Antifungal activity of plant extracts against microsporum canis and candida spp. strains / Actividad antifúngica de extractos de plantas contra cepas de microsporum canis y candida spp

Aiming to find a phytotherapeutic compounds to treat animal fungal infections, plants commonly found in Northeastern Brazil were evaluated in vitro against Microsporum canis and Candida spp. strains isolated from dogs and cats. The leaf ethanol extracts of Momordica charantia, Calotropis procera, Peschiera affinis and Piper tuberculatum and decoction of Mangifera indica were initially evaluated by the agar-well diffusion method. Four extracts induced growth inhibition zones against M. canis: P. tuberculatum (20 mm), M. indica (14 mm), M. charantia (13 mm) and P. affinis (11 mm). None of them were active against Candida spp. Broth microdilution tests were performed for M. canis strains (n=5), to find the minimum inhibitory concentration (MIC) and the minimum fungicidal concentration (MFC). The geometric means for the MIC values were 590, 370, 350, 170 ug/mL, and for the MFC values were 1190, 750, 700, 340 ug/mL for M. charantia, P. affinis, P. tuberculatum and M. indica, respectively. Therefore, extracts from M. charantia, P. affinis, P. tuberculatum and M. indica are good candidates to produce antifungal phytotherapics since these extracts demonstrated good activity against M. canis.

Con el objetivo de encontrar compuestos fitoterapéuticos para tratar las infecciones por hongos de los animales, plantas que se encuentran comúnmente en el noreste de Brasil se evaluaron in vitro frente a cepas de Microsporum canis y Candida spp. aisladas de perros y gatos. Los extractos etanólicos de hojas de Momordica charantia, Calotropis procera, Peschiera affinis y Piper tuberculatum y la decocción de Mangifera índica fueron evaluados inicialmente por el método de difusión en pocillos de agar. Cuatro extractos indujeron zonas de inhibición del crecimiento contra M. canis: P. tuberculatum (20 mm), M. índica (14 mm), M. charantia (13 mm) y P. affinis (11 mm). Ninguno de ellos fue activo contra Candida spp. Se realizaron pruebas de microdilución en caldo para las cepas de M. canis (n = 5), para encontrar la concentración mínima inhibitoria (CIM) y la concentración fungicida mínima (CFM). Las medias geométricas de los valores de CIM fueron 590, 370, 350, 170 mg/ml, y para los valores de CFM fueron 1.190, 750, 700, 340 mg/ml de M. charantia, P. affinis, P. tuberculatum y M. indica, respectivamente. Por lo tanto, los extractos de M. charantia, P. affinis, P. tuberculatum y M. indica son buenos candidatos para la producción de fitoterápicos antifúngicos ya que estos extractos demostraron una buena actividad contra M. canis.

Leishmanicidal and cholinesterase inhibiting activities of phenolic compounds of Dimorphandra gardneriana and Platymiscium floribundum, native plants from Caatinga biome / Atividade inibitória Leishmanicida e Colinesterásica de Compostos Fenólicos de Dimorphandra gardneriana e Platymiscium fliribunbum, plantas nativas do bioma Caatinga

In recent years, the Brazilian Health Ministry and the World Health Organization have supported research into new technologies that may contribute to the surveillance, new treatments, and control of visceral leishmaniasis within the country. In light of this, the aim of this study was to isolate compounds from plants of the Caatinga biome, and to investigate their toxicity against promastigote and amastigote forms of Leishmania infantum chagasi, the main responsible parasite for South American visceral leishmaniasis, and evaluate their ability to inhibit acetylcholinesterase enzyme (AChE). A screen assay using luciferase-expressing promastigote form and an in situ ELISA assay were used to measure the viability of promastigote and amastigote forms, respectively, after exposure to these substances. The MTT colorimetric assay was performed to determine the toxicity of these compounds in murine monocytic RAW 264.7 cell line. All compounds were tested in vitro for their anti-cholinesterase properties. A coumarin, scoparone, was isolated from Platymiscium floribundum stems, and the flavonoids rutin and quercetin were isolated from Dimorphandra gardneriana beans. These compounds were purified using silica gel column chromatography, eluted with organic solvents in mixtures of increasing polarity, and identified by spectral analysis. In the leishmanicidal assays, the compounds showed dose-dependent efficacy against the extracellular promastigote forms, with an EC50 for scoporone of 21.4µg/mL, quercetin and rutin 26 and 30.3µg/mL, respectively. The flavonoids presented comparable results to the positive control drug, amphotericin B, against the amastigote forms with EC50 for quercetin and rutin of 10.6 and 43.3µg/mL, respectively. All compounds inhibited AChE with inhibition zones varying from 0.8 to 0.6, indicating a possible mechanism of action for leishmacicidal activity.

