Essential oils against multidrug resistant gram-negative bacteria

Multi-drug resistant uropathogens are responsible for urinary tract infections. The antibacterial activity of seven essential oils, oregano, thyme, clove, arborvitae, cassia, lemongrass, tea tree) was investigated by agar diffusion method, followed by determination of minimum inhibitory (MIC) and bactericidal (MBC) concentrations against five multidrug resistant isolates namely Pseudomonas aeruginosa, Escherichia coli, Enterobacter cloaceae, Morganella morganii, Proteus mirabilis. Oregano, thyme, cassia had antibacterial activity with inhibition zones ranging 25–39 mm; clove, arborvitae, tea tree and lemongrass 12–15 mm. The essential oils showed antibacterial activities with MICs ranged from 0.005% (w/v) to 0.5% (w/v). Thyme had the same MIC and MBC on all strains. The effects of the vapors of the essential oils were also tested by placing the oils on the underside of the Petri dish lid. Thyme, oregano and cassia essential oils strongly inhibited the growth of the clinical strains of bacteria tested in vapor phase. This study demonstrates the potential of investigated essential oils as natural alternatives for further application in hospital therapies in order to retard or inhibit the bacterial growth. For the first time antibacterial effects of essential oils (clove, arborvitae, tea tree, lemongrass, and cassia) were evaluated against Enterobacter cloaceae and Morganella morganii clinical isolates.     

Comparison of solid‐phase cytometry and the plate count method for the evaluation of the survival of bacteria in pharmaceutical oils

Aim: To compare the survival of four bacterial strains (Escherichia coli, Proteus mirabilis, Staphylococcus aureus, Pseudomonas aeruginosa) in pharmaceutical oils, including jojoba oil/tea tree oil, carbol oil, jojoba oil and sesame oil. Methods and Results: Oils were spiked with the test bacteria in a concentration of 10⁴ CFU ml^?1. Bacteria were extracted from oils with phosphate‐buffered saline containing 0·5% Tween 20. Aliquots of the pooled water layers were analysed by solid‐phase cytometry and plate counting. Plate counts dropped to zero for all test strains exposed for 24 h to three of the four oils. In contrast, significant numbers of viable cells were still detected by SPC, except in the jojoba oil/tea tree oil mixture and partly in sesame oil. Conclusions: Exposure of bacteria for 24 h to the two oils containing an antimicrobial led to a loss of their culturability but not necessarily of their viability. The antibacterial activity of the jojoba oil/tea tree oil mixture supersedes that of carbol oil. Significance and Impact of the Study: These in vitro data suggest that the jojoba oil/tea tree oil mixture more than carbol oil inhibits bacterial proliferation when used for intermittent self‐catherization.     

Phytochemical profiling,antioxidant and antibacterial efficacy of a native Himalayan Fern: Woodwardia unigemmata (Makino) Nakai

Present work elucidates the antioxidant and antibacterial activity of Woodwardia unigemmata (Makino) Nakai along with chemical characterization using its aqueous (AEW), methanol (MEW), and hexane (HEW) extracts. Chemical profile of different extracts was illustrated by using Gas chromatography and mass spectrometry (GC-MS) analysis. Antioxidant activities were tested using DPPH and FRAP assays, total phenolic and flavonoid content by Folin-Ciocalteu and aluminum chloride method, respectively. Further, antibacterial activity against six plant and four animal pathogenic bacteria was analyzed by employing the disc diffusion assay. GC-MS analysis revealed the presence of catechol (21.96%), glycerol (20.22%), n-pentadecanoic acid (6.95%), glyceryl monoacetate (6.35 %), ethyl acetimidate (5.39 %) and 3-hydroxy-2,3-dihydromaltol (5.36%) in AEW; β-sitosterol (17.39%), pentadecanoic acid (9.81%), vitamin E (7.82%) and glycerol (7.05%) in MEW; γ-sitosterol (33.45%), vitamin E (10.04%) and campesterol (7.32%) in HEW as major constituents. Maximum phenolics (873 ± 6.01 mgGAE/g dry extract) as well as flavonoids (151 ± 11.44 mgQE/g dry extract) content was found in MEW, which also showed remarkable antioxidant potential (IC₅₀ 6.07 ± 1.4 µg/ml for DPPH and 768 ± 10.4 mg AAE/g dry extract for FRAP assay. In antibacterial activity, maximum inhibition (15 ± 0.9 mm) was observed for HEW against R. solanacearum, followed by AEW against A. tumefaciens and X. phaseoli (11 ± 0.3 mm each). MEW was found positive only against A. tumefaciens. Significant minimum inhibitory concentration (MIC) value observed for AEW against L. monocytogenes (10 mg/ml). Polar extracts had remarkable antioxidant potential, while non-polar extract did show significant antibacterial activity. Further, GC- MS reports indicated that this traditionally useful fern species can be an excellent source of biologically active compounds.     

