Chemical Profiling, Insecticidal, and Phytotoxic Effect of Essential Oils from Leaves and Inflorescence of Moroccan Chenopodium ambrosioides (L.).

Chenopodium ambrosioides (L.) is a medicinal and aromatic plant widely used in the Moroccan traditional medicine for its important pharmacological properties. In order to contribute to the valorization of this plant, the current study aimed at evaluating, for the first time, the variation in the yield, chemical composition, and insecticidal and phytotoxic activities of C. ambrosioides essential oil (CAEO) isolated from leaves and inflorescences. The results obtained showed that the CAEO yields vary significantly according to the distilled plant part, with 0.75 ± 0.15% for the leaves and 1.2 ± 0.34% for the inflorescences. CAEO profiling showed quantitative more than qualitative differences. Leaf CAEO was rich in δ-3-carene (61.51%), followed by p-cymene (14.67%) and 1,2:3,4-diepoxy-p-menthane (6.19%). However, inflorescence CAEO was dominated by the same compounds but with variable levels (δ-3-Carene: 44.29%; 1,2:3,4-diepoxy-p-menthane: 19.46%; and p-cymene: 17.85%). The CAEOs from the leaves and inflorescences showed a very interesting inhibiting effect on the germination and growth of the three species used, namely Medicago sativa, Linum rusitatissimum, and Raphanus sativus. However, there was no significant difference between these CAEOs. Similarly, for the insecticidal activity, CAEOs exhibited an important and similar toxicity against Tribolium confusum adults with LD50 of 4.30 and 4.46 µL/L of air and LD90 of 6.51 and 9.62 µL/L air for toxicity by fumigation, while values for contact toxicity on filter paper were 0.04 and 0.05 µL/cm2 for LD50 and 0.08 and 0.09 µL/cm2 for LD90.

New insight into antimicrobial activities of Linaria ventricosa essential oil and its synergetic effect with conventional antibiotics.

The purpose of the present study was to determine for the first time the volatile constituents, the antioxidant and antimicrobial activities of the essential oil (EO) of the endemic Moroccan Linaria ventricosa, alone or in combination with four known antibiotics. The major constituents were 2-methoxy-4-vinylphenol (17.4%), α-terpinene (13.64%) and 3,5-dimethylphenyl isocyanate (12.21%). The EO had moderate antioxidant potency, as measured by DPPH free radical scavenging (1.233 ± 0.031 mg/mL), ferric reducing antioxidant power assay (0.373 ± 0.019 mg/mL) and ß-carotene/linoleic acid (0.922 ± 0.026 mg/mL). EO showed microbicidal activity against all microorganisms tested. The highest effectiveness was recorded against Candida albicans (IZ = 24 mm, MIC = 4.87 mg/mL and MMC = 9.75 mg/mL) and Candida glabrata (IZ = 22 mm, MIC = MMC = 4.87 mg/mL). Gram negative bacteria were the most resistant (MIC = MMC = 39 mg/mL). The combination of EO at sub-inhibitory concentrations with antibiotics showed a significant decrease in their individual MICs from 2 to 128 fold, being the best for ciprofloxacin and fluconazole against E. coli and C. albicans and C. glabrata, respectively.

Seaweed Essential Oils as a New Source of Bioactive Compounds for Cyanobacteria Growth Control: Innovative Ecological Biocontrol Approach.

The application of natural compounds extracted from seaweeds is a promising eco-friendly alternative solution for harmful algae control in aquatic ecosystems. In the present study, the anti-cyanobacterial activity of three Moroccan marine macroalgae essential oils (EOs) was tested and evaluated on unicellular Microcystis aeruginosa cyanobacterium. Additionally, the possible anti-cyanobacterial response mechanisms were investigated by analyzing the antioxidant enzyme activities of M. aeruginosa cells. The results of EOs GC-MS analyses revealed a complex chemical composition, allowing the identification of 91 constituents. Palmitic acid, palmitoleic acid, and eicosapentaenoic acid were the most predominant compounds in Cystoseira tamariscifolia, Sargassum muticum, and Ulva lactuca EOs, respectively. The highest anti-cyanobacterial activity was recorded for Cystoseira tamariscifolia EO (ZI = 46.33 mm, MIC = 7.81 µg mL-1, and MBC = 15.62 µg mL-1). The growth, chlorophyll-a and protein content of the tested cyanobacteria were significantly reduced by C. tamariscifolia EO at both used concentrations (inhibition rate >67% during the 6 days test period in liquid media). Furthermore, oxidative stress caused by C. tamariscifolia EO on cyanobacterium cells showed an increase of the activities of superoxide dismutase (SOD) and catalase (CAT), and malondialdehyde (MDA) concentration was significantly elevated after 2 days of exposure. Overall, these experimental findings can open a promising new natural pathway based on the use of seaweed essential oils to the fight against potent toxic harmful cyanobacterial blooms (HCBs).

