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S301: Development, Evaluation and Safety of Entomopathogens for Control of Arthropod Pests

Annual/Termination Reports (SAES-422): [04/16/2001] [04/29/2002] [04/01/2003] [04/13/2004] [04/21/2005]

Date of Annual Report: 04/16/2001

Report Information:
  • Annual Meeting Dates: 02/25/01 to 02/26/01
  • Period the Report Covers: 01/2000 to 12/2000

    • Participants:
    Brief Summary of Minutes of Annual Meeting:
    Action Items/Assigned Responsibilities/Target Dates:
    1. The meeting was convened at 9:00 AM by Chairman James J. Becnel.

    2. Harry Kaya reported on local arrangements.

    3. Minutes from the 2000 annual meeting were accepted.

    4. Chairman, Becnel reported: The major activity by the Chair, Officers and Project Chairs was the preparation and submission of a new project to replace multi-state project S-265. This involved compiling a critical review of S-265 Regional Project ?Development and Integration of Entomopathogens into Pest Management Systems?, 1995-1999 and preparation of a new project ?Development, Evaluation and Safety of Entomopathogens for Control of Arthropod Pests?. The new project was approved on September 28, 2000 by Dr. George Cooper, CSREES, and established as MRF Project 301. This was only possible through the considerable efforts of the writing committee, which consisted of Dr. Lee Solter (AES, Illinois), Dr. Stephen Wraight (ARS, NY), Dr. Rosalind James (ARS, TX), Dr. Parwinder Grewal (AES, OH), Dr. Mike Kline (ARS, OH), Dr. David Oi (ARS, FL) and Ted Andreadis (AES, CT). Dr. David Boethel (Administrative Advisor, LA) was also crucial in seeking reviews and expeditiously processing the proposal through the various administrative channels to gain approval prior to expiration of the current project. Thanks goes also to the membership for responding, on short notice, with the necessary information when requested in order to meet our deadlines.


      Dr. Harry Kaya (AES, CA) is also to be commended for his excellent organization and preparations in hosting the 2001 Annual Meeting of S-265 in Davis, CA.

    5. Dr. David Boethel (SARES Administrative Advisor) presented his report to the members and introduced the web-based reporting for the project.

    6. Dr. Rick Meyers (CSREES Administrative Advisor) presented his report to the members on the role of CSREES, new hires and positions within CREES, and an overview of the budget.

    7. A site selection committee was formed with Member at Large, Roberto Pereira, chair, and John Vandenberg


    Summary of the Discussion:

    1. Talks were presented by Dr. Drion Boucias, University of Florida, titled "Microbial Mining of Insect Pathogens", and Dr. Desmond R. Jimenez, AgraQuest Inc., titled "Discovery of Microbial Metabolites at AgraQuest Inc."

    2. Subproject 1: Development, evaluation and safety of entomopathogens for leaf feeding insect defoliators (Co-Chairs: Stephen Wraignt and Leellen Solter). Several ongoing and potential research collaborations were discussed including projects on 1) genetic diversity of pathogen species and regional populations (one focus being on Beauveria bassiana), 2) pathogen-pathogen interactions, 3) tritrophic interactions (including effects of Bt transgenic plants on natural enemies), 4) laboratory and field development of microbial pesticides for specific pests such as diamondback moth, Colorado potato beetle, and Japanese beetles, 5) the role of microbial control agents in pesticide resistance management, 6) formulation and use of nematodes for foliar pests, 7) work on control of pest complexes in specific cropping systems such as vegetables and cotton, and 8) application technology. It was suggested that investigators with ideas for research collaborations bring proposal outlines to the S-301 annual meetings to facilitate initiation of interagency collaborations. The value of attempting to interest ecologists and statisticians in the S-301 meeting to initiate future collaborations was also discussed. The difficulties of formulation research due to issues of proprietary information and patents belonging to biopesticide companies were briefly deliberated. It was generally agreed that collaborations in this area may still be productive, especially in the basic research arena. Brief research reports were given by attendees.

    3. Subproject 2: Development, evaluation and safety of entomopathogens for control of homopteran and other piercing-sucking insects (Rosalind James, Chair). Fungal pathogens are being developed as microbial control agents for the varroa mite (Varroa destructor), a pest in bee hives. Lygus in cotton was surveyed for naturally occurring fungal pathogens in California. Microbial control is being pursued as an option for controlling lygus in reservoir areas such as alfalfa, weeds, and safflower, rather than directly in cotton. Four entomopathogenic fungi have been isolated from the Chinese aphid, a new pest in the Midwest region. Neozygites is being developed for aphid control in California cotton, and Verticillium spp. for control the green peach aphid in dryland potatoes and for cabbage root aphid in cole crops. It was reported that Beauveria brognartii is now available commercially in Japan for control of stinkbugs on tree fruits, such as pear and apples. Thriponema is a small nematode that is an obligate parasite of thrips. It attacks thrips on foliage. Work is being done on the population structure of thrips and Thriponema using genetic markers. Beauveria bassiana is also being pursued as a possible microbial control for thrips. Works is now being done to determine whether Beauveria spores can attach to thrips from plant foliage. Approximately 200 strains of Hirsutella thompsonii are now in a culture collection at the University of Florida, Gainesville. Investigations are underway to determine the host range of these strains, in particular, the ability to infect both eriophyid and tetranychid mites.

    4. Subproject 3. Development, evaluation and safety of entomopathogens used in cryptic and soil habitats. (Co-Chairs Ed Lewis and Parwinder Grewal). A suggestion was made to develop a system for providing common names for entomopathogenic nematode species. The justification for this was the difficulty in using the long and difficult scientific names (e.g., Steinernema carpocapsae) during grower meetings for extension personnel. Generally, there was agreement that common names would be a good idea, but after considerable discussion, no consensus was reached on a systematic way to assign them. The value of long-term research on the effects of pathogens applied to soil ecosystems was discussed at length, citing such a study underway at NC State University as an example. In a similar vein, the off-target effects of entomopathogenic nematodes was discussed. Several members of this sub-project are involved in a USDA-funded research project that examines the interaction between entomopathogenic nematodes and several species of plant-parasitic nematodes. An update on the progress of that project was provided. Following sub-sub projects were identified in which there are established collaborations among different states:

      i. Non-target effects of entomopathogens: VA, NC, OH, CA, GA, NJ

      ii. Development of application techniques for entomopathogens: GA, VA, IL, NJ

      iii. Evaluation and Implementation of entomopathogens:

      Turfgrass: NJ, OH, VA, FL, (SDS-Japan)

      Nurseries: FL, OH, VA, OR

      Fruit crops: WA, FL, MD,

      Vegetables: OH, VA, WA

      Post harvest: CA, IL,

    5. Subproject 4. Development, evaluation and safety of entomopathogens for control of veterinary and structural arthropod pests (Co-Chairs James J. Becnel and David Oi). The first half of the session was devoted to structural pests. The multi-state collaborative project on microbial control of the imported fire ant involving members from 10 southern states [AL, AR, FL, GA, LA, MS, NC, OK, SC, & TN] was discussed. Members attending (Univ. TN, LSU, ARS Gainesville) discussed field inoculation studies with Thelohania solenopsae and new information on the life cycle that may help with the implementation of this control agent. A project investigating pathogens of termites involving the University of Florida and ARS Gainesville was discussed. The second half of the session was devoted to medical and veterinary pests. A collaborative project between Connecticut AES, California AES and ARS Gainesville was discussed with the objective to evaluate microbial agents (BTI and baculoviruses) for control of mosquitoes that vector West Nile Virus in the Northeast US. A collaborative project between CT AES and ARS Gainesville investigating phylogenic relationships of microsporidia in mosquitoes will be completed in 2001. A new microbial organism, Helicosporidia, was reported by investigators from ARS Gainesville and AES Florida who are involved in a project to conduct morphological and molecular characterization.

    6. The Local Arrangements Chair, Harry Kaya reported 50 registered attendees at this year?s meeting.

    7. The Site Selection Committee (Roberto Pereira, Chair) presented Orlando, FL, and Raleigh, NC as possible 2002 meeting sites. Raleigh was selected with Drs. Mary Barbercheck and James Harper as the Local Arrangement Co-Chairs. Tentative dates for the meeting are Feb. 24-25, 2002.

    8. Members were requested to consider seminar speakers for next years meeting. No travel funds are available for invited speakers, therefore potential speakers should be within the vicinity of Raleigh or be members attending the meeting.

    9. The format of the meeting was discussed. There was general agreement to continue with the 2001 format with no concurrent sessions.

    10. The Chair closed the meeting at 3:00PM.

    11. Members were invited to tour Agraquest?s research facility in Davis.

    Accomplishments:
    This project is a critical part of biological control and integrated pest management in the Southern Region. The current project involves insect pathologists, entomologists, microbiologists, molecular biologists, biochemists, geneticists, bacteriologists, mycologists, nematologists and vector biologists from 21 State Agricultural Experiment Stations, 12 USDA/ARS laboratories and 2 USDA/FS laboratories throughout the US and Puerto Rico. Participation also includes scientists from commercial industries involved in agricultural biotechnology and biological control.


