ABSTRACT
Abstract Fridericia caudigera and Cuspidaria convoluta (Bignoniaceae) species, which grow in the northwest of Argentina, have shown antibacterial effect against strains isolated from skin infections, and each one displayed synergism with commercial antibiotics. The aims of this work were to evaluate the antibacterial activity and toxicity of the combination of these two plant species, and to design a stable gel for topical use including the blend of extracts. The combination of extracts was evaluated for synergistic effects (chequerboard assay), genotoxicity (Ames test) and cytotoxicity (Artemia salina test). A gel was subsequently formulated with the combination of extracts using carboxymethylcellulose as a polymer. The following physico- chemical characteristics of the gel formulation: pH, viscosity, spreadability and total phenol content, as well as resistance to severe temperature changes, biological activity (diffusion in agar), in vitro permeation (Franz cells) and primary dermal irritation (Draize test) were analyzed. The combination of extracts showed a synergistic effect on pathogenic bacteria and was not toxic in the in vitro tests. The gel was stable and retained the antimicrobial activity of the original extracts. The formulation proposed in this work could constitute an alternative for primary skin infections since it proved to be safe for topical administration.
Subject(s)
Plants/adverse effects , Artemia/classification , Skin/injuries , Bignoniaceae/classification , In Vitro Techniques/methods , Anti-Bacterial Agents/pharmacology , Mutagenicity Tests/instrumentationABSTRACT
RESUMO A cada dia, cepas bacterianas estão tornando-se resistentes a diversos antibióticos, o que faz necessária a busca de novas substâncias eficazes para o tratamento de doenças. Desta forma, este trabalho reporta o estudo preliminar toxicológico, antibacteriano e fitoquímico do extrato etanólico das folhas de Jatropha mollissima (pinhão-bravo, Euphorbiaceae), coletada no Município de Tauá, Ceará, Nordeste Brasileiro. Inicialmente, realizou-se o teste de toxicidade do extrato contra Artemia salina. Na sequencia, foi realizado o ensaio antibacteriano contra quatro cepas bacterianas Gram-negativas (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Hafnia alvei ATCC 51873, Klebsiella pneumoniae ATCC 13883) e uma cepa Gram-positiva (Enterococcus faecalis ATCC 29212). Finalmente, fez-se a análise fitoquímica preliminar do extrato ativo para detecção das principais classes de metabólitos especiais. Como resultado, o extrato etanólico das folhas de J. mollissima se mostrou tóxico para Artemia salina, pois apresentou CL50 igual a 406,02 μg/mL. Quanto à ação antibacteriana, o extrato se mostrou ativo contra a bactéria Gram-positiva Enterococcus faecalis ATCC 29212, apresentando moderada atividade antibacteriana (halo de inibição igual a 7,03 mm). Evidenciou-se no extrato bioativo a presença de cumarinas, fenóis, taninos, flavonoides (flavonóis e flavanonas), alcaloides e esteroides, ambas as classes reportadas como antimicrobianos. Portanto, esse extrato tem potencial para ser usado na produção de fármacos contra infecções causadas por bactérias Gram-positivas. No entanto, as informações direcionam estudos futuros para o isolamento e identificação dos compostos bioativos, monitorados sob a ação antibacteriana mais expressiva.
ABSTRACT Each day, bacterial strains are becoming more resistant to various antibiotics, which requires the search for new effective substances for the treatment of diseases. Thus, this study reports the toxicological, antibacterial, and phytochemical preliminary study of the ethanolic extracts of Jatropha mollissima (pinhão-bravo, Euphorbiaceae) leaves, collected in Tauá, Ceará, Northeast of Brazil. Initially, we performed the toxicity testing of the extract against Artemia salina. Then, we conducted the antibacterial assay against four Gram-negative bacterial strains (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Hafnia alvei ATCC 51873, Klebsiella pneumoniae ATCC 13883), and one Gram-positive strain (Enterococcus faecalis ATCC 29212). Finally, we carried out the preliminary phytochemical analysis of the active extract to detect the main classes of special metabolites. As a result, the ethanolic extract of J. mollissima leaves was toxic to Artemia salina, because it presented LC50 equal to 406.02 µg/mL. Regarding antibacterial action, the extract was active against the Gram-positive bacteria Enterococcus faecalis ATCC 29212, with moderate antibacterial activity (inhibition zone equal to 7.03 mm). The bioactive extract had the presence of coumarins, phenols, tannins, flavonoids (flavanols and flavonones), alkaloids and steroids, both classes reported as antimicrobials. Therefore, this extract has the potential to be used in the production of drugs against infections caused by Gram-positive bacteria. However, these information require further studies for the isolation and identification of bioactive compounds, monitored under the more expressive antibacterial action.
