Parasitic wasps avoid ant-protected hemipteran hosts via the detection of ant cuticular hydrocarbons.

One of the most studied and best-known mutualistic relationships between insects is that between ants and phloem-feeding insects. Ants feed on honeydew excreted by phloem-feeding insects and, in exchange, attack the phloem feeders' natural enemies, including parasitic wasps. However, parasitic wasps are under selection to exploit information on hazards and avoid them. Here, we tested whether parasitic wasps detect the previous presence of ants attending colonies of phloem feeders. Behavioural assays demonstrate that wasps left colonies previously attended by ants more frequently than control colonies. This behaviour has a potential cost for the parasitic wasp as females inserted their ovipositor in fewer hosts per colony. In a further bioassay, wasps spent less time on papers impregnated with extracts of the ant cues than on control papers. Gas chromatography coupled with mass spectrometry analyses demonstrated that ants left a blend of cuticular hydrocarbons when they attended colonies of phloem feeders. These cuticular hydrocarbons are deposited passively when ants search for food. Overall, these results suggest, for the first time, that parasitic wasps of honeydew producers detect the previous presence of mutualistic ants through contact infochemicals. We anticipate such interactions to be widespread and to have implications in numerous ecosystems, as phloem feeders are usually tended by ants.

Geographic Location, Population Dynamics, and Fruit Damage of an Invasive Citrus Mealybug: The Case of Delottococcus aberiae De Lotto in Eastern Spain.

The invasive mealybug Delottococcus aberiae De Lotto (Hemiptera: Pseudococcidae) has rapidly spread in the Mediterranean basin since its detection in 2009 in the Valencia Community in eastern Spain. The use of sticky traps baited with its sex pheromone, (4,5,5-trimethyl-3-methylenecyclopent-1-en-1-yl)methyl acetate, has allowed to determine the geographical distribution of D. aberiae by means of the surveillance network described in the present work. The population monitoring of the pest over a five-year period (2019-2023) has revealed an increase from 31% to 70% of the affected citrus-growing area. The monitoring network has also allowed a better understanding of the pests' biological cycle throughout the year. The populations start growing from March to June and reach their maximum in July-August. During autumn, there is a gradual decline in the population. Although the highest annual populations were detected in 2022 and 2023, the greatest crop losses were recorded in 2020 and 2021, with mean values near 18%. Data suggest that the damage responsible for fruit deformation, and thus the economic losses, are related to the population levels in spring (April-May) rather than those in summer (July-August). The findings of this study can be valuable for future research and development of effective pest control strategies.

Effect of Commercial Trap Design and Location on Captures of Diocalandra frumenti (Fabricius) (Coleoptera: Dryophthoridae) on Palm Trees.

Diocalandra frumenti (Fabricius) (Coleoptera: Dryophthoridae) is a weevil present in the Canary Islands, affecting economically important palms such as Phoenix canariensis H. Wildpret and its hybrids, for which there were no trapping tools. The larvae cause the main damage by burrowing galleries in the rachis of the leaves, causing premature drying and collapse. To develop an effective trap, six trials were carried out to evaluate the effect of trap type, design, colour, height, distance and location of the trap in relation to the palm tree on D. frumenti captures. This study confirms that the Econex® trap, green in colour, without a cover and with two ventilation holes of 2.5 cm in diameter, diametrically opposite each other and at 1 cm from the top of the base of the trap, baited with sugar cane and water, and placed between the first and second ring of green leaves of the palm canopy, is efficient in capturing D. frumenti. These results establish a basis for future research focused on the development of a specific trapping system based on semiochemicals to serve as a tool for detection, monitoring and mass trapping of D. frumenti.

Absolute Configuration of the Invasive Mealybug Delottococcus aberiae (De Lotto) Sex Pheromone: Synthesis and Bioassay of Both Enantiomers.

