Boletín Latinoamericano y del Caribe de Plantas Medicinales y Aromáticas ISSN: 0717-7917 [email protected] Universidad de Santiago de Chile Chile

CABALLERO-GALLARDO, Karina; OLIVERO-VERBEL, Jesús; PINO-BENÍTEZ, Nayive; STASHENKO, Elena E. Chemical composition and bioactivity of auritum and P. multiplinervium essential oils against the red flour beetle, Tribolium castaneum (Herbst) Boletín Latinoamericano y del Caribe de Plantas Medicinales y Aromáticas, vol. 13, núm. 1, 2014, pp. 10-19 Universidad de Santiago de Chile Santiago, Chile

Available in: http://www.redalyc.org/articulo.oa?id=85629766002

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Caballero-Gallardo et al. Chemistry of Piper oils and bioactivity against Tribolium castaneum

INTRODUCTION these chemicals in hot spots for biodiversity. Colombia In a world where the use of synthetic chemicals is is a country with approximately 45.000 plant species, common in various areas of human life, and concern and it is among the first countries throughout Latin for sustainable product development has increased, America and the world in terms of biodiversity natural resources emerge as a source of raw material heritage. However, little is known about its richened solution for effective and environmentally friendly and the enormous possibilities for innovation and products. Nowadays, a number of synthetic sustainable development from this natural resource insecticides and fumigants are used to protect grain (Stashenko et al., 2010). At the same time, as people storage. Infestation of storage grains is a very serious depend on synthetic repellents to control pests of problem as various life stages of insects cause stored products, EOs are exported without any added economic damage and deteriorates the quality of food value, reducing the income for many families, and the grains and food products. These pests often cause ability to create jobs through the manufacture of extensive loss of products stored in tropical and products derived from these natural resources. semitropical environments (Isman, 2000). Several The bioactivity of a large number of essential species of insect pests are known for attacking oils and their constituents have been evaluated against granaries and other food products since ancestral T. castaneum (Jahromi et al., 2012; Amin et al., 2012; times. Tribolium castaneum is major pest food Zandi-Sohani et al., 2013; Liang et al., 2013). In products (Mishra and Tripathi, 2011) the adult stage of Colombia, EOs from Cananga odorata, Lippia alba, T. castaneum is very active and can breed throughout L. orignanoides, citratus, C. martinii, C. the year in warm areas (Pugazhvendon et al., 2009). flexuosus, citriodora, and Citrus sinensis They live two years or more, during which female have been tested as repellents using this insect produce nearly 1000 eggs (Shukla et al., 2010). (Olivero-Verbel et al., 2009, 2010; Caballero-Gallardo However, the use of synthetic pesticide for the et al., 2011, 2012). However, species from the management of these insect pests cause great hazards Piperaceae family have not been studied. In this for environmental and toxic to non target livings context, the aim of this paper was to evaluate the (Mishra and Tripathi, 2011). repellent and insecticidal properties of EOs isolated Over the years, pesticide use has led to a from Piper auritum and P. multiplinervium cultivated variety of problems such as environmental pollution, in Chocó, Colombia against T. castaneum Herbst, one chemical residues in food grains, development of of the most far-flung and destructive stored-product insecticide resistance and toxicity to non target pests throughout the world (Zapata and Smagghe, organisms (Cosimi et al., 2009; Sousa et al., 2009). 2010). The increasing public awareness about pesticide safety and possible damage to the environment has resulted MATERIALS AND METHODS in greater attention being given to natural products for Plant Material the control of food stored pests (Rajendran and Two plant species of the genus Piper were used in the Sriranjini, 2008). Among the group of natural assays, P. auritum and P. multiplinervium (Figure 1). molecules used to control insects, essential oils (EOs) Plant material was collected from Chocó, Colombia, at area being considered as outstanding solutions for the municipalities of Quibdó and Lloró, respectively). their effectiveness and versatility. In fact, volatility The taxonomic identification of the plants was carried and chemical diversity makes them excellent out in the Colombian National Herbarium (COL) of repellents. The use EOs derived from aromatic plants the Institute of Natural Sciences, Faculty of Sciences as low-risk insecticides has increased considerably of the National University of Colombia (Bogotá) and owing to their popularity with organic farmers and stored with voucher specimens COL No. 512209 and environmentally conscious consumers (Regnault- COL No. 519977 for Piper auritum and P. Roger et al., 2012). multiplinervium, respectively. EOs have repellent, insecticidal, and growth- reducing effects on a variety of insects (Rajendran and Sriranjini, 2008; Ukeh et al., 2009; Mondal and Khalequzzaman, 2010). The demand for naturally active compounds such as EOs has move forward the necessity to search Boletín Latinoamericano y del Caribe de Plantas Medicinales y Aromáticas/11

