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.

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.

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.

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.

Towards Bioprospection of Commercial Materials of Mentha spicata L. Using a Combined Strategy of Metabolomics and Biological Activity Analyses.

Spearmint (Mentha spicata L.) has been widely studied for its diversity of compounds for product generation. However, studies describing the chemical and biological characteristics of commercial spearmint materials from different origins are scarce. For this reason, this research aimed to bioprospecting spearmint from three origins: Colombia (Col), Mexico (Mex), and Egypt (Eg). We performed a biological activity analysis, such as FRAP, DPPH, and ABTS, inhibition potential of S. pyogenes, K. pneumoniae, E. coli, P. aeuroginosa, S. aureus, S aureus Methicillin-Resistant, and E. faecalis. Furthermore, we performed chemical assays, such as total polyphenol and rosmarinic acid, and untargeted metabolomics via HPLC-MS/MS. Finally, we developed a causality analysis to integrate biological activities with chemical analyses. We found significant differences between the samples for the total polyphenol and rosmarinic acid contents, FRAP, and inhibition analyses for Methicillin-Resistant S. aureus and E. faecalis. Also, clear metabolic differentiation was observed among the three commercial materials evaluated. These results allow us to propose data-driven uses for the three spearmint materials available in current markets.

Spearmint (Mentha spicata L.) Phytochemical Profile: Impact of Pre/Post-Harvest Processing and Extractive Recovery.

The purpose of this study was to chemically compare samples of Mentha spicata (marketing byproducts, production byproducts, and export material), cultivated in the open field and under greenhouse, using an integrated approach by HPLC/DAD and GC/MS analysis. The presence of phenolic compounds was higher in the marketing byproducts cultivated in the open field. Marketing byproducts also had the highest amount of carvone. For this reason, this byproduct was selected as a candidate for the development of natural ingredients. With the best selected material, the optimization of simultaneous high-intensity ultrasound-assisted extraction processes was proposed for the recovery of the compounds of interest. This extraction was defined by Peleg's equation and polynomial regression analysis. Modeling showed that the factors amplitude, time, and solvent were found to be significant in the recovery process (p < 0.005). The maximum amount of compounds was obtained using 90% amplitude for 5 min and ethanol/water mixture (80:20) for extraction to simultaneously obtain phenolic and terpenoid compounds. This system obtained the highest amount of monoterpenoid and sesquiterpenoid compounds from the essential oil of M. spicata (64.93% vs. 84.55%). Thus, with an efficient and eco-friendly method, it was possible to optimize the extraction of compounds in M. spicata as a starting point for the use of its byproducts.

Bioactive properties of the aromatic molecules of spearmint (Mentha spicata L.) essential oil: a review.

Spearmint belongs to the genus Mentha in the family Labiatae (Lamiaceae), which is cultivated worldwide for its remarkable aroma and commercial value. The aromatic molecules of spearmint essential oil, including carvone, carveol, dihydrocarvone, dihydrocarveol and dihydrocarvyl acetate, have been widely used in the flavors and fragrances industry. Besides their traditional use, these aromatic molecules have attracted great interest in other application fields (e.g., medicine, agriculture, food, and beverages) especially due to their antimicrobial, antioxidant, insecticidal, antitumor, anti-inflammatory and antidiabetic activities. This review presents the sources, properties, synthesis and application of spearmint aromatic molecules. Furthermore, this review focuses on the biological properties so far described for these compounds, their therapeutic effect on some diseases, and future directions of research. This review will, therefore, contribute to the rational and economic exploration of spearmint aromatic molecules as natural and safe alternative therapeutics.

The traditional uses, phytochemistry and pharmacology of spearmint (Mentha spicata L.): A review.

ETHNOPHARMACOLOGY RELEVANCE: Mentha spicata L. (Lamiaceae), commonly called Spearmint, is wildly cultivated worldwide for its remarkable aroma and commercial value. In addition to traditional foods flavouring agent, M. spicata is well known for its traditional medicinal uses, particularly for the treatment of cold, cough, asthma, fever, obesity, jaundice and digestive problems. AIM OF THE REVIEW: This review aims to critically appraise scientific literature regarding the traditional uses, bioactive chemical constituents and pharmacological activities of M. spicata. MATERIALS AND METHODS: A review of the literature information on M. spicata was searched from scientific electronic search databases (Google Scholar, PubMed, Web of Science, ACS, Science Direct, Taylor and Francis, Wiley, Springer and SCOPUS. Structures for secondary metabolites were confirmed using PubChem and ChemSpider. RESULTS: The studies conducted on either crude extracts, essential oil or isolated pure compounds from M. spicata had reported a varied range of biological effects including antibacterial, antifungal, antioxidant, hepatoprotective, antidiabetic, cytotoxic, anti-inflammatory, larvicidal activity, antigenotoxic potential and antiandrogenic activities. Phytochemical analysis of various parts of M. spicata revealed 35 chemical constituents, belonging to phenolic acids, flavonoids and lignans. CONCLUSION: The review finding indicates that the pharmacological properties of M. spicata supported its traditional uses. The essential oils and extracts showed remarkable antimicrobial, antioxidant, anticancer, anti-inflammatory and hepatoprotective activities. However, more studies, especially in vivo experiments and clinical trials of the human to evaluate cellular and molecular mechanisms based pharmacological, bioactive effectiveness and safety investigation, should be undertaken in the future to provide stronger scientific proof for their traditional medicinal properties.

