Synergistic larvicidal and repellent effects of essential oils of three Origanum species on Rhipicephalus annulatus tick

Ticks are of great economic importance worldwide, both because they represent major obstacles to livestock productivity and because of their ability to transmit diseases to humans and animals. Although synthetic acaricides are the most common method for tick control, their overuse has led to the development of resistance as well as unacceptable residual levels in animal products and in the environment in general. There is therefore an urgent need to identify alternative treatments. Among such alternative approaches for tick control is plant essential oil (EO) therapy. In the present study, we investigated the synergistic effect of EOs of three oregano species—Origanum onites, O. majorana and O. minutiflorum—against Rhipicephalus annulatus larvae. Gas chromatography–mass spectrometry profiles of the three EOs revealed that carvacrol was their major component, with a concentration of 86.2% in O. majorana, 79.1% in O. minutiflorum and 77.4% in O. onites. The results of larvicidal assays revealed that the doses that lead to the death of 50% of the ticks (LC₅₀) were 22.99, 25.08 and 27.06 µL/mL for O. majorana, O. minutiflorum and O. onites EOs, respectively, whereas the doses that lead to the death of 99% (LC₉₉) were 41.26, 43.62 and 48.96 µL/mL. In addition, the LC₅₀ and LC₉₉ of the three oils combined was lower (viz., 4.01 and 6.97 µL/mL) than that of each oil alone. The tested EOs were also able to repel larvae of R. annulatus to varying degrees, with O. onites oil exhibiting the greatest repellent effect, as shown by the lowest RC₅₀ dose, followed by O. minutiflorum and O. majorana. Interestingly, this means that the oil that was least effective in killing the larvae was the most effective in repelling them. The calculated synergistic factor of any combination was higher than 1 which means that combinations have a synergistic effect. In conclusion, the combination of all three oils showed higher toxic and repellent activities than either oil separately or combinations of any two oils, suggesting synergistic effects with low doses. Further studies including field trials and the establishment of the mode of action and side effects are urgently needed to expand on these findings, and other tick stages such as adults should also be tested.

     

Chemical Composition and Antimicrobial Activity of Origanum libanoticum,Origanum ehrenbergii,and Origanum syriacum Growing Wild in Lebanon

The essential oils (EOs) of the aerial parts of Origanum libanoticum and Origanum ehrenbergii, endemic to Lebanon, and Origanum syriacum, endemic to the Levantine, were obtained by distillation with a Clevenger apparatus. GC and GC/MS allowed identification of 96.4%, 93.5%, and 95.2% of their constituents, respectively. Carvacrol was the major component of both O. syriacum EO (79%) and O. ehrenbergii EO (60.8%). This compound was absent in O. libanoticum EO and the major compounds were β‐caryophyllene (26.8%), caryophyllene oxide (22.6%), and germacrene D (17.2%). The assessment of their antimicrobial activity against Candida albicans and six pathogenic bacteria revealed that O. libanoticum EO was inactive, while O. syriacum and O. ehrenbergii showed moderate antimicrobial activity with minimal inhibitory concentrations varying from 400 to 1200 μg/ml. These results support the traditional use of these last two species in traditional herbal preparations in Lebanon.     

Effect of volatile substances released fromOriganum majorana andOcimum basilicum on the rhizosphere and phyllosphere fungi ofPhaseolus vulgaris

Differences were found in the counts and occurrence of fungi in the phyllosphere and thizosphere of two representatives of the Lamiacea family,Origanum majorana andOcimum basilicum, and in the phyllosphere and rhizosphere ofPhaseolus vulgaris growing separately or in coenosis withO. majorana orO. basilicum. Both the volatile substances released from ground leaves of the two latter pantl species and the root exudates affected considerably spore germination of isolated phylospheric and rhizospheric fungi. The results indicated a possible role of root exudates and volatile substances released from leaves in colonization of rhizosphere and/or phyllosphere by fungi, especially in associations of various plants.     

