NCG560067_Application_20240412

S1ATE
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ROY COOPER •i ' �a
Governor d�
ELIZABETH S.BISER
Secretary
RICHARD E.ROGERS,JR. NORTH CAROLINA
Director Environmental Quality
April 15, 2024
BlueGreen Water Technologies, Inc.
Attn: Eyal Harel
301 S Hill Village Ste LL200 #452
Pittsburgh, PA 15241
Subject: Permit Application
Application No. NCG560067
BlueGreen Water Technologies
Hyde County
Dear Applicant:
The Water Quality Permitting Section acknowledges receipt of your application for a new NPDES WW permit, including
supporting documentation and your check number 1434 in the amount of $100.00 as payment of the application fee.
These items were received in our offices on April 12, 2024. Your application will be assigned to a permit writer within
the Section's NPDES WW permitting branch. The permit writer will contact you if additional information is required to
complete evaluation of your application. Your timely and direct response to any such request will help to expedite the
review process. Please note that acceptance of the application does not guarantee a NPDES permit will be issued for the
proposed activity. A permit will only be issued following a complete review of the application, concluding the proposed
discharge is allowable per applicable statutes and rules.
Information regarding the status of your renewal application can be found online using the Department of Environmental
Quality's Environmental Application Tracker at:
https://www.deq.nc.gov/permits-rules/environmental-application-tracker
If you have any additional questions about the permit, please contact the primary reviewer of the application using the
links available within the Application Tracker.
Sinc el qoi
Wren edford
Administrative Assistant
Water Quality Permitting Section
cc: Jessica Frost, Scientific Director
ec: WQPS Laserfiche File w/application
D_E Q_J/ North Carolina Department of Environmental Quality I Division of Water Resources
v Washington Regional Office 943 Washington Square Mall Washington North Carolina 27889
ouon /` 252 946 6481

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Division of Water Resources FOR AGENCY USE O W.Y
A„y r' A` National Pollutant Discharge Elimination System (NPDES) Date Received
Year Month Ikiy
Application for Coverage Under aoaq 04 �a
General Permit NCG560000 Cern'mite or C,werage
tnrtro^mentur ouur. . Pesticide Applications a a I
ppliti N C C. 5 t�
Check# Amount
CW
NOTICE OF INTENT �� � i0o�
A,,;gt,�To: t RECEIVED
Required by I�:\ NC.\C 02H.0127fd1;term definition see 15,E NCAC 02H-0103(19)
(Please prim or type)
I) Mailing address of applicant'(address to which all correspondence should be mailed): APR 12 2024
Company Name BlueGreen Water Technologies
Owner Name Eyal Harel, Moshe Harel c/o Jessica Frost
K% DEQ/DWH/NPDES
Street Address 301 South Hills Village Ste LL200#452
City Pittsburgh State PA ZiP Code 15241
Telephone# [phone] Fax# NA
Cell# 941-224-2218 Email jessica.frosthbluegreenwatertech.com
'Applicants generally include both ti)the entity with control over the financing for,or the decision to perform pestc;de app,catuons. including the
ability to modify those decisions,that results in a discharge to waters of the State and(2)the entity with day-to-day operational control of or who
performs activities(e.g.,the application of pesticides)that are necessary to ensure compliance with the permit(e.g.,they are authorized to direct
workers to carry out activities required by the permit or perform such activities themselves).
2) Description of Discharge: [Required by 15A NCAC 02H .0105(c)(I)]
a) For what type[s]of pesticide-related discharge are you requesting coverage?
❑ Mosquito/flying insect pest control Acres:
(adulticide applications only)
0 Aquatic Weed/Algae control Acres: 756
❑ Aquatic Weed/Algae control Linear miles:
❑ Aquatic Nuisance Animal Control Acres:
❑ Aquatic Nuisance Animal Control Linear miles:
❑ Forest Canopy Pest Control Acres:
❑ Intrusive Vegetation Control Linear miles:
3) Have you prepared a Pesticide Discharge Management Plan? CS Yes ❑ No
'certification and signature shall be completed on the following page'
Page 1 of 2 Revised Jan 2019
L -

NCG560000 New Application
CERTIFICATION
I certify that I am familiar with the information contained in this application and that to the
best of my knowledge and belief such information is true, complete,and accurate.
Printed Name of Person Signing: Jessica Frost
Title: Scientific Director
(Please review 15A NCAC 02H .0106(0 for authorized signing officials),/ ,, August 16, 2023
(Signature of Applicant) (Date Signed)
North Carolina General Statute 4 43-215.6B provides that:
Any person who knowingly makes any false statement representation, or certification in any
application, record, report, plan, or other document filed or required to be maintained under this Article
or a rule implementing this Article;or who knowingly makes a false statement of a material fact in a
rulemaking proceeding or contested case under this Article; or who falsifies, tampers with, or knowingly
renders inaccurate any recording or monitoring device or method required to be operated or maintained
under this Article or rules of the Commission implementing this Article, shall be guilty of a Class 2
misdemeanor which may include a fine not to exceed ten thousand dollars($10,000). 18 U.S.C.Section
1001 provides a punishment by a fine or imprisonment not more than 5 years,or both, for a similar
offense.
Application must be accompanied by a check or money order for$100.00[per G.S. & 143-215.3(a)(lb)]
made payable to:
NC DEQ
♦ ♦ ♦ ♦ ♦
Mail this application and the fee payment to:
NC DEQ/DWR/NPDES
1617 Mail Service Center
Raleigh,North Carolina 27699-1617
Attn: Charles Weaver
ELECTRONIC SUBMISSIONS:
If you wish to complete and submit this application electronically,submit it to: [e-mail]
Your application can be received and reviewed electronically. However,the Certificate of Coverage(CoC)cannot
be issued until the application fee is received per G.S. 143-215.3(a0(I b).
ELECTRONIC RECEIPT OF CoC
Do you wish to receive your CoC electronically? ®Yes-CoC will be sent to the e-mail address provided above.
❑No-CoC and a copy of permit NCG560000 will be sent via USPS.
Page 2 of 2 Revised Jan 2019

United States Office of Water EPA Form 3510-1
Environmental Protection Agency Washington,D.C. Revised March 2019
Water Permits Division
..EPA Application Form 1
General Information
NPDES Permitting Program
Note: All applicants to the National Pollutant Discharge Elimination System (NPDES)permits
program,with the exception of publicly owned treatment works and other treatment works treating
domestic sewage,must complete Form 1. Additionally,all applicants must complete one or more of the
following forms: 2B,2C, 2D, 2E,or 2F. To determine the specific forms you must complete,consult the
"General Instructions"for this form.

Paperwork Reduction Act Notice
The U.S. Environmental Protection Agency estimates the average burden to collect information
and complete Form 1 to be 2.9 hours for new applicants and 0.9 hours for applicants renewing
existing permits. This estimate includes time to review instructions, search existing data sources,
gather and maintain the needed data, and complete and review the collection of information.
New respondents must also prepare a topographic map. Send comments about the burden
estimate or any other aspect of this collection of information to the Chief, Information Policy
Branch(PM-223),U.S. Enviromnental Protection Agency, 1200 Pennsylvania Avenue,NW,
Washington,DC 20460, and to the Office of Information and Regulatory Affairs,Office of
Management and Budget, 725 17th Street, NW, Washington, DC 20503,marked"Attention:
Desk Officer for EPA."

DESCRIPTION OF NPDES PERMIT APPLICATION FORMS CONTENTS OF FORM 1 PACKAGE
The application forms for individual National Pollutant Discharge Form 1—General Instructions
Elimination System(NPDES)permits include the following: Form 1—Line-by-Line Instructions
Form 1—General Information(included in this package). Form 1—Activities That Do Not Require Permits
Form 2—Forms Based on Facility or Activity Type(not included Form 1—Glossary
in this package): Form 1—Application
2A. New and Existing Publicly Owned Treatment Works
2B. Concentrated Animal Feeding Operations and
Concentrated Aquatic Animal Production Facilities
2C. Existing Manufacturing,Commercial, Mining,and
Silvicultural Operations
2D. New Manufacturing,Commercial,Mining,and Silvicultural
Operations That Have Not Yet Commenced Discharge of
Process Wastewater
2E. Manufacturing,Commercial,Mining,and Silvicultural
Facilities Which Discharge Only Nonprocess Wastewater
2F. Stormwater Discharges Associated with Industrial Activity
2S. New and Existing Treatment Works Treating Domestic
Sewage
FORM 1—GENERAL INSTRUCTIONS
Who Must Apply for an NPDES Permit? Upon your request,and based on information supplied by you,
With the exceptions described in"Form 1—Activities That Do Not EPA or the authorized NPDES state will determine whether you
Require Permits,"the federal Clean Water Act(33 U.S.C. 1251 et are required to obtain a permit for a particular facility or activity.
seq.)prohibits any person from discharging pollutants into waters Be sure to contact EPA or your state if you have a question.
of the United States without first having been issued a permit Form 1 collects general information only.You must also complete
under the NPDES program. a more detailed application based on your proposed discharge
Who Must Complete Form 1? activity,as follows:
All applicants,other than publicly owned treatment works •
If your facility is a concentrated animal feeding operation
(POTWs)and treatment works treating domestic sewage or a concentrated aquatic animal production facility,you
(TWTDS),must submit Form 1.If you operate one of the must also complete Form 2B.
following facilities,you must submit Form 1:concentrated animal • If your facility is an existing manufacturing,commercial,
feeding operations and aquatic animal production facilities: mining,or silvicultural facility that currently discharges
manufacturing,commercial,mining,and silvicultural operations: process wastewater,you must also complete Form 2C.
or other industrial facilities. • If your facility is a new manufacturing.commercial,mining.
At the state level.either the U.S. Environmental Protection or silvicultural facility that has yet to commence discharge of
Agency(EPA)or an approved state agency administers the process wastewater.you must also complete Form 2D.
NPDES permit program.If you are located in a jurisdiction in • If your facility is a new or existing facility(including
which an EPA regional office administers the NPDES permit manufacturing,commercial,mining,and silvicultural
program,you should use Form 1 and all other applicable forms facilities)that discharges only nonprocess wastewater,
described in these instructions.If you are located in a jurisdiction you must also complete Form 2E.
where a state administers the NPDES permit program,contact • If your facility is a new or existing facility whose discharge
the state to determine the forms you should complete.States is composed entirely of stormwater assnciated with industrial
often develop their own application forms rather than use the activity—excluding discharges from construction activity
federal forms.See http:/twww.epa.gov/npdes/npdes-state- under 122.26(b)(14)(x)or(b)(15)—you must also complete
program-information for a list of states that have approved Form 2F.If the discharge is composed of stormwater and
NPDES permit programs and those that do not. non-stormwater,you must complete Form 2F and you must
Exhibit 1-1 (see end of this section)provides contact information also complete Forms 2C,2D,and/or 2E.as appropriate.See
for each of EPA's 10 regional offices.Since the exhibit's content is Form 2F's instructions for further details.
subject to change,consult EPA's website for the latest information:
http://www.epa.gov/aboutepa#regional.
1-1

FORM 1—GENERAL INSTRUCTIONS CONTINUED
Where to File Your Completed Form Completion of Forms
• If you are in a jurisdiction with an approved state NPDES Print or type in the specified areas only.If you do not have enough
permit program.file according to the instructions on the state space on the form to answer a question,you may continue on
forms. additional sheets,as necessary,using a format consistent with the
• If you are in a jurisdiction where EPA is the NPDES form.
permitting authority(i.e.,the state is not an NPDES- The NPDES permitting authority could consider your application
authorized state),mail the completed application forms to the incomplete if you do not provide an answer(or indicate"NA"for
EPA regional office that covers the state in which your facility "not applicable")for all questions on Form 1 and the applicable
is located(see Exhibit 1-1). Form 2.
When to File Your Completed Form Provide your EPA Identification Number from the Facility Registry
Because of statutory and regulatory requirements,the deadlines Service.NPDES permit number,and facility name at the top of
for filing applications vary according to your facility or activity type each page of Form 1 and any attachments.If your facility is new
and the type of permit you need.The various permit application (i.e.,not yet constructed),write or type"New Facility'in the space
deadlines are listed in Exhibit 1-2 at the end of this section. provided for the EPA Identification Number and NPDES number,
If you do not know your EPA Identification Number,contact your
Fees NPDES permitting authority.See Exhibit 1-1 for contact
EPA does not require applicants to pay a fee for applying for information.
NPDES permits. However,states that administer the NPDES Do not leave any response areas blank unless the form directs
permit program may charge fees.Consult with state officials for you to skip them.If the form directs you to respond to an item that
further information. does not apply to your facility or activity,enter"NA"for"not
Public Availability of Submitted Information applicable"to show that you considered the item and determined
a response was not necessary for your facility.
EPA will make information from NPDES permit application forms
The NPDES permitting authority will consider your application
available to the public for inspection and copying upon request.
You may not claim any information on Form 1 (or related complete when it and any supplementary material are received
attachments)as confidential. and completed according to the authority's satisfaction.The
NPDES permitting authority will judge the completeness of any
You may make a claim of confidentiality for any information that application independently of the status of any other permit
you submit to EPA that goes beyond the information required by application or permit for the same facility or activity.
Form 1.If you do not assert a claim of confidentiality at the time
you submit your information to the NPDES permitting authority,
EPA may make the information available to the public without
further notice to you. EPA will handle claims of confidentiality in
accordance with the Agency's business confidentiality regulations
at Part 2 of Title 4 of the Code of Federal Regulations(CFR).
1-2

FORM 1—GENERAL INSTRUCTIONS CONTINUED
Exhibit 1-1.Addresses of EPA Regional Contacts and Covered States
REGION 1 ! REGION 6
U.S.Environmental Protection Agency,Region 1 U.S.Environmental Protection Agency,Region 6
5 Post Office Square,Suite 100,Boston,MA 02109-3912 - 1445 Ross Avenue,Suite 1200,Dallas,TX 75202-2733
Phone:[phone];toll free:[phone] Phone:[phone];toll free:[phone]
Fax [phone] Fax:[phone]
Website:http 1/www epa gov/aboutepa/epa-region-l-new-eneland Website:http-//www epa.gov/aboutepalepa-region-south-central
Covered stales:Connecticut,Maine,Massachusetts,New Hampshire,Rhode Covered states:Arkansas,Louisiana,New Mexico,Oklahoma,and Texas
Island,and Vermont
REGION 2 REGION 7
U.S.Environmental Protection Agency,Region 2 U.S.Environmental Protection Agency,Region 7
290 Broadway,New York,NY 10007-1866 11201 Renner Boulevard,Lenexa,KS 66219
Phone:[phone];toll free:[phone] Phone:[phone];toll free:[phone]
Fax:[phone] Website:http.r/www.epa.qou/aboutepa/epa-region-7-midwest
Website:http//www epa.gov/aboutepa/epa-region-2 Covered states:Iowa,Kansas,Missouri,and Nebraska
Covered states:New Jersey,New York,Virgin Islands,and Puerto Rico
REGION 3 • REGION 8
U.S.Environmental Protection Agency,Region 3 U.S.Environmental Protection Agency,Region 8
1650 Arch Street,Philadelphia,PA 19103-2029 1595 Wynkoop Street,Denver,CO 80202-1129
Phone:[phone];toll free:[phone] - Phone:[phone];loll free:[phone]
Fax:[phone] Fax:[phone]
Website:http i w.v,v epa govlaboutepa/epa-region-3-mid-atlantic Website:http!/www epa goviaboutepa/epa-region$-mountains-and-plans
Covered states:Delaware,District of Columbia,Maryland,Pennsylvania,Virginia, Covered states:Colorado,Montana,North Dakota,South Dakota,Utah,and
and West Virginia Wyoming
REGION 4 REGION 9
U.S.Environmental Protection Agency,Region 4 U.S.Environmental Protection Agency.Region 9
San Nunn Atlanta Federal Center • 75 Hawthorne Street,San Francisco,CA 94105
61 Forsyth Street,SW,Atlanta,GA 30303-8960 Phone:[phone];toll free:(866)EPA-WEST
Phone:[phone];toll free:[phone] Fax:[phone]
Fax:[phone] Website:http://www epa.gov/aboutepa/epa-reqion-9-pacific-southeesr
Website:http//www epa gov/aboutepa/abeut.epa-requn-4-southeast Covered states:Arizona,California,Hawaii,Nevada,Guam,American Samoa,
Covered states:Alabama,Florida,Georgia,Kentucky,Mississippi,North Carolina, and Trust Territories
South Carolina,and Tennessee
REGION 5 REGION 10
U.S.Environmental Protection Agency,Region 5 U.S.Environmental Protection Agency,Region 10
77 West Jackson Boulevard,Chicago,IL 60604-3507 1200 Sixth Avenue,Suite 900,Seattle,WA 98101
Plane:[phone];toll free:[phone] Phone:[phone];toll free:[phone]
Fax [phone] Fax:[phone]
Website:http//www epa goviaboutepa/epa-region-5 Website:http://www.epa oov/aboutepa/epa-region-10-pacific-northwest
Covered states:Illinois,Indiana,Michigan,Minnesota,Ohio,and Wisconsin Covered states:Alaska,Idaho,Oregon,and Washington
Exhibit 1-2.Filing Dates for NPDES Permit Applications
Permit Application When to File
180 days before your present NPDES permit expires or,if you are a
2A new discharger,180 days before the date on which the discharge is
to commence unless the NPDES permitting authority has granted
permission for a later date.
2B 180 days before your present NPDES permit expires or 180 days
prior to startup if you are a new facility.
2C 180 days before your present NPDES permit expires.
2D 180 days prior to startup.
2E 180 days before your present NPDES permit expires,or 180 days
prior to startup if you are a new facility_
Construction:90 days prior to date construction is to commence.
2F - Nonconstruction:180 days before your present NPDES permit
expires or 180 days prior to startup if you are a new facility.
2S 180 days before your present NPDES permit expires or 180 days
prior to startup if you are a new facility.
1-3

FORM 1—LINE-BY-LINE INSTRUCTIONS
Section 1.Activities Requiring an NPDES Permit You can find SIC code numbers and descriptions in the
Item 1.1.Review the questions in Item 1.1 to determine if you are 1987 Standard Industrial Classification Manual,prepared by the
required to submit Form 1. Be sure to check the Form 1— Executive Office of the President,Office of Management and
Glossary for the legal definitions of any key terms. Budget This document is available from the Government Printing
Office,Washington, D.C.An online version of the manual is also
If you answer"Yes"to a question in Item 1.1,then you do not available courtesy of the Occupational Safety and Health
need to complete Form 1,but you must comply with the Administration at http://www.osha.govlpls/imis/sic manual.html.
application requirements specified.
You can find NAICS code numbers and descriptions in the North
Item 1.2.Respond to the questions in Items 1.2.1 to 1.2.5.If you American Industrial Classification System Manual prepared by the
answer"Yes"to any question,you must complete Form 1 and the Executive Office of the President,Office of Management and
Form 2 application specified.See Exhibit 1-2 for filing deadlines. Budget.This document is available from the National Technical
If you answer"No"to every question in Items 1.1 and 1.2,then Information Service(NTIS)in Alexandria,Virginia.It is also
you do not need an NPDES permit,and you do not need to available online at http://www census.gov/eos/www/naics/.
complete and return any of the NPDES application forms. Use the latest edition of the manuals.If you have any questions
Section 2.Name,Mailing Address,and Location about the appropriate SIC or NAICS codes for your facility.
contact your NPDES permitting authority.
Item 2.1. Enter the facility's official or legal name.Do not use a
colloquial name. Section 4.Operator Information
Item 2.2.Provide your EPA Identification Number from the Item 4.1.Give the legal name of the person.firm.public
Facility Registry Service if you have an existing facility. If you do organization,or other entity that operates the facility described in
not know your EPA Identification Number,contact your NPDES this application.This may or may not be the same as the facility's
permitting authority.If your facility is new(i.e.,not yet name.The operator of the facility is the legal entity that controls
constructed),write or type"New Facility." the facility's operation rather than the plant or site manager.Do
not use a colloquial name.
Item 2.3.Give the name(first and last),title,work telephone
number,and email address of the person who is thoroughly Item 4.2.Indicate whether the entity listed in response to Item 4.1
familiar with the operation of the facility and with the facts also owns the facility by marking the appropriate box.
reported in this application.The NPDES permitting authority will Item 4.3.Indicate the ownership status of the operator of the
contact the person listed if they have questions on the material facility by marking the appropriate box.If the facility is a federal
submitted. facility(i.e.,owned by the U.S.government),check the box for
Item 2.4.Give the complete mailing address of the office to which "Public—federal."If the facility is owned by a state government,
the NPDES permitting authority should send correspondence. check the box for`Public—state."If the facility is owned by a
This often is not the address used to designate the location of the county government,municipal(e.g.,city or town)government,
facility or activity. tribal government,school district,water district,or other local
government entity,check the box for"Other public"and specify
Item 2.5.Give the address or location of the facility identified the type of government entity. If the facility is owned by a
under Item 2.1. If the facility lacks a street name or route number, corporation or other private entity.check the box for"Private."If
give the most accurate.alternative geographic information(e.g., the facility has mixed ownership(e.g..pubic/private)or is not
section number or quarter section number from county records or owned by an entity of the types previously listed,check the box
"at intersection of Routes 425 and 22').Also provide the county for"Other and specify the type of entity.
name,county code(if known),city or town,state,and zip code.
Items 4.4 to 4.6.Enter the telephone number,address,and email
For concentrated aquatic animal production facilities,provide the address of the operator identified in Item 4.1.
address or location of the production area(i.e.,the location where
the animals are contained,grown,or held). Section 5.Indian Land
Item 5.1.Indicate whether the facility is located on Indian Land.
Section 3.SIC and NAICS Codes
Items 3.1 and 3.2.List,in descending order of significance,up to Section 6.Existing Environmental Permits
four 4-digit standard industrial classification(SIC)codes and Item 6.1.Check the appropriate boxes and provide the permit
North American Industrial Classification System(NAICS)codes numbers for all relevant federal,state,and local environmental
that best describe your facility in terms of the principal products or permits or construction approvals received or applied for under
services it produces or provides.If the SIC or NAICS codes do any of the programs listed below.If you have more than one
not adequately describe your facility's products or services,you currently effective permit under a particular permit program for
have the option to provide additional descriptive information. your facility, list the additional permit numbers on the application
form or on a separate sheet of paper.
1-4