Antifungal activity of essential oil from Mentha spicata L. and Mentha pulegium L. growing wild in Sardinia island (Italy).

This study aims to evaluate the antifungal activity of Mentha spicata L. and Mentha pulegium L. from Sardinia and to assess their efficacy on virulence factors for Candida albicans, particularly on the inhibition of the germ tube formation. The major compounds of the essential oils were carvone (62.9%) for M. spicata and pulegone (86.2%) for M. pulegium. The essential oil from M. spicata showed a more preeminent effect against Cryptococcus neoformans and the dermatophytes Trichophyton rubrum and T. verrucosum (0.32 µL/mL). Both oils were very effective in inhibiting C. albicans germ tube formation, at doses well below their MIC (0.16 µL/mL).

Effects of supplementing hen diet with Lavandula angustifolia and/or Mentha spicata essential oils on production performance, egg quality and blood variables of laying hens.

BACKGROUND: Organic products of animals are getting more accepted by consumers. Using herbal additives may lead to more health animal products. In this research it is hypothesized that Lavandula angustifolia and/or Mentha spicata essential oils would be helpful to enhance production performance in laying hens. OBJECTIVES: This experiment was conducted to evaluate the effects of Lavandula angustifolia and Mentha spicata essential oils on performance, egg traits and blood variables in laying hens. METHODS: 144 Lohmann LSL-Lite laying hens from 42 until 56 weeks of age were used in a completely randomized design in four treatments and six replicates (six birds per replicate). The treatments consisted of: (a) control group (basal diet), (b) basal diet supplemented with 250 mg/kg diet lavender essential oil (LEO), (c) basal diet supplemented with 250 mg/kg diet mint essential oil (MEO), and (d) basal diet supplemented with both LEO and MEO. RESULTS: Using LEO and/or MEO did not affect body weight changes, feed intake, egg weight, egg index, yolk index, Haugh unit, egg shell weight and egg shell thickness. Feeding LEO, individually or in combination with MEO, did not affect FCR compared with the control group (p < .05), however, feeding MEO individually increased feed conversation ratio (FCR) compared to LEO and the control group during 42-56 weeks (p < .05), as well as decreasing egg mass compared to LEO (p < .05). Feeding LEO increased egg production compared to MEO and combination of MEO and LEO (p < .05). CONCLUSIONS: In conclusion, dietary supplemental MEO (250 mg/kg) may increase FCR, and LEO (250 mg/kg) is more effective than MEO (250 mg/kg) for egg production and egg mass purposes; besides MEO (250 mg/kg) negatively affected FCR compared with the control group. In addition, no specific beneficial effect of dietary supplemental MEO and/or LEO on the other measured variables was detected.

Spearmint Extract Containing Rosmarinic Acid Suppresses Amyloid Fibril Formation of Proteins Associated with Dementia.

Neurological dementias such as Alzheimer's disease and Lewy body dementia are thought to be caused in part by the formation and deposition of characteristic insoluble fibrils of polypeptides such as amyloid beta (Aß), Tau, and/or α-synuclein (αSyn). In this context, it is critical to suppress and remove such aggregates in order to prevent and/or delay the progression of dementia in these ailments. In this report, we investigated the effects of spearmint extract (SME) and rosmarinic acid (RA; the major component of SME) on the amyloid fibril formation reactions of αSyn, Aß, and Tau proteins in vitro. SME or RA was added to soluble samples of each protein and the formation of fibrils was monitored by thioflavin T (ThioT) binding assays and transmission electron microscopy (TEM). We also evaluated whether preformed amyloid fibrils could be dissolved by the addition of RA. Our results reveal for the first time that SME and RA both suppress amyloid fibril formation, and that RA could disassemble preformed fibrils of αSyn, Aß, and Tau into non-toxic species. Our results suggest that SME and RA may potentially suppress amyloid fibrils implicated in the progression of Alzheimer's disease and Lewy body dementia in vivo, as well.

