2005 Annual Report 1.What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter? Introduction of exotic invasive species that become pests pose an increasing threat to U.S. agriculture, urban landscapes, forests, human health and the environment. The Asian Longhorned Beetle (ALB), a wood borer, is among the high risk invasive species that recently invaded the U.S. from China. In China, ALB is considered one of the five most important tree pests. It is reported to attack over 100 deciduous tree species and to have killed hundreds of millions of trees across 25 provinces. Its geographic range extends across climatic zones in China that correspond to North American regions from southern Mexico to southern Canada, including the Maritime Provinces and British Columbia. Cargo infested with ALB has been discovered and eliminated in warehouses in at least 17 states across the U.S., as well as in Canada, Italy, Austria, Germany, France and Poland. To date, known infestations outside its countries of origin (year found) include New York City and Long Island, NY (1996), Chicago, IL (1998), Braunau, Austria (2001), Jersey City, NJ (2002), Gien, France (2003), Toronto, Canada (2003) and Carteret, New Jersey (2004). Most recently, adult ALB were discovered in Sacramento, CA (June 2005), putting at risk many tree species in western U.S. In North America (U.S. and Canada), ALB has attacked at least 25 deciduous tree species in 13 genera, most notable seven maple species. ALB appears to prefer to attack healthy trees. To date (September 2005), over 17,000 (7,190 in NY; 1773 in IL; 8,726 in NJ) and 23,000 of these high value shade trees have been cut and removed in the U.S. and Canada, respectively, in an effort to eradicate ALB and prevent its permanent establishment. However, adult beetles and/or newly infested trees continue to be found in both the NY and NJ infestations. In 2004-05, at least five new areas were found infested in New York City, including Central Park in Manhattan. Although presently limited in distribution within the U.S. and Canada, its potential for spread into other urban centers, as well as into North American watersheds and forests is alarming and demands immediate attention. A recent report projects that if ALB spreads to urban trees across the U.S., there could be a loss of 35% of total canopy cover (1.2 billion trees), at a compensatory value loss of $669 billion. Furthermore, for the lumber, maple syrup, tourist and other forest-related industries, survey, detection and management costs and lost revenues could mount into the billions of dollars. In addition, structural weakening of trees by larvae feeding within infested trees also poses a physical danger to pedestrians and vehicles from falling limbs or trees. In regard to control, systemic insecticides applied by trunk injection and soil injection are currently used in the U.S. However, since efficacy of insecticides depends upon the exposure of ALB to a lethal dose, the efficacy of systemic insecticides is of concern for at least four key reasons. (1) The uptake of imidacloprid, the insecticide of choice, within trees is not uniform. This results in: (a) highly variable distribution and residual levels of imidacloprid within and between branches within treated trees; and (b) highly variable residual levels of imidacloprid within treated trees over time. (2) There is evidence that imidacloprid, in its current formulation and injection method(s), and the resulting levels within treated trees, causes both ALB larvae and adult beetles to stop feeding. Evidence showed that many of the adult beetles that do feed on treated trees recover and are not killed by the small amount of insecticide ingested (similar evidence exists for other wood borers). Therefore, these results suggest that injecting imidacloprid into a buffer zone of trees may not form an impenetrable barrier of trees intended to kill and halt the spread of ALB. On the contrary, while imidacloprid may kill some larvae and adult ALB, other ALB simply stop feeding, recover, spread and continue to feed and infest new trees. (3) While there is evidence that imidacloprid is more effective at killing adult ALB while they feed on twigs and leaves of poplar trees than other tested insecticides, its efficacy (% control of adult population) and its ability to protect trees from attack have not been tested. (4) Finally, there is antidotal evidence that prophylactic injection of imidacloprid is not 100% effective since newly attacked trees that had been repeatedly injected with imidacloprid over several years were subsequently found infested in the New York infestation. Therefore, the only method thus far proven to control ALB is the identification and removal of infested trees. However, this too is problematic since identification of infested trees, which is limited to visual inspection, is at best 60% effective (APHIS studies). Therefore, methods for survey, detect and control of ALB must be developed. Since ALB threatens our urban, suburban, forest and other natural landscapes, including important watersheds, the work outlined in this CRIS is relevant to the U.S., Canada and Europe, as well as China and Korea; to forest products and forest-related industries; and to the maple syrup and tourism industries. The first aim of this project involves intensive complementary investigations, in its countries of origin and invasion (e.g. U.S., Canada), of the biology, behavior and ecology of the Asian Longhorned Beetle (ALB), with the following objectives: (1) to identify ecological and behavioral traits vulnerable to intervention; (2) to develop technologies for control of ALB; (3) to develop detection, monitoring, attract-and-kill strategies for adult ALB; (4) to develop technologies for detection of trees infested by ALB (e.g. acoustic detection); (5) to develop predictive spatiotemporal models (e.g. population spread, seasonal occurrence, population abundance and host colonization) for optimizing implementation and effectiveness of these IPM strategies; (6) to develop unique insights into this cerambycid species that can serve as a template for this poorly understood group of important insect pests; and (7) within the context of invasion biology, to study the process of invasion of ALB in its countries of origin (China and Korea) and invasion (U.S. and Canada), and develop proactive approaches for early detection and rapid eradication of this and other invasive species, thereby reducing the risk of establishment, proliferation and geographic spread of future introductions. The second aim of this project focuses on biological control of the Asian Longhorned Beetle (ALB), with the following objectives: (1) to identify promising candidate parasitoid species within their countries of origin based upon bio-ecological studies, and to evaluate their target and non-target impacts in quarantine; and (2) to identify native North American natural enemies found parasitizing: (a) ALB within North American infestations; and (b) native cerambycid species most closely related to ALB and/or within ALB hosts. For the most promising candidate species, we will: (1) develop mass rearing technology; (2) develop protocols for inundative releases or introduction; and (3) evaluate their impact on ALB and non-target species. This project involves cooperation with Simon Fraser University for GC-EAD analysis of host attractants and development of technology for detection of adult ALB; with Institute of Zoology (Chinese Academy of Sciences) for field evaluation of host attractants; with State University of New York, Pryor Knowledge Systems, Inc. and ARS Bioacoustics Lab for acoustical analysis and development of acoustic technology for detection of ALB larvae feeding inside infested trees; with Sino-American Biological Control Lab (Chinese Academy of Agricultural Sciences) for development of alternative methods of control and attract-and-kill strategies for adult ALB; with Department of Entomology (Cornell University) for development of fungal pathogens for biological control of ALB; with Canadian Forest Service and the Canadian Food Inspection Agency for census of the ALB infestation in Toronto, coupled with Department of Evolutionary Biology (Cornell University) for molecular analysis of ALB, which collectively will result in development of spatiotemporal models of the process of invasion of a founder population of this invasive species; with Chinese Academy of Forestry for biological control of ALB using natural enemies native to China; and with the Departments of Entomology (University of Illinois and University of Vermont) for biological control of ALB using natural enemies native to the U.S. Complementary field studies conducted within the countries of origin and invasion will analyze the processes of invasion, and clarify the mechanisms and predictors of invasion at the molecular, biological, behavioral, and ecological levels. This will include studies in a variety of landscapes at both the individual and population levels. The potential impact of the work outlined in this CRIS will be the development of technologies where none exist, including: (1) development of survey, detection and control methods for ALB; (2) development of biological control; (3) identification of resistant hosts and development of replanting and reforestation guidelines; and (4) development of predictive spatiotemporal models of ALB emergence and dispersal, and population abundance and spread that will collectively optimize the implementation of these strategies, as well as provide methods for evaluating their effectiveness. Furthermore, these studies will result in: (1) the identification and development of predictors of potentially invasive species; (2) proactive development of detection, eradication and management technologies for those species at greatest risk of invasion and establishment; (3) reduced reliance upon insecticide by developing alternative approaches that are ecologically, economically, and socially acceptable; and (4) managing and safeguarding resources from exotic insect pests based on sound technologies. Finally, in anticipation of possible failure to eradicate ALB, and with the continuous threat of re-establishment in these and other regions, the potential impact of the work outlined in this CRIS is to develop new environmentally sound technologies that will form the framework for management. Three categories of customers will benefit from results of this project. The first is users of published research, who are scientists at universities, state departments of agriculture and natural resources, federal agencies, and conservation organizations, and decision-makers at state departments of agriculture and natural resources, federal agencies, conservation organizations. The second includes land and