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The document discusses essential oils as potential insecticidal agents for protecting stored products. It provides background on stored product pests and current control methods using synthetic pesticides. Essential oils from various plant species have been screened for insecticidal activity against stored product insects. Studies found that essential oils from Chrysanthemum coronarium and Salvia veneris showed high insecticidal activity against Sitophilus granarius in screening tests. Further research is needed to develop essential oils as natural alternatives to synthetic pesticides for stored product protection.

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1 Essential oils as potential insecticidal agents for protection of stored products Kaan Polatoğlu1, Hüseyin Servi2, Ömer Cem Karakoç3,Yasemin Yücel Yücel4, Salih Gücel5, Betül Demirci6, Kemal Hüsnü Can Başer6,7 1 İstanbul Kemerburgaz University, Faculty of Pharmacy, Department of Analytical Chemistry. 2 Yıldız Technical University, Graduate School of Natural & Applied Sciences. 3 Çankırı Karatekin University, Yapraklı Vocational School, Department of Crop & Animal Protection. 4 İstanbul Kemerburgaz University, Faculty of Pharmacy, Department of Biochemistry. 5Near East University, Institute of Environmental Sciences. 6Anadolu University, Faculty of Pharmacy, Department of Pharmacognosy. 7Near East University, Faculty of Pharmacy, Department of Pharmacognosy.
2 Contents A. Stored product protection B. Primary & secondary stored product pests C. Current legislation D. Essential oils as stored product protection agents E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E1. Spotting the active plant species E2. Insecticidal activity screening tests E3. Cholinergic system inhibition E4. Results & Discussion F. Future prospects on EO’s as natural insecticides F1. Methods of Delivery F2. Multiple system inhibitors (Formulation)
3 A. Stored product protection Agricultural produce including grains, legumes, dried fruit, spices, tobacco and medicinal plants has to be stored prior to processing or being distributed to the market. Grains and legumes are usually stored in large amounts and are the most susceptible products to the biotic and abiotic factors. The abiotic factors are mostly related to physical storage conditions such as humidity, temperature, piling, exposure to light and other elements. The biotic factors include fungal, insect, avian and rodent species. Most significant damage to the stored products are caused by fungal and insect species.
4 The primary pests S. granarius, S. oryzae and R. dominica feeds on wheat, corn, legumes and rice. These species can feed on unbroken, whole seeds/beans. The primary pests described here are extremely resistant to very low and high temperatures, humidty and starvation. Primary pests Sitophilus granarius Sitophilus oryzae Rhyzopertha dominica Sitophilus zeamais B. Primary & secondary stored product pests
5 The secondary pests T. castaneum, T. confusum and Oryzaephilus surinamensis feeds on broken wheat, corn, legumes and rice. These species can only feed on broken, whole seeds/beans and flours. Tribolium castaneum Tribolium confusum Oryzaephilus surinamensis Secondary pests B. Primary & secondary stored product pests In the developed countries stored product losses due to insect infestations are estimated to be 9% where as in under developed countries 20% and higher losses are also reported* *Pimentel D. 1991. World resources and food losses to pests. See Ref. 37, pp. 5–11.
6 Post-harvest insects have been controlled through the use of commercial synthetic pesticides. These are either admixed in to the grain as dusts or released in a gaseous state to fumigate stored grain. Most known synthetic pesticides used for protection of stored products are methyl bromide and aluminium phosphide. Alternatively sulphuryl fluoride, carbonyl sulphide, ethane dinitrile and ethyl formate are also used. H Br H H Methyl bromide Methyl bromide was used as a general- purpose fumigant to kill a variety of pests including rats and insects B. Primary & secondary stored product pests
7 Methyl bromide is an extremely toxic gas which have very high vapor pressure and can penetrate through stored product easily. It was used as the main grain fumigant however it has been phased out through the Montreal Protocol on Substances that Deplete the Ozone Layer. The only fumigant now routinely used for protecting grain is aluminium phosphide. Aluminium phosphide is also used to kill rodents and moles and, thus, is highly toxic to vertebrates and can be dangerous for users. Under certain conditions, the phosphine gas generated is also highly explosive. AlP + 3H2O → Al(OH)3 + PH3 Aliminium phosphide Phosphine B. Primary & secondary stored product pests
8 B. Primary & secondary stored product pests Temp. °C Compound Vapor pressure mmHg Hazard Pesticidal use 20 Methyl bromide 1250 Toxic to vertebrates, Toxic to environment, depletes ozone Rodents, Insects 20 Phosphine 26162 Toxic to vertebrates, Explosive, Toxic to environment Rodents, Insects, Fungus 21 Sulfuryl floride 12008 Toxic to vertebrates, greenhouse gas Rodents, Insects 21 Carbonyl sulfide 9034 Toxic to vertebrates, Highly flammable Rodents, Insects -21 Ethanedinitrile 760 Toxic to vertebrates, Toxic to environment, Highly flammable Rodents, Insects, Fungus 20 Ethylformate* 200 Highly flammable Insects
9 B. Primary & secondary stored product pests
10 C. Current legislation Montreal protocol restricts uses of substances that deplete the ozone layer. Most of the countries that signed this protocol phased out/highly restricted uses of pesticides such as methyl bromide. Other local regulations such as the European Pesticide Regulation (EC) No. 1107/2009 supports use of pesticides that are considered less harmful. New regulations like these are expected to provide a special attention to the bio-pesticides that could be considered relatively safe alternative to the synthetic ones !!!
11 C. Current legislation Currently only aluminum phosphide is being used in large scale storages of grains, legumes and corn due to its ease in use. Other alternatives such as ethyl formate, sulfuryl floride and carbonyl sulfide requires skilled use and appropriate application conditions since these are gaseous pest management agents ! The efficacy of these alternatives are sometimes found as questionable by users ! Additionally another problem is that the stored product pests also started to develop resistance against these synthetic pesticides due to their improper uses* *Rajendran, S., Sriranjini V. 2008. “Plant products as fumigants for stored-product insect control.” Journal of Stored Product Research, 44, 126-135. *Shaaya, E., Kostjukovski, M., Eilberg, J., Sukprakarn C. 1997. “Plant oils as Fumigants and Contact Insecticides for the Control of Stored-product Insects.” Journal of Stored Product Research, 33, 1, 7-15.
12 C. Current legislation The problem ! Finding natural alternatives to synthetic fumigant pesticides ! that are : - safe for users - safe for environment - safe for non-target organisms - multipurpose (against fungus, insects, rodents) - capable of peneterating evenly to the stored product - harmless to the stored product (food safety, taste, color, germination properties) - easy to use (have a suitable delivery method) - economic - easy to standardize as a product
13 D. Essential oils as stored product protection agents Inorder to give an idea on the studies that have been conducted until now only essential oils of 57, 138 and 112 taxa were screened only for S. granarius, S. oryzae and S. zeamais ! So there is too much to do in this area ! Most active species are from the following families: Asteraceae (Artemisia, Achillea, Tanacetum), Brassicaceae, Chenopodiastrum (Chenopodium), Ericaceae, Lamiaceae (Origanum, Mentha, Micromeria, Thymus, Salvia), Rutaceae (Citrus, Ruta) , Myrtaceae (Eucalyptus), Zingiberaceae (Curcuma, Zingiber). * K. Polatoğlu, Ö. C. Karakoç, in Essential Oils in Food Preservation - Biologically active essential oils against stored product pests, Academic Press, Elsevier. London, 2016, pp. 39-58
14 D. Essential oils as stored product protection agents * K. Polatoğlu, Ö. C. Karakoç, in Essential Oils in Food Preservation - Biologically active essential oils against stored product pests, Academic Press, Elsevier. London, 2016, pp. 39-58 The major compounds found in the most active oils are : O O Ethyl isovalerate OH Terpinen-4-ol OH Linalool O 1,8-Cineole α-Pinene OH Thymol OH Carvacrol p-Cymene
15 D. Essential oils as stored product protection agents * K. Polatoğlu, Ö. C. Karakoç, in Essential Oils in Food Preservation - Biologically active essential oils against stored product pests, Academic Press, Elsevier. London, 2016, pp. 39-58 The major compounds found in the most active oils are : OH Farnesol H β-SelineneGermacrene D H H Caryophyllene OH O Eugenol O Estragole O O OH Cinnamyl alcohol Safrole
E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E1. Spotting the active plant species In order to work with GRAS plant material we evaluated edibility/medicinal use score of the plant from the literature and local observations The edible/medicinal plants were scored in terms of potential insecticidal activity from the ethnobotanical information and previous insecticidal activity reports on the plants genus The edible/medicinal plants were scored regarding their ecological functions in terms of abundancy in the field, hosting predator insect species, interruption by herbivore insects. The edible/medicinal plants were scored in terms of potential natural products they may contain with insecticidal activity in regards to the previous studies on similar species in the same genus. 16
Natural product score of the plant species E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E1. Spotting the active plant species Plant species to work with ! Ecological score of the plant species Insecticidal activity score of the plant species Edibility/Medicinal score of the plant species (GRAS) 17
E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E2. Insecticidal activity screening tests Two methods of insecticidal activity tests were done on the essential oils that tests effective EO concentration and effective EO exposure time Fumigant insecticidal activity: The EO/EO solution impregnated on filter paper and attached to the tap of an air-tight vial that have a known number of insects and the grain. Insects monitored at 12 h intervals for mortality. Contact insecticidal activity: The EO/EO solution is applied to dorsal surface of each insect by a micro-applicator and a insects are introduced to an air-tight vial that have the grain. Insects are monitored at 12 h intervals for mortality. 18
E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E2. Insecticidal screening tests 19
E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E3. Cholinergic system inhibition 20
E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E3. Cholinergic system inhibition In addition to the insecticidal activity testing; in order to have an idea on the action mechanism of EO’s on the insects, in vitro AChE and BChE inhibition properties of the EO’s were determined according to Ellman’s method*. However in this method acetylcholinesterase obtained from the electric eel and butyrl cholinesterase obtained from equine serum were used. Therefore the obtained results only present a clue whether the insecticidal activity is caused by inhibition of these enzymes. 21 *Ellman, G.L., Courtney, K.D., JR Andres, V., Featherstone, R.M. 1961. A New and rapid colorimetric determination of acetylcholinesterase activity. Biochem. Pharmacol. 7, 88-95.