FORM 1—LINE-BY-LINE INSTRUCTIONS CONTINUED
• Hazardous waste management program under the Resource (e.g.,residential,commercial).An example of an acceptable
Conservation and Recovery Act(RCRA). location map is shown as Exhibit 1-3 at the end of these
• Underground Injection Control(UIC)program under the Safe instructions.Note: Exhibit 1-3 is provided for illustration only:it
Drinking Water Act(SDWA). does not show an actual facility.
• NPDES program under the Clean Water Act(CWA). If the facility is a concentrated animal feeding operation,you are
Prevention of Significant Deterioration(PSD)program under not required to provide the topographic map required by this
• the Clean Air Act(CAA). section of Form 1.Instead,you are required to provide a
topographic map as specified in Section 4 of Form 2B.
• Nonattainment program under the CAA.
Item 7.1.Note that you have completed your topographic map
• National Emission Standards for Hazardous Pollutants and attached it to the application.
(NESHAPs)preconstruction approval under the CAA.
Section 8.Nature of Business
• Ocean dumping permits under the Marine Protection
Research and Sanctuaries Act(MPRSA). Briefly describe the nature of your business(e.g.,products
produced or services provided).See Examples 1 and 2.
• Dredge or fill permits under Section 404 of the CWA.
Other federal,state,or local environmental permits. Example 1
• Facilities Subject to 40 CFR 426,Subparts F and G
Section 7.Map Industry A is an auto tempered and auto laminated glass
Unless the facility is a concentrated animal feeding operation, manufacturing facility subject to effluent limitation guidelines
provide a topographic map(s)of the area extending at least one (ELGs)for the"Automotive Glass Tempering"and"Automotive
mile beyond the property boundaries of the facility that clearly Glass Laminating'subcategories of the"Glass Manufacturing"
shows the following: point source category at 40 CFR 426,subparts F and G.At the
The legal boundaries of the facility. facility,glass is cut and then passed through a series of
• processes that grind and polish the edges,bend the glass.and
• The location and serial number of each of your existing and then temper the glass to produce side and back windows for
proposed intake and discharge structures. automobiles.Tempering involves heating the glass near the
• All hazardous waste management,storage,and disposal melting point,then rapidly cooling it to increase its mechanical
facilities. and thermal endurance.The facility also produces automobile
• Each well where you inject fluids underground.
windshields and undertakes processes that laminate a plastic
sheet between two layers of glass and that prepare the glass for
• All wells,springs,surface water bodies,and drinking water lamination(e.g.,cutting,bending,and washing).
wells that are in the public record or otherwise known to you
and that are located in the map area. Example 2
Facility Not Subject to ELGs
If the facility has associated water intakes,discharge structures,
hazardous waste disposal sites,or injection wells and these items Industry B undertakes batch-type resin manufacturing operations.
are located more than one mile from the facility,include them on It has aboveground storage tanks for raw materials and finished
the map if possible.If you cannot.attach additional sheets goods,resin loading operations.and warehouses for 55-gallon
describing the location of the structures,disposal site(s).or well(s) drums of finished product.Industry B manufactures alkyd,
and identify the U.S.Geological Survey(USGS)or other map saturated and unsaturated polyester resins in batches using
corresponding to the location(s). reactor vessels and mix tanks. Most of the feedstock liquids are
pumped from storage tanks to the kettles and mixers via a closed
On each map,include the map scale,a meridian arrow showing piping system.Additional feedstocks are added manually as
north,and latitude and longitude to the nearest second.Latitude solids from bags and sacks via manways,which are located on
and longitude coordinates may be obtained in a variety of ways, top of the kettles.The resin is then chemically reacted in the
including use of hand held devices(e.g.,a GPS enabled kettles.After the reaction step finishes.the resin is transferred
smartphone),Internet mapping tools(e.g., from the kettles to the mix tanks,where solvents are added to
https:!/mynasadata.larc.nasagovilatitudelongitude-finder/), thin it.The primary byproduct of the reaction is water vapor
or
geographic information systems(a•g•,ArcView)�or paper maps
containing condensed soluble organics.The byproduct flows to
from trusted sources(e.g.,USGS). an isolation tank where the vapors are directed to an onsite
On all maps of rivers,show the direction of the current.In tidal thermal oxidizer.The finished resin is then pumped through one
waters.show the directions of ebb and flow tides. of three types of filtration systems into finished goods storage
tanks,55-gallon drums,350-gallon intermediate bulk container
You may develop your map by going to USGS's National Map totes,or directly into tanker trucks.A typical batch takes about 30
website at http://nationalmap.qov/.(For a map from this site,use hours to complete.
the traditional 7.5-minute quadrangle format. If none is available,
use a USGS 15-minute series map.)You may also use a plat or
other appropriate map.Briefly describe land uses in the map area
1-5

FORM 1—LINE•BY-LINE INSTRUCTIONS CONTINUED
Section 9.Cooling Water Intake Structures FEDERAL REGULATIONS AT 40 CFR 122.22 REQUIRE THIS
Item 9.1.Indicate whether the facility uses cooling water.If yes, APPLICATION TO BE SIGNED AS FOLLOWS:
continue to Item 9.2.If no,skip to Item 10.1. A. For a corporation,by a responsible corporate officer.For the
Identify the source of the coolingwater. For example. purpose of this section,a responsible corporate officer
Item 9.2. p
indicate whether the cooling water is from a surface water, means:(1)a president.secretary,treasurer,or vice-president
groundwater well.public water system,or treated effluent that of the corporation in charge of a principal business function,
would otherwise be discharged to a water of the U.S. or any other person who performs similar policy-or decision-
making functions for the corporation.or(2)the manager of
If the facility uses a cooling water intake structure as described in one or more manufacturing,production,or operating facilities.
40 CFR 125,Subparts I and J,the facility may have additional provided the manager is authorized to make management
application requirements under 40 CFR 122.21(r). Note that the decisions which govern the operation of the regulated facility
information required by 40 CFR 122.21(r)is not requested as part including having the explicit or implicit duty of making major
of Form 1.Contact your NPDES permitting authority to determine capital investment recommendations,and initiating and
the specifics of what you should provide and when. directing other comprehensive measures to assure long term
Section 10.Variance Requests environmental compliance with environmental laws and
An applicant(other than a POTW)may request a variance from regulations.the manager can ensure that the necessary
otherwise applicable effluent limitations under certain conditions systems are established or actions taken to gather complete
described at 40 CFR 122.21(m). and accurate information for permit application requirements:
and where authority to sign documents has been assigned or
Item 10.1.If known at the time of application,check all of the delegated to the manager in accordance with corporate
authorized variances that you plan to request or renew.Note that procedures.
you are not being asked to submit any other information at this B. For a partnership or sole proprietorship,by a general partner
time.Contact your NPDES permitting authority to determine the or the proprietor,respectively.
specifics of what you should provide and when.The ability to
request a variance is not limited to the time of application,and an C. For a municipality,state,federal,or other public facility,by
applicant may request a variance consistent with statutory and either a principal executive officer or ranking elected official.
regulatory requirements. For purposes of this section,a principal executive officer of a
federal agency includes:(1)The chief executive officer of the
Section 11.Checklist and Certification agency.or(2)a senior executive officer having responsibility
Item 11.1.Review the checklist provided.In Column 1.mark the for the overall operations of a principal geographic unit of the
sections of Form 1 that you have completed and are submitting agency(e.g.,Regional Administrators of EPA).
with your application.In Column 2,indicate for each section
whether you are submitting attachments.
Item 11.2.The Clean Water Act provides for severe penalties for
submitting false information on this application form.CWA Section
309(c)(2)provides that,"Any person who knowingly makes any
false statement,representation,or certification in any application,
...shall upon conviction.be punished by a fine of no more than
$10,000 or by imprisonment for not more than six months,or
both."
1-6

Exhibit 1-3.Example Topographic Map
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1-7
l

FORM 1—ACTIVITIES THAT DO NOT REQUIRE PERMITS
You are not required to obtain an NPDES permit if your discharge • Any discharge in compliance with the instructions of an On-
is in one of the following categories,as provided by the CWA and Scene Coordinator pursuant to 40 CFR 300(The National Oil
NPDES regulations at 40 CFR 122 to 125.(However,under CWA and Hazardous Substances Pollution Contingency Plan)or
Sections 510 and 312,some discharges exempted from the 33 CFR 153.10(e)(Pollution by Oil and Hazardous
federal NPDES requirements may still be regulated by a state Substances).
permitting authority.) • Any introduction of pollutants from non point-source
• Any discharge of sewage from vessels and any effluent from agricultural and silvicuftural activities,including stormwater
properly functioning marine engines,laundry,shower.and runoff from orchards,cultivated crops,pastures.range lands.
galley sink wastes,or any other discharge incidental to the and forest lands,but not discharges from concentrated
normal operation of a vessel,including vessels of the Armed animal feeding operations as defined in 40 CFR 122.23,
Forces within the meaning of section 312 of the CWA and discharges from concentrated aquatic animal production
recreational vessels within the meaning of section 502(25)of facilities as defined in 40 CFR 122.23,discharges from
the CWA.None of these exclusions apply to rubbish,trash, concentrated aquatic animal production facilities as defined
garbage,or other such materials discharged overboard:nor in 40 CFR 122.24,discharges to aquaculture projects as
to other discharges when the vessel is operating in a defined in 40 CFR 122.25.and discharges from silvicultural
capacity other than as a means of transportation such as point sources as defined in 40 CFR 122.27.Note:Per 40
when used as an energy or mining facility,a storage facility CFR 122.26(b)(14)(ii),facilities classified within SIC 24,
or a seafood processing facility,or when secured to a Industry Group 241.that are rock crushing.gravel washing,
storage facility or a seafood processing facility,or when log sorting,or log storage facilities operated in connection
secured to the bed of the ocean,contiguous zone or waters with silvicultural activities defined in 40 CFR 122.27(b)(2)-(3)
of the United States for the purpose of mineral or oil and Industry Groups 242 through 249:26(except 265 and
exploration or development. 267),28(except 283),29,311,32(except 323),33,3441.
and 373(not included are all other types of silviculture
• Discharges of dredged or fill material into waters of the facilities)are considered stormwater discharges associated
United States that are regulated under CWA Section 404. with industrial activity,and are required to obtain an NPDES
• The introduction of sewage,industrial wastes,or other permit.
pollutants into publicly owned treatment works by indirect • Return flows from irrigated agriculture.
dischargers.Plans or agreements to switch to this method of
disposal in the future do not relieve dischargers of the • Discharges into a privately owned treatment works,except as
obligation to have and comply with permits until all the NPDES permitting authority may otherwise require under
discharges of pollutants to waters of the United States are 40 CFR 122.44(m).
eliminated. (See also 40 CFR 122.47(b).)This exclusion • Discharges from a water transfer.'Water transfer"means an
does not apply to the introduction of pollutants to privately activity that conveys or connects waters of the United States
owned treatment works or to other discharges through pipes, without subjecting the transferred water to intervening
sewers,or other conveyances owned by a state,municipality, industrial,municipal,or commercial use.This exclusion does
or other party not leading to treatment works. not apply to pollutants introduced by the water transfer
activity itself to the water being transferred.
1-8

FORM 1—GLOSSARY
Note:This glossary includes terms used in the various NPDES application forms,including Form 1.The definitions are from the NPDES
regulations at 40 CFR 122.2 unless otherwise specified.If you have any questions concerning the meaning of any of these terms,contact
your NPDES permitting authority.
ANIMAL FEEDING OPERATION(defined at§ 122.23)means a lot or facility(other than an aquatic animal production facility)where the
following conditions are met;
• Animals(other than aquatic animals)have been, are,or will be stabled or confined and fed or maintained for a total of 45 days or
more in any 12-month period;and
• Crops,vegetation,forage growth,or post-harvest residues are not sustained in the normal growing season over any portion of the lot
or facility.
APPLICATION means the EPA standard national forms for applying for a permit.including any additions,revisions,or modifications to the
forms;or forms approved by EPA for use in approved states,including any approved modifications or revisions.
APPROVED PROGRAM or APPROVED STATE means a State or interstate program which has been approved or authorized by EPA
under part 123.
AQUACULTURE PROJECT(defined at§122.25)means a defined managed water area which uses discharges of pollutants into that
designated area for the maintenance or production of harvestable freshwater,estuarine,or marine plants or animals.DESIGNATED
PROJECT AREA means the portions of the waters of the United States within which the permittee or permit applicant plans to confine the
cultivated species,using a method or plan or operation(including,but not limited to,physical confinement)which,on the basis of reliable
scientific evidence.is expected to ensure that specific individual organisms comprising an aquaculture crop will enjoy increased growth
attributable to the discharge of pollutants,and be harvested within a defined geographic area.
AVERAGE MONTHLY DISCHARGE LIMITATION means the highest allowable average of daily discharges over a calendar month,
calculated as the sum of all daily discharges measured during that month divided by the number of daily discharges measured during that
month.
AVERAGE WEEKLY DISCHARGE LIMITATION means the highest allowable average of daily discharges over a calendar week,
calculated as the sum of all daily discharges measured during a calendar week divided by the number of daily discharges measured during
that week.
BEST MANAGEMENT PRACTICES(BMPs)means schedules of activities,prohibitions of practices,maintenance procedures,and other
management practices to prevent or reduce the pollution of waters of the United States. BMPs include treatment requirements,operation
plant site runoff,spillage or leaks,sludge or waste disposal,or drainage from raw material storage.
procedures,and practices to controlg sp g g
BIOSOLIDS(see sewage sludge).
BYPASS(defined at§122.41(m))means the intentional diversion of waste streams from any portion of a treatment facility.
COMBINED SEWER OVERFLOW(CSO)means a discharge from a combined sewer system(CSS)at a point prior to the Publicly Owned
Treatment Works(POTW)Treatment Plant(defined at§403.3(r)).
COMBINED SEWER SYSTEM(CSS)means a wastewater collection system owned by a State or municipality(as defined by section
502(4)of the CWA)which conveys sanitary wastewaters(domestic,commercial and industrial wastewaters)and storm water through a
single-pipe system to a Publicly Owned Treatment Works(POTW)Treatment Plant(as defined at§403.3(r)).
CONCENTRATED ANIMAL FEEDING OPERATION(defined at§122.23)means an animal feeding operation that is defined as a Large
CAFO or as a Medium CAFO by the terms of(A)or(B)below,or that is designated as a CAFO in accordance with 40 CFR 122.23(c).Two
or more AFOs under common ownership are considered to be a single AFO for the purposes of determining the number of animals at an
operation,if they adjoin each other or if they use a common area or system for the disposal of wastes.
A. LARGE CONCENTRATED ANIMAL FEEDING OPERATION(LARGE CAFO)means an AFO that stables or confines as many as or
more than the numbers of animals specified in any of the following categories:
1. 700 mature dairy cows.whether milked or dry;
2. 1,000 veal calves;
3. 1,000 cattle other than mature dairy cows or veal calves.Cattle includes but is not limited to heifers.steers,bulls and cow/calf
pairs:
4. 2,500 swine each weighing 55 pounds or more;
5. 10,000 swine each weighing less than 55 pounds;
1-9

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4

FORM 1—GLOSSARY CONTINUED
6. 500 horses;
7. 10.000sheep
or lambs:
8. 55,000 turkeys;
9. 30,000 laying hens or broilers,if the AFO uses a liquid manure handling system;
10. 125,000 chickens(other than laying hens),if the AFO uses other than a liquid manure handling system;
11. 82,000 laying hens,if the AFO uses other than a liquid manure handling system;
12. 30,000 ducks(if the AFO uses other than a liquid manure handling system);or
13. 5,000 ducks(if the AFO uses a liquid manure handling system).
B. MEDIUM CONCENTRATED ANIMAL FEEDING OPERATION(MEDIUM CAFO)means any AFO with the type and number of
animals that fall within any of the ranges listed below and which has been defined or designated as a CAFO.An AFO is defined as a
Medium CAFO if:
1. The type and number of animals that it stables and confines falls within any of the following ranges:
a. 200 to 699 mature dairy cows,whether milked or dry;
b. 300 to 999 veal calves:
c. 300 to 999 cattle other than mature dairy cows or veal calves.Cattle includes but is not limited to heifers,steers,bulls and
cow/calf pairs:
d. 750 to 2,499 swine each weighing 55 pounds or more;
e. 3,000 to 9,999 swine each weighing less than 55 pounds;
f. 150 to 499 horses:
g. 3,000 to 9,999 sheep or lambs;
h. 16,500 to 54,999 turkeys;
layinghens or broilers.if the AFO uses a liquid manure handlingsystem;
I. 9,000 to 29,999q Y
j. 37,500 to 124,999 thickens(other than laying hens),if the AFO uses other than a liquid manure handling system;
k. 25,000 to 81,999 laying hens,if the AFO uses other than a liquid manure handling system;
I. 10,000 to 29.999 ducks(if the AFO uses other than a liquid manure handling system);ore
m. 1,500 to 4,999 ducks(if the AFO uses a liquid manure handling system):and
2. Either one of the following conditions are met:
a. Pollutants are discharged into waters of the United States through a man-made ditch,flushing system.or other similar man-
made device;or
b. Pollutants are discharged directly into waters of the United States which originate outside of and pass over,across,or
through the facility or otherwise come into direct contact with animals confined in the operation.
CONCENTRATED AQUATIC ANIMAL PRODUCTION FACILITY(defined at§122.24)means a hatchery,fish farm,or other facility which
contains,grows,or holds aquatic animals in either of the following categories,or which the Director designates as such on a case-by-case
basis:
A. Cold water fish species or other cold water aquatic animals including,but not limited to,the Salmonidae family of fish(e.g.,trout and
salmon)in ponds.raceways,or other similar structures which discharge at least 30 days per year but does not include:
1. Facilities which produce less than 9,090 harvest weight kilograms(approximately 20,000 pounds)of aquatic animals per year;
and
2. Facilities which feed less than 2,272 kilograms(approximately 5,000 pounds)of food during the calendar month of maximum
feeding.
B. Warm water fish species or other warm water aquatic animals including,but not limited to,the Ameiuridae, Cetrarchiclae,and
Cyprinidae families of fish(e.g.,respectively,catfish,sunfish,and minnows)in ponds,raceways,or other similar structures which
discharge at least 30 days per year,but does not include:
1. Closed ponds which discharge only during periods of excess runoff;or
2. Facilities which produce less than 45,454 harvest weight kilograms(approximately 100,000 pounds)of aquatic animals per year.
1-10

