Insecticidal Activity of 28 Essential Oils and a Commercial Product Containing Cinnamomum cassia Bark Essential Oil against Sitophilus zeamais Motschulsky.

Maize weevils, Sitophilus zeamais, are stored product pests mostly found in warm and humid regions around the globe. In the present study, acute toxicity via contact and residual bioassay and fumigant bioassay of 28 essential oils as well as their attraction-inhibitory activity against the adults of S. zeamais were evaluated. Chemical composition of the essential oils was analyzed by gas chromatography-mass spectrometry, and a compound elimination assay was conducted on the four most active oils (cinnamon, tea tree, ylang ylang, and marjoram oils) to identify major active constituents. Amongst the oils examined, cinnamon oil was the most active in both contact/residual and fumigant bioassays, and exhibited strong behavioral inhibitory activity. Based on the compound elimination assay and chemical analyses, trans-cinnamaldehyde in cinnamon oil, and terpinen-4-ol in tea tree and marjoram oils were identified as the major active components. Although cinnamon oil seemed promising in the lab-scale bioassay without rice grains, it failed to exhibit strong insecticidal activity when the container was filled with rice. When a cinnamon oil-based product was applied both in an empty glass jar and a rice-filled container, all weevils in the empty jar were killed, whereas fewer than 15% died in the rice-filled container.

Variation among 532 genomes unveils the origin and evolutionary history of a global insect herbivore.

The diamondback moth, Plutella xylostella is a cosmopolitan pest that has evolved resistance to all classes of insecticide, and costs the world economy an estimated US $4-5 billion annually. We analyse patterns of variation among 532 P. xylostella genomes, representing a worldwide sample of 114 populations. We find evidence that suggests South America is the geographical area of origin of this species, challenging earlier hypotheses of an Old-World origin. Our analysis indicates that Plutella xylostella has experienced three major expansions across the world, mainly facilitated by European colonization and global trade. We identify genomic signatures of selection in genes related to metabolic and signaling pathways that could be evidence of environmental adaptation. This evolutionary history of P. xylostella provides insights into transoceanic movements that have enabled it to become a worldwide pest.

Bioinsecticides based on plant essential oils: a short overview.

Interest in the discovery and development of plant essential oils for use as bioinsecticides has grown enormously in the past 20 years. However, successful commercialization and utilization of crop protection products based on essential oils has thus far lagged far behind their promise based on this large body of research, most notably because with the exceptions of the USA and Australia, such products receive no special status from regulatory agencies that approve new pesticides for use. Essential oil-based insecticides have now been used in the USA for well over a decade, and more recently have seen use in the European Union (EU), Korea, and about a dozen other countries, with demonstrated efficacy against a wide range of pests and in numerous crop systems. For the most part these products are based on commodity essential oils developed as flavor and fragrance agents for the food and cosmetic industries, as there are formidable logistic, economic, and regulatory challenges to the use of many other essential oils that otherwise possess potentially useful bioactivity against pests. In spite of these limitations, the overall prospects for biopesticides, including those based on essential oils, are encouraging as the demand for sustainably-produced and/or organic food continues to increase worldwide.

Botanical Insecticides in the Twenty-First Century-Fulfilling Their Promise?

Academic interest in plant natural products with insecticidal properties has continued to grow in the past 20 years, while commercialization of new botanical insecticides and market expansion of existing botanicals has lagged considerably behind. Insecticides based on pyrethrum and neem (azadirachtin) continue to be standard bearers in this class of pesticides, but globally, their increased presence is largely a consequence of introduction into new jurisdictions. Insecticides based on plant essential oils are just beginning to emerge as useful plant protectants. Some countries (such as Turkey, Uruguay, the United Arab Emirates, and Australia) have relaxed regulatory requirements for specific plant extracts and oils, while in North America and the European Union, stricter requirements have slowed progress toward commercialization of new products. Botanicals are likely to remain niche products in many agricultural regions and may have the greatest impact in developing countries in tropical regions where the source plants are readily available and conventional products are both expensive and dangerous to users.

