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.

Efeito da Astrocaryum aculeatum (Tucumã) na toxicidade da Doxorrubicina: modelo experimental in vivo / Effect of Astrocaryum aculeatum (tucumã) on doxorubicin toxicity: in vivo experimental model

Resumo Objetivo Obter o óleo do Astrocaryum aculeatum (A.a) e avaliar a genotoxidade/antigenotoxidade pelo teste do micronúcleo em células do sangue periférico. Métodos O óleo da A.a foi obtido por prensagem hidráulica. Os animais foram camundongos Swiss, machos e saudáveis com 6-7 semanas de idade, 6 por grupo. Teste genotóxico e antigenotóxico as concentrações foram de 500, 1.000 e 2.000 mg/kg por 0,5 mL (via oral), seguidas ou não de injeção intraperitoneal de doxorrubicina (0,3mL - 15 mg/kg por peso corporal), além do grupo negativo (água) e dimetilsufóxido (600 µL). As amostras de sangue periférico foram coletadas 24h e 48h após o tratamento. Resultados Houve redução estatisticamente significativa na frequência de micronúcleos em células policromáticas que variou de 34,72% à 38,19% para os tratamentos de 24h, e de 63,70 à 66,12% para os de 48h. Conclusão O óleo fixo do tucumã apresentou potencial antigenotóxico para as concentrações em tratamentos agudos.

Abstract Objective To obtain the oil of Astrocaryum aculeatum (A.a), and evaluate its genotoxicity/antigenotoxicity activities using the micronucleus test in peripheral blood cells. Methods The oil of Astrocaryum aculeatum was obtained by hydraulic pressing. The animals used were healthy Swiss male mice, at 6-7 weeks of age; there were six per group. The genotoxic and antigenotoxic activity of concentrations were 500, 1,000 and 2,000 mg/kg per 0.5 mL (oral), followed or not followed by intraperitoneal injection of doxorubicin (0.3 mL-15 mg/kg by body weight), in addition to a negative group (water) and dimethyl sulfoxide (600 μL). Peripheral blood samples were collected 24h and 48h after treatment. Results A statistically significant reduction was identified in the frequency of micronuclei in polychromatic cells ranging from 34.72% to 38.19% for 24-hour treatments, and from 63.70% to 66.12% for 48 hour. Conclusion The fixed oil of tucumã presented antigenotoxic potential for the concentrations used in acute treatments.

Evaluación de la toxicidad del aceite esencial de Aloysia triphylla Britton (cedrón) y de la actividad anti-Trypanosoma cruzi del citral, in vivo / In vivo evaluation of Aloysia triphylla britton (lemon verbena) essential oil toxicity and citral anti-Trypanosma cruzi activity

Existe escasa investigación en enfermedades olvidadas. Las plantas medicinales son una potencial fuente de compuestos antimicrobianos. Objetivos: Determinar la toxicidad del aceite esencial de Aloysia triphylla y la actividad del citral contra Trypanosoma cruzi en ratones. Diseño: Estudio experimental preclínico in vivo. Institución: Facultad de Medicina, Universidad Nacional Mayor de San Marcos, Lima, Perú. Material: Ratones albinos. Intervenciones: La toxicidad aguda oral a dosis única fue evaluada en ratas albinas. Para la actividad tripanocida se utilizaron ratones asignados a los siguientes grupos: infectados y no tratados (G1), infectados y tratados con citral en dosis de 50, 150 y 300 mg/kg/día (G2, G3 y G4, respectivamente), infectados y tratados con benznidazol 100 mg/kg (G5) y no infectados y no tratados (G6). La parasitemia fue determinada individualmente cada 2 días por microscopia directa. En los días 14, 21 y 28 post infección, cinco ratones de cada grupo fueron sacrificados y los corazones procesados para análisis histopatológico. Principales medidas de resultados: Signos de toxicidad y mortalidad, y parasitemia. Resultados: La dosis límite de 2 000 mg/kg no provocó signos ni síntomas de toxicidad y los estudios anatomopatológicos macroscópicos y microscópicos no mostraron alteración de los órganos estudiados. La parasitemia fue reducida significativamente con la dosis de 300 mg/kg en los días 16, 18, y 20 post infección (p<0,05). El número de nidos de amastigotes y de infiltrados inflamatorios en corazón fueron reducidos en 67,7 por ciento y 51,7 por ciento, respectivamente, con 300 mg/kg en el día 28. Conclusiones: El aceite esencial de Aloysia triphylla es calificado como no tóxico y el citral en dosis de 300 mg/kg tuvo actividad contra Trypanosoma cruzi en ratones...