Caballero-Gallardo et al. Chemistry of Piper oils and bioactivity against Tribolium castaneum

Figure 1

Studied species from Chocó, Colombia.

Extraction of essential oils were based on Kováts indices (Ik) and by comparison Essential oils were extracted from fresh leaves by of the mass spectra fragmentation patterns with those microwave-assisted hydrodistillation (MWHD) as found in databases or libraries (NIST02, Adams, previously described by Stashenko et al. (2004). The Wiley7n ) (Adam, 2004). process used a Clevenger type hydrodistillation equipment placed inside a domestic microwave LG Insects and rearing conditions microwave MS- 1242 ZK model, set at 2450 MHz, All experiments were conducted in the laboratory 800 W. The extraction was conducted in 10 intervals; using long-established colonies of T. castaneum. The 10 min for each one with 5 min rest time each one. strain was kept in glass containers covered with a The oil was dried using anhydrous sodium sulphate. plastic mesh. Insects were reared on a diet of whole oat flour and kept at 26 ± 2 °C, with a relative Gas chromatography–mass spectrometry (GC–MS) humidity of 70 - 85%, and a 10:14 h light:dark An aliquot of each essential oil (20 μL) was dissolved photoperiod. Red flour beetle adults of both sexes in dichloromethane to a final volume of 1 mL, and were collected, and those with size between 3.0 and then transferred to vials for GC–MS. The 4.5 mm, and 7-10 days old, were employed in the chromatographic analysis was performed using an experiments (Caballero-Gallardo et al., 2011). Agilent Technologies 6890 Plus (Palo Alto, CA) GC coupled to an Agilent Technologies MSD 5975 Repellency tests selective detector mass equipped with a split/splitless The repellent activity was measured using the area injector port (1:50 split ratio), an automatic injector preference method (Tapondjou et al., 2005; Olivero et Agilent 7863, and a data system HP ChemStation. One al., 2009; Olivero-Verbel et al., 2010; Caballero- microliter of solvent was injected into the GC–MS Gallardo et al., 2011; Lü and Shi, 2012). The solutions equipment for the corresponding chromatographic of EOs were prepared in acetone, and in all cases, a analysis. A 60m×0.25mm i.d.×0.25μm with 5% volume of 0.5 mL was evenly applied to a half-filter phenyl poly(methylsiloxane) stationary phase. The paper disk to obtain the desired oil volume per area oven temperature was set at 45 °C for 5 min, then unit of 0.00002, 0.0002, 0.002, 0.02, and 0.2 μL/cm2. increased 4 °C/min up to 150°C for 2 min, then to 5 The other half of the filter paper was treated with an °C/min up to 250 °C for 5 min, and finally at equal volume of acetone as a vehicle control. Test 10°C/min up to 275°C. Helium was used as a carrier areas consisted of 9 cm Albet DP 597125 filter paper gas with 16.47 psi column head pressure and 1 cut in half (31.8 cm2). A formulation of IR3535 [ethyl mL/min linear velocity. The components identification 3-(N-acetyl-N-butylamino) propionate], a synthetic

