônia, Brazil, are rich in alloaromadendrene, γ-cadinene, germacrene D-4-ol, nerolidol, and carvone.²⁶

Table 1
Chemical composition of the essential oil from P. tuberculatum leaves

No.	Compound	RI (calc.)	RI (lit.)	Relative abundancea / %
1	α-pinene	925	933	11.60 ± 2.10
2	camphene	972	953	0.06 ± 0.01
3	sabinene	983	976	0.82 ± 0.12
4	β-pinene	977	980	6.33 ± 0.79
5	β-myrcene	989	991	0.47 ± 0.08
6	pseudolimonene	–	–	0.03 ± 0.01
7	origanene	–	–	0.04 ± 0.01
8	o-cymene	1025	1020	0.03 ± 0.02
9	limonene	1029	1036	8.65 ± 2.10
10	β-cymene	–	–	1.71 ± 0.07
11	3-carene	–	–	0.02 ± 0.01
12	terpinolene	–	–	0.05 ± 0.01
13	γ-elemene	–	–	0.09 ± 0.02
14	δ-elemene	1336	1337	0.32 ± 0.14
15	α-cubebene	1348	1345	0.06 ± 0.01
16	copaene	1377	1376	3.54 ± 0.29
17	β-caryophyllene	1408	1428	16.86 ± 0.89
18	α-caryophyllene	1457	1454	1.26 ± 0.13
19	aromadendrene	1462	1464	1.79 ± 0.18
20	cycloisosativene	1480	1479	0.04 ± 0.01
21	germacrene D	1483	1480	13.39 ± 1.74
22	eremofilene	1491	–	0.18 ± 0.07
23	germacrene b	–	–	8.49 ± 0.95
24	β-bisabolene	1508	1509	0.37 ± 0.06
25	δ-cadinene	1519	1524	1.85 ± 0.64
26	nerolidol	1561	1564	4.12 ± 0.54
27	humulen-(v1)	1597	1505	0.55 ± 0.07
28	spathulenol	1579	1575	2.54 ± 0.10
29	caryophyllene oxide	1585	1581	2.09 ± 0.21
30	cubenol	1623	1616	0.48 ± 0.03
31	tau-muurolol	1646	1642	0.99 ± 0.06
32	α-cadinol	1658	1653	1.06 ± 0.04
33	β-eudesmol	–	–	1.44 ± 0.49
34	homomenthyl salicylate	–	–	0.11 ± 0.04
35	nerolidyl acetate	–	–	0.07 ± 0.02
36	unknown compounds			8.51

• ^aAverage (relative abundance in percentage) ± standard deviation. RI (calc.): calculated retention index; RI (lit.): retention index from the literature.²⁷

Leaves of P. tuberculatum var. tuberculatum (Micq.) collected in Rondônia, Brazil, revealed that β-caryophyllene (26.3%) and α-cadinol (13.7%) were the main components of the essential oil.²⁸ Germagrene D (36.32%) was identified as the major compound in the essential oil from P. tuberculatum var. minus leaves collected in Rio de Janeiro, Brazil.²⁹ Therefore, there is a predominance of monoterpenes and sesquiterpenes in the essential oil of the leaves of this species.²³, ²⁵, ³⁰ Additionally, the major constituents of the essential oils of the fruits and fine stems of P. tuberculatum collected in Rondônia, Brazil, were caryophyllene oxide (32.1-26.6%) and (E)-caryophyllene (17.7-12.3%),³¹ whereas fruits collected in Ceará, Brazil, were composed mainly of β-pinene (27.74-31.51%), α-pinene (26.54-28.38%), and Z-ocimene (20.22%).³²

These findings suggest that although essential oils from aromatic plants have similar chemical compositions, the specific proportions differ. This variation can be attributed to species chemotaxonomy, leaf age, geographic region, seasonality, and different responses to the influence of environmental factors, such as temperature, pressure, the presence of predators and nutrient availability.³³ The similarity in chemical composition, even with different proportions, may indicate the need for these plants to produce similar secondary metabolites for specific ecological interactions. Therefore, it is possible to infer that plants respond adaptively to the environment, developing chemical compositions that meet the specific demands of their ecology.³⁴

