Phytochemical composition, antifungal activity, in vitro and in vivo toxicity of Syzygium cumini (L) Skeels leaves extract / Composición fitoquímica, actividad antifúngica, toxicidad in vitro e in vivo del extracto de hojas de Syzygium cumini (L) Skeels

This study determined phytochemical composition, antifungal activity and toxicity in vitro and in vivo of Syzygium cumini leaves extract (Sc). Thus, was characterized by gas chromatography coupled to mass spectrometry and submitted to determination of Minimum Inhibitory (MIC) and Fungicidal concentrations (MFC) on reference and clinical strains of Candida spp. and by growth kinetics assays. Toxicity was verified using in vitro assays of hemolysis, osmotic fragility, oxidant and antioxidant activity in human erythrocytes and by in vivo acute systemic toxicity in Galleria mellonella larvae. Fourteen different compounds were identified in Sc, which showed antifungal activity (MIC between 31.25-125µg/mL) with fungistatic effect on Candida. At antifungal concentrations, it demonstrated low cytotoxicity, antioxidant activity and neglible in vivotoxicity. Thus, Sc demonstrated a promising antifungal potential, with low toxicity, indicating that this extract can be a safe and effective alternative antifungal agent.

Este estudio determinó la composición fitoquímica, la actividad antifúngica y la toxicidad in vitro e in vivo del extracto de hojas de Syzygium cumini (Sc). Así, se caracterizó mediante cromatografía de gases acoplada a espectrometría de masas y se sometió a determinación de Concentraciones Mínimas Inhibitorias (CMI) y Fungicidas (MFC) sobre cepas de referencia y clínicas de Candida spp. y mediante ensayos de cinética de crecimiento. La toxicidad se verificó mediante ensayos in vitro de hemólisis, fragilidad osmótica, actividad oxidante y antioxidante en eritrocitos humanos y por toxicidad sistémica aguda in vivo en larvas de Galleria mellonella. Se identificaron catorce compuestos diferentes en Sc, que mostraron actividad antifúngica (CMI entre 31.25-125 µg/mL) con efecto fungistático sobre Candida. En concentraciones antifúngicas, demostró baja citotoxicidad, actividad antioxidante y toxicidad in vivo insignificante. Por lo tanto, Sc demostró un potencial antifúngico prometedor, con baja toxicidad, lo que indica que este extracto puede ser un agente antifúngico alternativo seguro y eficaz.

Cytotoxicity and antiviral activity evaluation of Cymbopogon spp hydroethanolic extracts

Cymbopogon citratus and C. nardus are noteworthy among the several existing plant species displaying medicinal properties, due to the potential pharmacological activity of these species, including antiviral, antibacterial, antifungal and anti-trypanosomal activities. The objective of this study was to carry out in vitro toxicity tests of plant extracts from both species and analyze potential antiviral activity against Human mastadenovirus serotype 5 (HAdV-5). Two cell lines (A549 and VERO) were used and mitochondrial and lysosomal viability were determined by the MTT and neutral red assay, respectively, after two exposure times (24 hours and six days). The aim of these assays was to counteract the behavior of the extracts against the different cell lines and determine their non-toxic concentration range, in order to evaluate possible antiviral activity against HAdV-5. Plaque reduction and inhibition index of viral titer assays were performed using the maximum non-cytotoxic concentrations (MNCC) of each extract. The results indicate MNCC at 625 µg/mL for all extracts, except for Cymbopogon nardus obtained with 80% ethanol (CN80), which showed toxicity at concentrations higher than 312.5 µg/mL. CN80 was the only extract that displayed potential activity against HAdV-5, at a concentration of 75 µg/mL, becoming a candidate for extract fraction purification and/or the isolation of substances related to the observed antiviral activity

Preliminary analysis of Brazilian Amazonian plant extracts using bench-top assays in a through-put basis aiming the identification of potential antioxidant natural product