resource managers who are responsible for trees attacked by the target pests, and who benefit from reduced production costs and reduced exposure to toxic substances. The third category of customers is the general public, who benefit from reduced health risks from pollution (e.g. air pollution); who benefit from the water purifying action of trees grown within watersheds (e.g. a $9 billion water treatment plant would be required to replace the Catskill water shed that provides over 70% of the drinking water for New York City); and who benefit from reduced prices, and reduced impact on non-target species. This research falls under National Program 304, `Crop Protection and Quarantine', and addresses Component II ‘Biology of pests and natural enemies’, including Sections A. ‘Basic Biology’ and B. ‘Rearing Insects and Mites’ (natural enemies for inundative and augmentative biological control); Component IV ‘Post Harvest, Pest Exclusion, and Quarantine Treatment’, including Sections A. ‘Detection and Delimitation of Exotic Insect Pests’, C. ‘Control and Eradication of Exotic Insect Pests and D. Fundamental Biology and Ecology of Exotic Insect Pests’; and Component V ‘Pest Control Technologies’, including Sections A. ‘Traditional Biological Control’ and D. ‘Other Biologically-Based Control’. 2.List the milestones (indicators of progress) from your Project Plan. 2006 Objective 1. Biology, Behavior and Ecology of the Asian Longhorned Beetle (ALB) 1.1. Host Selection and Host Preference: 1.1.1. Sentinel Trees (detection of adult ALB) 1.1.1.1. Begin evaluation of attraction among Chinese and North American tree species (Acer mono vs. A. truncatum) - 2006 1.1.1.2. Begin evaluation of sentinel tree plus contact insecticide plus trap (Demand repellency) – 2006 1.1.1.3. Begin evaluation of insecticide bands plus trap – 2006 1.1.2. Artificial Lures (detection of adult ALB) 1.1.2.1. Begin GC-EAD analysis of attractive sentinel tree volatiles – 2006 1.1.2.2. Begin olfactometer evaluation of host odors – 2006 1.1.2.3. Begin field evaluation of host odors (e.g. formulation, release rates) – 2006 1.2. Host Colonization and Suitability 1.2.1. Continue comparative host colonization behavior – 2006 1.3. Detection Technologies for ALB-Infested Hosts 1.3.1. Develop acoustic technology 1.3.1.1. Evaluate current sensor sensitivity - 2006 1.3.2. Develop improved field guide for visual detection of ALB infested trees – 2006 1.3.3. Develop genetic marker-based diagnostic tools for ALB-infested trees 1.3.3.1. Identify expressed sequence tags (ESTs) from ALB larvae - 2006 1.3.3.2. Identify putative full-length proteins from ALB larvae – 2006 1.3.3.3. Begin evaluation of gene expression from non-infested and infested maples to identify genes linked to infestation by ALB – 2006 1.4. Population Suppression and Eradication Technology 1.4.1. Contact insecticide 1.4.1.1. Continue evaluation of insecticide spray – 2006 1.4.2. Fungal bands 1.4.2.1. Begin evaluation of efficacy as a function of population density – 2006 1.5. Processes and Mechanisms of Invasion Biology 1.5.1. Census of ALB infestation in Toronto 1.5.1.1. Continue collection of ALB infestation data – 2006 1.5.1.2. Begin ALB infestation data analysis – 2006 1.5.2. Specimen Collection and Molecular Analysis 1.5.2.1. Continue specimen colleciton ; Begin Analysis of Indigenous Geographic-Populations (China) - 2006 1.5.2.2. Continue specimen collection ; Begin Analysis of Indigenous Host-Populations (China) – 2006 Objective 2. Biological Control for the Asian Longhorned Beetle (ALB). 2.1. Locate suitable investigation areas in Far East 2.1.1. Renew/establish SCA’s needed in Asia. Generate CLIMEX overlays - 2006 2.2. Assessment of natural enemy complex 2.2.1. Begin inventory, obtain identifications - 2006 2.3. Role of natural enemies in ALB population dynamics 2.3.1. Test methodologies and equipment - 2006 2.4. Prioritization and importation of natural enemies 2.4.1. Assess information on known ALB natural enemies - 2006 2.5. Host specificity tests on candidate natural enemy species 2.5.1. Develop list of non-target species for testing - 2006 2.6. Studies of native parasitoids of longhorn beetles 2.6.1. Complete literature search and parasite-host catalog. Begin surveys. - 2006 2.6.2. Continue surveys. Begin trials with ALB. – 2006 2007 Objective 1. Biology, Behavior and Ecology of the Asian Longhorned Beetle (ALB) 1.1. Host Selection and Host Preference: 1.1.1. Sentinel Trees (detection of adult ALB) 1.1.1.1. Continue evaluation of attraction among Chinese and North American tree species (A. mono vs. Eleagnus vs. Tilia) - 2007 1.1.1.2. Complete evaluation of sentinel tree plus contact insecticide plus trap (Demand repellency) – 2007 1.1.1.3. Complete evaluation of insecticide bands plus trap – 2007 1.1.2. Artificial Lures (detection of adult ALB) 1.1.2.1. Continue GC-EAD analysis of attractive sentinel tree volatiles – 2007 1.1.2.2. Continue olfactometer evaluation of host odors - 2007 1.1.2.3. Continue field evaluation of host odors (e.g. formulation, release rates) –2007 1.2. Host Colonization and Suitability 1.2.1. Complete comparative host colonization behavior – 2007 1.3. Detection technologies for ALB-infested hosts 1.3.1. Develop acoustic technology 1.3.1.2. Begin evaluation of current neural network system in the field - 2007 1.3.1.3. Evaluate different sensors - 2007 1.3.3. Develop genetic marker-based diagnostic