Progress 12/04/98 to 12/03/03
Outputs
1. What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter? Commercial interest in using living microbial agents for controlling agricultural pests such as weeds, insects and plant pathogens is driven by an increasing public demand for environmentally-friendly, sustainable agricultural practices. The widespread use of chemical pesticides is often viewed as non-sustainable agricultural practices. An integrated pest management (IPM) approach using biological control agents (biopesticides) in conjunction with chemical pesticides is recognized as a sustainable pest management system, which reduces chemical pesticide usage and deters the development of pest resistance to the chemical control. A significant constraint to the commercial use of living microbial agents in agriculture is the lack of low-cost methods for producing and formulating stable and effective
microbial products. While living microbial agents can be produced using solid-substrate or liquid culture methods, liquid culture fermentation processes are generally the most cost-effective. Optimized deep-tank, liquid fermentation processes for the production of living microbial agents must take into account not only product yield, but also microbial stability and product efficacy. Likewise, commercially viable microbial products require formulations that provide stabilized, effective biopesticides in a form that is conducive to application technology commonly used in agriculture. Our three major research goals will be to (1) develop low-cost, deep-tank fermentation methods for the production of stable, effective living microbial propagules for use as biocontrol agents, (2) develop methods for drying and formulating fungal and bacterial microbial biopesticides that enhance the viability and efficacy of these agents during storage and application, respectively, (3) develop bait
and kill formulations for use with existing chemical and biological pesticides to decrease the amount of toxicant required for effective control. Public concerns over the widespread use of chemical pesticides and increased pest resistance to safe chemical controls have heightened interest in developing environmentally friendly pest control measures. The use of microbial pathogens as biopesticides for controlling weeds, insects, and plant diseases is an acceptable alternative control measure. Need for microbial products that control plant pests remains high. While plant and insect pathologists have identified hundreds of microbial agents over the past 20 years that show good potential for use as biopesticides, few living microbes have been commercially developed as biopesticides. In large part, the limited success in commercializing microbial agents is due to a lack of economical production and formulation processes that deliver microbial products with a reasonable shelf-life and that
provide consistent pest control under field conditions. These technical constraints to the use of biopesticides require research designed to understand how the nutritional and environmental conditions present during the production, formulation, and application processes affect biopesticide yield, stability and biocontrol efficacy. Unless these problems are addressed, the widespread use of biopesticides will not become a reality. This research focuses on the development of cost effective fermentation and formulation processes for microbial biopesticides. These efforts contribute to National Program 304 "Crop Protection and Quarantine"; Component 5: Pest Control Technologies, and Component 9: Biological Control of Weeds (70%). The expected outcome from this research is the commercial use of microbial biopesticides for insect and weed control, which will lead to additional markets for the agricultural commodities used as ingredients in the fermentation and formulation processes,
thus contributing to National Program 306 "Quality and Utilization of Agricultural Products;" Component 2: New Processes, New Uses, and Value- Added Foods and Biobased Products (30%). 2. List the milestones (indicators of progress) from your Project Plan. This research project, 3620-22410-004-00D, terminated on 12/03/03 and was bridged to project 3620-22410-008-00D. A prospectus has been accepted and a new project plan is currently being written for inclusion in the NP 304 Crop Protection and Quarantine Panel Review scheduled for late 2004 or early 2005. The new research project is being developed to continue current research. Provided below are objectives for this project. Objective 1: Develop low-cost, deep-tank fermentation methods for the production of stable, effective living microbial propagules for use as biocontrol agents or probiotic feed additives. - Optimize production methodology for the potential fungal bioherbicide Colletotrichum truncatum. - Develop production
technology for spores of the fungal bioinsecticide Paecilomyces fumosoroseus. - Evaluate promising microbial biocontrol agents for suitability to production using deep-tank fermentation and develop production technology. Objective 2: Develop methods for drying and formulating fungal and bacterial microbial biopesticides which enhance the viability and stability of these agents during storage. - Develop stabilization technology for liquid culture produced spores of the bioinsecticide Paecilomyces fumosoroseus. - Identify nutritional conditions during cell growth and propagule formation which enable the microbial propagule to survive the drying process. - Develop air- and freeze-drying techniques which abate the deleterious events associated with dehydration. - Define storage conditions (temperature, humidity, gaseous environment) and rehydration protocols for dried biopesticidal propagules which maximize cell survival. Objective 3: Identify potential bacterial or fungal biocontrol
agents for the control of Fusarium graminearum, the causative agent of wheat head scab (HSW). - Through selective screening protocols, identify bacterial and fungal antagonists of F. graminearum. - Determine dose responses for HSW antagonists in greenhouse and field trials. - Evaluate wheat cultivar influence on HSW antagonist biocontrol efficacy. - Evaluation of HSW biocontrol efficacy of additional microbial strains obtained from wheat anthers. - Efficacy testing of antagonist mixtures against HSW: greenhouse and field trials. - Determination of liquid culture growth kinetics and efficacy of top HSW antagonists. - Optimization of liquid culture production for superior HSW antagonist. - Field testing of liquid culture produced cell pastes and freeze-dried preparations of selected HSW antagonist. 3. Milestones: A. List the milestones (from the list in Question #2) that were scheduled to be addressed in FY 2004. How many milestones did you fully or substantially meet in FY 2004 and
indicate which ones were not fully or substantially met, briefly explain why not, and your plans to do so. All milestones scheduled for FY 2004 for Objectives 1 and 2 were met. Milestones for Objective 3 are now included in the research conducted in project 3620-22410-007-00D, "Microbial production and formulation technologies for biocontrol of fungal plant diseases." - Develop production technology for spores of the fungal bioinsecticide Paecilomyces fumosoroseus. - Evaluate promising microbial biocontrol agents for suitability to production using deep-tank fermentation and develop production technology. - Define storage conditions (temperature, humidity, gaseous environment) and rehydration protocols for dried biopesticidal propagules which maximize cell survival. B. List Milestones that you expect to address over the next 3 years (FY 2005, 2006, 2007). What do you expect to accomplish, year by year, over the next 3 years under each milestone? This research project,
3620-22410-004-00D, terminated on 12/03/03 and was bridged to project 3620-22410-008-00D. A prospectus has been accepted and a new project plan is currently being written for inclusion in the NP 304 Crop Protection and Quarantine Panel Review scheduled for late 2004 or early 2005. Proposed milestones for the new project plan are listed in the Report of Progress (AD-421) for bridging project 3620- 22410-008-00D. 4. What were the most significant accomplishments this past year? A. Single most significant accomplishment during FY 2004: This research project, 3620-22410-004-00D, terminated on 12/03/03 and was bridged to project 3620-22410-008-00D. A prospectus has been accepted and a new project plan is currently being written for inclusion in the NP 304 Crop Protection and Quarantine Panel Review scheduled for late 2004 or early 2005. Accomplishments for FY 2004 are listed in the Report of Progress (AD-421) for 3620-22410-008-00D. B. Other significant accomplishment(s), if any:
None. C. Significant activities that support special target populations: None. D. Progress Report: None. 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. This research project, 3620-22410-004-00D, terminated on 12/03/03 and was bridged to project 3620-22410-008-00D. A prospectus has been accepted and a new project plan is currently being written for inclusion in the NP 304 Crop Protection and Quarantine Panel Review scheduled for late 2004 or early 2005. Major accomplishments of the current project built on a previous project 3620-41000-057-00D. Previous studies led to the development of deep-tank fermentation methods for the production of three promising microbial biopesticides: Colletotrichum truncatum, a fungal biopesticide for control of the weed hemp sesbania; Xanthomonas campestris, a bacterial biopesticide used for controlling the weedy grass; Poa annua (annual bluegrass); Paecilomyces fumosoroseus, a fungal
pathogen of numerous insect pests including the silverleaf whitefly; and Mycoleptodiscus terrestris, a fungal biocontrol agent of the invasive aquatic weed Hydrilla verticillata (hydrilla). In these studies, the nutritional composition of liquid culture media was optimized to maximize yields, propagule stability and biocontrol efficacy. This novel research showed that the diets fed microbes during the production process could have a profound influence on the quantity and quality of the biopesticidal product. A major discovery derived from CRIS #3620-41000-057-00D was the development and patenting of a method for producing infective, stable spores of the bioinsecticide Paecilomyces fumosoroseus (U.S. Patent #5, 968,808, October 19, 1999). Work with the production and stabilization of P. fumosoroseus spores continues to be a major focus of the present CRIS project. It is expected that our P. fumosoroseus spore production process will become a commercial method for producing this
promising bioinsecticide. A novel approach to producing biopesticidal spores of P. fumosoroseus is on-site production in portable fermentation equipment. In order to use portable fermentation equipment for the on-site production and delivery of spores of P. fumosoroseus, process conditions were developed which favor the rapid production of P. fumosoroseus spores while inhibiting the growth of undesirable microbes. High concentrations of P. fumosoroseus spores were rapidly produced in a portable fermentor with little or no bacterial contamination when the pH of the production medium was lowered and appropriate nutrient concentrations were provided in the production medium. These studies have demonstrated that the on- site production of fungal biocontrol agents is feasible and, specifically, that high concentrations of P. fumosoroseus can be rapidly produced using an aseptic, portable fermentation system. Other insect targets for the bioinsecticide P. fumosoroseus have been
identified including the Formosan subterranean termite and the Mexican bean beetle. Nutritional and environmental conditions were identified that support the production of a stable form of the fungal biocontrol agent Mycoleptodiscus terrestris (MT), a promising biocontrol agent for the invasive aquatic weed hydrilla (Hydrilla verticillata). These MT preparations were dried with excellent stability and demonstrated superior biocontrol efficacy against hydrilla. This production and stabilization process has been patented jointly by ARS and the U.S. Army Corps of Engineers (Mycoherbicidal compositions and methods of preparing and using the same, U.S. Patent #6,569,807). The optimization of this process supported the large-scale production of stable, effective MT preparations that were field-tested against hydrilla in Florida and Texas by scientist from the U.S. Army Corps of Engineers, the University of Florida, and an industrial partner. The development of a non-chemical control
measure for hydrilla, a serious aquatic weed in much of the Southern U.S., will be a welcome tool in the fight to control this menacing invasive weed species. Effective 10/01/00, project 3620-41000-070-00D entitled "Formulation and adjuvants for biological controls and insect-active volatiles" was combined with project 3620-22410-004-00D. Over the life of this incorporated project, several formulations and adjuvants have been developed and tested for use with microbial pesticides and for use in reducing the amounts of chemical pesticides needed for control. Corn flour, gluten, and lignin were all tested as adjuvants for reducing wash- off by rain and for preventing UV light from inactivating microbial pesticides. Three patents have been issued concerning these formulations. Prior to 2000, adjuvants were tested with a product called Slam Insecticide, which is being used to control adult corn rootworms with a very low amount of insecticide. Slam must remain on the plant surface for
several weeks after application to be economical. Rainfall quickly washes slam from the plant. As a result of our work, slam was reformulated to include gluten to resist wash-off by rainfall, thus extending the residual activity of the application. Following this work, cooperation with an industry partner led to the development and commercialization of a spray adjuvant to improve control of corn rootworms with one-tenth the normal amount of insecticide. This adjuvant, Cidetrak CRW, is currently available to growers. Another technology developed through this project, was the Fruitsphere for control of fruit flies in orchards. In a cooperative project with university scientists, a fruit mimicking spheres composed of corn flour, corn syrup, and sugar with a coating in insecticide continues to progress toward commercialization. Current efforts are directed at using biodegradable starch-based plastics to mold the fruit mimic. Recent formulation and stabilization research has led to
the development of a spray-dried formulation process for imparting solar stability to spores of the bioinsecticidal fungus B. bassiana for improved persistence under field conditions. 6. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? Technology transfer activities for this project, which terminated on 12/3/03, are covered in the Report of Progress (AD-421) for bridging project 3620-22410-008-00D. 7. List your most important publications in the popular press and presentations to organizations and articles written about your work. Popular and scientific publications for this project, which terminated on 12/3/03, are covered in the Report of Progress (AD-421) for bridging project 3620-22410-008-00D.
Impacts
(N/A)
Publications
- WRIGHT, M.S., JACKSON, M.A., CONNICK, W. J. POTENTIAL USE OF PAECILOMYCES FUMOSOROSEUS FOR CONTROL OF THE FORMOSAN SUBTERRANEAN TERMITE COPTOTERMES FORMOSANUS SHIRAKI. SOCIETY FOR INVERTEBRATE PATHOLOGY. 2003. v. 1: ABSTRACT p. 104.
- JACKSON, M.A. CULTURE MEDIA DESIGN FOR FERMENTATION PROCESSES. BIOTECHNOLOGY PROCESSES WORKSHOP, UNIVERSIDAD AUTONOMAS DE NUEVO LEON, MONTERREY, MEXICO. 2002. v. 1: ABSTRACT p. 34.
- KARGALIOGLU, Y., JACKSON, M.A. LIQUID CULTURE FERMENTATION PRODUCTION OF MICROSCLEROTIA OF MYCOLEPTODISCUS TERRESTRIS AS A BIOHERBICIDE FOR CONTROL OF HYDRILLA VERTICILLATA. AMERICAN SOCIETY FOR MICROBIOLOGY. 2003. v. 1: ABSTRACT p. 470.
