Composition and biological activities of essential oils from Limnophila geoffrayi Bonati

Essential oils from the dried aerial parts of Limnophila geoffrayi Bonati were obtained by water-distillation. d-Pulegone (27.14%), perillaldehyde (19.13%) and limonene (9.00%) were characterized as the major constituents using gas chromatography-mass spectrometry analysis. The antimicrobial activities of the essential oils and their major components were evaluated against microorganisms encountered normally in contaminated cosmetic products, using the agar- and broth-dilution methods. Their insecticidal activities against the Oriental fruit fly Bactrocera dorsalis (Hendel) were tested using a bioassay with impregnated filter paper. The results showed that the essential oils possessed high antimicrobial activity, with minimum inhibitory concentrations ranging from 0.03 to 0.2% per unit volume (v/v). Strong insecticidal activity as a fumigant was also observed at an oil dose of 5 μl/disc, with a 94% mortality. Perillaldehyde was the most active compound among the main components of these essential oils.     

Antimicrobial Activities of the Essential Oils of Various Plants against Tomato Late Blight Disease Agent Phytophthora infestans

The aim of this study was to find an alternative to synthetic fungicides currently used in the control of devastating oomycete pathogen Phytophthora infestans, causal agent of late blight disease of tomato. Antifungal activities of essential oils obtained from aerial parts of aromatic plants such as oregano (Origanum syriacum var. bevanii), thyme (Thymbra spicata subsp. spicata), lavender (Lavandula stoechas subsp. stoechas), rosemary (Rosmarinus officinalis), fennel (Foeniculum vulgare), and laurel (Laurus nobilis), were investigated against P. infestans. Both contact and volatile phase effects of different concentrations of the essential oils used were determined by using two in vitro methods. Chemical compositions of the essential oils were also determined by GC-MS analysis. Major compounds found in essential oils of thyme, oregano, rosemary, lavender, fennel and laurel were carvacrol (37.9%), carvacrol (79.8), borneol (20.4%), camphor (20.2%), anethole (82.8%) and 1,8-cineole (35.5%), respectively. All essential oils were found to inhibit the growth of P. infestans in a dose-dependent manner. Volatile phase effect of oregano and thyme oils at 0.3 μg/ml air was found to completely inhibit the growth of P. infestans. Complete growth inhibition of pathogen by essential oil of fennel, rosemary, lavender and laurel was, however, observed at 0.4–2.0 μg/ml air concentrations. For the determination of the contact phase effects of the tested essential oils, oregano, thyme and fennel oils at 6.4 μg/ml were found to inhibit the growth of P. infestans completely. Essential oils of rosemary, lavender and laurel were inhibitory at relatively higher concentrations (12.8, 25.6, 51.2 μg/ml respectively). Volatile phase effects of essential oils were consistently found to be more effective on fungal growth than contact phase effect. Sporangial production was also inhibited by the essential oil tested. Light and scanning electron microscopic (SEM) observation on pathogen hyphae, exposed to both volatile and contact phase of oil, revealed considerable morphological alterations in hyphae such as cytoplasmic coagulation, vacuolations, hyphal shrivelling and protoplast leakage.     

Antibacterial activities of essential oils from eight Greek aromatic plants against clinical isolates of Staphylococcus aureus

Aromatic plants have been used widely to extend the shelf life of foods but at the same time research is undergoes for their properties as antibacterial agents in clinical use. Although there are promising results for the antimicrobial properties of various essential oils against environmental or food-isolated strains of Staphylococcus aureus, limited work has been done concerning these properties against clinical isolates of this pathogen. S. aureus is responsible for an increase number of nosocomial infections and at the same time exhibits increased resistance to synthetic agents.In this study, essential oils from eight aromatic plants common in Greece were isolated by hydrodistillation, analyzed by gas chromatography (GC) and GC/mass spectrometry (GC/MS) for their chemical components and tested for their antimicrobial activities against 24 clinical isolates of S. aureus. The methods used were disk diffusion and broth dilution in order to determine the Minimum Inhibitory Concentration (MIC).Our results showed that essential oils from Origanum vulgare and Origanum dictamnus were active against S. aureus when tested by disk diffusion, but exhibited increased MIC values (>256 mg/L) with the dilution method. In contrast, the reference strain NCTC 6571 showed to be extremely sensitive in most of the oils tested (MICs 0.25−32.0 mg/L) and resistant only to the essential oil from Ocimum basilicum. Therefore, there is no evidence of a potential clinical use for those essential oils and further research is needed in order to determine if they could substitute efficiently synthetic antibiotics or, perhaps be used in combination.     