A Comparative Study of the in Vitro Antimicrobial and Synergistic Effect of Essential Oils from Laurus nobilis L. and Prunus armeniaca L. from Morocco with Antimicrobial Drugs: New Approach for Health Promoting Products.

Laurus nobilis L. (laurel, Lauraceae) and Prunus armeniaca L. (apricot, Rosaceae) are important industrial crops and display significant biological properties, including antimicrobial activity. In this work, essential oils (EOs) prepared from the leaves of both species from Morocco were evaluated for the first time for possible synergistic in vitro antibacterial and antifungal effects with some conventional antimicrobial drugs, namely fluconazole, ciprofloxacin and vancomycin. Samples were further evaluated for chemical composition by gas chromatography-mass spectrometry (GC-MS). The main volatile compounds detected in L. nobilis were eucalyptol (40.85%), α-terpinyl acetate (12.64%) and methyl eugenol (8.72%), while P. armeniaca was dominated essentially by (Z)-phytol (27.18%), pentacosane (15.11%), nonacosane (8.76%) and benzaldehyde (7.25%). Regarding antimicrobial activity, both EOs inhibited significantly all the microorganisms tested. The EO from L. nobilis had the highest activity, with minimal inhibitory concentrations (MICs) ranging from 1.39 to 22.2 mg/mL for bacteria and between 2.77 and 5.55 mg/mL for yeasts. Conversely, the combination of the studied EOs with ciprofloxacin, vancomycin and fluconazol resulted in a noteworthy decrease in their individual MICs. In fact, of the 32 interactions tested, 23 (71.87%) demonstrated total synergism and 9 (28.12%) a partial synergistic interaction. The EO from L. nobilis exhibited the highest synergistic effect with all the antibiotics used, with fractional inhibitory concentration (FIC) index values in the range of 0.266 to 0.75 for bacteria, and between 0.258 and 0.266 for yeast. The synergistic interaction between the studied EOs and standard antibiotics may constitute promising anti-infective agents useful for treating diseases induced by antibiotic-resistant pathogens.

Endophytic actinobacteria of medicinal plant Aloe vera: Isolation, antimicrobial, antioxidant, cytotoxicity assays and taxonomic study

Objective: To explore the new sources of novel bioactive compounds having pharmaceutical and agricultural interest and to search the endophytic actinobacteria from medicinal plants. Methods: NAF-1 an endophyte actinobacteria was isolated from leaves of medicinal plant Aloe vera collected in Marrakesh, Morocco using Bennett agar as selective medium. NAF-1 was tested for its antimicrobial activity against five pathogenic bacteria such asStaphylococcus aureus PIC 53156,Micrococcus luteus ATCC381,Bacillus subtilis ATCC 14579,Pseudomonas aeruginosa DSM 50090 and Escherichia coli ATCC 8739 and four human clinic fungi belonging to the Candida,Aspergillus and Microsporum genera. Several antioxidant activities were studied such as DPPH free radical scavenging,β-carotene and linoleic acid and reducing power assays. The total of phenol and flavonoid was also calculated. Using Artemia salina shrimp assay, the cytotoxicity of NAF-1 crude extract was determined.Results: The results revealed that the actinobacteria showed a high activity (≥20 mm) against only Gram positive bacteria but it had a moderate activity (between 13 and 15 mm) against Human clinic fungi. The isolate also exhibited a LD50 of 14.20 μg/mL in the cytotoxicity assay. The result showed that the crude extract presented an interesting free radical-scavenging activity with IC50 value of (5.58 ± 0.26) μg/mL and a high value of phenolic and flavonoid compounds with (15.41 ± 0.18) μg GAE/mg extract and (11.41± 0.06) μg QE/mg extract respectively. Moreover, the taxonomic position of our endophyte actinobacteria using the morphological and physiological criteria and using16SrRNA gene sequence (polyphasic approach) showed that the NAF-1 isolate was similar to Streptomyces hydrogenans which was never described as an endophyte actinobacteria. Conclusions: This isolated strain appears promising resources of bioactive agents and can be exploited to produce therapeutic agents active against pathogenic disease.

Chemical Characterization and Biological Activities of Essential Oil Obtained from Mint Timija Cultivated under Mineral and Biological Fertilizers.

Cultivation of mint timija (Mentha suaveolens subsp. timija (Briq.) Harley) constitutes a promising solution to the conservation and sustainable utilization of this Moroccan endemic and threatened species. Optimized agronomic practices require mineral and/or biological fertilizer applications. The aim of this study was to determine the effects of application of a complete (N, P, and K) mineral fertilizer and vesicular arbuscular mycorrhizae (VAM) inoculation on the composition, antioxidant, and insecticidal properties of mint timija essential oils (EOs). The GC-MS analyses identified 27 components representing more than 99.9% of the total oils. Menthone (40.7-49.3%), pulegone (31.3-36.5%), and isomenthone (2.5-4.4%) were found to be the main constituents. Cultivation of mint timija with mineral fertilizer and VAM inoculation induced an increase in menthone content and a parallel decrease of pulegone. Both treatments enhanced the antioxidant activity of the investigated EOs in all assays (IC50 ranged from 2.34 ± 0.03 mg/mL to 6.82 ± 0.25 mg/mL), while no significant difference in the toxicities of these oils against Tribolium confusum du Val. has been observed. Overall, we conclude that cultivation using complete mineral fertilizer and VAM inoculation could be useful in modulating the chemical composition and enhancing the antioxidant activity of the EO of this endemic Moroccan species.