Oliveria decumbens, a Bioactive Essential Oil: Chemical Composition and Biological Activities.

Oliveria decumbens is an aromatic plant traditionally used for treatment of infections and gastrointestinal diseases. In the present study, the volatile oil of the plant was obtained by hydrodistillation and analyzed by GC-MS. In addition, antibacterial and anti-Helicobacter pylori activities of this essential oil were determined using disc diffusion and agar dilution methods, respectively. Insecticidal activity was assessed through topical and fumigation application of the essential oil to cabbage looper larvae. Acetylcholinesterase (AChE) inhibition by the essential oil was examined using Ellman's method. Furthermore, its cytotoxic potential against three different cancer cell lines was assessed using the MTT assay. The phenolic monoterpenoids, thymol (38.79%), and carvacrol (36.30%) were identified as major constituents of the essential oil. We observed significant antibacterial activity of the essential oil against H. pylori (MIC=20.4 µg /mL) as well as other tested bacteria, except for Pseudomonas aeruginosa. O. decumbens essential oil showed significant toxicity to cabbage looper larvae with LD50 value of 52.1 µg /larva following topical and fumigant administration. O. decumbens essential oil was considerably inhibitory to acetylcholinesterase activity (IC50 = 0.117 µg/mL). Cytotoxic assay of the volatile oil resulted in IC50 = 0.065, 0.104, and 0.141 µg/mL for MCF-7, T47D and MDA- MB-231 cell lines, respectively. According to our data, this species with high concentrations of thymol and carvacrol could be considered as a natural source for pharmaceutical products.

Effects of rosemary, thyme and lemongrass oils and their major constituents on detoxifying enzyme activity and insecticidal activity in Trichoplusia ni.

Although there have been many reports on the synergistic interactions among the major constituents of plant essential oils regarding insecticidal activity, their underlying mechanism of synergy is poorly understood. In our previous studies, we found each of the two most abundant constituents of rosemary (Rosmarinus officinalis L.), thyme (Thymus vulgaris L.) and lemongrass (Cymbopogon citratus Stapf.) essential oils can be synergistic against the larvae of the cabbage looper, Trichoplusia ni at their natural proportion or equivalent blending ratios. In the present study, we investigated whether the enhanced toxicity between the major constituents could be the result of inhibited enzyme activity of cytochrome P450s, general esterases or glutathione S-transferases which are highly related to the development of insecticide resistance. Overall, although some combinations showed mild inhibitory activity, at least for these essential oils and their major constituents, inhibition of detoxication enzyme activity is unlikely to be a direct cause of increased toxicity in the cabbage looper. The results point to other factors, such as multiple modes-of-action or enhanced penetration through the cuticular layer, playing important roles in the elevated insecticidal activity. Moreover, application of enzyme inhibitors sometimes resulted in decreased activity when mixed with the target compounds, but these antagonistic interactions disappeared when they were applied separately, suggesting that the enzyme inhibitors can sometimes influence the penetrations of toxicants.

Penetration-enhancement underlies synergy of plant essential oil terpenoids as insecticides in the cabbage looper, Trichoplusia ni.

Many plant essential oils and their terpenoid constituents possess bioactivities including insecticidal activity, and they sometimes act synergistically when mixed. Although several hypotheses for this have been proposed, the underlying mechanism has not been fully elucidated thus far. In the present study, we report that in larvae of the cabbage looper, Trichoplusia ni, most synergistic or antagonistic insecticidal activities among mixtures of plant essential oil constituents are pharmacokinetic effects, owing to changes in solubility as well as spreadability on a wax layer. Among the major constituents of rosemary (Rosmarinus officinalis) oil, in vitro analysis revealed up to a 19-fold increase in penetration of camphor in a binary mixture with 1,8-cineole through the larval integument, suggesting increased penetration as the major mechanism for synergy. A total of 138 synergistic or antagonistic interactions among 39 compounds were identified in binary mixtures via topical application, and these were highly correlated to changes in surface tension as measured by contact angle of the mixtures on a beeswax layer. Among compounds tested, trans-anethole alone showed evidence of internal synergy, whereas most of remaining synergistic or antagonistic combinations among the three most active compounds were identified as penetration-related interactions, confirmed via a divided-application bioassay.

