This study investigatedthe effects of thymol (TH) adaptation onbiofilm formation and the metabolic profile of the multiresistantslaughterhouse isolate Pseudomonas psychrophila M33T02.2. After exposure to increasing sublethal concentrationsof thymol, the adapted P. psychrophila M33T02.2 showed decreased biofilm-forming capacity, reduced swarmingmotility, and lower rhamnolipid production compared to the wild-typestrain. Confocal microscopy further showed that the biofilms developedby the adapted strain were less homogeneous, confirming their inabilityto develop well-structured biofilms. To further understand these changesat the metabolic level, high-performance liquid chromatography (UHPLC-Orbitrap-MS/MS)identified redox metabolism intermediates and energy balance-relatedmetabolites as most important variables. 20 metabolites were underexpressedfor the TH-adapted strain, including glutathione disulfide, guanosinediphosphate, and flavin adenine mono- and di-nucleotide, among others.Therefore, we conclude that repeated exposure to TH prevents the emergenceof resistance mechanisms associated with biofilm formation, actingat the level of redox state and energy imbalance.

Synergistic effects of antimicrobial peptide KRR-N1-5W6L and thymol on the inactivation of methicillin-resistant Staphylococcus aureus in chilled beef.

Beeswax-enriched tricalcium phosphate/hydroxyapatite/sodium alginate/thymol 3D-printed scaffolds for application in bone tissue engineering.

ABSTRACT In this study, carvacrol (CA)/thymol (TH) nanoemulsions (CT NEs) were first prepared by the ultrasonic emulsification method, and then the effects of various nanoemulsion concentrations on the physicochemical properties and biological activities of chitosan/pullulan (CS/PU) nanocomposite films were systematically evaluated. The CT NEs exhibited good stability with a high encapsulation efficiency of 90.29% ± 0.95%, and they were well integrated in the CS/PU matrix as confirmed by the microstructure characterization. The performances of the nanocomposite films were significantly enhanced by CT NEs, including the improvements of mechanical properties and barrier properties. The CS/PU‐CT films exerted notable antioxidant and antibacterial activities. In the end, the CS/PU‐3CT film (with 3.0% w/v CA and TH), with the best antioxidant and antibacterial performance, was applied to the preservation of pork, and its shelf life was extended for 10 days compared to the CK group. The film loaded with 3CT NEs exhibits outstanding performance and offers significant prospects for use in food packaging.

Trans-ferulic acid (TFA), a phenolic acid abundant in fruits and cereals, and thymol (THY), a monoterpenoid phenol from thyme oils, are bioactive natural compounds with notable neuropharmacological potential. This study evaluated the combined anxiolytic effects of THY and TFA in Swiss albino mice, focusing on GABAergic mechanisms and pharmacokinetic profiles through in vivo and in silico methods. Mice received vehicle, diazepam (DZP, 2mg/kg), TFA (50mg/kg), THY (50mg/kg), or their combination (THY + TFA; 50 + 50mg/kg, p.o.). Behavioral assessments in open-field, swing, hole-cross, and light-dark tests indicated that the combined treatment produced more pronounced effects, markedly reducing locomotor and exploratory activity (e.g., square crosses: 25.8 ± 3.7 vs. 94.6 ± 6.2 in control; hole crosses: 3.4 ± 1.8 vs. 15.6 ± 1.9) and increasing time in the light chamber, consistent with anxiolytic-like activity comparable to DZP. Molecular docking targeted gamma-aminobutyric acid A (GABAA) receptor subunits (α2, α3), and ADME/toxicity profiles were analyzed in silico. Docking analysis revealed moderate binding affinities (BAs) to GABAA receptor subunits AF-P26048-F1 (α2 subunit) (- 5.8kcal/mol) and 8G4X (α3 subunit) (- 5.3kcal/mol) for TFA and α2 (- 4.7kcal/mol) and α3 (- 5.6kcal/mol) for THY, relative to DZP (- 6.9kcal/mol). These findings suggest that the THY + TFA combination elicits enhanced GABAergic modulation, offering a potential natural therapeutic strategy for anxiety management with reduced toxicity risk. However, formal interaction analyses (e.g., isobologram or two-way ANOVA) are warranted to confirm the combining effect.