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Caballero-Gallardo et al. Chemistry of Piper oils and bioactivity against Tribolium castaneum

repellent (Licciardi et al., 2006; Faulde et al., 2010) respectively. Comparisons between mean PRs for which has been approved in the USA for skin evaluated EOs were obtained using ANOVA, with applications (World Health Organization, 2001), was Dunn’s post-test used to compare treated with control- employed as a positive control, utilizing the same vehicle groups. Statistical analysis was performed with experimental conditions as the oils. The treated and GraphPad 3.00. The mean repellence concentration control half disks were air-dried for 10 min to remove (RC50) values were calculated using probit analysis the solvent, re-attached with adhesive tape, and kept in (Finney, 1971). For all purposes, significance was set 90 mm glass Petri dishes. Twenty adults of T. at P < 0.05. castaneum of both sexes were released at the center of each filter paper disk. Dishes were covered and placed RESULTS in darkness at 26 ± 2 °C and relative humidity of 70– The results of repellency assays for tested EOs are 85%. The numbers of T. castaneum specimens on presented in Table 1. Data showed that at tested treated and untreated portions of the experimental concentrations, both EOs were strongly repellent paper halves were counted for each dish after 2 and 4 against T. castaneum. At the lowest assayed h exposure. Percentage repellency (PR) for a given concentration (0.00002 μL/cm2), the EOs isolated treatment time was obtained using the formula: from P. auritum and P. multiplinervium showed attractant activity at both exposure times (PR: -35 ± PR = [(Nc-Nt)/(Nc+Nt)] x 100 9%, -30 ± 17% and -5 ± 10%, -8 ± 13%, respectively). The commercial repellent IR3535 was less effective Where Nc and Nt are the number of insects on the than the two tested EOs, when the organisms were untreated (control) and treated areas, respectively. exposed to 0.02 and 0.2 μL/cm2. Three replicates were used for each tested Significant differences were found for the PR concentration of EO, and each assay was repeated between the essential oil of P. auritum and the twice. commercial repellent (IR3535) at higher concentrations (0.02 and 0.2 μL/cm2). However, for Contact toxicity on filter papers the essential oil isolated from P. multiplinervium, no The contact toxicity on filter papers was conducted such differences were recorded. using filter paper discs (Albet DP 597125, 9 cm RC50 values are shown in Table 2. Repellent diameter) (Tapondjou et al., 2005). Oils were action was highly dependent upon oil concentration dissolved in acetone at concentrations of 0.2, 0.5, 0.9 and exposure time. P. auritum essential oil was the 2 2 and 1.2 μL/cm , and 1 mL of each solution was most active with a RC50 value of 0.002 μL/cm . uniformly distributed on the surface of the paper that The results for the contact lethality on filter was then placed in glass Petri dishes. After 10 min, paper for the examined EOs are shown in Table 3. once the solvent had been evaporated, 20 unsexed The EO from P. auritum had greater toxicity adults were transferred into each disc and stored in than that from P. multiplinervium. After 24 h darkness at 26 ± 2 °C and 75 ± 10 % RH (Olivero et exposure, the first EO caused 81±2% mortality at the al., 2009; Olivero-Verbel et al., 2010; Caballero- lowest tested concentration (0.2 μL/cm2), but at 0.9 Gallardo et al., 2011). Three replicates were employed μL/cm2 196 this was 100%. However, for P. for each treatment, repeating each assay twice. multiplinervium, the maximum tested concentration Mortality was recorded after 24, 48 and 72 h. IR3535 (1.2 μL/cm2) and exposure time (72 h) only produced was utilized as a positive control. Insects were 16±4% lethality. considered dead when no leg or antennal movements The results obtained to repellent and mortality were recorded. of essential oils against T. castaneum were analyzed using Student's t-test. In the case of the repellent Data analysis activity were no significant differences were observed The paired t-test was utilized to compare mean number between the two essential oils, whereas significant of insects on the treated and untreated area of the filter differences were observed to essential oil P. auritum paper. Repellency or attractancy was established if (Table 3). significant differences occurred for positive or The IR3535 presented very little effect on the negative percentage repellency, respectively. Normal survival of T. castaneum adults. Chemical composition distribution and equality between variances were of EO isolated from the two Piper species, as resulted checked by Kolmogorov-Smirnov and Bartlett’s tests, from GC-MS analysis, is presented in Table 4. The Boletín Latinoamericano y del Caribe de Plantas Medicinales y Aromáticas/13

