Núm. 31, pp. 21-29, ISSN 1405-2768; México, 2011 MORPHOANATOMICAL CHARACTERIZATION AND ANTIMICROBIAL ACTIVITY OF TILLANDSIA IMPERIALIS (BROMELIACEAE) J. Alejandro Vite-Posadas1, Alicia E. Brechú-Franco1, Guillermo Laguna-Hernández1, M. Gabriela Rojas-Bribiesca2 and H. Reyna Osuna-Fernández1. 1Laboratorio de Estructura y Fisiología de Plantas, Facultad de Ciencias, Universidad Nacional Autónoma de México, Ciudad de México; 2Laboratorio de Microbiología, Centro de Investigación Biomédica del Sur (CIBIS), IMSS, Xochitepec, Morelos, México. ABSTRACT trichomes, tetracyclic stomata and non- sclerotic hypodermis with spherical silica Tillandsia imperialis C.J. Morren ex Roezl is bodies. Microbiological research revealed a bromeliad used in Mexican folk medicine specifi c antimicrobial activity against re- mixed with other plant and animal parts as a spiratory pathogenic microorganisms such good remedy for respiratory diseases, which as Staphylococcus aureus and Streptococ- are one of the main causes of morbility and cus pyogenes as well as six other bacteria. mortality in the rural population of Mexico. These results indicate the existence of anti- The commercialization of medicinal plants microbial compounds in the extracts and a for therapeutic use must comply with high correlation between the traditional uses of standards for quality, safety and effi cacy; this plant and the experimental data. structural, microbiological and pharmaceuti- cal studies make a decisive contribution to Key words: antimicrobial activity, me- this quality control. The aim of this study dicinal plants, structural and histochemical was to analyze the morpho-anatomical char- characteristics, Tillandsia imperialis. acteristics and histochemistry of bracts and leaves of Tillandsia imperialis and evaluate RESUMEN their antimicrobial activity to corroborate the effectiveness of this plant for traditional Tillandsia imperialis (Bromeliaceae) es una uses with experimental data. Crude extracts planta utilizada en la medicina tradicional from the infl orescences and leaves were mexicana para tratar ciertas afecciones evaluated on Staphylococcus aureus (ATCC respiratorias, las cuales son una de las prin- 29213), Streptococcus pyogenes (ATCC cipales causas de morbilidad y mortalidad 08668), Escherichia coli (ATCC 25922), en la población de nuestro país. No obstante, Streptococcus faecalis (ATCC 29212), Sal- no se han realizado investigaciones que monella Typhi (ATCC 06539), Klebsiella corroboren su uso en la medicina tradi- pneumoniae (ATCC 13883) and Candida cional. Se realizó un estudio estructural e albicans (ATCC 10231). Results showed histoquímico de las brácteas y hojas de esta that T. imperialis possesses important mor- especie para aportar información útil para pho-anatomical characteristics useful for su correcta determinación farmacognóstica identifi cation, including mesic and peltate y se evaluó la actividad antimicrobiana 21 Núm. 31: 21-29 Marzo 2011 de los extractos crudos obtenidos de las in developing countries (Salud Pública infl orescencias y hojas para corroborar su Méx., 2004; WHO, 1998). Phytotherapy as uso en la medicina tradicional. La actividad a part of Therapeutics requires the rational antimicrobiana de los extractos crudos se development of herbal medicinal products evaluó sobre Staphylococcus aureus (ATCC with guaranteed quality, safety and effi cacy, 29213), Streptococcus pyogenes (ATCC along with accurate and assessable informa- 08668), Escherichia coli (ATCC 25922), tion as well as appropriate education in this Streptococcus faecalis (ATCC 29212), fi eld (Cañigueral, 2006). The confi rmation Salmonella Typhi (ATCC 06539), Klebsi- of the authenticity of the botanical samples ella pneumoniae (ATCC 13883) y Candida requires pharmacognostic studies. Few albicans (ATCC 10231). Las características scientifi c investigations have analyzed the morfoanatómicas útiles que se detectaron en leaf, stem or root anatomy, which is useful T. imperialis son tricomas mésicos, pelta- to guarantee the authenticity of medicinal dos, estomas tetracíclicos e hipodermis no botanical materials and in turn contributes to esclerótica con cuerpos esféricos de sílice. their identifi cation and characterization for El estudio antimicrobiano reveló actividad the quality control of the products derived contra microorganismos patógenos de vías from these plants (Arambarri & Mandrile, respiratorias como Staphylococcus aureus 1999). It is necessary to make botanical y Streptococcus pyogenes lo cual corrobora tests for plant materials from markets to y valida su uso en la medicina tradicional avoid the falsifi cation or substitution in the de México. use of any medicinal botanical material. Data acquired in scientifi c experimentation Palabras clave: actividad