Brief Summary of Minutes of Annual Meeting

Dr. Roberto Pereira called the meeting to order at 8:30 AM on February 12, 2006. He announced that the rewrite of the Regional Project went well and we have a new number S-1024. He also announced that the annual report is also complete. He also advised that the annual reports from all cooperators should be very brief in the future. He also pointed out that we need to select a Member-at-Large and the site for next years meeting. Dr. David Shapiro-Ilan, the local organizer gave details of the registration, facilities, services and meals to be served. Registration was set at $100. Dr. Roberto Pereira briefly discussed the Minutes of the 2005 meeting. SARES Administrative Advisor Dr. David Boethel gave report and praised participants for effectiveness in producing reports and proposal renewal. He also emphasized this group is very responsive to all his questions and does a lot of cooperative work. Dr. Boethel also talked about the Presidents budget and its implications. CSREES Administrative Advisor Dr. Robert Nowierski provided his report. He said that this group should put the meeting details on the NIMS system as soon as possible. He also suggested that this group should think about having the meeting in conjunction with another meeting. He then detailed new programs and funding sources including a new invasive species program. He then outlined several regulatory changes and discussed about the IPM training. After the brief introductions of the attendees the seminars began. The theme for this years meeting was the Use of Microbial Control in Major Acreage Crops. We had two seminar speakers. The first seminar was given by Dr. Don Steinkraus who presented Entomopathogens in Row Crops: A Candid Appraisal. The second seminar was given by Dr. Mike Adang, who spoke on Factors in the Insect Gut that Determine Susceptibility and Resistance to Bacillus thuringiensis. We then had discussion on the topic of the use of microbial control in large acreage crops. Research and extension needs were discussed. Enhanced involvement of government in facilitating registrations and simplifying regulations were suggested. Also expanded role of the government in mass-producing, and area-wide applications of microbial controls was discussed. The participants were asked to consider ways that microbials could be implemented in agricultural systems. It was suggested that a small-to-large approach was needed, where small crops are target first with later move to larger acreage crops. This is different from the current model adopted by many companies that seem to target the large acreage crops. It was explained that the reason for greater use of microbials in other countries such as Brazil is the direct involvement of the government and fewer regulations. The participants suggested the possibility of having microbial pesticides produced and distributed by cooperatives at a lower cost than that of commercial products. Government support may be necessary for funding and perhaps registration of such products The Small Business Innovation Research (SBIR) Program was discussed as a possible source of funding to develop and market microbial pesticides. Funds are available for different phases of product development. Other sources of funds for research and for implementation projects on microbial control of insects are commodity groups such as Cotton Incorporated, Soybean Promotion Board, and Sugarbeet Grower groups, which have funded projects of some of the participants. Groups interested in low chemical input alternatives for agriculture are also possible sources of funding and support for biopesticide projects. Other research ideas were discussed including questions to the speakers. Subproject discussions Discovery of entomopathogens and their integration and safety in pest management programs for major acreage crops. (Chair: Bob Behle). Informal oral reports were presented and the 25 attendees were encouraged to participate in discussions. In general, much of the information present focused on the use of fungal agents for control of pests of cotton, sugarbeets, sunflowers, and potatoes. Information was presented on control of insect pests on several crops using fungal pathogens. S. Jaronski reported on use of Metarhizium anisopliae for control of sugarbeet root maggot in laboratory and field trials, and added that this biocontrol agent was effective at low and moderate pest densities, but did not provide sufficient efficacy at high densities. Additional research with sugarbeets focused on integration of fungal control agents with cover crops and resistant plant varieties. For other crops, Jaronski reported that Metarhizium may be effective against red sunflower seed weevil adults but not against wireworms in wheat and potatoes. J. Leland reported on identification and production of