- BEHLE, R. W. FORMULATIONS OF BIOPESTICIDES. BIOTECHNOLOGY PROCESSES WORKSHOP, UNIVERSIDAD AUTONOMAS DE NUEVO LEON, MONTERREY, MEXICO. 2002. v. 1: ABSTRACT p. 40.
- BEHLE, R.W. FORMULATIONS TO IMPROVE BACULOVIRUS INSECTICIDES. PROCEEDINGS OF SICONBIOL. 2003. v. 1: ABSTRACT p. 43.
- LELAND, J.E., BEHLE, R.W. DEVELOPMENT OF MYCOINSECTICIDES FOR CONTROLLING TARNISHED PLANT BUGS ON WILD HOST PLANTS. NATIONAL COTTON COUNCIL BELTWIDE COTTON CONFERENCE. 2003. v. 1: PROCEEDINGS P. 954-965.
- JACKSON, M.A., CLIQUET, SL., ITEN, L.B. MEDIA AND FERMENTATION PROCESSES FOR THE RAPID PRODUCTION OF HIGH CONCENTRATIONS OF STABLE BLASTOSPORES OF THE BIOINSECTICIDAL FUNGUS PAECILOMYCES FUMOSOROSEUS. BIOLOGICAL SCIENCE AND TECHNOLOGY. 2003. v. 13, p. 23-33.
- SHEARER, J.F., JACKSON, M.A. MYCOHERBICIDAL COMPOSITIONS AND METHODS OF PREPARING AND USING THE SAME. 2003. U.S. PATENT 6,569,807.
- SHAPIRO, D.I., BEHLE, R.W., MCGUIRE, M.R., LEWIS, E.E. FORMULATED ARTHROPOD CADAVERS FOR PEST SUPPRESSION. 2003. U.S. PATENT 6,524,601.
- VEGA, F.E., JACKSON, M.A., MERCADIER, G., POPRAWSKI, T.J. THE IMPACT OF NUTRITION ON SPORE YIELDS FOR VARIOUS ENTOMOPATHOGENS IN LIQUID CULTURE. WORLD JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY. 2003. v. 19. p. 363-368.
- WRIGHT, M.S., JACKSON, M.A., CONNICK, W.M. USE OF PAECILOMYCES SPP. AS PATHOGENIC AGENTS FOR CONTROL OF TERMITES. AMERICAN SOCIETY FOR MICROBIOLOGY. 2003. v. 1: ABSTRACT p. 439.
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Progress 10/01/02 to 09/30/03
Outputs
1. What major problem or issue is being resolved and how are you resolving it? Commercial interest in using living microbial agents for controlling agricultural pests such as weeds, insects, and plant pathogens or as probiotic feed additives for livestock is driven by an increasing public demand for environmentally friendly, sustainable agricultural practices. The widespread use of chemical pesticides and antibiotic feed additives are often viewed as non-sustainable agricultural practices. An integrated pest management (IPM) approach using biological control agents (biopesticides) in conjunction with chemical pesticides is recognized as a sustainable pest management system, which reduces chemical pesticide usage and deters the development of pest resistance to the chemical control. Likewise, the use of living microorganisms as feed additives (probiotics) has been shown to improve animal health, weight gain, and feed conversion efficiency. Effective probiotic feed
additives can reduce or eliminate the use of antibiotics in feeds. A significant constraint to the commercial use of living microbial agents in agriculture is the lack of low-cost methods for producing and formulating stable and effective microbial products. While living microbial agents can be produced using solid-substrate or liquid culture methods, liquid culture fermentation processes are generally the most cost-effective. Optimized deep-tank, liquid fermentation processes for the production of living microbial agents must take into account not only product yield, but also microbial stability and product efficacy. Likewise, commercially viable microbial products require formulations that provide stabilized, effective biopesticides in a form that is conducive to application technology commonly used in agriculture. Our three major research goals will be to (1) develop low-cost, deep-tank fermentation methods for the production of stable, effective living microbial propagules for
use as biocontrol agents or probiotic feed additives; (2) develop methods for drying and formulating fungal and bacterial microbial biopesticides that enhance the viability and efficacy of these agents during storage and application, respectively; (3) develop bait and kill formulations for use with existing chemical and biological pesticides to decrease the amount of toxicant required for effective control. 2. How serious is the problem? Why does it matter? Public concerns over the widespread use of chemical pesticides and increased pest resistance to safe chemical controls have heightened interest in developing environmentally friendly pest control measures. The use of microbial pathogens as biopesticides for controlling weeds, insects, and plant diseases is an acceptable alternative control measure. Need for microbial products that control plant pests remains high. While plant and insect pathologists have identified hundreds of microbial agents over the past 20 years that show good
potential for use as biopesticides, few living microbes have been commercially developed as biopesticides. In large part, the limited success in commercializing microbial agents is due to a lack of economical production and formulation processes that deliver microbial products with a reasonable shelf-life and that provide consistent pest control under field conditions. These technical constraints to the use of biopesticides require research designed to understand how the nutritional and environmental conditions present during the production, formulation, and application processes affect biopesticide yield, stability and biocontrol efficacy. Unless these problems are addressed, the widespread use of biopesticides will not become a reality. Likewise, the use of chemical antibiotic feed additives has been implicated in hastening the development of antibiotic-resistant bacteria, thus mitigating the usefulness of various classes of antibiotics for human and animal health. Many
countries in Europe have already banned the prophylactic use of chemical antibiotics in animal feeds. In many cases, living microbial feed additives have been shown to provide health and weight gain benefits comparable to those seen with chemical antibiotic supplementation. Since the consistent efficacy of these products depends on the delivery of a living microbe as a feed additive, many of the technical constraints applicable to biopesticides also limit the commercialization of probiotics. The widespread use of probiotic feed additives will require the development of low-cost production methods that yield stable, effective microbial feed additives. 3. How does it relate to the National Program(s) and National Program Component(s) to which it has been assigned? National Program 304, Crop Protection Quarantine (70%) and National Program 306, Quality and Utilization of Agricultural Products (30%) This research focuses on the development of cost effective fermentation and
formulation processes for mass-producing microbial biopesticides that contributes to National Program 304, Crop Protection Quarantine (70%). The expected outcome from this research is the commercial use of microbial biopesticides and probiotic feed additives, which will lead to additional markets for the agricultural commodities used as nutrients in the fermentation and formulation processes, thus contributing to National Program 306, Quality and Utilization of Agricultural Products (30%). In addition, the use of biopesticides will reduce pest damage to plant products thereby improving product quality and marketability. 4. What were the most significant accomplishments this past year? A. Single Most Significant Accomplishment during FY 2003: In order to conduct field trials on the use of Mycoleptodiscus terrestris (MT) for control of the invasive aquatic weed, hydrilla, fermentation processes that led to the production and stabilization of superior preparations of MT were needed.
Nutritional and environmental conditions were identified during deep-tank fermentation studies at USDA-ARS National Center for Agricultural Utilization Research (NCAUR), Peoria, IL, Crop Bioprotection Research Unit, that led to the production of MT preparations with superior biocontrol efficacy against hydrilla. The optimization of this process supported the large-scale production of stable, effective MT preparations that were field-tested against hydrilla in Florida and Texas by scientist from the U.S. Army Corps of Engineers, the University of Florida, and an industrial partner. The development of a non-chemical control measure for hydrilla, a serious aquatic weed in much of the Southern U.S., will be a welcome tool in the fight to control this menacing invasive weed species. B. Other Significant Accomplishment(s), if any: Nematodes are an effective insect control agent that is difficult to produce and formulate using typical chemical production techniques. In order to develop
formulations that enhance nematode survival and efficacy, collaborative research studies were conducted between ARS scientists at NCAUR and Byron, GA. As a result of this research, a formulation technique was developed that coats nematode-infected cadavers and benefits handling and survival of these biocontrol agents (Shapiro et al., US Patent #6,524,601 B1). This technology enhances the commercial potential of nematodes as a natural, non-chemical insect control agent and licensing of this technology is being pursued by an industrial partner. C. Significant Activities that Support Special Target Populations: None. D. Progress Report: Over the past year, nutritional and environmental conditions were optimized for the production and stabilization of the fungal bioherbicide, Mycoleptodiscus terrestris, a fungal pathogen and potential biocontrol agent for the aquatic weed hydrilla. Results from these studies yielded a method for producing high concentrations of a stable, infective
form of the fungus. In collaboration with an industrial partner, this fungal bioherbicide is being field tested in Texas and Florida for hydrilla control. Continued collaborative studies with ARS scientists at the USDA- ARS Southern Regional Research Center, New Orleans, LA, on subterranean termites have demonstrated the excellent potential of this biocontrol agent as a biologically based termiticide. In addition, research continues on the development of formulations for the insect biocontrol fungus, Beauveria bassiana, for insect control in cotton and on the use of caffeine as a slug and snail control agent for greenhouses. 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. While this research project is in its fourth year, it builds on our previous research project, 3620-41000-057-00D, Production and Stabilization of Microbial Bioherbicides. This previous research led to the development of deep-tank fermentation
methods for the production of three promising microbial biopesticides; Colletotrichum truncatum, a fungal biopesticide for control of the weed hemp sesbania, Xanthomonas campestris, a bacterial biopesticide used for controlling the weedy grass, Poa annua (annual bluegrass) and Paecilomyces fumosoroseus, a fungal pathogen of numerous insect pests including the silverleaf whitefly. In these studies, the nutritional composition of liquid culture production media were optimized to maximize yields, propagule stability and biocontrol efficacy. This novel research showed that the diets fed microbes during the production process could have a profound influence on the quantity and quality of the biopesticidal product. A major discovery derived from research project 3620-41000-057-00D was the development and patenting of a method for producing infective, stable spores of the bioinsecticide P. fumosoroseus (U.S. Patent #5,968,808, October 19, 1999). Work with the production and stabilization
of P. fumosoroseus spores continues to be a major focus of the present research project. It is expected that our P. fumosoroseus spore production process will become a commercial method for producing this promising bioinsecticide. A novel approach to producing biopesticidal spores of P. fumosoroseus is on-site production in portable fermentation equipment. In order to use portable fermentation equipment for the on-site production and delivery of spores of P. fumosoroseus, process conditions were developed which favor the rapid production of P. fumosoroseus spores while inhibiting the growth of undesirable microbes. High concentrations of P. fumosoroseus spores were rapidly produced in a portable fermentor with little or no bacterial contamination when the pH of the production medium was lowered and appropriate nutrient concentrations were provided in the production medium. These studies have demonstrated that the on-site production of fungal biocontrol agents is feasible and,
specifically, that high concentrations of P. fumosoroseus can be rapidly produced using an aseptic, portable fermentation system. Other insect targets for the bioinsecticide P. fumosoroseus have been identified including the Formosan subterranean termite and the Mexican bean beetle. Research project 3620-41000-070-00D, Formulation and Adjuvants for Biological Controls and Insect Attractants, was combined with the current research project in 2000. Over the life of this incorporated project, several formulations and adjuvants have been developed and tested for use with microbial pesticides and for use in reducing the amounts of chemical pesticides needed for control. Corn flour, gluten, and lignin were all tested as adjuvants for reducing wash-off by rain and for preventing UV light from inactivating microbial pesticides. Three patents have been issued concerning these formulations. Prior to 2000, adjuvants were tested with a product called Slam Insecticide, which is being used to
control adult corn rootworms with a very low amount of insecticide. Slam Insecticide must remain on the plant surface for several weeks after application to be economical. Rainfall quickly washes Slam Insecticide from the plant. As a result of our work, Slam Insecticide was reformulated to include gluten to resist wash-off by rainfall, thus extending the residual activity of the application. Following this work, cooperation with an industry partner led to the development and commercialization of a spray adjuvant to improve control of corn rootworm (CRW) with one-tenth the normal amount of insecticide. This adjuvant, Cidetrak CRW, is currently available to growers. Another technology developed through this project, was the Fruitsphere for control of fruit flies in orchards. In a cooperative project with university scientists, fruit mimicking spheres composed of corn flour, corn syrup, and sugar with a coating in insecticide continues to progress toward commercialization.
Current efforts are directed at using biodegradable starch-based plastics to mold the fruit mimic. Recent formulation and stabilization research has led to the development of a spray-dried formulation process for imparting solar stability to spores of the bioinsecticidal fungus B. bassiana for improved persistence under field conditions. 6. What do you expect to accomplish, year by year, over the next 3 years? During FY 2004, studies will continue on optimizing production and stabilization methods for the bioherbicide Mycoleptodiscus terrestris (MT) . Bioassays and field trials with formulated MT preparations produced during these optimization studies will be evaluated for efficacy in controlling the invasive, aquatic weed, hydrilla. A new thrust will involve development of formulations for aquatic pest control. Production and formulation methodology for the bioherbicide M. terrestris will be standardized and registration of this agent will be pursued in collaboration with an
industrial partner. Work with the bioinsecticidal fungus P. fumosoroseus will focus on the development of formulates of this fungus useful in controlling the Formosan subterranean termite. Production of dried spore preparations for field-testing in New Orleans, LA, will continue. Related to microbial pesticides, we will work to develop techniques to produce novel formulations for fungal-based bioinsecticides. We will continue research on viral pesticides for control of mosquitoes in cooperation with ARS scientists and for control of velvet bean caterpillar with Brazilian scientists. In addition, research will be directed toward formulations of insect-active volatile compounds such as pheromones and chemicals from host plants. These new products will provide sustained release of attractants for use in various pest control strategies. During FY 2005, research will be directed toward formulations of insect active volatile compounds such as pheromones and chemicals from host plants.