FORM 1—GLOSSARY CONTINUED
CWA means the Clean Water Act(formerly referred to as the Federal Water Pollution Control Act or Federal Water Pollution Control Act
Amendments of 1972)Public Law 92-500,as amended by Public Law 95-217,Public Law 95-576,Public Law 96-483 and Public Law
97-117,33 U.S.C. 1251 et seq.
CWA AND REGULATIONS means the Clean Water Act(CWA)and applicable regulations promulgated thereunder. In the case of an
approved State program,it includes State program requirements.
DAILY DISCHARGE means the"discharge of a pollutant"measured during a calendar day or any 24-hour period that reasonably
represents the calendar day for purposes of sampling.For pollutants with limitations expressed in units of mass,the"daily discharge"is
calculated as the total mass of the pollutant discharged over the day. For pollutants with limitations expressed in other units of
measurement,the"daily discharge"is calculated as the average measurement of the pollutant over the day.
DIRECT DISCHARGE means the"discharge of a pollutant"
DIRECTOR means the Regional Administrator or the State Director,as the context requires,or an authorized representative.When there
is no"approved State program,'and there is an EPA administered program,"Director"means the Regional Administrator.When there is
an approved State program,"Director'normally means the State Director.In some circumstances.however,EPA retains the authority to
take certain actions even when there is an approved State program. (For example,when EPA has issued an NPDES permit prior to the
approval of a State program, EPA may retain jurisdiction over that permit after program approval,see§123.1.)In such cases.the term
"Director"means the Regional Administrator and not the State Director.
DISCHARGE(OF A POLLUTANT)means:
• Any addition of any pollutant or combination of pollutants to waters of the United States from any point source:or
• Any addition of any pollutant or combination of pollutants to the waters of the contiguous zone or the ocean from any point source
other than a vessel or other floating craft which is being used as a means of transportation.
This definition includes discharges into waters of the United States from:surface runoff which is collected or channelled by man;
discharges through pipes,sewers,or other conveyances owned by a State,municipality,or other person which do not lead to a treatment
works:and discharges through pipes.sewers.or other conveyances,leading into privately owned treatment works.This term does not
include an addition of pollutants by any"indirect discharger.
DISCHARGE MONITORING REPORT means the EPA uniform national form,including any subsequent additions.revisions,or
modifications for the reporting of self-monitoring results by permittees. DMRs must be used by"approved States"as well as by EPA. EPA
will supply DMRs to any approved State upon request.The EPA national forms may be modified to substitute the state agency name,
address,logo,and other similar information,as appropriate,in place of EPA's.
DRAFT PERMIT means a document prepared under§ 124.6 indicating the Director's tentative decision to issue or deny,modify,revoke
and reissue,terminate.or reissue a"permit."A notice of intent to terminate a permit, and a notice of intent to deny a permit,as discussed
in§124.5,are types of"draft permits."A denial of a request for modification,revocation and reissuance,or termination,as discussed in
§ 124.5,is not a"draft permit."A"proposed permit"is not a"draft permit"
EFFLUENT LIMITATION means any restriction imposed by the Director on quantities,discharge rates, and concentrations of"pollutants"
which are"discharged'from"point sources"into"waters of the United States,"the waters of the"contiguous zone."or the ocean.
EFFLUENT LIMITATIONS GUIDELINES means a regulation published by the Administrator under section 304(b)of the CWA to adopt or
revise"effluent limitations."
ENVIRONMENTAL PROTECTION AGENCY(EPA)means the United States Environmental Protection Agency.
FACILITY or ACTIVITY means any NPDES"point source"or any other facility or activity(including land or appurtenances thereto)that is
subject to regulation under the NPDES program.
GENERAL PERMIT means an NPDES"permit"issued under§122.28 authorizing a category of discharges under the CWA within a
geographical area.
HAZARDOUS SUBSTANCE means any substance designated under 40 CFR part 116 pursuant to section 311 of the CWA.
INDIAN COUNTRY(or INDAN LANDS)means:
• All land within the limits of any Indian reservation under the jurisdiction of the United States Government,notwithstanding the
issuance of any patent,and,including rights-of-way running through the reservation;
• All dependent Indian communities with the borders of the United States whether within the originally or subsequently acquired territory
thereof,and whether within or without the limits of a state;and
• All Indian allotments,the Indian titles to which have not been extinguished,including rights-of-way running through the same.
1-11

FORM 1—GLOSSARY CONTINUED
INDIAN TRIBE means any Indian Tribe,band,group,or community recognized by the Secretary of the Interior and exercising
governmental authority over a Federal Indian reservation.
INDIRECT DISCHARGE means a nondomestic discharger introducing"pollutants"to a"publicly owned treatment works."
LARGE MUNICIPAL SEPARATE STORM SEWER SYSTEM(defined at§122.26(b)(4))means all municipal separate storm sewers that
are either:
(i)Located in an incorporated place with a population of 250,000 or more as determined by the 1990 Decennial Census by the Bureau of
the Census(Appendix F of 40 CFR 122);or
(ii)Located in the counties listed in appendix H of 40 CFR 122,except municipal separate storm sewers that are located in the
incorporated places,townships or towns within such counties;or
(iii)Owned or operated by a municipality other than those described in paragraphs(i)or(ii)and that are designated by the Director as part
of the large or medium municipal separate storm sewer system due to the interrelationship between the discharges of the designated
storm sewer and the discharges from municipal separate storm sewers described under paragraphs(i)or(ii).In making this determination
the Director may consider the following factors:
(A)Physical interconnections between the municipal separate storm sewers;
(B)The location of discharges from the designated municipal separate storm sewer relative to discharges from municipal separate storm
sewers described in paragraph(i);
(C)The quantity and nature of pollutants discharged to waters of the United States:
(D)The nature of the receiving waters;and
(E)Other relevant factors;or
(iv)The Director may,upon petition,designate as a large municipal separate storm sewer system,municipal separate storm sewers
located within the boundaries of a region defined by a storm water management regional authority based on a jurisdictional.watershed.or
other appropriate basis that includes one or more of the systems described in paragraphs(i),(ii),(iii).
LOG SORTING AND LOG STORAGE FACILITIES(defined at§122.27)means facilities whose discharges result from the holding of
unprocessed wood,for example,logs or roundwood with bark or after removal of bark held in self-contained bodies of water(mill ponds or
log ponds)or stored on land where water is applied intentionally on the logs(wet decking).(See 40 CFR 429,subpart I,including the
effluent limitations guidelines.)
MAJOR FACILITY means any NPDES"facility or activity"classified as such by the Regional Administrator,or.in the case of"approved
State programs,"the Regional Administrator in conjunction with the State Director.
MAXIMUM DAILY DISCHARGE LIMITATION means the highest allowable"daily discharge."
MEDIUM MUNICIPAL SEPARATE STORM SEWER SYSTEM(defined at§122.26(b)(7))means all municipal separate storm sewers that
are either:
(i)Located in an incorporated place with a population of 100,000 or more but less than 250,000,as determined by the 1990 Decennial
Census by the Bureau of the Census(appendix G of 40 CFR 122);or
(ii)Located in the counties listed in appendix I of 40 CFR 122,except municipal separate storm sewers that are located in the incorporated
places,townships or towns within such counties:or
(iii)Owned or operated by a municipality other than those described in paragraph(i)or(ii)and that are designated by the Director as part
of the large or medium municipal separate storm sewer system due to the interrelationship between the discharges of the designated
storm sewer and the discharges from municipal separate storm sewers described under paragraph(i)or(ii).In making this determination
the Director may consider the following factors:
(A)Physical interconnections between the municipal separate storm sewers:
(B)The location of discharges from the designated municipal separate storm sewer relative to discharges from municipal separate storm
sewers described in paragraph(i);
(C)The quantity and nature of pollutants discharged to waters of the United States;
(D)The nature of the receiving waters;or
(E)Other relevant factors;or
1-12

FORM 1—GLOSSARY CONTINUED
(iv)The Director may,upon petition,designate as a medium municipal separate storm sewer system,municipal separate storm sewers
located within the boundaries of a region defined by a storm water management regional authority based on a jurisdictional,watershed,or
other appropriate basis that includes one or more of the systems described in paragraphs(i),(ii),(Hi)of this section.
MUNICIPALITY means a city,town,borough,county,parish,district,association,or other public body created by or under State law and
having jurisdiction over disposal of sewage,industrial wastes,or other wastes,or an Indian tribe or an authorized Indian tribal organization,
or a designated and approved management agency under section 208 of the CWA.
MUNICIPAL SEPARATE STORM SEWER(defined at§ 122.26(b)(8))means a conveyance or system of conveyances(including roads
with drainage systems,municipal streets,catch basins,curbs,gutters,ditches,man-made channels,or storm drains):
• Owned or operated by a State,city,town,borough.county,parish,district,association,or other public body(created by or pursuant to
State law)having jurisdiction over disposal of sewage,industrial wastes,stormwater,or other wastes,including special districts under
State law such as a sewer district.flood control district or drainage district,or similar entity,or an Indian tribe or an authorized Indian
tribal organization,or a designated and approved management agency under section 208 of the CWA that discharges to waters of the
United States.
• Designed or used for collecting or conveying stormwater.
• Which is not a combined sewer; and
• Which is not part of a POTW as defined at 40 CFR 122.2.
MUNICIPAL SLUDGE(see sewage sludge)
NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM(NPDES)means the national program for issuing,modifying,revoking
and reissuing,terminating,monitoring and enforcing permits.and imposing and enforcing pretreatment requirements,under sections 307,
402,318,and 405 of the CWA.The term includes an"approved program."
NEW DISCHARGER means any building.structure,facility,or installation:
• From which there is or may be a"discharge of pollutants;"
• That did not commence the"discharge of pollutants"at a particular"site"prior to August 13, 1979;
• Which is not a"new source;"and
• Which has never received a finally effective NPDES permit for discharges at that"site."
This definition includes an"indirect discharger"which commences discharging into"waters of the United States"after August 13, 1979. It
also means any existing mobile point source(other than an offshore or coastal oil and gas exploratory drilling rig or a coastal oil and gas
developmental drilling rig)such as a seafood processing rig,seafood processing vessel,or aggregate plant,that begins discharging at a
"site"for which it does not have a permit;and any offshore or coastal mobile oil and gas exploratory drilling rig or coastal mobile oil and gas
developmental drilling rig that commences the discharge of pollutants after August 13. 1979.at a"site"under EPA's permitting jurisdiction
for which it is not covered by an individual or general permit and which is located in an area determined by the Regional Administrator in
the issuance of a final permit to be an area of biological concern.In determining whether an area is an area of biological concern.the
Regional Administrator shall consider the factors specified in 40 CFR 125.122(a)(1)through(10).
An offshore or coastal mobile exploratory drilling rig or coastal mobile developmental drilling rig will be considered a"new discharger"only
for the duration of its discharge in an area of biological concern.
NEW SOURCE means any building,structure,facility,or installation from which there is or may be a"discharge of pollutants,"the
construction of which commenced:
• After promulgation of standards of performance under section 306 of the CWA which are applicable to such source,or
• After proposal of standards of performance in accordance with section 306 of the CWA which are applicable to such source.but only
if the standards are promulgated in accordance with section 306 within 120 days of their proposal.
OWNER OR OPERATOR means the owner or operator of any`facility or activity"subject to regulation under the NPDES program.
PERMIT means an authorization, license,or equivalent control document issued by EPA or an"approved State"to implement the
requirements of this part and parts 123 and 124."Permit"includes an NPDES"general permit"(§ 122.28).Permit does not include any
permit which has not yet been the subject of final agency action,such as a"draft permit"or a"proposed permit"
PESTICIDE DISCHARGES TO WATERS OF THE UNITED STATES FROM PESTICIDE APPLICATION means the application of
biological pesticides,and the application of chemical pesticides that leave a residue,from point sources to waters of the United States. In
the context of this definition of pesticide discharges to waters of the United States from pesticide application,this does not include
1-13

FORM 1—GLOSSARY CONTINUED
agricultural storm water discharges and return flows from irrigated agriculture,which are excluded by law(33 U.S.C. 1342(1);33 U.S.C.
1362(14)).
PESTICIDE RESIDUE for the purpose of determining whether a NPDES permit is needed for discharges to waters of the United States
frompesticide application.means thatportion of apesticide application that is discharged from a point source to waters of the United
PP PP 9
States and no longer provides pesticidal benefits. It also includes any degradates of the pesticide.
POINT SOURCE means any discernible,confined,and discrete conveyance,including but not limited to,any pipe,ditch,channel,tunnel,
conduit,well,discrete fissure,container,rolling stock,concentrated animal feeding operation,landfill leachate collection system,vessel or
other floating craft from which pollutants are or may be discharged.This term does not include return flows from irrigated agriculture or
agricultural stormwater runoff.(See§122.3).
POLLUTANT means dredged spoil,solid waste,incinerator residue,filter backwash,sewage,garbage,sewage sludge,munitions.
chemical wastes.biological materials.radioactive materials(except those regulated under the Atomic Energy Act of 1954,as amended(42
U.S.C.2011 et seq.)),heat,wrecked or discarded equipment,rock,sand,cellar dirt and industrial,municipal,and agricultural waste
discharged into water.It does not mean:
• Sewage from vessels;or
• Water,gas,or other material which is injected into a well to facilitate production of oil or gas,or water derived in association with oil
and gas production and disposed of in a well.if the well used either to facilitate production or for disposal purposes is approved by
authority of the State in which the well is located,and if the State determines that the injection or disposal will not result in the
degradation of ground or surface water resources.Note: Radioactive materials covered by the Atomic Energy Act are those
encompassed in its definition of source,byproduct,or special nuclear materials.Examples of materials not covered include radium
and accelerator-produced isotopes.See Train v.Colorado Public Interest Research Group. Inc.,426 U.S. 1 (1976).
PRIMARY INDUSTRY CATEGORY means any industry category listed in the NRDC settlement agreement(Natural Resources Defense
Council et at.v. Train,8 E.R.C.2120(D.D.C. 1976),modified 12 E.R.C. 1833(D.D.C. 1979));also listed in appendix A of part 122.
PRIVATELY OWNED TREATMENT WORKS means any device or system which is(1)used to treat wastes from any facility whose
operator is not the operator of the treatment works and(2)not a"POTW."
PROCESS WASTEWATER means any water which,during manufacturing or processing,comes into direct contact with or results from the
production or use of any raw material,intermediate product,finished product,byproduct,or waste product.
PROPOSED PERMIT means a state NPDES"permit"prepared after the close of the public comment period(and.when applicable,any
public hearing and administrative appeals)which is sent to EPA for review before final issuance by the State.A"proposed permit"is not a
-draft permit."
PUBLICLY OWNED TREATMENT WORKS or POTW(defined at§403.3)means a treatment works as defined by CWA Section 212,
which is owned by a state or municipality(as defined by CWA Section 502(4)).This definition includes any devices or systems used in the
storage,treatment,recycling,and reclamation)of municipal sewage or industrial wastes of a liquid nature.This definition also includes
sewers,pipes.and other conveyances only if they convey wastewater to a POTW.The term also means the municipality as defined in
CWA Section 502(4),which has jurisdiction over the indirect discharges to and the discharges from such a treatment works.
REGIONAL ADMINISTRATOR means the Regional Administrator of the appropriate Regional Office of the Environmental Protection
Agency or the authorized representative of the Regional Administrator.
ROCK CRUSHING AND GRAVEL WASHING FACILITIES(defined at§122.27)means facilities which process crushed and broken
stone,gravel,and riprap(See 40 CFR 436,subpart B,including the effluent limitations guidelines).
SCHEDULE OF COMPLIANCE means a schedule of remedial measures included in a"permit',including an enforceable sequence of
interim requirements(for example,actions,operations.or milestone events)leading to compliance with the CWA and regulations.
SECONDARY INDUSTRY CATEGORY means any industry category which is not a primary industry category.
SEWAGE FROM VESSELS means human body wastes and the wastes from toilets and other receptacles intended to receive or retain
body wastes that are discharged from vessels and regulated under section 312 of the CWA,except that with respect to commercial
vessels on the Great Lakes this term includes graywater.For the purposes of this definition."graywater"means galley,bath,and shower
water.
SEWAGE SLUDGE means any solid,semi-solid,or liquid residue removed during the treatment of municipal waste water or domestic
sewage.Sewage sludge includes,but is not limited to,solids removed during primary,secondary,or advanced waste water treatment,
scum,septage.portable toilet pumpings,type Ill marine sanitation device pumpings(33 CFR 159),and sewage sludge products. Sewage
sludge does not include grit or screenings,or ash generated during the incineration of sewage sludge.
1-14

FORM 1—GLOSSARY CONTINUED
SILVICULTURAL POINT SOURCE(defined at§122.27)means any discernible,confined,and discrete conveyance related to rock
crushing.gravel washing,log sorting,or log storage facilities which are operated in connection with silvicultural activities and from which
pollutants are discharged into waters of the United States.This term does not include non-point source silvicultural activities such as
nursery operations,site preparation,reforestation and subsequent cultural treatment,thinning,prescribed burning,pest and fire control,
harvesting operations,surface drainage,or road construction and maintenance from which there is natural runoff.However,some of these
activities(such as stream crossing for roads)may involve point source discharges of dredged or fill material which may require a CWA
Section 404 permit(see 33 CFR 209.120 and part 233).
SITE means the land or water area where any"facility or activity"is physically located or conducted,including adjacent land used in
connection with the facility or activity.
SLUDGE-ONLY FACILITY means any"treatment works treating domestic sewage"whose methods of sewage sludge use or disposal are
subject to regulations promulgated pursuant to section 405(d)of the CWA and is required to obtain a permit under§122.1(b)(2).
STANDARDS FOR SEWAGE SLUDGE USE OR DISPOSAL means the regulations promulgated pursuant to section 405(d)of the CWA
which govern minimum requirements for sludge quality,management practices,and monitoring and reporting applicable to sewage sludge
or the use or disposal of sewage sludge by any person.
STATE means any of the 50 States,the District of Columbia.Guam,the Commonwealth of Puerto Rico,the Virgin Islands,American
Samoa,the Commonwealth of the Northern Mariana Islands,the Trust Territory of the Pacific Islands,or an Indian Tribe as defined in
these regulations which meets the requirements of§ 123.31 of this chapter.
STATE DIRECTOR means the chief administrative officer of any State or interstate agency operating an"approved program."or the
delegated representative of the State Director.If responsibility is divided among two or more State or interstate agencies,"State Director"
means the chief administrative officer of the State or interstate agency authorized to perform the particular procedure or function to which
reference is made.
STORMWATER(or STORM WATER)(defined at§122.26(b)(13))means stormwater runoff,snow melt runoff,and surface runoff and
drainage.
STORMWATER DISCHARGE ASSOCIATED WITH INDUSTRIAL ACTIVITY(defined at§122.26(b)(14))means the discharge from any
conveyance that is used for collecting and conveying stormwater and that is directly related to manufacturing,processing or raw materials
storage areas at an industrial plant.The term does not include discharges from facilities or activities excluded from the NPDES program
under this part 122.For the categories of industries identified in this section,the term includes,but is not limited to,stormwater discharges
from industrial plant yards;immediate access roads and rail lines used or traveled by carriers of raw materials.manufactured products.
waste material,or by-products used or created by the facility;material handling sites;refuse sites;sites used for the application or disposal
of process waste waters(as defined at 40 CFR 401);sites used for the storage and maintenance of material handling equipment;sites
used for residual treatment,storage.or disposal;shipping and receiving areas;manufacturing buildings;storage areas(including tank
farms)for raw materials,and intermediate and final products;and areas where industrial activity has taken place in the past and significant
materials remain and are exposed to stormwater. For the purposes of this paragraph,material handling activities include storage,loading
and unloading,transportation,or conveyance of any raw material,intermediate product,final product,by-product or waste product.The
term excludes areas located on plant lands separate from the plant's industrial activities,such as office buildings and accompanying
parking lots as long as the drainage from the excluded areas is not mixed with stormwater drained from the above described areas.
Industrial facilities(including industrial facilities that are federally,State,or municipally owned or operated that meet the description of the
facilities listed in paragraphs 1 through 14 below)include those facilities designated under the provisions of 40 CFR 122.26(a)(1)(v).The
following categories of facilities are considered to be engaging in"industrial activity"for purposes of 40 CFR 122.26(b)(14):
1. Facilities subject to stormwater effluent limitations guidelines,new source performance standards,or toxic pollutant effluent standards
under 40 CFR Subchapter N(except facilities with toxic pollutant effluent standards which are exempted under paragraph 11 below):
2. Facilities classified as Standard Industrial Classification 24, Industry Group 241 that are rock crushing,gravel washing,log sorting.or
log storage facilities operated in connection with silvicultural activities defined in 40 CFR 122.27(b)(2)—(3)and Industry Groups 242
through 249;26(except 265 and 267),28(except 283),29,311,32(except 323),33,3441,373:(not included are all other types of
silvicultural facilities);
3. Facilities classified as Standard Industrial Classifications 10 through 14(mineral industry)including active or inactive mining
operations(except for areas of coal mining operations no longer meeting the definition of a reclamation area under 40 CFR 434.11(1)
because the performance bond issued to the facility by the appropriate SMCRA authority has been released,or except for areas of
non—coal mining operations which have been released from applicable State or Federal reclamation requirements after
December 17, 1990)and oil and gas exploration,production,processing,or treatment operations,or transmission facilities that
discharge stormwater contaminated by contact with or that has come into contact with,any overburden,raw material,intermediate
products,finished products,byproducts or waste products located on the site of such operations;(inactive mining operations are
mining sites that are not being actively mined.but which have an identifiable owner/operator:inactive mining sites do not include sites
1-15

