Compuestos fenólicos de Tagetes lucida Cav. con efecto antibacteriano por daños a la membrana / Phenolic compounds of Tagetes lucida Cav. with antibacterial effect due to membrane damage

Tagetes lucida Cav. (Asteraceae = Compositae) se usa para tratar infecciones estomacales. El estudio se centró en evaluar la composición y el efecto antimicrobiano de un extracto de T. lucida Cav. La planta se extrajo con etanol al 10% p/v, y la composición del extracto se analizó por Rp-HPLC-MS. El efecto antibacteriano se evaluó contra Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa y Salmonella choleraesuis utilizando métodos de difusión por disco, microdilución y bioautografía. Los ensayos de sytox y cometa fueron utilizados para evaluar el mecanismo de acción. De esta forma, se detectaron nueve compuestos fenólicos en el extracto de T. lucida. El extracto exhibió actividad solo en S. aureus (MIC de 4.000 mg/ml). La bioautografía reveló que los compuestos fenólicos podrían actuar sinérgicamente. Las pruebas de sytox y cometa mostraron una acción antibacteriana del extracto sobre la membrana bacteriana y el ADN de esta cepa bacteriana.

Tagetes lucida Cav. (Asteraceae=Compositae) is used for treating stomach infections. The study focused on evaluating the composition and antimicrobial effect of an extract of T. lucida Cav. The plant extracted with ethanol at 10% w/v, and the extract composition analyzed by Rp-HPLC-MS. The antibacterial effect was evaluated against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Salmonella choleraesuis using disk diffusion, microdilution and bioautography methods. The sytox and comet assays were used to evaluate the mechanism of action. In this way, nine phenolic compounds were detected in the extract of T. lucida. The extract exhibited activity only on S. aureus (MIC of 4.000 mg/ml). The bioautography revealed that the phenolic compounds could act synergistically. The sytox and comet tests showed an antibacterial action of the extract on the bacterial membrane and DNA of this bacterial strain.

Actividad antioxidante y marcha fitoquímica de los capítulos de Tagetes filifolia Lag. "pacha anís" / Antioxidant activity and phytochemical screening of capitula of Tagetes filifolia Lag. or "pacha anís"

Objetivo: Estudiar la actividad antioxidante y marcha fitoquímica de los capítulos de Tagetes filifolia Lag. "pacha anís".Materiales y métodos: Estudio de tipo experimental en el cual se empleó 5 kg de los capítulos de la planta medicinal Tagetes filifolia lag., provenientes de Junín. Se usó el método de cribado fitoquímico de Olga Lock para la marcha fitoquímica y el método DPPH para la determinación de la actividad antioxidante. Se dividió la muestra en 3 grupos: etéreo, alcohol etílico y agua destilada a concentraciones de 100, 50 y 5 µg/ml.Resultados: Se encontró fenoles en cantidades abundantes tanto en el extracto en agua destilada como en el extracto en alcohol etílico, además este último tuvo cantidades moderadas de quinonas. Por otro lado, el extracto en alcohol etílico fue el que presentó el mayor porcentaje de captación de radicales libres (91.26%) a una concentración de 100 µg/ml, similares resultados se encontró con el extracto etéreo (88.94%) y el extracto en agua destilada (75.58%).Conclusiones: Los principales componentes químicos fueron fenoles y quinonas. El mayor efecto antioxidante se obtuvo del extracto etanólico de la planta Tagetes filifolia a una concentración de 100 µg/ml.

