Brief Summary of Minutes of Annual Meeting

Business Meeting Opening Remarks - S. Arthurs (2014 chair): Welcome to all and begin with introductions (23 participants). Minutes for the 2013 meeting had been reviewed and posted on the NIMSS Website. Minutes for this meeting are required to be posted within 60 days for the report. Chairs Report - S. Arthurs: Participants were happy with the new format of the working group meeting coinciding with the ESA national meeting. We had a second member symposium accepted for this 2014 ESA meeting entitled “Thinking outside the box: Entomopathogens in IPM systems”. The purpose of the working group was provided for several new members. ESA provided registration waivers for several student speakers. AES Administrative Advisors Report - B. Rogers Leonard: Background to project was provided. Project purpose is to provide opportunities to establish multi-state collaborations for research. In some states, funding for travel for members of land grant universities. “S” designates this as a Project initiated in the Southern region. This is an open meeting, but requires an active project and an annual report for each project year. He emphasized the impact of multistate projects. To become a member, delegates can register through the NIMMS site with approval from their state experiment station. This project ends September 2017. It was recommended that this group consider submitting a nomination for the USDA-NIFA’s Southern Region Project Award. In previous years Southern project groups working on pecan and sweet potato won this award. Four years ago the Southern project on soybeans won the national award. There is a history of success. National Program Leader- Robert (Bob) Nowierski: Bob provided information on a USDA NIFA grants program called the ‘Crop Protection and Management Program’ (CPPM) that could provide funding opportunities for the S1052 group. This grant program is comprised of three major elements: the Applied Research and Development Program – ARDP (R.. Nowierski directs), the Extension Implementation Program (Marty Draper directs) and the Regional Coordination Program (Herb Bolton directs). The ARDP program will fund three types of projects: Applied Research only; Research-directed projects (must contain at least 20% extension); and Extension-directed projects (must contain at least 20% research). The Request for Applications (RFA) for the program is expected to be posted on the NIFA website (www.nifa.usda.gov) in January 2015. Applications may only be submitted by colleges and universities. However, ARS scientists can obtain funding through a sub-contractual arrangement. Contact Bob Nowierski at: Rnowierski@nifa.usda.gov if you have any questions about this grant program. Educational Opportunity - Lee Solter: Reported on an educational short course in Insect Pathology directed through the University of Illinois. Currently 1-week and 1-semester graduate courses are provided. Lee Solter, Ann Hajek, Lerry Lacey and Rich Humber will offer a beginner course in Insect Pathology at Cornell University in the 2nd week of June 2015. They are trying to obtain IOBC support. If there is interest, an 8-week course can be offered in the region from University of Illinois at about $500 per student. It is unclear whether students from other institutions could get credit for this. Bob Nowierski suggested Lee take examine the ‘IPM cubed’ online courses (Integrated Pest Management Education at the University of Minnesota), which are approved for certified crop advisor CEUs (found at: www. umn.edu\IPM3). Lee and research associate Wei-Fone Huang are currently undertaking a microsporidia sequencing project (using the SSU rRNA gene) in order to archive the Illinois Natural History Survey collection in GenBank., If anyone would like live spores from the collection, please contact Lee. The next 1052 working group meetings were scheduled for 2015 (Minneapolis, MN) and 2016 (Orlando, FL). The group settled on the symposium themes ‘Manipulation of Host Behavior to Enhance Microbial Control’ (2015) and ‘Regional Status of Microbial Control Programs’ (2016). Elections will be undertaken at the 2015 meeting.