E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 22 Chrysanthemum coronarium L. Local names: mandilya, krizantem, garland, chop suey Edible/Medicinal Rating: Edible, Medicinal uses reported Insecticidal Activity Rating: Ext. have high activity against Spodoptera littoralis * Ecological Rating: Very abundant in Cyprus and lacks of herbivore attack Natural Product Rating: pyrethrins are obtained from other Chrysanthemum sp. *Shonouda, M.L., Osman, S., Salama, O., Ayoub, A. 2008. Insecticidal effect of Chrysanthemum coronarium L. Flowers on the pest Spodoptera littoralis Boisd and its parasitoid Microplitis rufiventris Kok. with identifying the chemical composition. Journal of Applied Science. 8, 1859-1866.
Samples Değirmenlik (Aerial Parts) Salamis (Aerial Parts) Nicosia/Lefkoşe (Flowers) Main Components in the Essential Oil Camphor 25.2 ± 0.1% Santolinatriene 14.0 ± 0.1% Yomogi alcohol 9.4 ± 0.1% Santolinatriene 21.7 ± 0.1% Camphor 16.4 ± 0.1% Yomogi alcohol 12.6 ± 0.1% cis-chrysanthenyl acetate 10.7 ± 0.1% Bornyl acetate 11.0 ± 0.5% cis-chrysanthenyl acetate 9.9 ± 0.4% Hexadecanoic acid 9.3 ± 0.6% Camphor 9.1 ± 0.3% Phytotoxic Activity (Lemna minor - 10 mg/mL Conc. – Growth Inhibition) 31 % 100.00 % 33.83 % Fumigant Insecticidal Activity (Sitophilus granarius – 10 μL application of 10% (v/v) in acetone – Percent Mortality) 87.82 ± 0.09 % 87.94 ± 0.32 % 97.82 ± 1.79 % E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 23 Chrysanthemum coronarium S N DThe EO’s produced high activity against the insects however they also have phytotoxic activity. This could be problematic if the EO’s also inhibit seed germination !
E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 24 Salvia veneris Hedge Local names: Adaçayı Edible/Medicinal Rating: Edible, Medicinal uses reported Insecticidal Activity Rating: EO’s are reported be active against many insect sp. Ecological Rating: Endemic, Limited to a region and lacks of herbivore attack Natural Product Rating: EO’s contains active monoterpenes/sesquiterpenes* *Perry, N. S., Houghton, P. J., Theobald, A., Jenner, P., Perry, E. K. 2000. In‐vitro Inhibition of Human Erythrocyte Acetylcholinesterase by Salvia lavandulaefolia Essential Oil and Constituent Terpenes. Journal of Pharmacy and Pharmacology. 52, 7, 895-902.
Contact toxicity % Fumigant toxicity % Insect species 24 h 48 h 72 h 72 h Cont. 48 h 48 h Cont. Sitophilus granarius 66.74±0.21 70.00±0.00 70.00±0.00 0.00±0.00 97.63±7.00 0.00±0.00 Sitophilus oryzae 13.01±0.39 53.35±0.19 71.45±2.50 0.00±0.00 100.00±0.00 0.00±0.00 Tribolium castaneum 32.78±1.52 32.78±1.52 32.78±1.52 4.53±1.97 0.00±0.00 0.00±0.00 Tribolium confusum 50.15±1.79 53.50±1.40 53.50±1.40 4.53±1.97 0.00±0.00 0.00±0.00 Rhyzopertha dominica 46.65±0.19 56.84±0.81 67.09±0.94 4.53±1.97 0.00±0.00 0.00±0.00 Oryzaephilus surinamensis 68.84±6.89 77.84±1.91 81.64±2.50 4.53±1.97 90.75±7.00 0.00±0.00 E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 25 Salvia veneris Hedge S. veneris EO was characterized by 1,8 cineole 30.4%, camphor 18.4%, camphene 12.9%, α-pinene 8.2%, borneol 5.8% and β-pinene 5.0 %. AChE Inh. = 85.9% BChE Inh. = 12.2%
E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 26 Crithmum maritimum L. Local names: Girdama Edible/Medicinal Rating: Edible, Medicinal uses reported Insecticidal Activity Rating: EO’s are reported be active against Pheidole pallidula* Ecological Rating: Limited to coastal regions and lacks of herbivore attack Natural Product Rating: EO’s contains active monoterpenes* *Tsoukatou, M., Tsitsimpikou, C., Vagias, C., Roussis, V. 2001. Chemical intra-Mediterranean variation and insecticidal activity of Crithmum maritimum. Z. Naturforsch C. 56, 3-4, 211-215. * Senatore, F., De Feo, V. 1994. Essential oil of a possible new chemotype of Crithmum maritimum L. growing in Campania (Southern Italy). Flav. Fragr. J. 9, 6, 305-307.
Contact toxicity % Fumigant toxicity % Insect species 24 h 48 h 72 h 72 h Cont. 48 h 48 h Cont. Sitophilus granarius 10.00±0.00 23.18±0.26 50.00±0.58 0.00±0.00 100.00±0.00 0.00±0.00 Sitophilus oryzae 46.65±0.19 83.64±0.35 93.30±3.20 0.00±0.00 100.00±0.00 0.00±0.00 Tribolium castaneum 19.31±0.96 19.31±0.96 22.16±1.91 4.53±1.97 0.00±0.00 0.00±0.00 Tribolium confusum 26.20±0.94 33.26±0.21 33.26±0.21 4.53±1.97 0.00±0.00 0.00±0.00 Rhyzopertha dominica 67.09±0.94 67.09±0.94 83.26±7.50 4.53±1.97 4.53±3.41 0.00±0.00 Oryzaephilus surinamensis 56.84±0.81 56.84±0.81 70.33±0.70 4.53±1.97 90.75±7.00 0.00±0.00 E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 27 Crithmum maritimum L. The major components of the oil were identified as γ-terpinene (39.3%), β- phellandrene (22.6%), carvacrol methylether (10.5%) and (Z)-β-ocimene (8.2%). AChE Inh. = 50.3% BChE Inh. = 59.8%
E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 28 Ferulago cypria H. Wolff Local names: Not known Edible/Medicinal Rating: Food additive* & Medicinal uses reported Insecticidal Activity Rating: EO’s are reported be active against Culex pipiens* Ecological Rating: Limited mostly to coastal regions and lacks of herbivore attack Natural Product Rating: Other Ferulago EO’s contain active mono & sesquiterpenes* *Evergetis, E., Michaelakis, A., & Haroutounian, S. A. (2012). Essential oils of Umbelliferae (Apiaceae) family taxa as emerging potent agents for mosquito control. Integrated pest management and pest control—current and future tactics. Janeza Trdine, Croatia, 613-638. *Sadeghi, Ehsan, Farahnaz Karami, and Alireza Etminan. "The Effect of Ferulago angulata (Schlecht) Boiss Essential Oil on Stabilization of Sunflower Oil During Accelerated Storage." Journal of Food Processing and Preservation (2016).