Biofilm inhibition activity of traditional medicinal plants from Northwestern Argentina against native pathogen and environmental microorganisms

Abstract: INTRODUCTION: Plants have been commonly used in popular medicine of most cultures for the treatment of disease. The in vitro antimicrobial activity of certain Argentine plants used in traditional medicine has been reported. The aim of this study was to investigate the antimicrobial, anti-biofilm, and anti-cell adherence activities of native plants (Larrea divaricata, Tagetes minuta, Tessaria absinthioides, Lycium chilense, and Schinus fasciculatus) collected in northwestern Argentina. METHODS: The activities of the five plant species were evaluated in Bacillus strains and clinical strains of coagulase-negative Staphylococcus isolated from northwestern Argentina and identified by 16S rDNA. RESULT: Lycium chilense and Schinus fasciculatus were the most effective antimicrobial plant extracts (15.62µg/ml and 62.50µg/ml for Staphylococcus sp. Mcr1 and Bacillus sp. Mcn4, respectively). The highest (66%) anti-biofilm activity against Bacillus sp. Mcn4 was observed with T. absinthioides and L. divaricate extracts. The highest (68%) anti-biofilm activity against Staphylococcus sp. Mcr1 was observed with L. chilense extract. T. minuta, T. absinthioides, and L. divaricata showed percentages of anti-biofilm activity of between 55% and 62%. The anti-adherence effects of T. minuta and L. chilense observed in Bacillus sp. Mcn4 reflected a difference of only 22% and 10%, respectively, between anti-adherence and biofilm inhibition. Thus, the inhibition of biofilm could be related to cell adherence. In Staphylococcus sp. Mcr1, all plant extracts produced low anti-adherence percentages. CONCLUSION: These five species may represent a source of alternative drugs derived from plant extracts, based on ethnobotanical knowledge from northwest Argentina.

Determination of volatile organic compounds of Tagetes filifolia Lag. (Asteraceae) from Córdoba (Argentina) using HS-SPME analysis / Determinación de compuestos orgánicos volátiles de Tagetes filifolia Lag. (Asteraceae) proveniente de Córdoba (Argentina) utilizando análisis por HS-SPME

A headspace solid-phase microextraction (HS-SPME) method followed by gas chromatography-mass spectrometry (GC-MS) and gas chromatography-flame ionization detection (GC–FID) are described for the analysis of volatile compounds in Tagetes filifolia Lag. The composition of the total aerial parts of the plant (whole plant) and the inflorescences, leaves and stems were studied separately using HS-SPME. As a result, 54 compounds were determined, of which 47 were identified. The major components observed in this analysis were trans-anethole and estragole. The HS-SPME method used for the analysis of volatile compounds of T. filifolia is simple, fast, effective, free from the use of solvents, and permits by an analysis of small amounts of samples to achieve better results in terms of the determination of the composition than those reported in the literature for the analysis of essential oil.

Se realizó el análisis de los componentes volátiles de Tagetes filifolia Lag. utilizando el método de microextracción en fase sólida del espacio de cabeza con análisis posterior por cromatografía de gases acoplada a espectrometría de masas y por cromatografía de gases con detección por ionización de llama. Se estudio la composición de la planta entera así como también la de las inflorescencias, hojas y tallos por separado empleando el método de HS-SPME. Como resultado, se determinaron 54 compuestos de los cuales 47 fueron identificados. Los componentes mayoritarios observados en este análisis fueron: trans-anetol y estragol. El método de HS-SPME utilizado para el análisis de los compuestos volátiles de T. filifolia es simple, rápido, efectivo, libre de la utilización de solventes, y permitió mediante el análisis de pequeñas cantidades de muestra alcanzar mejores resultados en cuanto a la determinación de la composición, que los reportados en literatura para el análisis del aceite esencial.

Chemical identification of Tagetes minuta Linnaeus (Asteraceae) essential oil and its acaricidal effect on ticks / Caracterização química e efeito acaricida do óleo essencial de Tagetes minuta Linnaeus (Asteraceae) em carrapatos

The control of tick species that affect animal production is vital for the economic welfare of the cattle industry. This study focused on testing the acaricidal activity of the essential oil from the leaves and stems of Tagetes minuta against several Brazilian tick species, including Rhipicephalus (Boophilus) microplus, Rhipicephalus sanguineus, Amblyomma cajennense and Argas miniatus. The chemical composition of the essential oil was determined by chromatography and spectroscopy analyses, which revealed the presence of monoterpenes. The adult immersion test (AIT) and the larval packet test (LPT) were used to evaluate the efficacy of T. minuta essential oil in tick management at concentrations of 2.5, 5, 10, 20 and 40%. The results demonstrated that the T. minuta essential oil had over 95% efficacy against four species of ticks at a concentration of 20%. These results suggest that the essential oil of T. minuta could be used as an environmentally friendly acaricide.(AU)