Larvicidal and adulticidal activity of essential oils from plants of the Lamiaceae family against the West Nile virus vector,Culex pipiens (Diptera: Culicidae)

Culex pipiens mosquitoes are the most widely distributed primary vector of the West Nile virus worldwide. Many attempts for investigation of botanical pesticides to avoid the development of pesticide resistance to conventional synthetic pesticides that are recognized as a threat to the diversity of ecosystems. The study aimed to determine the components of three essential oils of Lamiaceae family, lavender (Lavandula angustifolia), peppermint (Mentha piperita L.), and rosemary (Rosmarinus officinalis L.) by gas chromatography-mass spectrometry (GC–MS) analysis. Furthermore, aimed to validate the insecticidal activities of these oils as larvicidal agents against the third instar larvae of Culex pipiens using five different concentrations (62.5, 125, 250, 500, and 1000 ppm) for each oil in five replicates and as an adulticidal agent against approximately three-day-old female adults of Cx. Pipiens using 0.5, 1, 2, 4, and 5% concentrations in three replicates. The results generally showed a dose-related response. At 1000 ppm, rosemary oil showed the highest larvicidal (100%) (LC₅₀, 214.97 ppm), followed by peppermint oil (92.00% mortality and LC₅₀ (269.35 ppm). Lavender oil showed the lowest efficacy with 87.20% mortality and LC₅₀ (301.11 ppm). At 5% oil concentration, the highest knockdown rate at 1 h was recorded for lavender oil (95.55%), followed by peppermint oil (88.89%) and lastly rosemary oil (84.44%). After 24 h, rosemary oil showed the lowest adult mortality rate (88.89%; LC₅₀, 1.44%), while lavender and peppermint oils both showed a 100% mortality rate, with (LC₅₀, 0.81% and 0.91%, respectively). The chemical constituents of the oils consisted of monoterpenes and sesquiterpenes that determined their insecticidal activities against the target insect stage. The study proposed that rosemary essential oil may be useful for the control of Cx. pipiens larvae as part of an integrated water treatment strategy, and lavender and peppermint oils may be used in an integrated plan for adult’s control.     

Non‐host plant essential oil volatiles with potential for a ‘push‐pull’ strategy to control the tea green leafhopper,Empoasca vitis

The tea green leafhopper, Empoasca vitis Göthe (Hemiptera: Cicadellidae), is an economically important pest of tea crops, Camellia sinensis (L.) O. Kuntze (Theaceae), in China. The use of non‐host plant essential oils for manipulation of E. vitis was investigated for potential incorporation into a ‘push‐pull’ control strategy for this pest. The effectiveness of 14 plant essential oils in repelling E. vitis was investigated in laboratory assays. Rosemary oil, geranium oil, lavender oil, cinnamon oil, and basil oil repelled leafhoppers in a Y‐shaped olfactometer. We also compared the efficacy of these five plant essential oils to repel E. vitis in the presence of a host plant volatile‐based leafhopper attractant, (Z)‐3‐hexenyl acetate, in a tea plantation. In the treatment combination, four plates (north, south, east, and west) treated with an essential oil surrounded a central sticky plate treated with (Z)‐3‐hexenyl acetate. Fewer E. vitis were found on the plates treated with rosemary oil (12.5% reduction) than on the four water‐sprayed control treatment plates surrounding a central plate with (Z)‐3‐hexenyl acetate. We compared the distribution of E. vitis on the plates, and the relative numbers of E. vitis on each plate were compared with similar plates in the control treatment. When four plates treated with rosemary oil surrounded a central (Z)‐3‐hexenyl acetate‐treated plate, the distribution of E. vitis on the different plates changed significantly compared with that of the control. Relatively fewer E. vitis were found on the east (13.0% reduction) rosemary oil‐treated plates and more E. vitis (11.3% increase) were found on the central attractant‐treated plate. Our findings indicate that rosemary oil is a promising leafhopper repellent that should be tested further in a ‘push‐pull’ strategy for control of E. vitis.     