The glycoflavone vicenin-2 its distribution in related genera within the labiatae

In a leaf flavonoid survey of Origanum the flavone C-glycoside, vicenin-2, was found to be restricted to the section Majorana. This is the first report of a glycoflavone in the Labiatae.     

Antifungal activity of essential oils against three vegetative-compatibility groups of <Emphasis Type="Italic">Verticillium dahliae</Emphasis>

The antifungal activities of volatile phase effects of essential oils from Origanum onites, O. syriacum, O. minutiflorum, O. vulgare, O, marjorana, Thymus vulgaris, T. serpyllum, Rosmarinus officinalis, Salvia officinalis and Micromeria fruticosa were evaluated for their ability to inhibit growth of three vegetative compatibility groups (VCGs) of Verticillium dahliae. Carvacrol was the main component of O. onites, O. minutiflorum and O. vulgare essential oils, while γ-terpinene was the main component of O. syriacum. P-cymene and thymol were the dominant component of T. vulgaris and T. serpyllum. β- thujone and l-camphor were the main component of S. officinalis. Polegone and isomenthone were the dominant components of M. fruticosa essential oil. Based on the in vitro test, the degree of fungistatical effects can be ranked in the following order of inhibition: O. syriacum = O. onites = O. minutiflorum = O. vulgare = T. vulgaris > T. serpyllum > M. fruticosa > S. officinalis = O. marjorana > R. officinalis. The essential oils of S. officinalis, O. marjorana and R. officinalis displayed moderate antifungal activity, that increased with increasing concentrations. Among the VCGs, VCG2A and VCG4B were found to be highly sensitive to the essential oils. The essential oils of O. syriacum, O. onites, O. minutiflorum, O. vulgare and T. vulgaris were the most efficacious, demonstrating strong antifungal activity against all of the tested VCGs of V. dahliae at relatively low concentrations and they could find practical application as natural fungicides in the prevention and protection of plants from V. dahliae infections.     

Plant odours with potential for a push–pull strategy to control the onion thrips, Thrips tabaci

We compared the efficacy of four plant essential oils to repel onion thrips, Thrips tabaci (Lindeman) (Thysanoptera: Thripidae), in the presence of an attractive odour, ethyl iso‐nicotinate in a pasture field. Four horizontal white sticky plates were placed adjacent to (directions: N, S, E, W) a central horizontal white plate (C). After 24 h, in the treatment combination where the four plates were sprayed with essential oil surrounding a central sticky plate sprayed with ethyl iso‐nicotinate, fewer onion thrips were found on the plates treated with sweet marjoram [Origanum majorana L. (Labiatae)] or clove basil [Ocimum gratissimum L. (Labiatae)] (87 and 71% less, respectively) compared to the control treatment of four water‐sprayed plates surrounding a central plate with ethyl iso‐nicotinate. We also compared the distribution of onion thrips on the plates. Relative thrips numbers on each plate were compared with similar (N, S, E, W, and C) plates in the control treatment. There were relatively lower thrips numbers on the south (23% reduction) and west (26% reduction) O. majorana‐treated plates and higher numbers (37% increase) on the central attractant‐treated plate indicating a short‐distance push–pull effect. When four plates were sprayed with the thrips attractant surrounding a central sticky plate sprayed with an essential oil or water (control), only O. majorana reduced the number of thrips on the attractant‐sprayed plates (62% reduction). The distribution of thrips on the different plates within this treatment combination did not change substantially when compared to the distribution in the water‐control treatment. Other essential oils tested (wormwood [Artemisia arborescens L. (Compositae)]) and tea tree [Melaleuca alternifolia (Maiden. & Betche.) Cheel. (Myrtaceae)]) were not effective in any of the treatments. It appears that O. majorana is a promising thrips repellent which could be used for further testing in a push–pull system with the attractant ethyl iso‐nicotinate. The field setup used also proved to be a valuable tool for evaluating the potential of repellents to control onion thrips.     

