Chemical Composition and Antifungal and Antibiofilm Effects of Vitex pseudo-negundo Essential Oil against Pathogenic Fungal Strains.

Background: Vitex pseudo-negundo is a plant of the Lamiaceae family that grows in different parts of the world and the vicinity of seasonal rivers in Iran. Methods: The chemical composition of the Vitex pseudo-negundo essential oils was distilled and evaluated using gas chromatography/mass spectrometry. The antifungal activity of the essential oils against the fungal strains was analyzed by broth microdilution methods as suggested by the Clinical and Laboratory Standards Institute. Furthermore, the antibiofilm activity of the Vitex pseudo-negundo essential oils was assessed using the XTT reduction assay. Results: Based on GC/MS analysis, the major components of the Vitex pseudo-negundo essential oils were α-pinene, α-terpinyl acetate, limonene, and (E)-caryophyllene. The growth of tested yeasts was inhibited at concentrations ranging from 2 to 64 µl/mL. Vitex pseudo-negundo fruit essential oil was the most effective in inhibiting yeast growth. Moreover, the essential oils exhibited antifungal activity against filamentous fungi strains. Additionally, the biofilm formation of Candida albicans was inhibited by the leaf, flower, and fruit of the essential oils. Conclusion: Considering the significant antifungal activities of these essential oils, they can be considered a potential source for formulating novel agents to control fungal infections.

Evaluation of metabolites in Iranian Licorice accessions under salinity stress and Azotobacter sp. inoculation.

Licorice (Glycyrrhiza glabra L.) is an industrial medicinal plant that is potentially threatened by extinction. In this study, the effects of salinity (0 and 200 mM sodium chloride (NaCl)) and Azotobacter inoculation were evaluated on 16 licorice accessions. The results showed that salinity significantly reduced the fresh and dry biomass (FW and DW, respectively) of roots, compared to plants of the control group (a decrease of 15.92% and 17.26%, respectively). As a result of bacterial inoculation, the total sugar content of roots increased by 21.56% when salinity was applied, but increased by 14.01% without salinity. Salinity stress increased the content of glycyrrhizic acid (GA), phenols, and flavonoids in licorice roots by 104.6%, 117.2%, and 56.3%, respectively. Integrated bacterial inoculation and salt stress significantly increased the GA content in the accessions. Bajgah and Sepidan accessions had the highest GA contents (96.26 and 83.17 mg/g DW, respectively), while Eghlid accession had the lowest (41.98 mg/g DW). With the bacterial application, the maximum amounts of glabridin were obtained in Kashmar and Kermanshah accessions (2.04 and 1.98 mg/g DW, respectively). Bajgah and Kashmar accessions had higher amounts of rutin in their aerial parts (6.11 and 9.48 mg/g DW, respectively) when their roots were uninoculated. In conclusion, these results can assist in selecting promising licorice accessions for cultivation in harsh environments.

Therapeutic Efficacy of Great Plantain (Plantago major L.) in the Treatment of Second-Degree Burn Wounds: A Case-Control Study.

Background: Great plantain (Plantago major L. or P.major) is a medicinal plant that is available all around the world. The whole plant has several bioactive compounds including terpenoids, flavonoids, phenolic compounds, alkaloids, fatty acids, iridoid glycosides, polysaccharides, and vitamins. Scientific studies have recognized several medical benefits like wound healing, anti-inflammatory, antimicrobial, antiulcerative, and antioxidative agents. The wound-healing capacity of this plant has been investigated under in vivo and ex vivo conditions. In the current study, we aim to compare the therapeutic effect of the P.major extract with 1% sulfadiazine on the healing of second-degree burn wounds. Method: Second-degree burn victims were included in our study. The investigation and control group, respectively, received P. major ointment 10% and silver sulfadiazine ointment 1%. The bacterial culture from the wound site was taken on days 3, 7, 10, 13, and last day of hospitalization. Patients' subjective complaints were obtained through the visual analog scale (VAS). All patients were treated and evaluated in the hospital. Result: Among the 15 patients, 11 were male, and the mean age was 33.3 years. The average complete healing duration was 11.73 vs. 13 days in the P. major and control group, respectively (P=0.166). On the third day, infection control was similar between the two groups, and on the seventh day, all bacterial cultures were negative. Although there was a significant reduction in pain scores during the recovery time, no significant differences in pain reduction were noted between the two groups (P=0.849). Conclusion: We showed that P.major ointment is a safe and suitable herbal compound in the treatment of second-degree burn wounds that not only has wound-healing properties but also is an analgesic and antimicrobial compound.

