Quarterly Progress Report: Fourth Quarter FY 07

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EPA Demonstration Project on Hydrilla and Hygrophila in the Upper Kissimmee Chain of Lakes; EPA Grant #X796433105-1 (Hydrilla / Hygrophila Demonstration Projects - Osceola County, FL)
                                   
Project description and Goals:        

This project is a combination of studies that will result in improved water quality and natural habitat in Osceola County lakes.  Hydrilla and Hygrophila are exotic invasive weeds that are dominating the lakes aquatic ecosystem, shading out natural vegetation, reducing oxygen level, increasing the amount of sediments, and impacting flood control in lakes and canals. The project objectives are to evaluate the effectiveness of Experimental Use Permit herbicides, biological controls and application methods of herbicides in the treatment and control of Hydrilla and Hygrophila using small-scale fieldwork, large-scale field demonstrations and public outreach on the progress and findings.

Participants: 

Hilda Hatzell, EPA Project Officer, Region 4
Eleanor Foerste, Project Manager, Osceola County              

Reporting period:     

Fourth Quarter, EPA Fiscal Year 2007 July 1 – September 30, 2007

Project activities during reporting period: 

What work was accomplished for this reporting quarter?

Grant

• Took possession of the Grizzly Tracker Boat, Mercury Motor and Trailer that will be used in data collection for the grant.  Received Florida boat registration and numbers and permanent license plate for trailer.

Element 1

• Stewart Road Pond treated with Bispyribac-sodium and plant samples were collected and measured.
• Herbicide evaluations were conducted for phytotoxicity to hydrilla.
• Native plants were evaluated for sensitivity to flumioxazin.

Element 2

• Sample sites on Lakes Toho and Cypress were marked with GPS.  Water quality measurements were recorded and water samples were taken to be analyzed for ppb endothall.
• A large scale shallow water treatment was discussed by FDEP, SFWMD and FFWCC and will take place early in FY08 Q1.  The treatment effectiveness and impacts on tuber suppression will be monitored.

Element 3 Task 1

• Continued evaluation of new Mt formulations and production concepts.
• Discussions with large scale commercial fermentation companies to plan for scale-up of Mt dry material production.
• Outdoor mesocosm trials in NC initiated to support field trials.

Element 3 Task 2

• Samples of hygrophila were collected from several locations in the United States (Florida and Texas) for genetic analyses.
• Development of microsatellite primers for DNA analyses has been completed.
• Traveled to India to establish collaboration with overseas partners.
• Visited herbarium in Kolkata (Calcutta) to examine hygrophila specimens.
• Surveyed CABI’s pathogen collection for fungi associated with hygrophila

Element 3 Task 3

• Samples of hydrilla were collected from several locations in the United States, Africa and Asia for genetic analyses.
• Preliminary analysis of chloroplast DNA has been completed and a haplotype map constructed
• 19 lakes in Kenya, Burundi and Uganda have been sampled.  Hydrilla has been identified in 5 of the lakes.
• A large number of insects have been reared from hydrilla.  Many have been identified while others are still waiting identification.  By far the dominant group of insects are chironomid midges, with Dicrotendipes fusconotatus being the predominant species.  The trophic relationships of these midges to hydrilla are not yet certain.
• An MSc student has been recruited to work with the project in Uganda, and is funded from a USAID sponsored distance education program.

Element 4

• The website has been updated to provide visitors with more content and pictures of the grant activities.
• Developed abstracts for submission to local and national meetings.
• Brochures and webcards are in development to be used at events and meetings.
• Prepared newsletters and articles for local papers and trade publications.

Project spending for reporting period:

$73,091.52 was expended this quarter; therefore the remaining balance is $ 2,321,407.74.
(2,881,00.00 - 559,592.26) .

 

EPA Demonstration Project on Hydrilla and Hygrophila
in the Upper Kissimmee Chain of Lakes
EPA Grant   #X796433105-1

Reporting period: Fourth Quarter, EPA Fiscal Year 2007 July 1- September 30, 2007

Person Reporting: Sarah T. Berger, Osceola County Senior Biologist
Dr. William T. Haller, University of Florida, CAIP
Chris Mudge, Graduate Student, University of Florida

Element:  Element 1: Development and Testing of New Aquatic Herbicide

Project activities during reporting period:
The following activities were undertaken from July 1- September 30, 2007 in Osceola County, Florida:

1. Pond located at intersection of Pleasant Hill Road and Stewart Road was treated on June 27, 2007 with an EUP herbicide, Bispyribac-sodium.  Bispyribac-sodium was applied at 45 parts per billion (ppb) through long, weighted trailing hoses to assure uniform distribution.  Due to the fact that the pond is of uniform depth, one quarter of the total amount of herbicide needed for the pond was added to 80 gallons of water and four tanks were uniformly distributed in the pond.

2. Pond was revisited by Sarah Berger approximately every two weeks.  Plant heights were measured according to the standard operating procedure.  Briefly, the methods were:

• Paddled on kayak to middle of pond.
• Used pole device to grab hydrilla plants at root.
• 10-25 plants collected.
• Return to shore.
• Ten plants with roots were selected at random from group.
• Each plant was measured to 1/8 inch.
• Plant heights were recorded in field notebook.

3. Data collected was converted from inches to centimeters.

4. Mean and standard deviation were calculated using the following equations (Table 1, Figure 1):

= (Σx)/n      Sx = /(n-1)

 

5. Data:

WAT	MIN	MAX	MEAN	SD
0	179.705	235.585	209.4548	19.59492
2	72.0725	207.7875	146.5263	43.985
4	45.085	134.112	210.185	45.1997
6	39.6875	192.405	131.3498	46.2844
8	55.245	160.02	116.5225	43.23635
10	6.35	160.655	98.3298	43.77537

    Table 1: Plant height data

Figure 1. Plant Height vs. Time

6. Initial herbicide evaluations for phytotoxicity to hydrilla were conducted the past three months on dichlobenil, acifluorfen, bensulfuron-methyl, flumiclorac and amino pyralid at the Center for Aquatic and Invasive Plants (CAIP) in Gainesville, Florida.  One of these compounds was sufficiently active to pass to phase II, replicated testing.  Herbicide selectivity studies were conducted with emergent plants evaluating the EUP flumioxazin.  The herbicide was applied to foliage and as a submersed treatment.

