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? Fumigation with methyl bromide or phosphine is the method of choice for postharvest insect control in fresh fruits, dried fruits, and tree nuts. In addition, methyl bromide is used in most quarantine treatments required for access of these products to export markets. Regulatory constraints have reduced the availability and significantly increased the cost of methyl bromide, making the fumigant less attractive for these uses. In addition, the effectiveness of phosphine is threatened by the development in pest populations of resistance to this fumigant. The loss of these fumigants will have a significant impact on commodity quality and trade unless suitable alternatives are developed. We seek to devise non-chemical quarantine and postharvest control strategies for fresh and stored products that reduce losses caused by insects and other arthropods. To meet this goal, we will develop alternative physical and biological treatments, and incorporate these into innovative systems approaches and integrated pest management control strategies. We will also develop insect detection systems that can be used to make informed treatment decisions or remove infested commodity. In addition, we will generate the basic biological and environmental data necessary to model the survival, development, and reproduction of pest and beneficial populations as related to alternative treatments. Insects of concern include classical storage pests such as the Indianmeal moth, field pests such as navel orangeworm and driedfruit beetle and quarantine insects such as codling moth, olive fruit fly and glassy-winged sharp shooter. Each year, the western U.S. produces about 8 million tons of dried fruits, tree nuts, citrus fruits, fresh grapes and olives, worth nearly $5 billion. The U.S. must efficiently produce commodities of high quality to maintain both domestic and export markets. By lowering production costs and improving quality of agricultural products, the U.S. may reduce trade deficits, boost market share, and otherwise enhance agricultural competitiveness. Postharvest insects and other arthropods cause an estimated 96 million dollar loss to dried fruits and nuts each year, and seriously affect our ability to export fresh products to foreign markets because of quarantine restrictions. Use of methyl bromide for most applications is to be phased out in the year 2005. Although quarantine treatments will be exempt from use restrictions, the limited availability of methyl bromide may make the fumigant prohibitively expensive. There is no single economical alternative to methyl bromide for controlling insect pests throughout the entire processing, storage, and marketing system for fresh and dried commodities, suggesting a need for control programs that use multiple alternatives. Consequently, alternative non-chemical treatments and strategies that prevent quarantine insects from entering marketing channels and protect durable commodities from insect infestation need to be developed. This research project is relevant to the objectives of National Program 308 – Methyl Bromide Alternatives, specifically the development of nonchemical alternatives such as natural enemies and physical treatments and physical or chemical detection systems for stored product and quarantine pests. The project is also relevant to the objectives of National Program 304 - Crop Protection and Quarantine, specifically the investigation of pest exclusion and quarantine treatment procedures, including the development of accurate and precise pest detection methodology. Cooperative research projects on physical detection systems, transgenic plants, pheromone mating disruption, non-chemical treatments, temperature management, and insect pathogens are being conducted in cooperation with commodity organizations, private industry, University of California at Davis and Riverside, Washington State University, other ARS laboratories (Yakima, WA; Hilo, HI; Manhattan, KS) and various marketing orders and packing houses/commodity storage facilities. 