Accomplishments Subproject 1. Annual Crops Jaronski: Wheat stem sawfly larvae were collected in several states and 30 fungal isolates additional to the original ones, were obtained. Looking at endophytic Beauveria bassiana, Jaronski has indirect evidence that GHA strain can become endophytic in wheat, coincident with reports from Texas A&M that GHA can be deliberately made endophytic in cotton. Met 52G microsclerotia have been licensed by Koppert from USDA and a European patent on the technology granted (U.S. patent still pending). Field evaluations of fungi against wireworms had some encouraging results in 2014; both Beauveria GHA and Metarhizium brunneum F52 on corn grit granules were applied in furrow; wireworms may be attracted to sporulated granules applied at seeding. Band over row sprays at planting were also effective in protecting yield. Both these projects need post docs, so Stefan made an announcement to anyone who knows of interested candidates. Funding is provided by Montana State University and the Wheat Commission. Dara: Has been working to integrate IPM with clientele farmers as a Strawberry and Vegetable Crops Advisor in California. Currently most growers only rotate chemicals with different modes of action but do not employ biological pesticides and biological control agents. He has been testing Beauveria bassiana GHA (Botaniguard®) and Met 52 (he only works with registered microbials) in field trials against mites and other pests. Results so far look good showing the potential of microbials in the strawberry IPM program. B. bassiana was also shown to increase strawberry yield and growth during parts of the production season. Dara has also seen positive results using microbials against green peach aphid; plants look healthier and stronger. Trials in raised beds also showed that strawberry plants were healthier compared to the control plants where B. bassiana was applied as root dip and soil treatment. Last year he applied Beauveria GHA, Isaria and Met 52 to cabbage and found Beauveria to be best possibly due to the microbials aiding to combat water stress. He stressed the importance of demonstrating the success of these microbials in field trials, since farmers need proof. Work on stink bugs: organic growers are having a hard time controlling this pest and the hunt for natural enemies is a key priority. Dara will visit India to look for natural enemies. Leland: Reported that costs and application rates are critical to the successful commercialization of microbials. Has been working with plant growth-promoting rhizobacteria (PGPR). His company is currently screening several thousand microbes for impacts of plant growth and resistance to pests and diseases. Three major Novozymes company goals are 1) to explore avenues for all existing Met 52 products; 2) A new company approach to the discovery of novel strains/isolates is to grow up about 2000 microbes and test these in the field. If any positive results are seen by a particular strain then it will be developed further; 3) They are also working on foliar sprays. Jurat-Fuentes: Has been evaluating corn and soybean pest susceptibility to Bt through Cry toxin interaction with insect midgut receptors. Currently working to develop new toxins as alternatives to current Bt toxins that are at risk of becoming ineffective due to resistance. Fall armyworm with field-evolved resistance to Bt corn were demonstrated not to display cross resistance to commercial Bt sprays. A Bt-enhancer, an insect receptor (cadherin)-based synergist of Bt toxins, provides a way to overcome Bt resistance and increase killing of relatively Bt-tolerant insects. Work has been done with this Tenebrio peptide that enhances activity against coleopteran pests. Kudzu bugs (Megacopta cribaria) are also a big pest problem in the South on soybeans. Currently working on developing gene silencing through RNA interference as a potential to control this pest. A colony of kudzu bug has also been started if anyone is interested in obtaining these insects. Chase Metzger wondered how effective these Bt sprays might be against blackheaded fireworm (Tortricidae), which have been a serious pest on cranberries. The pest infests and pupates in the tip of the plant and in really bad infestations can eat the entire plant except the stem. He has tried Entrust®, Pyganic®, Grandevo®, Venerate®, Dipel® and Met 52 granules in his cranberry plot trials. It was discussed that Met 52 granules might not persist well in the boggy cranberry fields, as was found by a Finnish researcher (VÄNNINEN, I., TYNI-JUSLIN, J. & HOKKANEN, H. 2000. Persistence of augmented Metarhizium anisopliae and Beauveria bassiana in Finnish agricultural soils. BioControl 45: 201-222). Mascarin: Has been working with Mark Jackson (USDA), finishing his PhD on a fermentation method to increase blastospore production and stability of B. bassiana which allows for much faster and less extensive production compared with traditional solid substrate fermentation of conidial formulations. Only some strains of Beauveria can produce blastospores, others only produce hyphal strands. Spray-dried product has good shelf life, which is 6 months at 28°C. Provisional patent is pending. Microsclerotia of Metarhizium have been tested to have a one year shelf life when vacuum packed. It