Entomotoxic effects of synthesized aluminum oxide nanoparticles against Sitophilus oryzae and their toxicological effects on albino rats.

Recently, nanoparticles are emerging as a potential alternative to synthetic pesticides for protection against stored-product insect pests, such as the rice weevil Sitophilus oryzae; however, the toxic effects of nanoparticles on nontarget organisms are not yet understood. Therefore, we investigated the insecticidal effects of synthesized aluminum oxide nanoparticles (Al2O3-NPs) on S. oryzae, as well as their potential toxicity in albino rats. S. oryzae mortality increased as the period of Al2O3-NP exposure increased; 100% mortality was reached at 8000 mg Al2O3-NPs/kg of wheat grain after 7 days of exposure. After 60 days of exposure, all tested Al2O3-NPs concentrations (1000, 2000, 4000, and 8000 mg/kg grain) significantly reduced the number of S. oryzae offspring in a dose-dependent manner. In albino rats, exposure to the LC90 of Al2O3-NPs in a treated diet caused a significant decrease in total body weight and an increase in liver weight in a subacute toxicity test. Moreover, Al2O3-NP treatment elevated the levels of alanine aminotransferase, aspartate aminotransferase, and creatinine in exposed rats relative to control rats, while the uric acid levels of treated rats decreased. Histopathological analysis also revealed various hepatic and renal lesions in treated rats. In summary, although Al2O3-NPs have insecticidal effects, they also have hazardous toxicological effects on rats. Therefore, if Al2O3-NPs are used in the current powder form to protect stored products, they may cause adverse effects to workers and consumers. Further research will be required to develop new nanoformulations with increased safety and potency before these nanoparticles can be used in stored-product pest control.

Clove Buds Essential Oil: The Impact of Grinding on the Chemical Composition and Its Biological Activities Involved in Consumer's Health Security.

This study is aimed at identifying the chemical composition of the essential oil extracted from the Syzygium aromaticum seeds, as well as investigating its biological activities, insecticide effect, and allelopathic properties. The extraction yield was about 14.3 and 7.14% for grounded and ungrounded seeds, respectively. The GC-MS analysis allowed the identification of 17 heterogeneous compounds, including eugenol (68.7-87.4%), as major compound, cyperene (20.5-7.2%), phenethyl isovalerate (6.4-3.6%), and cis-thujopsene (1.9-0.8%), respectively, for grounded and ungrounded seeds. Concerning the antibacterial activity, the diameter of the inhibition zone reached 35 mm when the essential oil extracted from grounded seeds was applied against Escherichia coli. Regarding the antioxidant activity via the DPPH radical scavenging test, the IC50 varied from 1.2 ± 0.1 to 2.8 ± 0.5 µg/mL. With respect to reducing power, the efficient concentration EC50 ranged from 32 to 50 µg/mL. The essential oil exhibited also an allelopathic effect against seeds of Hyoscyamus niger, as well as an insecticide effect against Sitophilus oryzae with a DL50 value of 252.4 µL/L air. These findings enhance the use of this spice as a natural food preservative and encourage its use in several fields, including pharmaceutical, cosmetics, agriculture, and therapy, that could be a strategic way to guarantee the consumer's health.

Intergenerational Transmission of Resistance of Callosobruchus maculatus to Essential Oil Treatment.

Due to the rise of numerous legal restrictions as well as the increasing emergence of resistant populations, the number of available pesticides is decreasing significantly. One of the potential alternatives often described in the literature are essential oils (EOs). However, there is a lack of research addressing the potential emergence of resistance to this group of substances. In this paper, we investigated the multi-generational effects of sublethal concentrations of rosemary oil (Rosmarinus officinalis) on physiological and biochemical parameters of the cowpea weevil (Callosobruchus maculatus) such as egg laying, hatchability, oxygen consumption and acetylcholinesterase activity. Imago, which as larvae were exposed to EO at concentrations equivalent to LC25, showed significantly lower mortality. The results obtained indicate the potential development of resistance in insects exposed to EO in concentrations corresponding to LC25. In addition, in the case of the group treated with an EO concentration corresponding to LC3.12, a stimulation effect of the above-mentioned parameters was observed, which may indicate the occurrence of a hormesis effect. The obtained results may be an important reference for the development of future guidelines and EO-based insecticides.