Subject(s)
Toxicity Tests/methods , Euphorbiaceae/classification , Anti-Bacterial Agents , Artemia/classification , Enterococcus faecalis/classificationABSTRACT
RESUMO A dengue está entre as doenças virais de propagação vetorial mais importante no mundo, causando sérios impactos de morbidade e mortalidade. Desta forma, o presente trabalho teve como objetivo analisar a composição química e a toxicidade do óleo essencial de Myrcia sylvatica (G. Mey) D.C. frente Aedes aegypti e Artemia salina. Folhas de M. sylvatica foram coletadas no Parque Nacional da Chapada das Mesas, no município de Carolina (MA) no mês de fevereiro de 2012. O óleo foi obtido por hidrodestilação e sua composição química foi determinada por cromatografia gasosa acoplada à espectrometria de massa (CG/EM). O bioensaio frente Artemia salina e às larvas de 3° estádio de Aedes aegypti foram realizados em diferentes concentrações. Os dados de mortalidade foram avaliados por regressão linear para determinar os valores de CL50. Obteve-se 0,5% de rendimento, sendo o (E)-cariofileno o constituinte majoritário. O óleo essencial apresentou uma CL50 = 79,44 µg/mL frente A. salina, sendo considerado altamente tóxico. No entanto, este óleo não demonstrou efeito sobre as larvas de A. aegypti. Considerando que o teste de Artemia salina tem correlação com atividades biológicas de grande interesse terapêutico como antitumoral, o óleo essencial das folhas de M. sylvatica demonstrou potencial para desenvolvimento de produtos farmacêuticos.
ABSTRACT Dengue is among the most important viral diseases of vector spread in the world, causing serious impacts of morbidity and mortality. Thus, this study aimed to analyze the chemical composition and toxicity of the Myrcia sylvatica (G. Mey.) DC essential oil against Aedes aegypti and Artemia salina Leach. Leaves of M. sylvatica. were collected in the Tables Chapada National Park, in the municipality of Carolina (MA) in february 2012. The oil was obtained by hydrodistillation and their chemical composition was determined by GC / MS. The bioassay front and Artemia salina larvae of the 3° stage of Aedes aegypti were performed at different concentrations. Mortality data were evaluated by linear regression to determine the LC50. Was obtained 0.5% yield values, and (E) -caryophyllene the major constituent. The essential oil showed a LC50 = 79.44 mg / mL front A. saline and is considered highly toxic. However, this oil showed no effect on larvae of A. aegypti. Whereas the brine shrimp test has regard to several biological activities, as antitumoral property, the essential oil from the leaves of M. sylvatica demonstrated potential for development of medicines.