栽培地区、采收季节和株龄对迷迭香精油成分和抑菌活性的影响

采用气相色谱-质谱联用的方法,研究了不同提取部位、栽培地区、采收季节和株龄对迷迭香精油提取率、成分和抑菌活性的影响。结果表明,在贵州栽培的迷迭香叶子中精油的主要成分为α-蒎烯（31.88%–39.09%）、1,8-桉叶素（8.82%–10.97%）和樟脑（7.63%–11.59%）;茎中精油的主要成分为龙脑（26.62%–39.05%）、樟脑（11.41%–15.27%）和α-蒎烯（7.23%–17.80%）;茎中精油的抑菌活性强于叶中精油。北京地区栽培的迷迭香,其叶子中精油的主要成分为樟脑（27.92%）和L-β-蒎烯（25.28%）,抑菌活性强;茎中精油和贵州栽培的成分和活性均相似。对于采收季节,夏季采收的迷迭香精油提取率高,抑菌活性强。而株龄对精油的成分和抑菌活性影响不大。该研究为我国引种迷迭香的合理配置栽培、采收、生产条件和高效开发利用其抑菌活性提供了科学的依据。     

几种典型植物精油的化学成分与其抗菌活性

【目的】植物精油萃取自天然植物, 因具有抗菌活性, 近年来受到广泛关注。论文的目的是分析植物精油的化学成分, 测试其抗菌活性, 并研究其化学成分与抗菌活性之间的联系。【方法】实验选取了肉桂、山苍子、丁香、香茅、迷迭香和大蒜精油等6种典型植物精油, 通过气质联用分析方法研究了其化学组分, 并通过污染食物技术研究了其对黑曲霉和绳状青霉的抗真菌活性, 以及对大肠杆菌和金黄色葡萄球菌的抗细菌活性。【结果】气质联用分析结果表明, 肉桂、山苍子、香茅和迷迭香等4种植物精油的化学成分主要是醛类和醇类, 丁香精油的主要化学成分是丁香油酚, 大蒜精油化学成分基本上都是含硫的醚类, 其中二烯丙基三硫醚(大蒜素)含量最高。抗菌活性结果显示, 不同植物精油的抗菌活性不同, 6种植物精油的抗真菌活性由强到弱依次为: 肉桂>大蒜>丁香>山苍子=香茅>迷迭香, 抗细菌活性由强到弱依次为: 肉桂>山苍子>丁香>香茅=迷迭香>大蒜。【结论】植物精油的抗真菌、细菌活性与其化学组分密切相关, 肉桂、山苍子、香茅和迷迭香等4种精油的抗菌活性可能主要与其化学成分中的醛类和醇类有关, 丁香精油较高的抗菌活性可能主要源于丁香油酚; 大蒜精油具有高效的抗真菌活性主要源于其化学成分中的含硫醚。不同植物精油化学成分不同, 抗真菌、细菌活性也不同, 表明其可能有不同的抗真菌与抗细菌机制。     

Chenopodium album extract ameliorates carbon tetrachloride induced hepatotoxicity in rat model

