Red de Revistas Científicas de América Latina, el Caribe, España y Portugal Sistema de Información Científica SILVA TRENTIN, DANIELLE; RIGON ZIMMER, KARINE; VANUSA SILVA, MÁRCIA; BRANDT GIORDANI, RAQUEL; MACEDO, ALEXANDRE JOSÉ MEDICINAL PLANTS FROM BRAZILIAN CAATINGA: ANTIBIOFILM AND ANTIBACTERIAL ACTIVITIES AGAINST Pseudomonas aeruginosa Revista Caatinga, vol. 27, núm. 3, julio-septiembre, 2014, pp. 264-271 Universidade Federal Rural do Semi-Árido Mossoró, Brasil Available in: http://www.redalyc.org/articulo.oa?id=237132104030 Revista Caatinga, ISSN (Printed Version): 0100-316X [email protected] Universidade Federal Rural do Semi-Árido Brasil How to cite Complete issue More information about this article Journal's homepage www.redalyc.org Non-Profit Academic Project, developed under the Open Acces Initiative Universidade Federal Rural do Semi-Árido ISSN 0100-316X (impresso) Pró-Reitoria de Pesquisa e Pós-Graduação ISSN 1983-2125 (online) http://periodicos.ufersa.edu.br/index.php/sistema MEDICINAL PLANTS FROM BRAZILIAN CAATINGA: ANTIBIOFILM AND AN- TIBACTERIAL ACTIVITIES AGAINST Pseudomonas aeruginosa 1 DANIELLE SILVA TRENTIN 2,3 , KARINE RIGON ZIMMER 2,3 , MÁRCIA VANUSA SILVA 4, RAQUEL BRANDT GIORDANI 5, ALEXANDRE JOSÉ MACEDO 2,3* ABSTRACT - The Caatinga biome covers a vast area in northeastern Brazil and presents a high level of biodi- versity. It is known that about 400 plant species are used by semi-arid local communities for medical purposes. Based on ethnopharmacological reports, this study aims to screen 24 species from Caatinga regarding the abil- ity to prevent biofilm formation and to inhibit the growth of Pseudomonas ae ruginosa - a major opportunistic human pathogen and an important causative agent of morbidity and mortality. The effects of aqueous extracts, at 0.4 and 4.0 mg mL -1, on biofilm formation and on growth of P. aeruginosa ATCC 27853 were studied using the crystal violet assay and the OD 600 absorbance, respectively. The most active extracts were analyzed by thin- layer chromatography and high performance liquid chromatography. Our investigation pointed extracts of four species with potential application for the control of P. aeruginosa : Anadenanthera colubrina (Vell.) Brenan, Commiphora leptophloeos (Mart.) J.B. Gillett, Myracrodruoun urundeuva Allemão, whose antibiofilm effects (89%, 56% and 79% inhibition of biofilm, respectively) were associated with complete inhibition of bacterial growth, and Pityrocarpa moniliformis (Benth.) Luckow & R.W. Jobson, which were able avoid 68% of biofilm formation and inhibited 30% bacterial growth. The qualitative phytochemical analyses reveal the complexity of the samples as well as the presence of compounds with high molecular weight. Keywords : Biofilm. Anadenanthera colubrina . Commiphora leptophloeos. Myracrodruoun urundeuva. Pityro- carpa moniliformis . Traditional medicine. PLANTAS MEDICINAIS DA CAATINGA BRASILEIRA: ATIVIDADES ANTIBIOFILME E ANTI- BACTERIANA CONTRA Pseudomonas aeruginosa 1 RESUMO - O bioma Caatinga abrange uma vasta área no nordeste do Brasil e apresenta uma expressiva biodi- versidade. Sabe-se que aproximadamente 400 espécies de plantas são utilizadas por comunidades locais para fins medicinais. Com base em relatos etnofarmacológicos, este estudo tem por objetivo rastrear 24 espécies de plantas da Caatinga quanto à capacidade de impedir a formação de biofilme e de inibir o crescimento de Pseu- domonas aeruginosa - importante patógeno oportunista humano e agente causador de morbidade e mortalidade. Os efeitos dos extratos aquosos, nas concentrações de 0,4 e 4,0 mg mL -1, sobre a formação de biofilme e o cres- cimento de P. aeruginosa ATCC 27853 foram avaliados através do ensaio de cristal violeta e da densidade óptica a 600 nm, respectivamente. Os extratos mais ativos foram analisados por cromatografia em camada del- gada e cromatografia líquida de alta eficiência. Nossa investigação indicou extratos de quatro espécies com potencial aplicação para o controle de P. aeruginosa : Anadenanthera colubrina (Vell.) Brenan, Commiphora leptophloeos (Mart.) J.B. Gillett, Myracrodruoun urundeuva Allemão, cujo efeitos antibiofilme (89%, 56% e 79% de inibição de biofilme, respectivamente) foram associados com a completa inibição do crescimento bac- teriano, e, Pityrocarpa moniliformis (Benth.) Luckow & R.W. Jobson, a qual foi capaz de evitar 68% da forma- ção de biofilme e inibiu em 30% o crescimento bacteriano. As análises qualitativas fitoquímicas indicam hete- rogeneidade entre as amostras, bem como a presença de compostos com elevado peso molecular. Palavras-chave : Biofilme. Anadenanthera colubrina . Commiphora leptophloeos. Myracrodruoun urundeuva. Pityrocarpa moniliformis. Medicina tradicional. ___________________ *Corresponding author 1 Received for publication at 10/06/2013; accepted at 06/05/2014. 