7. At the CAIP, The emergent aquatic plants eleocharis, maidencane, pickerelweed, and sagittaria were evaluated for sensitivity to submersed and foliar flumioxazin treatments.  Two sprigs of each species were planted in a mixture of 2:1 potting soil:masonry sand in 3 L HDPE pots (17.1 cm diameter by 13.3 cm deep) amended with Osmocote (15-9-12) fertilizer at a rate of 1g kg-1 soil and placed in 95 L HDPE tubs filled with tap water (pH 7.5 at planting).  One pot of each species was grown for 4 wk in 95 L HDPE tubs placed in a shade house (30% shade).  Water level in the tubs was maintained at 25 cm, regardless of submersed or foliar herbicide treatment and pH remained at or near 7.5.  This experiment was a randomized design with five replicates (tubs).  Plants were grown for 1 mo, then flumioxazin was applied at 0, 50, 100, 200, 400, 800, and 1600 µg L-1 as a submersed treatment or at 0, 36, 72, 143, 286, 572, and 1144 g ha-1 a.i. plus a non-ion surfactant (0.25% v/v) as a foliar flumioxazin treatment.  Plants were treated using a forced air CO2-powered sprayer at an equivalent of 935 L ha-1 diluent and delivered through a single TeeJet® 80-0067 nozzle at 10 psi.  Plants were harvested 40 DAT and plant height from the soil surface to the tip of the tallest leaf was recorded, placed in a drying oven at 90 C for ca. 1 wk then weighed.  Results from the treatment are shown in the table below (Table 2)

 

Table 2. Effect of submersed flumioxazin treatment at pH 7.5 on emergent species dry weight and height 40 DAT in 95 L tubs in a shade house.
Dry Weight	EC50a (95% CIb)		Regression equation		r2
Eleocharis	559 (389-1009)		y = 13.7460e-0.00124x		0.92
Maidencane	259 (168-564)		y = 9.2236e-0.00268x		0.84
Pickerelweed	894 (598-1777)		y = 9.4660e-0.000775x		0.91
Sagittaria	15 (11-26)		y = 8.5266e-0.0448x		0.93
a - Effective concentration 50: EC50 = concentration of flumioxazin (µg L-1 a.i.) in water to reduce plant dry weight or height by 50%. Each value is a mean of two experiments with a total of 10 replications (pots). b - 95% CI = 95% Confidence Interval.

What Milestones have been met for your Element?

Element 1 Milestones and Measure of Success:

Element 1, Task 1 Milestone:

Hiring/training of personnel will occur in Year 1 and 3 to 4 treatment sites will be located for herbicide treatment the first and second quarter of year 2007.  An additional 3 to 4 sites containing different native aquatic plants will be treated annually (spring) of each year and plant communities monitored for up to 1 year following herbicide application.  Hiring completed April 16, 2007.  Training will be a continuing process.  Due to delayed approval of the QAPP, one site was treated and is still being evaluated this year.

Element 1, Task 2 Milestone:

In the first two years, 5-10 compounds will be tested annually for activity against hydrilla and hygrophila.  Concentration/exposure times (CET) will be conducted on these products with the greatest potential for future registration, as well as some of the older products (1-2 products per year).  Greenhouse and shadehouse studies (step 3 above), and phytotoxicity studies will be conducted on the greatest potential for future registration, which may be two to three products annually. Completed throughout year but testing still ongoing.

 

EPA Demonstration Project on Hydrilla and Hygrophila
in the Upper Kissimmee Chain of Lakes
EPA Grant   #X796433105-1

Reporting period: Fourth Quarter, EPA Fiscal Year 2007 July 1 – September 30, 2007

Person Reporting:  Dr. Michael D. Netherland

Element:  Element 2 Tasks 1 and 2: Evaluation of Currently Registered Herbicides for Control of Hydrilla
Element 2 Task 3: Evaluation of Currently Registered Herbicides for Control of Hygrophila

Project activities during reporting period:
Element 2 Task 1: Operational Treatments for Control of Hydrilla at the Very End of the 3rd Quarter:

The Florida Department of Environmental Protection (FDEP) and the South Florida Water Management District (SFWMD) conducted two large-scale endothall treatments in June 2007 to control hydrilla on Lakes Tohopekaliga (Toho) and Cypress, FL.  On June 19, 2007, liquid Aquathol was applied to Lake Toho via two helicopters on a calm sunny day.  A total of 8000 gallons was applied over 555 acres to achieve a target concentration of 3 mg/L in the treatment plots.  Hydrilla acreage in this area of the lake was estimated to cover over 4000 acres at the time of treatment.  Treatments blocks ranged in size from 40 to 80 acres. One 80-acre plot in Lake Toho was treated with a combination of diquat at 0.37 mg/L and endothall at 1.0 mg/L.  A total of 22 sample sites both within and outside the treatment blocks were established for Lake Toho.   Lake Cypress is approximately 4,500 acres in size and the hydrilla coverage in the northern 1/3rd of the lake was estimated between 800 and 1000 acres.  On June 21, 2007, a total of 2800 gallons of liquid Aquathol was applied via helicopter over 300 acres to achieve a target concentration of 3 mg/L within the treatment block.   A total 11 sample sites were located within and outside the treatment zones in Lake Cypress.  Water sampling for endothall residues, water quality, and efficacy are described below.