O controle de carrapatos que causa impacto na produção de bovinos possui importância econômica para a cadeia produtiva. Neste trabalho objetivou-se testar a atividade acaricida do óleo essencial das folhas e caules de Tagetes minuta contra várias espécies de carrapatos brasileiros, incluindo Rhipicephalus (Boophilus) microplus, Rhipicephalus sanguineus, Amblyomma cajennense e Argas miniatus. A composição química do óleo foi determinada por GC-MS e análises de espectroscopia de RMN, que revelaram a presença de monoterpenos. Na avaliação destas substâncias no controle do carrapato foram empregados os testes de imersão de adulto (TIA) e o de pacote de larvas (TPL) para o extrato de óleo de T. minuta nas concentrações de 2,5%; 5%; 10%; 20% e 40%. Os resultados do TPL e TIA demonstraram que o óleo essencial na concentração de 20% de T. minuta apresenta eficácia superior a 95% nas quatro espécies de carrapato. Estes resultados sugerem que o óleo essencial de T. minuta pode ser usado como um acaricida eficaz e com baixo impacto ambiental.(AU)

Determination of volatile organic compounds of tagetes Argentina Cabrera (Asteraceae) using HS-SPME analysis / Determinación de compuestos orgánicos volátiles de tagetes Argentina Cabrera (Asteraceae) utilizando análisis por HS-SPME

A headspace solid-phase microextraction (HS-SPME) method followed by gas chromatography-mass spectrometry (GC-MS) and gas chromatography-flame ionization detection (GC-FID) is described for the analysis of volatile compounds in Tagetes argentina Cabrera. The composition of the totality of the aerial parts of the plant (whole plant) and the inflorescences, leaves and stems was studied separately using HS-SPME. As a result, 53 compounds were determined, of which 39 were identified. The major components were: cis-tagetenone, trans-tagetenone, trans-tagetone and cis-tagetone.Using a much smaller amount of samples, a shorter extraction time and a very simple procedure, the HS-SPME method can achieve similar or better results than those obtained by EO analysis. In conclusion, the HS-SPME method is simple, rapid, effective and free of solvent, and can be used for the analysis of volatile compounds in samples of different populations of T. Argentina.

Se realizó el análisis de los componentes volátiles de T. argentina Cabrera utilizando el método de micro-extracción en fase sólida del espacio de cabeza con análisis posterior por cromatografía de gases acoplada a espectrometría de masas y por cromatografía de gases con detección por ionización de llama. Se estudio la composición de la totalidad de las partes aéreas de la planta (planta entera) como así también de las inflorescencias, hojas y tallos por separado empleando el método de HS-SPME. Como resultado, se determinaron 53 compuestos de los cuales 39 fueron identificados. Los componentes mayoritarios fueron: cis-tagetenona, trans-tagetenona, trans-tagetona and cis-tagetona. Utilizando una muy pequeña cantidad de muestra, un corto periodo de tiempo y un procedimiento muy simple se lograron similares o mejores resultados a aquellos reportados mediante el análisis del aceite esencial. En conclusión, el método de HS-SPME es simple, rápido, efectivo, libre de la utilización de solventes y puede ser fácilmente implementado para el análisis de componentes volátiles provenientes de muestras de diferentes poblaciones de T. Argentina.

Efficacy of indigenous plant extracts on the malaria vector Anopheles subpictus Grassi (Diptera: Culicidae).