Persistence and ingestion characteristics of phytochemical volatiles as bio-fumigants in Sitophilus oryzae adults.

Fumigant toxicity of phytochemical volatiles has been widely reported against stored product insect pests. Such volatiles are considered as natural fumigants and bio-fumigants in post-harvest food protection research. In the present study, persistence and ingestion of diallyl disulfide, citral, eucalyptol, eugenol and menthol were investigated in Sitophilus oryzae adults in comparison with fumigant toxicity and microstructural impact in elytra. The fumigant toxicity bioassay was performed with increasing concentrations of phytochemical volatiles at 25, 125, 250 and 500 µL/L air against S. oryzae adults in 50 mL glass vials. The phytochemical residues were examined from the treated adults by Gas Chromatography coupled with Flame Ionization Detector (GC-FID) and their pathological impacts on the elytral surface was observed under Scanning Electron Microscopy (SEM). After 72 h of fumigation, diallyl disulfide and eucalyptol were identified as potential fumigants with 5.24 and 8.30 µL/L air LC50 values, respectively. GC-FID analyses showed that diallyl disulfide and eucalyptol molecules persistence (1.29 and 2.60 ppb persistence with 0.94 and 0.90 r2 values respectively at 72 h exposure) on the body surface of weevil was positively correlated with the fumigation exposure and toxicity. Whereas, phytochemical molecules ingestion into the body of weevils was not directly linked with the insect mortalities. The SEM observations indicated that diallyl disulfide and eucalyptol molecules caused severe microstructural impacts on the elytra of weevils compared to other molecules. As a result, the present study suggested that phytochemical fumigants are persisted on the body surface and caused insecticidal toxicities in S. oryzae adults. In addition, it was predicted that persisted molecules might be entered into the body of weevils via cuticular penetration.

Combined Toxicity of Cannabidiol Oil with Three Bio-Pesticides against Adults of Sitophilus Zeamais, Rhyzopertha Dominica, Prostephanus Truncatus and Trogoderma Granarium.

The present study investigates the interaction between cannabidiol (CBD) oil and three biopesticides: Azatin and two baculovirus formulations (Madex and Helicovex), both separately and in combination, in order to investigate their interaction against adults of four major coleopteran stored-product pests: Sitophilus zeamais (Coleoptera: Curculionidae), Rhyzopertha dominica (Coleoptera: Bostrichidae), Prostephanus truncatus (Coleoptera: Bostrichidae) and Trogoderma granarium (Coleoptera: Dermestidae). CBD, which has been understudied for its insecticidal properties, was applied at three different doses (500, 1500 and 3000 ppm). The biopesticides were administered at 1500 ppm. Interactions in the combined treatments were mathematically estimated as not synergistic and mostly competitive except for the combined treatments of CBD (1500 and 3000 ppm) with Azatin (1500 ppm) which were marked by an additive interaction. In its individual application, CBD oil generated the highest insect mortality while its effect was clearly dose-dependent. The findings reveal a promising effect of CBD oil against these coleopterans which had not been previously tested together.

Biological Activity of Humulus Lupulus (L.) Essential Oil and Its Main Components Against Sitophilus granarius (L.).

