The aim of this study was to develop a chemically stable plant following a pre-domestication process and the valorization of its essential oil for the production of biopesticides. This study was conducted during four growing seasons to give a pre-domesticated population (SAMO-0). The resulting pre-domesticated population increased the hydrodistilled oil yield (average 0.45%) and maintained a stable yield of dry material (44.5%). The plant material was submitted to pilot plant scale steam distillation under three pressures (0.5, 1.0 and 1.5 bar) with lower yield (average 0.15%). The essential oil showed a carvacrol /p-cymene chemotype. The pre-domestication process increased beta-myrcene, alpha- and gamma-terpinene, p-cymene, thymol and beta-bisabolene: and decreased alpha-thujene and carvacrol. The steam distillation increased the oil content in alpha-thujene, alpha-pinene, alpha-terpinene, p-cymene and trans-caryophyllene, and decreased bomeol, thymoquinone thymol and beta-bisabolene. Pressure increased alpha-terpinene, thymol and carvacrol Additionally, the study of the biocidal effects (against the insect pests Spodoptera littoralis, Myzus persicae and Leptinotarsa decemlineata and the phytopathogenic nematode Meloydogine javanica) of the EOs showed that overall, the most active oils were the hydrodistilled (to all insect species), followed by the steam distilled oils with higher carvacrol and thymol content (pressures of 1.5 and 1.0 bar). Carvacrol and thymol were responsible for the activity of these oils on M. persicae, and L. decemlineata but only partially on S. littoralis. The steam distilled oils showed strong nematicidal activity against M. javanica that could be partially explained by their content in active carvacrol and thymol.

The insecticidal and acaricidal effects of the essential oils (EO) from two populations of Geranium macrorrhizum L (ornamental variety Bevan, VB and wild population from Hungary, GH) have been studied against several insect pests of economic importance in agriculture (Spodoptera littoralis, Myzus persicae and Rhopalosiphum padi), and the hard tick Hyalomma lusitanicum. The oil from the wild population (GH) was characterized by its content in β-elemenone (30.53%), thymol (18.52%) and germacrone (15.54%), while the oil from the commercial variety (VB) had linalool (26.45%) and linalyl acetate (25.11%) as major components. S.litoralis and R. padi were significantly affected by the wild population oil (GH-EO). The oil from the commercial variety VB was less effective against insects. The ixodicidal effects of these oils were strong, being the wild population (GH-EO) the most effective. To correlate the composition of these oils with their activity, the essential oil major components have been isolated (germacrone and β-elemenone) and tested (linalool, thymol, germacrone and β-elemenone). The insect antifeedant and ixodicidal effects of thymol and germacrone explained the bioactivity of the GH essential oil. Therefore, the wild variety of G. macrorrhizum has been selected for domestication given its potential as a source of bioactive extracts and metabolites.

The nematicidal activity of hydrolate by-products from the semi industrial vapor-pressure essential oil extraction of selected aromatic plant species (commercial: Lavandula × intermedia Emeric ex Loisel. var. super, Thymus vulgaris L., T. zygis Loefl ex L. and experimentally pre-domesticated: L. luisieri (Rozeira) Rivas-Martínez) was investigated against the root-knot nematode Meloidogyne javanica by in vitro and in vivo bioassays. Liquid-liquid extraction of hydrolates yielded the corresponding aqueous and organic fractions which were biological and chemically studied. Hydrolates from L. × intermedia var. super, L. luisieri , T. vulgaris , and T. zygis showed strong in vitro nematicidal effects against M. javanica (J2 mortality and suppression of egg hatching). In the case of the Thymus species, the active components were found in the organic fraction, characterized by thymol as major component. Conversely, the nematicidal activity of L. × intermedia var. super and L. luisieri remained in the corresponding aqueous fractions. In vivo tests on tomato seedlings at sublethal doses of the hydrolates/organic fractions induced a significant reduction of nematode infectivity. In pot experiments, all hydrolates tested on tomato plants significantly affect the infection frequency and reproduction rate of the nematode population. This study demonstrates that L. × intermedia var. super, L. luisieri , T. vulgaris , and T. zygis hydrolates could be an exploitable source of potential waste protection products on root-knot nematodes.

Several extracts from pre-domesticated Lavandula luisieri (Rozeira) Riv-Mart. (Lamiaceae) (essential oil, EO; hexane, Hx and the organic fraction of the residual hydrolate, WRO) showed larvicidal effects against the hard tick Hyalomma lusitanicum, being WRO the most active. Their content in the necrodane-type monoterpenes (2,2,3,4-tetramethyl-5-oxocyclopent-3-en-1-yl)-methyl acetate (2), 5-hydroxymethyl-2,3,4,4-tetramethylcyclopent-2-en-1-one (1) and 2-(hydroxymethyl)-3,4,4-trimethyl-5-methylenecyclopent-2-en-1-one (5) correlated with these effects. Among the isolated compounds, 3,3,4,5-tetramethyl-2H-pyran-2,6(3H)-dione (7) and (2,2,3,4-tetramethyl-5-oxocyclopent-3-en-1-yl)-methyl acetate (2) were the most potent ones, with activity levels within the range of the positive control thymol. These compounds represent a new class of ixodicidal agents.