Background & objectives: Mosquito control is facing a threat due to the emergence of resistance to synthetic insecticides. Insecticides of plant origin may serve as suitable alternative biocontrol techniques in the future. The purpose of the present study was to assess the ethyl acetate, acetone and methanol extracts of Andrographis paniculata, Eclipta prostrata and Tagetes erecta leaves tested for oviposition-deterrent, ovicidal and repellent activities against malaria vector, Anopheles subpictus Grassi (Diptera: Culicidae). Methods: The dried leaves of the three plants were powdered mechanically and extracted with ethyl acetate, acetone and methanol. One gram of crude extract was first dissolved in 100 ml of acetone (stock solution). From the stock solution, test solution concentrations of 31.21- 499.42 mg/l for oviposition- deterrence assay and repellency and 15.60 - 998.85 mg/l were used in ovicidal assay. The percentage oviposition- deterrence, hatching rate of eggs and protection time were calculated. One-way analysis of variance was used for the multiple concentration tests and for per cent mortality to determine significant treatment differences. Results: The percentage of effective oviposition repellency was highest at 499.42 mg/l and the lowest at 31.21 mg/l in ethyl acetate, acetone and methanol extracts of A. paniculata, E. prostrata and T. erecta. The oviposition activity index (OAI) value of ethyl acetate, acetone and methanol extracts of A. paniculata, E. prostrata and T. erecta at 499.42 mg/l were -0.91, -0.93, -0.84, -0.84, -0.87, -0.82, -0.87, -0.89 and -0.87, respectively. Mortality (no egg hatchability) was 100 per cent with ethyl acetate and methanol extracts of A. paniculata, E. prostrata and T. erecta at 998.85 mg/l. The maximum adult repellent activity was observed at 499.42 mg/l in ethyl acetate extracts of A. paniculata, E. prostrata and methanol extracts of T. erecta, and the mean complete protection time ranged from 120 to 150 min with the different extracts tested. Interpretation & conclusions: The acetone extract of A. paniculata, methanol extract of E. prostrata and T. erecta showed good oviposition-deterrent, ovicidal and repellent activities respectively. These results suggest that the leaf extracts of A. paniculata, E. prostrata and T. erecta may have the potential to be used as an ideal eco-friendly approach for the control of the An. subpictus.

Antioxidant activity of Blechnum chilense (Kaulf.) Mett., Curcuma domestica Valeton and Tagetes verticillata Lag. & Rodriguez / Actividad antioxidante de Blechnum chilense (Kaulf.) Mett., Curcuma domestica Valeton y Tagetes verticillata Lag. & Rodríguez

Because of the increasing interest in improving human health worldwide, phytochemical antioxidants from medicinal and food plants are of great interest. The search for new sources of antioxidants is important for the best use of biodiversity. The objective of this work was to evaluate the antioxidant activity and the total phenolic compounds with DPPH and Folin-Ciocalteu assays for extracts and fractions of Blechnum chilense, Curcuma domestica and Tagetes verticillata. B. chilense water-methanolic and EtOAc fractions, follows of C. domestica EtOAc extract showed an important quantity of total phenolic compounds. Compared with Aristotelia chilensis MeOH extract, T. verticillata extract showed good activity, follows by EtOAc fraction from B. chilense and by EtOAc extract from C. domestica, with very similar results with n-hexane fraction from B. chilense and petroleum ether extract from C. domestica. All of these results were greater than alpha-tocopherol DPPH scavenging activity. The results suggest that all plants studied could be are new sources of antioxidants and the work are following with the identification of these compounds.

Debido al creciente interés mundial en el mejoramiento de la salud humana los antioxidantes provenientes de plantas medicinales y alimenticias se han convertido en compuestos de gran interés. La búsqueda de nuevas fuentes de antioxidantes es importante para el mejor uso de la biodiversidad. El objetivo de este trabajo fue evaluar la actividad antioxidante y el contenido de fenoles totales usando el método de Folin-Ciocalteu y la actividad inhibitoria del radical DPPH de fracciones y extractos de Blechnum chilense (Kaulf.) Mett, Curcuma domestica Valeton y Tagetes verticillata Lag. & Rodr. Las fracciones acuosa-metanólica y EtOAc de B. chilense, seguida del extracto EtOAc de C. domestica, mostraron una cantidad importante de compuestos fenólicos. La prueba con DPPH mostró que la actividad secuestrante más importante, comparada con la del extracto metanólico de Aristotelia chilensis (Molina) Stuntz, fue la del extracto de T. verticillata, seguido de la fracción EtOAc de B. chilense y el extracto EtOAc de C. domestica, con resultados similares a la fracción hexánica de B. chilense y el extracto obtenido con éter de petróleo de C. domestica, superando todas la actividad secuestrante de DPPH del alfa-tocoferol. Los resultados sugieren que todas las plantas estudiadas podrían ser nuevas fuentes de antioxidantes y se está trabajando para la identificación de los compuestos responsables de la actividad.

