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Background: In recent decades cyanobacterial species have attracted research attention as potential sources of new biostimulants. In this study, the biostimulant effects of five cyanobacterial suspensions on the growth and essential oil composition of Thymus vulgaris L. were evaluated. The expression of key genes involved in the biosynthesis of thymol and carvacrol, such as DXR and TPS2, were investigated., Results: A pot culture experiment revealed that cyanobacterial application significantly improved T. vulgaris L. growth indices, including plant height, dry and fresh weight, leaf and flower number, leaf area, and photosynthetic pigment content. Total phenol and flavonoid content in inoculated plants also showed a significant increase compared with the control. Anabaena torulosa ISB213 inoculation significantly increased root and shoot biomass by about 65.38% and 92.98% compared with the control, respectively. Nostoc calcicola ISB215 inoculation resulted in the highest amount of essential oil accumulation (18.08 ± 0.62) in T. vulgaris leaves, by about 72.19% compared with the control (10.5 ± 0.50%). Interestingly, the amount of limonene in the Nostoc ellipsosporum ISB217 treatment (1.67%) increased significantly compared with the control and other treatments. The highest expression rates of DXR and TPS2 genes were observed in the treatment of N. ellipsosporum ISB217, with 5.92-fold and 5.22-fold increases over the control, respectively., Conclusion: This research revealed the potential of the cyanobacteria that were studied as promising biostimulants to increase the production of biomass and secondary metabolites of T. vulgaris L., which could be a suitable alternative to chemical fertilizers. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

The research investigates the influence of different lighting conditions and soil treatments, in particular the application of food polymers separately and in combination with spores of Trichoderma consortium, on the growth and development of herbs- Thymus vulgaris and Thymus serpyllum . The metabolic analysis focuses on detecting changes in the levels of biologically active compounds such as chlorophyll a and b, anthocyanins, carotenoids, phenolic compounds (including flavonoids), terpenoids, and volatile organic compounds with potential health-promoting properties. By investigating these factors, the study aims to provide insights into how environmental conditions affect the growth and chemical composition of selected plants and to shed light on potential strategies for optimising the cultivation of these herbs for the improved quality and production of bioactive compounds. Under the influence of additional lighting, the growth of T. vulgaris and T. serpyllum seedlings was greatly accelerated, resulting in an increase in shoot biomass and length, and in the case of T. vulgaris , an increase in carotenoid and anthocyanin contents. Regarding secondary metabolites, the most pronounced changes were observed in total antioxidant capacity and flavonoid content, which increased significantly under the influence of additional lighting. The simultaneous or separate application of Trichoderma and food polymers resulted in an increase in flavonoid content in the leaves of both Thymus species. The increase in terpenoid content under supplemental light appears to be related to the presence of Trichoderma spores as well as food polymers added to the soil. However, the nature of these changes depends on the thyme species. Volatile compounds were analysed using an electronic nose (E-nose). Eight volatile compounds (VOCs) were tentatively identified in the vapours of T. vulgaris and T. serpyllum : α-pinene, myrcene, α-terpinene, γ-terpinene; 1,8-cineole (eucalyptol), thymol, carvacrol, and eugenol. Tendencies to increase the percentage of thymol and γ-terpinene under supplemental lighting were observed. The results also demonstrate a positive effect of food polymers and, to a lesser extent, Trichoderma fungi on the synthesis of VOCs with health-promoting properties. The effect of Trichoderma and food polymers on individual VOCs was positive in some cases for thymol and γ-terpinene., Competing Interests: The authors declare no conflicts of interest.

Arsenic (As) is a highly cytotoxic element impairing normal cellular functions, and its bioremediation has become one of the environmental concerns. This study explored the molecular and physiological responses of thyme (Thymus kotschyanus) seedlings to incorporating As (0 and 10 mgl -1 ) and methyl jasmonate (MJ; 0 and 10 µM) into the culture medium. The MJ treatment reinforced root system and mitigated the As cytotoxicity risk. MJ contributed to hypomethylation, a potential adaptation mechanism for conferring the As tolerance. Two cytochrome P450 monooxygenases, including CYP71D178 and CYP71D180 genes, were upregulated in response to As and MJ. The MJ treatment contributed to up-regulation in the γ-terpinene synthase (TPS) gene, a marker gene in the terpenoid metabolism. The As presence reduced photosynthetic pigments (chlorophylls and carotenoids), while the MJ utilization alleviated the As toxicity. The MJ supplementation increased proline accumulation and soluble phenols. The application of MJ declined the toxicity sign of As on the concentration of proteins. The activities of peroxidase, catalase, and phenylalanine ammonia-lyase (PAL) enzymes displayed an upward trend in response to As and MJ treatments. Taken collective, MJ can confer the As tolerance by triggering DNA hypomethylation, regulating CYPs, and stimulating primary and secondary metabolism, especially terpenoid., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Water and ethanol extracts of dried thyme ( Thymus vulgaris ) were analyzed for chemical composition, inhibition of the SARS-CoV-2 spike protein-ACE2 interaction, inhibition of ACE2 activity, and free radical scavenging capacity. Thirty-two compounds were identified in water extract (WE) and 27 were identified in ethanol extract (EE) of thyme through HPLC-MS. The WE (33.3 mg/mL) and EE (3.3 mg/mL) of thyme inhibited the spike protein-ACE2 interaction by 82.6 and 86.4%, respectively. The thyme WE at 5 mg/mL inhibited ACE2 activity by 99%, and the EE at 5 mg/mL inhibited ACE2 by 65.8%. Total phenolics were determined to be 38.9 and 8.8 mg of GAE/g in WE and EE, respectively. The HO • scavenging capacities were 1121.1 and 284.4 μmol of TE/g in WE and EE, respectively. The relative DPPH • scavenging capacities were 126.3 μmol TE/g in WE and 28.2 μmol TE/g in EE. The ABTS •+ scavenging capacities were 267.1 μmol TE/g in WE and 96.7 μmol TE/g in EE. The results suggested that the thyme extract could be potentially used to prevent SARS-CoV-2 infection and mitigate the complications from the infection.

Background: Thyme derived essential oil and its components have numerous applications in pharmaceutical, food, and cosmetic industries, owing to their antibacterial, antifungal, and antiviral properties. To obtain thyme essential oil with different terpene composition, we developed new germplasm resources using the conventional hybridization approach., Results: Phenotypic characteristics, including essential oil yield and composition, glandular trichome density, plant type, and fertility, of three wild Chinese and seven European thyme species were evaluated. Male-sterile and male-fertile thyme species were crossed in different combinations, and two F 1 populations derived from Thymus longicaulis (Tl) × T. vulgaris 'Fragrantissimus' (Tvf) and T. vulgaris 'Elsbeth' (Tve) × T. quinquecostatus (Tq) crosses were selected, with essential oil yield and terpene content as the main breeding goals. Simultaneously, simple sequence repeat (SSR) primers were developed based on the whole-genome sequence of T. quinquecostatus to authenticate the F 1 hybrids. A total of 300 primer pairs were selected, and polymerase chain reaction (PCR) was carried out on the parents of the two hybrid populations (Tl, Tvf, Tve, and Tq). Based on the chemotype of the parents and their F 1 progenies, we examined the expression of genes encoding two γ-terpinene synthases, one α-terpineol synthase, and maybe one geraniol synthase in all genotypes by quantitative real-time PCR (qRT-PCR)., Conclusion: We used hybridization to create new germplasm resources of thyme, developed SSR markers based on the whole-genome sequence of T. quinquecostatus, and screened the expression of monoterpene synthase genes in thyme. The results of this study provide a strong foundation for the creation of new germplasm resources, construction of the genetic linkage maps, and identification of quantitative trait loci (QTLs), and help gain insight into the mechanism of monoterpenoids biosynthesis in thyme., (© 2023. The Author(s).)