    Substantial and important progress was made in the development and integration of entomopathogens into IPM systems.. Excellent progress was made in isolating and assessing new bacteria, fungi, microsporidia, nematodes and viruses providing new options and opportunities for control of lepidopteran, coleopteran and orthopteran pests of 1) crops (corn, cotton, soybeans, potatoes and other high value crops), 2) forests and 3) rangelands (Subproject 1). Considerable progress and advances were also made for control of homopteran and other piercing-sucking insect pests of cotton, ornamentals, vegetables, and greenhouse crops including: 1) mass culture and formulations of fungi 2) effectiveness of fungi for control of Varroa destructor, a honeybee parasite that causes serious economic damage to the bee keeping industry 3) integrating microbial control with other pest control measures and devising strategies for field and greenhouse use of microbial pesticides and 4) delivery systems for application of fungi (Subproject 2). Entomopathogenic fungi, nematodes and bacteria were evaluated for control of pests in cryptic and soil habitats (Subproject 3). Highlights include: 1) new isolates of BT were evaluated against surgacane borer, sweetpotato weevil and boll weevil, 2) new fungi and nematodes were isolated for control of orchard pests, 3) new information on the synergistic activity of imidacloprid and nematodes for control of grubs, 4) isolation and evaluation of fungi and nematodes for the Asian longhorn beetle. Entomopathogens for veterinary and structural pests was focused mainly on control of fire ants and mosquitoes (Subproject 4). Ten southern states are involved in a collaborative project to initiate infections in fire ant populations with a microsporidium pathogen and in 2000 infections were initiated in Oklahoma and South Carolina. New constructs of Bacillus thuringiensis subsp. israelensis (Bti) and B. sphaericus (Bs) and a new baculovirus showed excellent activity against mosquitoes and should be useful for controlling nuisance and vector mosquitoes, including the vectors of West Nile Virus.


    Researchers will continue to develop and implement entomopathogen technology that will significantly improve food safety and affordability, reduce the transmission of animal disease, protect biodiversity, enhance water quality and preserve the environment. New species and isolates of entomopathogens will be isolated for control of noxious insect pests throughout the United States. As many of the pests species are regional problems, there will be an emphasis for continued collaborations between states to test the efficacy, persistence, resistance management and other parameters of entomopathogens and their hosts under different sets of environmental conditions. In addition, the continued introduction of invasive species requires that new entomopathogens be discovered and evaluated for control of these pests that are a threat to agriculture, wildlife and humans in the United States.

    Impact Statements:
    Last Modified: unknown

    Date of Annual Report: 04/29/2002

    Report Information:
  • Annual Meeting Dates: 02/24/02 to 02/25/02
  • Period the Report Covers: 01/2001 to 12/2001

    • Participants:

    URL: Copy of participant list
    Brief Summary of Minutes of Annual Meeting:
    The meeting was convened at 9:00 AM by Chairman James J. Becnel.

    Jim Harper reported on local arrangements. Registration fee was raised from $25 to $40 to cover cost of meeting rooms since minimum number of hotel rooms was not reached. Jim thanked Mary Barbercheck for her participation on the local arrangements and Maureen Newman for helping with the registration.

    Minutes from the 2001 annual meeting were accepted.

    Chairman, Becnel reported: The Chair reported on the website and thanked the IPM center for their support and for hosting the webpages. The Chair announced that Ms. Peg Rotstein would talk later about the website, and asked for any suggestions on new material that should be added to the website. The need for an updated mailing list was discussed so it could be added to the website. A list was circulated for participants to correct their names and addresses.

    Dr. Jim Harper and Local arrangement Committee were commended for the organization and preparations in hosting the 2002 Annual Meeting of S-301 in Raleigh, NC.

    Dr. David Boethel (SARES Administrative Advisor) presented his report to the members (attached) and congratulated participants for the extensive publication list, which makes this regional project one of the most productive.

    A site selection committee was formed with Member at Large, Roberto Pereira, chair, who invited Don Steinkraus to participate in selecting next site.

    Summary of the Discussion:

    Mary Barbercheck introduced the speaker, Dr. Fred Gould, North Carolina State University, who made a presentation titled Biological Control, Transgenic crops, and Insect Resistance.

    Peg Rotstein described S-301 website and demonstrated main features. Following her presentation, the attendees discussed several changes to the website including: a) new name for the site to be easier to be identified in Internet searches; b) addition of member information to the site (approved by the members presentat the meeting); c) addition of links page including possible links to electronic publications, the entomopathogenic nematode website, APHIS-PPQ site with forms and phone numbers.

    Subproject 1: Development, evaluation and safety of entomopathogens for leaf feeding insect defoliators (Co-Chairs: Stephen Wraignt and Leellen Solter). Studies at nine state and federal research institutions pursued development of numerous viral, bacterial, and fungal pathogens for microbial control of lepidopteran, coleopteran, and orthopteran pests, including diamondback moth, blueberry spanworm, beet armyworm, corn earworm, celery looper, soybean looper, Colorado potato beetle, boll weevil, sweet potato weevil, strawberry rootworm, and grasshoppers. Colorado potato beetle and the lepidopteran complex on cabbage were effectively controlled with integrated applications of the bacterium Bacillus thuringiensis and fungus Beauveria bassiana. Fundamental studies elucidated mechanisms of developmental resistance to nucleopolyhedroviruses (NPV) in corn earworm and tobacco budworm, population genetics of the fungus Nomuraea rileyi infecting green cloverworm and velvetbean caterpillar, and ascovirus-lepidopteran host-parastioid interactions. Soil to plant transport of NPV was measured and evaluated as a risk factor associated with field release of recombinant viruses. Comparisons of Bt cotton versus conventional cotton showed no observable negative effects of Bt cotton on non-target insect populations. Researchers at 11 U.S. state and federal research institutions and collaborating scientists from four nations conducted studies of fungal, microsporidian and viral pathogens for microbial control of lepidopteran pests of forestry, including gypsy moth, browntail moth, forest tent caterpillar, Douglas fir tussock moth, and Io moths. Basic studies investigated resting-spore dormancy and host range in the fungus Entomophaga maimaiga, DNA fingerprinting of microsporidian pathogens, DNA sequencing of several cypoviruses, identification of chemical enhancers of gypsy moth NPV, and gypsy moth-parasitoid-polydnavirus interactions. Improved methods were developed for production of E. maimaiga resting spores in the laboratory.

    Subproject 2: Development, evaluation and safety of entomopathogens for control of homopteran and other piercing-sucking insects (Rosalind James, Chair). Work has continued toward developing the use of entomopathogenic fungi as microbial controls for mites, aphids, whiteflies, and lygus bugs. This research involved a variety of agricultural systems, including honey bees, cotton, soybeans, and vegetable crops. This year, a considerable effort was put into gaining a better understanding of disease outbreaks in these insects. Beauveria bassiana was identified as a major pathogen of lygus bugs, and Neozygites fresenii was shown to be a natural control of cotton aphids for the 9th consecutive year. Hirsutella thompsonii was also investigated for its potential in mite control. A PCR marker was developed to test for the presence of a H. thompsonii toxin. This fungus was also found to have good activity against the varroa mite, which is a parasite of honey bees. In addition, a formulation was developed for fungal pesticides using encapsulation technology previously used for baculoviruses.

    Subproject 3. Development, evaluation and safety of entomopathogens used in cryptic and soil habitats. (Co-Chairs Ed Lewis and Parwinder Grewal). Work continued towards the discovery and identification of more virulent species and strains of entomopathogenic nematodes, bacteria, and fungi for the development of biological control for white grubs, weevils (boll weevil, carrot weevil, citrus weevil, plum curculio, pecan weevil, sweet potato weevil), fire ants, codling moth, Asian longhorn beetle, and termites. Photorhabdus bacteria were shown to be able to independently infect termites. Nematode strains with increased storage stability (longevity) and tolerance to major environmental stresses were identified and compatibility with chemical insecticides and formulation ingredients was studied. Research also continued on the mass production and delivery systems of nematodes. Addition of the wetting agent Silwet -L77 increases nematode efficacy and reduces toxicity of chlorine. Heterorhabditis bacteriophora survives better in muck soils than Steinernema carpocapsae. Also H. bacteriophora is more abundant than Steinerenema carpocapsae in physically disturbed soil systems. Entomopathogenic nematodes are less abundant in pasture rotation but entomopathogenic fungi were more abundant. It was demonstrated that nematodes can adapt to laboratory conditions that can produce dramatic changes in important biological traits. Both live and dead entomopathogenic nematodes suppress soil populations of many plant-parasitic nematodes especially in perennial crops (turfgrass and woody ornamentals), but they have no detrimental effects on free-living nematodes. It was also shown that the application of entomopathogenic nematodes has the strongest effect when they are applied at the same time with plant parasitic nematodes. Indianmeal moth granulosis virus was registered by the US EPA in December 2001 and has been licensed to Agrivir, LLC.

    Subproject 4. Development, evaluation and safety of entomopathogens for control of veterinary and structural arthropod pests (Co-Chairs James J. Becnel and David Oi). Field inoculations of the microsporidium Thelohania solenopsae into imported fire ants colonies in ten southern states has resulted in infection and spread predominately in the multiple-queen form of the red imported fire ant, Solenopsis invicta, and laboratory inoculations resulted in the demise of multiple-queen colonies. A new protozoan disease, tentatively identified as a Mattesia sp. was observed in S. invicta from Florida and soil types affected the efficacy of Beauveria bassiana on this ant. Early sporulation was more important than virulence in transmission and epizootics of Metarhizium anisopliae, which killed Formosan termites more effectively than B. bassiana. Entomopathogens evaluated against mosquitoes included viruses, microsporidia, mermithid nematodes and a new insect pathogenic algae. Basic studies focused on the taxonomy and life histories of new pathogens while other investigations focused on safety, efficacy against the target hosts and improving production and delivery systems of promising agents.

    Final Business Meeting

    The Site Selection Committee (Roberto Pereira, Chair) presented Charleston, SC as possible 2003 meeting site and Dr. Gerry Carner as Local Arrangements, with tentative meeting dates as Feb 23-24, 2003. Gerry said a few words about the location. Gainesville, FL, was presented by Roberto Pereira and Jimmy Becnel as possible 2004 meeting location. Meeting locations were approved by attending members.

