Antifungal properties of ethanolic extract and its active compounds from Calocedrus macrolepis var. formosana (Florin) heartwood

The ethanolic extract of Calocedrus macrolepis var. formosana heartwood was screened for antifungal compounds by agar dilution assay and liquid chromatography. Two compounds, beta-thujaplicin and gamma-thujaplicin, responsible for the antifungal property of C. macrolepis var. formosana heartwood were isolated by high performance liquid chromatography (HPLC), and identified by 1H NMR and 13C NMR. The antifungal activities of these two compounds were further evaluated against total 15 fungi, including wood decay fungi, tree pathogenic fungi and molds. The hexane soluble fraction showed the strongest antifungal activities among all fractions. beta-Thujaplicin and gamma-thujaplicin exhibited not only very strong antifungal activity, but also broad antifungal spectrum. The MIC values of beta-thujaplicin and gamma-thujaplicin were in the range of 5.0-50.0 microg/ml. In addition, scanning electron microscopy (SEM) was carried out to study the structural change of fungal hyphae induced by beta-thujaplicin. Strong cell wall shrinkage indicated the fungicidal effect could be attributed to the combined actions of metal chelating and cytoplasm leakage. It also suggests that the role of metal chelating is indispensable in the design of environmental-friendly fungicides.     

Phylogenetic relationships of the genus Chamaecyparis inferred from leaf essential oil

The species differentiation between Chamaecyparis formosensis, C. obtusa var. formosana, and C. obtusa, based on the composition of the leaf essential oils, was studied. The characterization of the oils by GC-FID and GC/MS analyses showed remarkable differences between these three essential oils. Cluster analysis (CA) and principal-component analysis (PCA) distinguished three groups of essential oils. The C. formosensis oil was dominated by α-pinene while those isolated from C. obtusa var. formosana and C. obtusa were characterized by high levels of (-)-thujopsene and α-terpinyl acetate, respectively. Moreover, the phylogenetic relationships of the genus Chamaecyparis were in agreement with previous findings based on morphological and molecular evidence. In addition, the essential oils from C. obtusa var. formosana could be classified into three chemical types, according to their different characteristic main compounds (β-elemol, (-)-thujopsene, and cis-thujopsenal). The biochemical correlations between the major constituents of the Chamaecyparis species were examined and their relationship is discussed.     

Antifungal activity of essential oil and its constituents from Calocedrus macrolepis var. formosana Florin leaf against plant pathogenic fungi

Resistance to conventional fungicides causes the poor disease control of agriculture. Natural products from plants have great potential as novel fungicide sources for controlling pathogenic fungi. In this study antipathogenic activity of the leaf essential oil and its constituents from Calocedrus macrolepis var. formosana Florin were evaluated in vitro against six plant pathogenic fungi. Chemical analysis of leaf oil by GC/MS allowed identification of alpha-pinene (44.2%), limonene (21.6%), beta-myrcene (8.9%), beta-caryophyllene (8.2%), caryophyllene oxide (2.4%), alpha-cadinol (1.6%), beta-pinene (1.2%), and T-muurolol (1.1%) as main components. Sesquiterpenoid components of the oil were more effective than monoterpenoid components of the oil. In particular, T-muurolol and alpha-cadinol strongly inhibited the growth of Rhizoctonia solani and Fusarium oxysporum, with the IC(50) values < 50 microg ml(-1). These compounds also efficiently inhibited the mycelial growths of Colletotrichum gloeosporioides, P. funerea, Ganoderma australe and F. solani. These results showed that T-muurolol and alpha-cadinol possess antifungal activities against a broad spectrum of tested plant pathogenic fungi and could be used as potential antifungal agents for the control of fungal diseases in plants.     