Major objective of this study was to explore the protective effect of the methanolic extract of Chenopodium album against carbon tetrachloride induced hepatotoxicity in rats. Chenopodium album has locally been used for multiple medicinal proposes. Methanolic extract of Chenopodium album (whole plant) was prepared with Soxhlet extractor and rotatory evaporator. Antioxidant activity of Chenopodium album was determined by DPPH free radical scavenging assay. Thirty Wister (albino) rats (150–200 g) were divided into six groups for the evaluation of hepatoprotective potential of different concentrations of Chenopodium album against carbon tetrachloride (1:1 CCl₄: Olive oil) under the controlled laboratory conditions. Group-I rats were administrated with olive oil (Normal control), Group-II with CCl₄ only, Group-III with Silymarin (positive control), Group-IV with Chenopodium album (100 mg/kg), Group-V with Chenopodium album (200 mg/kg) and Group-VI rats with Chenopodium album (300 mg/kg) for the period of 28 days. Serum was taken to determine the levels of alanine transaminase, aspartate transaminase, alkaline phosphatase, cholesterol, triglyceride, creatinine and urea in the blood. Formalin stored tissues were examined for histopathological analysis. DPPH assay showed that Chenopodium album has the potential for reduction of oxidative stress. Chenopodium album minimized the levels of ALT (70 ± 8.68 U/L, 68.75 ± 8.38 U/L & 73.5 ± 10.28 U/L), AST (219.5 ± 19.16 U/L, 140.75 ± 13.35 U/L & 221.25 ± 13.33 U/L) and ALP (289.5 ± 28.21 U/L, 258 ± 11.12 U/L & 248.25 ± 4.03 U/L) at different concentrations (100 mg/kg, 200 mg/kg, 300 mg/kg respectively). Chenopodium album enhanced triglyceride level (64.75 ± 12.66 mg/dl at 200 mg/kg) as compared to CCl₄ treated group (33.25 ± 1.26 mg/dl). Carbon tetrachloride elevated urea level (43.25 ± 6.6) was decreased by high dose of Chenopodium album (18 ± 8.17). Moreover, Chenopodium album also improved WBC level (9.69 × 10³ /Cu.mr & 10.59 × 10³ /Cu.mr at low and medium doses respectively), RBCs level (6.97 × 10³ /Cu.mr) and hemoglobin level (13.95 G/dL, 13.467 G/dL & 14.05 G/dL at low, medium and high doses). In vivo study of Chenopodium album methanolic extract demonstrates the potential for protection of liver and after pre-clinical studies the plant can be used as a safe alternative of commercially available hepatoprotective medicines.     

Chemical composition and anti-Helicobacter pylori effect of Satureja bachtiarica Bunge essential oil

Resistance of H. pylori strains to common antibiotics has been developed in different parts of the world and continues to increase. It is important to investigate the novel and efficient anti-H. pylori drugs, among which the plants would be suitable sources.Satureja bachtiarica Bunge is traditionally used as antimicrobial agent. In this study, we evaluated the antibacterial activity of S. bachtiarica Bunge essential oil against 10 clinical isolates of Helicobacter pylori by disc diffusion and agar dilution methods. The chemical composition of essential oil was analyzed by GC and GC–MS. Carvacrol (45.5%) and thymol (27.9%) were the primary constituents of oil, followed by p-cymene (4.4%), and γ-terpinene (4.0%). S. bachtiarica essential oil showed strong antibacterial activity against clinical isolates of H. pylori (17.6 ± 1.1 mm and 0.035 ± 0.13 μl/ml). Carvacrol, as the first main component, had a significant role in this effect, whereas in the presence of thymol, the antibacterial effect of carvacrol was reduced. Therefore, S. bachtiarica essential oil can be applied as an alternative agent for treatment of H. pylori infections. More studies would be required to better clarify its mechanism of action on H. pylori.     

Potential effect of Allium sativum bulb for the treatment of biofilm forming clinical pathogens recovered from periodontal and dental caries