Besides its use in the brewing industry, hop cones appear as a powerful source of biologically active compounds, already checked for their putative anticancer, antimicrobial, and other bioactivities. Conversely, hop use in pest control remains to date under-investigated. Therefore, the biological activity of hop essential oil (EO) and its main constituents was investigated here against Sitophilus granarius. Adult contact toxicity was found 24 h after treatment with hop EO (LD50/LD90 13.30/40.23 µg/adult), and its three most abundant components, α-humulene, ß-myrcene, and ß-caryophyllene (LD50/LD90 41.87/73.51, 75.91/126.05, and 138.51/241.27 µg/adult, respectively); negligible variations at 48 h, except for α-humulene (LD50/LD90 26.83/49.49 µg/adult), were found. The fumigant toxicity of the EO and terpenes was also checked: in the absence of wheat grains, ß-myrcene showed the highest inhalation toxicity (LC50/LC90 72.78/116.92 mg/L air), whereas α-humulene, ß-caryophyllene, and the EO induced similar values (LC50/LC90 about 130/200 mg/L air); with the exception for EO, the wheat presence increased (30-50%) LC50/LC90 values. Moreover, EO and terpenes were perceived by insect antennae and elicited repellent activity. Only ß-caryophyllene showed an anticholinesterase effect, this suggesting that different mechanisms of action should be responsible for hop EO toxicity. Therefore, hop EO appears suitable for developing control means against this pest.

Synergistic Effects of Nanocomposite Films Containing Essential Oil Nanoemulsions in Combination with Ionizing Radiation for Control of Rice Weevil Sitophilus oryzae in Stored Grains.

The fumigant toxicity of eight individual essential oils (EOs; basil, cinnamon, eucalyptus, mandarin, oregano, peppermint, tea tree, and thyme) and one binary combination (thyme and oregano) for control of the rice weevil, Sitophilus oryzae, were investigated. In bioassays, all individual and combined EOs were toxic to the rice weevil. Eucalyptus EO exhibited the highest toxicity among the individual EO treatments, causing 100% mortality at a minimum concentration of 0.8 µL/mL after 24 hr of exposure. The combination treatment of oregano and thyme EO displayed higher fumigant activity than the individual oregano or thyme treatments. A stable oil-in-water nanoemulsion was evaluated using high-pressure homogenization (microfluidization [MF]) and varying the pressure and number of cycles. The droplet size of the emulsions was found to decrease from 217 to 71 nm and encapsulation efficiency increased from 37% to 84% with increasing MF pressure and number of cycles. The optimum conditions for preparing the mixture of oregano and thyme EO nanoemulsions were evaluated to be homogenization pressure of 103 MPa and three cycles. Incorporating an oregano:thyme nanoemulsion (0.75%) into cellulose nanocrystal (CNC) containing chitosan (CH/CNC), methyl cellulose (MC/CNC), and polylactic acid (PLA/CNC) composite films resulted in extended diffusion matrices causing 32% to 51% rice weevil mortality after 14 days exposure. Irradiation at 200 Gray alone caused 79% mortality and increased to 100% when combined with the bioactive chitosan film containing the oregano:thyme nanoemulsion. PRACTICAL APPLICATION: A binary combination of oregano:thyme has potential as a biopesticide against stored product pests. The encapsulation of EO nanoemulsions into biopolymeric support could be used for bioactive packaging to prevent food spoilage and extend shelf life. Combining bioactive films with irradiation can provide complete control of rice weevil in packaged rice. The system developed in this research may also be extended to explore other food-packaging films with various food models to control different types of stored pests.

Volatile Profiling, Insecticidal, Antibacterial and Antiproliferative Properties of the Essential Oils of Bursera glabrifolia Leaves.