北细辛挥发油对二斑叶螨的毒性及其酶的影响

采用气相/质谱分析了北细辛挥发油的化学成分,并研究了北细辛挥发油对二斑叶螨的薰蒸毒性,以及对谷胱甘肽巯基转移酶(GSTs)与Na+,K+-ATP酶活性的影响。结果表明:有9种含量大于1%的化学成分占北细辛挥发油总量的86.1%,其中甲基丁香酚含量最高(22.9%),其次为黄樟醚(19.8%);北细辛挥发油在8μg/mL时对二斑叶螨的熏蒸毒性最强,其24和48 h的螨死亡率分别为72.6%和100%;以4μg/mL北细辛挥发油薰蒸,在24 h内螨Na+,K+-ATP酶活性下降、GSTs活性升高。本研究结果提示北细辛挥发油有开发为农业用杀螨剂的潜在价值。     

小花假泽兰精油超临界CO_2提取工艺及其成分和抑菌活性研究

采用正交试验法研究超临界CO_2萃取法提取小花假泽兰精油的提取条件,采用GC-MS法分析测定精油的化学成分,并初步测定精油的抑菌效果.结果表明:超临界CO_2萃取小花假泽兰精油的较佳工艺条件为:采用先静态后动态的萃取方式,静态萃取时间20 min,动态萃取条件为萃取温度55℃,萃取压力35 Mpa,CO_2体积为40 mL/g,CO_2 流速为0.5～1.0 mL/min;从超临界CO_2萃取的小花假泽兰精油中鉴定了64种成分,占精油总量的79.80%,主要有萜类、醇类、脂肪酸、酯类、甾体类等;生物测定结果表明,超临界CO_2萃取物对小麦赤霉病菌和小麦纹枯病菌菌丝生长的EC_(50)分别为119.55 mg/L和78.27 mg/L. 果.结果表明:超临界CO_2萃取小花假泽兰精油的较佳工艺条件为:采用先静态后动态的萃取方式,静态萃取时间20 min,动态萃取条件为萃取温度55℃,萃取压力35 MPa,CO_2体积为40 mL/g,CO_2 流速为0.5～1.0 mL/min;从超临界CO2萃取的小花假泽兰精油中鉴定了64种成分,占精油总量的79.80%,主要有萜类、醇类、脂肪酸、酯类、甾体类等;生物测定结果表 ,超临界CO_2萃取物对     

Antimicrobial activity of a traditionally used complex essential oil distillate (Olbas(?) Tropfen) in comparison to its individual essential oil ingredients

Plant extracts and essential oils have been widely studied and used as antimicrobial agents in the last decades. In our study we investigated the antimicrobial activities of Olbas(?) Tropfen (in the following named Olbas), a traditionally used complex essential oil distillate, in comparison to its individual essential oil ingredients. Olbas (10g) consists of three major components such as peppermint oil (5.3g), eucalyptus oil (2.1g), and cajuput oil (2.1g) and of two minor constituents like juniper berry oil (0.3g) and wintergreen oil (0.2g). The composition of Olbas and the five individual essential oils were characterized by GLC-MS. According to GLC-MS analysis 1,8-cineol is the main component of the complex essential oil distillate followed by menthol and menthone. The minimum inhibitory and minimum microbicidal concentrations of Olbas and each of the single essential oils were evaluated in 17 species/strains of bacteria and fungi. Time-kill assay was performed to compare the microbicidal activity of Olbas and peppermint oil during several time intervals. Olbas displayed a high antimicrobial activity against all test strains used in this study, among them antibiotic resistant MRSA (methicillin-resistant Staphylococcus aureus) and VRE (vancomycin-resistant Enterococcus). Its antimicrobial activity was comparable to that of peppermint oil which was the most potent one of all individual essential oils tested. In the time kill assay Olbas as well as peppermint oil demonstrated similar microbicidal activities. Based on its wide antimicrobial properties Olbas can be a useful agent for the treatment of uncomplicated infections of skin and respiratory tract.     

