Brief Summary of Minutes of Annual Meeting

Meeting was called to order at 8:15 AM by Chairman Parwinder Grewal Neither the Administrative advisor (D. Boethel, SAES) nor the USDA-CSREES advisor were present or submitted reports for the meeting. At the 2007 meeting, a project was outlined for submission to funding agencies to study microbial control of grape pests. However, due to family issues, the project leader (Scott Costa) was unable to submit the final proposal to the granting agency before the annual deadline. Submission is expected to proceed this year. Participants were asked to consider serving as officers. A member at large and secretary are needed. Participants were also asked to consider potential locations and dates for the 2009 meeting. The low attendance at this meeting was partly attributed to the mid-week schedule rather than the historical Sunday-Monday schedule. The mid-week schedule makes it difficult for persons with teaching appointments to attend. Subproject 1. Discovery of entomopathogens and their integration and safety in pest management programs for major acreage crops. (Bob Behle, chair) Bob Behle (USDA-ARS-Peoria) reported that Lerry Lacy (USDA-ARS- Wapato, WA) is continuing to study a GV for control of the potato tuber worm. Mark Boetel (NDSU) reported on the application of Ma ((F-52) granule formulations applied for control of sugar beet root maggot. He also reported on cooperative work with S. Jaronski (USDA-ARS- Sidney, MT) evaluating combined treatments of Metarhizium anisopliae with seed treatments such as Poncho. There may also be an issue with Collembola reducing plant stands in sugar beet fields. Mark also reported looking at Metarhizium anisopliae in trap/cover crops for sugar beet root maggot control. Jarrod Leland (Novozymes) reported that his position in Stoneville MS was filled by Todd Ugine. Jarrod believed that Todd would continue treating wild host plants for Lygus control and also consider evaluating dispersion of Beauveria bassiana with Honeybees. The problem with Lygus is the insecticide resistance to many chemical pesticides. Jarrod also reported that applications of Metarhyzium anisoplae F-52 for control of wireworms provided a 20% increase in corn yield (J. Agron. 99: 1377-1381) Novozymes acquired a company with seed coating technology to improve seed germination. Lee Solter (Illinois Natural History Survey) reported that numbers of trained field crop entomologists is declining based on the reduced number of students in this area. Meeting was suspended for a short morning break at 10:00 am. The meeting was reconvened at 10:30 am with presentations by guest speakers Dr. Scott Pinkerton, USDA-ARS-NCAUR, Peoria, IL Titled: Potential use of baculoviral proteins in plant biotechnology to enhance insect control. The premise was that non lethal virus based proteins could be inserted into the plants genome to enhance the efficacy of virus based biopesticides. Robert S. Johnson, USDA-APHIS, Titled: Changes, challenges, requirements and procedures for importing and transporting microbial biological control agents. This presentation focused on the requirements for importation of foreign microbes and utilizing the relatively new electronic permit application system. The meeting was suspended for a lunch break at noon. Subproject 2 Discovery of entomopathogens and their integration and safety in pest management programs for ornamental, vegetable, fruit, and nut crops. (Robin Stuart & Patricia Stock, co-chairs; David Shapiro presiding due to absence of Robin & Patricia) Jarrod Leland (Novozymes) reported on Ma F-52 for controlling whitefly, mites, mealybugs, onion maggots, black vine weevil and onion thrips has shown benefits to yield. He expects a product launch in the Netherlands. Denny Bruck (USDA-ARS- Corvallis, OR) in cooperation with Novozome, demonstrated similar vine borer control using Ma and Bifenthrin in nursery container pots. Also, nematodes and Ma pot drenches worked well. He is also beginning to work on strawberries and will focus on soil insects, eg. Strawberry crown moth control with nematode applications. In cooperation with Joice Looper (plant pathologist), is looking at using a Pseudomonas species combined with an insecticidal active fit d gene to protect root rhizospheres. Parwinder Grewal (OSU) reported that two entomopathogenic nematode strains provide 60 to 80 % reduction in adult emergence of the grape root borer and that the nematodes remain near the vines for at least 2 years. For ornamental nursery pots, Steinernema carpocapsae and S. feltiae infect white grub larvae, but in the