Turmeric powder and its derivatives from Curcuma longa rhizomes: Insecticidal effects on cabbage looper and the role of synergists.

Curcuma longa has well-known insecticidal and repellent effects on insect pests, but its impact on Trichoplusia ni is unknown. In this study, the compound ar-turmerone, extracted and purified from C. longa rhizomes, was identified, and its insecticidal effects, along with turmeric powder, curcuminoid pigments and crude essential oil were evaluated against this important agricultural pest. The role of natural (sesamol and piperonal) and synthetic [piperonyl butoxide (PBO)] synergists under laboratory and greenhouse conditions were also evaluated. The concentration of ar-turmerone in C. longa rhizomes harvested was 0.32% (dwt). Turmeric powder and its derivatives caused 10-20% mortality in third instar T. ni at a very low dose (10 µg/larva). Addition of PBO increased toxicity of turmeric powder and its derivatives (90-97% mortality) in most binary combinations (5 µg of turmeric powder or its derivatives +5 µg of PBO), but neither piperonal nor sesamol were active as synergists. The compound ar-turmerone alone and the combination with PBO reduced larval weight on treated Brassica oleracea in the laboratory and in greenhouse experiments, compared with the negative control. The compound ar-turmerone could be used as a low cost botanical insecticide for integrated management of cabbage looper in vegetable production.

Metabolism of citral, the major constituent of lemongrass oil, in the cabbage looper, Trichoplusia ni, and effects of enzyme inhibitors on toxicity and metabolism.

Although screening for new and reliable sources of botanical insecticides remains important, finding ways to improve the efficacy of those already in use through better understanding of their modes-of-action or metabolic pathways, or by improving formulations, deserves greater attention as the latter may present lesser regulation hurdles. Metabolic processing of citral (a combination of the stereoisomers geranial and neral), a main constituent of lemongrass (Cymbopogon citratus) essential oil has not been previously examined in insects. To address this, we investigated insecticidal activities of lemongrass oil and citral, as well as the metabolism of citral in larvae of the cabbage looper, Trichoplusia ni, in associations with well-known enzyme inhibitors. Among the inhibitors tested, piperonyl butoxide showed the highest increase in toxicity followed by triphenyl phosphate, but no synergistic interaction between the inhibitors was observed. Topical application of citral to fifth instar larvae produced mild reductions in food consumption, and frass analysis after 24h revealed geranic acid (99.7%) and neric acid (98.8%) as major metabolites of citral. Neither citral nor any other metabolites were found following in vivo analysis of larvae after 24h, and no significant effect of enzyme inhibitors was observed on diet consumption or citral metabolism.

Identification of a Chrysanthemic Ester as an Apolipoprotein E Inducer in Astrocytes.

The apolipoprotein E (APOE) gene is the most highly associated susceptibility locus for late onset Alzheimer's Disease (AD), and augmenting the beneficial physiological functions of apoE is a proposed therapeutic strategy. In a high throughput phenotypic screen for small molecules that enhance apoE secretion from human CCF-STTG1 astrocytoma cells, we show the chrysanthemic ester 82879 robustly increases expressed apoE up to 9.4-fold and secreted apoE up to 6-fold and is associated with increased total cholesterol in conditioned media. Compound 82879 is unique as structural analogues, including pyrethroid esters, show no effect on apoE expression or secretion. 82879 also stimulates liver x receptor (LXR) target genes including ATP binding cassette A1 (ABCA1), LXRα and inducible degrader of low density lipoprotein receptor (IDOL) at both mRNA and protein levels. In particular, the lipid transporter ABCA1 was increased by up to 10.6-fold upon 82879 treatment. The findings from CCF-STTG1 cells were confirmed in primary human astrocytes from three donors, where increased apoE and ABCA1 was observed along with elevated secretion of high-density lipoprotein (HDL)-like apoE particles. Nuclear receptor transactivation assays revealed modest direct LXR agonism by compound 82879, yet 10 µM of 82879 significantly upregulated apoE mRNA in mouse embryonic fibroblasts (MEFs) depleted of both LXRα and LXRß, demonstrating that 82879 can also induce apoE expression independent of LXR transactivation. By contrast, deletion of LXRs in MEFs completely blocked mRNA changes in ABCA1 even at 10 µM of 82879, indicating the ability of 82879 to stimulate ABCA1 expression is entirely dependent on LXR transactivation. Taken together, compound 82879 is a novel chrysanthemic ester capable of modulating apoE secretion as well as apoE-associated lipid metabolic pathways in astrocytes, which is structurally and mechanistically distinct from known LXR agonists.

