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In spite of its incidence on pistachio trees, the chemical ecology of Labidostomis lusitanica (Germar) (Coleoptera: Chrysomelidae) has been neglected so far. In this work we provide the first evidence of a biologically active male-specific compound that may be promoting field aggregation. Headspace collections by solid-phase microextraction from feral males and females reported the presence of 2-isobutyl-3-methoxypyrazine exclusively on males. Electroantennographic recordings revealed that males and females responded in a dose-dependent manner to increasing stimuli of 2-isobutyl-3-methoxypyrazine, with females overall displaying a higher response than males. In dual-choice tests, both males and females showed a significant preference for the compound in comparison to a pure air stimulus. In the light of these results, the possible role of 2-isobutyl-3-methoxypyrazine as an aggregation cue in L. lusitanica is discussed.

Simple Summary The flathead oak borer Coraebus undatus (F.) (Coleoptera: Buprestidae) is a major pest of cork oak (Quercus suber) along the Mediterranean Basin that generates significant economic losses in the cork industry. Larvae bore long galleries and feed on the cork generating layer, thus affecting its quality. At present, the semiochemistry of this species is poorly known, and therefore the elucidation of chemicals involved in its intraspecific communication may allow the development of novel control tools. We determined that both sexes release the compound 1,7-dioxaspiro[5.5]undecane, and the biological activity of the compound was addressed by means of electroantennography and behavioral assays. The attractiveness of the compound on both sexes under laboratory conditions contrasts to its performance in field trials, which may be explained by features inherent to the methodological design (e.g., the absence of a contextually related visual stimulus or trap deployment height). This is the first time in which an intraspecific compound has been reported as attractive for the species, and practical implications for the assessment of its activity under natural conditions are also further discussed. Abstract The main aim of this work was to identify semiochemicals from the jewel beetle Coraebus undatus (F.) (Coleoptera: Buprestidae) that may aid in the improvement of current monitoring tools. First, HS-SPME collections revealed that individually sampled adults (>7 days old) of both sexes release the spiroacetal 1,7-dioxaspiro[5.5]undecane (olean). Electroantennographic recordings from both sexes exposed to increasing amounts of olean followed a dose-dependent pattern, with females being more responsive than males to the highest amount of the compound (100 µg). In double-choice assays, adults older than seven days were significantly attracted to olean, whereas this attraction was not detected in insects aged less than seven days. Indeed, a repellent effect was observed in young females. Subsequent field trials employing sticky purple prism traps revealed that there were no differences among the number of insects caught in control and olean-baited traps at two different release rates (0.75 and 3.75 mg/day). Interestingly, all the trapped specimens were determined as mated females, regardless of the presence of olean. Overall, these findings provide a basis for unraveling the chemical ecology of the species, although further research is still needed to determine the specific role of this compound within the chemical communication of the species.

The main aim of this work was to identify semiochemicals from the jewel beetle Coraebus undatus (F.) (Coleoptera: Buprestidae) that may aid in the improvement of current monitoring tools. First, HS-SPME collections revealed that individually sampled adults (>7 days old) of both sexes release the spiroacetal 1,7-dioxaspiro[5.5]undecane (olean). Electroantennographic recordings from both sexes exposed to increasing amounts of olean followed a dose-dependent pattern, with females being more responsive than males to the highest amount of the compound (100 µg). In double-choice assays, adults older than seven days were significantly attracted to olean, whereas this attraction was not detected in insects aged less than seven days. Indeed, a repellent effect was observed in young females. Subsequent field trials employing sticky purple prism traps revealed that there were no differences among the number of insects caught in control and olean-baited traps at two different release rates (0.75 and 3.75 mg/day). Interestingly, all the trapped specimens were determined as mated females, regardless of the presence of olean. Overall, these findings provide a basis for unraveling the chemical ecology of the species, although further research is still needed to determine the specific role of this compound within the chemical communication of the species., This research was funded by Departament d’Agricultura, Ramaderia, Pesca i Alimentació, Generalitat de Catalunya (AG-2019-20061).

Plant essential oils are considered as important bio-sources for the development of natural and environmentally safe pest control tools due to their multiple modes of action on insects. In this paper we have evaluated the activity of commercially available thyme oil and its constituents thymol, carvacrol, and p-cymene, as potential disruptants of the pheromone-mediated communication in the major pest moths Spodoptera littoralis Boisduval (Lepidoptera: Noctuidae), and Grapholita molesta (Busck) (Lepidoptera: Tortricidae). In electroantennographic assays, the antennal response of males to thyme oil, thymol, and carvacrol was altered at high doses (103–104 µg), shifting the signal waveform into a biphasic negative–positive potential that caused a decay in the response. In wind tunnel assays, pheromone-mediated attraction of males of both species was interrupted in presence of thyme oil. Further trials demonstrated that thymol alone reduced the number of G. molesta and S. littoralis males landing on the pheromone source. This effect did not differ from that of thyme oil, although the latter provoked a significant reduction on downwind behavior steps in S. littoralis. Overall, our findings provide a preliminary basis for delving into the effect of thyme oil, and especially of its major constituent thymol, as potential mating disruptants of both species., We gratefully acknowledge MINECO for an FPI contract to A.D. This work was supported by project AGL2012-39869-C02-01 from MINECO with assistance from the European Regional Development Fund for financial support.

Jewel beetles are colorful insects, which use vision to recognize their conspecifics and can be lured with colored traps. We investigated the retina and coloration of one member of this family, the flathead oak borer

