Comparison of Approaches for Authentication of Commercial Terpinen-4-ol-type Tea Tree Oils Using Chiral GC/MS.

A global demand for tea tree oil (TTO) has resulted in increased adulteration in commercial products. In this study, we use a novel enantiomeric gas chromatography mass spectrometry method for chiral analysis of key terpenes ((±)-terpinen-4-ol, (±)-α-terpineol, and (±)-limonene) and quantification of components present at >0.01% to test different methods of identifying adulterated TTO. Data from authentic Australian (n = 88) and oxidized (n = 12) TTO samples of known provenance were consistent with recommended ranges in ISO 4730:2017 and previously published enantiomeric ratios, with p-cymene identified as the major marker of TTO oxidation. The 15 ISO 4730:2017 constituents comprised between 84.5 and 89.8% of the total ion chromatogram (TIC) peak area. An additional 53 peaks were detected in all samples (7.3-11.0% of TIC peak area), while an additional 43 peaks were detected in between 0 and 99% (0.15-2.0% of the TIC peak area). Analysis of nine commercial samples demonstrated that comparison to the ISO 4730:2017 standard does not always identify adulterated TTO samples. While statistical analysis of minor components in TTO did identify two commercial samples that differed from authentic TTO, the (+)-enantiomer percentages for limonene, terpinen-4-ol, and α-terpineol provided clearer evidence that these samples were adulterated. Thus, straightforward identification of unadulterated and unoxidized TTO could be based on analysis of appropriate enantiomeric ratios and quantitation of the p-cymene percentage.

The essential oil of Melaleuca alternifolia incorporated into hydrogel induces antimicrobial and anti-inflammatory effects on infected wounds by Staphylococcus aureus.

Staphylococcus aureus is one of the most common bacterial isolates found in wounds. Thus, innovative dressings, such as hydrogels, are interesting vehicles for incorporating bioactive compounds like those from Melaleuca alternifolia essential oil (MaEO). In this study, we evaluated the antimicrobial and anti-inflammatory potential of MaEO incorporated into an alginate and chitosan hydrogel for treating wounds infected by S. aureus. The hydrogel incorporated with MaEO 1% (HMa 1%) was homogeneous with a bright pale-yellow color and the characteristic smell of Melaleuca. The incorporation of MaEO 1% does not affect the stability of the hydrogel, which was stable up to 90 days of storage. The Scanning electron microscopy analysis revealed that hydrogels showed irregular surfaces and interconnected porous structures with accumulations of oil crystals distributed throughout the formulation. HMa 1% has a high moisture content (95.1%) and can absorb simulated wound fluid. Regarding the antimicrobial effects, HMa 1% reduced the growth of S. aureus ATCC 6538 in both in vitro conditions and in an ex vivo model of wounds using porcine skin. In addition, the dairy topical treatment of murine skin lesions with HMa 1% induced a significant reduction of the wound area, inflammation score, and bacterial load, as well as tissue re-epithelialization and modulation of inflammatory mediators. Therefore, hydrogel incorporated with MaEO 1% has excellent potential to be used in the pharmacotherapy of infected wounds.

Effect of cationic surfactant on the physicochemical and antibacterial properties of colloidal systems (emulsions and microemulsions).