Bioactive properties of hydroalcoholic extract from Origanum onites L. as affected by glycerol incorporation

In the current study, the effect of glycerol -as a green solvent- addition into solvent mixture (50:50 ethanol:distilled water), on some biofunctional properties of Origanum onites L. was investigated. Response surface methodology (RSM) was used to detect the optimum conditions for the extraction process. Three variables namely glycerol concentration (X₁: 1–9 g), extraction temperature (X₂: 25–75 °C) and time (X₃:10–30 min) were selected and also total phenolic content, total flavonoid content, antioxidant capacity and antiradical activity of O. onites extracts were determined. Analysis of variance (ANOVA) showed that glycerol incorporation significantly increased the total phenolic content and antioxidant activity (p < 0.05) of the samples. Maximum levels to obtain the highest bioactive properties (highest total phenolic content and antioxidant activity) were determined as to be 9 g of glycerol addition for the extraction conditions as 45.4 °C and 75 min. This study reports the effect of glycerol on bioactive properties of O. onites and suggests that glycerol can be used to produce hydroalcoholic extracts having higher bioactivity from Origanum genus.     

Chemical Diversity of Wild Growing Origanum majorana in Cyprus

The present work aims at tracing the essential‐oil diversity of wild growing Origanum majorana plants in Cyprus. The study of six populations scattered over the western part of the island has shown that the essential oils were rich either in trans‐sabinene hydrate/terpinen‐4‐ol or in α‐terpineol/trans‐sabinene hydrate. The former oil type is more common and responsible for the typical marjoram odor of the plants. The comparison of our results with published data concerning wild and cultivated O. majorana showed that the oil composition of the Cypriot populations had not been reported from wild plants from other parts of the Mediterranean region, while it is very common among the cultivated plants. Thus, it seems probable that the nowadays cultivated marjoram may originate from Cypriot wild populations.     

Chemical composition and antifeedant activity of some aromatic plants against pine weevil (Hylobius abietis)

The pine weevil Hylobius abietis is an important pest causing severe damage to conifer seedlings in reforestation areas in Europe and Asia. Plants that have no evolutionary history with the pine weevil are of special interest in the search for compounds with a strong antifeedant activity. Thus, the essential oils of nine aromatic plants, viz Amomum subulatum, Cinnamomum tamala, Curcuma longa, Laurus nobilis, Ocimum basilicum, Origanum majorana, Origanum vulgare, Syzygium aromaticum and Trachyspermum ammi were extracted by hydrodistillation. The essential oil constituents were identified by gas chromatography–mass spectrometry, and antifeedant properties towards the pine weevil were assessed using choice feeding bioassay. The essential oils of C. longa, O. majorana, S. aromaticum and T. ammi showed an excellent antifeedant activity towards the pine weevil for 24 hr, whereas the essential oil of other plants showed the activity for 6 hr. There was a positive correlation between the amount of benzenoid compounds and the antifeedant activity of the essential oils. This study suggests that pine weevil non-host plant compounds have potential to be used for the protection of seedlings against pine weevil feeding. However, further study will be needed to explore the antifeedant activity of individual components and oils in the laboratory as well as in the field.     

Comparison of essential oils and genetic relationship of Origanum × intercedens to its parental taxa in the island of Crete

Plants of the taxa Origanum onites and Origanum vulgare were allowed to cross-pollinate under natural open field conditions and to produce a mixed population of putative hybrid Origanum × intercedens and parent genotypes. Randomly collected plants were classified as putative hybrids or parent genotypes by inspection of their inflorescence. They were then subjected to analysis of their essential oil composition and were fingerprinted by randomly amplified polymorphic DNA (RAPD) fragments. DNA primers identifying the genotype as well as showing the distance of a particular putative hybrid plant from the parent genotypes were found. Alone or in combination with the essential oil composition they can be used as reliable tools for the genetic identification of the two parental taxa and the putative hybrid plants in natural populations.     

