Chemical Cues From Honeydew and Cuticular Extracts of Trialeurodes Vaporariorum Serve as Kairomones for The Parasitoid Encarsia Formosa.

Kairomones are semiochemicals that are emitted by an organism and which mediate interspecific interaction that is of benefit to an organism of another species that receives these chemical substances. Parasitoids find and recognize their hosts through eavesdropping on the kairomones emitted from the by-products or the body of the host. Hemipteran insect pests feed on plant sap and excrete the digested plant materials as honeydew. Honeydew serves as a nutritional food source for parasitoids and a medium for micro-organisms whose activity induces the release of volatiles exploited by parasitoids for host location. The parasitoid Encarsia formosa preferentially parasitizes its host, the greenhouse whitefly, Trialeurodes vaporariorum, on tomato Solanum lycopersicum, but little is known about the chemicals that mediate these interactions. We investigated the olfactory responses of the parasitoid E. formosa to odours from honeydew and nymphs of T. vaporariorum in a Y-tube olfactometer. Arrestment behaviour of the parasitoid to honeydew and nymph extracts, as well as to synthetic hydrocarbons, was also observed in Petri-dish bioassays. We found that T. vaporariorum honeydew volatiles attracted the parasitoid E. formosa but odours from the whitefly nymphs did not. We also found that the parasitoid spent more time searching on areas treated with extracts of honeydew and nymphs than on untreated areas. Gas-chromatography-mass spectrometric analysis revealed that the honeydew volatiles contained compounds such as (Z)-3-hexenol, δ-3-carene, 3-octanone, α-phellandrene, methyl salicylate, ß-ocimene, ß-myrcene, and (E)-ß-caryophyllene which are known to be attractive to E. formosa. The cuticular extracts of the nymphs predominantly contained alkanes, alkenes, and esters. Among the alkanes, synthetic nonacosane arrested the parasitoid. Our findings are discussed in relation to how the parasitoid E. formosa uses these chemicals to locate its host, T. vaporariorum.

The Role of Trialeurodes vaporariorum-Infested Tomato Plant Volatiles in the Attraction of Encarsia formosa (Hymenoptera: Aphelinidae).

Natural enemies locate their herbivorous host and prey through kairomones emitted by host plants and herbivores. These kairomones could be exploited to attract and retain natural enemies in crop fields for insect pest control. The parasitoid Encarsia formosa preferentially parasitises its whitefly host, Trialeurodes vaporariorum, a major pest of tomato Solanum lycopersicum, thus offering an effective way to improve whitefly control. However, little is known about the chemical interactions that occur in E. formosa-T. vaporariorum-S. lycopersicum tritrophic system. Using behavioural assays and chemical analyses, we investigated the kairomones mediating attraction of the parasitoid to host-infested tomato plants. In Y-tube olfactometer bioassays, unlike volatiles of healthy tomato plants, those of T. vaporariorum-infested tomato plants attracted E. formosa, and this response varied with host infestation density. Coupled gas chromatography/mass spectrometric analyses revealed that host infestation densities induced varying qualitative and quantitative differences in volatile compositions between healthy and T. vaporariorum adult-infested tomato plants. Bioassays using synthetic chemicals revealed the attractiveness of 3-carene, ß-ocimene, ß-myrcene and α-phellandrene to the parasitoid, and the blend of the four compounds elicited the greatest attraction. Our results suggest that these terpenes could be used as an attractant lure to recruit the parasitoid E. formosa for the control of whiteflies in tomato crop fields.

Re-Analysis of Abdominal Gland Volatilome Secretions of the African Weaver Ant, Oecophylla longinoda (Hymenoptera: Formicidae).

