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This work aimed to explore an alternative to the use of antibiotics for prevention and treatment of wounds infection caused by two common bacterial pathogens Staphylococcus aureus and Pseudomonas aeruginosa. For this purpose, three different essential oil components (EOCs), namely carvacrol, citronellol and cinnamic acid, were loaded into electrospun fibers of poly-ε-caprolactone (PCL) aided by alpha-cyclodextrin (αCD) and hydroxypropyl-ß-cyclodextrin (HPßCD). Electrospun-fibers prepared with each EOC and their mixtures were screened for antimicrobial capability and characterized regarding morphological, mechanical, thermal, surface polarity, antibiofilm and antioxidant properties. αCD formed poly(pseudo)rotaxanes with PCL and weakly interacted with EOCs, while HPßCD facilitated EOC encapsulation and formation of homogeneous fibers (500-1000 nm diameter) without beads. PCL/HPßCD fibers with high concentration of EOCs (mainly carvacrol and cinnamic acid) showed strong antibiofilm (>3 log CFU reduction) and antioxidant activity (10-50% DPPH scavenging effects). Different performances were recorded for the EOCs and their mixtures; cinnamic acid migrated to fiber surface and was released faster. Fibers biocompatibility was verified using hemolysis tests and in ovo tissue integration and angiogenesis assays. Overall, HPßCD facilitates complete release of EOCs from the fibers to the aqueous medium, being an environment-friendly and cost-effective strategy for the treatment of infected wounds.
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Monoterpenos Acíclicos , Antioxidantes , Cinamatos , Cimenos , Monoterpenos , Cicatrización de Heridas , Cicatrización de Heridas/efectos de los fármacos , Cimenos/farmacología , Cimenos/química , Cinamatos/química , Cinamatos/farmacología , Monoterpenos Acíclicos/farmacología , Monoterpenos Acíclicos/química , Antioxidantes/farmacología , Antioxidantes/química , Monoterpenos/farmacología , Monoterpenos/química , Antiinfecciosos/farmacología , Antiinfecciosos/química , Staphylococcus aureus/efectos de los fármacos , Ciclodextrinas/química , Ciclodextrinas/farmacología , Biopelículas/efectos de los fármacos , Nanofibras/química , Pruebas de Sensibilidad Microbiana , Antibacterianos/farmacología , Antibacterianos/química , Poliésteres/química , Aceites Volátiles/farmacología , Aceites Volátiles/química , Pseudomonas aeruginosa/efectos de los fármacosRESUMEN
Dermatophytosis is a prevalent fungal infection and public health burden, majorly caused by the attack of zoophilic fungi genera of Trichophyton and Microsporum. Among them, T. mentagrophytes and M. canis are the dominating pathogens that cause dermatophytosis in humans. Though anti-fungal treatments are available, the widespread drug resistance and minimal efficacy of conventional therapies cause recurring infections. In addition, prolonged anti-fungal medications induce several systemic side effects, including hepatotoxicity and leucopenia. The anti-dermatophytic formulation of biocompatible essential oil components (EOCs) is attractive due to their highly potent anti-dermatophytic action. Herein, two EOCs, Eugenol (EU) and Isoeugenol (IU), incorporated emulsion hydrogel (EOCs-EHG) synthesized from hydroxypropylmethyl cellulose and poly(ethylene glycol) methyl ether methacrylate. The cytocompatibility of the hydrogels is confirmed by treating them with fibroblast and keratinocyte cell lines. The EOCs-EHG demonstrated pH and temperature-responsive sustained release of entrapped EOCs and inhibited fungal spore germination. T. mentagrophytes and M. canis biofilms are eradicated at a minimal inhibitory concentration of 2 µg mL-1 each of EU and IU. The in vivo anti-dermatophytic activity of EOCs-EHG is confirmed in dermatophyte-infected Wistar albino rat models. The topical application of EOCs-EHG demonstrated complete infection eradication and facilitated skin regeneration, emphasizing the therapeutic potential of EOCs-EHG against dermatophytosis.
