Assessment and Optimization of the Insecticidal Properties of γ-Al2O3 Nanoparticles Derived from Mentha pulegium By-Products to Xylosandrus crassiusculus (Carob Beetle).

This study concentrates on assessing the insecticidal attributes of the γ-Al2O3 nanoparticles derived from the remnants of Mentha pulegium, which include essential oil, ethanolic extract, and plant waste. The synthesis of the γ-Al2O3 nanoparticles was executed using a direct sol-gel procedure, affirming the crystal structure according to extensive physicochemical analyses such as UV-Vis, XRD, FTIR, and SEM. Evaluation of the insecticidal activity in vitro was conducted against Xylosandrus crassiusculus, a pest that infests carob wood, utilizing strains from diverse forests in the Khenifra region, situated in the Moroccan Middle Atlas. The lethal doses 50 ranged from 40 mg/g to 68 mg/g, indicating moderate effectiveness compared to the commercial insecticide Permethrin. Optimization of the conditions for the efficiency of the γ-Al2O3 nanoparticles was determined using experimental plans, revealing that time, humidity, and temperature were influential factors in the lethal dose 50 of these nanomaterials. Moreover, this study encompasses the establishment of correlations using Principal Component Analysis (PCA) and Ascending Hierarchical Classification (AHC) among various geographic, biological, and physical data, amalgamating geographic altitude and γ-Al2O3 nanoparticle insecticide parameters, as well as the attributes of the mechanical tests conducted on the carob wood affected by insects. The correlations highlight the close connections between the effectiveness of the insecticide, mountain altitude, and the mechanical parameters that were examined. Ultimately, these nanoparticles demonstrate promising potential as alternative insecticides, thus opening up encouraging prospects for safeguarding against carob wood pests.

In vitro ruminal fermentation, core nutrient, fatty acids and mineral matter of pennyroyal (Mentha pulegium L.) herbage at different phenological stages.

BACKGROUND: In ruminants, fibrous feedstuffs must be included in the ration to ensure normal rumen physiology and to prevent the occurrence of rumen-related metabolic diseases. In addition to being a source of fibrous feedstuffs, they contain energy depending on the level of digestion and protein, minerals, fatty acids, minerals, and secondary compounds. OBJECTIVES: This study aimed to determine the nutrient matter, fatty acid, mineral and in vitro rumen fermentation values of the pennyroyal (Mentha pulegium L.) plant. METHODS: The pennyroyal plant samples were collected at different phenological stages (vegetative, full flowering, and seed bulking) from the natural meadow. The samples were analysed for core nutrients, condensed tannins, minerals, fatty acids, and in vitro ruminal fermentation parameters. RESULTS: The calcium (Ca) and iron (Fe) contents and in vitro ruminal fermentation parameters (total gas production and methane production, organic matter digestion (OMd), and the ammonia-nitrogen) decreased with increasing phenological stage (p < 0.05). The percentages of linoleic, α-linolenic, ω-3, ω-6 and polyunsaturated fatty (PUFA) acids of the pennyroyal plant linearly increased with the phenological stages (p < 0.05). However, butyric acid (BA) concentration in the in vitro ruminal fermentation fluid in the full flowering stage was lower than that of other stages (p < 0.05). CONCLUSIONS: Pennyroyal plant is a functional plant in terms of high values of ether extract (EE), α-linolenic acid, linoleic acid, ∑ω-3 fatty acids, Ca, Fe and Zn contents. For this plant to be used as animal feed, the stage when it has the highest values for Ca, Mg, S and Zn minerals and in vitro OMd were vegetative and full flowering. The stage with good potential as animal feed for ∑ω-3 and ∑ω-6 fatty acids and core nutrients (CP and EE) is the seed bulking stage.

Radioprotective effect of nanoniosome loaded by Mentha Pulegium essential oil on human peripheral blood mononuclear cells exposed to ionizing radiation.

