Anti-Candida activity of Tripodanthus acutifolius (Loranthaceae), mechanism of action and toxicity parameters


  • Gabriella da Rosa Monte Machado Universidade Federal do Rio Grande do Sul
  • Valéria Louzada Leal Universidade Federal do Rio Grande do Sul
  • Gabriella da Rosa Monte Machado Universidade Federal do Rio Grande do Sul
  • Mariéle Kliemann Universidade Federal de Ciências da Saúde de Porto Alegre
  • Mario Lettieri Teixeira Instituto Federal Catarinense
  • Rosana de Cássia de Souza Schneide Universidade de Santa Cruz do Sul
  • Chana de Medeiros da Silva Universidade de Santa Cruz do Sul
  • Alexandre Meneghello Fuentefria Universidade Federal do Rio Grande do Sul



antifungal agents, Candida spp., Tripodanthus acutifolius, synergism, fluconazole


The extensive use of azoles antifungals against infections caused by Candida species has been contributing to the selection of resistant strains to this antifungal class, demonstrating that the prospection of new antifungal agents is essential and urgent. Tripodanthus acutifolius (Loranthaceae) is a plant widely used in folk medicine with reported antimicrobial activity. In this context, this study aimed to evaluate the antifungal potential and the mechanisms of action of the crude methanolic extract (CME) of T. acutifolius leaves against Candida species, as well as evaluate their toxicity parameters. As results, the phytochemical characterization of CME suggested the presence of four phenolic compounds as well as a tripodantoside compound. The CME presented minimum inhibitory concentration (MIC) between 16 to 64 μg mL-1, with antifungal action possibly occurring in the fungal cell wall. In addition, the CME showed a synergic effect in combination with fluconazole (FLC). The CME demonstrated no mucosal irritation or tissue damage at all tested concentrations, as well as no cytotoxicity at the MIC values. This study is unprecedented and suggests that T. acutifolius is a new promising source for the development of anti-Candida agents on its own or as chemosensitizer associated to FLC.


ABOODY, M.S.A., MICKYMARAY, S. 2020. Anti-Fungal Efficacy and Mechanisms of Flavonoids. Antibiotics, 9(2): 45.

ANAND, J., RAI, N. 2017. Anticandidal synergistic activity of green tea catechins, antimycotics and copper sulphate as a mean of combinational drug therapy against candidiasis. J Mycol Med 27: 33-45.

ANTINORI. S., MILAZZO, L., SOLLIMA, S., GALLI, M., CORBELLINO, M. 2016. Candidemia and invasive candidiasis in adults: a narrative review. Eur J Intern Med 34: 21-28.

AREAL, N.A.S. 2015. Atualização do manejo da candidíase vulvovaginal (CVV) e da candidíase vulvovaginal recorrente (CVVR) visando à melhora da assistência a mulheres e gestantes. Monografia, Universidade Federal de Minas Gerais. Belo Horizonte, MG, Brazil.

ARENDRUP, M.C., PATTERSON, T.F. 2017. Multidrug-Resistant Candida: Epidemiology, Molecular Mechanisms and Treatment. J Infect Dis 216: S445–S451.

BADIEE, P., HASHEMIZADEH, Z. 2014. Opportunistic invasive fungal infections: diagnosis & clinical management. Indian J Med Res 139: 195-204.

BRASIL. 2000. Instrução Normativa n° 3/2000: aprova Regulamento Técnico de Métodos de Insensibilização para Abate Humanitário de Animais de Açougue. Ministério da Agricultura.

CASTRO, M.D.L., AYUSO, L.E.G. 2000. Soxhlet Extraction. In: Wilson ID Encyclopedia of Separation Science, Academic Press: 2701-2709.

CHAUDHARI, S.P., BANGAR, J.V., AKUSKAR, G.K., RATNAPARKHI, M.P. 2014. Development and validation of UV spectrophotometric method for simultaneous estimation of rutin and quercetin in niosome formulation. Der Pharmacia Lettre 6: 271-276.

