EXPLOITING PHEROMONES AND RELATED CHEMICALS FOR DETECTION, MONITORING, AND CONTROL OF INSECT PESTS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 1002931
• Project Start Date: Sep 1, 2014
• Project End Date: Aug 31, 2019
• Program Code: [(N/A)]- (N/A)

Project Director
Millar, JO.

Recipient Organization
UNIVERSITY OF CALIFORNIA, RIVERSIDE
(N/A)
RIVERSIDE,CA 92521

Performing Department
Entomology, Riverside

Non Technical Summary
This project will establish and coordinate multidisciplinary efforts to define and solve problems in insect pest management, with specific emphasis on developing, validating, and delivering new tools and strategies for the management of insect pests. The major focus will be on the chemistry of bioactive compounds such as pheromones that insect rely on for communication with each other, and the development of methods for exploiting these chemicals. Particular emphasis will be placed on invasive species, to minimize their impacts on Californian and US agriculture, forests, and natural environments. Because this work is multidisciplinary by nature, it will involve numerous collaborators, such as other members of the department, Cooperative Extension personnel, grower groups, USDA or other regulatory personnel, and others with specialized expertise as needed to provide the critical mass of expertise and resources required to carry out particular projects. The mandate of the parts of this project have been left as broad as possible so that subprojects involving essentially any insect species or commodity can be accomodated, so as to be able to mount rapid responses to the continuing influx of new pest threats. Within this mandate, projects will be conducted on a rolling basis, with new projects being introduced as others are finished. In the near term, we will focus on three projects currently being initiated or in progress. These are:1) identification and exploitation of the chemical signals used by a large family of wood-boring insects (the Cerambycidae), many of which are invasive, tree-killing species.2) "proactive chemical ecology", loosely defined as the identification of pheromones and related chemical signals that can be exploited for detection and control of invasive insect pests that are not yet present or not yet firmly established in the US;3) A detailed examination of the roles of contact pheromone signals as mediators of insect behavior, particularly for insects that live in large groups such as termites, ants, wasps, and bees, and manipulation of these signals for insect management.

Animal Health Component

50%

Research Effort Categories

Basic

50%

Applied

50%

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
211	0699	1130	75%
211	1119	2000	25%

Knowledge Area
211 - Insects, Mites, and Other Arthropods Affecting Plants;

Subject Of Investigation
1119 - Deciduous tree fruits, general/other; 0699 - Trees, forests, and forest products, general;

Field Of Science
1130 - Entomology and acarology; 2000 - Chemistry;

Keywords

pheromone

invasive species

semiochemical

detection

monitoring

cerambycid

Goals / Objectives
Goal: To establish and coordinate multidisciplinary efforts to define and solve problems in insect pest management, with specific emphasis on developing, validating, and delivering new tools and strategies for the management of insect pests.Objectives:To identify, synthesize, and develop semiochemicals used by native and exotic cerambycid beetles.To proactively identify semiochemicals of other exotic pests that are likely to be introduced into the US through global commerce, and develop lures that can be used for their detection.To explore the role of contact pheromones as mediators of insect behavior, and where feasible, develop methods for exploiting those signals for insect management.To conduct preliminary studies on newly discovered invasive species, both to get a head start on developing management programs, and to obtain preliminary data to drive grant proposals.

Project Methods
Objective 1. To identify, synthesize, and develop semiochemicals used by native and exotic cerambycid beetles.Three strategies will be used. First, we and others have shown that cerambycid pheromone structures are generally highly conserved, with species within the same genus, tribe, or even subfamily using the same or very similar pheromone components (e.g., Hanks and Millar 2013), even between different continents (e.g., Wickham et al. 2014; Imrei et al. 2013). We have synthesized a library of the known cerambycid pheromones, and with the help of collaborators, we will continue screening these pheromones in new areas. For example, we are initiating new collaborations in central Brazil and Kenya this year. Overall, this is proving to be a fast and very cost-effective method of identifying likely pheromones for exotic species, and it has already resulted in the identifications of pheromones for several potentially dangerous invaders (e.g. Teale et al. 2011; Pajares et al. 2014).Second, specific exotic species deemed of high risk for invasion by a USDA-APHIS risk assessment are being targeted for pheromone identification. These exotic species will be accessed through our network of collaborators, with my group providing the chemistry support, i.e., we will assist with or fully carry out the identification of new semiochemicals from samples or spectra collected by collaborators, and synthesize the compounds for field testing by the collaborators. We have already demonstrated the efficacy of this approach by identifying pheromones from European (Larssen et al. manuscript in prep.) and Asian species (Teale et al. 2011), and we are currently working with collaborators in China on the identifications of several other important species (e.g. Anaplophora chinensis and Massicus raddei).Third, because cerambycid pheromone structures are generally highly conserved within taxonomic groups, even between continents, for exotic species which are difficult to obtain, we will use an "identification by proxy" strategy. That is, important Asian species such as Semanotus bifasciatus have North American congeners such as Semanotus litigiosus, which is a common species in southern California. Thus, identifying the pheromone of the latter species should inform us as to what type(s) of compounds are likely to constitute the pheromone of the dangerous Asian species. All the apparatus and equipment required for the collection, identification and synthesis of cerambycid pheromones is in place, including aeration chambers, coupled GC-electroantennographs, GC-mass spectrometers, and a fully equipped synthesis laboratory. My group also has a decade of experience in working with cerambycid pheromones.Objective 2. To proactively identify semiochemicals of other exotic pests that are likely to be introduced into the US through global commerce, and develop lures that can be used for their detection. Work under this objective will take place in several sequential steps:Identification of exotic pests that represent a high risk to California/US agriculture, forests, or landscapes. Target pests will be selected in consultation with a variety of sources, including regulatory agencies, commodity groups, and international collaborators.Logistical planning and execution of work. For a particular exotic target insect, wherever possible we will establish collaborations with colleagues in the native country of the insect, to minimize expenses. The in-country colleagues will be responsible for initial collections of the insects, and the testing of lures once compounds have been identified and synthesized. We will be responsible for identification of the new semiochemicals, using extracts prepared and shipped by collaborators to UCR. My group will also synthesize the compounds. To assist with this work, we are finishing up building a portable gas chromatograph-electroantennogram detector. It can be taken to any desired country to do conduct the key preliminary analyses of pheromone extracts (i.e., location of the bioactive compounds in the extract). Once we have fixed the locations of the compounds in a GC trace with reference to standards, we can then reliably relocate the compounds in larger extracts shipped back to UCR for full identification of the compounds. This instrument will circumvent a number of the bottlenecks that have hindered our efforts to date to identify semiochemicals of exotic species.Objective 3. To explore the role of contact pheromones as mediators of insect behavior, and where feasible, develop methods for exploiting those signals for insect management. Most of our work under this objective will be collaborative, with my group doing the analysis of extracts, and the isolation, identification, and synthesis of bioactive compounds to verify their activities, and to explore the possibilities for using them for insect management. This multidisciplinary and multigroup approach should be highly synergistic. The basic protocol consists of a series of steps in which we fractionate cuticular lipids into different classes, and then separate the members of each group by molecular size and degree of branching. The steps consist of:1. Separation of crude extracts into saturated alkanes, unsaturated hydrocarbons, and more polar compounds.2. Separation of saturated alkanes into straight-chain and branched chain fractions using molecular sieves.3. Separation of unsaturated hydrocarbons into mono-, di-, and triunsaturated compounds, and more highly unsaturated compounds.4. Separation of subfractions from steps 2 and 3 on the basis of chain length and branch points by reverse phase HPLC, and detection by evaporative light scattering detection. The latter is critical, because saturated and to some extent unsaturated hydrocarbons are invisible to most HPLC and LCMS detectors.5. Determination of chirality of isolated branch chain compounds by polarimetry. We have also been working with an HPLC detector company to determine whether it might be possible to determine chirality on the fly using a polarimeter optimized for use as an HPLC detector.6. Determination of double bond positions and geometries in unsaturated compounds using a combination of microscale derivatization and fragmentation.7. As needed, synthesis of pure standards for confirmation of structures, and laboratory and field bioassays to be conducted by collaborators. We currently have collaborations planned or in progress to work with cuticular lipids of Drosophila (several research groups), Argentine and trapjaw ants, vespid and parasitic wasps, and several species of bees. All the equipment required for this work is in place, as is the expertise to isolate, identify, and synthesize any desired insect cuticular lipid.Objective 4. To conduct rapid initial studies on newly discovered invasive species, both to get a head start on developing management programs, and to obtain preliminary data to help drive grant proposals. We are fully equipped to conduct rapid preliminary studies to determine whether new invasive species are likely to use semiochemicals that could be exploited in containment and eradication programs, and to identify and synthesize those compounds. We have several decades of expertise with these types of compounds, in conducting preliminary bioassays with them. Whereas it is impossible to predict which exotic species will invade and when, it is guaranteed that some will arrive and establish. It is important for my research group to maintain the capability to respond rapidly to new pest threats as they emerge.

Progress 10/01/18 to 08/31/19

Outputs
Target Audience:Audiences targeted and reached by this project included: 1. Academics and research scientists working in chemical ecology and entomology, with an emphasis of applying chemical ecology to protection of crops and natural habitats. 2. Regulatory officials working in local, state, national, and international agencies charged with crop protection and with detection and management of invasive species. 3. Agricultural industry/commodity boards and related grower groups via talks at commodity group meetings, and one on one discussions with commodity board officials and grower representatives. 4. The general public through outreach events such as the annual UCR Insect Fair held in downtown Riverside. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Opportunities for training and professional development included: 1. Ongoing training and mentoring of two PhD students in Entomology, one of whom is a Hispanic woman (Jacqueline Serrano) who is the first in her family to go to college, and one PhD student in Chemistry. Ms Serrano successfully graduated in August 2019. 2. Sending students to regional, national conferences, and international conferences, where they gave presentations. Conferences included the annual meetings of the Pacific branch of the Entomological Society of America, the joint meeting of the Entomological Societies of America and Canada, and the annual meeting of the International Society for Chemical Ecology. How have the results been disseminated to communities of interest?Results have been disseminated to communities of interest through: 1. Sixteen papers published in mainstream refereed scientific publications. 2. Presentations at regional, national, and international conferences, and commodity group meetings (California Almond Board and California Pistachio Commission). During the report period, the PI was author or coauthor on 7 invited and 12 submitted presentations. 3. To expedite technology transfer and uptake of our research into pest surveilance and management programs, Immediately after acceptance, relevant publications have been sent to contacts in pheromone products companies, including Trece, IscaTech, Chemtica, Synergy, and Alpha Scents. What do you plan to do during the next reporting period to accomplish the goals?Work planned for the next reporting period, by objective: Objective 1. To identify, synthesize, and develop semiochemicals used by native and exotic cerambycid beetles. Over the winter, we have been working to identify and synthesize additional new compounds, to have them ready for field testing this coming season in the US, and by project collaborators in Asia and Europe. Field trials will extend through most of the coming year because from September onwards, we will be working with our collaborators in the southern hemisphere. We also have extracts from additional species, which contain as yet unidentified compounds, and so as quantities permit, these compounds will be isolated for microbore NMR spectrometry to provide sufficient information to fully identify them. As an indication of how productive this work has been and continues to be, since 2004, we and our network of collaborators have identified pheromones or likely pheromones for several hundred species from all continents except Antarctica, including for a number of invasive species which are already in the US, and other species which are deemed at high risk of invading through international commerce in wood and wood products. Development of these pheromones is providing effective new tools for US regulatory agencies to detect exotic species if they do invade, and help in the eradication of localized populations before they have a chance to spread. A number of pheromone products companies are now carrying cerambycid pheromone product lines, in large part as a result of our research. Objective 2.To proactively identify semiochemicals of other exotic pests that are likely to be introduced into the US through global commerce, and develop lures that can be used for their detection. Fueled by our recent successes in identifying and bioassaying the first pheromones for North American click beetle species, including the major pest Melanotus communis, we are expanding our studies to include species from a number of different genera, including Limonius, additional Melanotus spp., Dalopius, and Conoderus. Although these insects are native, they are of increasing importance as crop pests, particularly in grains, corn, and root vegetables, in part because the new generation of insecticides is proving to be relatively ineffective against these insects. This project will be carried forward with collaborators in Washington, Idaho, Illinois, North and South Carolina, and Canada. Objective 3: To explore the role of contact pheromones as mediators of insect behavior, and where feasible, develop methods for exploiting those signals for insect management. We have ongoing collaborations with groups at UC Riverside, University of Arizona, North Carolina State University, and University of Georgia, as well as several groups overseas, studying the roles of queen pheromones and other cuticular lipids in fire ants, trap-jaw ants, yellowjacket wasps, honeybees, and bumble bees. Our role in these studies is to isolate, identify, and synthesize the bioactive compounds, so that the full details of the required components of the active pheromones, and the functional roles of specific individual compounds (as opposed to crude extracts or fractions of extracts of variable composition), can be worked out. This in turn may provide leads for developing methods of controlling pest social insect species, and particularly invasive species, by manipulation or disruption of the queen signals, or signals indicating nestmate status or caste. Objective 4.To conduct preliminary studies on newly discovered invasive species, both to get a head start on developing management programs, and to obtain preliminary data to drive grant proposals. We have initiated a collaboration with researchers in New Zealand to study pheromones of invasive wasp species in the genus Vespula, focusing on North American pest species that have invaded New Zealand and Hawaii. Because these species (e.g., Vespula germanica and Vespula pensylvanica) are pests in both native and invaded ranges, anything that we discover should be widely applicable for their management and control worldwide. To date, in analyses of extracts of V. pensylvanica, we have identified and synthesized novel compounds from both virgin queens and drones. It is virtually certain that these compounds will be involved in reproductive interactions in these species.