Contact toxicity % 24h Fumigant toxicity % Insect species 24 h, 10 μL (20% (v/v) oil/acetone) 24 h, 10 μL (10% (v/v) oil/acetone) 24 h, 10 μL (5% (v/v) oil/acetone) 24 h Cont. 48 h 10 μL (20% (v/v) oil/acetone) 48 h Cont. Sitophilus granarius 73 % 26 % 0 % 0.00±0.00 * 0 % 0.00±0.00 Sitophilus oryzae 93 % 83 % 0 % 0.00±0.00 * 0 % 0.00±0.00 Tribolium castaneum 13 % 3 % 0 % 0.00±0.00 0 % 0.00±0.00 Tribolium confusum 63 % 40 % 3 % 0.00±0.00 0 % 0.00±0.00 Rhyzopertha dominica 60 % 26 % 6 % 0.00±0.00 0 % 0.00±0.00 E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 29 Ferulago cypria H. Wolff The major components of the oil were identified as limonene (23.8%), α- pinene (11.5%), germacrene D (8.0%) and α-pinene (7.1%). AChE Inh. = 61.5% BChE Inh. = 49.8%
E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 30 Ruta chalepensis L. Local names: Fringed Rue Edible/Medicinal Rating: Medicinal uses reported, some species used as condiment Insecticidal Activity Rating: EO’s are reported be active against Tribolium sp.* Ecological Rating: Abundant and lacks of herbivore attack Natural Product Rating: Contains psoralens *Majdoub, O., Dhen, N., Salaheddine Souguir, U. 2014. Chemical Composition of Ruta chalepensis Essential Oils and their Insecticidal Activity against Tribolium castaneum. Tunisian Journal of Plant Protection. 9, 1, 83-90.
E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 31 Ruta chalepensis L. The main components of the oil were 2- undecanone (21.5 %), 2-nonanone (18.3 %), 2-nonyl acetate (13.2 %) and pregeijerene (3.6 %). Contact toxicity LD90 (µL/insect ) Fumigant toxicity% 48 h Insect species 24 h 48 h 72 h 48 h, 20 μL (10% (v/v) oil/acetone 48 h, 10 μL (10% (v/v) oil/acetone 48 h, Cont. Sitophilus granarius 0.107 0.096 0.096 100.00±0.00 100.00±0.00 0.00±0.00 Sitophilus oryzae 0.090 0.071 0.083 95.47±3.41 56.69±0.34 0.00±0.00 Tribolium castaneum 0.276 0.316 0.311 93.30±5.54 80.69±1.66 0.00±0.00 Tribolium confusum 0.340 0.340 0.347 0.00±0.00 0.00±0.00 0.00±0.00 Rhyzopertha dominica 0.039 0.038 0.038 85.47±3.41 53.35±1.35 0.00±0.00 AChE Inh. = 5.3% BChE Inh. = 42.6%
E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 32 Inula viscosa (L.) Aiton [Syn.: Dittrichia viscosa (L.) Greuter] Local names: konyzos Edible/Medicinal Rating: Medicinal uses reported, No edible uses. Insecticidal Activity Rating: Extracts are reported to be active against aphids* Ecological Rating: Very abundant, lacks of herbivore attack, hosts predator insects Natural Product Rating: EO’s contain active sesquiterpenes* *Mamoci, E., Cavoski, I., Andres, M. F., Díaz, C. E., & Gonzalez-Coloma, A. (2012). Chemical characterization of the aphid antifeedant extracts from Dittrichia viscosa and Ferula communis. Biochemical Systematics and Ecology, 43, 101-107. Al-Qudah, M. A., Al-Jaber, H. I., Mayyas, A. S., Abu-Orabi, S. T., & Abu Zarga, M. H. (2010). Chemical compositions of the essential oil from the Jordanian medicinal plant Dittrichia viscosa. Jordan Journal of Chemistry, 5(4), 343-348.
E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 33 Inula viscosa (L.) Aiton The main components of the oil were (E)-nerolidol (26.6%), selina-6-en-4-ol (8.5%), fokienol (6.8%) and caryophyllene oxide (5.1%). Fumigant toxicity % 48 h (0.01 mL/1mL stock solution, oil/acetone) Insect species 48 h, 10 μL (5% (v/v) stock solution/acetone) 48 h, 10 μL (2.5% (v/v) stock, solution/acetone) 48 h, 10 μL (1% (v/v) stock, solution/acetone) 48 h, Cont. Sitophilus granarius 100.00±0.00 95.47±3.41 4.53±3.41 0.00±0.00 Sitophilus oryzae N.A. N.A. N.A. N.A. Tribolium castaneum N.A. N.A. N.A. N.A. Tribolium confusum N.A. N.A. N.A. N.A. Rhyzopertha dominica N.A. N.A. N.A. N.A. AChE Inh. = 17.5% BChE Inh. = N.A.
E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 34 Achillea biebersteinii Afan. Local names: Yılan Pungu Edible/Medicinal Rating: Medicinal uses reported, No edible uses Insecticidal Activity Rating: EO’s are reported be active against Sitophilus sp., Tribolium sp.* Ecological Rating: Abundant and lacks of herbivore attack Natural Product Rating: EO’s contain active monoterpenes & sesquiterpenes* *Calmasur, O., Kordali, S., Kaya, O., Aslan, I., 2006. Toxicity of essential oil vapours obtained from Achillea spp. to Sitophilus granarius (L.) and Tribolium confusum (Jacquelin du Val). J. Plant Dis. Protect. 113, 1, 37-41.
Samples Mt. Dumluca (Aerial Parts) Mt. Ararat (Aerial Parts) Main Components in the Essential Oil 1,8-Cineole 31.1 % Camphor 14.4% α-Thujone 12.9% p-Cymene 4.6% 1,8-Cineole 30.6% Piperitone 28.9% Camphor 11.7% Phytotoxic Activity (Lemna minor - 10 mg/mL Conc. – Growth Inhibition) 28.74 ± 0.24% 29.95 ± 1.30% Fumigant Insecticidal Activity (Sitophilus granarius – 10 μL application of 10% (v/v) in acetone – Percent Mortality) 1.49 ± 1.12% 36.64 ± 0.12% Contact Insecticidal Activity (Sitophilus granarius – 10 μL application of 10% (v/v) in acetone – Percent Mortality) 3.33 ± 0.00% 62.23 ± 0.04% E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 35 Achillea biebersteinii Afan. Only one chemotype of A. biebersteinii produced considerable activity however it has mild phytotoxic activity! D A
E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 36 A. teretifolia Willd. Local names: Boz yavşan, civan perçemi beyazı, sırçanotu, yavşan otu Edible/Medicinal Rating: Medicinal uses reported, No edible uses Insecticidal Activity Rating: Achillea EO’s are reported to have insecticidal activity Ecological Rating: Abundant and lacks of herbivore attack Natural Product Rating: EO’s contain active monoterpenes* *Demirci, F., Demirci, B., Gürbüz, İ., Yeşilada, E., Başer, K.H.C., 2009. Characterization and Biological Activity of Achillea teretifolia Willd. and A. nobilis L. subsp neilreichii (Kerner) Formanek Essential Oils. Turk. J. Biol. 33, 129-136.
Samples Mt. Dumluca (Aerial Parts) Main Components in the Essential Oil 1,8-Cineole 15.9 % Borneol 8.1% Camphor 7.0% T-cadinol 5.9% Phytotoxic Activity (Lemna minor - 10 mg/mL Conc. – Growth Inhibition) 24.85 ± 9.0% Fumigant Insecticidal Activity (Sitophilus granarius – 10 μL application of 10% (v/v) in acetone – Percent Mortality) 4.32 ± 0.20% Contact Insecticidal Activity (Sitophilus granarius – 10 μL application of 10% (v/v) in acetone – Percent Mortality) 43.32 ± 0.11% E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 37 A. teretifolia Willd. A. teretifolia produced mild insecticidal activity and mild phytotoxic activity!
E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 38 Matricaria chamomilla L. var. recutita (L.) GRIERSON Local names: Papatya Edible/Medicinal Rating: Herbal tea is made, Medicinal uses reported Insecticidal Activity Rating: Matricaria EO’s have insecticidal activity against stored product pests* Ecological Rating: Abundant and lacks of herbivore attack Natural Product Rating: EO’s contain active sesquiterpenes* *Al-Jabr, A. M. (2006). Toxicity and repellency of seven plant essential oils to Oryzaephilus surinamensis (Coleoptera: Silvanidae) and Tribolium castaneum (Coleoptera: Tenebrioidae). Scientific Journal of King Faisal University (Basic and Applied Sciences), 7(1), 49-60. *Roby, M. H. H., Sarhan, M. A., Selim, K. A. H., & Khalel, K. I. (2013). Antioxidant and antimicrobial activities of essential oil and extracts of fennel (Foeniculum vulgare L.) and chamomile (Matricaria chamomilla L.). Industrial crops and products, 44, 437-445.