Colloidal systems have been used to encapsulate, protect and release essential oils in mouthwashes. In this study, we investigated the effect of cetylpyridinium chloride (CPC) on the physicochemical properties and antimicrobial activity of oil-in-water colloidal systems containing tea tree oil (TTO) and the nonionic surfactant polysorbate 80. Our main aim was to evaluate whether CPC could improve the antimicrobial activity of TTO, since this activity is impaired when this essential oil is encapsulated with polysorbate 80. These systems were prepared with different amounts of TTO (0-0.5% w/w) and CPC (0-0.5% w/w), at a final concentration of 2% (w/w) polysorbate 80. Dynamic light scattering (DLS) results revealed the formation of oil-swollen micelles and oil droplets as a function of TTO concentration. Increases in CPC concentrations led to a reduction of around 88% in the mean diameter of oil-swollen micelles. Although this variation was of only 20% for the oil droplets, the samples appearance changed from turbid to transparent. The surface charge of colloidal structures was also markedly affected by the CPC as demonstrated by the transition in zeta potential from slightly negative to highly positive values. Electron paramagnetic resonance (EPR) studies showed that this transition is followed by significant increases in the fluidity of surfactant monolayer of both colloidal structures. The antimicrobial activity of colloidal systems was tested against a Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureaus) bacteria. Our results revealed that the inhibition of bacterial growth is observed for the same CPC concentration (0.05% w/w for E. coli and 0.3% w/w for S. aureus) regardless of TTO content. These findings suggest that TTO may not act as an active ingredient in polysorbate 80 containing mouthwashes.

An upcycled fraction of Melaleuca alternifolia essential oil regenerates the skin through the skin melatonin pathway and improves sleep quality.

BACKGROUND: Sleep is one of the most important factors affecting overall health. During the night, the skin repairs damage caused by daily stresses. Melatonin plays a key role in this process. Toxins are removed, and cellular repair and growth hormone production are increased. Inter alia, this also decreases signs of intrinsic aging. AIMS: The current study was intended to demonstrate the impact of a unique fraction of Melaleuca alternifolia (FMA) essential oil, on sleep and skin quality. METHODS: The effect of FMA was investigated in vitro on skin cells, evaluating its antioxidant and anti-inflammatory properties, and in an ex-vivo study on human skin biopsies treated with FMA following stress induction. In addition, two clinical studies were performed on volunteers with life-style-related sleep complaints. In one study, sleep was measured using a noncontact monitoring device (SleepScore Labs, Max). A second study was conducted to assess skin anti-aging effects. RESULTS: In vitro application of FMA reduced IL-8 and reactive oxygen species (ROS) generation in skin cells. This was confirmed ex vivo through a decrease in inflammatory markers and an increase in antioxidant enzymes after stress induction. Interestingly, FMA also upregulated melatonin-associated genes. Real-world sleep tracking revealed that FMA significantly improved sleep quality, relative to unscented control. In vivo applications also showed a reduction in signs of aging. CONCLUSION: These results provide initial data to suggest that this unique FMA delivers skin anti-aging benefits via a two-pronged mode of action, improving sleep quality, and reducing skin inflammatory and oxidative stress.

Nanoemulsions and nanocapsules loaded with Melaleuca alternifolia essential oil for sepsis treatment.

Sepsis represents a complex clinical syndrome that results from a harmful host response to infection. The infections most associated with sepsis are pneumonia, intra-abdominal infection, and urinary tract infection. Tea tree oil (TTO) has shown high antibacterial activity; however, it exhibits low aqueous solubility and high volatility, which have motivated its nanoencapsulation. In this study, the performance of nanoemulsions (NE) and nanocapsules (NC) loaded with TTO was compared. These systems were prepared by spontaneous emulsification and nanoprecipitation methods, respectively. Poly-ε-caprolactone or Eudragit® RS100 were tested as polymers for NCs whereas Tween® 80 or Pluronic® F68 as surfactants in NE preparation. Pluronic® F68 and Eudragit® RS100 resulted in more homogeneous and stable nanoparticles. In accelerated stability studies at 4 and 25 °C, both colloidal suspensions (NC and NE) were kinetically stable. NCs showed to be more stable to photodegradation and less cytotoxic than NEs. After sepsis induction by the cecal ligation and puncture (CLP) model, both NE and NC reduced neutrophil infiltration into peritoneal lavage (PL) and kidneys. Moreover, the systems increased group thiols in the kidney and lung tissue and reduced bacterial growth in PL. Taken together, both systems showed to be effective against injury induced by sepsis; however, NCs should be prioritized due to advantages in terms of cytotoxicity and physicochemical stability.