antimicrobiana, should guarantee and provide approaches estructura e histoquímica, planta medicinal, for quality control, as recommended by the Tillandsia imperialis. World Health Organization and the Mexican Health system. INTRODUCTION In spite of the diversity of medicinal herbs, For many people around the world, tra- the scientifi c knowledge available is very ditional medicine is still used and it is an limited as in the case of bromeliads. This important medicinal resource (McGaw et group exhibits great diversity of habitats al., 2000). Medicinal plants used in the in- and represents the third most important bo- digenous cultures from developing countries tanical family of Mexican monocotyledons are numerous and diverse (Nigenda et al., (Pulido-Esparza et al., 2004). Bromeliads 2001). The traditional medicine in Mexico have been used since ancient times and plays an important role in medical practice are currently being used by Mexicans as where approximately 40 million of people food, forage, ornamental and medicinal employ traditional remedies (García, 2002) plants. Tillandsia imperialis Morren ex which include a rich variety of medicinal Roezl (Bromeliaceae) is a tank epiphytic plants. Some of them are used for the treat- bromeliad found in tropical mountane cloud ment of respiratory infections, one of the forest, distributed throughout southeastern fi rst causes of hospitalization in Mexico Mexico, Salvador and Honduras. In Mexico and responsible for fi fty percent of deaths is known locally as “tencho”, and has been 22 Vite-Posadas, J.A. et al.: Morphoanatomical characterization and antimicrobial activity of Tillandsia imperialis widely exploited in fl orist trade. Infusions Pigmentation type was determined using the made from the leaves of T. imperialis are Benzing and Friedman’s protocol (1981). used in Mexican traditional medicine to treat respiratory ailments along with other Anatomical Characterization plants including Gardenia jasminoides, Physalis pubescens, Juglans regia, Hibis- Leaf paradermal and longitudinal fresh cus rosa-sinensis, and Rosa gallica as well sections of T. imperialis were prepared by as Dasypus novemcinctus (an armadillo), hand with single-edge razor blades. Other (Chino & Jácquez, 1986). samples were fi xed in FAA (Formalin, acetic acid, ethanol 100-50-500 mL in 1L), washed Scientifi c studies supporting the medicinal and dehydrated in increasing concentrations use of this plant and verifi ng its pharmaco- of ethanol solution (30°-50°-70°-96°-100°) logical effectiveness are lacking. The aim and included in Paraplast MR. Sections were of our work was to analyze comparative processed with the Johansen´s method morpho-anatomical characteristics of this (1940), and histochemical studies were bromeliad and evaluate its biological activ- carried out on some samples and were ob- ity, so corroborate the traditional uses of this served in a light microscope (Axiostar plus plant with experimental data. Carl Zeiss). MATERIAL AND METHODS Preparation of Extracts Plant Material Infl orescences and leaves were dried at am- bient temperature and powdered. Ten grams Tillandsia imperialis C.J. Morren ex Roezl of the powder were extracted three times 8h plants were bought within local markets of each one sequentially with organic solvents: Mexico City between February and April hexane, dichloromethane and methanol (J.K. 2004. The voucher specimen was depos- Baker) using a Soxhlet apparatus. The crude ited inside the herbarium of Universidad extracts were then evaporated to dryness Autónoma Metropolitana Iztapalapa in under vacuum conditions and the percentage Mexico (UAM-IZ), with the accession yield for each extract was determined. number 55702, (botanical identifi cation) by Ph.D. Adolfo Espejo (UAM-IZ). Microorganisms Evaluation Test Microorganisms Color Distribution and Foliar Pigmentation Six standard microorganisms strains were The color of bracts and leaves were com- used for testing antimicrobial activity: pared in vivo with the help of a standard Staphylococcus aureus (ATCC 29213), color handbook (Kornerup & Wanscher, Staphylococcus aureus OPS (clinical isolate; 1963). Lamina and sheath leaf pigmentation identifi ed and obtained from the Laboratorio was examined on foliar adaxial face as much de Microbiología, Centro de Investigaciones as foliar abaxial face. For detailed analysis, Biomédicas del Sur, IMSS), Staphylococcus each plant part (bract and leaf) was divided aureus AHD (clinical isolate), Streptococ- in three regions: apical, middle and basal. cus pyogenes (ATCC 08668), Escherichia 23 Núm. 31: 21-29 Marzo 2011 coli (ATCC 25922), Streptococcus faecalis RESULTS (ATCC 29212), Salmonella Typhi (ATCC 06539), Klebsiella pneumoniae (ATCC Color and Foliar Pigmentation 13883) y Candida albicans (ATCC