Plants are a source of compounds that are used for the treatment of human and veterinary diseases. Brazil is one of the richest countries in the world in terms of biodiversity. The present study evaluated extracts using thin-layer chromatography to identify antioxidant activity and determine the presence of groups of compounds, in a large-scale basis. A total of 1,260 aqueous and organic extracts were obtained from plants that were collected in the Amazon and Atlantic rain forests. Thin-layer chromatography was performed to evaluate the presence of alkaloids, anthraquinones, cardioactive glicosides, terpenes, and phenolics and determine antioxidant and radical scavenging activity using the following reagents: Dragendorff's reagent, KOH, Kedde's reagent, sulphuric acid, NP reagent, ß-carotene, and 2,2-diphenyl-1-picrylhydrazyl (DPPH). Of the 1,260 extracts, 837 (66.43%) presented a ß-carotene/bleaching response, and 1,205 (95.63%) presented a radical scavenging response. Alkaloids were found in 203 extracts (16.11%). Anthraquinones were found in 14 extracts (1.11%), cardenolides were found in eight extracts (0.63%). The present findings shows the importance of the Brazilian Amazon plants as sources of antioxidant and radical scavenging active compounds.(AU)

Avaliação antimicrobiana de extratos vegetais e possível interação farmacológica in vitro / Interactive study of the antimicrobial activity of plant extracts

A atividade biológica de plantas medicinais tem sido o alvo de intensa investigação científica, constituindo uma importante fonte de novos produtos biologicamente ativos. O objetivo deste trabalho foi avaliar a atividade antimicrobiana interativa, in vitro, de seis extratos vegetais, sendo eles: cajueiro (Anacardium occidentale L.), barbatimão (Stryphnodendron adstringens (Mart.) Coville, aroeira (Myracrodruon urundeuva), favela [Cnidoscolus phyllacanthus (Müll. Arg.) Pax & Hoffm L.], fedegoso (Heliotropium indicum L.) e quixaba [Sideroxylon obtusifolium (Humb. ex Roem. & Schult.) TD Penn.], através do método de difusão em disco, frente às cepas padrão American Type Culture Collection (ATCC): Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922 e Pseudomonas aeruginosa ATCC 27853. Os resultados revelaram que dentre os extratos testados 50% deles apresentaram atividade frente à cepa de S. aureus ATCC 25923. Os extratos ativos foram cajueiro, barbatimão e aroeira. Os demais extratos não apresentaram atividade antimicrobiana frente às cepas em estudo. Quando se avaliou a ação interativa entre os extratos ativos, observou-se interferência antagônica, constatada pela redução dos diâmetros dos halo de inibição de crescimento bacteriano nas associações com os extratos de cajueiro/barbatimão, cajueiro/aroeira e barbatimão/aroeira. De modo que, o uso concomitante extratos vegetais merece um olhar muito cuidadoso pela possibilidade de ocorrer interferências entre eles, reduzindo a potência da atividade antimicrobiana, em relação a sua potência quando utilizados isoladamente.(AU)

The biological activity of medicinal plants has been the subject of intensive scientific research and is an important source of new biologically active products. The aim of this study was to evaluate the antimicrobial activity of six interactive plant extracts, which are: cajueiro (Anacardium occidentale L.), barbatimão (Stryphnodendron adstringens (Mart.) Coville, aroeira (Myracrodruon urundeuva), favela [Cnidoscolus phyllacanthus (Müll. Arg.) Pax & Hoffm L.], fedegoso (Heliotropium indicum L.) and quixaba [Sideroxylon obtusifolium (Humb. ex Roem. & Schult.) TD Penn.], by disk diffusion method, front to standard strains American Type Culture Collection (ATCC): Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853. The results revealed that among the tested extracts 50% of them showed activity against the strain of S. aureus ATCC 25923. The active extracts were cajueiro, barbatimão and aroeira. When we evalued the interative action between the active extracts, we observed an antagonistic interference, evidenced by the reduction of the diameter of inhibition zone of bacterial growth among the associations in the extracts of cajueiro/barbatimão, cajueiro/aroeira and barbatimão/aroeira. Therefore, concomitant use of plant deserves a careful look at the possibility of interference occurs between them, reducing the potency antimicrobial activity against its power when used alone.(AU)