soil, temperatures can be too low for the nematodes while grubs continue to develop. He also reported that applications of entomopathogenic nematode infected G. mellonella near host plants, may induce a plant response that reduces infection by a plant pathogenic nematode. In vegetables, the goal is to follow entomopathogenic nematode population gene flow based on identified microsatellites. Mark Jackson (USDA-ARS-Peoria, IL) reported on cooperative work on Ma-52, whereby high carbon / low nitrogen fermentation produces microsclerocia, which can be dried, and applied. When they rehydrate, the microsclerocia form conidia that can infect pests. This concept is being tested against sugar beet root maggot. Lee Solter (Illinois Natural History Survey) is describing a microsporidia from the black vine weevil. She is also looking at bumblebee population fluctuations and reports that the Nosema in the US bumblebees is genetically identical to the European Nosema. Dawn Gundersen-Rindal (USDA-ARS- Beltsville, MD) is focusing on organic growers by identifying potentially functional organisms from resource isolates of bacteria and viruses. Jeff Aldrich (USDA-ARS- Beltsville, MD) works predominantly with pheromones and attractants with hemipteran insects and lacewings. He is also interested in the potential use of symbionts for insect control and cooperative work that may include pheromones with microbes for pest control. Robert Behle (USDA-ARS- Peoria, IL) reported the development of a lignin based adjuvant for tank mixing to provide protection from UV degradation. David Shapiro-Ilan (USDA-ARS- Byron, GA) reported on control of pecan weevil with trunk applications of B. bassiana and M. anisopliae fungal conidia. B. bassiana kills the host, but pecan weevils were still able to cause damage to nuts; M. anisopliae kills faster so may have an advantage. Very high levels of peachtree borer control were achieved with entomopathogenic nematodes. For the lesser peachtree borer, nematodes provide near 100% control when used with a moisture wrap. Control of plumb curculio is difficult and may require a trap crop for some commodities. Work continues on nematode production techniques in vivo through an SBIR grant with an industry partner, and cooperative work with Ed Lewis & Jim Campbell on nematode infection dynamics. Participants took a short break from 3:00 to 3:15 pm Subproject 3 Discovery of entomopathogens and their integration and safety in pest management programs for urban and natural habitats. (Parwinder Grewal, chair) Parwinder Grewal (OSU) reported that identified isolates of nematodes that are more pathogenic to white grubs have been licensed to Becker Underwood. Also, commercial lawn care companies typically do not use nematodes for grub control because the nematodes do not fit the control plan. Relative to Japanese beetle adult (lure and kill) control demonstrated that Steinernema carpocapsae is more effective than Heterorhabditis for adult control while the opposite is true for grub control. Following on research conducted by Shapiro-Ilan et al., inbred lines of nematodes were found to be more virulent to white grubs and inbreeding should fix genetics. Improved virulence was related to the ability of the nematode to escape after being encapsulated by the grub. Also, lipid metabolism is necessary for storage stability and some nematodes have been shown to have more genes related to lipid metabolism. Alfred Alumai (OSU) (working with Grewal) compared 5 lawn care programs from commercial to no input based on biological and aesthetic evaluations. Ordered ranking put Commercial = IPM > Organic > consumer > control; and costs ranking of Commercial > Organic > IPM > consumer > control. Ruisheng An (OSU) (working with Grewal) is studying nematode virulence and bacterial symbiont genes. Lee Solter (Illinois Natural History Survey) reported that she had not found suitable plots for the release of the microsporidia of gypsy moth. She is also working with the USDA Forest Service for control of the Hemlock wooly adelgid control. Jarrod Leland (Novozymes) is evaluating synthetic bands of M. anisopleae for control of the Asian longhorn beetle. In turf, the commercial M. anisopleae formulation is applied to control chinch bug (80 % control), billbug (60 % control) and white grub (20 % control). The oil formulation seems to get hung up in the thatch layer of the sod. Annual bluegrass weevil is difficult to control. Chris Dunlap (USDA-ARS- Peoria, IL) reported on the use of a foam formulation to apply Paecilomyces fumosoroseus blastospores for control of termites in trees. Cooperative