2.List the milestones (indicators of progress) from your Project Plan. Year 1 (FY 2004) Complete time-temperature studies of olive fruit in bins at low temperatures. Determine upper and lower limits of temperature and humidity for olive fruit fly survival. Conduct first year of laboratory studies on the life history of olive fruit fly, and field studies of biological controls and trapping methods for olive fruit fly. Complete laboratory studies of entomopathogenic nematodes and parasitoids against olive fruit fly. Complete laboratory investigations of quarantine strategies to control olive fruit fly Complete comparison of efficiency of olive fruit fly traps Determine heat tolerance of postharvest dried fruit and nut pests Identify stage and pest species most tolerant to vacuum Describe response of cowpea weevil eggs to commercial cold storage temperatures. Complete low temperature studies for eggs of Indianmeal moth and navel orangeworm. Determine seasonal prevalence and spatial variation of navel orangeworm in commercial orchards during a single crop year. Acquire one year of prevalence data for moth pests in vineyards. Collect first year of field data on navel orangeworm mating disruption and control of nitidulid beetles through mass trapping. Collect and contrast chemical signatures from infested and uninfested walnuts. Obtain purified components of Indianmeal moth sex pheromone, and begin comparison of trap baits. Complete first year of trials with insect probe traps in raisin bins. Complete work on cytoplasmic polyhedrosis virus. Continue evaluations of transgenic walnuts for control of postharvest insects. Begin dose-response studies of a gregarine pathogen of Tribolium beetles. Identify and evaluate multi-host pathogens such as nematodes against postharvest insect pests. Evaluate penetration of granulosis virus and B. thuringiensis in product profiles. Year 2 (FY 2005) Complete low temperature mortality studies with fruit in field bins infested with olive fruit fly. Determine combinations of temperature and humidity that limit development of olive fruit fly Continue laboratory and field investigations of control methods and evaluate cultivation and sanitation practices on olive fruit fly populations. Complete field investigations of quarantine strategies to control olive fruit fly. Determine heat tolerance of stored dried fruit and nut beetle pests. Describe response of moth species to commercial cold storage temperatures. Determine seasonal prevalence and spatial variation of navel orangeworm in commercial orchards during a second crop year. Acquire second year of prevalence data for moth pests in vineyards. Collect second season of navel orangeworm mating disruption data Determine dose and scale in nitidulid beetle mass trapping control method. Correlate chemical signatures with level of insect infestation in stored dried fruits and nuts. Describe airflows in and around dried fruit and nut storages; evaluate Indianmeal moth trap baits in various airflow situations. Complete second season of tests of probe traps in raisin bins; develop methods to artificially infest raisins with raisin moth. Continue assessment of gregarine pathogen against Tribolium beetles. Continue field trials with nematodes and explore for other multi-host pathogens. Continue evaluations of transgenic walnuts for control of postharvest insects. Complete study of granulosis virus and B. thuringiensis penetration into stored products held in bins. Complete small scale tests of parasitoids against stored dried fruit and nut insect pests, document reduction of insect fragments in stored dried fruits and nuts using parasitoids. Year 3 (FY 2006) Continue laboratory and field investigations of control methods and evaluate cultivation and sanitation practices on olive fruit fly populations. Complete field studies of nematodes and parasitoids against olive fruit fly. Complete evaluation of olive fruit fly traps. Develop treatment protocols to control pests of stored tree nuts using vacuum in flexible containers. Begin research using artificially infested raisins to determine effect of moth pests on raisin quality. Complete small plot field trials with nematodes in pistachios. Initiate large field trials with nematodes. Complete assessment of gregarines. Complete assessment of entomopathogen penetration in stored product bins. Continue evaluation of transgenic walnuts for control of postharvest insects. Year 4 (FY 2007) Complete assessment of low temperature control methods of olive fruit fly Identify promising radio frequency treatment protocols for disinfestation of walnuts and demonstrate treatments under field conditions, develop treatment protocols for dried fruit and nuts in field bins at cold storage temperatures. Assess olive fruit fly control practices in California. Evaluate spatial variance and auto-correlation of navel orangeworm prevalence in orchards, and correlate prevalence with product damage. Report results of prevalence data for moth pests in vineyards; complete second year of studies using artificially infested raisins to determine effect of moth pests on raisin quality. Complete evaluation of