In Vitro Methods for Assessing the Antibacterial and Antibiofilm Properties of Essential Oils as Potential Root Canal Irrigants-A Simplified Description of the Technical Steps.

BACKGROUND: Essential oils have gained in significance due to their various biological activities, and there is a growing demand for them in many industries. The present article focuses on the technical steps for an in vitro evaluation of the antibacterial and antibiofilm activities of essential oils for potential use as root canal irrigant in dentistry. METHODS: The bioactivities of the essential oil were investigated through in vitro assays. The gram-positive bacterium Enterococcus faecalis was used as a micro-organism model. The antibacterial activity of the essential oil was assessed using the microdilution method, and resazurin staining to determine the minimal inhibition concentrations (MICs) and the minimal bactericidal concentrations (MBCs). The antibiofilm effect was evaluated spectrophotometrically at 570 nm using the microplate cultivation technique and crystal violet staining. CONCLUSIONS: This article features a detailed in vitro protocol to facilitate the preparation of the essential oil samples, the bacterial suspension, and the methods used for assessment of the antibiofilm and antibacterial activities of the essential oil. The advantages of these approaches are presented in relation to the limits linked to the choice of the bacteria and the essential oil.

Target Enzymes of Origanum majorana and Rosmarinus officinalis Essential Oils in Black Cutworm (Agrotis ipsilon): In Vitro and In Silico Studies.

In this study, in vitro and in silico approaches were employed to assess the toxicity of marjoram (Origanum majorana) and rosemary (Rosmarinus officinalis) essential oils (EOs) to A. ipsilon larvae. The study determined the activities of ATPases in the larvae after treatment with the LC20 and LC70 of each EO. α-esterase and glutathione-S-transferase (GST) activities were also determined after treatment with LC10 and LC30 of each EO. Furthermore, molecular docking was employed to determine the binding affinity of terpinene-4-ol and α-pinene, the major constituents of O. majorana, and R. officinalis EOs, respectively, compared to the co-crystallized ligand of α-esterase, diethyl hydrogen phosphate (DPF). Toxicity assays revealed that O. majorana EO was more toxic than R. officinalis EO to the A. ipsilon larvae at 96 h post-treatment. However, the LC20 and LC70 of the latter significantly inhibited the activity of the Na+-K+ pump at almost all intervals. The same concentrations significantly inhibited the Mg2+/Ca2+-ATPase and Ca2+ pump at 96 h post-treatment. In contrast, O. majorana EO showed a variable effect on the Na+-K+ pump across different time intervals. On the other hand, LC10 and LC30 of both EOs showed varied effects on α-esterase and GST over time. Molecular docking revealed energy scores of -4.51 and -4.29 kcal/mol for terpinene-4-ol and α-pinene, respectively, compared to a score of -4.67 for PDF. Our study demonstrated the toxicity of the tested EOs to A. ipsilon, suggesting their potential efficacy as insecticides.

The Effects of Natural Insecticides on the Green Peach Aphid Myzus persicae (Sulzer) and Its Natural Enemies Propylea quatuordecimpunctata (L.) and Aphidius colemani Viereck.

Botanical insecticides and soaps are frequently proposed as environmentally safer alternatives to synthetic insecticides. However, the efficacy and selectivity of these products are often only partially supported by empirical evidence. Here, we tested the effectiveness of five botanical insecticides, belonging to different categories, on the green peach aphid Myzus persicae (Sulzer) and their selectivity towards two natural enemies, the ladybird beetle Propylea quatuordecimpunctata (L.) and the parasitoid Aphidius colemani (Dalman). White thyme essential oil (EO), sweet orange EO, crude garlic extract and Marseille soap were tested and compared with a pyrethrin-based commercial product. Both direct spray assays and residual contact assays on treated cabbage leaf disks were carried out. The tested products had low efficacy against aphids when compared to pyrethrins but were in general less detrimental to ladybird beetle larvae, meaning that if applied against other pests, they have a lower chance of harming this agent of aphid biocontrol. Some of the products (soap, orange EO) did, however, show direct exposure toxicity toward ladybird larvae, and thyme EO had extensive phytotoxic effects on cabbage leaves, possibly indirectly leading to higher mortality in ladybird adults. These results underline the necessity for case-by-case evaluations of botanical insecticides.

