© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/)., In this work, we have investigated the accumulation of ryanoids in different plant parts (leaves, stems and roots) of aeroponically grown Persea indica cloned trees (one-year-old cloned individuals) and a selected mature, wild tree. We tested the insect antifeedant (against Spodoptera littoralis, Myzus persicae and Rhopalosiphum padi) and nematicidal (against Meloidogyne javanica) effects of ethanolic extracts from these different plant parts. The HPLC-MS analysis of P. indica extracts showed that mature tree (wild) leaves had two times more chemical diversity than stems. Aeroponic plants showed fewer differences in chemical diversity between leaves and stems, with the lowest diversity found in the roots. Ryanodane epiryanodol (1) was present in all the plant parts, with the mature stems (wild) containing the highest amount. The aeroponic stems also accumulated ryanoids including 1, cinnzeylanol (2) and cinnzeylanone (4). The insect Spodoptera littoralis was strongly affected by the stem extracts, while leaf extracts were moderately active. Based on predicted vs. real antifeedant values, we concluded that the ryanoid content (1 or a combination of 2, 4 and 1) explained the antifeedant effects of the stem extracts, while additional components contributed to the activity of the leaf extracts. Therefore, careful individual selection of P. indica seedlings should be carried out prior to proceeding with aeroponic cultivation in order to obtain ryanodane-rich stem or leaf extracts with strong antifeedant effects on S. littoralis., This work has been supported by grant MINISTERIO DE ECONOMÍA Y COMPETITIVIDAD, PID2019-106222RB-C31/SRA (State Research Agency, 10.13039/501100011033)., Peer reviewed

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Departamento de Reproducción Animal​, This work has demonstrated the ixodicidal and insect antifeedant effects of essential oils from 14 experimentally cultivated aromatic plants. The strong ixodicidal and antifeedant oils corresponded to Thymus zygis, Thymus vulgaris, Satureja montana, Oreganum virens, and Mentha suaveolens. The moderately active oils were from Lavandula angustifolia, Mentha piperita, Mentha spicata, Artemisa herba-alba, and Rosmarinus officinalis. The most effective larvicidal and antifeedant compounds were piperitenone oxide, carvacrol, piperitenone, and thymol, explaining the effects of the most active essential oils. The rest of the tested compounds were not ixodicidal or antifeedant. Therefore, the activity of moderately active oils cannot be explained by their main components (linalyl acetate, linalool, menthone, menthol, limonene, camphor, 1,8-cineole, p-cymene, α-pìnene, and carvone), suggesting synergistic effects. Considering the ixodicidal and antifeedant effects of these extracts, the plants have been ranked in relation to Thymus vulgare, a commercial biopesticide ingredient, for their potential as botanical pesticides. T. zygis, S. montana, and M. suaveolens ranked over T. vulgaris as ixodicidal agents and S. montana as insecticidal. Therefore, we propose the plant populations of S. montana, T. zygis, and M. suaveolens tested here for further development as biopesticide ingredients., This work has been supported by grants from ERASMUS+ no. 2018-1-FR01-KA202-047892, Biocontrol E Training (BET), and ERASMUS+ 600873-EPP-1-2018-1-ES-EPPKA2. European Hub on New Challenges in the Field of Essential Oils (EOHUB), PID2019-106222RB-C31, MCI, Spain, RTA 2014-00080-00-00, Spain, and the R+D contracts CSIC-Villamagna Estudio Biodirigido de Extractos Ixodicidas and UCM-Villamagna SL Investigación en torno al control de las garrapatas y al estado sanitario de la fauna silvestre., Peer reviewed

© 2022 Bautista-Sopelana, Bolívar, Gómez-Muñoz, Martínez-Díaz, Andrés, Alonso, Bravo and González-Coloma. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms., Plants are not only used as energy and nutrient resources for herbivores. Plants can be ingested because of their activity against host parasites and other pathogens. This so-called medicinal role of plants is well reported in ethnopharmacology and under-reported in wild animals. More studies on wild animals are needed because any plant in the world contains bioactive compounds, and probably all plants, no matter how toxic they are, experience herbivory. For example, we tested the activity of extracts and essential oils from Papaver rhoeas and Echium plantagineum against a selection of laboratory pathogens because Great bustards Otis tarda preferred these plants during the mating season, with male fecal droppings showing a higher frequency of P. rhoeas particles than the fecal droppings of females. We hypothesized that P. rhoeas could be helpful for males in the mating season if any part of this plant harbors bioactivity against parasites and other pathogens. Males¿ immune system is weakened during the mating season because of their investment in secondary sexual characters and sexual display. As a first exploration of the bioactivity of these plants, we evaluated extracts of both plants against a sample of laboratory models, including a flagellated protozoon (Trichomonas gallinae), a nematode (Meloidogyne javanica) and a fungus (Aspergillus niger). Non-polar and polar extracts of the aerial parts of P. rhoeas, especially the extracts of flowers and capsules, and the extracts of leaves and flowers of E. plantagineum showed activity against nematodes and trichomonads. The bioactivity of plants against parasites could explain the foraging behavior of stressed animals. The chemical communication underpinning the capacity of fauna to recognize those plants is far less known., This research was funded by FEDER/Ministerio de Ciencia, Innovación y Universidades – Agencia Estatal Consejo Superior de Investigaciones Científicas of Spain, grants no. CGL2017-87206-P and PID2019-106222RB-C31. PB was funded with a training contract of European Social Fund and Comunidad de Madrid no. PEJ-2018-AI/AMB-10263., Peer reviewed