Ongoing Sampling Efforts in the 4th Quarter

Sample sites on Lakes Toho and Cypress were marked via the use of 2 separate Garmin Map 76 GPS units (Figures 2 and 3).  The waypoints were recorded as UTM coordinates.  The water depth, hydrilla coverage, and percent volume infested (height of hydrilla canopy in relation to the water depth) were recorded at each sample site on a weekly basis through 5 WAT.  Sample sites were visited on July 3, 5, 7, 9, 11, 17, 19, 24, and 31, and August 3rd, and September 10th during this Quarter.  Water quality measurements (temperature, pH, DO, and conductivity) were recorded at 3 depths in the water column with a calibrated YSI 556 meter at selected sample sites.  Water samples for endothall residues were collected during the first two weeks of July and efficacy assessments are ongoing.  All endothall residues for the June and July sample collections on lake’s Toho and Cypress have been analyzed.

Figure 2. Sample points on Lake Toho associated with June 2007 aerial endothall applications. Endothall residues, water quality and efficacy are assessed at each site.

Figure 3. Sample points on Lake Cypress associated with June 2007 aerial endothall applications. Endothall residues, water quality and efficacy are assessed at each site.

Efficacy monitoring for the large-scale endothall treatment conducted on Goblit’s Cove of Lake Toho and Lake Cypress in March 2007 is still ongoing.  All sample sites were visited once in July, August, and September, and monthly visits are planned to the end of this calendar year.  Hydrilla control in Goblit’s cove and the southern half of Lake Cypress remained good through 6 months after the initial application.  Water samples collected in conjunction with these treatments have been analyzed via an enzyme-linked immunoassay. 

Upcoming Treatments – Element 2 Task 2

A large-scale shallow water treatment (water depth is less than 4 feet in average depth) in the north end of Lake Toho was discussed during the 4th quarter of FY07, and it will be implemented early in the first quarter of FY08.  Treatment efficacy, water quality, impacts on tuber formation and tuber population dynamics, herbicide residue collection, and treatment longevity on this 450 acre treatment will be assessed for a 6 to 12 month period.

Hygrophila Mesocosm Evaluations: Element 2 Task 3

Hygrophila polysperma (hygrophila), collected from Boggy Creek in Osceola County was transported to the Center for Aquatic and Invasive Plants in Gainesville.  Sprigs of hygrophila were planted in 4 Liter pots and the plants rapidly established.  In late July, plants were placed in 95 L treatment tanks and exposed to the following treatments:  Diquat – 0.37 ppm, Diquat 0.18 ppm + Hydrothol 0.5 ppm, Diquat 0.37 + Aquathol 2.0 ppm,  Hydrothol 1.0 ppm, Triclopyr 2.0 ppm, and Flumioxazin 0.4 ppm.  Plants were exposed to the herbicide concentrations for 24 and 48 hours.  Following the herbicide exposures plants were moved to 900 L grow-out tanks.  Treatment efficacy was assessed at 6 weeks after treatment (WAT) and a final assessment will be conducted at 12 WAT (1st Quarter of FY08).   

What Milestones have been met for your Element?

Element 2, Task 1 Milestones and Measure of Success:

Fall of 2006 and Spring of 2007: An injector system will be constructed in the Fall of 2006 and treatments will be scheduled for November 2006.  A preliminary dye study will be conducted to provide a prediction of herbicide dispersion patterns by December 2006.  This short-term study will allow us to determine appropriate use rates and duration of the drip application.  If the initial demonstration is successful, other areas that can benefit from drip application technology will be evaluated, and further treatments will be scheduled in October through March in both 2007 and 2008. 
Residue, efficacy, and selectivity data collected during this effort will be written up as a manuscript and submitted for peer-review publication.  The project evaluates and documents, a little used technique for controlling hydrilla in flowing waters.  New information should lead to better long-term control at a reduced cost and with minimum applicator exposure.  If we demonstrate this technology to be superior to current strategies, we will continue to refine use rates and timing to provide optimal hydrilla control.

Response -

We did not receive approval from EPA to conduct work on this Element until March of 2007.  All sampling for the March treatments was conducted under a conditional approval of the QAPP.  Due to the low water flow conditions throughout the Kissimmee Chain and in Osceola County, we were not able to conduct any dye injection studies or operational treatments using an injector system.  In lieu of injectors, we evaluated large-scale demonstration treatments employing aerial liquid applications of the contact herbicide Aquathol.  We helped to plan the treatments during the 2nd and 3rd quarters of FY07, and our group conducted extensive follow-up sampling of 4 major herbicide applications on the Kissimmee Chain of Lakes.  The switch to liquid applications from traditional granular applications results in significant cost savings and reduced applicator exposure.  We continue to assess efficacy in all of these treatment areas to determine longevity of control.  Preliminary results of this work have been reported at the Aquatic Plant Management Society meeting in Nashville, TN, and the Florida Aquatic Plant Management Society meeting in St. Petersburg, FL.  Following collection of 1 year of post-treatment data for the various treatment sites, we will work on a manuscript for publication.      

Element 2, Task 2 Milestones& Measure of Success: 
In the fall of 2006, coordination between Osceola County managers and UF staff in the determination of areas in the County that would benefit from shallow water applications of endothall for control of hydrilla.  Evaluation of 2 to 5 shallow water operational applications will occur in the fall of 2006, the winter/spring of 2007, the summer of 2007 and the fall of 2007.  We plan to refine the application techniques and use rates and focus on demonstrating optimal treatment strategies will be addressed in 2008 and 2009.  The measure of success in this task is evaluating and providing information to Osceola County that will increase the efficacy and longevity of shallow water endothall treatments.

