EPA Workplan: Demonstration Project on Hydrilla and Hygrophila in the Upper Kissimmee Chain of Lakes

Element 3: Biological Control of Hydrilla and Hygrophila

Objectives Addressed by Element 3

1) To evaluate the effectiveness of biological controls in the treatment of hydrilla and hygrophila.

2) To evaluate new technology processes or practices, or a new combination or uses of technologies, processes or practices for the control of hydrilla and hygrophila using small-scale field work.

3) To implement and monitor successful practices and processes identified in objectives 1 and 2 using large-scale field demonstrations.


Classical insect biological control can be particularly successful at controlling invasive pests of foreign origin. The underlying principal of classical biological control is that pests are kept in equilibrium in their native ranges by a complex of biological checks and balances such as insects, diseases, nematodes and other biological organisms. When introduced into another geographical location, away from these natural checks, the organism proliferates and becomes a serious pest.

Thus, classical biocontrol of weeds consists of determining the native range of an exotic plant and searching that range for natural checks and balances with the goal of determining safety of the controlling organism and introducing it into the area where the weed is a pest.

The search for biocontrol insects for hydrilla has concentrated on seeking herbivores of hydrilla in East and Southeast Asia and Australia. Several insects have been found that feed only on hydrilla and have been introduced into the US. These insects have not however been predictable and have not significantly reduced the need for hydrilla management by other means.

There have been no attempts, surveys or research conducted on the potential for insect biocontrol of hygrophila. This has been due to the limited range of this Asian species in the US; i.e. it has not become a serious, widespread pest to attain sufficient attention for the relatively high cost of insect biocontrol research, despite the fact that scientists feel that there is good potential for discovery of successful biocontrol insects.

The use of plant pathogens, or diseases of plants caused by bacteria, fungi and other micro-organisms, or chemicals produced by these microorganisms, has been successful for some terrestrial weeds. The objective of the use of mycoherbicides for weed control is to culture appropriate natural microorganisms and spray the weeds with rates high enough to overwhelm the plants ability to recover from the induced disease. There are literally hundreds if not thousands of potential mycoherbicides found in nature, so scientists generally chose candidates from those groups of microorganisms that have historically been effective on other weeds.

One potential pathogen of hydrilla, Micoleptidiscus terrestris or Mt, was discovered on submersed weeds in the 1970's and has been evaluated periodically since that time. The problem with Mt has been the inability to develop cultures with high enough concentrations of Mt to be effective in the highly variable aquatic environment. Preliminary testing has also shown that some members of the Actinomycetes family has activity against hydrilla. None of these potential mycoherbicides have been developed in part due to the lack of commercial interest.


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Element 3, Task 1: Demonstration of Hydrilla Control in Osceola County, Florida using Mycoleptodiscus terrestris (Mt), a New Contact Bioherbicide, for Aquatic Plant Management (Project ongoing)


1) To evaluate the effectiveness of biological controls in the treatment of hydrilla.

2) To evaluate new technology processes or practices, or a new combination or uses of technologies, processes or practices for the control of hydrilla using small-scale field work.

3) To implement and monitor successful practices and processes identified in objectives 1 and 2 using large-scale field demonstrations.