The African weaver ant, Oecophylla longinoda, is used as a biological control agent for the management of pests. The ant has several exocrine glands in the abdomen, including Dufour's, poison, rectal, and sternal glands, which are associated with pheromone secretions for intra-specific communication. Previous studies have analyzed the gland secretions of Dufour's and poison glands. The chemistry of the rectal and sternal glands is unknown. We re-analyzed the secretions from Dufour's and poison glands plus the rectal and sternal glands to compare their chemistries and identify additional components. We used the solid-phase microextraction (SPME) technique to collect gland headspace volatiles and solvent extraction for the secretions. Coupled gas chromatography-mass spectrometry (GC-MS) analysis detected a total of 78 components, of which 62 were being reported for the first time. These additional components included 32 hydrocarbons, 12 carboxylic acids, 5 aldehydes, 3 alcohols, 2 ketones, 4 terpenes, 3 sterols, and 1 benzenoid. The chemistry of Dufour's and poison glands showed a strong overlap and was distinct from that of the rectal and sternal glands. The different gland mixtures may contribute to the different physiological and behavioral functions in this ant species.

Lemon Terpenes Influence Behavior of the African Citrus Triozid Trioza erytreae (Hemiptera: Triozidae).

The African citrus triozid, Trioza erytreae Del Guercio (Hemiptera: Triozidae) is one of the primary vectors of the bacterium Candidatus Liberibacter spp. which causes citrus greening, a disease of global economic importance in citrus production. Despite its economic importance, little is known about its chemical ecology. Here, we used behavioral assays and chemical analysis to study the chemical basis of interaction between T. erytreae and one of its preferred host plants, Citrus jambhiri. In dual choice Y-tube olfactometer assays, lemon leaf odors attracted females but not males compared to plain air or solvent controls. However, in a petri dish arena assay, both sexes were arrested by lemon leaf odors. Coupled gas chromatography-mass spectrometry (GC/MS) analysis revealed quantitative differences in the odors of flushing and mature leaves, dominated by terpenes. Twenty-six terpenes were identified and quantified. In Petri dish arena assays, synthetic blends of the most abundant terpenes mimicking lemon flushing leaf odors elicited varying behavioral responses from both sexes of T. erytreae. A nine-component blend and a blend of the three most abundant terpenes; limonene, sabinene and ß-ocimene arrested both sexes of T. erytreae. In contrast, a six-component blend lacking in these three components elicited an avoidance response in both sexes. Furthermore, both sexes of T. erytreae preferred the three-component synthetic blend to lemon crude volatile extract. These results suggest that lemon terpenes might be used in the management of T. erytreae.

Repellency of Wild Oregano Plant Volatiles, Plectranthus Amboinicus, and Their Essential Oils to the Silverleaf Whitefly, Bemisia Tabaci, on Tomato.

The Bemisia tabaci (Gennadius) whitefly is a major economically damaging pest of many crops such as tomato (Solanum lycopersicum L.). Pesticides are widely used to control B. tabaci while the use of aromatic plants is an alternative control method. The aim of this study was to assess the B.tabaci repellent effect of wild oregano, Plectranthus amboinicus (Lour.) Spreng, a widespread aromatic plant in the West Indies. We tested three origins of wild oregano, including northern, central, and southern Martinique (French West Indies). Our results showed that all essential oils of wild oregano had either masking properties or were true repellents-the mean percentage of whiteflies present in the upper part of the still-air olfactometer was 1.3- to 1.9-fold lower than in the controls. The ethanolic solution of volatile organic compounds of wild oregano from southern Martinique also had a true repellent effect-the mean percentage of whiteflies present in the upper part of the still-air olfactometer was 1.3-fold lower than in the controls. Moreover, in a greenhouse insect-proof cage, there were 1.5 fewer adult whiteflies on tomato intercropped with wild oregano from southern Martinique than on tomato alone after 96 h exposure. Our study generated further insight into the potential of P. amboinicus for B. tabaci biocontrol on tomato crops. Wild oregano extracts were repellent to B. tabaci and could be used as a companion plant to prevent whitefly infestations on tomato crops. However, the B. tabaci behavior depends on the plant origin.

Repellency Potential of Tomato Herbivore-Induced Volatiles Against the Greenhouse Whitefly (Trialeurodes vaporariorum) (Hemiptera: Aleyrodidae).