The chemical composition of essential oils (EOs) from ten Peruvian Piper species (Piper coruscans, Pc; P. tuberculatum, Pt; P. casapiense, Pcs; P. obliquum, Po; P. dumosum, Pd; P. anonifolium, Pa; P. reticulatum, Pr; P. soledadense, Ps; P. sancti-felicis, Psf and P. mituense, Pm) has been studied, along with their antifungal and phytotoxic activities. These EOs contained β-bisabolene/nerolidol (Pc), β-bisabolene/δ-cadinene/caryophyllene (Pt), caryophyllene oxide (Pcs), bicyclogermacrene/10-epi-Elemol (Po), bicyclogermacrene/germacrene-D/apiol (Pd), caryophyllene/germacrene-D (Pa), germacrene-D (Pr), limonene/apiol (Ps), apiol (Psf), and apiol/bicyclogermacrene (Pm) as major components, and some are described here for the first time (Ps, Pcs, Pm). A composition-based dendrogram of these Piper species showed four major groups (G1: Pc and Pt, G2: Pcs, Po, Pd, Pa, and Pr, G3: Ps, and G4: Psf and Pm). The spore germination effects (Aspergillus niger, Botrytis cinerea, and Alternaria alternate) and phytotoxicity (Lolium perenne and Lactuca sativa) of these EOs were studied. Most of these Piper essential oils showed important activity against phytopathogenic fungi (except G1), especially against B. cinerea. Similarly, most of the essential oils were phytotoxic against L. perenne (except G1), with P. sancti-felicis (G4), P. casapiense (G2), and P. reticulatum (G2) being the most effective. Caryophyllene oxide, β-caryophyllene, β-pinene, limonene, α-humulene, and apiol were evaluated against B. cinerea, with the most effective compounds being β-pinene, apiol, and limonene. This work demonstrates the species-dependent potential of essential oils from Peruvian Piper species as fungicidal and herbicidal agents., This work has been financed by grants FONDECYT-CONCYTEC basic investigation (N° 433-2019-FONDECYT), Peru and PID2019-106222RB-C31/SRA (State Research Agency, Spain 10.13039/501100011033)., Peer reviewed

In this work, we have studied the benzofurans of Pericallis echinata (aerial parts and transformed roots), P. steetzii (aerial parts and transformed roots), P. lanata (aerial parts), and P. murrayi (aerial parts and roots). This work has permitted the isolation of the new benzofurans 10-ethoxy-11-hydroxy-10,11-dihydroeuparin (10), (-)-eupachinin A ethyl ether (12), 11,15-didehydro-eupachinin A (13), 10,12-dihydroxy-11-angelyloxy-10,11-dihydroeuparin (14), 2,4-dihydroxy-5-formyl-acetophenone (15) isolated for the first time as a natural product, 11-angelyloxy-10,11-dihydroeuparin (16), and 12-angelyloxyeuparone (17), along with several known ones (1–9, 11). In addition, the incubation of the abundant component, 6-hydroxytremetone (1), with the fungus Mucor plumbeus has been studied. Benzofurans in the tremetone series (1, 1a, 2–5, 18, 18a), the euparin series (6, 7, 7a, 8–10, 14, 16), and the eupachinin-type (11, 12) were tested for antifeedant effects against the insect Spodoptera littoralis. The antifeedant compounds (1, 4, 6, 11, 12) were further tested for postingestive effects on S. littoralis larvae. The most antifeedant compounds were among the tremetone series, with 3-ethoxy-hydroxy-tremetone (4) being the strongest antifeedant. Glucosylation of 1 by its biotransformation with Mucor plumbeus gave inactive products. Among the euparin series, the dihydroxyangelate 14 was the most active, followed by euparin (6). The eupachinin-type compounds (11, 12) were both antifeedants. Compounds 4, 11, and 12 showed antifeedant effects without postingestive toxicity to orally dosed S. littoralis larvae. Euparin (6) had postingestive toxicity that was enhanced by the synergist piperonyl butoxide., This work has been supported by grant PID2019-106222RB-C31, MCI, Spain., Peer reviewed

The development of Cannabis sativa strains with high cannabidiol (CBD) and low tetrahydrocannabinol (THC) content is a growing field of research, both for medical and recreational use. However, the mechanisms behind clinical actions of cannabinoids are still under investigation, although there is growing evidence that mitochondria play an important role in many of them. Numerous studies have described that cannabinoids modulate mitochondrial activity both through activation of mitochondrial cannabinoid receptors and through direct action on other proteins such as mitochondrial complexes involved in cellular respiration. Thus, the aim of this study was to determine the actions of a panel of extracts, isolated from high-CBD varieties of Cannabis sativa, on the activity of the mitochondrial electron transport chain complex IV, cytochrome c oxidase (CCO), in order to select those with a safer profile. After demonstrating that Cannabis sativa strains could be identified by cannabinoids content, concentration–response curves were performed with a collection of extracts from strains with high-CBD and low-THC content using bovine CCO. The CCO rate was clearly modified by specific extracts of Cannabis sativa plants compared to others. Half maximal inhibitory concentrations (IC50) of extracts and the inhibitory effects evoked at 1 × 10−4 g/mL displayed a significant correlation with the THC. Therefore, the screening of extracts based on CCO activity provides a powerful and rapid methodology to identify those plants with higher mitochondrial toxicity or even mito-protective actions., This work has been financed by grant PID2019-106222RB-C31/SRA, the Spanish Government (Ayudas para contratos para la formación de doctores en empresas (Doctorados Industriales) 2020, DIN2020-011349), and Instituto de Salud Carlos III (Grant number PI19/00170).

Kinetoplastida is a group of flagellated protozoa characterized by the presence of a kinetoplast, a structure which is part of a large mitochondria and contains DNA. Parasites of this group include genera such as Leishmania, that cause disease in humans and animals, and Phytomonas, that are capable of infecting plants. Due to the lack of treatments, the low efficacy, or the high toxicity of the employed therapeutic agents there is a need to seek potential alternative treatments. In the present work, the antiparasitic activity on Leishmania infantum and Phytomonas davidi of 23 essential oils (EOs) from plants of the Lamiaceae and Asteraceae families, extracted by hydrodistillation (HD) at laboratory scale and steam distillation (SD) in a pilot plant, were evaluated. The chemical compositions of the EOs were determined by gas chromatography-mass spectrometry. Additionally, the cytotoxic activity on mammalian cells of the major components from the most active EOs was evaluated, and their anti-Phytomonas and anti-Leishmania effects analyzed. L. infantum was more sensitive to the EOs than P. davidi. The EOs with the best anti-kinetoplastid activity were S. montana, T. vulgaris, M. suaveolens, and L. luisieri. Steam distillation increased the linalyl acetate, β-caryophyllene, and trans-α-necrodyl acetate contents of the EOs, and decreased the amount of borneol and 1,8 cineol. The major active components of the EOs were tested, with thymol being the strongest anti-Phytomonas compound followed by carvacrol. Our study identified potential treatments against kinetoplastids., This work has been partially financed by Grants PR108/20-08 (Santander-UCM), PID2020-114207RB-I00 and PID2019-106222RB-C31/SRA (Spanish Ministry of Science and Innovation)., Peer reviewed