Biofilm producing clinical bacterial isolates were isolated from periodontal and dental caries samples and identified as, Lactobacillus acidophilus, Streptococcus sanguis, S. salivarius, S. mutansand Staphylococcus aureus. Among the identified bacterial species, S. aureus and S. mutansshowed strong biofilm producing capacity. The other isolated bacteria, Streptococcus sanguis, S. salivarius showed moderate biofilm formation. These pathogens were subjected for the production of extracellular polysaccharides (EPS) in nutrient broth medium and the strain S. aureus synthesized more amounts of EPS (610 ± 11.2 µg/ml) than S. sanguis (480 ± 5.8 µg/ml).EPS production was found to be less in S. salivarius (52 ± 3.8 µg/ml).The solvent extract of A. sativum bulb showed the phytochemicals such as, carbohydrate, total protein, alkaloids, saponins, flavonoids, tannins and sterioids. The solvent extract of A. sativum bulb showed wide ranges of activity against the selected dental pathogens. The difference in antibacterial activity of the solvent extract revealed differences in solubility of phytochemicals in organic solvents. Ethanol extract was highly active againstS. aureus (25 ± 2 mm). The Minimum Inhibitory Concentration (MIC) of crude garlic bulb varied widely and this clearly showed that bacteria exhibits different level of susceptibility to secondary metabolites. MIC value ranged between 20 ± 2 mg/ml and 120 ± 6 mg/ml and Minimum Bactericidal Concentration (MBC) value ranged from 60 ± 5 mg/l to 215 ± 7 mg/ml. To conclude, A. sativum bulb can be effectively used to treat periodontal and dental caries infections.     

Antimicrobial potential of Indian Cinnamomum species

Cinnamomum is the largest genus of Lauraceae family and has been used as spices, food, and food additives by the people. Total 15 Cinnamomum species are distributed in different parts of Indian sub-continent. Different parts (leaves, stem bark, stem wood, roots, flowers, and fruits) of these species were shade-dried and used for the determination of essential oils. A total of 19 essential oils were identified and quantified from the different parts of (leaf, stem bark, stem wood, root, flower, and fruit) of 15 Cinnamomum species. The stem bark of C. altissimum was rich in the presence of essential oils (52.2 %) whereas minimum levels of essential oils were recorded in roots (17.9 %). The γ-terpinene (11.1 %) was reported as the major component essential oil in C. subavenium flowers. Methanol extract of C. camphora stem wood showed stronger lowest minimum inhibitory concentration against S. aureus (25 ± 0.01 μg/ml), H. pylori (29 ± 0.05 μg/ml), B. subtilis (31 ± 0.03 μg/ml), E. faecalis (33 ± 0.01 μg/ml), C. albicans (38 ± 0.03 μg/ml) when compared to amoxycillin (S. aureus 56 ± 0.05 μg/ml; B. subtilis 27 ± 0.04 μg/ml, E. faecalis 22 ± 0.01 μg/ml), streptomycin (H. pylori 38 ± 0.02 μg/ml) and fluconazole (C. albicans 56 ± 0.01 μg/ml). Methanolic extract of C. camphora stem wood demonstrated maximum antimicrobial activity against S. aureus, H. pylori, B. subtilis, E. faecalis and C. albicans. The essential oil of C. altissimum stem bark displayed significant lowest MIC against S. aureus (21 ± 0.03 μg/ml), E. coli (22 ± 0.03 μg/ml), E. cloacae (37 ± 0.06 μg/ml), L. monocytogenes (47 ± 0.08 μg/ml), and P. chrysogenum (101 ± 0.07 μg/ml) when compared to amoxycillin (E. coli 18 ± 0.01 μg/ml, E. cloacae 21 ± 0.05 μg/ml, L. monocytogenes 31 ± 0.03 μg/ml), and fluconazole (P. chrysogenum 101 ± 0.07 μg/ml). The essential oil of C. altissimum stem bark displayed maximum antimicrobial activity against S. aureus, E. coli, E. cloacae, L. monocytogenes, and P. chrysogenum. Cinnamomum essential oils may be used as an alternative source of antibacterial and antifungal compounds in the treatment of various types of infections.     