Phytochemistry of Tagetes minuta L. (Asteraceae) from Córdoba, Argentina: comparative study between essential oil and HS-SPME analyses / Fitoquímica de Tagetes minuta L. (Asteraceae) originario de Córdoba, Argentina: estudio comparativo entre análisis del aceite esencial y análisis por HS-SPME

A headspace solid-phase microextraction (HS-SPME) method followed by gas chromatography-mass spectrometry (GC-MS) and gas chromatography-flame ionization detection (GC–FID) is described for the analysis of volatile compounds in Tagetes minuta L. Five types of SPME commercial fibers including PA, PDMS, CAR-PDMS, PDMS–DVB and DVB–CAR–PDMS were investigated and the best extraction was achieved with the mixed fiber DVB–CAR–PDMS. Parameters for HS-SPME in terms of equilibrium time of HS, fiber exposition time and extraction temperature were also investigated. Additionally, the composition of inflorescences, leaves and stems was also studied separately by HS-SPME. As a result, 68 compounds were determined and 53 were identified. A comparison was made between results obtained by HS-SPME–GC–MS and steam distillation of essential oil of the aerial parts of the plant. In both analyses, the major components were: cis-tagetenone and trans-tagetenone. Using much smaller samples, a shorter extraction time and a simpler procedure, the HS-SPME method can achieve similar results to those obtained by EO analysis. In conclusion, the HS-SPME method is simple, rapid, effective and free of solvent, and can be used for the analysis of volatile compounds in samples of different populations of T. minuta.

Se realizó el análisis de los componentes volátiles de T. minuta L. utilizando el método de micro-extracción en fase sólida del espacio de cabeza con análisis posterior por cromatografía de gases acoplada a espectrometría de masas y por cromatografía de gases con detección por ionización de llama. Se estudiaron cinco tipos de fibras comerciales que incluyeron a PA, PDMS, CAR-PDMS, PDMS–DVB y DVB–CAR–PDMS y se estableció que la fibra de DVB–CAR–PDMS es la que posee mejor comportamiento en el proceso de extracción. Se determinó el efecto del tiempo de equilibrio del espacio de cabeza, de la temperatura de extracción y del tiempo de exposición de la fibra sobre el proceso de HS-SPME. Adicionalmente, también se estudio por separado la composición de inflorescencias, hojas y tallos empleando el método de HS-SPME. Como resultado de este estudio se determinaron 68 componentes de los cuales 53 fueron identificados. Por otra parte se realizó una comparación de los resultados HS-SPME con el análisis del aceite esencial obtenido de las partes aéreas de la planta. En ambos casos, los componentes mayoritarios fueron: cis-tagetenona y trans-tagetenona. Utilizando una muy pequeña cantidad de muestra, un corto periodo de tiempo y un procedimiento más simple se lograron similares resultados a los obtenidos mediante el análisis del aceite esencial. En conclusión, el método de HS-SPME desarrollado es simple, rápido, efectivo y libre de la utilización de solventes, puede ser fácilmente implementado para el análisis de componentes volátiles provenientes de muestras de diferentes poblaciones de T. minuta.