The present article describes the chemical composition and biological activities of the essential oils of B. glabrifolia leaves obtained in four consecutive years (2015-2018). The essential oils contained α-terpineol (17.9-29.7 %), α-terpinene (12.6-17.4 %), limonene (14.9-26.8 %) and ß-pinene (2.1-16.7 %) as the most abundant volatiles. The essential oils and these volatiles showed a significant adulticide activity (p<0.01; LC50 <100 µg mL-1 ) on the maize weevil (Sitophilus zeamais) with a 100 % mortality rate within a period of 5 h. Also, the essential oils showed a strong antibacterial activity on Enterococcus faecalis ATCC 29212 (MIC, 132.7-173.6 µg mL-1 ) and Helicobacter pylori ATCC 43504 (MIC, 74.8-199.2 µg mL-1 ). The assessment of the main volatiles revealed α-terpineol as the principal antibacterial agent (MIC, 78.6-149.7 µg mL-1 ). Similarly, the essential oils exhibited a substantial in vitro antiproliferative activity on the human prostate cancer cell lines PC-3 (LC50 , 15.2-19.4 µg mL-1 ), the human ovarian carcinoma cell lines OVCAR-3 (LC50 , 27.3-53.7 µg mL-1 ) and the myelogenous leukemia cell lines K-562 (LC50 , 32.4-75.9 µg mL-1 ). α-Terpineol (LC50 , 32.4-75.9 µg mL-1 ) exhibited the strongest antiproliferative effect on these cancer cell lines (LC50 , 22.4-48.1 µg mL-1 ).

Biochemical efficacy, molecular docking and inhibitory effect of 2, 3-dimethylmaleic anhydride on insect acetylcholinesterase.

Evolution of resistance among insects to action of pesticides has led to the discovery of several insecticides (neonicotinoids and organophosphates) with new targets in insect nervous system. Present study evaluates the mode of inhibition of acetylchlonesterase (AChE), biochemical efficacy, and molecular docking of 2,3-dimethylmaleic anhydride, against Periplaneta americana and Sitophilus oryzae. The knockdown activity of 2,3-dimethylmaleic anhydride was associated with in vivo inhibition of AChE. At KD99 dosage, the 2,3-dimethylmaleic anhydride showed more than 90% inhibition of AChE activity in test insects. A significant impairment in antioxidant system was observed, characterized by alteration in superoxide dismutase and catalase activities along with increase in reduced glutathione levels. Computational docking programs provided insights in to the possible interaction between 2,3-dimethylmaleic anhydride and AChE of P. americana. Our study reveals that 2,3-dimethylmaeic anhydride elicits toxicity in S. oryzae and P. americana primarily by AChE inhibition along with oxidative stress.

Phytochemical residue profiles in rice grains fumigated with essential oils for the control of rice weevil.

In the present study, we investigated the fumigant potential of five edible essential oils (EOs) against Sitophilus oryzae and their phytochemical residues in treated grains. Among the tested EOs, peppermint oil proved significantly effective (P ≤ 0.05) on S.oryzae at 400 µl/L air concentration, inducing 83 and 100% mortalities in with-food and without-food conditions respectively over 72 h exposure. In addition, it was also observed that the binary mixtures of peppermint + lemon oil (1:1 ratio) produced an equivalent effect to that of peppermint oil alone treatments. The phytochemical residue analysis by GC-MS revealed the presence of six compounds upon 72 h exposure to EOs. Further, the analysis of physico-chemical properties of the compounds indicated a positive correlation between polar surface area (PSA) and its residual nature. The residue levels of eugenol were significantly elevated corresponding to its high PSA value (29) in clove and cinnamon oils. On the other hand, the compounds with zero PSA value imparted very less or no (D-Limonene, caryophyllene, pinene and terpinolene) residues in treated grains. With respect to the most active peppermint oil, L-menthone, menthyl acetate and eucalyptol residues were at 67, 41 and 23% levels respectively. The outcome of the present study indicate the peppermint oil as a potent fumigant against S. oryzae, and although the residues of phytochemicals in treated grains is higher; they belong to the generally recognised as safe (GRAS) status leaving no harmful effect.