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Using existing adrentimicrobials with essential oil components to prevent antimicrobial resistance is an alternative strategy. This study aimed to evaluate the resistance status, synergistic combinations, and in vitro biofilm formation activities of clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA), Stenotrophomonas maltophilia and Candida albicans against antimicrobial agents and cinnamaldehyde, carvacrol, eugenol, limonene and eucalyptol. Antimicrobial activities were evaluated by microdilution, cytotoxicity by XTT, synergy by checkerboard and time-kill, and biofilm inhibition by microplate methods. Cinnamaldehyde and carvacrol showed strong antimicrobial activity. Synergistic effects were observed when using all essential oils with antimicrobials. Only two C. albicans isolates showed antagonism with cinnamaldehyde and fluconazole. The constituents showed cytotoxic effects in the L929 cell line (except limonene). A time-kill analysis revealed a bacteriostatic effect on S. maltophilia and MRSA isolates and a fungicidal effect on C. albicans isolates. These results are important for further research to improve antimicrobial efficacy or to develop new agents.
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Antiinfecciosos , Biopelículas , Candida albicans , Sinergismo Farmacológico , Staphylococcus aureus Resistente a Meticilina , Pruebas de Sensibilidad Microbiana , Aceites Volátiles , Stenotrophomonas maltophilia , Biopelículas/efectos de los fármacos , Aceites Volátiles/farmacología , Aceites Volátiles/química , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Staphylococcus aureus Resistente a Meticilina/fisiología , Candida albicans/efectos de los fármacos , Candida albicans/fisiología , Stenotrophomonas maltophilia/efectos de los fármacos , Stenotrophomonas maltophilia/fisiología , Antiinfecciosos/farmacología , Limoneno/farmacología , Acroleína/análogos & derivados , Acroleína/farmacología , Cimenos/farmacología , Línea Celular , Monoterpenos/farmacología , Antibacterianos/farmacología , Terpenos/farmacología , Eucaliptol/farmacología , Eugenol/farmacología , Ciclohexenos/farmacología , RatonesRESUMEN
Extracellular phospholipase (EPL) plays an important role in the pathogenesis of the yeast Malassezia pachydermatis. Currently, the attention of researchers is focused on studying the virulence factors involved in this process and searching solutions to reduce their activity. One of the options is the use of natural remedies as anti-virulence agents. This study is aimed at investigating the production of extracellular phospholipase in M. pachydermatis strains (18 samples) and followed by the time-dependent inhibitory effect of selected azole antifungals (itraconazole, posaconazole and voriconazole) and plant essential oil components (terpinen-4-ol, thymol, carvacrol, eugenol and geraniol), evaluated by Egg Yolk Agar plate method. Almost all strains (17 isolates, (94.4%) were found to be intense EPL producers. A significant, time-dependent inhibition of EPL was noted after 1-, 3- and 6-h exposure of Malassezia cells to itraconazole (26.4%, 47.2% and 50.9%, respectively) compared to exposure to posaconazole (26.4%, 28.3% and 28.3%, respectively) and voriconazole (18.8%, 20.8% and 35.8%, respectively). After one-hour exposure to plant essential oil components, the best inhibitory effect was recorded for eugenol (62.3%), followed by terpinen-4-ol and thymol (56.6%), geraniol (41.5%) and carvacrol (26.4%). A 3-h exposure revealed that thymol retained the best inhibitory effect (88.7%) on EPL production, followed by carvacrol (73.6%), eugenol (56.6%), terpinen-4-ol (52.8%) and geraniol (49.1%). After 6-h exposure, no growth of M. pachydermatis strains exposed to carvacrol was observed, and the inhibitory efficiency for the other tested essential oil (EO) components achieved 88.7%. The obtained results indicate the promising efficacy of plant essential oils components in the inhibition of virulence factors such as EPL production.
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AIM: In this study, it was aimed to examine the antibacterial activity of the essential oil components (EOCs), carvacrol (CAR), cinnamaldehyde (CIN), thymol (TH), alpha pinene (α-PN), eucalyptol (EU), limonene (LIM), and the antibiotics, linezolid (LZD), vancomycin (VAN), gentamicin (GEN), ciprofloxacin (CIP), clindamycin (CLN), and penicillin (PEN) against 50 multidrug resistant Corynebacterium striatum strains, and the synergistic interactions of CAR and CIN with the antibiotics against 10 randomly selected Coryne. striatum strains to explore synergistic interactions to determine if their combined use could enhance antibiotic activity and potentially reduce resistance. METHODS AND RESULTS: The activity of the EOCs and the antibiotics against Coryne. striatum strains isolated from clinical specimens, was examined by broth microdilution method. The synergistic interactions of the EOCs with the antibiotics against 10 randomly selected Coryne. striatum strains were determined by checkerboard method. EOCs, CIN, and CAR and antibiotics, LZD, VAN, GEN, CIP, and CLN were detected to have antibacterial activity against Coryne. striatum strains alone and either synergistic interactions were observed in combinations of the antibiotics with EOCs. CONCLUSIONS: All Coryne. striatum strains were determined to be susceptible to VAN and LZD and resistant to GEN, PEN, CIP, and CLN. Synergistic interactions were observed in all combinations of antibiotics tested with CAR and CIN.