Actividad tripanocida y citotóxica de extractos de plantas colombianas / Trypanocidal and cytotoxic activity of extracts of Colombian plants

Introducción. El tratamiento de la enfermedad de Chagas está basado en sólo dos medicamentos de eficacia limitada y con importantes efectos colaterales. La gran biodiversidad de la flora colombiana hace de la bioprospección una alternativa potencial en la búsqueda de nuevos antiparasitarios. Objetivo. Evaluar in vitro el potencial tripanocida y la citotoxicidad de extractos obtenidos de 23 plantas colombianas. Materiales y métodos. Se obtuvieron extractos de hojas, de tallos o de la planta entera, en solventes de diferente polaridad. La actividad contra epimastigotes y la citotoxicidad se evaluaron por el micrométodo enzimático con MTT. Los extractos activos contra epimastigotes y con baja citotoxicidad se evaluaron también en tripomastigotes y amastigotes intracelulares. Resultados. Se reporta la actividad tripanocida de 13 plantas colombianas y se confirma el efecto biológico de cuatro especies previamente evaluadas. Cuatro extractos activos en epimastigotes también fueron activos en tripomastigotes y, uno de ellos, en amastigotes. Este extracto fue aislado de la planta Hieronyma antioquensis, y presentó CI50 de 3,125, 11,48 y 2,85 µg/ml, e índices de selectividad de 25,7 y 27, para epimastigotes, tripomastigotes y amastigotes, respectivamente. Los resultados sugieren que este extracto es un candidato promisorio para el tratamiento de la enfermedad de Chagas. Conclusión. La flora colombiana es una fuente potencial de nuevas sustancias para la quimioterapia contra la enfermedad de Chagas. El micrométodo enzimático con MTT es una herramienta útil para la tamización de la actividad biológica en epimastigotes y posterior selección para ensayos con otros estadios del parásito.

Introduction. The treatment of Chagas disease is based on only two drugs with limited efficacy and significant side effects. The rich biodiversity of the Colombian flora makes bio-prospecting a potential alternative in the search for new antiparasitic drugs. Objective. Potential trypanocidal activity and cytotoxicity was assessed in extracts from 23 Colombian plants. Materials and methods. Extracts of leaves, stems, or of the whole plants were obtained in solvents of a range of polarities. The activity against Trypanosoma cruzi epimastigotes and the cytotoxicity were evaluated by the MTT enzymatic micro-method. Extracts active against epimastigotes and with lowcytotoxicity were also tested on trypomastigotes and intracellular amastigotes. Results. Among the extracts, biological activity was confirmed in 4 species. The extracts were active on epimastigotes and trypomastigotes; one was active also against amastigotes. The latter extract was isolated from the plant Hieronyma antioquensis and presented IC50 of 3.1 mg/ml for epimastigotes, 11.5mg/ml for trypomastigotes and 2.9 mg/ml for amastigotes. The selectivity indexes were 25, 7, and 27 respectively. Conclusions. The extract from H. antioquensis proved a promising candidate for Chagas disease treatment. Futhermore, the MTT enzymatic micromethod was a useful tool for screening biological activity on epimastigotes and other stages of the parasite for further extract trials.

Comparison of bioassays using the anostracan crustaceans Artemia salina and Thamnocephalus platyurus for plant extract toxicity screening / Comparação de bioensaios com os crustáceos Artemia salina e Thamnocephalus platyurus para abordagem de extratos de plantas com toxicidade