Thyme has medicinal and aromatic value because of its potent antimicrobial and antioxidant properties. However, the absence of a fully sequenced thyme genome limits functional genomic studies of Chinese native thymes. Thymus quinquecostatus Čelak., which contains large amounts of bioactive monoterpenes such as thymol and carvacrol, is an important wild medicinal and aromatic plant in China. Monoterpenoids are abundant in glandular secretory trichomes. Here, high-fidelity and chromatin conformation capture technologies were used to assemble and annotate the T. quinquecostatus genome at the chromosome level. The 13 chromosomes of T. quinquecostatus had a total length of 528.66 Mb, a contig N50 of 8.06 Mb, and a BUSCO score of 97.34%. We found that T. quinquecostatus had experienced two whole-genome duplications, with the most recent event occurring ∼4.34 million years ago. Deep analyses of the genome, in conjunction with comparative genomic, phylogenetic, transcriptomic, and metabonomic studies, uncovered many regulatory factors and genes related to monoterpenoids and glandular secretory trichome development. Genes encoding terpene synthase (TPS), cytochrome P450 monooxygenases (CYPs), short-chain dehydrogenase/reductase (SDR), R2R3-MYB, and homeodomain-leucine zipper (HD-ZIP) IV were among those present in the T. quinquecostatus genome. Notably, Tq02G002290.1 (TqTPS1) was shown to encode the terpene synthase responsible for catalyzing production of the main monoterpene product γ-terpinene from geranyl diphosphate (GPP). Our study provides significant insight into the mechanisms of glandular secretory trichome formation and monoterpenoid biosynthesis in thyme. This work will facilitate the development of molecular breeding tools to enhance the production of bioactive secondary metabolites in Lamiaceae., (Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.)

Background: Titanium dioxide nanoparticles (TiO 2 -NPs) are widely utilized in medicine and industry; however, their safety in biological organisms is still unclear. In this study, we determined the bioactive constitutes of thyme essential oil (TEO) and utilized the nanoemulsion technique to improve its protective efficiency against oxidative stress, genotoxicity, and DNA damage of biosynthesized titanium dioxide nanoparticles (TiO 2 -NPs)., Methods: TEO nanoemulsion (TEON) was prepared using whey protein isolate (WPI). Sixty male Sprague-Dawley rats were divided into six groups and treated orally for 21 days including the control group, TEO, or TEON- treated groups (5 mg/kg b.w), TiO 2 -NPs-treated group (50 mg/kg b.w) and the groups received TiO 2 -NPs plus TEO or TEON. Blood and tissues samples were collected for different assays., Results: The GC-MS analysis identified 17 bioactive compounds in TEO and thymol and carvacrol were the major compounds. TEON was irregular with average particles size of 230 ± 3.7 nm and ζ-potential of -24.17 mV. However, TiO2-NPs showed a polygonal shape with an average size of 50 ± 2.4 nm and ζ-potential of -30.44 mV. Animals that received TiO2-NPs showed severe disturbances in liver and kidney indices, lipid profile, oxidant/antioxidant indices, inflammatory cytokines, gene expressions, increased DNA damage, and pathological changes in hepatic tissue. Both TEO and TEON showed potential protection against these hazards and TEON was more effective than TEO., Conclusion: The nanoemulsion of TEO enhances the oil bioactivity, improves its antioxidant characteristics, and protects against oxidative damage and genotoxicity of TiO2-NPs., (Copyright © 2022 Elsevier GmbH. All rights reserved.)

Type 2 diabetes mellitus (T2DM) is a complex metabolic disorder that is characterized by hyperglycemia, insulin resistance, and lack of insulin production. It has been previously reported that Thymus serpyllum has therapeutic potential against many diseases. To investigate the antidiabetic action of Thymus serpyllum , this study aimed to analyze its restorative impact in diabetic mice, in which it was administered in diet. Diabetes was induced in BALB/c mice fed with a high-fat diet and two intraperitoneal injections of streptozotocin. With the onset of diabetes, the mice were administered daily with aqueous extract of Thymus serpyllum (500 mg/kg/d and 800 mg/kg/d) for 4 weeks. Body weight and fasting blood glucose levels were measured after every 1 week of the treatment. Subsequently, intraperitoneal glucose tolerance and insulin tolerance tests were conducted. In addition, liver tissue was isolated for assessment in terms of levels of gene expression of the AMPK , IRS1 , and GLUT2 gene. Treatment with the aqueous extract of Thymus serpyllum was found to be significantly effective in controlling hyperglycemia and improving glucose and insulin tolerance. Predictable with these impacts, the extract of Thymus serpyllum upregulated the AMPK expression at the mRNA level, as well as upregulating the expression of IRS1 and GLUT2 gene. Histopathological examination of the liver, kidney, and pancreas also revealed the restorative impact in terms of cellular morphology. The results hence demonstrated that oral administration of aqueous extract of Thymus serpyllum can potentially attenuate hyperglycemia in the liver muscle of streptozotocin (STZ)-induced diabetic mice via AMPK and IRS1 upregulation.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder with neuronal loss in the hippocampus. Our aim was to evaluate the effects of Iranian thyme honey (single dose: 2 gr/kg) vs rivastigmine (0.3 mg/kg) in vivo on spatial memory and in vitro on important parameters of oxidative stress as well as quantitative and qualitative studies of hippocampal neurons of AD rat models with this design that 30 days after oral administration of 17 mg/kg AlCl 3 , 20 AD rats were received that underwent a 6-weeks therapeutic period and another 20 rats underwent a 6-weeks preventive period and also 20 rats were as controls. Y-Maze test was performed to show memory deficiency as well as TBARS and FRAP assays to measure malondialdehyde (MDA) and total antioxidant, respectively. In addition, H&E staining was also done for cell counting and morphological changes. We observed that AD rats with hippocampal damage had more significant errors during the Y-maze test than the control and other rats. Likewise, MDA and neurodegeneration increased in the AD group while in all preventive and therapeutic group's especially Iranian thyme honey, they decreased and conversely, total antioxidant and number of normal cells elevated and healthy neurons were observed in all parts of the hippocampus and cortex. Our results despite the limitations showed the powerful antioxidant properties and cytoprotective effects of Iranian thyme honey vs rivastigmine on hippocampal neurons that consequently enhanced memory and if advanced diagnostic tests in human clinical patients show other more pronounced effects, we have certainly started a key and targeted strategy., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Humans often respond to sensory impulses provided by aromas, and current trends have generated interest in natural sources of fragrances rather than the commonly used synthetic additives. For the first time, the resulting aroma of a selected culture of Thymus mastichina L. was studied as a potential food ingredient. In this context, dried (DR) and fresh (FR) samples were submitted to carbon dioxide (CO 2 ) supercritical extraction (SFE) and hydrodistillation (HD) methods. The extracts were characterised according to their volatile composition by GC-MS, cytotoxicity against a non-tumour cell culture, and sensory attributes (odour threshold and olfactive descriptors). The most abundant aromas were quantified, and the analysis performed by GC-MS revealed an abundance of terpenoids such as thymol chemotype, followed by the precursors α-terpinene and p -cymene. DR and FR extracts (EX) obtained from SFE-CO 2 show the highest content of thymol, achieving 52.7% and 72.5% of the isolated volatile fraction. The DR essential oil (EO) contained the highest amount of terpenoids, but it was also the most cytotoxic extract. In contrast, SFE-CO 2 products showed the lowest cytotoxic potential. Regarding FR-OE, it had the lowest extraction yield and composition in aroma volatiles. Additionally, all samples were described as having green, fresh and floral sensory notes, with no significant statistical differences regarding the odour detection threshold (ODT) values. Finally, FR-EX of T. mastichina obtained by SFE-CO 2 presented the most promising results regarding food application.