research trials (with ARS-New Orleans, and the City of New Orleans) were established in New Orleans just prior to Hurricane Katrina, which flooded many of the treated trees such that follow-up evaluations are questionable. March 5, 2008 Subproject 4 Discovery of entomopathogens and their integration and safety in pest management programs for veterinary and structural arthropod pests. (co-chairs, David Oi and Jimmy Becnel; James Becnel, presiding) Mark Jackson (USDA-ARS- Peoria, IL) reported that future work using P. fumosoroseus blastopsores for termite control will focus on bait station applications because the balstospores are not repulsive to the termites. Lee Solter (Illinois Natural History Survey) reported that the medical entomologist (Novak) has left Illinois and that they hope to hire a replacement. The state continues to monitor for West Nile Virus from birds and Aedes japonicus has been found in Illinois. Parwinder Grewal (OSU0 has a student interested in studying WNV and the potential for mosquito production by waterscape environments. It was suggested that they contact Ted Andreadis. Jarrod Leland (Novozymes) reported the M. anisopleae product, Tick-X, has the data necessary for full registration, and has been shown to provide 50 to 90 % efficacy in trials in several northeast states. Trials were conducted in CT, NJ and NY. Registration for use in public lands (e.g., parks) would require more data. James Becnel (USDA-ARS- Gainesville, FL) reported on a new house fly virus, and that microsporidia infection interacted with adult mosquito behavior such that infected insects had lower levels of attraction to humans. Additionally, Jimmy Becnel reported on topical application of RNAi for insect control (patent pending). He also reported that two strains of Vairimorpha invictae have been identified from Florida and from South America. Jimmy Becnel reported that Boucias and Gedden are working on virus from tsetse fly that is also found in house flies. Sacs of rod-shaped virions are found in salivary glands. Feeding virus produced 40% infection in adults. Doesnt seem to infect maggots. They are doing fieldwork in stables. The virus is distinct from baculoviruses and distinct from the tsetse fly virus. It sterilizes adults- but how does this occur if only in salivary glands? Transmission isnt understood. It is not known if the virus is in any other tissues. Ellie Groden (Maine) has identified a new fungus for the European red fire ant. A mult-state pilot project for fire ant has been proposed. It has been demonstrated that colonies can be infected with Thelohania by transferring brood from infected colonies to new colonies. Phorid flies have been introduced for fire ant control. Lee Solter (Illinois Natural History Survey) reported that a microsporidia from the lubber grasshopper is related to a human pathogen and a field source was identified. It may be studied as a model system for drug screening against the human pathogen. Final Business Meeting Election of officers: Parwinder Grewal is the outgoing Chairman Edwin Lewis is the incoming Chairman to oversee the 2009 meeting. David Shapiro-Ilan is the Vice-Chairman. Denny Bruck was elected as member at large. Robert Behle was elected as Secretary. Mark Boetel agreed to serve as Subsection 1 co-chair 2009 Meeting Location: It was decided to ask Ed Lewis to host the meeting in Davis, CA. Ed Lewis subsequently agreed that he and Harry Kaya would organize the next meeting in Davis, CA on March 1 & 2, 2009. Theme should focus on Organic/Sustainable agriculture. Meeting time should revert to Sunday/Monday schedule. Meeting should be in February, but try to avoid Presidents Day Holiday. Funds: S-1024 should have a post-meeting balance of $300, currently held by Lee Solter 2007 Project As stated previously, the Crops At Risk (CAR) grant on Microbial IPM of Grapes was not submitted by Scott Costa in 2007, but is expected to be submitted in 2008. A question was raised as to whether the group could develop an information outlet for microbial control information. It could be in the form of a website based on fact sheets and could be managed through the microbial control division of SIP. The organization could be based on the current or future sections outlined by the S-1024 document. It was recognized that the site must not favor selected industrial partners or products. For the S-1024 project write-up, the advisors need to be contacted for report deadlines. Re-write ideas include IPM incorporating microbial control, and adding an extension/outreach component for the use of entomopathogens. It was agreed that the re-write will follow the suggestions of the SAES and CSREES advisors. The meeting was adjourned for lunch & a tour of the USDA-ARS-Peoria lab followed.