Development of microsatellite markers for Diaphanosoma dubium (Crustacea,Cladocera) and application to seasonal population dynamics

Diaphanosoma, the “tropical Daphnia”, is common and ubiquitous in South China. Like other ctenopods, Diaphanosoma has a reproductive mode similar to Daphnia’s, but its resting eggs are rarely observed and lack an ephippium. With limited dispersal and reduced buffer effect from resting egg banks, Diaphanosoma is expected to have a population genetic structure different from that of temperate Daphnia. To facilitate genetic comparison, we developed microsatellite markers using next-generation sequencing for the most common species in tropical and subtropical East Asia, Diaphanosoma dubium. Thirty-one polymorphic microsatellite markers were obtained, and 29 of them were efficient for the congeneric species D. excisum, D. orghidani, D. mongolianum and D. chankensis. The markers allowed intra- and interspecific genetic analysis, including population structure, hybridization and introgression. We used 11 selected microsatellite markers to analyze spatial and temporal heterogeneity of genetic diversity in four (sub)tropical D. dubium populations from two large reservoirs and two temporary ponds. In contrast to temperate Cladocera, higher genetic diversity in summer rather than in spring suggested weak contribution from resting eggs in spring. Clustering of DAPC and STRUCTURE analyses indicated a clear-cut genetic structure in the four populations. Variation partitioning revealed that water storage and depth were key factors in genetic differentiation. Within large reservoirs, we detected backward (reversing time) gene flow from resting egg banks. We conclude that resting eggs have an effective contribution to the genetic diversity in large water bodies during growing seasons and that large water bodies can host higher genetic diversity in summer due to environmental heterogeneity and high carrying capacity. Spatial and temporal heterogeneity of genetic diversity detected by our microsatellite markers showed the newly developed markers can be applied for further study of populations of D. dubium and other species of Diaphanosoma at a contemporary scale.

     

Effect of volatile substances fromOriganum majorana andOcimum basilicum on spore respiration and germination of some soil fungi

Oxygen uptake by the spores ofFusarium moniliforme, F. oxysporum, F. semitectum, F. solani, Mucor racemosus andTrichoderma viride was increased in the presence of volatile substances extracted, fromOriganum majorana andOcimum basilicum. This increase was greater in the presence of volatile substances fromO. basilicum thanO. majorana, except in the case ofF. semitectum where the reverse was true. A drop in the RQ of all the germinating spores was observed in the presence of these substances. Volatile substances fromO. majorana reduced the spore germination ofM. racemosus whereas the spores ofT. viride were stimulated to germinate. Volatile substances fromO. basilicum stimulated the spore germination ofM. racemosus whereasT. viride spores were not affected.     

Medicinal Uses,Phytochemistry, and Pharmacology of Origanum onites (L.): A Review

Origanum onites L., known as Turkish oregano, has great traditional, medicinal, preservative, and commercial importance. It is used for the treatment of several kinds of ailments, such as gastrointestinal disorders, diabetes, high cholesterol, leukemia, bronchitis, etc. In this review, traditional use, phytochemistry, and pharmacology of O. onites reported between 1988 and 2014 were discussed. This review was prepared based on literature survey on scientific journals and books from libraries and electronic sources, such as Web of Science, PubMed, Scopus, Google Scholar, etc. All databases were searched up to June 2014. Several different classes of terpenoids, triterpene acids, phenolic acids, hydroquinones, flavonoids, hydrocarbons, sterols, pigments, fatty acids, tocopherols, and inorganic compounds were detected mainly in the aerial parts of this plant. Pharmacological studies revealed that extracts obtained from several solvents and individual compounds exhibited antimicrobial, antiviral, antioxidant, insecticidal, anticancer, hepatoprotective, genotoxic, antidiabetic, cholinesterase inhibitory, anti‐inflammatory, analgesic activities, etc. O. onites, in general, exhibited remarkable activity potential in almost all test systems. The results of toxicity studies indicated that O. onites did not show any significant toxicity and mutagenicity on Drosophila and Salmonella. Toxicity of the extracts/essential oils and also individual compounds should be evaluated on mammalian cells to ensure their safety. The bioactivity of individual compounds aside from terpenoids should also be assessed in detail. Additionally, mode of action for the bioactive compounds should be evaluated to understand the complex pharmacological effects of these phytochemicals.     