Caballero-Gallardo et al. Chemistry of Piper oils and bioactivity against Tribolium castaneum

results revealed quite different compositions between composition, whereas P. auritum is mainly composed the two species. The essential oil of P. auritum was by safrole (93.2%). This chemically based difference characterized by high phenylpropanoid content may explain the acute and low-dose toxicity observed (97.5%), in which safrole (93.2%) was found to be the for P. auritum (100% mortality after 24 h exposure, 2 major constituent and miristicine the second (4.3%). and RC50= 0.002 μL/cm ). Among monoterpene hydrocarbons, -terpinene was The bioactivity of the EOs results from found at low concentration (0.3%). The P. interaction among structural components, particularly multiplinervium oil presented a high content of the major constituents, although, the other compounds sesquiterpene hydrocarbons (67%) with β-elemene in the oil may also have a vital function (Intirach et al., (8.7%), trans-β-caryophyllene (5.3%) α-selinene 2012). In this study the principal compound of P. (3.8%) and β-selinene (3.4%) as major components, as auritum was safrole (93.2%), a methylenedioxy well as oxygenated sesquiterpenes (7.1%), with compound (Maul et al., 2011), reported to have toxic caryophyllene oxide (4.1%) and trans-nerolidol (3.0%) properties derived from its structure as benzene as representative compounds. derivative (Tripathi et al., 2009). The metabolism of this naturally occurring chemical leads to the DISCUSSION formation of safrole 2',3'-oxide, an electrophilic Different plant products, especially EOs, have compound that produces DNA adducts in vitro and in considerable potential as insecticides and repellents, vivo (Shen et al., 2012), probably explaining its and they are gaining tremendous importance for the reported ability to induce liver tumors in mice and rats management of stored products, as these mixtures are (Rietjens et al., 2005). Several studies have been considered biodegradable and ecologically safe. The published about safrole being the main component in red flour beetle, T. castaneum, is one of the most several Piper species (Abreu et al., 2002; Protti et al., widespread and pernicious destructive stored-product 2005; Sauter et al., 2012; Souto et al., 2012), and used pests throughout the world (Zapata et al., 2010; El- alone, it has been reported to be effective against T. Sayed and Genan, 2012). Results presented here have castaneum adults (Huang et al., 1999). shown that essential oils obtained from Piper auritum This work has clearly revealed that Piper and P. multiplinervium possess good bioactivity chemotypes can vary considerably in their toward this insect. In addition, these oils have similar composition and biological properties; and therefore, or better repellent properties than the commercial their presence in Colombian flora is an important repellent used as a positive control (IR3535). potential for promissory bioactive molecules, in Properties of EOs are directly related to their particular as repellents, as it has been also chemical composition (Ben Marzoug et al., 2011), that demonstrated for P. aduncum, a species from the in turn, are a function of the plant part extracted, time Brazilian biodiversity that has been shown to work as of collection and growth environment conditions, a repellent against the ant Solenopsis saevissima among others (Zapata and Smagghe, 2010). The (Smith) (Hymenoptera: Formicidae) (Souto et al., number and concentration of chemicals present in an 2012). EO may vary dramatically, even within the same In short, reported data revealed, for the first species (Zapata and Smagghe, 2010). In this study, P. time, the repellent and insecticidal activity of essential multiplinervium and P. auritum had different oils extracted from P. auritum and P. multiplinervium biological activities, reflecting their distinct chemical grown in the department of Chocó, Colombia, and this nature. P. multiplinervium does not contain a major property could be related to the presence of some component representing at least 50% of the oil particular compounds.