İstanbul Sample (İ) Contact toxicity % (10 μL, (%10 v/v)) Fumigant toxicity % Insect species 24 h 48 h 72 h 24 h Cont. 24 h Deltamethrin 72 h 72 h Cont. Sitophilus granarius 98.85 ± 2.74 98.85 ± 2.74 99.71 ± 1.72 0.29 ± 1.72 98.85 ± 2.74 74.88 ± 1.35 1.49 ± 1.12 Sitophilus oryzae 98.85 ± 2.74 99.71 ± 1.72 100.00 ± 0.00 0.29 ± 1.72 98.85 ± 2.74 75.57 ± 0.05 0.37 ± 1.12 Tribolium castaneum 14.64 ± 0.60 99.71 ± 1.72 99.71 ± 1.72 0.00 ± 0.00 99.71 ± 1.72 21.09 ± 0.05 0.00 ± 0.00 Tribolium confusum 7.02 ± 1.72 39.79 ± 1.67 68.96 ± 2.07 0.29 ± 1.72 99.71 ± 1.72 90.00 ± 0.00 0.00 ± 0.00 E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species 39 Matricaria chamomilla var. recutita Main component of İstanbul sample are bisabolene oxide (41.9%), Bisabolol oxide A (25.8%) and (E)-β-Farnesene (7.6%). İ E4. Results & Discussion AChE Inh. = 4.7% BChE Inh. = N.A. B
Bursa Sample (B) Contact toxicity % (10 μL, (%20 v/v)) Fumigant toxicity % Insect species 24 h 48 h 72 h 24 h Cont. 24 h Deltamethrin 72 h 72 h Cont. Sitophilus granarius 98.85 ± 2.74 98.85 ± 2.74 98.85 ± 2.74 0.29 ± 1.72 98.85 ± 2.74 96.59 ± 3.45 1.49 ± 1.12 Sitophilus oryzae 99.71 ± 1.72 99.71 ± 1.72 99.71 ± 1.72 0.29 ± 1.72 98.85 ± 2.74 88.12 ± 0,67 0.37 ± 1.12 Tribolium castaneum 0.00 ± 0.00 0.00 ± 0.00 0.29 ± 1.72 0.00 ± 0.00 99.71 ± 1.72 34.39 ± 0.29 0.00 ± 0.00 Tribolium confusum 37.97 ± 1.54 55.10 ± 1.14 56.76 ± 1.10b 0.29 ± 1.72 99.71 ± 1.72 89.11 ± 0.42 0.00 ± 0.00 E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species 40 Matricaria chamomilla var. recutita Main component of İstanbul sample are α-Bisabolol oxide A (47.6%) and (E)-β- Farnesene (12.7%). İ E4. Results & Discussion AChE Inh. = 11.9% BChE Inh. = N.A. B
E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 41 Tanacetum parthenium L. Local names: Papatya Edible/Medicinal Rating: Medicinal uses reported, No edible use Insecticidal Activity Rating: EO’s have insecticidal activity against Spodoptera littoralis* Ecological Rating: Abundant and lacks of herbivore attack Natural Product Rating: EO’s contain active monoterpenes* *Pavela, R., Sajfrtová, M., Sovová, H., Bárnet, M., & Karban, J. (2010). The insecticidal activity of Tanacetum parthenium (L.) Schultz Bip. extracts obtained by supercritical fluid extraction and hydrodistillation. Industrial Crops and Products, 31(3), 449-454.
Contact toxicity % (10 μL, (%20 v/v)) Fumigant toxicity % Insect species 24 h 48 h 72 h 24 h Cont. 24 h Deltamethrin 72 h 72 h Cont. Sitophilus granarius 99.71 ± 1.72 99.71 ± 1.72 99.71 ± 1.72 0.29 ± 1.72 98.85 ± 2.74 100.00 ± 0.00 1.49 ± 1.12 Sitophilus oryzae 98.85 ± 2.74 99.71 ± 1.72 99.71 ± 1.72 0.29 ± 1.72 98.85 ± 2.74 99.63 ± 1.12 0.37 ± 1.12 Tribolium castaneum 1.70 ± 4.25 99.40 ± 0.10 99.71 ± 1.72 0.00 ± 0.00 99.71 ± 1.72 97.63 ± 7.00 0.00 ± 0.00 Tribolium confusum 0.00 ± 0.00 99.40 ± 0.10 99.71 ± 1.72 0.29 ± 1.72 99.71 ± 1.72 96.27 ± 1.84 0.00 ± 0.00 E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species 42 Tanacetum parthenium L. Main component of EO is camphor 36.1% trans-chrysanthenyl acetate 19.5% and camphene 10.7%. İ E4. Results & Discussion AChE Inh. = 20.3% BChE Inh. = N.A.
E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 43 Tanacetum macrophyllum (Waldst. & Kit.) Local names: Not known Edible/Medicinal Rating: Medicinal uses reported, No edible use Insecticidal Activity Rating: EO’s of other species of Tanacetum have insecticidal activity Ecological Rating: Abundant and lacks of herbivore attack Natural Product Rating: EO’s contain active monoterpenes* *Demirci, B., & Başer, K. H. C. (2007). The Essential Oil Composition of Tanacetum macrophyllum (Waldst. et Kit.) Schultz. Bip. Journal of Essential Oil Research, 19(3), 255-257.
E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species 44 Tanacetum macrophyllum (Waldst. & Kit.) E4. Results & Discussion Samples Flower Leaf Main Components in the Essential Oil γ-eudesmol 21.5% (E)-sesquilavandulol 20.3% copaborneol 8.5% copaborneol 14.1% 1,8-cineole 11%, bornyl acetate 9.6% borneol 6.3% Phytotoxic Activity (Lemna minor - 10 mg/mL Conc. – Growth Inhibition) 61.86 ± 5.99% N.A. Contact Insecticidal Activity (Sitophilus granarius – 10 μL application of 10% (v/v) in acetone – Percent Mortality) 50.00 ± 0.00% 88.93 ± 0.09% T. macrophyllum leaf oil have high activity and flower oil have medicore activity against S. granarius however flower oil also have medicore phytotoxic activity.
E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 45 Tanacetum abrotanifolium Druce. Local names: Not known Edible/Medicinal Rating: Medicinal uses reported, No edible use Insecticidal Activity Rating: Extracts have insecticidal activity against stored product pests* Ecological Rating: Abundant in a limited region and lacks of herbivore attack Natural Product Rating: No EO literature prior to study, sesquiterpene lactones *Caliskan, Z. Z., & Goren, N. (2011). Bioassay directed structure elucidation of secondary metabolites in Tanacetum abratonifolium (Compositae). Current Opinion in Biotechnology, 22, S143.
Samples Flower Stem Main Components in the Essential Oil camphor 35.2%, (E)-sesquilavandulol 19.0%, 1,8-cineole 13.5% hexadecanoic acid 41.8%, (E)-sesquilavandulol 16.2%, tetradecanoic acid 6.6% Phytotoxic Activity (Lemna minor - 10 mg/mL Conc. – Growth Inhibition) 38.14 ± 1.20 % N.A Contact Insecticidal Activity (Sitophilus granarius – 10 μL application of 10% (v/v) in acetone – Percent Mortality) 81.30 ± 0.44 % 47.77 ± 0.15 % E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species 46 Tanacetum abrotanifolium Druce. E4. Results & Discussion T. abrotanifolium flower oil have high insecticidal activity against S. granarius however it also have low phytotoxic activity. AChE Inh. = 85.3% (F) BChE Inh. = N.A.
F. Future prospects on EO’s as natural insecticides 47 Another important concern is the application method of the essential oils. Unlike conventional fumigants, essential oils do not produce high vapor pressure and there is a need for a method to provide appropriate vapor pressure for their use as fumigants in large-scale applications. A. Uses of carrier gas such as as nitrogen, oxygen, carbon dioxide, and ethyl formate B. Uses of solid formulations of EO’s with adsorbents (montmorillonite clay, kaolinite clay, diatomaceous earth, silica gel) C. Uses of dust formulations with controlled delivery systems (via encapsulation) D. Uses of new silo designs F1. Methods of Delivery
F. Future prospects on EO’s as natural insecticides 48 F2. Multiple system inhibitors (Formulation) Mixtures of EO’s/EO components that can act on multiple systems should be investigated such as cholinergic, GABA, octapaminergic and mitochondrial systems. Screening of EO’s and EO components for the antagonism of muscarinic, nicotinic, octapaminergic receptors as well as inhibition of enzyme systems such as AChE, BChE, esterases, glutathione S-transferase and monooxygenases is necessary in order to understand action mechanisms.
Conclusions 49 The area of natural pesticides in terms of essential oils offers so many oppurtunities to the scientists working on the essential oils. Essential oils looks very promising in order to develop a new, safe, environmentally friendly fumigation agents. However there are many problems to solve which limits their uses (Stability, Application methodology and limited vapor pressure) This area is still a virgin territory and there are many topics to work on !
50 Teşekkürler… Thank you very much for your attention ! Assist. Prof. Dr. Kaan Polatoglu İstanbul Kemerburgaz University, Faculty of Pharmacy, Department of Analytical Chemistry kaan.polatoglu@kemerburgaz.edu.tr

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ISEO_ORAL_2016_KP_Son_Final

  • 1.
    1 Essential oils aspotential insecticidal agents for protection of stored products Kaan Polatoğlu1, Hüseyin Servi2, Ömer Cem Karakoç3,Yasemin Yücel Yücel4, Salih Gücel5, Betül Demirci6, Kemal Hüsnü Can Başer6,7 1 İstanbul Kemerburgaz University, Faculty of Pharmacy, Department of Analytical Chemistry. 2 Yıldız Technical University, Graduate School of Natural & Applied Sciences. 3 Çankırı Karatekin University, Yapraklı Vocational School, Department of Crop & Animal Protection. 4 İstanbul Kemerburgaz University, Faculty of Pharmacy, Department of Biochemistry. 5Near East University, Institute of Environmental Sciences. 6Anadolu University, Faculty of Pharmacy, Department of Pharmacognosy. 7Near East University, Faculty of Pharmacy, Department of Pharmacognosy.
