2010 The Authors 2010 Boletn Latinoamericano y del Caribe de Plantas Medicinales y Aromticas, 9 (2), 123-126 BLACPMA ISSN

N 0717 7917 Artculo Original | Original Article

Aromatic plants from Patagonia: chemical composition and antimicrobial activity of the essential oil of Senecio mustersii and S.subpanduratus
[Plantas aromticas de la Patagonia: Composicin qumica y actividad antimicrobiana del aceite esencial de Senecio mustersii y S.subpanduratus]
Luz ARANCIBIA 1, Cecilia NASPI 1, Graciela PUCCI 2, Mara ARCE 3
1

Ctedra de Qumica Orgnica, 2Ctedra de Microbiologa ,3Ctedra de Botnica, Facultad de Ciencias Naturales, Universidad Nacional de la Patagonia S J B, Comodoro Rivadavia, Chubut, Argentina.

Abstract
The objective of this investigation was the determination of the antimicrobial activity of two plants from Patagonia Argentina; Senecio mustersii and Senecio subpanduratus (Asteraceae). Until the present day, no previous studies have been reported on the composition of the essential oil for these two species of Senecio. The essential oils were obtained by hydrodistillation with a yield of 0.81% for Senecio subpanduratus and 0.71% for Senecio mustersii, expressed as ml of essential oil per 100 g of fresh vegetable matter. The activity against bacteria and yeast was tested; Senecio mustersii showed activity against S.aureus and Senecio subpanduratus against all tested bacteria (S.aureus, E.coli and P. aeruginosa). Senecio mustersii didnt showed antifungal activity; meanwhile Senecio subpanduratus was active against some species of Candida. Keywords: Essential Oils; Senecio; Antifungal activity; Antibacterial activity.

Resumen
El objetivo de la investigacin fue la determinacin de la actividad antimicrobiana de los aceites esenciales de dos especies del gnero Senecio (Asteraceae) de la regin Patagnica: Senecio mustersii y S. subpanduratus. Hasta el momento, no se han reportado estudios sobre la composicin del aceite esencial para estas dos especies de Senecio. Los aceites esenciales fueron obtenidos mediante hidrodestilacin logrndose un rendimiento de 0.81 % para Senecio subpanduratus y de 0.72% para Senecio mustersii, expresado como ml de aceite esencial por cada 100 g de material vegetal fresco. Se evalu la actividad frente a bacterias y levaduras de importancia clnica: Senecio mustersii presenta actividad antibacteriana frente a S. aureus y Senecio subpanduratus para todas las bacterias testeadas (S.aureus, E.coli y P. aeruginosa). Senecio mustersii no present actividad antifngica, mientras que Senecio subpanduratus actividad contra algunas especies de Candida. Palabras Clave: Aceites Esenciales; Senecio; Actividad Antifngica; Actividad Antibacteriana. List of Abbreviations: HRP (Herbario Regional Patagnico); CG-FID-MS (Gas chromatography-Flame ionization detector-Mass spectrum); ATCC (American type culture collection); NIM (Nmero Instituto Malbrn);ANLIS (Administracin nacional de laboratorios e institutos de salud); MIC (minimal inhibition concentration);
Recibido | Received: December 18, 2009. Aceptado en Versin Corregida | Accepted in Corrected Version: February 7, 2010. Publicado en Lnea | Published Online March 25, 2010 Declaracin de intereses | Declaration of interests: authors have no competing interests. Financiacin | Funding: Universidad Nacional de la Patagonia San Juan Bosco This article must be cited as: Luz Arancibia, Cecilia Naspi, Graciela Pucci, Mara Arce. 2010. Aromatic plants from Patagonia: chemical composition and antimicrobial activity of the essential oil of Senecio mustersii and S.subpanduratus. Bol Latinoam Caribe Plant Med Aromat 9(2):123 126 . EPub March . 2010}.