Response:  We did not receive QAPP approval to work on this task until May 2007.  At that point it was difficult to focus on shallow water applications, as hydrilla had spread to cover very large areas of the deeper open water of Lake’s Toho and Cypress.  From an operation’s perspective, these areas were deemed higher priority than the shallow zones.  It should be noted that the June treatment on the north end of Cypress could technically be classified as a shallow water application, as the average depth of the treatment zone was between 4 and 5 feet of water.  We continue to monitor hydrilla recovery in this area.  During the 4th quarter of 2007, treatment of shallow zones of Lake Toho were specifically discussed, and the current plan is for a large-scale application to be implemented in late October 2007 on 450 acres in the shallow area (average depth < 4 ft) of the north end of the lake.  We plan to monitor both short and long-term efficacy, impact of the treatments on tuber formation, and the pattern of residue dissipation and degradation both within and just outside the treatment zone.  Treatments of 3 ppm Aquathol alone will be compared against a combination of Aquathol at 1 ppm and Diquat at 0.37 ppm.

Element 2, Task 3 Milestones: 

Existing registered products and EUP products will be screened during years 1 and 2 of the project.  Products that show little potential for performance will be removed from further consideration.  Starting in year 2 and continuing through year 3, we will select the products that have the optimal efficacy and environmental characteristics for more intensive testing at the mesocosm scale.  Based on results from the mesocosm trials, field-testing will be initiated in year 2 or year 3.  Following data collection, reports will be provided on the optimal product or product combination that provides control of hygrophila in a high-flow environment.      
 
Response:

We did not receive QAPP approval to work on this task until May 2007.  In June, 2007 hygrophila was collected from Boggy creek in Osceola County and the plants were grown in culture at the Center for Aquatic and Invasive Plants in Gainesville, FL.  The hygrophila grew very well in culture and initial mesocosm studies were conducted in July 2007.  We screened six compounds under short-term exposure scenarios to determine if any of the herbicides have potential to provide control in situations where rapid dilution is expected.  Initial efficacy was evaluated at 6 WAT and the final evaluation will be conducted at 12 WAT (early in the first quarter of FY08).  Additional mesocosm trials are planned for FY08, and rates and exposures will be modified based on the findings of the initial trial.  Potential field sites in Osceola County have been identified, and we have discussed potential demonstrations of endothall and diquat combinations.   

 

EPA Demonstration Project on Hydrilla and Hygrophila
in the Upper Kissimmee Chain of Lakes
EPA Grant   #X796433105-1

Reporting period:     Fourth Quarter, EPA Fiscal Year 2007 July 1 – September 30, 2007

Person Reporting:   Mark A. Heilman   

Element:

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 activities during reporting period:

1. Lab Evaluations of Mt formulations and methods

Parallel studies with USDA and ERDC continue to evaluate new formulation and production concepts outside the formal scope of this grant program.  Four specific screening trials were conducted in laboratory aquaria mesocosms.

With respect to grant-supported trials, ERDC cooperator Dr. Judy Shearer completed a aquarium mesocosm study initiated in late June conducted in conjunction with late June field trial.  This study included interaction with fluridone aquatic herbicide (trade name Sonar) as an integrated component.  The results in Table 3 indicate better than 80% hydrilla biomass reduction at 20 mg/L Mt rate and improved injury in combination with 10 ppb fluridone.

  Table 3. Results from ERDC Study

2. Collaborative Research with NC State (originally Q1 and Q2)

Formal studies in this area with NC State University have only very recently begun due to various other commitments by NCSU (Dr. Rob Richardson) and time to coordinate with SePRO personnel.  As of September 2007, early studies of ecophysiology have begun at the SePRO RTC, and agreement with NCSU to get student effort on this project has been worked out for the fall.  As of this time, while delayed, SePRO has engaged NCSU as originally intended in project work plan. 

The early work in this area suggests that net photosynthesis, as monitored through dissolved oxygen changes in water around treated hydrilla tissue, may be a useful method of quickly detecting successful Mt infection for future evaluations.  Figure 4 shows hydrilla in shallow containers maintained in lab growth chamber.  The associated graph (Table 4) documents dissolved oxygen changes in a one-hour incubation of hydrilla tissue 48 hours associated with various Mt treatments.

Figure 4.  Hydrilla condition 48 hours after treatment with increasing rates of Mt

  

3. Pilot-scale commercial production

SePRO and its USDA collaborators have had detailed conversations over the last six months with multiple potential large-scale commercial fermentation companies to plan for scale-up of Mt dry material production to allow expanded field evaluations of the bioherbicide.  As described in original work plan, additional grant support from the FDEP Bureau of Invasive Plant Management will help support this effort.  Due to various administrative delays in contracting between USDA and FDEP, these funds have only been secured in the last 60 days.  Additionally, SePRO and USDA are continuing to evaluate best product formulate(s) to take into scale-up process.  As part of this process, SePRO has enlisted the assistance of a former Eli Lilly fermentation expert to consult in evaluating current process and providing suggestions on ways to improve production and scale-up protocols.  At this point, a scale-up decision is anticipated in the last calendar quarter of 2007 to support aspects of early 2008 testing.

                   Table 5. Results of outdoor tests of best Mt formulations

 

4. Outdoor testing of best formulations

On August 1, 2007, SePRO initiated an outdoor mesocosm trial at its NC facility to support field trial initiated August 2.  Table 5 (above) shows less injury versus late June ERDC study, likely due to static conditions of outdoor test tanks versus actively aerated ERDC aquaria.  At 5 ppb fluridone dose, results do not indicate interaction between Mt and fluridone in this trial.

Overall, indoor and outdoor mesocosm testing has documented Mt rate effects for comparison to small-scale field results.  Confirmation of bioherbicide interaction with fluridone has been achieved for one of the trials.  Differences between static and aerated test mesocosms suggests role of water exchange in transferring Mt infective propagules throughout treated hydrilla biomass.