Chemical characterization and insecticidal properties of essential oils from different wild populations of Mentha suaveolens subsp. timija (Briq.) Harley from Morocco.

The present study is the first investigation of the volatile-oil variability and insecticidal properties of the endemic Moroccan mint Mentha suaveolens subsp. timija (mint timija). The yield of essential oils (EOs) obtained from different wild mint timija populations ranged from 0.20±0.02 to 1.17±0.25% (v/w). GC/MS Analysis revealed the presence of 44 oil constituents, comprising 97.3-99.9% of the total oil compositions. The main constituents were found to be menthone (1.2-62.6%), pulegone (0.8-26.6%), cis-piperitone epoxide (2.9-25.5%), piperitone (0.3-35.5%), trans-piperitone epoxide (8.1-15.7%), piperitenone (0.2-9.6%), piperitenone oxide (0.5-28.6%), (E)-caryophyllene (1.5-11.0%), germacrene D (1.0-15.7%), isomenthone (0.3-7.7%), and borneol (0.2-7.3%). Hierarchical-cluster analysis allowed the classification of the EOs of the different mint timija populations into four main groups according to the contents of their major components. This variability within the species showed to be linked to the altitude variation of the mint timija growing sites. The results of the insecticidal tests showed that all samples exhibited interesting activity against adults of Tribolium castaneum, but with different degrees. The highest toxicity was observed for the EOs belonging to Group IV, which were rich in menthone and pulegone, with LC50 and LC90 values of 19.0-23.4 and 54.9-58.0 µl/l air in the fumigation assay and LC50 and LC90 values of 0.17-0.18 and 0.40-0.52 µl/cm(2) in the contact assay.

Comparative evaluation of antioxidant and insecticidal properties of essential oils from five Moroccan aromatic herbs.

This study describes the antioxidant and insecticidal activities of essential oils (EOs) of Mentha suaveolens subsp. timija, Thymus satureioides, Achillea ageratum, Cotula cinerea and Salvia officinalis widely used in Morocco as flavorings, food additives and preservatives. Sixty seven components were identified accounting for more than 95.0 % of the total oils. M. suaveolens subsp. timija oil had as main components menthone and pulegone. A. ageratum oil was particularly rich in artemisyl acetate and yomogi alcohol. The essential oil of T. satureioides was characterized by high contents of carvacrol and borneol. C. cinerea oil contained trans-thujone and cis-verbenyl acetate as major constituents, whereas S. officinalis oil was characterized by trans-thujone and camphor. Antioxidant activities were examined by means of 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), reducing power, ß-carotene/linoleic acid bleaching and ABTS radical tests. In all assays, the highest antioxidant potency was observed in T. satureioides EO with IC50 values ranging from 0.15 ± 0.36 µg mL(-1) to 0.23 ± 0.67 µg mL(-1) across the four assays. The in vitro evaluation of the insecticidal activity showed that M. suaveolens subsp. timija EO present the highest insecticidal efficiency against adults of Tribolium castaneum with LD50 and LD90 values of 0.17 µL cm(-2) and 0.26 µL cm(-2), respectively and LT50, LT90 values ranged from 44.19 h to 2.98 h and 98.14 h to 6.02 h, respectively. Our data support the possible use of T. satureioides oil as potential antioxidant agent, while M. suaveolens subsp. timija oil can be developed as a new natural bio-insecticide.

Insecticidal properties and chemical composition of essential oils of some aromatic herbs from Morocco.

The essential oils (EOs) of Artemisia herba alba, Rutachalepensis and Satureja calamintha aerial parts were analysed by GC/MS and the EOs were tested for their toxicity against two flour beetles, Tribolium castaneum and Tribolium confusum. EO composition showed that the major compounds were pulegone, menthone and menthol for S. calamintha, camphor and ß-thujone for A. herba alba and 2-undecanone for R. chalepensis. Contact bioassay showed that all EOs were toxic to adults of T. castaneum and T. confusum. Higher toxicity was found with the EOs from S. calamintha and R. chalepensis (LD50 of 0.09-0.13 µL/cm(2) and LD90 of 0.17-0.29 µL/cm(2)). In fumigant toxicity test, A. herba alba and S. calamintha EOs produced insecticidal activity with S. calamintha EO being more toxic with LD50 and LD90 values of 10.5 and 19.1 µL/L air, respectively, for T. confusum against 7.8 and 17.4 µL/L air, respectively, for T. castaneum.