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.

Acaricidal activity of Foeniculum vulgare against Rhipicephalus annulatus is mainly dependent on its constituent from trans-anethone.

Globally, the economic losses due to hard ticks infestation and the control of the associated diseases have been calculated at USD $13.9-18.7 billion per year. The economic impact is related to its direct damage to the skins, blood loss, anemia, severe immunological reactions and indirect losses that related to the effects of hemoparasites, cost of treatment for clinical cases and expenses incurred in the control of ticks. The current study evaluated the acaricidal activities of fennel Foeniculum vulgare essential oil and its main components; trans-anethole and fenchone; against R. annulatus. GC-MS analysis revealed that this oil contained 16 components representing 99.9% of the total identified compounds with E-anethole being the predominant component(64.29%), followed by fenchone (9.94%). The fennel oil and trans-anethole showed significant acaricidal activities. The LC50 of the fennel oil was attained at concentrations of 12.96% for adult ticks and 1.75% for tick larvae meanwhile the LC50 of trans-anethole was reached at concentrations of 2.36% for adult tick and 0.56% for tick larvae. On the contrary, fenchone showed no any significant adulticidal activities and its LC50 attained at a concentration of 9.11% for tick larvae. Regarding repellence activities, trans-anethole achieved 100% repellency at the concentration of 10% while fennel showed 86% repellency at the same concentration. Fenchone showed no repellency effect. Treatment of larvae with fennel, trans-anethole, and fenchone LC50 concentrations significantly inhibited the acetylcholinesterase activity. Meanwhile, glutathione s-transferase activity was significantly decreased in fennel treated larvae but no significant effect was found in the larvae of trans-anethole and fenchone groups. These results indicate that the acaricide effect of fennel oil may attributed to its high content of trans-anethole. This was supported by potent adulticidal, larvicidal, and repellency effects of trans-anethole against Rhipeciphalus annulatus tick and therefore it could be included in the list of acaricide of plant origin.

Effect of cytokinins on in vitro multiplication, volatiles composition and rosmarinic acid content of Thymus leucotrichus Hal. shoots.

An efficient in vitro multiplication protocol was designed to Thymus leucotrichus, a subshrub and perennial herb growing naturally in the Northwest of Turkey. Of all basal media studied, Murashige and Skoog medium was found to be superior to the others, providing higher shoot formation and the maximum shoot length. Varying concentrations of cytokinins, i.e., 6-benzyladenine, thidiazuron, 2-isopentenyladenine and kinetin were supplemented in the nutrient media to observe their effects on shoot development and biomass. Rosmarinic acid content and volatile compositions of both naturally growing plants and in vitro multiplied plantlets were also evaluated. 6-benzyladenine (1.0 mg/L) and kinetin (0.5 mg/L) were found to be optimum for shoot number and shoot elongation, respectively. Thidiazuron (1.0 mg/L) was superior for biomass production. Rosmarinic acid content of in vitro multiplied plants was found to be higher than that of wild plants, reaching a maximum with 0.5 mg/L 2-isopentenyladenine, which yielded 10.15 mg/g dry weight. The highest thymol content was obtained with 1.0 mg/L kinetin (55.82%), while thidiazuron (0.1 mg/L) increased carvacrol production (12.53%). Overall, Murashige and Skoog medium supplemented with 1.0 mg/L kinetin was determined to be the most favorable medium studied.

Phenolic acid contents, essential oil compositions and antioxidant activities of two varieties of Salvia euphratica from Turkey.

This study was designed to evaluate the in vitro antioxidant activities of n-hexane (Hex), dichloromethane (DCM), methanol (MeOH) and essential oils (EO) extracts obtained from Salvia euphratica var. euphratica and Salvia euphratica var. leiocalycina and to determine their essential oil and phenolic acid compositions. The samples were screened for their antioxidant activity by using DPPH and ß-carotene/linoleic acid assays. Methanol extracts of both varieties exhibited strong antioxidant activities. Our results showed that rosmarinic acid was dominant phenolic acid of MeOH extracts (39.4 and 55.8 µg mg⁻¹, respectively). The chemical compositions of essential oils of two varieties were analysed and their main components were determined as eucalyptol (18.4%) and trans-pinocarvyl acetate (24.9%), respectively. It can be said that these varieties could be used as natural antioxidant.

Wound healing effects of arnebia densiflora root extracts on rat palatal mucosa.