OBJECTIVE: The present study aimed to assess the radioprotective effect of nanoniosomes loaded by Mentha Pulegium essential oil (MPEO-N nanoparticles) as a natural antioxidant on human peripheral blood mononuclear cells (PBMCs). SIGNIFICANCE: Despite the applications and advantages of ionizing radiation, there are many radiation risks to biological systems that are necessary to be reduced as much as possible. METHODS: MPEO-N nanoparticles were prepared by the lipid thin film hydration method, and its physicochemical characteristics were analyzed. PBMCs were then irradiated with X-ray using a 6 MV linear accelerator at two radiation doses in the presence of nontoxic concentrations of MPEO-N nanoparticles (IC10). After 48 and 72 h of incubation, the radioprotective effect was investigated by measuring survival, apoptosis, and necrosis of PBMCs, using MTT assay and flow cytometry analysis. KEY FINDINGS: The hydrodynamic diameter and zeta potential of nanoniosomes were 106.0 ± 4.69 nm and -15.2 ± 0.9 mV, respectively. The mean survival percentage of PBMCs showed a significant increase only at a radiation dose of 200 cGy compared with the control group. The percentages of apoptosis and necrosis of cells in the presence of MPEO-N nanoparticles at both radiation doses and incubation periods (48 and 72 h) demonstrated a significant reduction compared with the control. CONCLUSION: MPEO-N nanoparticles as a natural antioxidant, exhibited a favorable radioprotective effect by a significant reduction in the percentage of apoptosis and necrosis of irradiated PBMCs.

Different cultivation systems and foliar application of calcium nanoparticles affect the growth and physiological characteristics of pennyroyal (Mentha pulegium L.).

The aim of this study was to investigate the impact of different cultivation systems (soil cultivation, hydroponic cultivation in greenhouse conditions, and hydroponic vertical cultivation in plant factory under different LED lights) and foliar spraying of nano calcium carbonate on pennyroyal plants. Nano calcium carbonate was applied to the plants at a 7-day interval, three times, one month after planting. Results showed that the greenhouse cultivation system with calcium carbonate foliar spraying produced the highest amount of shoot and root fresh mass in plants. Additionally, foliar spraying of calcium carbonate increased internode length and leaf area in various cultivation systems. Comparing the effects of different light spectrums revealed that red light increased internode length while decreasing leaf length, leaf area, and plant carotenoids. Blue light, on the other hand, increased the leaf area and root length of the plants. The hydroponic greenhouse cultivation system produced plants with the highest levels of chlorophyll, carotenoids, and phenolic compounds. White light-treated plants had less iron and calcium than those exposed to other light spectrums. In conclusion, pennyroyal plants grown in greenhouses or fields had better growth than those grown in plant factories under different light spectrums. Furthermore, the calcium foliar application improved the physiological and biochemical properties of the plants in all the studied systems.

Satureja montana and Mentha pulegium essential oils' antimicrobial properties against Pseudomonas syringae pv. actinidiae and elicitor potential through the modulation of kiwifruit hormonal defenses.

Pseudomonas syringae pv. actinidiae (Psa) is responsible for the kiwifruit bacterial canker, the most severe disease of Actinidia spp. The use in agriculture of antibiotics and cooper-based compounds is increasingly being restricted, demanding for new sustainable alternatives to current agrochemicals. We aimed to characterize the anti-Psa potential of essential oils (EOs) of Mentha pulegium and Satureja montana and investigate if they elicit the plant-host hormonal defenses. The EOs were characterized through gas-chromatography with flame ionization detector (GC-FID) and mass spectrometry (MS). Pulegone (78.6%) and carvacrol (43.5%) were the major constituents of M. pulegium and S. montana EO, respectively. Only S. montana EO showed relevant anti-Psa activity in vitro. To evaluate if the EOs also elicited host defenses, in vitro shoots were treated with 2 mg shoot-1 of EO-solution and subsequently inoculated with Psa three days later. Shoots were analyzed 10 min, three days (and 10 min after Psa-inoculation), four and ten days after EO application. The up/down regulation of RNA-transcripts for hormone biosynthesis, Psa biofilm production and virulence genes were quantified by real-time quantitative PCR (RT-qPCR). Phytohormones were quantified by High-Performance Liquid Chromatography (HPLC). S. montana EO showed the most promising results as a defense elicitor, increasing 6-benzylaminopurine (BAP) by 131.07% and reducing indole-3-acetic acid (IAA) levels by 49.19%. Decreases of salicylic acid (SA), and gibberellic acid 3 (GA3) levels by 32.55% and 33.09% respectively and an increase of abscisic acid (ABA) by 85.03%, in M. pulegium EO-treated shoots, revealed some protective post-infection effect. This is the most comprehensive research on the Psa's impact on phytohormones. It also unveils the protective influence of prior EO exposure, clarifying the plant hormonal response to subsequent infections. The results reinforce the hypothesis that carvacrol-rich S. montana EO can be a suitable disease control agent against Psa infection. Its dual action against pathogens and elicitation of host plant defenses make it a promising candidate for incorporation into environmentally friendly disease management approaches. Nonetheless, to fully leverage these promising results, further research is imperative to elucidate the EO mode of action and evaluate the long-term efficacy of this approach.

