Basil, tea tree and clove essential oils as analgesics and anaesthetics in Amphiprion clarkii (Bennett, 1830) / Óleos essenciais de manjericão, melaleuca e cravo como anestésicos e analgésicos em Amphiprion clarkii (Bennett, 1830)

Abstract In this study were evaluated the anaesthesia and analgesic effects of clove Eugenia caryophyllata, tea tree Melaleuca alternifolia and basil Ocimum basilicum essential oils (EO) during handling of yellowtail clownfish Amphiprion clarkii. Juveniles (3.70 ± 0.75 cm and 1.03 ± 0.50 g; mean ± standard deviation) were submitted to concentrations of 40, 50, 60, 70 and 80 µl L-1 of clove, 150, 200, 250, 300 and 350 µl L-1 of basil and 200, 300, 400, 500 and 600 µl L-1 of tea tree oils (n=10/concentration), previously defined in pilot tests. Individually and only once, fish from each treatment were placed in a glass recipient containing 1 L of seawater at a temperature of 25 °C, salinity of 35 g L-1 and the specific concentration of diluted EO (stock solution). Control (only seawater) and blank (seawater and ethanol at the highest concentration used to dilute the oils) treatments were also conducted. After reaching the stage of surgical anaesthesia, fish were submitted to biometry and a sensibility test. After that, they were transferred to clean seawater for anaesthesia recovery. The times of induction needed to reach each anaesthesia stage and anaesthesia recovery were recorded. Animals were observed for 72 hours after the procedures. All the EO provoked anaesthesia and analgesic effects in A. clarkii, but basil oil is not recommended because it caused involuntary muscle contractions and mortality in 100% and 12% of fish, respectively. The lower concentrations that promote suitable induction and recovery times are 50 µl L-1 of clove oil and 500 µl L-1 of tea tree oil. However, due to its complementary high analgesic efficiency, clove oil is recommended as the ideal anaesthetic for A. clarkii.

Resumo Neste estudo foram avaliados os efeitos anestésicos e analgésicos dos óleos essenciais (OE) de cravo Eugenia caryophyllata, melaleuca Melaleuca alternifolia e manjericão Ocimum basilicum durante manejo de peixes-palhaços Amphiprion clarkii. Juvenis (3.70 ± 0.75 cm e 1.03 ± 0.50 g; média ± desvio padrão) foram submetidos às concentrações de 40, 50, 60, 70 e 80 µl L-1 de cravo, 150, 200, 250, 300 e 350 µl L-1 de manjericão e 200, 300, 400, 500 e 600 µl L-1 de melaleuca (n=10/concentração), previamente definidas em testes pilotos. Individualmente e somente uma vez, os peixes de cada tratamento foram colocados em recipiente de vidro contendo 1 L de água salgada, em temperatura de 25 °C, salinidade de 35 g L-1 e a concentração específica de OE diluída (solução estoque). Tratamentos controle (apenas água marinha) e branco (água marinha e a maior concentração de etanol utilizada para diluição dos óleos) também foram conduzidos. Após atingirem o estágio de anestesia cirúrgica, os peixes foram submetidos à biometria e teste de sensibilidade. Em seguida, foram transferidos para água marinha limpa. Os tempos necessários para atingir cada estágio anestésico e recuperação foram registrados. Os animais foram observados por 72 horas após os procedimentos. Todos os OE provocaram anestesia e analgesia em A. clarkii, porém o óleo de manjericão não é recomendado, pois causou contrações musculares involuntárias e mortalidade em 100% e 12% dos animais, respectivamente. As menores concentrações que promovem indução anestésica e recuperação em tempos adequados são 50 µl L-1 de óleo de cravo e 500 µl L-1 de óleo de melaleuca. Entretanto, devido à sua alta eficiência analgésica complementar, o óleo de cravo é recomendado como o anestésico ideal para A. clarkii.

Basil, tea tree and clove essential oils as analgesics and anaesthetics in Amphiprion clarkii (Bennett, 1830)