5. Initiation of small field demonstrations in Osceola County

Enclosure studies in Osceola County ponds under the approved QAPP program were initiated May 25, June 21, and August 2.  The May 25th treatment was not successful due to either an enclosure method modification or heavy early June rain associated with Tropical Storm Barry.  Hydrilla biomass collection at one month after treatment showed no reductions from treatment (Table 6).

    Table 6. Hydrilla biomass one month after treatment

The June 21 (Table 7) demonstration effort examined a dry Mt formulation, unformulated material, and dry formulation in combination with fluridone herbicide (trade name Sonar).  Fluridone has been previously documented to interact synergistically with Mt bioherbicide.  This interaction had never been previously documented under field conditions.  Results of one-month and 2.5-month post harvests demonstrated this synergism as enclosures treated with Mt and Sonar showed improved reductions in hydrilla biomass versus enclosures treated with either product alone.  Unfortunately, results with Mt alone were not satisfactory and less than early spring treatment.  This was likely due to elevated water temperatures and other water quality stress associated with vertical stratification in mature hydrilla beds.  Mt bioherbicide does not grow well above 30 degrees Celsius, and concurrent water temperature data monitoring determined surface water temperatures exceeded this level for most of study period.  Visual observations also suggested that hydrilla injury was greater in shoot sections below hydrilla mat where ambient water temperatures were several degrees C cooler. 

   Table 7. Treatment results for June treatment.

On August 2, a follow-up trial was initiated to replicate the Sonar:Mt interaction with an effort to improve Mt performance by removing most of hydrilla canopy at time of study setup.   Shearers were used to remove the top 18 inches of hydrilla mat in enclosures one week prior to treatment.  The goal here was to improve water exchange within treatment plots and allow Mt product to settle into cooler zone where it would likely be more virulent.  Biomass harvest from one-month assessment of this treatment showed no significant treatment effects, and water temperature data suggested that pre-treatment canopy removal did not enhance mixing with enclosures.  In addition, dissolved oxygen monitoring suggests that since early August, subcanopy oxygen concentrations in the trial pond have dropped significantly to much less than 1 mg per liter.  Like almost all fungi, Mt is an aerobic organism requiring oxic conditions for growth.  The near-anoxic water conditions observed in the test pond suggest that insufficient oxygen may be present for virulent Mt growth.  With these observed field conditions, further pond demonstrations have been delayed until later in the fall pending cooling of pond surface waters.

Also, in mid September and early October, shoreline areas of one pond were also treated with a low dose of chelated copper herbicide to remove matted, algae-covered hydrilla and stimulate fresh hydrilla growth for later Mt bioherbicide treatment at this site.

What problems, if any, were encountered?

Most significant issue is with apparently unfavorable, summer-time environmental conditions in Osceola County pond sites for effective hydrilla control with Mt pathogen.  Water column stratification and related water temperature extremes is known to impact bioherbicide performance, and field evaluations have confirmed this for new formulated materials.

If a problem was encountered, what action was taken to correct it

August field study attempted mechanical canopy removal 1-week prior to Mt treatment to try to improve water column mixing.  This did not appear successful.  Also, overall pond dissolved oxygen decreased late in period to near anoxic levels below hydrilla canopy.  This condition also is unfavorable for bioherbicide use due to aerobic requirements of Mt pathogen, so further work has been delayed until fall and cooler water temperatures with higher dissolved oxygen.

• Is the project work on schedule?

The collaborative team is pushing hard to confirm an appropriate bioherbicidal formulation for scale-up production and expanded field evaluation in 2008.  Secondary data is also being generated for detailed temperature responses of organism for submission to EPA as part of a fall EUP request for 2008.  Overall, the scale of field testing is lagging slightly for several reasons.  First, the collaborative team has not determined best production protocols for cost-effective scale-up of production necessary for larger testing.  Additional consulting is hoped to resolve these questions in the next six months.  In parallel, further regulatory approval from EPA Biopesticide division will be needed for Experimental Use Permit allowing expanded testing.  SePRO will pursue this approval in next quarter in parallel with continued production evaluations.  In conclusion, overall scope of testing under grant remains on schedule but significant action will be needed in next two quarters to meet 2008 objectives for successful field evaluation of Mt bioherbicide.  

• Does the new schedule require a time extension

Based on field results, expanded field effort will be scheduled for fall and spring when environmental conditions appear more favorable for bioherbicide use.  This does differ some with original work plan that had more summer activities.  Summer field use in 2008 will be carefully evaluated based on site water temperature and general water quality favorable for good bioherbicide performance.  Scale of 2008 field evaluations and related schedule will be determined by scale-up and regulatory activity in next two quarters.

 

EPA Demonstration Project on Hydrilla and Hygrophila
in the Upper Kissimmee Chain of Lakes
EPA Grant   #X796433105-1

Reporting period:      Fourth Quarter, EPA Fiscal Year 2007 July 1 – September 30, 2007

Person Reporting:     James P. Cuda

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

Project activities during reporting period:

• Samples of hygrophila were collected from several locations in the United States (Florida and Texas) for genetic analyses.
• Development of microsatellite primers for DNA analyses has been completed.
• Traveled to India to establish collaboration with overseas partners.
• Visited herbarium in Kolkata (Calcutta) to examine hygrophila specimens.
• Surveyed CABI’s pathogen collection for fungi associated with hygrophila

What Milestones have been met for your Element?

Element 3, Task 2:  Milestones:

Activity 1 Milestone: 

Identify collaborating institutions during Year 1 – FY 2007.

A formal agreement with CAB International (CABI) was signed in late 2006.  In September 2007, traveled to Delhi, India, to attend planning meetings with CABI. 