Variability in Chemical Composition and Antimicrobial Activity of Mentha pulegium L. Essential Oil, Cultivated under Different Plant Growth Promoting Rhizobacteria.

The aim of present work was to study the essential oil chemical composition and antimicrobial activity of cultivated Mentha pulegium L. under different plant growth promoting rhizobacteria (Pseudomonas fluorescens, Bradyrhizobium sp. and Sinorhizobium meliloti) individually and in consortium. Yield, in plants inoculated with Bradyrhizobium sp. and S. meliloti in consortium, increase significantly relative to control plants. GC and GC/MS analyses pointed to a qualitative and quantitative variability of components. The investigated essential oils were clustered into three chemotypes: piperitenone/1,8-cineol (40.9/29.4 %) chemotype in plants inoculated with Bradyrhizobium sp. individually, S. meliloti individually, and Bradyrhizobium sp. and S. meliloti in consortium, piperitone/menthone (41.8/33.8 %) chemotype in plants inoculated with P. fluorescens individually, P. fluorescens and Bradyrhizobium sp. in consortium, and P. fluorescens and S. meliloti in consortium and pulegone/menthol (47.9/31.5 %) chemotype in control plants. The antimicrobial activity, carried out by the disc diffusion method and the determination of the Minimum Inhibitory Concentration (MIC) against ten microorganisms, varied significantly according to the tested microorganism and the rhizobacterial species used individually or in consortium (inhibition zone: 8.5-33.5 mm; MIC: 0.25-2.5 µL/mL). Our findings provided useful indications to select interesting chemotype within M. pulegium, especially in perspective of its cultivation.

Nano-scaled emulsion and nanogel containing Mentha pulegium essential oil: cytotoxicity on human melanoma cells and effects on apoptosis regulator genes.

BACKGROUND: Topical drug delivery using nanoemulsions and nanogels is a promising approach to treating skin disorders such as melanoma. METHODS: In this study, the chemical composition of Mentha pulegium essential oil with five major compounds, including pulegone (68.11%), l-menthone (8.83%), limonene (2.90%), iso-pulegone (2.69%), and iso-menthone (1.48%) was first identified using GC-MS (Gas chromatography-Mass Spectrometry) analysis. Afterward, a nano-scaled emulsion containing the essential oil with a droplet size of 7.70 ± 1 nm was prepared. Nanogel containing the essential oil was then prepared by adding (2% w/v) carboxymethyl cellulose to the nano-scaled emulsion. Moreover, the successful loading of M. pulegium essential oil in the nano-scaled emulsion and nanogel was confirmed using ATR-FTIR (Attenuated total reflectance-Fourier Transform InfraRed) analysis. Then, human A375 melanoma cells were treated with different concentrations of samples, the MTT assay evaluated cell viability, and cell apoptosis was confirmed by flow cytometry. In addition, the expression of apoptotic and anti-apoptotic genes, including Bax and Bcl-2, was evaluated using the qPCR (quantitative Polymerase Chain Reaction) technique. RESULTS: The results showed that cell viability was reduced by 90 and 45% after treatment with 300 µg/mL of the nanogel and nano-scaled emulsion. As confirmed by flow cytometry, this effect was mediated by apoptosis. Furthermore, gene expression analysis showed up-regulation of Bax and down-regulation of Bcl-2 genes. Therefore, the prepared nanogel, with high efficacy, could be considered a potent anticancer agent for supplementary medicine and in vivo research.

Effects of Pennyroyal (Mentha pulegium L.) Supplementation on production performance, egg quality traits, and biochemical parameters of blood and egg in laying hens at later stages of the production period.