    Nominating Committee (Ted Andreadis, Chair) presented Parwinder Grewal as candidate for Member-at-Large position. No other nominations were presented and Harry Kaya moved that nominations be closed. By unanimous vote, P. Grewal was elected as new Member-at-Large.

    Members were requested to consider seminar speakers for next years meeting. No travel funds are available for invited speakers, therefore potential speakers should be within the vicinity of Charleston, SC or be members attending the meeting. Suggestions for topic included regulatory aspects and interaction between biological control and ecology. Several members stated that the regulatory aspects had already been discussed in previous meetings.

    The Chair closed the meeting at 3:00PM.

    John Vanderberg thanked Jimmy Becnel, Chair for the past 2 years, for his dedication and hard work.

    Next Meeting Information:

    Location and Date: Charleston, SC, February 23-24, 2003

    Responsible Individual: Gerry Carner

    Between Meeting Information Exchange/Development Information:

    Accomplishments:
    This project is a critical part of biological control and integrated pest management in the Southern Region. The current project involves insect pathologists, entomologists, microbiologists, molecular biologists, biochemists, geneticists, bacteriologists, mycologists, nematologists and vector biologists from 21 State Agricultural Experiment Stations, 12 USDA/ARS laboratories and 2 USDA/FS laboratories throughout the US and Puerto Rico. Participation also includes scientists from commercial industries involved in agricultural biotechnology and biological control. The project is composed of the following 4 subprojects followed by a summary of the years activities. The complete annual report with publications can be found at the S-301 website http://cipmtest.ent.ncsu.edu/s301/.

    SUBPROJECT 1: Development, evaluation and safety of entomopathogens for leaf feeding insect defoliators. Studies at nine state and federal research institutions pursued development of numerous viral, bacterial, and fungal pathogens for microbial control of lepidopteran, coleopteran, and orthopteran pests, including diamondback moth, blueberry spanworm, beet armyworm, corn earworm, celery looper, soybean looper, Colorado potato beetle, boll weevil, sweet potato weevil, strawberry rootworm, and grasshoppers. Colorado potato beetle and the lepidopteran complex on cabbage were effectively controlled with integrated applications of the bacterium Bacillus thuringiensis and fungus Beauveria bassiana. Fundamental studies elucidated mechanisms of developmental resistance to nucleopolyhedroviruses (NPV) in corn earworm and tobacco budworm, population genetics of the fungus Nomuraea rileyi infecting green cloverworm and velvetbean caterpillar, and ascovirus-lepidopteran host-parastioid interactions. Soil to plant transport of NPV was measured and evaluated as a risk factor associated with field release of recombinant viruses. Comparisons of Bt cotton versus conventional cotton showed no observable negative effects of Bt cotton on non-target insect populations. Researchers at 11 U.S. state and federal research institutions and collaborating scientists from four nations conducted studies of fungal, microsporidian and viral pathogens for microbial control of lepidopteran pests of forestry, including gypsy moth, browntail moth, forest tent caterpillar, Douglas fir tussock moth, and Io moths. Basic studies investigated resting-spore dormancy and host range in the fungus Entomophaga maimaiga, DNA fingerprinting of microsporidian pathogens, DNA sequencing of several cypoviruses, identification of chemical enhancers of gypsy moth NPV, and gypsy moth-parasitoid-polydnavirus interactions. Improved methods were developed for production of E. maimaiga resting spores in the laboratory.

    SUBPROJECT 2. Development, evaluation and safety of entomopathogens for control of homopteran and other piercing-sucking insects. Work has continued toward developing the use of entomopathogenic fungi as microbial controls for mites, aphids, whiteflies, and lygus bugs. This research involved a variety of agricultural systems, including honey bees, cotton, soybeans, and vegetable crops. This year, a considerable effort was put into gaining a better understanding of disease outbreaks in these insects. Beauveria bassiana was identified as a major pathogen of lygus bugs, and Neozygites fresenii was shown to be a natural control of cotton aphids for the 9th consecutive year. Hirsutella thompsonii was also investigated for its potential in mite control. A PCR marker was developed to test for the presence of a H. thompsonii toxin. This fungus was also found to have good activity against the varroa mite, which is a parasite of honeybees. In addition, a formulation was developed for fungal pesticides using encapsulation technology previously used for baculoviruses.

    SUBPROJECT 3: Development, evaluation and safety of entomopathogens in cryptic and soil habitats. Work continued towards the discovery and identification of more virulent species and strains of entomopathogenic nematodes, bacteria, and fungi for the development of biological control for white grubs, weevils (boll weevil, carrot weevil, citrus weevil, plum curculio, pecan weevil, sweet potato weevil), fire ants, codling moth, Asian longhorn beetle, and termites. Photorhabdus bacteria were shown to be able to independently infect termites. Nematode strains with increased storage stability (longevity) and tolerance to major environmental stresses were identified and compatibility with chemical insecticides and formulation ingredients was studied. Research also continued on the mass production and delivery systems of nematodes. Addition of the wetting agent Silwet -L77 increases nematode efficacy and reduces toxicity of chlorine. Heterorhabditis bacteriophora survives better in muck soils than Steinernema carpocapsae. Also H. bacteriophora is more abundant than Steinerenema carpocapsae in physically disturbed soil systems. Entomopathogenic nematodes are less abundant in pasture rotation but entomopathogenic fungi were more abundant. It was demonstrated that nematodes can adapt to laboratory conditions that can produce dramatic changes in important biological traits. Both live and dead entomopathogenic nematodes suppress soil populations of many plant-parasitic nematodes especially in perennial crops (turfgrass and woody ornamentals), but they have no detrimental effects on free-living nematodes. It was also shown that the application of entomopathogenic nematodes has the strongest effect when they are applied at the same time with plant parasitic nematodes. Indianmeal moth granulosis virus was registered by the US EPA in December 2001 and has been licensed to Agrivir, LLC.

    SUBPROJECT 4: Development, evaluation and safety of entomopathogens for veterinary and structural arthropod pests. Field inoculations of the microsporidium Thelohania solenopsae into imported fire ants colonies in ten southern states has resulted in infection and spread predominately in the multiple-queen form of the red imported fire ant, Solenopsis invicta, and laboratory inoculations resulted in the demise of multiple-queen colonies. A new protozoan disease, tentatively identified as a Mattesia sp. was observed in S. invicta from Florida and soil types affected the efficacy of Beauveria bassiana on this ant. Early sporulation was more important than virulence in transmission and epizootics of Metarhizium anisopliae, which killed Formosan termites more effectively than B. bassiana.

    Entomopathogens evaluated against mosquitoes included viruses, microsporidia, mermithid nematodes and a new insect pathogenic algae. Basic studies focused on the taxonomy and life histories of new pathogens while other investigations focused on safety, efficacy against the target hosts and improving production and delivery systems of promising agents.



    Impact Statements:
    1. This project impacts biological control and integrated pest management in the Southern Region through the development and integration of entomopathogens into IPM systems Progress was made in isolating and assessing new bacteria, fungi, microsporidia, nematodes and viruses for control of lepidopteran, homopteran, coleopteran, dipteran, and orthopteran pests of crops (corn, cotton, soybeans, potatoes and other high value crops), forests, rangelands, and man and animals.
    Last Modified: unknown

    Date of Annual Report: 04/01/2003

    Report Information:
  • Annual Meeting Dates: 02/23/03 to 02/24/03
  • Period the Report Covers: 01/2002 to 12/2002

