Transcriptome analysis of Reticulitermes flaviceps exposed to Mentha spicata essential oil and carvone indicates a potential neurotoxic mechanism of action characterized by intracellular calcium influx mediated by dopamine receptors.

Reticulitermes flaviceps is an economically important pest in agriculture, forestry, and construction. Recent studies have shown an increase in research focusing on the anti-termite properties of plant essential oils, however, there remains a lack of information regarding the specific molecular mechanism involved. In this study, RNA-seq analysis was conducted on termites exposed to Mentha spicata essential oil (EO) and carvone, leading to the discovery of various genes that were expressed differentially under different treatment conditions. Numerous genes that exhibited a response to M. spicata EO and carvone found to be associated with stress-related pathways, such as drug metabolism cytochrome P450, glutathione metabolism, fatty acid metabolism, citric acid cycle, neuroactive ligand-receptor interaction, cell apoptosis, the AMPK signalling pathway, the mTOR signalling pathway, the longevity regulation pathway, ubiquitin-mediated protein hydrolysis, and the calcium signalling pathway. The up-regulation of genes (SPHK) associated with calcium channels, such as SPHK, indicates a potential mechanism of neurotoxicity, while the up-regulation of apoptosis-associated genes, including ACTB_G1, PYG, SQSTM1, RNF31, suggests a potential mechanism of cytotoxicity. The metabolism of M. spicata EO induces oxidative stress, elevates free Ca2+ levels in mitochondria, and initiates the generation of reactive oxygen species (ROS), ultimately resulting in programmed cell necrosis and apoptosis, as well as facilitating cellular autophagy. The monoterpenes exhibited neurotoxic and cytotoxic effects on R. flaviceps and could be exploited to advance termiticide development and eco-friendly termite control.

Systematic metabolic engineering for improved synthesis of perillic acid in Candida tropicalis.

Perillic acid has been studied as an anticancer and antimicrobial drug. Production of perillic acid has attracted considerable attention. Meanwhile, Candida tropicalis is an unconventional diploid yeast, most significantly characterized by its ability to metabolize alkanes or fatty acids for growth and proliferation. Therefore, perillic acid's precursor (L-limonene) in C. tropicalis was firstly synthesized by expressing a Mentha spicata L-limonene synthase gene, LS_Ms in this work. Expression of a gene which encoded for a truncated version of tLS_Ms increased the production of L-limonene with a 2.78-fold increase in the titer over C. tropicalis GJR-LS-01. Compartmentalized expression of the gene tLS_Ms inhibited the production of L-limonene in C. tropicalis compared to cytoplasmic expression. Cytoplasmic overexpression of seven precursor synthesis genes significantly enhanced the production of L-limonene in C. tropicalis compared to their compartmentalized expression (mitochondria or peroxisomes), which increased by 31.7-fold in C. tropicalis GJR-tLS-01. The L-limonene titer in C. tropicalis GJR-EW-tLS-04 overexpressing the mutant gene ERG20WW in the cytoplasm was significantly increased, 11.33-fold higher than the control. The titer of L-limonene for 60 g/L glucose was increased by 1.40-fold compared to the control. Finally, a Salvia miltiorrhiza cytochrome P450 enzyme gene CYP7176 and an Arabidopsis thaliana NADPH cytochrome P450 reductase gene CPR were heterologously expressed in C. tropicalis GJR-EW-tLS-04C for the synthesis of perillic acid, which reached a titer of 106.69 mg/L in a 5-L fermenter. This is the first report of de novo synthesis of perillic acid in engineered microorganisms. The results also showed that other chemicals may be efficiently produced in C. tropicalis. KEY POINTS: • Key genes cytoplasmic expression was conducive to L-limonene production in C. tropicalis. • Perillic acid was first synthesized de novo in engineered microorganisms. • The titer of perillic acid reached 106.69 mg/L in a 5-L fermenter.

Innovative Analogs of Unpasteurized Kombucha Beverages: Comparative Analysis of Mint/Nettle Kombuchas, Considering Their Health-Promoting Effect, Polyphenolic Compounds and Chemical Composition.

Increasing demand for functional beverages is attracting consumers' attention and driving research to expand our knowledge of fermentation using symbiotic culture of bacteria and yeast (SCOBY) and demonstrate the health effects of consuming kombucha. The objective of this study was to develop innovative recipes for unpasteurized mint/nettle kombucha analogs, and to compare the products obtained under varying conditions in terms of chemical composition, bioactive polyphenols and health-promoting activity. Four variants of kombucha beverages (K1-K4), differing in the addition of sucrose and fermentation temperature, were formulated. The fermentation process provided data indicating the increase of antidiabetic, anti-inflammatory and anticholinergic properties, while a decrease in antioxidant capacity was observed. The content of polyphenolics was the highest on the seventh day of fermentation. A higher fermentation temperature and a larger amount of sucrose accelerated the fermentation process, which may be crucial for shortening the production time of kombucha drinks.

