Avaliação da qualidade e vida de prateleira de ovos de consumo tratados com óleo mineral e timol / Evaluation of the quality and shelf life of consumer eggs treated with mineral oil and thymol

Várias pesquisas mostram o interesse na aplicação de materiais de revestimento na superfície de cascas de ovos para preservar a qualidade interior dos ovos, bem como para prolongar sua vida útil. No presente trabalho verificou-se a qualidade interna de 100 ovos galinhas para o consumo. Os ovos foram revestidos externamente na casca com tratamento de óleo mineral e timol 10%, e um grupo controle, que não sofreu nenhum tipo de tratamento. Um total de 10 ovos de cada grupo (controle e tratado) foram avaliados no dia 7, 14, 21 dias de armazenamento em relação aos seus parâmetros internos de qualidade. Os parâmetros físico-químicos de qualidade dos ovos avaliados foram Perda de peso dos ovos (g), Porcentagem da gema (%), Porcentagem do albúmen (%), Índice de gema, pH do Albúmen, pH da gema e Unidades Haugh (UH). Foi observada diminuição da qualidade interna dos ovos durante os períodos de armazenamento principalmente no grupo controle. Os ovos do grupo controle, que não receberam nenhum tipo de revestimento apresentaram a maior perda de peso e os valores da unidade Haugh diminuíram ao longo do armazenamento, o que não foi observado nos dois grupos tratados. A unidade Haugh dos grupos tratados não apresentaram diferenças significativas entre si. Concluiu-se que os ovos tratados com óleo mineral e timol a 10%, o revestimento da casca contribuiu para a preservação dos ovos mantendo positivamente a qualidade interna dos ovos ao longo do período de estocagem, o que pode contribuir para o aumento da vida de prateleira de ovos.

Several researchers show interest in the application of coating materials on the surface of eggshells to preserve the interior quality of the eggs, as well as to prolong their shelf life. In the present work, the internal quality of 100 chicken eggs for consumption was verified. The eggs were coated externally in the shell with mineral oil and 10% thymol treatment, and a control group did not undergo any type of treatment. a total of 10 eggs from each group (control and treated) were evaluated on day 7, 14, 21 of storage in relation to their internal quality parameters. The physicochemical parameters of egg quality evaluated were egg weight loss (g), yolk percentage (%), albumen percentage (%), yolk index, albumen pH, yolk ph and haugh units (uh). decreased internal egg quality was observed during storage periods, mainly in the control group. The eggs in the control group, which did not receive any kind of coating, showed the greatest weight loss and the haugh unit values decreased during storage, which was not observed in the two treated groups. The haugh unit of the treated groups did not show significant differences between them. It was concluded that in eggs treated with mineral oil and 10% thymol, the shell coating contributed to the preservation of the eggs, positively maintaining the internal quality of the eggs throughout the storage period, which may contribute to an increase in shelf life of eggs.

Estudo do potencial antifúngico e do mecanismo de ação do timol contra cepas de Candida parapsilosis resistentes ao fluconazol e a anfotericina B / Study of the antifungal potential and mechanism of action of thymol against strains of Candida parapsilosis resistant to fluconazole and amphotericin B

Objetivo: investigar a suscetibilidade de cepas fúngicas de Candida parapsilosis isoladas de sangue humano frente ao timol, bem como seu mecanismo de ação. Metodologia: foram utilizadas técnicas de microdiluição em placas de 96 poços para determinar a concentração inibitória mínima (CIM) e concentração fungicida mínima (CFM). Além disso, foram realizados testes com o sorbitol e o ergosterol para investigar a ação do timol na parede e na membrana celular fúngica respectivamente. Resultados: nos testes de CIM e CFM, foi observado que as cepas de C. parapsilosis são resistentes ao fluconazol e a anfotericina B, no entanto, o timol desempenhou efeito fungicida com razão CFM/CIM entre 1 e 2. Além disso, a CIM do timol não aumentou quando o sorbitol ou o ergosterol foi adicionado no meio, sugerindo fortemente que este monoterpeno não age na parede celular fúngica ou por ligação ao ergosterol na membrana plasmática. Conclusão: portanto, esses resultados contribuem para a elucidação do mecanismo de ação do timol, sugerindo outros possíveis alvos de interação fármaco-receptor. No entanto, mais investigações de caráter enzimático e molecular em modelos in vitro são necessários para que se possa elucidar completamente o modo de ação desse promissor monoterpeno.

Objective: to investigate the susceptibility of fungal strains of Candida parapsilosis isolated from human blood against thymol, as well as its mechanism of action. Methodology: microdilution techniques were used in 96-well plates to determine minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC). In addition, tests were performed with sorbitol and ergosterol to investigate the action of thymol on the wall and on the fungal cell membrane respectively. Results: in the CIM and CFM tests, it was observed that C. parapsilosis strains are resistant to fluconazole and amphotericin B, however, thymol had a fungicidal effect with MFC/MIC ratio between 1 and 2. In addition, thymol MIC did not increase when sorbitol or ergosterol was added in the medium, strongly suggesting that this monoterpene does not act on the fungal cell wall or by binding to ergosterol on the plasma membrane. Conclusion: therefore, these results contribute to the elucidation of the mechanism of action of thymol, suggesting other possible targets of drug-receptor interaction. However, further investigations of enzymatic and molecular character in in vitro models are necessary to fully elucidate the mode of action of this promising monoterpene.

