AFPP DIXIÈME CONFÉRENCE INTERNATIONALE SUR LES RAVAGEURS EN AGRICULTURE MONTPELLIER 22 ET 23 OCTOBRE 2014

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1 AFPP DIXIÈME CONFÉRENCE INTERNATIONALE SUR LES RAVAGEURS EN AGRICULTURE MONTPELLIER 22 ET 23 OCTOBRE 2014 COMPOSITION AND FUMIGANT TOXICITY OF ESSENTIAL OIL FROM ARTEMISIA HERBA-ALBA AGAINST TWO STORED PRODUCT BEETLES O. BACHROUCH 1, N. FERJANI 2, J. MEDIOUNI- BEN JEMÂA 3 1- Laboratoire de protection des végétaux, Institut National de la Recherche Agronomique de Tunisie 2- Institut Supérieur des Sciences Biologiques Appliquées de Tunis 3- Laboratoire de Biotechnologie Appliquée à l Agriculture, Institut National de la Recherche Agronomique de Tunisie RESUME Cette étude a porté sur l évaluation de la toxicité par fumigation de l huile essentielle de l armoise blanche Artemisia herba-alba en provenance de la Tunisie contre deux coléoptères des denrées stockées. Les analyses chromatographiques de l huile essentielle ont montré que le 1,8 cinéole (19.59%), le camphre (11.48%) et l α-pinène (5.4%) représentent les composés majeurs. Les tests de fumigation ont révélé que l huile essentielle est plus toxique contre Oryzaephilus surinamensis (L.) (LC 50 = µl/l air) que Tribolium castaneum (Herbst) (LC 50 = µl/l air). Mots-clés : huile essentielle, Artemisia herba-alba, Tribolium castaneum, Oryzaephilus surinamensis, CPG/SM, effet fumigène. ABSTRACT This study was conducted to investigate fumigant toxicity of Tunisian Artemisia herba-alba essential oil against two storage pest beetles. GC/MS analysis of essential oil showed that 1,8 cineol (19.59%), camphor (11.48%) and α-pinene (5.4%) were the main compounds in the oil. Fumigant test indicated that essential oil was more toxic against Oryzaephilus surinamensis (L.) (LC 50 = µl/l air) than Tribolium castaneum (Herbst) (LC 50 = µl/l air). Keywords: essential oil, Artemisia herba-alba, Tribolium castaneum, Oryzaephilus surinamensis, GC/MS, fumigant toxicity.

2 INTRODUCTION Insects are the main problem in stored grain as they affect their quantity and their quality (Madrid et al., 1990). The rust-red flour beetle Tribolium castaneum (Herbst, 1797) and the sawtoothed grain beetle Oryzaephilus surinamensis (L., 1758) are the most important and destructive stored product pests attacking grains in Tunisia and North Africa (Jarraya, 2003). Zettler and Arthur (2000) reported that fumigants play a crucial role against stored product pests because their ability to kill a broad spectrum of pests and their easy penetration into the commodities while leaving minimal residues. Fumigation with methyl bromide and phosphine is the most economical tool for managing stored insect pests in flour mills or food factories (Bell, 2000; Fields and white, 2002). However, they have been reported that these substances contribute to maintain food hygien but there are serious arguments about their negative effects on human health and enviroment (Kostyukovsky et al., 2002). Therefore, there is an urgent need to search an alternative strategy to control these pests. In this context, among methods used in Integrated Pest Management (IPM), plants and essential oils have been screened for their potential uses as alternative to chemical control. A number of studies have demonstrated the insecticidal proprerties of Artemisia species essential oils against a wild range of pests. The genus Artemisia belongs to the Asteracae family including more than 500 different species distributed through the world (Bora and Sharma, 2011) growing in arid and semi-arid climates. Plants of this genus are usually small herbs and shrubs. Sharifian et al. (2012) reported that A. herba alba may have ay have insecticidal or biological activity against insects against Tribolium castaneum (Herbst), Callosobruchus maculatus (F.) and Rhyzopertha dominica (F.). In Tunisia, Artemisia herba-alba under Arabic name of Shih is widly distributed from the north to the south of the country. These herbs have always been of a great botanical and pharmaceutical interest and are employed in folk medicine for healing different diseases (Bora and Sharma, 2011). This study was undertaken to evaluate composition and insecticidal activities of A. herba alba against T.castaneum and O. surinamensis. MATERIAL AND METHODS Insect rearing The rust-red floor beetle T. castaneum and the saw-toothed grain beetle O. surinamensis were reared on wheat flour. They were maintained in the dark in growth chambers sets 25 ± 1 C and 65% ± 5% (Relative Humidity). Adult insects, 7 days old, were used for fumigant toxicity tests. All experiments were carried out under the same environmental conditions. Plant material Artemesia herba-alba was collected from the region of Sawef (Zagouhan, North of Tunisia) in July The harvest material was air-dried at room temperature (20-25 C) for two weeks and stored in cloth bags.