Three lethality bioassays, using the salt-water crustacean Artemia salina Leach, Artemiidae, (conventional 96 microwell plate test and the Artoxkit M microbiotest) and the freshwater crustacean Thamnocephalus platyurus Packard, Thamnocephalidae, (Thamnotoxkit F microbiotest), were compared using extracts of ten Guatemalan plant species. It was previously observed that five of them have anti-Artemia activity. These were: Solanum americanum Mill., Solanaceae, Gliricidia sepium (Jacq.) Kunth ex Walp., Fabaceae, Neurolaena lobata (L.) Cass., Asteraceae, Petiveria alliacea L., Phytolaccaceae, and Ocimum campechianum Mill., Lamiaceae. The five others: Curatella americana L., Dilleniaceae, Prunus barbata Koehne, Rosaceae, Quercus crispifolia Trel., Fagaceae, Rhizophora mangle L., Rhizophoraceae, and Smilax domingensis Willd., Smilacaceae, do not. All plants without anti-Artemia activity had no lethal effects in both assays with A. salina. For the plants with anti-Artemia activity the Artoxkit M was not sensitive to G. sepium and the conventional Artemia test was not sensitive to S. americanum, G. sepium and N. lobata. All the plant extracts, except for that of C. americana, had lethal effects on T. platyurus and the lethal median concentration (LC50) levels for this organism were in all cases substantially lower than those of the salt-water test species. This study revealed that T. platyurus is a promising test species worth further in depth investigation for toxicity screening of plant extracts with potential medicinal properties.

Três bioensaios de letalidade com o crustáceo de água salgada Artemia salina Leach, Artemiidae, (teste convencional em microplaca de 96 poós Artoxkit microbiotest M) e o crustáceo de água doce Thamnocephalus platyurus Packard, Thamnocephalidae (Thamnotoxkit microbiotest F), foram comparados utilizando extratos de dez espécies de plantas da Guatemala. Foi previamente observado que cinco delas possuem atividade anti-Artemia: Solanum americanum Mill., Solanaceae, Gliricidia sepium (Jacq.) Kunth ex Walp., Fabaceae, Neurolaena lobata (L.) Cass., Asteraceae, Petiveria alliacea L., Phytolaccaceae e Ocimum campechianum Mill., Lamiaceae. As outras cinco espécies, Curatella americana L., Dilleniaceae, Prunus barbata Koehne, Rosaceae, Quercus crispifolia Trel., Fagaceae, Rhizophora mangle L., Rhizophoraceae e Smilax domingensis Willd., Smilacaceae, não. Todas as plantas sem atividade anti-Artemia não tiveram nenhum efeito letal em ambos os ensaios com A. salina. Para as plantas com atividade anti-Artemia o M Artoxkit não foi sensível a G. sepium e teste convencional de Artemia não foi sensível a S. americanum, G. sepium e N. lobata. Todos os extratos vegetais, exceto o de C. americana, apresentaram um efeito letal sobre T. platyurus e a concentração letal média (CL50) para este organismo em todos os casos foram substancialmente inferiores aos da espécie de teste de água salgada. Este estudo revelou que T. platyurus é teste promissor para uma investigação aprofundada na seleção de extratos de plantas com potenciais propriedades medicinais.

Actividad larvicida de extractos vegetales acuosos en larvas de Aedes aegypti (primeros ensayos) / Larvicidal activity of aqueous plants extracts on Aedes aegypti larva (first trials)

El dengue es una enfermedad epidémica muy común en regiones tropicales y sub tropicales. La eliminación de criaderos y el control vectorial se encuentran entre las medidas en la lucha contra la enfermedad. Se han desarrollado diversas estrategias para mantener bajo índice poblacional del mosquito. Diversas investigaciones se han enfocado a la búsqueda de nuevos productos naturales, con actividad insecticida y larvicida, que puedan controlar la población de mosquitos, sin presentar riesgos al humano y animales domésticos. Realizamos una serie de bioensayos con extractos acuosos de plantas paraguayas, Annona muricata (chirimoya); Bulnesia sarmentoi (palo santo); Melia azederach (paraíso); Zanthoxylum chiloperone var. Angustifolium (tembetary hú) y Bixa orellana (urukú), para comprobar en cada planta, su actividad y eficacia como larvicida, contralarvas del mosquito Aedes agypti. Dichas larvas, fueron colectadas de diversas zonas de Asunción y el Gran Asunción, durante la epidemia de fiebre amarilla del año 2007. Las semillas de la Annona mucricata (chirimoya), presentaron una buena actividad larvicida, ya que a la mínima concentración del 5%, han tenido un efecto mortal para las larvas, comparable al observado en los controles positivos (que contenían temefos 1%). En cambio, M. aezsederach (paraíso) y Z. chiloperone (tembetary hú) no mostraron actividad larvicida a esa dosis, ni aún a otras superiores. Por otro lado B. sarmientoi (palo santo) y B. orellana (urukú), presentaron cierto efecto larvicida, eliminando al 18% de larvas a las 72 horas post-exposición. Se observó una marcada diferencia de actividad, entre el extracto de semillas chirimoya con los demás extractos probados