Impacts
What was accomplished under these goals? Accomplishments by objective: Objective 1. To identify, synthesize, and develop semiochemicals used by native and exotic cerambycid beetles. During the report period, several new pheromones were identified for species in the genera Eburodacrys, Compsibidion, Eutrypanus, Oncideres, and Epacroplon. Most of these compounds have been synthesized, with the remainder in progress. The synthesized compounds are undergoing field testing in the US, and with collaborators, on other continents. In particular, because our previous research has shown that pheromone structures are often highly conserved among related cerambycid species, we have repeatedly shown that a pheromone component identified from e.g., a North American species, may also be a pheromone component of related species on other continents, and field screening of newly identified pheromones has greatly accelerated the identifications of pheromones for additional species worldwide, including a number of invasive species. For example, we have shown that the common cerambycid pheromone component 3-hydroxyhexan-2-one is a pheromone component for cerambycid species from all six habitable continents. Another important development has been our determination, now with a number of species from different continents, that the chirality of cerambycid pheromones can be crucially important to their activity. We have now shown that: a) For those species producing pheromone components that can exist in two enantiomeric forms, most but not all species produce only one of the two enantiomers. b) Some species produce both enantiomers of a given pheromone component, and they only respond to the correct blend of those two enantiomers, i.e., synergism between enantiomers. c) Some species produce only one enantiomer of a given pheromone component, but their responses are strongly antagonized by the presence of the other enantiomer. Thus, for these species, the cheaper racemic mixture of the two enantiomers will not work, and to be trapped effectively, lures will have to contain only the correct enantiomer. Overall, this has important implications for trapping protocols, and how the identified pheromones can be used both most effectively, and most economically. During the report period, we published the results of the identifications and field trials of cerambycid species in the genera Sussuacanga, Paranoplium, Plagionotus, Astylopsis, Leptostylus, Anaglyptus, Xylotrechus, and Psapharochrus, and the pheromones of the known invasive species Arhopalus rusticus, and Aromia bungii. The latter species is a notorious pest of stone fruits which has invaded Japan and parts of Europe. USDA-APHIS has categorized this species as very high risk for the US. Identification and optimization of this and the other pheromones described above will provide regulatory agencies with powerful tools for detections of new incursions, and for helping in eradication efforts if populations of exotic cerambycids do become established in the US. Objective 2. To proactively identify semiochemicals of other exotic pests that are likely to be introduced into the US through global commerce, and develop lures that can be used for their detection. In 2015, we initiated a study on the sex attractant pheromones of North American wireworm species. Although these species are endemic rather than invasive, they are rapidly becoming significant problems in many crop systems due to changes in insecticide use. No pheromones were known from any North American species; two reports of pheromone identifications from several decades ago have proven to be incorrect. In 2016, we obtained the first major positive results from this study, with pheromones or likely pheromones being identified from 5 species in different genera. In 2017, we synthesized and field tested these pheromones, verifying their activity. The importance of these results cannot be overemphasized: these represent the first correct identifications of pheromones for any of the >1,000 species of North American wireworm species, some of which are major agricultural pests. During the report period, and working with USDA and university-based collaborators in Washington, Idaho, Illinois, South Carolina, and British Columbia, we identified likely pheromones from species in the genera Melanotus, Elator, Dalopius, and Parallelostethus, and we published the pheromone of the corn wireworm, Melanotus communis, an important pest of potato and other root crops in the eastern US. Field trials with these compounds and a series of analogs will be carried out this coming field season. Overall, this project is rapidly picking up interest and momentum, and our collaborators will be collecting and shipping additional target species to us for pheromone identifications this coming field season. The appropriate permit allowing shipment of native wireworm adults into the UCR quarantine facility is in place. During the report period, we also published a first paper on the isolation of a powerful attractant for the invasive bagrada bug, Bagrada hilaris, from cauliflower seedlings. We have now identified this novel compound, given the common name brassicadiene, as a tricyclic sesquiterpene hydrocarbon, and the manuscript describing its identification has been submitted for publication. Objective 3: To explore the role of contact pheromones as mediators of insect behavior, and where feasible, develop methods for exploiting those signals for insect management. My group is collaborating with several different groups on various aspects of contact pheromones. First, we are working with Prof. Ken Ross in Georgia, with the goal of determining the chemical signals that distinguish monogyne (single queen) from polygyne (multiple queens) colonies of fire ants, a major pest across the southern US, and which invaded California about 15 years ago. Second, we are continuing a collaboration with Olav Rueppell at NC State on a project to identify CHC markers that identify honeybees as being diseased. Third, we have initiated a collaboration with Etya Ansalem at Penn State to study contact semiochemicals in bumblebee pollinators. The overall goal of these and other ongoing collaborations with entomologists studying social insects is to gain a better understanding of how these chemical signaling systems work in controlling the organization of colonies which can total millions of individuals. Our joint studies may identify signals or behaviors which can be exploited to disrupt colony organization, resulting in partial or complete control of species which are pests, without having to resort to insecticides. During the report period, we published a paper on contact pheromones in primitively eusocial wasps with collaborator Tom Wenseleers. Objective 4.To conduct preliminary studies on newly discovered invasive species, both to get a head start on developing management programs, and to obtain preliminary data to drive grant proposals. Working with Prof. Rory McDonnell at Oregon State University and Amy Roda, USDA-ARS in Florida, we developed an essential oil-based attractant for the highly invasive Giant African land snail, Lissachatina fulica, which has now been published. We have also discovered a more generic and more powerful attractant for this invasive species, and a number of other invasive gastropod species, including Cornu aspersum, and the invasive slug, Deroceras reticulatum. This work is currently being written up for publication.

Publications

• Type: Journal Articles Status: Published Year Published: 2018 Citation: Salvatore Guarino, Mokhtar Abdulsattar Arif, Jocelyn G. Millar, Stefano Colazza, Ezio Peri. 2018. Novel diterpenes emitted by seedlings of Brassica species provide host location cues to Bagrada hilaris. PLoS One. 13(12): e0209870. https://doi.org/10.1371/journal.pone.0209870
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 341. Zolt�n Imrei, Mikael A. Molander, Inis B. Winde, Zs�fia Lohonyai, �va B�lintn� Csonka, J�zsef Fail, Lawrence M. Hanks, Yunfan Zou, Jocelyn G. Millar, Mikl�s T�th, Mattias C. Larsson. 2019. Identification of the aggregation-sex pheromone of Plagionotus arcuatus ssp. arcuatus (Coleoptera: Cerambycidae) from two geographically separated populations. Naturwissenschaften 106, 18:9. https://doi.org/10.1007/s00114-019-1612-2
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 342. Yunfan Zou, Laurel Hansen, Tian Xu, Steven A. Teale, Dong Hao, and Jocelyn G. Millar. Optimizing pheromone-based lures for the invasive red-necked longhorn beetle, Aromia bungii Journal of Pest Science 92: 1217-1225. https://doi.org/10.1007/s10340-019-01108-6
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 343. Robert Maxwell Collignon; Sean Halloran; Jacqueline M. Serrano; J. Steven McElfresh; Jocelyn G. Millar. 2019. An Unstable Monoterpene Alcohol as a Pheromone Component of the Longhorned Beetle Paranoplium gracile (Coleoptera: Cerambycidae)" Journal of Chemical Ecology 45:339-347. https://doi.org/10.1007/s10886-019-01063-7
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 348. Robert M. Collignon; Jonathan A. Cale; Jocelyn G. Millar. 2019. Dose and density effects on the use of aggregation-sex pheromones by longhorn beetles (Coleoptera: Cerambycidae), and their possible role in optimal density strategy. Journal of Chemical Ecology 45: 217-226
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 344. Cintia Akemi Oi, Ricardo Caliari Oliveira, Jelle S. van Zweden, Sidnei Mateus, Jocelyn G. Millar, Fabio S. Nascimento, Tom Wenseleers. 2019. Do primitively eusocial wasps use queen pheromones to regulate reproduction? A case study of the paper wasp Polistes satan. Frontiers in Ecology and Evolution, Front. Ecol. Evol., 11 June 2019| https://doi.org/10.3389/fevo.2019.00199
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 345. Linnea R. Meier " Jocelyn G. Millar " Judith A. Mongold-Diers " Lawrence M. Hanks. 2019. (S)-Sulcatol is a Pheromone Component for Two Species of Cerambycid Beetles in the Subfamily Lamiinae. Journal of Chemical Ecology 45: 447-454
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 347. Mikael A. Molander1* " Bj�rn Eriksson1 " Inis B. Winde1 " Yunfan Zou2 " Jocelyn G. Millar2 " Mattias C. Larsson. 2019. The Aggregation-sex Pheromones of the Cerambycid Beetles Anaglyptus mysticus and Xylotrechus antilope ssp. antilope: New Model Species for Insect Conservation through Pheromone-based Monitoring. Chemoecology 29: 111-124
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 349. Jacqueline M. Serrano, J. Steven McElfresh, Yunfan Zou, and Jocelyn G. Millar. 2019. Identification of Aggregation-Sex Pheromone Components for a Living Fossil, the False Click Beetle, Palaeoxenus dohrni Horn (Coleoptera: Eucnemidae). Journal of Chemical Ecology 45: 366-370. doi.org/10.1007/s10886-019-01068-2
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 350. Ann M. Ray, Joseph A. Francese, Yunfan Zou, Kristopher Watson, Damon J. Crook, Jocelyn G. Millar. 2019. Isolation and identification of a male-produced aggregation-sex pheromone for the velvet longhorned beetle, Trichoferus campestris (Faldermann). Scientific Reports, 9:4459 | https://doi.org/10.1038/s41598-019-41047-x
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 352. Mikael A. Molander, Inis B. Winde, Joseph Burman, Franklin N. Nyabuga, Tobias U. T. Lindblom, Lawrence M. Hanks, Jocelyn G. Millar, Mattias C. Larsson. 2019. Common cerambycid pheromone components as attractants for longhorn beetles (Cerambycidae) breeding in ephemeral oak substrates in northern Europe. Journal of Chemical Ecology 45: 537-548
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 353. Hanks LM, Mongold-Diers JA, Mitchell RF, Zou Y, Wong JCH, Meier LR, Johnson TD, Millar JG. 2019. The role of minor pheromone components in segregating 14 species of cerambycid beetles of the subfamily Cerambycinae. Journal of Economic Entomology 112: 2236-2252
• Type: Journal Articles Status: Published Year Published: 2018 Citation: 333. Weliton D. Silva, Jos� Maur�cio S. Bento, Lawrence M. Hanks, Jocelyn G. Millar. 2018. (Z)-7-Hexadecene is an aggregation-sex pheromone produced by males of the South American cerambycid beetle Susuacanga octoguttata. J. Chemical Ecology 44: 1115-1119
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 354. Amy Roda; Jocelyn G. Millar; Christopher Jacobsen; Robin Veasey; Lenny Fujimoto; Arnold Hara; Rory McDonnell. 2019. A new synthetic lure for management of the invasive giant African snail, Lissachatina fulica. PLoS One 14(10):e0224270.https://doi.org/10.1371/journal.pone.0224270
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 357. Alenka }uni?-Kosi,Nataaa Stritih-Peljhan, Yunfan Zou, J. Steven McElfresh, and Jocelyn G. Millar A male-produced aggregation-sex pheromone of the beetle Arhopalus rusticus (Coleoptera: Cerambycidae, Spondylinae) may be useful in managing this invasive species. Scientific Reports 9:19570 | https://doi.org/10.1038/s41598-019-56094-7
• Type: Journal Articles Status: Published Year Published: 2018 Citation: 331. Kevin J. Loope, Jocelyn G. Millar, Erin Wilson Rankin. 2018. Weak nestmate discrimination behavior in native and invasive populations of a yellowjacket wasp (Vespula pensylvanica). Biological Invasions pp 1-14. https://doi.org/10.1007/s10530-018-1783-3