First discovered in the early 1970’s, Mycoleptodiscus terrestris still remains to be fully developed as an operational tool for aquatic plant management. Early developmental efforts failed to reach the level of product stability and virulence needed to allow the aquatic plant pathogen to be fully commercialized. Starting in 2001, a cooperative research effort was initiated between the USDA-ARS-NCAUR in Peoria, IL (PI: Dr. Mark Jackson), the Aquatic Plant Control Research Program at the Environmental Laboratory of the US Army Engineer Research and Development Center (ERDC) in Vicksburg, MS (PI: Dr. Judy Shearer), and SePRO Corporation headquartered in Carmel, IN with research facilities in Whitakers, NC (PI: Dr. Mark Heilman). This ongoing collaborative effort has developed new fermentation methods that enhance Mt virulence on hydrilla while providing a new dry, stable Mt material that can be incorporated with other materials and stored for several months or more prior to use. SePRO has negotiated a licensing agreement with USDA and the US Army Corps of Engineers to commercialize new aquatic plant management technologies resulting from these new, patented fermentation methods for aquatic plant pathogens, including Mt. Latest research efforts have focused upon development of new dry Mt formulations using the new production methods with the goals of enhancing attachment of Mt propagules to treated hydrilla and further improving overall performance to reach commercially viable levels of activity and efficacy. History of biocontrol using plant pathogens has shown that formulation adjustments can have profound impact on the effectiveness of such pathogens for plant pest control. Early work has documented a several fold increase in Mt activity on hydrilla by addition of relatively simple formulating agents to the new dry formulation. Additional work by Dr. Shearer, Dr. Linda Nelson, and others at the ERDC has also documented strong interaction between Mt and other aquatic herbicides for hydrilla control.

By early 2007, newly formulated, dry Mt materials should be ready for field evaluation. The scope of work presented here reviews a four-year plan for field demonstration in Osceola County conducted in conjunction with other field evaluation activities and final development of a new, commercially available, biocontrol alternative for hydrilla management. USDA, ERDC, and SePRO will conduct activities through continued public/private research collaboration. Along with internal funding by these government and industry research partners, all Osceola County support for this program is anticipated to be combined with additional research funding by the Bureau of Invasive Plant Management of the Florida Department of Environmental Protection (BIPM) and the Biopesticide section of IR-4 to accelerate field-testing and final development of the Mt aquatic herbicide.


Field-testing and evaluation is needed in two primary areas to finalize Mt commercially available for management of hydrilla in Osceola County and in other affected areas of Florida and the United States. Developmental activities are described for a four-year period beginning October 2006. It should be noted that funded activities are assumed to be subject to annual renewal based on review by funding agencies confirming appropriate level of progress.

  • Production and evaluation of new formulations of dry Mt product. Since 2004, SePRO and its government collaborators have focused on refining pilot-plant fermentation and production protocols and have developed initial prototypes of new formulations of Mt material. Such work to date has been limited by availability of equipment needed for full pilot-plant capability at USDA in Peoria and insufficient human resources able to be devoted for rapid lab and field evaluation of new materials. Support from EPA/Osceola County and FLDEP-BIPM will assist final equipment acquisition and also support hiring of additional staff at USDA, ERDC, and SePRO to increase effort available to examine performance of final formulations of Mt herbicide. This evaluation effort will include:
  • Intensive, collaborative laboratory activities by USDA, ERDC, and SePRO focused on manipulating production materials and methods, and efficacy testing of new formulations on hydrilla. Effort will be made to develop improved, more efficient methods to confirm efficacy of final Mt formulations under flask or similar, smaller-scale test conditions prior to field use.
  • Outdoor evaluation at the SePRO Research and Technology Campus (RTC) in Whitakers, NC and at the US Army Corps Lewisville Aquatic Ecosystem Research Facility (LAERF) in Lewisville, TX. Outdoor testing will begin in small tanks and then move to larger mesocosm tanks available at the LAERF. Most effective formulations will be evaluated in partial or full pond trials at the LAERF and the RTC. Both the LAERF and RTC are already permitted to conduct such trials on their properties.
  • Field Demonstration in Osceola County, FL. The most effective Mt formulation(s) will be produced under internal or commercial pilot-plant conditions to allow Experimental Use of Mt herbicide for treatment of between 1 – 50 acres of hydrilla in pond or lake sites located in Osceola County, Florida. The scale of treatments will be determined by the developmental status of the product and the availability of appropriate sites within Osceola County. The scale of field demonstration is projected to increase through the lifetime of the work plan. The demonstration phase will increase in scope during the program and include production, application, and evaluation of full commercial-scale fermentations of Mt (typical commercial batch – 30,000 gal fermentation volume).
  • Evaluation of herbicide interaction between Mt formulations and existing/new chemistries for hydrilla management. A parallel effort of evaluation of such interactive effects will occur simultaneously with the final evaluations of the best Mt formulation. By lowering use rates of both pathogen and traditional herbicide like fluridone or endothall, Mt performance may reach levels of commercial viability sooner in the development process while enhancing the efficacy of the existing herbicide. Several new herbicides for hydrilla management are also under development, and interaction of Mt with those technologies will also be examined as they become available.
  • In conjunction with field demonstration and final development activities described above, laboratory evaluations will occur at the ERDC in Vicksburg and at the SePRO RTC in NC. Shearer and Nelson have conducted such evaluations for several existing herbicides in the past, but further testing will be initiated as final formulations of Mt are trialed in the field.
  • The focus of outdoor, controlled interaction testing will be outdoor mesocosm tanks at LAERF. These systems are specifically designed to allow efficient manipulation of contact time and are large enough to allow effective examination of interactive effects on not just hydrilla, but also a suite of other potential target and non-target plants in the same trial. One benefit of Mt-herbicide interaction may be improved treatment selectivity, and this can be effectively examined under such test conditions.
  • Field Demonstration in Osceola County, FL. A minimum of three test sites selected for field evaluation in Osceola County will be simultaneously treated with Mt and other aquatic herbicides for examination of field-scale interaction. Methods evaluated at the mesocosm scale will be confirmed through these combined trials under realistic field conditions.
  • Specifics of Work Plan Using Osceola County Funds from EPA Grant #X796433105-0