Chemical Characterization of the Essential Oil Compositions of Mentha spicata and M. longifolia ssp. cyprica from the Mediterranean Basin and Multivariate Statistical Analyses.

This present study aims to characterize the essential oil compositions of the aerial parts of M. spicata L. and endemic M. longifolia ssp. cyprica (Heinr. Braun) Harley by using GC-FID and GC/MS analyses simultaneously. In addition, it aims to perform multivariate statistical analysis by comparing with the existing literature, emphasizing the literature published within the last two decades, conducted on both species growing within the Mediterranean Basin. The major essential oil components of M. spicata were determined as carvone (67.8%) and limonene (10.6%), while the major compounds of M. longifolia ssp. cyprica essential oil were pulegone (64.8%) and 1,8-cineole (10.0%). As a result of statistical analysis, three clades were determined for M. spicata: a carvone-rich chemotype, a carvone/trans-carveol chemotype, and a pulegone/menthone chemotype, with the present study result belonging to the carvone-rich chemotype. Carvone was a primary determinant of chemotype, along with menthone, pulegone, and trans-carveol. In M. longifolia, the primary determinants of chemotype were identified as pulegone and menthone, with three chemotype clades being pulegone-rich, combined menthone/pulegone, and combined menthone/pulegone with caryophyllene enrichment. The primary determinants of chemotype were menthone, pulegone, and caryophyllene. The present study result belongs to pulegone-rich chemotype.

Evaluation of the effectiveness of Berberis integerrima Bunge root extract combined with spearmint essential oil in the treatment of acne vulgaris: A randomized controlled clinical trial.

BACKGROUND: Acne vulgaris is one of the most common dermatological disorders. Berberis integerrima Bunge belongs to the Berberidaceae family. Several studies on different Berberis species in addition to B. integerrima have shown antimicrobial, antioxidant, and anti-inflammatory effects. Spearmint essential oil also has antioxidant, antibacterial, and anti-inflammatory activities. This study aimed to evaluate the clinical effectiveness of the topical combination of B. integerrima root extract and spearmint essential oil in the treatment of acne vulgaris. METHODS: Patients with mild to moderate facial acne who met the inclusion criteria were randomly assigned to either drug (B. integerrima extract/spearmint essential oil topical solution) or control (clindamycin 1% topical solution) groups. Each group applied the solution twice a day for 4 weeks. Before and at the end of the intervention, the number of lesions and mGAGS (Modified Global Acne Grading Scale) score were recorded. RESULTS: Thirty patients in each group of drug and control completed the study. Topical B.integerrima root extract/spearmint essential oil significantly reduced the number of lesions (27.33 ± 26.17 vs. 21.58 ± 21.10; p < 0.001) and mGAGS (18.76 ± 8.61 vs. 13.87 ± 8.14; p < 0.001) at the end of the intervention. However, there was no significant difference between the two groups regarding the number of lesions (p = 0.906) and mGAGS (p = 0.882). CONCLUSIONS: B. integerrima root extract combined with spearmint essential oil has significant anti-acne effects, comparable to topical antibiotic clindamycin. It could be considered as a potential treatment for acne vulgaris. However, more studies with larger sample sizes and longer durations are required to confirm this effect.

Chemical composition and toxicity of commercial Mentha spicata and Eucalyptus citriodora essential oils on Culex quinquefasciatus and non-target insects.

Current vector control strategies based on synthetic chemicals are not eco-friendly against non-target organisms; hence, alternative approaches are highly required. Commercially purchased oil of Mentha spicata (Spearmint) and Eucalyptus citriodora (Citriodora) were examined against the medical pest Cx. quinquefasciatus (Say) and their non-toxicity on the aquatic species was evaluated. Chemical screening with gas chromatography coupled with mass spectrometry (GC-MS) analysis revealed a total of 14 and 11 compounds in Citriodora and Spearmint oils, respectively, with the highest peak (%) at carvone (70.44%) and isopulegol (30.4%). The larvicidal activity on the fourth instar larvae of Cx. quinquefasciatus showed dose-dependent mortality and significance at a 100 ppm concentration 48 h post-treatment with Citriodora (76.4%, P ≤ 0.001) and Spearmint (100%, P ≤ 0.001). Additionally, the photomicrograph of the fourth instar larvae revealed significant physical abnormalities in the head and midgut tissues post-exposure to Spearmint and Citriodora oils. Moreover, the histological assay revealed severe damage in the epithelial cells and gut lumen 2 to 24 h post-treatment. The repellency percentage of adult Culex mosquitoes was prominent across both oils at 150 ppm 210 min post-exposure. Non-target toxicity on the aquatic predator showed both essential oils (Spearmint oil (17.2%) and Citriodora oil (15.2%)) are safer at the maximum treatment (200 ppm) compared to temephos (75.4% at 1 ppm). The in silico screening of phyto-compounds derived by both essential oils with BeeTox (online server) showed no contact toxicity to the honey bee Apis mellifera. Overall, the present research revealed that Spearmint and Citriodora essential oils and their active phyto-compounds were toxic to Cx. quinquefasciatus and harmless to the aquatic predator and honey bee.