Progress 09/01/14 to 08/31/19

Outputs
Target Audience:Audiences targeted and reached by this project included: 1. Academics and research scientists working in chemical ecology and entomology, with an emphasis of applying chemical ecology to protection of crops and natural habitats. 2. Regulatory officials working in local, state, national, and international agencies charged with crop protection and with detection and management of invasive species. 3. Agricultural industry/commodity boards and related grower groups via talks at commodity group meetings, and one on one discussions with commodity board officials and grower representatives. 4. The general public through e.g., the annual Entomology department sponsored Insect Fair in downtown Riverside, and similar outreach events. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Opportunities for training and professional development included: 1. Training and mentoring of three PhD students in Entomology, and two PhD students in Chemistry. Three of these students have now graduated, two of whom have postdoctoral positions in USDA-ARS laboratories, and the third is working for a start-up company which is developing enzyme-based methods of producing insect pheromones. 2. Training and mentoring of two postdoctoral scholars. 3. Training and mentoring visiting scientists from Brazil, Slovenia, and China. This has been particularly useful because all three of these scientists are continuing to collaborate with us, in particular by field testing of new pheromones in their home countries. 4. Sending students to regional, national conferences, and international conferences, where they gave presentations. Conferences included the annual meetings of the Pacific branch of the Entomological Society of America, annual meetings of the Entomological Society of America, and annual meetings of the International Society for Chemical Ecology. How have the results been disseminated to communities of interest?During the project period, results were disseminated to communities of interest through: 1. More than 50 papers published in mainstream refereed scientific publications. 2. Presentations at regional, national, and international conferences, and commodity group meetings (). During the report period, the PI was author or coauthor on 31 invited or keynote presentations, as well as numerous submitted oral presentations and posters. Presentations were made at regional meetings (California Almond Board and California Pistachio Commission meetings, annual meetings of the Pacific Branch of the Entomological Society of America, invited lectures at universities), national meetings (annual meeting of the Entomological Society of America), and international meetings (annual meeting of the International Society for Chemical Ecology, European Congress of Entomology, 2nd World Life Sciences Congress, IOBC meetings). What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Impact: Activities, outcomes, and products have resulted in the following advances for protection of North American crops, orchards, forests, and natural and urban landscapes from invasive pests: 1. We identified, synthesized, and developed protocols for use of pheromone-based attractants for several hundred species of wood-boring beetles (Cerambycidae). These insects are a major target for surveillance because they are frequently moved between continents by global commerce. Much of the technology has been transferred to industry, and the attractants are now widely used by state and federal regulatory agencies (e.g., USDA-APHIS and CAPS), as well as analogous agencies in other countries. 2. We identified an attractant for the invasive Giant African Land snail, which has been the target of continuous eradication efforts in Florida for ~10 years. Our attractant is being used in the Florida eradication program. We also developed a powerful generic attractant for pest snails and slugs; this work is being submitted for publication. 3. We assisted in successful eradication of invasive red palm weevil from California. 4. We identified the first pheromonal attractants for North American wireworms, pests of increasing importance for cereals, root vegetables, and sugar cane. Some of these pheromones are being developed for areawide monitoring of pest species in the southeastern US. The project is being expanded to other wireworm pests of US agriculture. 5. We identified the key compound that the invasive bagrada bug uses to locate and attack seedlings of plants in the Brassica family. 6. With collaborators, we identified compounds that the queens of insects that live in large colonies, such as ants and wasps, use to maintain control of their nests. These compounds prevent female workers from becoming reproductively active, and have potential to be used for the disruption of the colony organization as an ecofriendly method of pest control. Overall, this project has produced >50 refereed scientific articles, and results have been broadly disseminated at regional, national, and international conferences. Accomplishments by objective: Objective 1. To identify, synthesize, and develop semiochemicals used by native and exotic cerambycid beetles. During the project period, pheromones or likely pheromones were identified for several hundred cerambycid species, including species in the genera Aromia, Trichoferus, Semanotus, Callidium, Arhopalus, and Xylotrechus which have already invaded North America or other continents, and well as numerous other cerambycids with the potential to be invasive pests based on their host ranges and biology. In part, this large number of identifications was possible because we showed that pheromone structures are often conserved among related cerambycid species, even those on different continents. Thus, our strategy of field screening every new pheromone that we identify in different countries produced many leads for pheromones for new species. This work was only possible because of our efforts to build a network of collaborators in foreign countries who are willing to field test pheromones for us. This has greatly leveraged project funds and resources. Because of the large number of pheromones now identified, we have also increased our predictive power, in terms of being able to predict what type of pheromone a new invader is likely to use based on its taxonomic position, and the likely chemistry of that pheromone. This in turn will accelerate identification of pheromone-based monitoring tools for any cerambycids that do invade, which in turn will greatly increase chances of eradication. In short, we have provided regulatory agencies with numerous new tools for surveillance, detection, and eradication of invasive cerambycid beetles. Objective 2. To proactively identify semiochemicals of other exotic pests that are likely to be introduced into the US through global commerce, and develop lures that can be used for their detection. In 2015, we initiated a study on the sex attractant pheromones of North American wireworm species. Although these species are endemic rather than invasive, they are rapidly becoming significant problems in many crop systems due to changes in insecticide use. No pheromones were known from any North American species; two reports of pheromone identifications from the 1970s have proven to be incorrect. In 2016, we obtained the first major positive results from this study, with pheromones being identified from 5 species in different genera. The importance of these results cannot be overemphasized: these represent the first correct identifications of pheromones for any of the >1,000 species of North American wireworm species, some of which are major agricultural pests. During the project period, working with USDA and university-based collaborators in Washington, Idaho, Illinois, South Carolina, and British Columbia, we identified likely pheromones from species in the genera Melanotus, Elator, Dalopius, and Parallelostethus, and published the pheromone of Melanotus communis, an important pest of potato and other root crops in the eastern US. Field trials with these compounds and a series of analogs are ongoing. Overall, this research is rapidly picking up momentum, and our collaborators will be shipping additional target species to us for pheromone identifications this year. During the project period, we published a first paper on the isolation of a powerful attractant for the invasive bagrada bug, Bagrada hilaris, from cauliflower seedlings. We have now identified this novel compound, given the common name brassicadiene, as a tricyclic sesquiterpene hydrocarbon, and the manuscript describing its identification has been submitted for publication. Objective 3: To explore the role of contact pheromones as mediators of insect behavior, and where feasible, develop methods for exploiting those signals for insect management. We are collaborating with several groups on various aspects of contact pheromones. First, work with Prof. Ken Ross in Georgia is identifying the chemical signals that distinguish monogyne (single queen) from polygyne (multiple queens) colonies of fire ants, a major pest across the southern US. Second, we are working with Olav Rueppell at NC State to identify CHC markers that identify honeybees as being diseased. Third, we have initiated a collaboration with Etya Ansalem at Penn State to study contact semiochemicals in bumblebee pollinators. We also collaborated with several international groups in efforts to identify queen pheromones for social insects, including ants, paper wasps, and yellowjackets. The overall goal of these and other ongoing collaborations is to gain a better understanding of how these signaling systems control the organization of colonies which can total millions of individuals. Our studies may identify signals or behaviors which can be exploited to disrupt colony organization, resulting in partial or complete control of species which are pests. Objective 4.To conduct preliminary studies on newly discovered invasive species, both to get a head start on developing management programs, and to obtain preliminary data to drive grant proposals. Working with Prof. Rory McDonnell at Oregon State University and Amy Roda, USDA-ARS in Florida, we developed an essential oil-based attractant for the highly invasive Giant African land snail, Lissachatina fulica. This attractant has been incorporated into the ongoing eradication program for the snail in Florida, to improve detection of residual infestations after molluscidal treatments have been made. We have also discovered a more generic and more powerful attractant for this invasive species, and a number of other invasive gastropod species, including Cornu aspersum, and the invasive slug, Deroceras reticulatum. This work is currently being written up for publication.

Publications

• Type: Journal Articles Status: Published Year Published: 2018 Citation: 331. Kevin J. Loope, Jocelyn G. Millar, Erin Wilson Rankin. 2018. Weak nestmate discrimination behavior in native and invasive populations of a yellowjacket wasp (Vespula pensylvanica). Biological Invasions pp 1-14. https://doi.org/10.1007/s10530-018-1783-3
• Type: Journal Articles Status: Published Year Published: 2018 Citation: 333. Weliton D. Silva, Jos� Maur�cio S. Bento, Lawrence M. Hanks, Jocelyn G. Millar. 2018. (Z)-7-Hexadecene is an aggregation-sex pheromone produced by males of the South American cerambycid beetle Susuacanga octoguttata. J. Chemical Ecology 44: 1115-1119
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Salvatore Guarino, Mokhtar Abdulsattar Arif, Jocelyn G. Millar, Stefano Colazza, Ezio Peri. 2018. Novel diterpenes emitted by seedlings of Brassica species provide host location cues to Bagrada hilaris. PLoS One. 13(12): e0209870. https://doi.org/10.1371/journal.pone.0209870
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 341. Zolt�n Imrei, Mikael A. Molander, Inis B. Winde, Zs�fia Lohonyai, �va B�lintn� Csonka, J�zsef Fail, Lawrence M. Hanks, Yunfan Zou, Jocelyn G. Millar, Mikl�s T�th, Mattias C. Larsson. 2019. Identification of the aggregation-sex pheromone of Plagionotus arcuatus ssp. arcuatus (Coleoptera: Cerambycidae) from two geographically separated populations. Naturwissenschaften 106, 18:9. https://doi.org/10.1007/s00114-019-1612-2
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 342. Yunfan Zou, Laurel Hansen, Tian Xu, Steven A. Teale, Dong Hao, and Jocelyn G. Millar. Optimizing pheromone-based lures for the invasive red-necked longhorn beetle, Aromia bungii Journal of Pest Science 92: 1217-1225. https://doi.org/10.1007/s10340-019-01108-6
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 343. Robert Maxwell Collignon; Sean Halloran; Jacqueline M. Serrano; J. Steven McElfresh; Jocelyn G. Millar. 2019. An Unstable Monoterpene Alcohol as a Pheromone Component of the Longhorned Beetle Paranoplium gracile (Coleoptera: Cerambycidae)" Journal of Chemical Ecology 45:339-347. https://doi.org/10.1007/s10886-019-01063-7
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 344. Cintia Akemi Oi, Ricardo Caliari Oliveira, Jelle S. van Zweden, Sidnei Mateus, Jocelyn G. Millar, Fabio S. Nascimento, Tom Wenseleers. 2019. Do primitively eusocial wasps use queen pheromones to regulate reproduction? A case study of the paper wasp Polistes satan. Frontiers in Ecology and Evolution, Front. Ecol. Evol., 11 June 2019| https://doi.org/10.3389/fevo.2019.00199
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 345. Linnea R. Meier " Jocelyn G. Millar " Judith A. Mongold-Diers " Lawrence M. Hanks. 2019. (S)-Sulcatol is a Pheromone Component for Two Species of Cerambycid Beetles in the Subfamily Lamiinae. Journal of Chemical Ecology 45: 447-454
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 347. Mikael A. Molander1* " Bj�rn Eriksson1 " Inis B. Winde1 " Yunfan Zou2 " Jocelyn G. Millar2 " Mattias C. Larsson. 2019. The Aggregation-sex Pheromones of the Cerambycid Beetles Anaglyptus mysticus and Xylotrechus antilope ssp. antilope: New Model Species for Insect Conservation through Pheromone-based Monitoring. Chemoecology 29: 111-124
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 348. Robert M. Collignon; Jonathan A. Cale; Jocelyn G. Millar. 2019. Dose and density effects on the use of aggregation-sex pheromones by longhorn beetles (Coleoptera: Cerambycidae), and their possible role in optimal density strategy. Journal of Chemical Ecology 45: 217-226
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 349. Jacqueline M. Serrano, J. Steven McElfresh, Yunfan Zou, and Jocelyn G. Millar. 2019. Identification of Aggregation-Sex Pheromone Components for a Living Fossil, the False Click Beetle, Palaeoxenus dohrni Horn (Coleoptera: Eucnemidae). Journal of Chemical Ecology 45: 366-370. doi.org/10.1007/s10886-019-01068-2
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 350. Ann M. Ray, Joseph A. Francese, Yunfan Zou, Kristopher Watson, Damon J. Crook, Jocelyn G. Millar. 2019. Isolation and identification of a male-produced aggregation-sex pheromone for the velvet longhorned beetle, Trichoferus campestris (Faldermann). Scientific Reports, 9:4459 | https://doi.org/10.1038/s41598-019-41047-x
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 352. Mikael A. Molander, Inis B. Winde, Joseph Burman, Franklin N. Nyabuga, Tobias U. T. Lindblom, Lawrence M. Hanks, Jocelyn G. Millar, Mattias C. Larsson. 2019. Common cerambycid pheromone components as attractants for longhorn beetles (Cerambycidae) breeding in ephemeral oak substrates in northern Europe. Journal of Chemical Ecology 45: 537-548
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 353. Hanks LM, Mongold-Diers JA, Mitchell RF, Zou Y, Wong JCH, Meier LR, Johnson TD, Millar JG. 2019. The role of minor pheromone components in segregating 14 species of cerambycid beetles of the subfamily Cerambycinae. Journal of Economic Entomology 112: 2236-2252
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 357. Alenka }uni?-Kosi,Nataaa Stritih-Peljhan, Yunfan Zou, J. Steven McElfresh, and Jocelyn G. Millar A male-produced aggregation-sex pheromone of the beetle Arhopalus rusticus (Coleoptera: Cerambycidae, Spondylinae) may be useful in managing this invasive species. Scientific Reports 9:19570 | https://doi.org/10.1038/s41598-019-56094-7
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 354. Amy Roda; Jocelyn G. Millar; Christopher Jacobsen; Robin Veasey; Lenny Fujimoto; Arnold Hara; Rory McDonnell. 2019. A new synthetic lure for management of the invasive giant African snail, Lissachatina fulica. PLoS One 14(10):e0224270.https://doi.org/10.1371/journal.pone.0224270