Dengue is an common epidemic disease in tropical and subtropical regions. The elimination of breeding sites and vector control are among the most widely used measures in the fight against the disease. Many strategies have been developed to keep low rates of mosquito populations. Several research studies have been focused on finding natural products with insecticide and larvicide activity that could effectively control these mosquito populations without risks for the human populations and domestic animals. In this work, we have performed a series of bioassays with aqueous extracts of Paraguayan plants: Annona muricata (cherimoya), Bulnesia sarmentoi (palo santo), Melia azederach (paradise), Zanthoxylum chiloperone var. Angustifolium (tembetary hú) and Bixa orellana (uruku) in order to check the effectiveness and activity as larvicide of each plant gainst Aedes aegypti larvae. The larvae were collected in various areas of Asuncion and Great Asuncion during the yellow fever outbreak of 2007. The seeds of A. mucricata (cherimoya or custard apple) showed good larvicidal activity, i.e. at the minimum concentration of 5%, showed lethality against larvae comparable to that observed in positive controls (containing 1% temephos). On the other hand, M. azederach (paradise) and Z. chiloperone (tembetary hú) did not show any larvicidal activity at the same dose and even at higher doses. B. sarmientoi (palo santo) and Bixa orellana (uruku) showed some larvicidal effect killing larva (18%) at 72 hours post-exposure. There was a marked difference in activity between the cherimoya seeds extract and the other extracts tested

Efeitos de extratos de plantas na biologia de Spodoptera frugiperda (J. E. Smith, 1797) (Lepidoptera: Noctuidae) mantida em dieta artificial / Effects of plant extracts on the biology of Spodoptera frugiperda (J. E. Smith, 1797) (Lepidoptera: Noctuidae) maintained under artificial diet

Avaliaram-se os efeitos dos extratos aquosos a 10 por cento de folhas e ramos de arruda (Ruta graveolens L.), folhas e ramos de melão-de-são-caetano (Momordica charantia L.), folhas do alecrim-pimenta (Lippia sidoides Cham.) e fruto verde de mamona (Ricinus communis L.), sobre a biologia da lagarta-do-cartucho do milho (Spodoptera frugiperda), mantida em dieta artificial. Os parâmetros avaliados foram duração e viabilidade das fases larval e pupal, peso de pupa, fecundidade, fertilidade e longevidade de adultos. Larvas de S. frugiperda recém-eclodidas foram colocadas em tubos de ensaio com dieta artificial, contendo os extratos de cada material testado. O extrato aquoso do fruto verde de R. communis apresentou bioatividade, nos parâmetros duração larval e pupal e peso de pupa. O extrato aquoso de R. graveolens reduziu o peso de pupa. A dieta contendo extrato de folhas e ramos de M. charantia reduziu a viabilidade larval e o peso de pupa. O extrato aquoso de folhas de L. sidoides não afetou as fases larval e pupal, reduziu a postura e a viabilidade de ovos e aumentou a longevidade de adultos de S. frugiperda. A viabilidade de pupa não foi afetada pelos extratos testados.

