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.

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.

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.

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.

Evaluation of the toxicity of 17 essential oils against Choristoneura rosaceana (Lepidoptera: Tortricidae) and Trichoplusia ni (Lepidoptera: Noctuidae).

BACKGROUND: The obliquebanded leafroller, Choristoneura rosaceana Harris, and the cabbage looper, Trichoplusia ni Hübner, are serious fruit and vegetable pests requiring multiple insecticide applications per year. To reduce non-target exposure to hazardous insecticides and to curb resistance development, alternative controls are required. Accordingly, a selection of 17 essential oils was screened against both lepidopteran pests, and the influence of azinphos-methyl resistance on essential oil toxicity to C. rosaceana was studied. RESULTS: Of the 17 essential oils screened, patchouli oil (Pogostemon cablin Benth.) and thyme oil (Thymus vulgaris L.) were selected for further testing against C. rosaceana, whereas patchouli oil, garlic oil (Allium sativum L.) and lemongrass oil (Cymopogon nardus L.) were selected for further testing against T. ni. LC(50) and LD(50) values confirmed that patchouli oil was the most toxic to C. rosaceana larvae, with LC(50) = 2.8 µL mL(-1) and LD(50) = 8.0 µg insect(-1). Garlic oil was the most toxic oil to T. ni larvae with LC(50) = 3.3 µL mL(-1) and LD(50) = 22.7 µg insect(-1), followed by patchouli oil and lemongrass oil. Azinphos-methyl-resistant leafrollers were 1.5-fold more tolerant to patchouli oil and 2.0-fold more tolerant to thyme oil. CONCLUSIONS: Based on these results, patchouli oil and other essential oils have sufficient efficacy to be considered as components of an essential oil-based insecticide that targets these lepidopteran pests.

Comparative toxicity of essential oils of Litsea pungens and Litsea cubeba and blends of their major constituents against the cabbage looper, Trichoplusia ni.

Contact toxicity of essential oils of Litsea pungens Hemsl. and L. cubeba (Lour.) Pers. (Lauraceae) and of blends of their major constituents was assessed against third-instar Trichoplusia ni larvae via topical application. Both oils showed moderate activity against T. ni larvae with LD(50) values of 87.1 and 112.5 microg/larva, respectively. 1,8-Cineole from the essential oil of L. pungens and gamma-terpinene from the oil of L. cubeba accounted for much of the toxicity of the oils to T. ni larvae. The toxicity of blends of selected constituents indicated a synergistic effect among putatively active and inactive constituents, with the presence of all constituents necessary for full toxicity of the natural oils. The results show that essential oils of L. pungens and L. cubeba and some of their constituents have potential for development as botanical insecticides.

Green peach aphid, Myzus persicae (Hemiptera: Aphididae), reproduction during exposure to sublethal concentrations of imidacloprid and azadirachtin.

BACKGROUND: Resurgence of insect pests following insecticide applications is often attributed to natural enemy disturbance, but hormesis could be an alternative or additional mechanism. Green peach aphid, Myzus persicae (Sulzer), is an important insect pest of many crops worldwide that may be exposed to sublethal insecticide concentrations over time. Here, the hypothesis that exposure to low concentrations of imidacloprid and azadirachtin can induce hormetic responses in M. persicae is tested in the laboratory. RESULTS: When insects were exposed to potato leaf discs dipped in sublethal concentrations of insecticide, almost all measured endpoints-adult longevity, F1 production, F1 survival and F2 production-were affected, and a statistically significant (P < 0.05) stimulatory response was recorded for F2 production following exposure to imidacloprid. No other measures for hormesis were statistically significant, but other trends of hormetic response were consistently observed. CONCLUSIONS: Given that variable distribution and degradation of insecticides in the field would result in a wide range of concentrations over time and space, these laboratory experiments suggest that exposure to sublethal concentrations of imidacloprid and azadirachtin could stimulate reproduction in M. persicae.

Botanical insecticides: for richer, for poorer.

Botanical insecticides presently play only a minor role in insect pest management and crop protection; increasingly stringent regulatory requirements in many jurisdictions have prevented all but a handful of botanical products from reaching the marketplace in North America and Europe in the past 20 years. Nonetheless, the regulatory environment and public health needs are creating opportunities for the use of botanicals in industrialized countries in situations where human and animal health are foremost--for pest control in and around homes and gardens, in commercial kitchens and food storage facilities and on companion animals. Botanicals may also find favour in organic food production, both in the field and in controlled environments. In this review it is argued that the greatest benefits from botanicals might be achieved in developing countries, where human pesticide poisonings are most prevalent. Recent studies in Africa suggest that extracts of locally available plants can be effective as crop protectants, either used alone or in mixtures with conventional insecticides at reduced rates. These studies suggest that indigenous knowledge and traditional practice can make valuable contributions to domestic food production in countries where strict enforcement of pesticide regulations is impractical.

Comparative toxicity of Rosmarinus officinalis L. essential oil and blends of its major constituents against Tetranychus urticae Koch (Acari: Tetranychidae) on two different host plants.

Bioassays of Rosmarinus officinalis L. essential oil and blends of its major constituents were conducted using host-specific strains of the two-spotted spider mite, Tetranychus urticae Koch, on bean and tomato plants. Two constituents tested individually against a bean host strain and five constituents tested individually against a tomato host strain accounted for most of the toxicity of the natural oil. Other constituents were relatively inactive when tested individually. Toxicity of blends of selected constituents indicated a synergistic effect among the active and inactive constituents, with the presence of all constituents necessary to equal the toxicity of the natural oil.

Botanical insecticides, deterrents, and repellents in modern agriculture and an increasingly regulated world.