new isolates of Beauveria bassiana at the ARS facility in Stoneville MS. Production of fungal conidial is sufficient to provide needs for larger field trials for control of Lygus bugs around field margins of cotton, to provide control before they infest the crop. M. Shapiro presented new information on additives consisting of natural products to improve the efficacy of baculovirus treatments. Improvements in efficacy by baculovirus could be useful for many crops beyond major acreage crops. Many side discussions erupted during this session and cooperative networks among scientists working on the various research topics were identified. Contributions by members who submitted written reports but were not in attendance were recognized. A summary of the reports and discussions would suggest that a majority of the efforts were focused on the use of broader host range fungal agents for control of specific pests in crops where conventional insecticide controls are not available or not effective. Discovery of entomopathogens and their integration and safety in pest management programs for medicinal, veterinary, and structural pests. (Co-Chairs: Jimmy Becnel and David Oi). Lee Solter (IL) discussed the occurrence of a microsporidian pathogen in a colony of lubber grasshoppers. Drion Boucias (Univ. of FL) reported on a non-occluded virus in housefly, which he is working on with C. Geden (ARS Gainesville). Virus is very contagious, host specific, and sterilizes female flies. Tim Kurtti (MN) discussed a Beauveria bassiana-like infection in black-legged ticks, Ixodes scapularis. Jimmy Becnel (ARS Gainesville) reported that transmission of a cypovirus (CPV) from the mosquito Culex restuans was enhanced by the divalent cation, magnesium, while calcium inhibited transmission. Mark Jackson (ARS Peoria) reported on collaborative research with ARS in New Orleans on termite control in trees. Roberto Pereira (ARS Gainesville) reported that a Thelohania isolated from a thief ant, Solenopsis carolinensis, was not the same species as T. solenopsae. He also discussed how transferrin may be involved in the immune response to pathogens in red imported fire ants. David Oi (ARS Gainesville) reported T. solenopsae is well established in four areawide fire ant IPM project sites in FL, OK, SC, and TX and the pathogen is spreading extensively in the TX site. A new nematode was found in fire ants by S. Porter (ARS Gainesville) and he is working with B. Hyman and E. Platzer (CA Riverside) on its identification. Jimmy Becnel and David Oi (ARS Gainesville) summarized submitted reports related to medical, veterinary, and structural pests from participants who did not attend meeting. Topics included nematode control of termites and bark scorpions (D. Gouge AZ; H. Kaya CA). T. solenopsae infection dynamics in different social forms of fire ants (J. Fuxa LA). Discovery of entomopathogens and their integration and safety in pest management programs for ornamental, vegetables, fruit and nut crops. (Chair: David Shapiro-Ilan). Larry Duncan is studying natural contribution of entomopathogenic nematodes in food webs. EPN application may increase populations of predatory fungi that can decrease populations of both EPN and plant parasitic nematodes. Metarhyzium may influence EPN predatory fungi thus can indirectly enhance EPN survival. Marty Shapiro is looking at plant products as potential UV protectants for microbial. They include lignins, medicinal herbs and teas. He advices people not to add calcium as a binder as it inhibits lignin binding. Patricia Stock has twenty new strains of nematodes from Arizona including 5 new species that are being described. Drion Boucias is working on Paeciolomyces for sucking insects in plant canopy. The fungus sporulates in the dark without a problem, and is good for white flies and scale insects. Beauveria genome is being sequenced. New Hirsutella sp. is a major pathogen of glassy wing sharpshooter. P. Grewal talked about long-term control of black vine weevil in ornamental nurseries using inoculative approach in grassy strips in between tree rows. He reported that a survey is being conducted in the muck soils area used for vegetable production. No EPNs have been detected in the production farms but EPNs are present in the grassy borders, shrublands, home lawns, and forests in the area. He also reported on the development of a novel delivery system for nematodes through the potting mix. Nematodes applied as infected cadavers of Galleria mellonella had better survival than the aqueous suspension. The desiccated cadavers were even better for nematode survival in the potting mix and garden soil. G. Jagdale talked about warm adapted nematode strains being tested for the fungus gnat control in greenhouses in Ohio. D. Shapiro-Ilan reported on further studies on EPN trait deterioration during culture indicating