Exposure to single-walled carbon nanotubes differentially affect in vitro germination, biochemical and antioxidant properties of Thymus daenensis celak. seedlings.

Carbon nanomaterials such as single-walled carbon nanotubes (SWCNTs) offer a new possibility for phyto-nanotechnology and biotechnology to improve the quality and quantity of secondary metabolites in vitro. The current study aimed to determine the SWCNTs effects on Thyme (Thymus daenensis celak.) seed germination. The seedlings were further assessed in terms of morphological and phytochemical properties. Sterile seeds were cultured in vitro and treated with various concentrations of SWCNTs. Biochemical analyses were designed on seedling sample extracts for measuring antioxidant activities (AA), total flavonoids (TFC) and phenolic contents, and the main enzymes involved in oxidative reactions under experimental treatments. The results indicated that an increase in SWCNTs concentration can enhance the total percentage of seed germination. The improvement was observed in samples that received SWCNTs levels of up to 125 µg ml-1, even though seedling height and biomass accumulation decreased. Seedling growth parameters in the control samples were higher than those of grown in SWCNT-fortified media. This may have happened because of more oxidative damage as well as a rise in POD and PPO activities in tissues. Additionally, secondary metabolites and relevant enzyme activities showed that maximum amounts of TPC, TFC, AA and the highest PAL enzyme activity were detected in samples exposed to 62.5 µg ml-1 SWCNTs. Our findings reveal that SWCNTs in a concentration-dependent manner has different effects on T. daenensis morphological and phytochemical properties. Microscopic images analysis revealed that SWCNTs pierce cell walls, enter the plant cells and agglomerate in the cellular cytoplasm and cell walls. The findings provide insights into the regulatory mechanisms of SWCNTs on T. daenensis growth, germination and secondary metabolites production.

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.

The use of silicon and mycorrhizal fungi to mitigate changes in licorice leaf micromorphology, chlorophyll fluorescence, and rutin content under water-deficit conditions.

In this study, the effects of water-deficit conditions, silicon (Si) fertilizer (300 ppm), and arbuscular mycorrhizal (AM) inoculation by Claroiedoglomus etunicatum were evaluated on several features of licorice (Glycyrrhiza glabra L.). The measurable features were photosynthetic parameters, rutin content in aerial parts, and leaf micromorphology. Drought was administered at five levels determined by the percentage of field capacity (FC), i.e. 100, 80, 60, 40, and 20% of FC. Leaf extracts were utilized for measuring rutin content (via HPLC), and photosynthetic pigments; measurement of stomatal density, and trichome analysis were performed by scanning electron microscopy (SEM). Under severe drought stress, leaf area decreased by 50.84%, compared to well-irrigated plants. A significant decrease in leaf numbers (32.52%) was observed because of deficit irrigation. AM and Si improved chlorophyll fluorescence, which corresponded to the maximum efficiency of photosystem II. Rutin content decreased significantly under deficit irrigation. Also, the integration of AM and Si treatments positively affected rutin quantity under various irrigation regimes. Under moderate stress (60% FC), using AM and/or Si treatments reduced the stomatal length by 61.22 and 52.98%, respectively. Interestingly, a significant reduction in stomatal density towards control was observed as a result of the integrated treatments of Si and AM (58.28% at W20 and 59.82% at W100), which helped plants reduce water loss when facing drought stress. Principal component analysis (PCA) showed that photosynthetic pigments, chlorophyll fluorescence, and rutin changed quantitatively under moderate drought stress, while more variations were observed in leaf epidermal micromorphology under severe drought stress. These findings revealed that Si and AM, by exogenous application, synergistically mitigated the effects of drought stress on licorice.

Adaptation of Glycyrrhiza glabra L. to water deficiency based on carbohydrate and fatty acid quantity and quality.