Effect of thymol and environmental factors on growth and biofilm formation by Listeria monocytogenes / Efecto del timol y factores ambientales sobre el crecimiento y formación de biopelícula por Listeria monocytogenes

Thymol (2-isopropyl-5-methylphenol) is an aromatic monoterpene found in essential oils extracted from plants belonging to the Lamiaceae family, such as Thymus, Ocimum, Origanum, Satureja, Thymbra and Monarda genera. Growth and biofilm formation by Listeria monocytogenes CLIP 74902 were evaluate using three carbon sources in the presence of thymol. Specific growth rate (h-1) values at 37o with glucose, trehalose and cellobiose with the addition of thymol (µg/mL) 0 (control) and 750, were respectively: 0.22, 0.07; 0.14, 0.04; 0.11, 0.04. Lag periods obtained under the same conditions were (h): 8.19, 13.2; 22.5, 27.5; 23.1, 28.1. A marked antibiofilm activity was observed against the exposure with 750 µg/mL of thymol, showing a high percentage of inhibition: glucose (99 %), trehalose (97 %) and cellobiose (98%), compared to the control. The results suggest that thymol could be used to inhibit the growth and production of biofilms by L. monocytogenes in the food industry.

Timol (2-isopropil-5-metilfenol) es un monoterpeno aromático presente en los aceites esenciales extraídos de plantas pertenecientes a la familia Lamiaceae, como los géneros Thymus, Ocimum, Origanum, Satureja, Thymbra y Monarda. El crecimiento y formación de biopelícula por Listeria monocytogenes CLIP 74902 fueron evaluados utilizando tres fuentes de carbono en presencia de timol. La velocidad específica de crecimiento (h-1) a 37o con glucosa, trehalosa y celobiosa con la adición de timol (µg/mL) 0 (control) y 750, fueron respectivamente: 0.22, 0.07; 0.14, 0.04, 0.11, 0,04. Los períodos lag obtenidos en las mismas condiciones fueron (h): 8.19, 13.2; 22.5, 27.5; 23.1, 28.1. Una marcada actividad antibiofilm fue obtenida con 750 µg/mL de timol, mostrando un alto porcentaje de inhibición con glucosa (99%), trehalosa (97%) y celobiosa (98%), respecto al control. Los resultados sugieren que timol podría ser usado para inhibir el crecimiento y producción de biopelículas por L. monocytogenes en la industria alimentaria.

Essential oil composition in natural population of Lippia origanoides (Verbenaceae) during dry and rainy seasons / Composición de los aceites esenciales en poblaciones naturales de Lippia origanoides (Verbenaceae) durante las estaciones seca y lluviosa

Abstract Lippia origanoides Kunth. is a medicinal shrub native to some countries in South America, Central America and the Caribbean. This study aims to investigate the chemical composition of the essential oil and the genetic diversity of a natural population of Lippia origanoides in two seasons. Leaf samples were collected from 30 individuals in the dry and rainy seasons. The essential oil was extracted into a Clevenger apparatus and analyzed by gas chromatography coupled to mass spectrometry (GC-MS). The relative quantification of thymol and carvacrol was performed by gas chromatography coupled to a flame ionization detector (GC-FID). Multivariate analysis was used to evaluate the genetic divergence between plants. Carvacrol was the major compound found in most plants for both seasons. In the rainy season, plant 15 presented thymol as the major compound, and plants 16, 27, 28 and 29 presented camphor as the major compound. In the dry season, thymol remained the main compound of plant 15 and camphor remained the main compound of plants 16, 28 and 29; however, plant 27 presented carvacrol as the main compound in this season. After carvacrol, the compounds with the highest content were γ-terpinene, p-cymene and methyl-ether-thymol. Of the 30 plants studied, only five differed in their chemical composition, showing some degree of stability in relation to the dry and rainy seasons. There was no variation in the main compounds between the two seasons, but there was chemical diversity among the main compounds. Tocher grouping revealed five distinct groups, with group 1 including most of the plants in both dry and rainy seasons, indicating that the time of year evaluated did not significantly interfere with the essential oil composition of most plants.(AU)

Resumen Lippia origanoides Kunth. es un arbusto medicinal nativo de algunos países de Centro y Sur América y el Caribe. El objetivo de este estudio fue investigar la composición química de los aceites esenciales y la diversidad genética de una población natural de L. origanoides en dos estaciones. El aceite esencial se extrajo en un equipo de Clevenger y se analizó por cromatografía de gases acoplado a espectrometría de masas (CG-EM). La cuantificación relativa de timol y carvacrol se realizó por cromatografía de gases acoplado a un detector de ionización de llamas (CG-DIL). Se utilizó un análisis multivariado para evaluar la divergencia genética entre plantas. El carvacrol fue el compuesto principal encontrado en la mayoría de las plantas para ambas estaciones. En la estación lluviosa, la planta 15 presentó timol como el compuesto principal y las plantas 16, 27, 28 y 29 presentaron alcanfor como el compuesto principal. En la estación seca, el timol se mantuvo como el principal compuesto en la planta 15, de igual manera el alcanfor se mantuvo como el principal en las plantas 16, 28 y 29. Sin embargo, la planta 27 presentó carvacrol como el principal compuesto en esta estación. Después del carvacrol, los compuestos con el mayor contenido fueron γ-terpineno, p-cimeno and metil-eter-timol. De las 30 plantas estudiadas, solo cinco difirieron en su composición química, mostrando algún grado de estabilidad en relación a la estación seca y lluviosa. No hubo variación en los compuestos principales entre las dos estaciones, pero hubo diversidad química entre los compuestos principales. El agrupamiento de Tocher mostró cinco grupos distintos, con el grupo 1 incluyendo la mayoría de las plantas en ambas estaciones, lo cual indica que el periodo del año evaluado no interfiere significativamente con la composición de aceites esenciales en la mayoría de plantas.(AU)

Thymol Rich Thymbra capitata Essential Oil Inhibits Quorum Sensing, Virulence and Biofilm Formation of Beta Lactamase Producing Pseudomonas aeruginosa