Nanoemulsion of essential oil from P. tuberculatum leaves

Nanoemulsions prepared from P. tuberculatum essential oil had average particle sizes between 11 and 185 nm from 1 to 30 days after preparation (Table 2). Therefore, P. tuberculatum essential oil is favorable for the formation of stable nanoemulsions with droplet sizes smaller than 200 nm.³⁵ Additionally, the particle size of the nanoemulsion increased as the concentration of essential oil increased, which was attributed to the greater amount of active ingredient encapsulated in the colloidal system.

Table 2
DLS of the nanoemulsion loaded with P. tuberculatum essential oil at different doses after 1 and 30 days of preparation

Concentration / ppm	Droplet size (1 day) / nm	Droplet size (30 days) / nm
0	8.85 ± 1.69	8.43 ± 1.63
5000	11.00 ± 2.44	13.46 ± 3.26
10000	164.69 ± 11.36	185.05 ± 26.54
20000	163.73 ± 30.92	179.93 ± 39.40

Nanoemulsions significantly improve the solubility of lipophilic substances present in essential oils, which provides benefits for agriculture. Among the advantages, the best diffusion of active components on the treated surfaces stands out, improving the power of penetration into tissues due to low surface tension and greater surface area, consequently providing an acute effect.³⁶, ³⁷ The optical properties of the nanoemulsion, such as its transparency, are also fundamental for several applications. The smaller the particle size is, the greater the transparency of the sample, providing greater stability against aggregation and better bioavailability of the components.³⁸ Furthermore, nanoemulsions guarantee the controlled and gradual release of the active ingredient at the target location, which reduces the need to use high concentrations of essential oil.³⁹

Thus, the active ingredient is released in a controlled manner, under specific conditions or through external stimuli, whether by chemical cleavage or physical diffusion. However, formulations based on natural products, such because essential oils, have the advantage of preventing loss of effectiveness due to processes such as evaporation, because essential oils are quite volatile. This results in a significant improvement in the interaction with the insect pest and, consequently, an increase in pesticide activity.³⁷, ³⁸ The formulation of nanoemulsions also promotes the photoprotection of essential oils for application under field conditions since their instability in the presence of light, heat, and humidity leads to numerous degradation reactions, altering their chemical composition and bioactivity.⁴⁰

Nanoemulsions can be prepared via different techniques; however, low-energy methods, which harness the internal chemical energy of the system, are often more efficient and generally allow the production of smaller droplet sizes than high-energy methods.⁴¹ The low-energy technique made it possible to use simple equipment, which not only facilitated the process but also helped to preserve the physical-chemical properties of the essential oil, maintaining its effectiveness and original characteristics. Furthermore, the absence of organic solvents in this approach makes it environmentally friendly, making this method suitable for large-scale production. The surfactant/emulsifier tween 80 used in the formulation allowed the formation and effective stabilization of the colloidal system,¹⁹, ⁴¹ promoting a high value of hydrophilic-lipophilic balance, which reduced the tension between the aqueous and oily phases, resulting in a stable nanoemulsion. Nonequilibrium nanoemulsions generally seek to reduce their interfacial free energy, which can lead to problems such as creaming, flocculation, sedimentation, Ostwald ripening, and coalescence.¹⁹

Toxicity of P. tuberculatum in natura essential oil and nanoemulsions to B. tabaci adults

P. tuberculatum essential oil nanoemulsions were toxic to B. tabaci adults 24 h after treating the cabbage leaves, with a significant difference between the concentrations evaluated (p = 0.006). The dose-response curves were fitted to a logistic function (Figure 1), with nanoemulsions at doses of 10000 and 20000 ppm P. tuberculatum essential oil showing statistical similarity to the synthetic insecticide thiamethoxam (Figure 2), whereas the dose of 5000 ppm did not differ from the control treatment. The lethal concentration of nanoemulsions required to reduce the number of individuals by 50% (LC₅₀) was 8700 ppm, with a confidence interval (95% CI) of 7900-8800 ppm. P. tuberculatum in natura essential oil was toxic to B. tabaci adults only at 20000 ppm (Figure 2); however, it caused phytotoxicity to cabbage leaves (data not shown).