Water deficit affects agricultural systems negatively globally. This research objective was to mitigate drought's detrimental effects on plants metabolite profiling by utilizing biofertilizers and mineral nutrition. The carbohydrate content and fatty acid profile of Licorice (Glycyrrhiza glabra) were assessed under Silicon (Si) nutrition, Claroiedoglomus etunicatum inoculation (F), and drought stress (100, 80, 60, 40, and 20% of field capacity (FC)). Results showed that Si application increased total sugar content under severe drought levels (20 and 40% FC) and made it reach 12.41 and 12.63 g/100 g DW, respectively. Sucrose, as the predominant sugar of licorice, was at its highest level (13.1 g/100 g DW) in response to integrated values of F and Si (60% FC). Gas chromatography-mass spectrometry showed that the majority of fatty acid components in plants were 9-Octadecenoic acid (8.72-71.27%), 9,12-Octadecadienoic acid (0.1-56.43%), Hexadecanoic acid (12.84-30.59%), Octadecanoic acid (6.9-15.3%), Docosanoic acid (0.57-2.77%), Eicosanoic acid (1.07-2.64%), and 7-Hexadecenoic acid (0.26-2.62%). Since a lower omega6/omega3 ratio represents a healthier product, the lowest ratio (0.25%) was observed in well-watered inoculated plants. Also, severe drought-treated plants under integrated Si and F applications showed a low omega6/omega3 ratio (1.88%). In conclusion, Si and F improved synergistically the carbohydrate content and fatty acid profile in plants, despite the drought stress.

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.

Microbial amelioration of salinity stress in endangered accessions of Iranian licorice (Glycyrrhiza glabra L.).

BACKGROUND: Glycyrrhiza glabra L. is a medicinal and industrial plant that has gone extinct due to different abiotic stress caused by climate change. To understand how the plant-associated microorganism can support this plant under salinity, we collected sixteen Iranian accessions of G. glabra L., inoculated their rhizomes with Azotobacter sp. (two levels, bacterial treatments, and no-bacterial treatments, and grown them under salinity stress (NaCl levels; 0, and 200 mM). RESULTS: Two accessions of Bardsir and Bajgah significantly showed higher resistant to salinity, for example by increasing crown diameter (11.05 and 11 cm, respectively) compared to an average diameter of 9.5 in other accessions. Azotobacter inoculation caused a significant increase in plant height and crown diameter. Among studied accessions, Kashmar (46.21%) and Ilam (44.95%) had the highest rate of membrane stability index (MSI). Evaluation of enzyme activity represented that bacterial application under salinity, increased polyphenol oxidase (PPO) (0.21 U mg-1 protein), peroxidase (POD) (3.09 U mg-1 protein U mg-1 protein), and phenylalanine ammonia-lyase (PAL) (17.85 U mg-1 protein) activity. Darab accession showed the highest increase (6.45%) in antioxidant potential compared with all studied accessions under Azotobacter inoculation. According to principal component analysis (PCA), it was found that the accession of Meshkinshahr showed a more remarkable ability to activate its enzymatic defense system under salt stress. Also, three accessions of Meshkinshahr, Eghlid, and Ilam were categorized in separated clusters than other accessions regarding various studied treatments. CONCLUSION: Analysis indicated that five accessions of Meshkinshahr, Rabt, Piranshahr, Bardsir, and Kermanshah from the perspective of induced systematic resistance are the accessions that showed a greater morphophysiological and biochemical outcome under salinity. This study suggested that, inoculation of with Azotobacter on selected accession can relieve salt stress and support industrial mass production under abiotic condition.

Eco-friendly 'ochratoxin A' control in stored licorice roots - quality assurance perspective.