tools for ALB-infested trees 1.3.3.3. Continue evaluation of gene expression from non-infested and infested maples to identify genes linked to infestation by ALB – 2007 1.4. Population suppression and eradication technology 1.4.1. Contact insecticide 1.4.1.1. Complete evaluation of insecticide spray – 2007 1.4.2. Fungal bands 1.4.2.1. Complete evaluation of efficacy as a function of population density – 2007 1.4.2.2. Begin evaluation of efficacy as a function of band placement and density – 2007 1.5. Processes and Mechanisms of Invasion Biology 1.5.1. Census of ALB infestation in Toronto 1.5.1.1. Complete collection of ALB infestation data – 2007 1.5.1.2. Continue ALB infestation data analysis – 2007 1.5.2. Specimen Collection and Molecular Analysis 1.5.2.1. Complete specimen colleciton ; Continue Analysis of Indigenous Geographic-Populations (China) - 2007 1.5.2.2. Complete specimen collection ; Continue Analysis if Indigenous Host-Populations (China) - 2007 1.5.2.3. Specimen collection; Begin Analysis of Expanded Populations (China) – 2007 1.5.2.4. Begin Analysis of Founder Population (Canada) – 2007 Objective 2. Biological Control for the Asian Longhorned Beetle (ALB). 2.1. Locate suitable investigation areas in Far East 2.1.2. Begin sampling primary and secondary sites - 2007 2.2. Assessment of natural enemy complex 2.2.2. Continue inventory, obtain identifications - 2007 2.3. Role of natural enemies in ALB population dynamics 2.3.2. Collect data at study sites - 2007 2.4. Prioritization and importation of natural enemies 2.4.2. Begin biological studies on imported species - 2007 2.5. Host specificity tests on candidate natural enemy species 2.5.2. Life history and host range studies on promising candidate species - 2007 2.6. Studies of native parasitoids of longhorn beetles 2.6.3. Continue surveys. Continue trials with ALB – 2007 2008 Objective 1. Biology, Behavior and Ecology of the Asian Longhorned Beetle (ALB) 1.1. Host Selection and Host Preference: 1.1.1. Sentinel Trees (detection of adult ALB) 1.1.1.1. Complete evaluation of attraction among Chinese and North American tree species (A. platanoides vs. A. saccharum vs. A. saccharinum vs. A. rubrum) - 2008 1.1.1.4. Begin evaluation of attraction among different ALB-infested tree species – 2008 1.1.2. Artificial Lures (detection of adult ALB) 1.1.2.1. Complete GC-EAD analysis of attractive sentinel tree volatiles – 2008 1.1.2.2. Complete olfactometer evaluation of host odors - 2008 1.1.2.3. Continue evaluation of host odors (e.g. formulation, release rates) – 2008 1.3. Detection technologies for ALB-infested hosts 1.3.1. Develop acoustic technology 1.3.1.2. Complete evaluation of current neural network system in the field - 2008 1.3.1.4. Begin evaluation of neural network and optimal sensors – 2008 1.3.3. Develop genetic marker-based diagnostic tools for ALB-infested trees 1.3.3.3. Compete evaluation of gene expression from non-infested and infested maples to identify genes linked to infestation by ALB – 2008 1.3.3.4. Begin development of these genetic markers into a diagnostic method for the early detection of ALB infestations in maple – 2008 1.4. Population suppression and eradication technology 1.4.2. Fungal bands 1.4.2.2. Complete evaluation of efficacy as a function of band placement and density – 2008 1.5. Processes and Mechanisms of Invasion Biology 1.5.1. Census of ALB infestation in Toronto 1.5.1.2. Continue ALB infestation data analysis – 2008 1.5.2. Specimen Collection and Molecular Analysis 1.5.2.1. Complete Analysis of Indigenous Geographic-Populations (China) - 2008 1.5.2.2. Complete Analysis if Indigenous Host-Populations (China) - 2008 1.5.2.3. Continue Analysis of Expanded Populations (China) – 2008 1.5.2.4. Continue Analysis of Founder Population (Canada) – 2008 Objective 2. Biological Control for the Asian Longhorned Beetle (ALB). 2.1. Locate suitable investigation areas in Far East 2.1.3. Finish sampling primary and secondary sites. Analyze parasite complexes - 2008 2.2. Assessment of natural enemy complex 2.2.3. Complete inventory, summarize findings – 2008 2.3. Role of natural enemies in ALB population dynamics 2.3.3. Collect data at study sites. Analyze data - 2008 2.4. Prioritization and importation of natural enemies 2.4.3. Continue biological studies on imported species - 2008 2.5. Host specificity tests on candidate natural enemy species 2.5.3. Begin host range studies on promising candidate species - 2008 2.6. Studies of native parasitoids of longhorn beetles 2.6.3. Continue surveys. Continue trials with ALB – 2008 2009 Objective 1. Biology, Behavior and Ecology of the Asian Longhorned Beetle (ALB) 1.1. Host Selection and Host Preference: 1.1.1. Sentinel Trees (detection of adult ALB) 1.1.1.4. Complete evaluation of attraction among different ALB-infested tree species - 2009 1.1.1.5. Begin evaluation of attractive radius of promising sentinel tree species – 2009 1.1.2. Artificial Lures (detection of adult ALB) 1.1.2.3. Complete evaluation of host odors (e.g. formulation, release rates) – 2009 1.1.2.3. Begin evaluation of attractive radius of most promising lures – 2009 1.2. Host Colonization and Suitability 1.2.1. Begin evaluation of host suitability among North American tree species –2009 (Quercus rubrum (northern red oak), Quercus lobata (valley oak); Aesculus california (California buckeye), Olea europaea (common olive); Eucalyptus globulus (Blue