OBJECTIVES: The aim of this study was to investigate wound healing effects of Arnebia densiflora (Nordm.) Ledeb. root extracts on rat palatal mucosa. METHODS: A 10-mm full-thickness mucosal wounds were created on midline of rats' palate by using scalpel. In the experimental groups, a ten percent extract of A. densiflora roots was topically applied once a day as ointment on the wounds. After wounding, tissue samples from palatal mucosa were harvested at 4, 7, 14 and 21 days and then evaluated histologically. RESULTS: It was observed that 10% A. densiflora root extract has progressive effects on wound healing in experimental groups. CONCLUSIONS: This study suggests that A. densiflora root extract could be developed as a therapeutic agent for wound healing.

Antimicrobial and antioxidant activity of the essential oil and methanol extract of Nepeta cataria.

Catnip (Nepeta cataria) is an important medicinal herb belonging to the mint family, Lamiaceae. In this study, the in vitro antimicrobial and antioxidant activities of the essential oil and methanol extract from Nepeta cataria, and its essential oil composition were investigated. The essential oil, which has 4aalpha,7alpha,7abeta-nepetalactone (70.4%), 4aalpha,7alpha,7abeta-nepetalactone (6.0%), thymol (2.3%), and 4aalpha,7alpha, 7abeta3-nepetalactone (2.5%), as main components, exhibited activity against eleven bacteria, and twelve fungi and a yeast, C. albicans; with Minimum Inhibitory Concentrations (MIC) values ranging from 12.50 to 250 microl/ml; the methanol extract showed weaker activity. The samples were also subjected to a screening for their possible antioxidant activities by using 2.2-diphenyl-1-picrylhydrazyl (DPPH) and beta-carotene/linoleic acid assays. In DPPH assay, the extract showed slight antioxidant activity whereas the essential oil remained inactive. In the latter case, both the extract and the essential oil exerted weak activity having inhibiton ratios of linoleic acid oxidation at 16.4% and 27.0%, respectively. The weak antioxidative nature of the extract could be attributed to the low phenolic content, estimated as gallic acid equivalent at 22.6 +/- 2.07 microg/ml or 2.26%. In both systems, antioxidant capacity of BHT was determined in parallel experiments.

Production and optimisation of rosmarinic acid by Satureja hortensis L. callus cultures.

In this study, production and optimisation of rosmarinic acid, a phenolic acid and an economically important metabolite, was investigated in the callus cultures established from the mature seeds of Satureja hortensis L. (summer savory) plant. Gamborg's B5 basal medium, supplemented with indol butyric acid (IBA) (1.00 mg L(-1)), N6-benzyl aminopurine (6-BA) (1.00 mg L(-1)) and sucrose (2.5%, w/v), was employed for the establishment and maintenance of the callus cultures. Applications were individually prepared by preparing the media containing different IBA/6-BA combinations and sucrose concentrations. All of the applications were carried out in the continuous dark. In the applications, where the effects of IBA/6-BA combinations on the growth and rosmarinic acid accumulation were assayed (1-15 applications), the highest biomass yield was obtained from the medium supplemented with 1.00 mg L(-1) IBA and 5.00 mg L(-1) 6-BA. In the case of the rosmarinic acid accumulation, an opposite relationship was determined between the growth and rosmarinic acid production. While the highest biomass yield was obtained from the medium containing 1.00 mg L(-1) IBA and 5.00 mg L(-1) 6-BA, the highest rosmarinic acid accumulation was obtained from the medium supported with 1.00 mg L(-1) IBA and 1.00 mg L(-1) 6-BA. In the applications where the effects of sucrose concentrations on the growth and rosmarinic acid accumulation were examined, the highest biomass yield was obtained from the medium which is supplemented with 5.0% (w/v) sucrose. In this category, the highest rosmarinic acid accumulation was obtained from the medium which is supported with 3.0% (w/v) sucrose. According to the experiments carried out with the wild S. hortensis, it is found to have 25.02+/-1.21 mg g(-1) rosmarinic acid. No differentiation was observed in any callus during the course of this study.

Chemical composition and antimicrobial and antioxidant activities of the essential oil and methanol extract of Hippomarathrum microcarpum (Bieb.) from Turkey.

Hippomarathrum microcarpum grows wild in eastern Anatolia, Turkey, and is a plant utilized as food by people. In this study, the in vitro antimicrobial and antioxidant activities of the essential oil and methanol extract from H. microcarpum and its essential oil composition were investigated. The essential oil, which has bornyl acetate, caryophyllene oxide, and beta-caryophyllene as its main components, exhibited activity against eight bacteria, nine fungi, and a yeast, Candida albicans, with minimum inhibitory concentration values ranging from 62.50 to 125 muL/mL; the methanol extract showed weak activity. The antioxidant activity of these extracts was assessed by the beta-carotene bleaching test and the 1,1'-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging test. The inhibition of linoleic acid oxidation was very weak for both extracts tested. The inhibition percentages were found to be 22.9 and 33.5% for methanol and essential oil, respectively, at the concentration of 2 g/L. The oil scavenged DPPH at higher concentrations (IC50 = 10.69 +/- 0.05 mg/mL), but the methanol extract exhibited no activity. The total phenolic content of the methanol extract was found to be 4.7 +/- 0.1%.