According to toxicity data, ochratoxin A (OTA) is the second most important mycotoxin and is produced by Aspergillus and Penicillium. As a natural antifungal agent, clove essential oil (CEO) is a substance generally recognised as safe (GRAS) and shows strong activity against fungal pathogens. Here, we aimed to investigate the control efficacy of CEO in nano-emulsions (CEN) against OTA production in licorice roots and rhizomes during storage. The experiments were performed under simulated conditions of all four seasons (i.e. Spring, Summer, Autumn and Winter). Relative humidity (RH) and temperature were simulated in desiccators along with various salt solutions in incubators. Fresh licorice roots were immersed in CEN at various concentrations (150, 300, 600, 1200 and 2400 µl/l). Before utilising the nano-emulsions, we measured their polydispersity index and mean droplet size by the dynamic light scattering (DLS) technique. Also, the chemical composition of the CEO was determined using GC and GC-MS analyses. Sampling was carried out to monitor OTA once every five days. The samples were dried immediately and analysed by high-performance liquid chromatography (HPLC). Results showed that various concentrations of CEN inhibited the growth of fungi and OTA production. The most effective CEN concentrations were 1200 and 2400 µl/l, which reduced OTA production to 19 and 20 ppb under Winter and Autumn conditions, respectively. These results suggest an effective eco-friendly method for the storage of licorice to reduce postharvest fungal decay.

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.

Antimicrobial core-shell electrospun nanofibers containing Ajwain essential oil for accelerating infected wound healing.

Treatment of skin injuries is still facing major challenges, such as chronicity and infections, particularly those caused by multi-drug resistance pathogens. An effective treatment of such wounds should accelerate the wound healing process while preventing bacterial contamination. Here, a novel core-shell nanofiber mat was fabricated comprising gelatin/polyvinyl alcohol (as a core) and aloe vera/arabinose/polyvinylpyrrolidone (as a shell) for accelerating the healing process of bacteria-infected wounds. Trachyspermum Ammi (Ajwain) essential oil (EO), as a potent and natural antimicrobial agent against microorganisms, was incorporated into the core of nanofiber mats using coaxial electrospinning. The microscopy images demonstrated the successful fabrication of the core-shell structure with a uniform fiber size of 564 ± 106.35 nm. Moreover, Ajwain EO-loaded nanofiber mat (core-shell/EO) provided excellent antimicrobial activity and antioxidant ability. The in vitro and ex vivo release of Ajwain EO from the fabricated nanofiber mat corroborated a prolonged release profile. Furthermore, in vivo antibacterial activity, wound closure, and histomorphological examinations showed the high efficacy of the core-shell/EO mat in the treatment of Staphylococcus aureus-infected full-thickness rat wounds compared to standard control treatment with a gauze. Overall, these results represent the core-shell/EO mat's potential as a newly developed wound dressing for bacteria-infected full-thickness skin injuries.

Core-shell chitosan/PVA-based nanofibrous scaffolds loaded with Satureja mutica or Oliveria decumbens essential oils as enhanced antimicrobial wound dressing.

Wounds are prone to bacterial infections, which cause a delayed healing process. Regarding the emergence of bacterial resistance to common antibiotics, using natural antimicrobial agents can be beneficial. Chitosan is a biological polymer, which has shown partial antioxidant and antimicrobial activities. In this study, core-shell nanofibrous scaffolds composed of chitosan (CS)/polyvinyl alcohol (PVA) as the core and polyvinylpyrrolidone (PVP)/ maltodextrin (MD) as the shell were developed. Satureja mutica (S. mutica) or Oliveria decumbens (O. decumbens) essential oil (EO) was encapsulated into the core of the produced scaffolds. The broth microdilution analysis showed significant antimicrobial activity of the EOs. The SEM analysis indicated that the unloaded and loaded core-shell scaffolds with S. mutica or O. decumbens EO had a uniform, beadless structure with fiber mean diameters of 210 ± 50, 250 ± 45, and 225 ± 46 nm, respectively. The CS/PVA-PVP/MD and CS/PVA/EO-PVP/MD scaffolds indicated suitable mechanical properties. The addition of the studied EOs enhanced the antioxidant activity of the scaffolds. The antimicrobial test of produced scaffolds showed that loading of 10% S. mutica or O. decumbens EO could broaden the microbicidal activity of the CS/PVA-PVP/MD scaffolds. These results revealed that the CS/PVA/EO-PVP/MD nanofibrous scaffolds are promising candidates for wound dressing.