© 2023 Mbula, Andres, Kitete, Kasiama, Tshilanda, Ngbolua, Tshibangu, Onautshu, González-Coloma and Mpiana. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms., The chemical composition, insect antifeedant, in vtro/in vivo nematicidal activity, phytotoxicity, and in silico nematicidal activity of the essential oil (EO) of the African medicinal plant Drypetes gossweileri were studied. Chemical analysis using GC/MS indicated that benzyl isothiocyanate (96.23%) was the major compound, followed by benzyl cyanide (1.38%). The biocidal effects of this oil were tested against insect pests and root-knot nematodes. All the insect species tested were significantly affected by the oil according to their feeding adaptations (Spodoptera littoralis and Myzus persicae were less affected than Rhopalosiphum padi) with efficient doses (EC50) of 29.4 8.3 μg/cm2, 14.744 8.3 μg/cm2, and 8.3 μg/cm2, respectively. The oil was highly effective against juveniles J2 of the nematode Meloidogyne javanica, with LC50–LC90 values of 0.007 mg/mL–0.0113 mg/mL. D. gossweileri EO at minimum lethal concentrations (MLC) and below strongly inhibited egg hatching in vitro, whereas soil treatment caused a strong suppression of nematode population, infection frequency, and multiplication rate. The EO inhibited ryegrass (Lolium perenne) germination at 0.4 mg/mL, while at 0.1 mg/mL, its effects on germination, root and leaf growth were moderate (32.4%, 8.4%, and 18.3%, respectively). The tomato (Solanum lycopersicum) germination was not affected by the EO, but the root growth was reduced (56% at 0.1 mg/mL) at a dose 10 times higher than the LD50 calculated for M. javanica J2 mortality. Molecular docking of the nematicidal effects of the oil using PyRx revealed a strong interaction between potassium chloride transporting KCC3 (PDB ID: 7D90) and benzyl cyanide at a distance of 2.20 A° with GLN C:350, followed by benzyl isothiocyanate at a distance of 2.78 A° with ARG B:294. The in vivo nematicidal effects of D. gossweileri EO on M. javanica penetration and reproduction in tomato roots further support the potential of this EO as a nematicidal agent with insect antifeedant effects, which could be used by local farmers for crop protection., This study was partially funded by Grant PID2019-106222RB-C31/SRA (Agencia Española de Investigación AEI, 10.13039/501100011033)., Peer reviewed

© The Author(s) 2024. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/., The endophytic fungus Aspergillus sp. SPH2 was isolated from the stems of the endemic plant Bethencourtia palmensis and its extracts were found to have strong fungicidal effects against Botrytis cinerea and ixodicidal effects against Hyalomma lusitanicum at different fermentation times. In this study, the fungus was grown using three different culture media and two methodologies, Microparticulate Enhancement Cultivation (MPEC) and Semi-Solid-State Fermentation (Semi-SSF), to increase the production of secondary metabolites during submerged fermentation. The addition of an inert support to the culture medium (Semi-SSF) resulted in a significant increase in the extract production. However, when talcum powder was added to different culture media, unexpected results were observed, with a decrease in the production of the biocompounds of interest. Metabolomic analyses showed that the production of aspergillic, neoaspergillic, and neohydroxyaspergillic acids peaked in the first few days of fermentation, with notable differences observed among the methodologies and culture media. Mellein production was particularly affected by the addition of an inert support to the culture medium. These results highlight the importance of surface properties and morphology of spores and mycelia during fermentation by this fungal species., This work was supported by a grant MINISTERO DE ECONOMÍA Y COMPETITIVIDAD, PID2019-106222RB-C31/SRA (State Research Agency, 10.13039/501100011033) and ERASMUS+ Scholarship granted to Nicolas Reyes Castillo., Peer reviewed

© The Author(s) 2024. This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (https://creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access page (https://us.sagepub.com/en-us/nam/open-access-at-sage).
Request permissions for this article., Objective: In this study, a series of synthetic hydroxylated (5-10) and prenylated (10-17) acetophenones, chalcones (18-20) and a flavone (21) have been synthesized and evaluated on the nematode Meloidogyne javanica. Additional phenolic compounds, including salicylic acid, isopropyl salicylate, thymol, carvacrol, cumic acid, eugenol, vanillin, and vanillic acid (22-29), were also tested. Methods: Compounds 5-21 have been synthesized by a sequence of reactions and were isolated and identified by 1H-NMR, 13C-NMR, and HRMS. The in vitro nematicidal effects of the compounds were tested on J2 juveniles. The nematicidal activity was confirmed by their egg hatching inhibition effects, tested at the minimum lethal concentration (minimum dose to give 100% J2 mortality). Results: Compounds 5-18 have been previously reported and 19-21 are described here for the first time. The 2´-hydroxylated acetophenones (5-9) showed strong nematicidal effects, being 7-9 the most active. Salicylic (22) and cumic (26) acids were effective compounds. Compounds 7, 12, 22-26, active on M. javanica J2 juveniles, were also egg hatching inhibitors. Conclusions: A C-2/C-4 dihydroxylation or C-2/C-4/C-3 trihydroxylation of the acetophenone moiety improved the activity, while introducing a hydrophilic isoprene chain (10-17) eliminated the effect, with only one prenylated acetophenone (12) being active. Among the additional phenolic compounds (22-29), these with a carboxyl group instead of the acyl side were better nematicides. These results generate molecular leads for developing new nematicidal phenolic compounds., The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work has been supported by grants ICOOP-CSIC: COOPA20324L, Ministerio de Ciencia, Tecnología e Innovación Minciencias and Universidad de Antioquia, grant number 111571249860-061-2016, contract 061-2016A and grant PID2019-106222RB-C31/SRA (Agencia Española de Investigación AEI, 10.13039/501100011033). Minciencias and Universidad de Antioquia scholarship in Doctorados Nacionales Program, 647-2014 was granted to Luis Alberto González López., Peer reviewed