    • Participants:
    Brief Summary of Minutes of Annual Meeting:
    The meeting was convened at 9:15 am by Chair E. Groden. Gerry Carner reported on local arrangements. Registration fee was set at $35 because only 30 participants had registered by the start of the meeting. Gerry reminded participants that Charleston was voted the most polite city in US, recommended using the shuttle to go downtown, and recommended a few attractions. Jim
    thanked M. Sheppard and D. Palmer for helping local arrangements and his student for helping with the registration. Minutes from the 2002 annual meeting. Since these have been sent to all participants and are posted on website. Jim Becnel moved that minutes be accepted without reading. The motion was seconded by Ed Lewis and the 2002 minutes were accepted unanimously. Chair Groden reported: The chair thanked subproject chairs and members for
    report submissions. The need for updated list of participants was raised and a list was passed around so members could make necessary changes. The Chair thanked Ms. Peg Rotstein for her work maintaining the website and mentioned that the participants may want to update and enhance the website, especially due to the upcoming project renewal. Dr. Groden asked for ideas
    from the participants. The chair introduced Dr. David Boethel, and asked him to talk about steps involved in renewing the project. Dr. David Boethel (SARES Administrative Advisor) thanked E. Groden and G. Carner for organizing the meeting. He commended the participants for having the report done at the time of the meeting, having a good record of publications, and commented
    on the new reporting system that will be online soon, after taking care of few glitches. D. Boethel recommended that participants write brief and to-the-point reports, concentrating on impact to stakeholders. He commented on the need to update the participant list, but that this will be simplified by the new reporting system. New participants in the project, whether from state experimental stations or from USDA need to prepare Appendix E and submit it through the Exp. Station Directors Office. As for the project rewrite, D. Boethel recommended: Identifying the people that will be involved in the rewrite. Using the next year to start the rewriting process. B. Federici asked how many people were currently participating in the project. E. Groden said that participating list had to be updated, but that the mailing list included more than 80 names. According to D. Boethel, other multi-state projects are in general not as large as S-301, but that these projects are getting larger, and becoming more national rather than regional, as in the past. Asked (by C. McCoy) about how money for these multi-state projects go to the states, Boethel responded that a formula based on population and number of farms determines how much money goes to the states, and multi-state project money has to be used for this purpose only. However, use may not be apparent for state researchers and may cover travel costs, portion of salaries, etc. Each state handles the multi-state project money differently. E. Groden instructed subproject chairs to discuss project renewal in separate meetings, including discussions on how participants feel about structural changes made in the last renewal. By next years meeting we should have the objectives for the new project. Next years meeting is scheduled to be in Gainesville, FL B. Federici suggested Laguna Beach for the 2005 meeting, and M. McGuire offered to help with organization. Each of the participants introduced him/herself and said a few words about the research conducted in relation to the multi-state project. Bob Nowierski the USDA-CSREES, Plant and Animal System representative made a short presentation on CSREES and its grant programs. B. Nowierski is the National Program Leader for Bio-based Pest Management. He discussed several different programs and new initiatives within CSREES that relate to the S-301 project. He asked participants whether there was adequate funding for biopesticide research, especially in the area of development and extension. He announced the 4th National IPM Symposium on April 8-10, 2003 in Indianapolis, and discussed several grant opportunities. Bob also listed the CSREES priorities for 2003-2005 as: agricultural security, obesity, human nutrition and food science, food safety, genomics, future food and fiber production quality, agricultural and environmental quality, sustainable agriculture to produce rural and community prosperity. E. Groden introduced the speaker, Dr. Parwinder Grewal, Ohio State University, who made a presentation tritrophic interactions in microbial control concentrating on the effects of plant endophytes on microbial control agents used in pest management. During the Subproject 3 meeting, the participants discussed the rewrite of the S-301 Project. Participants need to identify milestones and potential outcomes for the project. Some potential objectives discussed by the participants were: invasive species; control of diseases in beneficial insects; production and formulation of entomopathogens; diagnostics and biosafety. Nowierski suggested using Goals in the project rewrite to designate long-term objectives and Objectives for accomplishable short-term objectives that can be measured within the project life. Other suggestions were: emphasize multi-state cooperation; use matrixs to demonstrate who works on which project and show integration and multi-state characteristics; within each subproject, some objectives to use can be: identification of new pathogens; production and formulation; practical use of pathogens; non-target effects; invite industry people back into the project. Future seminar topics were discussed including: future of microbial control industry; GMOs as future of insect/disease control; microbial control in nature (classical biological control using pathogens). Mary Barbercheck suggested a possible grant dealing with post-mortem dynamics of entomopathogenic microorganisms including predation, movement of cadavers, decomposition and other issues. Each of the subprojects met and discussed research progress, collaborations and other issues related to the subproject. Subproject 1: development, evaluation and safety of entomopathogens for leaf feeding insect defoliators (Chair: Bob Bailey). Subproject 2: Development, evaluation and safety of entomopathogens for control of homopteran and other piercing-sucking insects (Chair: Rosalind James, for Michael Brownbridge who could not attend the meeting). Subproject 3: Development, evaluation and safety of entomopathogens used in cryptic and soil habitats. (Chair: Ed Lewis). Subproject 4: Development, evaluation and safety of
    entomopathogens for control of veterinary and structural arthropod pests (Chair: James Becnel). A rewrite committee was nominated including all the project officials and the subproject chairs. Report needs a section on impacts so members should submit impact statements (bullets) to the subproject chairs. The Site Selection Committee Chair Parwinder Grewal asked Roberto Pereira
    to present report. Besides the Laguna Beach suggestion made by Brian Federici at the beginning of the meeting, Danny Brook offered to host the 2005 meeting in either Corvalis or Portland, Oregon. The offer was accepted by the members present at the final meeting. The current format with meetings on Sunday and Monday will be voted on by e-mail. Some members felt that the current format may have caused less people to come to the meeting. Next meeting is currently
    scheduled for Gainesville, FL, to be organized by Jim Becnel, Drion Boucias and Roberto Pereira, possibly for the period between February 20 and 28, 2004. A suggestion to hold a stackholder meeting in connection to the meeting in Gainesville was well received by the members. The IPM Center in FL, directed by Russ Mizzel, was suggested as possible funding source to bring stakeholder participants. Some discussion on the meeting with possibility of invited speaker (Doug Streett) to talk about USDA facility in Mississippi for biocontrol mass-production. Roberto Pereira asked project members to sign as SIP members and encourage others that may work in the area of insect pathology and microbial control to do the same. David Oi and Ann Hajek were suggested as speakers for seminar for next years meeting, concentrating on classical biological control of insects using microorganisms.

    Additional information can be viewed at the following address: http://cipm5.ent.ncsu.edu/s301/index.cfm

    Accomplishments:
    Subproject 1. Development, evaluation and safety of entomopathogens for leaf feeding insect defoliators.

    Ten state agencies and eight USDA field stations reported studies on the development of pathogens, including fungi, baculoviruses, bacteria, and microsporidia, for biological control against the following leaf-feeding hosts in agricultural and forest systems: bollworm, browntail moth, cabbage loopers, cassava green mite, codling moth, Colorado potato beetle, diamondback
    moth, European sawfly, fall armyworm and other Spodoptera species, grasshoppers, green cloverworm, gypsy moth, Pandemis leafroller, and velvetbean caterpillar. Several taxonomic, host specificity, and competition studies are in progress to evaluate three genera of gypsy moth and browntail moth microsporidia as possible classical biological control agents of the two oak
    defoliators in the U.S. Studies of the environmental reservoirs and efficacy of the fungal pathogen Entomophaga maimaiga continue due to the demand for the pathogen as a control agent of the gypsy moth. An SNPV of the European sawfly was found to be distantly related to lepidopteran nucleopolyhedrosis viruses. Commercial formulations (Carpovirusine and Cyd-X) of the codling moth granulovirus provided significant reduction of deep entries to fruit and
    reduction of live larvae. Nucleopolyhedrosis viruses are being evaluated for several field crop defoliators. Bt-transgenic corn and SfNPV appear to have additive effects on fall armyworm, and LdNPV was successfully tested as a synergist for other NPV isolates against noctuid pests. In addition, soil to plant transport of NPV occlusion bodies was evaluated under variable
    environmental conditions and soil depth. Storage time of AfMNPV was found to be more than 30 months under refrigeration, but storage time declined significantly at 30 C. Ascoviruses collected from noctuid pests in SC are being compared to Indonesian isolates. Several projects utilizing entomopathogenic fungi against field crop pests were reported. Beauveria bassiana in canola and corn oil carriers increased mortality of grasshoppers over that of paraffinic oil carriers and factors involved in the secondary cycling of B. bassiana are under evaluation, as well as the impact of commercial B. bassiana products on native strains. A spray-dried encapsulation of B. bassiana retained initial viability but resistance to UV radiation was not improved. Taxonomic differences between South American and African Neozygites infecting green cassava mites are being determined, and the effects of drought on Nomuraea rileyi infections of soybean pests are being studied. No negative effects on nontarget hosts were found in studies on Bt cotton.

    Subproject 2. Development, evaluation and safety of entomopathogens for homopteran and other piercing-sucking insects.

    Entomogenous fungi are being developed for the microbial control of mites, aphids, thrips, stink bugs and lygus bugs. This research is being done across a diverse range of agricultural crops, including cotton, soybeans, rice, vegetable crops and ornamentals, as well as for control of honey
    bee parasites. This year, considerable effort was put into gaining a better understanding of disease outbreaks in these insects, and the incidence of entomopathogens in pest populations. Beauveria bassiana was identified as a major natural pathogen of lygus bugs and was the predominant fungal species recovered from avocado thrips habitats in CA. Neozygites fresenii-
    induced epizootics occurred in cotton aphid populations for the 10th consecutive year, and appeared to be compatible with agropesticides used in integrated control programs for this pest. Metabolites produced by sporulating cultures of Hirsutella thompsonii inhibited oviposition in two-spotted spider mite and shows promise for further development as a control agent for this
    pest. In field trials, Metarhizium anisopliae was shown to be effective against the parasitic varroa mite, and had minimal to no harmful side-effects on bee brood survival or honey production. Trials are continuing on ways of improving fungal efficacy against thrips through use with selected biorational insecticides and improving spray application techniques. B. bassiana and M. anisopliae were tested in small-scale field trials to control lygus bugs on lettuce but results were inconsistent, and generally low levels of infection/control were obtained, probably as a result of the hot, dry test conditions.

    Subproject 3. Development, evaluation and safety of entomopathogens in cryptic and soil habitats.

    Eleven state agencies and seven USDA field stations submitted research results for this report. Reported work includes several different types of studies that include all five groups of insect pathogens. A combination of near-term and long-term results are included. Field and laboratory trials of various pathogen-based pest management materials that are either available commercially or at some stage of development were tested against 18 different species of arthropod pests, 4 species of plant-parasitic nematode pests and one species of slug pest. These trails were conducted in 14 different commodities including food and horticultural crops and structural resources. Two pathogen surveys were reported, one of which yielded 10 new isolates of the entomopathogenic nematode Steinernema riobrave. Four studies of non-target impacts of insect pathogen releases were conducted. Three of these studies were with entomopathogenic nematodes; one to examine the interaction between entomopathogenic nematodes and ichneumon parasitoids, one to determine the effects of entomopathogenic nematodes on plant-parasitic nematodes and one to record interactions between applied entomopathogenic nematodes and soil fauna in general. The fourth study aimed to determine the effects of metabolites of Photorhabdus bacteria against plant-parasitic nematodes. Studies of various traits of entomopathogenic nematodes entomopathogenic fungi maintained in culture were also reported.

    Subproject 4. Development, evaluation and safety of entomopathogens for veterinary and structural arthropod pests.