Note: Schedule described below assumes October 2006 start date for funding from Osceola County. As per EPA guidelines, a full Quality Assurance (QA) plan will be developed and submitted for final review and authorization prior to initiation of grant-supported data collection.

Production of New Formulations: Since 2004, USDA has been focused on developing best methods of production of dry Mt materials to allow efficient production and evaluation of new dry Mt formulations at a pilot-scale. Most of the equipment needed for such effort is in place with the exception of a $50,000 drying system that would mimic commercial pilot conditions and improve processing of Mt materials for larger scale production and evaluation. An additional formulation chemist (Dr. Chris Dunlap) was hired by USDA in 2004 to help improve capability in formulation development. SePRO has also recently added a formulation chemist who, through support from FLDEP and EPA/Osceola, will spend 15-20% of his time on this project to help support USDA internal efforts. While efforts with new materials are showing great promise, continued work is needed. Additional lab technical support is needed to help aggressively produce and evaluate the germination, sporulation, and overall viability of new dry Mt materials once formulated. Increased focus will be given to production and characterization of best Mt dry formulations in conjunction with ERDC and SePRO determination of performance on hydrilla. With additional technician support to USDA through this proposed funding, new batches of formulated product should be able to be produced on approximately a two-week schedule, and production scale increased to support field study needs. After best Mt formulation is selected for full commercial production based on efficacy testing in the laboratory and field, USDA and SePRO will work with a commercial fermentor contracted by SePRO to outline the conditions of fermentation and production needed to successfully produce the new formulation. With Osceola and FLDEP support, the first commercial pilot runs would be projected for 2007, and progressively larger runs would be conducted in 2008 – 2010 with potential to accelerate commercial production based on results of the program. All formulated Mt product for field demonstration produced in conjunction with Osceola County and BIPM support would be produced at cost for this project.