Chemical constituents and insecticidal activity of essential oil of Paullinia pinnata L (Sapindaceae) / Compuestos químicos y actividad insecticida del aceite esencial de Paullinia pinnata L (Sapindaceae)

The chemical constituents and insecticidal activity of essential oil obtained by hydrodistillation of the leaves of Paullinia pinnata Linn (Sapindaceae) are being reported. The essential oil were analysed by using gas chromatography (GC) and gas chromatography coupled with mass spectrometry (GC-MS). Different concentrations (50, 100, 150, 200 and 250 mg/mL) of P. pinnata essential oil prepared separately and diluted in dimethyl sulfoxide (DMSO) were tested on the maize weevil, Sitophilus zeamais. The main constituents of the oil were pentadecanoic acid (17.9%), isoaromadendrene epoxide (11.5%) and wine lactone (11.2%). Other significant compounds of the essential oil were eremophilene (6.9%) and phytol (6.2%). The essential oil displayed 100% mortality (fumigant toxicity) against S. zeamais adults at tested concentration of 150 mg/mL with lethal concentrations (LC50) of 51.87 mg/mL air. This is the first report on the chemical constituents and insecticidal activity of essential oil of P. pinnata and may be explore as a potential natural herbal plant for the control of insect pest.

Se informan los componentes químicos y la actividad insecticida del aceite esencial obtenido por hidrodestilación de las hojas de Paullinia pinnata Linn (Sapindaceae). El aceite esencial se analizó mediante cromatografía de gases (GC) y cromatografía de gases acoplada con espectrometría de masas (GC-MS). Se ensayaron diferentes concentraciones (50, 100, 150, 200 y 250 mg/ml) de aceite esencial de P. pinnata preparado separadamente y diluido en dimetilsulfóxido (DMSO) en el gorgojo de maíz, Sitophilus zeamais. Los componentes principales del aceite fueron ácido pentadecanoico (17,9%), isoaromadendreno epóxido (11,5%) y vino lactona (11,2%). Otros compuestos significativos del aceite esencial fueron eremophilene (6,9%) y phytol (6,2%). El aceite esencial mostró una mortalidad del 100% (toxicidad fumigante) contra los adultos de S. zeamais a una concentración de 150 mg/ml con concentraciones letales (CL50) de 51,87 mg/ml de aire. Este es el primer informe sobre la composición del aceite esencial de P. pinnata y su actividad insecticida. Puede ser explorado como una potencial planta herbácea natural para el control de la plaga de insectos.

The interaction of the bioinsecticide PA1b (Pea Albumin 1 subunit b) with the insect V-ATPase triggers apoptosis.

PA1b (Pea Albumin 1, subunit b) peptide is an entomotoxin, extracted from Legume seeds, with a lethal activity towards several insect pests, such as mosquitoes, some aphids and cereal weevils. This toxin acts by binding to the subunits c and e of the plasma membrane H+-ATPase (V-ATPase) in the insect midgut. In this study, two cereal weevils, the sensitive Sitophilus oryzae strain WAA42, the resistance Sitophilus oryzae strain ISOR3 and the insensitive red flour beetle Tribolium castaneum, were used in biochemical and histological experiments to demonstrate that a PA1b/V-ATPase interaction triggers the apoptosis mechanism, resulting in insect death. Upon intoxication with PA1b, apoptotic bodies are formed in the cells of the insect midgut. In addition, caspase-3 enzyme activity occurs in the midgut of sensitive weevils after intoxication with active PA1b, but not in the midgut of resistant weevils. These biochemical data were confirmed by immuno-histochemical detection of the caspase-3 active form in the midgut of sensitive weevils. Immuno-labelling experiments also revealed that the caspase-3 active form and V-ATPase are close-localized in the insect midgut. The results concerning this unique peptidic V-ATPase inhibitor pave the way for the utilization of PA1b as a promising, more selective and eco-friendly insecticide.

Insecticidal Activity of Melaleuca alternifolia Essential Oil and RNA-Seq Analysis of Sitophilus zeamais Transcriptome in Response to Oil Fumigation.