The mealybug Delottococcus aberiae (De Lotto) (Hemiptera: Pseudococcidae) is an invasive pest reported in Europe at the end of the first decade of the 2000s, causing severe damage to citrus production in eastern Spain. In a previous work, (4,5,5-trimethyl-3-methylenecyclopent-1-en-1-yl)methyl acetate was identified as the sex pheromone emitted by females, a new compound with an unusual ß-necrodol skeleton possessing one stereocenter. This compound was assigned to the (-)-enantiomer but the absolute configuration was then not reported. In the present study, enantiomeric pure samples of both enantiomers were synthesized. X-ray diffraction analysis allowed the (-)-enantiomer, identical to the one emitted by virgin D. aberiae females, to be unequivocally identified as (-)-(R)-(4,5,5-trimethyl-3-methylenecyclopent-1-en-1-yl)methyl acetate. Bioassays carried out to test the activity of both enantiomers under field conditions suggest that the presence of the (+)-(S)-enantiomer has detrimental effects on the activity of the racemates.

New Monoterpenoid as the Sex Pheromone of Spanish Populations of the Longtailed Mealybug Pseudococcus Longispinus (Hemiptera: Pseudococcidae).

Pseudococcus longispinus (Targioni-Tozzetti) (Hemiptera: Coccoidea: Pseudococcidae), a polyphagous and cosmopolitan pest native to Australia, is a highly damaging pest for numerous crops of economic importance. The sex pheromone of this species (2-(1,5,5-trimethylcyclopent-2-en-1-yl)ethyl acetate), currently used for pest monitoring purposes, was not attractive to males in field experiments conducted in Spanish persimmon orchards infested with this mealybug. The virgin and mated female volatile profiles of these P. longispinus populations were studied by the volatile collection of effluvia in Porapak-Q. The resulting extracts were analyzed by gas chromatography coupled to mass spectrometry (GC-MS), revealing a new compound specific to virgin females and different from the previously described sex pheromone. Based on GC-MS data and nuclear magnetic resonance experiments, we envisaged monoterpene 2-(1,5-dimethyl-4-methylenecyclopent-2-en-1-yl)ethyl acetate as the new sex pheromone candidate, which was synthesized and shown to be attractive in the field to P. longispinus males of the Spanish population.

Transcriptome-informed identification and characterization of Planococcus citri cis- and trans-isoprenyl diphosphate synthase genes.

Insect physiology and reproduction depend on several terpenoid compounds, whose biosynthesis is mainly unknown. One enigmatic group of insect monoterpenoids are mealybug sex pheromones, presumably resulting from the irregular coupling activity of unidentified isoprenyl diphosphate synthases (IDSs). Here, we performed a comprehensive search for IDS coding sequences of the pest mealybug Planococcus citri. We queried the available genomic and newly generated short- and long-read P. citri transcriptomic data and identified 18 putative IDS genes, whose phylogenetic analysis indicates several gene family expansion events. In vitro testing confirmed regular short-chain coupling activity with five gene products. With the candidate with highest IDS activity, we also detected low amounts of irregular coupling products, and determined amino acid residues important for chain-length preference and irregular coupling activity. This work therefore provides an important foundation for deciphering terpenoid biosynthesis in mealybugs, including the sex pheromone biosynthesis in P. citri.

The Dual Benefit of Plant Essential Oils against Tuta absoluta.

Plant essential oils (PEOs) are being studied as a potential alternative to synthetic pesticides in agriculture. PEOs have the potential to control pests both directly, by being toxic or repellent to pests, and indirectly, by activating plant's defense mechanisms. In this study, the effectiveness of five PEOs (Achillea millefolium, Allium sativum, Rosmarinus officinallis, Tagetes minuta, and Thymus zygis) on controlling Tuta absoluta and their impact on the predator Nesidiocoris tenuis was examined. The study revelead that PEOs from A. millefolium and A. sativum-sprayed plants significantly reduced the number of T. absoluta-infested leaflets and did not affect the establishment and reproduction of N. tenuis. Additionally, the spraying of A. millefolium and A. sativum increased the expression of defense genes in the plants, triggering the release of herbivory-induced plant volatiles (HIPVs), such as C6 green leaf volatiles, monoterpenes, and aldehydes, which can be messengers in tritrophic interactions. The results suggest that PEOs from A. millefolium and A. sativum can provide a dual benefit for controlling arthropod pests, as they can directly exhibit toxicity against these pests while also activating plant defense mechanisms. Overall, this study provides new insights into using PEOs as a sustainable solution for controlling pests and diseases in agriculture, by reducing synthetic pesticides and promoting the use of natural predators.