The effects of plant-based extracts on the solar aging and antimicrobial properties of impregnated ethylene-norbornene (EN) copolymer and poly(lactic acid) (PLA) were investigated. In this study, the impregnation yield of polyolefin, lacking in active centers capable of phytochemical bonding, and polyester, abundant in active sides, was measured. Moreover, two different extracts plentiful in phytochemicals-thyme (TE) and clove (CE)-were employed in the solvent-based impregnation process. The effect of thymol and eugenol, the two main compounds embodied in the extracts, was studied as well. Interestingly, oxidation induction times (OIT) for the impregnation of EN with thyme and clove extracts were established to be, respectively, 27.7 and 39.02 min, which are higher than for thymol (18.4 min) and eugenol (21.1 min). Therefore, an aging experiment, mimicking the full spectrum of sunlight, was carried out to investigate the resistance to common radiation of materials impregnated with antioxidative substances. As expected, the experiment revealed that the natural extracts increased the shelf-life of the polymer matrix by inhibiting the degradation processes. The aging resistance was assessed based on detected changes in the materials' behavior and structure that were examined with Fourier-transform infrared spectroscopy, contact angle measurements, color quantification, tensile tests, and hardness investigation. Such broad results of solar aging regarding materials impregnated with thyme and clove extracts have not been reported to date. Moreover, CE was found to be the most effective modifying agent for enabling material with antimicrobial activity against Escherichia coli to be obtained.

The study of bioactive molecules of natural origin is a focus of current research. Thymus algeriensis and Artemisia herba-alba are two medicinal plants widely used by the Moroccan population in the traditional treatment of several pathologies linked to inflammation. This study aimed to evaluate the single and combined antioxidant, anti-inflammatory and analgesic effects of the essential oils extracted from these two medicinal plants, and also their potential toxicity. Essential oils were extracted using hydro-distillation in a Clevenger-type apparatus. The antioxidant activity was evaluated by two methods: the scavenging of the free radical DPPH, and the reduction in iron. Anti-inflammatory activity was evaluated by evaluating the edema development induced by carrageenan injecting, while the analgesic power was evaluated according to the number of abdominal contortions induced by the intraperitoneal injection of acetic acid (0.7%). The acute oral toxicity was performed to assess the potential toxicity of the studied EOs, followed by an analysis of the blood biochemical parameters. The results of the two antioxidant tests indicated that our extract mixture exhibits good iron reduction capacity and very interesting DPPH free radical scavenging power, with an IC 50 of around 4.38 ± 0.98 μg/mL higher than that of the benchmark antioxidant, BHT. The anti-inflammatory test demonstrated that the mixture administered orally at a dose of 150 mg/kg has a better activity, exceeding that of 1% Diclofenac, with a percentage of maximum inhibition of the edema of 89.99 ± 4.08. The number of cramps in the mice treated with the mixture at a dose of 150 mg/kg is significantly lower (29.80 ± 1.92) than those of the group treated with Tramadol (42.00 ± 2.70), respectively. The toxicity results show no signs of toxicity with an LD 50 greater than 150 mg/Kg. These interesting results show that the two plants' EOs had an important anti-inflammatory, analgesic, and antioxidant activity, and also a powerful synergistic effect, which encourages further in-depth investigations on their pharmacological proprieties.

Thyme species are a good source of thymol and carvacrol, which play a key role in controlling diseases. For the first time, the expression patterns of γ-terpinene synthase (TPS2), CYP71D178 , and CYP71D180 genes and the amount of phenolics compounds were evaluated in T. migricus and T. daenensis after different methyl jasmonate (MeJA) treatments. The highest thymol and carvacrol contents were observed in T. migricus (86.27%) and T. daenensis (17.87%) at MeJA 100 µM, which was consistent with the expression patterns of the three investigated genes. All species treated showed high total phenolic and flavonoid content compared to control plants for which the highest amounts were observed in T. vulgaris treated with 100 µM and 10 µM MeJA. Furthermore, in the 100 µM MeJA treatment, the relative expression of TPS2 and CYP71D178 in T. migricus increased 7.47 and 9.86-fold compared with the control, respectively. The highest level of CYP71D180 transcripts (5.15-fold) was also observed for T. daenensis treated. This finding highlights the notion that thymol was known as the dominant component of the essential oil rather than carvacrol in diffident thyme species. This implies that MeJA at different concentrations influenced metabolic pathways and induced expression changes, resulting in a rise in essential oil levels.

Intercropping of medicinal plants/legumes along with bio-fertilizer application is a relatively new sustainable practice for improving the yield and secondary metabolites production. Here, a 2-years field experiment was performed to evaluate the effects of water deficit stress and arbuscular mycorrhizal fungi (AMF) application (as bio-fertilizer) on nutrients concentration, dry matter yield, essential oil quantity and quality of thyme in intercropping with soybean. Three irrigation levels, including (i) irrigation after depletion of 20% (I 20 ) as non-stressed, 50% (I 50 ) as moderate water deficit and 80% (I 80 ) available water as severe water deficit were applied as the main factor. The sub-factor was represented by different cropping patterns including thyme sole culture, replacement intercrop ratio of 50:50 and 66:34 (soybean: thyme) and the third factor was non-usage (control) and usage of AMF. According to our results, the thyme dry yield under moderate and severe water deficit stress decreased by 35 and 44% in the first year, and by 27 and 40% in the second year compared with non-stressed (I 20 ) plants, respectively. Also, the macro- and micro-nutrients of thyme leaves increased significantly in intercropping patterns after application of AMF. The maximum essential oil percentage of thyme was achieved in 50:50 intercropping ratio treated with AMF. Under moderate and severe water deficits, the major constituents of thyme essential oil including thymol, p-cymene and γ-terpinene were increased in intercropping patterns treated with AMF. Generally, AMF application in intercropping ratio of 50:50 may be proposed to farmers as an eco-friendly approach to achieve desirable essential oil quality and quantity in thyme under water deficit stress conditions., (© 2021. The Author(s).)

This study aimed at an experimental design of response surface methodology (RSM) in the optimization of the dominant volatile fraction of Greek thyme honey using solid-phase microextraction (SPME) and analyzed by gas chromatography-mass spectrometry (GC-MS). For this purpose, a multiple response optimization was employed using desirability functions, which demand a search for optimal conditions for a set of responses simultaneously. A test set of eighty thyme honey samples were analyzed under the optimum conditions for validation of the proposed model. The optimized combination of isolation conditions was the temperature (60 °C), equilibration time (15 min), extraction time (30 min), magnetic stirrer speed (700 rpm), sample volume (6 mL), water: honey ratio (1:3 v / w ) with total desirability over 0.50. It was found that the magnetic stirrer speed, which has not been evaluated before, had a positive effect, especially in combination with other factors. The above-developed methodology proved to be effective in the optimization of isolation of specific volatile compounds from a difficult matrix, like honey. This study could be a good basis for the development of novel RSM for other monofloral honey samples.

In the present research, inter and intra genetic variability of 77 accessions belonging to 11 Thymus species were assessed using eight SRAP primer combinations. High polymorphism (98.3%) was observed in the studied species. The cluster analysis classified Thymus species into five main groups. According to molecular variance (AMOVA) analysis, 63.14% of total genetic variation was obtained within the species, while 36.86% of variation was observed among species. STRUCTURE analysis was also performed to estimate the admixture of species. For instance, T. carmanicus and T. transcaspicus revealed high admixtures. HPLC analysis also demonstrated the presence of rosmarinic acid (32.3-150.7 mg/100 g DW), salvianolic acid (8-90 mg/100 g DW), and cinnamic acid (1.7-32.3 mg/100 g DW) as major phenolic acids, as well as apigenin, epicatechin, and naringenin as the major flavonoids. The highest phenolic and flavonoid contents were detected in T. transcaspicus (37.62 mg gallic acid equivalents (GAE) g -1 DW) and T. vulgaris (8.72 mg quercetin equivalents (QE) g -1 DW), respectively. The antioxidant properties and total phenolic of Thymus species were examined using DPPH and β-carotene-linoleic acid model systems and consequently T. vulgaris and T. pubescens were detected with the highest and the lowest antioxidant activities respectively. Cluster and principal Components Analysis (PCA) of the components classified the species in to three groups. Finally, similarity within some species was observed comparing molecular and phytochemical markers. For instance, T. vulgaris separated from other species according to major polyphenolic profiles and molecular analyses, as well as T. transcaspicus, T. carmanicus, and T. fedtschenkoi that were clustered in the same groups.