Species diversity, benzimidazole resistance, and phytochemical synergy in gastrointestinal nematode nemabiomes from sheep farms in Portugal.

The quorum sensing (QS) system and cell-to-cell communication have had a significant impact on biofilm formation and virulence factor increase in Pseudomonas aeruginosa (P. aeruginosa), making this opportunistic pathogen a global concern and potentially life-threatening agent. The present study aimed to create an innovative pharmaceutical bio-nanocomposite (BNC) comprising thymol (THY) and gallic acid (GA) based on β-cyclodextrin (β-CD), which was used to investigate the release kinetics of active compounds, the level of cytotoxicity, antibacterial and anti-biofilm potential, and measuring the expression of genes effective in QS in the strain PAO1 pays P. aeruginosa. Based on this, physicochemical characteristics of the synthesized BNC were determined using Fourier transform infrared spectroscopy analysis (FTIR), UV-vis measurement, dynamic light scattering (DLS), zeta potential, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The BNC’s antibacterial and anti-biofilm capabilities were assessed using the PAO1 reference strain of P. aeruginosa and the expression level of QS-effective genes (rhlI, rhlR, lasI, and lasR) in bacteria was also evaluated in the presence of the synthesized BNC. The results of FTIR spectroscopy show the formation of intramolecular hydrogen bonds between THY, GA and β-CD. Absorption peaks of the UV-vis spectroscopy spectrum of the synthesized BNC at wavelengths of 217 and 272 nm are confirmed successful encapsulation of the THY and GA into the β-CD. The maximum size of the synthesized BNC was recorded as 356.3 nm with a polydispersity index (PDI) of 0.816. SEM and TEM micrographs show the presence of THY/GA active compounds in the pores in β-CD and the formation of a dense polymer network. After 360 minutes of release kinetics, more than 70% of the complex’s active chemicals had been released. The biological complex’s low toxicity is indicated by average cell survival of more than 65% and the ability to preserve the spindle shape of normal fibroblast cells at high concentrations. The PAO1 strain has minimum inhibitory concentrations (MIC) of 323 and 199.6 μg/mL for minimal biofilm inhibition concentration 50% (MBIC50). The decrease in rhlI and rhlR gene expression relative to the control group (without treatment) suggests that the active chemicals released from the biological complex interact and disrupt the QS pathway. Overall, the synthesized pharmaceutical complex has promise as a clever and effective option for future research and practical advances, as well as the development of complementary therapies.

This study aimed to investigate the effects of active clay nanocomposite packaging films based on linear low-density polyethylene (LLDPE) containing eugenol (EUG)/thymol (THY) or carvacrol (CRV) loaded layered montmorillonite (MMT) or tubular halloysite (HT) nanoclays on the natural microbial load (total mesophilic aerobic bacteria (TMAB), total coliforms (TC), total lactic acid bacteria (LAB), and total yeast-mold (TYM)) of kashar cheese, as well as on the presence/growth of Listeria monocytogenes, Staphylococcus aureus, and Aspergillus niger, along with pH values. The results showed that when prioritizing the packaging materials with the greatest impact on the microbial populations of kashar cheese, the order observed was as follows: THY-HNT for TMAB; EUG-MMT, CRV-HNT and THY-HNT for LAB; and CRV-HNT and THY-HNT for TYM. Moreover, S. aureus was not found in any of the kashar cheese samples. Furthermore, CRV-HNT and EUG-MMT films exhibited fungicidal properties throughout storage, leading to a decrease in mold spore counts, whereas THY-HNT films showed fungistatic effects. Additionally, CRV-HNT and THY-MMT packaging samples effectively limited growth of L. monocytogenes. As well, the pH values (5.15-5.52) of the cheese samples fluctuated during the storage period except for the control samples. Overall, active nanocomposite films had the potential to extend the shelf life of kashar cheese.