BACKGROUND: According to the promising outcomes acquired in recent studies that focused on using natural feed additives in animal nutrition, poultry nutritionists have also been interested in investigating these medicinal herbs' effects on poultry performance and egg characteristics. Pennyroyal (Mentha pulegium L.), a natural source of antioxidant, antimicrobial, anti-inflammatory, etc. compounds, is recommended as a potential alternative for antibiotics, with similar benefits and no adverse effects on animal health or their products. Considerable effects have been reported on laying hens' performance and their eggs' internal and external traits by pennyroyal inclusion in their diets. OBJECTIVES: Due to the different results reported by pennyroyal inclusion in laying hens' diets in recent limited studies, this study was designed to reinvestigate the impacts of pennyroyal (Mentha pulegium L.) supplementation (PS) on productive performance, egg quality traits, and biochemical compounds in blood and egg yolk in aged laying hens. The findings of this experiment may help for a better understanding of PS impacts on desired parameters, along with improving aged laying hens' performance. MATERIALS AND METHODS: A total of 144 Leghorn (Hyline-W36 strain) laying hens (53 weeks old) were used to investigate the effects of PS on performance, egg quality, and biochemical parameters of blood and egg yolk in old laying hens. The animals were distributed into three treatment groups; including a control diet (without PS), 0.5%, and 1% PS diets with six replicates and eight birds per replicate. RESULTS: The results indicated that the egg production rate (except for week eight) and egg mass were significantly reduced by PS in the laying hens' diet during the first four weeks and the total period of the experiment (P<0.05). However, the feed conversion ratio increased by PS (P<0.05). Based on the egg quality trait evaluations (week 4), the yolk percentage, yolk height, and yolk index reduced, while shell-breaking strength represented a significant increase in PS groups (P<0.05). However, shell thickness and albumen percentage reduced and increased by 1% PS, respectively (P<0.05). At week eight, the Haugh unit reduced while the albumen pH increased by PS (P<0.05). The findings revealed a decline in the egg density in 1% PS groups (P<0.05). In the case of serum biochemical parameters, alanine aminotransferase increased, whereas malondialdehyde (MDA) levels reduced in the PS groups (P<0.05). Moreover, the triglyceride levels of both serum and egg yolk (accompanied by the very low-density lipoprotein level of serum) decreased by PS (P<0.05). CONCLUSIONS: In conclusion, PS reduced serum and egg yolk triglyceride levels, accompanied by serum MDA, along with reducing production performance.

Formulation of nanoemulsion carriers containing Pennyroyal (Mentha pulegium) and Gijavash (Froriepia subpinnata) essential oils for enriching Doogh (Iranian dairy drink).

Today, the increasing use of chemical preservatives in foods is considered one of the main problems in food industries. This study aimed to produce the pasteurised Doogh (Iranian yogurt drink) containing a nanoemulsion of essential oil (EO) with appropriate quality. A factorial test based on a completely randomised design with two treatments in three levels, including EO type (pennyroyal, Gijavash, and their equal combination) and a control sample was applied to assess the physicochemical and sensory properties of Doogh. The highest negative zeta potential and antioxidant activity percentage were observed in the sample containing the nanoemulsion of pennyroyal and enriched with a combination of two essential oils. The microbial evaluation results indicated that the total microorganism count was minimised in the Doogh containing the nanoemulsion of Gijavash. The nanoemulsions of pennyroyal and Gijavash can be added into Doogh formulation to produce a new product with maximum sensory acceptability.

Characterization of essential oils and volatiles from the aerial parts of Mentha pulegium L. (Lamiaceae) using microwave-assisted hydrodistillation (MAHD) and headspace solid phase microextraction (HS-SPME) in combination with GC-MS.

Herein, the chemical profiles of the essential oils and volatiles obtained from the aerial parts of Mentha pulegium L. (Lamiaceae) are reported respectively using microwave-assisted hydrodistillation (MAHD) and headspace solid phase microextraction (HS-SPME) approaches associated to gas chromatography-mass spectrometric (GC-MS) quantification. Using MAHD-GC-MS and HS-SPME-GC-MS techniques, 30 and 28 constituents were screened in the essential oils and volatiles of M. pulegium L. aerial parts with high prevalence of oxygenated monoterpenes and non-terpene hydrocarbons, respectively. Accordingly, in the characterized chemical profiles, carvone was found to constitute about 56.0% of the oil using the former technique (MAHD), whereas oleic acid (20.1%), carvone (17.7%) along with limonene (16.1%) were found as the major constituent components of the volatile profile using the latter approach (SPME). The two methods might be used in combination to obtain a wider set of information about the chemical composition of one specific plant sample.