FORM 1—GLOSSARY CONTINUED
where mining claims are being maintained prior to disturbances associated with the extraction,beneficiation,or processing of mined
materials.nor sites where minimal activities are undertaken for the sole purpose of maintaining a mining claim);
4. Hazardous waste treatment, storage,or disposal facilities,including those that are operating under interim status or a permit under
subtitle C of RCRA:
5. Landfills,land application sites,and open dumps that receive or have received any industrial wastes(waste that is received from any
of the facilities described under this subsection)including those that are subject to regulation under subtitle D of RCRA;
6. Facilities involved in the recycling of materials,including metal scrapyards,battery reclaimers,salvage yards,and automobile
junkyards.including but limited to those classified as Standard Industrial Classification 5015 and 5093;
7. Steam electric power generating facilities.including coal handling sites;
8. Transportation facilities classified as Standard Industrial Classifications 40,41,42(except 4221-25),43,44,45,and 5171 which have
vehicle maintenance shops,equipment cleaning operations,or airport deicing operations.Only those portions of the facility that are
either involved in vehicle maintenance(including vehicle rehabilitation,mechanical repairs,painting,fueling,and lubrication),
equipment cleaning operations,airport deicing operations,or which are otherwise identified under paragraphs 1-7 or 9-11 are
associated with industrial activity;
9. Treatment works treating domestic sewage or any other sewage sludge or wastewater treatment device or system,used in the
storage treatment,recycling,and reclamation of municipal or domestic sewage,including land dedicated to the disposal of sewage
sludge that are located within the confines of the facility,with a design flow of 1.0 mgd or more,or required to have an approved
pretreatment program under 40 CFR 403.Not included are farm lands,domestic gardens or lands used for sludge management
where sludge is beneficially reused and which are not physically located in the confines of the facility,or areas that are in compliance
with section 405 of the CWA;
10. Construction activity including clearing,grading and excavation,except operations that result in the disturbance of less than five acres
of total land area.Construction activity also includes the disturbance of less than five acres of total land area that is a part of a larger
common plan of development or sale if the larger common plan will ultimately disturb five acres or more;
11. Facilities under Standard Industrial Classifications 20,21,22.23,2434,25,265,267,27,283,285,30,31 (except 311).323,34
(except 3441).35,36,37(except 373).38,39,and 4221-25.
TOXIC POLLUTANT means any pollutant listed as toxic under section 307(a)(1)or,in the case of"sludge use or disposal practices,"any
pollutant identified in regulations implementing section 405(d)of the CWA.
TREATMENT WORKS TREATING DOMESTIC SEWAGE(TWTDS)means a POTW or any other sewage sludge or waste water
treatment devices or systems,regardless of ownership(including federal facilities),used in the storage,treatment,recycling,and
reclamation of municipal or domestic sewage.including land dedicated for the disposal of sewage sludge.This definition does not include
septic tanks or similar devices.For purposes of this definition,"domestic sewage"includes waste and waste water from humans or
household operations that are discharged to or otherwise enter a treatment works.In States where there is no approved State sludge
management program under section 405(f)of the CWA,the Regional Administrator may designate any person subject to the standards for
sewage sludge use and disposal in 40 CFR 503 as a"treatment works treating domestic sewage,"where he or she finds that there is a
potential for adverse effects on public health and the environment from poor sludge quality or poor sludge handling,use or disposal
practices,or where he or she finds that such designation is necessary to ensure that such person is in compliance with 40 CFR 503.
UPSET(defined at§122.41(n))means an exceptional incident in which there is unintentional and temporary noncompliance with
technology based permit effluent limitations because of factors beyond the reasonable control of the permittee.An upset does not include
noncompliance to the extent caused by operational error,improperly designed treatment facilities, inadequate treatment facilities,lack of
preventive maintenance,or careless or improper operation.
VARIANCE means any mechanism or provision under section 301 or 316 of the CWA or under 40 CFR 125,or in the applicable"effluent
limitations guidelines"which allows modification to or waiver of the generally applicable effluent limitation requirements or time deadlines of
the CWA.This includes provisions which allow the establishment of alternative limitations based on fundamentally different factors or on
sections 301(c),301(g),301(h),301(i),or 316(a)of the CWA.
WATERS OF THE UNITED STATES as defined at§122,2.
WHOLE EFFLUENT TOXICITY(WET)means the aggregate toxic effect of an effluent measured directly by a toxicity test
1-16

EPA Identification Number NPDES Permit Number Facility Name Form Approved 03/05/19
93647 NCG560000 BlueGreen Water Technologies OMB No.2040 0004
Form U.S.Environmental Protection Agency
� 1 pw Application for NPDES Permit to Discharge Wastewater
NPDES G �/`1 GENERAL INFORMATION
SECTION 1.ACTIVITIES REQUIRING AN NPDES PERMIT(40 CFR 122.21(f)and(f)(1))
1.1 Applicants Not Required to Submit Form 1
Is the facility a new or existing publicly owned Is the facility a new or existing treatment works
1.1.1 treatment works? 1.1.2 treating domestic sewage?
If yes,STOP. Do NOT complete No If yes,STOP. Do NOT ID No
Form 1.Complete Form 2A. complete Form 1.Complete
Form 2S.
1.2 Applicants Required to Submit Form 1
1.2.1 Is the facility a concentrated animal feeding 1.2.2 Is the facility an existing manufacturing,
operation or a concentrated aquatic animal commercial,mining,or silvicultural facility that Is
a production facility? currently discharging process wastewater?
EI Yes 4 Complete Form 1 El No Yes 4 Complete Form ✓� No
and Form 26. 1 and Form 2C.
c 1.2.3 Is the facility a new manufacturing,commercial, 1.2.4 Is the facility a new or existing manufacturing,
ta
= mining,or silvicultural facility that has not yet commercial.mining,or silvicultural facility that
: commenced to discharge? discharges only nonprocess wastewater?
g Yes 4 Complete Form 1 ✓� No El Yes 4 Complete Form E] No
cc and Form 2D. 1 and Form 2E.
40
°t 1.2.5 Is the facility a new or existing facility whose
discharge is composed entirely of stormwater
associated with industrial activity or whose
discharge is composed of both stormwater and
non-stormwater?
Yes 4 Complete Form 1 ✓❑ No
and Form 2F
unless exempted by
40 CFR
122.26(b)(14)(x)or
b 15.
SECTION 2. NAME, MAILING ADDRESS,AND LOCATION(40 CFR 122.21(f)(2))
2.1 Facility Name
BlueGreen Water Technologies
2.2 EPA Identification Number
r
O 93647
2.3 Facility Contact
Name(first and last) Title Phone number
v
Jessica Frost Scientific Director [phone]
co Email address
[e-mail]
2.4 Facility Mailing Address
co Street or P.O.box
301 South Hills Village Ste LL200#452
City or town State ZIP code
Pittsburgh PA 15241
EPA Form 3510-1(revised 3-19) Page 1

EPA Identification Number NPDES Permit Number Facility Name Form Approved 03/05/19
93647 NCG560000 BlueGreen Water Technologies OMB No.2040-0004
▪-g 2.5 Facility Location
• . Street,route number,or other specific identifier
Q 85 Mattamuskeet Rd.
c c County name County code(if known)
2 v Hyde
4 City or town State ZIP code
z 4T, Swanquarter NC 27885
SECTION 3. SIC AND NAICS CODES(40 CFR 122.21(f)(3))
3.1 SIC Code(s) Description(optional)
2879 Pesticides and Agricultural
v
0
U
v1
3.2 NAICS Code(s) Description(optional)
-0
325320 Pesticides and Other Agricultural Chemical
U _
N
SECTION 4.OPERATOR INFORMATION(40 CFR 122.21(f)(4))
4.1 Name of Operator
Jessica Frost
O 4.2 Is the name you listed in Item 4.1 also the owner?
o0 ❑ Yes ❑✓ No
• 4.3 Operator Status
0 Public—federal ❑Public—state El Other public(specify)
O ❑✓ Private ❑ Other(specify)
4.4 Phone Number of Operator
[phone]
4.5 Operator Address
Street or P.O.Box
E 14591 NW 21st Place
c City or town State ZIP code
o▪• o Newberry FL 32669
a Email address of operator
O [e-mail]
SECTION 5. INDIAN LAND(40 CFR 122.21(f)(5))
c 5.1 Is the facility located on Indian Land?
c ❑ Yes ✓❑ No
EPA Form 3510-1(revised 3-19) Page 2

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EPA Identification Number NPDES Permit Number Facility Name Form Approved 03/05/19
93647 NCG560000 BlueGreen Water Technologies OMB No.2040-0004
SECTION 6.EXISTING ENVIRONMENTAL PERMITS(40 CFR 122.21(f)(6))
6.1 Existing Environmental Permits(check all that apply and print or type the corresponding permit number for each)
ro
❑ NPDES(discharges to surface ❑ RCRA(hazardous wastes) ❑ UIC(underground injection of
water) fluids)
o co
• a ElPSD(air emissions) ❑ Nonattainment program(CAA) ❑ NESHAPs(CAA)
c _
0 Ocean dumping(MPRSA) ❑ Dredge or fill(CWA Section 404) 0 Other(specify)
SECTION 7. MAP(40 CFR 122.21(f)(7))
7.1 Have you attached a topographic map containing all required information to this application?(See instructions for
specific requirements.)
El Yes ❑ No ❑CAFO—Not Applicable(See requirements in Form 2B.)
SECTION 8. NATURE OF BUSINESS(40 CFR 122.21(f)(8))
8.1 Describe the nature of your business.
w
0
w
z
SECTION 9.COOLING WATER INTAKE STRUCTURES(40 CFR 122.21(f)(9))
9.1 Does your facility use cooling water?
m ElYes ElNo 4SKIP to Item 10.1.
t g9.2 Identify the source of cooling water.(Note that facilities that use a cooling water intake structure as described at
2 40 CFR 125,Subparts I and J may have additional application requirements at 40 CFR 122.21(r).Consult with your
•— NPDES permitting authority to determine what specific information needs to be submitted and when.)
Y
C NA
SECTION 10.VARIANCE REQUESTS(40 CFR 122.21(f)(10))
10.1 Do you intend to request or renew one or more of the variances authorized at 40 CFR 122.21(m)?(Check all that
apply.Consult withyour NPDES permittingauthorityto determine what information needs to be submitted and
PPly
when.)
c ❑ Fundamentally different factors(CWA ❑ Water quality related effluent limitations(CWA Section
d
Section 301(n)) 302(b)(2))
❑ Non-conventional pollutants(CWA ❑ Thermal discharges(CWA Section 316(a))
Section 301(c)and(g))
❑✓ Not applicable
EPA Form 3510-1(revised 3-19) Page 3
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EPA Identification Number NPDES Permit Number Facility Name Form Approved 03/05119
93647 NCG560000 BlueGreen Water Technologies OMB No.2040-0004
SECTION 11.CHECKLIST AND CERTIFICATION STATEMENT(40 CFR 122.22(a)and(d))
11.1 In Column 1 below,mark the sections of Form 1 that you have completed and are submitting with your application.
For each section,specify in Column 2 any attachments that you are enclosing to alert the permitting authority. Note
that not all applicants are required to provide attachments.
Column 1 Column 2
❑✓ Section 1:Activities Requiring an NPDES Permit ❑ wl attachments
❑✓ Section 2:Name,Mailing Address, and Location ❑ wl attachments
❑✓ Section 3:SIC Codes ❑ wl attachments
✓❑ S- ection 4:Operator Information ❑ w/attachments
✓❑ S- ection 5:Indian Land ❑ w/attachments
❑ Section 6:Existing Environmental Permits ❑ w/attachments
w/
0Section 7:Map
❑ map topographic ❑✓ w/additional attachments
El Section 8:Nature of Business ❑ wl attachments
❑ Section 9:Cooling Water Intake Structures ❑ w/attachments
U 0 Section 10:Variance Requests ❑ w/attachments
w ✓❑ Section 11:Checklist and Certification Statement ❑ wl attachments
11.2 Certification Statement
U
I certify under penalty of law that this document and all attachments were prepared under my direction or supervision
in accordance with a system designed to assure that qualified personnel properly gather and evaluate the
information submitted. Based on my inquiry of the person or persons who manage the system.or those persons
directly responsible for gathering the information,the information submitted is,to the best of my knowledge and
belief true. accurate, and complete.I am aware that there are significant penalties for submitting false information,
including the possibility of fine and imprisonment for knowing violations.
Name(print or type first and last name) Official title
Jessica Frost Scientific Director
Signature l � Gate signed
fi6dt 08/16/2023
EPA Form 3510-1(revised 3-19) Page 4

United States Office of Water EPA Form 3510-2E
Environmental Protection Agency Washington,D.C. Revised March 2019
Water Permits Division
,EPA Application Form 2E
Manufacturing, Commercial,
Mining, and Silvicultural
Facilities Which Discharge
Only Nonprocess Wastewater
NPDES Permitting Program
Note: Complete this form and Form 1 if your facility is a new or existing manufacturing, commercial,
mining,and silvicultural facility that discharges only nonprocess wastewater.

Paperwork Reduction Act Notice
The U.S. Environmental Protection Agency estimates the average burden to collect and complete
Form 2E to be 13.5 hours. This estimate includes time for reviewing instructions, searching existing data
sources, gathering and maintaining the needed data,and completing and reviewing the collection of
information. Send comments about the burden estimate or any other aspect of this collection of
information to the Chief, Information Policy Branch(PM-223), U.S. Environmental Protection Agency,
1200 Pennsylvania Avenue, NW, Washington, DC 20460,and to the Office of Information and
Regulatory Affairs,Office of Management and Budget, 725 17t Street, Washington, DC 20503,marked
"Attention: Desk Officer for EPA."

FORM 2E—INSTRUCTIONS
General Instructions
Who Must Complete Form 2E? Follow-up Requirements for New Dischargers
You must complete Form 2E if you answered"Yes"to Item 1.2.4 on Note that no later than 24 months after commencement of
Form 1—that is,if you are a new or existing facility(including discharge from the proposed facility,you must complete and
manufacturing,commercial.mining, and silvicultural facilities)that submit Section 4 of this form.At that time you must test and report
actual rather than estimated data for the pollutants or parameters
discharges only nonprocess wastewater. listed,unless waived by the NPDES permitting authority
Where to File Your Completed Form
Submit your completed application package(Forms 1 and 2E)to Line-by-Line Instructions
your National Pollutant Discharge Elimination System(NPDES) If you have multiple outfalls,you must submit a separate Form 2E
permitting authority.Consult Exhibit 1-1 of Form 1's"General for each(Sections 1.3,and 4 only).
Instructions"to identify your NPDES permitting authority. Section 1.Outfall Location
Public Availability of Submitted Information Item 1.1.Complete sections 1 through 6 for each outfall.Provide
The U.S.Environmental Protection Agency(EPA)will make the latitude and longitude to the nearest 15 seconds for the outfall.
information from NPDES permit application forms available to the Latitude and longitude coordinates may be obtained in a variety of
public for inspection and copying upon request.You may not claim ways,including use of hand held devices(e.g.,a GPS enabled
any information on Form 2E(or related attachments)as smartphone),internet mapping tools(e.g.,
confidential. https://mynasadata.larc.nasa.gov/latitudelongitude-finder/),
You may make a claim of confidentiality for any information that you geographic information systems(e.g..ArcView),or paper maps
submit to EPA that goes beyond the information required by Form from trusted sources(e.g.,U.S.Geological Survey or USGS).The
2E.Note that NPDES permitting authorities will deny claims for location of each outfall(i.e.,where the coordinates are collected)
treating any effluent data as confidential.If you do not assert a shall be the point where the discharge is released into a water of
claim of confidentiality at the time you submit your information to the United States.If you need further guidance in responding to
the NPDES permitting authority.EPA may make the information Item 1.1,refer to http://www.epa.govigeospatial/latitudelongitude-
available to the public without further notice to you.EPA will handle data-standard.
claims of confidentiality in accordance with the Agency's business Section 2.Discharge Date
confidentiality regulations at Part 2 of Title 40 of the Code of
Federal Regulations(CFR). Item 2.1.Indicate whether you are a new or an existing discharger.
Completion of Forms If you are an existing discharger,skip to Section 3 after completing
this item.
Print or type in the specified areas only. If you do not have enough
space on the form to answer a question,you may continue on Item 2.2.Indicate the date on which the facility will or is estimated
additional sheets.as necessary,using a format consistent with the to commence discharge.
form. Section 3.Waste Types
Provide your EPA Identification Number from the Facility Registry Item 3.1.Indicate the general type(s)of wastes being discharged or
Service, NPDES permit number, and facility name at the top of to be discharged,depending on whether you are an existing or new
each page of Form 2E and any attachments. If you do not know discharger.If you mark the response"Other Nonprocess
your EPA Identification Number,contact your NPDES permitting Wastewater."specify the nature of your discharge.
authority.See Exhibit 1-1 of Form 1's"General Instructions"for
contact information. Item 3.2.Indicate if the facility uses cooling water additives.If yes.
Do not leave any response areas blank unless the form directs you continue. If no,skip to Section 4.
to skip them.If the form directs you to respond to an item that does Item 3.3.List the cooling water additives being used(or to be used)
not apply to your facility or activity,enter"NA"for"not applicable"to and specify the composition of the additives,if such information is
demonstrate that you considered the item and determined a available to you.You can generally find composition information on
response was not necessary for your facility. product labels or from manufacturers'data sheets.
The NPDES permitting authority will consider your application Section 4.Effluent Characteristics
complete when it and any supplementary material are received and Items 4.1 to 4.8.These items require you to collect and report data
completed according to the authority's satisfaction.The NPDES for the parameters and pollutants listed in Section 4.The
permitting authority will judge the completeness of any application instructions are distinct for applicants with existing discharges
independently of the status of any other permit application or permit
for the same facility or activity. versus applicants that are new.
Definitions Important note: Read the"General Instructions for Reporting,
Sampling,and Analysis'on pages 2E-3 and 2E-4 before
The legal definitions of all key terms used in these instructions and completing Section 4.
Form 2E are in the"Glossary"at the end of the"General
Instructions"in Form 1.
2E-1

FORM 2E—INSTRUCTIONS CONTINUED
Item 4.1.Indicate whether you have completed monitoring for all limitations.If you wish to demonstrate your eligibility for a"net"
parameters in the table under Item 4.2 and attached it to the effluent limitation(i.e.,an effluent limitation adjusted to provide
application package.If you answer"No"because you have credit for the pollutant(s)present in your intake water)add a short
requested a waiver from your NPDES authority, skip to Section 5.If statement as to why you believe you are eligible.See also 40 CFR
"Yes,"continue to Item 4.2. 122.45(g).You will be contacted by the NPDES permitting authority
Item 4.2.Provide the sampling data requested in the table per the with further instructions.
-General Instructions for Reporting,Sampling.and Analysis"for Section 8.Checklist and Certification Statement
biochemical oxygen demand(BOD),total suspended solids(TSS), Item 8.1.Review the checklist provided on the application.In
oil and grease,ammonia(as N),flow,pH,and temperature(winter Column 1,mark the sections of Form 2E that you have completed
and summer). and are submitting with your application.For each section in
Item 4.3.Answer whether you believe fecal coliform to be present Column 2,indicate whether you are submitting attachments.
in your discharge or whether sanitary waste is discharged(or will be Item 8.2.The Clean Water Act(CWA)provides for severe penalties
discharged).If you answer"No."skip to Item 4.5.Otherwise, for submitting false information on this application form.CWA
continue to Item 4.4. Section 309(c)(2)provides that"Any person who knowingly makes
Item 4.4.Provide the sampling data requested in the table per the any false statement,representation,or certification in any
"General Instructions for Reporting,Sampling,and Analysis"for application, ...shall upon conviction,be punished by a fine of no
fecal coliform,Escherichia coli(E. colt),and enterococci. more than$10,000 or by imprisonment for not more than six
Item 4.5.Indicate whether chlorine is used(or will be used). If no, months,or both."
skip to Item 4.7.Otherwise.continue to Item 4.6. FEDERAL REGULATIONS AT 40 CFR 122.22 REQUIRE THIS
Item 4.6.Provide the sampling data requested in the table per the APPLICATION TO BE SIGNED AS FOLLOWS:
"General Instructions for Reporting,Sampling,and Analysis"for A. For a corporation,by a responsible corporate officer.For the
total residual chlorine. purpose of this section.a responsible corporate officer means:
Item 4.7.Answer whether non-contact cooling water is(or will be) (1)a president.secretary,treasurer,or vice-president of the
discharged from your facility.If no,skip to Section 5. If yes, corporation in charge of a principal business function,or any
continue to Item 4.8. other person who performs similar policy-or decision-making
Item 4.8.Provide the sampling data requested in the table per the functions for the corporation,or(2)the manager of one or
"General Instructions for Reporting, Sampling.and Analysis"for more manufacturing,production.or operating facilities,
chemical oxygen demand(COD), and total organic carbon(TOC). provided the manager is authorized to make management
decisions which govern the operation of the regulated facility
Section 5.Flow including having the explicit or implicit duty of making major
Item 5.1.Indicate whether any of the discharges that you described capital investment recommendations,and initiating and
in Sections 1 and 3(except for stormwater runoff,leaks,or spills) directing other comprehensive measures to assure long term
are intermittent or seasonal.If yes,continue to Item 5.2.If no, skip environmental compliance with environmental laws and
to Section 6. regulations;the manager can ensure that the necessary
Item 5.2.Describe the average frequency of flow and duration of systems are established or actions taken to gather complete
any intermittent or seasonal discharge(except for stormwater and accurate information for permit application requirements:
runoff, leaks,or spills)in gallons or million gallons per day(gpd or and where authority to sign documents has been assigned or
mgd),whichever is appropriate.The frequency of flow is the delegated to the manager in accordance with corporate
number of days or months per year there is an intermittent procedures.
discharge.Duration is the number of days or hours per discharge. B. For a partnership or sole proprietorship,by a general partner
For new dischargers.report your best estimate. or the proprietor,respectively.
Section 6.Treatment System C. For a municipality,state,federal,or other public facility,by
Item 6.1.Briefly describe any treatment system(s)used(or to be either a principal executive officer or ranking elected official.
used for new dischargers),indicating whether the treatment system For purposes of this section,a principal executive officer of a
is physical,chemical,biological,sludge and disposal,or other.Also federal agency includes:(1)The chief executive officer of the
give the particular type(s)of process(es)used(or to be used).For agency,or(2)a senior executive officer having responsibility
example,if a physical treatment system is used(or will be used), for the overall operations of a principal geographic unit of the
specify the processes applied(or to be applied),such as grit agency(e.g..Regional Administrators of EPA).
removal,ammonia stripping,dialysis,etc. END
Section 7.Other Information Submit your completed Form 1,Form 2E,and
Item 7.1.OPTIONAL ITEM. Report any additional information or all associated attachments
data(such as sampling results)that you believe the NPDES (and any other required NPDES application forms)
permitting authority should consider when establishing permit to your NPDES permitting authority.
2E-2