Botanical insecticides have long been touted as attractive alternatives to synthetic chemical insecticides for pest management because botanicals reputedly pose little threat to the environment or to human health. The body of scientific literature documenting bioactivity of plant derivatives to arthropod pests continues to expand, yet only a handful of botanicals are currently used in agriculture in the industrialized world, and there are few prospects for commercial development of new botanical products. Pyrethrum and neem are well established commercially, pesticides based on plant essential oils have recently entered the marketplace, and the use of rotenone appears to be waning. A number of plant substances have been considered for use as insect antifeedants or repellents, but apart from some natural mosquito repellents, little commercial success has ensued for plant substances that modify arthropod behavior. Several factors appear to limit the success of botanicals, most notably regulatory barriers and the availability of competing products (newer synthetics, fermentation products, microbials) that are cost-effective and relatively safe compared with their predecessors. In the context of agricultural pest management, botanical insecticides are best suited for use in organic food production in industrialized countries but can play a much greater role in the production and postharvest protection of food in developing countries.

Efficacy and persistence of rosemary oil as an acaricide against twospotted spider mite (Acari: Tetranychidae) on greenhouse tomato.

Efficacy of rosemary, Rosmarinus officinalis L., essential oil was assessed against twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), as well as effects on the tomato, Lycopersicum esculatum Mill., host plant and biocontrol agents. Laboratory bioassay results indicated that pure rosemary oil and EcoTrol (a rosemary oil-based pesticide) caused complete mortality of spider mites at concentrations that are not phytotoxic to the host plant. The predatory mite Phytoseiulus persimilis Athias-Henriot is less susceptible to rosemary oil and EcoTrol than twospotted spider mite both in the laboratory and the greenhouse. Rosemary oil repels spider mites and can affect oviposition behavior. Moreover, rosemary oil and rosemary oil-based pesticides are nonpersistent in the environment, and their lethal and sublethal effects fade within 1 or 2 d. EcoTrol is safe to tomato foliage, flowers, and fruit even at double the recommended label rate. A greenhouse trial indicated that a single application of EcoTrol at its recommended label rate could reduce a twospotted spider mite population by 52%. At that rate, EcoTrol did not cause any mortality in P. persimilis nor did it affect their eggs. In general, EcoTrol was found to be a suitable option for small-scale integrated pest management programs for controlling twospotted spider mites on greenhouse tomato plants.

Insecticidal activity of the essential oil of Ligusticum mutellina roots.

The essential oil obtained from roots of different collections of Ligusticum mutellina was tested against 3rd instar armyworms, Pseudaletia unipuncta (Lepidoptera: Noctuidae), for insecticidal activity. The main compounds were isolated and their structures were elucidated using 2D-NMR techniques. Our collections contained dillapiole, ligustilide and myristicin as major compounds. The previously reported sarisan was not present, moreover its occurrence in L. mutellina should be revised based on our findings.

Generalization of a habituated feeding deterrent response to unrelated antifeedants following prolonged exposure in a generalist herbivore, Trichoplusia ni.

The possibility of generalization of habituated response to unrelated feeding deterrents following prolonged exposure was examined in third instar Trichoplusia ni (Lepidoptera: Noctuidae) larvae by rearing them on antifeedants and then testing with other unrelated antifeedants. We introduced neonate larvae (< 24-hr old) onto cabbage leaves treated with crude seed extracts of Melia volkensii (Meliaceae) or oil of Origanum vulgare ("oregano") (Lamiaceae) and allowed them to feed until early in the third instar. Naive larvae were reared on cabbage leaves treated with carrier solvent alone. Both experienced and naïve larvae were tested for feeding deterrent response with the same and the different extracts in a leaf disc choice bioassay. Habituation was generalized to both M. volkensii and oregano following prolonged exposure to either plant extract and also to a pure allelochemical, thymol, following prolonged exposure to either digitoxin or xanthotoxin. However, there was no generalization of the habituated response to oregano following prolonged exposure to digitoxin or thymol, or to thymol or xanthotoxin following prolonged exposure to oregano or M. volkensii. Our results demonstrate that habituated response to feeding deterrents in a polyphagous insect herbivore can be generalized among and between plant extracts and pure allelochemicals, but not in all situations. The implications of such behavioral plasticity in herbivorous insects for the use of antifeedants as crop protectants or for host plant shifts is discussed.

Larval exposure to oviposition deterrents alters subsequent oviposition behavior in generalist, Trichoplusia ni and specialist, Plutella xylostella moths.

The present study was undertaken to determine the effects of larval feeding experience on subsequent oviposition behavior of the resulting moths. Larvae of the cabbage looper (Trichoplusia ni, Noctuidae) and the diamondback moth (Plutella xylostella, Plutellidae) were exposed to the phenylpropanoid allelochemical trans-anethole (at 100 ppm fw in artificial diet) or the limonoid allelochemical toosendanin (10 ppm sprayed on cabbage leaves). Both compounds had been shown to deter oviposition in naive moths in previous choice tests. Moths developing from "experienced" larvae (both sexes) showed a decrease in oviposition deterrence response when given a choice between control and treated leaves, unlike naïve moths. This phenomenon, analogous to habituation to feeding deterrents in lepidopteran larva, occurred irrespective of duration of feeding on the deterrent compound. We also observed that F1 larvae resulting from experienced moths (previously exposed to toosendanin as larvae) grew as well on toosendanin-treated foliage as on control foliage. In contrast, growth of F1 larvae from naïve moths was significantly impaired by toosendanin. These results demonstrate that host-selection behavior in cabbage looper (a generalist) and diamondback moth (a specialist) may be shaped by feeding experience according to Hopkins' Host Selection Principle in addition to chemical legacy.