Fourteen essential oils (EOs) from selected live germplasm of medicinal plants have been tested for their antitrypanosomal and cytotoxic activity. These plants have been domesticated and maintained under experimental cultivation. Their EOs were tested on epimastigote forms of Trypanosoma cruzi strain Y and human lung fibroblasts LC5 cell line, along with the major components of the active oils, both separately and in binary combinations. Mentha rotundifolia, Thymus zygis, T. vulgaris and Hyssopus officinalis were the most active EOs against T. cruzi. Among the main components of these EOs (1-8-cineole, thymol, p-cymene, piperitenone oxide, β-pinene, γ-terpinene, carvacrol and linalool), the most active against the parasite and less toxic to human cells was thymol. In general, the activity of the main components did not exceed that of their origin EO, and the study of the activity of these compounds in combination indicates the existence of antagonistic and synergistic effects depending on the concentration tested.

Several preparations were obtained from the aerial parts of predomesticated Lavandula luisieri, including the essential oil and ethanolic, hexane, and ethyl acetate extractives. Additionally, pilot plant vapor pressure extraction was carried out at a pressure range of 0.5-1.0 bar to give a vapor pressure oil and an aqueous residue. A chemical study of the hexane extract led to the isolation of six necrodane derivatives (1, 2, and 4-7), with four of these (1, 2, 5, and 7) being new, as well as camphor, a cadinane sesquiterpene (9), tormentic acid, and ursolic acid. The EtOAc and EtOH extracts contained a mixture of phenolic compounds with rosmarinic acid being the major component. Workup of the aqueous residue resulted in the isolation of the necrodane 3 and (1R*,2S*,4R*)-p-menth-5-ene-1,2,8-triol (8), both new natural compounds. The structures of the new compounds were established based on their spectroscopic data. The phytotoxic and nematicidal activities of these compounds were evaluated.

The objective of this study was the characterization of nematicidal activity and valorization of hydrolate obtained as a byproduct of the semi industrial vapor-pressure essential oil extraction of a domesticated Artemisia absinthium population (Teruel, Spain), in comparison with a population (Sierra Nevada, Spain) undergoing the domestication process. Hydrolates from three crops of these two populations showed strong in vitro nematicidal effects against the root knot nematode Meloidogyne javanica and similar chemical profiles. The hydrolate of the A. absinthium population from Teruel strongly suppressed nematode egg hatching (>95%) after 5 days of incubation. In vivo tests on tomato seedlings showed a significant reduction in the of root penetration rate of M. javanica J2 treated with a hydrolate sublethal concentration (33%). In pot experiments, the hydrolate of A. absinthium tested on tomato plants significantly affected the nematode population, the infection frequency and the reproduction rate. The extraction of the organic fraction of the hydrolate gave a nematicidal extract and an inactive aqueous residue. The chemical profile of the active organic extract showed the presence of (5Z)-2,6-dimethylocta5,7-diene-2,3-diol, previously described in the hydrodistilled essential oil of this A. absinthium population, among other compounds. This study demonstrates that the organic fraction of the A. absinthium hydrolate is a potential root-nematode control agent. Therefore, the hydrolate byproduct of the semi industrial vapor-pressure essential oil extraction could be a source of new nematicidal products. (C) 2016 Elsevier Ltd. All rights reserved.

The objective of this study was the characterization and valorization of the essential oil from a domesticated Artemisia absinthium population (Teruel, Spain), and its comparison with another one (Sierra Nevada, Spain) undergoing the domestication process. These populations are being experimentally cultivated in the same field since 2008. We studied their biomass and essential oil production (Clevenger hydrodistillation, HD and semi-industrial vapor-pressure, VP). The domesticated population showed lower chemical variation and higher biomass and essential oil yields, allowing for the registration of a new plant variety. The observed variations in oil composition (HD, VP) between the two populations were mostly quantitative. The oils were characterized by the presence of cis-epoxyocimene, (−)-cis-chrysanthenol, chrysanthenyl acetate, linalool and trans-caryophyllene. The insect antifeedant (Leptinotarsa decemlineata, Spodoptera littoralis, Myzus persicae and Rhopalosiphum padi) and antifungal (Fusarium spp. and Botrytis cinerea) effects of their oils were also tested. All VP extracts showed strong antifungal effects and the active antifungal compounds have been identified.