Screening of the antioxidative properties and total phenolic contents of three endemic Tanacetum subspecies from Turkish flora.

Methanolic extracts of three different Tanacetum subspecies [Tanacetum densum (Lab.) Schultz Bip. subsp. sivasicum Hub-Mor and Grierson, Tanacetum densum (Lab.) Schultz Bip. subsp. eginense Heywood and Tanacetum densum (Lab.) Schultz Bip. subsp. amani Heywood] which are endemic to Turkish flora were screened for their possible antioxidant activities by two complementary test systems namely DPPH free radical scavenging and beta-carotene/linoleic acid. In DPPH system, the most active plant was T. densum subsp. amani with an IC(50) value of 69.30+/-0.37 microg/ml. On the other hand, T. densum subsp. sivasicum exerted greater antioxidant activity than those of other subspecies in beta-carotene/linoleic acid system (79.10%+/-1.83). Antioxidant activities of BHT, curcumine and ascorbic acid were also determined as positive controls in parallel experiments. Total phenolic constituents of the extracts of Tanacetum subspecies were performed employing the literature methods involving Folin-Ciocalteu reagent and gallic acid as standard. The amount of total phenolics was highest in subsp. sivasicum (162.33+/-3.57 microg/mg), followed by subsp. amani (158.44+/-2.17 microg/mg). Especially, a positive correlation was observed between total phenolic content and antioxidant activity of the extracts.

[Can garlic (Allium sativum) extract be used as scolocidal agent?]. / Sarimsak (Allium sativum) özütü skolosidal ajan olarak kullanilabilir mi?

Garlic (Allium sativum L.) has been used in the public health in some of treatments for years. The most important chemical compounds of garlic are sulphide (alisin, ajoen and diallysulphure) compounds. Allisin is described as antibacterial, antifungal, antiparasitic and anticarsinogenic agent and many works practice on it. But its antiscolocidal effect has not been worked yet. In our work, whether garlic extract has effects both direct and daughter vesicules to protoscolex is searched. In the study during the surgery daughter vesicules and protoscolex which are acquired from liver cyst hydatique and extracts which obtained from garlic grown in the Kastamonu region are used. Viability determination is work at using 0.1% eozin solution. In 50% mg/ml concentration garlic extract to protoscolex' in 15 minutes, in 25% mg/ml concentration 20. minutes and 12.5% mg/ml concentration in 30 minutes have full effect. While it has full effect in 20. minutes to protoscolex into the vesicules within the 50% mg/ml 12.5 mg/ml concentration, there has been 80% viability in 40 minutes.

Antimicrobial activity of some endemic plants growing in the Eastern Black Sea Region, Turkey.

The Eastern Black Sea Region has an extensive flora because of ample rainfall lasting all year. In this study, antimicrobial effects of 74 crude extracts of 22 endemic plants were investigated. Among the 30 active crude extracts, water-insoluble crude extracts from Betula medwediewii, Heracleum platytaenium, Primula longipes, Anthemis cretica ssp. argaea and Centaurea helenioides were the prominent ones with their MIC values.

In vitro antioxidant activity of polyphenol extracts with antiviral properties from Geranium sanguineum L.

Recent evidence shows that plant polyphenols exhibit antioxidant and radical scavenging properties. By three separate and complementary methods--DPPH assay, beta-carotene-linoleic acid assay and NBT-reduction assay it was established that a polyphenol-rich extract from the medicinal plant Geranium sanguineum L. with strong anti-influenza virus activity, possessed antioxidant and radical scavenging capacities. For comparative reasons caffeic acid and the synthetic antioxidant BHT were used. Total soluble phenolic constituents of the MeOH extract measured by Folin-Ciocalteu reagent were found as 34.60% (w/w). Further it was demonstrated that the EtOAc fraction, retaining the majority of the in vivo protective effect exhibited a strong O2-scavenging activity while the n-BuOH fraction, containing the majority of the in vitro antiviral activity provoked generation of O2-. The O2- scavenging activity of all three preparations correlated with the rate of the protective effect shown in the murine model of experimental influenza virus infection. The present results are in accordance with our intensive studies on the mode of the protective effect of the plant extract which showed positively that the protection may possibly be attributed to the combination of more than one biological activities and that the use of antioxidants might be an useful approach in the treatment of influenza infection.