3 Essential oil extraction and analysis Artemisia herba-alba essential oil was extracted from leaves (50 g of each sample in 500 ml of distilled water) submitted to hydrodistillation using a modified Clevenger type apparatus during 90 minutes. This time was fixed after a kinetic survey. The chemical composition was performed using GC-MS analysis. The confirmation of all components identification was achieved by calculating their kovats index. Fumigant toxicity bioassays Fumigant toxicity trials were carried out into Plexiglas bottles of 44 ml. Ten adult insects (7 days old) were placed in each bottle. Filter papers (2 cm diameter, whatman 1) were impregnated with appropriate doses of essential oil without using any solvent. The doses tested were 2.5 and 10µl corresponding respectively to concentration of and µl/l air. Each treatment and check was replicated four times. Mortality was recorded each 1 hour during 48h of exposure. When no leg or antennal movements were observed, insects were considered dead. Mortality was calculated using the Abbott (1925) formula. Lethal concentrations LC 50 and LC 95 A trial was designed to evaluate the LC 50 and LC 95. The following oil doses used were 2.5; 5; 7.5 and 10 µl. The number of dead and alive insects in each bottle was counted after 24 h of exposure and the LC 50 and LC 95 values.were estimated using Probit analysis (Finney, 1971). RESULTS Essential oil composition Analysis of A. herba-alba essential oil extracted during flowering stage from Sawef region showed that 22 compounds were identified which represent % of total constituents. This oil was mainly composed of α-pinene (5.40%), camphor (11.48%) and 1,8 cineol (19.59%). Fumigant toxicity Results showed that A. herba-alba essential oil was toxic to the two stored-beetles (Figure 1). The results led to that fumigant toxicity varied with insect species, oil concentration and exposure time. While for A. herba-alba essential oil, the lowest concentration (56.82 µl/l air) achieved 12% mortality of T. castaneum and 100% mortality of O. surinamensis after 48h exposure. However, at the highest concentration ( µl/l air) of A. herba-alba essential oil, 60% and 100% mortality was recorded respectively for T. castaneum and O. surinamensis. Figure 1: Percentage of mortality of T. castaneum and O. surinamensis exposed for various periods of time to Essential oil from A. herba-alba (Fumigant toxicity) Figure 1 : Pourcentage de mortalité de T.castaneum et O. surinamensis exposés à différentes durée à l huile essentielle d A. herba alba(fumigation) µl /l air µl/l air Exposure time (h) of T. castaneum Exposure time (h) of O. surinamensis

4 Furthermore, the corresponding LC 50 and LC 95 were respectively and µl/l air for T. castaneum against and µl/l air for A. herba-alba essential oil. DISCUSSION Twenty two compounds of A. herba-alba essential oil were identified using GC and GC/MS analysis from essential oil collected from Sawef (Zaghouan, North Tunisia). The main compounds were α-pinene (5.40%), camphor (11.48%) and 1,8 cineol (19.59%). The first investigation on A. herba-alba dates back to 1987 on isralean samples and revealed the existence of five different chemotypes (Segal et al., 1987). In fact, previous investigations by Feuerstein et al. (1988) showed that A. herba-alba essential oil was characterized by a high percentage of 1,8 cineol (13.3%) and Camphor (15%) followed by α-terpineol (6.3%) and Borneol (4.8%). In the same context, Bakkali et al. (2008) presented α-thujone, β-thujone (57%) and camphor (24%) as the dominating compounds of A. herbaalba essential oil. In addition, β-thujone, α- thujone, 1,8- cineole and camphor were the major components of Tunisian A. herba-alba essential oil (Mighri et al., 2010). In this work, Tunisian A. herba-alba essential oil exhibited strong fumigant toxicity against the rust-red flour beetle Tribolium castaneum (Herbst, 1797) and the saw-toothed grain beetle Oryzaephilus surinamensis (L., 1758). Results showed that A. herba alba essential oil from Tunisia was more toxic against Oryzaephilus surinamensis (L.) (LC 50 = µl/l air) than Tribolium castaneum (Herbst) (LC 50 = µl/l air). Our findings are in accordance with literature studies in which Artemisia vestia essential oil possessed strong fumigant toxicity against Sitophilus zeamais adults with a LC 50 value of mg/l air (Chu et al., 2010). In this context, half-life time of Artemisia scoparia essential oil for C. maculatus was longer than Sitophilus oryzae and Tribolium castaneum. These results showed the efficacy of this oil for use in organic food protection (Negahban et al., 2006). In addition, it has been reported that the essential oil of Artemisia monosperma obtained by steam distillation of the aerial parts of the plant was shown to have insecticidal activity against house fly, cotton leaf worm and the rice weevil (Saleh, 1984). Furthermore, Turi et al. (2014) indicated that Artemisia species had excellent potential applications for production of industrial chemicals, synergists and/or adjutants for use in agricultural applications. REFERENCES Abbott, W.S. (1925) - A method of computing the effectiveness of an insecticide. Journal of Economic Entomology. 18, Bakkali, F., Averbeck, S., Averbeck, D., Idaomar, M., Biological effects of essential oils. Food and Chemical Toxicology. 46, Bell, C.H Fumigation in the 21st century, Crop Protection 19, Bora, K.S., Sharma, A., The genus Artemisia: a comprehensive review. Pharmaceutical Biology. 49, Fields, P.G., White, N.D.G., Alternatives to methyl bromide treatments for stored-product and quarantine insects. Annual Review of Entomology 47, Finney. D.J., Probit analysis. Cambridge Univ. Press, Cambridge, 333 pp.

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