Characterization and Physical and Biological Properties of Tissue Conditioner Incorporated with Carum copticum L.

AIM: One of the main problems in dentistry is the injury caused by the long-term application of an ill-fitting denture. The existence of multiple microorganisms along with the susceptibility of the tissue conditioners to colonize them can lead to denture stomatitis. This study is aimed at developing a tissue conditioner incorporated with Carum copticum L. (C. copticum L.) for the effective treatment of these injuries. MATERIALS AND METHODS: The Carum copticum L. essential oil composition was determined by gas chromatography-mass (GC-mass) spectrometry. The antimicrobial activity of the essential oil against the standard strains of bacterial and fungal species was determined by broth microdilution methods as suggested by the Clinical and Laboratory Standards Institute (CLSI). The physical and chemical properties of the prepared tissue conditioner were investigated by viscoelasticity, FTIR assays, and the release study performed. Furthermore, the antibiofilm activity of the Carum copticum L. essential oil-loaded tissue conditioner was evaluated by using the XTT reduction assay and scanning electron microscopy (SEM). RESULTS: The main component of the essential oil is thymol, which possesses high antimicrobial activity. The broth microdilution assay showed that the essential oil has broad activity as the minimum inhibitory concentration was in the range of 32-128 µg mL-1. The viscoelasticity test showed that the essential oil significantly diminished the viscoelastic modulus on the first day. The FTIR test showed that Carum copticum L. essential oil was preserved as an independent component in the tissue conditioner. The release study showed that the essential oil was released in 3 days following a sustained release and with an ultimate cumulative release of 81%. Finally, the Carum copticum L. essential oil exhibited significant activity in the inhibition of microbial biofilm formation in a dose-dependent manner. Indeed, the lowest and highest amounts of biofilm formation on the tissue conditioner disks are exhibited in the Streptococcus salivarius and Candida albicans by up to 22.4% and 71.4% at the 64 µg mL-1 concentration of C. copticum L. with a statistically significant difference (P < 0.05). CONCLUSION: The obtained results showed that the Carum copticum L. essential oil-loaded tissue conditioner possessed suitable physical, biological, and release properties for use as a novel treatment for denture stomatitis.

Development of pre-emergence herbicide based on Arabic gum-gelatin, apple pectin and savory essential oil nano-particles: A potential green alternative to metribuzin.

This study was conducted as a plot experiment to investigate the phytotoxicity effects of nano-encapsulated savory essential oil (EO) when it is incorporated separately into carbohydrate and protein natural polymers (Arabic gum-gelatin, apple pectin and gelatin) and two cross-linkers including one poly acid and one enzyme (citric acid and transglutaminase enzyme). Each product was tested as a pre-emergence herbicide against amaranth and tomato. The evaluations also involved determining the stability, morphology, encapsulation efficiency and release properties of the prepared formulations. Coating the savory EO with cross-linked biopolymers enhanced its stability and herbicidal activity, compared to the EO nano-emulsion without any polymer or cross-linker. Among the tested formulations, the strongest inhibitory effect against amaranth germination and growth was caused by Arabic gum-gelatin and apple pectin biopolymers at the concentration of 3 ml/L of EO, when cross-linked with citric acid. These two treatments had slight effects on tomato seedlings, however. The suppressive ability of the formulations was almost similar and comparable to the chemical herbicide metribuzin (1.75 g/L). In conclusion, Arabic gum-gelatin and apple pectin cross-linked by citric acid containing savory EO can be considered as potential, green and safe replacements for metribuzin in organic tomato production.