Clinical Evidence on Expansion of Essential Oil-Based Formulation's Pharmacological Activity in Bovine Mastitis Treatment: Antifungal Potential as Added Value.

Bovine mastitis, as a significant and prevalent health problem in dairy herds, is primarily attributed to bacterial infections. Yeasts, although considered relatively rare causative agents, have also been associated with mastitis in dairy cattle. Current mastitis treatment predominantly relies on antibiotics, with limited emphasis on antifungal treatment. However, mycotic mastitis is challenging to treat, since these fungi are often resistant to antibiotics and may even utilize them for energy. In the current research, the in vivo antimicrobial activity of the essential oil-based formulation (Phyto-Bomat), as a possible alternative mastitis treatment associated with yeasts, was studied. This study involved a total of 68 animals from two dairy farms with diagnosed mastitis, and three treatment groups were established: conventional antibiotic treatment, Phyto-Bomat treatment, and the combination of both. The findings suggest significant variations in the presence of Candida samples based on the treatment administered, and the most significant difference was noted in cows treated with the combination (Phyto-Bomat and antibiotics). Yet, it is important to note that the results reveal that, regardless of the treatment type, there are statistically significant differences in the presence of Candida samples across the examined time points. These results aim to provide valuable insights into the potential of EOs as an alternative therapy in bovine mastitis, specifically targeting yeasts. Such findings could offer new strategies in the mycotic mastitis control and reducing the occurrence of secondary infections following antibiotic treatment.

Essential Oils from Southern Italian Aromatic Plants Synergize with Antibiotics against Escherichia coli, Pseudomonas aeruginosa and Enterococcus faecalis Cell Growth and Biofilm Formation.

The spread of antibiotic-resistant pathogens has prompted the development of novel approaches to identify molecules that synergize with antibiotics to enhance their efficacy. This study aimed to investigate the effects of ten Essential Oils (EOs) on the activity of nine antibiotics in influencing growth and biofilm formation in Escherichia coli, Pseudomonas aeruginosa, and Enterococcus faecalis. The effects of the EOs alone and in combination with antibiotics on both bacterial growth and biofilm formation were analyzed by measuring the MIC values through the broth microdilution method and the crystal violet assay, respectively. All EOs inhibited the growth of E. coli (1.25 ≤ MIC ≤ 5 mg/mL) while the growth of P. aeruginosa and E. faecalis was only affected by EOs from Origanum vulgare, (MIC = 5 mg/mL) and O. vulgare (MIC = 1.25 mg/mL) and Salvia rosmarinus (MIC = 5 mg/mL), respectively. In E. coli, most EOs induced a four- to sixteen-fold reduction in the MIC values of ampicillin, ciprofloxacin, ceftriaxone, gentamicin, and streptomycin, while in E. faecalis such a reduction is observed in combinations of ciprofloxacin with C. nepeta, C. bergamia, C. limon, C. reticulata, and F. vulgare, of gentamicin with O. vulgare, and of tetracycline with C. limon and O. vulgare. A smaller effect was observed in P. aeruginosa, in which only C. bergamia reduced the concentration of tetracycline four-fold. EO-antibiotic combinations also inhibit the biofilm formation. More precisely, all EOs with ciprofloxacin in E. coli, tetracycline in P. aeruginosa, and gentamicin in E. faecalis showed the highest percentage of inhibition. Combinations induce up- and down-methylation of cytosines and adenines compared to EO or antibiotics alone. The study provides evidence about the role of EOs in enhancing the action of antibiotics by influencing key processes involved in resistance mechanisms such as biofilm formation and epigenetic changes. Synergistic interactions should be effectively considered in dealing with pathogenic microorganisms.

Coatings Based on Essential Oils for Combating Antibiotic Resistance.

In the current era of widespread antimicrobial resistance, the utilization of essential oils (EOs) derived from plants has emerged as a promising alternative in combating pathogens that have developed resistance to antibiotics. This review explores the therapeutic potential of essential oils as valuable tools in restoring the efficacy of antibiotics, highlighting their unique ability to affect bacteria in multiple ways and target various cellular systems. Despite the challenge of elucidating their precise mode of action, EOs have shown remarkable results in rigorous testing against a diverse range of bacteria. This review explores the multifaceted role of EOs in combating bacterial microorganisms, emphasizing their extraction methods, mechanisms of action, and comparative efficacy against synthetic antibiotics. Key findings underscore the unique strategies EOs deploy to counter bacteria, highlighting significant differences from conventional antibiotics. The review extends to advanced coating solutions for medical devices, exploring the integration of EO formulations into these coatings. Challenges in developing effective EO coatings are addressed, along with various innovative approaches for their implementation. An evaluation of these EO coatings reveals their potential as formidable alternatives to traditional antibacterial agents in medical device applications. This renaissance in exploring natural remedies emphasizes the need to combine traditional wisdom with modern scientific advancements to address the urgent need for effective antimicrobial solutions in the post-antibiotic era.