Control of Leiodes cinnamomeus (Coleoptera: Leiodidae) in Cultivated Black Truffle Orchards by Kairomone-Based Mass Trapping.

The monoculture situation of truffle cultivation is favoring the appearance of pests that would not be economically important in naturally balanced forest ecosystems. The most prominent of them is the European truffle beetle Leiodes cinnamomeus (Panzer) (Coleoptera: Leiodidae), for which there are no effective control methods capable of reducing its populations. The potential of the mass trapping technique against this beetle, based on adapted pitfall traps and the semiochemical methyl disulfide as an attractant, is explored in the present work. Two trap densities (40 and 80 traps/ha) were tested in 2-yr field trials carried out in the region of Teruel (Spain) with black truffle cultivation tradition. Kairomone dispensers were placed in the field immediately before adult outbreak and remained active there throughout the season. The efficacy of each treatment was measured according to the reduction in beetle populations and the damaged truffles in the center of the treated areas. The results showed that both trapping densities reduced adult populations (mean 57% catch reduction), but 80 traps/ha were needed to significantly lower damage parameters (>40% reduction), percentage of attacked truffles and number of galleries/g truffle. The cost effectiveness of these treatments and possible improvements are discussed.

Production of Volatile Moth Sex Pheromones in Transgenic Nicotiana benthamiana Plants.

Plant-based bioproduction of insect sex pheromones has been proposed as an innovative strategy to increase the sustainability of pest control in agriculture. Here, we describe the engineering of transgenic plants producing (Z)-11-hexadecenol (Z11-16OH) and (Z)-11-hexadecenyl acetate (Z11-16OAc), two main volatile components in many Lepidoptera sex pheromone blends. We assembled multigene DNA constructs encoding the pheromone biosynthetic pathway and stably transformed them into Nicotiana benthamiana plants. The constructs contained the Amyelois transitella AtrΔ11 desaturase gene, the Helicoverpa armigera fatty acyl reductase HarFAR gene, and the Euonymus alatus diacylglycerol acetyltransferase EaDAct gene in different configurations. All the pheromone-producing plants showed dwarf phenotypes, the severity of which correlated with pheromone levels. All but one of the recovered lines produced high levels of Z11-16OH, but very low levels of Z11-16OAc, probably as a result of recurrent truncations at the level of the EaDAct gene. Only one plant line (SxPv1.2) was recovered that harboured an intact pheromone pathway and which produced moderate levels of Z11-16OAc (11.8 µg g-1 FW) and high levels of Z11-16OH (111.4 µg g-1). Z11-16OAc production was accompanied in SxPv1.2 by a partial recovery of the dwarf phenotype. SxPv1.2 was used to estimate the rates of volatile pheromone release, which resulted in 8.48 ng g-1 FW per day for Z11-16OH and 9.44 ng g-1 FW per day for Z11-16OAc. Our results suggest that pheromone release acts as a limiting factor in pheromone biodispenser strategies and establish a roadmap for biotechnological improvements.

Airborne Pheromone Quantification in Treated Vineyards with Different Mating Disruption Dispensers against Lobesia botrana.

Mating disruption (MD) is widely used against the European grapevine moth (EGVM), Lobesia botrana (Denis and Schiffermüller; Lepidoptera: Tortricidae), by installing passive dispensers or aerosol devices. The present work reports a new sampling and quantification methodology to obtain absolute data about field airborne pheromone concentration based on air samplings and sensitive chromatographic-spectroscopic methods. Samplings were performed in fields treated with passive dispensers or aerosol devices at different moments throughout the crop cycle to study how they act and how the disruption is triggered. Moreover, pheromone adsorption and releasing capacity of vine leaves were studied to elucidate their role in the disruption. Although both types of dispensers were effective in limiting the damage inflicted by EGVM, they performed differently and provided different airborne pheromone concentration profiles. Results also proved that leaves were able to adsorb and release part of the airborne pheromone acting as subsequent and additional pheromone sources. This fact could explain the different concentration profiles. Moreover, our results suggest that lower pheromone emission than that of the current passive dispensers still could provide an adequate performance in the field. Competitive mechanisms involved in MD using both dispensers, the dynamics of the airborne pheromone throughout the time and the importance of the canopy are discussed.