Progress 10/01/17 to 09/30/18

Outputs
Target Audience:Audiences targeted and reached by this project included: 1. Regulatory officials working in local, state, national, and international agencies charged with crop protection and with detection and management of invasive species. 2. Scientists and regulatory officials in foreign countries, with mutual interests in minimizing damage caused by invasive species through development of better methods of detection, monitoring, and eradication of invasive insect and gastropod species. 3. Academics and research scientists working in chemical ecology and entomology, with an emphasis of applying chemical ecology to protection of crops and natural habitats. 4. Agricultural industry/commodity boards and related grower groups via talks at commodity group meetings, and one on one discussions with commodity board officials and grower representatives. 5. The general public through open house days, insect fairs, and related outreach type events. Changes/Problems:As described in previous sections, we have expanded our intended studies of pheromones for invasive species (Objective 2) to also include economically important native species, specifically wireworms that damage a wide variety of crops, including grains, potatoes, sugar beets, and sweet potatoes, among others. What opportunities for training and professional development has the project provided?Opportunities for training and professional development included: 1. Ongoing training and mentoring of two women PhD students in Entomology, one of whom is a Hispanic woman who is the first in her family to go to college, and one gay male PhD student in Chemistry. 2. Sent students to regional, national, and international conferences, where they gave presentations. Conferences included the annual meetings of the Pacific branch of the Entomological Society of America, the joint meeting of the Entomological Societies of America and Canada in Vancouver, and the annual meeting of the International Society of Chemical Ecology in Budapest. 3. Hosted a visiting student from Brazil, and a visiting scientist from China. How have the results been disseminated to communities of interest?Results have been disseminated to communities of interest through: 1. Nineteen publications including a book chapter, 16 articles in referred academic journals, and two conference proceedings. 2. Presentations at regional, national, and international conferences and commodity group meetings (California Almond Board and California Pistachio Commission) by myself and members of my group. During the report period, the PI was author or coauthor on 12 invited and 18 submitted presentations. What do you plan to do during the next reporting period to accomplish the goals?Work planned for the next reporting period, by objective: Objective 1. To identify, synthesize, and develop semiochemicals used by native and exotic cerambycid beetles. Over the winter, we have been replenishing our supplies of a number of cerambycid pheromones which are not commercially available, in anticipation of an active field season of testing newly identified compounds in different areas of the world, with the help of our network of collaborators. Specifically, because cerambycid pheromones appear to be highly conserved among closely related taxonomic groups, pheromones identified for a species in genus XX in North America may also be pheromone components for species in the same genus on other continents, and vice versa, even though the species have been separated for millions of years. We have already used this screening approach to tentatively identify or obtain leads on pheromones of numerous new species, which has greatly expanded our knowledge and understanding of pheromone use within the whole, large cerambycid family. We are also working to identify a number of entirely novel cerambycid pheromone structures, including compounds from South American, European, and Asian species, supplied to us in the form of crude extracts by collaborators on those continents. As an indication of how productive this work has been and continues to be, since 2004, we and our network of collaborators have identified pheromones or likely pheromones for several hundred species from all continents except Antarctica, including for a number of invasive species which are already in the US, and other species which are deemed at high risk of invading through international commerce in wood and wood products. Development of these pheromones is providing effective new tools for US regulatory agencies to detect exotic species if they do invade, and help in the eradication of localized populations before they have a chance to spread. A number of pheromone products companies are now carrying cerambycid pheromone product lines, in large part as a result of our research. Objective 2.To proactively identify semiochemicals of other exotic pests that are likely to be introduced into the US through global commerce, and develop lures that can be used for their detection. In 2016, we established the groundwork and obtained funding for a multiyear study to identify attractants for invasive slug and snail species. This project has now successfully identified two different crude attractants for invasive slug and snail species. During the coming year, we will be focusing on reconstructing the odor profile of the most effective and most generic of these two crude attractants, with the goal of developing lures with a clearly defined formulation, i.e., that represent a standardized and reproducible bait, as opposed to crude attractant sources where the attraction can be quite variable due to different ages and qualities of the crude material. Fueled by our recent successes in identifying and bioassaying the first pheromones for North American click beetle species, including the major pest Melanotus communis, we will expand our studies to include species from different genera and tribes. Although these insects are native, they are of increasing importance as crop pests, particularly in grains, corn, and root vegetables, in part because the new generation of insecticides is proving to be relatively ineffective against these insects. This project will be carried forward with collaborators in Washington, Montana, Illinois, South Carolina, and Canada. Objective 3: To explore the role of contact pheromones as mediators of insect behavior, and where feasible, develop methods for exploiting those signals for insect management. We have ongoing collaborations with groups at UC Riverside, University of Arizona, North Carolina State University, and University of Georgia, as well as several groups overseas, studying the role of queen pheromones and other cuticular lipids in fire ants, yellowjacket wasps, and honeybees. Our role in these studies is to isolate, identify, and synthesize the bioactive compounds, so that the full details of the required components of the active pheromones, and the functional roles of specific individual compounds (as opposed to crude extracts or fractions of extracts of variable composition), can be worked out. This in turn may provide leads for developing methods of controlling pest social insect species, and particularly invasive species, by manipulation or disruption of the queen signals, or signals indicating nestmate status or caste. Objective 4.To conduct preliminary studies on newly discovered invasive species, both to get a head start on developing management programs, and to obtain preliminary data to drive grant proposals. We have initiated a collaboration with researchers in New Zealand to study pheromones of invasive wasp species in the genus Vespula, focusing on North American pest species that have invaded New Zealand and Hawaii. Because these species (e.g., Vespula germanica and Vespula pensylvanica) are pests in both native and invaded ranges, anything that we discover should be widely applicable for their management and control worldwide. To date, in analyses of extracts of V. pensylvanica, we have found a novel compound that is specifically associated with virgin queens, i.e., a possible candidate for a queen sex pheromone. This work is on hold until wasp colonies become active again in spring.

Impacts
What was accomplished under these goals? Accomplishments by objective: Objective 1. To identify, synthesize, and develop semiochemicals used by native and exotic cerambycid beetles. During the report period, work was focused on synthesis and field testing of recently identified pheromones to develop practical protocols for their use, and the identification and synthesis of entirely new pheromones. In North America, work included studies on species in the genera Callidiellum, Semanotus, Paranoplium, and Graphisurus among others, and the invasive species Trichoferus campestris, in collaboration with USDA-APHIS personnel. With the help of our network of international collaborators, field trials were conducted in South America with species in the genera Sussuacanga, Achryson, and Lophopoeum, in Europe with species in the genera Xylotrechus, Aromia, Rosalia, Molorchus, Obrium, and Trichoferus, among others, and in Asia with species in the genera Chlorophorus and Aromia, among others. Some of this work has been published, and most of the rest will be submitted for publication during the current cycle. Overall, our goal is to provide regulatory agencies, forest managers, and other end users with powerful new tools for detection and management of both native and exotic cerambycid pest species. Objective 2. To proactively identify semiochemicals of other exotic pests that are likely to be introduced into the US through global commerce, and develop lures that can be used for their detection. Over the past year, we have been collaborating with Prof. Stefano Colazza (University of Palermo, Italy) on the identification of a novel diterpene hydrocarbon produced by seedlings of Brassica plants which is a powerful attractant for the highly invasive bagrada bug, Bagrada hilaris. This bug invaded California about 10 years ago, and rapidly spread through the southwestern states, where it caused major damage to broccoli, cauliflower, and other brassicaceous crops. My group isolated and identified the attractant from crude extracts sent by Colazza, and a first paper has been published. In 2015, we initiated a study on the sex attractant pheromones of North American wireworm species. Although these species are endemic rather than invasive, they are rapidly becoming significant problems in many crop systems due to changes in insecticide use. No pheromones were known from any North American species; two reports of pheromone identifications from several decades ago have proven to be incorrect. To date, we have identified the pheromones for 5 native click beetle species in three genera, including the agriculturally important corn wireworm Melanotus communis. As a direct result of these pilot studies supported by Hatch funds, I was recently invited to join a multistate grant proposal aimed at identifying and field testing pheromones for click beetles infesting potatoes and sweet potatoes, that is anticipated to include scientists from Washington, Idaho, California, Illinois, and South Carolina. To date, we have published one paper on this work, with two more submitted and another in preparation. The importance of these results cannot be overemphasized: these represent the first correct identifications of pheromones for any of the >1,000 species of North American wireworm species, some of which are major agricultural pests. Now that we have gotten our foot in the door, and because pheromone structures are usually conserved within genera and sometimes even subfamilies, we anticipate that continuing studies with these compounds and close analogs will rapidly result in a number of additional pheromone identifications, providing the first simple and really practical tools for monitoring North American pest wireworm species. In anticipation of continuing to collaborate with scientists across the US on these important pests, I have obtained the appropriate permits allowing shipment of native wireworm adults into the UCR quarantine facility from anywhere in the continental US. Objective 3: To explore the role of contact pheromones as mediators of insect behavior, and where feasible, develop methods for exploiting those signals for insect management. My group is collaborating with several different groups on various aspects of contact pheromones, with a particular focus on their roles in social insects. During the report cycle, we finished up a project with Prof. Erin Wilson-Rankin at UCR, aimed at determining whether the cuticular hydrocarbons used for nestmate recognition are altered in invasive populations in Hawaii, as a result of going through a genetic bottleneck during invasion. This work has been published. Second, we are working with Prof. Ken Ross in Georgia, with the goal of determining the chemical signals that distinguish monogyne (single queen) from polygyne (multiple queens) colonies of fire ants, a major pest across the southern US, and which invaded California about 15 years ago. Third, we are collaborating with Olav Rueppel at NC State on a project to identify CHC markers that identify honeybees as being diseased. Fourth, we are collaborating with Tom Wenseleers (Catholic University of Leuven, Belgium) on the identification of new chemical classes of pheromones that honeybee queens use to maintain the division of labor within honeybee colonies. The overall goal of these and other ongoing collaborations with entomologists studying social insects is to gain a better understanding of how these chemical signaling systems work in controlling the organization of colonies which can total millions of individuals. Our joint studies may identify signals or behaviors which can be exploited to disrupt colony organization, resulting in partial or complete control of species which are pests, without having to resort to insecticides. Objective 4.To conduct preliminary studies on newly discovered invasive species, both to get a head start on developing management programs, and to obtain preliminary data to drive grant proposals. Working with Prof. Rory McDonnell at Oregon State University and Amy Roda, USDA-APHIS in Miami, we have identified two novel sources of attractants for invasive snail species. One of these appears to be a generic attractant for many species of invasive slugs and snails, as evidenced by bioassays with populations of invasive gastropod species in Hawaii, Oregon, Montana, Florida, and Trinidad. We are currently working to analyze and reconstruct the blend of volatile components that constitutes the active attractant blend, i.e., the minimum blend of odor compounds which is both necessary and sufficient for good attraction. The laboratory and field bioassay data collected so far with the crude attractants will provide the basis for at least two publications, and the preliminary data required to drive a full federal grant proposal.