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.

A natural post-emergence herbicide based on essential oil encapsulation by cross-linked biopolymers: characterization and herbicidal activity.

This work describes efforts to encapsulate savory (Satureja hortensis L.) essential oil (EO) with different natural polymers (i.e., Arabic gum/gelatin (AGG), apple pectin (AP), gelatin (G)) and, as a separate set of experiments, with bio cross-linkers (i.e., citric acid and transglutaminase enzyme). The phytotoxic activity of encapsulated savory EO on tomato (Lycopersicon esculentum Mill.) and amaranth weed (Amaranthus retroflexus L.) was investigated. The micro-capsules were evaluated in terms of size, polydispersity, stability, encapsulation efficiency, morphology, and release properties. The Korsmeyer-Peppas model operated when EO was being released from the micro-capsules. Carvacrol (52.5%) and γ-terpinene (30.2%) comprised the main constituents of the savory EO. Based on the results, encapsulating the EO with cross-linked biopolymers increased the stability and herbicidal activity of EO, as compared to simple EO emulsions. Maximum toxicity injuries (MTI) were caused by encapsulations of apple pectin, cross-linked with APe enzyme (15 ml/L) on both plant species. MTI were observed 2 days after using the micro-encapsulated herbicides (MCHs). However, the injury caused by MCHs on tomato was not significant. The lowest values of fresh weight (2.80 g), chlorophyll a (0.194 mg/g Fw), and total chlorophyll content (0.219 mg/g Fw) of amaranth occurred in response to APe (15 ml/L). Moreover, using AP(e) (10 ml/L) caused the lowest values of starch (0.444 mg/g Fw) and flavonoid contents (4.18 mg Cat/g Fw) in amaranth which measured as 59% and 90% reductions, respectively, in comparison with the control. The highest values of MDA (0.0109 nmol/g Fw) and H2O2 (0.0432 µmol/g Fw) were observed in amaranth plants treated with AP(e) (10 ml/L). In summary, cross-linked apple pectin can perform well in slow release delivery systems of agrochemicals. It can be recommended for use in the production of commercial, EO-based natural herbicides.

Multi-walled carbon nanotubes stimulate growth, redox reactions and biosynthesis of antioxidant metabolites in Thymus daenensis celak. in vitro.

This research was aimed at determining the effects of multi-walled carbon nanotubes (MWCNTs) on seed germination, seedling growth parameters and secondary metabolite (SM) production of Thymus daenensis in vitro. Seeds were aseptically cultured in Murashige and Skoog medium (MS) with various concentrations of MWCNTs (0, 125, 250, 500, 1000 and 2000 µg ml-1). Seed germination and morphological changes in seedlings were measured. The measurements were aimed at quantifying the total phenolic contents (TPC) and flavonoids (TFC), antioxidant activities and the activity of polyphenol oxidase (PPO), l-phenylalanine ammonia-lyase (PAL), dehydrogenase (DHA) and peroxidase enzyme (POD) of the seedling extract. Seedling biomass and seedling height grew significantly as the MWCNTs level increased. The biomass and height peaked at 250 µg ml-1 (0.41 ± 0.01 gr FW, 5.99 ± 0.55 cm) and then rapidly decreased to 0.040 ± 0.1 gr FW and 1.42 ± 0.24 cm in response to 1000 µg ml-1, 30 days after the treatment. Additionally, SM and the analyses of enzyme activity revealed that the highest amounts of TPC (6.70 ± 0.06 mg GAE g-1 DW), TFC (8.19 ± 0.01 mg QUE g-1 DW), antioxidant activities (73.88 ± 0.47%) and maximum PAL activity (1.25 ± 0.08 mM cm g-1 FW) were detected in plants grown on MS media fortified with 250 µg ml-1 MWCNTs. The results reveal that MWCNTs in low doses (250 µg ml-1) can encourage the production of biomass, elicit more SM from seedlings and enhance the biosynthesis of antioxidants. TEM images showed that MWCNTs could cross the plant cell wall and enter the cellular cytoplasm.