There is limited research on neglected diseases. Medicinal plants are potential sources of antimicrobial compounds. Objectives: To determine the toxicity of Aloysia triphylla essential oil and citral activity against Trypanosoma cruzi in mice. Design: Experimental study in vivo, preclinical. Setting: Faculty of Medicine, Universidad Nacional Mayor de San Marcos, Lima, Peru. Biological material: Albino mice. Main outcome measures: Signs of toxicity and mortality and parasitemia. Interventions: Acute oral toxicity at single dose was evaluated in albino rats. For trypanocidal activity mice were assigned to the following groups: untreated infected (G1), infected and treated with citral at doses 50, 150 and 300 mg/kg/day (G2, G3 and G4 respectively), infected and treated with benznidazole 100 mg/kg (G5), and uninfected and untreated (G6). Parasitemia was determined individually every 2 days by direct microscopy. In days 14, 21 and 28 post infection five mice from each group were sacrificed and their hearts processed for histopathology. Results: The limit dose of 2 000 mg/kg did not cause signs or symptoms of toxicity and macro and microscopic anatomopathology did not show alterations in the organs studied. Parasitemia was significantly reduced at dose of 300 mg/kg at days 16, 18, and 20 post infection (p<0.05); the number of amastigote nests and inflammatory infiltrates in heart were reduced on day 28 by 67.7 per cent and 51.7 per cent respectively with 300 mg/kg. Conclusions: Aloysia triphylla essential oil is qualified as nontoxic and citral at 300 mg/kg dose had activity against Trypanosoma cruzi in mice...

Carum copticum and Thymus vulgaris oils inhibit virulence in Trichophyton rubrum and Aspergillus spp

Emergence of drug-resistant strains has demanded for alternative means of combating fungal infections. Oils of Carum copticum and Thymus vulgaris have long been used in ethnomedicine for ailments of various fungal infections. Since their activity has not been reported in particular against drug-resistant fungi, this study was aimed to evaluate the effects of oils of C. copticum and T. vulgaris on the growth and virulence of drug-resistant strains of Aspergillus spp. and Trichophyton rubrum. The gas chromatography-mass spectrometry analysis revealed thymol constituting 44.71% and 22.82% of T. vulgaris and C. copticum, respectively. Inhibition of mycelial growth by essential oils was recorded in the order of thymol > T. vulgaris > C. copticum against the tested strains. RBC lysis assay showed no tested oils to be toxic even up to concentration two folds higher than their respective MFCs. Thymol exhibited highest synergy in combination with fluconazole against Aspergillus fumigatus MTCC2550 (FICI value 0.187) and T. rubrum IOA9 (0.156) as determined by checkerboard method. Thymol and T. vulgaris essential oil were equally effective against both the macro and arthroconidia growth (MIC 72 µg/mL). A > 80% reduction in elastase activity was recorded for A. fumigatus MTCC2550 by C. copticum, T. vulgaris oils and thymol. The effectiveness of these oils against arthroconidia and synergistic interaction of thymol and T. vulgaris with fluconazole can be exploited to potentiate the antifungal effects of fluconazole against drug-resistant strains of T. rubrum and Aspergillus spp.

possible toxic effect of different doses of Nigella sativa oil on the histological structure of the liver and renal cortex of adult male albino rats

Nigella sativa seeds are commonly known as black seed or black cumin. It has been used for thousands of years as a spice and food preservative and also as a protective and curative remedy for numerous disorders. Our research aimed to study the possible toxic effect of different doses of N. sativa oil on the liver and kidney. Twenty-one adult male albino rats were used and divided equally into three groups. The first group was the control group. The second and third groups received the oil in two gradually increasing doses of 15 and 25 ml/kg, respectively, for 1 month. The animals were then sacrificed and samples of liver and kidney were taken and prepared for histological examination. In the kidney of group II there was epithelial shedding and necrosis of some cells of the proximal and distal convoluted tubules, but there was no effect on renal glomeruli. In contrast, in group III there was glomerular injury in the form of degeneration of the tuft of capillaries, ill-defined basement membrane, and destruction of endothelial cells, in addition to tubular necrosis. In group II there was minimal effect on the liver in the form of perivascular cellular infiltration; in group III was seen a markedly vacuolated foamy cytoplasm of hepatocytes, with dilated sinusoids and perivascular cellular infiltration. In conclusion, large doses of N. sativa oil have toxic effects on the histological structure of the kidney and to a lesser degree on the liver. Therefore, Nigella oil should be used in proper doses, and further studies on the effect of large doses of oil are recommended