    Entomopathogenic research on structural pests focused on the control of the red imported fire ant, Solenopsis invicta, and termites. Utilization of the microsporidium Thelohania solenopsae, is currently being evaluated in the field as a control for fire ants either by itself or as part of an integrated approach combining biological and chemical controls. Improved T. solenopsae
    detection protocols have been developed utilizing either PCR or histological stains. Other studies have examined intercolony transmission and potentiation of insecticides in infected ants. Beauveria bassiana isolates varied in their control of fire ants. Development cycles of the fungi Beauveria bassiana and Metarhizium anisopliae were compared in termites. Entomopathogens evaluated against mosquitoes included bacteria, viruses, fungi, microsporidia,
    and a new insect pathogenic algae. A novel recombinant Bacillus thuringiensis subsp. israelensis (Bti) strain and the baculovirus CuniNPV have shown excellent activity against Culex mosquitoes that vector West Nile Virus in the U.S. Basic studies focused on the taxonomy and life histories of new pathogens while other investigations focused on safety, efficacy against the target hosts and improving production and delivery systems of promising agents.

    Impact Statements:
    1. Project impacts biological control and IPM in the Southern Region through the development and integration of entomopathogens into IPM systems.
    2. New bio-pesticides for the control of mosquitos that vector diseases in the U.S. were discovered.
    3. Fire ant colonies were infected with Thelohania solenopsae and spread of the pathogen was documented.
    4. New entomopathogens were isolated and assessed for control of major insect pests.
    5. New technologies discovered provide safe and effective control with reduced pesticide load to the environment.
    Last Modified: unknown

    Date of Annual Report: 04/13/2004

    Report Information:
  • Annual Meeting Dates: 02/22/04 to 02/23/04
  • Period the Report Covers: 01/2003 to 12/2003

    • Participants:
    Brief Summary of Minutes of Annual Meeting:
    MINUTES OF THE 2004 S-301 TECHNICAL COMMITTEE MEETING

    Location and Date of the Meeting: Entomology & Nematology Department, University of Florida and USDA-ARS, Center for Medical, Agricultural and Veterinary Entomology, Gainesville, FL; Feb. 22-23, 2004.

    Action Items/Assigned Responsibilities/Target Dates:

    1. The meeting was convened at 9:10 AM by Chair E. Groden, with attendance of 31 participants.

    2. Roberto Pereira, Jim Maruniak and Jim Becnel reported on local arrangements. Registration fee was set at $30. J. Maruniak explained aspects of the Entomology & Nematology Department. J. Becnel explained that the Tuesday morning meeting would convene at the USDA-ARS lab where name badges would be necessary due to security concerns.

    3. Minutes from the 2003 annual meeting. Since these have been sent to all participants, S. Jaronski moved that minutes be accepted without reading. The motion was seconded by V. D. Amico and the 2003 minutes were accepted unanimously.

    4. Chair Groden reported:

    The chair thanked subproject chairs and members for report submissions.

    The chair talked about the development of the pre-proposal for project renewal. This process is done electronically now utilizing a short form. E. Groden instructed the subcommittees to discuss proposal renewal including the highlighting of accomplishments and listing of interested stakeholders.

    For the proposal, the methods section must indicate what the different members want to do in terms of research. Any changes from past proposal must be reported to the subcommittee chairs so changes are made.

    The Chair asked P. Grewal (Member-at-Large) to coordinate the selections of next years meeting location, and announced that L. Lacey would be talking on Tuesday on behalf of Denny Bruck from USDA-ARS lab in Corvallis ,OR, on arrangements for the 2005 meeting.

    The chair introduced Dr. David Boethel, and asked him to talk about administrative aspects of the project.

    5. Dr. David Boethel (SARES Administrative Advisor) commended the participants for being on time for the renewal process and recommended the formation of a committee to deal with project renewal. He recommended that the proposal be ready by fall 2004 for the September Exp. Station Directors meeting. He recommended that names of non-members be submitted as possible reviewers for the project. The project goes also to Department heads for review.

    D. Boethel mentioned that the minutes and annual report should be ready 60 days before the annual meeting.

    He thanked the S-301 leadership for organizing the meeting and putting the project together.

    6. Rick Meyer, attending the meeting for Bob Nowierski, the USDA-CSREES, Plant and Animal System representative, distributed a handout with information on budget and other aspects of the CSREES . He commented that the budget has changed because of many other issues affecting the federal government , specifically Homeland Security and the current military actions. There is a 10% cut in the CSREES programs, but some increases in Homeland Security may also benefit some of the CSREES programs that relate to that subject. Competitive grant programs were cut but not greatly.

    The 2005 budget proposal includes some increases for NRI grant program. In this program, 20% of the budget has to be used for integrated Research/Extension/Education programs. The organic transitions program would get a good increase in funds, and a new Request for Proposals (RFP) should be out soon for this grant program.

    Some of the subjects that B. Nowierski wanted to comment on included:
    - The USDA has been working on resolving the issue of requirement by the USDA-APHIS for a bonded carrier for hand carrying biological control agents through customs as they enter the US.
    - The Denver meeting on biological control identified some areas that need further attention such as: a) better prediction of biocontrol effects, b) regulatory reform, and c) risk analysis.

    R. Meyer also commented on the USDA-ARS Biological Control Workshop that many of the ARS scientists attended in early February 2004.

    7. J. Becnel, coordinator for this USDA-ARS Biological Control Workshop commented on the meeting. The workshop served to identify several critical issues in biological control of weeds, insects, and plant pathogens. Approximately 120-130 ARS scientists participated and addressed impediments and issues important to biological control. First, there were sections looking at broad issues in inter-disciplinary groups, and then discussions within each discipline. There were 12 task teams formed that will be looking at several identified issues.

    8. Rick Meyer mentioned that members should keep in mind while developing the new project that the USDA uses a performance-based assessment of projects. The project should have links to the USDA goals and propose outcomes and outline past and future impacts. The project should not simply list outcomes (publications) but should contain outcomes that benefit the stakeholders.

    9. The discussion was followed by the plenary section where each of the participants introduced him/herself and said a few words about the research conducted in relation to the multi-state project.

    10. The Plenary section was followed by lunch and then the presentation of 2 seminars by: Dr. Ann Hajek, Classical Biological Control Using Entomopathogenic Fungi, and Dr. David Oi, Thelohania solenopsae as Classical Biological Control for Imported Fire Ants.

    11. During this years meeting, the subcommittee meetings were dominated by discussions on the renewal of the project proposal. Discussions included the need for:
    - a Strategic Plan for the use of microbials in each of the subobjective
    - the inclusion of a workshop on each of the subobjectives (during meetings in consecutive years).
    - A restructuring of the subobjectives under a different organization that better reflects a preoccupation with the impacts of the research done under this project.

    12. The meeting format also was discussed with a proposal to eliminate the plenary section and combine it with the seminar in a workshop, including not only presentations by leading experts but also a discussion including invited stakeholders (farmers, regulators, etc). As a final product, a document including conclusions and recommendations from the workshop would be produced and posted in the project webpage. These recommendations could serve as the basis for future research as well as policy changes and other governmental actions.

    13. There is a need to get more industry people involved in the project. Attendance of these industry representatives has declined in recent years due to decline in number of industries involved in microbial control and the decline of interest in the meeting. E. Groden and project administrators commented that there is no problem with the size of the group, which is larger than most other similar multi-state projects.

    14. D. Boethel commented that the S-301 exists to facilitate collaboration. R. Meyer added that that collaboration must be clearly demonstrated in the project proposal and the reports. S. Jaronski commented that other groups have a more obvious collaboration with large experiments conducted at different states simultaneously under the supervision of several cooperators.

    15. L. Tedders commented on the void between industry and research and that something should be done to fill that gap. P. Grewal commented that past format for the S-301 meeting fostered more communication with the industry representatives because subobjectives were the different groups of pathogens. D. Shapiro commented that in the past people discussed their research in more details in the subobjective meetings. R. Pereira suggested that the present format is OK but that more discussion is needed. M. Brownbridge agreed more interactions between participants was needed.

    16. E. Groden commented on the need to balance the number of people interested in each subobjective in order to have a better meeting. P. Grewal suggested a restructuring of the project and the meeting with a commodity focus. He commented that most of the funding is commodity oriented and that would facilitate grant proposals from the project participants. D. Boethel and R. Meyer agreed that that would be a good idea.

    17. The participants voted to restructure the project with a commodity focus. Several different groups were suggested and discussed. The groups were then reorganized and voted on with a final decision of 4 new subobjectives for the renewal of the S-301 project. These are:
    - Row Crops pests, which will have R. Behle as the subobjective chair.
    - Medical, Veterinary and structural pests, which will have D. Oi and J. Becnel as the subobjective chairs.
    - Horticulture pests, including vegetables, greenhouse, and fruits and nuts, which will have M. Brownbridge and D. Shapiro-Ilan as the subobjective chairs.
    - Urban Landscape, Ornamentals and Forest pests, which will have E. Lewis and P. Grewal as the subobjective chairs.

    18. J. Becnel proposed that within each subobjective the research would be divided into: a) discovery, b) integration, c) implementation, and d) environmental safety.

    19. The new project format was approved by 21 votes, opposed by 1 vote, and there were 3 abstentions.

    20. D. Boethel said that the preproposal is just 3 pages long and does not include much in terms of procedures. The deadline the project leaders should work with is Fall 2004 (third week in September is when the committee will be meeting to discuss the renewal). So the project should be in D. Boethels hands by August 2004. For the project, participants need to contribute by sending:
    - Accomplishments
    - Time commitment in terms of Scientists Years (SY) and Technician Years (TY)
    - Highlighted cooperative publications (Multi-institution publications)

    21. L. Lacey made presentation sent by D. Bruck on the tentative meeting venue for 2005. Tentative dates for that meeting were set at Sunday and Monday February 27-28, 2005. A discussion on the packages offered by the hotel followed but the participants agreed that getting the package would be hard due to the need for an advance payment.