During the last few decades, extensive efforts has been made to design nanocarriers to transport drugs into the central nervous system (CNS). However, its efficacy is limited due to the presence of the Blood-Brain Barrier (BBB) which greatly reduces drug penetration making Drug Delivery Systems (DDS) necessary. Polymeric nanoparticles (NPs) have been reported to be appropriate for this purpose and in particular, poly(lactic-co-glycolic acid) (PLGA) has been used for its ability to entrap small molecule drugs with great efficiency and the ease with which it functionalizes NPs. Despite the fact that their synthetic identity has been studied in depth, the biological identity of such manufactured polymers still remains unknown as does their biodistribution and in vivo fate. This biological identity is a result of their interaction with blood proteins, the so-called “protein corona” which tends to alter the behavior of polymeric nanoparticles in the body. The aim of the present research is to identify the proteins bounded to polymeric nanoparticles designed to selectively interact with the BBB. For this purpose, four different PLGA NPs were prepared and analyzed: (i) “PLGA@Drug,” in which a model drug was encapsulated in its core; (ii) “8D3-PLGA” NPs where the PLGA surface was functionalized with a monoclonal anti-transferrin receptor antibody (8D3 mAb) in order to specifically target the BBB; (iii) “8D3-PLGA@Drug” in which the PLGA@Drug surface was functionalized using the same antibody described above and (iv) bare PLGA NPs which were used as a control. Once the anticipated protein corona NPs were obtained, proteins decorating both bare and functionalized PLGA NPs were isolated and analyzed. Apart from the indistinct interaction with PLGA NPs with the most abundant serum proteins, specific proteins could also be identified in the case of functionalized PLGA NPs. These findings may provide valuable insight into designing novel vehicles based on PLGA NPs for crossing the BBB., The authors are grateful to the Agencia Estatal de Investigación (AEI) and Fondo Europeo de Desarrollo Regional (FEDER) for funding (Project CTQ2017-84998-P) and the Instituto de Salud Carlos III (ISCIII) (CB06/01/1058). CIBER BBN is an initiative funded by the VI National R+D+I Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program, CIBER actions and financed by ISCIII with assistance from the European Regional Development Fund. The authors want to thank Dr. Miriam Royo’s research group and Dr. Gerardo Acosta for their valuable assistance in HPLC experiments and Ms. Suzanne Córdova for revising and correcting this document. The authors also want to thank NANBIOSIS ICTS and the Nanostructured Liquid Characterization Unit (Unit U12) for DLS measurements. The authors also acknowledge the “Grupo de Nanotecnología Farmacéutica”, from the University of Barcelona (UB), in the Faculty of Pharmacy and Food Sciences, which forms an R&D Associated Unit to CSIC.

The tomato leafminer Tuta absoluta Meyrick (Lepidoptera: Gelechiidae) is one of the most important pests of tomato worldwide. However, in spite of its tremendous economic importance, the success of environmentally friendly measures to control the pest is still limited. Study of physiological and behavioral parameters that affect pheromone production has provided useful information for pest management. Our results show no clear difference in pheromone production by females over the period from 2 h before to 2 h after the scotophase. However, pheromone production was clearly dependent on female age, with young females producing the highest amount of each pheromone component 10 days after emergence. In the presence of the host plant (physical contact and olfaction of the plant volatiles), virgin and mated females produced higher amounts of the major component of the pheromone (TDTA) than those in the absence of plant and those devoid of olfaction (antennectomized) but in physical contact with the plant. In electrophysiological experiments, TDTA elicited slightly lower responses on male antennae than the pheromone mixture. When stimulated at certain time intervals after the first exposure to TDTA, male antennae became more sensitive to the stimulus (sensitization effect). For the first time in an insect of the family Gelechiidae, we have found that females are able to detect their own pheromone (autodetection). Altogether, our results may represent a step forward in the knowledge of the chemical communication of this important pest.

© 2019 by the authors., The tomato leafminer Tuta absoluta Meyrick (Lepidoptera: Gelechiidae) is one of the most important pests of tomato worldwide. However, in spite of its tremendous economic importance, the success of environmentally friendly measures to control the pest is still limited. Study of physiological and behavioral parameters that affect pheromone production has provided useful information for pest management. Our results show no clear difference in pheromone production by females over the period from 2 h before to 2 h after the scotophase. However, pheromone production was clearly dependent on female age, with young females producing the highest amount of each pheromone component 10 days after emergence. In the presence of the host plant (physical contact and olfaction of the plant volatiles), virgin and mated females produced higher amounts of the major component of the pheromone (TDTA) than those in the absence of plant and those devoid of olfaction (antennectomized) but in physical contact with the plant. In electrophysiological experiments, TDTA elicited slightly lower responses on male antennae than the pheromone mixture. When stimulated at certain time intervals after the first exposure to TDTA, male antennae became more sensitive to the stimulus (sensitization effect). For the first time in an insect of the family Gelechiidae, we have found that females are able to detect their own pheromone (autodetection). Altogether, our results may represent a step forward in the knowledge of the chemical communication of this important pest., This research was funded by MINECO, project AGL2015-66469-R with assistance from the European Regional Development Fund

Degradation of solid polymers in the aquatic environment encompasses a variety of biotic and abiotic processes giving rise to heterogeneous patterns across the surface of the material, which cannot be investigated using conventional Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) that only renders an "average" picture of the sample. In that context, MALDI-TOF MS Imaging (MALDI MSI) provides a rapid and efficient tool to study 2D spatial changes occurred in the chemical composition of the polymer surface.Commercial polycaprolactone diol (average molecular weight of 1250 Da) was selected as test material because it had been previously known to be amenable to biological degradation. The test oligomer probe was incubated under aerobic and denitrifying conditions using synthetic water and denitrifying mixed liquor obtained from a wastewater treatment plant respectively. After ca. seven days of exposure the mass spectra obtained by MALDI MSI showed the occurrence of chemical modifications in the sample surface. Observed heterogeneity across the probe's surface indicated significant degradation and suggested the contribution of biotic processes. The results were investigated using different image processing tools. Major changes on the oligomer surface were observed when exposed to denitrifying conditions. © 2016., This study has been financially supported by the EU through the FP7 project GLOBAQUA (Grant agreement No 603629 ), and by the Generalitat de Catalunya (Consolidated Research Groups “2014 SGR 418—Water and Soil Quality Unit” and "2014 SGR 291—ICRA"). It reflects only the author's views. The Community is not liable for any use that may be made of the information contained therein. The authors are thankful to Dr. P. Eichhorn for his critical reading of the manuscript and the valuable comments provided and to Maria Casellas (Experimental Streams Facility, Catalan Institute for Water Research, Girona, Spain) for her technical help in the operation of the artificial stream. Appendix A

The tomato leafminer, Tuta absoluta, is one of the major pests of tomato and other Solanaceae in many regions worldwide. In the search for new strategies to control this pest, we present herewith the stereoselective synthesis, electrophysiological activity, functional analysis, and field tests of new chemicals as possible antagonists of the sex pheromone of the leafminer. The chemicals are methyl ketone (MK) and trifluoromethyl ketone (TFMK) structural analogues of both components of the pheromone. Most of the chemicals exerted per se some electrophysiological activity and inhibited the electroantennographic response to the pheromone when vapors of the inhibitor were passed over the antennae. Except TFMK 3, which elicited a modest effect, the compounds did not exhibit antiesterase activity on the pheromone-degrading enzymes of the antennae, but in the field the chemicals, particularly MK 5, notably decreased the number of catches when mixed with the pheromone in 1:1 and 10:1 ratios, regardless the infestation level of the plot. These results suggest that MK 5 is a good behavioral antagonist of the pheromone to be considered as a putative agent to control the pest in new future integrated pest management (IPM) strategies.