Laboratory Testing of New Formulations: In order to fully evaluate new Mt formulations as quickly as possible, Drs Shearer and Heilman will oversee the conduct of monthly aquarium-scale trials at both the ERDC in MS and the SePRO RTC in NC for the duration of the project (2006-2010). The combined effort of both laboratories should be able to effectively evaluate new batches of formulated Mt that would be developed on roughly a two-week schedule by Dr. Jackson’s laboratory in Peoria. Each aquarium trial will consist of a minimum of 30 test units (10 herbicide rates, 3 replicates per rate). Approximately 20% of aquarium efforts will involve simultaneous treatments with other herbicides to look at interaction with Mt. Direct funding in 2006 would also support ERDC (Shearer and Nelson) to evaluate new methodology for small-scale testing of Mt. In addition, from 2006 through 2008 in a 1:2 sharing between Osceola County and SePRO, the collaborative research team will fund a Masters-level graduate student working out of the laboratory of Dr. Rob Richardson at North Carolina State University near SePRO’s RTC facility in NC. This student and Dr. Richardson will collaborate with USDA, ERDC, and SePRO to help develop improved laboratory-scale methods for predictions of Mt field performance and investigate ecophysiology of Mt-hydrilla interaction. The student will have the benefit of direct access to SePRO facilities and opportunity to compare his/her results with other lab, aquarium and outdoor testing occurring at the RTC.

Outdoor Evaluation of New Formulations: Outdoor testing in small tanks to confirm efficacy will occur in greenhouse or shadehouse at the RTC. A minimum of 6 outdoor small tank trials (approx. 30 reps; 10 rates with 3 reps per rate) will be performed annually during each year of the project with most occurring between April 1 and October 1 each year. In addition, mesocosm-scale trials through ERDC (Nelson and Shearer) at the LAERF will be conducted in the first two years of the project. These trials will use 30-2,000-gal tanks available at the LAERF. Each study will examine effects of Mt application on hydrilla and approximately 4 native aquatic plant species. Such species will vary between studies but would likely include a selection from key Florida native submersed and floating-leaved plants such as Ceratophyllum demersum, Potamogeton illinoensis, Vallisneria americana, Nuphar advena, and Nymphaea odorata. One mesocosm study at LAERF under FLDEP funding would occur in 2006. One trial at LAERF would also be conducted in 2007 and two during the spring/summer of 2008. Studies would focus on efficacy and selectivity of Mt alone and in conjunction with other herbicides, building on knowledge from each previous study to refine knowledge of best Mt and other herbicides rates for field demonstration protocols. Trials would not be conducted at LAERF in 2009 or 2010, as large-scale demonstration studies in Osceola County would become primary focus of final field development effort. Some additional confirmation testing would also be conducted at University of Florida in all years of the project.

Initial fieldwork in Osceola County would begin in the spring of 2007. SePRO will work with Osceola County in 2006 to identify appropriate test sites for 2007 and later years of the project. An Experimental Use Permit will be needed to commence this work, and SePRO will initiate discussions with EPA in 2 nd half of 2006 to acquire this permit for 2007 - 2010. Studies in early 2007 would compare efficacy in small, curtained plots (25 – 100 sq ft) within ponds in Osceola County. Follow-up work in later 2007 would test efficacy of Mt from first commercial production in larger closed or open plots up to 1 acre in size. In 2008, Mt material produced under commercial pilot-scale conditions would be evaluated in treatments totaling between 5 – 50 acres depending on test site characteristics and product best use rate at that stage of development. Assuming positive results of these tests, SePRO would conduct any necessary registration studies and submit for Section 3 registration by late 2008. In 2009 and 2010, full-scale commercial fermentations would be produced to allow treatment of between 100-200 acres in both years. Protocols would focus on field treatments between 5 – 50 acres each in size. In all years of the project, some study sites would be selected based on opportunity to study interaction with treatments of other complementary herbicide modes of action based on previous interaction testing. Field effects on target hydrilla and any non-target aquatic plant species will be evaluated with an emphasis on duration of control with or without other herbicide use. All field evaluations will be tied to pre-treatment assays of Mt material developed as a part of this program and post-treatment assays of field infection of target and non-target plants that have been utilized by ERDC in the past to confirm Mt activity. The University of Florida would be contracted to assist in field evaluations of hydrilla and other vegetation impacts.