CLINICAL AND LABORATORY STANDARDS INSTITUTE, CLSI. 2008. Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeasts, Approved Standard -Third Edition, CLSI document M27-A3. Wayne, PA: Clinical and Laboratory Standards Institute.

CLINICAL AND LABORATORY STANDARDS INSTITUTE, CLSI. 2012. Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeasts, Fourth Informational Supplement, CLSI document M27-S4. Wayne, PA: Clinical and Laboratory Standards Institute.

COELHO, R.P., FEKSA, D.L., OLIVEIRA, P.M., GÜLLICH, A.A.C., PILAR, B.C., POCCOLI, J.C.E., MANFREDINI, V. 2018. Protective effect of the hydroalcoholic extract of Tripodanthus acutifolius in hypercholesterolemic Wistar rats. Biomed Pharmacother 97: 300-309.

DAUD, A., GALLO, A., RIERA, A.S. 2005. Antimicrobial properties of Phrygilanthus acutifolius. J Ethnopharmacol 99: 193-197.

DAUD, A., HABIB, N., RIERA, A.S. 2006. Anti-inflammatory, anti-nociceptive and antipyretic effects of extracts of Phrygilanthus acutifolius flowers. J Ethnopharmacol 108: 198-203.

DE OLIVEIRA FILHO, A.A., DE OLIVEIRA, H.M.B.F., DE SOUSA, J.P., MEIRELES, D.R.P., DE AZEVEDO MAIA, G.L., FILHO, J.M.B., DE SIQUEIRA JÚNIOR, J.P., LIMA, E.O. 2016. In vitro anti-Candida activity and mechanism of action of the flavonoid isolated from Praxelis clematidea against Candida albicans species. Journal of Applied Pharmaceutical Science 6(01): 066-069. 10.7324/JAPS.2016.600111

DEORUKHKAR, S.C., SAINI, S., MATHEW, S. 2014. Non-albicans Candida Infection: An Emerging Threat. Interdiscip Perspect Infect Dis.

ESCALANTE, A., GATTUSO, M., PEREZ, P., ZACCHINO, S. 2008. Evidence for the mechanism of action of the antifungal Phytolaccoside B Strainstraind from Phytolacca tetramera Hauman. J. Nat. Prod 71: 1720–1725.

ESCHENAUER, G.A., NGUYEN, M.H., CLANCY, C.J. 2015. Is fluconazole or an echinocandin the agent of choice for candidemia. Ann Pharmacother 49: 1068-1074.

FRESHNEY, R.I. 2005. Culture of Animal Cells: a Manual of Basic Technique, 5th Ed. Hoboken NJ, John Wiley & Sons.

GAITÉN, Y.I.G., MARTÍNEZ, M.M., HENRIQUES, A.T., ALONSO, G.D.B. 2010. Análisis de flavonoides en una fracción butanólica obtenida de Phyllanthus orbicularis HBK. Rev Cubana Farm 44: 367-373.

GRÜNER, J.M., SOUZA, T.K., BENITEZ, L.B., SILVA, C.M. 2012. Análise do perfil fitoquímico de Tripodanthus acutifolius (Ruiz & Pavón) Tieghem, Loranthaceae. Revista Jovens Pesquisadores 1: 9-17.

INTERAGENCY COORDINATING COMMITTEE ON THE VALIDATION OF ALTERNATIVE METHODS, ICCVMA. 2010. The Hen's Egg Test–Chorioallantoic Membrane (HET-CAM) Test Method. Research Triangle Park: National Toxicology Program.

INTERNATIONAL ORGANIZATION FOR STANDARDIZATION, ISO 10993-5. 2009. Biological Evaluation of Medical Devices, Part 5: Tests for In Vitro Cytotoxicity. International Organization for Standardization: Geneva, Switzerland.

INTERSIMONE, N.H., THOENE, A.D., RIERA, A.S. 2005. Efecto diurético de extractos acuosos y alcohólicos de flores de Phrygilanthus acutifolius (corpo) en ratas. Revista Cubana Plantas Medicinales 10: 3-4.