Figure 1
Mortality of B. tabaci adults 24 h after treatment of cabbage leaves with P. tuberculatum in natura essential oil and nanoemulsions at different doses.

Figure 2
Mortality of B. tabaci adults in response to each dose of P. tuberculatum in natura essential oil and nanoemulsions in relation to the control treatments and the synthetic insecticide. The bars in the figures represent the standard deviation of the repetitions. Treatments with the same letter are significantly different according to the Mann-Whitney test at the 0.05 level.

P. tuberculatum essential oil nanoemulsions tested against B. tabaci adducts at different concentrations showed excellent results in terms of their effectiveness in controlling the pest, but at the same time, they did not have phytotoxic effects on plants. Our study represents a pioneering contribution involving the use of nanoemulsions based on P. tuberculatum essential oil to control insect pests, although other Piper species have been used for the synthesis of nanoemulsions.⁴², ⁴³, ⁴⁴ This gap in research is particularly relevant given the increasing resistance of B. tabaci to conventional commercial insecticides, probably due to the use of a single active ingredient with insecticidal activity.²

Previous reports have described the toxicity of P. tuberculatum essential oil to adults of Cerotoma arcuatus (Oliv.) (Crysomelidae),⁴⁵ second-instar nymphs of the red cowpea bug Crinocerus sanctus (Fabr.) (Hemiptera; Coreidea),⁴⁶ and rice stalk stink bug eggs, Tibraca limbativentris (Hemiptera: Pentatomidae).⁴⁷ The essential oil from P. tuberculatum leaves and β-caryophyllene exhibited insecticidal activity against Aedes aegypti (Culicidae) larvae and showed no lethality toward nontarget organisms, especially Toxorhynchites haemorrhoidalis (Culicidae), Anisops bouvieri (Hemiptera), and Diplonychus indicus (Heteroptera).²³ However, despite its insecticidal potential, there are no records of the use of P. tuberculatum essential oil for pest control in the context of nanoformulations. This highlights the need to explore this approach, such as nanoemulsions, to improve the efficacy and applicability of these compounds.

The toxicity of essential oils to insects is influenced by their chemical composition, which depends on external factors.⁴⁸ In contrast to commercial synthetic pesticides, bioactive natural compounds can act via multiple mechanisms, making the development of resistance less likely due to the diversity of active ingredients present in each mixture.⁴⁹ Furthermore, the size of the nanometer-scale particles increases the effectiveness of the essential oil by up to 20 times to reach different parts of the insect’s body after ingestion, resulting in death in less time.⁴² The essential oil can also act by inhibiting the enzyme acetylcholinesterase, which leads to the collapse of the nervous system due to the uncontrolled transmission of nerve impulses, causing accelerated movements, tremors, lethargic movements and paralysis followed by death.⁹

Conclusions

The essential oil from P. tuberculatum leaves was characterized mainly by the presence of monoterpenes and sesquiterpenes. Nanoemulsions prepared from the essential oil of P. tuberculatum leaves have proven to be promising eco-friendly pesticides for the control of B. tabaci adults. These findings demonstrate the feasibility of continuing studies with this bioformulation under field conditions in association with integrated pest management practices.

Acknowledgments

The authors acknowledge the Fundação Universidade Federal do Piauí (FUFPI), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Finance Code 001), and Centro de Educação Aberta e a Distância (CEAD) for financial support.