General Instructions for Reporting,Sampling,and Analysis
Important note:Read these instructions before completing Section If you have sampling and analysis questions.direct them to your
4 of Form 2E. NPDES permitting authority.The authority may request that you do
General Items additional testing,if appropriate,on a case-by-case basis under
CWA Section 308.
Complete the applicable tables for each outfall at your facility. Be
sure to note the EPA Identification Number,NPDES permit number, New Dischargers
facility name.and applicable outfall number at the top of each page You must provide maximum daily and average daily discharge
of any associated attachments. estimates for the parameters or pollutants listed in Section 4,unless
You may report some or all of the required data by attaching specifically indicated on the form. Note that if you have the results
separate sheets of paper instead of completing Section 4 for each of actual analyses for the listed parameters or pollutants,you are
of your outfalls so long as the sheets contain all of the required required to report those results rather than submit estimates.
information and are similar in format to Section 4. Report or estimate all parameter or pollutant levels as concentration
Reporting of Effluent Data and as total mass,except for flow,pH,and temperature. Indicate
Report pollutant levels for all pollutants in Section 4 as the source of all estimates in the appropriate column in the
concentration and total mass,with the exception of flow.pH.and Section 4 tables using the engineering study codes below. Note
temperature.Total mass is the total weight of pollutants that you are required to conduct follow-up testing and reporting no
discharged over a day. later than two years once your facility commences discharge.
Engineering Report Codes
Flow,temperature.pH,and fecal coliform organisms must be
g g P
Isis(°C),standard units,and most Actual data from pilot plants 1
reported as mgd,degrees Celsius
probable number per 100 milliliters(MPN/100 mL),respectively. Estimates from other engineering reports 2
Use the following abbreviations in the columns requiring"units"in Data from other similar plants 3
Section 4. Best professional estimates 4
Concentration Mass Others specify on the form
ppm=parts per million lbs=pounds Base your determination of whether a pollutant will be present in
mg/L=milligrams per liter ton=tons(English tons) your discharge on your knowledge of the proposed facility's use of
ppb=parts per billion mg=milligrams
maintenance chemicals and any analyses of your effluent or of any
pg/L=micrograms per liter g=grams similar effluent.You may also provide the estimates based on
available in-house or contractor engineering reports or any other
MPN=most probable number per kg=kilograms studies performed on the proposed facility.
100 milliliters T=tonnes(metric tons)
Pollutants Solely in Intake Water
Existing Dischargers
If you expect a pollutant to be present solely because of its
You must provide at least one analysis for each parameter or presence in your intake water,you must still provide an estimate or
pollutant,including the following: BOD,TSS,oil and grease, analytical result in Section 4;however,you should indicate in
ammonia(as N),fecal coliform including E. coli and enterococci(if Section 7 in Item 7.1 that you believe the pollutant or parameter to
believed present or if sanitary waste is or will be discharged).total be present only due to its presence in your source water.See the
residual chlorine(if chlorine is or will be used),COD,and TOC(if instructions under Item 7.1.
non-contact cooling water is or will be discharged),discharge flow, Testing Waivers
pH,and temperature(winter and summer).
You may report quantitative data that you have collected over the The NPDES permitting authority may waive the testing and
past 365 days if they are representative of your current operations. reporting requirements for flow or any of the pollutants listed in
The data reported must include maximum daily discharge,average Section 4 if you submit a written request for such a waiver before
daily discharge, and number of analyses.Most existing facilities or with your application.Contact your NPDES permitting authority
routinely monitor the pollutants and parameters listed in Section 4 for more information.
as part of their existing NPDES permit requirements. Sampling
You must collect and analyze samples in accordance with 40 CFR The collection of samples for the reported analyses should be
136.Grab samples must be used for analyses of pH,temperature, supervised by a person experienced in performing sampling of
total residual chlorine,oil and grease,fecal coliform(including E. industrial wastewater.You may contact your NPDES permitting
toll and enterococci(previouslyknown as fecal streptococcus) authorityfor detailedguidance on sampling techniques and for
), P ) P 9 si
and volatile organic compounds.Twenty-four-hour composite answers to specific questions.See Exhibit 1-1 of Form 1 for
samples must be used for all other pollutants,using at least four contact information.Any specific requirements in the applicable
grab samples unless otherwise specified at 40 CFR 136. For a analytical methods—for example,sample containers,sample
composite sample.only one analysis of the composite of aliquots is preservation,holding times,and the collection of duplicate
required. samples—must be followed,
2E-3

General Instructions for Reporting,Sampling,and Analysis Continued
The time when you sample should be representative of your normal • The method has the lowest ML of the analytical methods
operation,to the extent feasible,with all processes that contribute approved under 40 CFR 136 or required under 40 CFR
wastewater in normal operation.and with your treatment system chapter I,subchapter N or 0 for the measured pollutant or
operating properly with no system upsets.Collect samples from the pollutant parameter.
center of the flow channel,where turbulence is at a maximum, at a
specified inyourpresent NPDESpermit,or at anysite Consistent with 40 CFR 136,you may provide matrix-or sample-
site
adequate for the collection of a representative sample. specific MLs rather than the published levels.Further,where you
can demonstrate that.despite a good faith effort to use a method
Analysis that would otherwise meet the definition of"sufficiently sensitive,"
Except as specified below,all required quantitative data shall be the analytical results are not consistent with the quality assurance
collected in accordance with sufficiently sensitive analytical (QA)/quality control(QC)specifications for that method,then the
methods approved under 40 CFR 136 or required under 40 CFR NPDES permitting authority may determine that the method is not
chapter I,subchapter N or 0.A method is"sufficiently sensitive" performing adequately and the NPDES permitting authority should
when: select a different method from the remaining EPA-approved
methods that is sufficiently sensitive consistent with 40 CFR
• The method minimum level(ML)is at or below the level of the 122.21(e)(3)(i).Where no other EPA-approved methods exist,you
applicable water quality criterion for the measured pollutant or must select a method consistent with 40 CFR 122.21(e)(3)(ii).
pollutant parameter, When there is no analytical method that has been approved under
• The method ML is above the water quality criterion,but the 40 CFR 136;required under 40 CFR chapter I,subchapter N or 0,
amount of the pollutant or pollutant parameter in the facility's and is not otherwise required by the NPDES permitting authority,
discharge is high enough that the method detects and you may use any suitable method but shall provide a description of
quantifies the level of the pollutant or pollutant parameter in the method.When selecting a suitable method,other factors such
the discharge. as a method's precision,accuracy,or resolution,may be
considered when assessing the performance of the method.
2E-4

f

EPA Identification Number NPDES Permit Number Facility Name Form Approved 03/05/19
93647 NCG560000 BlueGreen Water Technologies OMB No 2040-0004
U.S.Environmental Protection Agency
FORM Application for NPDES Permit to Discharge Wastewater
2E NPDES EPA MANUFACTURING,COMMERCIAL,MINING,AND SILVICULTURAL FACILITIES WHICH
DISCHARGE ONLY NONPROCESS WASTEWATER
SECTION 1,OUTFALL LOCATION(40 CFR 122,21(h)(1))
1.1 Provide information on each of the facikty's outfalls in the table below.
Outfall Receiving Water Name Latitude Longitude
Number
0
o
O
•
SECTION 2. DISCHARGE DATE(40 CFR 122.21(h)(2))
2.1 Are you a new or existing discharger?(Check only one response.)
.20
✓❑ New discharger ❑ Existing discharger 4 SKIP to Section 3.
2.2 Specify your anticipated discharge date: 10/23/2023
SECTION 3 WASTE TYPES(40 CFR 122.21(h)(3))
3.1 What types of wastes are currently being discharged if you are an existing discharger or will be discharged if you are a
new discharger?(Check all that apply.)
❑ Sanitary wastes El Other nonprocess wastewater(describe/explain
❑ Restaurant or cafeteria waste directly below)
m El Non-contactcooling water Sodium percarbonate
n.
3.2 Does the facility use cooling water additives?
❑ Yes ✓❑ No 4 SKIP to Section 4.
3.3 List the cooling water additives used and describe their com?osition.
Cooling Water Additives Composition of Additives
(list) (if available to you)
SECTION 4. EFFLUENT CHARACTERISTICS(40 CFR 122.21(h)(4))
4.1 Have you completed monitoring for all parameters in the table below at each of your outfalls and attached the results to
this application package?
El Yes No:a waiver has been requested from my NPDES permitting authority
(attach waiver request and additional information)4 SKIP to Section 5.
4.2 Provide data as requested in the table below.' (See instructions for specifics)
Number of Maximum Daily Average Daily Source
Parameter or Pollutant Analyses Discharge Discharge (use codes
(if actual data (specify units) (specify units) per
instructions
reported) Mass Conc. Mass Cont. )
m
Biochemical oxygen demand(BOD=.)
c
Total suspended solids(TSS)
Oil and grease
Ammonia(as N)
Discharge flow
pH(report as range)
Temperature(winter)
Temperature(summer)
'Sampling shalt be conducted according to sufficiently sensitive test procedures(i.e.,methods)approved under 40 CFR 136 for the analysis of pollutants or pollutant
parameters or required under 40 CFR chapter I,subchapter N or 0 See instructions and 40 CFR 122.21(eX3).
EPA Form 3510-2E(revised 3-19) Page 1

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EPA Identification Number NPDES Permit Number Facility Name Form Approved 03/05/19
93647 NCG560000 BlueGreen Water Technologies OMB No 2040-0004
4.3 Is fecal coliform believed present,or is sanitary waste discharged(or will it be discharged)?
❑ Yes ❑ No 4 SKIP to Item 4.5.
4.4 Provide data as requested in the table below.' (See instructions for specifics.)
Number of Maximum Daily Average Daily Source
Parameter or Pollutant Analyses Discharge Discharge (Use codes
(if actual data (specify units) (specify units) per
reported) Mass Conc. Mass Conc. Instructions.)
Fecal coliform
0.)
E.coil
= Enterococci
0 4.5 Is chlorine used(or will it be used)?
❑ Yes ❑ No 4 SKIP to Item 4.7.
4.6 Provide data as requested in the table below.' (See instructions for specifics.)
Number of Maximum Daily Average Daily Source
Analyses Discharge Discharge (use codes
✓ Parameter or Pollutant (if actual data (specif; units) (specify units) per
reported) Mass Conc. Mass Conc. instructions)
Total Residual Chlorine
w 4.7 Is non-contact cooling water discharged(or will it be discharged)?
❑ Yes ❑ No 4 SKIP to Section 5.
4.8 Provide data as requested in the table below.'(See instructions for specifics.)
Number of Maximum Daily Average Daily Source
Parameter or Pollutant Analyses Discharge Discharge (use codes
(if actual data (specify units) (specify units) per
reported) Mass Conc. Mass Conc. instructions)
Chemical oxygen demand(COD)
Total organic carbon(TOC)
SECTION 5. FLOW(40 CFR 122.21(h)(5))
5.1 Except for stormwater water runoff,leaks,or spills,are any of the discharges you described in Sections 1 and 3 of this
application intermittent or seasonal?
❑ Yes 4 Complete this section. ❑ No 4 SKIP to Section 6.
0 5.2 Briefly describe the frequency and duration of flow.
SECTION 6.TREATMENT SYSTEM(40 CFR 122.21(h)(6))
6.1 Briefly describe any treatment system(s)used(or to be used).
rc
'Sampling shall be conducted according to sufficiently sensitive test procedures(i.e.,methods)approved under 40 CFR 136 for the analysis of pollutants or pollutant
parameters or required under 40 CFR chapter I,subchapter N or 0.See instructions and 40 CFR 122.21(eX3).
EPA Form 3510-2E(revised 3-19) Page 2

l

EPA Identification Number NPDES Permit Number Facility Name Form Approved 03/05/19
93647 NCG560000 BlueGreen Water Technologies OMB No 2040-0004
SECTION 7.OTHER INFORMATION(40 CFR 122.21(h)(7))
7.1 Use the space below to expand upon any of the above items. Use this space to provide any information you believe the
reviewer should consider in establishing permit limitations.Attach additional sheets as needed.
0
fa
0
6
SECTION 8. CHECKLIST AND CERTIFICATION STATEMENT(40 CFR 122.22(a)and(d))
8.1 In Column 1 below,mark the sections of Form 2E that you have completed and are submitting with your application.
For each section,specify in Column 2 any attachments that you are enclosing to alert the permitting authority.Note that
not all applicants are required to provide attachments.
Column 1 Column 2
❑� Section 1:Outfall Location ❑ w/attachments(e.g.,responses for additional outfalls)
❑✓ Section 2:Discharge Date CI wf attachments
O Section 3:Waste Types ❑ w/attachments
Section 4:Effluent Characteristics ❑ wi attachments
❑✓ Section 5:Flow 0 w/attachments
in
o ❑✓ Section 6:Treatment System ❑ wl attachments
© Section 7:Other Information ❑ w/attachments
r, © Section 8:Checklist and Certification Statement 0 w/attachments
8.2 Certification Statement
I certify under penalty of law that this document and all attachments were prepared under my direction or supervision in
CD
accordance with a system designed to assure that qualified personnel properly gather and evaluate the information
submitted. Based on my inquiry of the person or persons who manage the system,or those persons directly
responsible for gathering the information,the information submitted is,to the best of my knowledge and belief.true.
accurate,and complete. I am aware that there are significant penalties for submitting false information,including the
possibility of fine and imprisonment for knowing violations.
Name(print or type first and last name) Official title
Jessica Frost Scientific Director
Signature / - -� Date signed
08/16/2023
EPA Form 3510-2E(revised 3-19) Page 3

•
- i 1 }_" of '
# ,--
J y

EPA ldentrfication Number NPDF.S Number Facility Name —Outtali Number
93647 NCG560000 6 lueGreen Water Technclo-,es 1
Method Number Estimated Concentration Of
Pollutant(Required) CAS number (if Applicable) Reason Pollutant Believed Present in Discharge Known)
' Sodium parcarbonate 15630-89)4 Acttve ingredient in algaecide at 83.3% 0.18.36.26 ppm
•
•
•
•
•
I _
I
•
,.//7) •
0

Pesticide Discharge Management Plan
� g
(PDMP)
RECEIVED
BlueGreen US Water Technologies, Inc. APR 12 2024
Mailing Address:301 South Hills Village Ste LL200#452
Pittsburgh,PA 15241 NCDEQ/DWR/NPDES
Decision-maker(s):
Permit and Treatment Decision Makers(Manager, Supervisor, Owner, etc):
Dr. Jessica Frost—US Science Director
BlueGreen US Water Technologies, Inc.
jessica.frost a(�.bluegreenwatertech.com
Cell: 941.224.2218
Person(s) Responsible for PDMP Development and Implementation:
Dr. Jessica Frost—US Science Director
BlueGreen US Water Technologies, Inc.
jessica.frost bluegreenwatertech.com
Cell: 941.224.2218

PDMP Preparation/Revision Date:Aug 29,2023 POMP Implementation Date:June/July 2024
1. PDMP Team
a. Persons responsible for managing pests in the pest management area?
Dr.Jessica Frost
*Pests is defined as cyanobacteria(aka bluegreen algae)
*Management area defined as Lake Mattamuskeet, North Carolina
b. Persons responsible for developing and revising the PDMP?
Dr.Jessica Frost
c. Persons responsible for developing, revising, and implementing corrective actions and other
effluent limitation requirements?
Dr.Jessica Frost
d. Persons/Teams/Companies responsible for pesticide applications?
Dr.Jessica Frost(BlueGreen)
BlueGreen is hiring applicators as subcontractors. Once the permit application is approved, a
subcontractor will be selected. List of potential subcontractors:
Modica& Associates
SOLitude
WSB
2. Pest Management Area Description
a. Pest Problem Description.
Lake Mattamuskeet is the largest natural freshwater lake in North Carolina. It lies next to
the Pamlico Sound in eastern North Carolina, The 40,000 acre lake makes up the majority
of the Mattamuskeet National Wildlife Refuge, which is managed by USFWS. The lake is
nearly centrally located along the Atlantic Flyway, a route migratory birds use to move south
for the winter. The refuge is extremely valuable for wintering waterfowl to stop, rest and eat
along their journey. According to USFWS, the refuge attracts more than 200,000 ducks,
geese and swans from November through February. About 58,000 people visit the refuge
annually to observe the birds, or to hunt and fish in the summer. For rural Hyde County, this
brings important economic stimulation. Since the 1980's, water quality declines have been
monitored in the lake. Results consistently show pH and chlorophyll a levels above normal
limits, indicating algae blooms in the water. These blooms are caused by excess nutrients
in the water. such as nitrogen and phosphorus. Toxic algal blooms (aka HABs) have also
been monitored. These blooms have some of the highest concentrations of algal toxins in
the country, bordering on federal limits for recreational contact. In 2016, the lake was listed

on the state 303(d) list for impaired waters due to elevated levels of pH and chlorophyll-a.
The water quality within the lake has drastically declined due to significant increases in
nutrients and suspended sediments that have been contributed from over a century of
landscape alterations and hydrologic modifications. In addition, monitoring by the U.S. Fish
and Wildlife Service (USFWS) has indicated that most of the submerged aquatic vegetation
(SAV), an important habitat for fish and food source for waterfowl was lost in the lake by
2017. The algae blooms are blocking sunlight from making it to the lake bottom. When this
happens for long periods of time, grass begins to disappear. The source of these HABs is
cyanobacteria, which is known to produce toxins of varying levels. Monitoring results also
indicates algae blooms containing a cyanotoxin, cylindrospermopsin, at concentrations that
border the federal limits for recreational contact has become a more frequent occurrence.
Note: Information herein is referenced in the 2018 Lake Mattamuskeet Watershed
Restoration Plan.
b. Action Thresholds.
In collaboration with University of North Carolina (UNC) Collaboratory, subject matter
experts from UNC Institute for Marine Sciences, and the US Fish and Wild Service,
BlueGreen is engaged in monitoring to establish treatment dosages in alignment with best
practices and the product label. Our research project includes monitoring using remote
satellite-based analysis, in-situ field measurements and direct water sampling with
laboratory analysis provided by UNC IMS.
Product Label Excerpt:
APPLICATION RATES
The best method by which to apply Lake Guard` Oxy granules to water is by broadcasting Idusting)it
over a well-defined contamination zone,at early bloom stages,when harmful algat'cyanobacteria
numbers are at 5.000 to 20.000 cells mi. and below 10 ug chk rophylt-alder)
Determination of surface area to be treated
Determine the size of the infested area as follows:(1)in small infested reservoirs.under than 250 acres.
obtain surface area by measuring of regular shaped ponds or mapping of irregular ponds or by reference
to previously recorded engineering data or maps.(2)In water bodies larger than 250 acres(or smaller
ponds with a defined contaminated zone)outline the infested area by a combination of the following
instruments.microscopical count,pigment extraction,toxin evaluation,probes that detect specific
pigments that are known to serve as a correlated proxy for algae'cyanobactena biomass.satellite
imaging.etc.NOTE:evaluation of the state of the infestation should be done by professional personnel
Determination of the application rate
For control of harmful algae;cyanobacteria infestation it is essential to begin Lake Guard'Oxy treatment
when harmful algaetcyanobactena cell numbers are in the range of 5.000-20,000 cell!mL(or below 10 pg
chlorophyll-a+liter) Apply 0.5-5 lbs+acre Lake Guard' Oxy at these algal cyanobacterial cell-densities
Always start with the lower rate.At higher infestation rates,when cyanobactenal cell density is between
20.000-100,000 cellsimL for between 10.50 pg chlorophyll-ander)use 5-301bs;acre Lake Guard"Oxy.
If treatment is delayed until algalicyanobactenal cell numbers exceed 100.000 cell.'mL(or equivalently.
above 50 pg chlorophyll-arlitor).an increase in the quantities of the Lake Guards'Oxy will be required,as
well as in treatment frequency.Therefore,in heavy blooms.when cyanobacterial scum or aggregates are
visible to the naked eye (more than 100.000 cellsJmL of algaeicyanobacteria or over 50 pg chtorophyli-
aMliter) treat with doses between 30.98 lbs.racre If doses exceed 98 lbs'acre.treat no more than one-
half of the water area in a single application Maximum single application rate allowed should not exceed
294 lbs'acre of the Lake Guard'Oxy.NOTE.when cyanobacterial aggregated could be seen with the
naked eye,the cyanobacterial cell density in the water is estimated to exceed 100.000 cells per ml,
When a single application dose is below 30 tbs.acre.minimum retreatment interval is 12 hours When a
single application dose is between 30-98 lbs'acre,minimum retreatment interval is 24 hours When a
single application dose exceeds 98 Ibs.'acre.the minimum retreatment interval is 48 hours.

c. Provide Location and Treatment Area Maps or refer to NOI maps.
Treatment area is defined as the water surface of Lake Mattamuskeet:
:41tlNP'CA'Y t
4 r, �..
r.M+ FFt004
k �
4, F
F s
TT) P,M'+
.0.a,d M1.r,.p»
4 n...h.
c • + »�rr.�.4
Figure 1°Map of Hyde County,N.0 General watershed area outlined
RESEARCH PROJECT PLAN SITE DETAILS:
Area Size Perimeter Size Turbidity Curtain Total Size
Site Location (Acres) (miles) (mites)
WEST BASIN
West Treatment A 131.0 2.8 0.1
West Control A 145.0 2.5 0.5
West Treatment B 40.6 1.1 0.3
West Control B 8.8 0.6 0.1
West Treatment Total Acres 171.6
West Site Locations Total
Acres 325.4
EAST BASIN
EastTreatmentA 212.0 2.5 0.6
East Control A 95.9 1.7 0.6
East Treatment B 6.5 0.4 0.1
East Control B 80.0 1.5 0.5
East Treatment Total Acres 218.5
East Site Locations Total
Acres 394.4