The greenhouse whitefly, Trialeurode vaporariorum, is among the key pests of tomato (Solanum lycopersicum) in sub-Saharan Africa with Tuta absoluta, spider mite, thrips, and fruitworms. To understand the interaction between the pest and the plant's herbivory-induced plant volatile (HIPVs), we investigated the repellency of four tomato cultivars (Kilele F1, Assila F1, Red Beauty F1, and Nemonneta F1) upon infestation by Trialeurode vaporariorum. We analyzed the behavioral response of T. vaporariorum to infested and uninfested tomato plants of these cultivars using olfactory bioassays followed by gas chromatography-mass spectrometry (GC-MS) analyses of emitted volatiles. Trialeurode vaporariorum was attracted to uninfested plants of all four tomato cultivars. However, two cultivars Kilele F1 and Red Beauty F1 were no longer attractive to the whitefly when they were already infested by the pest. GC-MS analyses identified 25 compounds, 18 monoterpenes, 3 sesquiterpenes, 2 xylenes, 1 aldehyde, and 1 carboxylic compound in the 4 uninfested and infested cultivars. Based on the insects' behavioral response, 1,8-cineole, p-cymene, and limonene did not attract T. vaporariorum at varying concentrations when combined with Red Beauty F1, the most attractive tomato cultivar. This repellence behavioral response can be used as a basis for improvement of other vegetable crops for the management of arthropod pests as for odor masking technique.

Terpenes from herbivore-induced tomato plant volatiles attract Nesidiocoris tenuis (Hemiptera: Miridae), a predator of major tomato pests.

BACKGROUND: Biological control plays a key role in reducing crop damage by Tuta absoluta (Meyrick) and Trialeurodes vaporariorum (Westwood), which cause huge yield losses in tomato (Solanum lycopersicum L.). The mirid predator Nesidiocoris tenuis (Reuter) preys heavily on these pests, with satisfying control levels in tomato greenhouses. Although N. tenuis is known to be attracted to volatiles of tomato plants infested by T. absoluta and whitefly, little is known about the specific attractive compounds and the effect of prey density on the predator response. RESULTS: Y-tube olfactometer bioassays revealed that the attraction of N. tenuis to tomato volatiles was positively correlated with the density of T. absoluta infestation, unlike T. vaporariorum infestation. The predator was also attracted to volatiles of T. absoluta larval frass, but not to T. vaporariorum honeydew or T. absoluta sex pheromone. Among the herbivore-induced plant volatiles (HIPVs) that characterised the attractive plants infested with 20 T. absoluta larvae, olfactometer bioassays revealed that N. tenuis is attracted to the monoterpenes α-pinene, α-phellandrene, 3-carene, ß-phellandrene and ß-ocimene, whereas (E)-ß-caryophyllene was found to repel the predator. In dose-response bioassays, the five-component blend of the attractants elicited a relatively low attraction in the predator, and removal of ß-phellandrene from the blend enhanced the attraction of the predator to the resulting four-component blend, suggesting synergism among four monoterpenes. CONCLUSION: These findings suggest that a four-component blend of α-pinene, α-phellandrene, 3-carene and ß-ocimene could be used as a kairomone-based lure to recruit the predator for the biological control of T. absoluta and T. vaporariorum.

Insecticide resistance modifies mosquito response to DEET and natural repellents.

BACKGROUND: Pyrethroid and organophosphate resistance in the malaria vector Anopheles gambiae has led to the search for not only alternative insecticides, but also repellent chemical compounds. However, little is known about the potential actions of repellents and the cross-resistance risk between insecticide and repellent compounds. METHODS: Here we show the action of permethrin, DEET, geraniol, carvacrol, culminaldehyde and cinnamaldehyde against three A. gambiae strains: 'Kis' (Kisumu susceptible strain), 'KdrKis' (pyrethroid resistant strain) and 'AcerKis' (organophosphate resistant strain), the last two differing from the first by a mutation on the kdr and ace1 genes, respectively. CONCLUSIONS: Results from the DEET assays show it induced repellency for the resistant KdrKis and AcerKis strains but maintained irritancy for the susceptible strain. More generally, we show resistance genes modify the behavior of An. gambiae, increasing or decreasing the effectiveness of DEET and natural compounds, depending on the mutation. These findings offer a new avenue for research on the target and mechanism of repellent compounds. We discuss these findings in the context of vector control strategies.