Essential Oil of Fractionated Oregano as Motility Inhibitor of Bacteria Associated with Urinary Tract Infections.

In this research, several analyses were carried out on concentrated fractions of Mexican oregano essential oil (Poliomintha longiflora Gray) in order to determine its ability to inhibit the growth and the motility of Escherichia coli (swimming), Pseudomonas aeruginosa (swimming), and Proteus vulgaris (swarming); these Gram-negative bacteria associated with urinary tract infections are motile due to the presence of flagella, which is considered an important virulence factor that favors their motility when trying to reach the target organ and cause an infection. Also, the resistance pattern to antibiotics of each strain was determined. The results showed resistance pattern (8 out of 12 antibiotics tested) for P. aureginosa, while E. coli and P. vulgaris were resistant to 4 antibiotics out of the 12 tested. On the other hand, fractionated oregano caused an inhibition of growth and a reduction in motility, varying between fractions and among bacteria. Fraction 4 showed major growth reduction, with MBC values ranging from 0.002 to 23.7 mg/mL. Treatment with fractionated oregano (F1, F2, F3, F4) reduced the motility by 92-81% for P. vulgaris, 90-83% for E. coli, and 100-8.9% for P. aeruginosa. These results demonstrated a higher performance with a lower application dose due to its high content of Carvacrol and Thymol; unlike other concentrated fractions, this synergy of oxygenated monoterpenes may cause greater antimicrobial activity.

Synergistic Antimicrobial Activity of Biogenic Silver Nanoparticles and Acanthospermum australe Essential Oil against Skin Infection Pathogens.

In response to the steady increase in antimicrobial-resistant strains, the World Health Organisation has emphasised the need to investigate new antimicrobial agents and alternative therapies that improve the spectrum of activity and reduce the dose required, thus improving safety. This study focused on the characterisation of Acanthospermum australe essential oil and green-synthesis silver nanoparticles (AgNP), evaluating their cytotoxicity in human cells, antimicrobial activity and synergistic effect against pathogens causing skin infections. The main components of the essential oil were germacrene A (24.07%), γ-cadinene (21.47%) and trans-caryophyllene (14.97%). Spherical AgNP with a diameter of 15 ± 3 nm were synthesised. The essential oil showed antimicrobial activity against dermatophytes and Malassezia globosa, while AgNP were found to be active against bacteria, yeasts and dermatophytes. Both compounds were found to be primarily non-cytotoxic at the concentrations required to inhibit microbial growth. Furthermore, the combined use of essential oil and AgNP showed a synergistic antimicrobial effect against dermatophytes and M. globosa. In conclusion, the results suggest that the combined use of bioactive compounds from natural sources, such as essential oil and biogenic AgNP, has the potential to improve antimicrobial efficacy against specific skin pathogens, particularly Microsporum canis, Nannizzia gypsea and M. globosa.

Unveiling the mechanism of essential oil action against skin pathogens: from ancient wisdom to modern science.

Essential oils are among the most well-known phyto-compounds, and since ancient times, they have been utilized in medicine. Over 100 essential oils have been identified and utilized as therapies for various skin infections and related ailments. While numerous commercial medicines are available in different dosage forms to treat skin diseases, the persisting issues include their side effects, toxicity, and low efficacy. As a result, researchers are seeking novel classes of compounds as substitutes for synthetic drugs, aiming for minimal side effects, no toxicity, and high efficacy. Essential oils have shown promising antimicrobial activity against skin-associated pathogens. This review presents essential knowledge and scientific information regarding essential oil's antimicrobial capabilities against microorganisms that cause skin infections. Essential oils mechanisms against different pathogens have also been explored. Many essential oils exhibit promising activity against various microbes, which has been qualitatively assessed using the agar disc diffusion experiment, followed by determining the minimum inhibitory concentration for quantitative evaluation. It has been observed that Staphylococcus aureus and Candida albicans have been extensively researched in the context of skin-related infections and their antimicrobial activity, including established modes of action. In contrast, other skin pathogens such as Staphylococcus epidermidis, Streptococcus pyogens, Propionibacterium acnes, and Malassezia furfur have received less attention or neglected. This review report provides an updated understanding of the mechanisms of action of various essential oils with antimicrobial properties. This review explores the anti-infectious activity and mode of action of essential against distinct skin pathogens. Such knowledge can be valuable in treating skin infections and related ailments.