The effects of aqueous extracts, at 10 percent concentration of leaves and branches of Ruta graveolens L., leaves and branches of Momordica charantia L., leaves of Lippia sidoides Cham. and green fruits of Ricinus communis L. were evaluated on the biology of fall armyworm (Spodoptera frugiperda) maintained under artificial diet. The evaluated parameters were: duration and viability of the larval and pupal phases, pupa weight, fecundity, fertility and longevity of adults. Just-hatched larvae of S. frugiperda was placed in test tube with artificial diet containing extracts of each tested material. The aqueous extract of the green fruits of R. communis presented bioactivity upon duration and weight of larval and pupal phases. The aqueous extract of R. graveolens reduced weight of pupa. The diet containing extract of M. charantia reduced larval viability and weight of pupa. The aqueous extract of L. sidoides did not affect larval and pupal phases, but reduced the posture and the viability of eggs and increased longevity of adults of S. frugiperda. The viability of pupa was not affected by the extracts.

Actividad larvicida de extractos vegetales acuosos en larvas de Aedesaegypti: primeros ensayos / Larvicidal activity of aqueous plants extracts on Aedes aegypti larva: first trials

El dengue es una enfermedad epidémica muy común en regiones tropicales y subtropicales. La eliminación de criaderos y el control vectorial se encuentran entre las medidas en la lucha contra la enfermedad. Se han desarrollado diversas estrategias para mantener bajo índice poblacional del mosquito. Diversas investigaciones se han enfocado a la búsqueda de nuevos productos naturales, con actividad insecticida y larvicida, que puedan controlar la población de mosquitos, sin presentar riesgos al humano y animales domésticos. Realizamos una serie de bioensayos con extractos acuosos de plantas paraguayas, Annona muricata (chirimoya); Bulnesia sarmentoi (palo santo); Melia azederach (paraíso); Zanthoxylum chiloperone var. Angustifolium (tembetary hú) y Bixaorellana (urukú), para comprobar en cada planta, su actividad y eficacia como larvicida, contra larvas del mosquito Aedes agypti. Dichas larvas, fueron colectadas de diversas zonas de Asunción y el Gran Asunción, durante la epidemia de fiebre amarilla del año 2007. Las semillas de la Annona mucricata (chirimoya), presentaron una buena actividad larvicida, ya que a la mínima concentración del 5%, han tenido un efecto mortal para las larvas, comparable al observado en los controles positivos (que contenían temefos 1%). En cambio, M. aezsederach (paraíso) y Z. chiloperone (tembetary hú) no mostraron actividad larvicida a esa dosis, ni aún a otras superiores. Por otro lado B. sarmientoi (palo santo) y B. orellana (urukú), presentaron cierto efecto larvicida, eliminando al 18% delarvas a las 72 horas post-exposición. Se observó una marcada diferencia de actividad, entre el extracto de semillas chirimoya con los demás extractos probados.

Dengue is an common epidemic disease in tropical and subtropical regions. Theelimination of breeding sites and vector control are among the most widely usedmeasures in the fight against the disease. Many strategies have been developed to keep low rates of mosquito populations. Several research studies have been focused on findingnatural products with insecticide and larvicide activity that could effectively control these mosquito populations without risks for the human populations and domestic animals. In this work, we have performed a series of bioassays with aqueous extracts of Paraguayan plants: Annona muricata (cherimoya), Bulnesia sarmentoi (palo santo), Melia azederach (paradise), Zanthoxylum chiloperone var. Angustifolium (tembetary hú) and Bixa orellana(uruku) in order to check the effectiveness and activity as larvicide of each plant gainst Aedes aegypti larvae. The larvae were collected in various areas of Asuncion and Great Asuncion during the yellow fever outbreak of 2007. The seeds of A. mucricata (cherimoya or custard apple) showed good larvicidal activity, i.e. at the minimum concentration of 5%, showed lethality against larvae comparable to that observed in positive controls(containing 1% temephos). On the other hand, M. azederach (paradise) and Z. chiloperone (tembetary hú) did not show any larvicidal activity at the same dose and even at higher doses. B. sarmientoi (palo santo) and Bixa orellana (uruku) showed somelarvicidal effect killing larva (18%) at 72 hours post-exposure. There was a marked difference in activity between the cherimoya seeds extract and the other extracts tested.