Impact of galectin-3 deficiency on gene expression in the liver of mice 48-h post-treatment with olive oil or CC1[4]

Galectin-3 [Gal-3] is a multifunctional protein, playing a key role in many biolegical processes. Previous study demonstrated that normal hepatocytes do not express galectin-3, but this protein can be present in injured liver. The present paper aimed to assist in elucidate the biological role of galectin-3 in injured liver by CC1[4] and to clarify genes that differentially expressed in response to galectin-3 deficiency in normal and chemically injured liver of mice 48-h post-treatment with olive oil or CC1[4]. Four male wild type mice [WT] and another four galectin-3 disrupted mice [Gal-3[-/-]] were used in this experiment. The mice were fasted overnight and classified into two groups, [each group including, two WT and two Gal-3[-/-] mice] the first subgroup received in the following morning 4 ml/kg olive oil, while the second subgroup was received 8 ml/kg CC1[4] [50% in olive oil] by gavages. After 48h, the mice were anesthetized and killed to obtain blood and excise the liver. Gene's expression analysis in the liver tissue was carried out using cDNA microarray technique. The cDNA microarrays analysis revealed that 7 genes have clearly changed their levels of expression, of these 5 genes related to detoxification mechanisms are up-regulated and 2 genes related to tumor cell and amyloid protein have been down-regulated in Gal-3[-/-] mice after 48 h post-treatment with olive oil. The mice treated with CC1[4] reveled that 42 genes have clearly changed their levels of expression, of this 8 genes were up-regulated and 34 genes were down-regulated. Of the up-regulated genes were detoxification, fatty acids and lipid metabolism proteins. On the other hand, the down-regulated genes encoded proteins for xenobiotic metabolism, stress response, transcription factors, lipid metabolism, proteolysis and peptedolysis, RNA, nerve system, and immune responses proteins. This study demonstrated that changes in gene expression profile in galectin-3 deficiency mice 48-h post-treatment with CC1[4], mostly related to down-regulated different genes including, many biological processes, implying the multifunctional of galectin-3 to protect and ameliorated the liver injury induced by CC1[4] in mice

Larvicidal activities of Cinnamomum osmophloeum and matricharia chamomella extracts against the filarial mosquito culexquinquefasciatus [diptera:culicidae] and their effects on its haemogram

The larvicidal activities of the bark essential oils of [Cinnamon] Cinnamomum osmophloeum and the leaf essential oil of [Chamomell] Matricharia chamomella were evaluated Culex quinquefasciatus in laboratory. The toxicity index, LC[50] values were 0.260 and 0.310 mg/L for Cinnamomum osmophloeum and Matricharia chamomella, respectively. Four types of haemocytes: prohaemocyte [Pr], Granulocytes [Gr], Plasmatocytes [Pl] and Oenocytoides [Oe] were determined, the present results show that cinnamon and chamomile oils significantly decreased the number of Pr, Gr and Pl and increased the number of Oe. Furthermore, the effect of tested plant oils on the haemocytees surface area were evaluated

Phytotoxicity of volatile oil from Eucalyptus citriodora against some weedy species.

A study was undertaken to explore the phytotoxicity of volatile essential oil from Eucalyptus citriodora Hook. against some weeds viz. Bidens pilosa, Amaranthus viridis, Rumex nepalensis, and Leucaena leucocephala in order to assess its herbicidal activity. Dose-response studies conducted under laboratory conditions revealed that eucalypt oils (in concentration ranging from 0.0012 to 0.06%) greatly suppress the germination and seedling height of test weeds. At 0.06% eucalypt oil concentration, none of the seed of test weeds germinated. Among the weed species tested, A. viridis was found to be the most sensitive and its germination was completed inhibited even at 0.03%. Not only the germination and seedling growth, even the chlorophyll content and respiratory activity in leaves of emerged seedlings were severely affected. In A. viridis chlorophyll content and respiratory activity were reduced by over 51% and 71%, respectively, even at a very low concentration of 0.06%. These results indicated an adverse effect of eucalypt oils on the photosynthetic and energy metabolism of the test weeds. A strong negative correlation was observed between the observed effect and the concentration of eucalypt oil. Based on the study, it can be concluded that oil from E. citriodora possess strong inhibitory potential against weeds that could be exploited for weed management.