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d. Water Quality Standards.
Since this is a research project in collaboration between BlueGreen, UNC Collaboratory,
UNC IMS, and USFWS, the following is a comprehensive list of parameters that will be
collected and analyzed:
Chlorophyll a
Phycocyanin
Accessory pigments by HPLC
Phytoplankton species microscopy
Chlorophyll a fluorescence
Phycocyanin fluorescence
Nutrients (N and P)
Cylindrospermopsin toxin
Microcystin toxin
BMAA (emergent toxin of concern for eastern North Carolina)
Dissolved oxygen
pH
Zooplankton biomass/community structure
Turbidity
Colored dissolved organic matter
PAR attenuation
Salinity
Temperature
Remote sensing

I

3. Control Measure Description
BlueGreen includes our EPA registered product label and SDS as an attachment to this
document. The active ingredient in Lake Guard Oxy®is sodium percarbonate, which forms
hydrogen peroxide when mixed with water. The hydrogen peroxide is activated only on the
upper surface of the water and releases H2O and 02 molecules as byproducts. Control
measures are typically based on adherence to the EPA product label and optimizing
outcomes based on minimum application rates. not to exceed the EPA product label. In
collaboration and agreement amongst BlueGreen. UNC Collaboratory, UNC IMS, and
USFWS, this research project demonstrates due diligence beyond adherence to the EPA
product label by taking extra precautions that are specifically designed for Lake
Mattamuskeet. primarily driven by an abundance of caution. More specifically. BlueGreen
voluntarily performed a variety of EPA recognized and standardized toxicology tests that
included Ceriodaphnia dubia. Apis mellifera, and northern bobwhite quail (in lieu of mallard
ducks that were initially tested, however, because of a strong regurgitation response, and
LD50 was not possible to determine for mallard ducks). BlueGreen includes all toxicology
final reports as attachments to this document. As a result of the toxicology tests, the project
team created thresholds for dose application and contingencies for stopping treatment.
These thresholds and contingencies are outlined below as a part of the mitigation protocol
for Lake Mattamuskeet, as agreed by upon BlueGreen, UNC Collaboratory, UNC IMS. and
USFWS.
• LAKE GUARD OXY®TREATMENT CASE SCENARIOS
TREATMENT
AREAS
SCENARIO BASED ESTIMATED MONITORING
ON CYANOBACTERIA MAXIMUM MONITORING FREQUENCY
CELL DENSITY IF THEN DOSE USE METRICS (TIN HOURS)
1. CYANOBACTERIA LAKE GUARD WEST BASIN: CELL COUNTS @ TO,T12,T24,T36,T48,
CELL DENSITY OXY DOSE @ 15,600 LBS T72,T96,T120,T144 OR
>100,000 CELLS/ML 50LBS/ACRE EAST BASIN: UNTIL TARGET
(18.4 PPM) 14,600 LBS CYANOBACTERIA CELL
DENSITY 20,000
CELLS/ML IS MET,
WHICHEVER COMES
FIRST
2. CYANOBACTERIA LAKE GUARD WEST BASIN: CELL COUNTS @ TO,T12,T24,T36,T48,
CELL DENSITY OXY DOSE @ 9,360 LBS T72 T96 T120 T144 OR
>20,001-100,000 30LBS/ACRE EAST BASIN: UNTIL TARGET
CELLS/ML (11.04 PPM) 8,760 LBS CYANOBACTERIA CELL
DENSITY 20,000
CELLS/ML IS MET,
WHICHEVER COMES
FIRST
3. CYANOBACTERIA LAKE GUARD WEST BASIN: CELL COUNTS @ TO,T12,T24,T36,T48,
CELL DENSITY OXY DOSE @ 1,560 LBS T72,T96,T120,T144;A
>5,000-20,000 5LBSIACRE EAST BASIN: MAINTENANCE
CELLS/ML (1.84 PPM) 1.460 LBS STRATEGY TO ENSURE
CYANOBACTERIA CELL
DENSITY DOES NOT
EXCEED 20,001
CELLS/ML

FOR LAKE MATTAMUSKEET:
A TREATMENT EVENT IS DEFINED AS 2 WEEKS IN DURATION
PERIOD OF PERFORMANCE IS DEFINED AS APRIL 1-OCTOBER 31, 2024
THE MAXIMUM#OF TREATMENTS BASED ON THE ABOVE IS 2 PER MONTH (OR A TOTAL OF 14)
*Note-Lake Guard Oxy®EPA product label stipulates the following as guidelines for retreatment instructions:
When a single application dose is below 30 bs/acre, minimum retreatment interval is 12 hours.
When a single application dose is between 31-_98 Ibs/acre, minimum retreatment interval is 24 hours.
When a single application dose exceeds x , minimum retreatment interval is M.
TREATMENT
AREAS
SCENARIO ESTIMATED
BASED ON MAXIMUM MONITORING MONITORING FREQUENCY
CYANOTOXINS IF THEN DOSE USE METRICS (T IN HOURS)
1. CYANOTOXINS LAKE GUARD WEST BASIN: TOXINS @ TO,T24,T48,T72,T96,T120,
>8 PPB OXY DOSE @ 15,600 LBS T144 OR UNTIL TARGET
50LBS/ACRE EAST BASIN: CYANOTOXINS 8 PPB IS MET,
(18.4 PPM) 14,600 LBS WHICHEVER COMES FIRST
• STOP TREATMENT CONTINGENCIES
CONTIGENCY
PLAN BASED ON MONITORING
DO IF THEN METRICS MONITORING FREQUENCY
1. DISSOLVED OXYGEN STOP DO YSI-CONTINUOUS DATA LOGGING
<4 MG/L FOR GREATER TREATMENT
THAN 8 HOURS
2 MG/LAT ANY TIME I
Lake Mattamuskeet estimated annual average DO: 106%or 9.56 mg/L.
CONTIGENCY
PLAN BASED ON MONITORING
PH IF THEN METRICS MONITORING FREQUENCY
2. PH>10.5 STOP PH YSI-CONTINUOUS DATA LOGGING
PH<6 TREATMENT
Lake Mattamuskeet 2023 pH ranged 7.5-9.
4. Schedules and Procedures
January 2023-present: Continuous baseline data collection and monitoring.
April 1-October 31,2024: Period of performance for mitigation activities.
July 2024:Target month for first mitigation treatment,followed by continuous monitoring and data
collection to guide subsequent actions,which may include re-treatment,or dose recommendation.
Please refer to the above treatment case scenarios for decision making.
a. Document Control Measures
1) Application rate and frequency. Include procedures for determining the lowest effective
amount of pesticide product per application and the optimum frequency of applications
necessary to control the pest while reducing the potential for pest resistance.
a. The EPA product label establishes baseline dosage amounts and frequencies.
Optimizing outcomes to establish lowest effective dosage/application is influenced

by our team of scientists and USFWS and in consideration of best practices,
including:
i. Time of day
ii. Wind and weather patterns
iii. Historical analysis and trend modeling
iv. Cell counts
v. Remote satellite-based monitoring
vi. In-situ probe measurements
b. The mode of action of the product is a slow, time-release of sodium percarbonate
that converts immediately to hydrogen peroxide when mixed with water and further
disassociates to molecules of water and oxygen. Hydrogen peroxide is a Reactive
Oxygen Species (ROS) that causes oxidative stress. to which cyanobacteria are
known to be the most susceptible and is accredited for triggering a Programmed
Cell Death (PCD)cycle within the cyanobacteria colony.
c. Frequency of application adheres to the EPA label and is influenced by the
cyanobacteria's movement within the vertical water column and overall
concentration of cell density.
2) Spill prevention. Procedures and schedule of maintenance for preventing spills and leaks
of pesticides associated with the application of pesticides.
a. Spill prevention procedures are established by adhering to the EPA product label,
the Safety Data Sheet(SDS) and ensuring the use of licensed applicators. (Safety
Data Sheet included as attachment).
3) Pesticide application equipment. Schedules and procedures for maintaining application
equipment in proper operating condition to prevent spills and overapplication, including
calibrating, cleaning, and repairing equipment.
a. BlueGreen will be contracting with a licensed subcontractor.
b. Licensed subcontractor(s) are responsible for the maintenance of internal fleet
records, insurances and certifications, and timely communications with project
manager.
4) Pest surveillance. Procedures and methods for conducting pre-application pest
surveillance.
a. BlueGreen has and is conducting field-based observations, remote monitoring
based on satellite imagery analysis, in-situ measurements, and water sampling.
Additionally, BlueGreen has analyzed historical datasets available from 3rd party
sources, such as USGS, Mattamuskeet National Wildlife Refuge. and the Lake
Mattamuskeet Technical Working Group, across multiple years.
b. UNC IMS has and is conducting monthly field-based operations for water quality.
5) Assessing environmental conditions. Procedures and methods for assessing
environmental conditions before and after treatments in the treatment area.
a. Both BlueGreen and UNC IMS continue to conduct field-based observations,
remote monitoring based on satellite imagery analysis, in-situ measurements, and
water sampling. Additionally. BlueGreen has analyzed historical datasets available
from 3rd party sources, such as USGS, Mattamuskeet National Wildlife Refuge, and
the Lake Mattamuskeet Technical Working Group, across multiple years.
b. Environmental conditions will include additional attributes. such as temperature,
dissolved oxygen. pH and similar water quality parameters (please refer to Section
2 d. for a comprehensive list of parameters).

b. Other Actions to Minimize Discharges.
1) Spill response procedures.
a. Procedures for expeditiously stopping, containing, and cleaning up leaks, spills, and
other releases. Employees who may cause, detect, or respond to a spill, leak, or
overapplication must be trained in these procedures and have necessary spill response
equipment available.
BlueGreen's spill response procedures are detailed within our Safety Data Sheet,which is
included as an attachment,with excerpt below.
6.ACCIDENTAL RELEASE MEASURES
i sonic Ensure adequate ventilation.Avoid contact with eyes or clothing-Evacuate personnel to safe areas.Keep people away
from and upwind of spdVleak_ELIMINATE all ignition sources(no smoking.flares,sparks.or flames in immediate area).
Do not touch damaged containers or spilled material unless wearing appropriate protective clothing. See section 8 for
more information. Stop leak if you can do it without risk. Use personal protective equipment as required. Keep
combustibles(wood,paper,oil,etc.)away from spilled material DO NOT GET WATER INSIDE CONTAINERS.
Ventilate the area Refer to protective measures listed in Sections 7 and 8.
Stop leak if you can do it without risk.Cover with DRY earth,DRY sand or other non-combustible material followed with
plastic sheet to minimize spreading or contact with rain
With clean shovel place material into dean,dry container and cover loosely:move containers from spill area Flush area
with flooding quantities of water Prevent product from entering drains.Cover powder spill with plastic sheet or tarp to
minimize spreading and keep powder dry
b. Procedures for notification of appropriate facility personnel,emergency response
agencies, and regulatory agencies.
BlueGreen's spill response procedures are detailed within our Safety Data Sheet, which
is included as an attachment. Any applicators shall be licensed accordingly and
onboarded to the SDS through routine best practices.
Additional points of contract shall include first responders (if needed), BlueGreen project
manager, USFWS who manages Lake Mattamuskeet, North Carolina Department of
Environmental Quality, Hyde County Health Department,and any other points of contact
that agencies deem necessary.
2) Adverse incident response procedures.
a. Procedures for responding to any incident resulting from pesticide, biological agents, or
chemical applications.
6.ACCIDENTAL RELEASE MEASURES
suers Ensure adequate ventilation.Avoid contact with eyes or clothing.Evacuate personnel to safe areas.Keep people away
from and upwind of spill/leak.ELIMINATE all ignition sources(no smoking.flares,sparks.or frames in immediate area>
Do not touch damaged containers or spilled material unless wearing appropriate protective clothing. See section 8 for
more information. Stop leak if you can do it without risk Use personal protective equipment as required. Keep
combustibles(wood,paper,oil,etc)away from spilled material DO NOT GET WATER INSIDE CONTAINERS.
Ventilate the area.Refer to protective measures listed in Sections 7 and 8.
Stop leak if you can do it without risk.Cover with DRY earth,DRY sand or other ran-combustible material followed with
plastic sheet to minimize spreading or contact with rain.
With dean shovel place material into dean.dry container and cover loosely,move containers from spill area.Flush area
with flooding quantities of water. Prevent product from entering drains.Cover powder spill with plastic sheet or tarp to
minimize spreading and keep powder dry.
b. Procedures for notification of the incident, both internal to your agency/organization and
external. Contact information for state/federal permitting agency, nearest emergency
medical facility, and nearest hazardous chemical responder must be in locations that
are readily accessible and available.
BlueGreen's spill response procedures are detailed within our Safety Data Sheet, which
is included as an attachment. Any applicators shall be licensed accordingly and
onboarded to the SDS through routine best practices,

1. PRODUCT 8 COMPANY IDENTIFICATION
'' Product Name BLUEGREEN ' LAKE GUARDTm OXY
I 2 Chemical Name Sodium Percarbonate Mixture
13 synonyms NA ------1 4 Tiede Name. BlueGreen Lake Guard'M Oxy _---
1 5 Produu u.e Algaeade/BiOcide
1.8 Distributor•Name: BlueGreen Water Technologies Ltd.
i r nrsnCular s Address 3/15 Kachal Street,Tzur Hadassah,9987500,Israel
1 8 Emergency Wane CHEMTEL+1 [phone]
t s Business Phone,Fes I Email Tel:+$72(2)830-1108 I Fax:[phone]/Email:infogb_sitechs corn
BlueGreen Points of Contact:
- Dr. Jessica Frost (Project Manager); c: 941.224.2218;
e: jessica.frost a(�bluegreenwatertech.com
- Mike Pelz (Project Contract Manager); c: 828.384.5146;
e: mike.pelzQbluegreenwatertech.com
Applicator Points of Contact:
TBD once permit approval is attained
USFWS Mattamuskeet National Wildlife Refuge Point of Contact:
- Kendall Smith (Refuge Manager); c: 252.473.0241; e: kendall smith(a)fws.gov
North Carolina Department of Environmental Quality Point of Contact
- Julie Grzyb, Deputy Director; p: Office: [phone] 1 Cell: [phone];
e: julie.grzvb anncdenr.gov
Hyde County Department of Health
- 1151 Main St., Swanquarter, North Carolina, 27885; p: 252.926.4399
Emergency Hospitals
9 Y P
- Engelhard Medical Center(open 0800-1700)
33270 US-264, Engelhard, North Carolina, 27824; p: 252.925.7000
- Washington Regional Medical Center(open 24 hours)
958 US-64, Plymouth, North Carolina, 27962; p: 252.793.4135
3 Pesticide monitoring schedules and procedures.
a The process for determining the location of any monitoring.
Lake Mattamuskeet will be monitored by a combination of in situ measurements. remote
satellite-based analytics, and monthly water quality sampling prior to mitigating
cyanobacteria. High frequency, approximately daily,water quality sampling monitoring will
be conducted immediately following treatment by resumption of twice monthly sampling for
the summer after treatment and monthly monitoring for fall and winter.
b. A schedule for monitoring.
Ongoing continuous,with the finest resolution of data collection occurringevery30 seconds via
9 9
in situ measurements.
c. The person (or title) responsible for conducting monitoring.
Dr. Jessica Frost—BlueGreen US Scientific Director
d. Procedures for documenting any observed impacts to non-target organisms resulting from
pesticide application/discharge.
Formal Report will be issued by Dr. Jessica Frost

5. Signature Requirements
Operators must sign, date,and certify the PDMP.
Certification Statement:"I certify under penalty of law that this document and all
attachments were prepared under my direction or supervision in accordance with a
system designed to assure that qualified personnel properly gathered and evaluated the
information contained therein. Based on my inquiry of the person or persons who
manage the system,or those persons directly responsible for gathering the information,
the information contained is,to the best of my knowledge and belief,true, accurate,and
complete. I am aware that there are significant penalties for submitting false information,
including the possibility of fine and imprisonment for knowing violations."
Authorized Person Signature:
Date:_Aug. 29,2023

FWS Form 3-1383-R(Rev.03/2020) OMB Control No. 1018-0102
U.S. Department of the Interior Expires 01/31/2024
r, .y RESEARCH AND MONITORING ,,,,�;�,,,,,
1r--T SPECIAL USE PERMIT
\ 1111111 / rivi
,.,,_:- N;,.►•• National Wildlife Refuge System
Refuge Name: Lake Mattamuskeet For Official Use Only
Address: 85 Mattamuskeet Road, Swan Quarter, NC 27885 Approved Permit'-'
Attn:(Refuge Official) Kendall Smith Station#:
E-Mail: [e-mail] 'ermit Term:from
Phone#: [phone]
Note:We do not require all information for each Research project. See instructions at the end of the notice and contact the refuge
to determine applicability of a particular item. Attach additional sheets if the text spaces provided are inadequate.
la) Identify the type of Permit you are applying for: New() Renewal O Modification O Other()
lb)Have you applied,or do you intend to apply,to any other refuges for this same activity? ()Yes O No
1c)If yes,which refuges?
Applicant Information
2)Principal investigator: Nathan Hall 3) Is curriculum vitae or resume attached? Yes O No()
4a)Affiliation/Sponsoring Organization: UNC Chapel Hill Institute of Marine Sciences, 3431 Arendell St.
4b)Relationship to affiliation/sponsoring organization(professor,staff, student,etc.): research assistant professor
5) Physical Address: UNC Chapel Hill Institute of Marine Sciences, 3431 Arendell St.
City/State/Zip:
Morehead City, NC 28557
6)Mailing Address:(if different than above)
City/State/Zip:
7) Phone#: [phone] 8) Fax#: 9)E-mail: [e-mail]
10)List known assistants/subcontractors/subpermittees: (Only required if the assistants/subcontractors/subpermittees will be operating on the refuge without
the permittee being present.)
Name Address Phone# —
Jeremy Braddy UNC Institute of Marine Sciences 252 726 6841
Chesney Thomspon UNC Institute of Marine Sciences 252 726 6841
Mingying Chuo UNC Institute of Marine Sciences 252 726 6841
Mary Kate Rinderie UNC Institute of Marine Sciences 252 726 6841

FWS Form 3-1383-R(Rev.03/2020) OMB Control No. 1018-0102
U.S.Department of the Interior Expires 01/31/2024
Project Information
11)Title of project: Proposed Monitoring Plan for Assessing the Efficacy of Peroxide Treatment of Cyanobacteria in Lake Mattamuskeet
12)Is a full research proposal attached? Yes O No O
Note: Depending on the research and monitoring project for which you are requesting a permit,we may ask you for the following
project information(13-25)if it is not included in your research proposal,or if you have not provided a full research proposal with
this application.Please contact the specific refuge where the activity is being conducted to determine what information is required.
Attach additional sheets to the application if the text spaces provided are inadequate.
13) Describe project by specifically identifying timing,frequency, and how the project is expected to proceed:
The purpose of this project is to assess the efficacy of treatments by Bluegreen Water Technologies to reduce levels of harmful cyanobacteria in Lake
Mattamuskeet.A monitoring program will be implemented within the lake to assess conditions before,during and after treatments.Our team from UNC
Chapel Hill will visit 20 stations on at least a monthly basis throughout the duration of the project. Sixteen of the 20 stations are located within 8 coves
that have been identified as either treatment areas that will receive peroxide or control areas that will not receive treatment. Each cove will have 2
sampling stations and there wit be 2 stations in the central portion of both the east and west lake basins.On each basin one of the central lake stations
will be co-located with instrumentation continuously operated by the USGS.At each sampling time point,our team will measure indicators of total
phytoplankton and cyanobacterial biomass including chlorophyll a,phycocyanin,and taxa-specific carotenoids.We will also measure dissolved and
particulate organic and inorganic nitrogen forms and dissolved inorganic and total phosphorus.Depth profiles of temperature,salinity,dissolved
oxygen,pH.turbidity,chlorophyll a fluorescence,and photosynthetically active radiation will also be measured at each sampling point. Monthly
monitoring will occur from November through February and twice monthly monitoring will occur from March through October. During the period
immediately before and after treatment,monitoring frequency will increase to daily for the first 2 days and then every other day for the next 10 days to
capture acute impacts of the treatment.During the treatment period,zooplankton biomass/community structure and cyanotoxins will additionally be
monitored.
14) Specifically identify location(s)and/or attach a map for the project: (GPS location(s)preferred)
See map and table in attached monitoring plan.
15) Identify species or habitats being studied:
All habitats being studied are open-water habitats.
16) Purpose/hypothesis:
-We hypothesize that the cyanobacterial treatment will reduce cyanobacterial biomass levels and
provide measurable improvement to water clarity. The degree of effectiveness and duration of
treatment effects will be carefully documented.
17) Expected benefits of research/monitoring:
The project will provide the first large-scale, controlled experimental assessment of peroxide treatment for decreasing
cyanobacteria biomass. The phytoplankton, nutrient, and optical data collected during the project will additionally provide
great benefit for understanding and managing Lake Mattamuskeet.
18) Briefly describe project history and context of research/monitoring project:
The NC State legislature passed a bit that funded a trial of peroxide treatment to reduce levels of harmful cyanobaderia biomass within a North Carolina water body.Lake Mattamuskeet was proposed
as a potential test site due to its consistently high levels of cyanobadena biomass and the hope of improving water clarity for restoring submerged aquatic vegetation by reducing cyanobacterial biomass.
The bill that funded the experimental treatment also funded our UNC team to provide a third party,independent assessment of the efficacy of the treatment and a documentation of any unintended
consequences.
19) Briefly describe project's relationship to other research/monitoring projects either known of or conducted by the applicant:
This project will complement on-going water quality monitoring efforts conducted by Lake Mattamuskeet NWR and the
USGS. It will significantly improve understanding of seasonality and spatial variability of phytoplankton and nutrients
within Lake Mattamuskeet