Publications

• Type: Book Chapters Status: Published Year Published: 2018 Citation: Donald C. Weber, Ashot Khrimian, Maria C. Blassioli-Moreas, and Jocelyn G. Millar. 2018. Semiochemistry of Pentatomoidea, Chapter 14 in Biology of the Pentomoidea, Stink Bugs and Their Relatives. Eds. Jay McPherson and Thomas Miller. Taylor and Francis. Pp. 677-725
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Jesse D. Slone, Gregory M. Pask, Stephen T. Ferguson, Jocelyn G. Millar, Shelley L. Berger, Danny Reinberg, J�rgen Liebig, Anandasankar Ray and Laurence J. Zwiebel. 2017. Functional Characterization of Odorant Receptors in the Ponerine Ant, Harpegnathos saltator. Proceedings of the National Academy of Sciences of the USA 114: 8586-8591
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Alenka }uni? Kosi, Yunfan Zou, Michael Hoskovec, Al Vrezec, Nataaa Stritih, Jan Borucky, Jocelyn G. Millar. Novel, male-produced aggregation pheromone of the cerambycid beetle Rosalia alpina, a priority species of European conservation concern. PLoS ONE 12(8):e0183279. https://doi.org/10.1371/journal.pone.0183279
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Jocelyn G. Millar, Robert F. Mitchell, Judith A. Mongold-Diers, Yunfan Zou, Carlos E. Bogr�n, Melissa K. Fierke, Matthew D. Ginzel, Crawford W. Johnson, James R. Meeker, Therese M. Poland, Iral Ragenovich, and Lawrence M. Hanks. 2018. Identifying possible pheromones of cerambycid beetles by field testing known pheromone components in four widely separated regions of the United States. Journal of Economic Entomology 111:252-259. doi: 10.1093/jee/tox312
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Jocelyn G. Millar, Robert F. Mitchell, Linnea R. Meier, Todd D. Johnson, Judith A. Mongold-Diers, and Lawrence M. Hanks. 2017. (2E,6Z,9Z)-2,6,9-Pentadecatrienal as a male-produced aggregation-sex pheromone of the cerambycid beetle Elaphidion mucronatum. Journal of Chemical Ecology 43: 1056-1065
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Maria Cordoba, Jocelyn G. Millar, Rory McDonnell. 2018. Development of a high throughput laboratory bioassay for testing potential attractants for terrestrial snails and slugs. Journal of Economic Entomology 111: 637-644
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Jocelyn G. Millar, Austin B. Richards, Sean Halloran, Yunfan Zou, Elizabeth A. Boyd, Kristin N. Quigley, and Lawrence M. Hanks. 2018. Pheromone identification by proxy: identification of aggregation-sex pheromones of North American cerambycid beetles in the genera Callidium and Semanotus as a strategy to identify pheromones of invasive Asian congeners. Invited paper for a Special Issue of the Journal of Pest Science. https://doi.org/10.1007/s10340-018-0962-4
• Type: Journal Articles Status: Published Year Published: 2018 Citation: L. M. Hanks, J. A. Mongold-Diers, T. H. Atkinson, M. K. Fierke, M. D. Ginzel, E. E. Graham, T. M. Poland, A. B. Richards, M. L. Richardson, and J. G. Millar. 2018. Blends of pheromones, with and without host plant volatiles, can attract multiple species of cerambycid beetles simultaneously. Journal of Economic Entomology 111: 716-724
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Jacqueline M. Serrano, R. Maxwell Collignon, Yunfan Zou, and Jocelyn G. Millar. 2018. Identification of sex pheromones and sex pheromone mimics for two North American click beetle species (Coleoptera: Elateridae) in the genus Cardiophorus Esch. Journal of Chemical Ecology, 44: 327-338
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Les Greenberg, Christine Johnson, James Trager, J. Steven McElfresh, Joshua Rodstein, and Jocelyn G. Millar. 2018. Sex attractant pheromones of virgin queens of two sympatric slave-making ant species in the genus Polyergus and their possible roles in reproductive isolation. Journal of Chemical Ecology 44:547-555
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Jianting Fan, Olivier Denux, Claudine Courtin, Alexis Bernard, Marion Javal, Jocelyn G. Millar, Lawrence M. Hanks, Alain Roques. 2018. Multi?component blends for trapping native and exotic longhorn beetles at potential points?of?entry and in forests. Invited article, special issue of Journal of Pest Science. https://doi.org/10.1007/s10340-018-0997-6
• Type: Journal Articles Status: Published Year Published: 2018 Citation: S. Halloran, R. M. Collignon, J. S. McElfresh, and J. G. Millar. 2018. Fuscumol and geranylacetone as pheromone components of Californian longhorn beetles (Coleoptera: Cerambycidae) in the subfamily Spondylidinae. Environmental Entomology 47: 1300-1305
• Type: Journal Articles Status: Published Year Published: 2018 Citation: L. Saul-Gershenz, J. G. Millar, J. S. McElfresh, N. M. Williams. 2018. Deceptive signals and behaviors of a cleptoparasitic beetle show local adaptation to different host bee species. Proceedings of the National Academy of Sciences of the USA 115: 9756-9760
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Robert F. Mitchell, Ann M. Ray, Lawrence M. Hanks, and Jocelyn G. Millar. 2018. The common natural products (S)-?-terpineol and (E)-2-hexenol are important pheromone components of the cerambycid beetle Megacyllene antennata. Environmental Entomology 47: 1547-1552
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Mikael A. Molander, Jimmy Helgesson, Inis B. Winde, Jocelyn G. Millar, Mattias C. Larsson. 2019. The male-produced aggregation-sex pheromone of the endangered cerambycid beetle Plagionotus detritus ssp. detritus. Journal of Chemical Ecology 45: 28-36
• Type: Journal Articles Status: Accepted Year Published: 2019 Citation: Bridget Bobadoye, Baldwyn Torto, Ayuka Fombong, Yunfan Zou, Karl Adlbauer, Lawrence M. Hanks, and Jocelyn G. Millar. 2019. Evidence of Aggregation-sex Pheromone Use by Cerambycid Beetles Species Native to Africa. Environmental Entomology
• Type: Journal Articles Status: Accepted Year Published: 2019 Citation: Ann M. Ray, Joseph A. Francese, Yunfan Zou, Kristopher Watson, Damon J. Crook, Jocelyn G. Millar. 2019. Isolation and identification of a male-produced aggregation-sex pheromone for the velvet longhorned beetle, Trichoferus campestris (Faldermann). Scientific Reports
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Lawrence M. Hanks1 and Jocelyn G. Millar. 2018. Conservation of pheromone chemistry within the Cerambycidae on a global scale, and implications for invasion biology. Proc. 29th USDA Interagency Research Forum on Invasive Species 2016. Jan. 10-12, 2018, Annapolis MD
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Jocelyn G. Millar and Lawrence M. Hanks. 2018. Practical applications for pheromones of cerambycid beetles. Seminar proceedings, Interactions behind the pine wilt disease. Instituto Nacional de Invesigacao Agraria e Veterinaria. June 21, 2018. Oeiras, Portugal

Progress 10/01/16 to 09/30/17

Outputs
Target Audience:Audiences targeted and reached by this project included: 1. Academics and research scientists working in chemical ecology and entomology, with an emphasis of applying chemical ecology to protection of crops and natural habitats. 2. Regulatory officials working in local, state, national, and international agencies charged with crop protection and with detection and management of invasive species. 3. Agricultural industry/commodity boards and related grower groups via talks at commodity group meetings, and one on one discussions with commodity board officials and grower representatives. 4. The general public through open house days at the University. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?1. Ongoing training and mentoring of two PhD students in Entomology, one of whom is a Hispanic woman who is the first in her family to go to college, and one PhD student in Chemistry. 2. Working with a Hispanic woman postdoctoral scholar. 3. Summer internship for an undergraduate student 4. Sending students to regional and national conferences, where they gave presentations. Conferences included the annual meetings of the Pacific branch of the Entomological Society of America and the Entomological Society of America national meeting. 5. Mentoring a PhD student in submitting bids to organize symposia for two international meetings. 6. Lectured and taught at a chemical ecology short-course for grad students and postdocs, including two days of conducting hands-on training in the laboratory with the attendees. How have the results been disseminated to communities of interest?Results have been disseminated to communities of interest through: 1. Ten publications including a review article and book chapter, and 8 articles in referred academic journals. 2. Presentations at regional, national, and international conferences, and commodity group meetings (California Almond Board and California Pistachio Commission). During the report period, the PI was author or coauthor on 7 invited and 13 submitted presentations. 3. Websites and online reporting services which have developed stories based on our work with e.g., identification of cuticular hydrocarbon receptors in ants. What do you plan to do during the next reporting period to accomplish the goals?Work planned for the next reporting period, by objective: Objective 1. To identify, synthesize, and develop semiochemicals used by native and exotic cerambycid beetles. We are currently scaling up and streamlining the syntheses of several newly identified cerambycid pheromones so that we can make them available to collaborators around the world for testing. Specifically, because cerambycid pheromones appear to be highly conserved among closely related taxonomic groups, pheromones identified for a species in genus XX in North America may also be pheromone components for species in the same genus on other continents, even though the species have been separated for millions of years. We have already used this screening approach to tentatively identify or at least obtain leads on numerous new species, which has greatly expanded our knowledge and understanding of pheromone use within the whole, large cerambycid family, which comprises >35,000 described species. We are also working to identify a number of entirely novel cerambycid pheromone structures, including 4 from South American species and two from North American species. During the coming field season, in addition to our ongoing collaborations with research groups in Europe, Japan, China, northern Brazil, and Africa, we will be starting two new collaborations aimed at identifying pheromones of potentially invasive species, with research groups in southern Brazil and Hong Kong. As an indication of how productive this work has been and continues to be, since 2004, we and our network of collaborators have identified pheromones or likely pheromones for several hundred species from all continents except Antarctica, including for a number of invasive species which are already in the US, and other species which are deemed at high risk of invading through international commerce in wood and wood products. Development of these pheromones is providing effective new tools for US regulatory agencies to rapidly detect exotic species if they do, and help in the eradication of localized populations before they have a chance to spread. Objective 2.To proactively identify semiochemicals of other exotic pests that are likely to be introduced into the US through global commerce, and develop lures that can be used for their detection. In 2016, we established the groundwork and obtained funding for a multiyear study to identify attractants for invasive slug and snail species, with key milestones being the development of a high throughput bioassay and the identification of a consistently attractive food plant (manuscript in press, another in prep.). During the coming year, with our Oregon and Montana collaborators, we will attempt to identify the subset of compounds that is both necessary and sufficient to obtain strong and reproducible attraction of three invasive slugs and snails, as a first step towards developing effective and commercializable baits for these pests of numerous crops. Fueled by our successes in 2017 in conclusively identifying and bioassaying pheromones for five North American click beetle species, we will expand our studies to include species from different genera and tribes. Although these insects are native, they are of increasing importance as crop pests, particularly in grains, corn, and root vegetables, in part because the new generation of insecticides is proving to be relatively ineffective against these insects. This project will be carried forward with collaborators in Washington, Montana, Illinois, and South Carolina. Objective 3: To explore the role of contact pheromones as mediators of insect behavior, and where feasible, develop methods for exploiting those signals for insect management. We have ongoing collaborations with groups at UC Riverside, University of Arizona, North Carolina State University, and University of Georgia, as well as several groups overseas, studying the role of queen pheromones and other cuticular lipids in fire ants, trapjaw ants, yellowjacket wasps, and honeybees. Our role in these studies is to isolate, identify, and synthesize the bioactive compounds, so that the full details of the required components of the active pheromones, and their functional roles, can be worked out. This in turn may provide leads for developing methods of controlling pest social insect species, and particularly invasive species, by manipulation or disruption of the queen signals, or signals indicating nestmate status or caste. Objective 4.To conduct preliminary studies on newly discovered invasive species, both to get a head start on developing management programs, and to obtain preliminary data to drive grant proposals. We are initiating a collaboration with researchers in New Zealand to study pheromones of invasive wasp species in the genus Vespula, focusing on North American pest species that have invaded New Zealand and Hawaii. Because these species (e.g., Vespula germanica and Vespula pensylvanica) are pests in both native and invaded ranges, anything that we discover should be widely applicable for their management and control worldwide.