ABSTRACTNausea and vomiting are common and distressing responses to toxins, chemotherapy, and gastrointestinal disturbances. Although antiemetic medications are available, their adverse effects make safer substitutes necessary. In our study, Thymol (THY), a phenolic monoterpene from essential oils, was evaluated for its antiemetic potential using in vivo and in silico methods. In the in vivo study, emesis was induced in 2‐day‐old chicks by oral administration of copper sulfate pentahydrate (50 mg/kg). THY was administered orally at doses of 10, 20, and 40 mg/kg, alone or in combination with standard antiemetics ondansetron (ODN), domperidone (DPD), hyoscine butyl bromide (HYS), and promethazine hydrochloride (PRO). The 20 mg/kg dose (THY‐20) showed the highest efficacy, significantly (p < 0.0001) reducing the number of retches (24.6 ± 2.7; 67.2% reduction) and increasing latency to first retch (52.6 ± 4.2 s; 77.18% increase) compared to the negative control (NC). The THY + ODN combination further enhanced effects (68.53% retch reduction). Molecular docking showed strong binding of THY to 5‐HT3A (−6.4 kcal/mol), D2 (−7.1 kcal/mol), M3 (−6.2 kcal/mol), and H1 (−7.1 kcal/mol) receptors, comparable to standard drugs. Absorption, distribution, metabolism, excretion, and toxicity (ADMET) profiling revealed THY's compliance with Lipinski's Rule, high gastrointestinal absorption, blood–brain barrier permeability, and low toxicity risk. The multi‐target binding profile and synergistic potential of THY suggest its promise as a natural, broad‐spectrum antiemetic. Further receptor‐specific studies and trials in chemotherapy‐induced emesis models are recommended to validate its clinical potential.

ABSTRACTRecently, deep eutectic solvents (DESs) have become known as solvents that have highly tunable properties and are eco‐friendly. Several terpenes, such as thymol and menthol, have resonance effects, which subsequently lead to their polar asymmetry, and this factor causes the majority of V‐type DESs to be investigated based on these terpenes. An External Electric Field (EEF) affects the intermolecular interactions and the electronic state energies of DESs. The ubiquitous presence of electric fields in systems plays an important role in functions. For this purpose, in the present study, MD simulations have been performed to design and describe the structural properties and gain deeper insights into the behavior of DES consisting of Menthol (MEN) and Thymol (THY) in molar ratios (1:1) in the absence (T = 298, 350, and 398.15 K) and presence of an EEF with strengths of 0.217, 0.434, 0.651, and 0.868 (V/Å) at a temperature of 398.15 K and atmospheric pressure. This is the first time that the structural and dynamical characteristics of DES‐based terpenes under EEFs have been investigated. The response of the eutectic mixture to the external fields is analyzed using the dipolar arrangements, the intermolecular hydrogen bond network, and the relative orientations of the nearest neighbors. The simulation results showed that as the external electric field increased from 0.217 to 0.651 (V/Å), the distribution of solvent species around each other increased. The results indicate that with increasing temperature, the hydrogen bond network between components HBA and HBD was disturbed at high temperatures.

The use of natural products as antibiotic adjuvants to enhance efficacy and mitigate resistance is increasingly recognized as a promising strategy. This study explored five novel synergistic antimicrobial combinations (SACs) of carvacrol (CARV) and three already identified SACs of thymol (THY) with chloramphenicol, gentamicin, and streptomycin against Staphylococcus aureus and Acinetobacter baumannii, critical WHO-listed pathogens, and investigated their mechanisms of action and resistance-prevention capabilities. Despite being isomers, CARV and THY exhibited distinct synergistic effects and fractional inhibitory concentration index (FICI) values depending on the antibiotic and bacterial species. The SACs significantly reduced the required antibiotic dose by 4- to 16-fold, with FICI values ranging from 0.25 to 0.5. Growth kinetics revealed that SACs completely inhibited planktonic bacterial growth, outperforming antibiotics alone. Additionally, the SACs demonstrated efficacy in both inhibiting and eradicating biofilms of S. aureus and A. baumannii. Resistance development studies highlighted that neither THY nor CARV induced resistance in these pathogens. Moreover, SACs combining aminoglycosides with THY reduced the emergence of resistance in A. baumannii by up to 32-fold. In S. aureus, THY mitigated gentamicin resistance by 16-fold. CARV exhibited similar, albeit slightly less potent, effects.Mechanistic investigations revealed that THY and CARV exert antimicrobial action by multiple mechanisms, including bacterial membrane depolarization and disruption, efflux pump inhibition, disrupting ATP metabolism and mitigating oxidative stress induced by antibiotics. These findings highlight the potential of SACs to enhance antibiotic efficacy while preventing resistance, positioning them as strong candidates for innovative antimicrobial therapies against multidrug-resistant pathogens.Graphical