BACKGROUND: The cereal weevil, Sitophilus zeamais is one of the most destructive pests of stored cereals worldwide. Frequent use of fumigants for managing stored-product insects has led to the development of resistance in insects. Essential oils from aromatic plants including the tea oil plant, Melaleuca alternifolia may provide environmentally friendly alternatives to currently used pest control agents. However, little is known about molecular events involved in stored-product insects in response to plant essential oil fumigation. RESULTS: M. alternifolia essential oil was shown to possess the fumigant toxicity against S. zeamais. The constituent, terpinen-4-ol was the most effective compound for fumigant toxicity. M. alternifolia essential oil significantly inhibited the activity of three enzymes in S. zeamais, including two detoxifying enzymes, glutathione S-transferase (GST), and carboxylesterase (CarE), as well as a nerve conduction enzyme, acetylcholinesterase (AChE). Comparative transcriptome analysis of S. zeamais through RNA-Seq identified a total of 3,562 differentially expressed genes (DEGs), of which 2,836 and 726 were up-regulated and down-regulated in response to M. alternifolia essential oil fumigation, respectively. Based on gene ontology (GO) analysis, the majority of DEGs were involved in insecticide detoxification and mitochondrial function. Furthermore, an abundance of DEGs mapped into the metabolism pathway in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database were associated with respiration and metabolism of xenobiotics, including cytochrome P450s, CarEs, GSTs, and ATP-binding cassette transporters (ABC transporters). Some DEGs mapped into the proteasome and phagosome pathway were found to be significantly enriched. These results led us to propose a model of insecticide action that M. alternifolia essential oil likely directly affects the hydrogen carrier to block the electron flow and interfere energy synthesis in mitochondrial respiratory chain. CONCLUSION: This is the first study to perform a comparative transcriptome analysis of S. zeamais in response to M. alternifolia essential oil fumigation. Our results provide new insights into the insecticidal mechanism of M. alternifolia essential oil fumigation against S. zeamais and eventually contribute to the management of this important agricultural pest.

Trichobaris weevils distinguish amongst toxic host plants by sensing volatiles that do not affect larval performance.

Herbivorous insects use plant metabolites to inform their host plant selection for oviposition. These host-selection behaviours are often consistent with the preference-performance hypothesis; females oviposit on hosts that maximize the performance of their offspring. However, the metabolites used for these oviposition choices and those responsible for differences in offspring performance remain unknown for ecologically relevant interactions. Here, we examined the host-selection behaviours of two sympatric weevils, the Datura (Trichobaris compacta) and tobacco (T. mucorea) weevils in field and glasshouse experiments with transgenic host plants specifically altered in different components of their secondary metabolism. Adult females of both species strongly preferred to feed on D. wrightii rather than on N. attenuata leaves, but T. mucorea preferred to oviposit on N. attenuata, while T. compacta oviposited only on D. wrightii. These oviposition behaviours increased offspring performance: T. compacta larvae only survived in D. wrightii stems and T. mucorea larvae survived better in N. attenuata than in D. wrightii stems. Choice assays with nicotine-free, JA-impaired, and sesquiterpene-over-produced isogenic N. attenuata plants revealed that although half of the T. compacta larvae survived in nicotine-free N. attenuata lines, nicotine did not influence the oviposition behaviours of both the nicotine-adapted and nicotine-sensitive species. JA-induced sesquiterpene volatiles are key compounds influencing T. mucorea females' oviposition choices, but these sesquiterpenes had no effect on larval performance. We conclude that adult females are able to choose the best host plant for their offspring and use chemicals different from those that influence larval performance to inform their oviposition decisions.

Albizia lebbeck Seed Coat Proteins Bind to Chitin and Act as a Defense against Cowpea Weevil Callosobruchus maculatus.