Infections with Pseudomonas aeruginosa are difficult to treat not only because it is often associated with multidrug-resistant infections but also it is able to form biofilm. The aim of this study was to evaluate the antibiofilm and anti-Quorum Sensing (QS) activities of Thymbra capitata essential oils (EOs) against Beta Lactamase (BL) producing P. aeruginosa and the reference strain P. aeruginosa 10145. GC/MS analysis showed that thymol (23.25%) is the most dominant compound in T. capitata EOs. The MICs of T. capitata EOs against P. aeruginosa (BL) and P. aeruginosa 10145 were 1.11%. At sub MIC (0.041, 0.014 and 0.0046%), the EOs of T. capitata remarkably inhibited the biofilm formation of both strains tested and complete inhibition of the biofilm formation was reported at 0.041%. The EOs of T. capitata were found to inhibit the swarming motility, aggregation ability and hydrophobic ability of P. aeruginosa (BL) and P. aeruginosa 10145. Interestingly, the EOs of T. capitata reduce the production of three secreted virulence factors that regulated by QS system including pyocyanin, rhamnolipids and LasA protease. The potent antibiofilm and anti-QS activities of T. capitata EOs can propose it as a new antibacterial agent to control pseudomonas infections.

Avaliação do efeito de soluções de armazenamento sobre a resistência da união adesiva à dentina de dentes bovinos sob microcisalhamento - pesquisa in vitro / Evaluation of the effect of storage solutions on the bond strength to the dentin of bovine teeth under micro-incision - in vitro research

Introdução: Com a dificuldade de obtenção de dentes humanos para estudos in vitro, autores relatam e preconizam o uso de dentes extraídos de arcadas bovinas para testar técnicas de diferentes segmentos da Odontologia. Entretanto, para que as pesquisas tenham eficácia científica, estes dentes necessitam tratamento adequado para manter suas características e simular condições in vivo. Objetivo: Analisar a resistência de união (RU) adesiva à dentina em dentes bovinos restaurados com adesivo convencional Adper® Single Bond 2 (3M ESPE, St. Paul, MN, EUA) e compósito Opallis Flow (Dentscare LTDA., Joinville, SC, BRA), tendo como fatores de estudo solução e tempo de armazenamento. Metodologia: As amostras foram congeladas sob armazenamento em 2 soluções: timol 0,1% e cloramina 0,5%, em 4 diferentes períodos: 1 semana, 1 mês, 3 meses e 6 meses. Foram obtidos 8 grupos experimentais (n=64) e 1 grupo controle (n=8), totalizando 9 grupos. Após o período de restauração, os dentes foram armazenados em estufa a 37 ºC, imersos em água destilada e, em seguida, testados em máquina de ensaios universal (EMIC DL 500, São José dos Pinhais, PR, Brasil), com análise estatística dos resultados. Resultado: A comparação entre os grupos experimentais e controle mostrou diferença estatisticamente significante (p<0,05) para o armazenamento das amostras em timol durante o período de 1 mês. Conclusão: Os fatores solução e tempo foram capazes de alterar a resistência de união adesiva à dentina bovina, observando-se que a maior média de RU foi obtida após o armazenamento na solução timol durante 1 mês (p=0,004).

Introduction: With the difficulty of obtaining human teeth for in vitro studies, authors report and recommend the use of teeth extracted from bovine arches to test techniques from different segments of Dentistry. However, for research to be scientifically effective, these teeth need adequate treatment to maintain their characteristics and to mimic conditions in vivo. Objective: Analyze the adhesive bond strength (RU) to dentin in bovine teeth restored with Adper® Single Bond 2 (3M ESPE, St Paul, MN, USA) and Opallis Flow composite adhesive (Dentscare LTDA, Joinville, SC, BRA), taking as study factors solution and storage time. Methodology: The samples were frozen under storage in 2 solutions: 0.1% thymol and 0.5% chloramine, in 4 different periods: 1 week, 1 month, 3 months and 6 months. Eight experimental groups (n = 64) and one control group (n = 8) were obtained, totaling 9 groups. After the restoration period, the teeth were stored in an incubator at 37ºC, immersed in distilled water and then tested in a universal testing machine (EMIC DL 500, São José dos Pinhais, PR, Brazil), with statistical analysis of the results. Result: The comparison between the experimental and control groups showed a statistically significant difference (p <0.05) for the storage of thymol samples during the 1 month period. Conclusion: Solution and time factors were able to change the adhesive bond strength to the bovine dentin, observing that the highest mean of RU was obtained after storage in the thymol solution for 1 month (p = 0.004).

Susceptibility of cariogenic microorganisms to phytoconstituents / Susceptibilidade de microrganismos cariogênicos a fitoconstituintes

Abstract This study aimed to evaluate the in vitro antibacterial activity of the phytochemicals thymol, linalool, and citronellol against Streptococcus mutans, Streptococcus salivarius and Streptococcus oralis. Disk diffusion screening on solid medium and measurement of the diameter of the bacterial growth inhibition halos was the technique utilized. The Minimum Inhibitory Concentration (MIC) of the substances was determined using serial substance dilutions and microdilution technique in Brain Heart Infusion culture medium. After incubation for 24 hours in an oven at 37 °C, plate reading was completed and confirmed by visual method using 2,3,5 triphenyl tetrazolium chloride dye. The Minimum Bactericidal Concentration (MBC) was determined from MIC subcultures. Assays were performed in triplicate, and chlorhexidine was used as a positive control. The diameters in mm of the growth inhibition halos ranged between 7.3 and 10.7 for S. mutans, 7.3 and 10.0 for S. oralis, and 8.2 and 9.8 for S. salivarius. The MIC and MBC values obtained converged, ranging from maximum values in the presence of Linalool (1,250.0 mg/mL, 2,500.0 mg/mL and 2,500.0 mg/mL, respectively, for S. mutans, S. oralis, and S. salivarius); and minimum values with Thymol (312.5 μg/ml, 156.2 μg/mL and 156.2 μg/ml, respectively for S. mutans, S. oralis, and S. salivarius). All the tested phytochemicals displayed antibacterial activity, thus representing substances with potential applications in preventing tooth decay.