This work has been aimed at studying the effect of red thyme oil (RTO, Thymus vulgaris L.) on the shelf-life and Penicillium decay of oranges during cold storage. RTO vapours significantly reduced (P ≤ 0.05) the percentage of infected wounds, the external growth area and the production of spores in inoculated orange fruit stored for 12 days at 7 °C in a polypropylene film selected for its appropriate permeability. Among the RTO compounds, p-cymene and thymol were the most abundant in packed boxes at the end of cold storage. The RTO vapours did not affect the main quality parameters of the oranges, or the taste and odour of the juice. The results have shown that an active packaging, using RTO vapours, could be employed, by the citrus industry, to extend the shelf-life of oranges for fresh market use and juice processing., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

In this study, the chemical composition, antimicrobial, antioxidant, and anticancer effects of Thymus convolutus Klokov oil and its main compound camphor were investigated. The oil was isolated from T. convolutus using hydrodistillation method, analyzed by gas chromatography/mass spectrometry (GC-MS), and 66 compounds were identified. The main component was determined as camphor at 16.6%. The antioxidant properties were identified with the DPPH (2,2'-diphenyl-1-picrylhydrazyl) radical-scavenging method and, 33.39 ± 0.25% DPPH was scavenging in 1000 μg/mL of essential oil. The strong antimicrobial activity was observed against Escherichia coli, Enterobacter aerogenes, Proteus vulgaris, and Pseudomonas aeruginosa with MIC values of 125 μg/mL. Aspergillus flavus was more sensitive (28%) against T. convolutus essential oil than other fungi. The cytotoxic effect of oil was analyzed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) method. Camphor was effective on human hepatoma cells (Hep3B) at concentrations of 1 mg/mL, 500, 250, and 125 μg/mL, while essential oil of T. convolutus was found to be effective at concentrations of 250 and 125 μg/mL. A reduction in cell proliferation was observed in colon carcinoma cells (HT-29) treated with 500 μg/mL camphor for 48 h. No statistically significant effect was found in Umbilical Vein Endothelial Cells (HUVEC) treated with essential oil and camphor., (© 2021 Walter de Gruyter GmbH, Berlin/Boston.)

We have previously demonstrated that the Thymus algeriensis and Thymus fontanesii extracts have powerful anti-inflammatory, antipyretic, and analgesic effects against acute pain models. We profiled their chemical composition and found many phenolic acids, flavonoids, and phenolic diterpenes. In this work, we investigated their antioxidant properties on HaCaT cells exposed to UVA-induced oxidative stress and examined their effects against chronic neuropathic pain and the underlying mechanisms. Through a rat chronic constriction injury (CCI) model, we induced chronic neuropathic pain by placing 4 loose ligatures around the right sciatic nerve for 14 days. Thermal and mechanical hyperalgesia in addition to cold and dynamic allodynia were tested on the day before surgery and on the 7th and 14th post-surgery days. Key markers of the nitrosative and oxidative stresses, in addition to markers of inflammation, were measured at day 14 post surgery. Histopathological examination and immunostaining of both synaptophysin and caspase-3 of sciatic nerve and brain stem were also performed. Results of this study showed that T. algeriensis extract suppresses UVA oxidative stress in HaCaT cells via activation of the Nrf-2 pathway. Both extracts attenuated hyperalgesia and allodynia at 7- and 14-days post-surgery with more prominent effects at day 14 of surgery. Their protective effects against neuropathic pain were mediated by inhibiting NOX-1, iNOS, by increasing the enzyme activity of catalase, and inhibition of inflammatory mediators, NF-κB, TNF-α, lipoxygenase, COX-2 enzymes, and PGE2. Furthermore, they improved deleterious structural changes of the brainstem and sciatic nerve. They also attenuated the increased caspase-3 and synaptophysin. The data indicate that both extracts have neuroprotective effects against chronic constriction injury-induced neuropathic pain. The observed protective effects are partially mediated through attenuation of oxidative and nitrosative stress and suppression of both neuroinflammation and neuronal apoptosis, suggesting substantial activities of both extracts in amelioration of painful peripheral neuropathy.

Element contents in different types of spices and aromatic herbs collected from Italy and Tunisia were evaluated. The aim was to evaluate: the possible differences and/or similarities among the analyzed samples; if it is possible related the samples to their geographical origins; the nutritional quality and the potential health risks. Potassium, Ca, Mg and Ni were low in laurel and rosemary; mint and thyme showed the highest Na and the lowest Se contents; arsenic and Cd levels were found highest in verbena which had also the lowest Hg content; lastly, black pepper had the highest Mn and the lowest Pb contents. Instead discrimination between Italian and Tunisian samples for each spices and aromatic herbs under analysis was achieved by PCA. Essential elements intake through samples was small. Levels of Pb in some samples exceed the maximum allowable level, but any safety risk for consumers is excluded., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The checkerboard method was used to study the potential interactions between eight essential oils (Basil, Cinnamon, Eucalyptus, Mandarin, Oregano, Peppermint, Tea tree, and Thyme) when used as antibacterial agents against Bacillus cereus LSPQ 2872 and Paenibacillus amylolyticus ATCC 9995. The minimum inhibitory concentration (MIC) of each essential oil (EO) and the fractional inhibitory concentration (FIC) index for the binary combinations of essential oils (EOs) were determined. According to FIC index values, some of the compound binary combinations showed an additive effect; however, Thyme/Tea tree and Cinnamon/Thyme EOs exhibited a synergistic effect against P. amylolyticus and B. cereus, respectively. Cinnamon/Thyme EOs mixture exhibited no interactive effect against P. amylolyticus, but a synergistic effect against B. cereus. The combination of Oregano/Thyme EOs displayed the best antibacterial activity and showed a synergistic effect against B. cereus and P. amylolyticus bacteria. The Oregano/Thyme EOs mixture has potential application in food preservation to reduce the contamination of B. cereus and P. amylolyticus., Competing Interests: Declaration of competing interest All authors of the manuscript state that there is no conflict of interest., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

1,8-Cineole is the main volatile produced by Thymus albicans Hoffmanns. & Link 1,8-cineole chemotype. To understand the contribution of distinct plant organs to the high 1,8-cineole production, trichome morphology and density, as well as emitted volatiles and transcriptional expression of the 1,8-cineole synthase (CIN) gene were determined separately for T. albicans leaves, bracts, calyx, corolla and inflorescences. Scanning electron microscopy (SEM) and stereoscope microscopy observations showed the highest peltate trichome density in leaves and bracts, significantly distinct from calyx and corolla. T. albicans volatiles were collected by solid phase micro extraction (SPME) and analyzed by gas chromatography-mass spectrometry (GC/MS) and by GC for component identification and quantification, respectively. Of the 23 components identified, 1,8-cineole was the dominant volatile (57-93 %) in all T. albicans plant organs. The relative amounts of emitted volatiles clearly separated vegetative from reproductive organs. Gene expression of CIN was assigned to all organs analyzed and was consistent with the relatively high emission of 1,8-cineole in leaves and bracts. Further studies will be required to analyze monoterpenoid biosynthesis by each type of glandular trichome., (© 2020 Wiley-VHCA AG, Zurich, Switzerland.)