A visit was also made to the Project Directorate of Biocontrol Control (PDBC) (an institute of the India Centre for Agricultural Research [ICAR]), in Bangalore in order to build a relationship of mutual benefit between University of Florida (UF) and PDBC.  UF does not have a MoU with PDBC in place , but the director, Dr. Rabindra, suggested that this project could be incorporated into the ICAR – CABI Workplan (CABI has an MoU with ICAR). This would help facilitate the potential future need to export to Florida any suitable biocontrol agents of hygrophila that are discovered. 

Activity 2 Milestone: 

Catalogue herbaria and literature records of target plants

A trip to India’s Central National Herbarium located in Kolkata, West Bengal, India, was made in September 2007.  In total, 64 specimens of hygrophila were examined and the locality information/ecological notes recorded.  The herbarium’s records indicated that hygrophila was collected from 12 Indian states, the majority of samples (26 of 64, or 41%) from the state of West Bengal in northeast India.  The earliest record was 1910 and at least one sample was collected at an altitude of 1200m.  Contact has been made with the Director of the Herbarium at Kew, London, to arrange a visit to examine the Hygrophila collection.  This will help determine distribution of H. polysperma in its centre of origin, since records date back to at least the 1800s.

Activity 3 Milestone: 

Identify extant populations of target plants.

The first population of Hygrophila sp. was discovered on the Yamuna River that flows around Delhi in September 2007 and georeferenced for possible sampling of potential natural enemies at a later date.  

Activity 4 Milestone: 

Collect hygrophila for molecular studies. 

Collection of hygrophila samples from Florida, Texas and India for DNA analyses is in progress.  Development of microsatellite primers for DNA analyses was completed during the last quarter.  Techniques are being developed to extract and sequence chloroplast DNA.

Activity 5 Milestone: 

Rear natural enemies from target plants, and develop rearing methods for selected natural enemies.

Visited the CABI Fungal Collection to look at records of fungi from Hygrophila species during this quarter.  The following fungi were recorded from unidentified species of Hygrophila:  Aecidium manilense (= Puccinia cacao) (Philippines), Uredo hygrophilae   (Philippines), Uredo hygrophiliicola (India, Burma), and Blastospora hygrophile (India).

Activity 6 Milestone: 

Import selected natural enemies into Florida quarantine laboratories.

Permits to import hygrophila and hydrilla samples from India have been approved by USDA, APHIS during this quarter.  However, we are not close to importing any insects at this time.

Is the project work on schedule?

Though efforts are being made, it is likely that establishing sub-contracts with Indian collaborators (PDBC) could result in some delays in accomplishing project goals and objectives due to complex sub-contracting regulations at the University of Florida

 

EPA Demonstration Project on Hydrilla and Hygrophila
in the Upper Kissimmee Chain of Lakes
EPA Grant   #X796433105-1

Reporting period:      Fourth Quarter, EPA Fiscal Year 2007 July 1 – September 30, 2007

Person Reporting:     William A. Overholt

Element:  Element 3 Task 3 Collaborative effort to search for natural enemies of hydrilla in East Africa (Project Ongoing)

Project activities during reporting period:

• What work was accomplished for this reporting quarter?

• Samples of hydrilla were collected from several locations in the United States, Africa and Asia for genetic analyses.
• Preliminary analysis of chloroplast DNA has been completed and a haplotype map constructed
• 19 lakes in Kenya, Burundi and Uganda have been sampled.  Hydrilla has been identified in 5 of the lakes.
• A large number of insects have been reared from hydrilla.  Many have been identified while others are still waiting identification.  By far the dominant group of insects are chironomid midges, with Dicrotendipes fusconotatus being the predominant species.  The trophic relationships of these midges to hydrilla are not yet certain.
• An MSc student has been recruited to work with the project in Uganda, and is funded from a USAID sponsored distance education program.

What Milestones have been met for your Element?

Element 3, Task 3:  Milestones:

Activity 1 Milestone: 

All agreements will be signed between collaborating institutions during Year 1 – FY 2007.
This milestone was achieved in late 2006 with agreements being signed with the University of Miami for molecular genetic work, and with the International Center of Insect Physiology and Ecology (ICIPE) in Nairobi, Kenya for exploration for hydrilla herbivores in East/Central Africa.  In turn, ICIPE signed agreements with the National Institute for the Environment and Conservation of Nature in Burundi and the National Fisheries Resources Research Institute in Uganda.  In August 2007, the main genetics collaborator, Dr. Dean Williams accepted, a position at Texas Christian University.  Thus, the agreement with the University of Miami is being transferred to Texas Christian University (to be completed in October, 2007).

Activity 2 Milestone: 

Database and maps produced and posted on the worldwide web (Dr. Overholt, please send this information to Element 4 coordinator).  The first maps will be available within the first month of the project. 

Maps of hydrilla collections have been posted on the website of the UF/IFAS Biological Control Research and Containment Laboratory (http://bcrcl.ifas.ufl.edu/Invasive_Weeds/Hydrilla/Hydrilla%20Reports.htm).  A database of hydrilla collections is also maintained at the same website (http://bcrcl.ifas.ufl.edu/6_Hydrilla_Surveys_HomePage.htm).

Activity 3 Milestones:

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

A total of 329 hydrilla samples have been collected and provided to Dr. Dean Williams at Texas Christian University of genetic analyses, including 26 from Uganda, 12 from Burundi, 231 from Florida, 10 from Louisiana, 40 from Texas, 5 from Maryland and 5 from Virginia.  Additionally, Mr. Paul Madeira from the USDA/ARS has provided 74 samples from a number of locations including China, India, Taiwan, New Zealand, Korea, Panama, Nepal, Indonesia, Pakistan, Japan, Poland, Burundi and several locations in the United States.

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

Techniques have been developed to extract and sequence chloroplast DNA, and a preliminary haplotype map has been constructed (posted here). Extraction of DNA for microsatellite analyses is underway.

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

This task is underway with cpDNA, but not yet started with microsatellite DNA

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.  