  • 2.
    2 Contents A. Stored productprotection B. Primary & secondary stored product pests C. Current legislation D. Essential oils as stored product protection agents E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E1. Spotting the active plant species E2. Insecticidal activity screening tests E3. Cholinergic system inhibition E4. Results & Discussion F. Future prospects on EO’s as natural insecticides F1. Methods of Delivery F2. Multiple system inhibitors (Formulation)
  • 3.
    3 A. Stored productprotection Agricultural produce including grains, legumes, dried fruit, spices, tobacco and medicinal plants has to be stored prior to processing or being distributed to the market. Grains and legumes are usually stored in large amounts and are the most susceptible products to the biotic and abiotic factors. The abiotic factors are mostly related to physical storage conditions such as humidity, temperature, piling, exposure to light and other elements. The biotic factors include fungal, insect, avian and rodent species. Most significant damage to the stored products are caused by fungal and insect species.
  • 4.
    4 The primary pestsS. granarius, S. oryzae and R. dominica feeds on wheat, corn, legumes and rice. These species can feed on unbroken, whole seeds/beans. The primary pests described here are extremely resistant to very low and high temperatures, humidty and starvation. Primary pests Sitophilus granarius Sitophilus oryzae Rhyzopertha dominica Sitophilus zeamais B. Primary & secondary stored product pests
  • 5.
    5 The secondary pestsT. castaneum, T. confusum and Oryzaephilus surinamensis feeds on broken wheat, corn, legumes and rice. These species can only feed on broken, whole seeds/beans and flours. Tribolium castaneum Tribolium confusum Oryzaephilus surinamensis Secondary pests B. Primary & secondary stored product pests In the developed countries stored product losses due to insect infestations are estimated to be 9% where as in under developed countries 20% and higher losses are also reported* *Pimentel D. 1991. World resources and food losses to pests. See Ref. 37, pp. 5–11.
  • 6.
    6 Post-harvest insects havebeen controlled through the use of commercial synthetic pesticides. These are either admixed in to the grain as dusts or released in a gaseous state to fumigate stored grain. Most known synthetic pesticides used for protection of stored products are methyl bromide and aluminium phosphide. Alternatively sulphuryl fluoride, carbonyl sulphide, ethane dinitrile and ethyl formate are also used. H Br H H Methyl bromide Methyl bromide was used as a general- purpose fumigant to kill a variety of pests including rats and insects B. Primary & secondary stored product pests
  • 7.
    7 Methyl bromide isan extremely toxic gas which have very high vapor pressure and can penetrate through stored product easily. It was used as the main grain fumigant however it has been phased out through the Montreal Protocol on Substances that Deplete the Ozone Layer. The only fumigant now routinely used for protecting grain is aluminium phosphide. Aluminium phosphide is also used to kill rodents and moles and, thus, is highly toxic to vertebrates and can be dangerous for users. Under certain conditions, the phosphine gas generated is also highly explosive. AlP + 3H2O → Al(OH)3 + PH3 Aliminium phosphide Phosphine B. Primary & secondary stored product pests
  • 8.
    8 B. Primary &secondary stored product pests Temp. °C Compound Vapor pressure mmHg Hazard Pesticidal use 20 Methyl bromide 1250 Toxic to vertebrates, Toxic to environment, depletes ozone Rodents, Insects 20 Phosphine 26162 Toxic to vertebrates, Explosive, Toxic to environment Rodents, Insects, Fungus 21 Sulfuryl floride 12008 Toxic to vertebrates, greenhouse gas Rodents, Insects 21 Carbonyl sulfide 9034 Toxic to vertebrates, Highly flammable Rodents, Insects -21 Ethanedinitrile 760 Toxic to vertebrates, Toxic to environment, Highly flammable Rodents, Insects, Fungus 20 Ethylformate* 200 Highly flammable Insects
  • 9.
    9 B. Primary &secondary stored product pests
  • 10.
    10 C. Current legislation Montrealprotocol restricts uses of substances that deplete the ozone layer. Most of the countries that signed this protocol phased out/highly restricted uses of pesticides such as methyl bromide. Other local regulations such as the European Pesticide Regulation (EC) No. 1107/2009 supports use of pesticides that are considered less harmful. New regulations like these are expected to provide a special attention to the bio-pesticides that could be considered relatively safe alternative to the synthetic ones !!!
  • 11.
    11 C. Current legislation Currentlyonly aluminum phosphide is being used in large scale storages of grains, legumes and corn due to its ease in use. Other alternatives such as ethyl formate, sulfuryl floride and carbonyl sulfide requires skilled use and appropriate application conditions since these are gaseous pest management agents ! The efficacy of these alternatives are sometimes found as questionable by users ! Additionally another problem is that the stored product pests also started to develop resistance against these synthetic pesticides due to their improper uses* *Rajendran, S., Sriranjini V. 2008. “Plant products as fumigants for stored-product insect control.” Journal of Stored Product Research, 44, 126-135. *Shaaya, E., Kostjukovski, M., Eilberg, J., Sukprakarn C. 1997. “Plant oils as Fumigants and Contact Insecticides for the Control of Stored-product Insects.” Journal of Stored Product Research, 33, 1, 7-15.
  • 12.
    12 C. Current legislation Theproblem ! Finding natural alternatives to synthetic fumigant pesticides ! that are : - safe for users - safe for environment - safe for non-target organisms - multipurpose (against fungus, insects, rodents) - capable of peneterating evenly to the stored product - harmless to the stored product (food safety, taste, color, germination properties) - easy to use (have a suitable delivery method) - economic - easy to standardize as a product
  • 13.
    13 D. Essential oilsas stored product protection agents Inorder to give an idea on the studies that have been conducted until now only essential oils of 57, 138 and 112 taxa were screened only for S. granarius, S. oryzae and S. zeamais ! So there is too much to do in this area ! Most active species are from the following families: Asteraceae (Artemisia, Achillea, Tanacetum), Brassicaceae, Chenopodiastrum (Chenopodium), Ericaceae, Lamiaceae (Origanum, Mentha, Micromeria, Thymus, Salvia), Rutaceae (Citrus, Ruta) , Myrtaceae (Eucalyptus), Zingiberaceae (Curcuma, Zingiber). * K. Polatoğlu, Ö. C. Karakoç, in Essential Oils in Food Preservation - Biologically active essential oils against stored product pests, Academic Press, Elsevier. London, 2016, pp. 39-58
  • 14.
    14 D. Essential oilsas stored product protection agents * K. Polatoğlu, Ö. C. Karakoç, in Essential Oils in Food Preservation - Biologically active essential oils against stored product pests, Academic Press, Elsevier. London, 2016, pp. 39-58 The major compounds found in the most active oils are : O O Ethyl isovalerate OH Terpinen-4-ol OH Linalool O 1,8-Cineole α-Pinene OH Thymol OH Carvacrol p-Cymene
  • 15.
    15 D. Essential oilsas stored product protection agents * K. Polatoğlu, Ö. C. Karakoç, in Essential Oils in Food Preservation - Biologically active essential oils against stored product pests, Academic Press, Elsevier. London, 2016, pp. 39-58 The major compounds found in the most active oils are : OH Farnesol H β-SelineneGermacrene D H H Caryophyllene OH O Eugenol O Estragole O O OH Cinnamyl alcohol Safrole
  • 16.
    E. Insecticidal activityscreening studies of EO from Turkish & Cypriot plant species E1. Spotting the active plant species In order to work with GRAS plant material we evaluated edibility/medicinal use score of the plant from the literature and local observations The edible/medicinal plants were scored in terms of potential insecticidal activity from the ethnobotanical information and previous insecticidal activity reports on the plants genus The edible/medicinal plants were scored regarding their ecological functions in terms of abundancy in the field, hosting predator insect species, interruption by herbivore insects. The edible/medicinal plants were scored in terms of potential natural products they may contain with insecticidal activity in regards to the previous studies on similar species in the same genus. 16
  • 17.
    Natural product scoreof the plant species E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E1. Spotting the active plant species Plant species to work with ! Ecological score of the plant species Insecticidal activity score of the plant species Edibility/Medicinal score of the plant species (GRAS) 17
  • 18.
    E. Insecticidal activityscreening studies of EO from Turkish & Cypriot plant species E2. Insecticidal activity screening tests Two methods of insecticidal activity tests were done on the essential oils that tests effective EO concentration and effective EO exposure time Fumigant insecticidal activity: The EO/EO solution impregnated on filter paper and attached to the tap of an air-tight vial that have a known number of insects and the grain. Insects monitored at 12 h intervals for mortality. Contact insecticidal activity: The EO/EO solution is applied to dorsal surface of each insect by a micro-applicator and a insects are introduced to an air-tight vial that have the grain. Insects are monitored at 12 h intervals for mortality. 18
  • 19.
    E. Insecticidal activityscreening studies of EO from Turkish & Cypriot plant species E2. Insecticidal screening tests 19
  • 20.