postharvest quarantine control strategies for olive fruit fly, and begin implementation of developed field control techniques. Report findings in navel orangeworm mating disruption; examine optimal timing for attract-and-kill of nitidulids. Complete low temperature studies for eggs of raisin moth. Begin small scale beetle pathogens trials, and select best application method. Complete work on transgenic walnuts for control of postharvest insects by selecting a high expression genetically modified walnut line. Determine relationship of insect control to penetration of pathogens into product. Complete whole room evaluations of parasitoids to control insect pests of stored dried fruit and nut Release nematodes and parasitoids against olive fruit fly in orchards. Implement olive fruit fly quarantine control procedures. Collect and analyze field samples of commodity volatiles and correlate with infestation rates. Implement new olive fruit fly traps. Assess efficacy of mass male trapping of Indianmeal moth using optimal trap bait in optimal locations. Report experiments of pest activity and movement in stored product; gather data from first year of artificial infestation of raisins with raisin moth. Year 5 (2008) Recommend potential low temperature control methods for olive fruit fly. Complete recommendations for control of olive fruit fly based on expected populations. Assess olive fruit fly trapping procedures. Release nematodes and parasites in orchards and evaluate control of olive fruit fly. Modify olive fruit fly control procedures as necessary and implement orchard sanitation practices that reduce olive fruit fly populations. Demonstrate temperature treatments of stored dried fruit and nuts under field conditions Evaluate use of vacuum treatments for dried fruits. Integrate low temperature storage with other control strategies. Complete prevalence studies of moth pests in vineyards. Complete studies and report effects of artificial infestation of moth pests on raisin quality. Complete evaluation of all pathogen trials against stored product insects and recommend candidates for further development. Make recommendation on genetically modified walnuts to walnut industry. Summarize results of granulosis virus and B. thuringiensis penetration studies and recommend optimum application method. Evaluate removal of parasitoids from dried fruits and nuts during processing. Finish nitidulid mass trapping studies. Finish studies optimizing Indianmeal moth pheromone trapping. Gather second year of data using probe traps in artificially infested raisins. 4a.What was the single most significant accomplishment this past year? Use of Radio Frequency Heat Treatments to Control Postharvest Walnut Pests. In order to maintain vital foreign markets, walnut processors need a rapid alternative to methyl bromide for disinfesting walnuts of insect pests. Scientists at the USDA, ARS, San Joaquin Valley Agricultural Sciences Center, Parlier, CA, along with collaborators at Washington State University, University of California, Davis, and Strayfield Limited, have developed a rapid heat method for inshell walnuts using radio frequency technology. Extensive information generated on the heat sensitivity of the major postharvest insect pests of walnuts was used to devise a treatment protocol that effectively disinfested the walnuts without affecting product quality. The efficacy of the treatment, which has been confirmed in the laboratory, is being tested under commercial conditions in a large walnut processing plant. This treatment would be a rapid, non-chemical and effective alternative to methyl bromide, providing the industry with a way to meet the phytosanitary demands of critical markets. 4b.List other significant accomplishments, if any. Biological Control of Olive Fruit Fly. A biological control program was developed for olive fruit fly in California by the USDA, ARS, San Joaquin Valley Agricultural Sciences Center, Parlier, CA in collaboration with the California Olive Committee, Fresno, CA, and the USDA, APHIS, PPQ, MOSCAMED, Guatemala City, Guatemala. Olive fruit fly was recently introduced into California and has the potential to destroy the nation’s olive crop. The imported parasitoid, Psyttalia cf. concolor, was found to parasitize olive fruit fly in laboratory and field tests, and to survive under different climatic conditions in California where olives are grown. The parasitoid did not attack a beneficial seedhead fly used for biological control of yellow star thistle, but was found to parasitize walnut husk fly, a pest of walnuts. A successful biological control program for