Bioengineered synthesis of phytochemical-adorned green silver oxide (Ag2O) nanoparticles via Mentha pulegium and Ficus carica extracts with high antioxidant, antibacterial, and antifungal activities.

Silver oxide nanoparticles have various biomedical and pharmaceutical applications. However, conventional nanofabrication of Ag2O is associated with the use of toxic chemicals and organic solvents. To circumvent this hurdle, herein silver oxide quantum dots (Ag2O-QDs) were synthesized quickly (3 min) via the use of ultrasonic irradiation and plant-extract. Additionally, due to ultrasonic irradiation's effect on cell-wall destruction and augmentation of extraction efficiency, ultrasonic was also used in the preparation of Mentha pulegium and Ficus carica extracts (10 min, r.t) as natural eco-friendly reducing/capping agents. The UV-Vis result indicated a broad absorption peak at 400-500 nm. TEM/SEM analysis showed that ultrasound introduced a uniform spherical particle and significantly reduced particle size compared to the conventional heating method (∼ 9 nm vs. ∼ 100 nm). Silver and oxygen elements were found in the bio-synthesized Ag2O by EDS. The FTIR and phenol/flavonoid tests revealed the presence of phenol and flavonoid associated with the nanoparticles. Moreover, nanoparticles exhibited antioxidant/antibacterial/antifungal activities. The MIC and MBC results showed the Ag2O QDs synthesized with M. pulegium extract have the highest antibacterial activity against E. coli (MBC = MIC:15.6 ppm), which were significantly different from uncoated nanoparticles (MBC = MIC:500 ppm). The data reflects the role of phyto-synthesized Ag2O-QDs using ultrasonic-irradiation to develop versatile and green biomedical products.

Molecular Interaction Studies and Phytochemical Characterization of Mentha pulegium L. Constituents with Multiple Biological Utilities as Antioxidant, Antimicrobial, Anticancer and Anti-Hemolytic Agents.

Multiple biological functions of Mentha pulegium extract were evaluated in the current work. Phytochemical components of the M. pulegium extract were detected by Gas Chromatography-Mass Spectrometry (GC-MS) and High-performance liquid chromatography (HPLC). Moreover, M. pulegium extract was estimated for antioxidant potential by 2,2-Diphenyl-1-picryl-hydrazyl-hydrate (DPPH) free radical scavenging, antimicrobial activity by well diffusion, and anticoagulant activity via prothrombin time (PT) and activated partial thromboplastin time (APTT). GC-MS analysis detected compounds including cholesterol margarate, stigmast-5-en-3-ol, 19-nor-4-androstenediol, androstan-17-one, pulegone-1,2-epoxide, isochiapin B, dotriacontane, hexadecanoic acid and neophytadiene. Chrysoeriol (15.36 µg/mL) was followed by kaempferol (11.14 µg/mL) and 7-OH flavone (10.14 µg/mL), catechin (4.11 µg/mL), hisperdin (3.05 µg/mL), and luteolin (2.36 µg/mL) were detected by HPLC as flavonoids, in addition to ferulic (13.19 µg/mL), cinnamic (12.69 µg/mL), caffeic (11.45 µg/mL), pyrogallol (9.36 µg/mL), p-coumaric (5.06 µg/mL) and salicylic (4.17 µg/mL) as phenolics. Antioxidant activity was detected with IC50 18 µg/mL, hemolysis inhibition was recorded as 79.8% at 1000 µg/mL, and PT and APTT were at 21.5 s and 49.5 s, respectively, at 50 µg/mL of M. pulegium extract. The acute toxicity of M. pulegium extract was recorded against PC3 (IC50 97.99 µg/mL) and MCF7 (IC50 80.21 µg/mL). Antimicrobial activity of M. pulegium extract was documented against Bacillus subtilis, Escherichia coli, Pseudomonasaureus, Candida albicans, Pseudomonas aeruginosa, but not against black fungus Mucor circinelloides. Molecular docking was applied using MOE (Molecular Operating Environment) to explain the biological activity of neophytadiene, luteolin, chrysoeriol and kaempferol. These compounds could be suitable for the development of novel pharmacological agents for treatment of cancer and bacterial infections.

Response surface modeling and optimization of the extraction conditions using lactic acid-based deep eutectic solvents as green alternative extraction media for Mentha pulegium.