Nos últimos anos, o Ministério da Saúde do Brasil e a Organização Mundial da Saúde tem apoiado a investigação de novas tecnologias que possam contribuir para a vigilância, novos tratamentos e controle da leishmaniose visceral no país. Assim, o objetivo deste trabalho foi isolar compostos de plantas do bioma Caatinga, e investigar a toxicidade destes compostos contra as formas promastigotas e amastigotas de Leishmania infantum chagasi, principal parasita responsável pela leishmaniose visceral na América do Sul, e avaliar a sua capacidade para inibir a enzima acetil-colinesterase (AChE). Após a exposição aos compostos em estudo, foram realizados testes utilizando a forma promastigota que expressa luciferase e ELISA in situ para medir a viabilidade das formas promastigotas e amastigota, respectivamente. O ensaio colorimétrico MTT foi realizado para determinar a toxicidade destas substâncias utilizando células monocíticas murina RAW 264.7. Todos os compostos foram testados in vitro para as sua propriedade anti-colinesterásica. Um cumarina, escoparona, foi isolada a partir de hastes de Platymiscium floribundum, e os flavonóides, rutina e quercetina, foram isolados a partir de grãos de Dimorphandra gardneriana. Estes compostos foram purificados, utilizando cromatografia em coluna gel eluída com solventes orgânicos em misturas de polaridade crescente, e identificados por análise espectral. Nos ensaios leishmanicidas, os compostos fenólicos mostraram eficácia contra as formas extracelulares promastigotas, com EC50 para escoporona de 21.4µg/mL e para quercetina e rutina 26 e 30.3µg/mL, respectivamente. Os flavonóides apresentaram resultados comparáveis à droga controle, a anfotericina B, contra as formas amastigotas com EC50 para quercetina e rutina de 10.6 e 43.3µg/mL, respectivamente. Os compostos inibiram a enzima AChE com halos de inibição variando de 0,8 a 0,6cm, indicando um possível mecanismo de ação para a atividade leishmanicida.

Relaxant effect of the essential oil of Croton nepetifolius on ovine cervix

We investigated the in vitro effect of the essential oil of Croton nepetifolius Baill., Euphorbiaceae (EOCN), on spontaneous or induced contractions of circular and longitudinal muscles from the ovine cervix during the luteal phase of the estrous cycle. The relaxant effect of EOCN was expressed as a percentage of the contraction recorded before the addition of the oil and calculated relative to the preparations exposed only to the vehicle. The IC50 (concentration of oil required to produce a 50% maximal reduction in muscle contraction) for relaxation of spontaneous contractions in circular and longitudinal muscles was significantly lower than the IC50 for blockade of K+-induced contraction (27.19 µg mL-1 versus 262.72 µg mL-1 and 40.92 µg mL-1 versus 222.47 µg mL-1, respectively). Interestingly, there was a high degree of selectivity in the action of EOCN on cervix layers concerning the inhibition of acetylcholine-induced contraction in circular (IC50 277.10 µg mL-1) and longitudinal (IC50 52.56 µg mL-1) muscles. In conclusion, EOCN is able to relax ovine cervix during the luteal phase. This work opens the perspective of applying EOCN in ovine embryo transfer.

Water extracts of Brazilian leguminous seeds as rich sources of larvicidal compounds against Aedes aegypti L.

This study assessed the toxicity of seed water extracts of 15 leguminous species upon Aedes aegypti larvae. A partial chemical and biochemical characterization of water extracts, as well as the assessment of their acute toxicity in mice, were performed. The extracts of Amburana cearensis, Anadenanthera macrocarpa, Dioclea megacarpa, Enterolobium contortisiliquum and Piptadenia moniliformis caused 100 percent of mortalit y after 1 to 3 h of exposure. They showed LC50 and LC90 values ranging from 0.43 ± 0.01 to 9.06 ± 0.12 mg/mL and from 0.71 ± 0.02 to 13.03 ± 0.15 mg/mL, respectively. Among the secondary metabolite constituents, the seed water extracts showed tannins, phenols, flavones, favonols, xanthones, saponins and alkaloids. The extracts also showed high soluble proteins content (0.98 to 7.71 mg/mL), lectin (32 to 256 HU/mL) and trypsin inhibitory activity (3.64 = 0.43 to 26.19 = 0.05 gIT/kg of flour) The electrophoretic profiles showed a great diversity of protein bands, many of which already described as insecticide proteins. The extracts showed low toxicity to mice (LD50 > 0.15 = 0.01 g/kg body weight), but despite these promising results, further studies are necessary to understand the toxicity of these extracts and their constituentsfrom primary and secondary metabolism upon Ae. aegypti.