*Contactos | Contacts: luz@unpata.edu.ar ; cecinaspi@yahoo.com.ar

BLACPMA es una publicacin de la Cooperacin Latinoamericana y Caribea de Plantas Medicinales y Aromticas This is an open access article distributed under the terms of a Creative Commons Attribution-Non-Commercial-No Derivative Works 3.0 Unported Licence. (http://creativecommons.org/licenses/by-nc-nd/3.0/ ) which permits to copy, distribute and transmit the work, provided the original work is properly cited. You may not use this work for commercial purposes. You may not alter, transform, or build upon this work. Any of these conditions can be waived if you get permission from the copyright holder. Nothing in this license impairs or restricts the author's moral rights. Este es un articulo de Acceso Libre bajo los trminos de una licencia Atribucin Creativa Comn -No Comercial-No trabajos derivados 3.0 Internacional (http://creativecommons.org/licenses/by-ncnd/3.0/deed.es) Usted es libre de copiar, distribuir y comunicar pblicamente la obra bajo las condiciones siguientes: Reconocimiento. Debe reconocer los crditos de la obra de la manera especificada por el autor o el licenciador (pero no de una manera que sugiera que tiene su apoyo o apoyan el uso que hace de su obra). No comercial. No puede utilizar esta obra para fines comerciales. Sin obras derivadas. No se puede alterar, transformar o generar una obra derivada a partir de esta obra. Al reutilizar o distribuir la obra, tiene que dejar bien claro los trminos de la licencia de esta obra. Alguna de estas condiciones puede no aplicarse si se obtiene el permiso del titular de los derechos de autor. Nada en esta licencia menoscaba o restringe los derechos morales del autor.

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Antimicrobial activity of Senecio mustersii and S.subpanduratus

INTRODUCTION There are about 3000 species of Senecio around the world, mainly in hilly areas. In Argentina there are more than 270 species, most of them in the Andes range and in Patagonia. (Cabrera, 1971) Senecio mustersii and S.subpanduratus (Asteraceae) are native species that grow spontaneously near Comodoro Rivadavia, Chubut. Senecio subpanduratus grows in Chubut and Tierra del Fuego meanwhile Senecio mustersii Speg. Var. mustersii can be found in Ro Negro, southwest of Chubut and east of Santa Cruz (Cabrera, 1971). This genus contains essential oils on aerial parts of the plant and also sesquiterpenes in particular furanoeremophilanes (Bohlmann et al., 1986, Salmeron et al., 1983; Torres et al., 1999) and pyrrolizidine alkaloids. The objective of this work is to study in vitro antifungal and antibacterial activity for these species of Senecio. Essential oils from many aromatic plants, included Senecio have been studied because of their chemical composition and antimicrobial activity (Perez et al 1999, El-Shazly 2002). Thus Schinus polygamus (Gonzalez et al.) and S. johnstonii (Malizia et al.) from Patagonia is reported on the composition and antimicrobial activity of the essential oil. MATERIALS AND METHODS Plant Material The plant material was obtained from plants at 3 Km north from Comodoro Rivadavia city, province of Chubut (Argentina), during May of 2006. The species were identified by botanist (M.E.Arce) and kept in the Patagonia Regional Herbarium (Universidad Nacional de la Patagonia San Juan Bosco) under the following herbarium numbers: Senecio subpanduratus O. Hoffm. HRP 6867 and Senecio mustersii Speg. Var. Mustersii HRP 6866. Essential oil Extraction Fresh aerial parts of Senecio mustersii and S.subpanduratus were cut into small pieces. The essential oils were obtained by hydrodistillation during 4 hours in a Clevenger-type apparatus. Gas Chromatography-Mass Spectrometry Analyses were performed by CG-FID-MS in a Perkin Elmer Clarus 500 provided with a unique
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injector Split type (1:100 Relation) and with two fused silica capillary columns: polyethylenglycol and 5% phenyl-95% methyl silicone, (both 60 m x 0.25 mm x 25m df). Polar column is coupled to a FID meanwhile non polar column to a FID-mass detector (70 eV) through an MSVent system. The carrier gas was Helium (flow rate: 1.87 ml/min). Column temperature was initially 90C, and then increased to 225C at 3/min rate (15 min). Samples were diluted (10% v/v in ethanol) and 0.2 l were injected. The constituents of the essential oils were identified on the basis of their GC retention indices (RI) with reference to a homologous series of nalkanes (C8-C20) and by comparison of their mass spectrum with reported data (Adams, Wiley, and Nist/EPA/NIH Mass Spectral Library). Antibacterial and Antifungal assays Antimicrobial and antifungal activity was assayed against eight microorganisms including Gram (+) and (-) bacteria and yeast: Staphylococcus aureus (ATCC 29213), Pseudomona aeruginosa (ATCC 27853), Escherechia coli (ATCC 25299), Candida albicans (NIM 982879), Candida tropicalis (ATCC 2000956), Candida parapsilopsis (ATCC 22019), Candida krusei (ATCC 6258), Candida guillermondi and Candida glabrata donated by ANLIS Malbrn Institute. The antimicrobial activity was performed in solid phase by Agar Dilution Method (Wright et al, 1983; Ruhnke et al, 1996). The oil and its dilutions (1/125, 1/250, 1/500, 1/1000 y 1/2000 v/v) were initially mixed with sterile nutritive agar for bacteria and 4% glucose-sabouread for yeast and then stirred for a minute in a vortex at 3000 rpm. An inoculum of 106 cells per ml was mixed with the medium, bacteria and yeast culture were incubated for 24 hours at 37C and 48 hours at 28C respectively. The MIC endpoint was determined visually by recording the lowest concentration of the essential oil that prevented the appearance of visible growth. RESULTS AND DISCUSSION Essential oil Extraction The amounts of essential oils obtained in both Senecio species were similar. The yield was 0.81% for Senecio subpanduratus and 0.71% for Senecio mustersii, expressed as ml of essential oil per 100 g of fresh vegetable matter.