a genetic basis for deterioration and that the bacterial symbionts play a greater role in EPN trait change than expected. High levels of efficacy were reported versus peachtree borer in field studies using EPNs, and of pecan weevil using B. bassiana in trunk sprays (with novel UV protecting formulation from ARS, Peoria), and with M. anisopliae in trunk bands. Discovery of entomopathogens and their integration and safety in pest management programs for urban and natural landscapes. (Chair: P. Grewal). Bob Behle is developing and testing Oriental beetle lure (pheromone) plus fungus formulations. Lee Solter received APHIS Permit for the release of microsporidia for performing cage studies to determine the transmission of microsporidia. Predators of Adelgid beetles have two closely related microsporidia that could compete with each other. Bumble bees infected with microsporidia have been released in the United States. The bees could spread this to native bees. Patricia Stock is working on Steinernema diagnostics. Mitochondrial gene pcox1 and large subunit rDNA are better than ITS region sequences. Dawn Gouge is working on S. riobrave for the control of termites and scorpions. Don Steinkraus reported that Red Oak Forest is having oak decline disease. Insect larvae in the bark get into the hard wood. Jarred Leland produces Beauveria for tests against the Asian long horn beetle. Anna Legrand is working on the control of Black vine weevil and Japanese beetle with fungus. Stefan Jaronski is working on control of grasshoppers in rangeland management. P. Grewal reported on the funding of the Heterorhabditis bacteriophora genome sequencing project. He also discussed the progress on the metapopulation dynamics and gene flow in H. bacteriophora using pcox1 and ND4 genes and proposed a collaborative national study on this. Progress on white grubs control with nematodes was reported. Results of a survey for entomopathogenic nematodes on golf courses in Ohio were briefly presented. Final Business Meeting Roberto Pereira started the meeting by thanking the project participants for their contributions to the final report and the subproject chairs for preparing the different parts of the final report. The participants were asked to submit any last-minute changes to the draft reported which was circulated before the meeting. The chair also thanked David Shapiro-Ilan and Ian Brown for the successful organization of the meeting. In response to the chairs requests for comments on the meeting format several items were discussed including ways to enhance attendance at these meetings. It was suggested that we should budget some time for discussion in pathogen group as was done in the past, in order to attract those potential participants that may have been lost when recent changes in the project were implemented. The possibility of opening a bank account for the project was discussed. This would facilitate the deposit of carryover funds especially when they are needed to book the meeting site. Also, such account, in the name of a non-profit entity would facilitate the payment of registration fees by USDA and other federal employees, who face difficulties with the current system of making payments to a person. R. Pereira called for nominations or volunteers for Member-At Large. David Shapiro-Ilan and Patricia Stock agreed to serve if elected. The candidates were asked to leave the room while the other participants voted. David Shapiro-Ilan was elected member at large. Patricia Stock will take his place as co-chair for subproject 2. The members accepted the offer from Parwinder Grewal to hold next years meeting in Ohio. The theme for next meeting theme will be Implementing Microbials in Urban Landscapes and Forests. It will be held in Columbus, Ohio in conjunction with the National Turfgrass Entomologists Workshop in late February. With no other new business, the meeting was adjourned.

A catalogue of classical biological control introductions of entomopathogens against a great diversity of insects and mites has been prepared and submitted for publication. This was published in September 2005 as a Forest Service publication. During 2006 it should be put on the web. A paper summarizing the catalogue is being prepared. SUBPROJECT 1. Discovery of entomopathogens and their integration and safety in pest management programs for major acreage crops. Coordinator: Bob Behle, USDA-ARS-Peoria. Cooperating Agencies: AL, AZ, CA, MT, ND, NY, VT, USDA-ARS-Fargo, USDA-ARS-Peoria, USDA-ARS-Shafter, USDA-ARS-Sidney, and USDA-ARS-Stoneville. Reports by: Robert Behle, Sue Blodgett, Mark Boetel, Michael Brownbridge, Larry Campbell, J. E. Dara, Ann Hajek, Stefan Jaronski, Jarrod Leland, Michael McGuire, William Moar, Patricia Stock, and S. T. Ulloa. Significant research efforts continue to support development of microbes for pest control in