Sequential Isolation of Essential Oils Repellent to the Red Palm Weevil Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae).

The push-pull approach using semiochemicals in pest control requires both an attractant and a repellent. Many previous studies have arbitrarily tested one or more known insect repellents or plant essential oils (EOs) hoping to find repellents of an insect pest. We used a comprehensive approach that synergistically tests in the field numerous natural volatiles from commercial EOs to identify repellents of the red palm weevil Rhynchophorus ferrugineus (Coleoptera: Curculionidae), a worldwide pest of palms and date palms. Volatiles from 79 EOs in slow-release devices were divided into five groups and tested in traps with attractive pheromone compared to traps with pheromone alone. EO-treatment groups exhibiting repellency due to significant trap shutdown, were further subdivided into subgroups of four EOs each and tested further. Two groups of four EOs (cypress, desert wormwood, elemi, and Eucalyptus citriodora) and (niaouli, nutmeg, oregano, and orange sweet), or their corresponding mixtures of major volatiles, caused pheromone trap reductions of up to 92%. Further tests showed that seven of the eight EOs are similarly repellent as the corresponding subgroup. This systematic approach of successively testing sub-fractions of EOs in the field for trap shutdown should be useful to identify repellents of other insect pests of crops.

Modelling push-pull management of pest insects using repellents and attractive traps in fruit tree orchards.

BACKGROUND: Push-pull with semiochemicals in pest management uses repellents to reduce response of pests to food-mate resources (push) and attractive traps to reduce populations (pull). Simulation models of push-pull can aid understanding of plant-insect interactions in nature and suggest hypotheses for field tests that improve management. A previous model indicated advantages of push-pull for controlling ambrosia beetle, Euwallacea fornicatus, pest of avocado trees. However, the simulated behavior of repellency was inconsistent with field observations. RESULTS: We simulated individual-based movement of insects in push-pull to reveal relationships between parameters of radii (strength) of attractive traps, pest aggregations, and repellents with densities of each in an avocado orchard to visualize and understand the interactions and significance. Simulations indicated placement of traps along a 1-ha area periphery as a barrier resulted in similar trapping and mating as when traps were in a grid, either when insects originated randomly inside the plot or came from outside the plot. However, when insects originated from outside, both arrangements caught slightly more than when insects originated within the plot. CONCLUSION: There were no differences in capture rates whether traps were spaced in a barrier along the plot's periphery or in a grid covering the plot. Push-pull was more effective than pull (mass trapping) alone. Repellent behavior of individuals when encountering a repellent radius was modelled by approximate 90° turns (right or left at random) when about to enter an infestation, consistent with earlier observations of effects of repellents on bark beetles orienting to aggregation pheromone. © 2022 Society of Chemical Industry.

Circadian rhythms of insect pheromone titer, calling, emission, and response: a review.

Many insect species have circadian rhythms of pheromone production/titer, calling, emission, and response that are involved in intraspecific communication and impact pest management practices. Rhythms of pheromone biosynthesis, most studied in moths affecting forestry and agriculture, contribute to a periodicity of pheromone concentration or titer within glands or hemolymph. Calling rhythms by the pheromone-emitting sex are physical movements (pumping, vibrating wings) that aid in release and dispersion of the volatile pheromone components attractive to the opposite conspecific sex or both sexes. Circadian rhythms of emission of pheromone also occur as a result of an interaction between calling and the titer of pheromone available for release. Responding individuals usually show a coincidental rhythm of dispersal flight while seeking pheromone plumes in which, by orienting upwind, the insects find mates or food resources. However, some species begin searching an hour or more before the emitting sex initiates calling and emission, which benefits mass trapping control programs because the baited traps do not compete initially with natural pheromone sources. In our review, data of daily rhythms of moths and other insects were extracted from the literature by screen capture software to calculate mean time of activity and standard deviation and fit to normal curves. These methods are illustrated for various insects and as a basis for discussion of interactions of pheromonal circadian rhythms of the well-studied gypsy moth Lymantria dispar, spruce budworm moth Choristoneura fumiferana, turnip moth Agrotis segetum, and cabbage looper moth Trichoplusia ni. The various circadian rhythms are discussed in relation to application of species-specific sex and aggregation pheromones for benign biological control and management of pest insects.