Antifungal Activity of the Essential Oils of Callitris neocaledonica and C. sulcata Heartwood (Cupressaceae)

Mortality due to fungal infections has increased substantially, becoming a worldwide problem in public health. As a contribution to the discovery of new antifungal agents, the properties of the heartwood essential oils of two trees growing in New Caledonia, Callitris neocaledonica and C. sulcata (Cupressaceae) were investigated. The essential oils extracted by hydrodistillation were characterized by GC-FID and GC/MS analyses. From C. neocaledonica oil, 31 constituents were identified, representing 97.0% of the total oil composition, which was mainly constituted by oxygenated sesquiterpenes (88%). Among them, guaiol (1; 30.2%), bulnesol (2; 12.5%), α-eudesmol (3; 10.5%), β-eudesmol (4; 10.5%), γ-eudesmol (10.2%), and elemol (4.9%) predominated. The chemical composition of C. sulcata oil, from which 39 constituents were identified (96.8% of the total oil composition), showed some similarities with that of C. neocaledonica oil. The major constituents were also oxygenated sesquiterpenes, accounting for 78.5% of the oil, amongst them, mainly compounds 1 (16.1%), 3 and 4 (9.7% each), as well as 2 (7.4%). The antifungal activity of the oils against clinical isolates of four dermatophytic fungi (Trichophyton mentagrophytes, T. rubrum, Microsporum canis, and M. gypseum) and six yeasts (Candida albicans, C. parapsilosis, C. glabrata, C. krusei, Cryptococcus neoformans, and Cryptococcus gattii) was tested by determining minimum inhibitory concentrations (MICs) using the microdilution method. The best antifungal activities of the C. neocaledonica and C. sulcata oils were obtained against C. krusei (MICs of 3.9 and 0.975?μg/ml, resp.). These MIC values were similar to those of the reference drugs itraconazole and fluconazole (1.0 and 0.5?mg/ml, resp.). The oils were also subjected to a screening for their possible DPPH(.) (2,2-diphenyl-1-picrylhydrazyl) radical-scavenging activity. C. neocaledonica essential oil was more active than C. sulcata oil (93.3 vs. 32.2% DPPH(.) scavenged at 250?μg/ml).     

Chemical composition and oxidative stability of flax, safflower and poppy seed and seed oils

Three seeds of Turkish origin, flax, poppy and safflower were analyzed for their proximate, fatty acids, tocols (tocopherols and tocotrienols) and total phenolic composition, and oxidative stability of their oil. The major fatty acid in the flax oil was alpha-linolenic acid, comprising 58.3% of total fatty acids, whereas poppy and safflower oils were rich in linoleic acid at 74.5% and 70.5% level, respectively. The amount of total tocols was 14.6 mg/100g flax, 11.0mg/100g poppy and 12.1mg/100g safflower seed. Flax and poppy oil were rich in gamma-tocopherol as 79.4 mg/100g oil and 30.9 mg/100g oil, respectively, while alpha-tocopherol (44.1g/100g oil) was dominant in safflower oil. Only alpha- and gamma-tocotrienol were found in the oils. Oxidative stability of oils was measured at 110 degrees C at the rate of 20 L/h air flow rate, and poppy oil (5.56 h) was most stabile oil followed by safflower oil (2.87 h) and flax oil (1.57). There were no correlation between oxidative stability and unsaturation degree of fatty acids and tocol levels of the oils. All of the seeds investigated provide a healthy oil profile and may have potential as a source of specialty oils on a commercial scale.     

Chemical composition and antitermitic activity against Coptotermes formosanus Shiraki of Cryptomeria japonica leaf essential oil

Cryptomeria japonica D. Don, called 'sugi' in Japanese, is an abundant and renewable potential resource of valuable natural products that may serve as natural biocides in Taiwan. Hydrodistillation (HD) and steam distillation (SD) were used for extracting the essential oils from C. japonica leaves in this study. The chemical constituents of the two leaf essential oils were identified by GC/MS analysis, and their antitermitic activities were evaluated. The results from the antitermitic tests against Coptotermes formosanus Shiraki showed that the leaf essential oils extracted by HD and SD had excellent antitermitic activities, with LD(50) values of 1.57 and 1.72 mg/g after 7 d of testing, respectively. Comparison of the chemical composition of the two leaf essential oils revealed that oxygenated diterpenes (kaur-16-ene) and oxygenated sesquiterpenes (β-elemol) were the dominant constituents. In addition, 13 constituents isolated from the C. japonica leaf essential oils were tested individually against C. formosanus. Among these constituents, β-elemol and α-terpineol achieved 100% termite mortality at the dosage of 1 mg/g after 7 d of testing. These results demonstrated that both C. japonica leaf essential oils as well as β-elemol and α-terpineol possessed commendable antitermitic activity.     