Simultaneous Quantification of Two Neonicotinoids Using QuEChERS-LC-MS/MS in Moroccan Spearmint (Mentha Spicata.L): Qualimetry of the Method by Uncertainty Estimation Using Generalized Pivotal Quantities Approach and Monte Carlo Simulation.

BACKGROUND: Neonicotinoids (NEOs) are used for the phytosanitary treatment of Mentha Spicata.L crops, and this practice requires precise control of these harmful substances at very low concentrations. OBJECTIVE: The objective of this study is to apply an approach allowing simultaneously validation and evaluation of measurement uncertainty based on total error methodology, in order to accurately quantify the presence of two NEOs in Mentha Spicata.L utilizing a Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS)-LC-MS/MS methodology. METHODS:  The quantification of imidacloprid and acetamiprid employing a QuEChERS extraction method, coupled with LC-MS/MS, ensuring the accuracy of the analytical method and managing the risks associated with its routine use. A complete and exhaustive validation approach based on the "ß-content, γ-confidence" tolerance interval was used for the uncertainty assessment, using the generalized pivot quantity (GPQ) concept and Monte Carlo simulation, which avoids the need for additional data while achieving intermediate precision for each concentration level within predetermined acceptable limits. RESULTS: The validation procedure is based on the choice of a quadratic model for the two NEOs, allowing the validation of acetamiprid and imidacloprid by LC-MS/MS assay within the range of working concentration. The flexibility of the uncertainty profile intervals was demonstrated with a variation in ß-content values (66.7, 80, and 90%) and risk values (10 and 5%), which remained within the acceptability limits of 20%, and the relative expanded uncertainty did not exceed 15 and 11%. CONCLUSION: A QuEChERS-LC-MS/MS method for the analysis of two NEOs has been successfully fully validated using the uncertainty profile strategy. HIGHLIGHTS: Implementation of an overall validation strategy, which involves both the validation and uncertainty assessment known as the uncertainty profile, for the quantification of two important NEOs in Mentha Spicata.L using QuEChERS-LC-MS/MS. This qualimetric approach has been conducted by computing the measurement uncertainty of the method utilizing data from analytical validation under conditions of intermediate precision at each level of concentration without additional effort. After that we have demonstrated the flexibility of this strategy for the LC-MS/MS quantification of acetamiprid and imidacloprid, using a decision tool that enables the choice and modification of ß-content and γ-confidence values.

Effects of Harvest Time and Hydrodistillation Time on Yield, Composition, and Antioxidant Activity of Mint Essential Oil.

In this study, we assessed the effects of different harvest times (9 a.m., 1 p.m., and 5 p.m.) and hydrodistillation times (60, 90, and 120 min) on the yield, chemical composition, and antioxidant activity of the spearmint (Mentha spicata L.) essential oil (EO) sourced from the Amazon region. EO yield was ≥1.55% and was not significantly influenced (p ≥ 0.05) by the different harvest times and hydrodistillation times. Thirty-one different organic compounds were identified, of which menthol (91.56-95.68%), menthone (0.6-2.72%), and isomenthone (0.55-1.46%) were the major constituents. The highest menthol content in the EO was obtained from samples collected at 9 a.m., with a hydrodistillation time of 60-90 min, compared to other harvest and hydrodistillation times. This suggests that exposure to sun and light, which is greater at harvest times of 1 p.m. and 5 p.m., decreased the menthol content and altered the chemical composition of Mentha EO. Furthermore, the sample harvested at 9 a.m. and hydrodistilled for 60 min showed the highest antioxidant activity (61.67 equivalent mg of Trolox per g of EO), indicating that antioxidant activity is strongly affected by light exposure and the contact duration of the sample with boiling water during hydrodistillation.

Modeling purple basil, sage, spearmint, and sweet basil responses to daily light integral and mean daily temperature.