Evaluation of in vitro antimicrobial activity of leaf and stem essential oils of <Emphasis Type="Italic">Chloroxylon swietenia</Emphasis> DC.

The essential oils obtained by hydrodistillation of leaves and stems of Chloroxylon swietenia DC. were analysed by GC and GC-MS. The main components in the leaf oil were limonene, pregeijerene, geijerene and germacrene D, while stem oil was rich in limonene, methyl eugenol, pregeijerene and geijerene. The essential oils were evaluated for antimicrobial activity against two gram-positive and two gram-negative bacteria and four pathogenic fungi using agar disc diffusion technique. Subsequently, the minimum inhibitory concentration (MIC) from oils was determined by broth microdilution. Both the oils exhibited moderate to strong activities against all the organisms tested. Bacillus subtilis was most susceptible at 100 μg/ml of leaf and stem oils with inhibition zones of 15.9 and 13.1 mm respectively. Among all the fungi tested, A. niger inhibited effectively with a zone of inhibition of 14.9 and 11.5 mm for leaf and stem oils respectively. The results obtained suggest that the essential oils of the plant possess antimicrobial properties and serve as a biofriendly source of antimicrobial ingredients for the food and pharmaceutical industries.     

Analysis of volatile compounds released from embryogenic cultures and somatic embryos of sweet oranges by head space SPME

Volatile compounds released from callus and nucellar embryo tissues of ‘Valencia late’ and ‘Washington Navel’ sweet oranges (Citrus sinensis, L. Osbeck) were collected/concentrated by head space solid phase micro extraction and analysed by gas chromatography-mass spectrometry. Friable, white embryogenic cultures released a number of volatile compounds, including some essential oils. Different samples of the same embryogenic culture showed variability, possibly related to the presence of tissues undergoing differentiation. Analyses of the somatic embryos permitted the identification of several components, including limonene and methyl anthranilate. Considering the simplicity and the very small sample required (0.3 g of fresh tissue) head space solid phase micro extraction is suitable for studies and comparisons of volatile metabolites released from in vitro Citrus tissue cultures suggesting its potential in Citrus biochemical, genetic and breeding research. This revised version was published online in June 2006 with corrections to the Cover Date.     

Fungitoxicity of the essential oil of Citrus sinensis on post-harvest pathogens

Summary The essential oil extracted from the epicarp of Citrus sinensis exhibited absolute fungitoxicity against the 10 post-harvest pathogens. GC–MS studies of the oil revealed the presence of 10 chemical constituents, of which limonene was found to be the major component (84.2%). The activity of the oil was tested by the poisoned food technique (PF) and the volatile activity (VA) assay and the oils showed greater toxicity in the VA assay than in the poisoned food assay. The nature of the toxicity was studied in the VA assay and it was observed that the oil was fungicidal for the 10 pathogens in the 700 ppm (mg/l) to 1000 ppm range. The oil was extremely toxic for spore germination and it was found that at 700 ppm, spore germination was inhibited in the 10 test fungi out of the 12 tested. Treatment at 300 ppm concentration exhibited 70–100% inhibition of spore germination in most of the fungi tested. Scanning electron microscopy (SEM) was done to study the mode of action of the oil in Aspergillus niger and it was observed that treatment with the oil leads to distortion and thinning of the hyphal wall and the reduction in hyphal diameter and absence of conidiophores.     

Antioxidant properties of essential oils from lemon,grapefruit, coriander,clove, and their mixtures

Antioxidant properties of individual essential oils from lemon (Citrus limon L.), pink grapefruit (Citrus paradisi L.), coriander (Coriandrum sativum L.), and clove (Caryophyllus aromaticus L.) buds and their mixtures were studied by capillary gas-liquid chromatography. Antioxidant activity was assessed by oxidation of the aliphatic aldehyde hexanal to the carboxylic acid. The lowest and highest antioxidant activities were exhibited by grapefruit and clove bud essential oils, respectively. Mixtures containing clove bud essential oil also strongly inhibited oxidation of hexanal. Changes in the composition of essential oils and their mixtures in the course of long-term storage in the light were studied. The stability of components of lemon and coriander essential oils in mixtures increased compared to individual essential oils.     