The seed coat is an external tissue that participates in defense against insects. In some nonhost seeds, including Albizia lebbeck, the insect Callosobruchus maculatus dies during seed coat penetration. We investigated the toxicity of A. lebbeck seed coat proteins to C. maculatus. A chitin-binding protein fraction was isolated from seed coat, and mass spectrometry showed similarity to a C1 cysteine protease. By ELM program an N-glycosylation interaction motif was identified in this protein, and by molecular docking the potential to interact with N-acetylglucosamine (NAG) was shown. The chitin-binding protein fraction was toxic to C. maculatus and was present in larval midgut and feces but not able to hydrolyze larval gut proteins. It did not interfere, though, with the intestinal cell permeability. These results indicate that the toxicity mechanism of this seed coat fraction may be related to its binding to chitin, present in the larvae gut, disturbing nutrient absorption.

Assessment of Toxicity, Antifeedant Activity, and Biochemical Responses in Stored-Grain Insects Exposed to Lethal and Sublethal Doses of Gaultheria procumbens L. Essential Oil.

The present study was undertaken to investigate the insecticidal activity of chemically characterized Gaultheria procumbens essential oil (EO) and its mode of action against the Coleopteran insects Sitophilus oryzae and Rhyzopertha dominica. Gas chromatography-mass spectrometry results depicted methyl salicylate (MS) as the major compound (96.61%) of EO. EO and its major compound methyl salicylate (MS) showed 100% mortality at 150 and 5.0 µL/L air against S. oryzae and R. dominica, respectively, on 24 h of exposure. The in vivo percent inhibition of AChE activity ranged between 6.12 and 27.50%. In addition, changes in the antioxidative defense system, superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), and oxidized glutathione (GSSG), in test insects were estimated. A significant dose-dependent response in all test parameters was observed. The results demonstrated that G. procumbens EO could play a significant role in the formulation of EO-based insecticides for the management of stored-grain insects.

Repellent Effect and Metabolite Volatile Profile of the Essential Oil of Achillea millefolium Against Aegorhinus nodipennis (Hope) (Coleoptera: Curculionidae).

Aegorhinus nodipennis (Hope) (Coleoptera: Curculionidae) is an important native pest in fruit crops that is mainly found in European hazelnut fields in the south of Chile. We investigated the behavioral response of A. nodipennis to volatile compounds released from the essential oil of Achillea millefolium and its main constituent using olfactometric bioassays. Gas chromatographic and mass spectral analysis of the A. millefolium essential oil revealed the presence of 11 compounds. Monoterpene ß-thujone (96.2%) was the main component of the oil. Other compounds identified were α-thujone, 1,8-cineole, p-cymene, and 4-terpineol, all with percentages below 1%. Both A. millefolium essential oil and thujone exhibited a repellent activity against this insect at the higher doses tested (285.7 ng/cm(2)), demonstrating their potential as repellents for this species.

Toxicity and repellency of Hoslundia opposita Vahl (Lamiaceae) leaves' essential oil against rust-red flour beetle, Tribolium castaneum Herbst (Coleoptera: Tenebrionidae).

The insecticidal properties of Hoslundia opposita Vahl (Lamiaceae) leaves' essential oil (EO) against Tribolium castaneum were investigated using contact toxicity and repellency bioassays. Mortality progressed with exposure period and ranged from 61.13% observed at 24 h after treatment (HAT) to 88.86% at 168 HAT. The LT50 (lethal time for 50% of treated adults) of H. opposita EO against T. castaneum was 10.42 h. Application of EO at 20-30 µL/30 cm(2) caused significantly (P < 0.05) higher percentage repellency than what was observed in control, 10 and 15 µL/30 cm(2) at 1 and 3 HAT, with significant repellency at 24 HAT regardless of dosage. Repellency class increased with EO dosage, with class V observed at 30 µL/30 cm(2), regardless of exposure duration. The RD50 (repellency dose for 50% of treated adults) of 15.88 and 13.37 µL/30 cm(2) for 1 and 2 HAT, respectively, was significantly higher than 0.09 µL/30 cm(2) at 24 HAT.