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Acroleína , Acroleína/análogos & derivados , Antibacterianos , Corynebacterium , Farmacorresistencia Bacteriana Múltiple , Sinergismo Farmacológico , Pruebas de Sensibilidad Microbiana , Monoterpenos , Aceites Volátiles , Antibacterianos/farmacología , Corynebacterium/efectos de los fármacos , Aceites Volátiles/farmacología , Farmacorresistencia Bacteriana Múltiple/efectos de los fármacos , Acroleína/farmacología , Monoterpenos/farmacología , Cimenos/farmacología , Ciprofloxacina/farmacología , Gentamicinas/farmacología , Vancomicina/farmacología , Linezolid/farmacología , Limoneno/farmacología , Eucaliptol/farmacología , Timol/farmacología , Clindamicina/farmacología , Humanos , Penicilinas/farmacología , Terpenos/farmacología , Ciclohexenos/farmacología , Infecciones por Corynebacterium/microbiologíaRESUMEN
Lowering dietary protein content is a promising strategy to reduce N excretions in cattle but it requires improved N utilization by the animal. Feed enzymes (e.g., exogenous α-amylase) and plant extracts (e.g., essential oils [EO]) are 2 additives that may enhance rumen function and possibly also microbial protein yield. This may increase fat- and protein-corrected milk yield (MY) and milk nitrogen efficiency and thus lower N losses from dairy cows. Both types of additives were studied in an experiment including 39 Holstein cows that had (average ± SD) 40.7 ± 7.95 kg/d MY, 89 ± 43 DIM, 2.7 ± 1.5 lactations, and 677 ± 68.6 kg of BW, consisting of a covariate (4 wk) and treatment period (5 wk). During the whole experiment cows were fed a typical Benelux diet (CTRL), supplemented with concentrates to meet individual requirements for energy and MP, which were fulfilled for 100% and 101%, respectively. The total diet was low in CP (15.5%) and relatively high in starch (22.6% and 6.6% rumen bypass starch). Cows were balanced for parity, DIM, MY, and roughage intake and randomly assigned to one of 3 groups, which received the following treatments in the treatment period: (1) CTRL (n = 13); (2) CTRL + 14 g/cow per day Ronozyme RumiStar α-amylase enzyme (AMEZ, n = 13; DSM); and (3) CTRL + 2.5 g/cow per day Crina Protect, a blend of EO components (ESOL, n = 13; DSM). Animal performance, ruminal pH, and enteric gas emissions were monitored throughout the experiment. During the last week of the covariate and treatment periods, nitrogen balances were conducted, total-tract nutrient digestibility was determined, and urinary allantoin and uric acid were determined as indicators for microbial N production. The statistical model applied to these variables contained group and DIM during treatment period as fixed effects and the values from the covariate period as covariate. Post hoc Dunnet-corrected comparisons between each treatment group and the control group were explored. The α-amylase enzyme tended to increase apparent total-tract starch digestibility and increased milk lactose concentration. The EO blend tended to increase MY and increased milk N output, milk nitrogen efficiency, and feed efficiency. Therefore, when feeding reduced dietary protein levels, EO have potential to improve the N-use efficiency in cattle, whereas the α-amylase enzyme might increase starch digestibility and milk lactose. However, additional research is necessary to substantiate our findings.