Composition and Antimicrobial Activity of the Essential Oils of Laserpitium latifolium L. and L. ochridanum Micevski (Apiaceae)

The chemical composition and antimicrobial activity of essential oils of Laserpitium latifolium and L. ochridanum were investigated. The essential oils were isolated by steam distillation and characterized by GC‐FID and GC/MS analyses. All essential oils were distinguished by high contents of monoterpenes, and α‐pinene was the most abundant compound in the essential oils of L. latifolium underground parts and fruits (contents of 44.4 and 44.0%, resp.). The fruit essential oil was also rich in sabinene (26.8%). Regarding the L. ochridanum essential oils, the main constituents were limonene in the fruit oil (57.7%) and sabinene in the herb oil (25.9%). The antimicrobial activity of these essential oils as well as that of L. ochridanum underground parts, whose composition was reported previously, was tested by the broth‐microdilution method against four Gram‐positive and three Gram‐negative bacteria and two Candida albicans strains. Except the L. latifolium underground‐parts essential oil, the other investigated oils showed a high antimicrobial potential against Staphylococcus aureus, S. epidermidis, Micrococcus luteus, or Candida albicans (minimal inhibitory concentrations of 13.0–73.0 μg/ml), comparable to or even higher than that of thymol, which was used as reference compound.     

Evaluation of the anti-<Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp <Emphasis Type="Italic">cicer</Emphasis> and anti-<Emphasis Type="Italic">Alternaria porri</Emphasis> effects of some essential oils

The anti-Fusarium oxysporum f. sp cicer (FOC) and anti-Alternaria porri (A. porri) effects were evaluated for 75 different essential oils. The most active essential oils found were those of lemongrass, clove, cinnamon bark, cinnamon leaf, cassia, fennel, basil and evening primrose. However, the effectiveness of these essential oils with both the tested fungi showed different responses. The level of inhibition was compared with Hexaconazole. GC–MS analysis for five oils amongst the 75 essential oils tested was performed. The potential of these essential oils as an ecofriendly and economic approach as a fungicide for FOC and A. porri is discussed.     

In-vitro anti-<Emphasis Type="Italic">Vibrio</Emphasis> spp. activity and chemical composition of some Tunisian aromatic plants

The chemical composition of five aromatic plants (Mentha longifolia, M. pulegium, Eugenia caryophyllata, Thymus vulgaris and Rosmarinus officinalis) frequently used in food preparation in Tunisia was analysed by GC-MS. The antimicrobial effect of the essential oils obtained from these plants was tested against Vibrio alginolyticus, Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio fluvialis strains. Thyme oil exhibited a high level of antimicrobial activities against Vibrio spp. strains. The diameter of the zones of growth inhibition for V. parahaemolyticus species was interestingly high (ranging from 14.66 to 28 mm). The MIC and MBC values were interestingly low for thyme oil (MIC 0.078–0.156 mg/ml) and (MBC >0.31–1.25 mg/ml). These results showed that these plants especially thyme and clove, can be to be used for seafood preparation to protect against contamination by Vibrio spp. strains. An erratum to this article can be found at     

Evaluation of Eupatorium cannabinum Linn. oil in enhancement of shelf life of mango fruits from fungal rotting

Essential oils extracted from 17 higher plants belonging to different families were screened against Botryodiplodia theobromae and Colletotrichum gloeosporioides causing stem end rot disease and anthracnose disease in mango respectively. The essential oil of Eupatorium cannabinum was found to be fungitoxic in nature against both the mango-rotting fungi. Eupatorium oil was standardized through physico-chemical and fungitoxic properties. Gas Liquid Chromatography (GLC) analysis of the oil led to the identification of 16 components, which represented 77.97% of the oil. Germacrene D (16.11%) was found to be the major component. The oil showed a broad fungitoxic spectrum and was recorded to be more efficient than some synthetic fungicides. The oil also showed an inhibitory effect on pectinase and cellulase enzymes. The oil enhanced the shelf life of mango fruits by protecting from fungal rotting when tested as a fumigant. The LD₅₀ of Eupatorium oil was found to be 22.01 ml/kg body weight on mammalian mice.     