Novel strategies of essential oils,chitosan, and nano- chitosan for inhibition of multi-drug resistant: E. coli O157:H7 and Listeria monocytogenes

Despite the wide range of available antibiotics, food borne bacteria demonstrate a huge spectrum of resistance. The current study aims to use natural components such as essential oils (EOs), chitosan, and nano-chitosan that have very influential antibacterial properties with novel technologies like chitosan solution/film loaded with EOs against multi-drug resistant bacteria. Two strains of Escherichia coli O157:H7 and three strains of Listeria monocytogenes were used to estimate antibiotics resistance. Ten EOs and their mixture, chitosan, nano-chitosan, chitosan plus EO solutions, and biodegradable chitosan film enriched with EOs were tested as antibacterial agents against pathogenic bacterial strains. Results showed that E. coli O157:H7 51,659 and L. monocytogenes 19,116 relatively exhibited considerable resistance to more than one single antibiotic. Turmeric, cumin, pepper black, and marjoram did not show any inhibition zone against L. monocytogenes; Whereas, clove, thyme, cinnamon, and garlic EOs exhibited high antibacterial activity against L. monocytogenes with minimum inhibitory concentration (MIC) of 250–400 μl 100^?1 ml and against E. coli O157:H7 with an MIC of 350–500 μl 100^?1 ml, respectively. Among combinations, clove, and thyme EOs showed the highest antibacterial activity against E. coli O157:H7 with MIC of 170 μl 100^?1 ml, and the combination of cinnamon and clove EOs showed the strongest antibacterial activity against L. monocytogenes with an MIC of 120 μl 100^?1 ml. Both chitosan and nano-chitosan showed a promising potential as an antibacterial agent against pathogenic bacteria as their MICs were relatively lower against L. monocytogenes than for E. coli O157:H7. Chitosan combined with each of cinnamon, clove, and thyme oil have a more effective antibacterial activity against L. monocytogenes and E. coli O157:H7 than the mixture of oils alone. Furthermore, the use of either chitosan solution or biodegradable chitosan film loaded with a combination of clove and thyme EOs had the strongest antibacterial activity against L. monocytogenes and E. coli O157:H7. However, chitosan film without EOs did not exhibit an inhibition zone against the tested bacterial strains.     

Loranthus acaciae: Alternative medicine for β-lactamase producer and methicillin-resistant Staphylococcus aureus

Recently, we reported high antibacterial efficiency of Loranthus acaciae (LA) against different standard strains of bacteria including Methicillin-Resistant Staphylococcus aureus (MRSA). Therefore, this study aimed to confirm the effectiveness of LA against clinically isolated Staphylococcus aureus (SA) including β-lactamase producer (Blac) and MRSA. Forty-eight SA isolates collected from various clinical samples were used in this study. Antibiotics susceptibility profile was determined for twenty different antibiotics using automated Microscan Walkaway 96 Plus system as recommended by Clinical and Laboratory Standards Institute (CLSI) guidelines. This system also identified β-lactamase producers and MRSA. In the meantime, LA ethanolic extract was fractionated using liquid–liquid fraction method to hexane, dichloromethane DCM and methanol 80% fractions. Antimicrobial activities of LA extract and fraction were performed with agar well diffusion method for all SA isolates, MIC and MBC were also recorded. Phytochemical screening for various phyto-constituent classes of LA ethanolic extract was determined. Out of 48 SA isolates, Cefoxitin-positive MRSA represent 31 (64.6%), Blac 17 (35.4%), and 41 (85.4%) were multidrug-resistant SA, which was resistant at least to one antibiotic from three different categories. All isolates were resistant to ampicillin and penicillin. Antimicrobial activities of LA extract and fractions revealed that ethanol extract was active against all isolated SA with inhibition zone ranged from 33 ± 2.00 to 25 ± 3.05 mm. While DCM exhibited the largest inhibition zone range from 37 ± 3.00 to 33 ± 2.00 mm. This study is first of its kind conforming the high antibacterial activity of LA against SA isolated from a different source of infection. The study concluded that LA extract and fractions are active and give positive result for all isolated SA. Therefore, suitable pharmacological formulation of LA extract as a promising antibacterial agent for the treatment of SA infection should be given extreme priority.     