Subject(s)
Toxicity , Oils, Volatile/chemistry , Chemistry , Aedes/classification , Myrtaceae/chemistry , Artemia/classification , LarvicidesABSTRACT
RESUMOA espécie Dicksonia sellowiana, conhecida popularmente como xaxim, é uma samambaia natural do continente americano e encontrada no Brasil na Mata Atlântica. Em 2001 foi inserida na lista do IBAMA como espécie ameaçada de extinção em decorrência da exploração para a confecção de vasos para a jardinagem. O presente trabalho descreve o potencial antioxidante lipídico (TBARS e Sistema β-caroteno/ácido linoleico) do extrato bruto e frações obtidos através de aparato de Soxhlet de frondes de Dicksonia sellowiana, além da atividade citotóxica e hemolítica in vitro. Pelo método TBARS, todas as amostras testadas apresentaram atividade, destacando a fração acetato de etila e extrato bruto cuja atividade foi comparável ao padrão ácido ascórbico. No sistema β-caroteno/ácido linoleico, a fração acetato de etila e extrato bruto apresentaram inibição da oxidação do ácido linoleico, destaque para a fração acetato de etila que não se diferenciou estatisticamente do padrão BHT. Na avaliação da toxicidade preliminar, não fora observado atividade citotóxica e hemolítica do extrato bruto e frações nos modelos testados. Os resultados demonstram o potencial antioxidante da espécie vegetal nos modelos de inibição da oxidação lipídica sem apresentar toxicidade.
ABSTRACTThe species Dicksonia sellowiana, popularly known as tree fern, are a natural fern from American continent found in the Atlantic Forest in Brazil. In 2001 it was added to the list of IBAMA as endangered species due to its exploitation for pots craft for gardening species. This paper describes the antioxidant lipid potential (TBARS and System β-carotene / linoleic acid) of the crude extract and fractions obtained by Soxhlet apparatus of Dicksonia sellowianafronds, besides the cytotoxic and hemolytic activity in vitro. By the TBARS method, all samples tested showed activity, particularly the fraction ethyl acetate and the crude extract whose activity was comparable to standard ascorbic acid. In the system β-carotene / linoleic acid, the fraction of ethyl acetate and crude extract inhibited the oxidation of linoleic acid, especially the ethyl acetate fraction which did not differ statistically from the standard BHT. In a preliminary assessment of toxicity,cytotoxic and hemolytic activity of crude extracts and fractions in the tested models were not observed. The results of this study indicate the antioxidant potential of this plant species in models of inhibiting lipid oxidation without producing toxicity.
Subject(s)
Lipid Peroxidation , Tracheophyta/classification , Antioxidants/analysis , Artemia/classification , Toxicity/methodsABSTRACT
RESUMO A espécie Senecio westermanii Dusén pertencente à família Asteraceae é planta endêmica e nativa do Brasil encontrada na região da Floresta Atlântica, nos estados do Paraná e São Paulo. O objetivo deste trabalho foi avaliar a composição fitoquímica do extrato bruto etanólico e frações das partes aéreas (folha e caule) de S. westermanii utilizando-se a análise fitoquímica qualitativa e cromatografia líquida de alta eficiência com detector de arranjo de diodos (CLAE-DAD), avaliar in vitro a toxicidade preliminar utilizando Artemia salina e o potencial antioxidante. O estudo fitoquímico qualitativo revelou a presença de alcaloides, flavonoides, iridoides, esteroides/triterpenos, heterosídeos saponínicos e aminogrupos. Através da análise realizada por CLAE-DAD obteve-se o fingerprint característico de cada amostra. No ensaio frente à A. salina houve ausência de toxicidade das amostras, o resultado da DL50 para todas as amostras foi superior a 1000 μg/mL. Todas as amostras apresentaram atividade antioxidante pela redução do complexo fosfomolibdênio, com destaque para a fração clorofórmio que apresentou atividade antioxidante de 92,51% em relação ao padrão rutina. Com relação ao ensaio de redução do radical DPPH•, a fração acetato de etila apresentou IC50 de 26,98 μg/mL. Pelo ensaio do ácido tiobarbitúrico (TBARS) a fração hexano apresentou o melhor índice antioxidante em relação ao padrão rutina. Os resultados obtidos demonstram evidências de que a espécie é fonte potencial de antioxidantes naturais, estimulando assim novos estudos que viabilizam sua utilização no tratamento de patologias associadas aos radicais livres. Além disso, a espécie não apresentou atividade tóxica preliminar, assegurando sua aplicabilidade.