© 2022 Bailén, Díaz-Castellanos, Azami-Conesa, Alonso Fernández, Martínez-Díaz, Navarro-Rocha, Gómez-Muñoz and González-Coloma. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms., Trichomonas gallinae is a flagellated protozoan that parasitizes the upper digestive tract of various bird species and causes avian trichomonosis. The emergence of resistant strains to the standard treatment, based on nitroimidazoles, increases the need to find alternative therapies. In this study, 36 essential oils (EOs) from Lamiaceae and Asteraceae plant families were tested against T. gallinae trophozoites using the 3-(4,5-dimethylthiazol-2-yl-)-2,5-dipheniltetrazolium bromide (MTT) reduction assay. Among them, EOs from distinct species of Lamiaceae, including the genera Lavandula, Salvia, Thymus, Origanum, and Satureja were the ones reporting better anti-trichomonal activity, and were selected for further analysis, including chemical composition and in vitro assays. The chemical composition of the selected EOs was determined by gas chromatography followed by mass spectrometry and 19 pure compounds were tested against the protozoa, according to their higher abundance in the active EOs. Pure compounds which displayed the highest activity against T. gallinae trophozoites, ordered by highest to lowest activity, were α and β-thujones, camphene, β-pinene, linalyl acetate, thymol, 4-terpineol, γ-terpinene, α-pinene, p-cymene, D-fenchone and β-caryophyllene. A dose dependent effect was observed in most of the EOs and pure compounds tested. The toxicity test conducted in eukaryotic cell cultures with the anti-trichomonal active pure compounds showed that β-caryophyllene, camphene, α-pinene, and β-pinene were slightly toxic for Vero cells, and the selectivity index was calculated. Based on the anti-trichomonal activity and the absence of cytotoxicity results, natural products from Lamiaceae plants could be useful as alternative therapy against avian trichomonosis, mainly those containing linalyl acetate, thymol, 4-terpinenol, γ-terpinene, p-cymene and D-fenchone., This work has been partially financed by Grants PR108/20-08 (Santander-UCM), PID2020-114207RB-I00 (Spanish Ministry of Science and innovation), ERASMUS+ European Hub on New Challenges in the Field of Essential Oils (EOHUB) and PID2019-106222RB-C31/SRA (State Research Agency, 10.13039/501100011033)., Peer reviewed

© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/)., Plant essential oils (EOs) are gaining interest as biopesticides for crop protection. EOs have been recognized as important ingredients of plant protection products including insecticidal, acaricidal, fungicidal, and nematicidal agents. Considering the growing importance of EOs as active ingredients, the domestication and cultivation of Medicinal and Aromatic Plants (MAPs) to produce chemically stable EOs contributes to species conservation, provides the sustainability of production, and decreases the variations in the active ingredients. In addition to these direct effects on plant pests and diseases, EOs can induce plant defenses (priming effects) resulting in better protection. This aspect is of relevance considering that the EU framework aims to achieve the sustainable use of new plant protection products (PPPs), and since 2020, the use of contaminant PPPs has been prohibited. In this paper, we review the most updated information on the direct plant protection effects of EOs, focusing on their modes of action against insects, fungi, and nematodes, as well as the information available on EOs with plant defense priming effects., This work was supported by grants from ERASMUS+ European Hub on New Challenges in the Field of Essential Oils (EOHUB) and PID2019-106222RB-C31/SRA (State Research Agency, 10.13039/501100011033). S.K. and S.S. were funded by an international PhD fellowship from the UPM-Algerian Government., Peer reviewed

© 2022 Soudani, Poza-Carrión, De la Cruz Gómez, González-Coloma, Andrés and Berrocal-Lobo. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms., In this work, we studied the direct and indirect plant protection effects of an Artemisia absinthium essential oil (AEO) on tomato seedlings against Fusarium oxysporum sp. oxysporum radicis lycopersici (Fol). AEO exhibited a toxic effect in vitro against Fol. Additionally, tomato seedlings germinated from seeds pretreated with AEO and grown hydroponically were protected against Fol. Plant disease symptoms, including, water and fresh weight loss, tissue necrosis, and chlorosis were less pronounced in AEO-treated seedlings. AEO also contributed to plant defenses by increasing callose deposition and the production of reactive oxygen species (ROS) on seed surfaces without affecting seed germination or plant development. The essential oil seed coating also primed a durable tomato seedling defense against the fungus at later stages of plant development. RNA-seq and metabolomic analysis performed on seedlings after 12 days showed that the AEO treatment on seeds induced transcriptomic and metabolic changes. The metabolomic analysis showed an induction of vanillic acid, coumarin, lycopene, oleamide, and an unknown metabolite of m/z 529 in the presence of Fol. The StNRPD2 gene, the second largest component of RNA polymerases IV and V directly involved in de novo cytosine methylation by RNA-directed DNA methylation (RdDM), was highly induced in the presence of AEO. The host methionine cycle (MTC) controlling trans-methylation reactions, was also altered by AEO through the high induction of S-adenosyl methionine transferases (SAMts). Our results suggest that AEO treatment could induce de novo epigenetic changes in tomato, modulating the speed and extent of its immune response to Fol. The EO-seed coating could be a new strategy to prime durable tomato resistance, compatible with other environmentally friendly biopesticides., This work was supported by the Ministry of Science, Spain, European Hub on New Challenges in the Field of Essential Oils (EOHUB), and PID2019-106222RB-C31, MCIU, Spain. SS was funded by an international Ph.D. fellow from the UPM-Algerian Government from MHESR, Decree No. 14–196. ND was funded by a Ph.D. fellow DIN2018-009928 from MCIU, Spain. CP-C was supported by an I+D+i contract, Fucovasa, Spain., Peer reviewed

© The Author(s) 2024. Creative Commons Non Commercial CC BY-NC: This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (https://creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access page (https://us.sagepub.com/en-us/nam/open-access-at-sage)., Objectives: The potential forestry use of Juniperus phoenicea and Juniperus thurifera pruning woods is studied by analyzing the composition of the woods and testing the biological activities of the corresponding components with the ultimate target of finding new biopesticides. Methods: The air-dried wood from each plant was crushed and subjected to hydrodistillation, and the residue was extracted with a Soxhlet apparatus using different solvents. The corresponding extracts were fractionated, and their composition were studied by gas chromatography-mass spectrometry and nuclear magnetic resonance techniques. The ixodicidal and antifungal activities of the different samples obtained were evaluated. Results: The fraction oxygenated of the essential oil from both J. thurifera (cedrol >60%) and J. phoenicea shows a remarkable bioactivity as antitick with EC50 values of 3.4 μg/mg and 10 μg/mg, respectively. Cedrol and methyl hinokiate, present in the hexane extract J. thurifera, show a potent antifungal effect against Aspergillus niger with EC50 values of 45.99 and 52.23 μg/mL, respectively. Conclusions: Pruning woods from these species proved to be renewable and easily accessible sources of bioactive natural products such as cedrol, thujopsene, nootkatone, and totarol., The authors disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work has been supported by grants MINISTERIO de CIENCIA e INNOVACIÓN, PID2019-106222RB-C31/SRA, PID2019-106222RB-C32/SRA (State Research Agency, 10.13039/501100011033) and Unidad Asociada UGR-CSIC Bioplaguicidas: Biotecnología, Síntesis y Diversidad química., Peer reviewed