Metabolic Engineering of the Native Monoterpene Pathway in Spearmint for Production of Heterologous Monoterpenes Reveals Complex Metabolism and Pathway Interactions.

Spearmint produces and stores large amounts of monoterpenes, mainly limonene and carvone, in glandular trichomes and is the major natural source of these compounds. Towards producing heterologous monoterpenes in spearmint, we first reduced the flux into the native limonene pathway by knocking down the expression of limonene synthase (MsLS) by RNAi method. The MsLS RNAi lines exhibited a huge reduction in the synthesis of limonene and carvone. Detailed GC-MS and LC-MS analysis revealed that MsLS RNAi plants also showed an increase in sesquiterpene, phytosterols, fatty acids, flavonoids, and phenolic metabolites, suggesting an interaction between the MEP, MVA shikimate and fatty acid pathways in spearmint. Three different heterologous monoterpene synthases namely, linalool synthase and myrcene synthase from Picea abies and geraniol synthase from Cananga odorata were cloned and introduced independently into the MsLS RNAi mutant background. The expression of these heterologous terpene synthases resulted mainly in production of monoterpene derivatives. Of all the introduced monoterpenes geraniol showed the maximum number of derivatives. Our results provide new insights into MEP pathway interactions and regulation and reveals the existence of mechanisms for complex metabolism of monoterpenes in spearmint.

Investigation on the Epoxidation of Piperitenone, and Structure-activity Relationships of Piperitenone Oxide for Differentiation-inducing Activity.

Piperitenone oxide, a major chemical constituent of the essential oil of spearmint, Mentha spicata, induces differentiation in human colon cancer RCM-1 cells. In this study, piperitenone oxide and trans-piperitenone dioxide were prepared as racemic forms by epoxidation of piperitenone. The relative configuration between two epoxides in piperitenone dioxide was determined to be trans by 1H NMR analysis and nuclear Overhauser effect spectroscopy (NOESY) in conjunction with density functional theory (DFT) calculations. Optical resolution of (±)-piperitenone oxide by high-performance liquid chromatography (HPLC) using a chiral stationary phase (CSP) afforded both enantiomers with over 98% enantiomeric excess (ee). Evaluation of the differentiation-inducing activity of the synthetic compounds revealed that the epoxide at C-1 and C-6 in piperitenone oxide is important for the activity, and (+)-piperitenone oxide has stronger activity than (-)-piperitenone oxide. The results obtained in this study provide new information on the application of piperitenone oxide and spearmint for differentiation-inducing therapy. Furthermore, natural piperitenone oxide was isolated from M. spicata. The enantiomeric excess of the isolated natural piperitenone oxide was 66% ee. Epoxidation of piperitenone with hydrogen peroxide proceeded in a phosphate buffer under weak basic conditions to give (±)-piperitenone oxide. These results suggest that the nonenzymatic epoxidation of piperitenone, which causes a decrease in the enantiomeric excess of natural piperitenone oxide, is accompanied by an enzymatic epoxidation in the biosynthesis of piperitenone oxide.

Effect of essential oils of Mentha spicata L. and Melaleuca alternifolia Cheel on the midgut of Podisus nigrispinus (Dallas) (Hemiptera: Pentatomidae).

Agricultural pest control is a popular research topic, and essential oils are widely studied because they represent a promising alternative to synthetic insecticides. However, despite the increase in studies on pests, little work has been done on pesticide contamination of the predators feeding on insecticide-affected prey. Therefore, the objective of this study was to evaluate the effects of Lethal Dose 50 (LD50) from the essential oils of Mentha spicata L. and Melaleuca alternifolia Cheel on the histology, including histochemistry (from protein and carbohydrate analysis) and immunohistochemistry (from the evaluation of cellular apoptosis), of the midgut of fifth instar nymphs of Podisus nigrispinus (stinkbug) (Dallas). The periods of analysis were 12, 24, and 48 h after ingestion of Alabama argillacea Hübner caterpillars treated with the respective oils. The oil from M. spicata did not cause histological alterations or apoptosis in the insect. However, there was a reduction in the level of carbohydrates within the 48-h period. After 24 h, the oil of M. alternifolia caused an elongation of digestive cells and, after 48 h, cell lysis with the release of material into the lumen, suggesting tissue necrosis. The immunohistochemical study revealed no apoptotic process. There was a reduction in the neutral carbohydrate levels in the 24- and 48-h periods and in the number of regenerative cells, when compared to the control, after the period of 48 h. These results demonstrate that M. spicata oil has potential for use in cotton fields because it does not affect the vital characteristics of P. nigrispinus. However, the essential oil of M. alternifolia is not suitable for use as a pesticide because it is extremely toxic to predators.

A combination of spearmint and flaxseed extract improved endocrine and histomorphology of ovary in experimental PCOS.