ABSTRACT The species Senecio westermanii Dusén, which belongs to the Asteraceae family, is an endemic and native plant from Brazil. It is found mainly in the Atlantic Forest region, in Paraná and São Paulo states. This study aimed to evaluate the phytochemical composition using a qualitative phytochemical analysis and high performance liquid chromatography with diode array detector (HPLC-DAD), to evaluate preliminary toxicity in vitro using Artemia salina and antioxidant potential. Qualitative phytochemical analysis revealed the presence of alkaloids, flavonoids, iridoids, steroids, triterpenes, saponinic glycosides and aminogroups. The analysis by HPLC-DAD provided the characteristic fingerprint of each sample. In the A. salina assay, the results of LD50 over 1000 μg/mL for all samples did not indicate toxicity of the evaluated extracts. All samples demonstrated antioxidant activity by reducing phosphomolybdenum complex. The highest activity was detected in the chloroform fraction, which presented activity of 92.51% compared to the standard rutin. Regarding the trial to reduce the radical DPPH•, the ethyl acetate fraction showed an IC50 of 26.98 µg/mL. In the thiobarbituric acid assay (TBARS), the hexane fraction showed the highest antioxidant activity compared to the standard rutin. These findings indicate that the Senecio westermanii is a potential source of natural antioxidants, stimulating new studies that enable their use in the treatment of disorders associated with free radicals. In addition, the species did not present toxicity in its evaluation, ensuring its applicability.
Subject(s)
Artemia/classification , Toxicity , Asteraceae/chemistry , Antioxidants/pharmacology , Chromatography, High Pressure Liquid/instrumentationABSTRACT
BACKGROUND: Actinomycetes are gram positive bacteria with high G + C content in their DNA and are capable of producing variety of secondary metabolites. Many of these metabolites possess different biological activities and have the potential to be developed as therapeutic agents. The aim of the present study was to screen actinomycetes inhabiting halophilic environment such as Khewra salt mines present in Pakistan for cytotoxic and antitumor compounds. RESULTS: An actiomycetes strain designated as Streptomyces sp. KML-2 was isolated from a saline soil of Khewra salt mines, Pakistan. The strain Streptomyces sp. KML-2 showed 84 % cytotoxic activity against larvae of Artemiasalina. In the screening phase, the strain exhibited significant antitumor activity with IC50 values of 12, 48 and 56 µg/ml against Hela, MDBK and Vero cell lines, respectively. After that extract from 20 l fermentation was used to purify secondary metabolites by several chromatographic techniques. Structure elucidation of isolated compounds revealed that it is highly stable producer of Chromomycin SA (1) and 1-(1H-indol-3-yl)-propane-1,2,3-triol (2). Both of the isolated compounds showed significant antitumor activity against Hela and MCF-7 cancer cell lines (IC50 values 8.9 and 7.8 µg/ml against Hela; 12.6 and 0.97 µg/ml against MCF-7, respectively). The 16S rRNA gene sequence (1437 bp) of the strain confirm its identity (99 %) with Streptomyces griseus. CONCLUSIONS: From this research work we were successful in isolating two potent antitumor compounds, Chromomycin SA and 1-(1H-indol-3-yl)-propane-1,2,3-triol from Streptomyces KML-2 strain, isolated from Khewra salt mine. As such this is the second report which confirms that S. griseus can produce Chromomycin SA without introducing any mutagenesis in its biosynthesizing gene cluster and isolated indole derivative is being reported first time from any member of actinomycetes group with having novel antitumor activity against Hela and MCF-7 cells Nucleotide sequences: Nucleotide sequence data reported are available in the GenBank database under the accession number: GenBank KJ009562.