    E. Insecticidal activityscreening studies of EO from Turkish & Cypriot plant species E3. Cholinergic system inhibition 20
  • 21.
    E. Insecticidal activityscreening studies of EO from Turkish & Cypriot plant species E3. Cholinergic system inhibition In addition to the insecticidal activity testing; in order to have an idea on the action mechanism of EO’s on the insects, in vitro AChE and BChE inhibition properties of the EO’s were determined according to Ellman’s method*. However in this method acetylcholinesterase obtained from the electric eel and butyrl cholinesterase obtained from equine serum were used. Therefore the obtained results only present a clue whether the insecticidal activity is caused by inhibition of these enzymes. 21 *Ellman, G.L., Courtney, K.D., JR Andres, V., Featherstone, R.M. 1961. A New and rapid colorimetric determination of acetylcholinesterase activity. Biochem. Pharmacol. 7, 88-95.
  • 22.
    E. Insecticidal activityscreening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 22 Chrysanthemum coronarium L. Local names: mandilya, krizantem, garland, chop suey Edible/Medicinal Rating: Edible, Medicinal uses reported Insecticidal Activity Rating: Ext. have high activity against Spodoptera littoralis * Ecological Rating: Very abundant in Cyprus and lacks of herbivore attack Natural Product Rating: pyrethrins are obtained from other Chrysanthemum sp. *Shonouda, M.L., Osman, S., Salama, O., Ayoub, A. 2008. Insecticidal effect of Chrysanthemum coronarium L. Flowers on the pest Spodoptera littoralis Boisd and its parasitoid Microplitis rufiventris Kok. with identifying the chemical composition. Journal of Applied Science. 8, 1859-1866.
  • 23.
    Samples Değirmenlik (Aerial Parts) Salamis(Aerial Parts) Nicosia/Lefkoşe (Flowers) Main Components in the Essential Oil Camphor 25.2 ± 0.1% Santolinatriene 14.0 ± 0.1% Yomogi alcohol 9.4 ± 0.1% Santolinatriene 21.7 ± 0.1% Camphor 16.4 ± 0.1% Yomogi alcohol 12.6 ± 0.1% cis-chrysanthenyl acetate 10.7 ± 0.1% Bornyl acetate 11.0 ± 0.5% cis-chrysanthenyl acetate 9.9 ± 0.4% Hexadecanoic acid 9.3 ± 0.6% Camphor 9.1 ± 0.3% Phytotoxic Activity (Lemna minor - 10 mg/mL Conc. – Growth Inhibition) 31 % 100.00 % 33.83 % Fumigant Insecticidal Activity (Sitophilus granarius – 10 μL application of 10% (v/v) in acetone – Percent Mortality) 87.82 ± 0.09 % 87.94 ± 0.32 % 97.82 ± 1.79 % E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 23 Chrysanthemum coronarium S N DThe EO’s produced high activity against the insects however they also have phytotoxic activity. This could be problematic if the EO’s also inhibit seed germination !
  • 24.
    E. Insecticidal activityscreening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 24 Salvia veneris Hedge Local names: Adaçayı Edible/Medicinal Rating: Edible, Medicinal uses reported Insecticidal Activity Rating: EO’s are reported be active against many insect sp. Ecological Rating: Endemic, Limited to a region and lacks of herbivore attack Natural Product Rating: EO’s contains active monoterpenes/sesquiterpenes* *Perry, N. S., Houghton, P. J., Theobald, A., Jenner, P., Perry, E. K. 2000. In‐vitro Inhibition of Human Erythrocyte Acetylcholinesterase by Salvia lavandulaefolia Essential Oil and Constituent Terpenes. Journal of Pharmacy and Pharmacology. 52, 7, 895-902.
  • 25.
    Contact toxicity %Fumigant toxicity % Insect species 24 h 48 h 72 h 72 h Cont. 48 h 48 h Cont. Sitophilus granarius 66.74±0.21 70.00±0.00 70.00±0.00 0.00±0.00 97.63±7.00 0.00±0.00 Sitophilus oryzae 13.01±0.39 53.35±0.19 71.45±2.50 0.00±0.00 100.00±0.00 0.00±0.00 Tribolium castaneum 32.78±1.52 32.78±1.52 32.78±1.52 4.53±1.97 0.00±0.00 0.00±0.00 Tribolium confusum 50.15±1.79 53.50±1.40 53.50±1.40 4.53±1.97 0.00±0.00 0.00±0.00 Rhyzopertha dominica 46.65±0.19 56.84±0.81 67.09±0.94 4.53±1.97 0.00±0.00 0.00±0.00 Oryzaephilus surinamensis 68.84±6.89 77.84±1.91 81.64±2.50 4.53±1.97 90.75±7.00 0.00±0.00 E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 25 Salvia veneris Hedge S. veneris EO was characterized by 1,8 cineole 30.4%, camphor 18.4%, camphene 12.9%, α-pinene 8.2%, borneol 5.8% and β-pinene 5.0 %. AChE Inh. = 85.9% BChE Inh. = 12.2%
  • 26.
    E. Insecticidal activityscreening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 26 Crithmum maritimum L. Local names: Girdama Edible/Medicinal Rating: Edible, Medicinal uses reported Insecticidal Activity Rating: EO’s are reported be active against Pheidole pallidula* Ecological Rating: Limited to coastal regions and lacks of herbivore attack Natural Product Rating: EO’s contains active monoterpenes* *Tsoukatou, M., Tsitsimpikou, C., Vagias, C., Roussis, V. 2001. Chemical intra-Mediterranean variation and insecticidal activity of Crithmum maritimum. Z. Naturforsch C. 56, 3-4, 211-215. * Senatore, F., De Feo, V. 1994. Essential oil of a possible new chemotype of Crithmum maritimum L. growing in Campania (Southern Italy). Flav. Fragr. J. 9, 6, 305-307.
  • 27.
    Contact toxicity %Fumigant toxicity % Insect species 24 h 48 h 72 h 72 h Cont. 48 h 48 h Cont. Sitophilus granarius 10.00±0.00 23.18±0.26 50.00±0.58 0.00±0.00 100.00±0.00 0.00±0.00 Sitophilus oryzae 46.65±0.19 83.64±0.35 93.30±3.20 0.00±0.00 100.00±0.00 0.00±0.00 Tribolium castaneum 19.31±0.96 19.31±0.96 22.16±1.91 4.53±1.97 0.00±0.00 0.00±0.00 Tribolium confusum 26.20±0.94 33.26±0.21 33.26±0.21 4.53±1.97 0.00±0.00 0.00±0.00 Rhyzopertha dominica 67.09±0.94 67.09±0.94 83.26±7.50 4.53±1.97 4.53±3.41 0.00±0.00 Oryzaephilus surinamensis 56.84±0.81 56.84±0.81 70.33±0.70 4.53±1.97 90.75±7.00 0.00±0.00 E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 27 Crithmum maritimum L. The major components of the oil were identified as γ-terpinene (39.3%), β- phellandrene (22.6%), carvacrol methylether (10.5%) and (Z)-β-ocimene (8.2%). AChE Inh. = 50.3% BChE Inh. = 59.8%
  • 28.
    E. Insecticidal activityscreening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 28 Ferulago cypria H. Wolff Local names: Not known Edible/Medicinal Rating: Food additive* & Medicinal uses reported Insecticidal Activity Rating: EO’s are reported be active against Culex pipiens* Ecological Rating: Limited mostly to coastal regions and lacks of herbivore attack Natural Product Rating: Other Ferulago EO’s contain active mono & sesquiterpenes* *Evergetis, E., Michaelakis, A., & Haroutounian, S. A. (2012). Essential oils of Umbelliferae (Apiaceae) family taxa as emerging potent agents for mosquito control. Integrated pest management and pest control—current and future tactics. Janeza Trdine, Croatia, 613-638. *Sadeghi, Ehsan, Farahnaz Karami, and Alireza Etminan. "The Effect of Ferulago angulata (Schlecht) Boiss Essential Oil on Stabilization of Sunflower Oil During Accelerated Storage." Journal of Food Processing and Preservation (2016).
  • 29.
    Contact toxicity %24h Fumigant toxicity % Insect species 24 h, 10 μL (20% (v/v) oil/acetone) 24 h, 10 μL (10% (v/v) oil/acetone) 24 h, 10 μL (5% (v/v) oil/acetone) 24 h Cont. 48 h 10 μL (20% (v/v) oil/acetone) 48 h Cont. Sitophilus granarius 73 % 26 % 0 % 0.00±0.00 * 0 % 0.00±0.00 Sitophilus oryzae 93 % 83 % 0 % 0.00±0.00 * 0 % 0.00±0.00 Tribolium castaneum 13 % 3 % 0 % 0.00±0.00 0 % 0.00±0.00 Tribolium confusum 63 % 40 % 3 % 0.00±0.00 0 % 0.00±0.00 Rhyzopertha dominica 60 % 26 % 6 % 0.00±0.00 0 % 0.00±0.00 E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 29 Ferulago cypria H. Wolff The major components of the oil were identified as limonene (23.8%), α- pinene (11.5%), germacrene D (8.0%) and α-pinene (7.1%). AChE Inh. = 61.5% BChE Inh. = 49.8%
  • 30.