In insects, the olfactory system displays a high degree of plasticity. In Spodoptera littoralis, pre-exposure of males to the sex pheromone has been shown to increase the sensitivity of the olfactory sensory neurons at peripheral level. In this study, we have investigated this sensitization effect by recording the electroantennographic responses of male antennae to the major sex pheromone component (Z,E)-9,11-tetradecadienyl acetate and to the minor components (Z,E)-9,12-tetradecadienyl acetate and (Z)-9-tetradecenyl acetate. Responses to the conjugated diene acetate at 1 and 10 µg and to the unconjugated ester at 10 µg at three different times (11, 22 and 33 min) after pre-exposure (T = 0 min) were significantly higher than those at T = 0, whereas no increase of sensitivity to the pheromone was elicited by any dose of the minor monoene acetate. In addition, pre-exposed antennae to sub-threshold amounts (0.1, 1 and 10 ng) of the major pheromone component also induced an increased response to the chemical at different times (5 and 15 min) after exposure. Our results revealed that pre-exposed isolated antennae display a short-term higher sensitivity at the peripheral level when compared to naive antennae. In addition, we provide evidence of a peripheral sensitization mediated not only by the major pheromone component, but also by the minor unconjugated diene acetate, and the induction of this sensitivity appears to be dependent on the pre-exposure dose and the time span between pre-exposure and subsequent recordings. Possible implications of the sensitization effect displayed by the minor component for a more effective discrimination of the pheromone bouquets of other closely related species are highlighted.

The flathead oak borer Coroebus undatus F. (Coleoptera: Buprestidae) is one of the primary pests of cork oak Quercus suber L. in the Mediterranean region causing great economic losses to the cork industry. Very little is known about its biology and behavior and, so far, no control measures have been established. We present the results of a pilot study aimed to develop an efficient trapping method for monitoring this harmful pest. In a 3-year field study, purple-colored prism traps baited with a mixture of green leaf volatiles (GLVs) from the host have been shown the most effective combination to catch C. undatus adults (solely females) compared to other trap and lure types tested. Wavelength and reflectance measurements revealed that purple traps exhibit reflectance peak values similar to those found in the abdominal and elytral cuticle of both sexes, suggesting the involvement of visual cues for mate location in this species. The data presented are the first to demonstrate captures of adults of the genus Coroebus by an attractant-based trapping method.

The genus Spodoptera contains some of the most destructive crop pests in the world owing to the worldwide distribution of the different species and their wide host ranges. Uses of semiochemicals for insect control have been recently reviewed, but strategies developed specifically against Spodoptera species are scarce. In this review, we present an updated account of the semiochemicals and other natural product-based approaches to monitor and control the most damaging Spodoptera spp. In general, successful control of Spodoptera littoralis, S. frugiperda, S. litura, and S. exigua have been reported through mass trapping, mating disruption, and attract-and-kill methods. An updated survey of the pheromone components of these species and an outlook of future perspectives against these pests are also outlined.

Outbreaks of locusts cause enormous economic losses to agriculture in many countries. To develop environmentally friendly strategies for their control, much research has been focused on the factors that influence locust biology, particularly infochemical-mediated interactions. We present herein the identification and synthesis of both Z and E isomers of phytal (3,7,11,15-tetramethylhexadec-2-enal, 1), which are involved in chemical communication and behaviour of the Moroccan locust, Dociostaurus maroccanus, a serious agricultural pest. The compound was identified by comparison of its chromatographic and spectrometric features and microchemical reactions with those of a synthetic sample. The natural compound was shown to have the R,R configuration by chiral HPLC analysis, and its structure is unique as an insect pheromone component. Both isomers of phytal are produced by sexually mature adult males and elicit electroantennographic responses in antennae of both sexes. In two-choice olfactometer bioassays, males and females significantly preferred the stream enriched with racemic phytal to the control. In contrast, hydrogenated phytal was behaviourally inert. Both isomers of phytal are specific to D. maroccanus as they are absent in the closely related, habitat-sharing species Dociostaurus jagoi and Calliptamus wattenwylianus. Legs and wings are the main release sites of the compound: approximately 90 % of that emitted by living individuals. In biosynthetic studies, phytal appears to proceed from oxidation of phytol (2) after injection of deuterated phytol into the abdomen of the insect or after administration in the diet. Our results demonstrate that phytal is a candidate sex pheromone component of the Moroccan locust; it is produced by mature males, and might be eavesdropped upon by conspecific males.

Characterization of river ecosystems must take into consideration both structural and functional aspects. For the latter, a convenient and simple approach for routine monitoring is based on the decomposition of organic matter measured in terms of breakdown of natural organic substrates like leaf litter, wood sticks. Here we extended the method to a synthetic organic material using polymer probes characterized by MALDI-TOF/MS. We first characterized several commercial available polymers, and finally selected polycaprolactonediol 1250 (PCP 1250), a polyester oligomer, as the most convenient for further studies. PCP 1250 was first tested at mesocosms scale under conditions simulating those of the river, with and without nutrient addition for up to 4 weeks. Differences to the starting material measured in terms of changes in the relative ion peak intensities were clearly observed. Ions exhibited a different pattern evolution along time depending on their mass. Greatest changes were observed at longest exposure time and in the nutrient addition treatment. At shorter times, the effect of nutrients (addition or not) was indistinguishable. Finally, we performed an experiment in 11 tributaries of the Ebro River during 97 days of exposure. Principal Component Analysis confirmed the different behavior of ions, which were clustered according to their mass. Exposed samples were clearly different to the standard starting material, but could not be well distinguished among each other. Polymer mass loss rates, as well as some environmental variables such as conductivity, temperature and flow were correlated with some peak intensities. Overall, the interpretation of field results in terms of environmental conditions remains elusive, due to the influence of multiple concurrent factors. Nevertheless, breakdown of synthetic polymers opens an interesting field of research, which can complement more traditional breakdown studies to assess river ecosystem functioning. © 2016 The Authors, This study has been financially supported by the EU through the FP7 project GLOBAQUA (Grant agreement No 603629 ), and by the Generalitat de Catalunya (Consolidated Research Groups “ 2014 SGR 418 —Water and Soil Quality Unit” and 2014 SGR 291 —ICRA). It reflects only the author's views. The Community is not liable for any use that may be made of the information contained therein. The authors are thankful to Maria Casellas (Experimental Streams Facility, Catalan Institute for Water Research, Girona, Spain) for her technical help in the operation of the artificial streams. Appendix A