Estabelecimento de metodologia para alimentação de Aedes aegypti (Diptera-Culicidae) em camundongos swiss e avaliação da toxicidade e do efeito residual do óleo essencial de Tagetes minuta L (Asteraceae) em populações de Aedes aegypti / Establishment of the feeding methodology of Aedes aegypti (Diptera-Culicidae) in Swiss mice and evaluation of the toxicity and residual effect of essential oil from Tagetes minuta L (Asteraceae), in populations of Aedes aegypti

Objetivou-se desenvolver um procedimento de alimentação de fêmeas de Aedes aegypti que não cause estresse em camundongo swiss e avaliar a toxicidade e o efeito residual do óleo essencial de Tagetes minuta L (Asteraceae) em populações de Aedes aegypti. Camundongos anestesiados: um observado tempo de sedação e outro colocado em gaiola para alimentação de fêmeas. Óleo essencial, diluído em acetona, foi utilizado em bioensaios para avaliação das concentrações letais em larvas de Bauru, SP e São José do Rio Preto, SP, respectivamente, sensíveis e resistentes ao temephos. Os dados obtidos foram comparados com a cepa Rockefeller-EUA. O procedimento com camundongos foi aprovado. Não houve diferença entre as populações quanto à susceptibilidade a Tagetes minuta e os ensaios demonstraram CL50 de 0,24, 0,25 e 0,21mL L-1 e CL99,9 em 0,35, 0,39 e 0,42mL L-1, respectivamente, para Rockfeller, Bauru e São José do Rio Preto. Não foi observado efeito residual da solução.

The objectives here were to develop a procedure for feeding females of Aedes aegypti that does not cause stress in Swiss mice and to evaluate the toxicity and residual effect of essential oil from Tagetes minuta L. (Asteraceae) in Aedes aegypti populations. Two mice were anesthetized: one was used to observe the duration of sedation and the other was placed in a cage to feed the female mosquitoes. Essential oil was diluted in acetone and used in bioassays to assess the lethal concentrations in larvae from the Cities of Bauru (SP) and São José do Rio Preto (SP) that were sensitive and resistant to temephos, respectively. The data obtained were compared with the American Rockefeller strain. The procedure with mice was approved. There was no difference between the populations regarding susceptibility to Tagetes minuta, and the assays showed LC50 of 0.24, 0.25 and 0.21 ml/l and LC99.9 of 0.35, 0.39 and 0.42 ml/l, for Rockefeller, Bauru and São José do Rio Preto, respectively. The solution did not show any residual effect.

A source of almost pure methyl chavicol: volatile oil from the aerial parts of Tagetes lucida (Asteraceae) cultivated in Costa Rica

The plant Tagetes lucida Cav.(syn.T.florida Sweet,T.schiedeana Less.)is an aromatic herb distributed naturally from Mexico to Honduras,at elevations between 1 000 and 2 000 m.It is used as a spice,for medicine,as insecticide and as ornamental plant.It is cultivated commercially in Costa Rica as a spice herb;it contains an oil having an anise-like odor,and the fresh aerial parts of this plant are sold in the supermarket as a substitute of tarragon (Artemisia dracunculus L.).The essential oils isolated from aerial parts bought,at May and October,in a supermarket in San José (Costa Rica).Fresh flowering aerial parts,flowers and leaves plus stems, were subjected to hydrodistillation for 3 hr using a modified Clevenger-type apparatus.The distilled oils were collected and dried over anhydrous sodium sulphate and stored in a freezer (0-10 °C).The light yellow green oil yield was about 0.07%(v/w).GC/MS analyses were performed using a Shimadzu GCMS-QP5050 apparatus and CLASS 5000 software with Wiley 139 computer database.Identification of the components of the oil was performed using the retention indices,which were calculated in relation to a homologous series of hydrocarbons, and by comparison of their mass spectra with those published in the literature or those of our own database. Thirty compounds were identified,of which methyl chavicol (95-97%)was the major constituent.From flower oil,two bithienyls were detected as minor constituents

Los aceites esenciales extraídos de las partes aéreas de la planta Tagetes lucida Cav.cultivada en Costa Rica y utilizada como condimento,fue estudiado mediante la técnica de GC/MS en combinación con los índices de retención.Se identificaron treinta compuestos.El componente mayoritario resultó ser metil chavicol (estragol)en un 95-97%.En el aceite de las flores se detectaron e identificaron dos compuestos minoritarios que resultaron ser bitienilos no informados anteriormente como constituyentes de esta planta