References

• ¹ Abubakar, M.; Koul, B.; Chandrashekar, K.; Raut, A.; Yadav, D.; Agriculture 2022, 12, 1317 [Crossref]; Perring, T. M.; Crop Prot. 2001, 9, 725. [Crossref]
  » Crossref» Crossref
• ² Palumbo, J. C.; Horowitz, A. R.; Prabhaker, N.; Crop Prot. 2001, 20, 739 [Crossref]; Horowitz, A. R.; Ghanim, M.; Roditakis, E.; Nauen, R.; Ishaaya, I.; J. Pest. Sci. 2020, 93, 893. [Crossref]
  » Crossref» Crossref
• ³ Rattan, R. S.; Prot. Crop 2010, 29, 913. [Crossref]
  » Crossref
• ⁴ Miresmaill, S.; Isman, M. B.; Trends Plant Sci. 2014, 19, 29. [Crossref]
  » Crossref
• ⁵ Campos, E. V. R.; Proença, P. L. F.; Oliveira, J. L.; Bakshi, P. C. B.; Fraceto, L. F.; Ecol. Indic. 2019, 105, 483. [Crossref]; Hikal, W. M.; Baeshen, R. S.; Ahl, H. A. H. S. A.; Cogent Biol. 2017, 3, 1. [Crossref]
  » Crossref» Crossref
• ⁶ Iqbal, N.; Hazra, D. K.; Purkait, A.; Agrawal, A.; Kumar, J.; Colloids Surf. B 2022, 209, 112176. [Crossref]; Chaudhari, A. K.; Singh, V. K.; Kedia, A.; Das, S.; Kishore, N.; Environ. Sci. Pollut. Res. 2021, 28, 18918. [Crossref]
  » Crossref» Crossref
• ⁷ Zhou, W.; Arcot, Y.; Medina, R. F.; Bernal, J.; Cisneros-Zevallos, L.; Akbulut, M. E. S.; ACS Omega 2024, 9, 41130. [Crossref]
  » Crossref
• ⁸ Regnault-Roger, C.; Vincent, C.; Arnason, J. T.; Annu. Rev. Entomol. 2012, 57, 405. [Crossref]
  » Crossref
• ⁹ Jankowska, M.; Rogalska, J.; Wyszkowska, J.; Stankiewicz, M.; Molecules 2017, 23, 34. [Crossref Crossref]
  » Crossref
• ¹⁰ Sarmah, K.; Anbalagan, T.; Marimuthu, M.; Mariappan, P.; Angappan, S.; Vaithiyanathan, S.; J. Pest Sci. 2024. [Crossref]
  » Crossref
• ¹¹ Gupta, A. In Nanoparticles for Biomedical Applications; Chung, E. J.; Leon, L.; Rinaldi, C., eds.; Elsevier, 2020. [Crossref]
  » Crossref
• ¹² Mustafa, I. F.; Hussein, M. Z.; Nanomaterials 2020, 10, 1608. [Crossref]
  » Crossref
• ¹³ Kumar, S.; Nehra, M.; Dilbaghi, N.; Marrazza, G.; Hassan, A. A.; Kim, K.-H.; J. Controlled Release 2018, 294, 131. [Crossref]
  » Crossref
• ¹⁴ Scott, I. M.; Jensen, H. R.; Philogène, B. J. R.; Arnason, J. T.; Phytochem. Rev. 2008, 7, 65. [Crossref]
  » Crossref
• ¹⁵ Ramos, C. S.; Silva, M. W. F.; Moraes, M. M.;Almeida,V.A.; Vanin, S. A.; Kato, M. J.; J. Braz.. Chem. Soc. 2020, 31, 724. [Crossref]
  » Crossref
• ¹⁶ Mgbeahuruike, E. E.; Yrjönen, T.; Vuorela, H.; Holm, Y.; S. Afr. J. Bot. 2017, 112, 54. [Crossref]
  » Crossref
• ¹⁷ Morandim-Giannetti, A. A.; Pin, A. R.; Pietro, N. A. S.; Oliveira, H. C.; Mendes-Giannini, M. J. S.; Alecio, A. C.; Kato, M. J.; de Oliveira, J. E.; Furlan, M.; J. Med. Plants Res. 2010, 1810. [Crossref]