Comparative and synergistic activity of Rosmarinus officinalis L. essential oil constituents against the larvae and an ovarian cell line of the cabbage looper, Trichoplusia ni (Lepidoptera: Noctuidae).

BACKGROUND: Plant essential oils are usually complex mixtures, and many factors can affect their chemical composition. To identify relationships between the composition and bioactivity of the constituents, comparative and synergistic interactions of the major constituents of rosemary essential oil were evaluated against third-instar larvae and an ovarian cell line of the cabbage looper, Trichoplusia ni, via different methods of application. RESULTS: The major constituents of the rosemary oil we used were 1,8-cineole, (±)-camphor, (+)-α-pinene and camphene. Via topical application to larvae, 1,8-cineole was identified as the major active compound, whereas via fumigation, 1,8-cineole and (±)-camphor, and in a cytotoxicity assay, (+)-α-pinene, were determined to be the major active principles. Several combinations of these constituents exhibited synergistic insecticidal activities when topically applied, particularly among combinations of three major constituents, (±)-camphor, (+)-α-pinene and camphene. A binary mixture of 1,8-cineole and (±)-camphor showed enhanced activity, with a synergy ratio of 1.72. CONCLUSION: Based on our results, the insecticidal activity of rosemary oil appears to be a consequence of the synergistic interaction between 1,8-cineole and (±)-camphor, and (±)-camphor should be considered a promising synergizing agent.

A renaissance for botanical insecticides?

Botanical insecticides continue to be a subject of keen interest among the international research community, reflected in the steady growth in scientific publications devoted to the subject. Until very recently though, the translation of that theory to practice, i.e. the commercialisation and adoption of new botanical insecticides in the marketplace, has seriously lagged behind. Strict regulatory regimes, long the bane of small pesticide producers, are beginning to relax some of the data requirements for 'low-risk' pesticide products, facilitating movement of more botanicals into the commercial arena. In this paper I discuss some of the jurisdictions where botanicals are increasingly finding favour, some of the newer botanical insecticides in the plant and animal health arsenal and some of the specific sectors where botanicals are most likely to compete effectively with other types of insecticidal product.

Enhanced cuticular penetration as the mechanism for synergy of insecticidal constituents of rosemary essential oil in Trichoplusia ni.

Synergistic interactions between constituents of essential oils have been reported for several areas of research. In the present study, mechanisms that could explain the synergistic action of the two major insecticidal constituents of rosemary oil, 1,8-cineole and camphor against the cabbage looper, Trichoplusia ni were investigated. 1,8-Cineole was more toxic than camphor when applied topically to larvae, and when coadministered in their ratio naturally occurring in rosemary oil, the binary mixture was synergistic. However, when injected directly into larvae, camphor was more toxic than 1,8-cineole. GC-MS analyses showed that penetration of topically-applied camphor was significantly enhanced when it was mixed with 1,8-cineole in the natural ratio. A bioassay combining injection and topical application methods confirmed the increased penetration of both compounds when mixed, showing the same bioactivity as seen for higher amounts applied individually. Lowered surface tension as well as increased solubility of camphor by 1,8-cineole, along with the interaction between 1,8-cineole and the lipid layer of the insect's cuticle may explain the enhanced penetration of camphor. Because of the similarities in biological function of animal and microbial membranes, our finding has potential for application in other fields of study.