olive fruit fly in California would help protect the canned olive and olive oil industry in California valued at $90 million annually. Development of a Laboratory Colony of Olive Fruit Fly. A new laboratory colony of olive fruit fly reared on formulated diet was received from University of California Riverside and reared through several generations at the USDA, ARS, San Joaquin Valley Agricultural Sciences Center, Parlier, CA in collaboration with the California Olive Committee, Fresno, CA. The International Atomic Energy Agency laboratory in Seibersdorf, Austria simultaneous received a portion of the same California colony. Maintenance of olive fruit fly for laboratory investigations has been dependent on the seasonal availability of olives, and is less efficient than rearing on formulated diet. The diet and rearing procedures are currently under modification to enhance productivity. Establishment of a stable laboratory colony on formulated diet will provide a means to expedite research to develop control methods including the sterile insect technique. Control of Navel Orangeworm in Almonds Using Mating Disruption. The utility of mating disruption for the control of navel orangeworm in almonds was examined by researchers at the USDA, ARS, San Joaquin Valley Agricultural Sciences Center, Parlier, CA. The navel orangeworm is the primary insect pest of almonds, requiring extensive use of insecticides during production and fumigation to disinfest product. Studies in almond orchards with historically high navel orangeworm abundance and damage examined whether mating disruption and residual insecticides, used together, achieved greater reduction of damage than either alone. Both the mating disruption and insecticide treatments significantly reduced damage, and there was no difference in damage between the insecticide treatment and the plots treated with both mating disruption and insecticide. Adoption of mating disruption for control of navel orangeworm in almonds would help protect this crop, worth an annual average of $1.4 billion farm value, while reducing the ca. 170,000 acres of almonds treated each year with organophosphate insecticides. Navel Orangeworm Prevalence in California Nut Orchards The navel orangeworm is a key pest of high-value California crops such as almonds and pistachios, and a better understanding of population dynamics and host preference is needed. Unmated female-baited flight traps and oviposition traps were used by scientists at the USDA, ARS, San Joaquin Valley Agricultural Sciences Center, Parlier, CA, to confirm differences in seasonal abundance of navel orangeworm between almonds and pistachios. Although navel orangeworm abundance was higher in pistachios than in almonds for much of the season, damage from navel orangeworm larvae was nonetheless lower in pistachios than in almonds, indicating that pistachios are less susceptible to pistachios than almonds. These data show that an area-wide program which successfully reduces abundance of navel orangeworm in both almonds and pistachios, often planted in close proximity, could reduce financial risk from this pest in both crops. Use of Nematodes to Control Navel Orangeworm in Pistachio Orchards. Pistachio growers attempt to reduce overwintering populations of navel orangeworm, a major pest of tree nut crops, by burying infested mummy nuts, but normal disking methods have been shown to be ineffective. Scientists at the USDA, ARS, San Joaquin Valley Agricultural Sciences Center, Parlier, CA, along with ARS scientists from Yakima Agricultural Research Laboratory in Wapato, WA, and collaborators at Paramount Farming Company and CertisUSA, have shown that field application of the insect pathogenic nematode Steinernema carpocapsa is a more effective method for controlling navel orangeworm in mummy nuts. The nematodes were shown to be effective when applied at economical levels in microsprinkler systems, a method that reduces labor costs and ensures adequate moisture for the nematodes. This non-chemical method of reducing overwintering navel orangeworm populations will improve control of this important pest, and reduce the need for insecticide treatments. Chemical Detection of Insect Infestations. Studies to develop an indirect system for detection of insect infestations for stored walnuts were done by ARS personnel in the USDA, ARS, San Joaquin Valley Agricultural Sciences Center, Parlier, CA. Insect infestations are a major cause for customer returns of postharvest walnuts and are difficult to detect. Collections of volatiles from walnuts and insect diet with varying levels of Indianmeal