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Anatomy, Histochemistry, and Antifungal Activity of Anacardium humile (Anacardiaceae) Leaf

  • Vanessa de A. Royo (a1), Maria Olívia Mercadante-Simões (a2), Leonardo M. Ribeiro (a3), Dario A. de Oliveira (a4), Marcela Magda R. Aguiar (a1), Ellenhise R. Costa (a2) and Perácio Rafael B. Ferreira (a4)...


Leaves of Anacardium humile are used in Brazilian traditional medicine for the treatment of intestinal disturbances and skin lesions. This study aimed to define leaf diagnostic structural characters, to propose a new method of phytochemical analysis of secretions, prospect flavonoids and alkaloids, and to evaluate their inhibitory activity on Candida albicans. Common anatomical, phytochemical, and microbiological methods were used. Leaves of Anacardium occidentale were used as a reference for the structural analyses. The main structural characters were closed vascular system, absence of ducts at the medulla, bilateral mesophyll, absence of bundle sheath extension, and secretory idioblasts at the xylem. The ducts present schizogenous origin, and secreting activity is restricted to the initial phases of leaf blade expansion. The proposed new phytochemical method is practical and inexpensive and has potential for wide application. The abundance of tannins and flavonoids is related to medicinal use. A single peak in high-performance liquid chromatography indicated the presence of a pure substance not previously reported. The extract had a strong inhibitory effect on C. albicans. The obtained results confirm the potential of A. humile for the prospection of new bioactive compounds.


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Agra, M.F., Freitas, P.F. & Barbosa-Filho, J.M. (2007). Synopsis of the plants known as medicinal and poisonous in Northeast of Brazil. Braz J Pharmacognosy 17, 114140.
Ahmad, I. & Beg, A.Z. (2001). Antimicrobial and phytochemical studies on 45 Indian plants against multi-drug resistant human pathogens. J Ethnopharmacol 74, 113123.
Aligianis, N., Kalpoutzakis, E., Mitaku, S. & Chinou, I.B. (2001). Composition and antimicrobial activity of the essential oil of two Origanum species. J Agric Food Chem 49, 41684170.
Alvarenga, F.Q., Mota, B.C.F., Leite, M.N., Fonseca, J.M.S., Oliveira, D.A., Royo, V.A., Silva, M.L.A., Esperandim, V., Borges, A. & Laurentiz, R.S. (2013). In vivo analgesic activity, toxicity and phytochemical screening of the hydroalcoholic extract from the leaves of Psidium cattleianum Sabine. J Ethnopharmacol 150, 280284.
Álvarez, R., Encina, A. & Pérez Hidalgo, N. (2008). Pistacia terebinthus L. leaflets: An anatomical study. Plant Syst Evol 272, 107118.
Barbosa, W.L.R., Pinto, L.N., Quignard, E., Vieira, J.M.S., Silva, J.O.C. Jr. & Albuquerque, S. (2006). Arrabidaea chica (HBK) Verlot: Phytochemical approach, antifungal and trypanocidal activities. Braz J Pharmacognosy 18, 544548.
Barbosa-Filho, J.M., Piuvezam, M.R., Moura, M.D., Silva, M.S., Lima, K.V.B., Leitão-da-Cunha, E.V.L., Fechine, I.M. & Takemura, O.S. (2006). Anti-inflammatory activity of alkaloids: A twenty-century review. Braz J Pharmacognosy 16, 109139.
Brito, E.H.S., Fontenelle, R.O.S., Brilhante, R.S.N., Cordeiro, R.A., Monteiro, A.J., Sidrim, J.J.C. & Rocha, M.F.G. (2009). The anatomical distribution and antimicrobial susceptibility of yeast species isolated from healthy dogs. Vet J 182, 320326.
Cain, A.J. (1974). The use of Nile blue in the examination of lipids. Q J Microsc Sci 88, 383392.
Carrió, E. & Vallès, J. (2012). Ethnobotany of medicinal plants used in Eastern Mallorca (Balearic Islands, Mediterranean Sea). J Ethnopharmacol 141, 10211040.
Chaithra, M., Vivek, M.N., Asha, M.M., Kambar, Y., Prashith Kekuda, T.R. & Mallikarjun, N. (2013). Inhibitory effect of leaf and bark of Anacardium occidentale against clinical isolates of Staphylococcus aureus and Streptococcus mutans . J Drug Deliv Ther 3, 8083.
Chaves, M.H., Citó, A.M.G.L., Lopes, J.A.D., Costa, D.A., Oliveira, C.A.A., Costa, A.F. & Brito-Júnior, F.E.M. (2010). Total phenolics, antioxidant activity and chemical constituents from extracts of Anacardium occidentale L., Anacardiaceae. Braz J Pharmacognosy 20, 106112.
Christodoulakis, N.S., Mamoucha, S., Termentzi, A. & Fokialakis, N. (2015). Leaf structure and histochemistry of the hardy evergreen Euphorbia characias L. (Mediterranean spurge). Flora 210, 1318.