Dhilirolides E-N, meroterpenoids produced in culture by the fungus Penicillium purpurogenum collected in Sri Lanka: structure elucidation, stable isotope feeding studies, and insecticidal activity.

Extracts of laboratory cultures of the fungus Penicilium purpurogenum obtained from rotting fruit of the tree Averrhoa bilimbi growing in Sri Lanka have yielded 10 new meroterpenoids, dhilirolides E-N (5-14). The structures of the new dhilirolides have been elucidated by analysis of spectroscopic data and a single-crystal X-ray diffraction analysis of dhilirolide L (12). Dhilirolides A-N (1-14) represent the four unprecedented and rearranged dhilirane, isodhilirane, 14,15-dinordhilirane, and 23,24-dinorisodhilirane meroterpenoid carbon skeletons. Stable isotope feeding studies have confirmed the meroterpenoid biogenetic origin of the dhilirolides and provided support for a proposed genesis of the new carbon skeletons. Dhilirolide L (12) showed significant feeding inhibition and sublethal developmental disruption in the cabbage looper Trichoplusia ni, an important agricultural pest, at low concentrations.

Botanical insecticides inspired by plant-herbivore chemical interactions.

Plants have evolved a plethora of secondary chemicals to protect themselves against herbivores and pathogens, some of which have been used historically for pest management. The extraction methods used by industry render many phytochemicals ineffective as insecticides despite their bioactivity in the natural context. In this review, we examine how plants use their secondary chemicals in nature and compare this with how they are used as insecticides to understand why the efficacy of botanical insecticides can be so variable. If the commercial production of botanical insecticides is to become a viable pest management option, factors such as production cost, resource availability, and extraction and formulation techniques need be considered alongside innovative application technologies to ensure consistent efficacy of botanical insecticides.

Botanical insecticide research: many publications, limited useful data.

Our analysis of >20000 papers on botanical insecticides from 1980 to 2012, indicates major growth in the number of papers published annually (61 in 1980 to 1207 in 2012), and their proportion among all papers on insecticides (1.43% in 1980 to 21.38% in 2012). However, only one-third of 197 random articles among the 1086 papers on botanical insecticides published in 2011 included any chemical data or characterization; and only a quarter of them included positive controls. Therefore, a substantial portion of recently published studies has design flaws that limit reproducibility and comparisons with other and/or future studies. In our opinion, much of the scientific literature on this subject is of limited use in the progress toward commercialization or advancement of knowledge, given the resources expended.

Horizontal transfer of diatomaceous earth and botanical insecticides in the common bed bug, Cimex lectularius L.; hemiptera: cimicidae.

BACKGROUND: Horizontal transfer of insecticide occurs when insects contact or ingest an insecticide, return to an aggregation or a nest, and transfer the insecticide to other conspecific insects through contact. This phenomenon has been reported in a number of insects including social insects, however it has not been reported in bed bugs. Since horizontal transfer can facilitate the spread of insecticide into hard to reach spaces, it could contribute greatly to the management of these public health pests. METHODOLOGY/RESULTS: To demonstrate horizontal transfer of diatomaceous earth and botanical insecticides in C. lectularius, an exposed (donor) bed bug, following a 10-minute acquisition period, was placed with unexposed (recipient) bed bugs. Mortality data clearly demonstrates that diatomaceous earth (DE 51) was actively transferred from a single exposed bug to unexposed bugs in a concentration dependent manner. LC50 values varied from 24.4 mg at 48 h to 5.1 mg at 216 h when a single exposed bed bug was placed with 5 unexposed bed bugs. LT50 values also exhibited a concentration response. LT50 values varied from 1.8 days to 8.4 days when a 'donor' bug exposed to 20 and 5 mg of dust respectively was placed with 5 'recipient' bugs. Dust was also actively transferred from adult bed bugs to the nymphs. In addition we observed horizontal transfer of botanical insecticides including neem, ryania, and rotenone to varying degrees. CONCLUSION/SIGNIFICANCE: Our data clearly demonstrate horizontal transfer of diatomaceous earth and botanical insecticides in the common bed bug, C. lectularius. Use of a fluorescent dust provided visual confirmation that contaminated bed bugs transfer dust to untreated bed bugs in harborage. This result is important because bedbugs live in hard-to-reach places and interaction between conspecifics can be exploited for delivery and dissemination of management products directed at this public health pest.