    22. The meeting format was then discussed. The format proposed earlier in the meeting was adopted. The organizers will invite stakeholders and experts to participate in workshop discussion on the use of microbials on fruit crops. The discussion will lead to the development of recommendations for future research areas.

    23. The election of a new member-at-large was the next item in the agenda. Ed Lewis accepted his nominations and was elected by acclamation . The meeting was adjourned in the afternoon of February 23, 2004.

    Next Meeting Information: February 27-28, 2005.

    Location and Date: Portland area, OR

    Responsible Individuals: Danny Brook and Larry Lacey

    Accomplishments:
    SUBPROJECT 1. Development, evaluation and safety of entomopathogens for leaf feeding insect defoliators.

    Coordinator: Robert Behle, USDA,ARS-Peoria
    Cooperating Agencies: AL, CT, FL, IL, IL-INHS, LA, ME, NY, SC, WY; ARS-Beltsville, Fresno, Ithaca, Peoria, Sidney; USDA/FS- CT, MI; USDA APHIS
    Reporting: R. Behle, D. Boucias, G. Carner, R. Farrar, J. Fuxa, E. Groden, A. Hajek, S. Jaronski, W. Moar, L. Solter

    Summary

    Ten scientists from seven state agencies and three USDA research groups reported studies concerning the use of entomopathogens for control of leaf-feeding insects. Fewer reports were received when compared with past years and represents a small decline in the research directed at leaf feeding insects as some previous contributors have shifted efforts to cryptic feeding insects. Targeted leaf-feeding insect pests represented three insect orders: coleoptera (Mexican bean beetle, Colorado potato beetle), lepidoptera (corn earworm, diamondback moth, fall armyworm, green cloverworm, beet armyworm, and velvetbean caterpillar, gyspy moth, Eastern tent caterpillar), and orthoptera (American grasshopper). A wide variety of pathogens continue to be evaluated including (but not limited to): baculovirus, fungus and protozoa (microsporidia) with some additional research reported on Bt-transgenic plants. Application environments ranged widely from perennial systems such as range grass, forests and urban landscapes to annual crops such as sweet corn, cotton, soybeans, potatoes and vegetable crops. As a result of the research conducted by this group and collaborators, twenty-seven manuscripts in various phases of publication are listed.


    Sub-project 2: Development, evaluation and safety of entomopathogens for homopteran and other piercing-sucking insects.

    Coordinator: Michael Brownbridge, Univ of VT-AES.
    Cooperating Agencies: AR, FL, IL, LA, NY, SC, VT; ARS- Logan, Shafter, Stoneville, Weslaco.
    Reporting: R. Belhe, M. Brownbridge, D. Boucias, S. Jaronski, J. Leland, M. McQuire, D. Steinkraus

    Summary

    Research has progressed on the development of entomopathogenic fungi as microbial control agents for mites, aphids, thrips, glassy-winged sharpshooter, lygus bugs and cerembycids. This research involved a variety of agricultural systems, including cotton, vegetable crops, avocado orchards and forests. A Hirsutella-like organism was the most prevalent pathogen found on dead glassy winged sharpshooter. Beauveria bassiana has been identified as a major pathogen of lygus bugs, and efforts are focusing on development of novel formulations that enhance field stability and efficacy. Neozygites fresenii was shown to be a natural control of cotton aphids for the 11th consecutive year in AR, although efforts to introduce the fungus into Californian cotton fields was not successful. A cultural/biological control strategy for avocado thrips is being investigated; placement of mulch around avocado trees promotes the incidence of insect-pathogenic fungi and other natural enemies. Not-target impact assessment studies are continuing on Bt-silage corn.


    SUBPROJECT 3: Development, evaluation and safety of entomopathogens in cryptic and soil habitats.

    Coordinators: Parwinder Grewal, OH and Edwin Lewis, VA.
    Cooperating Agencies: CA, CT, FL, GA, HI, IL, INHS-IL , LA, MD, NJ, NC, OH, OK, SC, TN, VA; ARS-Wooster, Byron, Sidney, Corvallis, Yakima; USDA FS-CT,MI; Integrated Biocontrol Systems-IN.
    Reporting: M. Barbercheck, D. Bruck, P. Grewal, A. Hajek, S. Jaronski, H. Kaya, L. Lacey, E. Lewis, C. McCoy, D. Shapiro-Ilan, L. Solter, R. Stuart.

    Summary

    Ten state agencies and six USDA field stations submitted research results for this report. Reported research includes many different types of studies that include entomopathogenic nematodes and entomopathogenic fungi. Field or greenhouse trials testing efficacy of insect pathogen-based materials were conducted against nine insect species in seven cropping systems and against plant-parasitic nematodes in two commodities. Twelve species of pathogens were tested for efficacy against five species of agricultural pests in laboratory trials. Several relationships between entomopathogenic nematode infective juvenile longevity and stress tolerance have been discovered in laboratory trials. Studies of the interactions between entomopathogenic nematodes and their symbiotic bacteria with various soil organisms suggest that plant-parasitic nematodes are impacted more than most other soil fauna. One survey of Turkey for entomopathogenic nematodes isolated nematodes from approximately 2% of the samples taken; 4 species were isolated from the area.


    SUBPROJECT 4: Development, evaluation and safety of entomopathogens for veterinary and structural arthropod pests.

    Coordinators: James J. Becnel, David Oi, ARS-Gainesville
    Cooperating Agencies: AL, AR, CA, CT, FL, GA, LA, ME, MS, NC, OK, SC, TN; ARS-Gainesville, Peoria.
    Reporting: T. Andreadis, C. Vossbrinck, J. Fuxa, R. Pereira, J. Becnel, D. Boucias, B. Federici, D. Oi and E. Groden.

    Summary

    Entomopathogenic research on structural pests focused on the control of the red imported fire ant and termites. An integrated approach combining the microsporidium Thelohania solenopsae, parasitic phorid flies, and chemical control showed longer term control of fire ants than chemical treatment alone. Detection protocols, geographic distribution, and biology of T. solenopsae are being established. The effect of other fungal and protozoan pathogens on ants and termites are being evaluated.
    Entomopathogens evaluated against mosquitoes included bacteria, viruses, microsporidia, and a new insect pathogenic algae. A novel recombinant Bacillus thuringiensis subsp. israelensis (Bti) strain and the baculovirus CuniNPV have shown excellent activity against Culex mosquitoes that vector West Nile Virus in the U.S. Basic studies focused on the taxonomy and life histories of new pathogens while other investigations focused on safety, efficacy against the target hosts and improving production and delivery systems of promising agents.

    Impact Statements:
    1. This project impacts biological control and integrated pest management in the Southern Region through the development and integration of entomopathogens into IPM systems. Progress was made in isolating and assessing new bacteria, fungi, microsporidia, nematodes and viruses for control of lepidopteran, homopteran, coleopteran, dipteran, and orthopteran pests of crops (corn, cotton, soybeans, potatoes and other high value crops), forests, rangelands, and man and animals.
    2. Tests for efficacy against insect species, species of plant-parasitic nematodes and one species of slug were conducted throughout the year. Five groups of pathogens were represented in these tests. In addition to efficacy trials, several experiments were conducted to assess environmental tolerance of the pathogens or pathogen-based products.
    3. Collaborative studies by project scientists have led to the discovery of new bio- pesticides for the control of mosquitoes that vector diseases in the United States. This includes a novel recombinant Bacillus thuringiensis subsp. israelensis (Bti) strain and the baculovirus CuniNPV which have shown excellent activity against Culex mosquitoes that vector West Nile Virus in the U.S.
    4. Project scientists also have been able to infect fire ant colonies and document the spread of the pathogen, Thelohania solenopsae. Infected fire ant populations were reduced by 63% and were more susceptible to fire ant bait.
    5. T. solenopsae is now a major component of an areawide IPM program for fire ants. These new technologies can provide safe and effective control and can reduce the amount of chemical pesticides released into the environment.
    Last Modified: unknown

    Date of Annual Report: 04/21/2005

    Report Information:
  • Annual Meeting Dates: 02/27/05 to 02/28/05
  • Period the Report Covers: 01/2004 to 12/2004

    • Participants:
    Brief Summary of Minutes of Annual Meeting:
    MINUTES OF THE 2005 S-301 TECHNICAL COMMITTEE MEETING

    Number and Title of the Regional Project: S-301, Development, Evaluation and Safety of Entomopathogens for Control of Arthropod Pests.

    Location and Date of the Meeting: February 27-28, 2005; Edgefield Inn, Troutdale, Oregon

    Adopted Agenda: See attachment

    Meeting came to order at 8:30 AM, February 27, 2005.

    The meeting called to order by R. P. The first order of business was discussion of the up-coming submission of the renewal for the project. All is on schedule and no problems are anticipated in meeting the deadline of October, 2005 for the final product.

    Denny Bruck, the local organizer gave details of the meeting location and meals to be served. Registration was set at $50.

    SARES Administrative Advisors D. Boethel gave report on project and praised participants for effectiveness in producing reports and proposal renewal. David talked about the new proposed federal budget and the possible effects on Land Grant universities and the research conducted at these institutions. Some discussion followed with participants stating the different levels of impact expected in different universities. Expected impact vary from minimum in states that do not rely heavily on federal funding, to very hard on states such as Maine and others where most of faculty salaries are paid with federal funding.

    Preliminary introductions on the Workshop followed and D. Bruck took command of the meeting to make introductions of Workshop presenters.

    Special minutes: Workshop with Growers, Researchers and Industry representatives. Our goal was to come up with an action that could be taken by the group as a whole. Two presentations were made at the beginning to illustrate two situations where microbials have been successfully implemented:

    Robin Stuart presented a talk on the use of entomopathogenic nematodes in Florida citrus against the citrus root weevil.

    Lerry Lacey presented a talk on using microbials in apple in western Washington.