The essential oils and antioxidant activity of four Thymus species were evaluated under five light spectra (namely, red, blue, red-blue, white, and greenhouse condition). The highest essential oil yield (4.17%) was observed under red light in T. migricus, while the lowest (1.05%) was observed in T. carmanicus under greenhouse conditions. Light quality also led to difference in essential oil constituents. The highest thymol (66%) was found in T. migricus exposed to blue light, while the least (1.69%) was observed in T. kotschyanus grown under red-blue light. The LED treatments did not induce any significant effect on carvacrol of Thymus species in comparison to the greenhouse condition. Finally, the analysis of variance indicates that the effect of light varied with the studied species. T. migricus performed the highest antioxidant capacity (IC 50  = 176.8 µg/mL) under blue light. Overall, essential oil components as well as antioxidants showed significant responses to light emitting diodes wavelengths., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Thyme ( Thymus spp .) volatiles predominantly consisting of monoterpenes and sesquiterpenes, serve as antimicrobial, antiseptic and antioxidant in phytomedicine. They also play a key role in plants as secondary metabolites via their potential role against herbivores, attracting pollinators and abiotic stress tolerance. Plant volatiles are affected by different environmental factors including drought. Here, the effect of prolonged water deficit stress on volatile composition was studied on the sensitive and tolerant thyme plant cultivars ( T. vulgaris Var. Wagner and T. vulgaris Var. Varico3, respectively). Volatile sampling along with morpho-physiological parameters such as soil moisture, water potential, shoot dry weight, photosynthetic rate and water content measurements were performed on one-month-old plants subsequent to water withholding at 4-day intervals until the plants wilted. The tolerant and sensitive plants had clearly different responses at physiological and volatile levels. The most stress-induced changes on the plants' physiological traits occurred in the photosynthetic rates, where the tolerant plants maintained their photosynthesis similar to the control ones until the 8 th day of the drought stress period. While the analysis of the volatile compounds (VOCs) of the sensitive thyme plants displayed the same pattern for almost all of them, in the tolerant plants, the comparison of the pattern of changes in the tolerant plants revealed that the changes could be classified into three separate groups. Our experimental and theoretical studies totally revealed that the most determinant compounds involved in drought stress adaptation included α-phellandrene, O-cymene, γ-terpinene and β-caryophyelene. Overall, it can be concluded that in the sensitive plants trade-off between growth and defense, the tolerant ones simultaneously activate their stress response mechanism and continue their growth.

In this research, metabolic profiling/pathways of Thymus vulgaris (thyme) plant were assessed during a water deficit stress using an FTICR mass spectrometry-based metabolomics strategy incorporating multivariate data analysis and bioinformatics techniques. Herein, differences of MS signals in specific time courses after water deficit stress and control cases without any timing period were distinguished significantly by common pattern recognition techniques, i.e., PCA, HCA-Heatmap, and PLS-DA. Subsequently, the results were compared with supervised Kohonen neural network (SKN) ones as a non-linear data visualization and capable mapping tool. The classification models showed excellent performance to predict the level of drought stress. By assessing variances contribution on the PCA-loadings of the MS data, the discriminant variables related to the most critical metabolites were identified and then confirmed by ANOVA. Indeed, FTICR MS-based multivariate analysis strategy could explore distinctive metabolites and metabolic pathways/profiles, grouped into three metabolism categories including amino acids, carbohydrates (i.e., galactose, glucose, fructose, sucrose, and mannose), and other metabolites (rosmarinic acid and citrate), to indicate biological mechanisms in response to drought stress for thyme. It was achieved and approved through the MS signals, genomics databases, and transcriptomics factors to interpret and predict the plant metabolic behavior. Eventually, a comprehensive pathway analysis was used to provide a pathway enrichment analysis and explore topological pathway characteristics dealing with the remarkable metabolites to demonstrate that galactose metabolism is the most significant pathway in the biological system of thyme., (Copyright © 2019 Elsevier B.V. All rights reserved.)

In this study, the fumigant antifungal activity of 10 Lamiaceae plant essential oils was evaluated against two phytopathogenic fungi, Raffaelea quercus-mongolicae, and Rhizoctonia solani. Among the tested essential oils, thyme white ( Thymus vulgaris ) and summer savory ( Satureja hortensis ) essential oils exhibited the strongest fumigant antifungal activity against the phytopathogenic fungi. We analyzed the chemical composition of two active essential oils and tested the fumigant antifungal activities of the identified compounds. Among the tested compounds, thymol and carvacrol had potent fumigant antifungal activity. We observed reactive oxygen species (ROS) generation in two fungi treated with thymol and carvacrol. Confocal laser scanning microscopy images of fungi stained with propidium iodide showed that thymol and carvacrol disrupted fungal cell membranes. Our results indicated that ROS generated by thymol and carvacrol damaged the cell membrane of R. querqus-mongolicae and R. solani , causing cell death.

The influence of homogenization conditions on selected essential oil (thyme, lemongrass, cinnamon, peppermint, and clove)-in-water nanoemulsion formation and stability was investigated. Physically stable essential oil nanoemulsions could be fabricated by a microfludizer under optimized processing conditions (10,000 psi and 2 passes). The chemical compositions of EOs was characterized using GC-MS. The antifungal activity and mycotoxin inhibitory activity of essential oils in both bulk and nanoemulsion forms were determined using two isolates of Fusarium graminearum. The major chemical components of essential oil had a remarkable impact on long term physical stability, antifungal activity, and inhibition of mycotoxin production. With regard to inhibition of mycotoxin production, the mycotoxin inhibitory activity of essential oils was enhanced considerably in nanoemulsion form, which was attributed to greater solubility of the essential oils. It was also noted that the same essential oils exhibited significant differences in inhibition of mycotoxin production in the two isolates of F. graminearum., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

During the last years, the food industry is working on the replacement of high energy methodologies with more sustainable techniques for the encapsulation of natural preservatives, in order to enhance their effectiveness as food additives. In the present study, nisin, an antimicrobial agent, was encapsulated in essential oil-containing microemulsions. More specifically, rosemary, thyme, oregano, and dittany essential oil-containing microemulsions were formulated to encapsulate nisin enhancing the system's overall antimicrobial activity. The systems were investigated for the interfacial properties and size of the surfactants' monolayer using electron paramagnetic resonance spectroscopy and dynamic light scattering. Subsequently, nisin-loaded microemulsions were tested for their antimicrobial activity against Lactococcus lactis, Staphylococcus aureus, Listeria monocytogenes, and Bacillus cereus, using the well diffusion assay. Finally, this technique was validated by a killing assay. Overall, this study provides important information on the antibacterial activity of nisin-loaded nano-carriers enhanced by essential oils, in relation to the microemulsions' structure., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Fluconazole (FLZ) application as a highly successful commercial antifungal azole agent to treat the fungal infections is limited due to emergence of FLZ-resistant candida. In this study, the potential of green synthesised silver nanoparticles (NPs) as an antifungal agent against Candida albicans fungal pathogen is investigated. The extract of ginger ( Zingiber officinale ) and thyme ( Thymus vulgaris ) plays as reducing agent, capping agent and antifungal agent. The UV-visible spectroscopy shows the peak of surface plasmon resonance of synthesised Ag NPs after a period of time. The synthesised Ag NPs are spherical, with average sizes of 12 and 18 nm based on ginger and thyme extract, respectively. Fourier transform infrared spectroscopy confirms the adsorption of the plant extract on the surface of the as-prepared Ag NPs. Based on the minimum inhibitory concentration (MIC) method against Candida albicans, the antifungal activity of as-prepared green synthesised Ag NPs shows higher inhibitory in comparison to FLZ. Finally, the Ag NPs synthesised via thyme extract shows no cytotoxicity with concentration below 3.5 ppm, which can be considered as an appropriate candidate instead of FLZ to treat the superficial fungal infections.