Screening for inhibitory activities of essential oils on the growth of Colletotrichum gloeosporioides (Penz.) Penz. & Sacc., the causal agent of leaf spot disease of Murraya koenigii L.

Seven essential oils namely clove, cedar wood, lemongrass, peppermint, eucalyptus, citronella and neem oils were tested for their inhibitory effect on spore germination, growth of germ tube and mycelial growth of Colletotrichum gloeosporioides isolated from diseased Murraya koenigii. All essential oils inhibited the germination and growth of germ tube at different concentrations. However, significant reduction in colony growth was observed with citrus, lemongrass and peppermint oils at 1000, 1500 and 2000 ppm concentrations, respectively. Citrus oil at 1360 ppm inhibited the maximum growth of the fungus followed by lemongrass oil at 1720 ppm and peppermint at 2260 ppm, respectively. The effect of essential oils on mycelial dry weight also showed antifungal activity on the growth of Colletotrichum gloeosporioides. The study revealed the possible utilisation of these essential oils for foliar spray for the management of leaf spot disease of Murraya koenigii.     

Chemical Diversity and Antimicrobial Activity of Volatile Compounds from Zanthoxylum zanthoxyloides Lam. according to Compound Classes,Plant Organs and Senegalese Sample Locations

The chemical diversity of Zanthoxylum zanthoxyloides growing wild in Senegal was studied according to volatile compound classes, plant organs and sample locations. The composition of fruit essential oil was investigated using an original targeted approach based on the combination of gas chromatography (GC) and liquid chromatography (LC) both coupled with mass spectrometry (MS). The volatile composition of Z. zanthoxyloides fruits exhibited relative high amounts of hydrocarbon monoterpenes (24.3 – 55.8%) and non‐terpenic oxygenated compounds (34.5 – 63.1%). The main components were (E)‐β‐ocimene (12.1 – 39%), octyl acetate (11.6 – 21.8%) and decanol (9.7 – 15.4%). The GC and GC/MS profiling of fruit essential oils showed a chemical variability according to geographical locations of plant material. The LC/MS/MS analysis of fruit oils allowed the detection of seven coumarins in trace content. The chemical composition of fruit essential oils was compared with volatile fractions of leaves and barks (root and trunk) from the same plant station. Hexadecanoic acid, germacrene D and decanal were identified as the major constituents of leaves whereas the barks (root and trunk) were dominated by pellitorine (85.8% and 57%, respectively), an atypic linear compound with amide group. The fruit essential oil exhibited interesting antimicrobial activities against Staphylococcus aureus and Candida albicans, particularly the alcohol fraction of the oil.     

23种中草药的体外抗菌活性筛选研究

为考察中草药抗菌物质基础筛选出活性提取物,该研究通过80%乙醇冷浸和95%乙醇回流提取制备23种中草药的提取物,采用琼脂扩散法测量抑菌圈直径,用微量液体培养基倍比稀释法测定最低抑菌浓度(minimum inhibitory concentration,MIC)和最低杀菌浓度(minimum bactericidal/fungicidal concentration,MBC/MFC),并测定了提取物对临床4种常见病原菌体外抗菌活性。结果表明:紫珠草、千斤拔、黄龙尾等9种中草药对金黄色葡萄球菌有较强的抑菌活性,其MIC/MBC值除个别菌是12.5 mg·mL~(-1)外,其他都在0.09～3.12 mg·mL~(-1)之间;千斤拔、大红袍、过江龙等5种中草药对铜绿假单胞菌有较强抑菌活性,其MIC/MBC值在3.12～12.5 mg·mL~(-1)之间;紫珠草、千里光、石楠等13种中草药对大肠埃希菌有较强的抑菌活性,其MIC/MBC值在0.09～6.25 mg·mL~(-1)之间;八角对白色念珠菌有较强抑菌活性,其MIC/MFC值在0.78～12.5 mg·mL~(-1)之间。23种中草药的抗细菌活性较好,尤其是千斤拔、大红袍、过江龙、八角、黄药子对金黄色葡萄球菌、大肠埃希菌、铜绿假单胞菌都具有较好的抑菌活性,具有广谱抗菌活性;但对真菌抑菌效果不明显,仅有八角对白色念珠菌有抑菌活性。此外,提取溶剂浓度、提取温度和提取时间对中草药的提取率和活性均有影响,冷提稍优于热提。     

Investigations on influencing plant-associated bacteria in tissue cultures of black locust (<Emphasis Type="Italic">Robinia pseudoacacia</Emphasis> L.)