Abstract Essential oils (EOs) obtained from two crops and populations of thujone-free cultivated Artemisia absinthium were tested against two nematode models, the mammalian parasite Trichinella spiralis, and the plant parasitic root knot nematode Meloidogyne javanica. The EOs were characterized by the presence of (Z)-epoxyocimene and chrysanthenol as major components and showed time and population dependent quantitative and qualitative variations in composition. The EOs showed a strong ex vivo activity against the L1 larvae of the nematode Trichinella spiralis with a reduction of infectivity between 72 and 100% at a dose range of 0.5–1 mg/ml in absence of cytotoxicity against mammalian cells. Moreover, the in vivo activity of the EO against T. spiralis showed a 66% reduction of intestinal adults. However, these oils were not effective against M. javanica.

Artemisia absinthium is an aromatic and medicinal plant of ethnopharmacological interest and it has been widely studied. The use of A. absinthium based on the collection of wild populations can result in variable compositions of the extracts and essential oils (EOs). The aim of this paper is the identification of the active components of the vapour pressure (VP) EO from a selected and cultivated A. absinthium Spanish population (T2-11) against two parasitic protozoa with different metabolic pathways: Trypanosoma cruzi and Trichomonas vaginalis. VP showed activity on both parasites at the highest concentrations. The chromatographic fractionation of the VP T2-11 resulted in nine fractions (VLC1-9). The chemical composition of the fractions and the antiparasitic effects of fractions and their main compounds suggest that the activity of the VP is related with the presence of trans-caryophyllene and di-hydrochamazulene (main components of fractions VLC1 and VLC2 respectively). Additionally, the cytotoxicity of VP and fractions has been tested on several tumour and no tumour human cell lines. Fractions VLC1 and VLC2 were not cytotoxic against the nontumoural cell line HS5, suggesting selective antiparasitic activity for these two fractions. The VP and fractions inhibited the growth of human tumour cell lines in a dose-dependent manner.

The objective of this study was the preparation of supercritical (SCE), conventional hydrodistilled (HDE) and organic solvent (OSE) extracts from two Iberian Lavandula luisieri populations (A and B) to study their chemical composition and their insect antifeedant properties against Spodoptera littoralis and Myzus persicae. HDE and SCE extracts had different chemical compositions and insect antifeedant effects, extracts from population A being more effective. Compounds 5-hydroxymethyl-2,3,4,4-tetramethylcyclopent-2-en-1-one (6) and (2,2,3,4-tetramethyl-5-oxocyclopent-3-en-1-yl)-methyl acetate (7)were the major components of the SCE fractions of both populations. The supercritical technique (SFE) improved the extraction of 6, 7, oxo-cadinol (8), hexadecanoic acid and several not identified compounds with respect to HDE. SCE extracts also exhibited increased antifeedant effects against S. littoralis and the aphid M. persicae (A-SCE). The biological effects of the active extracts cannot be accounted for those of their individual major components, suggesting additive or synergistic effects.

The aim of this study is to develop a method and to determine by means of isothermal microcalorimetry the activity of supercritical extracts obtained from growth-controlled wormwood (Artemisia absinthium L.) over Spodoptera littoralis L. (S. littoralis), a polyphagous pest of the Mediterranean crops. In order to achieve this goal, a three-step microcalorimetric method has been developed to insure the quality of the measurements and the validity of the results. Once optimized, different extracts of wormwood obtained by means of supercritical fluid extraction and by traditional methods have been used to investigate their effects over S. littoralis. The microcalorimetric method serves as a tool to complement other bioassays found in the literature. Several extracts from supercritical extractions with solvent density ranging from 485.5 to 819.5 kg m−3 have been tested. The influence of an entrainer addition to the supercritical fluid has also been investigated. The traditional extracts were obtained by hydrodistillation and organic Soxhlet extraction. The supercritical extracts were more efficient against S. littoralis than the traditionally obtained extracts.

The objective of this study was the valorization of the essential oils from Spanish Artemisia absinthium domesticated plants from Teruel and Sierra Nevada (Spain). These populations were experimentally cultivated in the field and under controlled conditions. The insect antifeedant properties of their essential oils collected yearly from two locations were tested against Spodoptera littoralis, Myzus persicae and Rhopalosiphum padi. Additionally we studied their phytotoxic, antifungal and antiparasitic effects. The oils from cultivated A. absinthium were characterized by the presence of cis-epoxyocimene, chrysanthenol, and chrysanthenyl acetate. The variations observed in oil composition were mostly quantitative but also qualitative. (Z)-2,6-Dimethyl-5,7-octadien-2,3-diol has been isolated and identified by NMR. Among the oil samples, these rich in cis-epoxyocimene and sesquiterpenes were the most active ones against S. littoralis. (Z)-2,6-Dimethyl-5,7-octadien-2,3-diol showed moderate activity against S. littoralis. The strongest antifeedant effects were found for commercial A. absinthium oil samples rich in thujones and sabinyl acetate. F. oxysporum and F. solani were affected by oils from cultivated A. absinthium and commercial oil samples. Oils from cultivated A. absinthium showed antiparasitic effects against Leishmania infantum and Trypanosoma cruzi with better results than the commercial samples.