large acreage crops and to verify the relative safety of these active agents. Research was conducted on corn, cotton, wheat, potatoes, sugarbeets, sunflowers and soybeans. The control studies have focused developing fungal agents, Beauveria bassiana and Metarhizium anisopliae, for control of Lygus in cotton and sugarbeet root maggot in sugarbeets, and identifying fungal pathogens infecting the invasive soybean aphid. Selectivity/safety of microbial agents was documented in cotton and corn by the absence of impact on non-target insect species. The development of resistance to Bt toxins remains a concern for transgenic crops, and research continues to identify specific mechanisms of resistance. Submitted reports identify cooperative research among six universities (Auburn University, Cornell University, Montana State University, North Dakota State University, University of California  Davis, and University of Vermont) and five ARS sites (Fargo, Peoria, Shafter, Sidney, Stoneville). SUBPROJECT 2. Discovery of entomopathogens and their integration and safety in pest management programs of ornamental, vegetable, fruit and nut crops. Co-Coordinators: David Shapiro-Ilan, USDA-ARS-Byron, and Robin Stuart, Univ. of Florida. Cooperating Agencies: FL, CA, USDA-ARS-Byron, USDA-ARS-Corvalis, USDA-ARS-Wapato. Reports by: Drion Boucias, Danny Bruck, Larry Duncan, Harry Kaya, Lerry Lacey, David Shapiro-Ilan, and Robin Stuart. Microbial efficacy was evaluated against various insect pests of fruit, vegetable, and nut crops. Experiments involved testing of novel pathogen species or strains, and new formulations or application technology, and ecology. Entomopathogenic fungi were the subject of several studies including a newly discovered pathogen Hirsutella homalodiscae nom. prov. against the glassy-winged sharpshooter, and Metarhizium anisopliae against Delia radicum. The potato tuber moth was found to be highly susceptible to volatiles produced by Metarhizium albus. A number of efficacy studies involved entomopathogenic nematodes including greenhouse and field trials with Heterorhabditis marelatus, Heterorhabditis bacteriophora, and Steinernema riobrave against the black vine weevil, the peachtree borer, the citrus root weevil, and the Andean potato weevil. Cocooned larvae of codling moth and Oriental fruit moth were controlled by Heterorhabditis spp. in fruit bins. Entomopathogen ecology and production technology were also focus of studies. The fungus H. homalodiscae was determined to be a major entomopathogen in the native range of glassy-winged sharpshooter, and in vitro production methods were developed for this fungus. The fungus M. anisopliae persisted well up to 133 days in five common soilless potting media. However composted manure decreased the prevalence of some nematophagous fungi and increased efficacy of entomopathogenic nematodes. Host exudates were found to affect entomopathogenic nematode infection dynamics. New technology was developed for production, formulation and application of potato tuber moth granulovirus, B. bassiana, and M. anisopliae. SUBPROJECT 3. Discovery of entomopathogens and their integration and safety in pest management programs for urban and natural landscapes. Co-Coordinators: Parwinder Grewal, Ohio State Univ., and Ed Lewis, Univ. of California-Davis Cooperating Agencies: AZ, CA, CT, GA, IA, IL, KY, NJ, NY, OH, OR, USFW-Corvallis, USDA-FS-Morgantown, USDA-FS-Hamden, USDA-ARS-Corvallis, USDA-ARS-Stoneville, USDA-ARS-Sidney. International Institutions: Bulg. Acad. Sci., Charles Univ. Prague, Forestry Inst. Slovakia, Fach. Eberswalde in Germany, BOKU in Austria. Reports by: Parwinder Grewal, Ann Hajek, Stefan Jaronski, Harry Kaya, Ana Legrand, Ed Lewis, Darryl Ramourtar, Lee Solter, Patricia Stock. Molecular data indicate that Nosema-type microsporidia in Malacosoma americanum from Illinois and Kentucky are probably the same species. Field studies of non-target effects of gypsy moth microsporidia showed that several non-target species become infected with Vairimorpha and Nosema. Transmission of a gypsy moth Nosema isolate in caged trees occurred even at low initial prevalences, and the transmission rate increased as initial prevalence increased. Horizontal transmission studies with N. lymantriae and V. disparis in gypsy moth showed that the first species to invade the host is preferentially transmitted to the offspring. The USDA-APHIS-PPQ and EPA approved the release of three species of gypsy moth microsporidia in Illinois. A microsporidium from the black vine weevil, Otiorynchus sulcatus, is being evaluated for its biological control potential due to its virulent and effects in late instar larvae, and the protozoan Crithidia sp. was recovered from several populations of bumble bees populations in Illinois. Several entomopathogenic nematodes (Steinernematidae and