gum eucalyptus); Platanus racemosa (California Plane), Platanus rientalis (Oriental Plane) 1.3. Detection technologies for ALB-infested hosts 1.3.1. Develop acoustic technology 1.3.1.4. Continue evaluation of neural network and optimal sensors – 2009 1.3.3. Develop genetic marker-based diagnostic tools for ALB-infested trees 1.3.3.4. Continue development of genetic markers into a diagnostic method for the early detection of ALB infestations in maple – 2009 1.5. Processes and Mechanisms of Invasion Biology 1.5.1. Census of ALB infestation in Toronto 1.5.1.2. Continue ALB infestation data analysis – 2009 1.5.2. Specimen Collection and Molecular Analysis 1.5.2.3. Complete Analysis of Expanded Populations (China) – 2009 1.5.2.4. Complete Analysis of Founder Population (Canada) – 2009 Objective 2. Biological Control for the Asian Longhorned Beetle (ALB). 2.4. Prioritization and importation of natural enemies 2.4.3. Complete biological studies on imported species - 2009 2.5. Host specificity tests on candidate natural enemy species 2.5.3. Complete host range studies on promising candidate species - 2009 2.6. Studies of native parasitoids of longhorn beetles 2.6.3. Complete surveys. Complete trials with ALB – 2009 2010 Objective 1. Biology, Behavior and Ecology of the Asian Longhorned Beetle (ALB) 1.1. Host Selection and Host Preference: 1.1.1. Sentinel Trees (detection of adult ALB) 1.1.1.5. Complete evaluation of attractive radius of promising sentinel tree species – 2010 1.1.2. Artificial Lures (detection of adult ALB) 1.1.2.3. Complete evaluation of attractive radius of most promising lures – 2010 1.2. Host Colonization and Suitability 1.2.1. Continue evaluation of host suitability among North American tree species – 2010 (Quercus rubrum (northern red oak), Quercus lobata (valley oak); Aesculus california (California buckeye), Olea europaea (common olive); Eucalyptus globulus (Blue gum eucalyptus); Platanus racemosa (California Plane), Platanus rientalis (Oriental Plane) 1.3. Detection technologies for ALB-infested hosts 1.3.1. Develop acoustic technology 1.3.1.4. Complete evaluation of neural network and optimal sensors – 2010 1.3.3. Develop genetic marker-based diagnostic tools for ALB-infested trees 1.3.3.4. Complete development of genetic markers into a diagnostic method for the early detection of ALB infestations in maple – 2010 1.5. Processes and Mechanisms of Invasion Biology 1.5.1. Census of ALB infestation in Toronto 1.5.1.2. Complete ALB infestation data analysis – 2010 Objective 2. Biological Control for the Asian Longhorned Beetle (ALB). 2.6. Studies of native parasitoids of longhorn beetles 2.6.4. Rank most promising North American species found - 2010 4a.What was the single most significant accomplishment this past year? This project superseded project number 1926 22000 012 00D, terminated on 07/06/2005, has only been in place two months. See the final report for that CRIS for a description of significant accomplishments for FY 2005. 4b.List other significant accomplishments, if any. None 4c.List any significant activities that support special target populations. None 4d.Progress report. 1926-22000-019-01S: This report serves to document research conducted under a specific cooperative agreement between ARS and Chinese Academy of Forestry, Beijing, CHINA. Investigations on Scleroderma guani and Dastarcus longulus, two natural enemies of ALB in China, were continued. We continue to desire the importation of these two natural enemies to BIIRL for their evaluation on non-target effects, as well as mass rearing and inundative release for control of ALB should non-target effects prove to be acceptable. Assistance from ARS Sino-American Biological Control Laboratory, Beijing for importation is necessary. 1926-22000-019-02S: This report serves to document research conducted under a specific cooperative agreement between ARS and the State University of New York (Syracuse). The objectives of the SCA were to continue development of the Asian Longhorned beetle acoustic detection system and to continue the identification of oviposition deterrents. The acoustic detection system has progressed to the point where a prototype field unit was produced and will be tested in the field by APHIS in 2005. Evaluation of the sensitivity of the existing sensor, as well as several other sensors, will be conducted in 2005 in collaboration with the ARS Center for Medical, Agricultural, and Veterinary Entomology lab, Gainesville, Florida. Analysis of acoustical signatures of ALB feeding with three tree species, three host tissues, under three temperature and wood moisture conditions was initiated in cooperation with the ARS Center for Medical, Agricultural, and Veterinary Entomology lab, Gainesville, Florida. Analyses will be completed in 2006. 