Antioxidant properties of essential oils: autoxidation of essential oils from laurel and fennel and effects of mixing with essential oil from coriander

Changes in the composition of essential oils from the seeds of laurel (Laurus nobilis L.) and fennel (Foeniculum vulgare Mill., var. dulce Thelling) and their mixture with essential oil from coriander were studied by capillary gas-liquid chromatography during storage in the dark and in light. Under these conditions, essential oil of laurel retained its composition for 12 months. Essential oil of fennel was rapidly oxidized in light. However, the rate of its oxidation in the dark was lower. The major component of essential oil of fennel, transanethol, had a lower antioxidant activity than essential oil of coriander. The mixture of essential oils from laurel and coriander possessed antioxidant properties and strongly inhibited the oxidation of components of the fennel oil.     

Chemical Composition of the Leaf and Flower Essential Oils of Tunisian Lavandula dentata L. (Lamiaceae)

Essential oils of Lavandula dentata, a Tunisian native plant, were isolated from leaves and flowers by hydrodistillation in a Clevenger‐type apparatus and characterized by GC‐FID and GC/MS analyses. The average essential oil yields, means of five replicates, were higher for the flowers (8.60 mg/g) than for the leaves (6.56 mg/g). A total of 72 compounds were identified, accounting for 98.1 and 97.7% of the total oil composition of the leaves and flowers, respectively. The main essential oil constituents were 1,8‐cineole, camphor, and L ‐fenchone, accounting for 33.54, 18.89, and 8.36% in the leaf oils and for 19.85, 23.33, and 7.13% in the flower oils, respectively. Besides this quantitative variation, the results also showed considerable qualitative variation between the essential oils of the two plant parts analyzed. These differences might be adaptative responses to ecological exigencies.     

Effects of Cymbopogon nardus (L.) W. Watson essential oil on the growth and morphogenesis of Aspergillus niger

The growth inhibitory effect of Cymbopogon nardus (L.) W. Watson var. nurdus essential oil on Aspergillus niger (Van Tieghem) mycelium was determined on agar medium. The mycelium growth was completely inhibited at 800 mg/L. This concentration was found to be lethal under the test conditions. Essential oil at 400 mg/L caused growth inhibition of 80% after 4 days of incubation, and a delay in conidiation of 4 days compared with the control. Microscopic observations were carried out to determine the ultrastructural modifications of A. niger hyphae after treatment with C. nardus essential oil. The main change observed by transmission electron microscopy concerned the hyphal diameter and the hyphal wall, which appeared markedly thinner. These modifications in cytological structure might be caused by the interference of the essential oil with the enzymes responsible for wall synthesis which disturb normal growth. Moreover, the essential oil caused plasma membrane disruption and mitochondrial structure disorganization. The findings thus indicate the possibility of exploiting Cymbopogon nardus essential oil as an effective inhibitor of biodegrading and storage-contaminating fungi.     

Bioactivity assay of extracts from Calocedrus macrolepis var. formosana bark

Alcoholic extracts from bark of Calocedrus macrolepis var. formosana Florin (Cupressaceae) were extracted successively using n-hexane, dichloromethane, ethyl acetate, 1-butanol and water, which gave 34.8%, 34.1%, 24.1%, 3.3% and 3.7% soluble fractions, respectively. Antioxidation activity of these fractions by DPPH assay and dissimilar IC50 values of the DPPH showed that ethyl acetate fraction had the best antioxidant activity; its IC50 was 2.6 microg/ml. Analyses of the composition and anti-inflammatory activity of the subfractions from n-C6H14 fraction showed that the T3 and H5ppt had the best anti-inflammatory activity in LPS-stimulated murine macrophage J774A. 1 cells, respectively; moreover, their major constituent was sugiol (T3 37.1%, H5ppt 81.1%), which at dosages of 10 microg/ml inhibited proIL-1beta protein production completely. Furthermore, the T1 also exhibited anti-inflammatory activity, and its major constituent was ferruginol (above 85.6%).     