Este trabalho objetivou avaliar a toxicidade dos extratos aquosos de sementes de 15 espécies de leguminosas contra larvas de Aedes aegypti. Foi realizada uma caracterização química e bioquímica parcial dos extratos aquosos e a avaliação da toxicidade aguda em camundongos. Os extratos de Amburana cearensis, Anadenanthera macrocarpa, Dioclea megacarpa, Enterolobium contortisiliquum e Piptadenia moniliformis causaram 100 por cento de mortalidade depois de 1 a 3 h de exposição e mostraram valores de CL50 e CL90 entre 0,43 = 0,01 e 9,06 ± 0,12 e entre 0,71 = 0,02 e 13,03 = 0,15 mg/mL, respectivamente. Dentre os constituintes do metabolismo secundário, os extratos das sementes apresentaram taninos, fenóis, flavonas, flavonóis, xantonas, saponinas e alcalóides. Os extratos apresentaram alto teor de proteínas solúveis (0,98 to 7,71 mg/mL), lectina (32 to 256 UH/mL) e inibidor de tripsina (3,64 ± 0,43 to 26,19 = 0,05 gIT/kg de farinha). O perfil eletroforéticomostrou uma grande diversidade de proteínas, muitas dasquais já descritas como inseticidas. Os extratos mostraram baixa toxicidade ao camundongo (DL50 > 0,15 = 0,01 g/kg peso corporal), porém apesar desses resultados promissores, estudos posteriores são necessários para compreender a toxicidade desses extratos e de seus constituintes do metabolismo primário e secundário sobre Ae. aegypti.

Atividade inseticida in vitro do óleo de sementes de nim sobre Lutzomyia longipalpis (Diptera: Psychodidae) / In vitro insecticidal activity of seed neem oil on Lutzomyia longipalpis (Diptera: Psychodidae)

Lutzomyia longipalpis é o principal vetor da Leishmaniose visceral no Brasil. O objetivo deste trabalho foi avaliar o efeito do óleo de sementes de nim (Azadirachta indica) sobre ovos, larvas e adultos do vetor. Os insetos foram capturados no campo e mantidos no laboratório a ± 27 °C e 80 por cento de umidade relativa. Cinco tratamentos com diferentes concentrações foram realizados, usando-se dois controles negativos, um com água destilada e outro com Tween 80 (3 por cento) e um controle positivo com cipermetrina. Os ovos foram borrifados com o óleo em diferentes concentrações e avaliou-se o número de larvas eclodidas por 10 dias consecutivos. A mortalidade das larvas foi observada até a pupação, e a mortalidade dos adultos foi observada após 24, 48, e 72 horas. A análise estatística foi feita pelo teste de Tukey a 5 por cento de probabilidade. No teste com ovos, a maior concentração obteve 65,16 ± 3,24 por cento de eficácia. O teste com larvas apresentou 67,75 ± 2,21 por cento de eficácia na concentração de 100 mg.mL-1. Com adultos, a eficácia na concentração de 100 mg.mL-1 foi de 96,64 ± 4,11 por cento, após 24 horas. A análise fitoquímica revelou a presença de triterpenos. Esses resultados demonstram o potencial uso desse óleo no controle deste vetor.

Lutzomyia longipalpis is the main vector of visceral leishmaniasis in Brazil. The objective was to evaluate the effect of oil from (Azadirachta indica) neem seeds on eggs, larvae and adults of the vector. The insects were captured in the field and kept in the laboratory at ± 27 °C and 80 percent relative humidity. Five treatments with different concentrations were performed using two negative controls (distilled water and Tween 80) and a positive control. The eggs were sprayed with the oil at different concentrations and the number of hatched larvae evaluated for 10 days. Mortality of larvae was observed to pupation and adult mortality was observed after 24, 48, and 72 hours. Statistical analysis was performed by Tukey test at 5 percent probability. The highest oil concentration of eggs obtained 65.16 ± 3.24 percent efficacy for reducing egg hatching. The test with larvae showed 67.75 ± 2.21 percent efficacy at a concentration of 100 mg.mL-1. In adults, the efficacy of the 100 mg.mL-1 concentration was 96.64 ± 4.11 percent after 24 hours. The phytochemical analysis revealed the presence of triterpenes. These results demonstrate the potential use of this oil in the control of this vector.