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Alimentación Animal , Suplementos Dietéticos , Nitrógeno , Aceites Volátiles , alfa-Amilasas , Animales , Bovinos , Femenino , alfa-Amilasas/metabolismo , Alimentación Animal/análisis , Fenómenos Fisiológicos Nutricionales de los Animales , Dieta/veterinaria , Digestión/efectos de los fármacos , Lactancia , Leche/química , Nitrógeno/metabolismo , Nutrientes/metabolismo , Rumen/metabolismoRESUMEN
BACKGROUND: Treatment of Candida albicans associated infections is often ineffective in the light of resistance, with an urgent need to discover novel antimicrobials. Fungicides require high specificity and can contribute to antifungal resistance, so inhibition of fungal virulence factors is a good strategy for developing new antifungals. OBJECTIVES: Examine the impact of four plant-derived essential oil components (1,8-cineole, α-pinene, eugenol, and citral) on C. albicans microtubules, kinesin motor protein Kar3 and morphology. METHODS: Microdilution assays were used to determine minimal inhibitory concentrations, microbiological assays assessed germ tube, hyphal and biofilm formation, confocal microscopy probed morphological changes and localization of tubulin and Kar3p, and computational modelling was used to examine the theoretical binding of essential oil components to tubulin and Kar3p. RESULTS: We show for the first time that essential oil components delocalize the Kar3p, ablate microtubules, and induce psuedohyphal formation with reduced biofilm formation. Single and double deletion mutants of kar3 were resistant to 1,8-cineole, sensitive to α-pinene and eugenol, but unimpacted by citral. Strains with homozygous and heterozygous Kar3p disruption had a gene-dosage effect for all essential oil components, resulting in enhanced resistance or susceptibility patterns that were identical to that of cik1 mutants. The link between microtubule (αß-tubulin) and Kar3p defects was further supported by computational modeling, showing preferential binding to αß-tubulin and Kar3p adjacent to their Mg2+-binding sites. CONCLUSION: This study highlights how essential oil components interfere with the localization of the kinesin motor protein complex Kar3/Cik1 and disrupt microtubules, leading to their destabilization which results in hyphal and biofilm defects.
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Aceites Volátiles , Proteínas de Saccharomyces cerevisiae , Candida albicans/metabolismo , Cinesinas/metabolismo , Proteínas Asociadas a Microtúbulos/metabolismo , Tubulina (Proteína)/genética , Tubulina (Proteína)/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/metabolismo , Aceites Volátiles/farmacología , Eugenol/metabolismo , Eucaliptol/metabolismo , Microtúbulos/metabolismo , Antifúngicos/farmacología , Antifúngicos/metabolismo , Proteínas de Microtúbulos/metabolismoRESUMEN
In this study, the effects of altitude and soil properties on the essential oil amounts and essential oil components of the Turkish sage (Salvia fruticosaMill.) plant were investigated. A location in the south of Turkey in the Kas district of Antalya with altitude ranges of 0-200 m, 300-500 m, and 600-800 m was defined as the study area. During the full flowering period of S. fruticosa, plants and soil samples were collected. The contents of the essential oils of the plants were analyzed using GC-MS. According to the results obtained, the essential oil content of plants varied between 1.91% and 5.10%. The main component of the essential oils of the plants collected from all study areas was 1,8-cineol, at 44.93%. It was concluded that variations in the proportions of the components limonene and linalool, which are essential oil components of plants, are not only dependent on altitude but also on changes in soil texture and the total CaCO3 content of the soil.
En este estudio, se investigaron los efectos de la altitud y las propiedades del suelo sobre las cantidades de aceite esencial y los componentes del aceite esencial de la planta de salvia turca (Salvia fruticosa Mill). Se definió como área de estudio una ubicación de 300-500 m a 600-800 m en el sur de Turquía en el distrito de Kas de Antalya con rangos de altitud de 0-200 m. Durante el período de plena floración de S. fruticosa, se recolectaron plantas y muestras de suelo. El contenido de los aceites esenciales de las plantas se analizó mediante GC-MS. Según los resultados obtenidos, el contenido de aceite esencial de las plantas varió entre el 1,91% y el 5,10%. El componente principal de los aceites esenciales de las plantas recolectadas de todas las áreas de estudio fue el 1,8-cineol, al 44,93%. Se concluyó que las variaciones en las proporciones de los componentes limoneno y linalol, que son componentes del aceite esencial de las plantas, no solo dependen de la altitud sino también de los cambios en la textura del suelo y el contenido total de CaCO3 del suelo.