Abstract In this study were evaluated the anaesthesia and analgesic effects of clove Eugenia caryophyllata, tea tree Melaleuca alternifolia and basil Ocimum basilicum essential oils (EO) during handling of yellowtail clownfish Amphiprion clarkii. Juveniles (3.70 ± 0.75 cm and 1.03 ± 0.50 g; mean ± standard deviation) were submitted to concentrations of 40, 50, 60, 70 and 80 µl L-1 of clove, 150, 200, 250, 300 and 350 µl L-1 of basil and 200, 300, 400, 500 and 600 µl L-1 of tea tree oils (n=10/concentration), previously defined in pilot tests. Individually and only once, fish from each treatment were placed in a glass recipient containing 1 L of seawater at a temperature of 25 °C, salinity of 35 g L-1 and the specific concentration of diluted EO (stock solution). Control (only seawater) and blank (seawater and ethanol at the highest concentration used to dilute the oils) treatments were also conducted. After reaching the stage of surgical anaesthesia, fish were submitted to biometry and a sensibility test. After that, they were transferred to clean seawater for anaesthesia recovery. The times of induction needed to reach each anaesthesia stage and anaesthesia recovery were recorded. Animals were observed for 72 hours after the procedures. All the EO provoked anaesthesia and analgesic effects in A. clarkii, but basil oil is not recommended because it caused involuntary muscle contractions and mortality in 100% and 12% of fish, respectively. The lower concentrations that promote suitable induction and recovery times are 50 µl L-1 of clove oil and 500 µl L-1 of tea tree oil. However, due to its complementary high analgesic efficiency, clove oil is recommended as the ideal anaesthetic for A. clarkii.

Resumo Neste estudo foram avaliados os efeitos anestésicos e analgésicos dos óleos essenciais (OE) de cravo Eugenia caryophyllata, melaleuca Melaleuca alternifolia e manjericão Ocimum basilicum durante manejo de peixes-palhaços Amphiprion clarkii. Juvenis (3.70 ± 0.75 cm e 1.03 ± 0.50 g; média ± desvio padrão) foram submetidos às concentrações de 40, 50, 60, 70 e 80 µl L-1 de cravo, 150, 200, 250, 300 e 350 µl L-1 de manjericão e 200, 300, 400, 500 e 600 µl L-1 de melaleuca (n=10/concentração), previamente definidas em testes pilotos. Individualmente e somente uma vez, os peixes de cada tratamento foram colocados em recipiente de vidro contendo 1 L de água salgada, em temperatura de 25 °C, salinidade de 35 g L-1 e a concentração específica de OE diluída (solução estoque). Tratamentos controle (apenas água marinha) e branco (água marinha e a maior concentração de etanol utilizada para diluição dos óleos) também foram conduzidos. Após atingirem o estágio de anestesia cirúrgica, os peixes foram submetidos à biometria e teste de sensibilidade. Em seguida, foram transferidos para água marinha limpa. Os tempos necessários para atingir cada estágio anestésico e recuperação foram registrados. Os animais foram observados por 72 horas após os procedimentos. Todos os OE provocaram anestesia e analgesia em A. clarkii, porém o óleo de manjericão não é recomendado, pois causou contrações musculares involuntárias e mortalidade em 100% e 12% dos animais, respectivamente. As menores concentrações que promovem indução anestésica e recuperação em tempos adequados são 50 µl L-1 de óleo de cravo e 500 µl L-1 de óleo de melaleuca. Entretanto, devido à sua alta eficiência analgésica complementar, o óleo de cravo é recomendado como o anestésico ideal para A. clarkii.

In vitro Antimicrobial Activity of the Flower Buds of Eugenia caryophyllata.

Aims: This study investigated the antimicrobial effect of aqueous and ethanol extracts of the flower buds of Eugenia caryophyllata (Myrtaeae) against a wide range of bacteria and yeasts cells isolated clinically from patients. Methodology: The agar diffusion method was used to establish the antimicrobial activity and the zones of inhibition caused by the extracts. The antimicrobial effects of 16% and 32% aqueous and ethanol extracts of Eugenia caryophyllata were investigated against 111 pathogenic bacteria and yeasts cells. The microbes used consisted of 11 Proteus mirabilis, 20 Salmonella typhi, 15 Pseudomonas aeruginosa, 18 Escherichia coli, 19 Staphylococcus aureus, 12 Klebsiella pneumoniae, and 16 Candida albicans species. Results: The ethanol extracts inhibited the growth of all the microbes employed in the study with inhibition zones ranging from 8.00±0.00 mm to 24.00±0.00 mm. The aqueous extracts however exhibited different degrees of antimicrobial activity with zones of inhibition ranging from 6.00±0.00 mm to 13.33±0.29 mm. Conclusion: Our study concludes that the aqueous and ethanol extracts of the flower buds of Eugenia caryophyllata have relatively good antimicrobial activity against a wide range of medically important pathogenic bacteria and Candida albicans in vitro.

In vitro assessment of antimicrobial activity of the selected essential oils.