Department of Education, Universities and Research of the Basque Country Department of Agriculture and Fisheries of the Basque Country

There is currently relatively little information on how marine organisms process and analyze sound, making assessments about the impacts of artificial sound sources in the marine environment difficult. However, such assessments have become a priority because noise is now considered as a source of pollution that increasingly affects the natural balance of marine ecosystems. We present the first morphological and ultrastructural evidence of massive acoustic trauma, not compatible with life, in four cephalopod species subjected to low-frequency controlled-exposure experiments. Exposure to low-frequency sounds resulted in permanent and substantial alterations of the sensory hair cells of the statocysts, the structures responsible for the animals' sense of balance and position. These results indicate a need for further environmental regulation of human activities that introduce high-intensity, low-frequency sounds in the world's oceans.

The pheromone composition of the Spanish population of the beet armyworm (BAW), Spodoptera exigua (Lepidoptera: Noctuidae), was identified. Analysis of female gland extracts showed the presence of compounds Z9,E12–14:Ac (1), Z9–14:Ac (2), Z11–16:Ac (3), Z9,E12–14:OH (4), Z9–14:OH (5), and Z11–16:OH (6) in a ratio of 26:11:1:22:31:9. The amount of compound per gland ranged from 2.08 ng for 5 to 0.09 ng for 3. However, analysis of female volatiles by SPME revealed only the presence of compounds, 1, 2, 3, and 5 in a 34:40:4:22 ratio. In electroantennogram assays, compound 1 elicited the highest response, and the C14 acetates evoked higher electrophysiological responses than the corresponding alcohols or C16 isomers. In a wind tunnel, no behavioral difference was observed between formulations based on the gland extracts and female volatiles. In both cases, males responded as when virgin females were used as the attractant source. Compound 1 alone elicited upwind flight by males, but required the presence of compound 5 in a 80:20 to 40:60 ratio for full activity. Ternary mixtures of 1, 5 and the minor components did not improve the performance of the 1+5 blend in a 60:40 ratio. In the field, the mixture 1+5+3 in a 56:37:7 ratio was the most attractive formulation, and is expected to be useful in future pest control strategies., We gratefully acknowledge T. Cabello (University of Almería) for providing us with BAW larvae for rearing and useful advice, P. Fuchs (Kenogard, S.A.) for conducting the field tests, and SEDQ S.A. for providing us with compounds 3–6. We are also indebted to MICINN for an I3P fellowship to P. Acín and a Ramón y Cajal contract to C.Q. Financial support was provided by CICYT (project AGL 2006-13489-C02-01/AGR).

Olfaction is essential for processing chemical signals in insects, but characterizing the proteins implicated in this process has proved challenging. We optimized 2-DE gel resolution of insect proteins by using a buffer containing two reducing agents, DTT and hydroxyethyl disulfide. This buffer clearly improved resolution and decreased spot streaking and spot trains of 2-DE in comparison to DTT alone. We described for the first time that the buffer with DTT and hydroxyethyl disulfide further to reducing streaking in the basic part of the gel eliminates false spots in the acidic gel regions that appeared when only DTT was used as reducing agent.

Most insect pheromones comprise multicomponent blends of geometric or optical isomers, and one major question is how insects produce species-specific ratios of components for successful reproductive isolation. Key enzymes suggested to be involved in pheromone biosynthesis are acetyl-coenzyme A carboxylase and fatty acyl synthetase, chain-shortening enzymes, desaturases, elongases, reductases, oxidases, and alcohol acetyl transferases. The female pheromone composition of the Egyptian armyworm Spodoptera littoralis (Boisd.) is highly dependent on the origin of the strain. In this review, we present a summary of the different reported pheromone compositions of the moth, including from our recent studies on this subject, as well as the biosynthetic routes to the different components and the molecular approaches involved. In addition, the key role played in the proposed biosynthetic pathways by a number of important biosynthetic enzymes, such as chain shortening enzymes, desaturases and alcohol acetyl transferases, is outlined, as well as the latest developments on the inhibition of these enzymes.

We present herein a review article of the latest developments of the biorational approaches in pest management appeared in the literature from 1997 to date. The proposed advantages of the biopesticides including their specificity, safety to non-target organisms, particularly mammals, and utilization in low, sometimes minute, amounts have led to an intensive research program by public and private institutions resulting in an avalanche of reports in attempts to discover and develop newer and safer pesticides, particularly in the past three decades. This review is divided into three main chapters, including microbial insecticides in pest control, utilization of semiochemicals, and botanical insecticides, paying particular attention to those practical approaches that are respectful to the environment.

Understanding the evolutionary transition from interspecific exploitation to cooperation is a major challenge in evolutionary biology. Ant-aphid relationships represent an ideal system to this end because they encompass a coevolutionary continuum of interactions ranging from mutualism to antagonism. In this study, we report an unprecedented interaction along this continuum: aggressive mimicry in aphids. We show that two morphs clonally produced by the aphid Paracletus cimiciformis during its root-dwelling phase establish relationships with ants at opposite sides of the mutualism-antagonism continuum. Although one of these morphs exhibits the conventional trophobiotic (mutualistic) relationship with ants of the genus Tetramorium, aphids of the alternative morph are transported by the ants to their brood chamber and cared for as if they were true ant larvae. Gas chromatography-mass spectrometry analyses reveal that the innate cuticular hydrocarbon profile of the mimic morph resembles the profile of ant larvae more than that of the alternative, genetically identical nonmimic morph. Furthermore, we show that, once in the brood chamber, mimic aphids suck on ant larva hemolymph. These results not only add aphids to the limited list of arthropods known to biosynthesize the cuticular chemicals of their deceived hosts to exploit their resources but describe a remarkable case of plastic aggressive mimicry. The present work adds a previously unidentified dimension to the classical textbook paradigm of aphid-ant relationships by showcasing a complex system at the evolutionary interface between cooperation and exploitation.