Study on the optimum pheromone release rate for attraction of Chilo suppressalis (Lepidoptera: Pyralidae).

Traditional chemical control against Chilo suppressalis Walker is currently being replaced in Spain by new methods based on pheromones. A key step to improve the efficacy of these methods is the determination of the optimum pheromone release rate, which is still uncertain for this pest. In this work, the pheromone release profile and the field performance of a new mesoporous dispenser was compared with a standard commercial dispenser. For this purpose, pheromone loads were extracted from field-aged dispensers and quantified by gas chromatography with flame ionization detector. In addition, a field trial was carried out with traps baited with one, two, or three mesoporous dispensers per trap, as well as with traps containing one standard dispenser. We found that the highest number of field catches did not correspond to the highest pheromone emission rate, which suggests a repellent effect of the insect if the emission is excessive. The results suggest that the attractant activity was maximized by emitting approximately 34 microg/d. The efficacy of the mesoporous dispenser and its possible improvements are discussed.

Sex Pheromone of the Invasive Mealybug Citrus Pest, Delottococcus aberiae (Hemiptera: Pseudococcidae). A New Monoterpenoid with a Necrodane Skeleton.

Native to sub-Saharan Africa, Delottococcus aberiae De Lotto (Hemiptera: Pseudoccidae) is an invasive mealybug that has been recently reported in Europe, seriously damaging citrus production in eastern Spain. In this study, we isolated and determined the structure of the sex pheromone of D. aberiae, to provide a highly specific and effective lure for detecting, monitoring, and potentially controlling this pest. The volatile profile of D. aberiae virgin and mated females was studied by aeration and collection of effluvia in Porapak-Q. The resulting extracts were analyzed by gas chromatography coupled to mass spectrometry (GC-MS), revealing a candidate compound specific of virgin females. GC-MS and nuclear magnetic resonance spectroscopy data evidenced a new compound, (4,5,5-trimethyl-3-methylenecyclopent-1-en-1-yl)methyl acetate, with an unusual ß-necrodol skeleton. This compound was synthesized and shown to be attractive to male D. aberiae in both laboratory and field experiments. A GC analysis using an enantioselective stationary phase and polarimetry analyses of the synthetic enantiomers showed the natural compound emitted by virgin females to be the - enantiomer.

Survey on Drosophila suzukii Natural Short-Term Dispersal Capacities Using the Mark-Release-Recapture Technique.

Spotted wing drosophila, Drosophila suzukii Matsumura (Diptera: Drosophilidae), has become a key pest for soft fruits and cherries in Europe in less than a decade since the first outbreak in 2007. Although this pest's passive dispersal ability has been observed over more than 1400 km in 1 year, active spread has not yet been extensively studied. A mark-release-recapture (MRR) method based on protein-marked flies was employed to determine the flight capacity of D. suzukii. Sterile marked flies were released and recaptured in a trap grid at increasing distances from 10 to 250 m from the releasing point to study flight distance during periods ranging from 3 h to 1 week. MRR experiments were replicated in the presence and absence of host fruits to study how they could affect dispersal behavior. The dispersal capacity of the Mediterranean fruit fly, Ceratitis capitata Wiedemann (Diptera: Tephritidae) was also studied under the same conditions. The results showed a low dispersal ability for D. suzukii, with a daily flight distance below 100 m with no predominant wind. The implications on natural dispersion and control methods based on attractants are discussed.

Improvements in Rhynchophorus ferrugineus (Coleoptera: Dryophthoridae) Trapping Systems.