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Aceites Volátiles/química , Características del Suelo , Salvia/química , Altitud , Presión Atmosférica , TurquíaRESUMEN
Cinnamaldehyde (CNM) is an essential-oil component with reported anti-infective, anti-inflammatory, and healing effects, making it an interesting compound for the treatment of wound infection. Herein, we evaluated the effects of topical administration of CNM in experimental wounds infected by Staphylococcus aureus. Swiss mice (n = 12/group) were randomly allocated into three groups (CON: animals with uninfected lesions; Sa: animals with untreated infected lesions; Sa + CNM: animals with infected wounds and treated with CNM). Excisional lesions (64 mm2) were induced at the dorsal area followed by the addition of S. aureus (80 µL of a 1.5 × 108 CFU/mL bacterial suspension). The wounds were treated with CNM (200 µg/wound/day) or vehicle (2% DMSO) for 10 days. Skin samples were taken on the 3rd or 10th treatment day for quantification of inflammatory mediators, bacterial load, immunophenotyping, and histological analysis. The treatment with CNM improved the healing process and attenuated the severity of skin lesions infected by S. aureus. These effects were associated with significant decreases in bacterial loads in CNM-treated wounds. The levels of neutrophils, TNF-α, IL-6, NO, and VEGF were decreased in the lesions treated with CNM. Taken together, these data provide further evidence of the effectiveness of CNM for the treatment of skin infections.
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Infecciones Estafilocócicas , Infección de Heridas , Ratones , Animales , Staphylococcus aureus , Cicatrización de Heridas , Infecciones Estafilocócicas/tratamiento farmacológico , Infecciones Estafilocócicas/patología , Infección de Heridas/tratamiento farmacológicoRESUMEN
The hydro-distilled essential oils obtained from aerial parts of the wild and cultivated Echinophora tenuifolia subsp. sibthorpiana have been analyzed by GC-MS and screened for antimicrobial activity. In total, 28 compounds representing more than 99% of the oils were identified. α-phellandrene (13.22% and 55.27%), δ-3-carene (49.29% and 4.03%), and methyl eugenol (22.59-25.69%) were found as the main components for the wild and cultivated oils, respectively. EOs of the wild and cultivated plants differed significantly in both the percentage of the main components and antifungal effect. α-phellandrene was more dominant in cultivated plants (55.27%) than in wild ones (13.22%), while δ-3-carene was more abundant in the wild plants (49.29%). In the antifungal assays, both oils displayed moderate to high activity against three phytopathogenic fungi; Fusarium oxysporum, Rhizoctonia solani, and Alternaria alternata.
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Apiaceae , Aceites Volátiles , Aceites Volátiles/farmacología , Aceites Volátiles/química , Antifúngicos/farmacología , Antifúngicos/química , Turquía , Hongos , Apiaceae/químicaRESUMEN
The spread of multidrug resistant (MDR) bacteria and resistance genes along the food chain and the environment has become a global, but silent pandemic. To face this challenge, it is of outmost importance to develop efficient strategies to reduce potential contamination by these agents. In the present study, 30 strains of Enterococcus sp., Staphylococcus sp. and Pseudomonas sp. isolated from various surfaces throughout the meat production chain in a goat and lamb slaughterhouse were characterized as MDR bacteria harboring several antibiotic resistance genes (ARGs). The antimicrobial efficacy of natural essential oil components "EOCs" (carvacrol "CA," cinnamaldehyde "CIN," eugenol "EU," geraniol "GE," limonene "LI" and thymol "TH"), HLE disinfectant solution (3-6% H2O2; 2.2-4.4% lactic acid and 12.5-25 mM EDTA in water) and EDTA was tested against these MDR bacteria. Results showed that Minimum Inhibitory Concentrations (MIC) were compound and strain dependent. In addition, the synergistic effect of these antimicrobials was evaluated at 1/2 MIC. Here our study showed particularly promising results regarding the inhibitory effect at sub-inhibitory concentrations, which were confirmed by the analysis of bacterial growth dynamics over 72 h. Furthermore, the inhibitory effect of EOCs, HLE disinfectant solution and EDTA or their combinations was studied in developing and established biofilms of MDR bacteria obtaining variable results depending on the morphological structure of the tested strain and the phenolic character of the EOCs. Importantly, the combination of EOCs with HLE or EDTA showed particularly positive results given the effective inhibition of biofilm formation. Moreover, the synergistic combinations of EU and HLE/EDTA, TH, CA, GE, LI or CIN + EDTA/HLE caused log reductions in established biofilms of several strains (1-6 log10 CFU) depending on the species and the combination used, with Pseudomonas sp. strains being the most susceptible. Given these results, we propose novel antimicrobial formulations based on the combination of sub-inhibitory concentrations of EOCs and HLE or EDTA as a highly promising alternative to currently used approaches. This novel strategy notably shows great potential to efficiently decrease the emergence and spread of MDR bacteria and ARGs in the food chain and the environment, thus supporting the decrease of resistomes and pathogenesis in clinical and industrial areas while preserving the antibiotic therapeutic action.