Results of all field demonstrations within Osceola County will be presented in annual, public, progress reports and described as appropriate in the context of all public and private research efforts to introduce Mt as a bioherbicide for hydrilla management. Report format and dissemination will be tailored to needs of Osceola County and EPA.

Schedule of Activities for Element 3, Task 1:


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Element 3, Task 1 Milestones

Year 1:

  • Development and implementation of EPA QA plan for all laboratory and field demonstration studies. Proposed QA plan will be submitted in April 2006, phased into ongoing USDA/ERDC/SePRO collaborative research in summer 2006 prior to full implementation upon initiation of Osceola County/ EPA project support.
  • Laboratory and outdoor evaluations of new Mt formulations (USDA, RTC, ERDC)
  • Initiation of collaborative research with NC State to develop improved, predictive laboratory methods for evaluating new formulations and study ecophysiology of Mt-hydrilla interaction (4 th quarter)
  • Identification of potential field sites in Osceola County for 2007 (and add’l years) and submission of EUP request to treat sites. Small plot field demonstrations in Osceola County, FL under EUP: 1 – 10 total acres projected for treatment
  • First commercial pilot-scale production of Mt
  • Outdoor testing of best formulations at RTC and LAERF
  • Small plot field demonstrations in Osceola County, FL under EUP: 1 – 10 total acres projected for treatment

Year 2:

  • Continued laboratory evaluations of Mt formulations and new methods
  • Outdoor testing at RTC and LAERF
  • Expanded commercial pilot-scale production of Mt
  • Expanded field demonstrations in Osceola County, FL under EUP: 5 – 50 total acres treated

Year 3:

  • Registration Studies and Submission of Section 3 Registration .
  • Continued laboratory evaluations of Mt formulations and new methods
  • First full-scale commercial production of Mt
  • Large operational-scale testing in Osceola County, FL under EUP (or registration if available): 100 – 200 total acres treated with some combination with other herbicides

Year 4:

  • Continued laboratory evaluations of Mt formulations and new methods
  • Additional full-scale commercial production of Mt
  • Large operational-scale testing in Osceola County, FL under registration: 100 – 200 total acres treated with emphasis on combinations with other herbicides


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Element 3, Task 1 Measure of Success

Full scale commercial production of Mt accomplished and large scale testing in Osceola County completed.

Proposed Collaborative Budget for Element 3, Task1:

For the final development and field demonstration of Mt as an effective bioherbicide for hydrilla control, total fiscal support for the studies is projected to come from six sources: USDA, ERDC, Osceola County through EPA, FLDEP BIPM, IR-4, and SePRO Corporation. The table on the following page describes annual and total expenditures anticipated by each party for various categories to support the above scope of work during the multiple-year period of the project. Additional information is available upon request. The grand total projected contribution of Osceola County through EPA grant over the lifetime of the project would be $500,000. Final procurement of financial support from the BIPM and IR-4 is pending. BIPM feedback concerning support for this project in conjunction with EPA/Osceola grant has been very favorable.

At the start of every project year, it is anticipated that a full review of project status will occur and a decision made on further evaluation plans based on annual progress, overall project status, and grant funding priorities.


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Element 3, Task 2: Foreign Exploration for Natural Enemies of Hygrophila

Objectives addressed in Element 3

1) To establish collaboration with scientists and institutes in the native range of hygrophila, seek natural enemies of hygrophila, and culture and evaluate these for potential introduction into quarantine in Florida.


Hygrophila polysperma or East Indian Hygro, native to India, was introduced into Florida as an aquarium plant in the 1950's and has slowly spread throughout the state, north to Virginia. Continued expansion to the north and west of Florida is expected. Hygrophila usually occurs in flowing systems, canals, creeks and rivers and is particularly difficult and expensive to control with aquatic herbicides. There have been no studies of significance that have been conducted to seek biocontrol agents to date. This project is the backbone initial steps that need to be taken to find a bio-control agent and future funding will be needed for the continued evaluation of a hygrophila bio-control agent.