Resumo Este estudo objetivou avaliar a atividade antibacteriana in vitro dos fitoquímicos timol, linalol e citronelol sobre Streptococcus mutans, Streptococcus salivaris e Streptococcus oralis. Utilizou-se a técnica de discos de difusão em meio sólido e medição do diâmetro dos halos de inibição. A concentração inibitória mínima (CIM) das substâncias foi determinada utilizando diluições em série das substâncias e técnica de microdiluição em meio de cultura de Brain Heart Infusion. Após incubação durante 24 horas em estufa a 37 °C, a leitura da placa foi confirmada pelo método visual usando o corante 2,3,5 trifenil cloreto de tetrazólio. A concentração bactericida mínima (CBM) foi determinada a partir de subculturas de MIC. Os ensaios foram realizados em triplicata, e clorexidina foi usada como um controle positivo. Os diâmetros dos halos de inibição do crescimento variaram entre 7,3 e 10,7 por S. mutans, 7,3 e 10,0 por S. oralis, e 8,2 e 9,8 para S. salivaris. Os valores de CIM e CBM obtidos variaram de valores máximos na presença de linalol (1.250,0 mg/mL, 2.500.0 mg/mL e 2.500.0 mg/mL, respectivamente, para o S. mutans, S oralis e S. salivaris); a valores mínimos com timol (312,5 μg/ml, 156,2 μg/mL e 156,2 μg/ml, respectivamente para S. mutans, S. oralis e S. salivaris). Todos os fitoquímicos testados apresentaram atividade antibacteriana, representando, assim, substâncias com potencial de aplicações na prevenção da cárie dentária.

Proliferation of human adipose tissue-derived stem cells stimulated by oil rich in thymol of Lippia origanoides

Abstract Purpose: To evaluate the effects of this thymol-rich oil in the proliferation of human adipose tissue-derived stem cells. Methods: Stem cells were isolated from human adipose tissue by liposuction. After the first passage, cells were cultivated in triplicate for three days in control medium and medium supplemented with three oil samples (1.0 μg/mL, 5.0 μg/mL, and 25.0 μg/mL). Cells were analyzed by the MTT assay at passage 1 (P1), and cell proliferation of control and 1 μg/mL groups was determined with a hemocytometer at P2 and P3. Results: Viability of the essential oil-treated cells was significantly higher than the control group at P1 (p = 0.0008). The treatment with the oil, at a concentration of 1 µg/mL, led to increases of 24.8% at P1 and 43.0% at P3 in the rate of cell proliferation compared with control cells. Conclusion: Supplementing culture medium with essential oil of Lippia origanoides increased cell proliferation, especially at later passages.

Growth modeling to control (in vitro) Fusarium verticillioides and Rhizopus stolonifer with thymol and carvacrol / Modelización del crecimiento in vitro para controlar Fusarium verticillioides y Rhizopus stolonifer con timol y carvacrol

The aim of this study was to evaluate the antifungal activity (in vitro) of thymol and carvacrol alone or in mixtures against Fusarium verticillioides and Rhizopus stolonifer, and to obtain primary growth models. Minimal inhibitory concentration (MIC) was evaluated with fungal radial growth with thymol or carvacrol concentrations (0-1600mg/l). Mixtures were evaluated using concentrations below MIC values. Radial growth curves were described by the modified Gompertz equation. MIC values of carvacrol were 200mg/l for both fungi. Meanwhile, MIC values of thymol were between 500 and 400mg/l for F verticillioides and R. stolonifer, respectively. A synergistic effect below MIC concentrations for carvacrol (100mg/l) and thymol (100-375 mg/l) was observed. Significant differences (p <0.05) between the Gompertz parameters for the antimicrobial concentrations and their tested mixtures established an inverse relationship between antimicrobial concentration and mycelial development of both fungi. Modified Gompertz parameters can be useful to determine fungistatic concentrations.

El objetivo de este trabajo fue evaluar la actividad antifúngica in vitro del timol y del carvacrol, solos o en mezclas, contra Fusarium verticillioides y Rhizopus stolonifer, y obtener modelos primarios de crecimiento. Se evaluó la concentración inhibitoria mínima (CIM) con el crecimiento radial, se ensayaron concentraciones de timol y carvacrol de 0 a 1.600 mg/l. Las mezclas se evaluaron utilizando concentraciones por debajo de los valores de CIM. Las curvas de crecimiento radial fueron descritas por la ecuación de Gompertz modificada. Se obtuvieron los siguientes valores de CIM: carvacrol, 200 mg/l para las 2 especies; timol, 500 mg/l y 400 mg/l para F. verticillioides y R. stolonifer, respectivamente. Se observó un efecto sinèrgico a concentraciones inferiores a las CIM para el carvacrol (100mg/l) y el timol (100-375 mg/l). Hubo diferencias significativas (p <0,05) entre los parámetros de crecimiento de Gompertz; se estableció que existe una relación inversa entre la concentración de los antimicrobianos y el desarrollo del micelio de ambos hongos.