International audience; The sex pheromone of feral sweet potato weevils Cylas formicarius elegantulus from Cuba was found, via solid-phase microextraction analysis, to be identical to (Z)-3-dodecenyl (E)-2-butenoate, a previously reported compound. Females emitted 20 pg pheromone d−1. In scanning electron microscopy studies carried out on the male antenna, we identified several types of sensilla: sensilla trichoidea of type 1 (ST1) as long hairs (100–150 μm), sensilla trichoidea of type 2 as short hairs (50–60 μm), sensilla basiconica of type 1 as thick pegs (20–25 μm), sensilla basiconica of type 2 as curved pegs (10–15 μm), and sensilla basiconica of type 3 as thin and straight short pegs (15–20 μm). The same types were observed in female antennae but ST1 were far less abundant than in males. Sensilla chaetica were also found on the flagellum subsegments in both sexes. In electrophysiological tests, the crotonate function in the pheromone structure proved to be critical for activity since regular depolarizations (0.6–0.8 mV) were obtained with puffs on 1 μg of the attractant, but not with puffs of the formate, acetate, propionate, or butyrate analogue of the pheromone. In a double dual-choice olfactometer, males showed maximum activity between the 4th and the 8th hr of scotophase at a dose of 50–1000 ng of pheromone. In field tests, a correlation between the contents of the Z,E isomer in the pheromone formulation with activity was noticed, and baits containing this isomer of stereomeric purity >94% showed the highest attractivity. The presence of 5% of the Z,Z isomer in the lure did not induce any synergistic or inhibitory effect, and the alcohol precursor of the pheromone was inactive. The results show that use of a stereomerically pure pheromone may not be necessary in pest control strategies.

Single-cell electrophysiological recordings were obtained from olfactory receptor neurons (ORNs) in sensilla trichodea on male antennae of hybrids formed mainly by crossing female Heliothis subflexa with male Heliothis virescens (‘SV hybrids’). We recorded from the A-, B-, and C-type sensilla trichodea, with the latter two types housing ORNs exhibiting response profiles to different pheromone components that we had previously found to be characteristic for each species. For both the B- and the C-type SV hybrid sensilla, most of the ORNs exhibited a spike amplitude and ORN co-compartmentalization within sensilla that more strongly resembled the ORNs of parental H. subflexa rather than those of H. virescens. The overall mean dose-response profiles of the ORNs in hybrid C- and B-type sensilla were intermediate between those of the H. virescens and H. subflexa parental type ORNs. However, not all hybrid ORNs were intermediate in their tuning spectra, but rather ranged from those that closely resembled H. subflexa or H. virescens parental types to those that were intermediate, even on the same antenna. The most noteworthy shift in ORN responsiveness in hybrid males was an overall increase in sensitivity to Z9–14:Ald exhibited by Z9–16:Ald-responsive ORNs. Heightened cross-responsiveness to Z9–14:Ald by hybrid ORNs correlates well with observed behavioral cross-responsiveness of hybrids in which Z9–14:Ald could substitute for Z9–16:Ald in the pheromone blend, a behavior not observed in parental types. The hybrid ORN shifts involving greater sensitivity to Z9– 14:Ald also correlate well with studies of hybrid male antennal lobe interneurons that exhibited a shift toward greater cross-responsiveness to Z9–14:Ald and Z9– 16:Ald. We propose that the differences between parental H. virescens, H. subflexa, and SV hybrid male pheromone ORN responsiveness to Z9–16:Ald and Z9–14:Ald are most logically explained by an increased or decreased co-expression of two different odorant receptors for each of these compounds on the same ORN.

Toxic oil syndrome (TOS) is a disease that appeared in Spain in 1981. It affected more than 20 000 people and produced over 300 deaths in the first 2 years. In this paper, a prospective study on the differences in gene expression in sera between a control versus a TOS-affected population, both originally exposed to the toxic oil, is presented. Differential protein expression was analyzed by two-dimensional electrophoresis (2-DE). Several problems related with serum analysis by 2-DE were addressed in order to improve protein detection in the gel images. Three new commercial systems for albumin depletion were tested to optimize the detection of minor proteins that can be obscured by the presence of a few families of high abundance proteins (albumin, immunoglobulins). Other factors, such as the use of nonionic reductants or the presence of thiourea in the gels, were also tested. From these optimized images, a group of 329 major gel spots was located, matched and compared in serum samples. Thirty-five of these protein spots were found to be under- or overexpressed in TOS patients (> three-fold increase or decrease). Proteins in the differential spots were identified by matrix-assisted laser desorption/ionization-time of flight peptide map fingerprinting and database search. Several haptoglobin isoforms were found to be differentially expressed, showing expression phenotypes that could be related with TOS affection. Haptoglobin phenotypes have been previously reported to have important biological and clinical consequences and have been described as risk factors for several diseases.

5 pages, 5 figures.-- PMID: 12720381 [PubMed].-- Printed version published May 7, 2003., The sex pheromone of the oak processionary moth Thaumetopoea processionea has been characterized from female gland extracts as a mixture of (Z,Z)-11,13-hexadecadienyl acetate (1), (E,Z)-11,13-hexadecadienyl acetate (3) and (Z,Z)-11,13-hexadecadienol (2) in 88:7:5 ratio. The amount of the major compound 1 was 20−30 ng/gland. No trace of (Z,Z)-11,13-hexadecadienal was found in the extract, and therefore, T. processionea appears to be the only “summer” processionary moth lacking this compound as a pheromone compound. The alcohol 2 had also been previously found but is electrophysiologically inactive, and in wind tunnel assays it lowers the number of contacts with the source when mixed with the major compound 1. The major component 1 elicited males to display the complete behavioral sequence, but the amount of chemical needed was unexpectedly high in comparison to the activity displayed by virgin females and gland extracts. (E,E)-11,13-hexadecadienyl acetate (5) inhibits the attractant activity of the major component 1 when mixed with 1 in 1:10 and 1:1 ratios. The main constituent 1 is active in the field, but its tendency to isomerize into the corresponding E,E isomer (5) must be considered if effective formulations are to be prepared., Economic support provided by Generalitat de Catalunya and CICYT (2001SGR 00342 and projects AGF97-1217-C02-01, AGL2000-1695- C02-01 and AGL2001-0585).