Study of two-stage ohmic hydro-extraction of essential oil from Artemisia aucheri Boiss.: Antioxidant and antimicrobial characteristics.

The effect of two-stage ohmic-assisted hydrodistillation (TSOH) on the extraction and characteristics of essential oils (EOs) from the Artemisia aucheri Boiss. was studied, and the results were compared to conventional hydrodistillation (HD). According to the results, the yield of EOs obtained through TSOH was almost 30% higher than those extracted by HD in nearly one-quarter of a time used by the HD. Scanning electron micrographs of A. aucheri leaves showed almost complete eruption of EO glands and their surrounding area in TSOH extraction method, hence achieving higher yield. The components of the EOs obtained through TSOH were only slightly different from those of HD. GC/MS analysis indicated some differences in the quantity of the main components, too. The main components of EOs were identified as Thymol, Linalool, Geraniol, Camphor, and 1, 8-Cineole, Davana ether and Cis-Davanone. Thymol (~17%) and Cis-Davanone (~23%) were the highest quantity in the EOs extracted from TSOH and HD, respectively. The variation of antioxidant and antimicrobial activities of the EOs may be attributed to these differences in the percentage of the main components. The radical scavenging activity of the EOs obtained by TSOH was almost twice that of HD. Based on antimicrobial activity assays, the EOs were efficient against S. aureus (a Gram-positive), E. coli (a Gram-negative), and S. cerevisiae (yeast). However, the efficacy was higher in gram-positive than gram-negative bacteria and yeast. The results indicate TSOH has a potential to produce EOs from herbal plants at a faster rate, higher yield, being probably more efficient in terms of energy although having similar antimicrobial and antioxidant efficacy.

Antimicrobial activity of seven essential oils from Iranian aromatic plants against common causes of oral infections.

BACKGROUND: Over the past two decades, there has been a growing trend in using oral hygienic products originating from natural resources such as essential oils (EOs) and plant extracts. Seven aromatic plants used in this study are among popular traditional Iranian medicinal plants with potential application in modern medicine as anti-oral infectious diseases. OBJECTIVES: This study was conducted to determine the chemical composition and antimicrobial activities of essential oils from seven medicinal plants against pathogens causing oral infections. MATERIALS AND METHODS: The chemical compositions of EOs distilled from seven plants were analyzed by gas chromatography/mass spectrometry (GC/MS). These plants included Satureja khuzestanica, S. bachtiarica, Ocimum sanctum, Artemisia sieberi, Zataria multiflora, Carum copticum and Oliveria decumbens. The antimicrobial activity of the essential oils was evaluated by broth micro-dilution in 96 well plates as recommended by the Clinical and Laboratory Standards Institute (CLSI) methods. RESULTS: The tested EOs inhibited the growth of examined oral pathogens at concentrations of 0.015-16 µL/mL. Among the examined oral pathogens, Enterococcus faecalis had the highest Minimum Inhibitory Concentrations (MICs) and Minimum Microbicidal Concentrations (MMCs). Of the examined EOs, S. khuzestanica, Z. multiflora and S. bachtiarica, showed the highest antimicrobial activities, respectively, while Artemisia sieberi exhibited the lowest antimicrobial activity. CONCLUSIONS: The excellent antimicrobial activities of the tested EOs might be due to their major phenolic or alcoholic monoterpenes with known antimicrobial activities. Hence, these EOs can be possibly used as an antimicrobial agent in treatment and control of oral pathogens.

In vivo study of the efficacy of the aromatic water of Zataria multiflora on hydatid cysts.

Gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) were employed to determine the chemical composition of the essential oil (EO) from aromatic water (AW) of Zataria multiflora. Thymol (66.9%), carvacrol (15.2%), and carvone (7.3%) were found to be the major EO constituents. Eighty laboratory BALB/c mice were infected intraperitoneally by injection of 1,500 viable protoscolices and were divided into prevention (40 mice) and therapeutic (40 mice) groups. To prove the preventive effect of the Z. multiflora AW on development of hydatid cysts, the 40 infected mice were allocated into three treatment groups, including the albendazole group (10 mice that received 150 mg/kg body weight/day for 10 days), the Z. multiflora AW group (15 mice that received 20 ml/liter in drinking water for 8 months), and a control group (15 mice that received no treatment). To estimate the therapeutic effect of the Z. multiflora AW on the hydatid cyst, after 8 months of infection, the 15 remaining mice were allocated into three experimental treatment groups of five animals each, including the albendazole group (300 mg/kg/day for 20 days), Z. multiflora AW group (40 ml/liter in drinking water for 30 days), and control group (no treatment). All mice were then euthanized, and the sizes and weights of the cysts as well as their ultrastructural changes were investigated. The weights and sizes of the hydatid cysts significantly decreased upon treatment with the Z. multiflora AW in both the preventive and therapeutic groups (P < 0.05). The results of scanning electron microscopy also showed considerable damage in the germinal layer of the hydatid cysts recovered from the treated animals.

Preventive and therapeutic effects of Zataria multiflora methanolic extract on hydatid cyst: an in vivo study.

The phenolic compounds of Zataria multiflora extract, were identified by HPLC analysis. Gallic acid, catechin, caffeic acid, and quercetin were found to be the major phenolic compounds. Eighty healthy laboratory Balb/C mice were infected intraperitoneally by injection of 1500 viable protoscoleces and were divided into prevention (40 mice) and therapeutic (40 mice) groups. To prove the preventive effect of Z. multiflora extract on development of hydatid cyst, the 40 infected animals were allocated into three treatment groups including Z. multiflora (4 g/l in drinking water for 8 months), albendazole (150 mg/kg BW/day for 10 days) and untreated (control) group. To estimate the therapeutic effect of Z. multiflora extract on the hydatid cyst, after 8 months of infection, the infected mice were allocated into three experimental treatment groups including Z. multiflora (8 g/l in drinking water for 30 days), albendazole (300 mg/kg BW/day for 20 days) and untreated (control) group. At the end of the treatment period, all mice were euthanized and necropsied, the hydatid cysts were carefully removed, weighed and their size were recorded. Weight and size of the hydatid cysts significantly decreased (p<0.05) upon the treatment with Z. multiflora extract in both prevention and therapeutic groups. The germinal layer of the hydatid cysts recovered from the treated mice, either from the prevention or therapeutic group, were completely damaged at ultrastructural level by scanning electron microscopy.

Chemical composition and antimicrobial activities of essential oil of nepeta cataria L. Against common causes of oral infections.

OBJECTIVES: Over the past two decades, there has been a growing trend in using oral hygienic products from natural resources such as essential oils and plant extracts. Nepeta cataria L. is a member of the mint family (Labiatae) with several medicinal properties. The objective of this study was to determine the chemical composition and antimicrobial activities of essential oils (EOs) from N. cataria leaves against pathogens causing oral infections. MATERIALS AND METHODS: The chemical composition of EOs from N. cataria was analyzed by gas chromatography/mass spectrometry (GC/MS). The antimicrobial activity of the essential oil was evaluated by broth micro-dilution in 96 well plates as recommended by the Clinical and Laboratory Standards Institute (CLSI) methods. The plates were incubated at 30°C for 24-48 h (fungi) or at 37°C for 24 h (bacteria). RESULTS: The analysis of the EOs indicated that 4a-α, 7-α, 7a-ß-nepetalactone (55-58%), and 4a-α, 7-ß, 7a-α-nepetalactone (30-31.2%) were the major compounds of the EOs at all developmental stages. The tested EOs exhibited antimicrobial activities against the tested bacteria at concentrations of 0.125-4 µL/mL. Moreover, the oils entirely inhibited the growth of Candida species at a concentration less than 1 µL/mL. CONCLUSION: Based on these results, the EO of N. cataria can possibly be used as an antimicrobial agent in the treatment and control of oral pathogens.