The Major Postharvest Disease of Onion and Its Control with Thymol Fumigation During Low-Temperature Storage

Onion (Allium cepa L.) is one of the major vegetable crops in Korea that are damaged and lost by pathogenic fungal infection during storage due to a lack of proper storage conditions. The aim of this study was to determine an appropriate control measure using thymol to increase the shelf life of onions. To control fungal infections that occur during low-temperature storage, it is necessary to identify the predominant fungal pathogens that appear in low-temperature storage houses. Botrytis aclada was found to be the most predominant fungal pathogen during low-temperature storage. The antifungal activity of the plant essential oil thymol was tested and compared to that of the existing sulfur treatments. B. aclada growth was significantly inhibited up to 16 weeks with spray treatments using a thymol solution. To identify an appropriate method for treating onions in a low-temperature storage house, thymol was delivered by two fumigation treatment methods, either by heating it in the granule form or as a solution at low-temperature storage conditions (in vivo). We confirmed that the disease severity was reduced up to 96% by fumigating thymol solution compared to the untreated control. The efficacy of the fumigation of thymol solution was validated by testing onions in a low-temperature storage house in Muan, Jeollanam-do. Based on these results, the present study suggests that fumigation of the thymol solution as a natural preservative and fungicide can be used as an eco-friendly substitute for existing methods to control postharvest disease in long-term storage crops on a commercial scale.

Efecto antimicrobiano del cinamaldehído, timol, eugenol y quitosano sobre cepas de Streptococcus mutans / Antimicrobial effect of cinnamaldehyde, thymol, eugenol and chitosan on Streptococcus mutans strains

Objetivo: evaluar la actividad antimicrobiana de cinamaldehído, timol, eugenol y quitosano en comparación con clorhexidina al 0,12 por ciento sobre cepas de Streptococccus mutans. Métodos: se realizaron pruebas de susceptibilidad bacteriana de cepas de Streptococccus mutans con discos embebidos en eugenol, cinamaldehído, quitosano y timol, al 0,1 y 1 por ciento con un control positivo de clorhexidina al 0,12 por ciento evaluando los halos de inhibición existentes a las 24 h y 48 h. Resultados: el cinamaldehído al 1 por ciento mostró el mejor poder de control sobre el crecimiento de Streptococccus mutans al presentar 19,91 mm y 24,44 mm de halos de inhibición a las 24 h y 48 h, respectivamente, con una diferencia significativa de p< 0,05 entre los dos intervalos. Cuando se comparó el cinamaldehído al 1 por ciento con el control positivo de clorhexidina al 0,12 por ciento, no se encontraron diferencias significativas (p> 0,05). Conclusiones: el cinamaldehído al 1 por ciento tuvo los mejores resultados seguidos por quitosano, eugenol y timol a la misma concentración en comparación con clorhexidina al 0,12 por ciento tanto a la 24 como 48 h(AU)

Objective: to evaluate the antimicrobial activity of cinnamaldehyde, thymol, eugenol and chitosan compared to chlorhexidine-0.12 percent on Streptococcus mutans strains. Methods: bacterial susceptibility tests were performed on S. mutans strains with disks embedded in eugenol, cinnamaldehyde, chitosan and thymol, at 0.1 percent and 1 percent with a positive control of 0.12 percent chlorhexidine, evaluating existing inhibition zones after 24 and 48 hours. Results: cinnamaldehyde-1 percent showed the best control power over the growth of S. mutans, presenting 19.91 mm and 24.44 mm of inhibition halos after 24 and 48 hours, respectively, with a significant difference of p< 0.05 between the two intervals. When the cinnamaldehyde-1 percent was compared with the positive control of chlorhexidine-0.12 percent, no significant differences were found (p> 0.05). Conclusions: cinnamaldehyde-1 percent had the best results followed by chitosan, eugenol and thymol in the same concentration compared to chlorhexidine-0.12 percent after both 24 and 48 hours(AU)

Anthelmintic effect of thymol and thymol acetate on sheep gastrointestinal nematodes and their toxicity in mice / Efeito anti-helmíntico do timol e acetato de timila sobre nematoides gastrintestinais de ovinos e toxicidade em camundongos

Abstract Thymol is a monoterpene and acetylation form of this compound can reduce the toxicity and enhance its biological effects. The objective of this study was to evaluate the effect of thymol and thymol acetate (TA) on egg, larva and adult Haemonchus contortus and the cuticular changes, acute toxicity in mice and the efficacy on sheep gastrointestinal nematodes. In vitro tests results were analyzed by analysis of variance (ANOVA) and followed by comparison with Tukey test or Bonferroni. The efficacy of in vivo test was calculated by the BootStreet program. In the egg hatch test (EHT), thymol (0.5 mg/mL) and TA (4 mg/mL) inhibited larval hatching by 98% and 67.1%, respectively. Thymol and TA (8 mg/mL) inhibited 100% of larval development. Thymol and TA (800 µg/mL) reduced the motility of adult worms, by 100% and 83.4%, respectively. Thymol caused cuticular changes in adult worm teguments. In the acute toxicity test, the LD50 of thymol and TA were 1,350.9 mg/kg and 4,144.4 mg/kg, respectively. Thymol and TA reduced sheep egg count per gram of faeces (epg) by 59.8% and 76.2%, respectively. In in vitro tests thymol presented better anthelmintic activity than TA. However TA was less toxic and in in vivo test efficacy was similar.