These new products will provide sustained release of attractants for use in various pest control strategies. Work will continue on developing formulations that enhance the solar stability of insect pathogenic fungi for use as biopesticides and promising formulations will be tested under field conditions. For FY 2006, current work is expected to result in new formulations capable of protecting insect pathogenic fungal conidia. We will continue research on solar stability of insect pathogenic fungi and organize field-testing of these formulations. Also, efforts will be directed to identifying commercial partners to license these formulations. In FY-2004, a new Project Plan will be developed and reviewed under National Program 304. 7. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and
durability of the technology products? During 2003, an industrial partner has continued pursuit of an exclusive license on the ARS process for producing and stabilizing blastospores of the bioinsecticidal fungus Paecilomyces fumosoroseus (U.S. Patent #08,968, 808, Methods and Compositions for Producing Desiccation Tolerant Paecilomyces fumosoroseus Spores) and on the use of P. fumosoroseus for control of the Formosan subterranean termite (U.S. Patent Application S/N 09/989/287, Use of Paecilomyces spp. as Pathogenic Agents Against Subterranean Termites). It is envisioned that a P. fumosoroseus-based biopesticide may be available as commercial products for use in agricultural and urban markets within 3-5 years. Using our media optimization strategies for developing liquid production media for biopesticides, we continue collaborating with industrial and academic scientists in the United States and Mexico on developing production media for various biopesticides. This work has led to a
Specific Cooperative Agreement with academic scientists in Mexico to develop low-cost commercial methods for producing and formulating fungal biopesticides. We have also initiated research collaboration with Corps of Engineer scientists to develop production and stabilization methods for biopesticides used in controlling aquatic weeds. In addition, we have entered into a Cooperative Research and Development Agreement (CRADA) with a leading aquatic weed control company to develop methods for producing and formulating the biopesticide Mycoleptodiscus terrestris for control of hydrilla. This collaboration has resulted in a patent (Mycoherbicidal Compositions and Methods of Preparing and Using the Same, U.S. Patent #6,569,807) on the use of microsclerotia of MT for hydrilla control, jointly developed by ARS and the U.S. Army Corps of Engineers. Licensing of this patent is being pursued by our CRADA partner.
Impacts
(N/A)
Publications
- WRIGHT, M.S., JACKSON, M.A., CONNICK, W. J. POTENTIAL USE OF PAECILOMYCES FUMOSOROSEUS FOR CONTROL OF THE FORMOSAN SUBTERRANEAN TERMITE COPTOTERMES FORMOSANUS SHIRAKI. SOCIETY FOR INVERTEBRATE PATHOLOGY. 2003. v. 1: ABSTRACT p. 104.
- JACKSON, M.A. CULTURE MEDIA DESIGN FOR FERMENTATION PROCESSES. BIOTECHNOLOGY PROCESSES WORKSHOP, UNIVERSIDAD AUTONOMAS DE NUEVO LEON, MONTERREY, MEXICO. 2002. v. 1: ABSTRACT p. 34.
- KARGALIOGLU, Y., JACKSON, M.A. LIQUID CULTURE FERMENTATION PRODUCTION OF MICROSCLEROTIA OF MYCOLEPTODISCUS TERRESTRIS AS A BIOHERBICIDE FOR CONTROL OF HYDRILLA VERTICILLATA. AMERICAN SOCIETY FOR MICROBIOLOGY. 2003. v. 1: ABSTRACT p. 470.
- BEHLE, R. W. FORMULATIONS OF BIOPESTICIDES. BIOTECHNOLOGY PROCESSES WORKSHOP, UNIVERSIDAD AUTONOMAS DE NUEVO LEON, MONTERREY, MEXICO. 2002. v. 1: ABSTRACT p. 40.
- BEHLE, R.W. FORMULATIONS TO IMPROVE BACULOVIRUS INSECTICIDES. PROCEEDINGS OF SICONBIOL. 2003. v. 1: ABSTRACT p. 43.
- LELAND, J.E., BEHLE, R.W. DEVELOPMENT OF MYCOINSECTICIDES FOR CONTROLLING TARNISHED PLANT BUGS ON WILD HOST PLANTS. NATIONAL COTTON COUNCIL BELTWIDE COTTON CONFERENCE. 2003. v. 1: PROCEEDINGS P. 954-965.
- JACKSON, M.A., CLIQUET, SL., ITEN, L.B. MEDIA AND FERMENTATION PROCESSES FOR THE RAPID PRODUCTION OF HIGH CONCENTRATIONS OF STABLE BLASTOSPORES OF THE BIOINSECTICIDAL FUNGUS PAECILOMYCES FUMOSOROSEUS. BIOLOGICAL SCIENCE AND TECHNOLOGY. 2003. v. 13, p. 23-33.
- SHEARER, J.F., JACKSON, M.A. MYCOHERBICIDAL COMPOSITIONS AND METHODS OF PREPARING AND USING THE SAME. 2003. U.S. PATENT 6,569,807.
- SHAPIRO, D.I., BEHLE, R.W., MCGUIRE, M.R., LEWIS, E.E. FORMULATED ARTHROPOD CADAVERS FOR PEST SUPPRESSION. 2003. U.S. PATENT 6,524,601.
- VEGA, F.E., JACKSON, M.A., MERCADIER, G., POPRAWSKI, T.J. THE IMPACT OF NUTRITION ON SPORE YIELDS FOR VARIOUS ENTOMOPATHOGENS IN LIQUID CULTURE. WORLD JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY. 2003. v. 19. p. 363-368.
- WRIGHT, M.S., JACKSON, M.A., CONNICK, W.M. USE OF PAECILOMYCES SPP. AS PATHOGENIC AGENTS FOR CONTROL OF TERMITES. AMERICAN SOCIETY FOR MICROBIOLOGY. 2003. v. 1: ABSTRACT p. 439.
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Progress 10/01/01 to 09/30/02
Outputs
1. What major problem or issue is being resolved and how are you resolving it? Commercial interest in using living microbial agents for controlling agricultural pests such as weeds, insects, and plant pathogens or as probiotic feed additives for livestock is driven by an increasing public demand for environmentally-friendly, sustainable agricultural practices. The widespread use of chemical pesticides and antibiotic feed additives are often viewed as non-sustainable agricultural practices. An integrated pest management (IPM) approach using biological control agents (biopesticides) in conjunction with chemical pesticides is recognized as a sustainable pest management system which reduces chemical pesticide usage and deters the development of pest resistence to the chemical control. Likewise, the use of living microorganisms as feed additives (probiotics) has been shown to improve animal health, weight gain, and feed conversion efficiency. Effective probiotic feed
additives can reduce or eliminate the use of antibiotics in feeds. A significant constraint to the commercial use of living microbial agents in agriculture is the lack of low-cost methods for producing and formulating stable, effective microbial products. While living microbial agents can be produced using solid-substrate or liquid culture production methods, liquid culture fermentation processes are generally the most cost- effective. Optimization of deep-tank, liquid fermentation processes for the production of living microbial agents must take into account not only product yield but also microbial stability and product efficacy. Likewise, formulations for these microbial products must result in a stabilized, effective biopesticides in a form that is conducive to application technology commonly used in agriculture. Our four major research goals will be to (1) develop low-cost, deep-tank fermentation methods for the production of stable, effective living microbial propagules for
use as biocontrol agents or probiotic feed additives, (2) develop methods for drying and formulating fungal and bacterial microbial biopesticides that enhance the viability and efficacy of these agents during storage and application, respectively, (3) identify and develop potential bacterial or fungal biocontrol agents for the control of Fusarium graminearum, the causative agent of head scab of wheat and (4) develop bait and kill formulations for use with existing chemical and biological pesticides to decrease the amount of toxicant required for effective control. 2. How serious is the problem? Why does it matter? Public concerns over the widespread use of chemical pesticides and over increases in pest resistence to safe chemical controls has heightened interest in developing environmentally friendly pest control measures. The use of microbial pathogens as biopesticides for controlling weeds, insects and plant diseases is an acceptable alternative control measure. Needs for microbial
products that control plant pests remain high. For instance, viable control measures are currently unavailable for Fusarium head blight of wheat and barley which is estimated to cause more than 3 billion dollars in the 1990's. Likewise, while plant and insect pathologists have identified hundreds of microbial agents over the past 20 years that show good potential for use as biopesticides, less than 15 living microbial biopesticides have been commercially developed. In large part, the limited success in commercializing these microbial agents is due to a lack of economical production and formulation processes that deliver microbial products with a reasonable shelf-life and that provide consistent pest control under field conditions. These technical constraints to the use of biopesticides require research designed to understand how the nutritional and environmental conditions present during the production and formulation processes affect biopesticide yield, stability, and biocontrol
efficacy. Unless these problems are addressed, the widespread use of biopesticides will not become a reality. Likewise, the use of antibiotic feed additives has been implicated in hastening the development of antibiotic-resistant bacteria thus mitigating the usefulness of various classes of antibiotics for human and animal health uses. Many countries in Europe have already banned the prophylactic use of antibiotics in animal feeds. In many cases, living microbial feed additives have been shown to provide health and weight gain benefits comparable to those seen with antibiotic supplementation. Since the consistent efficacy of these products is dependant on the delivery of a living microbial feed additive, many of the technical constraints applicable to biopesticides are also affecting the commercial availability of probiotics. The widespread use of probiotic feed additives will require the development of low-cost production methods which yield stable, effective microbial feed
additives. 3. How does it relate to the national Program(s) and National Program Component(s) to which it has been assigned? This research focuses on the development of cost effective fermentation and formulation processes for mass producing microbial biopesticides that contributes to National Program 304 "Crop Protection and Quarantine" (70%) . The expected outcome from this research is the commercial use of microbial biopesticides and probiotic feed additives which will lead to additional markets for the agricultural commodities used as nutrients in the fermentation and formulation processes thus contributing to National Program 306 "Quality and Utilization of Agricultural Products" (30%). In addition, the use of biopesticides will reduce pest damage to plant products thereby improving product quality and marketability. The research directed at discovering and developing microorganisms for biologically controlling head scab of wheat demonstrates a strong association with the
Biological Control component of National Program 303 "Plant Diseases." 4. What was your most significant accomplishment this past year? A. Single Most Significant Accomplishment During FY 2002: Very few effective chemical or biological pesticides are available for controlling the Formosan subterranean termite, an invasive insect species that is causing devastating problems in the Southern United States. In collaboration with ARS scientists Maureen Wright and William Connick, Jr., at the Southern Regional Research Center in New Orleans, LA, dried preparations of the bioinsecticidal fungus Paecilomyces fumosoroseus (Pfr) , manufactured using ARS' patented production technology, were evaluated as a potential biological control agent against the Formosan subterranean termite. Formosan termites exposed to Pfr spore preparations were not repelled by the formulation, were infected and killed by the fungus and transmitted the Pfr infection to unexposed nestmates. The development of P.