An Ethnopharmacological Survey on Medicinally Important Plants of Genus Salvia.

Lamiaceae (Labiatae) is a medicinally significant plant family featuring key species like Salvia aegyptiaca, S. cabulica, S. coccinea, S. glutinosa, S. officinalis, S. haematodes, S. hians, S. lanata, S. macrosiphon, S. moorcroftiana, S. spinosa, S. sclarea, and S. plebeia. These species exhibit diverse pharmacological activities attributed to essential oils and phytochemi-cals, including antioxidant, antiasthmatic, antitumor, anti-inflammatory, analgesic, etc. This re-view covers extensive phytomedicinal aspects of some important plants of the genus Salvia.

Comparative Anti-Cancer and Anti-Inflammatory Activities of Essential Oils from the Bark and Flower of Magnolia officinalis Rehd. et Wils.

This study was designed to compare the antioxidant, antitumor and anti-inflammatory effects of essential oils from the bark and flower of Magnolia officinalis Rehd. et Wils. Distillation extraction and steam distillation were used to extract EOs from the bark and flower. The results showed that the contents of EOs of SDE-F and SDE-B were much higher than that of SD-F and SD-B. EOs from the bark were rich in eudesmol (especially α-eudesmol) and exhibited a stronger antioxidant effect than the flower. The anti-tumor effects of SD-B and SD-F on HepG2 and MDA-MB-231 cells were better than that of SDE-B and SDE-F. The inhibitory rates of SD-B and SD-F on MDA-MB-231 cells were 59.21% and 48.27%, exceeding that of positive control 5-fluorouracil (47.04%) at 50 µg/mL. All four EOs exhibited excellent anti-inflammatory activities through the regulation of nitric oxide production and pro-inflammation cytokines in LPS-induced RAW 264.7 cells and they also remarkably suppressed the mRNA expressions of nitric oxide synthase, IL-6 and TNF-α at the concentration higher than that of positive control dexamethasone. These results indicated significant differences in the composition, and anti-inflammatory and anti-tumor activities of EOs extracted by different methods and provided a theoretical basis for their development and utilization.

Effect of Tea Tree Essential Oil on the Quality, Antioxidant Activity, and Microbiological Safety of Lightly Processed Lily (Lilium brownii var. viridulum) during Storage.

The Lanzhou lily is a regionally distinctive vegetable; the emergence of lightly processed lilies has addressed the inconvenience of consuming fresh lilies. However, the cleaning and impurity removal during the processing of lightly processed lily may strip off its original protective barrier and affect the edible quality. As one of the preservation methods, tea tree essential oil (TTEO) has the characteristics of being green, safe, and efficient preservative properties. This study focused on investigating the effects of different concentrations (25 µL/L, 50 µL/L, and 100 µL/L) of TTEO on the quality and microbiological safety of lightly processed lily. The results showed that compared with the control, appropriate concentrations of TTEO treatment could delay weight loss, improve appearance, firmness, and sensory quality, and maintain microbiological safety with the best effect observed at 50 µL/L. Meanwhile, TTEO treatment induced phenylalanine ammonia-lyase activity, thereby increasing the total phenolic content. Furthermore, TTEO enhanced the superoxide dismutase (SOD) and ascorbate peroxidase (APX) activity, which reduced O2-· production rate and H2O2 content. TTEO inhibited lipoxygenase (LOX) activity, reducing the relative conductivity and malondialdehyde content, thereby delaying lipid peroxidation and quality deterioration. This indicates that TTEO could enhance antioxidant capacity by regulating reactive oxygen species (ROS) metabolism and delay the quality deterioration of lightly processed lily by inhibiting lipid peroxidation.