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Essential oils and their active components have been extensively reported in the literature for their efficient antimicrobial, antioxidant and antifungal properties. However, the sensitivity of these volatile compounds towards heat, oxygen and light limits their usage in real food packaging applications. The encapsulation of these compounds into inorganic nanocarriers, such as nanoclays, has been shown to prolong the release and protect the compounds from harsh processing conditions. Nevertheless, these systems have limited shelf stability, and the release is of limited control. Thus, this study presents a mesoporous silica nanocarrier with a high surface area and well-ordered protective pore structure for loading large amounts of natural active compounds (up to 500 mg/g). The presented loaded nanocarriers are shelf-stable with a very slow initial release which levels out at 50% retention of the encapsulated compounds after 2 months. By the addition of simulated drip-loss from chicken, the release of the compounds is activated and gives an antimicrobial effect, which is demonstrated on the foodborne spoilage bacteria Brochothrixthermosphacta and the potentially pathogenic bacteria Escherichia coli. When the release of the active compounds is activated, a ≥4-log reduction in the growth of B. thermosphacta and a 2-log reduction of E. coli is obtained, after only one hour of incubation. During the same one-hour incubation period the dry nanocarriers gave a negligible inhibitory effect. By using the proposed nanocarrier system, which is activated by the food product itself, increased availability of the natural antimicrobial compounds is expected, with a subsequent controlled antimicrobial effect.
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Antiinfecciosos , Aceites Volátiles , Antibacterianos/química , Antibacterianos/farmacología , Antiinfecciosos/análisis , Antiinfecciosos/farmacología , Preparaciones de Acción Retardada/farmacología , Escherichia coli , Embalaje de Alimentos , Aceites Volátiles/química , Aceites Volátiles/farmacología , Dióxido de SilicioRESUMEN
The toxicological properties of different silica particles functionalised with essential oil components (EOCs) were herein assessed using the in vivo model C. elegans. In particular, the effects of the acute and long-term exposure to three silica particle types (SAS, MCM-41 micro, MCM-41 nano), either bare or functionalised with eugenol or vanillin, were evaluated on different biological parameters of nematodes. Acute exposure to the different particles did not reduce nematodes survival, brood growth or locomotion, but reproduction was impaired by all the materials, except for vanillin-functionalised MCM-41 nano. Moreover, long-term exposure to particles led to strongly inhibited nematodes growth and reproduction. The eugenol-functionalised particles exhibited higher functionalisation yields and had the strongest effects during acute and long-term exposures. Overall, the vanillin-functionalised particles displayed milder acute toxic effects on reproduction than pristine materials, but severer toxicological responses for the 96-hour exposure assays. Our findings suggest that the EOC type anchored to silica surfaces and functionalisation yield are crucial for determining the toxicological effects of particles on C. elegans. The results obtained with this alternative in vivo model can help to anticipate potential toxic responses to these new materials for human health and the environment.
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Caenorhabditis elegans , Eugenol , Animales , Benzaldehídos/toxicidad , Eugenol/toxicidad , Humanos , Dióxido de Silicio/toxicidadRESUMEN
The cytotoxicity of carvacrol- and thymol-functionalised mesoporous silica microparticles (MCM-41) was assessed in the human hepatocarcinoma cell line (HepG2). Cell viability, lactate dehydrogenase (LDH) activity, reactive oxygen species (ROS) production, mitochondrial membrane potential (ΔΨm), lipid peroxidation (LPO) and apoptosis/necrosis analyses were used as endpoints. The results showed that both materials induced cytotoxicity in a time- and concentration-dependent manner, and were more cytotoxic than free essential oil components and bare MCM-41. This effect was caused by cell-particle interactions and not by degradation products released to the culture media, as demonstrated in the extract dilution assays. LDH release was a less sensitive endpoint than the MTT (thiazolyl blue tetrazolium bromide) assay, which suggests the impairment of the mitochondrial function as the primary cytotoxic mechanism. In vitro tests on specialised cell functions showed that exposure to sublethal concentrations of these materials did not induce ROS formation during 2 h of exposure, but produced LPO and ΔΨm alterations in a concentration-dependent manner when cells were exposed for 24 h. The obtained results generally support the hypothesis that the carvacrol- and thymol-functionalised MCM-41 microparticles induced toxicity in HepG2 cells by an oxidative stress-related mechanism that resulted in apoptosis through the mitochondrial pathway.