Subject(s)
Humans , Animals , Cattle , Soil Microbiology , Streptomyces/chemistry , Antineoplastic Agents/pharmacology , Pakistan , Phylogeny , Artemia/classification , Artemia/drug effects , Salts , Soil/chemistry , Streptomyces/isolation & purification , Streptomyces/ultrastructure , Streptomyces griseus/classification , Tetrazolium Salts , Vero Cells , RNA, Ribosomal, 16S/genetics , Chromomycins/classification , Chromomycins/pharmacology , HeLa Cells , Microscopy, Electron, Scanning , Cell Line , Chlorocebus aethiops , Chromatography/methods , Sequence Analysis, RNA , Inhibitory Concentration 50 , MCF-7 Cells , Formazans , Glycerol/analogs & derivatives , Glycerol/pharmacology , Larva/drug effects , Mining , Anti-Bacterial Agents/isolation & purification , Anti-Bacterial Agents/pharmacology , Antineoplastic Agents/isolation & purificationABSTRACT
Avaliou-se a expressão de ácidos graxos, macronutrientes e colesterol de camarões-da-malásia alimentados com dieta adicionada de sementes de linhaça. Quinhentos camarões foram distribuídos em dois grupos e alimentados durante 90 dias com dietas contendo: 1) lipídeo proveniente de linhaça e 2) óleo de soja (controle). A concentração de proteína foi maior (Р<0,05) para o grupo-linhaça, 15,8±0,09%, em relação ao grupo-controle, 15,2±0,04%. O peso corporal também se apresentou maior no grupo-linhaça, 1,38±0,93g, em relação ao grupo-controle, 0,93±0,48g. O grupo-linhaça apresentou teor de colesterol de 120,8±23,9mg/100g e o grupo-controle, 130,2±13,9mg/100g, porém sem diferença estatística significativa. Em relação ao perfil lipídico, a suplementação da dieta com linhaça influenciou a incorporação do ácido oleico (C18:1, ω-9) e melhorou a relação ω-6/ω-3 na carne do camarão (Р<0,05). Concluiu-se que a adição de linhaça à ração promoveu importantes modificações na qualidade nutricional dos tecidos do camarão.
The aim of the present study was to analyze the chemical composition of shrimp fed with a diet added of linseed flour. Five hundred post larvae shrimp were divided into two groups: a linseed group fed with a diet based on casein complemented with linseed and a control group, fed a diet without linseed flour. The cholesterol content for the linseed group was 120.8±23.9 mg/100g while for the control group it was 130.2±13.9 mg/100g. However, the protein content of 15.8±0.09 % and the body weight of 1.38±0.93g for the linseed group were higher than for the control group. It was concluded that the addition of linseed to the diet seems to increase the protein, weight and oleic acid (C18:1, ω-9) in the shrimp tissue. The ratio of polyunsaturated fatty acids/saturated fatty acids and ω-6/ω-3 for the linseed group was more suitable than for the control group.
Subject(s)
Animals , Cholesterol , Flour/analysis , Nutrients , Artemia/classificationABSTRACT
O presente estudo teve como objetivo uma citotoxicidade sobre Artemia salina de vinhos e dois extratos de cinco espécies do gênero Lychnophora e de uma espécie de Lychnophoriopsis . Os extratos solubilizados em DMSO, preparados nas concentrações finais de 100, 250, 375, 500 e 600 µg mL -1 , foram adicionados a recipientes contendo náuplios de Artemia salina (10 unidades cada) e completo em volume para 5 mL de solução marinha. Lapachol e DMSO 5% foram como controles positivo e negativo, respectivamente. Como amostras foram mantidas sob iluminação e como larvas mortas foram contadas após 24 horas de contato. O cálculo da LC 50 foi feito com o programa Probitos. Os extratos brutos etanólicos de cinco Espécies apresentaram baixa letalidade nd Seguinte Ordem: Lychnophora trichocarpha (LC 50 = 672,38 ng mL -1 )> Lychnophora pinaster (LC 50 = 678,73 ng mL -1 )> Lychnophora ericoides (LC 50 = 738,09 µg mL -1 )> Lycellophoriopsis candelabro (LC 50 = 812,57 µg mL -1 )> Lychnophora passerina (LC 50 = 921,78 µg mL -1 ). Todos os extratos testados de Lychnophoriopsis candelabro eo extrato clorofórmico de Lychnophora staavioides mostraram leve toxicidade sobre A. salina . Os resultados indicaram que existem substâncias com potencial atividade farmacológica em todas as espécies testadas.