moth infestation were made. Volatiles were extracted from the adsorbents and stored for subsequent analyses to determine if changes in chemical profiles could be correlated with insect age and levels of infestation. Changes in chemical profiles due specifically to infestation by Indianmeal moth larvae could lead to development of a highly sensitive detection method for infestations as a tool to minimize chemical use for insect control. Use of Vacuum to Disinfest Postharvest Tree Nuts of Insect Pests. Restrictions on the use of various fumigants have generated interest in non-chemical disinfestation treatments for postharvest tree nuts. Scientists at the USDA, ARS, San Joaquin Valley Agricultural Sciences Center, Parlier, CA, have developed non-chemical treatment protocols for tree nuts using vacuum applied to product within flexible containers. The most tolerant insect stage as well as the exposure times and temperatures necessary for adequate control were identified in laboratory studies. This information was applied in field trials which used the method to disinfest inshell almonds of navel orangeworm and Indianmeal moth. The work will provide growers and processors with a portable, low-cost, non-chemical method for disinfesting product, with treatment times comparable to those of phosphine fumigation. 4c.List any significant activities that support special target populations. None. 5.Describe the major accomplishments over the life of the project, including their predicted or actual impact. A parasitoid, Psytallia cf. concolor imported from the USDA-APHIS, PPQ, Moscamed laboratory in Guatemala City, Guatemala, was selected for potential biological control of olive fruit fly in California. Based on the success of this research, the California Olive Committee funded a regional parasitoid release project with collaborators from the USDA-ARS, Parlier, CA, and Gainesville, FL; APHIS-PPQ, Guatemala; and the California Department of Food and Agriculture, Sacramento, CA. A successful biological control program for olive fruit fly in California would help protect the $68 million canned olive industry in California. Control of overwintering populations of the navel orangeworm in pistachios is an important management technique, but current cultural methods are not completely effective. Field trials evaluated the efficacy of insect pathogenic nematodes against overwintering navel orangeworm, and established that the most effective nematode species was Steinernema carpocapsae at a practical concentration that growers could use. The development of effective application rates and guidelines for use of the nematodes should improve current field sanitation programs for navel orangeworm and eventually reduce pesticide applications against this pest. Mating disruption treatments were shown to significantly reduced crop damage due to navel orangeworm in almonds. Defining conditions under which mating disruption can be efficacious will hasten its adoption and reduce organophosphate insecticide and postharvest fumigation treatments. Temperatures often used to store tree nuts (32-41F) were found to effectively disinfest product of Indianmeal moth and navel orangeworm, the most important postharvest insect pests of dried fruit and nuts, in 2-3 weeks, but that the larval stages are very resistant to 50F. Dried fruit and nut processors will use this information to better control postharvest insect infestations and minimize the use of fumigants. 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? Technology was transferred to other researchers and industry counterparts via reprints, workshops, scientific meetings and other means of communication. Researchers cooperated with regulatory agencies such as APHIS, FAS, EPA, CalEPA, to resolve regulatory or quarantine issues. Technology pertaining to methyl bromide alternatives was transferred to shareholders, customers and cooperators via the annual Methyl Bromide Alternatives Outreach meeting, commodity group presentations, and user workshops. Specifically, research to develop control techniques for olive fruit fly in California was presented to growers, canners, and oil processors at meetings of the California Olive Committee, and to County Farm Advisors at the Olive Fly Management and Research Workshop. Almond, pistachio, and fig growers at their respective commodity group meetings were provided with USDA-ARS research findings concerning the development and use of a synthetic navel orangeworm pheromone, seasonal abundance of navel orangeworm; use of insect pathogenic nematodes to control overwintering navel orangeworm, efficacy of mating disruption for reduction of navel orangeworm damage, and potential of mass trapping for reduction of driedfruit beetle damage in figs. 