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Caballero-Gallardo et al. Chemistry of Piper oils and bioactivity against Tribolium castaneum

Table 1

Time 2 Hours 4 Hours Essential oils Concentration (µL/cm2) Concentration (µL/cm2)

0.00002 0.0002 0.002 0.02 0.2 0.00002 0.0002 0.002 0.02 0.2 P. auritum -35 ± 9ab 30 ± 7a 32 ± 11a 82 ± 3ab 90 ± 4ab -30 ± 17 38 ± 10a 45 ± 9a 55 ± 12a 87 ± 3a P. multiplinervium -5 ± 10 7 ± 13 42 ± 11a 70 ± 8a 82 ± 6a -8 ± 13 10 ± 16 45 ± 11a 70 ± 9a 80 ± 4a IR3535c 7 ± 13 17 ± 9 37 ± 15 50 ± 4a 72 ± 4a 5 ± 14 12 ± 8 42 ± 15a 50 ± 7a 72 ± 6a Percentage Repellency1 (PR) after two exposure times for two essential oils against T. castaneum 1Values are mean (SE of six replicates). aSignificant difference between the number of the organisms on both the treated and untreated halves, using a paired t test (P < 0.05). bSignificant difference between essential oils and the positive control (IR3535), using ANOVA, with Dunn’s post-test. c. Commercial repellent.

Table 2 RC50 (95% Confidence intervals) values of the essential oils tested against T. castaneum Exposure time RC Regression parameters* Essential oils 50 (h) (µL/cm2) R P 2 0.002 (0.001-0.005) 0.940 0.0602 P. auritum 4 0.002 (0.001-0.008) 0.920 0.0798 P. multiplinervium 2 0.008 (0.004-0.016) 0.967 0.0334 4 0.007 (0.003-0.015) 0.966 0.0336 IR3535a 2 0.015 (0.005-0.048) 0.997 0.0002 4 0.016 (0.006-0.045) 0.983 0.0027 *. From probit analysis. a. Commercial repellent.

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Caballero-Gallardo et al. Chemistry of Piper oils and bioactivity against Tribolium castaneum

Table 3 Concentration Mortality (%) Essential oils (µL/cm2) 24 h 48 h 72 h 0 0 ± 0 0 ± 0 0 ± 0 0.2 81 ± 2* 90 ± 4* 96 ± 2* Piper auritum 0.5 92 ± 2* 98 ± 1* 99 ± 1* 0.9 100 ± 0* 100 ± 0* 100 ± 0* 1.2 100 ± 0* 100 ± 0* 100 ± 0* 0 0 ± 0 0 ± 0 0 ± 0 0.2 0 ± 0 3 ± 2 9 ± 4 Piper multiplinervium 0.5 4 ± 2 7 ± 2 12 ± 1 0.9 5 ± 2 8 ± 2 15 ± 5 1.2 6 ± 2 10 ± 3 16 ± 4 0 0 ± 0 0 ± 0 0 ± 0 0.2 0 ± 0 0 ± 0 2 ± 1 IR3535* 0.5 0 ± 0 0 ± 0 3 ± 1 0.9 0 ± 0 1 ± 1 3 ± 1 1.2 1 ± 1 2 ± 1 4 ± 2

Mortality rates in T. castaneum exposed to EOs and IR3535 *. Significant difference between the two EOs, using Student's t-test (P < 0.05). a. Commercial repellent.