fumosoroseus as an effective, environmentally friendly, nonchemical control measure for the Formosan termite will help reduce the estimated $1 billion annual damage caused by this termite. B. Other Significant Accomplishment(s), if any: A critical step in producing a commercially available biocontrol product is devising procedures for stabilizing biomass of the biological agent while maintaining product effectiveness, yet nothing is known regarding accomplishing this goal with biocontrol agents active against Fusarium head blight of wheat and barley. At the National Center for Agricultural Utilization Research laboratory in Peoria, IL, we discovered a new cryoprotectant compound, melezitose (a sugar composed of two glucose and one fructose molecules), that showed superiority over seven other previously described cryoprotectants in preserving the viability of freeze-dried cells of our superior Fusarium head blight antagonist, Cryptococcus nodaensis OH 182.9. This compound was
effective in maintaining the viability of yeast OH 182.9 for more than 6 weeks when added at the beginning or end of cell production in liquid culture shake flask studies and was also effective when added to the yeast cells after producing them in 100-L liquid culture fermentors. This discovery is a significant step towards producing an effective, commercially available biological control product from our patent pending antagonist of Fusarium head blight of wheat. Solar irradiation is a major environmental constraint to the persistence of fungal and bacterial biological control agents after application. Formulation ingredients and processing conditions for spray drying were evaluated for encapsulating spores of the bioinsecticidal fungus Beauveria bassiana for protection from solar degradation and subsequent loss of activity. Key formulation ingredients and physical conditions for spray-drying encapsulated formulations of fungal spores were discovered that yielded viable fungal
spores with improved solar stability. The development of this spray drying processes for formulating fungal spores with improved solar stability will likely improve the biocontrol efficacy of these pest control agents under field conditions thereby enhancing their potential for commercial use in agriculture. C. Significant Accomplishments/Activities that Support Special Target Populations: none. D. Progress Report: Over the past year, research was conducted to develop formulations for the insect biocontrol fungus, Beauveria bassiana, and a mosquito-active virus for control of the mosquito vector of West Niles Fever. Work continues on evaluating microbial antagonists for controlling Fusarium head blight (scab) of wheat for amenability to commercial production and stabilization processes and biocontrol efficacy under field conditions. Nutritional and environmental conditions were optimized for the production and stabilization of the fungal bioherbicide Mycoleptodiscus terrestris, a
fungal pathogen and potential biocontrol agent for the aquatic weed hydrilla. Results from these studies yielded a method for producing high concentrations of a stable, infective form of the fungus. In collaboration with an industrial partner, this fungal bioherbicide is being field tested in Texas and Florida for hydrilla control. 5. Describe your major accomplishments over the life of the project, including their predicted or actual impact? While this project is in its third year, it builds on our previous project 3620-41000-057-00D, entitled "Production and stabilization of microbial bioherbicides." This previous research led to the development of deep-tank fermentation methods for the production of three promising microbial biopesticides; Colletotrichum truncatum, a fungal biopesticide for control of the weed hemp sesbania, Xanthomonas campestris, a bacterial biopesticide used for controlling the weedy grass, Poa annua (annual bluegrass), and Paecilomyces fumosoroseus, a fungal
pathogen of numerous insect pests including the silverleaf whitefly. In these studies, the nutritional composition of liquid culture production media were optimized to maximize yields, propagule stability, and biocontrol efficacy. This novel research showed that the diets fed microbes during the production process could have a profound influence on the quantity and quality of the biopesticidal product. A major discovery derived from project 3620-41000-057 was the development and patenting of a method for producing infective, stable spores of the bioinsecticide Paecilomyces fumosoroseus (U.S. Patent #5,968,808, October 19, 1999). Work with the production and stabilization of P. fumosoroseus spores continues to be a major focus of the present project. It is expected that our P. fumosoroseus spore production process will become a commercial method for producing this promising bioinsecticide. A novel approach to producing biopesticidal spores of P. fumosoroseus is on-site production
in portable fermentation equipment. In order to use portable fermentation equipment for the on-site production and delivery of spores of P. fumosoroseus, process conditions were developed which favor the rapid production of P. fumosoroseus spores while inhibiting the growth of undesirable microbes. High concentrations of P. fumosoroseus spores were rapidly produced in a portable fermentor with little or no bacterial contamination when the pH of the production medium was lowered and appropriate nutrient concentrations were provided in the production medium. These studies have demonstrated that the on-site production of fungal biocontrol agents is feasible and, specifically, that high concentrations of P. fumosoroseus can be rapidly produced using an aseptic, portable fermentation system. Other insect targets for the bioinsecticide P. fumosoroseus have been identified including the Formosan subterranean termite and the Mexican bean beetle. In laboratory and field studies conducted at
the NCAUR in Peoria, IL, and in field studies conducted at Wooster, OH, biocontrol microbes were isolated and tested which greatly reduced the severity of scab of wheat, scab disease incidence, and mycotoxin content of wheat. The most effective microbes reduced disease severity in field trials by as much as 75% percent with disease reduction being demonstrated on 3 different types of wheat. Significantly, the microorganisms we discovered reduced scab of wheat disease symptoms when the microbes were grown in a liquid culture medium that would be affordable for use on a commercial scale. Patent protection has been obtained for 7 microbial isolates which have shown excellent biocontrol efficacy in controlling scab of wheat. The eventual development of these agents as biopesticides for scab of wheat will provide farmers with a critically needed tool for reducing the devastating impact of this disease. Project 3620-41000-070-00D entitled "Formulation and adjuvants for biological
controls and insect attractants" was adsorbed into 3620-22410- 004-00D in 2000. Over the life of this incorporated project, several formulations and adjuvants have been developed and tested for use with microbial pesticides and for use in reducing the amounts of chemical pesticides needed for control. Corn flour, gluten, and lignin were all tested as adjuvants for reducing wash-off by rain and for preventing UV light from inactivating microbial pesticides. Three patents have been issued concerning these formulations. Prior to 2000, adjuvants were tested with a product called Slam Insecticide, which is being used to control adult corn rootworms with a very low amount of insecticide. Slam must remain on the plant surface for several weeks after application to be economical. Rainfall quickly washes Slam from the plant. As a result of our work, Slam was reformulated to include gluten to resist wash-off by rainfall, thus extending the residual activity of the application. Following
this work, cooperation with an industry partner lead to the development and commercialization of a spray adjuvant to improve control of corn rootworms with one tenth the normal amount of insecticide. This adjuvant, Cidetrak CRW, is currently available to growers. Another technology developed through this project, was the Fruitsphere for control of fruit flies in orchards. In a cooperative project with university scientists, a fruit mimicking sphere composed of corn flour, corn syrup, and sugar with a coating in insecticide continues to progress toward commercialization. This device provides control of an important group of insect pests with a 1,000-fold reduction in the amount of insecticide by taking advantage of the natural behavior of the pest to be attracted to a fruit shaped device. Recent formulation and stabilization research has led to the development of a spray-dried, formulation process for imparting solar stability to spores of the bioinsecticidal fungus B. bassiana
for improved persistence under field conditions. 6. What do you expect to accomplish, year by year, over the next 3 years? During FY 2003, in association with fermentation studies, molecular biology approaches will be utilized to determine the mechanism(s) utilized by successful microbes in reducing the severity of scab disease. Studies will continue on optimizing production and stabilization methods for the bioherbicide Mycoleptodiscus terrestris (MT). Bioassays and field trials with formulated MT preparations produced during these optimization studies will be evaluated for efficacy in controlling the invasive, aquatic weed, hydrilla. A new thrust will involve development of formulations for aquatic pest control. Production and formulation methodology for the bioherbicide M. terrestris will be standardized and registration of this agent will be pursued in collaboration with an industrial partner. Related to microbial pesticides, we will work to develop techniques to produce novel
formulations for fungal-based bioinsecticides. We will continue research on viral pesticides for control of mosquitoes in cooperation with ARS scientists and with Brazilian scientists. In addition, research will be directed toward formulations of insect active volatile compounds such as pheromones and chemicals from host plants. These new products will provide sustained release of attractants for use in various pest control strategies. During FY 2004, biocontrol products produced using optimized drying and formulation protocols will be field tested. Chemostat studies in liquid culture with two or more agents with biocontrol potential will be conducted in laboratory studies to determine if stable biocontrol agent complexes of more than 1 strain can be discovered that have improved efficacy over that exhibited by single strains. A new thrust will involve development of formulations of both weed and insect pests in aquatic environments. Also, research will be directed toward
formulations of insect active volatile compounds such as pheromones and chemicals from host plants. These new products will provide sustained release of attractants for use in various pest control strategies. Work will continue on developing formulations that enhance the solar stability of insect pathogenic fungi for use as biopesticides and promising formulations will be tested under field conditions. For FY 2005, biocontrol products produced using optimized drying and formulation protocols will be field tested for control of Fusarium head blight of wheat. Chemostat studies in liquid culture with two or more agents with biocontrol potential will be conducted in laboratory studies to determine if stable biocontrol agent complexes of more than one strain can be discovered that have improved efficacy over that exhibited by single strains. Current work is expected to result in new formulations capable of protecting insect pathogenic fungal conidia. We will continue research on solar
stability of insect pathogenic fungi and organize field- testing of these formulations. Also, efforts will be directed to identifying commercial partners to license these formulations. 7. What technologies have been transferred and to whom? When is the technology likely to become available to the end user (industry, farmer other scientist)? What are the constraints, if known, to the adoption durability of the technology? During 2002, an industrial partner has applied for and is pursuing exclusive licenses on ARS's process for producing and stabilizing blastospores of the bioinsecticidal fungus Paecilomyces fumosoroseus (U.S. Patent #08,968,808 "Methods and Compositions for Producing Desiccation Tolerant Paecilomyces fumosoroseus Spores") and on the use of P. fumosoroseus for control of the Formosan subterranean termite ("Use of Paecilomyces spp. as pathogenic agents against subterranean termites" U.S. Patent Application S/N 09/989,287). It is envisioned that a P.
fumosoroseus-based biopesticides may be available as commercial products for use in agricultural and urban markets within 3-5 years. Using our media optimization strategies for developing liquid production media for biopesticides, we continue collaborating with industrial and academic scientists in the United States, France, Spain, Japan, and Mexico on developing production media for various biopesticides. This work has led to a Specific Cooperative Agreement with academic scientists in Mexico to develop low-cost commercial methods for producing and formulating fungal biopesticides. We have also initiated research collaboration with Army Corps of Engineer scientists to develop production and stabilization methods for biopesticides used in controlling aquatic weeds. In addition, we have entered into a Cooperative Research and Development Agreement (CRADA) with a leading aquatic weed control company to develop methods for producing and formulating the biopesticide Mycoleptodiscus
terrestris for control of hydrilla. Patents are pending on 7 microbial strains that are superior in reducing the severity of wheat head scab. This greatly increases the likelihood of commercial interest in developing a biological control product from one or more of these strains. Results of our successful discovery and field testing of biocontrol microbes effective against head scab of wheat have been presented at forums and scientific meetings attended by wheat society members, farmers, agricultural companies, and scientists. The application of this technology toward controlling scab on durum wheats has been successfully tested as part of a CRADA and the results have been conveyed to growers cooperative for Durum wheats. As part of research supported by the U.S. Wheat and Barley Scab Initiative, one of our superior yeast biocontrol strains was tested, with great success to date, at state agricultural experiment stations across the U.S. and wheat plots that were treated with this
strain shown to visiting agricultural workers at field days at many of these stations. Two option licenses were granted in 2000, a corn rootworm feeding stimulant and apple decoy technology for the directed delivery of pesticides. The licensee of the corn rootworm feeding stimulant is commercially producing this adjuvant, and the apple decoy technology licensed in 2000 is awaiting EPA registration.
Impacts
(N/A)
Publications
- Behle, R.W. Consumption of residue containing cucurbitacin feeding stimulant and reduced rates of carbaryl insecticide by western corn rootworm (Coleoptera: Chrysomelidae). Journal of Economic Entomology. 2001. v. 94. p. 1428-1433.
- Jackson, M.A., Schisler, D.A. Selecting fungal biocontrol agents amenable to production by liquid culture fermentation. Proceedings Seventh Biocontrol Working Group Meeting "Influence of A-Biotic and Biotic Factors on Biocontrol Agents." IOBC wprs Bulletin. 2002. p. 387-391.
- Schisler, D.A., Khan, N.I., Iten, L.B., Boehm, M.J. USDA-ARS, Ohio State University cooperative research on biologically controlling Fusarium head blight pilot-plant-scale production and processing of biomass of yeast antagonists. Proceedings 2001 National Fusarium Head Blight Forum. 2001. p. 87-90.
- Behle, R.W., Dowd, P.F., Tamez-Guerra, P., Lagrimini, L.M. Effect of transgenic plants expressing high levels of a tobacco anionic peroxidase on the toxicity of Anagrapha falcifera nucleopolyhedrovirus to Helicoverpa zea (Lepidoptera: Noctuidae). Journal of Economic Entomology. 2002. v. 95. p. 81-88.
- Wright, M.S., Connick, W.J., Jr., Jackson, M.A. Use of Paecilomyces spp. as pathogenic agents against subterranean termites. 2001. U.S. Patent Application S/N 09/989,287.
- Schisler, D.A., Khan, N.I, Boehm, M.J. Biological control of Fusarium head blight of wheat and deoxynivalenol levels in grain via use of microbial antagonists. DeVries, J.W., Trucksess, M.W., Jackson, L.S., editors. Kluwer Academic/Plenum Publishers, New York. Mycotoxins and Food Safety. 2002. p. 53-69.
- Shapiro-Ilan, D., Behle, R.W., McGuire, M.R., Lewis, E.E. Formulated arthropod for pest suppression. 2001. U.S. Patent Application S/N 09/875, 972.
- Sandoval-Coronado, C.F., Luna-Olvera, H.A., Arevalo-Nino, K., Jackson, M.A. , Poprawski, T.J., Galan-Wong, L.J. Drying and formulation of blastospores of Paecilomyces fumosoroseus (Hyphomycetes) produced in two different liquid media. World Journal of Microbiology and Biotechnology. 2001. v. 17. p. 423-428.
- Wraight, S.P., Jackson, M.A. Recent advances in production and formulation of entomopathogenic fungi. Society for Invertebrate Pathology. 2001. Abstract p. 81.
- Garcia-Guitierrez, C., Tamez-Guerra, P., McGuire, M.R., Jackson, M., Behle, R.W. Toxicity of Paecilomyces fumosoroseus produced in liquid and solid medium using a dipped leaf and topical bioassay on larvae of Mexican bean beetle (Coleoptera: Coccinellidae). Society for Invertebrate Pathology. 2001. Abstract p. 25.