    Question: How flexible are growers in terms of implementing new methods for pest control?

    a. commodity-specific flexibility

    1. Low-flexibility: potato, row crops, cotton, traditional agricultural crops 2. High-flexibility: tree fruit, greenhouse, nurseries

    Proposed reasons for the differences were education, need for a fast re-entry interval for greenhouse operations and the need for new answers to new problems faced in the nursery business. Also, some crops have pressing pest problems that are not currently being solved by chemical control strategies, so they are looking for other alternatives.

    Grower Needs A suggestion was made that the traditional approach to solving insect problems is to wait until they are true crises. This is not an approach that is amenable with sustainable agriculture. Several reasons were suggested to underlie this situation. A most important one was that funding agencies will only fund research on an existing problem, by which time it is too late to treat is as anything but a crisis.

    Most growers have diversified. In this newly diverse habitat, determining how to integrate various different crops into a cohesive management program is problematic. Suggestion were made by the growers that limited understanding of complexity of the system is a significant impediment management programs. One suggestion was to emphasize a conservation approach to microbial control. This approach is compatible with many of the new practices of sustainable agriculture. The effects of composting and using compost tea were discussed at length. The final consensus, if there was one, was that we dont know enough about what, exactly, comprises compost tea to know what the interactions between this amendment and microbial insecticides might be. This was identified as a potential fruitful direction for further research.

    Stored products were also identified as problematic, especially those that contribute allergens to the environment.

    New treatment thresholds need to be developed. The threshold for microbials might often be lower than those for chemicals. But, these are not available at the time.

    We need to try to come up with recommendations about how to integrate microbials into the whole system.

    Industry needs

    It is a challenge to instruct growers to use products properly. When a product fails due to improper use, it is very costly to the biocontrol industry because the end users tend to think the product is unreliable. The method for getting instructions out to growers that was developed for the chemical industry is not working for biological control, partly because it takes a more sophisticated consumer to use microbials, and partly because a major source of information for the growers is coming directly from the chemical industry.

    We cant expect the majority of growers to make major shifts in management to accommodate microbials.

    Information dissemination is a major problem. The chemical industry has spray manuals, etc. that every county agent has at their disposal. No such guide is available for microbials.

    A proposal was made to develop such a guide. Such a guide should be:

    Regularly updated. Available on the Internet. List relevant information that would enable County Extension personnel to recommend them. Advertised in trade journals. Linked to Pest Management Centers. Organized based on the commodity groups in the SDC 314 proposal.

    The possibility of the group submitting a grant proposal to SARE, or a similar agency, was discussed at length. The deadlines for various regional grant programs will be checked and the group will target one of these programs for a proposal. Possibilities other than SARE: IR-4.

    A committee, comprised of the sub-project chairs, will build a framework for such a web page, and then send out questionnaires to everyone in their Subsections. Ellie Groden volunteered to check into the SARE grants in order to hire someone to coordinate and build the web page. Annual updates will be done in conjunction with the annual meetings.

    Subproject Discussions:

    Subproject 2 Discovery of entomopathogens and their integration and safety in pest management programs for ornamental, vegetable, fruit and nut crops. (Chair, D. Shapiro-Ilan)

    Drion Boucias (Univ of FL) is studying a new Hirsutella spp that is pathogenic to glassy winged sharpshooter.

    Bill Moar (Auburn Univ, AL), is investigating Bt in weevils (sweet potato weevil).

    Lerry Lacey (ARS, Wapato) reported on M. anisopliae on wireworms and virus Vs tuber moth worm.

    Lee Solter (IL) is investigating surfactants combined with S. feltiae for thrips control. Also she is identifying a new microsporidia from black vine weevil in collaboration with Denny Bruck (ARS, Corvallis). Denny Brucks lab is also looking as M. anisopliae for black vine weevil control in ornamentals.

    David Shapiro-Ilan (ARS, Byron) is investigating microbial control of orchard pests. Nematodes are showing promise in control of pecan weevil, plum curculio, and lesser peach tree borer. Fungi are also showing promise for pecan weevil control. Shapiro-Ilan is conducting studies on the genetic basis of beneficial trait deterioration in collaboration with Randy Gaugler (Rutgers, NJ) and Byron Adams (Brigham Young Univ, UT).

    Ed Lewis is conducting studies on entomopathogenic nematode infection dynamics in collaboration with D. Shapiro-Ilan and Jim Campbell (ARS, Manhattan, KS).

    The S-301 Meeting continued on Mon, Feb 28, 2005

    Subproject 4 Discovery of entomopathogens and their integration and safety in pest management programs for medical, veterinary, and structural pests: (Brian Federici leading in place of chairs Jimmy Becnel & David Oi)

    Bob Behle (ARS, Peoria) is working with a new foam formulation for delivery.

    Drion Boucias (Univ FL) reported on Helicosporidium in Culex. Activities include methods of increasing virulence. This pathogen has an extremely wide host range.

    Tom Corell (Earth Bioscience) reported on tick control with Metarhizium anisopliae; testing the F52 strain in field. The primary interest is in homeowner use.

    Roberto Pereira reported on various activities at his location (ARS Gainesville) pertaining to fire ant research. David Oi is looking for new Thelohania in Argentina; and also looking at Vairimorpha field release. A combination of these two pathogens would likely be most efficacious. Also at the Gainesville ARS lab, researchers found a new virus in ants (a picornavirus), which is the first virus reported in ants. A control approach would likely be inoculative or introduction (for virus and above pathogens) resulting in moderate reduction. Pereira is pursuing fungi for fire ant control using a Hirsutella and an unidentified fungus (tentatively Akanthomyces sp.).

    Ellie Groden (Univ ME) is working on an ant being called the European fire ant, which is a continuing significant problem in several localities in Maine. She did survey work in England and found several fungi and a nematode, including putative new species of fungus.

    Brian Federici (Univ CA, Riverside) reported on work on recombinant bacteria for mosquito control; they transferred the toxin from Bacillus sphaericus into Bacillus thuringiensis israelensis (Bti) (both bacteria are already commercial products for mosquitoes). The result of the recombination is greater prod of B. sphaericus toxin per cell. The result is 10-20x efficacy compared to either wild type. Efficacy mostly vs. Culex but also may have potential Vs malaria carrying mosquitoes.

    Subproject 1 Discovery of entomopathogens and their integration and safety in pest management programs for major acreage crops (Bob Behle, Chair).

    Sugar beets: Mark Boetel (North Dakota State Univ) is working on fungi for control of the sugar beet maggot using Beauveria and Metarhizium. He is collaborating with Stefan Jaronski (ARS, Sidney) and conducting lab and field trials. Jaronski (ARS, Sidney) working with Metarhizium anisopliae strain F52 for sugar beet maggot control, observed reduced efficacy with decreasing soil moisture. Metarhizium seems to work well when the insect density is not too high; at higher densities the pathogen approach will probably have to be integrated with other strategies. Jaronski is also looking at wireworms and Collembola control with fungi.

    Cotton: Bill Moar (Auburn University, AL) is working with Bt cotton. One project focuses on resistance management; the team is developing a standard protocol to detect resistance. Also Moar is working on developing resistance to H. zea in lab.

    Jarrod Leland (ARS, Stoneville) is researching microbial control of cotton pests. He is collaborating with Mickey McGuire (ARS, Shafter) investigating new isolates of B. bassiana for control of Lygus. Leland is also looking at Neozygites in cotton aphid suppression (and for consideration in soybean aphid control).

    Mickey McGuire (ARS, Shafter) reported on their focus on Lygus control with microbials in CA cotton. They observed high levels of control with B. bassiana fungi in alfalfa (they used alfalfa because Lygus populations are more consistent than in cotton). They have developed molecular markers for the B. bassiana strains so that they can follow infections/persistence of these strains following field application.

    Alfalfa: Rosalind James (ARS, Logan) investigated non-target effects of hyphomycete use in alfalfa on pollinators. Also they have developed PCR markers to identify chalkbrood infections. Additionally, in collaboration with J. Leland, she is working on Varroa mite control with Metarhizium anisopliae.

    Potatoes: E. Groden reported on studies using B. bassiana for Colorado potato beetle control, looking at interaction between introduced and endemic fungal strains.

    L. Lacey (ARS, Wapato), is investigating potato tuber moth control, a significant emerging pest. They are looking at granulosis virus as well as fungi and nematodes for control. Also they are studying control of wireworms in potato; nematodes do not appear to be promising. Metarhizium may be an option.

    Other: Drion Boucias (U FL) is working on Thripinema in collaboration with J. Funderburk (U FL)for thrips control.

    Subproject 3 Discovery of entomopathogens and their integration and safety in pest management programs for urban and natural landscapes (Roberto Pereira led the meeting in place of co-chairs Parwinder Grewal and Ed Lewis).

    Lee Solter (IL) looking at host specificity microsporidia, and field trials for gypsy moth control; collaboration is with V. DAmico (USFS, Newark).

    Tom Corell (Earth BioScience) is cooperating with Ann Hayek (Cornell, NY) on M. anisopliae control of Asian longhorn beetle using bands of the fungus around the tree. Also Corell is working on white grub control in turf.

    DeAnna Borchardt (U of DE) working with Vince DAmico (USFS, Newark) is using a voltage clamp to study Bt toxin effects in gypsy moth.

    Stefan Jaronski reported on grasshopper control with fungal agents on rangelands. He is investigating an attracticide and strip-treatment approach using GHA strain, but temperature tolerance is problematic due to behavioral fever. Also he is looking at Mormon cricket control with fungal agents. Jaronski is collaborating with Don Roberts (UT St. Univ.), they are looking for new pathogens in Mormon cricket.