Ação antioxidante de chás e condimentos de grande consumo no Brasil / Antioxidant action of teas and seasonings more consumed in Brazil

Antioxidantes são compostos que atuam inibindo e/ou diminuindo os efeitos desencadeados pelos radicais livres e compostos oxidantes. Os chás são bebidas populares e fontes significativas de compostos fenólicos, sendo considerados importantes integrantes das dietas devido às suas propriedades antioxidantes. O presente trabalho teve como objetivo avaliar a ação antioxidante de uma variedade de chás e condimentos mais consumidos no Brasil. Os chás analisados foram das plantas: Pneumus boldus Mold., Matricaria recutita L., Cymbopogon citratus (DC) Stapf, Baccharis trimera (Less.) DC, Camelia sinensis (L) O. Kuntze (fermentado), Camelia sinensis (não-fermentado), Lippia alba N.E. Brown,Mentha arvensis L., e Pyrus malus L. Os condimentos analisados foram: Eugenia aromatica Baill, Cinnamonum zeylanicum Blume, Laurus nobilis L. e Origanum vulgare L. O método utilizado para avaliar a ação antioxidante foi o da atividade seqüestradora de radicais livres DPPH em solução metanólica. Todas as amostras analisadas demonstraram atividade em suas diferentes concentrações. Camelia sinensis (não-fermentada) foi a mais ativa com CI50= 0,14 mg/mL, cujos principais compostos antioxidantes são epigalocatequinas. Os condimentos mais ativos foram Cinnamonum zeylanicum (CI50 = 0,37 mg/mL), Eugenia aromatica Baill (CI50 = 0,46 mg/mL) e Laurus nobilis (CI50 = 0,76 mg/mL), cujo principal antioxidante relatado foi o eugenol.

Antioxidants are compounds that remove free-radicals or minimize their availability to generate oxidative stress. Teas are popular beverages providing a significant source of phenolic compounds, important components of the human diet due to their antioxidant properties. The present work had the objective of evaluate the antioxidant action of teas and seasonings more consumed in Brazil. The analysed teas were from the plants: Pneumus boldus Mold., Matricaria recutita L., Cymbopogon citratus (DC) Stapf, Baccharis trimera (Less.) DC, Camelia sinensis (L) O. Kuntze (fermented), Camelia sinensis (not-fermented), Lippia alba N.E. Brown, Mentha arvensis L. and Pyrus malus L. The studied seasonings were: Eugenia aromatica Baill, Cinnamonum zeylanicum Blume, Laurus nobilis L. and Origanum vulgare L. The antioxidant activity was measured by the DPPH free radical scavenging method. All analysed samples demonstrated antioxidant activities. Camelia sinensis (not-fermented) (IC50= 0.14 mg/mL), was the most active and active principles are epicalocatechins. The stronger antioxidant seasonings were Cinnamonum zeylanicum (IC50 = 0.37 mg/mL), Eugenia aromatica Baill (IC50= 0.46 mg/ mL and Laurus nobilis (IC50 = 0.76 mg/mL), which presented as main antioxidant component eugenol.

Anthelmintic acetogenin from Annona squamosa L. Seeds

Annona squamosa seeds extracts showed anthelmintic activity against Haemonchus contortus, the main nematode of sheep and goat in Northeastern Brazil. A compound 1 was isolated from ethyl acetate extract and inhibited the egg hatching of H. contortus at 25 mg ml-1. The structure of 1 was determined as a C37 trihydroxy adjacent bistetrahydrofuran acetogenin based on spectroscopic analysis.

Extratos das sementes de Annona squamosa demonstraram atividade anti-helmíntica contra Haemonchus contortus, o principal nematódeo de ovinos e caprinos no Nordeste do Brasil. O composto 1 foi isolado do extrato acetato de etila e inibiu a eclosão dos ovos de H. contortus a 25 mg ml-1. A estrutura de 1 foi determinada como uma acetogenina C37 tri-hidroxi bis-tetrahydrofurano adjacente, baseando-se nos dados espectrais.