Chagas disease is an important vector-borne disease problem in South America, especially in rural areas where inhabitants are in contact with the reduviid insects that transmit the parasite Trypanosoma cruzi. Today, the main means of interrupting transmission of T. cruzi is to control the vector. Therefore, studies of new agents with activity against these vectors have a priority interest. This review covers recent studies on essential oils from plants that have demonstrated moderate to high activity against the main vectors of Chagas disease. Further, we investigate the constituents of essential oils of plants of the genera Mentha, Thymus, Satureja and Artemisia and their activity on Rhodnius prolixus using an excito-repellency test.

The objective of this study was the valorization of Spanish Artemisia absinthium populations from Teruel (Aragón) and Sierra Nevada (Granada). These populations were experimentally cultivated in the field and under controlled conditions. Three major components were isolated from a two year-old population obtained from the Teruel population cultivated in Ejea-Zaragoza in 2003, and identified by NMR experiments as the sequiterpene lactone hydroxypelenolide (I) and the flavones artemetin (II) and casticin (III). The I–III content of the plant extracts was analyzed by HPLC-DAD. The insect antifeedant properties of plant extracts from different years and crops were tested against Spodoptera littoralis, Myzus persicae and Rhopalosiphum padi. Additionally we studied their antioxidant, phytotoxic and antiparasitic effects.

Plant parasitic nematodes are the most destructive group of plant pathogens worldwide and their control is extremely challenging. Plant Essential oils (EOs) and their constituents have a great potential in nematode control since they can be developed for use as nematicides themselves or can serve as model compounds for the development of derivatives with enhanced activity. This study reviews the plant EOs evaluated as potential nematicides and their toxic effects against pinewood nematode (Bursaphelenchus xylophilus) and root-knot nematodes (Meloidogyne spp.). Additionally, the nematicidal activity to M. javanica of several EOs from Spanish aromatic plants and their components is described.

The chemical composition and biological activity of wild Lavandula luisieri (Rozeira) Rivas-Martínez populations from the south Beira interior region in central eastern Portugal are described. The essential oils of these four populations have been analyzed by GC–MS, and their antifeedant effects evaluated against Spodoptera littoralis, Myzus persicae and Rhopalosiphum padi. Relative concentration data (% values) was subjected to principal component and cluster analysis. Relative concentration data and antifeedant effects were subjected to a stepwise multiple regression analysis. Additionally, one of these populations has been adapted to experimental field cultivation.

A comparison between traditional extraction techniques (hydrodistillation and organic solvent extraction) and supercritical fluid extraction was made for two different populations and crops of Artemisia absinthium L., cultivated in the field and aeroponically. The composition of the extracts, volatile and non volatile oils, was analyzed by GC–MS and HPLC–DAD, respectively. The antifeedant and phytotoxic activity of the extracts was tested on insect pests (Spodoptera littoralis, Myzus persicae and Rhopalosiphum padi) and plants (Lactuca sativa and Lolium perenne). The supercritical extracts exhibited stronger antifeedant effects than the traditional ones (up to 8 times more active) with moderate selective phytotoxic effects on L. perenne root growth (<50% inhibition).

Supercritical fluid extraction (SFE) of the volatile oil from Thymus vulgaris L. aerial flowering parts was performed under different conditions of pressure, temperature, mean particle size and CO(2) flow rate and the correspondent yield and composition were compared with those of the essential oil isolated by hydrodistillation (HD). Both the oils were analyzed by GC and GC-MS and 52 components were identified. The main volatile components obtained were p-cymene (10.0-42.6% for SFE and 28.9-34.8% for HD), gamma-terpinene (0.8-6.9% for SFE and 5.1-7.0% for HD), linalool (2.3-5.3% for SFE and 2.8-3.1% for HD), thymol (19.5-40.8% for SFE and 35.4-41.6% for HD), and carvacrol (1.4-3.1% for SFE and 2.6-3.1% for HD). The main difference was found to be the relative percentage of thymoquinone (not found in the essential oil) and carvacryl methyl ether (1.0-1.2% for HD versus t-0.4 for SFE) which can explain the higher antioxidant activity, assessed by Rancimat test, of the SFE volatiles when compared with HD. Thymoquinone is considered a strong antioxidant compound.