Mean daily temperature (MDT) and daily light integral (DLI) can interact to influence growth and development of plants. Our objectives were to determine 1) the extent DLI and MDT influence growth and development of purple basil 'Dark Opal' (Ocimum basilicum), sage 'Extrakta' (Salvia officinalis), spearmint 'Spanish' (Mentha spicata), and sweet basil 'Nufar' (Ocimum basilicum) and 2) the influence on purple basil color. Young plants were transplanted into hydroponic systems in five greenhouse compartments with MDT set points of 23, 26, 29, 32, or 35°C and DLIs from 5 to 19 mol·m‒2·d‒1, respectively. At harvest, growth, development, and leaf color was measured. Branch number of all genera increased as MDT increased from ~23 to 35°C. Sweet basil branch number increased as DLI increased from 5.5 to 13.2 mol·m‒2·d‒1, but the effect of DLI was attenuated as MDT decreased. In contrast, increasing DLI from ~5-6 to ~18-19 mol·m‒2·d‒1 increased sage and spearmint branch number more when MDT was lower (~23°C) compared to ~35°C, while branch number of purple basil was not influenced by DLI. The optimal MDT (MDTopt) for sage and spearmint fresh mass decreased from 27.5 to 23.5°C and from 30.4 to 27.8°C, respectively, as DLI increased from 6 to 18 mol·m‒2·d‒1, while sweet basil fresh mass MDTopt increased from 32.6 to 35.5°C as DLI increased from 6 to 11 mol·m‒2·d‒1. Purple basil was greener [hue angle (h°) = 99° to 138°] when MDT was ~35°C regardless of DLI, but when MDT was lower (~25°C), basil was more purple (h° = 335°) at a DLI of 18.7 compared to 5.0 mol·m‒2·d‒1 (h° = 98°). Taken together, MDT and DLI can have a large impact on plant growth, development, and color and can be manipulated to achieve desired characteristics.

Study on Antioxidant Activity of Spearmint Essential Oil and Pure Dew.

Spearmint essential oil and pure dew were used as research objects, the antioxidant capacity of spearmint was evaluated by measuring the scavenging capacity of superoxide anion radical and hydroxyl radical, providing technical support for the subsequent development and utilization of spearmint truffle and essential oil. The results showed that when the volume fraction (V/V) of spearmint essential oil was 1%, its antioxidant capacity was the strongest, and its scavenging rates of superoxide anion radical and hydroxyl radical were 50.94% and 90.11% respectively; When the volume fraction (V/V) of spearmint hydrosol was 100%, its antioxidant capacity was the strongest, and its scavenging rates of superoxide anion radical and hydroxyl radical were 47.65% and 45.60%.

Efficacy of a Spearmint (Mentha spicata L.) Extract as Nutritional Support in a Rat Model of Hypertensive Glaucoma.

Purpose: Glaucoma is an eye-brain axis disorder characterized by loss of retinal ganglion cells (RGCs). Although the role of intraocular pressure (IOP) elevation in glaucoma has been established, the reduction of oxidative stress and inflammation has emerged as a promising target for neuronal tissue-supporting glaucoma management. Therefore, we evaluated the effect of a proprietary spearmint extract (SPE) on RGC density, activity, and neuronal health markers in a rat model of hypertensive glaucoma. Methods: Animals were divided in four groups: untreated healthy control and three glaucomatous groups receiving orally administered vehicle, SPE-low dose, or SPE-high dose for 28 days. Ocular hypertension was induced through intracameral injection of methylcellulose at day 15. At day 29, rats underwent electroretinogram (ERG) recordings, and retinas were analyzed for RGC density and markers of neural trophism, oxidative stress, and inflammation. Results: SPE exerted dose-dependent response benefits on all markers except for IOP elevation. SPE significantly improved RGC-related ERG responses, cell density, neurotrophins, oxidative stress, and inflammation markers. Also, in SPE-high rats, most of the parameters were not statistically different from those of healthy controls. Conclusions: SPE, a plant-based, polyphenolic extract, could be an effective nutritional support for neuronal tissues. Translational Relevance: These results suggest that SPE not only may be a complementary approach in support to hypotensive treatments for the management of glaucoma but may also serve as nutritional support in other ocular conditions where antioxidant, anti-inflammatory, and neuroprotective mechanism are often disrupted.

Green synthesis, characterization, and biological evaluation of gold and silver nanoparticles using Mentha spicata essential oil.