The Lethal Effect of a Nano Emulsion of Satureja hortensis Essential Oil on Protoscoleces and Germinal Layer of Hydatid Cysts.

BACKGROUND: New scolicidal agents and novel therapeutic drugs are essential for better management of the zoonotic infection, hydatid disease. This study evaluated the effect of a nanoemulsion (NE) of Satureja hortensis essential oil (SHEO) on protoscoleces and germinal layer of hydatid cysts. METHODS: This study was conducted from July to October 2016 in Shiraz University, Shiraz, Iran. Gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) were performed to identify the main components of SHEO. To determine the scolicidal power of the NE of SHEO, live protoscoleces of hydatid cysts were exposed to two concentrations (1 and, 2 mg/mL) of the NE and incubated at 37 °C for 10 and 20 min. To evaluate the anti-hydatid effect of the NE of SHEO, the collected hydatid cysts from the abdominal cavities of the experimentally infected mice were immersed in the NE (0.5 mg/ml) and incubated at 22 °C for 24 h. RESULTS: Carvacrol and γ-terpinene were the major components of the SHEO. NE of SHEO at the concentrations of 1 and 2 mg/mL showed 100% scolicidal power after 20 and 10 min respectively. Exposure of the hydatid cysts to the NE of SHEO resulted in crumpling of their germinal layer and detachment of this layer from the laminated layer. CONCLUSION: NE of SHEO showed a strong scolicidal activity as well as a profound lethal effect on the germinal layer of hydatid cysts. Accordingly, this product may be used as a natural scolicidal agent in hydatid cyst surgery. Furthermore, it may be used as a therapeutic tool for treatment of hydatid disease.

Chemical compositions and antifungal activities of Satureja macrosiphon against Candida and Aspergillus species.

BACKGROUND AND PURPOSE: Despite the various applications of Satureja species, there are limited data in this domain. Regarding this, the present study was conducted to investigate the essential oil (EO) biological activity of S. macrosiphon species in Iran. MATERIALS AND METHODS: The EO of S. macrosiphon flowers was obtained by hydrodistillation. Chemical compositions of the EO were analyzed using gas chromatography-mass spectrometry. In addition, minimum inhibitory concentrations (MIC) were measured by means of the broth microdilution method. The estimation of antibiofilm and cytotoxic activities was also accomplished using the tetrazolium salt and MTT assays, respectively. RESULTS: A total of 26 components were identified in the EO with linalool as the main constituent (28.46%). A MIC range value of 0.25-8 µL/mL was obtained against all of the tested fungi. The EO inhibited the biofilm development of the Candida tested strains at a concentration of 4-8 µL/mL. Cytotoxicity (IC50) of EO against the HeLa cell was greater than the MIC concentration (6.49 µL/mL). CONCLUSION: Based on the findings, it was concluded that the EO of S. macrosiphon has the potential for further use as an antifungal agent.

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.

In vitro ovicidal activity of Peganum harmala seeds extract on the eggs of Fasciola hepatica.

Peganum harmala seeds extract has been previously reported to have antimicrobial and other medicinal properties. The aim of this study was to evaluate the ovicidal activity of the methanolic extract of P. harmala seeds against the eggs of F. hepatica. The phenolic compounds of the methanolic extract of P. harmala seeds were identified by HPLC analysis. Catechin, rutin, p-Coumaric acid, chloregenic acid and hesperetin were found to be the major phenolic compounds. F. hepatica eggs were collected from the gall bladder of naturally infected sheep. The eggs were exposed to two concentrations of P. harmala seeds extract (1 and 3 mg/mL) for 24 and 48 h. To investigate the effect of the P. harmala seeds extract on the miracidial formation, the treated eggs were incubated at 28 °C for 14 days. The results indicated that F. hepatica eggs were susceptible to the methanolic extract of P. harmala seeds. Following 24 h exposure of the eggs to P. harmala seeds extract with concentrations of 1 and 3 mg/mL, the miracidial formation reduced to 5 and 2.2 % respectively (compared with 60 % for the control group). Following 48 h of exposure of the eggs to P. harmala seeds extract with 1 mg/mL concentration, the miracidial formation reduced to 0.5 %. In this exposure time, no miracidial formation was observed in the eggs exposed to P. harmala seeds extract with concentration of 3 mg/mL. Therefore, the results of this study indicated that P. harmala seeds extract has high ovicidal activity against the eggs of F. hepatica. Accordingly, this extract may have the potential flukicidal activity against the immature and mature F. hepatica.