JOHNSON, M.D., MACDOUGALL, C., OSTROSKY-ZEICHNER, L., PERFECT, J.R., REX, J.H. 2004. Combination antifungal therapy. Antimicrob Agents Chemother 48: 693-715.

KUMAR, A., ZARYCHANSKI, R., PISIPATI, A., KUMAR, A., KETHIREDDY, S., BOW, E.J. 2018. Fungicidal versus fungistatic therapy of invasive Candida infection in non-neutropenic adults: a meta-analysis. Mycology 9: 116-128.

LEE, H.S., KIM, Y. 2017. Paeonia lactiflora inhibits cell wall synthesis and triggers membrane depolarization in Candida albicans. J. Microbiol. Biotechnol 27(2): 395-404.

LIU, W., LI, L.P., ZHANG, J.D., LI, Q., SHEN, H., CHEN, S.M., HE, L.J., YAN, L., XU, G.T., AN, M.M., JIANG, Y.Y. 2014. Synergistic Antifungal Effect of Glabridin and Fluconazole. PLoS ONE 9(7): e103442.

LU. M., LI, T., WAN, J., LI, X., YUAN, L., SUN, S. 2016. Antifungal effects of phytocompounds on Candida species alone and in combination with fluconazole. International journal of antimicrobial agents 49(2): 125-136.

MACHADO, G.R.M., PIPPI, B., LANA, D.F.D., AMARAL, A.P.S., TEIXEIRA, M.L., SOUZA, K.C., FUENTEFRIA, A.M. 2016. Reversal of fluconazole resistance induced by a synergistic effect with Acca sellowiana in Candida glabrata strains. Pharm Biol 54: 2410–2419

MARTINS, N., BARROS, L., HENRIQUES, M., SILVA, S., FERREIRA, I.C.F.R. 2015. Activity of phenolic compounds from plant origin against Candida species. Ind Crop Prod 74: 648-670.

MEIRELLES, G.C., PIPPI, B., HATWIG, C., BARROS, F.M.C., OLIVEIRA, L.F.S., VON POSER, G.L., FUENTEFRIA, A.M. 2017. Synergistic antifungal activity of the lipophilic fraction of Hypericum carinatum and fluconazole. Rev Bras Farmacogn 7: 118-123.

MESA-ARANGO, A.C., MONTIEL-RAMOS, J., ZAPATA, B., DURÁN, C., BETANCUR-GALVIS, L., STASHENKO, E. 2009. Citral and carvone chemotypes from the essential oils of Colombian Lippia alba (Mill.) N.E. Brown: composition, cytotoxicity and antifungal activity. Memórias do Instituto Oswaldo Cruz 104: 6.

MOSMANN, T. 1983. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65: 55-63.

NEILSON, A.P., GREEN, R.J., WOOD, K.V., FERRUZZI, M.G. 2006. High-throughput analysis of catechins and theaflavins by high performance liquid chromatography with diode array detection. J Chromatogr A 1132: 132-140.

O’NEILL, J. 2014. Antimicrobial resistance: tackling a crisis for the health and wealth of nations. Review on Antimicrobial Resistance. Accessed 15 Jan 2018

PAWAR, N.P., SALUNKHE, V.R. 2013. Development and validation of UV spectrophotometric method for simultaneous estimation of rutin and gallic acid in hydroalcoholic extract of Triphala churna. Int J Pharmtech Res 5: 724-729.

PFALLER, M.A., ANDES, D.R., DIEKEMA, D.J., HORN, D.L., REBOLI, A.C., ROTSTEIN, C., FRANKS, B., AZIE, N.E. 2014. Epidemiology and Outcomes of Invasive Candidiasis Due to Non-albicans Species of Candida in 2,496 Patients: Data from the Prospective Antifungal Therapy (PATH) Registry 2004–2008. PLoS ONE 9: e101510.

PIPPI, B., LANA, A.J.D., MORAES, R.C., GÜEZ, C.M., MACHADO, M., DE OLIVEIRA, L.F., LINO VON POSER, G., FUENTEFRIA, A.M. 2015. In vitro evaluation of the acquisition of resistance, antifungal activity and synergism of brazilian red propolis with antifungal drugs on Candida spp. J Appl Microbiol 118: 839-850.