Insecticidal efficacy of silica gel with Juniperus oxycedrus ssp. oxycedrus (Pinales: Cupressaceae) essential oil against Sitophilus oryzae (Coleoptera: Curculionidae) and Tribolium confusum (Coleoptera: Tenebrionidae).

Laboratory bioassays were carried out to evaluate the effect of silica gel enhanced with the essential oil (EO) of Juniperus oxycedrus L. ssp. oxycedrus (Pinales: Cupressaceae) (derived from berry specimens from Greece) against adults of Sitophilus oryzae (L.) (Coleoptera: Curculionidae) and Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae). For that purpose, a dry mixture consisting of 500 mg of silica gel that had absorbed 2.18 mg of EO (total weight: 502.18 mg) was tested at three doses; 0.125, 0.250, and 0.5 g/kg of wheat, corresponding to 125, 250, and 500 ppm, respectively, and silica gel alone at 0.5 g/kg of wheat corresponding to 500 ppm, at different exposure intervals (24 and 48 h and 7 and 14 d for S. oryzae; 24 and 48 h and 7, 14, and 21 d for T. confusum). The chemical content of the specific EO was determined by gas chromatography (GC)-mass spectrometry (MS) analyses indicating the presence of 31 constituents with myrcene and germacrene-D being the predominant compounds. The bioactivity results for S. oryzae indicated that 48 h of exposure in wheat resulted in an 82% mortality for treatment with 500 ppm of the enhanced silica gel. For 7 d of exposure, 100 and 98% of S. oryzae adults died when they were treated with 500 and 250 ppm of enhanced silica gel, respectively. At 14 d of exposure, all adults died both at 250 and 500 ppm of enhanced silica gel. At 48 h, 7 and 14 d of exposure significantly less S. oryzae adults died in wheat treated with silica gel alone than at 250 or 500 ppm of enhanced silica gel. In the case of T. confusum, at 7 d of exposure, mortality in wheat treated with silica gel only was significantly higher in comparison to the other treatments. At the 14 d of exposure mortality in wheat treated with 500 ppm of silica gel alone was significantly higher than 125 and 250 ppm of the enhanced silica gel. Similar trends were also noted at 21 d of exposure, indicating that there is no enhancement effect from the addition of the EO. Results herein suggest that the simultaneous use of silica gel and J. oxycedrus ssp. oxycedrus EO enhances significantly its activity against S. oryzae.

Does cypermethrin affect enzyme activity, respiration rate and walking behavior of the maize weevil (Sitophilus zeamais)?

Insecticides cause a range of sub-lethal effects on targeted insects, which are frequently detrimental to them. However, targeted insects are able to cope with insecticides within sub-lethal ranges, which vary with their susceptibility. Here we assessed the response of three strains of the maize weevil Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae) to sub-lethal exposure to the pyrethoid insecticide cypermethrin. We expected enzyme induction associated with cypermethrin resistance since it would aid the resistant insects in surviving such exposure. Lower respiration rate and lower activity were also expected in insecticide-resistant insects since these traits are also likely to favor survivorship under insecticide exposure. Curiously though, cypermethrin did not affect activity of digestive and energy metabolism enzymes, and even reduced the activity of some enzymes (particularly for cellulase and cysteine-proteinase activity in this case). There was strain variation in response, which may be (partially) related to insecticide resistance in some strains. Sub-lethal exposure to cypermethrin depressed proteolytic and mainly cellulolytic activity in the exposed insects, which is likely to impair their fitness. However, such exposure did not affect respiration rate and walking behavior of the insects (except for the susceptible strain where walking activity was reduced). Walking activity varies with strain and may minimize insecticide exposure, which should be a concern, particularly if associated with (physiological) insecticide resistance.