Toxicological Effect from the Stem Cortex of the Amazonic Plant Soapberry Paullinia Clavigera (Sapindaceae) Upon Three Arthropods
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Cien. Inv. Agr. 37(3):133-143. 2010 www.rcia.uc.cl RESEARCH NOTE Toxicological effect from the stem cortex of the amazonic plant soapberry Paullinia clavigera (Sapindaceae) upon three arthropods Diana Pérez1, José Iannacone2, and Harvey Pinedo1 1Instituto de Investigaciones de la Amazonía Peruana (IIAP), Ucayali, Jr. Progreso 102, Pucallpa, Ucayali, Perú. 2Universidad Nacional Federico Villarreal, Facultad de Ciencias Naturales y Matemática, Laboratorio de Ecofisiología Animal. Av. Río de Chepén, s/n Urb. Villa Hermosa, Bravo Chico, El Agustino, Lima, Perú. Abstract D. Pérez, J. Iannacone, and H. Pinedo. 2010. Toxicological effect from the stem cortex of the amazonic plant soapberry Paullinia clavigera (Sapindaceae) upon three arthropods. Cien. Inv. Agr. 37(3):133-143. Toxicological effects of four extracts proceeding from the stem cortex of the amazonic plant Paullinia clavigera D.R. Simpson (Sapindaceae) by decoction, ethanolic, chlorophormic and hexanic on three arthropods: Rhynchophorus palmarum (Linnaeus, 1758) (Curculionidae), Eupalamides cyparissias (Fabricius, 1777) (Castniidae) and Artemia franciscana (Kellog, 1906) (Artemiidae) in Ucayali, Peru were studied. The four extracts at the highest concentration tested were: decoction at a proportion 1:10 (w/v), ethanolic, chlorophormic and hexanic at 100 mg·L-1. Toxic effects were evaluated at 12, 24, 48 and 72 hours on larvae of III instar of R. palmarum and larvae of II instar of E. cyparissias; and at 24 and 48 hours on nauplii of A. franciscana. Toxicity in terms of LC50 on the three arthropods evaluated depends of type of extract of soapberry employed. In R. palmarum, decoction (LC50-72H = 59.15%) presented a high toxicity, although a significant effect of hydroalcoholic extract was observed at 40 mg·L1 in comparison with control. The extracts of decoction (LC50-72H = 70.71 %) and ethanolic (LC50-72H -1 = 66.21 mg·L ) presented high toxicities on E. cyparissias, and finally, hexanic extracts (LC50- -1 - 48H = 18.79 mg·L ), decoction (LC50-48H = 23.82 %) and chlorophormic (LC50-48H = 23.64 mg·L 1) presented the highest toxicities on A. franciscana. In the phytochemical analysis, saponins showed a very positive reaction in hydroalcoholic extract, and flavonoids and phenols had a very positive reaction in extract of decoction. The triterpenes were present only in the hexane extract. The hydroalcoholic and decoction extract showed toxicity on the two pest R. palmarum and E. cyparissias, but only the hydroalcoholic compared to the other three extracts had the lowest effect of risk and higher selectivity to the aquatic environment on A. franciscana. Key words: Artemia franciscana, botanical insecticide, ecotoxicology, Eupalamides cyparissias, Paullinia clavigera, Rhynchophorus palmarum. Introduction urgent to have alternative technologies in con- trol of plague insects and diminish the use of synthetic agrochemical products contaminat- Nowadays, products of low toxicity and low ing and impacting the environment negatively cost are required through the sustainable use (Iannacone et al., 2002a). The Ucayali region of natural resources; therefore, it becomes in the Peruvian Amazonia has a wide biodiver- sity of vegetal species that may be used as bio- Received August 17, 2009. Accepted December 16, 2009. cides (Pérez and Iannacone, 2006a; Iannacone Corresponding author: [email protected] et al., 2007). 5314 -12 PEREZ 37-3.indd 133 20-12-2010 16:16:50 134 CiEnCiA E inVEStigACión AgRARiA Paullinia clavigera Schldl. Var. Bullata D.R. for the evaluation of substances physiologically Simpson (Sapindaceae) “Soapberry”, is an en- bioactive and plaguicides whose destination is demic semi-woody vejuco (stem cortex), which the aquatic environment (Machera et al., 1996; is part of the primary forest and shores of South Hlywka et al., 1997; De los Ríos and gajardo, American Amazonic aquatic ecosystems (Fe- 2004, Gajardo et al., 2004, Cuadra et al., 2005). don and Castillo-Suarez, 2005; Jǿrgensen et al., 2005; Weckerle and Rutishauser, 2005; Bridge- The objectives of the present work were the de- water et al., 2006; Iannacone et al., 2007; Ur- termination of the toxicological effect of four dampilleta et al., 2007). This species has been extracts from the stem cortex of the Amazo- quoted as an ethnobiocide, as the root and the nic plant P. clavigera (Sapindaceae) obtained bark are used for artisanal fishing (Schultes and by aqueous, hydroalcoholic, chlorophormic Raffauf, 1990). Additionally, it presents anti- and hexanic decoction on three arthropods: fungal and molluscicide activity (Ekabo et al., R. palmarum (Curculionidae), E. cyparissius 1996). Toxicity has been related to the presence (Castniidae) and A. franciscana (Artemiidae) of saponines. On the other hand, the presence of in Ucayali, Peru and the characterization of the triterpens, B-sitosterol and ethereal oils is men- phytochemistry of the four extracts obtained. tioned, where