SUBPROJECT 1. Discovery of entomopathogens and their integration and safety in pest management programs for major acreage crops. [Chair: Bob Behle, e-mail: behlerw@ncaur.usda.gov] Research concerning microbial control of arthropod pests of major acreage crops is becoming limited as genetic engineering technology become widely accepted. Public and producer acceptance of Bt crops has allowed genetically engineered crops to become the focus of pest control for many of the major insect pests. Reports received for this subproject cover four crops (cotton, sugar beets, potatoes, and sunflowers) and identify the cooperation of two ARS locations (Yakama, WA; Sidney, MT; Peoria, IL) and two Universities (Auburn University; North Dakota State University). A population of cotton bollworm, Helicoverpa zea was selected for Bt Cry1Ac-resistance. Current resistance level is ca. 150-fold. There are numerous fitness costs associated with Bt resistance. Resistance appears to be co-dominant. Resistant insects still cannot fully develop on Bt cotton. There is still no evidence for field-evolved resistance in Bt cotton for tobacco budworm and cotton bollworm. (Auburn University) A new formulation of Metarhizium granules, based on hitherto undescribed microsclerotia produced under certain liquid culture conditions, was developed by ARS for use against soil dwelling insects. These granules show superior sporulation and efficacy against sugarbeet root maggot in laboratory, soil-based bioassays. A patent application was submitted. Field trials against low-moderate maggot pressure showed root damage reduction equal to the chemical standard, terbufos. Against intense attack, however, the fungus provided no root protection, verifying past experience. (ARS  Sidney MT, ARS  Peoria IL and NDSU  Fargo ND). In a second year of field testing, Beauveria bassiana Strain GHA was not efficacious against adult red sunflower seed weevil. Regardless of whether delivery of conidia to the flowers was in aqueous suspension or as a dust, rates of 1x109 conidia per flower caused only minor infection and mortality of the targeted weevils. (ARS-Sidney MT and NDSU). The potato tuber moth (PTM) granulovirus (PoGV) and Bacillus thuringiensis Berliner subsp. kurstaki (Btk) were evaluated for season-long control of PTM on potato foliage in 2007. Compared with untreated controls, 10 weekly applications of PoGV (1013 OB/ha) reduced PTM populations by 86-96% on foliage and 90-97% on tubers added to cages shortly before harvest. Infection rates of 82 - 95% were noted within individual larval cohorts. Equivalently timed Btk treatments (1.12 kg product/ha) were significantly less effective at population suppression, with a 36-76% reduction in larvae recovered from tubers added to cages. A PoGV/Btk alternation was significantly more effective than Btk alone and as effective as PoGV. Bioassays were conducted to evaluate the residual activities of foliar deposits. Early season applications were highly effective for the first 24 hours (e 93 % mortality) with a steady decline in activity over 10 days. Studies on the effects of PoGV and Bt on PTM in stored tubers is continuing. (ARS-Yakama, WA) The effects of volatiles produced by the fungus Muscodor albus on potato tuber moth larvae within infested tubers inside sealed chambers were also studied. The length of exposure to M. albus significantly affected mortality of larvae. Exposure durations of 3, 7, or 14 days at 24°C followed by incubation in fresh air at 27°C until emergence resulted in mortalities of 84.2, 95.5 and 99.6%, respectively. Effects of M. albus on larval survival was also monitored at 10, 15 and 24°C, using an exposure duration of 7 days followed by incubation in clean air until emergence. Mortality of larvae was sharply reduced at the lower temperatures resulting in 50.8, 76.8, and 95.4% mortality, respectively. Longer exposures resulted in production of unacceptable levels of CO2 for tuber storage. (ARS-Yakama, WA) SUBPROJECT 2. Discovery of entomopathogens and their integration and safety in pest management programs for ornamental, vegetable, fruit, and nut crops. (Robin Stuart & Patricia Stock, co-chairs) Insect pathogens are used extensively in these particular crops, often due to their high market values. A number of successful efficacy trials indicate substantial potential for microbial control agents in peach and pecan orchards. In peach orchards, during 2007, tree-limb damage from lesser peachtree borer (Synanthedon exitiosa) was reduced by 80-100% following applications of Steinernema carpocapsae; these high levels of efficacy were only achieved when the nematodes were applied using a protective gel or bandage [USDA-ARS Byron, GA; University of Florida; University of GA]. In a pecan orchard, up to 90% suppression of adult pecan weevil (Curculio caryae) was observed following trunk applications with B. bassiana or Metarhizium anisopliae [USDA-ARS Byron, GA; University of GA; USDA-ARS Peoria, IL; Novozymes, Inc.]