Molecular and Phytochemical Characteristics of Flower Color and Scent Compounds in Dog Rose (Rosa canina L.).

This study delves into the chemical and genetic determinants of petal color and fragrance in Rosa canina L., a wild rose species prized for its pharmacological and cosmetic uses. Comparative analysis of white and dark pink R. canina flowers revealed that the former harbors significantly higher levels of total phenolics (TPC) and flavonoids (TFC), while the latter is distinguished by elevated total anthocyanins (TAC). Essential oils in the petals were predominantly composed of aliphatic hydrocarbons, with phenolic content chiefly constituted by flavonols and anthocyanins. Notably, gene expression analysis showed an upregulation in most genes associated with petal color and scent biosynthesis in white buds compared to dark pink open flowers. However, anthocyanin synthase (ANS) and its regulatory gene RhMYB1 exhibited comparable expression levels across both flower hues. LC-MS profiling identified Rutin, kaempferol, quercetin, and their derivatives as key flavonoid constituents, alongside cyanidin and delphinidin as the primary anthocyanin compounds. The findings suggest a potential feedback inhibition of anthocyanin biosynthesis in white flowers. These insights pave the way for the targeted enhancement of R. canina floral traits through metabolic and genetic engineering strategies.

Insecticidal Activity of Some Major Essential Oil Components against Metopolophium dirhodum and Its Predators.

Essential oils (EOs) are plant metabolites with important insecticidal effects. Nevertheless, information on the efficacy of the major substances on aphids and their natural enemies is still missing. The objective of this paper is, therefore, to identify the efficacy of selected EO majority substances-ß-citronellol, carvacrol, isoeugenol, and linalool, including their binary mixtures-on the mortality and fertility of the aphid Metopolophium dirhodum, an important cereal pest. The best efficacy was proven for the binary mixture of ß-citronellol and linalool (1:1 ratio), for which the estimated LC50(90) is 0.56(1.58) mL L-1. This binary mixture applied in sublethal concentrations significantly reduced aphid fertility. It was found that the phenomenon can be attributed to ß-citronellol, as the females treated with LC30 laid 45.9% fewer nymphs, on average, compared to the control. Although ß-citronellol and linalool, including their 1:1 mixture, showed very good efficacy on aphid mortality, they were, on the other hand, very friendly to the larvae of Aphidoletes aphidimyza and Chrysoperla carnea, which are important aphid predators. Based on our results, the newly discovered synergically acting binary mixture ß-citronellol/linalool can be recommended as an efficient substance suitable for the further development of botanical insecticides used against aphids.

THE COMBINATORY EFFECTS OF ESSENTIAL OIL FROM Lippia macrophylla ON MULTIDRUG RESISTANT Acinetobacter baumannii CLINICAL ISOLATES.

To assess the antibacterial effectiveness of Lippia macrophylla essential oil (LMEO) against multidrug-resistant Acinetobacter baumannii isolates, both as a standalone treatment and in combination with conventional antibiotics. LMEO demonstrated a significant inhibitory effect on the growth of A. baumannii, with a minimum inhibitory concentration (MIC) below 500µg/mL. Notably, LMEO was capable of reversing the antibiotic resistance of clinical isolates or reducing their MIC values when used in combination with antibiotics, showing synergistic (FICI ≤ 0.5) or additive effects. The combination of LMEO and imipenem was particularly effective, displaying synergistic interactions for most isolates. Ultrastructural analyses supported these findings, revealing that the combination of LMEO + ceftazidime compromised the membrane integrity of the Acb35 isolate, leading to cytoplasmic leakage and increased formation of Outer Membrane Vesicles (OMVs). Taken together our results point for the use of LMEO alone or in combination as an antibacterial agent against A. baumannii. These findings offer promising avenues for utilizing LMEO as a novel antibacterial strategy against drug-resistant infections in healthcare settings, underscoring the potential of essential oils in enhancing antibiotic efficacy.

Optimization of Enzyme-Assisted Extraction of Rosemary Essential Oil Using Response Surface Methodology and Its Antioxidant Activity by Activating Nrf2 Signaling Pathway.