Absolute Configuration of the Spherical Mealybug Nipaecoccus viridis Sex Pheromone, γ-Necrodyl Isobutyrate: Chemoenzymatic Synthesis and Bioassays.

The spherical mealybug, Nipaecoccus viridis (Hemiptera: Pseudococcidae), is a major global pest causing feeding damage to leaves and fruits of citrus varieties, soybean, mango, pomegranate, and grapevines. Females of the mealybug release a sex pheromone that was identified recently as a mixture of γ-necrodyl isobutyrate and γ-necrodol. The identification required synthesis based on a natural source of trans-α-necrodol, of unknown chirality, obtained from essential oil of Spanish lavender, Lavandula luisieri. To determine the chirality of the sex pheromone, here, we synthesize the γ-necrodyl acetate enriched in (+)-(S)-enantiomer and separate the enantiomers using a lipase enzyme. We confirm that the natural components, both in the mealybug and in the lavender essential oil, consist of (-)-(R)-enantiomers. Bioassays conducted in the lab and field show that males are attracted to (-)-(R)-γ-necrodyl isobutyrate alone.

Semiochemicals Affecting Attraction of Ambrosia Beetle Euwallacea fornicatus (Coleoptera: Curculionidae: Scolytinae) to Quercivorol: Developing Push-Pull Control.

Euwallacea fornicatus (Eichhoff), the polyphagous shot hole borer (PSHB), is an ambrosia beetle infesting avocado Persea americana Mill. limbs in North America and Israel. We conducted field experiments with sticky traps in avocado orchards to develop push-pull semiochemical methods of managing PSHB. Traps baited with 10-fold increasing doses (0.01 to 100× or 1.26 µg to 12.6 mg/d) of attractant quercivorol were previously shown to increasingly capture female PSHB (males flightless). We converted trap catch of this relationship to a standardized effective attraction radius (EAR) that predicts capture power of baited-traps regardless of insect flight density. Earlier, piperitone and verbenone were shown to strongly inhibit attraction of PSHB to quercivorol-traps. We tested increasing numbers of 1× piperitone dispensers at 0.75-m distance surrounding a quercivorol-trap and found PSHB catch to decline exponentially. Increasing decadic doses (0.01 to 10×) of either verbenone or piperitone released at 1× quercivorol-traps caused a sigmoidal first-order kinetic-decay in catch. Verbenone (1×) placed at increasing distances (0, 0.25, 0.5, 1, and 2 m) from a 1× quercivorol-trap became increasingly ineffective in reducing catch of PSHB. We found no evidence that ethanol released from 7.5 to 480 mg/d affected attraction of PSHB, but Scobicia chevrieri (Villa and Villa) (Coleoptera: Bostrichidae) was increasingly attracted. Due to their relatively short-range (<0.5 m) inhibition of attractive sources, piperitone, and verbenone dispensers should be placed on avocado trunks where PSHB aggregations occur before the flight season.

Magnetic Polyurea Nano-Capsules Synthesized via Interfacial Polymerization in Inverse Nano-Emulsion.