The search for new compounds with biocidal potential was carried out, focusing on the longipinenes 1–7 from the plant species Santolina viscosa Lag. Compounds 1, 2, and 5 showed remarkable molecular diversity when treated in acidic reaction conditions. Protonic, Lewis, and heterogeneous compounds were used in the treatment. Three main models of reaction have been observed: isomerization of the double bond (8–10); rearrangements to longibornane-based skeleton (11–15) and ring-opening to himachalane-based skeleton (16–18). Secolongibornane aldehydes 23 and 24 were obtained after epoxide opening under the same reaction conditions. The elucidation of the structures of the new compounds was carried out using spectroscopic data and was supported by computational theoretical calculations of 13C NMR spectra. Additionally, high-resolution mass spectrometry and single-crystal X-ray diffraction analysis were employed for certain compounds. Natural longipinenes 4–7, methyl esters 1–3 of corresponding natural carboxylic acids and the isomerized and derivatives compounds 8–19 exhibit moderate to high insecticidal activity against R. padi and M. persicae insects. Longipinene 5 shows potent inhibition against the root growth of the plants L. perenne and L. sativa, as well as compound 2 on the leaves of L. perenne. Furthermore, significant ixocidal and nematicidal activity was found for this latter compound, This work has been supported by grants MINISTERIO de CIENCIA e INNOVACIÓN, PID2019-106222RB-C31/SRA, PID2019-106222RBC32/SRA (State Research Agency, 10.13039/501100011033) and Unidad Asociada UGR-CSIC Bioplaguicidas: Biotecnología, Síntesis y Diversidad Química., Peer reviewed

The continuous search for natural product-based biopesticides from fungi isolated from untapped sources is an effective tool. In this study, we studied a pre-selected fungal endophyte, isolate Aa22, from the medicinal plant Artemisia absinthium, along with the antifungal, insect antifeedant and nematicidal compounds present in the extract. The endophyte Aa22 was identified as Stemphylium solani by molecular analysis. The antifungal activity was tested by broth microdilution against Fusarium solani, F. oxysporum, F. moniliforme and Botrytis cinerea, the insect antifeedant by choice bioassays against Spodoptera littoralis, Myzus persicae and Rhopalosiphum padi and the in vitro mortality against the root-knot nematode Meloiydogyne javanica. The structures of bioactive compounds were determined on the basis of 1D and 2D NMR spectroscopy and mass spectrometry. The ethyl acetate extract obtained from the solid rice fermentation showed mycelial growth inhibition of fungal pathogens (EC50 0.08–0.31 mg/mL), was antifeedant to M. persicae (99%) and nematicidal (68% mortality). A bioguided fractionation led to the isolation of the new compound stempholone A (1), and the known stempholone B (2) and stemphol (3). These compounds exhibited antifeedant (EC50 0.50 mg/mL), antifungal (EC50 0.02–0.43 mg/L) and nematicidal (MLD 0.5 mg/mL) activities. The extract activities can be explained by 3 (antifungal), 1–3 (antifeedant) and 1 (nematicidal). Phytotoxicity tests on Lolium perenne and Lactuca sativa showed that the extract and 1 increased L. sativa root growth (121–130%) and 1 reduced L. perenne growth (48–49%). These results highlight the potential of the endophytic fungi Aa22 as biotechnological source of natural product-based biopesticides., This work has been supported by grant PID2019-106222RB-C31, MINECO/FEDER, Department of Science and Technology (Grant no DST/INT/SPAIN/P-3/-9), India., Peer reviewed

Endolichenic fungi represent an important ecological group of microorganisms that form associations with photobionts in the lichen thallus. These endofungi that live in and coevolve with lichens are known for synthesizing secondary metabolites with novel structures and diverse chemical skeletons making them an unexplored microbial community of great interest. As part of our search for new phytoprotectants, in this work, we studied the endolichenic fungus Xylaria sp. isolated from the lichen Hypogymnia tubulosa, which grows as an epiphyte on the bark of the endemic Canarian tree Pinus canariensis. From the extract of the liquid fermentation, we isolated two unreported piliformic derivatives, (+)-9-hydroxypiliformic acid (1) and (+)-8-hydroxypiliformic acid (2), along with four previously reported compounds, (+)-piliformic acid (3), hexylaconitic acid A anhydride (4), 2-hydroxyphenylacetic acid (5), and 4-hydroxyphenylacetic acid (6). Their structures were elucidated based on NMR and HRESIMS data. The extract and the isolated compounds were tested for their insect antifeedant (Myzus persicae, Rhopalosiphum padi, and Spodoptera littoralis), antifungal (Alternaria alternata, Botrytis cinerea, and Fusarium oxysporum), nematicidal (Meloidogyne javanica), and phytotoxic effects on mono- and dicotyledonous plant models (Lolium perenne and Lactuca sativa). Compounds 4, 5, and 6 were effective antifeedants against M. persicae and 4 was also active against R. padi. Moreover, 3 and 4 showed antifungal activity against B. cinerea and 4 was the only nematicidal. The extract had a strong phytotoxic effect on L. sativa and L. perenne growth, with compounds 3, 4, and 5 identified as the phytotoxic agents, while at low concentrations compounds 3 and 4 stimulated L. sativa root growth., This work has been supported by grant PID2019-106222RB-C31, MINECO/FEDER, Spain. Work is co-financed by the Canary Islands Agency for Research, Innovation and Information Society of the Regional Ministry of Economy, Knowledge and Employment and by the European Social Fund (ESF) Canary Islands Integrated Operational Program 2014–2020, Axis 3 Priority Theme 74 (85%)., Peer reviewed