During tissue culture of black locust (Robinia pseudoacacia L.), serious problems with plant-associated bacteria led to a reduction of propagation potential in several clones. Four dominant strains of plant-associated bacteria could be isolated and were assigned to the genera Acidovorax, Dyella, Microbacterium and Sphingomonas. Out of five essential oils tested, thyme and lemongrass oil at a concentration of 0.03% each and 0.015% of both oils in combination clearly inhibited the growth of these bacteria strains on bacteriologic medium. There were no significant differences in total bacterial population density when penicillin, thyme and lemongrass oil or thyme plus lemongrass oil were added to the plant propagation media. The use of lemongrass oil changed the proportion of dominant bacterial strains.     

Roots volatiles and fatty acids of coriander (<Emphasis Type="Italic">Coriandrum sativum</Emphasis> L.) grown in saline medium

An hydroponic culture was conducted to investigate the effect of saline stress on the essential oil and fatty acid composition of Tunisian coriander (Coriandrum sativum L.) roots. Ten days old coriander seedlings were treated during 3 weeks with different NaCl concentrations (0, 25, 50 and 75 mM). Roots volatile components and fatty acids were analyzed. The essential oil yield was 0.06% in the control, on the basis of dry matter weight, and did not changed at low concentration (25 mM), while it increased significantly with increasing NaCl concentrations to reach 0.12 and 0.21% at 50 and 75 mM NaCl, respectively. The major volatile component was (E)-2-dodecenal with 52% of total essential oil constituents, followed by decanal, dodecanal, (E)-2-tridecenal and (E)-2-dodecenal. Further, the amount of these compounds was affected differently by the NaCl level. Total fatty acid amount of coriander roots increased significantly only with 50 and 75 mM NaCl. Three major fatty acids: linoleic (43%), oleic (25.5%) and palmitic (21.6%) were identified. Linoleic acid amount remains unchanged at 25 mM, while it increased with raising NaCl concentrations. However, oleic acid amount decreased only at 25 mM and no effect was observed at 50 and 75 mM. Fatty acid percentages were differently affected by salt. The oleic/linoleic ratio was reduced with raising NaCl concentrations.     

Evaluation of commercial essential oil samples on the growth of postharvest pathogen Monilinia fructicola (G. Winter) Honey

Aim: To assess the effect of several commercial essential oils samples Australian lemon myrtle (Backhousia citriodora), cinnamon bark (Cinnamomum zeylanicum), oregano (Origanum vulgare), thyme oil (Thymus vulgaris), clove bud (Eugenia caryophyllata), valerian (Valeriana officinalis) and Australian tea tree oil (Melaleuca alternifolia) on mycelium growth and spore germination of Monilinia fructicola. The effectiveness of lemon myrtle essential oil as a fumigant for the control of brown rot in nectarines was evaluated. Methods and Results: Monilinia fructicola exhibited a different level of sensitivity to each tested essential oil with results suggesting that the essential oils provide excellent control of the pathogen with respect to mycelium growth and spore germination at very low concentrations, whereas for others higher concentrations are needed to reduce significant fungal growth. In vivo application of lemon myrtle essential oil effectively reduced the incidence of M. fructicola on noninoculated fruit. Fumigation of nectarines following inoculation did not reduce the incidence of brown rot in comparison with the inoculated control treatment. No evidence of phytotoxicity on the fruit was recorded. Conclusions: Lemon myrtle essential oil exhibited the strongest antifungal activity against M. fructicola, in vitro and to a lesser extent, under in vivo conditions. Significance and Impact of the Study: The results demonstrate that lemon myrtle essential oil, in particular, has potential as an antifungal agent to control M. fructicola.     