Thymus vulgaris L. (Lamiaceae), a well-known aromatic medicinal herb, has many important essential constituents in its oil, including γ-terpinene, carvacrol, thymol, and p-cymene. Gibberellins comprise hundreds of components, which regulate several various growths and underlying developmental processes, such as cell division and elongation, shoot elongation, seed germination, and gene expression. In this study, we investigated the influence of sprayed gibberellic acid (GA 3 ) treatments on the internode length, leaf morphology, length of new shoot, expression of monoterpene synthase genes and monoterpenes content during two plant growth stages. Our results showed that increasing of internode length was a clear effect of GA 3 that was varied with internode position. The results also showed that all internodes displayed a dramatic increase in the highest concentration of GA 3 . Also, the foliar application of GA 3 resulted in not only an increased expression level of monoterpene synthase genes, but also the improved production of a monoterpene, especially in the moderate concentration of GA 3 that they were up-regulated. In the lowest GA 3 concentrations, relative expression levels were similar or lower than the control plants and a notable downregulation in those genes was observed in the application of the highest concentration of GA 3 rather than the moderate concentrations. Overall, the expression of two out of five monoterpene synthase genes, TPS and CYP71D181, showed a correlation with the level of γ-terpinene and carvacrol, respectively, indicating that they are regulated at the transcriptional levels., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

Drought is one of the most important threats to plants and agriculture; therefore, understanding of the mechanism of drought tolerance is crucial for breeding of drought tolerant plants. Here, we assessed effects of four levels of drought (90%, 55%, 40% and 25% FC) on some physiological criteria and metabolite adjustment of two different drought-responsive thyme plants (Thymus vulgaris as drought sensitive and T. Kotschyanus as drought tolerant species), using 1 H-NMR. Among three physiological parameters and 18 identified metabolites, species × treatment effects were significant (P ≤ 0.01) for leaf temperature, acetic acid, citric acid, fumaric acid, malic acid, succinic acid, fructose, sucrose and serine. RWC, chlorophyll and carotenoids content, glucose, alanine and choline were affected by simple effects of species and treatment. Correlation analysis revealed that there is a different correlation between physiological parameters and metabolites in both species. This analysis also revealed that, by ignoring the correlation between malic acid and succinic acid in T. vulgaris, there was no significant correlation between TCA intermediate in both species. According to results, sugars, amino acid and energy metabolism were affected by drought and, among them, TCA intermediates had more alternation in two studied species so, this cycle and its intermediates probably have more prominent role than other identified metabolites in the induction of drought tolerance., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

Honey bees are highly prone to infectious diseases, causing colony losses in the worst case. However, they combat diseases through a combination of their innate immune system and social defence behaviours like foraging for health-enhancing plant products (e.g. nectar, pollen and resin). Plant secondary metabolites are not only highly active against bacteria and fungi, they might even enhance selective foraging and feeding decisions in the colony. Here, we tested six major plant terpenes and their corresponding acetates, characterizing six natural Thymus vulgaris chemotypes, for their antimicrobial activity on bacteria associated with European foulbrood. Comparison of the inhibitory activity revealed the highest activity for carvacrol and thymol whereas the acetates mostly did not inhibit bacterial growth. All terpenes and acetates are present in the nectar and pollen of thyme, with pollen containing concentrations higher by several orders of magnitude. The physiological response was tested on forager and freshly emerged bees by means of antennal electroantennography. Both responded much stronger to geraniol and trans-sabinene hydrate compared to carvacrol and thymol. In conclusion, bee-forageable thyme product terpenes (mainly from pollen) yield effective antibiotic activity by reducing the growth of bee disease-associated bacteria and can be detected with different response levels by the honey bees' antennae. This is a further step forward in understanding the complex pathogen-pollinator-plant network.

Thymus plants are marketed for diverse usages because of their pleasant odor, as well as high nutritional value and wealth of health-promoting phytochemicals. In this study, Thymus zygis , Thymus pulegioides , and Thymus fragrantissimus grown under organic cultivation regime were characterized regarding nutrients and phenolic compounds. In addition, the antioxidant and antibacterial properties of these species were screened. The plants were particularly notable for their high K/Na ratio, polyunsaturated fatty acids content and low omega-6/omega-3 fatty acids ratios, which are valuable features of a healthy diet. Caffeic acid and/or its derivatives, mainly rosmarinic acid and caffeoyl rosmarinic acid, represented the majority of the phenolic constituents of these plants, although they were less representative in T. pulegioides , which in turn was the richest in flavones. The latter species also exhibited the highest antioxidant capacity (DPPH ● EC 50 of 9.50 ± 1.98 μg/mL and reducing power EC 50 of 30.73 ± 1.48 μg/mL), while T. zygis was the most active towards Gram-positive and Gram-negative bacteria. Overall, the results suggest that the three thyme plants grown in organic farming are endowed with valuable metabolites that give them high commercial value for applications in different industries.

Thymus quinquecostatus Celak is a species of thyme in China and it used as condiment and herbal medicine for a long time. To set up the quality evaluation of T. quinquecostatus , the response surface methodology (RSM) based on its 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity was introduced to optimize the extraction condition, and the main indicator components were found through an UPLC-LTQ-Orbitrap MS n method. The ethanol concentration, solid-liquid ratio, and extraction time on optimum conditions were 42.32%, 1:17.51, and 1.8 h, respectively. 35 components having 12 phenolic acids and 23 flavonoids were unambiguously or tentatively identified both positive and negative modes to employ for the comprehensive analysis in the optimum anti-oxidative part. A simple, reliable, and sensitive HPLC method was performed for the multi-component quantitative analysis of T. quinquecostatus using six characteristic and principal phenolic acids and flavonoids as reference compounds. Furthermore, the chemometrics methods (principal components analysis (PCA) and hierarchical clustering analysis (HCA)) appraised the growing areas and harvest time of this herb closely relative to the quality-controlled. This study provided full-scale qualitative and quantitative information for the quality evaluation of T. quinquecostatus , which would be a valuable reference for further study and development of this herb and related laid the foundation of further study on its pharmacological efficacy.

Polyphenolic compounds are considered valuable secondary plant metabolites owing to the myriad of biological activities they exert. This study aimed to investigate the effect of applying various concentrations of the plant growth regulator, salicylic acid (SA), on Thymus vulgaris L. while subjecting the plant to decreasing amounts of irrigation water. The following parameters were monitored; total polyphenolic and flavonoid content, yield and composition of the essential oil, and antioxidant activity of the alcoholic extracts. Drought alone significantly (P < 0.05) increased the polyphenolic and flavonoid content, yield of the essential oil and antioxidant activity. The total flavonoid content in control plants was 6.1 ± 0.3 mg/gm dry weight calculated in terms of rutin equivalent. However, in drought stressed plants, (irrigated at 25% of the field capacity) sprayed with 3 mM SA, the flavonoid content increased to 32.1 ± 0.1 mg/gm dry weight calculated in terms of rutin equivalent. Moreover, the total phenolic content increased from 8.5 ± 0.3 to 68.5 ± 1.2 mg/gm dry weight calculated in terms of gallic acid in the same test plants. Radical scavenging activity, using DPPH assay, was measured for the different plant treatments. A decrease from 74.4 ± 0.4 μg/ml to 36.6 ± 0.9 μg/ml of IC 50 was recorded in the drought stressed plants (25% FC) sprayed with 3 mM SA compared with the control plants. The variability in polyphenolic composition between the control plants and plants with the highest total polyphenolic content was investigated by UPLC-ESI-MS/MS. Rosmarinic acid was detected as the major component in samples from both treatments, with a higher percentage observed upon subjecting the plant to the test conditions (25% FC and sprayed with 3 mM SA). The highest yield of the essential oil (1 ± 0.06 %v/w) was obtained from drought stressed plants (25% FC) sprayed with 2 mM SA. GC/MS analysis of oil samples revealed that the Thymol content increased with drought stress, while that of p-cymene decreased. However, an increase of p-cymene was witnessed as a result of SA spraying., (Copyright © 2017. Published by Elsevier Masson SAS.)

The essential oil of Thymus albicans Hoffmanns. & Link, a native shrub from the Iberian Peninsula, is mainly composed of monoterpenes. In this study, a 1,8-cineole synthase was isolated from the 1,8-cineole chemotype. A partial sequence that lacked the complete plastid transit peptide but contained an extended C-terminal when compared to other related terpene synthases was generated by PCR and Rapid Amplification of cDNA Ends (RACE). The predicted mature polypeptide was 593 amino acids in length and shared 78% and 77% sequence similarity with the homologue 1,8-cineole synthase from Rosmarinus officinalis and Salvia officinalis, respectively. The putative protein possessed the characteristic conserved motifs of plant monoterpene synthases including the RRx 8 W and DDxxD motifs and phylogenetic analysis indicated that the amplified 1,8-cineole synthase bears greater sequence similarity with other 1,8-cineole synthases from Lamiaceae family relative to the terpene synthases from the genus Thymus. Functional expression of the recombinant protein in Escherichia coli revealed that in the presence of geranyl diphosphate (GPP) 1,8-cineole was the major product but that its production was too low for robust quantification. Other minor conversion products included α-pinene, β-pinene, sabinene and β-myrcene suggesting the isolated 1,8-cineole synthase may be a multi-product enzyme. To our knowledge, this is the first report of a functionally characterized monoterpene synthase from Thymus albicans., (Copyright © 2017 Elsevier GmbH. All rights reserved.)