In vitro lethal effect of ajowan (Trachyspermum ammi L.) essential oil on hydatid cyst protoscoleces.

Various chemical scolicidal agents have been used for inactivation of hydatid cyst protoscolices, but most of them are associated with adverse side effects. Since ajowan (Trachyspermum ammi) has been shown to have a number of medicinal properties, in this study the scolicidal effect of the essential oil (EO) from the fruits of this herbal plant was investigated. Ajowan EO was obtained by hydrodistillation method. Gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) were employed to determine the chemical composition of the EO. Protoscoleces were exposed to various concentrations of EO (3, 5 and 10mg/mL) for 10, 20, 30, and 60 min. Viability of protoscolices was confirmed by 0.1% eosin staining. A total of 18 compounds representing 99.54% of the total oil, were identified. Thymol (50.07%), γ-terpinene (23.92%), and p-cymene (22.9%) were found to be the major EO constituents. While the mortality rate of protoscolices in the control group was 6.67%, scolicidal power of ajowan EO at concentration of 3mg/mL was 31.34, 35.98, 45.17, and 51.58% after 10, 20, 30, and 60 min, respectively. The EO at concentration of 5mg/mL killed 51.89, 72.20, 88.64, and 100% of protoscolices after 10, 20, 30, and 60 min, respectively. One hundred percent scolicidal activity was observed with ajowan EO at concentration of 10mg/mL after 10 min of exposure. The results of this study revealed that the EO of ajowan is rich in thymol, γ-terpinene and p-cymene, has high scolicidal power and it may be used as a natural scolicidal agent.

Effect of Satureja khuzestanica essential oil on oxidative stability of sunflower oil during accelerated storage.

In this study, the application of various concentrations (0.02%, 0.04%, 0.06% and 0.08%) of Satureja khuzestanica essential oil (EO) was examined on the oxidative stability of sunflower oil and compared to butylated hydroxyanisole (BHA) during storage at 60°C. Gas chromatography (GC) and GC-mass spectrometry analyses of the oils revealed that carvacrol (87.7%) was the major component of EO. Peroxide value and anisidine value measurements in sunflower oil showed that all concentrations of EO had antioxidant effects in comparison to BHA. Oil samples supplemented with EO concentration of 0.08% were the most stable during storage (p < 0.05). EO also was able to reduce the stable free radical 2, 2-diphenyl-1-picrylhydrazyl with a 50% inhibition concentration (IC50) of 31.5 ± 0.6 µg mL⁻¹. Therefore, the results indicate that EO could be used as a natural antioxidant in food lipids.

Influence of growth phase on the essential oil composition and antimicrobial activities of Satureja hortensis.

The variations in quantity and quality of essential oils (EOs) from the aerial parts of cultivated Satureja hortensis were determined at different stages of harvesting. The EOs of air-dried samples were obtained by hydrodistillation and analyzed by gas chromatography/mass spectrometry (GC/MS). The antimicrobial activity of the EOs was investigated by broth microdilution methods. The amount of EOs (w/w, %) were 2.3, 2.5, 2.0, and 1.8% at floral budding, full flowering, immature fruit, and ripened fruit stages, respectively. gamma-Terpinene was the major compound of the EO at all developmental stages, except the ripened fruit stage when it was replaced by carvacrol (46.4%). The EOs exhibited strong antibacterial activities against the tested bacteria. Moreover, the EOs either inhibited or killed the examined yeasts at concentrations ranging from 0.03-8.0 microL/mL. Considering the wide range of antimicrobial activities of the examined EOs, they might have potential to be used in the management of infective agents.