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Cimenos/toxicidad , Dióxido de Silicio/toxicidad , Timol/toxicidad , Apoptosis/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Cimenos/química , Células Hep G2 , Humanos , Peroxidación de Lípido/efectos de los fármacos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Estrés Oxidativo/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Dióxido de Silicio/química , Timol/química , ToxicologíaRESUMEN
Population growth directly affects the global food supply, demanding a higher production efficiency without farmland expansion - in view of limited land resources and biodiversity loss worldwide. In such scenario, intensive agriculture practices have been widely used. A commonly applied method to maximize yield in animal production is the use of subtherapeutic doses of antibiotics as growth promoters. Because of the strong antibiotic selection pressure generated, the intense use of antibiotic growth promoters (AGP) has been associated to the rise of antimicrobial resistance (AMR). Also, cross-resistance can occur, leading to the emergence of multidrug-resistant pathogens and limiting treatment options in both human and animal health. Thereon, alternatives have been studied to replace AGP in animal production. Among such alternatives, essential oils and essential oil components (EOC) stand out positively from others due to, besides antimicrobial effectiveness, improving zootechnical indexes and modulating genes involved in resistance mechanisms. This review summarizes recent studies in essential oils and EOC for zoonotic bacteria control, providing detailed information about the molecular-level effects of their use in regard to AMR, and identifying important gaps to be filled within the animal production area.
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Antiinfecciosos , Aceites Volátiles , Animales , Antibacterianos/farmacología , Antiinfecciosos/farmacología , Farmacorresistencia Bacteriana , Aceites Volátiles/farmacologíaRESUMEN
Mesoporous silica microparticles functionalized with lactose for the specific release of essential oil components (EOCs) in the small intestine are presented. In vitro and in vivo intestinal models were applied to validate the microparticles (M41-EOC-L), in which the presence of lactase acts as the triggering stimulus for the controlled release of EOCs. Among the different microdevices prepared (containing thymol, eugenol and cinnamaldehyde), the one loaded with cinnamaldehyde showed the most significant Caco-2 cell viability reduction. On the other hand, interaction of the particles with enterocyte-like monolayers showed a reduction of EOCs permeability when protected into the designed microdevices. Then, a microdevice loaded with cinnamaldehyde was applied in the in vivo model of Wistar rat. The results showed a reduction in cinnamaldehyde plasma levels and an increase in its concentration in the lumen of the gastrointestinal tract (GIT). The absence of payload release in the stomach, the progressive release throughout the intestine and the prolonged stay of the payload in the GIT-lumen increased the bioavailability of the encapsulated compound at the site of the desired action. These innovative results, based on the specific intestinal controlled delivery, suggest that the M41-payload-L could be a potential hybrid microdevice for the protection and administration of bioactive molecules in the small intestine and colon.
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Encapsulation in liposomes has been an efficient strategy to improve the stability of sensitive bioactive compounds such as essential oils (EOs). However, the stability of liposomal formulations remains a key parameter controlling the delivery of encapsulated ingredients. Cholesterol (Chol) modulates the membrane properties conferring stability to the lipid bilayer. Thus, the Chol content in the liposome formulations encapsulating EO components should be carefully chosen. In this work, various liposome formulations differing by Chol content (DPPC:Chol 100:10; 100:25; 100:50; 100:75; 100:100) were exposed to a series of 22 EO components at DPPC/EO 100/25. The formulations were characterized for their final composition and their permeability to the hydrophilic fluorophore, sulforhodamine B (SRB), was monitored. Results showed that the Chol content experimentally determined for the various formulations (above 10% Chol) was below the theoretical weighed Chol. Among the tested components, 13 molecules displayed a significant permeabilizing effect on 10% Chol membranes. Most of these possess a hydroxyl group. The EO induced permeability was dependent on the Chol content which affects the membrane phase: their effect was reduced upon increasing Chol content keeping five EOs components effective at 40% Chol. The EO's effect was also linked to the hydrophobicity of the molecule. Hence, the DPPC:Chol ratio of the formulation is chosen considering the structure of the compound, its hydrophobicity and its effect on the permeability at different Chol content: a formulation comprising 40% Chol is suggested for highly hydrophobic molecules whereas a formulation with higher Chol content could be selected for less hydrophobic compounds.