may be possible to produce Metarhizium blastospores (strains from Brazil) but more work is needed. Kobori: In Mark Jacksons lab, has been working to develop microsclerotia for Trichoderma fungus. Trichoderma microsclerotia have now been produced using liquid fermentation. Shelf life more than one year was confirmed. Provisional patent is pending. Boucias: Has been working with Burkholderia bacteria that are associated with the caecum and ovaries of chinch bugs. These bacteria are thought to act as detoxifying compounds for their host. Bacteria can be acquitted horizontally or vertically (ovary). A ten-fold difference in the quantities of gut bacteria between individuals have been observed, which may be correlated with insecticide resistance, which would represent a new mechanism of insecticide resistance. Drion traveled to Japan recently to speak with Kakuchi et al. regarding Burkholderia. Avery: Has been working with yellow margined leaf beetle (chrysomelid) – an important pest of crucifers in the southeast region. Reported that the fungus Isaria fumosorosea is not effective against this pest, possibly due to the production of glycosinolates from cole crop hosts (antagonism). Some efficacy against eggs and early instars was observed in the laboratory trials. Work is ongoing investigating the effect of Isaria on reducing the fecundity of the female after becoming infected. Molting has been observed to be hindered after infection with Isaria. Beauveria bassiana isolates from Brazil can kill this insect species. Spined soldier bug is used as a predator in this system and is obtained from Canada, it was tested. Cultural control remains an important IPM tool in this system, if the 1st and 2nd leaves are broken off the plant, this can significantly reduce egg infestations by yellow margin leaf beetles. Organic farmers are looking for help! Subproject 2. Perennial Crops Koppenhöfer: Previously, we surveyed for native entomopathogenic nematodes (EPNs) in NJ highbush blueberry fields and surrounding native pine areas that also contain blueberries and related species. Steinernema glaseri was the only species isolated from cultivated blueberry. In the native areas two other species were found rarely, but the dominant species was a potentially new species close to S. glaseri. DNA studies suggested enough difference from S. glaseri that we will conduct further work to determine its species status and characterize its potential as a control agent. Jaronski: Has also been looking at controlling spotted wing Drosophila in tunnel crops: blackberries and raspberries using various combinations of Beauveria GHA + spinosad sprays, F52 microsclerotia and Met 52 EC. This is cooperation with Tanigoshi and Gerdeman at WSU, Mt. Vernon WA. The late instar larvae drop to the ground to pupate in the soil and the idea is to create an infectious mine field to ultimately reduce the adult population in the next cycle. The pupae can be infected but it’s difficult to tell because they don’t display signs of sickness for up to 5 weeks sometimes. Goble: Has been working with F52 microsclerotia to control Asian longhorned beetles. Incorporating these microsclerotia granules into hydromulch [a mixture of water + wheat straw mulch + psyllium tackifier], represents a novel mycoinsecticide that can be sprayed onto the trunks of forest or orchard trees to control insect pests. The fungus applied in hydromulch using psyllium (as a tackifier 6-8%) in paint sprayer equipment, can significantly reduce beetle fecundity and survival when they are exposed to sprayed logs and bark surfaces, with profuse spore production. Logs that received Mb MS-hydromulch produced more conidia and the median survival times of beetles were lower (18-22 d) in these treatments compared with logs that received no hydromulch (median survival 23-26 d) and had less conidia, which increased the survival time. Hydromulch can persist for up to 100 days in the field, with sometimes very little material loss due to weathering. Spore production continues to increase over a 30 day period depending on increased moisture (rainfall, RH) and temperature, as well as time. Overall beetle mortality when exposed to wood pieces in the various sprayed replicates ranged from 36-93% depending on the production of conidia within the hydromulch, which was ultimately dependent upon the weather. Mafra-Neto: A chemical ecologist researcher, CEO and Director of Research and Development at ISCA Technologies, Inc. which he founded in 1996 in Riverside, California, a company specializing in the development of semiochemical solutions for pest management, robotic smart traps and nanosensors. Insect pathogens may be able to be incorporated in his companies patented wax pheromone release technology (SPLAT) especially if combined with insect attractants. He joined the meeting to collaborate and help other members with formulation needs in their existing projects. Verle Rodrigues: Reported on work in Puerto Rico screening endemic B. bassiana strains against coffee berry borer (CBB). Coffee is grown in the mountains (sensitive habitats) in Puerto Rico, which represents a huge challenge for control options hence the need for microbials. Some farmers report around 28% crop loss due to the pest, while other famers report between 60-80%. Currently