INTRODUCTION: Most recurrently available organic solvents are toxic and inflammable and pose high risks to human health. Natural deep eutectic solvents (NADESs) have been developed as promising green alternatives. OBJECTIVE: We aimed to extract polyphenolic compounds from Mentha pulegium using lactic acid-based deep eutectic solvents. Extraction parameters were optimized by response surface methodology. MATERIAL AND METHODS: Combined with ultrasound-assisted extraction, three different lactic acid-based deep eutectic solvents were investigated for the extraction of polyphenols. Methanol (80%, v/v) was used for comparison. The optimized influencing factors were: water content in solvent, extraction time, and temperature. The design was adopted including 17 experiments with three center points. RESULTS: All NADESs tested showed an excellent extraction efficacy compared to 80% methanol. Under the optimized conditions, with 45% of water, at 30°C, and for extraction 90 min, the highest extraction yields were recorded using lactic acid:sodium acetate (3:1), achieving 173.35 ± 0.02 mg gallic acid equivalents (GAE)/g dry weight (dw) of polyphenols and 95 ± 0.09% antioxidant activity. After extraction for 90 min at 80°C with 18% of water, we obtained 164.06 ± 0.01 mg GAE/g dw and 94 ± 0.02% antioxidant activity using lactic acid:glucose (5:1). Efficient recovery (64.92 ± 0.01 mg GAE/g dw and 97 ± 0.1% antioxidant activity) was achieved using lactic acid:glycine (3:1) with 31% of water, at 35°C, and extraction for 30 min. CONCLUSION: Our results indicate that with optimized parameters, the proposed natural solvents are excellent alternatives to chemical ones for the extraction of phenolic compounds.

Promising Larvicidal Efficacy of Solid Lipid Nanoparticles Containing Mentha longifolia L., Mentha pulegium L., and Zataria multiflora Boiss. Essential Oils Against the Main Malaria Vector, Anopheles stephensi Liston.

PURPOSE: An attempt was made in the current study to develop a natural mosquito larvicide using nanotechnology. METHODS: Solid lipid nanoparticles (SLNs) containing three essential oils were first prepared using the high-pressure homogenizer. Larvicidal effects of essential oils and the SLNs against Anopheles stephensi were then compared. RESULTS: The size of SLN containing Mentha longifolia, Mentha pulegium, Zataria multiflora essential oil was obtained as 105 ± 7, 210 ± 4, and 137 ± 8 nm. Their zeta potentials were - 7.8, - 4.7, and - 9.7 mV. Besides, their efficacy with LC50 values of 24.79, 5.11, and 9.19 µg/mL was significantly more potent than that of their un-formulated essential oils with LC50 values of 36.2, 27.55, and 33.33 µg/mL. CONCLUSION: SLNs containing M. pulegium with the best efficacy (P < 0.05) could be considered as potent larvicides against other important species of mosquitoes and field trials.

Mentha pulegium L. (Pennyroyal, Lamiaceae) Extracts Impose Abortion or Fetal-Mediated Toxicity in Pregnant Rats; Evidenced by the Modulation of Pregnancy Hormones, MiR-520, MiR-146a, TIMP-1 and MMP-9 Protein Expressions, Inflammatory State, Certain Related Signaling Pathways, and Metabolite Profiling via UPLC-ESI-TOF-MS.

Pregnant women usually turn to natural products to relieve pregnancy-related ailments which might pose health risks. Mentha pulegium L. (MP, Lamiaceae) is a common insect repellent, and the present work validates its abortifacient capacity, targeting morphological anomalies, biological, and behavioral consequences, compared to misoprostol. The study also includes untargeted metabolite profiling of MP extract and fractions thereof viz. methylene chloride (MecH), ethyl acetate (EtOAc), butanol (But), and the remaining liquor (Rem. Aq.) by UPLC-ESI-MS-TOF, to unravel the constituents provoking abortion. Administration of MP extract/fractions, for three days starting from day 15th of gestation, affected fetal development by disrupting the uterine and placental tissues, or even caused pregnancy termination. These effects also entailed biochemical changes where they decreased progesterone and increased estradiol serum levels, modulated placental gene expressions of both MiR-(146a and 520), decreased uterine MMP-9, and up-regulated TIMP-1 protein expression, and empathized inflammatory responses (TNF-α, IL-1ß). In addition, these alterations affected the brain's GFAP, BDNF, and 5-HT content and some of the behavioral parameters escorted by the open field test. All these incidences were also perceived in the misoprostol-treated group. A total of 128 metabolites were identified in the alcoholic extract of MP, including hydroxycinnamates, flavonoid conjugates, quinones, iridoids, and terpenes. MP extract was successful in terminating the pregnancy with minimal behavioral abnormalities and low toxicity margins.

