Brief Summary of Minutes of Annual Meeting

General Comments Opening remarks and Chair Report (Denny Bruck): Welcome to all and we will begin with introductions (21 participants). Minutes for the 2012 meeting had been reviewed and posted on the NIMMS Website. Minutes for this meeting should be posted within 60 days as part of the final report. The Committee improved awareness of the working group from the Entomological Society of America (ESA) as indicated by our proposed symposium being selected as a Section Symposia. Advisors Report (Rogers Leonard). There are 24 members listed on the NIMMS website as associated with this working group. The purpose is to provide funding for travel for members of land grant universities and is designated as part of the southern region. This is an open meeting, but requires an active project and an annual report for each year of the active project. Robert (Bob) Nowierski is the Federal NPL for this group and is making a presentation at the ESA Symposium. This project ends September 2017. The group might consider support of community gardens and projects supporting water quality, both federally supported missions. The group was able to garner funding to support travel of invited speakers for the symposium, $1250 for Nicolai Meyling. The meeting report is due 60 days after the meeting. The report may include relevant publications as additional support of the interactions and accomplishments of this working group. Additionally, grant titles, and authors could be listed. Also the biocontrol short course will be listed. Group History (Denny Bruck): Meeting dates were recently (since 2012) changed to coincide with the ESA meeting to garner additional support and recognition while reducing travel costs for members who can attend both meetings simultaneously. Additionally the change supports the ability of the group to submit and support symposium titles for the ESA meeting. We have a need to update the member and email lists. Lee S. will attempt to coordinate the S1052 working group email list with the Society for Invertebrate Pathology. Connections will help to foster networking among members. Committee Elections: Each member serves a 2 year term before proceeding to the next position. Denny Bruck steps down after 2 years as chairman and passes this leadership position to Steve Arthurs (former vice chair). Surendra Dara transitions to Vice-Chair from Member-at-Large (MAL). Robert Behle was nominated and elected as the new MAL by vocal vote. Tarryn Goble and Pasco Avery were nominated to fill the Secretary position. Tarryn was narrowly elected as Secretary. 2014 Meeting: We plan to hold the next meeting in Portland, OR, on 15th November 2014, the day before the ESA meeting. We decided to sponsor another member symposium at the ESA Conference, with the theme: ‘Thinking outside the Box: Incorporating Microbials into IPM Programs’. New business: No new business was introduced. The meeting was adjourned at 6 p.m.

The subproject reports summarize discussions of ongoing and new research efforts. This was the first report for the new project. Progress was made for all three objectives. Subproject 1. Annual Crops (field crops, vegetables, etc) Co-Chairs: Mark Boetel and Bob Behle Mark reported recent research using Metarhizium microsclerotia granules for control of sugar beet root maggot (in collaboration with Jaronski). Late planting for the 2013 growing season put application timing out of synchronization with the insect’s life cycle. Metarhizium anisopliae strain MA1200 isolated from sugar beet cyst nematode (Heterodera schachtii) performed better than commercial F-52 strain (opposite of lab bioassay results) possibly because of better survival in the soil (Stefan Jaronski). Alternative to at-planting applications are band-over-row sprays of the fungus can be timed with the insect’s immigration into sugar beet fields to oviposit. The sprays are intended to place spores in the immediate area of egg laying and neonate migration to the roots. However significant water (80 gallons per acre) is required to sufficiently wet the soil, even with a 5 inch band. Efforts are currently underway to “piggyback” the Metarhizium application with herbicide or disease control applications, as well as, soil-incorporated insecticides, primarily the chloronicotinyls. Insecticidal activity in laboratory bioassays is currently estimated at 250,000 conidia per CC of soil for an LC50. The maximum application rate of any kind of granular formulations of fungus is considered to be 20 pounds per acre. As for research on the benefits of cover crops, the best results have been achieved with cereal grain cover crops intended for wind erosion control, which also helped chemical pesticide control and encouraged fungal infections. For wheat, control of wireworm is a concern in cereal crops. Beauveria is being evaluated for