only Beauveria GHA commercial strain is registered for control against this new invasive pest. Monitoring and surveys are currently undertaken on 200 farms. NEPAL: Rodrigues worked in Nepal recently with coffee white stem borer (cerambycid, Xylotrechus quadripes) that is an important pest in the Southeast Asia region. The larvae spend all of their time inside the coffee stem and are difficult to control. In Nepal, up to 80% crop damage is reported so there is reason to be alarmed about this pest. Finally it was reported that a number of mites are also important pests: Brevipalpus spp, which can transmit a virus and red palm mite, which is a pest of coconut product. They are looking at spraying these pests with entomopathogenic fungi but targeted applications are difficult. Shapiro-Ilan: In pecan an Organic Transitions project is targeting pecan weevil with nematodes, fungi and Grandevo®. Beauveria bassiana applied with composted manure in a pecan systems appears to enhance fungal persistence. The soil around the pupal cell of pecan weevil was found to be inhibitive to B. bassiana (potentially protecting the insect from infection, mechanism yet unknown). In peaches (working with Bob Behle and others), high levels of efficacy against peachtree borer and lesser peachtree borer are obtained using Steinernema carpocapsae; the sprayable gel Barricade® can be used to protect nematodes from environmental extremes aboveground or in soil (e.g., in lieu of irrigation). Targeting plum curculio, an integrated sentinel tree approach using Steinernema riobrave as a soil treatment appears promising. Metabolites from nematode bacterial symbionts are showing promise for control of various plant diseases in peach and pecan. Working with Ed Lewis and others, entomopathogenic nematodes were found to exhibit aggregative dispersal behavior (thus moving through soil perhaps akin to a pack of wolves). Also, it was reported that a new book has been published entitled: “Mass production of beneficial organisms: invertebrates and entomopathogens” (Elsevier) Edited by Juan Morales-Ramos; M. Guadalupe Rojas; David Shapiro-Ilan. Boetel: - Reported on the latest in furrow applications with Metarhizium anisopliae (MA1200) and Metarhizium brunneum (F52) spores formulated on corn grit (collaboration with Jaronski) for control of sugarbeet root maggot. These fungal granular formulations are thought to be more appropriate for growers controlling sugar beet root maggot than conventional organophosphates. Growers are currently still mainly using chlorpyrifos, terbufos and thimet which are very toxic to the environment. Tests conducted with fungal granular formulations and a new resistant variety of sugar beet in 2014 were encouraging becasue the fungal treatments did not differ significantly from the chemical standard. This is the first successful sugarbeet field trial in a number of years. Subproject 3. Natural and Urban Landscapes Koppenhöfer: We showed that EPNs can provide control (average 65%) of moderately high larval populations of the annual bluegrass weevil (ABW) on golf course fairways. However, control rates were about 20% lower than for standard insecticides and tended to further fall behind at high ABW densities. In 2013 we observed that nematodes- imidacloprid combinations gave additive ABW control. To examine the effect of split application on efficacy of nematodes and nematode-imidacloprid combination we conducted a field study in 2014. The nematode Steinernema carpocapsae and imidacloprid were applied alone or in combination or half rates either as single or split applications. All treatments except imidacloprid alone provided significant control. Nematode imidacloprid combinations but not nematodes alone were significantly better than imidacloprid alone. Split application of nematodes whether for nematodes alone or combinations showed greatest potential. Where imidacloprid is already used for white grub control, its combination with split nematode application could be an effective option for ABW larval control. In spring applications, a product based on the entomopathogenic fungus Beauveria bassiana was ineffective against ABW adults and larvae but synergized strongly with the pyrethroid insecticide bifenthrin against pyrethroid resistant ABW. B. bassiana did not interact with the neonicotinoid imidacloprid against ABW larvae. B. bassiana was ineffective against young oriental beetle larvae. It did however synergize with a product based on azadirachtin, but control levels were moderate. Jaronski: Grasshoppers in rangelands- 150 B. bassiana and 150 Metarhizium isolates (from D. Roberts) have been screened during the past 5 years in a program to identify candidates for grasshopper control. None have shown good, economical field efficacy.. Field release of M. acridium (commercialized in Africa and Australia), which is an effective pathogen and specific to Orthoptera, is still not allowed in Unites States because the species is not indigenous. However, data collected during past two years indicate this fungus cannot overwinter in the Northern Plains. Efforts are still underway to secure APHIS permits for field testing. , Grasshopper pathogens collected locally have not had any major effect in controlling these pests because the insects exhibit a behavioral