BACKGROUND: Polycystic ovary syndrome (PCOS) is a complex reproduction and endocrine disorder of women in the reproductive age. Spearmint (Mentha spicata L.) has anti-androgenic activity and flaxseed (Linum usitatissimum L.) contains phytoestrogen and was reported to improve PCOS conditions. This study aimed to evaluate PCOS conditions following administration of a mixture of these two plants. METHODS: Twenty-four rats with regular cycles were randomly divided into four groups as control (C) and treatment-control (TC) received a combination of spearmint extract (SE) + flaxseed extract (FE). PCOS was induced in PCOS and treatment (T) groups by a single intramuscular injection of estradiol valerate. The treatment group received a combination of SE and FE for 30 days, 7 weeks after injection of estradiol valerate. Estrous cycles were monitored for 10 days and in the last day animals were sacrificed, ovaries were collected for the histomorphometric study and the serum levels of progesterone, testosterone, estradiol, and dehydroepiandrosterone (DHEA) were measured. RESULT: Significant rise in progesterone and a decrease in testosterone and estradiol with no significant change of DHEA in the T group, was observed in comparison with the PCOS group (P < 0.05). No significant difference noticed between T and control groups (C &CT) regarding evaluated hormones. A significant increase in primary, pre-antral and antral follicles noticed in the T group compared to the PCOS group. The number of cystic follicles decreased in the T group compared with the PCOS group. Granulosa layer thickness increased while the thickness of theca decreased significantly in the T group compared to the PCOS group (P < 0.05). No significant endocrine or histological differences noticed between C and TC groups. CONCLUSION: A combination of flaxseed and spearmint extract improved the endocrine profile and the histomorphometric features of the ovary in the T group compared to the PCOS group.

In vitro activity of essential oils against adult and immature stages of Ctenocephalides felis felis.

Essential oils (EOs) are considered a new class of ecological products aimed at the control of insects for industrial and domestic use; however, there still is a lack of studies involving the control of fleas. Ctenocephalides felis felis, the most observed parasite in dogs and cats, is associated with several diseases. The aim of this study was to evaluate the in vitro activity, the establishment of LC50 and toxicity of EOs from Alpinia zerumbet (Pers.) B. L. Burtt & R. M. Sm, Cinnamomum spp., Laurus nobilis L., Mentha spicata L., Ocimum gratissimum L. and Cymbopogon nardus (L.) Rendle against immature stages and adults of C. felis felis. Bioassay results suggest that the method of evaluation was able to perform a pre-screening of the activity of several EOs, including the discriminatory evaluation of flea stages by their LC50. Ocimum gratissimum EO was the most effective in the in vitro assays against all flea stages, presenting adulticide (LC50 = 5.85 µg cm-2), ovicidal (LC50 = 1.79 µg cm-2) and larvicidal (LC50 = 1.21 µg cm-2) mortality at low doses. It also presented an excellent profile in a toxicological eukaryotic model. These findings may support studies involving the development of non-toxic products for the control of fleas in dogs and cats.

Comparative laboratory efficacy of novel botanical extracts against Tribolium castaneum.

BACKGROUND: Citrus sinensis, Allium sativum, Ocimum basilicum, Mentha spicata, Curcuma longa, Carica papaya, Andrographis paniculata, Azadirachta indica, Cymbopogon citratus and Acorus calamus were compared in the form of extracts of chloroform, hexane and aqueous media for their effectiveness in repelling Tribolium castaneum. Filter paper of 9 cm in diameter was cut into halves. Using a micro-pipette, each extract was applied to one half of the filter paper. The volume applied was kept constant at 0.6 mL per 30 cm2 . The filter paper was dried and reattached leaving a 0.5 cm gap between, in a Petri-plate. Ten adults of T. castaneum were introduced and the repellency was observed after 30, 60 and 180 min. RESULTS: The aqueous extract of C. papaya exhibited the highest repellency (82-97%) followed by that of garlic bulbs (aqueous extract) with 86.67% repellency. Further, gunny bags were sprayed separately using bottle sprayers with the two most effective extracts at a constant volume of 42 mL per 575 cm2 . These treated gunny bags were compared with malathion-treated gunny bags in the insect management unit. The observations were taken after 2, 8 and 24 h. Papaya leaf extract-treated gunny bags showed relative repellent activity similar to that of malathion-treated gunny bags after 24 h of treatment. Thus, gas chromatography-mass spectrometry analysis of aqueous extract of papaya leaves was performed. CONCLUSIONS: It can be concluded that extract of papaya leaves containing 2-methoxy-4-vinylphenol (8.47% peak area) can be used as a repellent biopesticide. © 2019 Society of Chemical Industry.