. In laboratory research, metabolites from Xenorhabdus and Photorhabdus bacteria were found to suppress key plant diseases of pecan and peach [USDA-ARS Byron, GA]. Additionally, a new nematode species, Heterorhabditis georgiana was isolated in Georgia and found to have some promise in biocontrol potential [University of Florida; USDA-ARS Byron, GA; Fort Valley State University, GA]. Several enteric bacteria isolated from the Colorado potato beetle inhibited the growth of the entomopathogens Photorhabdus temperata and Beauveria bassiana in vitro. (MD) The effect of simple (crop) and more complex (crop plus refuge) habitats were compared on the persistence and movement of the entomopathogenic nematode, Steinernema carpocapsae, applied to soil in a maize field. During the 30-day experiment, S. carpocapsae was detected up to 3m from the application site. The dispersal pattern in both habitats is best described with an exponential probability distribution, with nematode detection decreasing with distance from the application point. In the first year of the experiment, greater movement occurred within complex habitats than in simple habitats. In the second year of the experiment, S. carpocapsae movement was similar in both habitats. This result may be due to the comparable plant density and soil moisture in both habitats, in contrast to differences in these variables in the first year. The results indicate that S. carpocapsae movement is not necessarily dependent on plant diversity, but may respond to variation in plant density, and subsequently, soil moisture. Complex habitats may serve as movement corridors for S. carpocapsae, depending on the contrast between the refuge and crop area. (PA) Populations of weeds, soil quality characteristics, and economic performance in organically-managed corn in minimum and conventional tillage systems were measured. We detected three genera of entomopathogenic fungi in the experimental plots: Metarhizium anisopliae, Beauveria bassiana, and 2 species of Isaria. Over the 3-year transition period, detection of M. anisopliae declined. There was a non-significant trend for detection of M. anisopliae to be greater in full-till (moldboard plow) systems compared with minimum till (chisel plow) systems. We hypothesize that greater spread and mixing of the soil in full till treatments resulted in greater detection of M. anisopliae compared with minimum till treatments. Other biological and physical differences in soil conditions in the two tillage treatments may have also resulted in differential survival of the fungus or its hosts. (PA) The dead adult psyllids killed by H. citriformis (ARSEF 8315) were found attached to the underside of citrus foliage in Polk, Hendry and Marion counties, and those killed by P. fumosoroseus (ARSEF8316) were collected in Polk county. Hundreds of psyllids were collected that were killed by H. citriformis, but only a few psyllids killed by P. fumosoroseus were found. This difference in abundance is probably due to the fact that psyllids killed by H. citriformis are tightly attached to the citrus foliage by the synnemata, but those psyllids killed by P. fumosoroseus are only loosely attached and may be removed from the tree by wind and rain. Two pilot field trials using this strain of P. fumosoroseus as a microbial insecticide resulted in reduced populations of psyllid nymphs. However, economic and technical issues, coupled with the frequent use of copper applications to control citrus canker and other plant pathogens, likely will prevent the commercial development of this beneficial fungus for use in Floridas citrus pest management programs. We do not yet understand the role of these two fungi on Asian citrus psyllids in Floridas citrus groves. Research is needed to determine how the pathogens are transmitted, when the beneficial fungi are most abundant, and how widespread these psyllid diseases are distributed in Florida. Quantitative surveys also are needed to evaluate the effect of each pathogen on psyllid populations throughout the year. So far, we have identified