A number of insects have been reared from hydrilla, including several species of midges.  Some have been identified (see June 2007 report at http://bcrcl.ifas.ufl.edu/Invasive_Weeds/Hydrilla/Hydrilla%20Reports.htm), while others are still waiting identification.  However, the trophic relationships of these insects to hydrilla have not been confirmed.  The most abundant species we’re found, Dicrotendipes fusconotatus, is thought to be a ‘scraper’ that feeds on plankton/algae while sitting on the surface of hydrilla stems/leaves.  Two of the species collected (Polypedilum wittei and Polypedilum dewulfi) have been previously reported as herbivores of hydrilla, but while we have reared Polypedilum species from most of our collections, they have not, to date, been encountered in the large numbers reported during previous surveys."

Activity 5 Milestones: 

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

We have not yet accomplished this task.  Efforts are underway to colonize Polypedilum wittei and/or Polypedilum dewulfi in Uganda and Burundi.

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.

Permits to import any Diptera or Coleoptera associated with hydrilla from Africa have been approved during this quarter.  However, we are not close to importing any insects at this time.

 

EPA Demonstration Project on Hydrilla and Hygrophila
in the Upper Kissimmee Chain of Lakes
EPA Grant   #X796433105-1

Reporting period:      Fourth Quarter, EPA Fiscal Year 2007 July 1 – September 30, 2007

Person Reporting:     Dr. Tina M. Bond, Osceola County Extension
                                    Amy Richard, Center for Aquatic and Invasive Plants

Element:  Element 4 Demonstration and Outreach

Project activities during reporting period:

• What work was accomplished for this reporting quarter?
  • Attended a variety of meetings including
    • Hydrilla Interagency
    • Kissimmee Basin Modeling Operating Study
    • Osceola County Lakes Management Advisory Committee
    • Met with various agrichemical representatives to discuss their products and potential effects on invasive aquatic weeds
    • Met via the internet with the South Florida Water Management District titled ‘Water Summit 2007’
    • Overall Advisory Committee Meetings for the Extension Office
    • Natural Resources Committee Meetings for the Extension Office
  • Provided updates for the Hydrilla and Hygrophila Demonstration Project website: http://plants.ifas.ufl.edu/osceola/index.html
  • Updates include:
    • Bios of project leaders
    • QAPP and Quarterly Reports uploaded
    • Recent trip to Africa
    • Herbicide information and links to product labels and MSDS
    • Events and other related information
    • A link from the CAIP website to the Hydrilla Grant website was recently added.
    • Provided all written text and the majority of pictures and other content to the Hydrilla and Hygrophila Demonstration Project website
  • Participated in an educational effort at Deseret Ranch, assisting the 4-H Extension Agent
    • Worked with youth on understanding invasive plants
    • Played a hydrilla game with youth in order to show them how hydrilla can effect native plants and how biocontrols can help manage invasive plants

 
                                  Tina Bond and the Kids at Deseret Ranch   

        The Invasive Plant Game

• Submitted an article to Aquatics Magazine, the professional publication for the Florida Aquatic Plant Management Society. Scheduled for the Fall 2007 Issue.
• Continue to survey and tour the Kissimmee Chain of Lakes to keep up to date on the status of hydrilla and hygrophila growth and spread, as well as note problem areas and areas of residential concern.

  
Lake Cypress, hydrilla

Brick Lake, SFWMD treatment

• Assisted with an information factsheet that was submitted to the Osceola County Board of County Commissioners dealing with the status of aquatic weed control in the Kissimmee Chain of Lakes.  This occurred due to residential inquiry at a BOCC meeting in September.
• Registered for Great Outdoor Days; http://greatoutdoordays.com/
• Visited with SePRO to review treatments of Mt fungus in ponds located on Route 192 in Kissimmee, FL.
• Submitted an abstract to the Weed Science Society of America to present information on the grant.
• Interviewed for the Aquatics position with the University of Florida in order to be recognized as an Extension Agent within the University system.
• Attended New Agent Orientation at the University of Florida in Gainesville, FL.
• Attended the Extension Professional Association of Florida annual meeting in St. Augustine, FL.
• Submitted an article to the Celebration Newsletter.  Published September, 2007.
• Designed a brochure for events, tradeshows, etc. that will inform people about the grant and aquatic invasive plants.  Ordered end of September, expected arrival Mid-October.
• Continue to collect information for the website and programs to be taught.
• Prepared presentation on the grant for the Lakes Advisory Committee Meeting.
• Invited speaker at the August meeting of Osceola County’s Lakes Management Advisory Committee.
• Developed plan to incorporate the Lakes Management Advisory Committee as the Aquatics Grant Committee in order to obtain feedback on educational programs needed in the community.
• Inspected potential treatment locations for the grant.  The areas were suspected to have hydrilla and/or hygrophila, depending on the location of the water body.
• Collected information for Kimberly Lawrence, former Lakes Manager for Osceola County, on hydrilla growth and reproduction.  This information was used for a meeting that involved residents and county commissioners regarding Buenaventura Lakes area retention ponds.
• The grant was inconspicuously mentioned in an Orlando Sentinel article titled “Lakes Languish in Invasive Weeds”.  I was asked to provide information on the grant for the article, as was the FDEP, SFWMD, and the FWC for management and operations information.
• Developed presentations for the Florida Aquatic Plant Management Society and North American Lakes Management Society Annual Meetings where I will be presenting information on the hydrilla and hygrophila grant.

What Milestones have been met for your Element?

Element 4 Milestones and Measure of Success:

Element 4 Task 1 Milestone:

Year 1:  Kick-Off Event – introduced the public and local government officials to the goals of the project. Completed April 21, 2007.

CAIP staff registered for and are preparing for another public awareness event scheduled for October 27-28th in Kissimmee (GREAT OUTDOORS DAYS). Public awareness banners and educational materials will be displayed and distributed at the event.  Amy Richard and Rob Horsburgh will be staffing the booth at the event.