Polyurea (PU) nano-capsules have received voluminous interest in various fields due to their biocompatibility, high mechanical properties, and surface functionality. By incorporating magnetic nanoparticle (MNPs) into the polyurea system, the attributes of both PU and MNPs can be combined. In this work, we describe a facile and quick method for preparing magnetic polyurea nano-capsules. Encapsulation of ionic liquid-modified magnetite nanoparticles (MNPs), with polyurea nano-capsules (PU NCs) having an average size of 5-20 nm was carried out through interfacial polycondensation between amine and isocyanate monomers in inverse nano-emulsion (water-in-oil). The desired magnetic PU NCs were obtained utilizing toluene and triple-distilled water as continuous and dispersed phases respectively, polymeric non-ionic surfactant cetyl polyethyleneglycol/polypropyleneglycol-10/1 dimethicone (ABIL EM 90), diethylenetriamine, ethylenediamine diphenylmethane-4,4'-diisocyanate, and various percentages of the ionic liquid-modified MNPs. High loading of the ionic liquid-modified MNPs up to 11 wt% with respect to the dispersed aqueous phase was encapsulated. The magnetic PU NCs were probed using various analytical instruments including electron microscopy, infrared spectroscopy, X-ray diffraction, and nuclear magnetic spectroscopy. This unequivocally manifested the successful synthesis of core-shell polyurea nano-capsules even without utilizing osmotic pressure agents, and confirmed the presence of high loading of MNPs in the core.

Identification of the Sex Pheromone of the Spherical Mealybug Nipaecoccus viridis.

The spherical mealybug Nipaecoccus viridis is a pest of several major crops including soybeans, grapes and citrus varieties. Sessile virgin females of N. viridis release two volatiles, 2,2,3,4-tetramethyl-3-cyclopentene-1-methanol (γ-necrodol) and γ-necrodyl isobutyrate, on a circadian rhythm with peak at 17:00 (11 hr of photophase) as determined by automated, sequential solid phase micro extraction with gas chromatography-mass spectrometry analysis. The females increased the released amounts with age by about seven-fold from 5 to 6 d to 10-12 d of age. trans-3,4,5,5-Tetramethyl-2-cyclopentene-1-methanol (trans-α-necrodol) and trans-α-necrodyl acetate, found in essential oil of Spanish lavender, Lavandula luisieri, were rearranged to γ-necrodol and then used to synthesize γ-necrodyl isobutyrate. GC-MS and NMR data confirmed the identifications. In a petri dish bioassay, N. viridis males were significantly attracted to filter paper discs impregnated with γ-necrodyl isobutyrate but not to γ-necrodol or controls. A mixture of the two compounds was not more attractive than γ-necrodyl isobutyrate alone. Similar results were obtained with trapping flying adults, suggesting that the sex pheromone consists only of γ-necrodyl isobutyrate. This compound has not been reported previously in insects. Conversion of α-necrodol in lavender essential oil simplifies the synthesis of the sex pheromone and should allow its use in management of this cosmopolitan invasive pest.

Index of host habitat preference explored by movement-based simulations and trap captures.

Animal species likely have different strengths of host habitat preference (HHP) that might be characterized by a standardized index ranging from 0 (no preference) to 1 (maximum preference). We hypothesized that in some species, HHP may result from individuals dispersing out of the host habitat having a probability of turning back at the boundary, or after entering host habitat by reducing speed or increasing size of turning angles. Computer simulations of individuals moving between various sized patches of host and nonhost habitat were conducted based on these three behaviours hypothesized to affect HHP. In the rebounding model, simulations resulted in equilibria of animal numbers inside and outside of host habitat that depend on sizes of these areas, initial number and the rebounding probability. Curvilinear regression of simulation results suggested an equation that predicted numbers in the host habitat and was solved for rebounding probability. A modified equation that sampled population densities (e.g., insect pheromone trap catches) inside and outside host habitat areas gave the rebounding probability, an index of HHP, without requiring the sizes of the areas. Simulations with traps and moving animals verified that the modified equation could predict the index correctly. The modified equation also estimates an index of HHP from sampled densities due to speed reductions and a combination of this and rebounding. Changes in angular turning size upon entering host habitat, however, did not affect habitat preference. Using pheromone trap captures, we found that the lesser date moth Batrachedra amydraula has a HHP for date Phoenix dactylifera plantations of 0.96. Host habitat preference indexes also were calculated from sampled insect densities reported in the literature. The new index of HHP is useful to characterize habitat patches of many organisms and aid understanding of animal spatial distributions and speciation processes. In addition, the index can be applied in studies of invasive species, trap crops of pest insects and conservation management.

Inhibitory Effects of Semiochemicals on the Attraction of an Ambrosia Beetle Euwallacea nr. fornicatus to Quercivorol.