Institutions and scientists in the native home of hygrophila will be sought as cooperators in this study. Populations of hygrophila in India will be located by consultation with taxonomists and reviewing herbaria records. Natural populations of hygrophila will be sampled for insect herbivores by standard insect collection techniques, sweep nets, black lights and maintaining in culture to allow any insect eggs and larvae to develop. Insects will be identified to the extent possible and cultured on hygrophila to increase populations and placed back on insect free hygrophila to determine the extent of herbivory. Following preliminary testing for host specificity in India, insects will be considered for seeking USDA approval for introducing into quarantine in the US.

Element 3, Task 2 Milestones

Year 1 will involve travel to India to establish cooperators (being a new project, this establishment can be quite lengthy), search herbaria records and initiate searches on hygrophila in India. Recruitment of a graduate student will also be conducted in Year 1 with this student surveying insects in India. Insects found in India will be evaluated in India for host specificity in Year 2 and 3, and if approved by USDA/APHIS, insects will be imported for further study in the US in quarantine during year 3/4.


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Element 3, Task 3: Collaborative Effort to Search for Natural Enemies of Hydrilla in East Africa (Project ongoing)

Objectives addressed in Element 3

1) To evaluate the effectiveness of biological controls in the treatment of hydrilla.

2) To evaluate new technology processes or practices, or a new combination or uses of technologies, processes or practices for the control of hydrilla using small-scale field work.


The search for natural enemies (insects primarily) of the invasive submersed weed hydrilla has been concentrated in Southeast Asia and Australia over the past 30 years. Several potential insect biocontrol agents have been introduced, but these have not had significant impact on hydrilla growth.

Hydrilla has been present, according to herbaria records, in East Africa for 150 years, but it has never become a problem and occurs in low densities with other native submersed species. There is a distinct possibility that hydrilla is native to East Africa. If in fact it is, the geographical area may have herbivores that are specific to hydrilla and it has never been adequately surveyed for potential insect biocontrol agents.

A preliminary hydrilla insect survey (2 weeks) in Kenya, Uganda and Barundi in Fall of 2005 by UF entomologists Overholt and Cuda located several areas with hydrilla and numerous insects were found in association with the plants. A collaborative proposal has been developed requesting funds jointly from Florida DEP and water management districts to support a concerted 3-4 year effort for seeking and evaluating potential biocontrol insects in East Africa. Not all the agencies will support the project annually, so we are requesting $75,000/yr. from the EPA aquatic weed demonstration project to help fund this project.

Activity 1: Establish collaboration with institutions in East Africa.

A collaborative agreement with International Center of Insect Physiology and Ecology (ICIPE) in Kenya, will be established early in the project. In turn, ICIPE will sign work agreements with the National Institute for the Environment and Conservation of Nature in Burundi and the National Agricultural Research Organization in Uganda.

Activity 1 Milestone

All agreements will be signed between collaborating institutions during Year 1 – FY 2007.

Activity 2: Develop a GIS database and map hydrilla populations in East Africa.

Literature, herbaria and new field survey data will be used to develop a geo-referenced database of hydrilla occurrence in East Africa. During surveys, sites where hydrilla is found will be characterized (water depth, temperature, hydrosoil properties, clarity, etc.). Maps of the hydrilla distribution will be produced from the database, and used to guide exploratory efforts. The database and maps will continually be updated throughout the life of the project as new information from surveys becomes available. Mapping is an important tool in order to locate potential biocontrols.

Activity 2 Milestone

Database and maps produced and posted on the worldwide web. The first maps will be available within the first month of the project.

Activity 3: Molecular characterization of hydrilla.