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Liposomas , Aceites Volátiles , 1,2-Dipalmitoilfosfatidilcolina/química , Colesterol/química , Membrana Dobles de Lípidos , Liposomas/química , PermeabilidadRESUMEN
The development of new food preservatives is essential to prevent foodborne outbreaks or food spoilage due to microbial growth, enzymatic activity or oxidation. Furthermore, new compounds that substitute the commonly used synthetic food preservatives are needed to stifle the rising problem of microbial resistance. In this scenario, we report herein, as far as we know, for the first time the use of the zein protein as a gating moiety and its application for the controlled release of essential oil components (EOCs). The design of microdevices consist of mesoporous silica particles loaded with essential oils components (thymol, carvacrol and cinnamaldehyde) and functionalized with the zein (prolamin) protein found in corn as a molecular gate. The zein protein grafted on the synthesized microdevices is degraded by the proteolytic action of bacterial enzymatic secretions with the consequent release of the loaded essential oil components efficiently inhibiting bacterial growth. The results allow us to conclude that the new microdevice presented here loaded with the essential oil component cinnamaldehyde improved the antimicrobial properties of the free compound by decreasing volatility and increasing local concentration.
Asunto(s)
Antibacterianos/química , Aceites Volátiles/química , Dióxido de Silicio/química , Zeína/química , PorosidadRESUMEN
In time, diagnosis and detection of virulence factor and its pathogenomics study continues to grow and this leads to novel treatments for infectious diseases. The objective of this study was to detect and characterise virulence genes in a haemolytic strain of Staphylococcus aureus in vitro and molecular interaction studies with herbal essential oil components in silico. A hospital biosample-isolated strain of Staphylococcus aureus (BMS-2) was resistant towards Cephalosporin. The PCR-amplified FASTA nucleotide sequence was identical with S. aureus strains absolutely. The calculated GC value was 34.05%. The translated protein sequence was identified with a conserved domain of hlyII ß-channel forming cytolysin belonging to leukocidin superfamily and was predicted as a stable, non-transmembrane protein comprising B cell epitopes. Structurally, the protein was found to be composed of α helix, π-helix, extended strands, ß-sheet, turn and bends with atomic composition as C658H1026N174O200S2. The molecular docking studies made between the HlyII cytolysin (receptor) and wet lab studied essential oil components (citral a, citronellol, eucalyptol, eugenol, geraniol, linalool, menthol, piperine and thymol) as ligands using Autodock 1.5.6 tool had inferred about prevalence of hydrogen bonds as well as covalent bonds in the intermolecular interactions. Amino acids like Tyr68, Tyr 69, Asn106, Asp67 and Asn106 were observed to be the most active residues for H-bond and hydrophobic bonds respectively. Only geraniol had interaction with glycine residue of the toxin molecule. In conclusion, geraniol with the highest ligand efficiency was observed to be the most potent phyto-constituent interacting with the in vitro detected hlyII cytotoxin.
Asunto(s)
Proteínas Bacterianas/química , Simulación del Acoplamiento Molecular , Aceites Volátiles/química , Perforina/química , Staphylococcus aureus/química , Proteínas Bacterianas/antagonistas & inhibidores , Proteínas Bacterianas/genética , Humanos , Perforina/antagonistas & inhibidores , Perforina/genética , Estructura Secundaria de Proteína , Staphylococcus aureus/genética , Staphylococcus aureus/patogenicidadRESUMEN
This study aimed to evaluate possible synergistic interactions on antimicrobial and antioxidant efficacy of clove and cinnamon oil components in combination and characterization of compounds responsible for synergistic interactions using TLC bioautography followed by checkerboard titration, isobologram analysis, and spectrometric characterization. Among the combinations tested, cinnamaldehyde from cinnamon oil and eugenol from clove oil in combination showed a synergistic antimicrobial interaction against foodborne microbes Listeria monocytogenes (fractional inhibitory concentration index (FICI): 0.31), Salmonella typhimurium (FICI: 0.41), and Aspergillus niger (FICI: 0.48), and synergistic antioxidant efficacy (combination index: 0.78) in in vitro model. Cinnamaldehyde/eugenol blend did not show any cytotoxic effect (IC50 > 1000 µg/ml) in human normal keratinocyte cell line. The results provide evidence that the cinnamaldehyde/eugenol blend may help in designing a more potent novel natural antimicrobial and antioxidant agent in food and pharmaceutical industries.