A series of experiments were conducted to examine the effect of 3-octylthio-1,1,1-trifluoro-2-propanone (OTFP) on growth, development, and behavior of the cotton leafworm, Spodoptera littoralis (Lepidoptera: Noctuidae), and the corn stalk borer, Sesamia nonagrioides (Lepidoptera: Noctuidae). The chemical behaved as an oviposition deterrent and, when added to the diet of the second-instar larvae of both insects, reduced diet consumption and growth, pupation, and adult emergence. Treatment of 100-5000 ng of the compound on fourth-instar larvae for 3-24 h, however, did not produce significant differences in the amount of diet ingested. Our results suggest that the effect of OTFP is long-lasting and that the inhibitor is not fully detoxified by the detoxification enzymes of the digestive tract of the insects. In behavioral assays, adult males which had been treated with the chemical at the larval stage were less attracted to the pheromone source than regular untreated males. When Sp. littoralis untreated females were used as the attractant source, treated males also displayed significantly fewer contacts with the cage-containing females than untreated or solvent-treated males. In the presence of treated females, only 27% of treated males successfully completed the flight in comparison to animals responding to solvent-treated females (54.5%). By contrast, when Se. nonagrioides females, whether they had been subjected or not to the treatment, were used as the attractant source, males were similarly attracted to them regardless of whether they had been treated or not at the larval stage. Analyses of gland extracts of Sp. littoralis treated females showed no difference from control insects in the qualitative or quantitative composition of the pheromone blend. The results obtained, in combination with other results previously reported by us (Riba, M.; Sans, A.; Bau, P.; Grolleau, G.; Renou, M.; Guerrero, A. J. Chem. Ecol. 2001, 27, 1879-1897), provide new and relevant information about the possible utility of these chemicals in future studies directed to the development of new approaches for pest control.

Increased behavioral sensitivity to the pheromone after brief exposure of the whole insect to the sex pheromone has been documented in antennal lobe neurons of Spodoptera littoralis. We investigated whether a brief stimulus of the major component of the pheromone on naïve antenna separated from the head increased the electroantennographic responses after successive stimulations at different times. The response increase was clear 30 min after the first stimulation, and this effect lasted at least 60 min, the average life time of the antenna. Our results suggest that the olfactory receptor neurons, and not only the neurons in the antennal lobe, may be involved in the increased antennal response after a single pheromone pulse.

The flathead oak borer Coroebus undatus F. (Coleoptera: Buprestidae) is one of the primary pests of cork oak Quercus suber L. in the Mediterranean region causing great economic losses to the cork industry. Very little is known about its biology and behavior and, so far, no control measures have been established. We present the results of a pilot study aimed to develop an efficient trapping method for monitoring this harmful pest. In a 3-year field study, purple-colored prism traps baited with a mixture of green leaf volatiles (GLVs) from the host have been shown the most effective combination to catch C. undatus adults (solely females) compared to other trap and lure types tested. Wavelength and reflectance measurements revealed that purple traps exhibit reflectance peak values similar to those found in the abdominal and elytral cuticle of both sexes, suggesting the involvement of visual cues for mate location in this species. The data presented are the first to demonstrate captures of adults of the genus Coroebus by an attractant-based trapping method.

A new, convergent synthesis of (E,Z)-2,13-octadecadienyl acetate (1) and (E,Z)-3,13-octadecadienyl acetate (2), the two key components of the leopard moth Zeuzera pyrina, from 2-chloromethyltetrahydrofuran in good overall yields and stereomeric purity is reported. The synthesis of both components utilizes the common intermediate sulfone 12 as a convenient building block to be coupled with iodoacetylenic derivatives 9 or 17 in the key step.

Flight-tunnel experiments were conducted using Helicoverpa zea males to determine whether or not (Z)-11-hexadecen-1-ol (Z11-16:OH), a compound emitted by another heliothine moth species, Heliothis subflexa, is a behavioral antagonist when admixed with the two-component pheromone blend of H. zea. Males were less likely to fly upwind all the way to the source when 0.3% Z11-16:OH was present in the blend. Even 0.1% Z11-16:OH caused differences in the flight behavior of H. zea males; they steered more off the windline than males responding to the pheromone blend alone, resulting in more oblique track angles. Thus Z11-16:OH appears to act antagonistically, along with another compound, (Z)-11-hexadecen-1-ol acetate (Z11-16:Ac), when it is added to the H. zea pheromone blend.

The behavioral response of processionary males (Thaumetopoea pityocampa) to the natural pheromone (Z)-13-hexadecen-11-ynyl acetate (1) and structurally related analogs in a wind tunnel is presented. Stereomerically pureZ-1 and a mixture with theE isomer in 80:20 ratio elicited similar attraction responses at 1 µg and higher. The activity was dose-dependent, being optimum at 1 µg with 90% and 80% of males contacting with the source in the presence of theZ-1 andZ/E-1, respectively. 11-Hexadecynyl acetate (2) functioned as a pheromone mimic, being able to induce the complete mate-finding behavioral sequence, although its activity was much lower than that of the pheromone. (Z)-13-Hexadecen-11-ynyl alcohol (3) and, particularly, (Z)-13-hexadecen-11-ynal (4) were potent inhibitors of the upwind flight response in mixtures withZ-1 in 99:1, 95:5, and 91:9 ratios. (Z)-1,1,1-Trifluoro-16-nonadecen-14-yn-2-one (5) also inhibited the response of males to pheromone, particularly in the source contact behavior. Comparison with activity displayed by analogs in field tests is also reported.