Resumo Timol é um monoterpeno e a acetilação deste composto pode reduzir a toxicidade e potencializar os seus efeitos biológicos. O objetivo deste trabalho foi avaliar o efeito do timol e acetato de timolila (AT) sobre ovos, larvas e adultos de Haemonchus contortus e suas alterações cuticulares, toxicidade aguda em camundongos e a eficácia sobre nematoides gastrintestinais de ovinos. Os resultados dos testes in vitro foram analisados por análise de variância (ANOVA) e comparados pelo testes de Tukey ou Bonferroni. A eficácia do teste de redução da contagem de ovos nas fezes (RCOF) foi calculada pelo programa BootStreet. No teste de inibição da eclosão de ovos (TEO), timol (0,5 mg/mL) e AT (4 mg/mL) inibiram a eclosão das larvas em 98% e 67,1%, respectivamente. Timol e AT (8 mg/mL) inibiram 100% do desenvolvimento larval. Timol e AT (800 μg/mL) reduziram a motilidade dos nematoides adultos, em 100% e 83,4%, respectivamente. O timol provocou alterações cuticulares nos nematoides adultos. No teste de toxicidade aguda, a DL50 do timol e AT foi de 1.350,9 mg/kg e 4.144,4 mg/kg, respectivamente. Timol e AT reduziram a contagem de ovos por gramas de fezes (OPG) dos ovinos em 59,8% e 76,2%, respectivamente. Nos testes in vitro timol apresentou atividade melhor anti-helmíntica do que AT. Entretanto, AT foi menos tóxico do que o timol e no teste in vivo apresentaram eficácia semelhante.

Antifungal activity of synthetic antiseptics and natural compounds against Candida dubliniensis before and after in vitro fluconazole exposure

ABSTRACT INTRODUCTION: This study evaluated the susceptibilities of oral candidiasis-derived Candida albicans, fluconazole-resistant (FR) Candida dubliniensis, and fluconazole-susceptible (FS) C. dubliniensis to synthetic antiseptics [chlorhexidine gluconate (CHX), cetylpyridinium chloride (CPC), and triclosan (TRC)] and natural compounds (carvacrol, eugenol and thymol). METHODS: Susceptibility tests were performed based on the M27-A3 reference method. The fluconazole-resistant C. dubliniensis strains were obtained after prolonged in vitro exposure to increasing fluconazole concentrations. The geometric mean values for minimum inhibitory concentrations and minimum fungicidal concentrations were compared among the groups. RESULTS: Fluconazole-susceptible C. dubliniensis was more sensitive to CPC and TRC than FR C. dubliniensis and C. albicans were. However, eugenol and thymol were more active against FR C. dubliniensis. The fungicidal activities of CHX and TRC were similar for the three groups, and FR C. dubliniensis and C. albicans had similar sensitivities to CPC. CONCLUSIONS: The resistance of C. dubliniensis to fluconazole affects its sensitivity the synthetic antiseptics and natural compounds that were tested.

Effects of Lippia sidoides essential oil, thymol, p-cymene, myrcene and caryophyllene on rat sciatic nerve excitability

Lippia sidoides Cham is a typical herb species of Northeast Brazil with widespread use in folk medicine. The major constituents of the essential oil of L. sidoides (EOLs) are thymol, p-cymene, myrcene, and caryophyllene. Several studies have shown that the EOLs and its constituents have pharmacological effects, including antibacterial, anti-inflammatory, antioxidant and neuroprotective activity. Therefore, this work aimed to investigate the effects of the EOLs and their main constituents on rat sciatic nerve excitability. The sciatic nerves of adult Wistar rats were dissected and mounted in a moist chamber. Nerves were stimulated by square wave pulses, with an amplitude of 40 V, duration of 100 μs to 0.2 Hz. Both EOLs and thymol inhibited compound action potential (CAP) in a concentration-dependent manner. Half maximal inhibitory concentration for CAP peak-to-peak amplitude blockade were 67.85 and 40 µg/mL for EOLs and thymol, respectively. CAP peak-to-peak amplitude was significantly reduced by concentrations ≥60 µg/mL for EOLs and ≥30 µg/mL for thymol. EOLs and thymol in the concentration of 60 µg/mL significantly increased chronaxie and rheobase. The conduction velocities of 1st and 2nd CAP components were also concentration-dependently reduced by EOLs and thymol in the range of 30-100 µg/mL. Differently from EOLs and thymol, p-cymene, myrcene and caryophyllene did not reduce CAP in the higher concentrations of 10 mM. These data demonstrated that EOLs and thymol inhibited neuronal excitability and were promising agents for the development of new drugs for therapeutic use.

Monoterpenoids (thymol, carvacrol and S-(+)-linalool) with anesthetic activity in silver catfish (Rhamdia quelen): evaluation of acetylcholinesterase and GABAergic activity

This study evaluated the anesthetic potential of thymol and carvacrol, and their influence on acetylcholinesterase (AChE) activity in the muscle and brain of silver catfish (Rhamdia quelen). The AChE activity of S-(+)-linalool was also evaluated. We subsequently assessed the effects of thymol and S-(+)-linalool on the GABAergic system. Fish were exposed to thymol and carvacrol (25, 50, 75, and 100 mg/L) to evaluate time for anesthesia and recovery. Both compounds induced sedation at 25 mg/L and anesthesia with 50-100 mg/L. However, fish exposed to carvacrol presented strong muscle contractions and mortality. AChE activity was increased in the brain of fish at 50 mg/L carvacrol and 100 mg/L thymol, and decreased in the muscle at 100 mg/L carvacrol. S-(+)-linalool did not alter AChE activity. Anesthesia with thymol was reversed by exposure to picrotoxin (GABAA antagonist), similar to the positive control propofol, but was not reversed by flumazenil (antagonist of benzodiazepine binding site), as observed for the positive control diazepam. Picrotoxin did not reverse the effect of S-(+)-linalool. Thymol exposure at 50 mg/L is more suitable than carvacrol for anesthesia in silver catfish, because this concentration did not cause any mortality or interference with AChE activity. Thymol interacted with GABAA receptors, but not with the GABAA/benzodiazepine site. In contrast, S-(+)-linalool did not act in GABAA receptors in silver catfish.