A new mite IPM strategy: predator avoidance behaviour resulting from the synergetic effects of predator release and acaricide-treated nets.

BACKGROUND: Tetranychus evansi and T. urticae spider mites are known major pests of Solanaceae. Smallholders in Africa rely heavily on pesticide treatments. However, farmers claim that pesticides are generally ineffective despite high-frequency sprays. New management solutions are thus urgently needed. This study assessed the efficacy of using acaricide-treated nets combined with predatory mite release for controlling spider mites. RESULTS: The results showed the acaricide-treated net alone was more effective at reducing numbers of T. urticae than T. evansi. We observed the opposite for release of the predatory mite Phytoseiulus longipes. This difference could be explained by the specific dispersion strategies of the two spider mite pests; T. evansi is gregarious, whereas T. urticae dispersed rapidly. Joint application of both techniques resulted in a synergetic effect that reduced T. evansi and T. urticae spider mite numbers close to zero. The synergetic effect could be explained by predator avoidance behaviour of the prey spider mites, resulting in higher prey trapping and killing rates on acaricide-treated nets, while P. longipes fed on spider mite eggs. CONCLUSION: These techniques are profitable for smallholders as they are not expensive and avoid residues on the crop. © 2018 Society of Chemical Industry.

Performance of Metarhizium anisopliae-treated foam in combination with Phytoseiulus longipes Evans against Tetranychus evansi Baker & Pritchard (Acari: Tetranychidae).

BACKGROUND: Tetranychus evansi (Te) is an exotic pest of solanaceous crops in Africa. The predatory mite Phytoseiulus longipes (Pl) and the fungus Metarhizium anisopliae (Ma) are potential biocontrol agents of Te. The present study investigated the efficacy of fungus-treated foam placed above or below the third Te-infested tomato leaf. The persistence of fungus-treated foam and the performance of Pl with and without fungus-treated foam were evaluated. RESULTS: The fungus-treated foam was effective when Te infestation was below the third tomato leaf as no damage was recorded on any of the upper tomato leaves up to 30 days post-treatment. However, in the control treatments, the infestation increased considerably from 9 ± 0.3% to 100 ± 0% (mean ± standard error) at 15 days post-treatment. The reuse of the fungus-treated foam at 15, 30 and 45 days post-treatment resulted in 19 ± 1.4%, 25 ± 1.2% and 54 ± 2.1%, respectively, infestation by Te. The fungus-treated foam and Pl alone were efficient, but there was no benefit to combining them for use against Te. CONCLUSION: The fungus-treated foam is an effective method to optimise the use of Ma in screenhouse conditions. These two control agents could be integrated in an integrated pest management strategy for crop protection. However, these results need to be confirmed in large field trials. © 2018 Society of Chemical Industry.

Naturally occurring bioactive compounds from four repellent essential oils against Bemisia tabaci whiteflies.

BACKGROUND: In tropical countries, netting is an effective sustainable tool for protecting horticultural crops against Lepidoptera, although not against small pests such as Bemisia tabaci, while smaller mesh netting can be used in temperate regions. A solution is to combine a net with a repellent. Previously we identified repellent essential oils: lemongrass (Cymbopogon citratus), cinnamon (Cinnamomum zeylanicum), cumin (Cuminum cyminum) and citronella (Cymbopogon winternarius). The present study was designed to identify the active compounds of these essential oils, characterise their biological activity and examine their potential for coating nets. We investigated the efficiency and toxicity of nets dipped in different solutions. We then studied the repellent effect with an olfactometer and the irritant effect by videotracking. RESULTS: Geraniol and citronellol were the most promising net coatings owing to their repellent effect. The repellency, irritancy or toxicity varied with the product and concentration, and these features were independent, indicating that the repellent and the irritant/toxic mechanisms were not the same. The combined effects of these different compounds account for the bioactivity of the mixture, suggesting interactions between the compounds. CONCLUSION: This new sustainable strategy for protecting vegetable crops against whiteflies is discussed, in addition to the use of companion plants that could produce such bioactive compounds.