Improved trap efficacy is crucial for implementing control methods for red palm weevil, Rhynchophorus ferrugineus (Olivier; Coleoptera: Dryophthoridae), based on trapping systems, such as mass trapping, attract and infect or attract and sterilize techniques. Although new trap designs have been proposed and aggregation pheromone dispensers have been optimized, aspects such as the use of co-attractants (molasses) and trap placement are still not well defined and standardized. The efficacy of three concentrations of molasses and different formulations to reduce water evaporation in traps was studied in different field trials to improve trapping systems and to prolong trap servicing periods. In addition, the performance of installing groups of traps or single traps was also evaluated with the aim of improving the attracted/captured weevils ratio. Our results showed that captures increased when molasses were added at 15% to the water contained in the trap and that a thin layer of oil, created by adding 2-3% of paraffinic oil to water, was able to effectively reduce evaporation and prolong trap servicing periods. Moreover, 3.5-fold more weevils were captured when placing five traps instead of one at the same trapping point. Results obtained allow improved efficacy and may have an impact in the economic viability of trapping systems and, therefore, in integrated pest management programs.

Development of an attract-and-infect system to control Rhynchophorus ferrugineus with the entomopathogenic fungus Beauveria bassiana.

BACKGROUND: A new Beauveria bassiana-based attract and infect device (AID) to control Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae) was developed. The virulence and persistence of the fungal formulation used in the AID were evaluated in the laboratory. Semi-field and field trials were carried out to validate the results and establish the potential of this device as a control tool. RESULTS: In laboratory conditions, a 50% lethal time (LT50 ) of 4.33 days was obtained when adults (7-10 days old) were exposed to the inoculation tunnel (IT) containing 1 × 1010 conidia g-1 in an oil-based fungal formulation. This formulation maintained conidium viability at 50% for up to 2 months. Moreover, when adults were exposed to 2.5-month field-aged ITs, mortality still reached 50% 40 days after exposure. In addition, no differences were observed between ITs aged in early spring and those aged in summer, suggesting that the fungal formulation is not strongly affected by environmental factors in Mediterranean basin conditions. Semi-field assays showed that the device allowed an easy transit of weevils through the IT, which were effectively attracted and infected. Using the AIDs in 4-ha plot field trials, a reduction of >50% in the percentage of infested sentinel palms was obtained. CONCLUSION: Based on the results obtained in terms of the efficacy and persistence of this new AID in the field and its potential in reducing R. ferrugineus populations and palm infestation, this device could become a key tool for the management of R. ferrugineus. © 2018 Society of Chemical Industry.

Identification of the Male-Produced Aggregation Pheromone of the Four-Spotted Coconut Weevil, Diocalandra frumenti.

The four-spotted coconut weevil, Diocalandra frumenti Fabricius (Coleoptera: Dryophthoridae), is a small weevil found attacking economically important palm species, such as coconut, date, oil, and Canary palms. Given the scarcity of detection and management tools for this pest, the availability of a pheromone to be included in trapping protocols would be a crucial advantage. Previous laboratory experiments showed evidence for aggregation behavior; thus, our main goal was to identify the aggregation pheromone in this species. The volatile profile of D. frumenti individuals was studied by aeration and collection of effluvia in Porapak-Q and also by solid phase microextraction (SPME) techniques. Moreover, solvent extraction of previously frozen crushed individuals was also performed. All resulting extracts and SPME fibers were analyzed by gas chromatography coupled to mass spectrometry (GC-MS). The comparison of male and female samples provided the candidate compound, 5-ethyl-2,4-dimethyl-6,8-dioxabicyclo[3.2.1]octane (multistriatin), whose biological activity was evaluated in olfactometer and field assays.

Lures for red palm weevil trapping systems: aggregation pheromone and synthetic kairomone.