- Khan, N.I., Schisler, D.A., Boehm, M.J., Slininger, P.J., Bothast, R.J. Selection and evaluation of microorganisms for biocontrol of Fusarium head blight of wheat incited by Gibberella zeae. Plant Disease. 2001. v. 85. p. 1253-1258.
- Shapiro-Ilan, D.I., Lewis, E.E., Behle, R.W., McGuire, M.R. Formulation of entomopathogenic nematode-infected cadavers. Journal of Invertebrate Pathology. 2001. v. 78. p. 17-23.
- Jackson, M.A. Production of Paecilomyces fumosoroseus blastospores. Proceedings of XXXIII Mexican Congress of Microbiology. 2002. p. 162.
- Pingel, R.L., Behle, R.W., McGuire, M.R., Shasha, B.S. Improvement of the residual activity of a cucurbitacin-based adult corn rootworm insecticide. Journal of Entomology Science. 2001. v. 36. p. 416-425.
- McGuire, M.R., Tamez-Guerra, P., Behle, R.W., Streett, D.A. Comparative field stability of selected entomopathogenic virus formulations. Journal of Economic Entomology. 2001. v. 94. p. 1037-1044.
- Tamez-Guerra, P., McGuire, M.R., Behle, R.W., Shasha, B.S., Pingle, R.L. Storage stability of Anagrapha falcifera nucleopolyhedroviruses (AfMNPV) as spray-dried formulations. Journal of Invertebrate Pathology. 2002. v. 79. p. 7-16.
- Jackson, M.A., Connick, W.J, Erhan, S.M., Vega, F.E., Cliquet, S., Payne, A.R. Factors affecting the desiccation tolerance and storage stability of blastospores of Paecilomyces fumosoroseus. Society for Invertebrate Pathology. 2001. Abstract p. 38.
- Schisler, D.A., Jackson, M.A., Slininger, P.J. Discovering, developing and deploying biological control agents. American Chemical Society. 2002. Abstract p. IEC323.
- Shearer, J.F., Jackson, M.A. Partnering to develop an endemic fungal pathogen as a bioherbicide for management of Hydrilla verticillata. Aquatic Plant Management Society. 2001. v. 41. Abstract p. 29.
- Schisler, D.A., VanCauwenberge, J.E. Discovery and scale-up of freeze- drying protocols for biomass of Fusarium head blight antagonist Cryptococcus nodaensis OH 182.9 (NRRL Y-30216). Phytopathology. 2002. v. 92(6). Abstract p. S73.
- Stelinski, L.L., Liburd, O.E., Wright, S., Prokopy, R.J., Behle, R., McGuire, M.R. Comparison of neonicotinoid insecticides for use with biodegradable and wooden spheres for control of key Rhagoletis species (Diptera: Tephritidae). Journal of Economic Entomology. 2001. v. 94. p. 1142-1150.
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Progress 10/01/00 to 09/30/01
Outputs
1. What major problem or issue is being resolved and how are you resolving it?
Commercial interest in using living microbial agents for controlling agricultural pests such as weeds, insects and plant pathogens or as probiotic feed additives for livestock is driven by an increasing public demand for environmentally-friendly, sustainable agricultural practices. The widespread use of chemical pesticides and antibiotic feed additives are often viewed as non-sustainable agricultural practices. An integrated pest management (IPM) approach using biological control agents (biopesticides) in conjunction with chemical pesticides is recognized as a sustainable pest management system which reduces chemical pesticide usage and deters the development of pest resistence to the chemical control. Likewise, the use of living microorganisms as feed additives (probiotics) has been shown to improve animal health, weight gain, and feed conversion efficiency. Effective probiotic feed additives can reduce or eliminate the use of antibiotics in feeds. A significant constraint to
the commercial use of living microbial agents in agriculture is the lack of low-cost methods for producing and formulating stable, effective microbial products. While living microbial agents can be produced using solid-substrate or liquid culture production methods, liquid culture fermentation processes are generally the most cost-effective. Optimization of deep-tank, liquid fermentation processes for the production of living microbial agents must take into account not only product yield but also microbial stability and product efficacy. Likewise, formulations for these microbial products must result in a stabilized, effective biopesticides in a form that is conducive to application technology commonly used in agriculture. Our four major research goals will be to (1) develop low-cost, deep-tank fermentation methods for the production of stable, effective living microbial propagules for use as biocontrol agents or probiotic feed additives, (2) develop methods for drying and
formulating fungal and bacterial microbial biopesticides that enhance the viability and efficacy of these agents during storage and application, respectively, (3) identify and develop potential bacterial or fungal biocontrol agents for the control of Fusarium graminearum, the causative agent of head scab of wheat and (4) develop bait and kill formulations for use with existing chemical and biological pesticides to decrease the amount of toxicant required for effective control.
2. How serious is the problem? Why does it matter?
Public concerns over the widespread use of chemical pesticides and over increases in pest resistence to safe chemical controls has heightened interest in developing environmentally friendly pest control measures. The use of microbial pathogens as biopesticides for controlling weeds, insects and plant diseases is an acceptable alternative control measure. Needs for microbial products that control plant pests remain high. For instance, viable control measures are currently unavailable for Fusarium head blight of wheat and barley which is estimated to cause more than 3 billion dollars in the 1990's. Likewise, while plant and insect pathologists have identified hundreds of microbial agents over the past 20 years that show good potential for use as biopesticides, less than 15 living microbial biopesticides have been commercially developed. In large part, the limited success in commercializing these microbial agents is due to a lack of economical production and formulation processes
that deliver microbial products with a reasonable shelf-life and that provide consistent pest control under field conditions. These technical constraints to the use of biopesticides require research designed to understand how the nutritional and environmental conditions present during the production and formulation processes affect biopesticide yield, stability and biocontrol efficacy. Unless these problems are addressed, the widespread use of biopesticides will not become a reality. Likewise, the use of antibiotic feed additives has been implicated in hastening the development of antibiotic-resistant bacteria thus mitigating the usefulness of various classes of antibiotics for human and animal health uses. Many countries in Europe have already banned the prophylactic use of antibiotics in animal feeds. In many cases, living microbial feed additives have been shown to provide health and weight gain benefits comparable to those seen with antibiotic supplementation. Since the
consistent efficacy of these products is dependant on the delivery of a living microbial feed additive, many of the technical constraints applicable to biopesticides are also affecting the commercial availability of probiotics. The widespread use of probiotic feed additives will require the development of low-cost production methods which yield stable, effective microbial feed additives.
3. How does it relate to the National Program(s) and National Component(s)?
This research focuses on the development of cost effective fermentation and formulation processes for mass producing microbial biopesticides that contributes to National Program 304 "Crop and Commodity Pest Biology, Control and Quarantine" (70%). The expected outcome from this research is the commercial use of microbial biopesticides and probiotic feed additives which will lead to additional markets for the agricultural commodities used as nutrients in the fermentation process thus contributing to National Program 306 "New Uses, Quality and Marketability of Plant and Animal Products" (30%). In addition, the use of biopesticides will reduce pest damage to plant products thereby improving product quality and marketability. The research directed at discovering and developing microorganisms for biologically controlling head scab of wheat demonstrates a strong association with the Biological Control component of National Program 303 "Plant Disease".
4. What were the most significant accomplishments this past year?
A. Single Most Significant Accomplishment during FY 2001: Research was conducted to develop a commercial method for producing and stabilizing the fungal bioherbicide Mycoleptodiscus terrestris (MT) for control of hydrilla, a serious invasive, aquatic weed in the Southern United States. In collaboration with a U.S. Army Corps of Engineer scientist, a deep-tank fermentation process was developed at ARS's National Center for Agricultural Utilization Research for the production of MT. The MT product derived from this fermentation process survived drying and was highly infective when applied to hydrilla. These results have led to the development of a Cooperative Research and Development Agreement (CRADA) with SePRO Corporation, Carmel, IN, for the commercial development of MT as a bioherbicide for hydrilla control. B. Other Significant Accomplishment(s), if any: Farm losses due to scab of wheat have been conservatively estimated to exceed 3 billion dollars in the 1990's in the United
States alone with devastating losses occurring in the Upper Midwest states, Illinois, Indiana, and Ohio. Pilot-plant scale antagonist inoculum production (30 and 100 L fermentors) and processing protocols were developed at ARS's National Center for Agricultural Utilization Research in Peoria, IL, and the resultant antagonist inoculum evaluated for effectiveness against scab of wheat in a large number of field trials. Antagonist inoculum produced using these procedures effectively reduced scab of wheat both in greenhouse and in field trials reporting to date that were conducted at 16 sites across the United States as part of the U.S. Wheat and Barley Scab Initiative's uniform trial. The discovery of pilot-scale antagonist production and processing methodologies adds further evidence to the feasibility of developing a biological control product from one or more of our patent-pending microbial strains for use against scab of wheat. C. Significant Accomplishments/Activities that
Support Special Target Populations: Nothing to report. D. Progress Report: Over the past year, research was conducted to develop formulations for the insect biocontrol fungus, Beaveria bassiana, and a mosquito-active virus for control of the mosquito vector of West Niles Fever. These studies have resulted in the development of basic formulations of both biocontrol agents and of methodology for testing formulation stability and efficacy. In addition, testing of spore preparations of the bioinsecticidal fungus, Paecilomyces fumosoroseus, produced using ARS' patented production method have led to the discovery of other important insect targets for this bioinsecticide.
5. Describe the major accomplishments over the life of the project including their predicted or actual impact.
While this research project is in its second year, it builds on our previous project #3620-41000-057. These previous studies led to the development of deep-tank fermentation methods for the production of three promising microbial biopesticides; Colletotrichum truncatum, a fungal biopesticide for control of the weed hemp sesbania, Xanthomonas campestris, a bacterial biopesticide used for controlling the weedy grass, Poa annua (annual bluegrass) and Paecilomyces fumosoroseus, a fungal pathogen of numerous insect pests including the silverleaf whitefly. In these studies, the nutritional composition of liquid culture production media were optimized to maximize yields, propagule stability and biocontrol efficacy. This novel research showed that the diets fed microbes during the production process could have a profound influence on the quantity and quality of the biopesticidal product. A major discovery derived from project #3620-41000-057 was the development and patenting of a method
for producing infective, stable spores of the bioinsecticide Paecilomyces fumosoroseus (US patent #5,968,808, October 19, 1999). Work with the production and stabilization of P. fumosoroseus spores continues to be a major focus of the present research project. It is expected that our P. fumosoroseus spore production process will become a commercial method for producing this promising bioinsecticide. A novel approach to producing biopesticidal spores of P. fumosoroseus is on-site production in portable fermentation equipment. In order to use portable fermentation equipment for the on-site production and delivery of spores of P. fumosoroseus, process conditions were developed which favor the rapid production of P. fumosoroseus spores while inhibiting the growth of undesirable microbes. High concentrations of P. fumosoroseus spores were rapidly produced in a portable fermentor with little or no bacterial contamination when the pH of the production medium was lowered and appropriate
nutrient concentrations were provided in the production medium. These studies have demonstrated that the on-site production of fungal biocontrol agents is feasible and, specifically, that high concentrations of P. fumosoroseus can be rapidly produced using an aseptic, portable fermentation system. In laboratory and field studies conducted at the NCAUR in Peoria, IL, and in field studies conducted at Wooster, OH, biocontrol microbes were isolated and tested which greatly reduced the severity of scab of wheat, scab disease incidence and mycotoxin content of wheat. The most effective microbes reduced disease severity in field trials by as much as 75% percent with disease reduction being demonstrated on 3 different types of wheat. Patent protection is being sought for 7 microbial isolates which have shown excellent biocontrol efficacy in controlling scab of wheat. The eventual development of these agents as biopesticides for scab of wheat will provide farmers with a needed tool for
controlling this devastating disease. Since the last progress report, #3620- 41000-070-00D entitled "Formulation and adjuvants for biological controls and insect attractants" was adsorbed into this research project. Over the life of this incorporated project, several formulations and adjuvants have been developed and tested for use with microbial pesticides and for use in reducing the amounts of chemical pesticides needed for control. Corn flour, gluten, and lignin were all tested as adjuvants for reducing wash-off by rain and for preventing UV light from inactivating microbial pesticides. Three patents have been issued concerning these formulations. Prior to 2000, adjuvants were tested with a product called Slam Insecticide, which is being used to control adult corn rootworms with a very low amount of insecticide. Slam must remain on the plant surface for several weeks after application to be economical. Rainfall quickly washes slam from the plant. As a result of our work,
slam was reformulated to include gluten to resist wash-off by rainfall, thus extending the residual activity of the application. Following this work, cooperation with an industry partner lead to the development and commercialization of a spray adjuvant to improve control of corn rootworms with one tenth the normal amount of insecticide. This adjuvant, Cidetrak CRW, is currently available to growers. Another technology developed through this project, was the Fruitsphere for control of fruit flies in orchards. In a cooperative project with university scientists, a fruit mimicking spheres composed of corn flour, corn syrup, and sugar with a coating in insecticide continues to progress toward commercialization. This device provides control of an important group of insect pests with a 1000 fold reduction in the amount of insecticide by taking advantage of the natural behavior of the pest to be attracted to a fruit shaped device.