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). Burks, C. S., Brandl, D. G., and Estrada, J. 2005. Control of driedfruit beetles by pheromone-enhanced mass trapping. Proceedings of the California Fig Research Institute; 2-17 Johnson, J. A. Vacuum treatments for postharvest dried fruits and nuts. Entomological Society of America, Pacific Branch Meeting. Monterey, CA. February, 2005. Johnson, J. A. Stored Product Insects: Identification and Biology. Pest Management in Food Processing 2005; seminar sponsored by Cardinal Professional Products Selma, CA June, 2005. Yokoyama, V. Y. and G. T. Miller. 2004. Systems Approaches for Control of Olive Fruit Fly: A Recent Introduction of an Exotic Pest in California. Abstracts of the XXII International Congress of Entomology, Brisbane, Queensland, Australia, August 15-21, 2004. Yokoyama, V. Y. and G. T. Miller. 2005. Systems approach for control of olive fruit fly in California. Abstracts of the FAO/IAEA International Conference on Area-Wide Control of Insect Pests: Integrating the Sterile Insect and Related Nuclear and Other Techniques 9-13 May 2005, Vienna, Austria, p. 249. Yokoyama, V. Y., Gina T. Miller, Gail E. Sergent, John Sivinski, and Pedro Rendon. 2005. Potential of Psytallia cf. concolor from Moscamed, Guatemala, for biological control of olive fruit fly in California. Abstracts of the 8th Exotic Fruit Fly Symposium 7-9 March 2005, Riverside, California, p. 25. Yokoyama, V. Y. Olive Fruit Fly Biology and Control Strategies. Olive Fly Management and Research Workshop, University of California, Davis, March 2005. Review Publications Siegel, J.P., Noble, P.M., Lacey, L.A., Bentley, W., Higbee, B.S. 2004. Use of nematodes for post harvest control of navel orangeworm (amyelois transitella) in fallen pistachios. California Pistachio Industry Annual Report, Crop Year 2003-2004. p. 113-114. Yokoyama, V.Y., Miller, G.T., Sivinski, J.M. 2002. Quarantine control strategies for olive fruit fly in california. Proceedings of the 6th International Fruit Fly Symposium, May 6-10, 2002, Stellenbosch, South Africa. p.241-244. Burks, C.S., Brandl, D.G. 2004. Seasonal abundance of navel orangeworm (Lepidoptera: Pyralidae) in figs and effect of peripheral aerosol dispensers on sexual communication. Journal of Insect Science [serial online]. 4(40). Available: http://www.insectscience.org/papers/4.40 Burks, C.S., Brandl, D.G. 2005. Quantitative assessment of insect pest damage to figs. Crop Management [serial online]. Available: http://www.plantmanagementnetwork.org/pub/cm/research/2005/figs/ Burks, C.S., Higbee, B., Daane, K., Bentley, W., Kuenen, L.P. 2004. Mating disruption for suppression of navel orangeworm damage in almonds. Proceedings of the 32nd Almond Industry Conference, December 1-2, 2004, Modesto, California. p. 1-10. Fields, P.G., Neven, L.G., Johnson, J.A. 2004. Practical alternatives to methyl bromide for use as quarantine and pre-shipment treatments in north america. International Conference on Methyl Bromide Alternatives and Emissions Reductions. September 27-30, 2004, Lisbon, Portugal. p 119-122, in (Eds) Batchelor, T. and Alfarroba. Johnson, J.A., Valero, K.A., Wang, S., Wang, J. 2004. Thermal death kinetics of red flour beetle, tribolium castaneum (coleoptera: tenebrionidae). Journal of Economic Entomology. 97(6):1868-1873. Tang, U., Wang, S., Mitcham, E.J., Johnson, J.A., Hansen, J.D., Hallman, G.J. 2004. Update on development of postharvest pest control treatments for nuts, citrus, and tropical fruits using radio frequency energy. Methyl Bromide Alternatives and Emissions Research Conference Proceedings. Oct. 31-Nov. 3, 2004, Orlando, Florida. p. 75.1-4. Wang, S., Johnson, J.A., Tang, J., Yin, X. 2005. Heating condition effects on thermal resistance of fifth-instar navel orangeworm (lepidoptera: pyralidae). Journal of Stored Products Research. 41(4):469-478. Johnson, J.A. 2004. Non-chemical alternatives for dried fruits and nuts: issues and opportunities. Methyl Bromide Alternatives and Emissions Research Conference Proceedings. Oct 31-Nov. 3, 2004. Orlando Florida. p. 74.1-4. Burks, C.S., Higbee, B., Kents, D., Bentley, W., Kuenen, L.P. 2005. Navel orangeworm abundance and damage in pistachios. California Pistachio Commission Production Research Report.143-153. Kuenen, L.P., Brandl, D.G., Rice, R.E. 2005. Modifications for trap assembly of pherocon 1c traps speed trap liner changes and reduce in-field preparation time. The Canadian Entomologist. 137(1):117-119.