Element 4, Task 2 Milestone:

A draft outline, flowchart and master style sheets for the website will be completed by the end of December 2006. The website itself will be launched before the end of March 2007.  Completed April 20 to coincide with the Kick-Off Event.  Usability studies were not conducted in 2006 because the aquatics agent position was vacant until January, 2007.

Additional written material and images that were being edited and formatted (html) during the previous quarter are now posted/published on the Osceola Demonstration Project website. The additions total 32 webpages. These new documents include:

• Additional links page
• A biography page for project leaders/participants, miscellaneous images of Lake Toho, Channeled apple snail eggs, boating & recreation, etc.
• “East African Diaries" by R. Copeland, detailing his research trip to find new biological control agents for Hydrilla.
• Herbicide label information and new herbicide information
• Hydrilla Management Plan
• Listing of Prohibited Aquatic Plants
• Permit Conditions
• Quarterly reports from 2006 and 2007
• Photo/text page documenting Hydrilla Kick-off Event (4/21/07)
• Photo/text page documenting St. Cloud Extravaganza  (3/10/07)
• Photographs were taken in the Osceola County area (Lake Toho) and are currently being edited for integration into the site and more visual appeal to the non-scientific public.

The report “Project Report: Lake Toho & Lake Cypress Herbicide Treatments” is currently being edited and prepared for web publication.

Element 4, Task 3 Milestone:

A search of the current APIRS database will be done for any materials that would be pertinent to this project and tagged for easy location by researchers and the general public; to be accomplished by the end of December 2006.  Completed by the Center for Aquatics and Invasive Plants. This is available on the website.  Materials provided by project researchers will be added to the database within three weeks of receipt.  Completed August, 2007.  We did not have information from the researchers to add to the website until this time because the QAPP for the project had not been approved until May 2007.  Content for the website was mainly from Element 4 and Element 3, Task 3 activities, QAPP, Quarterly Reports, herbicide information, and other information.

Annotated citations to the scientific literature pertaining to hygrophila and hydrilla management in Osceola County and Kissimmee water bodies are listed online in the Osceola County Hydrilla & Hygrophila Demonstration Project website under Research Publications from the APIRS Database. Currently there are 17 citations listed. New citations will be posted as they are received. Other citations from the APIRS collection that could provide additional information on plant management in Florida will be posted as they are located and evaluated.).

Element 4, Task 4 Milestone:

Three introductory displays will begin at beginning of October 2006 and completed by the end of that same quarter (December 2006). Large, stand up display banners are used a various meetings and events.  They depict certain scenes with invasive plants.  Another set of displays can be produced by the end of December 2007, for use in various spring and garden festivals in early 2008.  We are a little behind on the making of displays because the aquatics agent began working on the project in January, 2007.

The large format public information displays used in recent public events are currently under review for ideas on developing future materials. New displays are in the planning phase for completion by the end of December 2007. 

Element 4, Task 5 Milestones:

Eleanor Foerste and Amy Richard have taken the lead on curriculum.  We have provided the Osceola County School District, Science Resource Specialist with a list of materials needed that make up a kit that will help them sample water bodies on school properties.  The materials (kits) have been purchased, training was held to instruct teachers on how to use the kits, and teachers have kits to use in their classrooms.  This occurred in August, 2007.  Materials will be developed, field tested, and made available by the end of June 2008 and presented to school teachers for use in the fall 2008.

CAIP staff made contact with a small number of science teachers in Osceola County and have provided them with materials and lessons to test in their classroom this year. Amy Richard will be meeting with these teachers sometime in January 2008 to assess the effectiveness of the lessons and activities being used in the classroom.

Element 4, Task 6 Milestone:

Development of demonstration and outreach strategies to help stakeholders understand and minimize the spread of hydrilla and hygrophila.  This would include printing of educational materials and items to be distributed at meetings and Field Day demonstrations.  This could include quick tips and fact sheets on hydrilla and hygrophila that direct clients and applicators to the website for additional information.  Examples could include laminated boater/angler quick ID cards, key chain floats, boat trailer stickers to remind boaters to clean trailers of plant debris, etc.  The lakes manager, Sherry Burroughs, and Element 4 coordinator are working on a two-part workshop that will initially include residents of Osceola County.  The meeting will determine lake issues plaguing the Osceola County residents.  The question and answer session will provide information that can then be relayed to government agencies and university faculty who can address these important questions.  After the agencies and university faculty have viewed the information, a second workshop with residents and agencies will be held.  In Osceola County, many residents do not understand why hydrilla is such a problem.  Their perception is that the government agencies are doing nothing to manage aquatic plants in the lakes, but this is far from what is occurring in the field.  We hope that this informative session will educate residents and other interested parties about how aquatic plants are managed by the different agencies in the county.  The workshop will also provide information regarding the biology and other facts surrounding this complex issue. Develop and implement at least 2 additional strategies to help demonstrate and educate stakeholders regarding hydrilla and hygrophila management strategies.

• Full color informational brochures about hydrilla and hygrophila, and the Demonstration project have been designed and printed and will be distributed at the next public event (late October). They will also be available at the Osceola County Extension office and other venues.
• Additionally, a set of “web cards” for both hydrilla and hygrophila have been produced and printed for handing out at public venues. The cards feature images of the plants and include the Demonstration Project web address, so the public can refer to the website, should they need more information.

Element 4, Task 7 Milestones and Measures of Success:

Year 1, 2, 3 and 4:  Prepare at least one poster and one research paper each year, sharing research progress and promising new hydrilla and hygrophila management technology.  We have not been able to submit research papers due to the delayed QAPP approval, hence delayed start of research. A banner for the Hydrilla and Hygrophila project was developed and displayed at the Kick-Off Event in April.

 

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East African Diaries

India Trip Report