6. What do you expect to accomplish, year by year, over the next 3 years?
During FY 2002, the Pfr spore production and stabilization technology will be transferred to an industrial partner for commercialization of Pfr as a bioinsecticide. In collaboration with an industrial partner, large-scale field and greenhouse studies using Pfr spores, produced under commercial conditions, for control of various insects will be conducted to satisfy EPA registration requirements. In cooperation with partners, field tests will be conducted to control head scab of wheat with cells of biocontrol agents that were produced using the optimal liquid culture conditions discovered during FY 2001 research. These optimally produced biocontrol agents will then be field tested in combination with the best resistant varieties and small quantities of the best available fungicides to determine if these integrated disease management tools can be used to provide more effective control of scab disease. Studies designed to optimize production and stabilization methods for the
bioherbicide Mycoleptodiscus terrestris (MT) will be conducted. Bioassays and field trials with formulated MT preparations produced during these optimization studies will be evaluated for efficacy in controlling the invasive, aquatic weed, hydrilla. For the apple decoy project, commercial production and coating technology will be developed. Novel formulations for fungal- and viral-based bioinsecticides will be developed for B. bassiana for whitefly control and a baculovirus for mosquito control. During FY 2003, in association with fermentation studies, molecular biology approaches will be utilized to determine the mechanism(s) utilized by successful microbes in reducing the severity of scab disease. A new thrust will involve development of formulations for aquatic pest control. Production and formulation methodology for the bioherbicide MT will be standardized and registration of this agent will be sought in collaboration with an industrial partner. In addition, research will
be directed toward formulations of insect active volatile compounds such as pheromones and chemicals from host plants. These new products will provide sustained release of attractants for use in various pest control strategies. During FY 2004, biocontrol product produced using optimized drying and formulation protocols will be field tested. Chemostat studies in liquid culture with two or more agents with biocontrol potential will be conducted in laboratory studies to determine if stable biocontrol agent complexes of more than 1 strain can be discovered that have improved efficacy over that exhibited by single strains. Current work is expected to result in new formulations capable of protecting insect pathogenic fungal conidia. Work will continue on developing formulations that enhance the solar stability of insect pathogenic fungi for use as biopesticides and promising formulations will be tested under field conditions.
7. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end user (industry, farmer, other scientists)? What are the constraints if known, to the adoption & durability of the technology product?
Using our media optimization strategies for developing liquid production media for biopesticides, we are collaborating with industrial and academic scientists in the United States, France, Spain, Japan and Mexico on developing production media for various biopesticides. This work has led to a Specific Cooperative Agreements between ARS and academic scientists in Mexico to develop low-cost commercial methods for producing and formulating fungal biopesticides and between ARS and Corps of Engineer scientists to develop production and stabilization methods for biopesticides used in controlling aquatic weeds. In addition, ARS and SEPRO Corporation have entered into a Cooperative Research and Development Agreement (CRADA) to develop methods for producing and formulating the biopesticide Mycoleptodiscus terrestris for control of hydrilla. A U.S. patent was issued on ARS's process for producing and stabilizing blastospores of the bioinsecticidal fungus Paecilomyces fumosoroseus (U.S.
Patent #08,968,808 "Methods and Compositions for Producing Desiccation Tolerant Paecilomyces fumosoroseus Spores"). A CRADA partner is pursuing commercial development of this technology and has received an exclusive license on the patent. Foreign patent rights for this process are under prosecution in Europe, Canada and Mexico. It is envisioned that a P. fumosoroseus-based biopesticides may be available as commercial products within 3-5 years. Results of our successful discovery and field testing of biocontrol microbes effective against head scab of wheat have been presented at forums and scientific meetings attended by wheat society members, farmers, agricultural companies, and scientists. These include: The National Fusarium Head Blight Forum, National Association of Wheat Growers and the Annual Meeting of the American Phytopathological Society.
8. List your most important publications in the popular press (no abstracts) and presentations to non-scientific organizations and articles written about your work (NOTE: this does not replace your peer-reviewed publications which are listed below)
Yeast debuts in test on controlling wheat scab, Agricultural Research, USDA-ARS, June 2001. A Healthy History: National Center for Agricultural Utilization Research Marks 60 Years, Agricultural Research, USDA-ARS, July, 2001. Aarfusarium ligt wereldwijd onder vuur (Fusarium headblight currently under fire in the world). Boerderij/Akkerbouw (Farm and Crops), June 12, 2001. Government, Industry Team up Against Exotic Water Weed, ARS News and Information Services, USDA-ARS, April 26, 2001. (Electronic publication) Radio interview on biologically controlling wheat scab, radio station WMBD with Agribusiness radio personality Colleen Callahan, March 1, 2001.
Impacts
(N/A)
Publications
- Dowd, P.F., Pingle, R.L., Ruhl, D., Shasha, B.S., Behle, R.W., Penland, D.R., McGuire, M.R., Faron II, E.J. Multiacreage evaluation of aerially applied adherent malathion granules for selective insect control and indirect reduction of mycotoxigenic fungi in specialty corn. Journal of Economic Entomology. 2000. v. 93(5). p. 1424-1428.
- Pereyra-Alferez, B., Salazar-Macias, A.I., Damas-Buenrostro, L.C., Vazquez-Juarez, C., Jackson, M.A., Galan-Wong, L.J. Los Micoinsecticidas: Avances y alternativas. Badii, M.H., Flores, A.E., Galan-Wong, L.J., editors. Monterrey, Nuevo Leon, Mexico: Universidad Autonoma de Nuevo Leon. Fundamentos y Perspectivas de Control Biologico. 2000. p. 325-338.
- Ierulli, J.M., Schisler, D.A., Gessner, R.V. Potential of uridine augmentation to enhance wheat head colonization and efficacy of Fusarium Head Blight antagonist Bacillus subtilis AS 43.3. Phytopathology. 2001. v. 91:6 (suppl.): Abstract p. S42.
- Khan, N.I., Schisler, D.A., Boehm, M.J. Biocontrol of Fusarium head blight of wheat: Effect of Tween-80, culture age of antagonist and combining antagonists on disease suppression. Phytopathology. 2001. v. 91:6 (suppl.): Abstract p. S48.
- Schisler, D.A., Khan, N.I., Boehm, M.J. Reduction of Fusarium head blight disease severity and deoxynivalenol levels in grain via application of choline metabolizing antagonists at wheat flowering. Proceedings of the 220th ACS National Meeting, Division of Agricultural & Food Chemistry. 2000. Abstract No. 46.
- Wraight, S.P., Jackson, M.A., DeKock, S.L. Production, stabilization and formulation of fungal biocontrol agents. Butt, T., Jackson, C., Magan, N., editors. Wallingford, United Kingdom: CAB International; Fungal Biocontrol Agents - Progress, Problems and Potential. Chapter 10. 2001. p. 253-287.
- Vega, F.E., Mercadier, G., Jackson, M.A. Fungi associated with the coffee berry borer, Hypothenemus hampei. Smits, P.H., editor. Insect Pathogens and Insect Parasitic Nematodes. 2000. IOBC wprs Bulletin v. 23(2). p. 159.
- Schisler, D.A., Khan, N.I., Iten, L.B., Boehm, M.J. Scale-up of biomass production, processing and storage for two yeast antagonists of Gibberella zeae. Phytopathology. 2001. v 91:6 (suppl.): Abstract p. S80.
- Lee, F.N., Jackson, M.A. Walker, N.R. Characteristics of Pyricularia grisea "microsclerotia" produced in shake culture. Proceedings of the 28th Mtg. Rice Technical Working Group. 2000. p. 82.
- Schisler, D.A., Khan, N.I., Boehm, M.J., Lipps, P.E. USDA-ARS, Ohio State University cooperative research on biologically controlling Fusarium head blight. Field tests of antagonists in 2000. Proceedings of the National Fusarium Head Blight Forum. 2000. p. 105-108.
- Vega, F.E., Mercadier, G., Jackson, M.A. Screening fungal pathogens of insects for amenability to liquid culture spore production. XXI International Congress of Entomology. 2000. p. 545.
- Jackson, M.A. Selecting and producing biopesticides using liquid culture fermentation. Society for Industrial Microbiology. 2001. Abstract p. 64.
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Progress 10/01/99 to 09/30/00
Outputs
1. What major problem or issue is being resolved and how are you resolving it?
Commercial interest in using living microbial agents for controlling agricultural pests such as weeds, insects, and plant pathogens or as probiotic feed additives for livestock is driven by an increasing public demand for environmentally-friendly, sustainable agricultural practices. The widespread use of chemical pesticides and antibiotic feed additives are often viewed as non-sustainable agricultural practices. An integrated pest management (IPM) approach using biological control agents (biopesticides) in conjunction with chemical pesticides is recognized as a sustainable pest management system which reduces chemical pesticide usage and deters the development of pest resistence to the chemical control. Likewise, the use of living microorganisms as feed additives (probiotics) has been shown to improve animal health, weight gain, and feed conversion efficiency. Effective probiotic feed additives can reduce or eliminate the use of antibiotics in feeds. A significant constraint to
the commercial use of living microbial agents in agriculture is the lack of low-cost methods for producing stable, effective microbial products. While living microbial agents can be produced using solid-substrate or liquid culture production methods, liquid culture fermentation processes are generally the most cost-effective. Optimization of deep-tank, liquid fermentation processes for the production of living microbial agents must take into account not only product yield but also microbial stability and product efficacy. Our three major research goals will be to 1) develop low-cost, deep-tank fermentation methods for the production of stable, effective living microbial propagules for use as biocontrol agents or probiotic feed additives, 2) develop methods for drying and formulating fungal and bacterial microbial biopesticides which enhance the viability and stability of these agents during storage, and 3) identify and develop potential bacterial or fungal biocontrol agents for the
control of Fusarium graminearum, the causative agent of head scab of wheat.
2. How serious is the problem? Why does it matter?
Public concerns over the widespread use of chemical pesticides and over increases in pest resistence to these chemical controls have heightened interest in developing environmentally friendly pest control measures. The use of microbial pathogens as biopesticides for controlling weeds, insects, and plant diseases is an acceptable alternative control measure. Needs for microbial products that control plant pests remain high. For instance, viable control measures are currently unavailable for Fusarium head blight of wheat and barley which is estimated to cause more than 3 billion dollars in the 1990's. Plant and insect pathologists have identified hundreds of microbial agents which show good potential for use as biopesticides, yet, over the past 20 years, less than 15 living microbial biopesticides have been commercially developed. In large part, the limited success in commercializing these microbial agents is due to a lack of economical production methods, poor product shelf-life,
and inconsistent pest control under field conditions. Unless these problems are addressed, the widespread use of biopesticides will not become a reality. Research designed to understand how the nutritional and environmental conditions present during the production process affect biopesticide yield, stability, and biocontrol efficacy is needed if cost-effective production methods are to be developed. Likewise, the use of antibiotic feed additives has been implicated in hastening the development of antibiotic-resistant bacteria thus mitigating the usefulness of various classes of antibiotics for human and animal health uses. Many countries in Europe have already banned the prophylactic use of antibiotics in animal feeds. In many cases, living microbial feed additives have been shown to provide health and weight gain benefits comparable to those seen with antibiotic supplementation. Since the consistent efficacy of these products is dependant on the delivery of a living microbial
feed additive, many of the technical constraints applicable to biopesticides are also affecting the commercial availability of probiotics. The widespread use of probiotic feed additives will require the development of low-cost production methods which yield stable, effective microbial feed additives.
3. How does it relate to the National Program(s) and National Component(s)?
This research focuses on the development of cost effective fermentation processes for mass producing microbial biopesticides which contributes to National Program 304 "Crop and Commodity Pest Biology, Control and Quarantine" (70%). The expected outcome from this research is the commercial use of microbial biopesticides and probiotic feed additives which will lead to additional markets for the agricultural commodities used as nutrients in the fermentation process thus contributing to National Program 306 "New Uses, Quality and Marketability of Plant and Animal Products" (30%). In addition, the use of biopesticides will reduce pest damage to plant products thereby improving product quality and marketability. The research directed at discovering and developing microorganisms for biologically controlling head scab of wheat demonstrates a strong association with the Biological Control component of National Program 303 "Plant Disease."
4. What were the most significant accomplishments this past year?
A. Single Most Significant Accomplishment During FY 2000 Year: Alternative control methods for Fusarium head blight (scab) of wheat are needed as evidence by the fact that U.S. farm losses in the 1990's due to scab are conservatively estimated to exceed 3 billion dollars. Laboratory and field studies were conducted by ARS and Ohio State University scientists at ARS's National Center for Agricultural Utilization Research (NCAUR) in Peoria, IL, and on field plots in Wooster, OH, and Langdon, ND. This work demonstrated that the effectiveness of recently discovered microbial antagonists could be increased by changing the nutritional characteristics of the liquid culture production medium and that these biocontrol agents reduced the severity of scab of wheat and scab disease incidence in field trials by greater than 50% with disease reduction being demonstrated on three different types of wheat. With this discovery, another crucial step towards the development of a commercial
biological control product for use against Fusarium head blight (scab) of wheat has been completed. B. Other Significant Accomplishment(s), if any: The use of portable fermentors for on-site production of microbial biocontrol agents will help reduce biopesticide costs associated with the drying, packaging, shipping, and storing of these living microbial agents. In collaboration with our Cooperative Research and Development Agreement (CRADA) partner, methods were developed here at ARS's National Center for Agricultural Utilization Research to produce bioinsecticidal spores of the fungus Paecilomyces fumosoroseus using portable fermentation equipment. Protocols were developed for concentrating the spore production medium and for rehydrating and stabilizing the P. fumosoroseus spore inoculum for use in the portable fermentation equipment. These accomplishments support the use of portable fermentation equipment in the production of the fungal bioinsecticide Paecilomyces fumosoroseus.