Fungicide potential of citronella and tea tree essential oils against tomato cultivation's phytopathogenic fungus Fusarium oxysporum f. sp. lycopersici and analysis of their chemical composition by GC/MS.

Tomato is one of the most produced and consumed fruits in the world. However, it is a crop that faces several phytosanitary problems, such as fusarium wilt, caused by Fusarium oxysporum. Thus, this study aimed to evaluate citronella and melaleuca essential oils in vitro potential in the fungus F. oxysporum management. The chemical identification of the components in the essential oils was performed by gas chromatography with flame ionization and mass spectrometer detectors. The IC50 and IC90 were determined by linear regression and the percentage of inhibition of the fungus by analysis of variance. The major compounds in citronella essential oil were citronellal, Geraniol, and citronellol; in melaleuca (tea tree) oil were terpinen-4-ol and α-terpinene. Both oils promoted more significant inhibition at concentrations of 1.5 and 2.5 µL/mL, besides not presenting significant differences with commercial fungicides, confirming the high potential for using this control method in agriculture.

Effect of Tea Tree Oil Addition to Denture Liners Against Candida albicans and Bond Strength to Acrylic Denture Bases.

PURPOSE: To evaluate the effect of adding tea tree oil to denture liners on Candida albicans and bond strength to the acrylic denture base. MATERIALS AND METHODS: Disc-shaped specimens were fabricated from silicone-based resilient liner (Tokuyama, Molloplast), acrylic-based hard liner (GC Reline), and acrylic-based soft liner (Visco-gel). Tea tree oil (TTO) was incorporated into the liners at varying concentrations (0% [control], 2%, 5%, 8%). C albicans were counted by viable colony count, and optical density (OD) was measured with a spectrophotometer. The tensile strength to heat polymerized acrylic denture base was measured in a universal testing machine. The compliance of the data to the distribution of normality was evaluated using the Shapiro Wilk test. Two-way ANOVA, Bonferroni correction, and paired sample t test were performed (α = .05). RESULTS: The addition of TTO into liners provided a significant decrease in the OD values (P < .001). The control groups of the liners presented the highest colony counts, whereas increasing TTO decreased the results (P < .01). According to tensile bond strength test, 8% TTO addition resulted in a significant decrease for Tokuyama (P < .01) and Molloplast liners (P < .05), while 2% TTO resulted in significance for GC Reline (P < .001). CONCLUSIONS: Denture liners containing increasing percentages of TTO presented lower amounts of C albicans colonies and decreased bond strength to the denture bases. When using TTO for its antifungal properties, the amount added should be carefully selected because the tensile bond strength may be affected.

Volatile organic compounds (VOCs) of essential oils for the control of Fusarium oxysporum in cherry tomato seeds.

Fusarium oxysporum is the causal agent of Fusarium wilt in tomato plants. The most common form of control of this disease is through seed chemical treatment. However, the present work presents an alternative method, through the fumigation technique with essential oils. The pathogen F. oxysporum was inoculated on organic cherry tomato seeds through contact with sporulated Petri® plates. Thereafter, seeds were placed in stainless steel crucibles containing a 1.0 x 1.0 cm filter paper adhered to the lid and kept for 24 hours. This paper received 20 µL of each essential oil: tea tree, chia, citronella, lavender, anise basil, clove basil, and deionized water as control. This process was called "seed fumigation by essential oil". After this process, a germination test was carried out in germ boxes with Germitest® paper to verify the variables Germination Speed Index (GSI), Germination (G%), and Mean time to germination (MGT). Mycelial growth was verified in Petri® plates containing PDA medium. The plates containing mycelial growth were observed through scanning electron microscopy to verify possible morphological damage in the hyphae of the pathogen. Tea tree essential oil was the one that allowed the greatest suppression of the phytopathogen. Therefore, new tests were carried out with this specific oil. In germ boxes, tests of germination (G%), Abnormal seedlings count (ASC), and percentage of seedlings with mycelial growth were carried out. In addition, plant elicitation tests were performed in tomato seedlings through the analysis of chitinase, glucanase, and total proteins. All tests were carried out in completely randomized designs with four replications. All data were submitted to the Lilliefors normality test, followed by the analysis of variance, and Tukey's HSD (5% significance) for mean comparison. It was found that tea tree essential oil inhibited the mycelial growth of F. oxysporum without affecting the germination of cherry tomato seeds. Subsequent tests with this oil also demonstrated that there is a reduction in mycelia present in the seeds and a reduction in abnormal seedlings compared to the control. There was no significant difference between the variables tested for plant elicitation.