PRASAIN, J.K., BARNES, S. 2014. Uptake and metabolism of dietary proanthocyanidins. In: Watson RR, Preedy VR, Zibadi S Polyphenols in human health and disease. Polyphenols in Human Health and Disease 1: 553-560.

PRETSCH, E., CLERC, T., SEIBL, J., SIMON, W. 1989. Tables of spectral data for structure determination of organic compounds. Laboratory Chemical Practice, 2nd Ed. Springer-Verlag Berlin Heidelberg.

RICCO, R.A., LOZANO, V., LATOR, V., GURNI, A.A., WAGNER, M.L. 2008. Modificación en el perfil de polifenoles de Tripodanthus acutifolius (Ruiz et Pav.) Tiegh. (Loranthaceae) infectado por “Cochinilla” (Homoptera - Lecanidae). Lat Am J Pharm 27: 258-262.

SELEEM, D., PARDI, V., MURATA, R.M. 2017. Review of flavonoids: A diverse group of natural compounds with anti-Candida albicans activity in vitro. Archives of Oral Biology 76: 76–83. .

SILVA, C.M. 2014. Estudo químico e biológico de Tripodanthus acutifolius (Ruiz & Pav.) Tiegh., Loranthaceae. Dissertation, Universidade Federal do Rio Grande do Sul. Porto Alegre, RS. Brazil.

SILVA, C.R., NETO, J.B.A., CAMPOS, R.S., FIGUEIREDO, N.S., SAMPAIO, L.S., MAGALHÃES, H.I., CAVALCANTI, B.C., GASPAR, D.M., DE ANDRADE, G.M., LIMA, I.S., DE BARROS VIANA, G.S., DE MORAES, M.O., LOBO, M.D., GRANGEIRO, T.B., NOBRE JÚNIOR, H.V. 2014. Synergistic effect of the flavonoid catechin, quercetin, or epigallocatechin gallate with fluconazole induces apoptosis in Candida tropicalis resistant to fluconazole. Antimicrob Agents Chemother 58: 1468-1478. 10.1128/AAC.00651-13

SILVA, D., DINIZ-NETO, H., CORDEIRO, L., SILVA-NETA, M., SILVA, S., ANDRADE-JÚNIOR, F., LEITE, M., NÓBREGA, J., MORAIS, M., SOUZA, J., ROSA, L., MELO, T., SOUZA, H., SOUSA, A., RODRIGUES, G., OLIVEIRA-FILHO, A., LIMA. E. 2020. (R)-(+)-β-Citronellol and (S)-(−)-β-Citronellol in combination with Amphotericin B against Candida Spp. International Journal of Molecular Sciences 21(5): 1785.

SILVERSTEIN, R.M., WEBSTER, F.X., KIEMLE. D.J. 2007. Identificação espectrométrica de compostos orgânicos. Rio de Janeiro: Sétima edição. LTC – Livros Técnicos e Científicos Editora S.A.

SIMONETTI, E., ETHUR, M.E., CASTRO, L.C., KAUFFMANN, C., GIACOMIN, A.C., LEDUR, A., AROSSI, K., PACHECO, L.A., GOETTERT, M.I., FALEIRO, D., FREITAS, E.M. 2016. Avaliação da atividade antimicrobiana de extratos de Eugenia anomala e Psidium salutare (Myrtaceae) frente à Escherichia coli e Listeria monocytogenes. Rev Bras Plantas Med 18: 9-18.

SOBERÓN, J.R., SGARIGLIA, M.A., SAMPIETRO, D.A., QUIROGA, E.N., VATTUONE, M.A. 2007. Antibacterial activity of plant extracts from northwestern Argentina. J Appl Microbiol 102: 1450-1461.

SOBERÓN, J.R., SGARIGLIA, M.A., SAMPIETRO, D.A., QUIROGA, E.N., VATTUONE, M.A. 2010a. Study of antiinflammatory activity of metabolites strainstraind from Tripodanthus acutifolius. Molecular Medicinal Chemistry 21: 88-90.