fever. Still recovering new B. bassiana isolates from grasshoppers for population genetics analysis DNA analysis (so far 731 isolates have been collected from 5 locations in 2014, with prevalence of infections among living grasshoppers ranging from 14-56%). In contrast only one Metarhizium isolate was isolate from grasshoppers in 2014. New Chinese Bt for locust control is available and patented in China only (a company in China has the license to mass-produce this locust-active strain of Bt). ARS Beltsville and Sidney MT are currently screening the vast ARS Bt collection for locust-active isolates. Lima Caetano: To control Sirex noctilio, the nematode Deladenus siricidicola Kamona (imported from Australia), which sterilizes female wasps, has been extensively and successfully used as a biological control agent in the Southern Hemisphere. Curiously, a non-sterilizing (NS) strain of D. siricidicola is commonly found in North America and it is thought that the NS strain was introduced with S. noctilio when this woodwasp was introduced to North America. Finding an appropriate biological control agent in North America has been challenging due to the existence of native species of Sirex woodwasps that are not considered pests but are part of the decomposer community in forests. Therefore, evaluation of biological control agents requires studies of host specificity of the nematodes. At Hajek lab, we evaluated the NS strain of D. siricidicola, which is poorly understood and is a potential competitor of D. siricidicola Kamona. D. siricidicola has two forms: a form that parasitizes S. noctilio and a mycophagous form that feeds on the Sirex fungal symbiont Amylostereum. The goal of this study was to compare associations between the NS and Kamona nematodes and different isolates of the symbiotic fungus, mainly to evaluate the ability of the nematodes to develop and reproduce when feeding on different isolates of Amylostereum associated with Sirex in North America. We found that both strains of nematodes grow better when feeding on the three strains of A. areolatum that are assumed to have been introduced to North America with S. noctilio and they did not grow on A. chailletii. Comparing the growth of the two nematodes on different fungi, we saw significant difference on A. areolatum BD, which favors D. siricidicola NS growth. O’Donnell: Representing work done at the University of Maine-a post doc is working on the recovery of USA collected Hirsutella isolates from European fire ants. Other work by Eleanor Groden has looked at the bacterial symbionts of a mermithid nematode found in ants. Kaitlyn is looking at the development and survival of winter moth on lowbush blueberry in relation to other host plants (cherry, oak, apple, maple and birch). NPV viruses have been found in winter moth caterpillars. Verle Rodrigues: Spoke about the bacterial symbiont in false spider mites, which are thought to be gut symbionts. They are using next generation sequencing to determine frequency and abundance of Cardinium, as well other symbiont associated to plant mites. He also mentioned Wolbachia, Burkholderia bacteria found in coffee berry borer in Puerto Rico, which feed on coffee. This is amazing since caffeine is toxic to insects. Leland: There was a brief report back on the successful use of the granular product Met 52 used against vine weevil in Europe and chinch bugs in Canada. Avery: It was reported that Talus® (an insect growth regulator) + IFR (Isaria fumosorosea fungus) were shown to be compatible for combined use and control of Madeira mealy bug. After fungal infection, the nymphs can shed the Isaria spores (IFR) through the molting process until the adult stage. Once adults are infected, they will succumb to the infection quickly and die, also eggs in the ovisac are susceptible to these treatments. Steven Valles, Sanford Porter, and David Oi (ARS-Gainesville): Fire ant pathogens Kneallhazia solenopsae and Solenopsis invicta virus-3 as well as fire ant decapitating phorid flies, were released in the Palm Desert area of California with the assistance of the Coachella Valley Mosquito and Vector Control District. Releases were made in May and June 2014 in urban, irrigated habitats. All three agents have been detected in late October, thus surviving the summer desert climate. More releases and monitoring to confirm establishment and spread are planned for 2015. In a cooperative project with the Foundation for the Study of Invasive Species (formerly USDA-ARS South American Biological Control Laboratory) over 180 colony fragments of S. invicta, and colonies infected with the microsporidium Vairimorpha invictae, were collected in Argentina and hand carried to the ARS quarantine facility in Gainesville, FL. Colony fragments are being screened for new pathogens in an effort to discover new bio control agents, and V. invictae infected colonies are being used to develop methods to produce inocula. This is the first shipment from Argentina since 2012, because of difficulties in obtaining Argentine export permits. Arthurs: Reported that a couple of his graduate students were presenting at the ESA meeting. One was working on screening commercial nematodes against grass webworm (Crambidae) in turf. Another student was working on testing pathogen mixtures against thrips in roses.