C. Significant Accomplishments/Activities that Support Special Target Populations: Nothing to report.
5. Describe the major accomplishments over the life of the project including their predicted or actual impact.
While this CRIS project is in its second year, it builds on our previous CRIS project #3620-41000-057. These previous studies led to the development of deep-tank fermentation methods for the production of three promising microbial biopesticides: Colletotrichum truncatum, a fungal biopesticide for control of the weed hemp sesbania; Xanthomonas campestris, a bacterial biopesticide used for controlling the weedy grass; Poa annua (annual bluegrass) and Paecilomyces fumosoroseus, a fungal pathogen of numerous insect pests including the silverleaf whitefly. In these studies, the nutritional composition of liquid culture production media were optimized to maximize yields, propagule stability, and biocontrol efficacy. This novel research showed that the diets fed microbes during the production process could have a profound influence on the quantity and quality of the biopesticidal product. A major discovery derived from CRIS project #3620-41000-057 was the development and patenting of a
method for producing infective, stable spores of the bioinsecticide Paecilomyces fumosoroseus (U.S. Patent #5,968,808, October 19, 1999). Work with the production and stabilization of P. fumosoroseus spores continues to be a major focus of the present CRIS project. It is expected that our P. fumosoroseus spore production process will become a commercial method for producing this promising bioinsecticide. A novel approach to producing biopesticidal spores of P. fumosoroseus is on-site production in portable fermentation equipment. In order to use portable fermentation equipment for the on-site production and delivery of spores of P. fumosoroseus, process conditions were developed which favor the rapid production of P. fumosoroseus spores while inhibiting the growth of undesirable microbes. High concentrations of P. fumosoroseus spores were rapidly produced in a portable fermentor with little or no bacterial contamination when the pH of the production medium was lowered and
appropriate nutrient concentrations were provided in the production medium. These studies have demonstrated that the on-site production of fungal biocontrol agents is feasible and, specifically, that high concentrations of P. fumosoroseus can be rapidly produced using an aseptic, portable fermentation system. In laboratory and field studies conducted at the NCAUR in Peoria, IL, and in field studies conducted at Wooster, OH, biocontrol microbes were isolated and tested which greatly reduced the severity of scab of wheat, scab disease incidence, and mycotoxin content of wheat. The most effective microbes reduced disease severity in field trials by as much as 75% percent with disease reduction being demonstrated on three different types of wheat.
6. What do you expect to accomplish, year by year, over the next 3 years?
During FY 2001, it is expected that work with the biopesticide P. fumosoroseus (Pfr) will lead to the development of an improved commercial spore production medium which will be tested in large scale fermentors in collaboration with an industrial partner. The Pfr spore formulations and drying techniques currently being evaluated will be narrowed to one or two candidates. These formulations will be tested for biopesticide stability and efficacy in greenhouse and field studies in collaboration with industrial partners and ARS scientists. A commercial medium and fermentation protocol for producing Pfr spores using on-site fermentors will be optimized in collaboration with an industrial partner. Work on controlling wheat scab with microbial antagonists will identify optimal nutritional and chemical environments for the liquid culture production of microbial antagonist. Field trials with our most promising microbial antagonist through participation in the Uniform fungicide trials of
the U.S. Wheat and Barley Scab Initiative will demonstrate the effectiveness of these biocontrol agents in as many as six wheat growing locations across the U.S. During FY 2002, the Pfr spore production and stabilization technology will be transferred to an industrial partner for commercialization of Pfr as a bioinsecticide. In collaboration with an industrial partner, large scale field and greenhouse studies using Pfr spores for control of various insects will be conducted to satisfy EPA registration requirements. Field trials will be conducted in collaboration with an industrial partner to optimize on-site Pfr spore production and application in greenhouses and vegetable crops. In cooperation with partners, field tests will be conducted to control head scab of wheat with cells of biocontrol agents that were produced using the optimal liquid culture conditions discovered during FY 2001 research. These optimally produced biocontrol agents will then be field tested in combination
with the best resistant varieties and small quantities of the best available fungicides to determine if these integrated disease management tools can be used to provide more effective control of scab disease. During FY 2003, in association with fermentation studies, molecular biology approaches will be utilized to determine the mechanism(s) utilized by successful microbes in reducing the severity of scab disease.
7. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end user (industry, farmer, other scientists)? What are the constraints if known, to the adoption & durability of the technology product?
Using our media optimization strategies for developing liquid production media for biopesticides, we are collaborating with industrial and academic scientists in the U.S., France, Spain, Japan, and Mexico on developing production media for various biopesticides. This work has led to Specific Cooperative Agreements between ARS and academic scientists in Mexico to develop low-cost commercial methods for producing and formulating fungal biopesticides and between ARS and Corps of Engineer scientists to develop production and stabilization methods for biopesticides used in controlling aquatic weeds. A U.S. patent was issued on ARS's process for producing and stabilizing blastospores of the bioinsecticidal fungus Paecilomyces fumosoroseus (U.S. Patent #08,968,808 "Methods and Compositions for Producing Desiccation Tolerant Paecilomyces fumosoroseus Spores"). A CRADA partner is pursuing commercial development of this technology and has applied for an exclusive license on the patent.
Foreign patent rights for this process are under prosecution in Europe, Canada, and Mexico. It is envisioned that P. fumosoroseus-based biopesticides may be available as commercial products within 3-5 years. Results of our successful discovery and field testing of biocontrol microbes effective against head scab of wheat have been presented at forums and scientific meetings attended by wheat society members, farmers, agricultural companies, and scientists. These include the National Fusarium Head Blight Forum, the National Association of Wheat Growers, and the Annual Meeting of the American Phytopathological Society.
8. List your most important publications in the popular press (no abstracts) and presentations to non-scientific organizations and articles written about your work (NOTE: this does not replace your peer-reviewed publications which are listed below)
Impacts
(N/A)
Publications
- Jackson, M.A. Microbial biopesticides. Lederberg, J., editor. Academic Press, San Diego, CA. Encyclopedia of Microbiology, 2nd Edition. 2000. p. 541-555.
- Cliquet, S., Jackson, M.A. Influence of culture conditions on production and freeze-drying tolerance of Paecilomyces fumosoroseus blastospores. Journal of Industrial Microbiology and Biotechnology. 1999. v. 23. p. 97-102.
- Vega, F.E., Jackson, M.A., McGuire, M.R. Germination of conidia and blastospores of Paecilomyces fumosoroseus on the cuticle of the silverleaf whitefly, Bemisia argentifolii. Mycopathologia. 1999. v. 147. p. 33-35.
- Connick, W.J., Jr., Ingber, B.F., Goynes, W.R., Jr., Daigle, D.J., Jackson, M.A., Lewis, J.A. American Laboratory. 1999. v. 31. p. 13-16.
- Vega, F.E., Mercadier, G., Guermache, F., Crespy, N., Quimby, P.C., Jackson, M.A. Effect of C:N ratio and nitrogen content on spore yield ... with the "Stabileze" process. Virginia Tech, Blacksburg, VA. Technical Bulletin "Development of Biopesticides for Grasshopper and Locust Control in Sub-Saharan Africa," USAID/Africa Bureau Grant No. AOT-G-00-97-00386-00. 1999. p. 26-38.
- Jackson, M.A. Selecting and optimizing nitrogen sources for fermentation processes. Annual Meeting of the Society for Industrial Microbiology. 2000. Abstract p. 66.
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Progress 01/01/99 to 09/30/99
Outputs
1. What major problem or issue is being resolved and how are you resolving it? Commercial interest in using living microbial agents for controlling agricultural pests such as weeds, insects, and plant pathogens or as probiotic feed additives for livestock is driven by an increasing public demand for environmentally-friendly, sustainable agricultural practices. The widespread use of chemical pesticides and antibiotic feed additives are often viewed as non-sustainable agricultural practices. An integrated pest management (IPM) approach using biological control agents (biopesticides) in conjunction with chemical pesticides is recognized as a sustainable pest management system which reduces chemical pesticide usage and deters the development of pest resistence to the chemical control. Likewise, the use of living microorganisms as feed additives (probiotics) has been shown to improve animal health, weight gain, and feed conversion efficiency. Effective probiotic feed
additives can reduce or eliminate the use of antibiotics in feeds. A significant constraint to the commercial use of living microbial agents in agriculture is the lack of low-cost methods for producing stable, effective microbial products. While living microbial agents can be produced using solid-substrate or liquid culture production methods, liquid culture fermentation processes are generally the most cost-effective. Optimization of deep-tank, liquid fermentation processes for the production of living microbial agents must take into account not only product yield but also microbial stability and product efficacy. Our three major research goals will be to 1) develop low-cost, deep-tank fermentation methods for the production of stable, effective living microbial propagules for use as biocontrol agents or probiotic feed additives, 2) develop methods for drying and formulating fungal and bacterial microbial biopesticides which enhance the viability and stability of these agents
during storage, and 3) identify potential bacterial or fungal biocontrol agents for the control of Fusarium graminearum, the causative agent of head scab of wheat. 2. How serious is the problem? Why does it matter? Public concerns over the widespread use of chemical pesticides and over increases in pest resistence to these chemical controls have heightened interest in developing environmentally friendly pest control measures. The use of microbial pathogens as biopesticides for controlling weeds, insects, and plant diseases is an acceptable alternative control measure. Needs for microbial products that control plant pests remain high. For instance, viable control measures are currently unavailable for Fusarium head blight of wheat and barley which is estimated to cause more than 3 billion dollars in losses annually. Plant and insect pathologists have identified hundreds of microbial agents which show good potential for use as biopesticides, yet, over the past 20 years, less than 15
living microbial biopesticides have been commercially developed. In large part, the limited success in commercializing these microbial agents is due to a lack of economical production methods, poor product shelf-life, and inconsistent pest control under field conditions. Unless these problems are addressed, the widespread use of biopesticides will not become a reality. Research designed to understand how the nutritional and environmental conditions present during the production process affect biopesticide yield, stability, and biocontrol efficacy is needed if cost-effective production methods are to be developed. Likewise, the use of antibiotic feed additives has been implicated in hastening the development of antibiotic-resistant bacteria thus mitigating the usefulness of various classes of antibiotics for human and animal health uses. Many countries in Europe have already banned the prophylactic use of antibiotics in animal feeds. In many cases, living microbial feed additives
have been shown to provide health and weight gain benefits comparable to those seen with antibiotic supplementation. Since the consistent efficacy of these products is dependant on the delivery of a living microbial feed additive, many of the technical constraints applicable to biopesticides are also affecting the commercial availability of probiotics. The widespread use of probiotic feed additives will require the development of low-cost production methods which yield stable, effective microbial feed additives. 3. How does it relate to the National Program(s) and National Component(s) to which it has been assigned? This research focuses on the development of cost effective fermentation processes for mass producing microbial biopesticides which contributes to National Program 304 "Crop and Commodity Pest Biology, Control and Quarantine" (70%). The expected outcome from this research is the commercial use of microbial biopesticides and probiotic feed additives which will lead to
additional markets for the agricultural commodities used as nutrients in the fermentation process thus contributing to National Program 306 "New Uses, Quality and Marketability of Plant and Animal Products" (30%). In addition, the use of biopesticides will reduce pest damage to plant products, thereby improving product quality and marketability. The research directed at discovering and developing microorganisms for biologically controlling head scab of wheat demonstrates a strong association with the Biological Control component of National Program 303 "Plant Disease." 4. What were the most significant accomplishments this past year? In order to use portable fermentation equipment for the onsite production and delivery of living fungal biocontrol agents such as spores of Paecilomyces fumosoroseus, process conditions had to be developed which favor the rapid production of P. fumosoroseus spores while inhibiting the growth of undesirable microbes. Nutritional and environmental
conditions were optimized for the liquid culture production of spores of P. fumosoroseus using an aseptic, non-sterile portable fermentation system. These studies showed that high concentrations of P. fumosoroseus spores could be rapidly produced in portable fermentors with little or no bacterial contamination when the pH of the production medium was lowered and appropriate nutrient concentrations were provided in the production medium. These studies have demonstrated that the onsite production of fungal biocontrol agents is feasible and, specifically, that high concentrations of P. fumosoroseus can be rapidly produced using portable, aseptic fermentation equipment. Farm losses due to scab of wheat have been conservatively estimated to exceed 3 billion dollars in the 1990's in the United States alone with devastating losses occurring in the Upper Midwest states, Illinois, Indiana, and Ohio. In laboratory and field studies conducted at the NCAUR in Peoria, Illinois, and field
studies conducted at Wooster, Ohio, a USDA-ARS research plant pathologist and Ohio State University scientists demonstrated that microbes they discovered greatly reduced the severity of scab of wheat, scab disease incidence, and mycotoxin content of wheat. The most effective microbes reduced disease severity in field trials by as much as 75% with disease reduction being demonstrated on three different types of wheat. With this discovery, the feasibility of biologically controlling scab of wheat has been demonstrated, and a crucial step towards the development of a biological control product for use against this disease has been completed. 5. Describe the major accomplishments over the life of the project including their predicted or actual impact. While this CRIS project is in its first year, it builds on our previous CRIS project #3620-41000-057. Our previous work led to the development of deep-tank fermentation methods for the production of three promising microbial
biopesticides. The nutritional composition of these liquid culture production media was optimized to maximize yields, propagule stability, and biocontrol efficacy. This novel research showed that the diets fed microbes during the production process could have a profound influence on the quantity and quality of the biopesticidal product. The results and impact of our work under CRIS #3620-41000-057 are presented below. Nutritional studies led to the development of a unique liquid culture medium which produced high yields of microsclerotia (compact, highly resistant cell masses formed by many fungi as overwintering structures) of the bioherbicidal fungus Colletotrichum truncatum. This was the first report of a liquid medium for the deep-tank production of fungal microsclerotia (MS). When these MS of C. truncatum are incorporated into soil, they infect and kill emerging seedlings of the weed hemp sesbania. Under refrigerated conditions, over 90% of the MS survived 4 years' storage.