BACKGROUND: The optimisation of the lure is essential for the implementation of trapping systems to control insect pests. In this work, the response of the red palm weevil (RPW), Rhynchophorus ferrugineus Olivier, to increasing emission rates of its aggregation pheromone (ferrugineol) and the efficacy of a convenient synthetic kairomone based on fermentation odours (ethyl acetate and ethanol) have been evaluated in different years and locations along the Mediterranean basin. RESULTS: In general, although capture data and emission had noticeable variability among locations, significantly fewer RPW were captured in pyramidal Picusan® traps with the lowest ferrugineol emission rates tested (0.6-3.8 mg day-1 ). Captures increased rapidly with ferrugineol emission up to 4-5 mg day-1 ; then, higher emission rates did not improve or reduce captures, up to the highest emission rate tested of 50.9 mg day-1 . Thus, there is no evidence of an optimum release rate corresponding to a maximum of RPW catches. Traps baited with the synthetic kairomone (1:3 ethyl acetate/ethanol) captured 1.4-2.2 times more total weevils than traps baited only with ferrugineol. Moreover, in most of the locations, the synthetic blend was at least as effective as the local coattractants used (plant material + molasses). CONCLUSIONS: Ferrugineol emission rate can vary in a wide range without significantly affecting RPW response. Coattractants based on fermenting compounds, ethyl acetate and ethanol, are able to improve the attractant level of ferrugineol and could be employed to replace non-standardised natural kairomones in RPW trapping systems after further optimisation of their proportions and doses. © 2016 Society of Chemical Industry.

Bait station devices can improve mass trapping performance for the control of the Mediterranean fruit fly.

BACKGROUND: The use of traps and other attract-and-kill devices in pest management strategies to reduce Mediterranean fruit fly populations has proved to be efficient. Nevertheless, many farmers are concerned about the effect of these devices on the trees where they are hung. Direct field observations have revealed that fruit damage is higher in trees with traps than in trees without them. This work evaluates the efficacy of different types of attract-and-kill device to protect fruit of the single tree on which the device is placed. RESULTS: Results suggested that trees with traps had at least the same fruit damage than trees without them. When traps were baited with protein hydrolysate, fruit damage was even higher than in trees without traps. However, fruit damage was significantly diminished when efficient bait station devices were used. CONCLUSION: Although mass trapping is able to control fruit fly populations as a control method, trees with some types of trap and bait are more susceptible to fly puncture. However, bait station devices reduce fruit damage in the single trees where they are hung. Bait stations are more efficient in fruit protection because fruit flies are affected as soon as they contact the device. Some recommendations for the use of the different attract-and-kill devices are discussed.

Role of α-copaene in the susceptibility of olive fruits to Bactrocera oleae (Rossi).

The influence of α-copaene as a fruit volatile in the susceptibility of Olea europaea L. to the olive fruit fly Bactrocera oleae (Rossi) has been investigated. By studies on the relative area of volatile components from different cultivars, a positive correlation was found between the abundance of α-copaene in the samples and the corresponding degree of fruit infestation. SPME-GC-MS analysis of volatiles from uninfested fruits of O. europaea L. cv. Serrana were performed over two years to determine the variation of α-copaene throughout the different phenological stages. The results suggested that this sesquiterpene has a significant effect on cultivar susceptibility and may act as an oviposition promoter. Further analysis by chiral GC showed that olive fruits release both α-copaene enantiomers. Bioassays on each enantiomer revealed that fruits with increased amounts of (+)-α-copaene favor oviposition of B. oleae females, whereas the increase of (-)-α-copaene affords no statistically significant differences in host preference.

Identification of pheromone synergists for Rhynchophorus ferrugineus trapping systems from Phoenix canariensis palm volatiles.

Trapping systems for the red palm weevil, Rhynchophorus ferrugineus Olivier, rely on the use of natural plant odor sources to boost the attractiveness of the aggregation pheromone. The identification of the key odorants involved in attraction is essential in the development of a synthetic pheromone synergist to replace the nonstandardized use of plant material in traps. Canary Islands date palms (Phoenix canariensis) have become preferred hosts for R. ferrugineus in Europe; thus, the volatile profile of different P. canariensis plant materials, including healthy and infested tissues, is investigated in the present work by means of solid phase microextraction (SPME-GC-MS), aimed to identify pheromone synergists. The electroantennography (EAG) response of the compounds identified was recorded, as well as the preliminary field response of several EAG-active compounds. The so-called "palm esters" (ethyl acetate, ethyl propionate, ethyl butyrate, and propyl butyrate) elicit the strongest EAG responses but performed poorly in the field. Mixtures of esters and alcohols give evidence of better performance, but release rates need further optimization.