Hydrilla specimens will be analyzed for molecular structure and compared to Florida hydrilla molecular structure in order ascertain whether the hydrilla is similar enough in structure for biocontrol viability. The samples will be collected from all locations where it is found during surveys in Africa and preserved in 95% ethanol. Hydrilla will also be collected from water bodies in Florida and, through collaborators, from locations where monoecious hydrilla occurs in the US. Efforts will also be made to obtain samples from Asian countries. DNA will be extracted from leaf tissue from all samples. The degree of population genetic relatedness among all sampled locations will be determined by sequencing a variety of regions on the maternally-inherited chloroplast DNA (cpDNA) for which universal primers have already been developed. In addition, 10-15 nuclear, biparentally inherited microsatellite loci specific for hydrilla will be isolated and all individuals will be typed at these loci. CpDNA variation will be useful for describing large scale geographic patterns while the high mutation rate of microsatellite loci results will assist in fine-scale resolution. Using both of these marker types will provide insight into whether hydrilla was introduced into Florida from a single or from multiple source populations, how closely related the source populations are to each other, and the degree of hybridization between source populations. These data can also identify the source region(s) and compare levels of genetic diversity between the introduced and native ranges.

Activity 3 Milestones

Year 1: At least 50 hydrilla samples collected and sent to the University of Miami for analyses.

Years 1-2: DNA extraction methods, cpDNA and microsatellite DNA sequencing techniques established.

Year 3: Comparative studies on cpDNA and microsatellite DNA completed.

Activity 4: Collection of hydrilla insect herbivores.

Hydrilla plants will be collected in Africa from boats and the shore using drag rakes. Plants will be placed in 5 gal. plastic buckets with water and covered with fine cloth netting to allow emergence of endophytic and exophytic insect herbivores. Buckets will be inspected daily to collect emerging insects. Insects will be either preserved in ethanol or used to initiate colonies. Other aquatic plants growing in the same habitats as hydrilla will also be collected and held for insect emergence. Comparison of the species diversity of insects from hydrilla and those from other aquatic plants will provide a first indication of host specificity. If insects are located on various plants, then the specificity is not likely for hydrilla. The goal is to find an insect that is unique and specific to hydrilla for the least amount of non-target damage to other plants.

Preserved insects will be sent to world leading taxonomic experts for species level identification. Permits required to export dead insects from the African country where they were collected will be acquired. These permits will allow completion of requirements of US government regulations (U.S. Fish and Wildlife) to import the dead insects for scientific purposes.

Activity 4 Milestones

During the first year of the project at least 4 species of hydrilla herbivores will be collected and identified. A checklist of all insects reared from hydrilla will be prepared, and updated as the project progresses.

Activity 5: Develop rearing techniques and conduct initial screening.

Selected arthropods found feeding on hydrilla will be transported to laboratories at collaborating institutions to study their biology. Priority will be given to those agents found feeding exclusively on hydrilla. Rearing methods will be developed based on the biology of each candidate insect. Preliminary host range testing of colonized insect herbivores will be conducted in laboratories in Africa using hydrilla as well as other African Hydrocharitaceae as test plants. Permits to export natural enemies from Africa will be processed through the governments of Kenya, Burundi and Uganda depending on the location the insects were collected.

Activity 5 Milestones

A colony of at least one hydrilla natural enemy will be established at collaborating research laboratories in Africa within 3 years.

Activity 6: Importation of candidate agents.

Arthropods, which appear to be specific to hydrilla and cause measurable damage to plants, will be imported into quarantine in Florida (either the UF in Gainesville or the Fort Pierce Research Facility) for host range studies following approval of federal and state authorities.

Activity 6 Milestone and Measure of Success for the Project

State (FDACS) and federal (USDA/APHIS) import permits for at least 1 hydrilla herbivore will be processed within the 3-year project. At least one insect will be imported into a quarantine laboratory in Florida for host range and efficacy studies.

Schedule of activities for East Africa Effort


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