the ictiotoxic activity is attributed to the triterpens (Texeira et al., 1984; Gupta, 1995; Iannacone et al., 2007). A bark and root Materials and methods infusion is used for washing mycosis and skin wounds. It may be also taken orally after mac- Obtaining botanic extracts eration in hard liquor (Vela, 2003). Four types of extracts of P. clavigera were used: Eupalamides cyparissias (Fabricius, 1777) aqueous extract by decoction, hydroalcoholic, (Lepidóptera: Castniidae) is commonly known hexanic and chlorophormic. The preparation as “bunch and stipe borer” of palm trees (Schu- of the different types of extracts of P. clavig- iling and Van Dinther, 1980). Damage is made era were maintained in conditions of diffused by the larvae instars affecting inflorescences or light with 32 watts t8 fluorescent tubes, at 25.3 flowering bunches resulting in reduced size and ± 2 ºC of temperature and at 77.6 ± 3% of rela- weight. When the formed or mature bunches are tive humidity. The material of P. clavigera was affected, it provokes the total or partial rot of obtained from the gens bank of medicine plants fruits (Pérez and Iannacone, 2006b, 2008). and biocides in the IIAP Ucayali branch, located in the Km 12,400 of the Federico Basadre high- Rhynchophorus palmarum (Linnaeus, 1758) way, Pucallpa, Ucayali, Peru. The vejucos (stem (Coleoptera: Curculionidae), “palm weevil” cortex) were dried under direct sunlight, after (Mexzón, 1994), causes damage by the instars their segmentation. Then, the dry vejucos were larvae in the pejibaye plant, attacking the plant ground with a hammer mill. A 1:10 proportion rhizome. In other palm trees, it forms passage- was used for the preparation of the aqueous ex- ways in the trunk, leaves and petioles (Vela, tract by decoction. The preparation was boiled 2003; Pérez and Iannacone, 2006c). for 2 hours in an electric stove, until 1L of con- centrated solution was obtained (250 g·L-1). It Artemia franciscana (Kellog, 1906) (Anos- was filtered with polysilk meshes® (Polyester, traca: Artemiidae) is known as “brine shrimp”. Melilla, Spain), to separate the extract of the It is an aquatic species widely used in ecotoxi- residues. It was cooled for 2 hours. In order to cological tests as non-target organism, as it is evaluate the two insects from the concentrated an easily obtainable biological material with solution, concentrations of the aqueous extracts wide geographic distribution, good cultivation by decoction were prepared, denominated at feasibility and inexpensive to obtain. The bi- 100% (= 100 g·L-1), 75% (= 75 g·L-1), 50% (=50 otests with this species are easily to evaluate, g·L-1) and 25% (= 25 g·L-1). Distilled water at manipulate and read (Iannacone et al., 2002b). pH 7 was used as control. The preparation of It is used in biotests of phytochemical research the hexanic, chlorophormic and hydroalcoholic 5314 -12 PEREZ 37-3.indd 134 20-12-2010 16:16:50 VOLUME 37 nº3 SEPtEMBER - DECEMBER 2010 135 extracts were made from 100 g of a dry and positive. + = reaction slightly positive. ++ = grounded sample in Laboratorio de Produc- positive reaction. +++ = reaction very positive tos Naturales Antiparasitarios de la Amazonia (Lock, 1994) (UNAP), Iquitos, Peru. They were extracted with hexane (apolar solvent) until they were ex- hausted with renovation solvent every 48 hours; Biological material the hexanic extracts were obtained after the sol- Rhynchophorus palmarum. Larvae of the VI to vent was eliminated in rotary evaporator under the IX instars were obtained from the Central reduced pressure. Chlorophorm (CHCl , solvent 3 Market of Pucallpa, Ucayali, Peru. The larvae moderately polar) was added to the residue until were arranged individually in 1-liter-containers it was exhausted with solvent renovation every and they were fed with pieces of Bactris gasi- 48 hours, the solvent was eliminated to dry- paes Kunth (palm heart) (3.5 g·day-1). Four lar- ness in rotary evaporator under reduced pres- vae were placed with food in 25 x 15 x 10 cm- sure, therefore, the chlorophormic extracts were plastic-containers. The adults obtained were obtained. Finally, ethanol-water (70:30) (polar placed in couples in 1-liter-containers for mat- solvents) was added to the residue and the hy- ing. Distilled water was sprayed to the layings droalcoholic extracts were obtained, as in the to maintain humidity. The eclosioned larvae previous cases. were placed individually in 5 x 5 x 3 cm con- tainers and they were fed with palm heart (1.5 g), 100 mg of the heavy extract was diluted using which was renewed every two days, until reach- 0.5 mL of hexane, 0.75 mL of chlorophorm and ing the III instar, which occurs eight days after 0.45 mL of ethylic alcohol as dissolvent, accord- the larvae eclosion (Bartra, 1994). ing to the extract. The amounts of the dissolvent used in each case were different depending on Eupalamides cyparissias. Adult egg-filled fe- the minimum mL for the extracts dissolution. males were collected in Km 60 of the Federico Then, they were diluted in 1 L of distilled water Basadre highway, neshuya highway– Curi- adjusted to pH of 7.