Rosemary essential oil (REO) is widely recognized as a food flavoring and traditional herb and possesses potential antioxidant activity. However, its low yield rate and unclarified antioxidant mechanism warrant further investigation. In this study, an enzyme pretreatment-assisted extraction method with Box-Behnken design (BBD) and response surface methodology (RSM) models was employed to optimize the main factors of REO, and its antioxidant molecular mechanism under oxidative stress was elucidated in hydrogen peroxide-induced human lung carcinoma (A549) cells. The optimized yield (4.10%) of REO was recorded with the following optimum conditions: enzyme amount 1.60%, enzyme digestion pH 5.0, enzyme digestion temperature 46.50 °C, and enzyme digestion time 1.7 h. Meanwhile, 1.8-cineole (53.48%) and ß-pinene (20.23%) exhibited radical scavenging activity higher than that of BHA and BHT. At the cellular level, REO (12.5-50 µg/mL) increased the levels of cell viability, CAT, SOD, and GSH significantly while reducing the contents of ROS, MDA, and GSSG, when compared to H2O2 exposure. Mechanically, REO relieved oxidative stress via activating the Nrf2 signaling pathway and enhancing the protein expression of Nrf2, NQO-1, and HO-1, which was further verified by molecular docking between the main component 1.8-cineole and the Kelch domain of KEAP1. Therefore, REO could be considered as a potent natural antioxidant with a potential strategy in the food and pharmaceutical industries.

Imiquimod-Loaded Nanosystem for Treatment Human Papillomavirus-Induced Lesions.

Human papillomavirus (HPV)-associated cervical cancer is the most common cancer among women worldwide. The treatment options are strongly related to increased infertility in women. Imiquimod (IQ) is an imidazoquinoline, which has proven antiviral effects against persistent HPV infection by activating immune cells via Toll-like receptors 7/8 when formulated in carriers, like nanogels, for topical use. An effective alternative to conventional therapies is the nanoparticle drug delivery system. We studied lipidic nanoparticles with IQ (Lipo IQ) and functionalized them with a DNA aptamer, AT11 (Lipo IQ AT11), to improve the selectivity for cervical cancer cells combined with the efficacy of essential oils. The formulations showed that the physicochemical properties are adequate for vaginal drug delivery and have antimicrobial activity at higher concentrations (with MIC50 starting from 0.625%). The final formulations exhibited cytotoxicity in cancer cells, enhanced by essential oils without affecting healthy cells, resulting in less than 10% cell viability in HeLa cells and over 60% in NHDF cells. Essential oils potentiate Lipo IQ's effectiveness, while AT11 increases the selectivity for cervical cancer cells. As suggested by the results of the permeation assay, the formulations were internalized by the cancer cells. Overall, the obtained results suggested that the synergistic effect of the essential oils and the nanosystem potentiate the cytotoxic effect of Lipo IQ and that Lipo IQ AT11 promotes selectivity towards cancer cells.

Synergistic Effect of Essential Oils and Antifungal Agents in Fighting Resistant Clinical Isolates of Candida auris.

Recently, a large number of nosocomial infections have been caused by an emerging pathogen that is rising as a worldwide issue in human health: Candida auris. This yeast is considered resistant to antifungals of the first-line therapies, and consequently it is related to morbidity and mortality. Therefore, the aim of this research was to determine the in vitro anti-C. auris activity against twenty-three resistant clinical strains of different essential oils (EOs), pure or in combination with traditional antifungal agents, mainly caspofungin, fluconazole, micafungin and 5-flucytosine. Broth dilution assay was performed to evaluate the fungistatic and fungicidal effectiveness of fifteen EOs towards all the C. auris isolates. The data demonstrated that EOs were able to prevent C. auris growth, with MIC values ranging from 0.03 to 1% for the efficacious EOs (thyme, cinnamon, geranium, clove bud, lemongrass and mentha of Pancalieri), whereas the MICs were >1% for the ineffective ones. Thereafter, the six most effective EOs were used to perform the checkerboard experiments by assaying simultaneously the activity of EOs and traditional antifungals towards two selected strains. The most promising synergic combinations towards C. auris, depending on the isolate, were those with micafungin and geranium, thyme, cinnamon, lemongrass or clove bud EOs, with fluconazole and mentha of Pancalieri EO, and with 5-flucytosine and mentha of Pancalieri EO. These EOs and their combinations with antifungal drugs may provide a useful therapeutic alternative that could reduce the dose of the individual components, limiting the overall side effects. These associations might be a prospective option for the future treatment of infections, thus helping to overcome the challenging issue of resistance in C. auris.