Phytochemistry, anti-tick, repellency and anti-cholinesterase activities of Cupressus sempervirens L. and Mentha pulegium L. combinations against Hyalomma scupense (Acari: Ixodidae).

The adulticidal, larvicidal, and repellent effects of Cupressus sempervirens, Mentha pulegium, and their combination (C. sempervirens and M. pulegium) against Hyalomma scupense were investigated, as well as their putative mode of action. The evaluation on H. scupense was performed with the adult immersion test at concentrations ranging from 2.5-20 mg/mL and the larval packet test from 0.625 to 20 mg/mlL. The same concentrations were used for combined oils. Essential oils were obtained using hydrodistillation and then analyzed using GC-MS. The following major components were identified: α-pinene (56.89 %) and δ-3-Carene (15.61 %) for C. sempervirens, while pulegone (70.50 %) and cis-Menthone (18.26 %) for M. pulegium. At 20 mg/mL concentration, C. sempervirens had (94.40 %) acaricide action and inhibited the egg from hatching, however at 10 mg/mL, it had 100 % larvicidal activity. Adulticidal and larvicidal activities of M. pulegium oil reached to 87.19 % and 81.02 % respectively, at 20 mg/mL concentration. The highest acaricidal property against H. scupense was exhibited by combination oils with minimum LC50 values of 1.76 mg/mL on adultes and 0.60 mg/mL on larvae. The repellent activities were 100, 95.80 and 100 %, at the 20 mg/mL concentration used in 6 h, for C. sempervirens, M. pulegium and mixture oil respectively. Our findings revealed that the mixture of C. sempervirens and M. pulegium, at the ratio 1:1 (v: v) of each oil, showed high synergistic effect (SF > 1) and the use of combination oil (C. sempervirens 50 %: M. pulegium 50 %) is more toxic against H.scupense. Furthermore, the volatile oils examined revealed significant inhibition of H.scupense acetylcholinesterase activity AChE. Based on the findings of the present framework, these essential oils have the potential to be applied as substitute for synthetic acaricides in the management of H.scupense bovine ticks.

Chemical Composition and Synergistic Potential of Mentha pulegium L. and Artemisia herba alba Asso. Essential Oils and Antibiotic against Multi-Drug Resistant Bacteria.

The essential oils were obtained by hydrodistillation from aerial parts of Mentha pulegium L. (M. pulegium L.) and Artemisia herba alba (A. herba alba) Asso. and analyzed by gas chromatography-flame ionization detector chromatograpy (GC-FID) and gaz chromatography-mass spectrometry (GC-MS). The antibacterial activities of the oils were determined by the disk diffusion method and a microdilution broth assay against six bacteria stains. The combinations of these essential oils with antibiotics were evaluated against two multi-drug-resistant bacteria strains: imipenem-resistant Acinetobacter baumannii (IRAB S3310) and methicillin-resistant Staphylococcus aureus (MRSA S19). The chemical analysis of M. pulegium essential oil revealed the presence of pulegone (74.8%) and neoisomenthol (10.0%). A. herba alba essential oil was characterized by camphor (32.0%), α-thujone (13.7%), 1,8-cineole (9.8%), ß-thujone (5.0%), bornéol (3.8%), camphene (3.6%), and p-cymene (2.1%). All strains tested except Pseudomonas aeruginosa were susceptible to these oils. The combinations of essential oils with antibiotics exerted synergism, antagonism, or indifferent effects. The best effect was observed with A. herba alba essential oil in association with cefoxitin (CX) against MRSA S19. However, for IRAB S3310, the strongest synergistic effect was observed with M. pulegium in association with amikacin (AK). This study demonstrated that M. pulegium and A. herba alba essential oils have antibacterial activities which could be potentiated by antibiotics especially in the case of IRAB S3310.

Tunisian Native Mentha pulegium L. Extracts: Phytochemical Composition and Biological Activities.