Impacts
What was accomplished under these goals? Objective 1. To identify, synthesize, and develop semiochemicals used by native and exotic cerambycid beetles. During the report period, we identified, synthesized, and field tested a number of new cerambycid pheromones from species in the genera Ambonus, Chlorida, and Cotyclytus from South America, and Dryobius and Elaphidion from North America, and results on all species have been published. We also synthesized large amounts of the pheromone for the invasive species Trichoferus campestris, an invasive pest which has spread to at least several other states, for use in trials aimed at optimizing operational protocols for use of the pheromone in monitoring the spread of this pest, and for testing against potentially invasive congeners in Europe. We also synthesized the enantiomers of the pheromone of Aromia bungii, which were field tested in Japan and China versus the racemic material. This species is a notorious pest of stone fruits which has recently invaded Japan and parts of Europe. USDA-APHIS has categorized this species as very high risk for the US. Identification and optimization of this and the other pheromones described above will provide regulatory agencies with powerful tools for detections of new incursions, and for helping in eradication efforts if populations of exotic cerambycids do become established in the US. Objective 2. To proactively identify semiochemicals of other exotic pests that are likely to be introduced into the US through global commerce, and develop lures that can be used for their detection. In 2015, we initiated a study on the sex attractant pheromones of North American wireworm species. Although these species are endemic rather than invasive, they are rapidly becoming significant problems in many crop systems due to changes in insecticide use. No pheromones were known from any North American species; two reports of pheromone identifications from several decades ago have proven to be incorrect. In 2016, we obtained the first major positive results from this study, with pheromones or likely pheromones being identified from 5 species in different genera. In 2017, we synthesized and field tested these pheromones, verifying their activity. The importance of these results cannot be overemphasized: these represent the first correct identifications of pheromones for any of the >1,000 species of North American wireworm species, some of which are major agricultural pests. Now that we have gotten our foot in the door, and because pheromone structures are usually highly conserved within genera and sometimes even subfamilies, we anticipate that continuing studies with these compounds and close analogs will result in a number of additional pheromone identifications, providing the first simple and really practical tools for monitoring pest wireworm species in North America. To maximize our reach and impact, we are working with collaborators in different climatic regions, including Washington, Illinois, and South Carolina, who are field testing pheromone candidates, and sending us live specimens for pheromone analyses. We have the appropriate permits allowing shipment of native wireworm adults into the UCR quarantine facility. Objective 3: To explore the role of contact pheromones as mediators of insect behavior, and where feasible, develop methods for exploiting those signals for insect management. My group is collaborating with several different groups on various aspects of contact pheromones. First, at UCR, we are working with Prof. Erin Wilson-Rankin to determine whether the cuticular hydrocarbons used for nestmate recognition is altered in invasive populations in Hawaii, as a result of going through a genetic bottleneck during invasion. Second, we are working with Prof. Ken Ross in Georgia, with the goal of determining the chemical signals that distinguish monogyne (single queen) from polygyne (multiple queens) colonies of fire ants, a major pest across the southern US, and which invaded California about 15 years ago. Third, in another study published this year in Nature Communications, we collaborated with a multidisciplinary group to identify the antennal receptors which detect the cuticular hydrocarbon signals that ants (and many other social insect species) use for discriminating nestmates from nonnestmates, and that signal sex, caste, and reproductive status. Fourth, we are collaborating with Olav Rueppel at NC State on a project to identify CHC markers that identify honeybees as being diseased. The overall goal of these and other ongoing collaborations with entomologists studying social insects is to gain a better understanding of how these chemical signaling systems work in controlling the organization of colonies which can total millions of individuals. Our joint studies may identify signals or behaviors which can be exploited to disrupt colony organization, resulting in partial or complete control of species which are pests, without having to resort to insecticides. Objective 4.To conduct preliminary studies on newly discovered invasive species, both to get a head start on developing management programs, and to obtain preliminary data to drive grant proposals. Working with Prof. Rory McDonnell at Oregon State University, we developed a high throughput bioassay method for assaying attractants for invasive snail and slug species (manuscript in press). With the bioassay, we screened a large number of food sources and other potential attractants, and found that of all odor sources tested, chopped cucumber was consistently the most attractive odor source, for an invasive snail and an invasive slug species. Preliminary field bioassays in Montana in an area which had been invaded by the invasive eastern heath snail Xerolenta obvia, verified the activity. Our joint efforts are now focused on reconstructing and bioassaying the odor of chopped cucumber, to identify the subset of compounds that are both necessary and sufficient for attraction. In parallel, we are testing the same suites of compounds against another invasive snail, Cornu aspersum, and an invasive slug, Deroceras reticulatum.

Publications

• Type: Journal Articles Status: Published Year Published: 2016 Citation: Larry Schmeelk, Jocelyn G. Millar, Lawrence M. Hanks. 2016. Influence of trap height and bait type on abundance and species diversity of cerambycid beetles captured in forests of east-central Illinois. Journal of Economic Entomology 109:1750-1757
• Type: Journal Articles Status: Published Year Published: 2016 Citation: G. P. Hughes, L. R. Meier, Y. Zou, J. G. Millar, L. M. Hanks, and M. D. Ginzel. 2016. Stereochemistry of Fuscumol and Fuscumol Acetate Influences Attraction of Longhorned Beetles of the Subfamily Lamiinae. Environmental Entomology 45: 1271-1275
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Jocelyn G. Millar1,*, Kenneth F. Haynes2, Aaron T. Dossey3, J. Steven McElfresh1 and Jeremy D. Allison. 2016. Sex attractant pheromone of the Luna moth, Actias luna (Linnaeus). Journal of Chemical Ecology 42:869-876.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: John M. Hash, Jocelyn G. Millar, John M. Heraty, James F. Harwood, and Brian V. Brown. 2017. Millipede defensive compounds are a double-edged sword: Natural history of the millipede-parasitic fly genus Myriophora Brown (Diptera: Phoridae). Journal of Chemical Ecology 43: 198-206
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Tian Xu, Hiroe Yasui, Stephen A. Teale, Nao Fujiwara-Tsujii, Jacob D. Wickham, Midori Fukaya, Laura Hansen, Satoshi Kiriyama, Dejun Hao, Akio Nakano, Long-Wa Zhang, Takahito Watanabe, Masahiko Tokoro, and Jocelyn G. Millar. Identification of a male-produced sex-aggregation pheromone for a highly invasive cerambycid beetle, Aromia bungii. Scientific Reports, June 8-17
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Gregory M. Pask, Jocelyn G. Millar, Jesse D. Slone, Prithwiraj Das, Jardel A. Moreira, Xiaofan Zhou, Jan Bello, Shelley L. Berger, Roberto Bonasio, Claude Desplan, Danny Reinberg, J�rgen Liebig, Laurence J. Zwiebel, and Anandasankar Ray. 2017. Receptors for cuticular hydrocarbons: Candidate pheromones and cues for social insects. Nature Communications 8:297DOI: 10.1038/s41467-017-00099-1.
• Type: Book Chapters Status: Published Year Published: 2017 Citation: Jocelyn G. Millar and Lawrence M. Hanks. 2017. Chapter 5 - Chemical ecology of Cerambycidae. In Q. Wang (ed. ) Cerambycidae of the World: Biology and Pest Management. Boca Raton: CRC Press/Taylor & Francis. Pp 161-208.
• Type: Other Status: Published Year Published: 2016 Citation: Lawrence M. Hanks and Jocelyn G. Millar. 2016. Sex and Aggregation-Sex Pheromones of Cerambycid Beetles: Basic Science and Practical Applications. In: July Special Issue of the Journal of Chemical Ecology on Practical Applications of Semiochemicals Journal of Chemical Ecology 42:631-654
• Type: Journal Articles Status: Published Year Published: 2017 Citation: J.C.H. Wong, Y. Zou, J. G. Millar, and L. M. Hanks. 2017. Attraction of adult cerambycid beetles to their aggregation-sex pheromones is influenced by volatiles from host plants of their larvae. Environmental Entomology 46:649-653
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Natalie M. Diesel, Yunfan Zou, Todd D. Johnson, Donald A. Diesel, Jocelyn G. Millar, Judith A. Mongold-Diers, and Lawrence M. Hanks. 2017. Identification of the male-produced aggregation-sex pheromone of the rare North American cerambycid beetle Dryobius sexnotatus. Journal of Chemical Ecology 43: 739-744