Salvia verticillata (L.)-Biological Activity, Chemical Profile, and Future Perspectives.

Species belonging to the genus Salvia, Lamiaceae, have been deeply involved in the folk medicine of different nations since ancient times. Lilac sage, or Salvia verticillata L. (S. verticillata) is a less studied species from the genus. However, it seems to have a prominent potential for the future drug discovery strategies of novel phytopharmaceuticals. This review aims to summarise the data on the biological activity and the phytochemical profile of extracts and essential oils derived from S. verticillata. This review is based on data from 57 in vitro and in vivo studies. The chemical profile of S. verticillata includes different synergic compounds like phenolic acids, flavonoids, terpenes, and salvianolic acids. Although some small amounts of salvianolic acid B were found in S. verticillata extracts, the major compound among the salvianolic acids is salvianolic acid C, a compound associated with the potential for improving liver fibrosis, cardio- and hepatoprotection, and the inhibition of SARS-CoV-2 infection. The cannabinoid type 2 receptor agonist ß-caryophyllene is one of the major compounds in S. verticillata essential oils. It is a compound with a prominent potential in regenerative medicine, neurology, immunology, and other medical fields. The in vivo and the in vitro studies, regarding S. verticillata highlighted good antioxidant potential, anti-inflammatory, antibacterial, and antifungal activity. S.verticillata was also reported as a potential source of drug candidates for the treatment of neurodegenerative diseases such as Alzheimer's disease, because of the inhibitory activity on the acetylcholinesterase. However, the number of studies in this direction is limited.

Phytochemical Analysis and Antioxidant and Antifungal Activities of Powders, Methanol Extracts, and Essential Oils from Rosmarinus officinalis L. and Thymus ciliatus Desf. Benth.

Chemical residues in food pose health risks such as cancer and liver issues. This has driven the search for safer natural alternatives to synthetic fungicides and preservatives. The aim of this study was to characterize the chemical composition of the essential oils (EO), determine the polyphenolic contents, and evaluate the in vitro antioxidant and antifungal activities of methanol extracts (ME), essential oils (EO), and powders from Rosmarinus officinalis L. (rosemary) and Thymus ciliatus (Desf) Benth. (thyme) from the M'sila region, Algeria. The chemical composition of the EOs was determined by GC-MS. R. officinalis EO was composed of 31 components, mainly camphor (41.22%), camphene (18.14%), and α-pinene (17.49%); T. ciliatus EO was composed of 58 components, mainly, in percentage, α-pinene (22.18), myrcene (13.13), ß-pinene (7.73), ß-caryophyllene (10.21), and germacrene D (9.90). The total phenols and flavonoids were determined spectrophotometrically, and the rosemary ME was found to possess the highest polyphenolic content (127.1 ± 2.40 µg GAE/mg), while the thyme ME had the highest flavonoid content (48.01 ± 0.99 µg QE/mg). The antioxidant activity was assessed using three methods: rosemary ME was the most potent, followed by DPPH (IC50 = 13.43 ± 0.14 µg/mL), ß-carotene/linoleic acid (IC50 = 39.01 ± 2.16 µg/mL), and reducing power (EC50 = 15.03 ± 1.43 µg/mL). Antifungal activity was assessed for 32 pathogenic and foodborne fungi. Four methods were applied to the solid medium. Incorporating the powdered plant into the culture medium (at 10%) reduced the fungal growth to greater than 50% in 21.88% and 6.25% of all fungal isolates, for R. officinalis and T. ciliatus, respectively. The ME, applied by the well diffusion method (0.1 g/mL), was less effective. Different concentrations of EO were tested. Incorporating the EO into the culture medium (1500 µL/L) inhibited 50% of the molds to levels of 50 and 75% for R. officinalis and T. ciliatus, respectively, with the complete inhibition of four fungi. Fumigated EO (15 µL) inhibited 65% of the molds to levels of 65 and 81.25% for R. officinalis and T. ciliatus, respectively, with the complete inhibition of five fungi. There was little to no sporulation in conjunction with the inhibition. Our results revealed some of the potential of the studied plants to fight foodborne molds and presented their promising characteristics as a source of alternatives to chemical pesticides and synthetic preservatives. Further studies are needed to find adequate application techniques in the food safety area.