[Changes in phytochemical and morpho-physilogical traits of thyme [Thymus vulgaris CV varico 3] under different salinity levels]

Background: Thyme is a valuable medicinal plants that evaluation of its quality and quantity yield is necessary

Objective: This study aimed to investigate the effect of salinity on morphological, physiological and phytochemical traits of thyme

Methods: The study was carried out as a randomized complete block design at the Institute of Medicinal Plants with 4 levels of salinity treatments including [control, 50, 100 and 150 mM] and three replications. In this study some morphophysiologial and phytochemical traits of thyme were measured

Results: Results showed the significant effect of salinity on plant height, number of lateral shoots, vegetative dry weight, root fresh weight, root dry weight, Length and Width of a Leaf. The results of the means comparison showed that by increasing levels of salinity, sodium content, electrolyte leakage and proline content increased conversely, the amount of potassium, relative water content, chlorophyll and carotenoid were decreased. The content of thymol and carvacrol increased by higher levels of salinity [100 mM sodium chloride] compared with control

Conclusion: Although the majority of yield related traits negatively affected by salinity but the highest amount of thymol and carvacrol has been achieved in 100 mM NaCl

[Expression of genes 1-deoxy-d-xyhilose 5-phosphate reductoisomerase [dxr], sabinen synthase [tctpsl] and a- terpineol synthase [tctpss] and its relation to thymol and carvaerol biosynthesis in thymus vulgaris under water deficit stress]

Background: Thymol and Carvaerol are the two important secondary metabolites from Thymus vulgaris that productions of them are controlled by genetic and environmental factors

Objective: Evaluation of Water stress effects on expression of three important genes of thymol and carvacrol biosynthetic pathway and also physiological and phytochemical properties of Thymus vulgaris

Methods: The present study was conducted in a randomized complete block design with 4 treatments and 3 replications in Institute of Medicinal Plants, ACECR, Karaj

For this purpose, the treatments were selected as control [FC], 70% of FC, 40% of FC, and 20% of FC. Gene expression was studied using real-time PCR method, and HPLC was applied to essence analysis. Also physiological characteristic including chlorophyll content, relative water content, electrolyte leakage percentage and carotenoids content were analyzed

Results: The results revealed that water stress significantly influenced the gene expression [P< 0.05]. The highest gene expression of DXR and TctpsS was observed in 70% of FC, while the highest one of TvTPSl was recorded in 40% of FC. Moreover, the maximum thymol and carvacrol was found in 70% of FC. In other hand the highest chlorophyll content and corotenoids content were obtained in 100% and 70% of FC respectively. Also there were not significant differences between treatments for relative water content and electrolyte leakage percentage

Conclusion: The 70% of FC through influencing of genes in the firs and last of MEP pathway increased thymol and carvacrol production

Atividade antisséptica do óleo essencial de Lippia origanoides Cham. (Alecrim-pimenta) na presença de leite bovino / Antiseptic activity of the essential oil of Lippia origanoides in the presence of bovine milk

Objetivou-se determinar atividade antisséptica do óleo essencial de Lippia origanoides na presença de leite bovino. A composição química do óleo essencial de alecrim pimenta foi determinada por cromatografia gasosa acoplada à espectrometria de massas (CG-EM), sendo detectados 53 compostos, dos quais 16 foram identificados (>0,1% área total). O carvacrol (32,7%), p-cimeno (23%), timilmetil éter, cariofileno (7,98%) e o γ-terpineno (5,40%) foram os componentes mais abundantes. A concentração inibitória mínima para Staphylococcus aureus ATCC 25923 e E. coli ATCC 8739 foi de 60µL/mL, enquanto para Salmonella Choleraesuis ATCC 35640 foi de 90µL/mL. A concentração bactericida mínima foi de 120µL/mL para as três bactérias. Avaliou-se a atividade antisséptica do óleo essencial na concentração de 120µL/mL na presença de leite bovino sobre as mesmas cepas bacterianas. O óleo em estudo apresentou efeito inibitório do crescimento das cepas em diferentes tempos de ação (p<0,05). S. aureus apresentou maiores índices de inibição após 5 min de contato e Escherichia coli e Salmonella Choleraesuis após 15 min. O óleo essencial de alecrim-pimenta revelou ser um potencial antimicrobiano natural, mesmo na presença de matéria orgânica constituída de uma matriz nutricional complexa que é o leite bovino.(AU)

This study aimed to determine the antiseptic activity of the essential oil of Lippia origanoides in the presence of bovine milk. The essential oil chemical composition was determined by gas chromatography-mass spectrometry (CG EM) 53 compounds were detected, of which 16 were identified (>0.1% total area). The carvacrol (32.7%), p-cymene (23%), thymil methyl ether (10.03%), caryophyllene (7.98 %) and γ-terpinene (5.40%) were the most abundant components. The minimum inhibitory concentration for Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 8739 was 60uL/mL, as for this Salmonella Choleraesuis ATCC 35640 was 90uL/mL. The minimum bactericidal concentration was 120µL/mL for all three bacteria. We evaluated the antiseptic activity of the essential oil in the concentration of 120µL/mL in the presence of bovine milk for the same bacterial strains. The oil under study It showed an inhibitory effect of growth of the strains in different action times (p <0.05). S. aureus showed higher inhibition rates after 5 min of contact and E. coli and Salmonella Choleraesuis after 15 min. The essential oil Lippia origanoides proved to be a natural antimicrobial potential even in the presence of organic matter consists of a complex nutritional matrix is bovine milk.(AU)