Phytogenic feed additives represent a potential alternative to antibiotics with attributed health and growth-promoting effects. Chickens supplemented with an essential oil blend, a Quillaja saponin blend, or a combination of both phytogenic preparations showed a comprehensively and significantly improved apparent ileal digestibility of crude protein and amino acids compared to control birds. Accordingly, holistic transcriptomic analyses of jejunum and liver samples indicated alterations of macromolecule transporters and processing pathways likely culminating in an increased uptake and metabolizing of carbohydrates and fatty acids. Complementary analyses in Caco-2 showed a significant increase in transporter recruitment to the membrane (SGLT1 and PEPT1) after addition of essential oils and saponins. Although the penetrance of effects differed for the used phytogenic feed additives, the results indicate for an overlapping mode of action including local effects at the intestinal border and systemic alterations of macronutrient metabolism resulting in an improved performance of broilers.

Thyme as a perennial herb has been recognized globally for its antimicrobial, antiseptic and spasmolytic effects. In this investigation, we have used non-targeted metabolite and volatile profiling combined with the morpho-physiological parameters in order to understand the responses at the metabolite and physiological level in drought sensitive and tolerant thyme plant populations. The results at the metabolic level identified the significantly affected metabolites. Significant metabolites belonging to different chemical classes consisting amino acids, carbohydrates, organic acids and lipids have been compared in tolerant and sensitive plants. These compounds may take a role through mechanisms including osmotic adjustment, ROS scavenging, cellular components protection and membrane lipid changes, hormone inductions in which the key metabolites were proline, betain, mannitol, sorbitol, ascorbate, jasmonate, unsaturated fatty acids and tocopherol. Regarding with volatile profiling, sensitive plants showed an increased-then-decreased trend at major terpenes apart from alpha-cubebene and germacrene-D. In contrast, tolerant populations had unchanged terpenes during the water stress period with an elevation at last day. These results suggesting that the two populations are employing different strategies. The combination of metabolite profiling and physiological parameters assisted to understand precisely the mechanisms of plant response at volatile metabolome level., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Thyme (Thymus vulgaris L.) is known to produce a variety of phenolic monoterpenes such as thymol and carvacrol. Thymol and carvacrol are health-promoting, biocide and antitoxin compounds and have been considered as the main constituents of essential oils in T. vulgaris. To improve our understanding of the regulation of monoterpene biosynthesis in thyme, the expression of genes related to thymol and carvacrol biosynthesis in different tissues and in response to abiotic elicitors was analyzed. Methyl jasmonate (MeJA), salicylic acid (SA), trans-cinnamic acid (tCA) and UV-C irradiation were applied to T. vulgare leaves and transcript levels of early (DXR) and late (TvTPS1, CYP71D178 and CYP71D180) biosynthetic genes of thymol and carvacrol were measured. The results showed that early step and late step genes in thymol/carvacrol biosynthesis are differentially regulated. DXR was not found to be exclusively expressed in glandular trichomes; in contrast, biosynthetic genes including γ-terpinene synthase (TvTPS1) and two cytochrome P450s, CYP71D178 and CYP71D180, were preferentially expressed in glandular secretory trichomes. The high expression of late biosynthetic genes in glandular trichomes, which also contain the highest concentration of thymol and carvacrol, suggests that glandular trichomes are the structure in which thymol/carvacrol biosynthesis and accumulation occur. Our results indicate that in addition to abiotic elicitors, developmental and spatial factors also play a key role in the biosynthesis of thymol and carvacrol, most likely relating to glandular trichome density and/or activity. Hence optimization of these factors could be considered as a useful strategy to achieve high yield of valuable compounds in T. vulgare or other closely related plant species., (Copyright © 2017 Elsevier Masson SAS. All rights reserved.)

Thymus munbyanus subsp. coloratus (Lamiaceae) is a small shrub endemic to Algeria and Morocco where is found in lawns, rockeries and mountainous regions. From a phytochemical point of view this taxon has never been characterized. In this work we have analysed the chemical compositions of the essential oils obtained from inflorescences and vegetative parts by GC/MS. A new chemotype, i.e. borneol-chemotype, was characterized for the first time in the species. Furthermore, we assessed the biological activities of essential oils, namely the antioxidant, antimicrobial and cytotoxicity on tumor cells that were evaluated by the DPPH, ABTS, and FRAP, disc diffusion, and MTT methods, respectively. Biological assays highlighted a moderate inhibitory effect on Staphylococcus aureus, Escherichia coli and Candida albicans (inhibition zone diameter in the range 9 - 10 mm), and noteworthy cytotoxicity on A375 human melanoma cells (IC 50 of 46.95 μg/ml)., (© 2017 Wiley-VHCA AG, Zurich, Switzerland.)

Consumers' demand for ready-to-eat (RTE) turkey meat is attributed to its convenience and healthy properties. However, as cooked meat product it is subjected to post-process contamination, thus allowing presence and growth of microbial pathogens, such as Salmonella spp.. The aim of this study was to include a natural antimicrobial, thyme essential oil (TEO), on RTE turkey products in order to evaluate its effectiveness throughout the shelf life. To do so, the effect of four different formulations of cooked RTE turkey products on Salmonella Enteritidis behaviour was investigated. Products' slices were surface inoculated with S. Enteritidis (ca. 4 to 5logcfu/g), subsequently stored at 10 and 25°C and microbiologically analysed during 18 and 12days, respectively. Predictive microbiology models fitted to count data were used to evaluate microbial behaviour. Results showed that S. Enteritidis behaviour on RTE turkey products slices during storage was strongly dependent on temperature. The pathogen was able to grow on slices at all tested conditions during storage at 25°C and no statistical differences were detected (p>0.05) between growth parameters. At 10°C, different behaviour patterns were observed. The application of TEO led to higher Salmonella inactivation rates on a product exempt of chemical preservatives. The addition of this novel antimicrobial on meat products or its incorporation on meat active packaging systems as a part of hurdle technology could increase RTE turkey products safety while satisfying the demand of more natural foods., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The aim of this study was to improve the oxidative stability of soybean oil by using aromatic plants. Soybean oil flavored with rosemary (ROS) and soybean oil flavored with thyme (THY) were subjected to heating for 24h at 180°C. The samples were analyzed every 6h for their total polar compounds, anisidine values, oxidative stability and polyphenols content. The tocopherols content was determined and volatile compounds were also analyzed. After 24h of heating, the incorporation of these plants using a maceration process reduced the polar compounds by 69% and 71% respectively, in ROS and THY compared to the control. Until 6h of heating, the ROS kept the greatest oxidative stability. The use of the two extracts preserves approximately 50% of the total tocopherols content until 18h for the rosemary and 24h for the thyme flavored oils. Volatile compounds known for their antioxidant activity were also detected in the formulated oils. Aromatic plants added to the soybean oil improved the overall acceptability of potato crisps (p<0.05) until the fifteenth frying., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

A pot experiment was carried out with lemon balm (Melissa officinalis L.) and thyme (Thymus vulgaris L.). Different water supply was applied: 25%, 40% and 70% saturation of soil water capacity (SWC). Morphological traits, biomass and phenolic type active ingredients were investigated. Among the two species, main differences were registered in biomass and TPC. Lower SWC resulted in reduced biomass production of lemon balm, while the applied stress treatments did not effect the biomass of thyme. In lemon balm, highest TPC contents were measured in control plants both in shoots and roots but in thyme, the shoots showed a significantly increased TPC at the 25% SWC conditions. Neither the content of total flavonoids nor that of the rosmarinic acid was affected by the treatments. The antioxidant capacity proved to be in tight connection with the TPC in both species (r = 0.766-0.883). The rosmarinic acid content of lemon balm plants contributed to the antioxidant capacity, as well (r = 0.679-0.869).