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Antimicrobial activity of Senecio mustersii and S.subpanduratus

Chromatography-Mass Spectrometry A quantitative and qualitative variation between both Senecio species was apparent. Relative percentages of the oil constituents are showed in table 1, listed in order of elution from the used column. Twenty four compounds were identified for S.mustersii (95.2%) and twenty one for S.subpanduratus (92.9%).
Table 1: Chemical composition of essential oil of S.mustersii and S.subpanduratus (expressed as percentages). Compounds tuyene -pinene sabinene myrcene pinene 3-carene p-cymene limonene perillene -pinene oxide -canfolenal pinocarveol trans+sabinol trans sabina ketone pinocarvone Terpinen-4-ol p-cimen-8-ol -terpineol myrtenol + myrtenal myrtenal pinocarveol acetate trans pinocarveol acetate cis kessane spathulenol -oplopenone Tcadinol Epi--murolol -cadinol TOTAL S. mustersii S. subpanduratus 0.1 53.3 1.6 2.3 21.2 0.2 1.7 1.8 tr 1 0.3 2.1 0.4 0.5 tr 0.4 1.3 0.9 1.1 0.9 0.2 1.2 0.2 0.8 1.7 95.2 1.7 22.1 23.8 2.6 11.9 0.4 8.7 1.2 0.2 0.6 0.4 10.2 0.4 0.7 0.7 0.3 2.9 0.4 2.5 tr 0.2 1 92.9

The essential oil of S. mustersii was characterized by -pinene (53.3%) and -pinene (21.2%) as major components. Meanwhile in S.subpanduratus -pinene (22.1%) and -pinene (11.9%) were also detected, in addition to an important amount of sabinene (23.8%), terpinen-4-ol (10.2%) and p-cymene (8.7%). Antibacterial and Antifungal assays The results are showed in table 2, where the antimicrobial activity was determined by the appearance of visible growth. The isolated essential oil of S. subpanduratus showed antimicrobial activity against the three bacterial strains tested and also against C.albicans, C. parapsilisis and C.guillermondii, a fact that may become relevant, given the pathogenic properties of these strains. The isolated essential oil S mustersii had only antimicrobial activity against S.aureus. This difference can be explained by the chemical composition of the essential oils.
Table 2: Antimicrobial activity of essential oil of S.mustersii and S.subpanduratus Essential oil Dilutions (v/v) Senecio mustersii 1/1000 1/2000 1/125 1/250 1/500 Bacteria S. aureus E. coli A P. aeruginosa Yeast C. albicans C. tropicalis C. parapsilosis C. guillermondii C. krusei C. glabrata + + + + + + + + + + + + Senecio subpanduratus 1/1000 1/2000 1/125 1/250 1/500

+ -

+ -

+ + +

+ + +

(+)Antimicrobial activity. (-) No antimicrobial activity.