New chromosome counts in the genus Centaurea (Asteraceae) from Turkey

Twenty‐two chromosome counts are reported in 16 species, four subspecies and two varieties of the genus Centaurea. These are mostly Turkish local endemics of section Cheirolepis, a complicated group from the Eastern clade of the Jacea group. Twenty‐one reports are new. Prevalence of the basic chromosome number x = 9 among the eastern sections of the Jacea group is confirmed. © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 159 , 280–286.     

Environmental (in)dependence of a hybrid zone: Insights from molecular markers and ecological niche modeling in a hybrid zone of Origanum (Lamiaceae) on the island of Crete

The role of environment and the relative significance of endogenous versus exogenous selection in shaping hybrid zones have been crucial issues in the studies of hybridization. Recent advances in ecological niche modeling (ENM) offer new methodological tools, especially in combination with the genotyping of individuals in the hybrid zone. Here, we study the hybrid zone between the widely known spices Origanum onites and Origanum vulgare ssp. hirtum in Crete. We analyze the genetic structure of both parental taxa and their hybrid Origanum × intercendens using AFLP markers on 15 sympatric and 12 allopatric populations and employ ecological niche modeling and niche similarity tests to study their niche patterns. We complement these analyses with seed viability measurements. Our study revealed that the hybridizing taxa O. onites and O. vulgare ssp. hirtum and the resulting genotypic classes showed geographical and environmental niche similarities based on the predictions of ENMs and the subsequent similarity tests. The occurrence of the hybrid zone is not directly dependent on environmental factors which favor the fitness of the hybrid compared to the parental taxa, but rather on aspects such as historical factors and management practices, which may contribute to the localization and maintenance of the contact zone between parental species. Our results suggest that if a minimum required niche differentiation between genotypic classes is not achieved, environmental dependence might not have a prominent role on the outcome of the hybridization.     

Species delimitation in Lippia section Goniostachyum (Verbenaceae) using the phylogenetic species concept

Lippia section Goniostachyum comprises plants distinguished by their numerous axillary florescences (three to six, sometimes up to nine) and tetrastichous floral bracts. Species of section Goniostachyum occur in the Neotropics, from Mexico to northern Argentina. Delimitation of the species grouped under Goniostachyum has remained unclear. Forty‐one names exist under this section, but only c. eight to ten names have been used frequently. To resolve the taxonomy of this group, a modified population aggregation analysis, based on the phylogenetic species concept, was employed. As a result, Goniostachyum is here circumscribed to only four species: L. grata, L. origanoides, L. sericea and L. stachyoides. These species are supported by different combinations of three characters of the 13 qualitative attributes analysed: canescent sericeous pubescence, frondose or frondose‐bracteose inflorescences and free or fused florescence apical bracts. Two varieties based on significant differences among quantitative characters are recognized: L. stachyoides var. stachyoides and L. stachyoides var. martiana comb. nov. Fifteen lectotypifications and four neotypifications are proposed. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 170 , 197–219.     