The electrophysiological activity of some halogenated analogues of the major component of the sex pheromone of the corn stalk borer Sesamia nonagrioides Lef. (1) is presented. The analogues comprise a series of fluoro-, chloro- and bromoacetate analogues4–10 as well as trifluoromethyl ketone 11. The fluoro derivatives 4–6 displayed remarkable electro-antennogram (EAG) intrinsic activities in comparison with the parent acetate 1, while the remaining analogues elicited significantly lower response. The compounds have also been tested as inhibitors of the sex pheromone perception in EAG and in the field. In the laboratory. fluoro analogues 4–6 were better inhibitors than chloro derivatives 7–9, which in turn behaved similarly to the bromoacetate 10. Trifluoromethyl ketone 11, however, was a poor inhibitor of the pheromone action. In the field, baits of mixtures of compounds 5–11 with the corn stalk borer pheromone in 10:1 ratio inhibited the concomitant attraction of the clover cutworm moth Scotogramma irifolii Rott., while the difluoro analogue 5, trichloroacetate 9 and trifluoromethyl ketone 11 also diminished the number of catches of the armyworm Mythimna unipuncta Haw. The monofluoroacetate 4. trifluoro analogue 6 and bromo derivative 10 significantly disrupted the pheromone action of the corn borer, whereas trifluoromethyl ketone 11 synergistically increased the number of males attracted to the pheromone trap alone. Addition of 11 to baits containing the corn borer pheromone caught S. nonagrioides selectively with regard to the other habitat-sharing species M. unipuncta and S. trifolii.

Some twig beetles in the genus Pityophthorus (Coleoptera: Scolytinae) may vector pitch canker disease Fusarium circinatum (NirembergO'Donnell) of Pinus spp. (Pinaceae). Because Pityophthorus pubescens (Marsh.) has been found to be associated with F. circinatum in the Basque Country (northern Spain), various experiments were conducted to assess the beetle's behavioural responses to (E, E)-α-farnesene, (R)-(+)-limonene and (S)-(-)-verbenone to develop a potential inhibitor to host attraction. These experiments comprise electroantennographic and double-choice olfactometer tests, as well as field assays in Pinus radiata D. Don stands.Both sexes of P. pubescens showed similar electroantennographic responses to different doses (from 1 ng to 1 µg in decadic steps) of each individual compound, with depolarisations to (S)-(-)-verbenone (100 ng) being similar to those of the aggregation pheromone (+)-trans-pityol. In olfactometer assays, both sexes were significantly attracted to (+)-trans-pityol, but the attraction was reduced when increasing amounts of the chemicals were added to the pheromone. Particularly relevant was the repellent effect induced by (S)-(-)-verbenone at 1 ng dose and higher. In the field, (E, E)-α-farnesene, (R)-(+)-limonene and (S)-(-)-verbenone reduced significantly the number of beetles attracted to (+)-trans-pityol and racemic trans-pityol, with (S)-(-)-verbenone being the most effective.(S)-(-)-Verbenone showed an interesting potential for use in the protection of P. radiata stands. A potentially effective strategy, which could be implemented in further, more in-depth studies, could involve the use of this semiochemical as repellent and (+)-trans-pityol-baited traps as attractant in a 'push-pull' strategy.

The inner bark of Ceylon cinnamon (Cinnamomum zeylanicum L.) is commonly used as a spice and has also been widely employed in the treatment and prevention of disease. The positive health effects associated with the consumption of cinnamon could in part be due to its phenolic composition; proanthocyanidins (PA) are the major polyphenolic component in commercial cinnamon. We present a thorough study of the PA profile of cinnamon obtained using matrix-assisted laser desorption/ionization tandem time-of-flight (MALDI-TOF/TOF) mass spectrometry. In addition to the advantages of MALDI-TOF as a sensitive technique for the analysis of high-molecular-weight compounds, the tandem arrangement allows the identification of the compounds through their fragmentation patterns from MS/MS experiments. This is the first time that this technique has been used to analyze polymeric PA. The results show that cinnamon PA are more complex than was previously thought. We show here for the first time that they contain (epi)gallocatechin and (epi)catechingallate units. As gallates (galloyl moieties) and the pyrogallol group in gallocatechins have been related to the biological activity of grape and tea polyphenols, the presence of these substructures may explain some of the properties of cinnamon extracts. MALDI-TOF/TOF reveals that cinnamon bark PA include combinations of (epi)catechin, (epi)catechingallate, (epi)gallocatechin, and (epi)afzelechin, which results in a highly heterogeneous mixture of procyanidins, prodelphinidins, and propelargonidins., This work was supported by the Spanish Ministry of Education and Science (research grants AGL2009-12374-C03-03/ALI). J.P-J thanks the Spanish Ministry of Science and Innovation for granting her a Sara Borrell postdoctoral contract (CD09/00068).

Objectives: Toxic oil syndrome (TOS) is a disease that appeared in Spain in 1981. Epidemiological work traced the origin to the ingestion of aniline-adulterated rapeseed oil, fraudulently marketed and sold as edible oil. It affected more than 20,000 people with over 400 deaths in the first 2 years. In 2001 evidence was presented that genetic factors could play a role in the susceptibility of individuals to the disease. Thus, a prospective study on the differences in gene expression in sera between control versus TOS-affected populations, both originally exposed to the toxic oil, was undertaken in our laboratory. Methods: Differential protein expression was analyzed by two-dimensional electrophoresis (2-DE). Problems related with serum analysis by 2-DE were addressed to improve protein detection in the gel images. Three new commercial systems for albumin depletion were tested to optimize the detection of minor proteins. The use of nonionic reductants or the presence of thiourea in the gels, were also tested. Results: From the resulting optimized images, a group of 329 major gel spots was located, matched and compared with serum samples. Thirty-five of these protein spots were found to be under- or over-expressed in TOS patients (threefold increase or decrease). Proteins in these spots were identified by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) peptide map fingerprinting and database search. Several haptoglobin (Hp) isoforms were found to be differentially expressed, showing expression phenotypes that could be related with TOS. Resolution of the homologous α-1s and α-1f chains, with a mass difference of only 0.043 Da, was obtained after guanidation of the protein with O-methylisourea. We applied this procedure to the study of the distribution of the Hp alleles HP 2, HP 1s and HP 1f in control versus TOS-affected populations. The MALDI-TOF proteotyping method was validated by a parallel analysis of the serum samples by 2-DE. Conclusions: Data obtained from 54 TOS cases and 48 controls indicate significant differences in the distribution of Hp phenotypes in the two populations. Haptoglobin phenotypes have been reported to have biological and clinical consequences and have been described as risk factors for several diseases. Consequently, it was concluded that haptoglobin polymorphism could play a role in TOS. © 2010 Elsevier Ireland Ltd. All Rights Reserved., This work was supported by WHO projects EU/03/016989 and EU 05/025039.