FWS Form 3-1383-R(Rev.03/2020) OMB Control No 1018-0102
U S Department of the Interior Expires 01/31/2024
20)Identify the types of specimen collections to be taken(see specimen collection clause in the instruction section#20)or data to be collected
during the proposed project
1) raw water samples
2) zooplankton net tow samples
21) List other cooperators and institutions involved in the project
t r Technologies Bluegreen Water e
22)Generally identify the anticipated timeline for analysis.write-up and publication:
Jan 2023 to Dec 2024-data collection
Jan 2025 to June 2025-data analysis and write up
Fall 2025-publication
23) For research involving animals, attach an Assurance of Animal Care Form or an approval from an Institutional Animal Care and Use
Committee?Is a form or approval attached? O Yes O No O N/A
License/Insurance/Certifications/Permits
Note: Contact the specific refuge office where the research project is going to be conducted to determine if any type of license,
insurance,certification(s),or permit(s)will be required.We may process this Special Use Permit while the applicant obtains them.
Attach additional sheets to the application if the text spaces provided are inadequate.
24a)List and attach copies of any licenses you have for equipment operation (i.e., aviation or commercial boats), pesticide applications,
transporters)or others if required:
License Type Number Expiration Date (if applicable)
N/A
24b)List and attach copies of any insurance you have(i.e general liability,flight/grounding,contaminants.medical evacuation,or others if required:
Insurance Type Carrier Expiration Date (If applicable)
N/A Blue Cross Blue Shied
24c) List and attach copies of any certifications you have, such as rat free, hull inspections, CPR/First Aid,or others if required:
Certificate Type Expiration Date(if applicable)
all UNC staff have medical insurance that covers medical evacuation
24d)List and attach copies of any other Federal, State,or Tribal permits if required:
Permit Type Permit Number Expiration Date (if applicable)
N/A
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FWS Form 3-1383-R(Rev.03/2020) OMB Control No. 1018-0102
U.S. Department of the Interior Expires 01/31/2024
Logistics and Transportation Attach additional sheets if the text spaces provided are inadequate.
25a)Does project require personnel to stay overnight on the refuge? Yes O No O
25b)If yes, how many? And list known personnel involved in overnight stay below.
List Names List Names List Names List Names
26)Specifically describe all major instrumentation/equipment/gear(i.e. use of drones)and materials used, if applicable or required:
We plan to install 8 In Situ multiparameter water quality instruments. One instrument will be located
within each of the 8 project study coves and will continuously measure pH, dissolved oxygen,
temperature and conductivity.
27a)Provide details and schedule for the installation of instrumentation:
We plan to install the instruments by hanging them horizontally below a float. Instruments will be
deployed in late February and will be coordinated with refuge management to minimize impacts to
waterfowl.
27b) Provide details and schedule for the removal of instrumentation:
Instrumentation will be removed December 2025.
27c) If instrumentation is permanent, describe need:
Instrumentation is not permanent.
27d) If instrumentation requires a maintenance schedule,describe needs and schedule:
nstrumentation will be cleaned and calibrated by UNC staff during scheduled sampling trips to Lake Mattamuskeet.
27e)Provide a data collection schedule:
In project years 1 and 2, we anticipate sampling monthly during November, December, January, and February and then
twice monthly from March through October. Exact sampling dates and times will be coordinated with refuge staff to
minimize impacts on waterfowl and other refuge uses (e.g. permitted duck hunts).
28)Provide logistical arrangements for offsite transportation of samples:
Samples will be transported by UNC staff to the laboratory at UNC's Institute of Marine Sciences.
29a)Provide detailed information on the logistics for onsite, intersite, and/or ship-to-shore transportation to or on the refuge, if required:
We plan to use a 17' foot double wide (low draft) Carolina skiff to visit sampling sites. We plan to
launch at the pump house canal ramp and the Rose Bay canal ramp.
Page 4 of 9

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FWS Form 3-1383-R(Rev. 03/2020) OMB Control No 1018-0102
U.S. Department of the Interior Expires 01/31/2024
29b)Provide descriptions, license plate and/or identification numbers of vehicles used for onsite transportation, if required:
Vehicle Type Plate/I.D./Registration# Vehicle Type Plate/I.D./Registration#
Chevrolet pick up truck NC plate#: 'PJ 4579"
Ford pick up truck NC plate#: "PJ 3951'
Chevrolet pick up truck NC plate#: "PA 4475'
Ford passenger van NC plate#: "PL 8104"
29c) Provide descriptions. license plate and/or identification numbers of vehicles used for intersite transportation, if required:
Vehicle Type Plate/I.D./Registration# Vehicle Type Plate/I.D./Registration#
Chevrolet pick up truck NC plate#: 'PJ 4579*
Ford pick up truck NC plate#: "PJ 3951'
Chevrolet pick up truck NC plate#: 'PA 4475
Ford passenger van NC plate#: "PL 8104"
29d)Provide descriptions,license plate and/or identification numbers of vehicles used for ship-to-shore transportation, if required
Vehicle Type Plate/I.D./Registration# Vehicle Type Plate/I.D./Registration#
N/A N/A
30a) Is fuel cache needed? Yes O No O N/A O 30b) Provide specific location(s)of fuel caches: (GPS coordinates preferred)
31)Attach Safety Plan if required. Is Safety Plan attached?Yes O No O N/A 0
Work and Living Accommodations
32) Specifically describe onsite work and/or living accommodations, including spike camps:
N/A
33) Specifically describe on or offsite hazardous material storage or other on or offsite material storage space:(Including on and offsite fuel caches
N/A
Sign,date,and print this form and return it to the refuge for processing.By signing this application,I agree my operations will
conform to the information I have provided in this application,and I understand that any deviations or changes to this information
must receive prior written approval.
Nathan Hall Digitally signed by Nathan Hall 3/4/2024
34)Signature of Applicant: Date:2024 o3.oa 1z:3s:37 osoo Date of Application:
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FWS Form 3-1383-R(Rev. 03/2020) OMB Control No 1018-0102
U S. Department of the Interior Expires 01/31/2024
NOTICES
No Members of Congress or Resident Commissioner shall participate in any part of this contract or to any benefit that may arise from it,but this
provision shall not pertain to this contract if made with a corporation for its general benefit.
The Permittee agrees to be bound by the equal opportunity"nondiscrimination in employment"clause of Executive Order 11246.
PRIVACY ACT STATEMENT
Authority: The information requested is authorized by the National Wildlife Refuge System Administration Act (16 U.S.C. 668dd-ee)and the
Refuge Recreation Act(16 U.S.C.460k-460k-4).
Purpose: To collect the applicant's information to process permits allowing for: research and monitoring activities by students, universities, or
other non-FWS organizations.
Routine Uses: The information will be used by the refuge's administrative office for processing Research and Monitoring Special Use permits.
More information about the routine uses maybe found in the Systems of Records Notice, FWS-5 National Wildlife Refuge Special Use Permits.
Disclosure: Providing the information is voluntary. However, submission of information is required to process and approve research and
monitoring activity usage on the National Wildlife Refuge System.
PAPERWORK REDUCTION ACT STATEMENT
We are collecting this information subject to the Paperwork Reduction Act(44 U.S.C. 3501)to evaluate the qualifications,determine eligibility,
and document permit applicants and to respond to requests made under the Freedom of Information Act and the Privacy Act of 1974. The
information that you provide is required to obtain or retain a benefit; however, failure to provide all required information is sufficient cause for the
U.S. Fish and Wildlife Service to deny a permit. False,fictitious, or fraudulent statements or representations made in the application may be
grounds for revocation of the Special Use Permit and may be punishable by fine or imprisonment(18 U.S.C. 1001). According to the
Paperwork Reduction Act of 1995, an agency may not conduct or sponsor and a person is not required to respond to a collection of
information unless it displays a currently valid OMB control number. OMB has approved this information collection and assigned control
number 1018-0102.
ESTIMATED BURDEN STATEMENT
The public reporting burden for this information collection varies based on the requested specific refuge use. We estimate the relevant public
reporting burden for the Commercial Activity Special Use Permit Application form(Research and Monitoring)to average 5 hours per response,
including the time for reviewing instructions, searching existing data sources, gathering and maintaining the data needed, and completing and
reviewing the collection of information. Send comments regarding this burden estimate or any other aspect of this collection of information,
including suggestions for reducing this burden,to the Service Information Clearance Officer, U.S. Fish and Wildlife Service, 5275 Leesburg
Pike, MS: PRB (JAO/3W), Falls Church,VA 22041-3803, or via email to Info Collfws.qov. Please do not send your completed form to this
address.
GENERAL CONDITIONS AND REQUIREMENTS
1)Responsibility of Permittee: We shall consider the permittee, by operating on the premises, to have accepted these premises with all
of the date of thispermit.At the end of theperiod specified or upon earlier
facilities, fixtures, or improvements in their existing condition as p p
termination, the permittee shall give up the premises in as good order and condition as when received except for reasonable wear,tear,or
damage occurring without fault or negligence. The permittee will fully repay the Service for any and all damage directly or indirectly resulting
from negligence or failure on his/her part, and/or the part of anyone of his/her associates,to use reasonable care.
2) Operating Rules and Laws: The permittee shall keep the premises in a neat and orderly condition at all times, and shall comply with all
municipal, county. and State laws applicable to the operations under the permit as well as all Federal laws, rules, and regulations governing
national wildlife refuges and the area described in this permit. The permittee shall comply with all instructions applicable to this permit issued
by the refuge official in charge. The permittee shall take all reasonable precautions to prevent the escape of fires and to suppress fires and
shall render all reasonable assistance in the suppression of refuge fires.
3) Use Limitations: The permittee's use of the described premises is limited to the purposes herein specified and does not, unless provided for
in this permit, allow him/her to restrict other authorized entry onto his/her area; and allows the U.S. Fish and Wildlife Service to carry on
whatever activities are necessary for: (1) protection and maintenance of the premises and adjacent lands administered by the U.S. Fish and
Wildlife Service; and (2)the management of wildlife and fish using the premises and other U.S. Fish and Wildlife Service lands.
4)Transfer of Privileges: This permit is not transferable, and no privileges herein mentioned may be sublet or made available to any person or
interest not mentioned in this permit. No interest hereunder may accrue through lien or be transferred to a third party without the approval of the
Regional Director of the U.S. Fish and Wildlife Service and the permit shall not be used for speculative purposes.
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5) Compliance: The U.S. Fish and Wildlife Service's failure to require strict compliance with any of this permit's terms, conditions, and
requirements shall not constitute a waiver or be considered as a giving up of the U.S. Fish and Wildlife Service's right to thereafter enforce any
of the permit's terms or conditions.
6) Conditions of Permit not Fulfilled: If the permittee fails to fulfill any of the conditions and requirements set forth herein,the U.S.Fish and Wildlife
Service shall retain all money paid under this permit to be used to satisfy as much of the permittee's obligation as possible.
7) Payments:All payment shall be made on or before the due date to the local representative of the U.S. Fish and Wildlife Service by a postal
money order or check made payable to the U.S. Fish and Wildlife Service.
8)Termination Policy: At the termination of this permit the permittee shall immediately give up possession to the U.S. Fish and Wildlife Service
representative, reserving, however, the rights specified in paragraph 11 below. If he/she fails to do so, he/she will pay the U.S. Fish and
Wildlife Service, as liquidated damages, an amount double the rate specified in this permit for the entire time possession is withheld. Upon
yielding possession,we will still allow the permittee to reenter as needed to remove his/her property as stated in paragraph 11 below.The
acceptance of any fee for the liquidated damages or any other act of administration relating to the continued tenancy is not to be considered as
an affirmation of the permittee's action nor shall it operate as a waiver of the U.S. Fish and Wildlife Service's right to terminate or cancel the
permit for the breach of any specified condition or requirement.
9) Revocation Policy: The Regional Director of the U.S. Fish and Wildlife Service may revoke this permit without notice for noncompliance with
the terms hereof, or for violation of general and/or specific laws or regulations governing national wildlife refuges, or for nonuse. It is at all times
subject to discretionary revocation by the Director of the Service. Upon such revocation the U.S. Fish and Wildlife Service, by and through any
authorized representative. may take possession of said premises for its own and sole use, and/or may enter and possess the premises as the
agent of the permittee and for his/her account.
10) Damages: The U.S. Fish and Wildlife Service shall not be responsible for: any loss or damage to property including but not limited to crops,
animals, and machinery; injury to the permittee or his/her relatives or to the officers. agents, employees, or any other(s)who are instructed to
be on the premises; the sufferance from wildlife or employees or representatives of the U.S. Fish and Wildlife Service carrying out their official
responsibilities. The permittee agrees to hold the U.S. Fish and Wildlife Service harmless from any and all claims for damages or losses that
may arise to be incident to the flooding of the premises resulting from any associated Government river and harbor, flood control, reclamation,
or Tennessee Valley Authority activity.
11) Removal of Permittee's Property: Upon the expiration or termination of this permit,if all rental charges and/or damage claims due to the U.S.
Fish and Wildlife Service have been paid.the permittee may,within a reasonable period as stated in the permit or as determined by the U.S. Fish
and Wildlife Service official in charge,but not to exceed 60 days, remove all structures, machinery, and/or equipment, etc., from the premises
for which he/she is responsible. Within this period the permittee also must remove any other of his/her property including his/her acknowledged
share of products or crops grown,cut, harvested, stored,or stacked on the premises. Upon failure to remove any of the above items within the
aforesaid period, they shall become the property of the U.S. Fish and Wildlife Service. .
INSTRUTIONS FOR COMPLETING APPLICATION
You may complete the application portion verbally, in person, or electronically and submit to the refuge for review. Note: Please read instructions
carefully as not all information is required for each activity. Contact the specific refuge where the activity will take place if you have questions
regarding the applicability of a particular item. We may add special conditions or permit stipulations to permit prior to approval.
to-1c) Identify if permit application is for new, renewal, or modification of an existing permit, whether or not you have or will be applying to
another refuge for the same activity, and for which refuge(s). Permit renewals may not need all information requested. Contact the specific
refuge headquarters office where the activity is going to be conducted to determine applicability of this requirement.
2-3) Provide principal investigator or applicant full name. Attach principal investigator's Curriculum Vitae or Resume, if required. Permit
renewals generally do not require a Curriculum Vitae or Resume if the project is a continuation of a previously issued permit being
conducted by the same investigator. Contact the specific refuge office to determine applicability of this requirement.
4-9) Provide investigator's physical and/or mailing address, phone, fax, e-mail, affiliation and/or sponsoring organization, and
relationship to affiliation or organization (title, professor, student,etc.).
10) Provide the names and addresses of assistants, subcontractors, or subpermittees. 1Ak may require names and addresses if the assistants,
subcontractors or subpermittees will be operating on the refuge without the permittee being present. Volunteers, assistants, subcontractors, or
subpermittees accompanied by the permittee need not be identified.
11) Provide title of research or monitoring project.
12a-12b) Attach a full research or monitoring proposal, if required. Permit renewals generally do not require a project proposal if the project is a
continuation of a previously issued permit being conducted by the same investigator. Contact the specific refuge office to determine applicability
of this requirement.
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1

FWS Form 3-1383-R(Rev.03/2020) OMB Control No 1018-0102
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13) Provide detailed information on the activity, including timing, frequency, how the project is expected to proceed, etc. Permit renewals may not
need activity description, if the activity is unchanged from previous permit. Most repetitive research projects do not require an activity description
for each visit to the refuge. Contact the specific refuge office to determine applicability of this requirement.
14) Identify specific location (GPS coordinates preferred) if not a named facility,and/orattach a map of the location. Permit renewals may not require
a location if the project is essentially unchanged from the previous permit. Contact the specific refuge office to determine applicability of this
requirement.
15) Identify species or habitats being studied.
16-17) Specifically identify purpose or hypothesis of the research or monitoring project and describe expected benefits. Permit renewals may
not need to identify purpose or hypothesis if the project is a continuation of a previously issued permit being conducted by the same
investigator. Contact the specific refuge office to determine applicability of this requirement.
18) Briefly describe project history and context. Permit renewals should describe previous research activities as part of a previously issued
permit being conducted by the same investigator.Contact the specific refuge office to determine applicability of this requirement.
19) Briefly describe project's relationship to other research/monitoring projects either known of or conducted by the applicant, if applicable.
Include a brief statement of how the research or monitoring permit being applied for will add to or supplement other ongoing research or
monitoring on the same, or related, species or habitats. Contact the specific refuge office to determine applicability of this requirement.
20) Identify specimen collections to be taken or types of data to be collected. You may use specimens collected under this permit, any
components of any specimens (including natural organisms, enzymes, genetic materials or seeds), and research results derived from collected
specimens for scientific or educational purposes only, and not for commercial purposes unless you have entered into a Cooperative Research and
Development Agreement (CRADA) with us. We prohibit the sale of collected research specimens or transfers to third parties for commercial
purposes. Breach of any of the terms of this permit will be grounds for revocation of this permit and denial of future permits. Furthermore. if you
sell or otherwise transfer for commercial purposes collected specimens. any components thereof, or any products or research results developed
from such specimens or their components without a CRADA, you will pay us a royalty rate of 20 percent of gross revenue from such sales. In
addition to such royalty,we may seek other damages and injunctive relief against you. Permit renewals may not need to identify samples taken if
the project is a continuation of a previously issued permit being conducted by the same investigator. Contact the specific refuge office to
determine applicability of this requirement.
21) List other cooperators and institutions involved in the project, if applicable. Contact the specific refuge office to determine applicability of this
requirement.
22) Generally, identifythe anticipated time line for analysis,write-up, and publication of project results. Include whether the project is a single. or
Y P Y
multiple year project. Identification of an actual publication where the results are printed is not necessary. However, applicants should include
the anticipated dissemination of project results. Contact the specific refuge office to determine applicability of this requirement.
23) Check box acknowledging a completed Assurance of Animal Care Form or an Institutional Animal Care and Use Committee (or
equivalent) that has granted approval has been completed,and has been submitted to refuge station,if required.Contact the specific refuge office
to determine applicability of this requirement.
24a-24d) Specifically identify types and numbers of licenses, insurance, certifications, and other State, Federal, or Tribal permits if required.
Contact the specific refuge headquarters office where the project is going to be conducted to determine applicability of these requirements,
and to coordinate the simultaneous applications of any of these requirements while this Special Use Permit is being processed.
25a-25b) Provide the number of and/or name(s)of any personnel required to stay overnight on the refuge, if applicable.
26) Identify all equipment(including drones) and materials that will be used, if required. Permit renewals may not require a list of equipment if
the project is essentially unchanged from a previously issued permit. Contact the specific refuge office to determine applicability of this
requirement.
27a-27e) Identify types and schedule(s) of installation of any instrumentation, data collection, and maintenance schedule of instrumentation, if
required. Permit renewals may not require a list of equipment if the project is essentially unchanged from a previously issued permit. However,
schedules of installation of any instrumentation, data collection, and maintenance schedule of instrumentation may still be required. Contact the
specific refuge headquarters office where the project is going to be conducted to determine applicability of this requirement.
28) Identify logistical arrangements for offsite transportation of samples taken, if applicable.
29a-29d) Describe and provide vehicle descriptions and license plate or identification numbers of all vehicles, including boats and airplanes, if
required. Motor vehicle descriptions are only required for permittee vehicle, and/or if the vehicle will be operated on the refuge without the
permittee being present. Motor vehicles that are accompanied by the permittee as part of a group (convoy) activity need not be identified if
Page 8 of 9

FWS Form 3-1383-R(Rev. 03/2020) OMB Control No 1018-0102
U S Department of the Interior Expires 01/31/2024
cleared in advance by refuge supervisor. Specifically describe ship-to-shore, intersite (between islands. camps, or other sites) and onsite
transportation mechanisms, and license plate or identification numbers, if required.
30a-30b)Identify specific location(s)of fuel cache(s) (GPS coordinates preferred), if required.
31 a-31 b)Attach safety plan, if required. Contact the specific refuge office to determine applicability of this requirement.
32) Specifically describe onsite work and/or living accommodations, if required. Include descriptions and locations (GPS coordinates preferred)
of spike camps or other remote work and/or living accommodations that are not part of the base of operations. Contact the specific refuge
office to determine applicability of this requirement.
33) Specifically describe onsite and offsite hazardous material storage, or other onsite material storage space (including on and offsite fuel
caches), if required. Contact the specific refuge office to determine if descriptions of hazardous material storage or other onsite material
storage are required.
34) Sign, date, and print the application. Click on the Print button to print the application (if using the Tillable version). The refuge official will
review and, if approved, fill out the remaining information, sign, and return a copy to you for signature and acceptance.
THIS APPLICATION FORM IS NOT VALID AS A PERMIT
BUT MAY BE USED AS A REFERENCE DOCUMENT ATTACHED TO THE OFFICIAL PERMIT.
ONLY OFFICIAL REFUGE PERSONNEL MAY ASSIGN A VALID PERMIT NUMBER AND PERMIT TERM
TO THIS APPLICATION FORM AFTER THE PERMIT HAS BEEN APPROVED.
Page 9 of 9