control of wheat stem sawfly (with collaborator Gadi Reddy, Montana State University). Fourteen isolates of Beauveria, isolated from post diapause larvae still in their hibernacula, may be endophytes in wheat and provide a form of host plant resistance for control. They are currently conducting a fall survey to determine the spread of Beauveria endophytes infecting sawfly across the Montana wheat region, to be followed by a more extensive spring survey. In addition, experimental work has started examining potential of establishing Beauveria endophytism in a spectrum of wheat varieties, using several wheat stem sawfly-derived isolates. Ed Lewis reported that Symphylans are damaging organic strawberries and vegetables in California. A request was made for assistance to develop suitable control measures. John Umble (OSU) was suggested as a contact for assistance. One suggestion was to use tobacco as a green manure treatment. As part of a project with the correctional system, Surendra Dara studied the impact of Beauveria bassiana on insect pests of strawberries. No pest infestations were detected to measure the impact, but there was significant growth promoted by Beauveria bassiana. Yield data was not collected. Root dips at planting were expected to promote plant growth and establish B. bassiana in the environment. Beauveria is being incorporated into IPM control of insects, reducing chemical applications and to manage development of resistance by Lygus bugs. Some combined treatments are similar to positive responses from chemical control applications. Progress in control of spider mites was obtained when applying reduced rates of chemicals with Beauveria. Fungicides are often applied, and the application timing and compatibility among treatments to strawberries needs further evaluation. Also, thrips control measures in lettuce are being evaluated, as well as spider mite control with Beauveria ES (1 qt/A) with Acramite. Marrone Bio Innovations has a new product, Venerate (MBI 206), that controls spider mites. Bagrada Bug is a new/emerging pest in cole crops and preliminary assays show good potential of B. bassiana, Isaria fumosorosea, and to some extent M. brunneum. Parwinder Grewal reported that the Organic transition project is evaluating crop rotations, and tillage practices relative to commercial organic farming. Cover crops include radish, rye, and clover. Galleria spp. were used to bait entomopathogens and evaluate nematode diversity as a bio-indicator of ecosystem maturity among treatments. First year results show no EPNs, Beauveria, was common. Juan Jurat-Fuentes found that soluble cadherin protein mixed with Bt products enhanced activity of Bt. Efficacy improved from 10 to 90% mortality. Potentially, this may be used to expand host range of Bt products or lower production costs. Enhancement may be in the form of improved “speed” of activity. The effect was observed over different insect Orders. Industry has not yet shown interest in the technology. An additional contact may be Valent Biosciences. A colony of armyworm with field-evolved resistance to Bt corn has been established and displayed cross resistance to other cry proteins in transgenic crops, but not to commercial Bt pesticides. Robert Behle reported that Blapstinus spp. beetles are a pest of melons in southern California. Damage to the crop is in the form of feeding on the rind. Efforts to control adult beetles with granules containing Metarhizium microsclerotia have been largely unsuccessful in the field, even though laboratory assays demonstrate adults and larvae are susceptible to infection by the fungus. Hot dry environmental factors may inhibit effective control. Stefan Jaronski commented that in Ecuador, weevils migrating to potato plants are being controlled by Beauveria-contaminated traps that provide shade and catch migrating beetles. This is an example of successfully using entomopathogenic fungi and the insect behavior to develop a control strategy. Combined biological methods are being evaluated for control of cabbage root maggot in a collaborative project with the Swedish Agricultural University (Nicolai Meyling). Combined applications of parasitoids (Trybiographa rapae) and fungus (Metarhizium brunneum) provided an additive effect in caged field experiments. Canada has certified use of honeybees to carry Beauveria to flowers as an application method (Stefan Jaronski). Similar concepts exploiting insect behavior include use of male-attractive traps to inoculate males which then transfer inoculum to females during mating at a later time. Subproject 2: Perennial Crops (orchards, small fruits, forage, etc.) Co-Chair: Ed Lewis Ed Lewis reported that persistence of EPNs in pistachio as relates to soil type is being studied in an effort to develop methods for improved navel orange worm control. Larvae overwinter in nuts on the ground and are controlled