The development of a liquid culture production method, coupled with the stability and biocontrol efficacy of MS of C. truncatum, has piqued commercial interest in their use as a biopesticide. These studies demonstrated that the regulation of nutritional conditions during culture growth can yield novel cellular structures which may provide alternative approaches to biocontrol. A liquid culture medium was developed which yielded high concentrations of the bioherbicidal bacterium Xanthomonas campestris. This bacterium selectively infects and kills annual bluegrass, a weedy grass in various turfgrass systems. Cells of X. campestris harvested after the cultures depleted essential nutrients from the medium and entered a "stationary" growth phase survived drying and storage better than cells harvested earlier in growth during the exponential phase of growth. These studies demonstrated that stable cellular preparations of X. campestris could be produced in deep-tank fermentation and
stored as a dry preparation for over six months with little loss in viability. This product is currently under commercial development in the United States and is a commercial product in Japan for the control of annual bluegrass in turf. Our nutritional studies with the bioinsecticidal fungus Paecilomyces fumosoroseus (Pfr) culminated in the development of a patented liquid culture medium which supports the deep-tank production of high concentrations of stable Pfr spores. This was the first report of a liquid culture medium which supports the production of desiccation-tolerant spores of this fungus. Collaborations with ARS insect pathologists have shown that these liquid culture produced Pfr spores are highly infective on the silverleaf whitefly, a major insect pest in greenhouses and in arid climates, worldwide. Media optimization studies led to the development of a rapid (2 day) Pfr spore production process which yields very high spore concentrations (1 x 10**9 spores/ml). It is
expected that our Pfr spore production process will become a commercial method for producing this promising bioinsecticide. In collaboration with industry, the commercial potential of this production process is currently being evaluated. Under our new CRIS project, work has continued on the optimization of production and stabilization processes for P. fumosoroseus spores. In order to use portable fermentation equipment for the onsite production and delivery of fungal biocontrol agents such as spores of P. fumosoroseus, process conditions were developed which favor the rapid production of P. fumosoroseus spores while inhibiting the growth of undesirable microbes. High concentrations of P. fumosoroseus spores were rapidly produced in a portable fermentor with little or no bacterial contamination when the pH of the production medium was lowered and appropriate nutrient concentrations were provided in the production medium. These studies have demonstrated that the on-site production of
fungal biocontrol agents is feasible and, specifically, that high concentrations of P. fumosoroseus can be rapidly produced using an aseptic, portable fermentation system. In laboratory and field studies conducted at the NCAUR in Peoria, Illinois, and field studies conducted at Wooster, Ohio, biocontrol microbes were isolated and tested which greatly reduced the severity of scab of wheat, scab disease incidence, and mycotoxin content of wheat. The most effective microbes reduced disease severity in field trials by as much as 75% with disease reduction being demonstrated on three different types of wheat. With this discovery, the feasibility of biologically controlling scab of wheat has been demonstrated and a crucial step towards the development of a biological control product for use against this disease has been completed. 6. What do you expect to accomplish, year by year, over the next 3 years? During FY 2000, field trials will be conducted to control the silverleaf whitefly in
vegetable crops and greenhouses using spores of P. fumosoroseus (Pfr) which have been produced and dried in our laboratory and which have been produced onsite using portable fermentors. Field trials designed to evaluate the efficacy of Pfr spores produced in onsite fermentors for silverleaf whitefly control in melons will be conducted this fall in collaboration with scientists at the USDA-ARS Subtropical Agricultural Research Laboratory in Weslaco, Texas. Additional field trials using this onsite Pfr production system are planned for whitefly or aphid control in greenhouses. Formulation and stabilization studies with Pfr spores will continue with the goal of increasing the shelf-life of this bioinsecticidal agent. In addition, studies will be initiated to discover formulations that enhance the effectiveness of biocontrol agents which have been shown to reduce or eliminate head scab of wheat. Initial studies will identify carbon and nitrogen containing compounds that can be
utilized by scab antagonists but not by the wheat scab pathogen, Fusarium graminearum. These compounds will then be used to formulate scab antagonists in greenhouse trials with the likely result of increasing the population and disease control activity of our antagonists after they are placed on wheat heads without benefitting the growth of the pathogen. We will also likely demonstrate, for a third year, field test effectiveness of microbial strains identified as being suppressive to scab of wheat in greenhouse and field experiments. In addition to field studies being conducted at Peoria, Illinois, and Wooster, Ohio, cooperative field research with North Dakota State researchers is being planned. During FY 2001, field trials will be conducted in collaboration with an industrial partner to optimize onsite Pfr spore production and to evaluate Pfr efficacy against various insect pests in differing agricultural settings. Successful field trials using onsite-produced Pfr spores will be
followed by experiments required to obtain EPA registration of onsite produced Pfr spores for control of whiteflies and other soft-bodied insects. It is envisioned that the commercialization of this bioinsecticide will provide farmers and greenhouse operators with a cost-effective pest management tool. New studies will be initiated in FY 2001 to optimize mass production methods for living microbial feed additives. GRAS microbes such as lactobacilli or yeasts are likely candidates for these studies. Also during FY 2001, ARS will, in cooperation with partners, field test those formulations found to enhance the effectiveness of biocontrol agents of scab of wheat. Studies will also be continued to develop nutritional and chemical environments for antagonist production via liquid culture so as to optimize the quantity, effectiveness, and shelf life of cells of biocontrol agents produced. In association with fermentation studies, molecular biology approaches will be utilized to
determine the mechanism(s) utilized by successful microbes in reducing the severity of scab disease. During FY 2002, ARS will, in cooperation with partners, field test cells of biocontrol agents that were produced using the optimal liquid culture conditions discovered during FY 2001 research. These optimally produced biocontrol agents could then also be field tested in combination with the best resistant varieties and small quantities of the best available fungicides to determine if these integrated disease management tools can be used to provide more effective control of scab disease. 7. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end user (industry, farmer, other scientists)? What are the constraints if known, to the adoption & durability of the technology product? The data obtained from these studies have been presented at national and international meetings and have been published in
peer reviewed journals. These data provide other researchers in this area of research with valuable information on how nutritional regulation of culture medium can be used to maximize the yield of stable effective biopesticidal propagules. Using our media optimization strategies for developing liquid production media for biopesticides, we are collaborating with scientists in France, Spain, Japan, and Mexico on developing production media for various biopesticides. This work has led to a Specific Cooperative Agreement between ARS and academic scientists in Mexico to develop low-cost commercial methods for producing and formulating fungal biopesticides. A U.S. patent has been approved on our process for producing and stabilizing blastospores of the bioinsecticidal fungus Paecilomyces fumosoroseus (U.S. Patent #08/944,245 "Methods and Compositions for Producing Desiccation Tolerant Paecilomyces fumosoroseus Spores"). A Cooperative Research & Development Agreement (CRADA) partner is
persuing commercial development of this technology and has applied for an exclusive license on the patent. Foreign patent rights for this process are under prosecution in Europe, Canada, and Mexico. It is envisioned that a P. fumosoroseus-based biopesticides may be available as commercial products within 3-5 years. We have presented the results of our successful discovery and field testing of biocontrol microbes effective against head scab of wheat at forums and scientific meetings attended by wheat society members, farmers, agricultural companies, and scientists. These include the ARS Fusarium Workshop in Athens, Georgia; the Fusarium Scab and Toxins in Wheat and Barley Workshop, Peoria, Illinois; the National Fusarium Head Blight Forum, E. Lansing, Michigan; Annual Meetings of the American Phytopathological Society and the American Society of Microbiology; and a meeting of the Illinois Wheat Association, Springfield, Illinois. The application of this technology towards controlling
scab on durum wheats is currently being tested and the results will be conveyed to a growers cooperative for durum wheats. The timing of when this technology becomes available to the end user is dependent on attracting a company in the agricultural industry that could cooperatively bring this technology to the marketplace. Should pending patents be granted and a suitable industrial partner be identified, commercial scale field testing could begin as early as the 2001 growing season. The technology is already available to other scientists via the publication of our research results in scientific journals. Constraints to the adoption of this technology would be few given that alternative means of controlling this disease are currently not available. Biological control is generally considered to be a durable method for controlling plant diseases since pathogens generally are not successful in developing resistance to microbes that biologically control plant disease. 8. List your
most important non-peer reviewed publications and presentations to non-scientific organizations, and articles written about your work(NOTE: this does not replace your peer reviewed publications which are listed below). "Scientists Finding Answer to Crop Infection," Journal Star Newspaper, Peoria, IL, November 29, 1998. "Dakota Growers Teams with Researcher to Develop Natural Scab Fighters," North Dakota Agribeat, John MacDonald, Associated Press, April 4, 1999. "Deep-Tank Fermentation - for Cheaper Bioinsecticides," Agricultural Research, January 1999, p. 25. "Model Bioinsecticide Aimed at High-Value Crops," Illinois Agrinews, January 15, 1999, p. D4. "Portable Fermentors Make Bioinsecticide at Site," Industrial Bioprocessing, February 1999, 21:8-9. "Fungus Promises Weapon Against Silverleaf Whitefly," Florida Grower, February 1999, p. 12. "Whitefly Bio Control - Bioinsecticides are the Wave of the Future," American Vegetable Grower, March 1999, p. 2-3. "Fighting Fusarium,"
Agricultural Research, USDA-ARS, June 1999, pp. 18-21.
Impacts
(N/A)
Publications
- JACKSON, M.A. 1999. Methods and compositions for producing desiccation tolerant Paecilomyces fumosoroseus spores. U.S. Patent #08/944,245, Allowed February 19, 1999.
- POPRAWSKI, T.J. and JACKSON, M.A. 1999. Laboratory activity of blastospores of Paecilomyces fumosoroseus on Bemisia argentifolii nymphs, 1997. Arthropod Manage. Tests 24:399-400.
- JACKSON, M.A., FRYMIER, J.S., WILKINSON, B.J., ZORNER, P. and EVANS, S. 1998. Growth requirements for the production of stable cells ... Xanthomonas campestris. J. Ind. Microbiol. 21:237-241.
- VIDAL, C., FARQUES, J., LACEY, L.A. and JACKSON, M.A. 1998. Effect of various liquid culture media on morphology, growth, ... Paecilomyces fumosoroseus. Mycopathologia 143:33-46.
- HERNANDEZ-TORRES, I., GALAN-WONG, L., JACKSON, M.A. and PEREYRA-ALFEREZ, B. 1999. A Paecilomyces fumosoroseus ... Bemisia tabaci. Annu. Meet. Soc. Invertebr. Pathol. Abstr. p. 44.
- JACKSON, M.A., VEGA, F.E. and POPRAWSKI, T.J. 1999. Optimized conditions for the production ... fungus Paecilomyces fumosoroseus. 99th Annu. Meet. Am. Soc. Microbiol. Abstr. p. 574.
- VEGA, F.E., MERCADIER, G. and JACKSON, M.A. 1999. Insect pathology at USDA's European biological control laboratory. Annu. Meet. Soc. Invertebr. Pathol. Abstr. p. 77.
- VEGA, F.E., MERCADIER, G. and JACKSON, M.A. 1999. Fungi associated with the coffee berry borer, Hypothenemus hampei. 7th Eur. Meet. IOBC/WPRS Abstr. p. 14.
- VANDENBERG, J.D., SANDVOL, L.E., JARONSKI, S.T., JACKSON, M.A., ... HALBERT, S.E. 1998. Field use of ... Diuaphis noxia. VII Int. Colloq. Invertebr. Pathol. Microb. Control Abstr. p. 23.
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