The obtained ITS1-5.8S-ITS2 sequence data was compared with these published in the NCBI (National Center for Biotechnology Information, https://www.ncbi.nlm.nih.gov/) database using Basic Local Alignment Search Tool (nBLAST). This strain was identified by similarity with another P. capitalensis strain (GenBank accession MT649668). A sample of this fungus was deposited in CECT (Valencia, Spain) under the number 20914 in compliance with the Budapest Treaty. The datasets used in the current study are available from the corresponding author on reasonable request., In the current work, the production of dioxolanone and meroterpene type secondary metabolites under different fermentation conditions by the endophytic fungus Phyllosticta capitalensis (isolate YCC4) isolated from the leaves of the endemic plant Persea indica has been evaluated. Different techniques, such as microparticle-enhanced culture (MPEC: bentonite, talcum powder) and surface adhesion fermentation (SAF: metallic mesh and glass wool) were applied to the culture medium. Ethyl acetate extracts from different fermentations of the isolate YCC4 were analyzed for their content and revealed the following dioxolanone derivatives: metguignardic acid (1), guignardianone C (2), ethyl guignardate (3), guignardianone D (4) and phenguignardic acid methyl ester (5), and meroterpenes: guignardone A (6) and B (7), guignarenone C (8), guignarenone B (9) and guignardone I (10). Additionally, the biological activity of extracts was tested against the aphid Myzus persicae and the root-knot nematode Meloidogyne javanica. Our study revealed significant variations in the production of the target metabolites as well as notable differences in biopesticide activity influenced by the presence of inert supports. Overall, the findings indicate that glass wool (GW) is a high-performance material to improve the production of dioxolanone derivatives. These findings underscore the significance of innovative cultivation methods such MPEC and SAF as drivers to produce valuable secondary metabolites from fungal organisms., This work has been funded by grant PID2019-106222RB-C31 funded by MCIN. Jorge Rojas López-Menchero has been supported by a predoctoral contract associated to the project PID2019-106222RB-C31 funded by MCIN/AEI/https://doi.org/10.13039/501100011033 and FSE “El FSE invierte en tu futuro”., Peer reviewed

This work presents the biocidal effects and chemical compositions of two essential oils (EOs) obtained by hydrodistillation of Eucalyptus flocktoniae subsp. flocktoniae and E. flocktoniae subsp. hebes. The two subspecies studied had different chemical composition, when analysed by gas chromatography coupled to mass spectrometry with 1,8-cineole (56.98%), trans-pinocarveol (20.38%) and α-pinene (5.86%) being the major components of E. flocktoniae subsp. flocktoniae and spathulenol (25.09%), p-cymene (21.20%), 1,8-cineole (10.74%) and α-pinene (8.93%) are the major components of E. flocktoniae subsp. hebes. These oils were evaluated for their insect antifeedant, nematicidal and phytotoxic activities. The biocidal tests showed that E. flocktoniae subsp. hebes was the most active against Myzus persicae and Rhopalosiphum padi. While, E. flocktoniae subsp. flocktoniae was not antifeedant. None of the investigated EOs were active against both Spodoptera littoralis and Meloidogyne javanica root-knot nematode. Moreover, the EOs extracted from these two subspecies showed a significant phytotoxic effect., This work has been partially supported by grant PID2019-106222RB-C31/SRA (State Research Agency, Spain10.13039/501100011033)., No

Endophytic fungi produce a diverse range of bioactive secondary metabolites with potential applications in biopesticide development. This study investigates the nematicidal and antifeedant properties of ethyl acetate extracts from endophytic fungi isolated from wild Arabidopsis thaliana populations in Spain. The extracts were tested against the plant-parasitic nematode Meloidogyne javanica, and two common insect pests, Myzus persicae and Spodoptera littoralis. Nine of the 13 extracts demonstrated significant nematicidal and/or antifeedant activity, indicating their potential as biopesticides. The active extracts were derived from six genera: Alternaria (3 isolates), Dydimella (1), Dothiora (1), Pleiochaeta (1), Penicillium (1), and Fusarium (2). Five extracts exhibited nematicidal activity above 90%, with three reducing the total number of M. javanica second-stage juveniles hatched from egg masses by 22-37%. Four extracts showed strong settling inhibition (> 70%) against M. persicae, and three exhibited feeding inhibition against S. littoralis. Chemical analysis by GC-MS and LC-MS revealed a wide array of unique secondary metabolites in the active extracts, reflecting substantial chemical diversity, regardless of the fungal origin. This study highlights the potential of fungal endophytes from A. thaliana as sources of novel biopesticides, paving the way for future research focused on harnessing these fungi for biopesticide development., Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature.

S.D.-G was supported by a Margarita Salas Grant for junior doctors (RD 289/2021), funded by the Spanish Ministry of Science, Innovation and Universities (MCIN/AEI/https://doi.org/10.13039/501100011033) and the European Union – NextGenerationEU and PID2021-123697OB-I00 funded by MCIN/AEI/https://doi.org/10.13039/501100011033 and “ERDF A way of making Europe”. C.G.-S was supported by grant PEJ-2020-AI/BIO-19,580 funded by Comunidad de Madrid and is currently supported by grants PRE2022-103983 and CEX2020-000999-S-20-3 funded by MCIN/AEI/ https://doi.org/10.13039/501100011033 and “ESF Investing in your future”. S.S. research is supported by grant PID2021-123697OB-I00 funded by MCIN/AEI/https://doi.org/10.13039/501100011033 and “ERDF A way of making Europe”. S.D.-G., A.G.-C. and M.A. research was supported by grant PID2019-106222RB-C31/SRA (State Research Agency, https://doi.org/10.13039/501100011033 )., With funding form the Spanish government through the "Severa Ochoa Centre of Excellence" accreditation (CEX2020-000999-S-20-3)., Peer reviewed

Leaf essential oils (EOs) of seven Eucalyptus species planted in Southern Tunisia (E. gracilis, E. lesouefii, E. salmonophloia, E. sargentii, E. flocktoniae, E. oleosa, and E. salubris) were screened for their antifeedant, nematicidal, and phytotoxic effects as well as their chemical compositions. The yield of EOs of these species varied from 1.32% ± 0.38% to 4.92%±0.37%. Using gas chromatography mass spectrometry (GC–MS) analysis, 22 compounds were identified, representing 87.16%–96.83% of the total oil content. The main components found in all EOs were 1,8-cineole (38.71%–70.44%), α-pinene (4.40%–8.88%), p-cymene (0.95%–14.99%), pinocarvone (0.17%–3.22%), trans-pinocarveol (1.53%–13.48%), spathulenol (0.18%–5.39%), and globulol (0.22%–6.47%). The insect antifeedant, nematicidal, and phytotoxic effects of these oils against insect pests (Spodoptera littoralis, Myzus persicae, and Rhopalosiphum padi), the root-knot nematode (Meloidogyne javanica), and two plant models (Lolium perenne and Lactuca sativa) were examined. Despite the moderate insect antifeedant activity, the EOs of E. salubris, E. salmonophloia, E. lesouefii, and E. sargentii were the most effective against M. persicae. Significant nematicidal effects were observed for both E. salmonophloia and E. oleosa EOs. Moreover, all Eucalyptus EOs significantly suppressed the root and leaf growth of L. perenne than the root length of L. sativa, indicating that EOs from Eucalyptus possessed an excellent phytotoxic potential., Funding: This study was supported by grant PID2019-106222RB-C31/SRA (State Research Agency, Spain 10.13039/501100011033)., Peer reviewed