Electrophysiological and behavioral characterization of bioactive compounds of the Thymus vulgaris, Cymbopogon winterianus, Cuminum cyminum and Cinnamomum zeylanicum essential oils against Anopheles gambiae and prospects for their use as bednet treatments.

BACKGROUND: Laboratory and field studies showed that repellent, irritant and toxic actions of common public health insecticides reduce human-vector contact and thereby interrupt disease transmission. One of the more effective strategies to reduce disease risk involves the use of long-lasting treated bednets. However, development of insecticide resistance in mosquito populations makes it imperative to find alternatives to these insecticides. Our previous study identified four essential oils as alternatives to pyrethroids: Thymus vulgaris, Cymbopogon winterianus, Cuminum cyminum, Cinnamomum zeylanicum. The objectives of this study were to identify active compounds of these essential oils, to characterize their biological activity, and to examine their potential as a treatment for bednets. METHODS: We evaluated the electrophysiological, behavioural (repellency, irritancy) and toxic effects of the major compounds of these oils against Anopheles gambiae strain 'Kisumu'. RESULTS: Aldehydes elicited the strongest responses and monoterpenes the weakest responses in electroantennogram (EAG) trials. However, EAG responses did not correlate consistently with results of behavioral assays. In behavioral and toxicity studies, several of the single compounds did exhibit repellency, irritancy or toxicity in An. gambiae; however, the activity of essential oils did not always correlate with activity expected from the major components. On the contrary, the biological activity of essential oils appeared complex, suggesting interactions between individual compounds and the insect under study. Data also indicated that the three effects appeared independent, suggesting that repellency mechanism(s) may differ from mechanisms of irritancy and toxicity. CONCLUSIONS: Based on the bioassays reported here, some of the compounds merit consideration as alternative bednet treatments.

Repellent, irritant and toxic effects of 20 plant extracts on adults of the malaria vector Anopheles gambiae mosquito.

Pyrethroid insecticides induce an excito-repellent effect that reduces contact between humans and mosquitoes. Insecticide use is expected to lower the risk of pathogen transmission, particularly when impregnated on long-lasting treated bednets. When applied at low doses, pyrethroids have a toxic effect, however the development of pyrethroid resistance in several mosquito species may jeopardize these beneficial effects. The need to find additional compounds, either to kill disease-carrying mosquitoes or to prevent mosquito contact with humans, therefore arises. In laboratory conditions, the effects (i.e., repellent, irritant and toxic) of 20 plant extracts, mainly essential oils, were assessed on adults of Anopheles gambiae, a primary vector of malaria. Their effects were compared to those of DEET and permethrin, used as positive controls. Most plant extracts had irritant, repellent and/or toxic effects on An. gambiae adults. The most promising extracts, i.e. those combining the three types of effects, were from Cymbopogon winterianus, Cinnamomum zeylanicum and Thymus vulgaris. The irritant, repellent and toxic effects occurred apparently independently of each other, and the behavioural response of adult An. gambiae was significantly influenced by the concentration of the plant extracts. Mechanisms underlying repellency might, therefore, differ from those underlying irritancy and toxicity. The utility of the efficient plant extracts for vector control as an alternative to pyrethroids may thus be envisaged.

The repellency of lemongrass oil against stable flies, tested using video tracking.

Lemongrass oil (Cymbopogon citratus) is an effective repellent against mosquitoes (Diptera: Culicidae) and house flies (Diptera: Muscidae). In this study, its effectiveness was assessed on stable flies (Diptera: Muscidae) in laboratory conditions. First, we demonstrated that lemongrass oil is an active substance for antennal olfactory receptor cells of Stomoxys calcitrans as indicated by a significant increase in the electroantennogram responses to increasing doses of lemongrass oil. Feeding-choice tests in a flight cage with stable flies having access to two blood-soaked sanitary pads, one of which was treated with lemongrass oil, showed that stable flies (n = 24) spent significantly more time in the untreated zone (median value = 218.4 s) than in the treated zone (median value = 63.7 s). No stable flies fed on the treated pad, whereas nine fed on the untreated pad. These results suggest that lemongrass oil could be used as an effective repellent against stable flies. Additional studies to confirm its spatial repellent and feeding deterrent effects are warranted.