Metal-organic frameworks (MOFs) are a class of coordination polymers known for their intrinsic porosity. This property, combined with their ability to exhibit some degree of flexibility, makes them ideal materials for various innovative applications, like the inclusion and controlled release of active compounds for active packaging and agricultural applications. This study aimed to characterize the uptake and release behavior of an equimolar mixture of two phenolic essential oils (EOs), that is, carvacrol (CAR) and thymol (THY), encapsulated in two microporous Zn-based MOFs with different entanglement motifs: PUM168 is threefold interpenetrated and flexible, while PUM210 is fourfold polycatenated and rigid (PUM = Parma University Materials). The guest uptake was calculated using Nuclear Magnetic Resonance (1H NMR) spectroscopy and thermal gravimetric analysis (TGA) analysis, whereas the release profiles were investigated via headspace gas chromatography-mass spectrometry (HS-GC-MS) (Headspace-Gas Chromatography coupled with Mass-Spectrometry) analysis over a period of 15 days at room temperature. The monitoring revealed the superior ability of the flexible PUM168 for a stable and prolonged release of the volatile components, while the release from the rigid PUM210 was completed within 24 hours.

Enzyme-responsive bacteriostatic fibers loaded with thymol for chilled mutton preservation.

The construction of multipurpose particles with functional coatings of varying structure and composition provides the opportunity to modify their physicochemical and biological characteristics. In accordance with the aforementioned, new polyelectrolytes (PEs) decorated with an antimicrobial function (PEs-DAF) were designed and prepared to apply them as building blocks of a various carrier systems. A series of hydrogel nano- and microparticles were developed and coated with an outer PE shell with antimicrobial functionality. To this end, poly-(acrylic acid) (PAA) was grafted with different degrees of substitution (DS) of antimicrobial essential oils such as thymol (THY), menthol (MEN), and carvacrol (CAR) (abbreviated as PAA-X-DS% (X = THY, MEN, CAR; DS = 5,15)) using Steglich esterification under mild reaction conditions. Their structures were confirmed by 1H NMR and FTIR spectroscopy. The particles' morphology and mean diameter were determined by dynamic light scattering (DLS), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and atomic force microscopy (AFM). The physicochemical properties of the novel functional coatings were characterized using quartz crystal microbalance with a dissipation (QCM-D) analysis and spectroscopic ellipsometry. The antimicrobial properties of the functionalized PAA and the alginate microgel particles decorated with these PEs were evaluated against Staphylococcus aureus and Escherichia coli using the agar disc diffusion assay and minimal inhibitory concentration evaluation. The particles exhibited satisfactory antimicrobial activity, and some examples showed higher bioactivity than the functionalized PAAs. Moreover, the designed systems were loaded with resveratrol (RES), a model chemotherapeutic substance, to assess their potential applicability as drug carriers. The analysis proved the effective RES encapsulation and its release in a controlled manner depending on the coating properties. The results found in our study indicate potential therapeutic applications of the new antimicrobial-decorated carrier systems in the treatment of multidrug-resistant pathogenic infections.

Poultry housing includes unavoidable stressors that impair birds' welfare and health. Global policies are banning antimicrobial growth promoters mainly due to antimicrobial resistance. Antioxidants modulate immunity by reducing oxidative stress, which impairs immune function. Thymol (THY), tocopherol (TOC) and ascorbyl palmitate (AP) have been studied for their antioxidant, antimicrobial and anxiety/fear-reducing properties. However, their effects on immune function and stress responses in broilers require further investigation. This study assesses whether dietary THY or a TOC and AP mix modulates immune and chronic stress-related responses in broilers. Cobb-500 chicks (n=960) were assigned to one of 6 dietary treatments: (1) Basal (control), (2) Promotor (Basal+flavomycin), (3) BHT (Basal+butylated hydroxytoluene), (4) Promotor-BHT (Basal+flavomycin+BHT), (5) THY (Basal+thymol) and (6) TOC-AP (Basal+tocopherol+AP). Immune and stress parameters were evaluated, including inflammatory response to phytohemagglutinin-P (PHA-P), antibody production against sheep red blood cells (SRBC) and heterophil-to-lymphocyte (H/L) ratio. Supplementation did not affect the PHA-P inflammatory response (p=0.72) but influenced SRBC antibody production and H/L ratio (p<0.003). Broilers supplemented with THY or TOC-AP exhibited significantly lower antibody responses, potentially avoiding an energy-demanding acquired immune activation. Additionally, both groups showed significantly lower H/L ratios, suggesting that these supplements may help mitigate physiological stress induced by routine husbandry practices. These findings provide evidence that THY and TOC-AP may serve as natural alternative to synthetic additives for improving welfare and mitigating stress-induced immune imbalance under commercial rearing conditions.