Vapor phase toxicity of plant essential oils to Cadra cautella (Lepidoptera: Pyralidae)

The toxicities of 44 plant essential oils against larvae of Cadra cautella (Walker) were examined using direct contact and vapor phase toxicity bioassays and compared with the lethal activity of chlorpyrifos-methyl, diazinon, dichlorvos, and fenthion, four widely used organophosphorus insecticides. Responses varied according to plant material used and exposure dose and time. In a filter paper contact toxicity bioassay, potent toxicity was produced from buchu leaf, niaouli, and rosemary oils at 2.4 mg/cm2 and armoise, cypress, galbanum, and mace oils at 4.7 mg/cm2. In vapor phase toxicity bioassays with larvae, cypress, galbanum, niaouli, and rosemary oils were much more effective in closed containers than in open containers, indicating that the lethal effects of these oils were largely because of action in the vapor phase. As judged by 24-h LC50 values, potent fumigant action was observed with niaouli oil (64.7 mg/liter air) and rosemary oil (64.6 mg liter/air). Cypress and galbanum oils exhibited weak fumigant activity. These essential oils were less active than dichlorvos (0.86 mg/liter air). Little or no fumigant action was observed with chlorpyrifos-methyl, diazinon, and fenthion. Essential oils described herein, particularly niaouli and rosemary oils, merit further study as potential larvicides for the control of C. cautella.     

Composition and the in vitro antimicrobial activities of the essential oils of some Thymus species

The genus Thymus (Lamiaceae) is represented by 38 species (64 taxa) in Turkey, and 24 of which are endemic to Turkey. Aerial parts of Thymus longicaulis subsp. chaubardii var. chaubardii, T. zygioides var. lycaonicus, T. longicaulis subsp. longicaulis var. subisophyllus and T. pulvinatus collected from three different localities in Balikesir province were subjected to hydrodistillation to yield essential oils which were subsequently analysed by GC and GC/MS. The main constituents of the oils were identified, and antimicrobial bioassay was applied. Thymol (56.6%, 42.8%, 36.9%) was the main component in the oils of T. longicaulis subsp. chaubardii var. chaubardii (chemotype I), T. longicaulis subsp. chaubardii var. chaubardii (chemotype II) and T. zygioides var. lycaonicus respectively. The oil of T. longicaulis subsp. longicaulis var. subisophyllus contained carvacrol (60.0%) and the oil of T. pulvinatus borneol (27.9%) as main constituents.     

华南五种木兰科植物精油成分和抗氧化活性

对广东南岭自然保护区内野生的毛桃木莲（Manglietia moto）、乳源木莲（M．yuyanensis）、乐昌含笑（M．chapensis）、金叶含笑（M．foveolata）和深山含笑（M．maudiae）5种木兰科芳香植物精油进行了化学成分对比分析和抗氧化活性研究。化学成分鉴定采用气质联用技术,同时结合了保留指数比较法;精油抗氧化活性采用β-胡萝卜素漂白试验法。深山含笑芳香精油的主体成分以单萜烯为主,另外4种木兰科芳香精油所含成分均以倍半萜类为主。毛桃木莲和乳源木莲精油在成分构成上具有高度一致性,而且在主要成分上具有相似性,这可能表明它们具有非常近的植物亲缘关系。4种精油的抗脂质氧化半抑制浓度分别为6．6g／L（毛桃木莲精油）、9．8g／L（乳源木莲精油）、11．3g／L）金叶含笑精油）和12．2g／L（乐昌含笑精油）,在本试验条件下,未能测出深山含笑的半抑制浓度。     

Antioxidant Properties of Essential Oils: Autoxidation of Essential Oils from Laurel and Fennel and of Their Mixtures with Essential Oil from Coriander