2Faculdade de Farmácia, UFRGS, Av. Ipiranga, 2752, 90610-000, Porto Alegre-RS, Brazil; [email protected], karorzim- [email protected], [email protected]. 3Centro de Biotecnologia, , UFRGS, Av. Bento Gonçalves, 9500, 91509-900, Porto Alegre-RS, Brazil. 4Centro de Ciências Biológicas e Departamento de Bioquímica, UFPE, Av. Prof. Moraes Rego, 1235 - Cidade Universitária, 50670-901, Recife-PE, Brazil; [email protected]. 5Centro de Ciências da Saúde, Departamento de Farmácia, UFRN, R. General Gustavo Cordeiro de Farias, sn, 59010-180, Natal-RN, Bra- zil; [email protected]. Revista Caatinga , Mossoró, v. 27, n. 3, p. 264 – 271, jul. – set., 2014 264 MEDICINAL PLANTS FROM BRAZILIAN CAATINGA: ANTIBIOFILM AND ANTIBACTERIAL ACTIVITIES AGAINST Pseudomonas aeruginosa D. S. TRENTIN et al. INTRODUCTION (RICHARDS et al. 1999) – a usually biofilm- associated infection. The folk medicine has a great contribution to Targeting bacterial virulence (like biofilm human health care long before modern medicine formation) is an alternative approach to antibacterial began. Nowadays, the use of medicinal plants as a therapy that offers promising opportunities to inhibit source of remedies remains common to the medical pathogenesis without threatening bacterial existence, traditions of many cultures and there is a strong evi- resulting in a reduced selection pressure for drug- dence of integration between traditional and modern resistant mutations (CEGELSKI et al., 2008). Plants methods of health care. Understanding the practices have been used as medicine in all cultures and the of the past which are maintained by the traditional interest in research involving antimicrobial activity knowledge passed generation by generation, it pro- and medicinal properties of plants has increased. vides insight into and may lead to improvements in Given the great importance of biofilm formation in actual pharmaceutical practice (VANDEBROEK et the infectious process, the association of screening of al., 2011, MEDEIROS; ALBUQUERQUE, 2012). medicinal plants and the modulation of P. aerugi- Therefore, there is an urgent call to collect informa- nosa biofilm started to be considered (DING et al., tion on and to identify the best health practices all 2011). We used aqueous extracts to reproduce the over the world, and use them cost-effectively to en- traditional medicine preparation, and thereby, it en- hance health care delivery (ORTEGA, 2006). ables us to evaluate their effectiveness, considering The Caatinga, typical semi-arid vegetation, in vitro antibacterial/antibiofilm properties against represents the fourth largest area covered by a single this important pathogen. In this context, we investi- vegetation form in Brazil and accounts account for gated 24 species of plants from Caatinga vegetation about 60% of the northeast territory (ANDRADE- (45 aqueous extracts) for their ability to inhibit LIMA, 1981, SAMPAIO et al., 2002). Several publi- planktonic growth and to prevent biofilm formation cations describe the rich flora in this region as hav- by P. aeruginosa . Also, we performed preliminary ing many medicinal purposes and great phytochemi- phytochemical analysis of the most active extracts. cal potential (AGRA et al., 2007; ALBUQUERQUE et al., 2007a; 2012). In addition to being widely known and used by local communities, many medici- MATERIALS AND METHODS nal species in the Caatinga are sold as herbal prod- ucts (ALBUQUERQUE et al., 2007b; CARTAXO et The plants were collected at the Parque Na- al., 2010; MONTEIRO et al., 2011). cional do Catimbau (PARNA from Catimbau), Per- Pseudomonas aeruginosa is a major oppor- nambuco State, Brazil (8°30'0.07"S and 37° tunistic human pathogen and an important causative 26'41.00"W), in July and August 2009. The species agent of morbidity and mortality. In addition antim- were chosen according to the published ethnophar- icrobial resistance of P. aeruginosa remains a chal- macological data and using information from the lenge for clinical treatment, antibiotic selection and local community. The taxonomic identifications effectiveness. This bacterium is able to express all were performed by the MSc. Alexandre Gomes da known mechanisms of antimicrobial resistance, such Silva from the Federal University of Pernambuco, as the low outer membrane permeability, the expres- Pernambuco, Brazil. Voucher specimens were de- sion of broad multidrug efflux systems and the hori- posited in the herbarium Dárdano de Andrade-Lima zontal gene transfer (STRATEVA; YORDANOV, at Agronomic Institute of Pernambuco (IPA), Brazil 2009). Therefore, P. aeruginosa often exhibits a (Table 1). Aqueous extracts were prepared by static multidrug-resistant