[Data availability]
The dataset(s) supporting the conclusions of this article are available in the GEO repository (Record GSE295439) in https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE295439., Phytophthora cinnamomi is a destructive phytopathogen responsible for crown rot, trunk canker, and root rot in over 5,000 plant species worldwide. Despite its significant impact, the molecular interactions between this oomycete and host plants are not well understood. This study aimed to investigate the physiological and molecular responses of Solanum lycopersicum (tomato) to P. cinnamomi infection. The initial defense response in tomato seeds included the production of reactive oxygen species and callose deposition. Screening of commercial tomato varieties revealed varying levels of susceptibility, with the cultivar Marmande exhibiting heightened vulnerability. Three days postinoculation, Marmande showed increased expression of genes associated with reactive oxygen species generation and biosynthesis pathways for phenylpropanoids and flavonoids. Additionally, 850 genes related to cell wall remodeling, including those involved in lignin biosynthesis and pectin methyl esterase inhibitors, were significantly upregulated. Seven days postinoculation, a stronger transcriptional response was observed, with activation of ethylene and jasmonic acid signaling pathways, whereas salicylic acid showed minimal activity. Metabolomic analysis of infected roots revealed elevated levels of metabolites linked to lycopene, flavonoids, and phenylpropanoids. Furthermore, infected roots exhibited a significant reduction in pectin levels, which was corroborated by in vitro assays showing zoospore-mediated pectin degradation. These results suggest that degradation of root pectin is a key mechanism facilitating zoospore invasion in susceptible tomato hosts. This study provides new insights into the molecular mechanisms underlying host–pathogen interactions and identifies potential targets for managing P. cinnamomi-induced diseases in crops., Funding: Support was provided by the Ministerio de Ciencia, Innovación y Universidades (DIN2018-009928 and PID2019-106222RB-C31), the European Agricultural Fund for Rural Development (TSA0069316), the Fundación Conde del Valle Salazar, and the Direction Générale de la Recherche Scientifique et du Développement Technologique (Decree No. 14-196).

The author(s) declare no conflict of interest., Peer reviewed

Every year, more than 50 million tons of Helianthus annuus L. are produced mainly for the sunflower oil industry. It is a plant consumed worldwide, due to its use in feeding and traditional medicine for the prevention of some cardiovascular diseases, as anti-inflammatory, antimicrobial, antioxidant and antihypertensive, among others. Sunflower has a great diversity of metabolites, being especially rich in terpene compounds with close to 400 different molecules reported, including volatile mono and sesquiterpenes. Among these terpenic components of sunflower, sesquiterpenes as helibisabonols, helianuols and heliangolides, diterpenes (ent-kaurenes or ent-trachylobanes and ent-atisarenes compounds) or apocarotenoids (ionone compounds and strigolactones) are worthy to be underlined. Many of these compounds have allelopathic activity. On the other hand, sesquiterpene lactones and diterpene compounds have strong insect antifeedant, antifungal and allelopathic activities. Additionally, the essential oil (rich in α-pinene) has antifungal properties. All these biological activities suggest that Helianthus annuus has great potential for natural crop protection. Since the main sunflower crop residue is the flower head, we have revised the plant-protection properties of flowerhead components to highlight the potential valorization of that residue for the production of biopesticides., This research has been funded by grants MINISTERIO DE ECONOMÍA Y COMPETITIVIDAD, PID2019-106222RB-C32/SRA (State Research Agency, 10.13039/501100011033), PID2019-106222RB-C31/SRA (State Research Agency, 10.13039/501100011033 and Unidad Asociada UGR-CSIC Bioplaguicidas: Biotecnología, síntesis y diversidad química., Peer reviewed

Lamiaceae and Asteraceae plant species have been widely used in Mediterranean ethnomedicine for gastrointestinal disorders. They are also known for their antioxidant, anti-inflammatory, anti-bacterial, anti-parasite, and anti-virus properties. Giardia duodenalis is the most prevalent intestinal protozoon in children and young dogs worldwide. Its zoonotic potential and frequent therapeutic failures with nitroimidazoles underscore the urgent need for alternative treatments. This study investigated the antigiardial activity of essential oils (EOs) from 22 medicinal plants belonging to Lamiaceae and Asteraceae, together with their major constituents. EO composition was determined by a metabolomic approach (GC-MS). Parasite metabolic activity was assessed using the MTT assay, and ultrastructural changes were examined by Transmission Electron Microscopy. The strongest antigiardial effects were observed with Lavandula luisieri, Thymus vulgaris, Mentha suaveolens, Satureja montana (IC50 <25), L. lanata, and T. zygis, (IC50= 27.9–71.5 µg/ml). The highest selective indexes were obtained with γ-terpinene, caryophyllene oxide, carvacrol and thymol (SI≥1.3–2.4). Synergistic interactions were detected with linalyl acetate and linalool (present in Lavandula EOs), linalyl acetate with ρ-cymene or thymol, or combinations of ρ-cymene, γ-terpinene, thymol, and carvacrol (present in Satureja EOs). Transmission Electron Microscopy revealed membranolysis, enlarged periplasmic vacuoles, and cytoplasmic loss in trophozoites exposed to γ-terpinene after 1 h. These findings provide phytotherapeutic evidence supporting essential oils from Lavandula, Mentha, Thymus, and Satureja as promising antigiardial agents. Their main components γ-terpinene, caryophyllene oxide, carvacrol and thymol could have potential applications in veterinary parasitology., This work was supported by the Spanish ministry of science, innovation and universities (Grant number PID2020–114207RB-I00, Grant number PID2019–106222RB-C31/SRA, 10.13039/501100011033)., Peer reviewed