    E. Insecticidal activityscreening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 30 Ruta chalepensis L. Local names: Fringed Rue Edible/Medicinal Rating: Medicinal uses reported, some species used as condiment Insecticidal Activity Rating: EO’s are reported be active against Tribolium sp.* Ecological Rating: Abundant and lacks of herbivore attack Natural Product Rating: Contains psoralens *Majdoub, O., Dhen, N., Salaheddine Souguir, U. 2014. Chemical Composition of Ruta chalepensis Essential Oils and their Insecticidal Activity against Tribolium castaneum. Tunisian Journal of Plant Protection. 9, 1, 83-90.
  • 31.
    E. Insecticidal activityscreening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 31 Ruta chalepensis L. The main components of the oil were 2- undecanone (21.5 %), 2-nonanone (18.3 %), 2-nonyl acetate (13.2 %) and pregeijerene (3.6 %). Contact toxicity LD90 (µL/insect ) Fumigant toxicity% 48 h Insect species 24 h 48 h 72 h 48 h, 20 μL (10% (v/v) oil/acetone 48 h, 10 μL (10% (v/v) oil/acetone 48 h, Cont. Sitophilus granarius 0.107 0.096 0.096 100.00±0.00 100.00±0.00 0.00±0.00 Sitophilus oryzae 0.090 0.071 0.083 95.47±3.41 56.69±0.34 0.00±0.00 Tribolium castaneum 0.276 0.316 0.311 93.30±5.54 80.69±1.66 0.00±0.00 Tribolium confusum 0.340 0.340 0.347 0.00±0.00 0.00±0.00 0.00±0.00 Rhyzopertha dominica 0.039 0.038 0.038 85.47±3.41 53.35±1.35 0.00±0.00 AChE Inh. = 5.3% BChE Inh. = 42.6%
  • 32.
    E. Insecticidal activityscreening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 32 Inula viscosa (L.) Aiton [Syn.: Dittrichia viscosa (L.) Greuter] Local names: konyzos Edible/Medicinal Rating: Medicinal uses reported, No edible uses. Insecticidal Activity Rating: Extracts are reported to be active against aphids* Ecological Rating: Very abundant, lacks of herbivore attack, hosts predator insects Natural Product Rating: EO’s contain active sesquiterpenes* *Mamoci, E., Cavoski, I., Andres, M. F., Díaz, C. E., & Gonzalez-Coloma, A. (2012). Chemical characterization of the aphid antifeedant extracts from Dittrichia viscosa and Ferula communis. Biochemical Systematics and Ecology, 43, 101-107. Al-Qudah, M. A., Al-Jaber, H. I., Mayyas, A. S., Abu-Orabi, S. T., & Abu Zarga, M. H. (2010). Chemical compositions of the essential oil from the Jordanian medicinal plant Dittrichia viscosa. Jordan Journal of Chemistry, 5(4), 343-348.
  • 33.
    E. Insecticidal activityscreening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 33 Inula viscosa (L.) Aiton The main components of the oil were (E)-nerolidol (26.6%), selina-6-en-4-ol (8.5%), fokienol (6.8%) and caryophyllene oxide (5.1%). Fumigant toxicity % 48 h (0.01 mL/1mL stock solution, oil/acetone) Insect species 48 h, 10 μL (5% (v/v) stock solution/acetone) 48 h, 10 μL (2.5% (v/v) stock, solution/acetone) 48 h, 10 μL (1% (v/v) stock, solution/acetone) 48 h, Cont. Sitophilus granarius 100.00±0.00 95.47±3.41 4.53±3.41 0.00±0.00 Sitophilus oryzae N.A. N.A. N.A. N.A. Tribolium castaneum N.A. N.A. N.A. N.A. Tribolium confusum N.A. N.A. N.A. N.A. Rhyzopertha dominica N.A. N.A. N.A. N.A. AChE Inh. = 17.5% BChE Inh. = N.A.
  • 34.
    E. Insecticidal activityscreening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 34 Achillea biebersteinii Afan. Local names: Yılan Pungu Edible/Medicinal Rating: Medicinal uses reported, No edible uses Insecticidal Activity Rating: EO’s are reported be active against Sitophilus sp., Tribolium sp.* Ecological Rating: Abundant and lacks of herbivore attack Natural Product Rating: EO’s contain active monoterpenes & sesquiterpenes* *Calmasur, O., Kordali, S., Kaya, O., Aslan, I., 2006. Toxicity of essential oil vapours obtained from Achillea spp. to Sitophilus granarius (L.) and Tribolium confusum (Jacquelin du Val). J. Plant Dis. Protect. 113, 1, 37-41.
  • 35.
    Samples Mt. Dumluca(Aerial Parts) Mt. Ararat (Aerial Parts) Main Components in the Essential Oil 1,8-Cineole 31.1 % Camphor 14.4% α-Thujone 12.9% p-Cymene 4.6% 1,8-Cineole 30.6% Piperitone 28.9% Camphor 11.7% Phytotoxic Activity (Lemna minor - 10 mg/mL Conc. – Growth Inhibition) 28.74 ± 0.24% 29.95 ± 1.30% Fumigant Insecticidal Activity (Sitophilus granarius – 10 μL application of 10% (v/v) in acetone – Percent Mortality) 1.49 ± 1.12% 36.64 ± 0.12% Contact Insecticidal Activity (Sitophilus granarius – 10 μL application of 10% (v/v) in acetone – Percent Mortality) 3.33 ± 0.00% 62.23 ± 0.04% E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 35 Achillea biebersteinii Afan. Only one chemotype of A. biebersteinii produced considerable activity however it has mild phytotoxic activity! D A
  • 36.
    E. Insecticidal activityscreening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 36 A. teretifolia Willd. Local names: Boz yavşan, civan perçemi beyazı, sırçanotu, yavşan otu Edible/Medicinal Rating: Medicinal uses reported, No edible uses Insecticidal Activity Rating: Achillea EO’s are reported to have insecticidal activity Ecological Rating: Abundant and lacks of herbivore attack Natural Product Rating: EO’s contain active monoterpenes* *Demirci, F., Demirci, B., Gürbüz, İ., Yeşilada, E., Başer, K.H.C., 2009. Characterization and Biological Activity of Achillea teretifolia Willd. and A. nobilis L. subsp neilreichii (Kerner) Formanek Essential Oils. Turk. J. Biol. 33, 129-136.
  • 37.
    Samples Mt. Dumluca(Aerial Parts) Main Components in the Essential Oil 1,8-Cineole 15.9 % Borneol 8.1% Camphor 7.0% T-cadinol 5.9% Phytotoxic Activity (Lemna minor - 10 mg/mL Conc. – Growth Inhibition) 24.85 ± 9.0% Fumigant Insecticidal Activity (Sitophilus granarius – 10 μL application of 10% (v/v) in acetone – Percent Mortality) 4.32 ± 0.20% Contact Insecticidal Activity (Sitophilus granarius – 10 μL application of 10% (v/v) in acetone – Percent Mortality) 43.32 ± 0.11% E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 37 A. teretifolia Willd. A. teretifolia produced mild insecticidal activity and mild phytotoxic activity!
  • 38.
    E. Insecticidal activityscreening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 38 Matricaria chamomilla L. var. recutita (L.) GRIERSON Local names: Papatya Edible/Medicinal Rating: Herbal tea is made, Medicinal uses reported Insecticidal Activity Rating: Matricaria EO’s have insecticidal activity against stored product pests* Ecological Rating: Abundant and lacks of herbivore attack Natural Product Rating: EO’s contain active sesquiterpenes* *Al-Jabr, A. M. (2006). Toxicity and repellency of seven plant essential oils to Oryzaephilus surinamensis (Coleoptera: Silvanidae) and Tribolium castaneum (Coleoptera: Tenebrioidae). Scientific Journal of King Faisal University (Basic and Applied Sciences), 7(1), 49-60. *Roby, M. H. H., Sarhan, M. A., Selim, K. A. H., & Khalel, K. I. (2013). Antioxidant and antimicrobial activities of essential oil and extracts of fennel (Foeniculum vulgare L.) and chamomile (Matricaria chamomilla L.). Industrial crops and products, 44, 437-445.
  • 39.