Antimicrobial activity of essential oils is difficult to attribute to a specific compound, probably because of the complexity of its composition and also for the synergic effects that may exist between the major components. Despite this, there are studies that explain the antimicrobial activity of some components of the essential oil. It has been demonstrated that and pinene are able not only to destroy cellular integrity, but also inhibit respiration and ion transport processes. They also increase the membrane permeability in yeast

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Antimicrobial activity of Senecio mustersii and S.subpanduratus

cells (Magwa et al 2006, Andrews 1980, Uribe 1985).There are other studies of essential oils that demonstrate effects also on gram negative membrane (Helander, 1998). CONCLUSIONS This is the first report about chemical composition of essential oil and biological activity of these species of Senecio from the East Central Area of Patagonia. The high percentage of pinenes ( and ) in both species could play an important role in defensive mechanism and adaptation to dessert area. The differences in antimicrobial activity of both species of Senecio could be related to oxygenated derivates, which are in higher percentage in S.subpanduratus. ACKNOWLEDGES We would like to thank ANLIS (Administracin Nacional de Laboratorios e Institutos de Salud) Dr. Carlos Malbrn, for donating the strains that were used in this investigation. REFERENCES
Andrews RE, Parks LW, Spence KD. 1980. Some effects of Douglas fir terpenes on certain microorganism. Appl. Environ. Microbiol. 40: 301-304. Bohlmann F, Zdero C, Jakupovic J, Grenz M, Castro V, King RM, Robinson H, Vincent LPD. 1986. Further Pyrrolizidine Alkaloids and Furanoeremophilanes from Senecio species. Phytochemistry. 25(5): 1151-1159. Cabrera A. 1971. Compositae, pp. 242-243. In Correa MN: Flora Patagnica, parte 7. Coleccin Cientfica del INTA. Buenos Aires. Argentina De Salmeron MSA, Kavka J, Giordano OS. 1983. Furanoeremophilanes in Senecio filaginoides and S.pinnatus. Planta Med.. 47: 221-223. El-Shazly A, Doral G, Wink M. 2002. Chemical Composition and Biological Activity of the Essential Oils of Senecio aegytius var.discoides Boiss. Z. Naturforsch. 57 (c): 434-439.

Gonzlez S, Guerra P, Bottaro H, Demo M, Zunino M, Zygadlo J. 2004. Aromatics plants from Patagonia, Antimicrobial activity and chemical composition of Schinus polygamus (Cav.) Cabrera essential oil. Flavour Fragrance J. 19 (1): 36-39. Helander IM, Alakomi HL, Kyosti LK, Mattiala-sndholm Y, Pol I, Smid EJ, Gorris GM, von Wright A. 1998. Characterization of the action of selected essential oil components on Gram-negative bacteria. J. Agric. Food Chem. 46: 3590-3595. Kamatou GPP, Viljoen AM, Figueiredo AC, Tilney PM, Van Zyl RL, Barroso JG, Pedro LG, Van Vuuren, SF. 2007. Trichomes, essential oil composition and biological activities of Salvia albicaulis Benth. and S.dolomitica Codd, two species from the Cape region of South Africa. S. Afr. J. Bot. 73: 102-108. Magwa ML, Gundidza M, Gweru N, Humphrey G. 2006. Chemical composition and biological activities of essential oil from the leaves of Sesuvium portulacastrum. J Ethnopharmacol. 103: 85-89. Malizia RA, Molli JS, Cardell DA, Retamar JA; Arancibia LA; Arce ME. 2006. Aceite Esencial de Schinus johnstonii Barkley. Naturalia Patagnica. 3 (1):45-48. Perez C, Agnese AM, Cabrera JL. 1999. The essential oil of Senecio graveolens (Compositae): Chemical composition and antimicrobial activity test. J Ethnopharmacol 66: 91-96. Ruhnke M, Schmidt-Westhausen A, Engelmann E, Trautmann M. 1996. Comparative evaluation of three antifungal susceptibility test methods for Candida albicans isolates and correlation with response to fluconazole therapy. J Clin Microbiol. 34(12): 3208 3211. Torres P, Ayala J, Grande C, Anaya J, Grande M. 1999. Furanoeremophilane derivates from Senecio flavus. Phytochemistry. 52: 1507-1513. Uribe S, Ramirez T, Pena A. 1985. Effects of -pinene on yeast membrane functions. J. Bacteriol. 161: 195-200. Wright LR, Scott EM, Gorman SP. 1983.The sensitivity of mycelium, arthrospores and microconidia of Trichophyton mentagrophytes to imidazoles determined by in-vitro tests. J. Antimicrob. Chemother. 12: 317327.

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