Assessment of the effects of decontamination and storage methods on the structural integrity of human enamel / Avaliação dos efeitos de métodos de descontaminação e armazenamento sobre a integridade estrutural do esmalte humano

Introduction: The storage of teeth for use in research is a controversial issue with no consensus on the most appropriate treatment method for this purpose. Objective: The aim of this study was analyze the effectiveness and the influence of different methods of decontamination and storage of human enamel samples, in order to maintain their integrity. Material and method: The sample consisted of 124 molars distributed randomly into three groups according to the method: control - distilled water, 0.1% thymol and 0.02% sodium azide. The tests performed were laser fluorescence, surface microhardness and profilometry analysis (0, 15 and 30 days) and Microbiological test (7, 15 and 30 days). Data were analyzed by the ANOVA and Tukey tests (P <0.05). Result: In the laser fluorescence test, thymol proved to be more feasible for maintaining the integrity of the dental organ, since it did not show significant variations in values among the analyzed periods (P> 0.05). The surface microhardness analysis showed loss of tooth structure in all methods, and sodium azide led to a lower level of tooth loss. Profilometry analysis showed loss of mass in all groups whereas sodium azide showed the greatest loss. None of the methods was able to inhibit bacterial growth. Conclusion: Among the processing methods analyzed none was able to combine effective decontamination and storage with maintenance of the structural integrity of the human enamel.

Introdução: O armazenamento de dentes para utilização em pesquisas é uma questão controvérsia e não há consenso sobre o método de tratamento mais apropriado para esta finalidade. Objetivo: O objetivo deste estudo foi analisar a efetividade e a influência de diferentes métodos de descontaminação e armazenamento do esmalte dentário humano, a fim de manter sua integridade. Material e método: A amostra foi constituída por 124 molares distribuídos aleatoriamente em três grupos, de acordo com o método: controle – água destilada, soluções de timol a 0,1% e de azida de sódio a 0,02%. Os testes realizados foram fluorescência a laser, microdureza de superfície e análise de perfilometria (0, 15 e 30 dias) e teste microbiológico (7, 15 e 30 dias). Os dados foram analisados por meio dos testes ANOVA e Tukey (p<0,05). Resultado: No teste de fluorescência a laser, a solução de timol a 0,1% demonstrou ser mais viável na manutenção da integridade do órgão dentário, uma vez que este não apresentou variações significativas nos valores entre os períodos avaliados (p<0,05). A análise de microdureza de superfície evidenciou perda de estrutura dentária em todos os métodos, sendo que a azida de sódio promoveu menor perda dentária. Na análise de perfilometria observou-se perda de estrutura em todos os grupos, com maior perda no grupo azida de sódio. Nenhum dos métodos conseguiu inibir o crescimento bacteriano. Conclusão: Dentre os métodos de processamento analisados nenhum foi capaz de aliar a efetividade na descontaminação ao armazenamento com manutenção da integridade estrutural do esmalte dentário humano.

Caracterização farmacêutica de nanocápsulas de timol e avaliação da permeação cutânea e da atividade repelente contra Aedes aegypti

A dengue é uma doença infecciosa que pode se manifestar de forma grave, com quadros hemorrágicos. O agente etiológico é um arbovirus transmitido pela picada do mosquito Aedes aegypti, principal vetor da doença. As principais medidas de combate são controle do mosquito e proteção individual, que pode dar-se com o uso de repelentes. Quase a totalidade dos repelentes no mercado possui DEET como ativo, que possui restrições de uso. Nestes entido, o timol (monoterpeno) tem-se mostrado promissor como repelente de inseto (Ae.aegypti), sendo a agregação de tecnologias essencial para a viabilidade de um produto repelente à base de timol. Diante do exposto, o objetivo do presente trabalho foi a preparação e caracterização farmacêutica de nanocápsulas de timol, com avaliação da permeação cutânea, citotoxicidade e atividade repelente de inseto (Ae. aegypti). Para tanto, foi desenvolvido evalidado método analítico para identificação e quantificação do timol por CLAE-DAD nas nanocápsulas de timol (NCT – 1%), que apresentou um teor de ativo em torno de 85%.Prosseguindo a caracterização das NCT, foram determinados o diâmetro médio, índice depolidispersão (PDI), potencial zeta (PZ), eficiência de encapsulação e pH, além do estudo de estabilidade. Análises das NCT mostraram um diâmetro médio de aproximadamente 150 nm,PZ negativo (-27,83 ± 2,60), PDI abaixo de 0,2 e uma eficiência de encapsulação de 98 %...

Dengue is an infectious disease that may also happen in a severe form with hemorrhagicevents. The etiological agent of the disease is an arbovirus wich is transmitted by themosquito Aedes aegypti, the primary vector of the disease. The main actions to combat thedisease are mosquito control and personal protection that can take place using repellents.Almost all repellents have DEET as active substance, which has use restrictions. In thiscontext, thymol (monoterpene) has become a potential insect repellent against Ae. aegypti,and the use of technologies is essential to the viability of a repellent thymol based product.Given the above, the aim of this study was the preparation and pharmaceuticalcharacterization of thymol nanocapsules and evaluation of skin permeation, in vitrocytotoxicity and mosquito repellence (Ae. aegypti). For this purpose, we developed andvalidated analytical method for identification and quantification of thymol by HPLC-PDA.Nanocapsules thymol (NCT - 1%) showed an active content of 85%. Continuing thecharacterization, NCT were analyzed for the mean diameter (D), polydispersity index (PDI),potential zeta (PZ), encapsulation efficiency, pH and product stability . Results shared showedan average diameter of approximately 150 nm, negative PZ (-27,83 ± 2,60), PDI less than 0.2and an encapsulation efficiency of 98%...