Comparison of chitosan and SLN nano-delivery systems for antibacterial effect of tea tree (Melaleuca alternifolia) oil against P. aeruginosa and S. aureus.

Treatment of wounds is challenging due to bacterial infections, including Staphylococcus aureus and Pseudomonas aeruginosa. Using the merits of alternative antimicrobials like tea tree oil (TTO) and nanotechnology, they can be helpful in combatting bacterial infections. Solid lipid nanoparticle (SLN) and chitosan (CS) nanoparticles show great potential as carriers for enhancing the stability and therapeutic benefits of oils. The aim of this study is to compare the influence of nanocarriers in enhancing the antibacterial effects of TTO. The study evaluates the physicochemical and antibacterial properties of TTO-SLN and TTO-CS against P. aeruginosa and S. aureus. The TTO-SLN nanoparticles showed a clear round shape with the average diameter size of 477 nm, while the TTO-CS nanoparticles illustrated very homogeneous morphology with 144 nm size. The encapsulation efficiency for TTO-CS and TTO-SLN was ∼88.3% and 73.5%, respectively. Minimum inhibitory concentration against S. aureus and P. aeruginosa for TTO-CS, TTO-SLN, and pure TTO were 35 and 45 µg ml-1, 130 and 170 µg ml-1, and 380 and 410 µg ml-1, respectively. Since TTO-CS revealed an impressively higher antimicrobial effects in comparison with TTO-SLN and TTO alone, it can be considered as a nanocarrier that produces the same antimicrobial effects with lower required amounts of the active substance.

Antimicrobial and antibiofilm effects of shikonin with tea tree oil nanoemulsion against Candida albicans and Staphylococcus aureus.

The current work aims to develop a shikonin and tea tree oil loaded nanoemulsion system stabilized by a mixture of GRAS grade surfactants (Tween 20 and monoolein) and a cosurfactant (Transcutol P). This system was designed to address the poor aqueous solubility and photostability issues of shikonin. The authenticity of shikonin employed in this study was confirmed using nuclear magnetic resonance (NMR) spectroscopy. The optimized nanoemulsion exhibited highly favorable characteristics in terms of zeta potential (-23.8 mV), polydispersity index (0.216) and particle size (22.97 nm). These findings were corroborated by transmission electron microscopy (TEM) micrographs which confirmed the spherical and uniform nature of the nanoemulsion globules. Moreover, attenuated total reflectance (ATR) and X-ray diffraction analysis (XRD) analysis affirmed improved chemical stability and amorphization, respectively. Photodegradation studies were performed by exposing pure shikonin and the developed nanoemulsion to ultraviolet light for 1 h using a UV lamp, followed by high performance liquid chromatography (HPLC) analysis. The results confirmed that the developed nanoemulsion system imparts photoprotection to pure shikonin in the encapsulated system. Furthermore, the research investigated the effect of the nanoemulsion on biofilms formed by Candida albicans and methicillin resistant Staphylococcus aureus (MRSA). Scanning electron microscopy, florescence microscopy and phase contrast microscopy unveiled a remarkable reduction in biofilm area, accompanied by disruptions in the cell wall and abnormalities on the cell surface of the tested microorganisms. In conclusion, the nanoencapsulation of shikonin with tea tree oil as the lipid phase showcased significantly enhanced antimicrobial and antibiofilm potential compared to pure shikonin against resistant strains of Candida albicans and Staphylococcus aureus.