Table 4

Relative Composition (%) KIDB5a Compound Chemical Class Piper auritum Piper multiplinervium Monoterpene 1065 terpinene 0.3 hydrocarbons 1290 Safrole 93.2 Phenylpropanoids 1520 Miristicine 4.3 1398 β-elemene 8.7 1420 trans-β-caryophyllene Sesquiterpene 5.3 1490 β- selinene hydrocarbons 3.4 1497 α- selinene 3.8 1566 trans-nerolidol Oxygenated 3.0 1584 Caryophyllene oxide sesquiterpenes 4.1

Chemical composition (%) of tested essential oils aKovats index on DB-5 column

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Caballero-Gallardo et al. Chemistry of Piper oils and bioactivity against Tribolium castaneum

ACKNOWLEDGEMENTS essential oils from Mediterranean plants This work was supported by the University of against stored-product pests: Evaluation of Cartagena, Cartagena; CENIVAM, Bucaramanga; and repellency against Sitophilus zeamais Colciencias, Bogotá, Colombia. Grant RC-245-2011 Motschulsky, Cryptolestes ferrugineus Colciencias, Patrimonio Autónomo del Fondo (Stephens) and Tenebrio molitor (L.). J Nacional de Financiamiento para la Ciencia, la Stored Prod Res 45: 125 - 132. Tecnología y la Innovación, Francisco José de Caldas, El-Sayed HS, Genan MG. 2012. Food consumption and the National Program for Doctoral Formation and utilization by Tribolium confusum du Val (COLCIENCIAS, 567-2012). (Coleoptera:Tenebrionidae) larvae and their susceptibility to the acetone extract of Nerium REFERENCES oleander L. (Apocynaceae) leaves in relation Abreu AM, Sevegnani L, Machicado AR, Zimermann to three types of flour. J Stored Prod Res 51: D, Rebelo RA. 2002. Piper mikanianum 56 - 60. (Kunth) steudel from Santa Catarina, Brazil-a Faulde MK, Albiez G, Nehring O. 2010. Insecticidal, new source of safrole. J Essent Oil Res 14: acaricidal and repellent effects of DEET- and 361 - 363. IR3535-impregnated bed nets using a novel Adams RP. 2004. Identification of essential oil long-lasting polymer-coating technique. components by gas chromatography/qua- Parasitol Res 106: 957 - 965. drupole mass spectrometry, 4th ed., Allured Finney DJ. 1971. Statistical Method in Biological Publishing Corporation. Carol Stream, Illinois, Assay, 2nd edn. Griffin, London, UK. USA. Intirach J, Junkum A, Tuetun B, Choochote W, Amin R, Mondol R, Rahman F, Alam J, Habib Chaithong U, Jitpakdi A, Riyong D, R, Hossain T. 2012. Evaluation of insecticidal Champakaew D, Pitasawat B. 2012. Chemical activity of three plant extracts against adult constituents and combined larvicidal effects of Tribolium castaneum (Herbst). Biologija 58: selected essential oils against Anopheles 37 - 41. cracens (Diptera: Culicidae). Psyche 2012: 1 - Ben Marzoug HN, Romdhane M, Lebrihi A, Mathieu 11. F, Couderc F, Abderraba M, Khouja M. L, Isman MB. 2000. Plant essential oils for pest and Bouajila J. 2011. Eucalyptus oleosa essential disease management. Crop Prot 19: 603 - oils: chemical composition and antimicrobial 608. and antioxidant activities of the oils from Jahromi MG, Pourmirza AA, Safaralizadeh MH. 2012. different plant parts (stems, leaves, flowers Repellent effect of sirinol ( emulsion) and fruits). Molecules 16: 1695 - 1709. against Lasioderma serricorne (Coleoptera: Caballero-Gallardo K, Olivero-Verbel J, Stashenko E. Anobiidae) and Tribolium castaneum 2011. Repellent activity of essential oils and (Coleoptera: Tenebrionidae) by three some of their individual constituents against laboratory methods. African J Biotech 11: Tribolium castaneum Herbst. J Agric Food 280 - 288. Chem 59: 1690 - 1696. Liang Y , Li YL, Xu S, Zhao NN, Zhou L, Cheng J, Huang Y, Ho SH, Kini RM. 1999. Bioactivities of Liu ZL. 2013. Evaluation of repellency of Safrole and Isosafrole on Sitophilus zeamais some chinese medicinal essential oils (Coleoptera: Curculionidae) and Tribolium against Liposcelis bostrychophila (Psocoptera: castaneum (Coleoptera: Tenebrionidae). J Liposcelidae) and Tribolium castaneum Econ Entomol 92: 676 - 683. (Coleoptera: Tenebrionidae). J Econ Entomol Caballero-Gallardo K, Olivero-Verbel J, Stashenko E. 106: 513 - 519. 2012. Repellency and toxicity of essential oils Licciardi S, Herve JP, Darriet F, Hougard JM, Corbel from Cymbopogon martinii, Cymbopogon V. 2006. Lethal and behavioural effects of flexuosus and Lippia origanoides cultivated in three synthetic repellents (DEET, IR3535 and Colombia against Tribolium castaneum. J KBR 3023) on Aedes aegypti mosquitoes in Stored Prod Res 50: 62 – 65. laboratory assays. Med Vet Entomol 20: 288 Cosimi S, Rossi E, Cioni PL, Canale A. 2009. - 293. Bioactivity and qualitative analysis of some Boletín Latinoamericano y del Caribe de Plantas Medicinales y Aromáticas/17