    FINAL BUSINESS MEETING Information - see attachment


    URL: Copy of minutes

    Accomplishments:
    CROSS-SUBPROJECT EFFORTS:

    A catalogue of classical biological control introductions of entomopathogens against a great diversity of insects and mites has been prepared and will be published in a Forest Service publication and on the website. The pathogenicity of three strains of Beauveria bassiana towards adults of the alfalfa leafcutting bee could only be demonstrated at very high concentrations; therefore this fungus can be safely used in fields where these bees are released. (USDA-ARS, Logan, UT). A molecular marker method based on PCR has been developed for identifying and detecting infections of Ascosphaera spp. (chalkbrood) in the alfalfa leafcutting bee (USDA-ARS, Logan, UT).

    SUBPROJECT 1. Development, evaluation and safety of entomopathogens for leaf feeding insect defoliators.

    Coordinator: Robert Behle, USDA, ARS-Peoria

    Summary

    Entomophaga maimaiga (NY) and several microsporidia isolated from European Lymantria dispar (L.) populations (INHS-IL; USDA FS-CT; CT Ag. Expt. Stn.; Bulg. Academy of Sciences; Fach. Eberswalde, Germany; BOKU, Vienna; Illinois DNR and DOA) continue to be evaluated for gypsy moth control. Field studies evaluated non-target effects and horizontal transmission of microsporidia. Release of three species of gypsy moth microsporidia has been proposed for Illinois. Nosema from the Eastern tent caterpillar, Malacosoma americanum (USDA-ARS Stoneville; Univ. of KY; INHS; Iowa State Univ.) and a microsporidium from the black vine weevil, Otiorynchus sulcatus (USDA-ARS, Corvallis; INHS) are also being evaluated for use as biological control agents. Season-long treatments of codling moth granulovirus (CpGV) resulted in significantly fewer deep entries and surviving larvae, but did not reduce the proportion of fruit damaged by codling moth despite pest population reduction. Viral formulations are being evaluated in the field and under storage conditions. The Cyd-X and Virosoft formulations were stable at 95ÚF for several months; Carpovirusine formulation only had good survival at the lower temperatures (USDA-ARS, Yakima). Lignin-based spray-dried formulation AgMNPV provided protection of the virus from simulated sunlight degradation in the lab (USDA-ARS, Peoria) but not in the Brazilian fields (EMBRAPA-Londrina). Studies on Beauveria bassiana population genetics indicated the displacement of naturally occurring populations by the commercial strains and some degree of genetic transfer from the commercial strains into the indigenous population (University of Maine; USDA-ARS, Ithaca, NY). B. bassiana and Metarhizium anisopliae conidia did not infect Collembola, although significantly fewer eggs were produced probably due to poor diet quality. A Bt diet and fungal metabolites also did not affect these insects (Univ. of Vermont, Burlington, VT).

    SUBPROJECT 2: Development, evaluation and safety of entomopathogens for homopteran and other piercing-sucking insects.

    Coordinator: Michael Brownbridge, Univ. of VT-AES, & David Shapiro-Ilan, USDA-ARS, Byron, GA.

    Summary

    Thripinema fuscum affected the fecundity of Frankliniella fusca, indicating the potential of the nematode to suppress F. fusca populations in peanut (Univ. of FL). Steinernema feltiae and Heterorhabditis indica kill the Western flower thrips, Frankliniella occidentalis Pergande, in glasshouse Gerbera daisies but access to the flowerhead by sprayed nematodes is limited by flower morphology. (University of Illinois; INHS, IL). Entomopathogenic fungi and nematodes may be added to mulches to bioenhance and improve their suppressive effect on avocado thrips. (Univ. of Vermont, Burlington, VT; UC Riverside, CA). Fungal pathogens, including Pandora neoaphidis, were identified infecting the invasive soybean aphid, recently introduced from China. The fungal pathogen Neozygites tanajoae of cassava green mite has been described, methods for culture and storage have been developed and pathogenicity and host specificity have been documented. (Cornell Univ., NY). Metarhizium anisopliae var. acridum isolate CG423 was demonstrated to be pathogenic to the false spider mite Brevipalpus phoenicis Geijskes with 90% mortality by the 8th day.(Univ. of FL). A survey of the glassy-winged sharpshooter Homalodisca coagulata demonstrated that mycosed GWSS occurred at more than 25% of the sampling sites with more than 75% of the mummified insects harboring Hirsutella. (Univ. of FL). A microsporidium found in Pseudoscymnus tsugae, a predator of the woolly hemlock adelgid, may only have serious deleterious effects at high prevalences in the host colonies. (NJ Dept. of Agric., INHS).

    SUBPROJECT 3: Development, evaluation and safety of entomopathogens in cryptic and soil habitats.

    Coordinators: Parwinder Grewal, OH and Edwin Lewis, CA.

    Summary

    Metarhizium anisopliae (strain F52, Earth BioSciences, Fairfield, CT) was tested against Asian longhorned beetle (Cornell Univ., NY) and the black vine weevil, Otiorhynchus sulcatus (USDA-ARS, Corvallis, OR). Beauveria bassiana and Paecilomyces tenuipes, Steinernema oregonense, and Bacillus thuringiensis were also bioassayed against the black vine weevil (USDA-ARS, Corvallis, OR). M. anisopliae also showed potential for control of adult western cherry fruit fly emerging through fungus treated soil (USDA-ARS, Yakima). Entomopathogenic nematodes were shown to reduce plum curculio beneath fruit trees (Utah State University; USDA-ARS, Wapato WA), and overwintering stages of Oriental fruit moth OFM (Washington State University; USDA-ARS, Wapato WA). Codling moth larvae were nearly totally controlled in mulched plots that were treated with 1 billion nematodes per acre (USDA-ARS, Wapato WA). Steinernema carpocapsae caused over 90% suppression of navel organgeworm at a rate of 0.4 billion nematodes per acre of pistachios (USDA-ARS, Parlier CA & Wapato WA). Genetic studies on trait deterioration have been conducted with entomopathogenic nematodes (USDA-ARS Byron GA, NJ, USDA-ARS Newark DE, Brigham Young Univ.). Mixed strains of the Steinernema riobrave from Texas were tested against citrus root weevil larvae (University of Florida, IFAS, CREC; USDA-ARS Byron GA).

    SUBPROJECT 4: Development, evaluation and safety of entomopathogens for veterinary and structural arthropod pests

    Coordinators: James J. Becnel, David Oi, ARS-Gainesville

    Summary

    Entomopathogenic research on structural pests focused on the control of the imported fire ants, (Solenopsis invicta, S. richteri, and their hybrid) and the invasive European fire ant, Myrmica rubra. A natural infection of the microsporidian, Thelohania solenopsae was reported for the first time from the single-queen form of the red imported fire ant, as well as an infection in the hybrid imported fire ant. Laboratory transmission of another microsporidium, Vairimorpha invictae was accomplished for the first time and there were significant reductions in the growth of infected S. invicta colonies. A new virus was sequenced and characterized from S. invicta, and new fungal pathogens were isolated from this ant species. Isolates of Metarhizium anisopliae and Beauveria bassiana obtained from M. rubra were more pathogenic than a commercial B. bassiana strain. Compensatory mortality was observed with simultaneous exposure of M. rubra to fungi and boric acid ant baits. Entomopathogens evaluated against veterinary pest included bacteria, viruses, fungi, microsporidia, and a new insect pathogenic algae. New mosquitocidal recombinant strains of Bacillus thuringiensis subsp. morrisoni (PG-14) and B. thuringiensis subsp. jegathesan (Btj) were developed with improved efficacy against Culex and Aedes spp. The infectivity and pathogenicity of the baculovirus, OscoNPV were evaluated in laboratory bioassays against ten species of mosquitoes native to the northeastern US. The host range was restricted to Aedes and Ochlerotatus spp. A cytoplasmic polyhedrosis virus was isolated from the mosquito Uranotaenia sapphirina (UsCPV) and molecular data revealed a distant relationship with the genus Cypovirus. Various species of microsporidia have been sequenced to study relationships among the Microsporidia. Basic studies on Helicosporidium sp., (an achlorophytic green algae) have demonstrated that it infects various lepidopteran, dipteran, and coleopteran hosts. Beauveria bassiana was isolated in Minnesota from female blacklegged ticks, Ixodes scapularis. The nuclear small subunit ribosomal RNA internal transcribed spacer regions 1 and 2 of these isolates was sequenced and compared with those from other strains and species of Beauveria.

    Impact Statements:
    1. This project impacts biological control and integrated pest management in the Southern Region through the development and integration of entomopathogens into IPM systems. Progress was made in isolating and assessing new bacteria, fungi, microsporidia, nematodes and viruses for control of insects (Lepidoptera, Homoptera, Coleoptera, Diptera, Hymenoptera and Thysanoptera) as well as ticks and mites. Target pests include those attacking crops (corn, cotton, soybeans, potatoes, fruits, ornamentals.
    2. Tests for efficacy against new invasive species, which represent very serious threats to the US economy, provide information on new efficient methods of controlling these pests. Control methods vary from use of entomopathogens permanently established in the environment to biopesticidal applications of formulations with enhanced desirable characteristics. Target pests vary from general pests, such as the red imported fire ant, to more specific ones such as the soybean aphid and the gypsy moth
    3. The development of new entomopathogens as agents for insect pest control impacts both traditional and organic agriculture. New methods of application of entomopathogens to soil and/or mulch can provide efficient control of pests in diverse production systems.
    4. New recombinant strains of Bacillus thuringiensis with improved efficacy against Culex and Aedes spp., and new viruses offer alternative methods for control of diseases-carrying mosquitoes. These entomopathogens can help prevent epidemics of diseases in humans and in domestic animals.
    5. A catalogue of classical biological control introductions of entomopathogens will facilitate future research on microbial control of both new invasive and native insect pests.
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