    İstanbul Sample (İ)Contact toxicity % (10 μL, (%10 v/v)) Fumigant toxicity % Insect species 24 h 48 h 72 h 24 h Cont. 24 h Deltamethrin 72 h 72 h Cont. Sitophilus granarius 98.85 ± 2.74 98.85 ± 2.74 99.71 ± 1.72 0.29 ± 1.72 98.85 ± 2.74 74.88 ± 1.35 1.49 ± 1.12 Sitophilus oryzae 98.85 ± 2.74 99.71 ± 1.72 100.00 ± 0.00 0.29 ± 1.72 98.85 ± 2.74 75.57 ± 0.05 0.37 ± 1.12 Tribolium castaneum 14.64 ± 0.60 99.71 ± 1.72 99.71 ± 1.72 0.00 ± 0.00 99.71 ± 1.72 21.09 ± 0.05 0.00 ± 0.00 Tribolium confusum 7.02 ± 1.72 39.79 ± 1.67 68.96 ± 2.07 0.29 ± 1.72 99.71 ± 1.72 90.00 ± 0.00 0.00 ± 0.00 E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species 39 Matricaria chamomilla var. recutita Main component of İstanbul sample are bisabolene oxide (41.9%), Bisabolol oxide A (25.8%) and (E)-β-Farnesene (7.6%). İ E4. Results & Discussion AChE Inh. = 4.7% BChE Inh. = N.A. B
  • 40.
    Bursa Sample (B)Contact toxicity % (10 μL, (%20 v/v)) Fumigant toxicity % Insect species 24 h 48 h 72 h 24 h Cont. 24 h Deltamethrin 72 h 72 h Cont. Sitophilus granarius 98.85 ± 2.74 98.85 ± 2.74 98.85 ± 2.74 0.29 ± 1.72 98.85 ± 2.74 96.59 ± 3.45 1.49 ± 1.12 Sitophilus oryzae 99.71 ± 1.72 99.71 ± 1.72 99.71 ± 1.72 0.29 ± 1.72 98.85 ± 2.74 88.12 ± 0,67 0.37 ± 1.12 Tribolium castaneum 0.00 ± 0.00 0.00 ± 0.00 0.29 ± 1.72 0.00 ± 0.00 99.71 ± 1.72 34.39 ± 0.29 0.00 ± 0.00 Tribolium confusum 37.97 ± 1.54 55.10 ± 1.14 56.76 ± 1.10b 0.29 ± 1.72 99.71 ± 1.72 89.11 ± 0.42 0.00 ± 0.00 E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species 40 Matricaria chamomilla var. recutita Main component of İstanbul sample are α-Bisabolol oxide A (47.6%) and (E)-β- Farnesene (12.7%). İ E4. Results & Discussion AChE Inh. = 11.9% BChE Inh. = N.A. B
  • 41.
    E. Insecticidal activityscreening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 41 Tanacetum parthenium L. Local names: Papatya Edible/Medicinal Rating: Medicinal uses reported, No edible use Insecticidal Activity Rating: EO’s have insecticidal activity against Spodoptera littoralis* Ecological Rating: Abundant and lacks of herbivore attack Natural Product Rating: EO’s contain active monoterpenes* *Pavela, R., Sajfrtová, M., Sovová, H., Bárnet, M., & Karban, J. (2010). The insecticidal activity of Tanacetum parthenium (L.) Schultz Bip. extracts obtained by supercritical fluid extraction and hydrodistillation. Industrial Crops and Products, 31(3), 449-454.
  • 42.
    Contact toxicity %(10 μL, (%20 v/v)) Fumigant toxicity % Insect species 24 h 48 h 72 h 24 h Cont. 24 h Deltamethrin 72 h 72 h Cont. Sitophilus granarius 99.71 ± 1.72 99.71 ± 1.72 99.71 ± 1.72 0.29 ± 1.72 98.85 ± 2.74 100.00 ± 0.00 1.49 ± 1.12 Sitophilus oryzae 98.85 ± 2.74 99.71 ± 1.72 99.71 ± 1.72 0.29 ± 1.72 98.85 ± 2.74 99.63 ± 1.12 0.37 ± 1.12 Tribolium castaneum 1.70 ± 4.25 99.40 ± 0.10 99.71 ± 1.72 0.00 ± 0.00 99.71 ± 1.72 97.63 ± 7.00 0.00 ± 0.00 Tribolium confusum 0.00 ± 0.00 99.40 ± 0.10 99.71 ± 1.72 0.29 ± 1.72 99.71 ± 1.72 96.27 ± 1.84 0.00 ± 0.00 E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species 42 Tanacetum parthenium L. Main component of EO is camphor 36.1% trans-chrysanthenyl acetate 19.5% and camphene 10.7%. İ E4. Results & Discussion AChE Inh. = 20.3% BChE Inh. = N.A.
  • 43.
    E. Insecticidal activityscreening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 43 Tanacetum macrophyllum (Waldst. & Kit.) Local names: Not known Edible/Medicinal Rating: Medicinal uses reported, No edible use Insecticidal Activity Rating: EO’s of other species of Tanacetum have insecticidal activity Ecological Rating: Abundant and lacks of herbivore attack Natural Product Rating: EO’s contain active monoterpenes* *Demirci, B., & Başer, K. H. C. (2007). The Essential Oil Composition of Tanacetum macrophyllum (Waldst. et Kit.) Schultz. Bip. Journal of Essential Oil Research, 19(3), 255-257.
  • 44.
    E. Insecticidal activityscreening studies of EO from Turkish & Cypriot plant species 44 Tanacetum macrophyllum (Waldst. & Kit.) E4. Results & Discussion Samples Flower Leaf Main Components in the Essential Oil γ-eudesmol 21.5% (E)-sesquilavandulol 20.3% copaborneol 8.5% copaborneol 14.1% 1,8-cineole 11%, bornyl acetate 9.6% borneol 6.3% Phytotoxic Activity (Lemna minor - 10 mg/mL Conc. – Growth Inhibition) 61.86 ± 5.99% N.A. Contact Insecticidal Activity (Sitophilus granarius – 10 μL application of 10% (v/v) in acetone – Percent Mortality) 50.00 ± 0.00% 88.93 ± 0.09% T. macrophyllum leaf oil have high activity and flower oil have medicore activity against S. granarius however flower oil also have medicore phytotoxic activity.
  • 45.
    E. Insecticidal activityscreening studies of EO from Turkish & Cypriot plant species E4. Results & Discussion 45 Tanacetum abrotanifolium Druce. Local names: Not known Edible/Medicinal Rating: Medicinal uses reported, No edible use Insecticidal Activity Rating: Extracts have insecticidal activity against stored product pests* Ecological Rating: Abundant in a limited region and lacks of herbivore attack Natural Product Rating: No EO literature prior to study, sesquiterpene lactones *Caliskan, Z. Z., & Goren, N. (2011). Bioassay directed structure elucidation of secondary metabolites in Tanacetum abratonifolium (Compositae). Current Opinion in Biotechnology, 22, S143.
  • 46.
    Samples Flower Stem MainComponents in the Essential Oil camphor 35.2%, (E)-sesquilavandulol 19.0%, 1,8-cineole 13.5% hexadecanoic acid 41.8%, (E)-sesquilavandulol 16.2%, tetradecanoic acid 6.6% Phytotoxic Activity (Lemna minor - 10 mg/mL Conc. – Growth Inhibition) 38.14 ± 1.20 % N.A Contact Insecticidal Activity (Sitophilus granarius – 10 μL application of 10% (v/v) in acetone – Percent Mortality) 81.30 ± 0.44 % 47.77 ± 0.15 % E. Insecticidal activity screening studies of EO from Turkish & Cypriot plant species 46 Tanacetum abrotanifolium Druce. E4. Results & Discussion T. abrotanifolium flower oil have high insecticidal activity against S. granarius however it also have low phytotoxic activity. AChE Inh. = 85.3% (F) BChE Inh. = N.A.
  • 47.
    F. Future prospectson EO’s as natural insecticides 47 Another important concern is the application method of the essential oils. Unlike conventional fumigants, essential oils do not produce high vapor pressure and there is a need for a method to provide appropriate vapor pressure for their use as fumigants in large-scale applications. A. Uses of carrier gas such as as nitrogen, oxygen, carbon dioxide, and ethyl formate B. Uses of solid formulations of EO’s with adsorbents (montmorillonite clay, kaolinite clay, diatomaceous earth, silica gel) C. Uses of dust formulations with controlled delivery systems (via encapsulation) D. Uses of new silo designs F1. Methods of Delivery
  • 48.
    F. Future prospectson EO’s as natural insecticides 48 F2. Multiple system inhibitors (Formulation) Mixtures of EO’s/EO components that can act on multiple systems should be investigated such as cholinergic, GABA, octapaminergic and mitochondrial systems. Screening of EO’s and EO components for the antagonism of muscarinic, nicotinic, octapaminergic receptors as well as inhibition of enzyme systems such as AChE, BChE, esterases, glutathione S-transferase and monooxygenases is necessary in order to understand action mechanisms.
  • 49.
    Conclusions 49 The area ofnatural pesticides in terms of essential oils offers so many oppurtunities to the scientists working on the essential oils. Essential oils looks very promising in order to develop a new, safe, environmentally friendly fumigation agents. However there are many problems to solve which limits their uses (Stability, Application methodology and limited vapor pressure) This area is still a virgin territory and there are many topics to work on !
  • 50.
    50 Teşekkürler… Thank you very muchfor your attention ! Assist. Prof. Dr. Kaan Polatoglu İstanbul Kemerburgaz University, Faculty of Pharmacy, Department of Analytical Chemistry kaan.polatoglu@kemerburgaz.edu.tr