Green synthesis of bioactive nanoparticles (NPs) is getting more attractive in various fields of science including the food industry. This study investigates the green synthesizing and characterization of gold NPs (AuNPs) and silver NPs (AgNPs) produced using Mentha spicata L. (M. spicata) essential oil as well as their antibacterial, antioxidant, and in vitro cytotoxic effects. The essential oil was mixed with both Chloroauric acid (HAuCl4) and aqueous silver nitrate (AgNO3) solutions separately and incubated at room temperature for 24 h. The chemical composition of the essential oil was identified by gas chromatography coupled with a mass spectrometer detector (GC-MS). Au and Ag nanoparticles were characterized using UV-Vis spectroscopy, transmission electron microscopy, scanning electron microscopy, dynamic light scattering (DLS), X-ray diffraction (XRD) and Fourier transform infrared (FTIR). The cytotoxicity of both types of nanoparticles was evaluated using MTT assay on cancerous HEPG-2cell line by exposing them to various concentrations of both NPs for 24 h. The antimicrobial effect was evaluated by the well-diffusion technique. The antioxidant effect was determined by DPPH and ABTS tests. According to the results of GC-MS analysis, 18 components were identified, including carvone (78.76%) and limonene (11.50%). UV-visible spectroscopy showed a strong absorption peak of 563 nm and 485 nm, indicating the formation of Au NPs and Ag NPs, respectively. TEM and DLS demonstrated that AuNPs and AgNPs were predominantly spherical shaped with average sizes of 19.61 nm and 24 nm, respectively. FTIR analysis showed that biologically active compounds such as monoterpenes could assist in the formation and stabilization of both types of NPs. Additionally, XRD provided more accurate results, revealing a nano-metal structure. Silver nanoparticles exhibited better antimicrobial activity against the bacteria than AuNPs. Zones of inhibition ranging 9.0-16.0 mm were recorded for the AgNPs, while zones of 8.0-10.33 mm were observed AuNPs. In the ABTS assay, the AuNPs and AgNPs showed a dose-dependent activity and synthesized nanoparticles exhibited higher antioxidant activity than MSEO in both assays. Mentha spicata essential oil can be successfully used for the green production of Au NPs and Ag NPs. Both green synthesized NPs show antibacterial, antioxidant, and in vitro cytotoxic activity.

Green nanotechnology advances: green manufacturing of zinc nanoparticles, characterization, and foliar application on wheat and antibacterial characteristics using Mentha spicata (mint) and Ocimum basilicum (basil) leaf extracts.

Due to their distinctive characteristics and widespread application across all scientific disciplines, nanoparticles have attracted a lot of attention in the current millennium. Green synthesis of ZnO-NPs is gaining a lot of interest at the moment due to a number of its advantages over traditional methods, including being quicker, less expensive, and more environmentally friendly. In the current study, two distinct plant extracts are used to quickly, cheaply, and environmentally friendly synthesize zinc oxide nanoparticles (ZnO-NPs). Mint (Mentha spicata) and basil (Ocimum basilicum) were the plants employed in this study as stabilizing agents to synthesize ZnO-NPs with a green chemistry approach. The innovative aspect of the study is the use of mint and basil extracts in the conversion of zinc chloride to zinc oxide and then determining the effect of these two types of nanoparticles produced by green synthesis on the growth parameters of the plant when they reach the plants by foliar spraying and their uptake by plants and evaluating the antibacterial properties of these nanoparticles. The physical properties of the produced nanoparticles were investigated using XRD, SEM, and FTIR. Moreover, Escherichia coli and Staphylococcus aureus were used to demonstrate the antibacterial properties of ZnO-NPs against both gram-positive and gram-negative bacteria, respectively. Synthesized ZnO-NPs were also given as foliar treatment in order to determine Zn+2 uptake by plants and potential toxic effects on the growth of wheat. The shape of ZnO-NPs was triangular, as revealed by SEM analysis. In the X-ray diffraction study, strong and clearly discernible sharp peaks were seen, with an average size of 24.5 nm for M-ZnO-NPs and 26.7 nm for B-ZnO-NPs determined using Scherrer's formula. The phytoconstituents of the plant extract served as capping/stabilizing agents during the synthesis of ZnO-NPs, as demonstrated by Fourier transform-infrared spectroscopy. The produced nanoparticles were applied to the green parts of wheat plants by spraying, and the development of the plants and the change of zinc uptake were investigated. At the same time, the effect of these three types of nanoparticles on the germination of wheat seeds in the soil medium containing these nanoparticles was investigated. According to experimental results, M-ZnO-NPs (produced from mint) and B-ZnO-NPs (produced from basil) improved the germination percentage of wheat at 400 mg/L concentration (100%), while raw ZnO-NPs showed 90% germination at the same concentration. When the Zn+2 uptake of the plant by the leaves depending on the Zn+2 concentration in the environment after spraying was examined, it was determined that the Zn+2 uptake of the plants increased due to the increase in the applied Zn+2 concentration. The highest Zn+2 uptake of the plant was determined as 50, 25, and 50 mg/L for M-ZnO-NP, B-ZnO-NPs, and raw ZnO-NPs, respectively. Therefore, it has been determined that plant growth varies depending on the type and concentration of ZnO-NPs, and therefore, if foliar nanoparticle applications are made to wheat, the threshold concentrations, sizes, and types of ZnO-NPs should be carefully evaluated. In addition, antibacterial properties results showed that S. aureus was more sensitive to all three types of ZnO-NPs than E. coli.