A taxonomic review of the catfish identified as Glyptothorax zanaensis (Teleostei: Siluriformes: Sisoridae), with the descriptions of two new species

This study re‐examined the taxonomic status of the sisorid catfishes usually identified as Glyptothorax zanaensis using a combination of morphometric and molecular data. Our results resurrect Glyptothorax longinema from the synonymy of G. zanaensis, and we describe two previously unnamed species as Glyptothorax granosus sp. nov. and Glyptothorax fucatus sp. nov. All four species are diagnosed and described in detail. Truss‐based morphometrics combined with principal component analysis (PCA) detected three principal components (PCs) that can explain 86% of the total variation amongst species, which mainly reflect the characteristics of body depth, related depth, adhesive apparatus length, pectoral‐fin length, caudal peduncle length, and barbel lengths. We also generated a phylogenetic hypothesis of these species using concatenated mitochondrial cytochrome b and d ‐loop gene sequences. Molecular dating analysis revealed a rapid speciation of Glyptothorax in the south‐eastern corner of the Qinghai‐Xizang Plateau from the middle Pliocene to early Pleistocene. A key to identify the Glyptothorax species from the Salween River drainage is also provided. © 2012 The Linnean Society of London, Zoological Journal of the Linnean Society, 2012, 165 , 363–389.     

A comparative study of cytotoxic, membrane and DNA damaging effects of Origanum majorana’s essential oil and its oxygenated monoterpene component linalool on parental and epirubicin-resistant H1299 cells

In this study, cytotoxic, membrane and DNA damaging effects of the essential oil from Origanum majorana and its oxygenated monoterpene component linalool were tested on parental and epirubicin-resistant (drug-resistant) human lung cancer cell lines (H1299). Essential oil’s and linalool’s cytotoxicities were examined and parental cells were found more sensitive to the essential oil’s and linalool’s cytotoxicities than drug-resistant cells. O. majorana essential oil had more effective membrane damaging effect than linalool on parental cells, while in drug-resistant H1299 cells, linalool had more effective membrane damaging effect than the essential oil. O. majorana essential oil possessed more effective DNA damaging effect than linalool on both parental and drug-resistant cells. The conclusions from this study suggest that O. majorana essential oil and linalool exhibit cytotoxic, membrane and DNA damaging effects. They thus need further investigation as potential therapeutic agents for human lung cancer.     

Plant growth promoting rhizobacteria improve growth and essential oil yield in Origanum majorana L.

Effects of root colonization by plant growth promoting rhizobacteria (PGPR) on biomass, and qualitative and quantitative composition of essential oils, were determined in the aromatic crop Origanum majorana L. (sweet marjoram). PGPR strains evaluated were Pseudomonas fluorescens, Bacillus subtilis, Sinorhizobium meliloti, and Bradyrhizobium sp. Only P. fluorescens and Bradyrhizobium sp. showed significant increases in shoot length, shoot weight, number of leaf, number of node, and root dry weight, in comparison to control plants or plants treated with other PGPR. Essential oil yield was also significantly increased relative to non-inoculated plants, without alteration of oil composition. P. fluorescens has clear commercial potential for economic cultivation of O. majorana.     

Nutlet micromorphology of Carex section Rhomboidales sensu Kükenthal (Cyperaceae) and its systematic implications

The sectional delimitation of Carex section Rhomboidales sensu Kükenthal (1909) is problematic and, here, we provide contributions to a systematic understanding of nutlet micromorphology in section Rhomboidales. Nutlet micromorphology of 87 samples representing 71 taxa was investigated using scanning electron microscopy and nine nutlet types were recognized. Although some closely related taxa show similarities, nutlet morphology in section Rhomboidales is stable at the species level and is reliable for species identification. Based on the nutlet characters, section Rhomboidales is delimited and emended as having nutlet rhombic–ovoid, trigonous; beaks conspicuous, erect, rarely curved or coiled; periclinal walls of epidermal cells straight, with zero to two silica bodies. The species with nutlets of the C. harlandii‐type and C. thibetica‐type are included, and species of the C. chinensis‐type and C. macrandrolepsis‐type are ascribed to section Mitratae and section Infossae, respectively. Carex longirostrata and C. pseudolongirostrata (= C. nodaeana), formerly ascribed to section Careyanae or section Depauperatae, are well supported as members of section Rhomboidales. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 175 , 123–143.