two new natural enemies in Florida that can kill Asian citrus psyllids and gained knowledge about the basic biology of each pathogen in our laboratory studies. (FL) The development of an accurate bioassay of efficacy of EPNs in the different citrus soils will allow citrus growers to determine whether they can use these biological control agents against the larval stages of Diaprepes. As we have demonstrated that the commercially available, S. riobrave, can move through soil effectively in many different soils to a depth of 20 inches, this information is useful if the larval stages occur from the soil surface to 20 inches below. We found that sandy soils are best for EPNs and soils with high clay contents are more problematic. The nematodes should be effective against larval stages that occur in potted material. However, trials are needed under California conditions to demonstrate that nematodes are efficacious against Diaprepes. (CA) Work continues on using EPNs against the navel orangeworm in pistachios. Efficacy has been good, and many growers are adopting the methods developed by Siegel for applying EPNs via irrigation systems during cooler winter months. We are currently following the long term fate and impact of EPN applications. In particular, we are documenting changes in nematode and microarthropod fauna after applications. (CA) SUBPROJECT 3. Discovery of entomopathogens and their integration and safety in pest management programs for urban and natural habitats. (Parwinder Grewal, chair) During 2007, we conducted our third year of studies on the dispersal of fungal and viral pathogens of gypsy moth along the lead edge of gypsy moth spread, in central Wisconsin. We are presently analyzing data from the 3 years of the study to estimate the minimal gypsy moth density associated with presence of Entomophage maimaiga and Lymantria dispar nucleopolyhedrovirus (LdMNPV). We have now introduced E. mamaiga to the two of the Apostle Islands in Lake Superior, northern Wisconsin to conduct experiments to test our dispersal estimates from central Wisconsin; based on 2 years of data, weve shown that E. maimaiga does not already occur in these areas. LdMNPV is still abundant in gypsy moth populations on Basswood Island, one year after Gypchek was sprayed for control. (NY) We conducted our first year of studies of the metapopulation dynamics of E. maimaiga, using study sites in central Pennsylvania. We sampled populations on three different spatial scales throughout the last field season, repeatedly caged larvae in the field and we have begun experiments to compare activity and persistence of E. maimaiga resting spores in different types of soils. Studies of the ability of three bird species (that eat gypsy moth larvae) to disperse LdMNPV were conducted using field cages. Feeding behavior of the birds had a strong impact on local spread of the virus. (NY) In addition, data on bioassays of different species of anamorphic hypocrealean fungi against Anoplophora glabripennis (ALB) adults and larvae were analyzed and published. Data on the reduction in fitness of A. glabripennis females before death due to Metarhizium anisopliae were also analyzed and published. Studies of the LC50 of M. anisopliae F 52 against A. glabripennis adults were completed and results will be published in 2008, in association with data on the persistence of activity of fungal bands in the field in New York State. Finally, we completed and published results from studies examining naturally occurring environmental contamination with M. anisopliae from fungal bands in the field but also due to A. glabripennis adults after they have walked across fungal bands. SUBPROJECT 4: Development, evaluation and safety of entomopathogens for veterinary and structural arthropod pests Coordinators: James J. Becnel, David Oi, ARS-Gainesville Entomopathogens evaluated against veterinary pest included viruses and microsporidia. Genomic analysis of MdSGHV, a non-occluded, enveloped, rod-shaped, double-stranded DNA virus responsible for salivary gland hypertrophy (SGH) in adult houseflies, revealed a genome of ~120 kbp with 119 potential ORFs. Comparisons of the available identifiable ORFs suggest that this virus is distinct from baculoviruses. Cytological and ultrastructural characteristics were examined for a Vavraia culicis-like microsporidian species isolated from Aedes albopictus in Florida and subspecies status is proposed for this isolate. Studies with Aedes aegypti infected with Edhazardia aedis found a reduction in mean human host attraction and landing/probing rates compared with rates in