CLSI . (2012). Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically, Approved Standard, CLSI document M07-A9. Wayne, USA: Clinical and Laboratory Standards Institute.
Correia, S.J., David, J.P. & David, J.M. (2006). Metabólitos secundários de espécies de Anacardiaceae. Quim Nova 29, 12871300.
Das, S. & Rosazza, J.P.N. (2006). Microbial and enzymatic transformations of flavonoids. J Nat Prod 69, 499508.
David, R. & Carde, J.P. (1964). Coloration différentielle des inclusions lipidique et terpéniques des pseudophylles du Pin maritime au moyen du réactif nadi. CR Acad Sci D Nat 258, 13381340.
Ferreira, P.R.B., Mendes, C.S.O., Rodrigues, C.G., Rocha, J.C.M., Royo, V.A., Valério, H.M. & Oliveira, D.A. (2012). Antibacterial activity tannin-rich fraction from leaves of Anacardium humile . Cienc Rural 42, 18611864.
Feucht, W., Treutter, D., Dithmar, H. & Polster, J. (2008). Microspore development of three coniferous species: Affinity of nuclei for flavonoids. Tree Physiol 28, 17831791.
Franceschi, V.R. & Nakata, P.A. (2005). Calcium oxalate in plants: Formation and function. Annu Rev Plant Biol 56, 4171.
Freires, I.A. (2011). Atividade antifúngica de Schinus terebinthifolius (Aroeira) sobre cepas do gênero Candida . Rev Odontol Bras Central 20, 4145.
Furr, M. & Mahlberg, P.G. (1981). Histochemical analyses of lacticifers and glandular trichomes in Cannabis sativa . J Nat Prod 4, 153159.
Hasanuzzaman, M., Nahar, K., Alam, M.M., Roychowdhury, R. & Fujita, M. (2013). Physiological, biochemical, and molecular mechanisms of heat stress tolerance in plants. Int J Mol Sci 14, 96439684.
Havsteen, B.H. (2002). The biochemistry and medical significance of the flavonoids. Pharmacol Ther 96, 67202.
Johansen, D.A. (1940). Plant Microtechnique. New York, NY: McGraw-Hill Books.
Karnovsky, M.J. (1965). A formaldehyde-glutaraldehyde fixative of high osmolarity for use in electron microscopy. J Cell Biol 27, 137138.
Kingston, D.G.I. (2011). Modern natural products drug discovery and its relevance to biodiversity conservation. J Nat Prod 74, 496511.
Lacchia, A.P.S. & Carmello-Guerreiro, S.M. (2009). Aspectos ultra-estruturais dos canais secretores em órgãos vegetativos e reprodutivos de Anacardiaceae. Acta Bot Bras 23, 376388.
Lima, I.L.P., Scariot, A., Medeiros, M.B. & Sevilha, A.C. (2012). Diversidade e uso de plantas do Cerrado em comunidade de Geraizeiros no norte do estado de Minas Gerais, Brasil. Acta Bot Bras 23, 675684.
Mace, M.E. & Howell, C.R. (1974). Histochemistry and identification of condensed tannin precursor in roots of cotton seedlings. Can J Bot 52, 24232426.
Magro, A., Carolino, M., Bastos, M. & Mexia, A. (2006). Efficacy of plant extracts against stored-products fungi. Rev Iberoam Micol 23, 176178.
Martins, A.R., Soares, M.M., Redher, V.L.G., Bajay, M.M., Villela, P.M.S., Zucchi, M.I. & Appezzato-da-Glória, A. (2014). Use of anatomical, chemical, and molecular genetic characteristics in the quality control of medicinal species: A case study of sarsaparilla (Smilax spp.). Econ Bot 6, 410425.
Mauro, C., Pereira, A.M.S., Silva, C.P., Missima, J., Ohnuki, T. & Rinaldi, R.B. (2007). Anatomical study of species from savanna-like formation, Anemopaegma arvense (Vell.) Stellf. ex de Souza (catuaba), Zeyheria montana Mart. (bolsa-de-pastor) and Jacaranda decurrens Chamisso (caroba) – Bignoniaceae. Rev Bras Farmacognosy 17, 262265.
Mazia, D., Brewer, P.A. & Alfert, M. (1953). The cytochemical staining and measurement of protein with mercuric bromophenol blue. Biol Bull 104, 5767.
Mercadante-Simões, M.O., Mazzottini-dos-Santos, H.C., Nery, L.A., Ferreira, P.R.B., Ribeiro, L.M., Royo, V.A. & Oliveira, D.A. (2014). Structure, histochemistry and phytochemical profile of the bark of the sobol and aerial stem of Tontelea micrantha (Celastraceae – Hippocrateoideae). Ann Braz Acad Sci 86, 11671179.