  » Crossref
• ¹⁸ Van Den Dool, H.; Kratz, P. D. J. A.; J. Chromatogr. A 1963, 11, 463. [file:///C|/DRIVE_D/JOB/SCIELO/000000/Crossref]
  » Crossref
• ¹⁹ Ostertag, F.; Weiss, J.; McClements, D. J.; J. Colloid Interface Sci. 2012, 388, 95. [Crossref]
  » Crossref
• ²⁰ Abbott, W. S.; J. Econ. Entomol. 1925, 18, 265. [Crossref]
  » Crossref
• ²¹ SigmaPlot, v. 14.0; Systat Software Inc., USA, 2018.
• ²² Navickiene, H. M. D.; Morandim, A. A.; Alécio, A. C.; Regasini, L. O.; Bergamo, D. C. B.; Telascrea, M.; Cavalheiro, A. J.; Lopes, M. N.; Bolzani, V. S.; Furlan, M.; Marques, M. O. M.; Young, M. C. M.; Kato, M. J.; Quim. Nova 2006, 29, 467. [Crossref]
  » Crossref
• ²³ Lima, S. C.; de Oliveira, A. C.; Tavares, C. P.; Costa, M. L. L.; Roque, R. A.; Environ. Sci. Pollut. Res. 2024, 31, 33454. [Crossref]
  » Crossref
• ²⁴ Cysne, J. B.; Canuto, K. M.; Pessoa, O. D. L.; Nunes, E. P.; Silveira, E. R.; J. Braz. Chem. Soc. 2005, 16, 1378. [Crossref]
  » Crossref
• ²⁵ Salehi, B.; Gyawali, R.; Salam, A. I.; Shinwari, Z. K.; Khan, T.; Sharifi-Rad, J.; Ozleyen, A.; Turkdonmez, E.; Valussi, M.; Tumer, T. B.; Fidalgo, L. M.; Martorell, M.; Setzer, W. N.; Molecules 2019, 24, 1364. [Crossref]
  » Crossref
• ²⁶ Sobreira, P. H. M.; Hernandez, A. E. F.; Souza, A. C. R.; A., A.; A.; Saber Científico 2021, 6, 20. [Link] accessed in February 2025
  » Link
• ²⁷ Pherobase: Database of Pheromones and Semiochemicals, http://www.pherobase.com, accessed in January 2025
  » http://www.pherobase.com
• ²⁸ Facundo, V. A.; de Morais, S. M.; J. Essent. Oil Res. 2005, 17, 304. [Crossref]
  » Crossref
• ²⁹ dos Santos, P. R. D.; Moreira, D. L.; Guimarães, E. F.; Kaplan, M. A. C.; A.; Phytochemistry 2001, 58, 547. [Crossref]
  » Crossref
• ³⁰ Mesquita, J. M. O.; Cavaleiro, C.; Cunha, A. P.; Lombardi, J. A.; Oliveira, A. B.; Rev. Bras. Farm. 2005, 15, 6. [Crossref]
  » Crossref
• ³¹ Facundo, V. A.; Pollli, A. R.; Rodrigues, R. V.; Militão, J. S. L. T.; Stabelli, R. G.; Cardoso, C. T.; Acta Amazonica 2008, 38, 743. [Crossref]
  » Crossref
• ³² Sales, V. S.; Cabral, F. R.; Sales, E. P. N.; Carvalho, T. B.; Costa, M. H. N.; Oliveira, V. A. P.; Rodrigues, C. K. S.; Figueiredo, F. R. S. D. N.; Bezerra, D. S.; Delmondes, G. A.; Coutinho, H. D. M.; Costa, J. G. M.; Menezes, I. R. A.; Felipe, C. F. B.; Kerntopf, M. R.; FoodBiosci. 2022, 48, 101813. [Crossref]
  » Crossref
• ³³ Figueiredo, A. C.; Barroso, J. G.; Pedro, L. G.; Scheffer, J. J. C.; Flavour Fragrance J. 2008, 23, 213. [Crossref]
  » Crossref
• ³⁴ Regnault-Roger, C.; Integr Pest Manage. Rev. 1997, 2, 25. [Crossref]