Heterorhabditidae) strains from tropical, desert and semi-desert habitats have been molecularly characterized and described. Ecological characterization of newly recovered entomopathogenic nematodes focused on several parameters including host range, foraging strategy, soil moisture and temperature and UV exposure. Entomopathogenic nematodes applied within cadavers were at least as effective or more effective, and survive longer, than those applied in aqueous solution. Heat- (35°C) or cold-shocks (1° and 10°C) increased trehalose accumulation and metabolism in H. bacteriophora, which may enhance nematode survival and provide desiccation protection. Isozyme patterns in 5 species of Heterorhabditi were species-specific and demonstrated a high degree of genetic variation among Heterorhabditis species, but relatively close genetic relationship between H. megidis and H. zealandica, and between H. bacteriophora and H. indica, and strong subspecies structuring in H. bacteriophora. Genetic variability of Asian and North American strains of Entomophaga maimaiga shows that North America strains are very homogenous and most closely related to strains from Japan. Asian and North American gypsy moths were equally susceptible to fungal strains from different geographic regions. Infected gypsy moth larvae mainly died in the afternoon and the fungus sporulated during the night. Metarhizium anisopliae (Strain F52 from Earth Biosciences) is being tested against Asian longhorned beetle in the USA and China, and against larvae of the Japanese beetle Popillia japonica in turfgrass. M. anisopliae Strain F52 and Beauveria bassiana Strain GHA used against mormon crickets caused up to 100% infection but field mortality was delayed due to low cricket body temperatures. M. anisopliae var. acridum isolates are up to 30 to 200-fold more infectious for Acrididae than the currently registered B. bassiana, and 3-7 times more infectious than B. bassiana GHA for the mormon cricket. SUBPROJECT 4: Development, evaluation and safety of entomopathogens for veterinary and structural arthropod pests Co-Coordinators: James J. Becnel, USDA-ARS-Gainesville, and David Oi, USDA-ARS-Gainesville. Cooperating Agencies: AZ, CA, CT, FL, LA, MN, MS, OK, SC, TX, USDA-ARS-Gainesville, and USDA-ARS-Peoria. Reports by: Theodore Andreadis, James Becnel, Drion Boucias, Chris Dunlap, Brian Federici, James Fuxa, Dawn Gouge, Mark Jackson, Harry Kaya, Timothy Kurtti, David Oi, Roberto Pereira, Edward Platzer, Sanford Porter, Steven Valles, and Charles Vossbrinck. Entomopathogens evaluated against veterinary pest included bacteria, viruses, fungi, microsporidia, nematodes and an insect pathogenic algae. The infectivity and pathogenicity of baculoviruses and cypoviruses were evaluated in laboratory bioassays demonstrating activity against a variety of important mosquito vectors and pests. A cytoplasmic polyhedrosis virus was isolated from the mosquito Culex restuans and its morphology, genomic, and molecular characteristics studies. Studies were conducted on a novel, non-occluded dsDNA virus from adult house flies (Musca domestica) that induces salivary gland hyperplasia and shuts down ovarian development rendering females unresponsive to mating. Various species of microsporidia have been sequenced to study relationships among the Microsporidia. The morphogenesis and replication dynamics of the different life stages of Helicosporidium sp., a non-photosynthetic, entomopathogenic alga were completed. A Beauveria species was isolated from black-legged ticks, Ixodes scapularis and sequence data indicates the isolate is closely related to, but to be distinct from, B. bassiana. Transposon based transformation vectors for genetic manipulation of endosymbiotic rickettsiae of ticks were developed. Studies found that various species of entomopathogenic nematodes were capable of infecting and killing the bark scorpion Centruroides exilicauda indicating the potential role of these parasitoids as a biocontrol option. For structural arthropod pests, several species of nematodes and a fungus were evaluated on various termite species. Nematode development was observed in several termites. The spread of the fire ant pathogen Thelohania solenopsae appears to predominate in the non-territorial social form of the red imported fire ant. A megaspore of T. solenopsae is implicated in transovarial transmission. Immune response of fire ants to pathogens may be associated with a transferrin gene. A new fungal pathogen was isolated from three ant species in Florida. Initial host specificity testing indicated that the fire ant pathogen Vairimorpha invictae was not infective to two cogeners of the red imported fire ant. Unidentified nematodes were found in worker caste ants of several colonies of Solenopsis fire ants collected in Argentina.

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