Progress 10/01/15 to 09/30/16

Outputs
Target Audience:Target audiences reached by this project included: 1. Regulatory officials working in local, state, national, and international agencies charged with crop protection and with detection and management of invasive species. 2. Academics and research scientists working in chemical ecology and entomology, with an emphasis of applying chemical ecology to protection of crops and natural habitats 3. Agricultural industry/commodity boards and related grower groups via talks at commodity group meetings, and one on one discussions with commodity board officials and grower representatives. 4. The general public through open house days at the University Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Opportunities for training and professional development included: 1. Ongoing training and mentoring of two PhD students in Entomology, one of whom is a Hispanic woman who is the first in her family to go to college. 2. Mentoring a Phd student for a successful NIFA predoctoral grant proposal 3. Mentoring a Hispanic woman postdoctoral scholar. 4. Summer internship for an undergraduate student 5. Hosting a female visiting scholar from Sweden for three months. 6. Sending students to regional and national conferences, where they gave presentations. Conferences included the annual meetings of the Pacific branch of the Entomological Society of America and the Entomological Society of America national meeting, run jointly with the International Congress of Entomology. How have the results been disseminated to communities of interest?Results have been disseminated to communities of interest through: 1. Eleven publications including a review article and book chapter (in press), and 9 articles in referred academic journals. This included obtaining cover photos on two journals. 2. Presentations at regional, national, and international conferences, lectures at other universities, and commodity group meetings. During the report period, the PI was author or coauthor on 10 invited and 23 submitted presentations. 3. Websites and online reporting services which have developed stories based on our work with e.g., queen pheromones of ants. What do you plan to do during the next reporting period to accomplish the goals?Work planned for the next reporting period, by objective: Objective 1. To identify, synthesize, and develop semiochemicals used by native and exotic cerambycid beetles. We are currently scaling up and streamlining the syntheses of several newly identified cerambycid pheromones so that we can make them available to collaborators around the world for testing. Specifically, because cerambycid pheromones appear to be highly conserved among closely related taxonomic groups, pheromones identified for a species in genus XX in North America may also be pheromone components for species in the same genus on other continents, even though the species have been separated for millions of years. We have already used this screening approach to tentatively identify or at least obtain leads on numerous new species, which has greatly expanded our knowledge and understanding of pheromone use within the whole, large cerambycid family, which comprises >30,000 described species. We are also working to identify 5 entirely novel cerambycid pheromone structures, including 3 from South American species and two from North American species. During the coming field season, we will also be optimizing the pheromone blends and operational parameters for use of pheromones for two important invasive species, Trichoferus campestris, which has already become established in parts of the US, and Aromia bungii, which has not yet invaded the US, but which has successfully invaded Japan and several countries in Europe, where it is already causing substantial damage to orchard crops. Having a powerful attractant pheromone available for the latter species should allow US regulatory agencies to rapidly detect the latter species if it does invade, and help in the eradication of localized populations before it has a chance to spread. Objective 2.To proactively identify semiochemicals of other exotic pests that are likely to be introduced into the US through global commerce, and develop lures that can be used for their detection. Over the past year, we established the groundwork and obtained funding for a multiyear study to identify attractants for invasive slug and snail species, with key milestones being the development of a high throughput bioassay and the identification of a consistently attractive food plant. During the coming year, we will analyze the odor profile of the plant, reconstruct it from synthesized pure compounds, and test the reconstruction and subsets of the reconstruction in laboratory and field bioassays. Our goal is to identify the subset of compounds that is both necessary and sufficient to obtain strong and reproducible attraction of pest slugs and snails, for incorporation into IPM programs for these pernicious pests which to date, have not been well studied. We will also continue to expand a project on the identification of pheromones of wireworm species, fueled by our successes last year in identifying pheromones or likely pheromones for five North American species. Although these insects are native, they are of increasing importance as crop pests, particularly in grains, corn, and root vegetables, in part because the new generation of insecticides is proving to be relatively ineffective against these insects. This project will be carried forward with collaborators in Washington, Montana, Illinois, and South Carolina. Objective 3: To explore the role of contact pheromones as mediators of insect behavior, and where feasible, develop methods for exploiting those signals for insect management. We have ongoing collaborations with groups at UC Riverside, University of Arizona, North Carolina State University, and University of Georgia, as well as several groups overseas, studying the role of queen pheromones and other cuticular lipids in Argentine ants, fire ants, trapjaw ants, yellowjacket wasps, and honeybees. Our role in these studies is to isolate, identify, and synthesize the bioactive compounds, so that the full details of the required components of the active pheromones, and their functional roles, can be worked out. This in turn may provide leads for developing methods of controlling pest social insect species, and particularly invasive species, by manipulation or disruption of the queen signals, or signals indicating nestmate status or caste.

Impacts
What was accomplished under these goals? Accomplishments by objective: Objective 1. To identify, synthesize, and develop semiochemicals used by native and exotic cerambycid beetles. During the report period, we identified, synthesized, and field tested 4 new cerambycid pheromones, including pheromones for Trichoferus campestris, an invasive pest which become established in Utah and has since been detected in several other states. As part of those studies, pheromone baited traps were deployed in Pennsylvania, resulting in the first record of T campestris being present in that state. We also identified and successfully field tested the pheromone for Aromia bungii, a notorious pest of stone fruits which USDA-APHIS has deemed as very high risk for the US. This will provide regulatory agencies with a sensitive surveillance tool, both for detections of new incursions, and for helping in eradication efforts if populations do become established in the US. In additional collaborative studies, we identified two more new pheromones from South American cerambycid species, further increasing the arsenal of tools available to regulatory agencies for detection of new invasive species. Overall, these ongoing studies have unequivocally demonstrated that cerambycid pheromones are sensitive and selective tools for detection and monitoring both native and exotic pest cerambycid species. Objective 2.To proactively identify semiochemicals of other exotic pests that are likely to be introduced into the US through global commerce, and develop lures that can be used for their detection. In 2015, we initiated a study on the sex attractant pheromones of North American wireworm species. Although these species are endemic rather than invasive, they are rapidly becoming significant problems in many crop systems due to changes in insecticide use. No pheromones were known from any North American species; two reports of pheromone identifications from several decades ago have proven to be incorrect. In 2016, we obtained the first major positive results from this study, with pheromones or likely pheromones being identified from 5 species in different genera. Because the pheromone structures are usually highly conserved within genera and sometimes even subfamilies, we expect that continuing studies in 2017 will result in a number of additional pheromone identifications, providing the first simple and really practical tools for monitoring pest wireworm species in North America. To maximize our reach and impact, we are working with collaborators in different climatic regions, including Washington, Montana, Illinois, and South Carolina, who will both field test pheromone candidates, and send us live specimens for pheromone analyses. We have the appropriate permits allowing shipment of native wireworm adults into the UCR quarantine facility. Objective 3: To explore the role of contact pheromones as mediators of insect behavior, and where feasible, develop methods for exploiting those signals for insect management. This work is continuing in several different projects. First, with collaborators in Georgia, we are studying the chemical signals which appear to distinguish monogyne (single queen) from polygyne (multiple queens) colonies of the fire ant, which is a major pest across the southern US, and which invaded California about 15 years ago. In another study submitted for publication, we collaborated with a multidisciplinary group to identify the antennal receptors which are responsible for detection and recognition of the cuticular hydrocarbons signals that ants (and many other social insect species) use for discriminating nestmates from nonnestmates, and that signal sex, caste, and reproductive status. The overall goal of these and other ongoing collaborations with entomologists studying social insects is to gain a better understanding of how these chemical signaling systems work in controlling the organization of colonies which can total millions of individuals. This information in turn may elucidate signals or behaviors which can be exploited to disrupt colony organization, resulting in partial or complete control of species which are pests without having to resort to insecticides. Objective 4.To conduct preliminary studies on newly discovered invasive species, both to get a head start on developing management programs, and to obtain preliminary data to drive grant proposals. Working with collaborators now at Oregon State university, we developed a high throughput bioassay method for assaying attractants for invasive snail and slug species. Using this bioassay, we screened a large number of food sources and other potential attractants, identifying one species of plant which is consistently highly attractive. We are currently analyzing and reconstructing the odor profile of that plant, as a first step towards developing attractant lures for detection, monitoring, and control of pest slug and snail species, with a particular focus on invasive species.

Publications

• Type: Journal Articles Status: Published Year Published: 2016 Citation: Yunfan Zou, Claire E. Rutledge, Kiyoshi Nakamuta, Chris T. Maier, Lawrence M. Hanks, Austin B. Richards, Emerson S. Lacey, and Jocelyn G. Millar. 2016. Identification of a Pheromone Component and a Critical Synergist for the Invasive Beetle Callidiellum rufipenne (Coleoptera: Cerambycidae). Environmental Entomology 45:216-222
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Jacob. D. Wickham, Jocelyn G. Millar, Lawrence M. Hanks, Yunfan Zou, Joseph C. H. Wong, Rhett D. Harrison, and Yi Chen. 2016. (2R,3S)-2,3-Octanediol, a female-produced sex pheromone of Megopis costipennis (Coleoptera: Cerambycidae: Prioninae). Environmental Entomology 45:223-228.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Jacob D. Wickham, Wen Lu, Zhenqiang Peng, Dongfeng Guo, Jocelyn G. Millar, Lawrence M. Hanks, and Yi Chen. 2016. Prionic acid: an effective sex attractant for an important pest of sugarcane, the cerambycid beetle Dorysthenes granulosus. Rapid communication, Journal of Economic Entomology 109:484-486.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Collignon, M. R., I. Swift, Y. Zou, J. S. McElfresh, L. M. Hanks, J. G. Millar. 2016. The influence of host plant volatiles on the attraction of longhorned beetles (Coleoptera: Cerambycidae) to pheromones. Journal of Chemical Ecology 42:215-229
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Kim Hung, Themis Michaelides, Jocelyn Millar, Astri Wayadande, and Alec Gerry. 2015. House fly attraction to insect honeydew. PloS one, 10(5), p.e0124746
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Rory McDonnell, Ju Yoo, Kenna Patel, Lissette Rios, Robert Hollingsworth, Jocelyn Millar, and Timothy Paine. 2015. Can essential oils be used as novel drench treatments for the eggs and juveniles of the pest snail Cornu aspersum in potted plants? Journal of Pest Science 89: 549-555
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Jacob D. Wickham, Wen Lu, Long-Wa Zhang, Yi Chen, Yunfan Zou, Lawrence M. Hanks, and Jocelyn G. Millar. 2016. Likely aggregation-sex pheromones of the invasive beetle Callidiellum villosulum, and the related Asian species Allotraeus asiaticus, Semanotus bifasciatus, and Xylotrechus buqueti (Coleoptera: Cerambycidae). Journal of Economic Entomology 109:2243-2246
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Mark S. Hoddle, Mohammed Alzubaidy, Christina D. Hoddle, John Kabashima, J. Nicholas Nisson, Jocelyn Millar, Kevin Hoffmann, and Monica Dimson. 2016. The palm weevil Rhynchophorus vulneratus is eradicated from Laguna Beach, California, USA. California Agriculture Sept., 2016 1-7. http://dx.doi.org/10.3733/ca.2016a0012
• Type: Other Status: Published Year Published: 2016 Citation: Lawrence M. Hanks and Jocelyn G. Millar. 2016. Sex and Aggregation-Sex Pheromones of Cerambycid Beetles: Basic Science and Practical Applications. In: July Special Issue of the Journal of Chemical Ecology on Practical Applications of Semiochemicals Journal of Chemical Ecology 42:631-654
• Type: Book Chapters Status: Awaiting Publication Year Published: 2017 Citation: Donald C. Weber, Ashot Khrimian, Maria C. Blassioli-Moreas, and Jocelyn G. Millar. 2017. Semiochemistry of Pentatomoidea, Chapter 14 in Biology of the Pentomoidea, Stink Bugs and Their Relatives. Eds. Jay McPherson and Thomas Miller. Taylor and Francis
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Adrian A. Smith, Jocelyn G. Millar, Lawrence M. Hanks, and Andrew V. Suarez. 2016. Comparative analysis of fertility signals and sex-specific cuticular chemical profiles of Odontomachus trap-jaw ants. Journal of Experimental Biology 219: 419-430

Progress 10/01/14 to 09/30/15

Outputs
Target Audience:Target audiences that this project aims to serve included: Regulatory officials working in local, state, national, and international agencies charged with crop protection Academics and research scientists working in chemical ecology and entomology, with an emphasis of applying chemical ecology to protection of crops and natural habitats Agricultural industry/commodity boards and related grower groups. The general public through open house days at the University, nontechnical publications, and presentations. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Opportunities for training and professional development included: 1. Ongoing training and mentoring of two PhD students in Entomology 2. Mentoring a Phd student for a successful NIFA predoctoral grant proposal 3. Sending my students to regional, national, and international conferences, where they gave presentations. Conferences included the annual meetings of the Pacific branch of the Entomological Society of America, the Entomological Society of America national meeting, and the International Society of Chemical Ecology meeting. 4. Hosting and training a visiting scientist from Slovenia for 3 weeks. 5. Collaborations with foreign scientists in Sweden, Hungary, France, Kenya, Brazil, Japan, and China, that included helping to develop protocols for field trials, and organizing and supplying materials for those field trials. How have the results been disseminated to communities of interest?Results have been disseminated to communities of interest through: 1. Publications in academic journals (9 manuscripts involving this project during the report period) and technical books (2 book chapters or reviews submitted during the report period, and currently in press). 2. Presentations at regional, national, and international conferences, and lectures at other universities. During the report period, the PI was author or coauthor on 7 invited and 17 submitted presentations. 3. Websites, blogs, and similar online reporting services which have developed stories based on our work with e.g., valley elderberry longhorn borer, and queen pheromones of ants. What do you plan to do during the next reporting period to accomplish the goals?Work planned for the next reporting period, by objective: Objective 1. To identify, synthesize, and develop semiochemicals used by native and exotic cerambycid beetles. We are currently working to identify three new pheromone compounds from South American ceramycid species that have the potential to be invasive. Once identified, the compounds will be synthesized and their activity verified in field trials by Brazilian collaborators. The compounds will also be tested at several sites in the US, because of the known, highly conserved pheromone structures within this family, i.e., these compounds could easily be pheromones for closely related North American species as well. With Chinese collaborators, we are also planning the targeted pheromone identification of several Asian species with a high risk of being invasive, including Aromia bungii, Massicus radei, and two Semanotus species. In an ongoing collaboration, USDA-APHIS personnel will field test another pheromone that we have recently identified and are currently synthesizing, for the invasive species Trichoferus campestris. Objective 2.To proactively identify semiochemicals of other exotic pests that are likely to be introduced into the US through global commerce, and develop lures that can be used for their detection. We are not currently working on any invasive insect species other than cerambycid beetles as described above. However, we are ramping up a project on the identification of pheromones of wireworm species, and this coming field season we will be testing a full library of all known wireworm pheromones, which we are currently synthesizing. These pheromones will also be tested by collaborators in Montana, Illinois, and South Carolina. Objective 3: To explore the role of contact pheromones as mediators of insect behavior, and where feasible, develop methods for exploiting those signals for insect management. We have ongoing collaborations with groups at UC Riverside, University of Arizona, North Carolina State University, and University of Georgia, studying the role of queen pheromones and other cuticular lipids in Argentine ants, fire ants, trapjaw ants, yellowjacket wasps, and honeybees. Our role in these studies is to isolate, identify, and synthesize the bioactive compounds, so that the full details of the required components of the active pheromones, and their functional roles, can be worked out. This in turn may provide leads for developing methods of controling pest social insect species by manipulation of the queen signals.