Effect of bovine bone gelatin/sodium carboxymethyl-cellulose nanoemulsion loaded with thymol on chilled beef preservation.

Abstract Thymoquinone (TQ), a compound with diverse pharmacological benefits, faces limited availability from natural sources. In this study, the design of a highly efficient electrocatalyst, Pt clusters loaded on CeO2 with rich oxygen vacancies (Vo‐Pt/CeO2), for the electrocatalytic oxidation of thymol (TY) or carvacrol to produce TQ is reported. The introduction of oxygen vacancies to the surface of Vo‐Pt/CeO2 significantly reduces the adsorption energy of TQ, facilitating its desorption and preventing product accumulation and catalyst poisoning. As a result, Vo‐Pt/CeO2 demonstrates exceptional electrocatalytic performance, achieving ≈100% selectivity and up to 98% yield of TQ under 1.2 VRHE. In situ, Raman spectroscopy indicates that the electrocatalytic oxidation mechanism of TY involves water attacking the TY intermediate to form thymohydroquinone, which is subsequently oxidized to TQ. Overall, this work provides novel insights into the catalyst design and presents a new strategy for sustainable TQ synthesis.

Tuberculosis (TB) remains a persistent global health challenge, with an increasing incidence of cases and limitations in current treatment strategies. Traditional therapy involves long drug treatments that can cause side effects and lead to drug-resistant strains, making treatment less effective. This study aimed to explore the therapeutic potential of a novel nanoparticle-based delivery system for Thymol (THY), a natural antibacterial bioactive molecule, to combat Mycobacterium smegmatis, a model organism for Mycobacterium tuberculosis. A nanoparticle-based delivery system was developed using biocompatible Thymolconjugated Chitosan Zinc Ferrite Nanoparticles (THY-CH-ZnFe2O4 NPs). The nanoconjugates were characterized for their morphological and chemical properties. The characterization of synthesised nanoparticles showed THY-CH-ZnFe2O4 NPs to exhibit enhanced biocompatibility and antibacterial activity against M. smegmatis compared to THY alone. The nanoconjugates induced Reactive Oxygen Species (ROS)-mediated damage to the bacterial cell membrane, effectively inhibiting bacterial replication, dormancy, and biofilm formation. Additionally, the nanoconjugates demonstrated low cytotoxicity towards the human kidney cell line. The study's findings highlighted a new direction for developing nanoparticle-based antimycobacterial agents with a wide application in treating TB and other bacterial diseases. The THY-CH-ZnFe2O4 NPs show promise as a safe and effective therapeutic agent, offering a potential solution to the limitations of current TB treatment strategies.

Effect of different modified potato starch on starch-based films containing ZIF-67@thymol.

Monoterpene phenols have antibacterial, antioxidant, anticancer, antihypertensive, and antidiabetic properties, but their natural volatility and hydrophobicity limit their biomedical applications. In this work, we developed an oil-in-water nanoemulsion (NE) stabilized by proteins (i.e., sodium caseinate, NaCas) and polyphenols (i.e., tannic acid, TA) using phacoemulsification, where thymol (TH) was dissolved in carvacrol (CA) and the mixture was used as the dispersed phase. The NE avoids the use of traditional surfactants as the stabilizer. Amphiphilic NaCas could stabilize the NE, followed by further cross-linking of NaCas using TA. These antimicrobial agents of monoterpene phenols were efficiently encapsulated in NE to enhance their colloidal stability in aqueous solution. The resulting NE showed strong antibacterial activity, reduced inflammation ability, and improved healing of bacteria-infected wounds. TA acted as both an NE stabilizer and a synergistic agent to boost antibacterial effects and accelerate wound repair. This work demonstrates a promising strategy for delivering natural antimicrobial agents of monoterpene phenols to treat infected wounds.