The in vitro antioxidant and antimicrobial activities of the essential oil and methanol extracts of Achillea biebersteini Afan. (Asteraceae).

The essential oil and methanol extracts from A. biebersteinii Afan. (Asteraceae) were evaluated for their antimicrobial and antioxidant activities in vitro. The oil showed stronger antimicrobial activity than the extracts. Their antioxidant features were also evaluated using diphenyl-1-picrylhydrazyl (DPPH), inhibition of superoxide and hydroxyl radicals and inhibition of the lipid peroxidation assays. Particularly, polar subfraction of the methanol extract showed antioxidant activity. The GC-MS analysis of the oil has resulted in the identification of 23 components; piperitone, eucalyptol, camphor, chrysanthenone and borneol were the main components. Antimicrobial activity tests carried out with the fractions of the oil showed that the activity was mainly observed in those containing eucalyptol and camphor, in particular, followed by borneol and piperitone.

In vitro antioxidant, antimicrobial, and antiviral activities of the essential oil and various extracts from herbal parts and callus cultures of Origanum acutidens.

The essential oil and various extracts obtained from Origanum acutidens and methanol extracts (MeOH) from callus cultures have been evaluated for their antioxidative, antimicrobial, and antiviral properties. The essential oil exhibited strong antimicrobial activity with a significant inhibitory effect against 27 (77%) of the 35 bacteria, 12 (67%) of the 18 fungi, and a yeast tested and moderate antioxidative capacity in DPPH and beta-carotene/linoleic acid assays. GC and GC-MS analyses of the oil resulted in the identification of 38 constituents, carvacrol being the main component. The MeOH extracts obtained from herbal parts showed better antioxidative effect than that of butylated hydroxytoluene (BHT), whereas callus cultures also exhibited interesting antioxidative patterns. Concerning antiviral activity, none of the extracts inhibited the reproduction of influenza A/Aichi virus in MDCK cells. The MeOH extracts from herbal parts inhibited the reproduction of HSV-1, and also callus cultures exerted slight antiherpetic effect.

In vitro antimicrobial and antioxidant activities of the essential oils and various extracts of Thymus eigii M. Zohary et P.H. Davis.

This study was designed to examine the in vitro antimicrobial and antioxidant activities of the essential oil and various extracts obtained from aerial parts of Thymus eigii. The essential oil was particularly found to possess stronger antimicrobial activity, whereas other nonpolar extracts and subfractions showed moderate activity and polar extracts remained almost inactive. GC-MS analysis of the oil resulted in the identification of 39 compounds, representing 93.7% of the oil; thymol (30.6%), carvacrol (26.1%), and p-cymene (13.0%) were the main components. The samples were also subjected to a screening for their possible antioxidant activity by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and beta-carotene-linoleic acid assays. In the former case, the polar subfraction of the methanol extract was found to be superior to all extracts tested, only 16.8 microg/mL of which provided 50% inhibition, whereas all extracts, particularly the polar ones, seem to inhibit the oxidation of linoleic acid in the latter case. These data were further supported by total phenolics analysis, indicating that the antioxidative potential of the extracts was closely related to their phenolic constituents.

Effects of Rhus coriaria L (Anacardiaceae) on lipid peroxidation and free radical scavenging activity.

In order to evaluate the antioxidant potential of Rhus coriaria L. (Anacardiaceae), a well-known spice, the methanolic extract (water-soluble part) was prepared and investigated using free radical-generating systems in vitro. The IC(50) value of the extract for lipid peroxidation was estimated as 1200 microg/mL in the Fe(+2)-ascorbate system while those for superoxide-scavenging activity in the xanthine-xanthine oxidase method and hydroxyl radical scavenging activity in the deoxyribose decomposition method were 282.92 microg/mL and 3850 microg/mL, respectively. These data suggest that the methanolic extracts of Rhus coriaria L. fruits have considerable antioxidant activity against free radicals and lipid peroxidation in vitro, a fact that may encourage in vivo studies.

Antimicrobial activity of essential oil and methanol extracts of Achillea sintenisii Hub. Mor. (Asteraceae).