Mercadante-Simões, M.O. & Paiva, E.A.S. (2013). Leaf colleters in Tontelea micrantha (Celastraceae, Salacioideae): Ecological, morphological and structural aspects. CR Biol 336, 400406.
Metcalfe, C.R. & Chalk, L. (1950). Anatomy of the Dicotyledons. Oxford: Clarendon Press.
Milani, J.F., Rocha, J.F. & Teixeira, S.P. (2012). Oleoresin glands in copaíba (Copaifera trapezifolia Hayne: Leguminosae), a Brazilian rainforest tree. Trees 26, 769775.
Montejano, H.A., Gervaldo, M. & Bertolotti, S.G. (2005). The excited-states quenching of resazurin and resorufin by p-benzoquinones in polar solvents. Dyes Pigments 64, 117124.
Mota, B.C.F., Royo, V.A.R., Fonseca, J.M.S., Santos, A.C., Melo, A.F. Jr., Menezes, E.V., Esperandim, V.R. & Laurentiz, R.S. (2015). Comparative studies between the chemical constituents and biological properties of the extracts from the leaves and barks of Myracrodruon urundeuva Fr. All. J Med Plant Res 9, 159168.
Namrita, L. & Navneet, K. (2014). Are plants used for skin care in South Africa fully explored? J Ethnopharmacol 153, 6184.
Nyanzi, R., Awouafack, M.D., Steenkamp, P., Jooste, P.J. & Eloff, J.N. (2014). Anticandidal activity of cell extracts from 13 probiotic Lactobacillus strains and characterisation of lactic acid and a novel fatty acid derivative from one strain. Food Chem 164, 470475.
O’Brien, T.P., Feder, N. & McCully, M.E. (1964). Polychromatic staining of plant cell walls by toluidine. Protoplasma 59, 368373.
Onasanwo, S.A., Fabiyi, T.D., Oluwole, F.S. & Olaleye, S.B. (2013). Analgesic and anti-inflammatory properties of the leaf extracts of Anacardium occidentalis in the laboratory rodents. Niger J Physiol Sci 27, 6571.
Palomino, J.C., Martin, A., Camacho, M., Guerra, H., Swings, J.A.N. & Portaels, S. (2002). Resazurin microtiter assay plate: Simple and inexpensive method. Antimicrob Agents Chemother 47, 36163619.
Paiva, E.A.S. & Machado, S.R. (2005). Role of intermediary cells in Peltodon radicans (Lamiaceae) in the transfer of calcium and formation of calcium oxalate crystals. Braz Arch Biol Technol 48, 147153.
Paiva, E.A.S., Pinho, S.Z. & Oliveira, D.M.T. (2011). Large plant samples: How to process for GMA embedding? In Light Microscopy: Methods and Protocols, Chiarini-Garcia, H. & Melo, R.C.N. (Eds.), pp. 3749). New York, NY: Springer/Humana Press.
Pearse, A.G.E. (1980). Histochemistry: Theoretical and Applied. Edinburgh: Churchill Livingstone.
Pizzolato, T.D. & Lillie, R.D. (1973). Mayers tannic acid-ferric chloride stain for mucins. J Histochem Cytochem 21, 5664.
Reis, A.L.L.E., Silva, D.S.S., Silva, K.L.F. & Chagas, D.B. (2014). Caracterização anatômica e histoquímica de raízes e plântulas de Anacardium occidentale L. (Anacardiaceae). Rev Arvore 38, 209219.
Rocha, M.S., Figueiredo, R.W., Araújo, M.A.M. & Moreira-Araújo, R.S.R. (2013). Physical and chemical characterization and antioxidant activity (in vitro) of fruit of the Piaui savanna. Rev Bras Frutic 35, 933941.
Sant’Anna-Santos, B.F., Thadeo, M., Meira, R.M.S.A. & Ascensão, L. (2006). Anatomia e histoquímica das estruturas secretoras do caule de Spondias dulcis Forst. (Anacardiaceae). Rev Arvore 30, 481489.
Shinde, V.M., Dhalwal, K., Potdar, M. & Mahadik, K.R. (2009). Application of quality control principles to herbal drugs. Int J Phytomed 1, 410.
Tona, L., Kambu, K., Ngimbi, N., Cimanga, K. & Vlietnick, A.J. (1998). Antiamoebic and phytochemical screening of some Congolese medical plants. J Ethnopharmacol 61, 5765.
Vidal, B.C. (1970). Dichroism in collagen bundles stained with xylidine-Ponceau 2R. Ann Histochim 15, 289296.
World Health Organization (1999). Monographs on Selected Medicinal Plants. Geneva: WHO.
Zayachkivska, O.S., Konturek, S.J., Drozdowicz, D., Konturek, P.C., Brzozowski, T. & Ghegotsky, M.R. (2005). Gastroprotective effects of flavonoids in plant extracts. J Physiol Pharmacol 56, 219231.
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Microscopy and Microanalysis
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