1926-22000-019-03S: This report serves to document research conducted under a specific cooperative agreement between ARS and the University of Illinois at Urbana-Champaign, Urbana, IL. Progress Item 1a. Review of Literature - We have combined into a single document three EndNote reference lists of papers concerning parasitoids of cerambycid beetles, including lists originally compiled by M. Smith, R. Fuester, and L. Hanks. We are continuing to collect new references and add them to the EndNote database. The summary of these data have been presented in the symposium Variations on a theme-Biological Control at the North-Central Branch of the Entomological Society of America, and at National Meetings of the ESA (2004), Updated results will be presented later this year at the IX European Workshop on Insect Parasitoids, Cardiff Univ., Wales (September 2005) and at the National ESA meetings, Fort Lauderdale, FL (November 2005). Progress Item 1b. Identify Native Natural Enemies of Native Wood Borers in Illinois and Surrounding Areas - We summarize here the results obtained from trees felled during 2003-2004. We collected downed wood of 12 tree species, and caged the material to rear wood-boring beetles and their parasitoids. We reared 9 cerambycid species, 24 species of parasitic hymenoptera (primarily ichneumonids and braconids), and 1 species each of phorid and tachinid flies. The most common parasitoid species is Wroughtonia ferruginea, a polyphagous species that attacked at least three cerambycid species in our study. During 2005, we felled 6 Norway maples and pin oaks, and girdled and half-girdled another 6 trees per species. We also felled three white pines per the project plan. Logs of all study trees were later enclosed in fine mesh material to capture emerging egg parasitoids. This fall, we have begun enclosing other logs in aluminum screening to capture emerging beetles and parasitoids. To date, we have captured adults of one species of cerambycid on felled maple trees, two species on felled pin oaks, and 1 species on the pines. We also have captured adults of two species of braconid parasitoids on the felled maples and one species on the pin oaks. We have not yet seen any adult cerambycids or parasitoids on the girdled or half-girdled trees, although the former clearly are showing signs of severe stress (producing epicormic shoots, leaves wilting). 2.6. Studies of native parasitoids of North American longhorn beetles: During 2005, we stressed 4 tree species: red maple (Acer rubrum), pignut hickory (Carya glabra), mockernut hickory (Carya tomentosa), and Virginia pine (Pinus virginiana). Trees were stressed by one of three methods: half-girdling, complete-girdling and felling. Trees were inspected on a weekly basis to identify if and when colonization by attacking insects occurred. Upon signs of colonization, bolts were cut and enclosed in fine mesh material to capture emerging egg parasitoids, or in aluminum screening to allow entry of natural enemies. Bolts enclosed in aluminum screening will be held as such until June 2006, at which time they will be enclosed in fine mesh material to capture emerging beetles and parasitoids. To date, we observed signs of colonization on the felled Virginia pine. Similar studies, under a new Specific Cooperative Agreement with the University of Vermont, were initiated using sugar maple (Acer saccharum) and white pine (Pinus strobus). In addition, ALB-infested bolts from the Carteret, NJ infestation were reared at BIIRL and emerging natural enemies collected. To date, numerous braconid parasitoids has been collected and awaiting taxonomic identification. These results tentatively indicate that natural enemies native to the U.S. parasitize ALB in nature. Finally, we have also been collecting parasitoids from sites with logging debris containing roundheaded and flatheader borers in Pennsylvania State Game Lands. Two approaches have been used, capturing ovipositing females in the field and obtaining emergence from bolts collected and placed in an outdoor insectary. So far, we have obtained an Atanycolus sp. (Braconidae) in abundance. These have been exposed to logs containing ALB in quarantine, but no successful parasitism has been observed as yet. In addition, we have recovered two ichneumonid species which have undergone limited testing, with no parasitism as yet. Finally, we have recovered a tachinid species (two females) and a chalcidoid species (one female), which we will test if any males emerge. All of the state game lands in SE Pennsylvania undergo a clearcut rotation, so fresh material should be available throughout the growing season for years to come. 1926-22000-019-04S: This report serves to document research conducted under a specific cooperative agreement between ARS and Cornell University, Ithaca, New York, USA. The project objective is to develop methods for optimizing control of Asian Longhorned beetle using entomopathogenic fungi, especially focusing on the use of non-woven fiber bands for fungal delivery. 1.4.3. Fungal bands - FIELD STUDIES: During the 2005 field season we finally conducted a study including a treatment of fungal bands plus attractants in a location in China with a good beetle population. Studies were conducted in a planting of poplar trees (Populus simonii x Populus canadensis) in Friendship Village, northeast of Linhe (N 40o 43-44 x E107o 17-18; altitude = 1046 m), Inner Mongolia, People's Republic of China. Fungal bands were produced by the Anhui Agricultural University, who collaborated throughout this project. The fungal isolate used was Metarhizium anisopliae F-52, which has been registered with the EPA for use for control of ticks and lawn grubs in the US. The attractant that was used had two parts: a contact pheromone supplied by Dr. Aijun Zhang, USDA, ARS and a host plant attractant made by placing bark from twigs of Acer mono in dimethyl sulfoxide. Five plots of ca. 75-125 trees each (8 m wide containing 3 rows of trees x 100 m long) were created for each of three treatments: fungal bands alone, fungal bands plus attractant and controls with no fungal bands. The trees had been planted approximately 17-18 years before. Plots were separated from each other by 70 m of unbanded trees, which were not included in these studies and acted as a buffer zone between treatments. Bands were hung on 48 trees per plot in early-mid July. These trees were not very similar to trees in the northeastern U.S., because on many trees the canopy was not wide, foliage was very sparse and tops were dead. However, these were the trees available with a pre-existing ALB population. Before bands were hung and every 5 days afterward, up to 7 beetles were collected from each plot. More beetles than this were not collected because we did not want to deplete the population so that we would not have adults to collect over time. At each sampling date, the small bags with host plant extract were also refilled. Beetles were reared at constant temperature (25 C) for up to 70 days after collection and date of death was recorded as well as whether M. anisopliae subsequently grew out of cadavers. To evaluate the effect of fungal bands on oviposition and survival of eggs, in early August, 24 trees were cut down. All beetle exit holes were recorded in each tree but more importantly, new ovipositions were recorded and resulting eggs were maintained on wet filter paper under constant temperature to determine whether they hatched or not. For eggs that did not hatch, outgrowth of M. anisopliae was recorded. As background data, the density of adult beetles was recorded once every 5 days to document phenology and relative densities of ALB in different plots. These data were also used to assist when adults were collected. The experiment was terminated in mid-August. Some preliminary data was taken to study environmental contamination by this fungus from inoculated adults. Bark samples were taken after an inoculated adult was exposed to a fungal band and allowed to walk on tree trunks. Spore densities on bark samples were quantified using dodine plates at different dilutions from the washes from bark samples. At this time, the data from all studies have not yet been totally collected and have not been analyzed although computerization of some of the data will begin shortly. The data that were collected will be part of the M.S. thesis of a Cornell University graduate student who was part of the team collecting the data in Linhe. When this study was planned it was very difficult to find a location in a region that received summer rainfall similar to the northeastern U.S., but also provided a robust population of ALB, so that enough beetles would be present for this study. This is a difficulty in conducting such studies because naturally occurring beetle populations in China must be used. After extensive searching by both Drs. Smith and Li, the best study site that could be found was in a semi-desert region, where rainfall was questionable during the period of the study. The weather in the 2005 study area is not similar to normal weather in northeastern North America. While bands were hanging during this study, some rain fell occasionally but for the first two weeks after bands were hung, it was exceptionally hot and dry. Whether these weather conditions will affect the activity of fungal bands is not known although one could assume that this might have a negative impact. However, at present, it is too early to know the results from this study. LABORATORY STUDIES: (1) Fecundity studies: We completed the third replicate of a study of the effects of maternal exposure to M. anisopliae on (a) mates; (b) egg production; (c) egg hatch and infection; and (d) larval survival and infection. We have terrific data and a paper has been started; (2) Dose response: These studies have only begun. First priority over the past year was to complete the Fecundity studies so adult beetles were committed to that study first. Once we had adult beetles, we did not have good fungal bands and making bands took a few tries. Now, has offered fungal bands for out studies and this seems far preferable, to use bands made in the same facility where they have been produced for purchase by APHIS. Then, a method for quantifying the number of viable conidia had to be learned by the technician helping with this project. Thus far, we conducted one dose response replicate but the results are not very consistent with previous studies and, before continuing, we are going to conduct a complete review of the experimental procedures to understand why the results from this bioassay look so different from previous data. 1.3.3. Develop genetic marker-based diagnostic tools for ALB-infested trees: Collaborative studies with Dr. Wayne Hunter were conducted where by: (1) Identify expressed sequence tags (ESTs) from ALB larvae were identified, and (2) putative full-length proteins from ALB larvae were identified. In addition, studies were conducted to develop procedures for successfully implanting ALB larvae into red maple for development, and for collecting and processing samples from non-infested and infested maple trees. 