The Euwallacea fornicatus (Eichhoff) species complex includes the polyphagous shot hole borer (PSHB), an ambrosia beetle infesting avocado limbs, Persea americana Mill. Synthetic quercivorol, a monoterpene alcohol, is known to attract females (males are flightless) over a range of release rates spanning three orders of magnitude. The upper release dose was extended 10-fold using sticky traps baited with quercivorol released at 1× (0.126 mg/day), 10×, and 108× relative rates to obtain a dose-response curve fitting a kinetic formation function. Naturally infested limbs of living avocado trees were wrapped with netting to exclude the possibility of catching emerging beetles on the encircling sticky traps. The results indicate PSHB are significantly attracted to infested limbs. Ethanol released over a 64-fold range (lowest rate of 7.5 mg/day) was moderately inhibitory of PSHB attraction to 1× quercivorol. ß-caryophyllene and eucalyptol did not appear to affect attraction at the rates tested. A field test of potential inhibitors of 1× quercivorol was done using ~1 mg/day releases of monoterpene ketones: (-)-(S)-verbenone, (+)-(R)-verbenone, 3-methyl-2-cyclo-hexen-1-one (MCH or seudenone), piperitone, (+)-(S)-carvone, and racemic cryptone. Only piperitone and the two enantiomers of verbenone were strongly inhibitory. A blend of piperitone and verbenone tested together at different distances (0, 0.5, 1, 2, and 4 m) from a 1× quercivorol baited sticky trap became increasingly ineffective in inhibiting the attractant as separation distance increased. Due to the relatively short-range repellency (<1 m), the inhibitors would need to be released from several places on each tree to effectively repel PSHB from avocado trees. Effective attraction radii, EAR, and circular EARc are estimated for the quercivorol baits released at 1×, 10× and 108× rates. Push-pull simulations of moving beetles were performed in 1 ha plots with 2, 4, or 16 traps of 10× EARc and 400 trees (0, 1, or 3 inhibitors per tree) of which ten had an infested limb (EARc = 0.5 m). The simulations indicate that push-pull methods would be more effective in reducing PSHB mating than simply using mass-trapping alone.

An insect anti-antiaphrodisiac.

Passive mechanisms of mate guarding are used by males to promote sperm precedence with little cost, but these tactics can be disadvantageous for their mates and other males. Mated females of the plant bug Lygus hesperus are rendered temporarily unattractive by seminal fluids containing myristyl acetate and geranylgeranyl acetate. These antiaphrodisiac pheromones are gradually released from the female's gonopore, declining until they no longer suppress male courtship. Because starting quantities of these compounds can vary widely, the repellant signal becomes less reliable over time. Evidence was found of a complimentary mechanism that more accurately conveys female mating status. Once inside the female, geranylgeranyl acetate is progressively converted to geranylgeraniol then externalized. Geranylgeraniol counteracts the antiaphrodisiac effect despite having no inherent attractant properties of its own. This is the first evidence for such an anti-antiaphrodisiac pheromone, adding a new element to the communication mechanisms regulating reproductive behaviors.

Attraction of the Euwallacea sp. near fornicatus (Coleoptera: Curculionidae) to Quercivorol and to Infestations in Avocado.