Effects of adding tea tree oil on growth performance, immune function, and intestinal function of broilers.

The aim of this study was to investigate the effects of adding tea tree oil (TTO) in the basal diet on growth performance, immune function, and intestinal function in broilers. This study utilized 1,650 one-day-old broilers with good health and similar body weight. Subjects were randomized into 5 groups with 6 replicates each: the control group (CON, basal diet), positive control group (PCG, basal diet + 100 mg/kg oregano oil in diet), low-dose TTO group (TTO-L, 50 mg/kg TTO added in the basal diet), medium-dose TTO group (TTO-M, 100 mg/kg TTO added in the basal diet), and high-dose TTO group (TTO-H, 200 mg/kg TTO added in the basal diet). The whole test period lasted 28 d. The results showed that the broilers fed with TTO supplemented diet had significantly higher body weight and average daily gain (ADG) (P = 0.013), and had a lower feed conversion ratio (F/G) (P = 0.010) throughout the trial period. The index of thymus in TTO-M increased significantly compared to CON (P = 0.015) on d 28. On d 14 and 28, C3, IFN-γ, TNF-α, and IL-2 levels in TTO-L serum were significantly increased (P < 0.001); the 3 test groups supplemented with TTO had significantly higher titers of avian influenza H9 subtype in their serum (P < 0.05). Tea tree oil supplement in the diet also had a positive and significant effect on the intestinal morphology of broilers throughout the experiment (P < 0.05). These results indicate that TTO has the ability to promote broiler growth, regulate immunity, and improve intestinal morphology. The proposed dosage of adding 50 mg/kg in broiler basal diets provides a theoretical basis for its subsequent use in livestock feeds.

Tea Tree Essential Oil Kills Escherichia coli and Staphylococcus epidermidis Persisters.

Persister cells are a small subpopulation of non-growing bacteria within a population that can survive long exposures to antibiotic treatment. Following antibiotic removal, persister cells can regrow and populate, playing a key role in the chronic reoccurrence of bacterial infections. The development of new molecules and methods to kill bacterial persisters is critical. Essential oils and other natural products have long been studied for their antimicrobial effects. Here, we studied the effectiveness of tea tree essential oil (TTO), a common component in many commercial care products, against Escherichia coli and Staphylococcus epidermidis persister cells. Using biphasic kill curve assays, we found that concentrations of 0.5% and 1.0% TTO for E. coli and S. epidermidis, respectively, completely eradicated persister cells over a period of 24 h, with the component terpinen-4-ol responsible for most of the killing. Using a colorimetric assay, it was determined that the TTO exhibited its anti-persister effects through a membrane disruption mechanism.

Nanoemulsions (O/W) prepared from essential oil extracted from Melaleuca alternifolia: synthesis, characterization, stability and evaluation of anticancerous, anti-oxidant, anti-inflammatory and antidiabetic activities.