Changes in the composition of essential oils from the seeds of laurel (Laurus nobilis L.) and fennel (Foeniculum vulgare Mill., var. dulce Thelling) and their mixture with essential oil from coriander were studied by capillary gas-liquid chromatography during storage in the dark and in light. Under these conditions, essential oil of laurel retained its composition for 12 months. Essential oil of fennel was rapidly oxidized in light. However, the rate of its oxidation in the dark was lower. The major component of essential oil of fennel, trans-anethol, had a lower antioxidant activity than essential oil of coriander. The mixture of essential oils from laurel and coriander possessed antioxidant properties and strongly inhibited the oxidation of components of the fennel oil.     

杉木心材精油抑菌活性及其化学成分研究

通过水蒸气蒸馏法提取杉木心材精油,并进行柱层析分离、气-质联用分析和抑菌活性试验,比较分析了精油含量、化学组成和抑菌活性成分.结果表明,杉木心材精油含量为1.794～2.076(w/w);气-质联用分析共分离出47个色谱峰,鉴定出27个化合物(占精油总量的99%),其中主要成分为柏木脑(76.27%);杉木心材精油对大肠杆菌、金黄色葡萄球菌、枯草芽孢杆菌、伤寒沙门氏菌等均有较明显的抑制作用;柏木脑是杉木精油的主要抑菌活性成分.     

The synergism of antifungals and essential oils against Candida spp. evaluated by a modified gradient-diffusion method

The usefulness of modified method of MIC Test Strip, for determining the synergistic effect of essential oils in the liquid or volatile phase with fluconazole and voriconazole, was evaluated. Geranium oil used against C. albicans in agar dilution test, at a concentration of 1/2 MIC caused a drop in the value of fluconazole MIC from 12.0 mg/l to 0.064 mg/l and voriconazole from 0.125 mg/l to 0.006 mg/l. A similar effect of drug combinations with essential oils was obtained in the case of C. glabrata study. Volatile Clove oil and cytronelal, applied in subMIC concentrations, also caused a reduction offluconazole and voriconazole MICs. Thus, utility of this simple methods developed by us for testing the effectiveness of combinations of known drugs and new compounds with antifungal activity, has been confirmed.     

Repellent and acaricidal activities of basil (Ocimum basilicum) essential oils and rock dust against Ixodes scapularis and Dermacentor variabilis ticks

Repellent and acaricidal activity of essential oils extracted from three varieties of basil (Ocimum basilicum L.) were evaluated on blacklegged ticks (Ixodes scapularis Say) and American dog ticks (Dermacentor variabilis Say) in laboratory conditions. Essential oils were extracted and characterized through gas chromatography-mass spectrometry, and tested at different concentrations for long-term repellent activity using horizontal bioassays. In addition, basil essential oils were combined with an inert material (i.e., granite rock dust) with known insecticidal and miticidal properties to assess acaricidal activities against adult ticks. Among the tested basil varieties, var. Jolina essential oil at 15% vol/vol concentration repelled 96% of tested ticks up to 2 h post-treatment. The EC₅₀ for I. scapularis nymphs was 4.65% vol/vol (95% confidence interval: 4.73–4.57). In acaricidal tests, the combination of essential oil from var. Aroma 2 at 10% wt/wt with rock dust resulted in 100% tick mortality after only 24 h post-exposure, with a LD₅₀ of 3.48% wt/wt (95% CI 4.05–2.91) for freshly prepared treatment tested on I. scapularis adults. The most common compounds detected in basil essential oils by GC–MS were linalool (52.2% in var. Nu Far, 48.2% in Aroma 2, 43.9% in Jolina), sabinene (6.71% in Nu Far, 8.99% in Aroma 2, 8.11% in Jolina), eugenol (11.2% in Jolina, 8.71% in Aroma 2), and estragole (18.2% in Nu Far). The use of essential oils alone and in combination with rock dust provides an innovative and environmentally friendly approach for managing ticks and inhibiting vector-borne disease transmission.