The Euwallacea sp. near fornicatus (Euwallacea sp. 1 hereafter) feeds on many woody shrubs and trees and is a pest of avocado, Persea americana Mill., in several countries including Israel and the United States. Quercivorol baits are commercially available for Euwallacea sp. 1 females (males do not fly), but their attractive strength compared to other pheromones and potential for mass trapping are unknown. We used sticky traps baited with quercivorol released at 0.126 mg/d (1×) and at 0.01×, 0.1×, and 10× relative rates to obtain a dose-response curve of Euwallacea sp. 1 attraction. The curve fitted well a kinetic formation function of first order. Naturally infested limbs of living avocado trees had attraction rates equivalent to 1× quercivorol. An effective attraction radius (EAR) was calculated according to previous equations for each of the various baits (1× EAR = 1.18 m; 10× EAR = 2.00 m). A pole with six sticky traps spaced from 0.25-5.75 m in height had captures of Euwallacea sp. 1 yielding a mean flight height of 1.24 m with vertical flight distribution SD of 0.88 m (0.82-0.96 m, 95% CI). The SD with specific EAR was used to calculate EARc, two-dimensional EAR (1× EARc = 0.99 m; 10× EARc = 2.86 m), for comparison with other insect pheromone traps and for use in simulations. The simulation methods described previously were performed with combinations of 1-16 traps with 1-50 aggregations per 9-ha plot. The simulations indicate mass trapping with quercivorol could be effective if begun in spring before Euwallacea sp. 1 establishes competing sources of attraction.

Diel periodicity of pheromone release by females of Planococcus citri and Planococcus ficus and the temporal flight activity of their conspecific males.

The diel periodicity of sex pheromone release was monitored in two mealybug species, Planococcus citri and Planococcus ficus (Hemiptera; Pseudococcidae), using sequential SPME/GCMS analysis. A maximal release of 2 ng/h pheromone by 9-12-day-old P. citri females occurred 1-2 h before the beginning of photophase. The highest release of pheromone by P. ficus females was 1-2 ng/2 h of 10-20-day-old females, approximately 2 h after the beginning of photophase. Mating resulted in termination of the pheromone release in both mealybug species. The temporal flight activity of the males was monitored in rearing chambers using pheromone baited delta traps. Males of both P. citri and P. ficus displayed the same flight pattern and began flying at 06:00 hours when the light was turned on, reaching a peak during the first and second hour of the photophase. Our results suggest that other biparental mealybug species display also diel periodicities of maximal pheromone release and response. Direct evaluation of the diel periodicity of the pheromone release by the automatic sequential analysis is convenient and will be very helpful in optimizing the airborne collection and identification of other unknown mealybug pheromones and to study the calling behavior of females. Considering this behavior pattern may help to develop more effective pheromone-based management strategies against mealybugs.

Circadian release of male-specific components of the greater date moth, Aphomia (Arenipses) sabella, using sequential SPME/GC/MS analysis.

The greater date moth (GDM), Aphomia sabella Hampson (Lepidoptera: Pyralidae: Galleriinae), is a serious pest of date palms, Phoenix dactylifera. The release of volatiles from both males and females was investigated using sequential SPME/GC/MS analysis. Males release a complex mixture of compounds in a circadian rhythm during the night between 03:00 and 05:00 hr. Six compounds were identified: benzaldehyde, sulcatol (6-methyl-5-hepten-2-ol), geranyl acetone [(E)-6,10-dimethyl-5,9-undecadien-2-one], phenylacetaldehyde, 2-phenylpropenal, and (R)-fuscumol [(R)-(E)-6, 10-dimethyl-5, 9-undecadien-2-ol]. Benzaldehyde, sulcatol, and geranyl acetone were found only in trace amounts. These compounds were in glands located in the forewing of males only. Small amounts of acetoin and 2,3-butanediol were found sporadically in the SPME/GC/MS analyses of volatiles from females, and these compounds probably originate from microorganisms. This is the first finding of a circadian release of male-specific compounds in moths. GC/EAD analyses with synthetic standards of compounds released by males showed that the female antenna is stimulated by all six compounds, while the male antenna responded only to phenylacetaldehyde. A possible pheromonal role for the male-specific compounds is suggested by the circadian rhythm of their release and the EAD response of females to them. However, trapping tests with the main male-specific compounds in screen cages in the laboratory or in the field did not reveal any significant behavioral responses from females or males. Copulation in the laboratory was observed only in the presence of date palm tissue, thus suggesting that sexual communication and mating of GDM moths probably occurs in the crown of date palms.

Sex pheromone component ratios and mating isolation among three Lygus plant bug species of North America.