Mint species (Lamiaceae family) have been used as traditional remedies for the treatment of several diseases. In this work, we aimed to characterize the biological activities of the total phenolic and flavonoid contents of Mentha pulegium L. extracts collected from two different regions of Tunisia. The highest amounts of total phenols (74.45 ± 0.01 mg GAE/g DW), flavonoids (28.87 ± 0.02 mg RE/g DW), and condensed tannins (4.35 ± 0.02 mg CE/g DW) were found in the Bizerte locality. Methanolic leaf extracts were subjected to HPLC-UV analysis in order to identify and quantify the phenolic composition. This technique allowed us to identify seven phenolic compounds: two phenolic acids and five flavonoid compounds, such as eriocitrin, hesperidin, narirutin, luteolin, and isorhoifolin, which were found in both extracts with significant differences between samples collected from the different regions (p < 0.05). Furthermore, our results showed that the methanolic extract from leaves collected from Bizerte had the highest antioxidant activities (DPPH IC50 value of 16.31 µg/mL and 570.08 µmol Fe2+/g, respectively). Both extracts showed high radical-scavenging activity as well as significant antimicrobial activity against eight tested bacteria. The highest antimicrobial activities were observed against Gram-positive bacteria with inhibition zone diameters and MIC values ranging between 19 and 32 mm and 40 and 160 µg/mL, respectively. Interestingly, at 10 µg/mL, the extract had a significant effect on cell proliferation of U87 human glioblastoma cells. These findings open perspectives for the use of Mentha pulegium L. extract in green pharmacy, alternative/complementary medicine, and natural preventive therapies for the development of effective antioxidant, antibacterial, and/or antitumoral drugs.

Mentha pulegium L.: A Plant Underestimated for Its Toxicity to Be Recovered from the Perspective of the Circular Economy.

The aim of the study was to investigate the micromorphology of Mentha pulegium leaves and flowers harvested in three different Sicilian (Italy) areas with peculiar pedo-climatic conditions, and to characterize the phytochemical profile, the phytotoxic activity, and the eco-compatibility of their essential oils (EOs) for potential use as safe bioherbicides. Light microscopy (LM) and scanning electron microscopy (SEM) highlighted that M. pulegium indumentum consists of non-glandular and glandular trichomes of different types. Peltate trichomes of plants from the different sites showed few significant differences in dimension and abundance, but they were characterized by a surprisingly high number of secretory cells both in leaves and flowers. Phytochemical analyses showed that oxygenated monoterpenes were the most abundant class in all the EOs investigated (92.2-97.7%), but two different chemotypes, pulegone/isomenthone and piperitone/isomenthone, were found. The complex of morphological and phytochemical data indicates that soil salinity strongly affects the expression of the toxic metabolite pulegone, rather than the EO yield. Phytotoxicity tests showed a moderate activity of EOs against the selected species as confirmed by α-amylase assay. Moreover, the low toxicity on brine shrimp provided a rationale for the possible use of investigated EOs as eco-friendly herbicides.

Essential oil of Mentha pulegium induces anthelmintic effects and reduces parasite-associated oxidative stress in rodent model.

Following the previous findings reported by the present authors on the anthelmintic effect of hydro-ethanolic extract of Mentha pulegium, the volatile constituents of M. pulegium are now assessed in the present study by exploring its anthelmintic and its antioxidant proprieties using in vitro and in vivo assays. Egg hatch assay (EHA) and adult worm's motility assays (AWMA) were used to assess the in vitro activity against Haemonchus. contortus. The in vivo anthelmintic potential was evaluated in mice infected with Heligmosomoides polygyrus using faecal egg count reduction (FECR) and total worm count reduction (TWCR). M. pulegium EO demonstrated 100% inhibition in the EHA at 200 µg/mL (IC50 = 56.36 µg/mL). In the AWM assay, EO achieved total worms paralysis 6 h after treatment exposure. This nematicidal effect was associated to morphological damages observed in the cuticular's worm using environmental scanning electron microscopy (ESEM). At 400 mg/kg, M. pulegium oil showed 75.66% of FECR and 80.23% of TWCR. The antioxidant potential of this plant was also monitored by several in vitro assays: total antioxidant capacity was 205.22 mg GAE/g DW, DPPH quenching effect was IC50 = 140 µg/mL, ABTS activity IC50 = 155 µg/mL and FRAP effect of 660 µg/mL. Regarding the in vivo assay, M. pulegium EO demonstrated a protective effect against oxidative stress by increasing the activity of the endogenous antioxidants (SOD, CAT and GPx) during H. polygyrus infection.