Fungal endophytes have emerged as a promising strategy for sustainable crop management, due to their remarkable ability to synthesize a wide array of secondary metabolites with beneficial effects on plants. These microorganisms establish asymptomatic relationships within plant tissues and can produce bioactive compounds with biocidal properties. Despite promising advances, the mechanisms of symbiosis remain unclear and there is a lack of in-depth analysis on the chemical characterization of their metabolites. This review compiles and discusses the current discoveries on 180 metabolites summarized according to their biological activities (insecticidal, antifungal, phytotoxic, nematicidal and antibacterial), highlighting their potential applications as biocontrol agents for integrated crop protection strategies. As a future perspective, the use of next-generation sequencing (NGS) technologies such as metagenomics in endophytic fungi research is emphasized for its promise in elucidating complex plant–microbe interactions and guiding bioprospecting efforts to identify fungal taxa capable of producing agriculturally relevant bioactive compounds. This review aims to summarize recent advances in the field, highlighting the potential uses of endophytic compounds in plant health, disease suppression, and resistance enhancement, while also identifying current limitations and outlining future research directions to further expand this promising area., Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. This work has been supported by grant PID2019-106222RB-C31/SRA MCIN/AEI/https://doi.org/10.13039/501100011033. Jorge Rojas López-Menchero has been supported by a predoctoral contract FPI associated to the project PID2019-106222RB-C31 funded by MCIN/AEI/ 10.13039/ 501100011033 and FSE “El FSE invierte en tu futuro”. Fotyos A. Lyssaos thanks to ACIISI of the Canary Islands Government for a predoctoral contract and thesis/work is co-financed by the Canary Islands Agency for Research, Innovation and Information Society of the Regional Ministry of Economy, Knowledge and Employment and by the European Social Fund (ESF) Canary Islands Integrated Operational Program 2014–2020, Axis 3 Priority Theme 74 (85%)., Peer reviewed

Lamiaceae and Asteraceae plant species have been widely used in Mediterranean ethnomedicine for gastrointestinal disorders. They are also known for their antioxidant, anti-inflammatory, anti-bacterial, anti-parasite, and anti-virus properties. Giardia duodenalis is the most prevalent intestinal protozoon in children and young dogs worldwide. Its zoonotic potential and frequent therapeutic failures with nitroimidazoles underscore the urgent need for alternative treatments. This study investigated the antigiardial activity of essential oils (EOs) from 22 medicinal plants belonging to Lamiaceae and Asteraceae, together with their major constituents. EO composition was determined by a metabolomic approach (GC-MS). Parasite metabolic activity was assessed using the MTT assay, and ultrastructural changes were examined by Transmission Electron Microscopy. The strongest antigiardial effects were observed with Lavandula luisieri, Thymus vulgaris, Mentha suaveolens, Satureja montana (IC50 <25), L. lanata, and T. zygis, (IC50= 27.9–71.5 µg/ml). The highest selective indexes were obtained with γ-terpinene, caryophyllene oxide, carvacrol and thymol (SI≥1.3–2.4). Synergistic interactions were detected with linalyl acetate and linalool (present in Lavandula EOs), linalyl acetate with ρ-cymene or thymol, or combinations of ρ-cymene, γ-terpinene, thymol, and carvacrol (present in Satureja EOs). Transmission Electron Microscopy revealed membranolysis, enlarged periplasmic vacuoles, and cytoplasmic loss in trophozoites exposed to γ-terpinene after 1 h. These findings provide phytotherapeutic evidence supporting essential oils from Lavandula, Mentha, Thymus, and Satureja as promising antigiardial agents. Their main components γ-terpinene, caryophyllene oxide, carvacrol and thymol could have potential applications in veterinary parasitology.

Lamiaceae and Asteraceae plant species have been widely used in Mediterranean ethnomedicine for gastrointestinal disorders. They are also known for their antioxidant, anti-inflammatory, anti-bacterial, anti-parasite, and anti-virus properties. Giardia duodenalis is the most prevalent intestinal protozoon in children and young dogs worldwide. Its zoonotic potential and frequent therapeutic failures with nitroimidazoles underscore the urgent need for alternative treatments. This study investigated the antigiardial activity of essential oils (EOs) from 22 medicinal plants belonging to Lamiaceae and Asteraceae, together with their major constituents. EO composition was determined by a metabolomic approach (GC-MS). Parasite metabolic activity was assessed using the MTT assay, and ultrastructural changes were examined by Transmission Electron Microscopy. The strongest antigiardial effects were observed with Lavandula luisieri, Thymus vulgaris, Mentha suaveolens, Satureja montana (IC50 <25), L. lanata, and T. zygis, (IC50= 27.9–71.5 µg/ml). The highest selective indexes were obtained with γ-terpinene, caryophyllene oxide, carvacrol and thymol (SI≥1.3–2.4). Synergistic interactions were detected with linalyl acetate and linalool (present in Lavandula EOs), linalyl acetate with ρ-cymene or thymol, or combinations of ρ-cymene, γ-terpinene, thymol, and carvacrol (present in Satureja EOs). Transmission Electron Microscopy revealed membranolysis, enlarged periplasmic vacuoles, and cytoplasmic loss in trophozoites exposed to γ-terpinene after 1 h. These findings provide phytotherapeutic evidence supporting essential oils from Lavandula, Mentha, Thymus, and Satureja as promising antigiardial agents. Their main components γ-terpinene, caryophyllene oxide, carvacrol and thymol could have potential applications in veterinary parasitology.

Kinetoplastida is a group of flagellated protozoa characterized by the presence of a kinetoplast, a structure which is part of a large mitochondria and contains DNA. Parasites of this group include genera such as Leishmania, that cause disease in humans and animals, and Phytomonas, that are capable of infecting plants. Due to the lack of treatments, the low efficacy, or the high toxicity of the employed therapeutic agents there is a need to seek potential alternative treatments. In the present work, the antiparasitic activity on Leishmania infantum and Phytomonas davidi of 23 essential oils (EOs) from plants of the Lamiaceae and Asteraceae families, extracted by hydrodistillation (HD) at laboratory scale and steam distillation (SD) in a pilot plant, were evaluated. The chemical compositions of the EOs were determined by gas chromatography-mass spectrometry. Additionally, the cytotoxic activity on mammalian cells of the major components from the most active EOs was evaluated, and their anti-Phytomonas and anti-Leishmania effects analyzed. L. infantum was more sensitive to the EOs than P. davidi. The EOs with the best anti-kinetoplastid activity were S. montana, T. vulgaris, M. suaveolens, and L. luisieri. Steam distillation increased the linalyl acetate, β-caryophyllene, and trans-α-necrodyl acetate contents of the EOs, and decreased the amount of borneol and 1,8 cineol. The major active components of the EOs were tested, with thymol being the strongest anti-Phytomonas compound followed by carvacrol. Our study identified potential treatments against kinetoplastids.