The essential oil, obtained by Clevenger distillation, and water-soluble and water-insoluble parts of the methanol extracts of Achillea sintenisii Hub. Mor. were individually assayed for their antimicrobial activities against 12 bacteria and two yeasts, Candida albicans and C. krusei. No activity was exhibited by the water-soluble subfraction, whereas both the water-insoluble subfraction of the methanol extracts and the essential oil were found to be active against some test microorganisms studied. Since the essential oil possessed stronger activity than the other extracts tested, it was further fractionated and the fractions were evaluated for their antimicrobial activity, followed by GC-MS analysis, resulting in the identification of 32 compounds which constituted 90.2% of the total oil. The GC-MS analysis of the oil and its fractions revealed that the main components of the oil, e.g. camphor and eucalyptol, possessed appreciable activity against C. albicans and Clostridium perfringens. The fi ndings presented here also suggest that the other constituents of the oil, e.g. borneol and piperitone can also be taken into account for the activity observed.

Antioxidant and antimicrobial activity of the essential oil and methanol extracts of Achillea millefolium subsp. millefolium Afan. (Asteraceae).

The in vitro antimicrobial and antioxidant activities of the essential oil and methanol extracts of Achillea millefolium subsp. millefolium Afan. (Asteraceae) were investigated. GC-MS analysis of the essential oil resulted in the identification of 36 compounds constituting 90.8% of the total oil. Eucalyptol, camphor, alpha-terpineol, beta-pinene, and borneol were the principal components comprising 60.7% of the oil. The oil strongly reduced the diphenylpicrylhydrazyl radical (IC(50)=1.56 micro g/ml) and exhibited hydroxyl radical scavenging effect in the Fe(3+)-EDTA-H(2)O(2) deoxyribose system (IC(50)=2.7 micro g/ml). It also inhibited the nonenzymatic lipid peroxidation of rat liver homogenate (IC(50)=13.5 micro g/ml). The polar phase of the extract showed antioxidant activity. The oil showed antimicrobial activity against Streptococcus pneumoniae, Clostridium perfringens, Candida albicans, Mycobacterium smegmatis, Acinetobacter lwoffii and Candida krusei while water-insoluble parts of the methanolic extracts exhibited slight or no activity. This study confirms that the essential oil of Achillea millefolium possesses antioxidant and antimicrobial properties in vitro.

Antimicrobial and antioxidant activity of the essential oil and methanol extracts of Thymus pectinatus Fisch. et Mey. Var. pectinatus (Lamiaceae).

The essential oil, obtained by using a Clevenger distillation apparatus, and water-soluble (polar) and water-insoluble (nonpolar) subfractions of the methanol extract of Thymus pectinatus Fisch. et Mey. var. pectinatus were assayed for their antimicrobial and antioxidant properties. No (or slight) antimicrobial activity was observed when the subfractions were tested, whereas the essential oil showed strong antimicrobial activity against all microorganisms tested. Antioxidant activities of the polar subfraction and the essential oil were evaluated using 2,2-diphenyl-1-picrylhydrazyl, hydroxyl radical, superoxide radical scavenging, and lipid peroxidation assays. The essential oil, in particular, and the polar subfraction of the methanol extract showed antioxidant activity. The essential oil was analyzed by GC/MS, and 24 compounds, representing 99.6% of the essential oil, were identified: thymol, gamma-terpinene, p-cymene, carvacrol, and borneol were the main components. An antimicrobial activity test carried out with fractions of the essential oil showed that the activity was mainly observed in those fractions containing thymol, in particular, and carvacrol. The activity was, therefore, attributed to the presence of these compounds. Other constituents of the essential oil, such as borneol, gamma-terpinene, and p-cymene, could be also taken into account for their possible synergistic or antagonistic effects. On the other hand, thymol and carvacrol were individually found to possess weaker antioxidant activity than the crude oil itself, indicating that other constituents of the essential oil may contribute to the antioxidant activity observed. In conclusion, the results presented here show that T. pectinatus essential oil could be considered as a natural antimicrobial and antioxidant source.

Compositions and the in vitro antimicrobial activities of the essential oils of Achillea setacea and Achillea teretifolia (Compositae).

GC-MS analysis of the isolated essential oils from air-dried aerial parts of Achillea setacea and Achillea teretifolia, an endemic taxon, resulted in the identification of 51 constituents (79.8% of the total oil) and 42 constituents (87.1% of the total oil), respectively. Eucalyptol (1,8-cineole) was the major constituent of both oils studied (18.5 and 19.9%, respectively). The antimicrobial activities of the essential oils were individually evaluated against 14 microorganisms. Both oils exhibited inhibitory effects on Clostridium perfringens, Acinetobacter lwoffii and Candida albicans with a range of minimum inhibitory concentration values extended from 0.28 to 2.25 mg/ml. Camphor and their derivatives, borneol, terpinen-4-ol and eucalyptol (1,8-cineol) can be considered as the main antimicrobial constituents of the oils studied.