High blood pressure is the most powerful contributor to the cardiovascular morbidity and mortality, and inverse correlation between consumption of polyphenol-rich foods or beverages and incidence of cardiovascular diseases gains more importance. Reactive oxygen species plays an important role in the development of hypertension. We found that wild thyme (a spice plant, rich in polyphenolic compounds) induced a significant decrease of blood pressure and vascular resistance in hypertensive rats. The inverse correlation between vascular resistance and plasma heme oxygenase-1 suggests that endogenous vasodilator carbon monoxide generated by heme oxidation could account for this normalization of blood pressure. Next product of heme oxidation, bilirubin (a chain-breaking antioxidant that acts as a lipid peroxyl radical scavenger), becomes significantly increased after wild thyme treatment and induces the reduction of plasma lipid peroxidation in hypertensive, but not in normotensive rats. The obtained results promote wild thyme as useful supplement for cardiovascular interventions.

There is a growing interest in the use of naturally-occurring antimicrobial agents such as plant essential oils (EOs) to inhibit the growth of hazardous and spoilage microorganisms in foods. Gaseous EOs (EO gases) have many potential applications in the food industry, including use as antimicrobial agents in food packaging materials and sanitizing agents for foods and food-contact surfaces, and in food processing environments. Despite the potentially beneficial applications of EO gases, there is no standard method to evaluate their antimicrobial activities. Thus, the present study was aimed at developing an experimental apparatus and protocol to determine the minimal inhibitory concentration (MIC) and minimal lethal concentration (MLC) of EO gases against microorganisms. A sealed experimental apparatus was constructed for simultaneous evaluation of antimicrobial activities of EO gases at different concentrations without creating concentration gradients. A differential medium was then evaluated in which a color change allowed for the determination of growth of glucose-fermenting microorganisms. Lastly, an experimental protocol for the assessment of MIC and MLC values of EO gases was developed, and these values were determined for 31 EO gases against Escherichia coli O157:H7 as a model bacterium. Results showed that cinnamon bark EO gas had the lowest MIC (0.0391 μl/ml), followed by thyme-thymol EO gas (0.0781 μl/ml), oregano EO gas (0.3125 μl/ml), peppermint EO gas (0.6250 μl/ml), and thyme-linalool EO gas (0.6250 μl/ml). The order of the MLC values of the EO gases against the E. coli O157:H7 was thyme-thymol (0.0781 μl/ml)

Purpose: The purpose of this study was to evaluate the effects of essential oil on oxidative stress, immunity, and skin condition in atopic dermatitis (AD) induced mice., Methods: This study was a 3×3 factorial design. Factors were oil type (Lavender, Thyme, and 2:1 mixture of lavender and thyme oil [blending oil]) and treatment period (0 day, 7 days, and 21 days). The samples were 45 mice with AD and randomly assigned to nine groups of five mice per group. The dependent variables such as superoxide radical, IgE, degranulated mast cells, and epidermal thickness were measured. Data were collected from February to April in 2014. Descriptive statistics, One-way ANOVA, Two-way ANOVA, and Tukey's HSD test were performed using the SPSS WIN 20.0 program., Results: Dependent variables were not statistically significantly different by the three oil types (p>.05). Essential oils such as lavender, thyme, and blending oil were all effective in reducing AD symptoms and especially 2:1 blending oil were most effective. There were statistically significant differences by the three treatment periods in all dependent variables (p<.001). There were statistically significant interactions between oil types and treatment periods in all dependent variables (p<.01). For decreasing superoxide radical, degranulated mast cells, and epidermal thickness, 2:1 mixed oil should be applied for at least 21 days. Otherwise to reduce IgE, 2:1 mixed oil should be used for at least 7 days., Conclusion: These findings provide bases for developing effective interventions for AD patients to manage their AD symptoms.

The antifungal and antiaflatoxigenic properties of Thymus vulgaris essential oil (TEO) were evaluated upon Aspergillus flavus "in vitro". Suspension containing 10(6) of A. flavus were cultivated with TEO in concentrations ranging from 50 to 500 μg/mL. TEO reached minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) at 250 μg/mL. Inhibition of ergosterol biosynthesis was detected at a concentration of 100 μg/mL of TEO. Morphological evaluation performed by both light microscopy and scanning electron microscopy showed that antifungal activity of TEO could be detected starting at a concentration of 50 μg/mL and the fungicide effect at a concentration of 250 μg/mL. TEO completely inhibited production of both B1 and B2 aflatoxins (AFB1 and AFB2) at a concentration of 150 μg/mL. This way, fungal biomass development and aflatoxin production were dependent on TEO concentration. Therefore, TEO was capable of controlling the growth of A. flavus and its production of aflatoxins., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The gut metabolism of four thyme phenolics (monoterpenes thymol and carvacrol, rosmarinic acid, and eriodictyol) was evaluated in vitro. After the in vitro transformations of the individual phenols had been studied, the presence of their microbial metabolites was investigated in human feces collected before and after a sustained intake (3 weeks) of 25 mL/day of a thyme phenol-enriched olive oil. Results of in vitro fermentation showed low degradation of thymol and carvacrol. By contrast, large catabolism was noted when rosmarinic acid and eriodictyol were fermented, yielding hydroxyphenylpropionic acid as the main metabolite. In accordance with these results, after the in vivo intervention with thyme phenol-enriched olive oil, an increase in the concentration of hydroxyphenylpropionic and phenylpropionic acids was observed in human feces, confirming the effective in vivo microbial degradation of rosmarinic acid and eriodictyol. Carvacrol was detected in fecal samples at trace levels, suggesting that monoterpenes are well absorbed in the upper part of the gastrointestinal tract.

Thymus caramanicus is an endemic species grown in Iran with interesting pharmacological and biological properties. In the present work, essential oil compositions and inter-simple sequences repeat (ISSR) markers were used to estimate the relationships among and within seven populations of T. caramanicus, belonging to three provinces in Iran. The studied individuals were distinguished on the basis of ISSR markers and constituents of essential oil. A total of 127 band positions were produced by 12 ISSR primers, of which 105 were found polymorphic with 82.68% polymorphism. Genetic similarity values among individuals ranged between 0.15 and 0.82 which was indicative of a high level of genetic variation. On the basis of their genetic similarities, ISSR analysis allowed to group the samples into two main clusters. One of these included populations originated from Kerman and Isfahan provinces, and the other cluster consists of populations from Semnan province. Chemical compounds of essential oils were found variable in the various individuals and all samples were principally composed of phenolic constituents (carvacrol and/or thymol). As a consequence, the plants were classified into two major chemotypes including carvacrol and thymol/carvacrol. A relationship between genetic and chemical variability and geographic distribution has been observed in studied populations of T. caramanicus.

Olive oils flavoured with edible herbs have grown in popularity because of their added value and potential health benefits. However, the combined presence of different phytochemicals from olive oil and herbs requires study of their possible interactions during intestinal transport and metabolism. The aim of this study was firstly to evaluate the effect on bioaccessibility of the co-occurring bioactive compounds from olive oil and thyme through an in vitro digestion model of three extracts: olive extract (OE), thyme extract (TE) and a combination of both (OTE). The bioaccessible fractions were exposed to Caco-2 and HepG-2 cell models, as well as to a co-culture of both of these. Results indicated that the bioaccessibility of hydroxytyrosol was enhanced when OTE was digested. After Caco-2 cells exposure, no significant differences were observed in hydroxytyrosol transport, whereas the main flavonoids from thyme seemed to undergo an enhanced basolateral permeation when both phenolic sources where exposed., (Copyright © 2013 Elsevier Ltd. All rights reserved.)