  » Crossref
• ³⁵ Sharma, S.; Loach, N.; Gupta, S.; Mohan, L.; Environ. Nanotechnol., Monit. Manage. 2020, 14, 100331. [Crossref]
  » Crossref
• ³⁶ Lucia, A.; Guzmán, E.; Adv. Colloid Interface Sci. 2021, 287, 102330. [Crossref]
  » Crossref
• ³⁷ McClements, D. J.; Rao, J.; Crit. Rev. Food Sci. Nutr. 2011, 51, 285. [Crossref]
  » Crossref
• ³⁸ Chaud, M.; Souto, E. B.; Zielinska, A.; Severino, P.; Batain, F.; Oliveira-Junior, J.; Alves, T.; Toxics 2021, 9, 131. [Crossref]
  » Crossref
• ³⁹ Pavoni, L.; Pavela, R.; Cespi, M.; Bonacucina, G.; Maggi, F.; Zeni, V.; Canale, A.; Lucchi, A.; Bruschi, F.; Benelli, G.; Nanomaterials 2019, 9, 1285. [Crossref]
  » Crossref
• ⁴⁰ Peres, M. C.; de Souza Costa, G. C.; dos Reis, L. E. L.; da Silva, L. D.; Peixoto, M. F.; Alves, C. C. F.; Forim, M. R.; Quintela, E. D.; Araújo, W. L.; de Melo Cazal, C.; J. Pest Sci. 2020, 93, 807. [Crossref]
  » Crossref
• ⁴¹ Solans, C.; Sole, I.; Curr. Opin. Colloid Interface Sci. 2012, 17, 246. [Crossref]
  » Crossref
• ⁴² de Oliveira, A. C.; Sá, I. S.; Mesquita, R. S.; Pereira, B. L.; Pocrifka, L. A.; de Souza, T. P.; Rodriguez Amado, J. R.; Azevedo, S. G.; Sanches, E. A.; Nunomura, S. M.; Rev. Bras. Farmacogn. 2020, 30, 667. [Crossref]
  » Crossref
• ⁴³ Roy, A.; Guha, P.; J. Food Process. Preserv. 2018, 6, e13617. [Crossref]
  » Crossref
• ⁴⁴ Vinh, T. D. T.; Hien, M. T. L.; Dao, A. T. D.; Food Sci. Nutr. 2020, 4, 7411752. [Crossref]
  » Crossref
• ⁴⁵ Toxicidade do óleo essencial de Piper tuberculatum Jacq. (Piperaceae) sobre adultos de Cerotoma arcuatus (Oliv.) (Coleoptera: Chrysomelidae), https://www.alice.cnptia.embrapa.br/alice/handle/doc/921826, accessed in February 2025
  » https://www.alice.cnptia.embrapa.br/alice/handle/doc/921826
• ⁴⁶ Toxicidade do óleo essencial de Piper Tuberculatum JACQ. (Piperaceae) sobre ninfas de primeiro instar de Crinocerus Sanctus (FABR.) (Hemiptera, Coreidae), https://www.alice.cnptia.embrapa.br/alice/handle/doc/867994, accessed in February 2025
  » https://www.alice.cnptia.embrapa.br/alice/handle/doc/867994
• ⁴⁷ Krinski, D.; Foerster, L. A.; Cienc. Agrotecnol. 2016, 40, 676. [Crossref]
  » Crossref
• ⁴⁸ Lee, B. H.; Choi, W.-S.; Lee, S.-E.; Park, B.-S.; Crop Prot. 2001, 20, 317. [Crossref]
  » Crossref
• ⁴⁹ Pavela, R.; Benelli, G.; Trends Plant Sci. 2016, 12, 10001007. [Crossref]
  » Crossref

Edited by

• Editor handled this article:
  Hector Henrique F. Koolen (Associate)

Publication Dates

• Publication in this collection
  28 Feb 2025
• Date of issue
  2025

History

• Received
  18 July 2024
• Accepted
  10 Feb 2025