The present study evaluates the antimicrobial activity of three essential oils of Eucalyptus globulus, Eugenia caryophyllata and Psoralea corylifolia. The agar well diffusion method was used to assess the antimicrobial activity against five selected microorganisms S. aureus and B. subtilis (Gram-positive bacteria); E. coli and P. aeruginosa (Gram-negative bacteria) and P. chrysogenum (Fungus). The results revealed that the E. globulus was found to be most significant in comparison to the other oils. All the three oils have revealed higher antimicrobial activity against S. aureus.

Ensayo preliminar de la actividad antibacteriana de extractos de Allium sativum, Coriandrum sativum, Eugenia Caryophyllata, Origanum vulgare, Rosmarinus officinalis Y Thymus vulgaris frente a Clostridium perfringens / Preliminary test of the antibacterial activity of the Allium sativum, Coriandrum sativum, Eugenia

Se evaluó la actividad antibacteriana frente a Clostridium perfringens (cepa ATCC: 13124) por el método de Kirby Bauer en agar SPS de los aceites esenciales o extractos vegetales obtenidos con solventes orgánicos de diferente polaridad a partir de Allium sativum (ajo), Coriandrum sativum (cilantro), Eugenia Caryophyllata (clavode olor), Origanum vulgare (orégano), Rosmarinus officinalis (romero) y Thymus vulgaris (tomillo), utilizando la vancomicina como control. Los extractos obtenidos por el método de lixiviación de O. vulgare y T. vulgaris no presentaron inhibición para este microorganismo; los demás extractos vegetales sí la presentaron, obteniéndose concentraciones bacteriostáticas mínimas que oscilaron entre 16 y 63 μl/ml. El extracto etanólico y el aceite esencial de E.caryophyllata fueron los que presentaron una menor concentración inhibitoria mínima (250 μl/ml). Se observan variaciones importantes enla capacidad de inhibición de dichos extractos con respecto a estudios realizados por otros grupos de investigación en el mundo, pocos de ellos utilizaron a Clostridium perfringens.

The antibacterial activity against Clostridium perfringens (ATCC: 13124) of essential oils and extracts obtained from Allium sativum (garlic), Coriandrum sativum (coriander), Eugenia Caryophyllata (clove), Origanum vulgare (oregano), Rosmarinus officinalis (rosemary) and Thymus vulgaris (thyme) tested by means of the Kirby Bauer method in SPS agar, using vancomicine as the control. The extracts obtained by the lixiviation method from Origanum vulgare and Thymus vulgaris do not show inhibition, whereas the other extracts had minimal bacteriostatical concentrations between 16 to 63 ìl/ml. The E. caryophyllata ethanolic extract and essential oil presented a lower minimal inhibitory concentration (250 ìl/ml). Important variations in the inhibitory capacity of the extracts and essential oils were observed, regarding studies carried out by other research groups in the world, few worked with Clostridium perfringes.

Evaluación del efecto antioxidante de aceites esenciales y extractos de Eugenia caryophyllata, Origanum vulgare Y Thymus vulgaris / Evaluation of antioxidant effect of essential oils and extracts of eugenia caryophyllata, origanum

A partir de aceites esenciales y extractos de Eugenia Caryophyllata, Origanum vulgare y Thymus vulgaris, y después de analizar su potencialantioxidante, se seleccionaron los que mayor inhibición a la oxidación presentaron, para ser analizados en diferentes concentraciones y encomparación con un antioxidante referencia (nitrito sódico) sobre la oxidación de lípidos y su efecto sobre las características organolépticas en productos cárnicos cocidos como el salami. La actividad antioxidante de los diferentes aceites esenciales se evalúo por varios métodos (grado de inhibición de la peroxidación del ácido linoleico, evaluación del poder de reducción, test del β-caroteno), con el fin de demostrar que los tratamientos con aceites esenciales de orégano, tomillo y clavo de olor reducen significativamente la oxidación en productos cárnicos cocidos como el salami; estos fueron evaluados de acuerdo a los resultados de actividad antioxidante, antibacterial y concentración mínima inhibitoria (MIC).Obteniéndose resultados positivos para el clavo de olor, el cual fue seleccionado para adicionar a la formulación del salami, como antioxidante y como conservante. La concentración de aplicación en el salami se realizó con el extracto de clavo de olor. La elaboración del salami, se efectuó con base en los procedimientos establecidos en el manual de prácticas y formulaciones de la Unidad Tecnológica de Alimentos de la Universidad deCaldas. Al producto se le realizó una serie de análisis sensoriales, para determinar el efecto del extracto adicionado como antioxidante sobre las propiedades organolépticas del salami;los cuales arrojaron resultados positivos. El producto no mostró grandes diferencias, con respecto al patrón, siendo aceptado en las pruebas sensoriales, por los jueces.