Pest damage causes important decrease in crop yield every year all over the world, particularly by Lepidoptera. Characterization of the antennal proteins implicated in the reproduction of Lepidoptera will help to develop new methods for pest management and contribute to sustainable agriculture and biodiversity maintenance. We present herein the characterization of some antennal proteins of Sesamia nonagrioides by proteomic techniques such as two-dimensional electrophoresis, MALDI-TOF MS, and electrospray ionization tandem mass spectrometry (ESI-MS/MS). The antennal proteins expressed in both sexes were analyzed and more than 800 spots were detected, finding 16 proteins differentially expressed between males and females. Most of the identified proteins were involved in olfaction. High levels of pheromone binding proteins (PBP1 and PBP2) were found as expected in males, but also in female antennae, although females did not electrophysiologically respond to their own pheromone. General odorant binding proteins (GOBP1 and GOBP2) were preferentially expressed in females but high levels were also detected in males. The expression was remarkably high in both sexes along the complete photoperiod. A sensitive proteomic methodology was developed to identify antennal proteins., We gratefully acknowledge CSIC for an I3P fellowship to P. Acín and to MEC for a Ramon y Cajal Contract to C. Quero. Financial support was obtained from CICYT (project AGL2006-13489-C02-01).

10 pages, 6 figures, 3 tables.-- PMID: 16544284 [PubMed]., Toxic oil syndrome (TOS) is a disease that appeared in Spain in 1981 as a consequence of the ingestion of an aniline-adulterated oil illegally marketed as edible. TOS affected more than 20 000 people and produced over 400 deaths in the first 18 months after the outbreak. There is evidence that genetic factors could play a role in the susceptibility of individuals towards the disease. Recently, we suggested that haptoglobin (Hp) polymorphism could also play a role in TOS. To provide a rapid method for high-throughput Hp phenotyping, we developed a two-step MALDI-TOF procedure that allows specific identification of the three common Hp chains. Resolution of the homologous α-1s and α-1f chains, which have a mass difference of only 0.043 Da, is obtained after guanidination of the protein with O-methylisourea. We applied this procedure to the study of the distribution of the Hp alleles HP(1s), HP(1f), HP2 in a control versus a TOS-affected population, both originally exposed to the toxic oil. The MALDI-TOF proteotyping method was validated by a parallel analysis of the serum samples by 2-DE. Data obtained from 54 TOS cases and 48 control individuals indicate significant differences in the distribution of Hp phenotypes in the two populations., This work was supported by WHO projects EU/03/016989 and EU 05/025039.

Single-cell electrophysiological recordings were obtained from olfactory receptor neurons in sensilla trichodea on male antennae of the heliothine species Heliothis subflexa and the closely related congener H. virescens. A large percentage of sensilla (72% and 81%, respectively, of all sensilla sampled) contained a single odor-responsive receptor neuron tuned to the major pheromone component of both species, Z-11-hexadecenal. A second population of sensilla on H. subflexa antennae (18%) housed receptor neurons that were tuned to Z-9-hexadecenal but also responded with less sensitivity to Z-9-tetradecenal. A similar population of sensilla (4%) on H. virescens male antennae housed receptor neurons that were shown to be tuned specifically only to Z-9-tetradecenal, with no response to even high dosages of Z-9-hexadecenal. A third population of sensilla (comprising 8% and 16% of the sensilla sampled in H. subflexa and H. virescens, respectively) housed two olfactory receptor neurons, one of which was tuned to Z-11-hexadecenyl acetate and the other tuned to Z-11-hexadecenol. In H. subflexa the Z-11-hexadecenyl acetate-tuned neuron also responded to Z-9-tetradecenal with nearly equivalent sensitivity. The behavioral requirements of males of these two species for distinct pheromonal blends was, therefore, reflected by the subtle differences in the tuning properties of antennal olfactory receptor neurons.

Increased behavioral sensitivity to the pheromone after brief exposure of the whole insect to the sex pheromone has been documented in antennal lobe neurons of Spodoptera littoralis. We investigated whether a brief stimulus of the major component of the pheromone on naïve antenna separated from the head increased the electroantennographic responses after successive stimulations at different times. The response increase was clear 30 min after the first stimulation, and this effect lasted at least 60 min, the average life time of the antenna. Our results suggest that the olfactory receptor neurons, and not only the neurons in the antennal lobe, may be involved in the increased antennal response after a single pheromone pulse.

Butterflies and moths are subject to different evolutionary pressures that affect several aspects of their behaviour and physiology, particularly sexual communication. Butterflies are day-flying insects (excluding hedylids) whose partner-finding strategy is mainly based on visual cues and female butterflies having apparently lost the typical sex pheromone glands. Moths, in contrast, are mostly night-flyers and use female-released long-range pheromones for partner-finding. However, some moth families are exclusively day-flyers, and therefore subject to evolutionary pressures similar to those endured by butterflies. Among them, the Castniidae, also called ‘butterfly-moths’ or ‘sun-moths’, behave like butterflies and, thus, castniid females appear to have also lost their pheromone glands, an unparallel attribute in the world of moths. In this paper, we review the sexual communication strategy in day-flying Lepidoptera, mainly butterflies (superfamily Papilionoidea), Zygaenidae and Castniidae moths, and compare their mating behaviour with that of moth families of nocturnal habits, paying particular attention to the recently discovered butterfly-like partner-finding strategy of castniids and the fascinating facts and debates that led to its discovery.

Increased behavioral sensitivity to the pheromone after brief exposure of the whole insect to the sex pheromone has been documented in antennal lobe neurons of Spodoptera littoralis. We investigated whether a brief stimulus of the major component of the pheromone on naïve antenna separated from the head increased the electroantennographic responses after successive stimulations at different times. The response increase was clear 30 min after the first stimulation, and this effect lasted at least 60 min, the average life time of the antenna. Our results suggest that the olfactory receptor neurons, and not only the neurons in the antennal lobe, may be involved in the increased antennal response after a single pheromone pulse.