Prediction of essential oil content in spearmint (Mentha spicata) via near-infrared hyperspectral imaging and chemometrics.

Spearmint (Mentha spicata L.) is grown for its essential oil (EO), which find use in food, beverage, fragrance and other industries. The current study explores the ability of near infrared hyperspectral imaging (HSI) (935 to 1720 nm) to predict, in a rapid, nondestructive manner, the essential oil content of dried spearmint (0.2 to 2.6% EO). Spectral values of spearmint samples varied considerably with spatial coordinates, and so the use of averaging the spectral values of a surface scan was warranted. Data preprocessing was done with Multiplicative Scatter Correction (MSC) or Standard Normal Variate (SNV). Selection of spectral input variables was done with Least Absolute Shrinkage and Selection Operator (LASSO), Principal Component Analysis (PCA) or Partial Least Squares (PLS). Regression was executed with linear regression (LASSO, PLS regression, PCA regression), Support Vector Machine (SVM) regression, and Multilayer Perceptron (MLP). The best prediction of EO concentration was achieved with the combination of MSC or SNV preprocessing, PLS dimension reduction, and MLP regression (1 hidden layer with 6 nodes), achieving a good prediction with a ratio of performance to deviation (RPD) of 2.84 ± 0.07, an R2 of prediction of 0.863 ± 0.008, and a RMSE of prediction of 0.219 ± 0.005% EO. These results show that NIR-HSI is a viable method for rapid, nondestructive analysis of EO concentration. Future work should explore the use of NIR in the visible spectrum, the use of HSI for determining EO in other plant materials and the potential of HSI to determine individual compounds in these solid plant/food matrices.

The effect of irrigation with treated and untreated wastewater on the yield and chemical composition of essential oil of Mentha spicata L. and Rosmarinus officinalis L.

Today, the lack of quality water supply has led to the tendency to use unconventional water to irrigate agricultural products. Considering the importance and application of essential oils of mint plants in various pharmaceutical, food, and health industries and also considering the approach of using unconventional waters in the cultivation of medicinal plants, the present study aimed to investigate and compare the chemical composition of essential oils of two species of Mentha spicata L. and Rosmarinus officinalis L. which was designed and implemented for the first time under the influence of different treatments of municipal and industrial wastewater. For this purpose, first R. officinalis cuttings and roots of M. spicata were prepared and after preparing and leveling the ground, in the spring of 2020, it was transferred to the planting site and planted in the form of creek and ridges. The treatments studied in this study included well water (WW), treated municipal wastewater (TMW), untreated municipal wastewater (UMW), treated industrial wastewater (TIW), and untreated industrial wastewater (UIW) in a randomized complete block design with four repeat runs. After watering the plants continuously for 3 months, the plant branches were collected and transferred to the laboratory for drying. After extracting the essential oil by water distillation (Clevenger) method, the analysis and identification of the compounds were performed by a chromatograph coupled with a mass spectrometer (GC/MS). The results showed that the highest and lowest yields of M. spicata belonged to the samples treated with UMW and WW, respectively. Also, R. officinalis essential oil irrigated with UMW and UIW had the highest and lowest yields, respectively. The number of essential oil compounds in of M. spicata was between 5 and 19 and in R. officinalis between 14 and 23 under different treatments. The results of the analysis of essential oil compounds showed that D-carvone (57.77-57.44%) and D-limonene (8.70-26.65%) for M. spicata and α-pinene (26.12-34.85%), 1,8-cineole (18.95-23.70%), and camphene (9.93-12.80%) for R. officinalis were predominant compounds in all studied treatments. The results show that UMW is a suitable and efficient treatment to have the best quantity of M. spicata essential oil and the best quality and quantity of R. officinalis essential oil.

Interactions of Chitosan-coated Green Synthesized Silver Nanoparticles using Mentha spicata and Standard Antibiotics against Bacterial Pathogens.