Population density and feeding duration of cabbage looper larvae on tomato plants alter the levels of plant volatile emissions.

BACKGROUND: As part of their indirect defense, plants under herbivore attack release volatile chemicals that attract natural enemies of the herbivore. This is a very well-documented phenomenon. However, relatively few studies have investigated the response of plants to different population levels of herbivores and their feeding duration. RESULTS: Working with larvae of the cabbage looper, Trichoplusia ni (Hübner), and tomato plants, Lycopersicon esculentum Mill cv. clarence, and using an ultrafast gas chromatograph (the zNose™) for volatile analyses, the authors studied the effect of larval density and feeding duration on levels of plant volatile emissions. Intense herbivory caused higher emission levels of the herbivore-induced plant volatiles (HIPVs) (Z)-3-hexenyl acetate, (E)-ß-ocimene and ß-caryophyllene than those caused by moderate herbivory. When herbivory had ceased following 12-24 h of larval feeding, plants kept releasing HIPVs at a high level for a longer period of time than they did following only 6 h of larval feeding. The plants' slow adjustment in their volatile emissions following prolonged larval feeding might be strategic, as such feeding is more likely to have ceased just temporarily. CONCLUSION: This information may help in the development of a pest monitoring system that is based on herbivore-induced plant volatiles.

Trapping whiteflies? A comparison of greenhouse whitefly (Trialeurodes vaporariorum) responses to trap crops and yellow sticky traps.

BACKGROUND: Greenhouse whiteflies, Trialeurodes vaporariorum (Westwood), are notorious pests that, through the reduction of crop yields and excretion of honeydew, cause significant economic losses for sweet pepper (Capsicum annuum L.) growers. Chemical and biological controls are the two most common forms of whitefly management in greenhouses. Consequently, insecticide resistance and inadequate control have rejuvenated interest in alternative tactics. In the present study, whitefly responses to trap crops (eggplant and squash) and yellow sticky traps were compared in order to identify the most effective traps for dispersing and resident adults. RESULTS: Results indicated that yellow sticky traps were most effective at trapping adult whiteflies. Significantly more dispersing whiteflies were recorded on eggplant than on squash trap crops. None of the traps significantly reduced adult populations on the main crops (peppers) compared with the control. However, yellow sticky traps did significantly reduce oviposition on peppers. CONCLUSIONS: Adult whiteflies were most effectively trapped on yellow sticky traps followed by eggplant trap crops. Further study of whitefly trapping using visual cues may enhance trapping management. Specifically, research combining yellow sticky traps with other control strategies is recommended.

Dishabituating long-term memory for gustatory habituation in the cabbage looper, Trichoplusia ni.

The gustatory rejection response of the cabbage looper, Trichoplusia ni (Lepidoptera: Noctuidae), habituates to antifeedant compounds, allowing for the consumption of deterrent yet nontoxic plant materials. In the present study, we demonstrate that habituation to an antifeedant compound (quinine) persists through the moult between larval instars. As an indirect test of whether the memory was protein synthesis-dependent, we tested whether disrupting protein synthesis would block memory reconsolidation after a reminder. The results indicated that disrupting protein synthesis in habituated larvae following a reminder treatment (reexposure to quinine) eliminated the memory for habituation and restored the antifeedant properties of the quinine. We then examined whether the learned memory could be dishabituated and whether this would disrupt or eliminate long-term memory. We show that 6 hr after exposure to a novel/noxious stimulus (a second antifeedant, xanthotoxin) habituated larvae showed a transient dishabituation-like effect in which the quinine deterred feeding again. However, this effect did not permanently eliminate the habituation produced by the extended exposure as larvae tested 72 hr after xanthotoxin exposure again showed a willingness to consume the quinine treated leaves, indicating that the earlier habituation was still present.