The plant bugs Lygus hesperus, Lygus lineolaris, and Lygus elisus (Hemiptera: Miridae) are major pests of many agricultural crops in North America. Previous studies suggested that females release a sex pheromone attractive to males. Other studies showed that males and females contain microgram amounts of (E)-4-oxo-2-hexenal, hexyl butyrate, and (E)-2-hexenyl butyrate that are emitted as a defense against predators. Using gas chromatography-mass spectrometry, we found that female L. lineolaris and L. elisus have a 4:10 ratio of hexyl butyrate to (E)-2-hexenyl butyrate that is reversed from the 10:1 ratio in female L. hesperus (males of the three species have ~10:1 ratio). These reversed ratios among females of the species suggest a behavioral role. Because both sexes have nearly equal amounts of the major volatiles, females should release more to attract males. This expectation was supported because L. hesperus females released more hexyl butyrate (mean of 86 ng/h) during the night (1800-0700 hours) than did males (<1 ng/h). We used slow-rotating pairs of traps to test the attraction of species to blends of the volatiles with a subtractive method to detect synergism. Each species' major butyrate ester was released at 3 µg/h, the minor butyrate according to its ratio, and (E)-4-oxo-2-hexenal at 2 µg/h. The resulting catches of only Lygus males suggest that (E)-4-oxo-2-hexenal is an essential sex pheromone component for all three species, (E)-2-hexenyl butyrate is essential for L. elisus and L. lineolaris, and hexyl butyrate is essential for L. hesperus. However, all three components are recognized by each species since ratios of the butyrate esters are critical for conspecific attraction and heterospecific avoidance by males and thus play a role in reproductive isolation among the three species. Because L. hesperus males and females are known to emit these major volatiles for repelling ant predators, our study links defensive allomones in Lygus bugs with an additional use as sex pheromones.

Analyzing diurnal and age-related pheromone emission of the olive fruit fly, Bactrocera oleae by sequential SPME-GCMS analysis.

The olive fruit fly, Bactrocera oleae (Diptera: Tephritidae), uses 1,7-dioxaspiro[5.5]undecane ("olean"), produced primarily by females, as a sex pheromone. We used sequential solid phase microextraction-gas chromatography mass spectrometry (SPME-GCMS) analysis to show that female olive flies release about 1000 ng of pheromone at the onset of scotophase for several weeks, while males release about 1/100 as much during the first week after eclosion. The present research demonstrates details of employing SPME-GCMS with the partially known pheromone system of the olive fruit fly as a model for pheromone identification and diurnal release patterns in insects, especially fruit flies. The sequential SPME-GCMS method will readily allow detection and semi-quantification of semiochemicals released by insects in minute amounts throughout the diurnal cycle.

Male behaviors reveal multiple pherotypes within vine mealybug Planococcus ficus (Signoret) (Hemiptera; Pseudococcidae) populations.

The vine mealybug (VM) females collected in Israel produce two sex pheromone compounds: lavandulyl senecioate (LS) and (S)-lavandulyl isovalerate (LI). The males display ambiguous behavior to LI: repulsion in the vineyard and attraction of laboratory-reared males. We addressed the question of individual male behavior, i.e., do males respond to both LS and LI, or might they display a distinct response to each of the two pheromone compounds. We compared male pherotype frequencies between wild-caught and laboratory-reared populations. Then, we examined the relationship between pherotype composition and male capture rates in pheromone traps. Finally, we addressed the heredity of the pherotypes. The Israeli VM populations contain nine different male pherotypes, as defined according to the male behavior to pheromone compounds. The studied Portuguese populations included five of the nine pherotypes; none of the Portuguese males were attracted to LI. It seems that the high frequency of males that were attracted to LI is related to dense VM populations. It is hypothesized that selection for the male pherotypes, I males, those that respond to LI, occur under high-density rearing conditions. This may result from shorter development times of males and females that produce more I male pherotypes. The lower relative frequency of trapping of males in LI-baited traps than expected from the percentage determined in a Petri dish arena suggests that males that respond solely to LS (S males) are better fliers. The results also suggest that the pherotype trait is inherited by both sexes of the VM.