Impacts
What was accomplished under these goals? Impact statement: This project has produced substantial impacts on several topics that relate directly to the detection, management and control of insect populations, including invasive and endemic pests, as well as endangered species. Specifically, we have: 1. Identified, synthesized, and developed protocols for practical use of a number of powerful attractants for detection and monitoring of pest insects by end user clientele such as growers and regulatory agencies charged with keeping invasive species out of North America. 2. With collaborators, identified several natural chemical signals that queen bees and ants use to maintain control of their colonies. Knowing what these chemicals are and how they work provides the foundation for development of possible control methods based on disruption of the signals, and hence the colony organization. 3. Began a multistate investigation of the sex pheromones of wireworms, which are rapidly increasing in importance as agricultural pests due to the phasing out of many broad-spectrum insecticides. These insects can be pests in numerous crops, including potatoes, cereals, corn, and vegetable crops. Accomplishments by objective: Objective 1. To identify, synthesize, and develop semiochemicals used by native and exotic cerambycid beetles. During the report period, we identified and synthesized 6 entirely new pheromone structures from endemic and exotic cerambycid species. Because pheromone structures are highly conserved within closely related species, we have no shown that these 6 compounds are used as pheromone components by at least 12 other species, and that number is certain to be a very low estimate of the total number of species that use these compounds as pheromones. Field trials with these and previously identified cerambycids demonstrated what powerful tools these compounds are for detection of cerambycids in surveillance programs, and delineation of their ranges. For example, a study of species richness and phenology conducted in Delaware with our pheromones resulted in new state records for seven cerambycid species (Handley et al. 2015). In similar fashion, in a proof of concept study, pheromone-baited traps were used to detect and assess numbers of the endangered valley elderberry longhorn borer, Desmocerus californicus dimorphus. This single study in one season using pheromone-baited traps caught (and released) more beetles than the total in all collections in the US. We also identified pheromones for three dangerous invasive species which are already present in the US or have been intercepted in shipments entering the US, including Callidiellum rufipenne, Anoplophora chinensis, and Xylotrechus rufipenne. With the help of collaborators, we are expanding these studies to Australia, South America, and Africa, with a particular focus on invasive species. Objective 2.To proactively identify semiochemicals of other exotic pests that are likely to be introduced into the US through global commerce, and develop lures that can be used for their detection. We initiated a study on the sex attractant pheromones of North American wireworm species. Although these species are endemic rather than invasive, they are rapidly becoming significant problems in many crop systems due to changes in insecticide use. No pheromones are known from any North American species; two reports of pheromone identifications from several decades ago have proven to be incorrect. As part of the work, we synthesized a small library of 5 pheromones known from European and Asian species, and in field tests, this library has already attracted one species. These trials are being expanded to synthesize and test all known wireworm pheromones, as well as targeting specific species for pheromone identification. The work is also being expanded to include collaborators in Montana, Illinois, and South Carolina, who will both field test the pheromones we make, and prepare pheromone gland extracts from their local species. Objective 3: To explore the role of contact pheromones as mediators of insect behavior, and where feasible, develop methods for exploiting those signals for insect management. This work included a fundamental study to assess the chirality of methyl-branched hydrocarbons found in the cuticular lipids of numerous insects, many of which function as contact chemical signals for recognition of species and sex, and in social insects, for nestmate recognition, caste labeling, and queen pheromones. We found that 36 species in 9 different orders without exception produced methyl-branched hydrocarbons with the (R)-configuration, showing that the biosyntheses of these important signals is ancient and highly conserved. This knowledge will be crucial for the identification of the active pheromone blends, particularly for social insects, in which these compounds play a crucial role as queen pheromones. Thus, it may be possible to disrupt colony organization and reproduction by manipulation of these signals. Developing the methodology to isolate, identify, and synthesize the compounds has led to the development of several ongoing collaborations with groups working with ants, wasps, bees, and termites. Objective 4.To conduct preliminary studies on newly discovered invasive species, both to get a head start on developing management programs, and to obtain preliminary data to drive grant proposals. During the grant period, we conducted pilot studies investigating the feasibility of developing attractants for invasive snail and slug species. This provided the preliminary data required to drive a federal grant proposal, which was subsequently funded.

Publications

• Type: Journal Articles Status: Published Year Published: 2015 Citation: K. Handley, J. Hough-Goldstein, L. M. Hanks, J. G. Millar, and V. DAmico. 2015. Species Richness and Phenology of Cerambycid Beetles in Urban Forest Fragments of Northern Delaware. Annals of the Entomological Society of America. Ann. Entomol. Soc. Am. 108:251-262.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Robert F. Mitchell1 , Peter F. Reagel1!, Joseph C. H. Wong1, Linnea R. Meier1, Weliton Dias Silva3, Judith Mongold-Diers1, Jocelyn G. Millar2, and Lawrence M. Hanks. 2015. Cerambycid beetle species with similar pheromones are segregated by phenology and minor pheromone components. Journal of Chemical Ecology 41: 431-440
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Yutaka Narai,1* Yunfan Zou2, Kyoshi Nakamuta,3 Judy A. Mongold-Diers,4 Lawrence M. Hanks,4 and Jocelyn G. Millar2. 2015. Candidate aggregation pheromones of two potentially invasive Asian cerambycid species in the genus Xylotrechus. Journal of Economic Entomology, 108(3):1444-1446
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Yunfan Zou, Jocelyn G. Millar, J. Scott Blackwood, Ryan Van Duzor, Lawrence M. Hanks, Judith A. Mongold-Diers, Joseph C.H. Wong, and Ann M. Ray. 2015. (E)-2-hydroxyoct-4-en-3-one, a male-produced attractant pheromone of the cerambycid beetle Tylonotus bimaculatus. Journal of Chemical Ecology 41: 670-677.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Ann M. Ray, Jocelyn G. Millar, Jardel A. Moreira, J. Steven McElfresh, Robert F. Mitchell, James D. Barbour, and Lawrence M. Hanks. 2015. North American species of cerambycid beetles in the genus Neoclytus share a common hydroxyhexanone-hexanediol pheromone motif. Journal of Economic Entomology 108: 1860-1868
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Laura Hansen, Xu Tian, Jacob Wickham, Yi Chen, Lawrence M. Hanks, Jocelyn G. Millar, Stephen A. Teale. 2015. Identification of a male-produced pheromone component of the citrus longhorned beetle, Anoplophora chinensis. PLoS One. 10(8), e0134358
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Yunfan Zou, Claire E. Rutledge, Kiyoshi Nakamuta, Chris T. Maier, Lawrence M. Hanks, Austin B. Richards, Emerson S. Lacey, and Jocelyn G. Millar. 2016. Identification of a Pheromone Component and a Critical Synergist for the Invasive Beetle Callidiellum rufipenne (Coleoptera: Cerambycidae). Environmental Entomology 45:216-222
• Type: Book Chapters Status: Awaiting Publication Year Published: 2016 Citation: John C. Palumbo, Thomas M. Perring, Jocelyn G. Millar and Darcy A. Reed. 2015. Biology, Ecology, and Management of an Invasive Stink Bug, Bagrada hilaris, in North America. Annual Review of Entomology. Submitted April 2015
• Type: Book Chapters Status: Awaiting Publication Year Published: 2016 Citation: Donald C. Weber, Ashot Khrimian, Maria C. Blassioli-Moreas, and Jocelyn G. Millar. 2016. Semiochemistry of Pentatomoidea, Chapter 14 in Biology of the Pentomoidea, Stink Bugs and Their Relatives. Eds. Jay McPherson and Thomas Miller. Taylor and Francis
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Jan E. Bello, J. Steven McElfresh, and Jocelyn G. Millar. 2015. Isolation and determination of absolute configurations of insect-produced methyl-branched hydrocarbons. Proceedings of the National Academy of Sciences of the USA 112:1077-1082
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Wheeler, C.A., Millar, J.G. and Card�, R.T., 2015. Multimodal signal interactions in the ladybeetle, Hippodamia convergens, aposematic system. Chemoecology, 25:123-133

Progress 09/01/14 to 09/30/14

Outputs
Target Audience: Target audiences served by the project include: Academics and research scientists working in chemical ecology and entomology, with an emphasis of applying chemical ecology to protection of crops and natural habitats Regulatory officials working in local, state, national, and international agencies charged with crop protection Agricultural industry/commodity boards and related grower groups. The general public through nontechnical publications and presentations. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Under the auspices of this project, 2 PhD students in Entomology and Postdoctoral level scientist are being trained. How have the results been disseminated to communities of interest? Please note that this report covers only a one month period from Sept 1 to Sept 30, 2014. No papers were published nor presentations made during this one month period What do you plan to do during the next reporting period to accomplish the goals? The project has three main focii, and work is progressing on all three, as follows: Focus 1) identification and exploitation of semiochemical signals used by native and exotic cerambycid beetles. Since the start of the project last September, we have identified and synthesized the first examples of two new pheromone motifs for cerambycids. We have started testing one fo these compounds in southern California, where the early beetle species are flying already. Later in the season, these compounds will be tested in field trials in northern California, the Midwest, and China, with the help of collaborators. 2) "proactive chemical ecology", loosely defined as the identification of pheromones and related compounds that can be exploited for detection and control of invasive insect pests that are not yet present or not yet firmly established in the US. We are currently working on the synthesis of some trial attractants for Giant African Land Snail, an invasive species which has become established in limited areas of Florida, where it is the subject of an intensive eradication effort, and in Hawaii. The snail is not yet established in California, but it has been intercepted in incoming shipments a number of times, so a sensitive detection method based on attractants would be immediately useful to regulatory agencies. We have also been carrying out preliminary experiments on possible repellents for polyphagous shothole borer, an invasive beetle species that has started to cause extensive damage in avocadoes and numerous ornamental tree species in California. This pest invaded Israel several years ago, where it has caused major damage to their avocado industry. 3) A detailed examination of the roles of contact pheromones as mediators of insect behavior, particularly for social insects, and manipulation of these signals for insect management. We recently published a manuscript describing methods for isolating individual cuticular hydrocarbons from insects, so that the functional roles of individual compounds as fertility and dominance signals in social insect colonies can be determined. This may provide new leads for control of pest species by disruption of their colony structure. THis has resulted in collaborations with 6 collaborations so far, in which my group will focus on the chemistry of the bioactive compounds, and our collaborators will work out the behaviors that are mediated by specific compounds that we have identified and as needed, synthesized. At the level of publication and presentation that my group has maintained over the past several years (51 papers published since 2011, with 3 currently in press and another 6 submitted for review; 60 invited papers as presenter or coauthor since 2011), I anticipate that during the course of the project, we will publish a minimum of 6 papers a year associated with the project, with a similar number of presentations at conferences or other public forums

Impacts
What was accomplished under these goals? Please note that this report covers only a one month period from Sept 1 to Sept 30, 2014.

Publications