Evaluación de la toxicidad aguda y la acción hipolipemiante del aceite de Plukenetia volúbilis, Sacha Inchi / Evaluation of acute toxicity and hypolipemic action of Plukenetia volúbilis, Sacha Inchi

Se evaluó el efecto tóxico agudo del aceite de Plukenetia volúbilis (Sacha Inchi), determinando la Dosis Letal -50 (DL50) en 42 ratones de la cepa Nish, distribuidos en siete grupos de seis ratones cada uno, a los cuales se les administró, vía oral, dosis crecientes del aceite. Para evaluar el efecto hipolipemiante utilizamos 55 ratas albinas machos de la raza Holtzman, con un peso entre 194 g y 290 g y dos meses de edad. Se utilizó una dieta con 15 por ciento de grasas saturadas para elevar los lípidos séricos durante todo el tiempo del experimento. Utilizamos tres dosis diferentes de aceite de Sacha Inti, administradas durante 14 días consecutivos, comparando sus efectos con un control positivo (Lovastatina) y un control negativo (SF). Se controló el peso de los animales y los valores de colesterol, HDL y triglicéridos sanguíneos en tres tiempos diferentes. El análisis estadístico se realizó mediante el programa STATA 8.2. La DL50 fue de 111,65 mg/kg; hubo una reducción significativa de los valores sanguíneos de triglicéridos y LDL, mas no de colesterol y peso de los animales.

The acute toxic of Plukenetia volúbilis (Sacha Inchi) oil was evaluated. We calculated the Medial Lethal Dose (DL50) on 42 mice of the Nish strain, distributed in seven groups of six mice each, to whom a different and increasing dose was administered by oral route. To determine the hypolipidemic effect, we used 55 male albino rats of the Holtzman breed, whose weight fluctuateed between 194 and 290 g. The animals were fed with 15% of a saturateed fat diet for 14 days to produce hyperlipidemia during the whole period of the experiment. Treatment with Sacha Inchi oil in three different doses during 14 consecutive days was compared to the effects of a positive (lovastatine) and a negative control (physiological serum). The weight, cholesterol, HDL, LDL, VLDL, and triglycerides, were controlled in three different periods of time. Statistics analysis was made using the STATA 8.2 program. The DL50 was detrmined at 111,65 mg/Kg and the level of tryclicerides and LDL were statistically reduced, but not the cholesterol values or the animals' weight.

Absence of genotoxic effects of crude and refined red palm oil on mouse bone marrow cells

Red Palm Oil (RPO), extracted from fruits of Elaeis guineensis, is a complex mixture consisting of over 99 per cent glycerides and about 1 per cent non-glyceride compounds. Its orange-red colour is due to its high content of carotenoid pigments, mainly alpha- and beta-carotene. Based on the fact that palm oil is a rich source of provitamin A, and because it is largely consumed in North and Northeastern Brazil, we evaluated possible clastogenic and cytotoxic activities of this oil on mouse bone marrow cells in vivo, as well as the alpha- and beta-carotene content. The experiments were performed using samples of refined and crude palm oil, of which two different phases, supernatant, sediment, and the mixture of both, were tested. The animals were treated by gavage, at daily doses of 4.5 g/Kg, for five consecutive days, and killed 24 hours after the last treatment, for chromosome preparations. The negative control group was treated with corn oil. There was no statistically significant difference in the frequency of chromosomal aberrations and in mitotic index when the animals which received palm oil were compared with the negative control. The beta-carotene content was higher than that of alpha-carotene, and the supematant phase was the richest source of carotenoids. These findings suggest that RPO has no genotoxic effect.

Toxic effects of garlic extract and garlic oil in rats.

Significant rise in urea and D-aspartate aminotransferase and inhibition of alkaline phosphatase in serum were observed in rats fed garlic extract (2 ml/100 g body wt, intragastrically) for 10 days. The liver showed histological changes. Garlic oil feeding (10 mg/100 g body wt, intragastrically) after 24 hr fasting was found lethal. The cause of death appears to be acute pulmonary oedema. On histological examination, all the organs of the dead rats revealed severe congestion. However, similar feeding of garlic oil was well tolerated by rats in the fed state. Also, 24 hr fasted rats could tolerate this dose of garlic oil, provided they were previously adapted to garlic oil feeding.