by nematodes (mainly S. carpocapsae) applied through the irrigation system. Persistence of EPNs has been shown to last about 10 weeks after applications in February. Better persistence was observed on the north side of tree. S. carpocapsae are eaten by a specific mite, but also infect earwigs. Particle size and salinity are important soil factors for nematode persistence. Grape root borer was controlled by the application of nematodes using the “cadaver” formulation (Parwinder Grewal). Specialized application equipment (disk cut trench) has been developed and persistence is currently being evaluated. Storage of the cadaver formulation may be an issue due to contamination by saprophytic fungi. Metarhizium and Beauveria are being evaluated for Drosophila suzukii fly control on cane berries, particularly those grown in tunnels, targeting the larvae as they drop to the ground to pupate (Stefan Jaronski). It was questioned if nematodes would infect pre-pupal larvae. Steve Arthurs stated that citrus greening disease remains a problem in Florida. Greening will ultimately be controlled by resistant rootstock. However, Isaria fumosorosea blastospores may be used for control of the psyllid vector, achieving about 50% population reduction in our tests. Our experiments with Pasco Avery show that I. fumosorosea blastospores interact with various adjuvants used in citrus production. I. fumosorosea was persistent in the field for 21 days on infected psyllids. Psyllids stop feeding within 2 days when infected (Pasco Avery). Related data from L. Lacey demonstrated lower plant disease (zebra chip) transmission (to potato) from psyllids that were treated with entomopathogenic fungi including I. fumosorosea. A paper in has been recently published on compatibility of oils with I. fumosorosea. David Shapiro Ilan‘s research demonstrated longer persistence of fungal agents applied as a part of compost for organic or conventional production of orchard crops (cooperator = Wayne Gardner). Peach tree borer can be infected by nematodes with high efficacy levels. Lesser peach tree borer attacks trees above ground and the use of Barricade can be used to maintain moisture necessary for nematodes to seek the larvae of LPTB (cooperators from the group include Bob Behle & Ed Lewis). High concentrations of Barricade require dual application; lower concentrations are being studied for a single application with Barricade and nematodes combined (and showing promising results thus far). Ed Lewis’s student indicates that Barricade at 0.5% v/v allow the nematodes to orient to the host, whereas a thicker Barricade concentration inhibited orientation of the nematodes. Plum curculio suppression is being explored through an integrated system using trap trees to attract insects and then controlling them in the soil with nematodes. The current expectation is that approximately three trap trees per hectare will be sufficient. Several field trials in the Mid-Atlantic States and New England indicate that plum curculio suppression was achieved using nematodes without irrigation. Entomopathogenic nematodes responded directionally to electrical current, magnetic fields and electric field. It has been demonstrated that the nematodes migrate in mass groups. Red bay ambrosia beetles are attacking avocado trees in Florida and vectoring the laurel wilt disease (Pasco Avery). Collaborators (Daniel Carrillo et al.) have evaluated commercial fungal pathogens for control of this beetle. Field collected beetles may have endemic Beauveria at a relatively high proportion in the population. Subsection 3 – Natural and Urban Landscapes (turf and ornamental, urban, forests) Co-Chairs: David Oi and Steve Arthurs David Oi reported that fire ant virus has been harvested from ant colonies and can be applied using various baits. The virus is found naturally in the US. Efforts continue toward importing a microsporidian pathogen of fire ants, Vairimorpha invictae, from Argentina as a classical biocontrol agent. Both organisms cause lateral infection of colony members. The tawny crazy ant, an invasive ant from South America, also has a potential viral pathogen. The webworm (Lepidoptera: Crambida) is an important lawn pest in the southeast and is susceptible to nematodes (Steve Arthurs). A commercial product Millennium® based on S. carpocapsae is being evaluated by a graduate student. An invasive thrips species (chilli thrips) is a major pest of roses in the southeast and may develop insecticidal resistance. The pest attacks new leaves and buds. A project with the LSU AgCenter (Yan Chen) evaluated biorational insecticides in combination with fertilizer treatments to manage the pests in nurseries. I. fumosorosea product Pfr97 20% WDG has shown potential for controlling an invasive whitefly pest in Florida (Pasco Avery). Lab experiments demonstrate fungal infection of nymphs and adults. Sirex (wood wasp) nematodes have provided control of this pest in South Africa (Ann Hajek). It has a symbiotic fungi for larval food. Sirex invaded, bringing a nematode pathogen, which is non-sterilizing nematode and provides poor control. Classical control concerns were raise about the impact of this non-sterilizing nematode on native sterilizing Sirex species. As a result, there will be a focus on studying the biology of this system. Tarryn Goble indicated that Asian longhorned beetle eradication is not complete. USDA-APHIS is supporting imidacloprid for control of the beetle, but titer is too low for adequate control. Since fungal banding are too labor intensive, hydromulch with fungal agents (M. brunneum microsclerotia) is being evaluated for sprayable application to trees. Rehydration may cause additional conidia production. Adding 3% psyllium (Plantago) (as a tackifier) helps Hydromulch applications stick to the trees but decreases conidia production. Female beetles die more quickly than males. Leellen Solter reported that microsporidia are a problem in coccinellid production and research on this problem was being funded by a USDA Forest Service grant. Exotic beetles are released in large numbers for control of woolly adelgid infesting eastern Hemlocks. Unfortunately, microsporidia have been released with the beetles, though the pathogen remains at low field prevalence. Immune responses of gypsy moth being challenged with naturally occurring pathogens are being studied. The Gypsy moth genome is being evaluated with proteomic data to determine gene response. As for research on bees, Koppert is working to verify absence of Nosema pathogens in colonies mass-reared for pollination services. Honeybee microsporidia were determined to only attach the gut and current control with fumagillin has resulted in enhancing microsporidia production in the laboratory. In Europe, invasive Harmonia axyridis ladybeetles are considered a nuisance pest in parks (Nicolai Meyling). A graduate student is looking at fungal infections of H. axyridis and have found mostly I. farinose in summer. These counts were correlated with high counts on leaves of Tilia sp. trees which are the habitat of H. axyridis in the study area. Shaohui Wu is studying control of masked chafer grubs with EPN and reported specific-variable control under laboratory conditions (Hb > Hm > Sf > Sr). Interactions of EPN with fungi (Ma and Bb) and showed an additive effect for insect control. Field trials were inconsistent. Granule Ma dissolved in water was better than the EC formulation. Interactions were complex: decreased Ma germination and some nematodes may have shed their sheath to escape fungal infections. In England and the Netherlands, pine processionary moths cause an allergic reaction due to urticating hairs (Dave Chandler). Sprays of Bt have been applied in field areas. Metarhizium and Beauveria need to be tested. Also, research is being conducted on control of thrips in strawberries to develop basic information about biocontrol applications. Stefan Jaronski reported on grasshopper control in rangelands. APHIS has raised questions about persistence of exotic Green Muscle (Metarhizium acridum) fungus and its spread under field environments. Long-term persistence tests under permit from APHIS indicate that infectious residues of the fungus persist only about 2 months in the summer and do not persist through winter months into the next year, on the Northern Plains. Currently, 400 isolates of Beauveria have been collected from living grasshoppers in MT and ND. Based on the Bloc gene sequence they represent a wide variety of clades. About 150 have been screened for transient high heat tolerance (6 hours at 37C or 41 C) of vegetative mycelium, simulating one day of grasshopper behavioral fever, as well as characterized for growth rates over the range of 9-34 C. About 1/3 of 150 Metarhizium spp. isolates, chosen from among several thousand from grasshopper breeding habitat soils across western U.S. made by Don Roberts, Utah State University, immediately resume growing after 6 hours at 37 degrees C, but only M. acridum resumes immediate growth after 6 hours at 41 degrees C (equivalent to a day of behavioral fever). A key to M. acridum success seems to be its ability to resume growth immediately when temperatures decrease to 20-28 degrees C such as in the evening. The other fungi have a latent (no- growth) period after exposure to the two high temperatures, from 12 to 96 hours. Research is also being conducted to discover a Bt that is active against locust (Cry 7 Ac), based on reports from China. The extensive ARS culture collections are being screened using molecular probes for the appropriate gene and subsequent bioassays of potential candidates (collaboration with ARS, Beltsville MD). The goal is to apply the Bt

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