After analyzing their antioxidant potential, the essential oils and extracts of Eugenia caryophyllata, Origanum vulgare and Thymus vulgaris with the greatest inhibition to oxidation were selected in order to be analyzed at different concentrations and compared with a reference antioxidant (sodium nitrite) on lipid oxidation and their effect on the organoleptics characteristics in cooked meat products like salami. The antioxidant activity of different essential oils was evaluated by means of several methods (peroxidation inhibition degree of linoleic acid, reduction power assessment, ß-carotene test), in order to demonstrate that treatments with oregano, thyme and clove essential oils significantly reduce oxidation in cooked meat products like salami. These oils were evaluated according to the results of antioxidant and antibacterial activity and the minimum inhibitory concentration (MIC). Positive results were also obtained for clove, which was selected to be added to the formulation of salami, as an antioxidant and preservative. The concentration of the application in salami was carried out with the clove extract. The salami was elaborated based on the procedures established in the practices and formulations manual of the Food Technology Unit of the Universidad de Caldas. The product underwent a series of sensory analysis to determine the effect of the extract added as an antioxidant on the organoleptic properties of salami, which showed positive results. The product showed no significant differences with regards to the pattern, accepted by the judges in the sensory assessment.

Anti-inflammatory and antinociceptive activities a of eugenol essential oil in experimental animal models / Atividades antiinflamatória e antinociceptiva do eugenol em modelos experimentais em animais

Eugenia caryophyllata, popularmente conhecida como "cravo-da-índia", cresce naturalmente na Indonésia e é cultivada em várias partes do mundo, incluindo o Brasil. O cravo-da-índia é utilizado em culinária, em farmácia, perfumaria e cosméticos. O óleo essencial extraído do cravo-da-índia cujo principal componente é o eugenol tem sido utilizado em odontologia como anti-séptico e analgésico. O objetivo deste estudo foi avaliar as atividades antiinflamatória e antinociceptiva do eugenol de uso odontológico, administrado oralmente, em modelos experimentais in vivo. A atividade antiinflamatória do eugenol foi avaliada através do volume de exsudato e migração leucocitária no teste de pleurisia e do edema de pata de rato induzido pela carragenina. A atividade antinociceptiva foi avaliada através dos testes de contorções induzidas pelo ácido acético e da placa quente. O eugenol (200 e 400 mg/kg) reduziu o volume de exsudato pleural sem interferir na contagem de leucócitos totais presentes na pleura. Na dose de 200 mg/kg, o eugenol inibiu significativamente o edema de pata, 2-4 h após a injeção do agente flogístico. No teste da placa quente, a administração do eugenol (100 mg/kg) mostrou atividade significativa à reação de desconforto-tempo dependente, avaliada como a latência da resposta, inibida pela meperidina. Eugenol na doses de 50, 75 e 100 mg/kg apresentou efeito antinociceptivo significativo no teste de contorções abdominais induzidas pelo ácido acético em comparação com o grupo controle. Os dados obtidos indicam que o eugenol apresenta atividade antiinflamatória e antinociceptiva periférica.

Eugenia caryophyllata, popular name "clove", is grown naturally in Indonesia and cultivated in many parts of the world, including Brazil. Clove is used in cooking, food processing, pharmacy; perfumery, cosmetics and the clove oil (eugenol) have been used in folk medicine for manifold conditions include use in dental care, as an antiseptic and analgesic. The objective of this study was evaluated the anti-inflammatory and antinociceptive activity of eugenol used for dentistry purposes following oral administration in animal models in vivo. The anti-inflammatory activity of eugenol was evaluated by inflammatory exudates volume and leukocytes migration in carrageenan-induced pleurisy and carrageenan-induced paw edema tests in rats. The antinociceptive activity was evaluated using the acetic acid-induced writhing and hot-plate tests in mice. Eugenol (200 and 400 mg/kg) reduced the volume of pleural exudates without changing the total blood leukocyte counts. At dose of 200 mg/kg, eugenol significantly inhibited carrageenan-induced edema, 2-4 h after injection of the flogistic agent. In the hot-plate test, eugenol administration (100 mg/kg) showed unremarkable activity against the time-to-discomfort reaction, recorded as response latency, which is blocked by meperidine. Eugenol at doses of 50, 75 and 100 mg/kg had a significant antinociceptive effect in the test of acetic-acid-induced abdominal writhing, compared to the control animals. The data suggest that eugenol possesses anti-inflammatory and peripheral antinociceptive activities.