1.5.2.1. Invasion Biology of ALB: Collection of Indigenous Geographic-Populations (China): This report serves to document research conducted under a specific cooperative agreement (1926-22000-012-07S - Agreement # 58-1926-4-0148F) between ARS and Chinese Academy of Sciences, Beijing, CHINA. The objective of this agreement is to, within the context of invasive biology of invasive species, to develop technologies for prediction, detection, monitoring, management and control of invasive species, with current emphasis on the Asian longhorn beetle (ALB), Anoplophora glabripennis. Efforts are in progress for making the collections from within the indigenous range of ALB in China. 5.Describe the major accomplishments over the life of the project, including their predicted or actual impact. This project superceded project number 1926 22000 012 00D on 7/24/2005 and thus has only been in place two months, but the objectives and research in the two projects are closely related. Some of the major accomplishments in the previous project were: (1) development of a spatiotemporal model for predicting dispersal, that was subsequently used for developing survey guidelines for ALB; (2) development of a degree-day model of seasonal population occurrence of ALB, that has subsequently been used in timing the implementation of survey and control strategies in ALB eradication programs; (3) development of improved guidelines for visual detection of ALB-infested trees that has subsequently been used in survey guidelines in ALB eradication programs; (4) developed the first prototype Acoustic Detector for ALB-infested trees; (5) identification of a highly attractive tree species for use in detection of adult ALB; elucidated the mechanism (host odors) for attraction; and developed a method for enhancing attraction; (6) developed Attract-and-Kill technology for detection of adult ALB via the combination of a highly attractive tree species, a contact insecticide with highly effective knockdown capability and a funnel trap; (7) developed a highly effective contact insecticide for ground application to host trees as a optional method to save infested trees and/or protect trees from attack; and (8) development of fungal bands for suppression of ALB. 6.What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end-user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? This project superceded project number 1926 22000 012 00D on 5/24/2005 and thus has only been in place two months. The following are the highlights of accomplishments Detection of Adult ALB Using Acer mono Sentinel Trees – Results from our 2005 studies of the relative attraction of Acer mono and Acer platanoides (Norway maple) sentinel trees, in which A. mono was shown to be significantly more attractive that Norway maple were presented to the California Department of Food and Agriculture as part of the Science Advisory Panel (SAP) in August 2005. Furthermore, results from these studies are being provided to USDA APHIS, state regulatory agencies (NY, NJ, IL and CA), and to the Canadian Food Inspection Agency and Canadian Forest Service (September 2005). The constraints on the adoption of this technology are associated with the availability of A. mono trees in North America. However, the New York Department of Environmental Conservation began growing large numbers A. mono trees in spring 2005 in preparation of using them in subsequent years for detection of ALB. Other suppliers are currently being sought. Control of Adult ALB Using Demand – Although this study is still in progress, results from our 2005 studies, in which Demand was shown to provide 98%-100% control of the adult population in urban maple trees for at least 50 days, as well as rapid knock-down ability, were presented to the California Department of Food and Agriculture as part of the Science Advisory Panel (SAP) in August 2005. The particular importance of this technology is its utilization: (1) to protect trees from ALB attack; (2) to save trees that have previously been under attack, thereby providing an alternative to tree removal; (3) to detect and/or monitor adult beetles; and (4) to kill and prevent the escape and spread of adult beetles when applied to heavily infested trees prior to cutting and removal. Furthermore, results from these studies are being provided to USDA APHIS, state regulatory agencies (NY, NJ, IL and CA), and to the Canadian Food Inspection Agency and Canadian Forest Service (September 2005). Adoption of this technology requires the acquisition of an emergence use label by USDA APHIS or any agency wishing to incorporate use this product in their survey, detection and eradication programs. However, this product is already registered in the U.S. for control of numerous other insect pests, including those attacking trees, and those walking on indoor (e.g. restaurants) and outdoor surfaces of buildings, Therefore, acquisition of an emergency label does not represent a constraint on its adoption. 7.List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: List your peer reviewed publications below). Smith, M.T. “Impact of Invasive Species on the New York City Watershed, with an Emphasis on the Asian Longhorn Beetle”. Manhattan, NY, September 2005. Reagel, P. F., E. S. Lacey, J. F. Tooker, M. D. Ginzel, M. T. Smith, R. W. Fuester, & L. M. Hanks. IX European Workshop on Insect Parasitoids, Cardiff Univ., Cardiff, Wales, September 2005. Hajek, A.E., J.R. Reilly, T. Dubois, M. T. Smitih, L. Bauer, and Z. Li. 2005. Development of fungal bands to assist in eradication of Asian longhorned beetle, Anoplophora glabripennis in the U.S. Invited symposium presentation, Soc. Invertebr. Pathol., Ann. Mtg., Anchorage, Alaska.