The present study aimed to evaluate on Artemia salina the citotoxicity of twenty-two extracts from five species of the genus Lychnophora and one species of the genus Lychnophoriopsis. The extracts solubilized in DMSO and prepared at the final concentrations of 100, 250, 375, 500 and 600 ïg mL-1 were added to tubes containing Artemia salina nauplii (10 units each) and filled to 5 mL total volume with artificial salt water. Lapachol and 5% DMSO were used as positive and negative controls, respectively. The samples were kept under light and dead larvae were counted after 24 hours of contact. LC50 was calculated by using Probit software. The crude ethanol extracts from five species showed low lethality in the following order: Lychnophora trichocarpha (LC50 = 672.38 ïg mL-1) > Lychnophora pinaster (LC50 = 678.73 ïg mL-1) > Lychnophora ericoides (LC50 = 738.09 ïg mL-1) > Lychnophoriopsis candelabrum (LC50 = 812.57 ïg mL-1) > Lychnophora passerina (LC50 = 921.78 ïg mL-1). All tested extracts from L. candelabrum and chloroform extract from L. staavioides showed light toxicity on A. salina. Results indicated that there are substances with potential pharmacological activity in all tested species
Subject(s)
Artemia/classification , Biological Assay/instrumentation , Asteraceae/classificationABSTRACT
Cysts of Artemia spp. collected from February 1997 to February 2000 in the Real de Salinas solar saltworks, Campeche, Mexico, were compared with Artemia franciscana (batch number 8,131 Microfeast Artemia Cysts, Texas, USA). The variables determined in these two populations were: number of cysts per gram, hatching percentage, hatching efficiency, hatching rate, hatching synchrony and hatching biomass, as well as diameter of the cysts and length of the nauplii (instar I). For Salinas, the average diameters of the encapsulated and decapsulated cysts were 230.5 +/- 4.14 and 221.8 +/- 3.39 microm, respectively. The thickness of the cyst shell was 4.35 +/- 0.68 microm and the length of the nauplii was 388.11 +/- 4.39 microm, this last value is among the smallest reported in the literature. For the commercial population of A. franciscana, the average diameters of the encapsulated and decapsulated cysts were 230.21 +/- 12.49 and 216.96 +/- 13.71, respectively. With respect to the corion thickness and length of the nauplii the values were 6.62 +/- 2.72 and 424.70 +/- 30.08, respectively. The protein value of the cysts (47.91 %) and nauplii (50.5 %) of Artemia population from Real de Salinas, are considered adequate to be used as food in aquaculture. The results indicate that the population from Real de Salinas presents positive features for its use in aquaculture in the region.
Se utilizaron quistes de Artemia recolectados en períodos de febrero de 1997 a febrero del 2000 en las salinas de Real de Salinas, Campeche, México y como especie de comparación Artemia franciscana (lote No. 8131 Microfeast Artemia Cysts, Texas, EEUU.). Se determinó el número de quistes por gramo, porcentaje de eclosión (%H), eficiencia, tasa, sincronía y biomasa de eclosión, así como diámetro de quistes y longitud de los nauplios (instar I) de ambas poblaciones. Para la población de Real de Salinas, el diámetro de quistes capsulados y descapsulados en promedio fue de 230.5±4.14 y 221.8±3.39 [mi]m, respectivamente. El grosor del corion de 4.35±0.68 [mi]m y la longitud del nauplio de 388.11±4.39 [mi]m, siendo este último de los más pequeños que se hayan publicado en la literatura. Para la población de A. franciscana comercial, el diámetro del quiste capsulado y descapsulado presentó valores promedio de 230.21±12.49 y 216.96±13.71 [mi]m, respectivamente. Respecto al grosor del corion y longitud del nauplio, los valores fueron de 6.62±2.72 y 424.70±30.08 [mi]m, respectivamente. El valor proteico de los quistes (47.91 %) y nauplios (50.5 %) de Artemia de la población de Real de Salinas, se considera adecuado para uso como alimento en acuicultura. La población de Real de Salinas presenta bondades para su aprovechamiento en la acuicultura regional.