BACKGROUND: Infectious diseases are caused by various multidrug-resistant pathogenic bacteria and in recent scenarios, nanoparticles have been used as innovative antimicrobial agents. AIMS: This current research aimed to evaluate the bactericidal effect of chitosan-coated green synthesized silver nanoparticles using aqueous extract of Mentha spicata (MSaqu) against bacterial pathogens, i.e., Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Serratia marcescens, Staphylococcus aureus, and Streptococcus pyogenes. METHODS: Synthesis and characterization of silver nanoparticles (MSAgNPs) were carried out via atomic absorption spectrometer and Fourier-transform infrared spectroscopy. Agar well and agar disc diffusion methods were used to assess the antibacterial and synergistic effect of chitosanmediated biogenic silver nanoparticles and standard antibiotics. Three types of interactions, i.e., antagonistic (↓), synergistic (↑), and additive (¥) were observed. RESULTS: Synergistic effect was recorded against Pseudomonas aeruginosa (8.5±0.25 mm↑), Serratia marcescens (19.0±1.0 mm↑), and Klebsiela pneumonia (8.5±0.25 mm↑), an additive effect was exhibited by Escherichia coli (9.0±0.0 mm¥), Streptococcus pyogenes (10.0±0.0 mm¥), and Staphylococcus aureus (7.5±0.25 mm↓) and they showed antagonistic effects when chitosan-coated silver nanoparticles (CLMSAgNPs) were applied compared to chitosan, MSaqu, and MSAgNPs. Interesting antibacterial results were recorded when chitosan-coated Mentha spicata extract and silver nanoparticles were applied along with antibiotics. The synergistic effects of chitosan-coated silver nanoparticles (CLMSAgNPs) + K were recorded against E. coli (14.5±0.25 mm). The synergistic effects of chitosan-coated silver nanoparticles (CLMSAgNPs) + AML were recorded against E. coli (5.5±0.0 mm), S. pyogenes (10.0±0.0 mm), K. pneumonia (5.5±0.0 mm), and S. aureus (4.0±0.0 mm). The synergistic effects of chitosan-coated silver nanoparticles (CLMSAgNPs) + NOR were recorded against E. coli (16.0±0.0 mm), P. aeruginosa (19.0±0.0 mm), S. marcescens (19.5±0.25 mm), S. pyogenes (11.5.0±0.25 mm), K. pneumonia (23.0±0.0 mm), and S. aureus (8.5±0.25 mm). CONCLUSION: Current findings concluded that chitosan-coated biogenic silver nanoparticles have potential bactericidal effects against infectious pathogens and could be used as forthcoming antibacterial agents.

Mitigation of salinity impact in spearmint plants through the application of engineered chitosan-melatonin nanoparticles.

High soil salinity represents a critical environmental constraint to crop production. In order to ameliorate the effects of salinity, a plethora of molecules have been applied and promising outcomes have been noted. The beneficial effects of chitosan (CTS) and melatonin (Mel) application, separately, have been previously recorded with respect to plant growth and productivity, leading to the hypothesis that their conjugation in the form of chitosan-melatonin nanoparticles (CTS-HPMC-Mel NPs) could lead to further enhanced performance of plants under control and stress conditions. In this regard, novel CTS-HPMC-Mel NPs were synthesized, characterized and then employed as a chemical priming agent in spearmint (Mentha spicata L.) plants 24 h prior to salinity stress imposition. As expected, salt stress negatively affected morphophysiological attributes such as plant height, leaf number, leaf fresh weight, leaf dry weight, photosynthetic pigments, Fv/Fo, and Fv/Fm. On the other hand, stress-related attributes, such as content of proline, MDA and H2O2, as well as activity of APX and GP enzymes were increased in response to salt stress. However, adverse effects of salt stress were ameliorated with Mel and CTS-HPMC-Mel NP treatments by enhancing morphological traits, proline, antioxidant enzymatic activities, as well as content of dominant constituents of essential oil profile. It is worth noting that conjugated form of Mel with chitosan, in comparison with solo treatment of Mel, was more effective in combating stress effects. To our knowledge, this is the first report to demonstrate that engineered CTS-HPMC-Mel NPs could be applied as an innovative protective agent to mitigate the effects of salinity in crop plants.

δ18O compound-specific stable isotope assessment: An advanced analytical strategy for sophisticated adulterations detection in essential oils - Application to spearmint, cinnamon, and bitter almond essential oils authentication.