Caballero-Gallardo et al. Chemistry of Piper oils and bioactivity against Tribolium castaneum

Lü JH, Shi YL. 2012. The Bioactivitiy of essential oil Sauter IP, Rossa GE, Lucas AM, Cibulski SP, Roehe from Ailanthus altissima swingle (Sapindales: PM, Alves da Silva L, Brittes Rott M, 321 Simaroubaceae) bark on Lasioderma Figueiro Vargas RM, Cassel E, von Poser G. serricorne (Fabricius) (Coleoptera: 2012. Chemical composition and amoebicidal Anobiidae). Adv Mat Res 365: 428 - 432. activity of Piper hispidinervum (Piperaceae) Maul R, Siegl D, Kulling SE. 2011. Glucuronidation essential oil. Ind Crops Prod 40: 292 - 295. of the red clover isoflavone irilone by liver Shen LC, Chiang SY, Lin MH, Chung WS, Wu KY. microsomes from different species and human 2012. In vivo formation of N7-guanine DNA UDP-glucuronosyl transferases. Drug Metab adduct by safrole 2',3'-oxide in mice. Toxicol Dispos 39: 610 - 616. Lett 18: 309 - 315. Mishra BB, Tripathi SP. 2011. Repellent activity of Shukla GS, Upadhyay VB, Mathur R, Prasad SG. plant derived essential oils against Sitophilous 2010. Economic zoology, biostatistics and oryzae (Linnaeus) and Tribolium castaneum animal behaviour. Rustogi Publication, India, (Herbst). Singapore. J Sci Res 1: 173 - 178. pp: 97. Mondal M, Khalequzzaman M. 2010. Toxicity of Sousa A, Faroni L, Pimentel M, Guedes R. 2009. naturally occurring compounds of plant Developmental and population growth rates of essential oil against Tribolium castaneum phosphine-resistant and susceptible (Herbst). J Biol Sci 10: 10 - 17. populations of stored product insect pests. J Olivero J, Caballero K, Jaramillo B, Stashenko E. Stored Prod Res 45: 241 - 246. 2009. Actividad repelente de los aceites Souto RNP, Harada AY, Andrade EHA, Maia JGS. esenciales de Lippia origanoides, Citrus 2012. Insecticidal activity of piper essential sinensis, y Cymbopogon nardus cultivadas en oils from the amazon against the fire Ant Colombia frente a Tribolium castaneum, Solenopsis saevissima (Smith) (Hymenoptera: Herbst. Salud UIS 41: 244 - 250. Formicidae). Pest Manag 41: 510 - 517. Olivero-Verbel J, Nerio L, Stashenko E. 2010. Stashenko E, Jaramillo B, Martinez J. 2004. Bioactivity against Tribolium castaneum Comparison of different extraction methods Herbst (Coleoptera: Tenebrionidae) of for the analysis of volatile secondary Cymbopogon citratus and Eucalyptus metabolites of Lippia alba (Mill.) N.E. Brown, citriodora essential oils grown in Colombia. grown in Colombia, and evaluation of its in Pest Manag Sci 66: 664 - 668. vitro antioxidant activity. J Chromatogr A Protti S, Fagnoni M, Albini A. 