Essential oil from Melaleuca alternifolia (also known as Tea tree essential oil, TTO) is used as traditional medicine and used as therapeutic in medicine, food and cosmetic sectors. However, this oil is highly unstable, volatile and prone to oxidation which limits its practical use. The objective of this study was synthesis of tea tree oil based O/W (oil/water) nanoemulsions (tea tree essential oil nanoemulsions, TNE) and evaluation of its biological potential. Physiological characterization was carried out using UV, fluorescent, and FT-IR techniques. Various biological activities such as anticancerous, antidiabetic and anti-inflammatory were also estimated. Pharmacokinetics study on TNE was carried out. Encapsulation efficiency of nanoemulsions was found to be 83%. Nanoemulsions were spherical in shape with globule size 308 nm, zeta potential -9.42 and polydispersity index was 0.31. Nanoemulsions were stable even after 50 days of storage at different temperatures. Anti-oxidant potential of TNE was conducted by various assays and IC50 were: Nitric oxide radical scavenging activity:225.1, DPPH radical scavenging activity:30.66, Iron chelating assay:38.73, and Iron reducing assay:39.36. Notable anticancer activity was observed with the percent cell viability of HeLa cells after treatment with 1, 2 and 5 µl of TNE was 82%, 41% and 24%, respectively. Antidiabetic study revealed that TNE inhibited -amylase in a dose-dependent manner, with 88% inhibition at its higher volume of 250 µl. Drug kinetic study revealed that nanoemulsions exhibited first-order model. Based on this, the possible role of M. alternifolia oil-based nanoemulsions in cosmetic, food, and pharma sectors has been discussed.

Fabrication and Biological Activities of All-in-One Composite Nanoemulsion Based on Blumea balsamifera Oil-Tea Tree Oil.

Nanoemulsion is a new multi-component drug delivery system; the selection of different oil phases can give it special physiological activity, and play the role of "medicine and pharmaceutical excipients all-in-one". In this paper, we used glycyrrhizic acid as the natural surfactant, and Blumea balsamifera oil (BB) and tea tree oil (TTO) as the mixed oil phase, to obtain a new green functional composite nanoemulsion. Using the average particle size and polydispersion index (PDI) as the evaluation criteria, the effects of the oil ratio, oil content, glycyrrhizic acid concentration, and ultrasonic time on the nanoemulsion were systematically investigated. The stability and physicochemical properties and biological activities of BB-TTO NEs prepared via the optimum formulation were characterized. The optimal prescription was BB: TTO = 1:1, 5% oil phase, 0.7% glycyrrhizic acid, and 5 min ultrasonication time. The mean particle size, PDI, and zeta potential were 160.01 nm, 0.125, and -50.94 mV, respectively. The nanoemulsion showed non-significant changes in stability after centrifugation, dilution, and 120 days storage. These nanoemulsions were found to exhibit potential antibacterial and anti-inflammatory activities. The minimal inhibitory concentration (MIC) of BB-TTO NEs against Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa is 2975 µg/mL, 2975 µg/mL, and 5950 µg/mL, respectively. A lower level of inflammatory cell infiltration and proportion of fibrosis were found in the synovial tissue of AIA rats treated with BB-TTO NEs. These findings demonstrate that the BB-TTO NEs produced in this study have significant potential for usage in antibacterial and anti-inflammatory areas.

Tea tree oil, a vibrant source of neuroprotection via neuroinflammation inhibition: a critical insight into repurposing Melaleuca alternifolia by unfolding its characteristics. / èŒ¶æ ‘æ²¹æŠ‘åˆ¶ç¥žç»ç‚Žç—‡å¹¶æä¾›ç¥žç»ä¿æŠ¤­­æŽ¢ç´¢äº’花千层的潜力.

Over the past few decades, complementary and alternative treatments have become increasingly popular worldwide. The purported therapeutic characteristics of natural products have come under increased scrutiny both in vitro and in vivo as part of efforts to legitimize their usage. One such product is tea tree oil (TTO), a volatile essential oil primarily obtained from the native Australian plant, Melaleuca alternifolia, which has diverse traditional and industrial applications such as topical preparations for the treatment of skin infections. Its anti-inflammatory-linked immunomodulatory actions have also been reported. This systematic review focuses on the anti-inflammatory effects of TTO and its main components that have shown strong immunomodulatory potential. An extensive literature search was performed electronically for data curation on worldwide accepted scientific databases, such as Web of Science, Google Scholar, PubMed, ScienceDirect, Scopus, and esteemed publishers such as Elsevier, Springer, Frontiers, and Taylor & Francis. Considering that the majority of pharmacological studies were conducted on crude oils only, the extracted data were critically analyzed to gain further insight into the prospects of TTO being used as a neuroprotective agent by drug formulation or dietary supplement. In addition, the active constituents contributing to the activity of TTO have not been well justified, and the core mechanisms need to be unveiled especially for anti-inflammatory and immunomodulatory effects leading to neuroprotection. Therefore, this review attempts to correlate the anti-inflammatory and immunomodulatory activity of TTO with its neuroprotective mechanisms.