In vitro and in vivo antihydatid activity of a nano emulsion of Zataria multiflora essential oil.

In in vitro process of this study, protoscoleces of the hydatid cysts were exposed to two concentrations of nano emulsion (NE) of Zataria multiflora essential oil (ZMEO) (1 and 2mg/mL) for 10 and 20min. Viability of protoscoleces was confirmed using 0.1% eosin staining. For in vivo studies, sixteen laboratory mice were infected intraperitoneally by 1500 live protoscoleces. Five months after infection, the infected mice were divided into treatment and control groups. The mice of treatment group received the NE of ZMEO (20mg/kg) orally via their drinking water while the mice of control group received no treatment. Two months after the start of treatment, all of the mice were necropsied and the hydatid cysts were collected. Subsequently, the numbers, sizes and weights of the collected cysts were compared between the mice of two groups. The results of in vitro scolicidal assays showed that the scolicidal power of NE of a ZMEO at concentration of 1mg/mL was 88.01%, and 100% after 10 and 20min respectively. NE of ZMEO showed 100% scolicidal power at a concentration of 2mg/mL after 10min (comparing to 4.46% for the control group). The results of in vivo studies revealed that the size of the largest cysts as well as the total number of the cysts were significantly lower in the mice treated with NE of ZMEO (P<0.05). In conclusion, NE of ZMEO may be considered as a natural scolicidal agent and a potential therapeutic tool for treatment of hydatid disease.

Natural herbicide activity of Satureja hortensis L. essential oil nanoemulsion on the seed germination and morphophysiological features of two important weed species.

The aim of the present study was to obtain an oil/water (O/W) nanoemulsion (NE) containing garden savory (Satureja hortensis) essential oil (EO) and evaluating its herbicidal activity against Amaranthus retroflexus and Chenopodium album. Gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) were employed to determine the chemical composition of the EO. Carvacrol (55.6%) and γ-terpinene (31.9%) were the major EO components. Low energy method was applied, allowing achievement of EO nanodroplets. The NE also presented low polydispersity, and the mean droplet was below 130nm even after storage for 30d. Laboratory tests showed that the NE at different concentrations (100, 200, 400, 800, and 1000µL.L-1) significantly (P≤0.05) reduced the germination indices and the seedling's growth in dose-response. The inhibitory effect was the greatest at 800µL.L-1 NE. Overall, root length was more inhibited as compared to shoot length. Post-emergence application of NE at different concentrations (1000, 2000, 3000, 4000 and 5000µL.L-1 of EO) on 2-4 true leaves' stage of the weeds caused significant (P≤0.05) decrease in the growth factors in dose-dependent manner. Complete lethality was observed by 4000µL.L-1 NE sprayed on the weeds. Spraying of NE significantly (P≤0.05) reduced chlorophyll content in the tested weeds. Increasing in relative electrolyte leakage (REL) 1 and 5d after treatment represented significant cell membrane disruption and increased cell membrane permeability. Transmission electron microscope (TEM) pictures confirmed NE droplet size and demonstrated membrane destruction. The study approved that the NE of S. hortensis EO has herbicidal properties as it has high phytotoxic effect, and interferes with the germination, growth and physiological processes of the weeds. The production of NE from S. hortensis EO is a low energy method that offers a promising practical natural herbicide for weed control in organic agricultural systems.