Biological activity of Cymbopogon schoenanthus essential oil

IntroductionA number of plant species, including Cymbopogon schoenanthus, are traditionally used for the treatment of various diseases. C. schoenanthus is currently, traded in the Saudi markets, and thought to have medicinal value. This study aimed at investigating the biological activities of C. schoenanthus against both Gram-positive and Gram-negative bacteria and to identify its chemical ingredients.Materials and methodsThe inhibitory effects of water extracts of C. schoenanthus essential oils were evaluated against ten isolates of both Gram-positive and Gram-negative bacteria using the agar well diffusion and dilution methods. The minimum inhibitory concentration (MIC) was assayed using the Broth microdilution test on five of the ten isolates. The death rates were determined by the time kill assay, done according to the Clinical Laboratory Standards Institute (CLSI) guidelines. The chemical composition of the essential oils of the plant was performed using GC/MS.ResultsThe C. schoenanthus essential oil was effective against Escherichia coli, Staphylococcus aureus, methicillin-sensitive (MSSA) S. aureus (MRSA) and Klebsiella pneumoniae. The essential oil was not effective against Staphylococcus saprophyticus at the highest concentration applied of >150 μg/ml. The MIC values were as follows: 9.37 μg/ml for E. coli 4.69 μg/ml for S. aureus (MRSA), 2.34 mg/ml for MSSA and 2.34 μg/ml for K. pneumoniae. The time-kill assay indicated that there was a sharp time dependent decline in K. pneumoniae counts in the presence of the oil. This is in contrast to a gradual decline in the case of S. aureus under the same conditions. The eight major components of the essential oil were: piperitone (14.6%), cyclohexanemethanol (11.6%), β-elemene (11.6%), α-eudesmol (11.5%), elemol (10.8%), β-eudesmol (8.5%), 2-naphthalenemethanol (7.1%) and γ-eudesmol (4.2%).ConclusionThe results of the present study provide a scientific validation for the traditional use of C. schoenanthus as an antibacterial agent. Future work is needed to investigate and explore its application in the environmental and medical fields. In addition, to evaluating the efficacy of the individual ingredients separately to better understand the underlying mechanism.     

In vitro evaluation of the effects of some plant essential oils on Ascosphaera apis,the causative agent of Chalkbrood disease

Ascosphaera apis is one of the major fungal pathogens of honey bee broods and the causative agent of Chalkbrood disease. The factors responsible for the pathogenesis of Chalkbrood disease are still not fully understood, and the increasing resistance of A. apis to commonly used antifungal agents necessitates a search for new agents to control this disease. The in vitro antifungal activities of 27 plant essential oils against two isolates of A. apis (Aksu-4 and Aksu-9) were evaluated. Out of the 27 plant essential oils tested, 21 were found to be effective in killing both isolates of A. apis. Based on their minimum fungicidal concentration (MFC) values, the effective oils were grouped into three categories: highly effective, moderately effective and minimally effective. Mountain pepper oil, Kala Bhangra oil, spearmint oil, babuna oil, betel leaf oil, carrot seed oil, cumin seed oil and clove bud oil were highly effective, with MBC values between 50.0 μg/mL and 600.0 μg/mL. Mountain pepper was the most effective essential oil, with an MBC value of 50.0 μg/mL. Citral and caryophyllene containing oils were the most effective with MIC 50 ppm. The essential oils tested exhibited significant antimicrobial activities against both strains of A. apis, and they may contain compounds that could play an important role in the treatment or prevention of Chalkbrood disease of honeybee.     

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.     

Synergistic effect of Myrtus communis L. essential oils and conventional antibiotics against multi-drug resistant Acinetobacter baumannii wound isolates

Acinetobacter baumannii is a rapidly emerging, highly resistant clinical pathogen with increasing prevalence. In recent years, the limited number of antimicrobial agents available for treatment of infections with multi-drug resistant (MDR) strains reinforced tendency for discovery of novel antimicrobial agents or treatment strategies. The aim of the study was to determine antimicrobial effectiveness of three Myrtus communis L. essential oils, both alone and in combination with conventional antibiotics, against MDR A. baumannii wound isolates. The results obtained highlighted the occurrence of good antibacterial effect of myrtle oils when administered alone. Using checkerboard method, the combinations of subinhibitory concentrations of myrtle essential oils and conventional antibiotics, i.e. polymixin B and ciprofloxacine were examined. The results proved synergism among M. communis L. essential oils and both antibiotics against MDR A. baumannii wound isolates, with a FIC index under or equal 0.50. Combination of subinhibitory concentrations of essential oils and ciprofloxacin most frequently reduced bacterial growth in synergistic manner. The similar has been shown for combination with polymyxin B; furthermore, the myrtle essential oil resulted in re-sensitization of the MDR wound isolates, i.e. MICs used in combination were below the cut off for the sensitivity to the antibiotic. Time-kill curve method confirmed efficacy of myrtle essential oil and polymyxin B combination, with complete reduction of bacterial count after 6 h. The detected synergy offers an opportunity for future development of treatment strategies for potentially lethal wound infections caused by MDR A. baumannii.     