Allergic contact dermatitis from essential oil in consumer products: Mode of uses and value of patch tests with an essential oil series. Results of a French study of the DAG (Dermato-Allergology group of the French Society of Dermatology).

OBJECTIVE: To analyse the clinical characteristics and sensitivity of an essential oil patch test series (EOS) in patients sensitized to their own essential oils (EOs). METHOD: We analysed the clinical data and patch test results obtained with the European baseline series (BSE) and an EOS, as well as the mode of use of EOs, through a questionnaire included in the patient file. RESULTS: The study included 42 patients (79% women, average age 50 years) with allergic contact dermatitis (ACD), 8 patients required hospitalization. All patients were sensitized to the EO they used, primarily lavender (Lavandula augustifolia, 8000-28-0), tea tree (Melaleuca alternifolia leaf oil, 68647-73-4), ravintsara (Cinnamomum camphora oil, 92201-50-8), and 2 cases were attributed to helichrysum (helichrysum italicum flower absolute, 90045-56-0). 71% had positive patch tests to fragrance mix I or II, 9 only to the EOS and 4 only with their personal EO. Interestingly, 40% of patients did not spontaneously mention the use of EOs, and only 33% received advice on their use at the time of purchase. CONCLUSION: Patch tests with the BSE, limonene and linalool HP, and oxidized tea tree oil is sufficient to detect most EO-sensitized patients. The most important is to test the patient's own used EOs.

The synergy of tea tree oil nano-emulsion and antibiotics against multidrug-resistant bacteria.

AIMS: We determined the synergistic effects of tea tree essential oil nano-emulsion (nanoTTO) and antibiotics against multidrug-resistant (MDR) bacteria in vitro and in vivo. Then, the underlying mechanism of action of nanoTTO was investigated. METHODS AND RESULTS: Minimum inhibitory concentrations and fractional inhibitory concentration index (FICI) were determined. The transepithelial electrical resistance (TEER) and the expression of tight junction (TJ) protein of IPEC-J2 cells were measured to determine the in vitro efficacy of nanoTTO in combination with antibiotics. A mouse intestinal infection model evaluated the in vivo synergistic efficacy. Proteome, adhesion assays, quantitative real-time PCR, and scanning electron microscopy were used to explore the underlying mechanisms. Results showed that nanoTTO was synergistic (FICI ≤ 0.5) or partial synergistic (0.5 < FICI < 1) with antibiotics against MDR Gram-positive and Gram-negative bacteria strains. Moreover, combinations increased the TEER values and the TJ protein expression of IPEC-J2 cells infected with MDR Escherichia coli. The in vivo study showed that the combination of nanoTTO and amoxicillin improved the relative weight gain and maintained the structural integrity of intestinal barriers. Proteome showed that type 1 fimbriae d-mannose specific adhesin of E. coli was downregulated by nanoTTO. Then, nanoTTO reduced bacterial adhesion and invasion and inhibited the mRNA expression of fimC, fimG, and fliC, and disrupted bacterial membranes.

Electrospun chitosan-based nanofibers loading tea tree oil for fresh salmon fillet shelf-life extension.