microsporidia-free females. E. aedis-infected mosquitoes required 20% more time than healthy females to bite skin treated with 15% DEET. For structural arthropod pests, necrophoresis by the invasive European fire ant, Myrmica rubra, may play a role in protecting colonies from infection. New fungal pathogens were identified from M. rubra. A new virus was found and characterized from the red imported fire ant, Solenopsis invicta. Microsporidian fire ant pathogen V. invictae was not detected in fire ant parasitic flies developing in V. invictae-infected S. invicta, further confirming host specificity. Nematodes repelled termites from termite bait stations perhaps resulting in less termite mortality. Various species of microsporidia have been sequenced to study relationships among the Microsporidia including Ovavesicula popilliae, a microsporidian parasite of the Japanese beetle, Popilliae japonica and a novel microsporidium from the saw toothed grain beetle Oryzaephilus surinamensis.. Veterinary Pests MdSGHV, described initially as a non-occluded, enveloped, rod-shaped, double-stranded DNA virus, is responsible for the salivary gland hypertrophy (SGH) symptoms observed in feral populations of adult house flies. Infection by this virus sterilizes female house flies, since adults displaying symptoms of SGH show no sign of ovarian development. Recently, utilizing Nycodenz gradients, MdSGHV was purified from hypertrophic glands of infected house flies. Negative staining revealed enveloped virus particles measuring 65 by 575 nm that possessed unique spiral grooved surface topology. Analysis of its genomic DNA using end-labeled restriction fragment length polymorphism revealed a ~120 kbp genome. Sequencing data from two titration lanes of the 454 pyrosequencing produced 1.4 million bases (>10-fold coverage) that were consolidated into 17 contigs ranging from 512 to 40,170 bp. Sequence information obtained from the ends of the MdSGHV EcoRI library was used to further connect them into three contigs that through PCR amplifications resulted in a single circular 124,254 bp genome. The genome contains 119 methionine initiated potential ORFs with 50 or more amino acids and minimal overlapping (SoftBerry- fgenesV). BLAST analysis identified ~29 ORFs (e-values <0.1). Analysis of the chemical data from LC MS/MS from the bands extracted from SDS-PAGE gel of enveloped virus revealed homology to 32 ORFs from the SoftBerry data base The vast majority of these structural ORFs represent novel proteins, with only four having homology to sequences in GenBank. Comparisons of the available identifiable ORFs suggest that this virus is distinct from the baculovirus genera. (FL) Comparative small subunit rDNA sequence analysis was completed on Ovavesicula popilliae, a microsporidian parasite of the Japanese beetle, Popilliae japonica. This parasite represents a distant sister group to Paranosema and Antonospora, parasites of grasshoppers, and collectively form a second major group (the Nosema/Vairimorpha clade representing the first) of microsporidia which infect terrestrial insects, suggesting independent origins for both groups. Phylogenetic analyses of Ovavesicula and other microsporidia having a multi-sporous sporogony reveal that this condition is found in several unrelated taxa implying either that multi-sporous sporogony is the ancestral condition for microsporidia or that it has multiple origins (CT). A detailed description was made on the cytological and ultrastructural characteristics of a Vavraia culicis-like microsporidian species isolated from Aedes albopictus (Scuse) in Florida . This Florida isolate, is the only known isolate of a species of the genus Vavraia from mosquitoes propagated in laboratory culture. Although the Florida isolate has been used under the name Vavraia culicis in several molecular phylogeny and host-parasite studies, it has not been structurally characterized and its relationship to the type species Vavraia culicis has never been examined. Structural data strongly support placement of the Florida isolate within the genus Vavraia and indicate its close relationship to both the type species of the genus and to other Vavraia-like mosquito microsporidia to which the name V. culicis has been applied. However, the identity of the Florida isolate with V. culicis (Weiser, 1947) Weiser, 1977 cannot be presently confirmed. Morphometric examination of spores of several Vavraia-like microsporidia isolates from mosquitoes, including the type material of Vavraia culicis, indicates that Vavraia culicis- like microsporidia probably represent not a single species, but a