Natural plant extracts are primarily used as raw materials in the cosmetic and perfumery industry. However, adulterations with petrochemical products are occurring in the market, leading to non-100% natural products. Several analytical techniques such as impurity detection or enantioselective ratio assessments have been previously described as good indicators to detect any addition of synthetic products, but these techniques are ineffective with novel type of synthetic pathways such as semisynthesis. In order to improve authentication, development of advanced analytical strategies such as δ18O stable isotopic ratios assessment was tested on spearmint, cinnamon and bitter almond essential oils major metabolites (carvone, (E)-cinnamaldehyde, and benzaldehyde). Natural δ18O mean values (δ18OCarvone = 18.4‰; δ18OCinnamaldehyde = 13.9‰; δ18OBenzaldehyde = 16.5‰) were found to be higher than semisynthetic origin for the 3 studied molecules (δ18OCarvone = 9.2‰; δ18OCinnamaldehyde = 8.8‰; δ18OBenzaldehyde = 10.9‰). These measurements proved to be efficient to discriminate natural and semisynthetic origins of these components and therefore potentially lead to a novel way to authenticate natural products.

Bioactive Profile of Distilled Solid By-Products of Rosemary, Greek Sage and Spearmint as Affected by Distillation Methods.

By-products of essential oils (EOs) in the industry represent an exploitable material for natural and safe antioxidant production. One representative group of such by-products is distilled solid residues, whose composition is properly modulated by the distillation method applied for the recovery of EOs. Recently, in terms of Green Chemistry principles, conventional extraction and distillation processes are considered outdated and tend to be replaced by more environmentally friendly ones. In the present study, microwave-assisted hydro-distillation (MAHD) was employed as a novel and green method for the recovery of EOs from three aromatic plants (rosemary, Greek sage and spearmint). The method was compared to conventional ones, hydro-distillation (HD) and steam-distillation (SD), in terms of phytochemical composition of distilled solid residues, which was estimated by spectrophotometric and chromatographic methods. Total phenolic content (TPC), total flavonoid content (TFC) and antioxidant activity (ABTS, DPPH and FRAP) results highlighted the distilled solid residues as good sources of antioxidants. Moreover, higher antioxidant activity was achieved for MAHD extracts of solid residues in comparison to HD and SD extracts. A metabolomics approach was carried out on the methanolic extracts of solid residues obtained by different distillation methods using LC-MS analysis followed by multivariate data analysis. A total of 29 specialized metabolites were detected, and 26 of them were identified and quantified, presenting a similar phenolic profile among different treatments, whereas differences were observed among different species. Rosmarinic acid was the most abundant phenolic compound in all extracts, being higher in MAHD extracts. In rosemary and Greek sage extracts, carnosol and carnosic acid were quantified in significant amounts, while trimers and tetramers of caffeic acid (salvianolic acids isomers) were identified and quantified in spearmint extracts, being higher in MAHD extracts. The obtained results pointed out that MAHD extracts of distilled solid by-products could be a good source of bioactives with potential application in the food, pharmaceutical and cosmetic industries, contributing to the circular economy.

Antioxidative, anticancer, and antibacterial activities of a nanogel containing Mentha spicata L. essential oil and electrospun nanofibers of polycaprolactone-hydroxypropyl methylcellulose.

BACKGROUND: As the largest organ, the skin has been frequently affected by trauma, chemical materials, toxins, bacterial pathogens, and free radicals. Recently, many attempts have been made to develop natural nanogels that, besides hydrating the skin, could also be used as antioxidant or antibacterial agents. METHODS: In this study, the chemical composition of the Mentha spicata essential oil was first investigated using GC-MS analysis. Its nanoemulsion-based nanogel was then investigated; successful loading of the essential oil in the nanogel was confirmed using FTIR analysis. Besides, nanogel's antioxidative, anticancer, and antibacterial activities were investigated. RESULTS: Carvone (37.1%), limonene (28.5%), borneol (3.9%), ß-pinene (3.3%), and pulegone (3.3%) were identified as five major compounds in the essential oil. By adding carboxymethylcellulose (3.5% w/v) to the optimal nanoemulsion containing the essential oil (droplet size of 196 ± 8 nm), it was gelified. The viscosity was fully fitted with a common non-Newtonian viscosity regression, the Carreau-Yasuda model. The antioxidant effect of the nanogel was significantly more potent than the essential oil (P < 0.001) at all examined concentrations (62.5-1000 µg/mL). Furthermore, the potency of the nanogel with an IC50 value of 55.0 µg/mL was substantially more (P < 0.001) than the essential oil (997.4 µg/mL). Also, the growth of Staphylococcus aureus and Escherichia coli after treatment with 1000 µg/mL nanogel was about 50% decreased compared to the control group. Besides, the prepared electrospun polycaprolactone-hydroxypropyl methylcellulose nanofibers mat with no cytotoxic, antioxidant, or antibacterial effects was proposed as lesion dressing after treatment with the nanogel. High potency, natural ingredients, and straightforward preparation are advantages of the prepared nanogel. Therefore, it could be considered for further consideration in vivo studies.