SOBERÓN, J.R., SGARIGLIA, M.A., SAMPIETRO, D.A., QUIROGA, E.N., VATTUONE, M.A. 2010b. Free radical scavenging activities and inhibition of inflammatory enzymes of phenolics strainstraind from Tripodanthus acutifolius. J Ethnopharmacol 130: 329-333.

SOBERÓN, J.R., SGARIGLIA, M.A., SAMPIETRO, D.A., QUIROGA, E.M., SIERRA, M.G., VATTUONE, M.A. 2010c. Purification and identification of antibacterial phenolics from Tripodanthus acutifolius leaves. J Appl Microbiol 108: 1757-1768.

SOBERÓN, J.R., SGARIGLIA, M.A., SAMPIETRO, D.A., VATTUONE, M.A. 2011. Actividad antifúngica combinada de tripodantósido y anfotericina B sobre levaduras del genero Candida. Congreso. XXVIII Jornadas Científicas de la Asociación de Biología de Tucumán.

SOUZA, T.K., SILVA, C.M., BENITEZ, L.B., VON POSER, G.L., ZUANAZZI, J.A.S. 2014. Atividade antibacteriana do extrato aquoso de Tripodanthus acutifolius frente à Staphylococcus aureus. Revista Jovens Pesquisadores 4: 6-18.

TREMÉA, C.R. 2015. Atividade dos óleos essenciais de Litsea cubeba e Cymbopogon martini sobre isolados do complexo Cryptococcus neoformans. Thesis, Universidade Federal de Goiás. Goiânia, GO, Brazil.

UPPULURI, P., KHAN, A., EDWARDS, J.E. 2017. Current Trends in Candidiasis. In: Prasad R. Candida albicans: Cellular and molecular biology, 2nd Ed. Springer International Publishing 2: 5-23.

VALLABHANENI, S., CLEVELAND, A.A., FARLEY, M.M., HARRISON, L.H., SCHAFFNER, W., BELDAVS, Z.G., DERADO, G., PHAM, C.D., LOCKHART, S.R., SMITH, R.M. 2015. Epidemiology and risk factors for echinocandin non susceptible Candida glabrata bloodstream infections: data from a large multisite population-based candidemia surveillance program, 2008–2014. Open Forum Infect Dis 2: ofv163.

WAGNER, H., ULRICH-MERZENICH, G. 2009. Synergy research: approaching a new generation of phytopharmaceuticals. Phytomedicine 16: 97-110.

WAGNER, M.L., RICCO, R.A., RANEA, F.G., GURNI, A.A. 2009. Flavonoides de especies argentinas del género Tripodanthus (Eichl.) Tiegh. (Loranthaceae). Dominguezia 25: 21-27.

WANG, Y.H., DONG, H.H., ZHAO, F., WANG, J., YAN, F., JIANG, Y.Y., JIN, Y.S. 2016. The synthesis and synergistic antifungal effects of chalcones against drug resistant Candida albicans. Bioorganic & medicinal chemistry letters 26(13): 3098-3102.

WHALEY, S.G., BERKOW, E.L., RYBAK, J.M., NISHIMOTO, A.T., BARKER, K.S., ROGERS, P.D. 2017. Azole antifungal resistance in Candida albicans and emerging non-albicans Candida species. Front Microbiol 7: 2173.

ZIDA, A., BAMBA, S., YACOUBA, A., OUEDRAOGO-TRAORE, R., GUIGUEMDÉ, R.T. 2016. Anti-Candida albicans natural products, sources of new antifungal drugs: A review. J Mycol Med 27: 1-19.



Como Citar

Machado, G. da R. M., Leal, V. L., Machado, G. da R. M. ., Kliemann, M., Teixeira, M. L. ., Schneide, R. de C. de S. ., Silva, C. de M. da ., & Fuentefria, A. M. (2021). Anti-Candida activity of Tripodanthus acutifolius (Loranthaceae), mechanism of action and toxicity parameters. Scientific Electronic Archives, 14(6), 37–48.



Biological Science