Bioactive packaging can improve the shelf-life of food products and enhance consumer health. It can also alleviate environmental stress on the planet by reducing food waste. Here, the electrospinning of tea tree oil-loaded 2-hydroxypropyltrimethyl ammonium chloride chitosan nanofibers was investigated. The fabricated nanofiber films were characterized by scanning electron microscopy, thermal gravimetric analysis, Fourier transform infrared spectroscopy, and contact angle meter analysis. The prepared nanofibers have a well-defined diameter of about 200 nm and a smooth shape. They have good antibacterial properties against Staphylococcus aureus and Escherichia coli in vitro. Tea tree oil-loaded chitosan-based nanofibers were found to be effective in delaying spoilage and extending the shelf life of salmon by sensory evaluation, texture analysis, color, total viable counts, thiobarbituric acid, and total volatile basic nitrogen during storage in the freshness experiments, thus indicating their health benefits in bioactive packaging.

The Safety Assessment of Cosmetic Perfumes by Using In Chemico and In Vitro Methods in Combination with GC-MS/MS Analysis.

Animal testing has been prohibited for the safety assessment of cosmetic ingredients or finished products. Thus, alternative non-animal methods, followed by confirmatory clinical studies on human volunteers, should be used as the sole legally acceptable approach within the EU. The safety assessment of cosmetic products requires the involvement of multiple scientific disciplines, including analytical chemistry and biomedicine, as well as in chemico, in vitro and in silico toxicology. Recent data suggest that fragrance components may exert multiple adverse biological effects, e.g. cytotoxicity, skin sensitisation, (photo)genotoxicity, mutagenicity, reprotoxicity and endocrine disruption. Therefore, a pilot study was conducted with selected samples of fragrance-based products, such as deodorant, eau de toilette and eau de parfum, with the aim of integrating results from a number of alternative non-animal methods suitable for the detection of the following toxicological endpoints: cytotoxicity (with 3T3 Balb/c fibroblasts); skin sensitisation potential (in chemico method, DPRA); skin sensitisation potential (LuSens in vitro method, based on human keratinocytes); genotoxicity potential (in vitro Comet assay with 3T3 Balb/c cells); and endocrine disruption (in vitro YES/YAS assay). The presence of twenty-four specific known allergens in the products was determined by using GC-MS/MS. The strategies for estimation of the NOAEL of a mixture of allergens, which were proposed by the Scientific Committee on Consumer Products in their 'Opinion on Tea tree oil' document and by the Norwegian Food Safety Authority in their 'Risk Profile of Tea tree oil' report, were used as models for the NOAEL estimation of the mixtures of allergens that were identified in the individual samples tested in this study.

Eco-friendly and effective antimicrobial Melaleuca alternifolia essential oil Pickering emulsions stabilized with cellulose nanofibrils against bacteria and SARS-CoV-2.

Melaleuca alternifolia essential oil (MaEO) is a green antimicrobial agent suitable for confection eco-friendly disinfectants to substitute conventional chemical disinfectants commonly formulated with toxic substances that cause dangerous environmental impacts. In this contribution, MaEO-in-water Pickering emulsions were successfully stabilized with cellulose nanofibrils (CNFs) by a simple mixing procedure. MaEO and the emulsions presented antimicrobial activities against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli). Moreover, MaEO deactivated the SARS-CoV-2 virions immediately. FT-Raman and FTIR spectroscopies indicate that the CNF stabilizes the MaEO droplets in water by the dipole-induced-dipole interactions and hydrogen bonds. The factorial design of experiments (DoE) indicates that CNF content and mixing time have significant effects on preventing the MaEO droplets' coalescence during 30-day shelf life. The bacteria inhibition zone assays show that the most stable emulsions showed antimicrobial activity comparable to commercial disinfectant agents such as hypochlorite. The MaEO/water stabilized-CNF emulsion is a promissory natural disinfectant with antibacterial activity against these bacteria strains, including the capability to damage the spike proteins at the SARS-CoV-2 particle surface after 15 min of direct contact when the MaEO concentration is 30 % v/v.