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Evaluation of the Toxicity and Molluscicidal and Larvicidal Activities of Schinopsis Brasiliensis Stem Bark Extract and Its Fractions

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Evaluation of the Toxicity and Molluscicidal and Larvicidal Activities of Schinopsis Brasiliensis Stem Bark Extract and Its Fractions Rev Bras Farma; 24(2014): 298-303 Original article Evaluation of the toxicity and molluscicidal and larvicidal activities of Schinopsis brasiliensis stem bark extract and its fractions Clisiane C.S. Santosa, Silvan S. Araújoa, André L.L.M. Santosa, Elis C.V. Almeidaa, Antônio S. Diasa, Nicole P. Damascenaa, Deisylaine M. Santosa, Matheus I.S. Santosa, Karlos A.L.R. Júniorb, Carla K.B. Pereirab, Amanda C.B. Limaa, Andrea Y.K.V. Shana, Antônio E.G. Sant’anab, Charles S. Estevama, Brancilene S. Araujoa,* aLaboratório de Bioquímica e Química de Produtos Naturais, Departamento de Fisiologia, Universidade Federal de Sergipe, São Cristóvão, SE, Brazil bLaboratório de Química de Produtos Naturais, Departamento de Química, Universidade Federal de Alagoas, Maceió, AL, Brazil. ARTICLE INFO ABSTRACT Article history: Dengue fever and schistosomiasis are major public health issues for which vector control Received 7 October 2013 using larvicide and molluscicide substances present in plants provides a promising strategy. Accepted 5 March 2014 This study evaluated the potential toxicity of the extract of hydroethanol Schinopsis brasiliensis Engl., Anacardiaceae, stem bark and its chloroform, hexane, ethyl acetate, and hydromethanol Keywords: fractions against Artemia salina and Aedes Aegypti larvae and snails Biomphalaria glabrata. All Artemia salina of the assays were performed in triplicate and the mean mortality rates were used to determine Aedes the LC50 and LC90 values using the probit method. The hydroethanol hydromethanol extract Biomphalaria and fraction were free of toxicity towards A. salina (LC50 > 1000 μg/ml), while chloroform Larvicide fraction was moderately toxic (LC50 313 μg/ml); ethyl acetate and hexane fractions displayed Molluscicide low toxicity, with LC50 557 and 582 μg/ml, respectively. Chloroform, hexane, and ethyl acetate Schinopsis brasiliensis fractions showed larvicidal potential towards A. aegypti (LC50 values of 345, 527 and 583 μg/ml, respectively), while chloroform and ethyl acetate fractions were highly toxic to B. glabrata (LC90 values of 68 and 73 μg/ml, respectively). Based on these findings, ethyl acetate, chloroform, and hexane fractions should be further investigated for their potential use against the vectors of dengue and schistosomiasis. © 2014 Sociedade Brasileira de Farmacognosia. Published by Elsevier Editora Ltda. All rights reserved. Introduction concern because it is largely dispersed in urban areas (Porto et al., 2008). Given that 50 to 80 million people are infected Aedes aegypti L. is the vector responsible for transmitting with dengue anually in over 100 countries, vector control is the tropical viral infection known as dengue, dengue fever, extremely important and consists of eliminating breeding or breakbone fever. It is considered a major public health sites and applying insecticides (Lingon, 2005; Mendonça et * Corresponding author. E-mail: [email protected] (B.S. Araújo). 0102-695X/$ - see front matter © 2014 Sociedade Brasileira de Farmacognosia. Published by Elsevier Editora Ltda. All rights reserved. http://dx.doi.org/ 10.1016/j.bjp.2014.07.006 Clisiane C.S. Santos et al. / Rev Bras Farmacogn 24(2014): 298-303 299 al., 2009). However, synthetic insecticides have been observed Cristina Farias and a voucher specimen was deposited in to have low efficiency due to resistance of insect populations the herbarium of the Department of Biology of the Federal (Barreto, 2005). University of Sergipe under the registration voucher number Schistosomiasis is another major problem caused by 24442 ASE. transmission vectors. According to the World Health Organization (WHO), this ancient disease affects indiviuals Preparation of the hydroethanol extract and its fractions over 240 million in over 78 countries (WHO, 2013). Schistosoma mansoni is the flatworm parasite responsible for most cases The steam bark of S. brasiliensis was dried at room of infection. In the countries of South America and Central temperature, reduced to powder using a slicer, and then America, primarily S. mansoni uses Biomphalaria glabrata extracted by cold maceration with 90% ethanol for five snails of the host to an intermediate to complete its life cycle days. The equipament was then filtered and concentrated (Rey, 2010). Among the methods used to reduce occurrences on a rotatory evaporator (LS Logen Scientific, Lagos, Nigeria) of the disease, one promising strategy is the control of under reduced pressure at 45°C to produce the hydroethanol populations because malacological eliminating the vector extract (HEE). The extract (40 g) was dissolved in methanol: with substances endowed with molluscicide properties water (40% v/v) and successively subjected to liquid-liquid interrupts the life cycle of the parasite, and thereby, disease extraction with hexane, chloroform, ethyl acetate and hexane transmission (Santos et al., 2010). to obtain infor (HxF), chloroform (ChlF), ethyl acetate (EAF), Dengue fever and schistosomiasis are two major public and hydromethanol (HMF) fractions. health problems that many countries, including Brazil, are seeking to solve. In this context, the use of plants to control Phytochemical screening or eradicate these and other diseases is premised on their availability and lower environmental impact, which are Extracts and fractions (5 ml, 1 mg/ml) were qualitatively variables for obtaining an effective natural product. Plants analyzed by precipitation and colorimetric methods described are important sources of biologically active products that by Matos (2009) to detect anthocyanins, anthocyanidins, can be responsible for various biological actions, including aurones, chalcones, flavanols, flavones, flavonols and the larvicidal and molluscicidal properties shown in several xanthones [pH-related color variation by adding sodium studies (Prophiro et al., 2008; Santos et al., 2012). hydroxide (3 mol/l) and hydrochloric acid (1 mol/l)], A previous survey of different parts of plants from leucoanthocyanidins and catechins (acid-base reactions northeastern Brazil revealed a high toxicity towards A. aegypti followed by heating), tannins [ferric chloride precipitation and B. glabrata (Luna et al., 2005; Oliveira et al., 2006). Although (1 mol/l)], steroids and triterpenoids (Lieberman-Buchard the chemical constituents responsible for causing this toxicity reaction), saponins (Lieberman-Buchard reaction and foam have not been identified, the pharmacological effects of these formation) and alkaloids (Dragendorff reaction). plants, e.g., molluscicidal and larvicidal activities, are known to be related to the presence of secondary metabolites such Toxicity assay as tannins, saponins, terpenoids, steroids, and flavonoids (Treyvaud et al., 2000; Alcanfor et al., 2001; Hymete et al., 2005; The toxic activity of the extract and fractions of S. brasiliensis Cantanhede et al., 2010). was assessed by the lethality test against Artemia salina Leach Schinopsis brasiliensis Engl., Anacardiaceae, popularly known as described by Meyer et al. (1982) with some modifications. as “baraúna,” “braúna,” or “quebracho,” is used in folk medicine A. salina eggs were incubated in cotton-filtered seawater at to treat various diseases (Almeida et al., 2010). Biological studies room temperature for 24 h, after which the larvae were collected using a methanol leaf extract from S. brasiliensis demonstrated with the aid of a light source to attract them. Approximately its antioxidant, antimicrobial, and anti-inflammatory activities ten nauplii were transferred to ELISA plate wells containing (Ferreira-Junior et al., 2011; Saraiva et al., 2011), while the 1 ml of HEE and the fraction concetrations of solutions at 1, 10, extract from the plant seed hydroethanol has been shown to 100, and 1000 μg/ml prepared in seawater containing 1% DMSO. have toxic effects against A. aegypti larvae (Souza et al., 2011). The control group consisted of only the solvent and larvae, and Therefore, the goal of this study was to evaluate a stem bark all assays were performed in triplicate. The dead larvae were extract from S. brasiliensis and its fractions for toxicity and counted after exposure for 24 h. molluscicidal and larvicidal activities. Larvicidal assay Materials and methods Third stage larvae were A. aegypti mosquitoes used to evaluate the larvicidal activity of the HEE and its fractions Collection and identification of plant material using the methodology described by the World Health Organization (WHO, 2005) modified by the Ribeiro et al. Schinopsis brasiliensis Engl., Anacardiaceae, stem bark was (2009). Larvae were exposed to initially standard solutions collected in August 2011 in the city of Piranhas in Alagoas, of the HEE and its fractions (1000 μg/ml, in distilled water Brazil, whose geographical coordinates were identified by a containing 1% DMSO). That ismortalities exceeded 90%, with Garmin Forerunner GPS function (9°35’54.37” S and 37°46’08.31” further tests concentrations lower (100, 250, and 500 μg/ml) W). The species was identified by the biologist Marta Maria were performed to estimate the lethal concentration. 300 Clisiane C.S. Santos et al. / Rev Bras Farmacogn 24(2014): 298-303 Positive controls were treated with temephos [(O,O’-(thiodi- According to Bussmann et al. (2011), toxicity levels can be 4,1-phenylene) bis (O,O-dimethylphosphorothiolate)], defined the high toxicity (LC50 < 249 μg/ml), moderate toxicity (250 which was obtained as a commercial sample (3 μg/ml) from < LC50 < 499 μg/ml), low toxicity (500 < LC50 < 1000 μg/ml), and free
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