Anti-biofilm properties of Satureja hortensis L. essential oil against periodontal pathogens

Essential oils of several plants are widely used in ethnomedicine for their antimicrobial and anti-inflammatory properties. However, very limited data exist on their use in connection to periodontal diseases. The aim of the present study was to investigate the bacterial growth inhibiting and anti-biofilm effects of Satureja hortensis L. (summer savory), Salvia fruticosa M. (sage), Lavandula stoechas L. (lavender), Myrtus communis L., and Juniperus communis L. (juniper) essential oils. Chemical compositions of the essential oils were analyzed by gas chromatography–mass spectrometry, minimum inhibitor concentrations (MICs) with the agar dilution method, and anti-biofilm effects by the microplate biofilm assay. The toxicity of each essential oil was tested on cultured keratinocytes. Of the 5 essential oils, S. hortensis L. essential oil had the strongest growth inhibition effect. Subinhibitory dose of S. hortensis L. essential oil had anti-biofilm effects only against Prevotella nigrescens. Essential oils did not inhibit keratinocyte viability at the concentrations of 1 and 5 μl/ml, however at the concentration of 5 μl/ml epithelial cells detached from the culture well bottom. The present findings suggest that S. hortensis L. essential oil inhibits the growth of periodontal bacteria in the concentration that is safe on keratinocytes, however, in the subinhibitory concentration its anti-biofilm effect is limited.     

Seasonal variation in allelopathic potential of<Emphasis Type="Italic">Artemisia princeps</Emphasis> var.<Emphasis Type="Italic">orientalis</Emphasis> on plants and microbes

We investigated seasonal variations in allelopathic potential ofArtemisia princeps var.orientalis. Aqueous and meth-anol extracts and volatile substances were prepared in the laboratory from samples collected monthly (April through October). Their impacts were then assessed on the germination and seedling growth ofLactuca sativa andAchyranthes japonica. The allelopathic potential varied with the time of sample collection and the concentration tested. For example, germination ofL. sativa was not inhibited by the aqueous extract but seedling growth (shoots and roots) was, with its seasonal effect being significant. ForA. japonica, seed germination was not inhibited at lower concentrations (except for August samples). However, at higher concentrations and in certain months (especially July), germination was more negatively affected. The degree of seedling growth inhibition also differed by month and by extract concentration, with roots being impacted more than shoots. Volatile substances also had a time-dependent influence on the germination and seedling elongation ofA. japonica. In a separate experiment, the ethyl-acetate and water fractions of a crude methanol extract were prepared monthly fromA. princeps var.orientalis. Here, we examined their antimicrobial activities against three gram-positive bacteria (Bacillus cereus, Bacillus subtilis, andStaphylococcus aureus), two gramnegative bacteria (Escherichia coli andPseudomonas fluorescens), and one lactic acid bacterium,Lactobacillus plantar urn. The ethyl-acetate fraction that was sampled in September was remarkably potent againstB. cereus andB. subtilis, whereas the water fraction collected in August and September showed great antimicrobial activity against the grampositive and -negative bacteria. In contrast,L. plantarum was not inhibited by the water fraction, regardless of the sampling month. Likewise, the ethyl-acetate and water fractions collected in April and October had the lowest levels of antimicrobial activity.     

In vitro inhibition of Candida albicans growth by plant extracts and essential oils

The opportunistic Candida albicans yeast strain ATCC 10261 grows at 37 °C and gives germ tubes after 3 h on corn meal agar and blood plasma. It produces chlamydospores and assimilates sucrose, dextrose, galactose, maltose, trehalose and xylose among the tested carbon sources. Other growth characters were also investigated. The agar diffusion cut plug technique revealed that 200 l of Foeniculum vulgare Miller (fennel), Mentha piperita L. (peppermint) and Citrus limon (lemon) essential oils showed inhibitory actions. Fumigation test of the three essential oils had complete inhibition on the growth. The essential oil of Eucalyptus occidentalis End1 (eucalyptus, eucalypt) had no influence on C. albicans growth by the two tests. Meanwhile, methanol extracts of both leaves and male cones of the conifer Thuja orientalis (eastern thuya) had an inhibitory influence on the growth by two tests (cut plug and filter paper disc assay). In comparative tests the antibiotics mycostatin and dermatin had an inhibitory activity on C. albicans at the concentrations of 50–80 g per hole. Crude methanol extract of leaves of eastern thuya (10–80 g/ml) reduced C. albicans growth and intercellular protein nitrogen on liquid media.