2005. Expeditious 1025: 93 - 103. synthesis of bioactive allylphenol constituents Stashenko E, Martinez J, Ruiz C, Arias G, Duran C, of the genus Piper through a metal-free Salgar W, Cala M. 2010. Lippia origanoides photoallylation procedure. Org Biomol Chem chemo type differentiation based on essential 3: 2868 - 2871. oil GC-MS and principal component analysis. Pugazhvendon SR, Elumalai K, Ross PR, J Separ Sci 33: 93 - 103. Soundararajan M. 2009. Repellent activity of Tapondjou AL, Adler C, Fontem DA, Bouda H, chosen plant species against Tribolium Reichmuth C. 2005. Bioactivities of cymol castaneum. World J Zool 4: 188 - 190. and essential oils of Cupressus sempervirens Rajendran S, Sriranjini V. 2008. Plant products as and Eucalyptus saligna against Sitophilus fumigants for stored-product insect control. J zeamais Motschulsky and Tribolium confusum Stored Prod Res 44: 126 - 135. du Val. J Stored Prod Res 41: 91-102. Regnault-Roger C, Vincent C, Arnason JT. 2012. Tripathi AK, Upadhyay S, Bhuiyan M, Bhattacharya Essential oils in insect control: Low-risk PR. 2009. A review on prospects of essential products in a high-stakes world. Annu Rev oils as biopesticide in insect-pest management. Entomol 57: 405 - 424. J Pharm Pharm 1: 052 - 063. Rietjens IM, Boersma MG, van der Woude H, Ukeh DA, Birkett MA, Pickett JA, Bowman AS, Jeurissen SM, Schutte ME, Alink GM. 2005. Luntz AJM. 2009. Repellent activity of Flavonoids and alkenylbenzenes: mechanisms , , and of mutagenic action and carcinogenic risk. , Zingiber officinale, against the maize Mutat Res 574: 124 - 138. weevil, Sitophilus zeamais. J Phytochem 70: 751 - 758. Boletín Latinoamericano y del Caribe de Plantas Medicinales y Aromáticas/18

Caballero-Gallardo et al. Chemistry of Piper oils and bioactivity against Tribolium castaneum

World Health Organization. 2001. Report of the Fourth WHOPES Working Group Meeting. WHO/CDS/WHOPES/2001.2. WHO, Geneva, Swizerland. Zandi-Sohani N, Hojjati M, Carbonell-Barrachina AA. 2013. Insecticidal and repellent activities of the essential oil of Callistemon citrinus (Myrtaceae) against Callosobruchus maculatus (F.) (Coleoptera: Bruchidae). Neotrop Entomol 42: 89 - 94. Zapata N, Lognay G, Smagghe G. 2010. Bioactivity of essential oils from leaves and bark of Laurelia sempervirens and Drimys winteri against Acyrthosiphon pisum. Pest Manag Sci 66: 1324 - 1331. Zapata N, Smagghe G. 2010. Repellency and toxicity of essential oils from the leaves and bark of Laurelia sempervirens and Drimys winteri against Tribolium castaneum. Ind Crop Prod 32: 405 - 410.

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