迷迭香油和花椒油对朱砂叶螨的生物活性

【目的】明确室内条件下迷迭香油和花椒油对朱砂叶螨Tetranychus cinnabarinus (Boisduval)的生物活性。【方法】采用熏蒸法和驱避法测定迷迭香油和花椒油对5日龄朱砂叶螨雌成螨的熏蒸活性、产卵抑制活性和驱避活性;并采用EthoVision XT6分析亚致死浓度下两种精油对雌成螨行为的影响。【结果】分别将5日龄雌成螨熏蒸处理12, 24和48 h后,迷迭香油对朱砂叶螨的致死率均显著高于花椒油处理组（P＜0.05）,且随着熏蒸时间的延长或精油浓度的升高,其熏蒸活性均增强;不同浓度的迷迭香油处理朱砂叶螨雌成螨24 h后的产卵抑制率均显著高于花椒油处理组（P＜0.05）。迷迭香油和花椒油处理朱砂叶螨雌成螨24 h的LC₃₀分别为6.731和120.142 μL/L;迷迭香油对朱砂叶螨雌成螨的驱避活性明显高于花椒油处理组。两种精油亚致死浓度处理朱砂叶螨雌成螨后,其自发运动方式发生了明显变化,移动频率在LC₁₀ 时呈现最高;且迷迭香油对朱砂叶螨雌成螨自发运动能力的影响比花椒油处理的影响更为明显。【结论】迷迭香油对朱砂叶螨的生物活性明显高于花椒油,因此,迷迭香油对朱砂叶螨的生物防治具有较高的应用价值。本研究为绿色蔬菜植物源杀螨剂的研究提供了科学依据。     

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.     

Additive and synergistic effects of Bacillus spp. isolates and essential oils on the control of phytopathogenic and saprophytic fungi from medicinal plants and marigold seeds

Contamination of plants and seeds with microorganisms is one of the main problems in the production and distribution of various agricultural products, as well as raw herbal material for the preparation of herbal remedies. In targeting microbial contamination, among other bacteria, Bacillus species showed a significant capacity for biocontrol. The antifungal activity of 14 isolates of Bacillus spp. against 15 fungal isolates from medicinal plants was examined utilizing a dual plate assay. The strongest and broadest antagonistic activity against all fungi tested was exhibited by isolates SS-12.6 and SS-13.1 (from a 43% to 74% reduction in fungal growth), while isolates SS-39.1 and SS-39.3 were effective against the fewest fungus species and also had the weakest antifungal activity. The effect of a crude lipopeptide extract (CLE) of Bacillus sp. SS-12.6 was similar to that achieved by a dual culture with isolate SS-12.6, confirming that the antagonism was the result of the antifungal activities of lipopeptides. In addition, essential oils of thyme (0.55 mg/mL) and savory (0.32 mg/mL) in various combinations with the CLE of SS-12.6 were tested for antifungal activity, and additive and synergistic effects for some of the fungi were obtained. When testing the effect of CLE, oils (0.40 mg/mL for thyme oil and 0.21 mg/mL for savory oil) and combinations in situ on marigold seeds, a reduction of total fungal infection without an adverse effect on germination was accomplished by 6-h treatments with CLE of SS-12.6 (85% reduction of fungal infection and 63% germination), supernatant from liquid culture of SS-12.6 (more than 90% reduction of fungal infection with 69% seed germination) and combinations of CLE and savory oil (77% reduction of fungal infection and 62% seed germination) and CLE with thyme and savory oils (about 75% reduction of fungal infection with 69% seed germination).