group of closely related organisms. Subspecies status is proposed for the Florida isolate. (ARS-Gainesville) We investigated whether sick/diseased mosquitoes manifest differences in their host seeking and blood-feeding related behaviors compared with healthy mosquitoes, and the extent to which such differences impact the effectiveness of topical repellents such as DEET. Infection of Aedes aegypti with Edhazardia aedis reduced mean human host attraction and landing/probing rates in female mosquitoes by 53 and 62%, respectively, compared with rates in microsporidia-free females. Infection with E. aedis reduced the average weight of unfed female mosquitoes by 4%, caused them to imbibe 23% less blood, and to lay 30% fewer eggs than healthy females. In contrast, E. aedis-infected mosquitoes required 20% more time than healthy females to bite skin treated with 15% DEET. Statistically significant morbidity in E. aedis-infected females was indicated by reductions in host attraction and landing/probing responses, the mass of unfed and blood engorged females, and fecundity, and by increased DEET repellency. (ARS-Gainesville) Structural Pests Studies exploring the role of necrophoresis in defense of ant colonies against infection were continued, demonstrating that the invasive European fire ant, Myrmica rubra, preferentially selected areas with full exposure to solar radiation for depositing the cadavers removed from their nests and infected cadavers exposed to ambient solar radiation for 48 hours did experience reduced sporulation when transferred to high humidity conditions favorable for spore production. A new fungus isolated from M. rubra in England has been confirmed as a pathogen via reinfection assays and is currently being described as Paraisaria myrmicarum sp. nov. with collaborators in England (H. Evans) and USDA-ARS (J. F. Bischoff). An additional two fungal isolates have been recovered from the same site in England and represent new host records and the first isolation of these species outside of the tropics. They have been considered distinct varieties: Hirsutella stilbelliformis var. myrmicarum var. nov. and H. subramanianii var. myrmicarum var nov. (ME) The multi-state (FL, MS, OK, SC, and TX) IPM project for fire ant suppression in pastures involving the microsporidian pathogen Thelohania solenopsae, concluded in 2007. T. solenopsae was well established in all sites with red imported fire ants, Solenopsis invicta, but did not establish in the MS site which contained black imported fire ants, Solenopsis richteri. Project was expanded to include higher value demonstration sites such as quail habitat, schools, and recreational parks which have a greater potential to adopt the utilization of combining biocontrols and insecticidal ant baits. (ARS-Gainesville). The fire ant pathogen Vairimorpha invictae was not detected by PCR from fire ant parasitic phorid flies (Pseudacteon litoralis) that developed in V. invictaeinfected fire ants. This is further evidence that V. invictae has a narrow host range. (ARS-Gainesville). A new virus (tentatively named Solenopsis invicta virus 2 [SINV-2]) was discovered in the red imported fire ant. SINV-2 has a monopartite, single-stranded RNA genome that contains multiple open reading frames in the sense orientation. The genome has been sequenced in entirety and is comprised of 11,303 nucleotides with a polyadenylated region at the 3' terminus (GenBank Accession Number: EF428566). Phylogenetic analysis of the genome and the proteins it encodes revealed unique placement of SINV-2 exclusive from Dicistroviridae, iflaviruses, Picornaviridae, and plant small RNA viruses. Impact of the virus on fire ant populations is being investigated. (ARS-Gainesville). Preliminary field tests applying Steinernema carpocapsae to subterranean termite bait stations, has indicated significant short term avoidance of stations by Heterotermes aureus and Gnathamitermes perplexus. In laboratory bioassays, repellency studies have indicated higher dose rates may lead to lower mortality rates due to increased repellency effects. (AZ) Small subunit rDNA sequencing and host range studies have been completed with a novel microsporidian parasite from the saw toothed grain beetle Oryzaephilus surinamensis. The parasite appears to be a species of Paranosema based on phylogenetic analysis and is designated as P. oryzaephili. Host range studies demonstrate that this microsporidium is more infectious to the rusty grain beetle Cryptolestes ferrugineus that it is to the type host (CT, ARS-Manhattan, KS).

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