RECEIVED
APR 12 2024
Nathan Hall and Hans Paerl
UNC Institute of Marine Sciences NCDEQ/DWR/NPDES
2/19/2024
Monitoring Plan for Assessing the Efficacy of Peroxide Treatment of Cyanobacteria
in Lake Mattamuskeet
Currently, Lake Mattamuskeet is hypereutrophic with an average chlorophyll a concentration of 95 µg/L,
nearly 2.5 times the 40 ug/L NC State water quality standard.The phytoplankton community is
dominated by thin filamentous cyanobacteria that do not produce highly visual surface scums.The
primary use impairments associated with the excessive cyanobacteria biomass in Lake Mattamuskeet
are decreased light penetration for submerged aquatic vegetation (SAV) (Moorman et al. 2017) and
toxin production that may accumulate in blue crabs that are fished for human consumption (Moorman
2018). Experimental treatments of isolated areas on both the east and westerns sides (east and west of
HWY 94)of Lake Mattamuskeet are proposed to reduce cyanobacteria biomass to levels, improve water
clarity, and reduce cyanotoxins. We expect that treatments will occur during late spring/early summer
of 2024. From January 2023 through February 2024,we have monitored the phytoplankton community,
nutrient levels,and optical water quality constituents prior to the treatment to establish a robust
baseline. Intensive monitoring prior to the treatment will include additional parameters aimed at
quantifying and characterizing the zooplankton community and cyanobacterial toxin levels. Intensive
monitoring immediately following the treatment will capture rapid changes in the phytoplankton
assemblage,toxin levels, zooplankton community, and water clarity that result from the treatment but
will also capture any negative side effects such as drops in dissolved oxygen or acute changes in
zooplankton community structure. Monitoring will continue, on a less intensive basis,for the remainder
of the year following treatment to quantify the longevity of the treatment's
impact on reducing cyanobacteria and to document other longer-term Table 1.Sampling schedule
changes in conditions (e.g. improved water clarity). Four treatment areas Month #Sampling
and four control sites will be equally distributed between the east and west Trips
sides of the lake and will be isolated from the rest of the lake using turbidity Jan 1
curtains. By comparing before and after and the inclusion of replicated Feb 1
control and treatment areas,this project design fulfills the design Mar 2
requirements for a before/after/control/interrupted (BACI) experimental Apr 2
design,the "gold standard"for environmental experiments. May(treat) 8
Jun 2
Monitoring Schedule Jul 2
During 2023, monitoring occurred approximately monthly during the winter Aug 2
months (Dec-Mar) and the frequency was increased to approximately twice Sep 2
monthly during spring through fall. We plan to keep this schedule during Oct 1
Nov 1
2024 except that we plan to conduct high frequency, approximately daily, Dec 1
monitoring during the week following treatment in late spring early
summer 2024.
Monitoring sites
Eight project areas will be assigned as four control and four treatment areas with two controls and two
treatment areas on each side of the lake (Figure 1).Two monitoring stations have been established

1
within each project area. In between each area's two monitoring stations, an In Situ Aqualtroll 600
water quality instrument has been deployed with continuous monitoring capabilities for temperature,
conductivity, pH,and dissolved oxygen (see continuous monitoring description below).Additionally,
Bluegreen has equipped each area with an AquaRealTime probe that continuously measures turbidity,
and in vivo fluorescence of chlorophyll a and phycocyanin. Both stations will be sampled on each
sampling trip to provide duplicate observations from each area. Representativeness of these project
areas will be gaged by comparison against four main lake stations that will include the USGS real time
monitoring locations in both the east (EM) and west(WM) sides of Lake Mattamuskeet(Figure 1).
x
aaayyt. Fa q b
1£ast Treatmen4E,BA
F' i
l,East Control B
•
-° I jEasr Corr,,
West Treetrpent A
-„ os°.,.,,>,•x>,.a r.:. East Treatment A
West Treatment B ,West Control B
West Control Ar / •
Lailo
Figure 1. Map showing the planned set of four treatment and four control project areas to assess
treatment efficacy. Red lines outline the project areas and white lines indicate the turbidity curtains that
will isolate these areas from the rest of the lake. Representativeness of these project areas will be gaged
by comparison against four main lake stations that will include the USGS real time monitoring locations
in both the east(EM)and west(WM)sides of Lake Mattamuskeet.
Parameters: Discrete sampling, unattended continuous data collection, and remote sensing will be used
to measure a broad range of water quality and habitat characteristics to provide a comprehensive
assessment of the treatment impacts on the phytoplankton community, and habitat suitability for fish
and submerged aquatic vegetation in Lake Mattamuskeet.Table 2 shows a list of proposed parameters
and their utility for the project.
Continuous measurements:Aqua Real Time Algae Tracker © continuous in vivo fluorescence sensors
are deployed within the project areas and the main lake by Blue Green Water Technologies to measure
total phytoplankton biomass as chlorophyll a fluorescence, cyanobacterial biomass as phycocyanin
fluorescence, and suspended sediments as turbidity. Data from these sensors are available to UNC-IMS
via an online dashboard. Dissolved oxygen, pH, temperature,and salinity is being monitored
continuously at one centralized station within each of the project areas using an In situ Aqua Troll 600
multiparameter sonde.The USGS real time instruments will continuously monitor pH, dissolved oxygen,

temperature, and salinity within the main lake. Continuous dissolved oxygen and pH measurements will
be used to evaluate any potential negative side effects such as sags in dissolved oxygen or pH that result
from the mortality of significant levels of cyanobacterial biomass.Temperature and salinity data will be
used as fundamental habitat information for phytoplankton, SAV, and fish, and also to help evaluate
circulation features during the course of the study. For example,salinity intrusions from Pamlico Sound
may cause changes in cyanobacteria biomass and monitoring salinity will allow those events to be
detected.Additionally, spatial gradients in salinity may be used to indicate the degree to which turbidity
curtains are able to isolate treatment and control project areas from the surrounding lake water.
Discrete sampling:As described above, a total of 20 stations for discrete sampling have been
established with 2 stations in each of the 8 project areas and 2 stations in each of the east and west
sides of the main lake area (Figure 2).
•
•
.'
r
Figure 2. Map showing the planned 20 sampling stations within the project areas outlined in red/white
lines and the main lake.

1
Table 2,Coordinates for 20 planned sampling
locations
Discrete sampling stations are co-located with Station Number 'latitude 'Longitude
unattended monitoring instrumentation to provide the 3 35 5132 -76 2585
maximum capacity for data comparisons and post 4 35 4829 -76 2725
calibration of the continuous data streams. A total of 5 35 4832 .76 2699
25 sampling trips are planned for each project year to 6 35 4750 76 2714
7 35 4698 -76 2691
provide adequate information on pre and post 8 35 4733 .76 25,74
treatment conditions in the treatment, control, and 9 35 4766 -76 2599
main lake areas.The temporal frequency of sampling I 11 35 4773 76 2495
trips throughout the year will be varied to achieve 12 35 4768 76 2499
13 35 5056 -76 2537
higher resolution during the period when treatments 22 35 5064 •76 1839
occur and during summer when cyanobacterial bloom 26 35 5106 -76 1313
activity is most likely(Table 1). 29 35 4878 -76 1043
30 35 4892 -76 0988
On each sampling event, measurements of chlorophyll 31 35 5041 -76 0755
a and accessory pigments determined via high pressure 32 35 5016 76 0733
35 35 5160 .76 0715
liquid chromatography and phycocyanin via in vitro 36 38 5135 -76 0711
fluorometry are providing a robust estimation of the 37 35 5223 -76 0740
total and class-level changes in phytoplankton biomass 20 35 5217 -76 0739
emphasizing cyanobacteria. An aliquot of each sample
is preserved in 1%Lugol's iodine and selected samples are microscopically examined to assess changes
in the abundance of known toxin producers (e.g. Cylindrospermopsis raciborskii)and determine how the
abundance of toxin producers relates to measurements of cyanotoxins. Dissolved and particulate
nitrogen and phosphorus concentrations (nitrate+nitrite, ammonium,total dissolved nitrogen, dissolved
organic nitrogen, particulate nitrogen, o-phosphate, and total P) are measured from every sample to
provide information on how the peroxide treatment affects dynamics between the phytoplankton and
nutrient pools.These nutrient data will be critical for understanding changes in community composition,
and the efficacy of the treatment in achieving long-term water quality improvements.
On each sampling event, depth profiles of chlorophyll a and phycocyanin fluorescence,turbidity, pH,
dissolved oxygen, conductivity, and temperature are measured using a YSI 6600 multiparameter data
sonde. Depth profile data will be used to understand the vertical distribution of these parameters and
the near surface values will be compared against the continuous, real-time data collected by the Algae
Tracker and Aqua Troll 600 instruments as a quality assurance measure.Vertical profiles of
photosynthetically active radiation (PAR) are measured to quantify how changes in the phytoplankton
community impact light availability for SAV. Chlorophyll a,turbidity and colored dissolved organic
matter are the primary indicators for light attenuating substances in water.These substances are at
each sampling event to enable modeling PAR attenuation to tease apart their contributions to PAR
attenuation and isolate the impact due to changes in cyanobacterial abundance.
Zooplankton biomass and community structure will be monitored during the period immediately before
and after treatments to assess positive or negative impacts to the zooplankton community which
supports the lake's fishery. Similarly,cylindrospermopsin, microcystin, and BMAA cyanotoxins will be
measured from discrete sample collected prior to and immediately after treatment.
Remote sensing:
1

Blue Green Water Technologies plans to use remote sensing to provide data necessary for determining
their treatment strategy.They have agreed to share their remotely sensed data products with the UNC-
IMS research team.The UNC-IMS team will evaluate the remotely sensed data products and will use
those data as additional information on treatment efficacy if we feel the data strengthen our
assessment capacity.
Table 3. Parameter list
Parameters Assessment Purpose
Chlorophyll a Total phytoplankton biomass
Phycocyanin Cyanobacterial biomass
Accessory pigments by HPLC Taxa-specific phytoplankton biomass
Phytoplankton species-microscopy Cyanobacterial species
Chlorophyll a fluorescence Continuous total phytoplankton
Phycocyanin fluorescence Continuous cyanobacteria biomass
Nutrients (N and P) Limit algal growth
Cylindrospermopsin Primary cyanotoxin in Lake Mattamuskeet
Microcystin Most common cyanotoxin in NC coastal waters
BMAA Emergent toxin of concern in eastern NC
Dissolved oxygen Fish habitat
pH Fish habitat
Zooplankton biomass/community structure Fish habitat/ecosystem health
Turbidity Light availability for SAV
Colored dissolved organic matter Light availability for SAV
PAR attenuation Light availability for SAV
Salinity Fundamental habitat information
Temperature Fundamental habitat information

i1
Research & Monitoring
l Special Use Permit
Station #: 42530 (For Official Use Only) Permit #: R23-001
Permit Term: From 12/12/2022 To 12/31/2024
1) Principal Investigator Name/Affiliation: BlueGreen Water Technologies
2) Permit Status:a)Approved: O If approved, provide special conditions(if any)in the text box below.
b) Denied: ° If denied, provide justification in the text box below.
[Type in additional Special Conditions or Justification for Denied Permit in the space provided]
General and Standard Conditions are listed on the following pages.Additional conditions specific to this permit are as follows:
1.The permittee must coordinate all activities with refuge personnel to avoid conflicts with other management activities such as carp removal efforts,
invasive species control,etc. During carp removal efforts,the permittee will be required to coordinate with the carp removal team to remove any
probes that may interfere with netting efforts or other carp removal activities.
2. Boats may only be launched from the three improved boat ramps on the refuge or from private launch sites.No unimproved sites may be used.
3.During the lake closure period from November 1 to February 28,the following conditions apply:
a.Boat access is limited to one daily visit per month on each basin of the lake.
b.Refuge personnel must be notified 48 hours prior to visits and reserve the right to postpone visits based on conflicting activities.
c.Refuge personnel(1)must be allowed the opportunity to accompany the permittee or their contractor during these visits.
d.Visits on the east basin may not occur during the refuge waterfowl hunts.
3)Are there additional special conditions attached to the permit? Yes O No °
4)Are licenses/permits required, and have they been verified? Yes ° No iO N/A iO
5)Are Insurance and/or Certification(s) required, and have they been verified? Yes° No ° N/A O
6) Is an Assurance of Animal Care or Institutional Animal Approval form needed? Yes ° No ° N/A O
If yes, is the form attached? Yes O No O
7) Has a Minimum Requirements Decision Assessment been conducted? Yes°No°N/A°
If yes,is assessment attached? Yes O No O
8) Record of Payments: Exempt Q Partial ° Full °
Amount of payment: Record of partial payment:
9) Is a surety bond or security deposit required? Yes ° No ° N/A O
This permit is issued by the U.S.Fish and Wildlife Service and accepted by the applicant signed below,subject to the terms,covenants,
obligations,and reservations,expressed or implied herein,and to the notice,conditions,and requirements included or attached.A
copy of this permit should be kept on-hand so that it may be shown at any time to any refuge staff.
10) Permit approved/issued by:(Signature and title) 11) Permit accepted by:(si nature of permit )
KENDALL SMITH Digitally OD6-05'SMITHALL '
Date: 12/9/2022 Date: 12.08.2022

r
General Conditions and Requirements
1)Responsibility of Permittee:The permittee, by operating on the premises, shall be considered to have accepted these premises with
all facilities,fixtures,or improvements in their existing condition as of the date of this permit.At the end of the period specified or upon
earlier termination,the permittee shall give up the premises in as good order and condition as when received except for reasonable
wear, tear, or damage occurring without fault or negligence. The perm all permittee will fully repay the Service for any and damage g directly or
indirectly resulting from negligence or failure on his/her part,and/or the part of anyone of his/her associates,to use reasonable care.
2)Operating Rules and Laws:The permittee shall keep the premises in a neat and orderly condition at all times, and shall comply with
all municipal county, and State laws applicable to the operations under the permit as well as all Federal laws, rules,and regulations
governing national wildlife refuges and the area described in this permit.The permittee shall comply with all instructions applicable to
this permit issued by the refuge official in charge.The permittee shall take all reasonable precautions to prevent the escape of fires and
to suppress fires and shall render all reasonable assistance in the suppression of refuge fires.
3)Use Limitations:The permittee's use of the described premises is limited to the purposes herein specified and does not, unless
provided for in this permit.allow him/her to restrict other authorized entry onto his/her area; and allows the U.S. Fish and Wildlife
Service to carry on whatever activities are necessary for:(1)protection and maintenance of the premises and adjacent lands
administered by the U.S. Fish and Wildlife Service;and(2)the management of wildlife and fish using the premises and other U.S. Fish
and Wildlife Service lands.
4)Transfer of Privileges:This permit is not transferable, and no privileges herein mentioned may be sublet or made available to any
person or interest not mentioned in this permit. No interest hereunder may accrue through lien or be transferred to a third party without
the approval of the Regional Director of the U.S. Fish and Wildlife Service and the permit shall not be used for speculative purposes.
5)Compliance: The U.S. Fish and Wildlife Service's failure to require strict compliance with any of this permit's terms,conditions, and
requirements shall not constitute a waiver or be considered as a giving up of the U.S. Fish and Wildlife Service's right to thereafter
enforce any of the permit's terms or conditions.
6)Conditions of Permit not Fulfilled: If the permittee fails to fulfill any of the conditions and requirements set forth herein,the U.S. Fish
and Wildlife Service shall retain all money paid under this permit to be used to satisfy as much of the permittee's obligation as possible.
7) Payments:All payment shall be made on or before the due date to the local representative of the U.S. Fish and Wildlife Service by a
postal money order or check made payable to the U.S. Fish and Wildlife Service.
8)Termination Policy:At the termination of this permit the permittee shall immediately give up possession to the U.S. Fish and Wildlife
Service representative, reserving, however,the rights specified in paragraph 11 below. If he/she fails to do so, he/she will pay the U.S.
Fish and Wildlife Service, as liquidated damages, an amount double the rate specified in this permit for the entire time possession is
withheld. Upon yielding possession,the permittee will still be allowed to reenter as needed to remove his/her property as stated in
paragraph 11 below.The acceptance of any fee for the liquidated damages or any other act of administration relating to the continued
tenancy is not to be considered as an affirmation of the permittee's action nor shall it operate as a waiver of the U.S. Fish and Wildlife
Service's right to terminate or cancel the permit for the breach of any specified condition or requirement.
9)Revocation Policy: The Regional Director of the U.S. Fish and Wildlife Service may revoke this permit without notice for
noncompliance with the terms hereof,or for violation of general and/or specific laws or regulations governing national wildlife refuges,
or for nonuse. It is at all times subject to discretionary revocation by the Director of the Service. Upon such revocation the U.S. Fish and
Wildlife Service, by and through any authorized representative, may take possession of said premises for its own and sole use, and/or
may enter and possess the premises as the agent of the permittee and for his/her account.
10) Damages: The U.S. Fish and Wildlife Service shall not be responsible for: any loss or damage to property including but not limited
to crops,animals,and machinery; injury to the permittee or his/her relatives or to the officers, agents, employees,or any other(s)who
are on the premises from instructions;the sufferance from wildlife or employees or representatives of the U.S. Fish and Wildlife Service
carrying out their official responsibilities.The permittee agrees to hold the U.S. Fish and Wildlife Service harmless from any and all
claims for damages or losses that may arise to be incident to the flooding of the premises resulting from any associated government
river and harbor, flood control, reclamation,or Tennessee Valley Authority activity.
11)Removal of Permittee's Property: Upon the expiration or termination of this permit, if all rental charges and/or damage claims due to
the U.S. Fish and Wildlife Service have been paid,the permittee may,within a reasonable period as stated in the permit or as
determined by the U.S. Fish and Wildlife Service official in charge, but not to exceed 60 days, remove all structures, machinery, and/or
equipment,etc.,from the premises for which he/she is responsible.Within this period the permittee also must remove any other of
his/her property including his/her acknowledged share of products or crops grown,cut, harvested, stored,or stacked on the premises.
Upon failure to remove any of the above items within the aforesaid period, they shall become the property of the U.S. Fish and Wildlife
Service.

1

. ."mrgiV*
Coastal NC National Wildlife Refuge Complex:
Alligator River, Cedar Island, Currituck, Mackay Island,
Mattamuskeet, Pea Island, Pocosin Lakes, Roanoke River,
and Swanquarter National Wildlife Refuges
CONDITIONS FOR SPECIAL USE PERMITS&AUTHORIZED ACTIVITIES
1. The Permittee or authorized person,and all officers, agents,employees, representatives,and
clients of the Permittee or authorized person,shall comply with all Refuge, Federal,State,and
local regulations and conditions that apply to the special use activity. Failure to comply with any
applicable regulation or condition,and all Federal laws, rules,and regulations governing National
Wildlife Refuges and the area described in the Special Use Permit(Permit)or authorizing
documents may result in revocation of the Permit or authorized activity and/or criminal
prosecution. The Permit is non-transferable.
2. While engaged in a special use activity on the Refuge,the Permittee or authorized person,and
his/her officers, agents,employees,or representatives, shall be in possession of a cony of the
Permit or authorizing documents(including all attachments that contain conditions)and shall,
upon request by an authorized Refuge official or by any authorized local,state,or federal law
enforcement officer,display the copy authorizing their presence and activity on the Refuge and
shall furnish any other licenses and identification documents as may be requested.
3. Any accident that results in a personal injury(i.e.,an accident that requires professional medical
treatment)shall be reported to the Refuge Manager within 24 hours of the accident.
4. Entryon the Refuge during nighttime hours(i.e.,Y2-hour after sunset to%-hour before
sunrise)is prohibited, unless authorized in writing by the Refuge Manager.Access to areas
closed to the public is not allowed, unless authorized in writing by the Refuge Manager.
5. Vehicle travel shall only on be designated roads or routes of travel,unless authorized in writing
g
by the Refuge Manager.Vehicles, boats,trailers,and other equipment shall be parked in such a
manner that roads and trails.including canoe trails and navigable waters.are not to be
blocked. Roads and trails need to be accessible to other Refuge visitors and to Refuge staff,
cooperative farmers,fire trucks,emergency vehicles,maintenance equipment,and law
enforcement patrols.
6. Vehicles with catalytic converters shall be restricted to paved roads,recently maintained gravel or
dirt roads,or bare soil areas because of the high fire potential. Vehicles with catalytic converters
shall not be parked over high vegetation or other fire hazardous materials.
1 RF-001(Revised 01/26/2020)

7. All locked Refuge gates shall be closed and locked upon entering and leaving Refuge property.
Refuge gates should be left the way they are found(i.e.,open,closed,either locked or
unlocked as the case may be). All signs describing authorized and unauthorized entry through
a gate shall be followed unless entry is allowed by this permit.
8. Refuge gate or building keys shall not be loaned to other agencies,companies,or persons.If
there is a need for access by other persons, please have them contact the Refuge Manager.All
i
Refuge keys shall be returned to the Refuge Manager,or a designated staff member, within 10
calendar days, after expiration or termination of the Permit or authorized activity.
9. All dogs(or any other pet) must be confined or on a leash while on the Refuge, unless authorized
jn writing by the Refuse Manager. Leashed pets must be under the immediate control of the
Permittee or authorized person, or the leash must be secured to a stationary object.The leash
shall not be in excess of 10 feet in length. Dogs are not allowed in areas where dogs are
prohibited, unless authorized in writing by the Refuge Manager.
10. Possession or use of firearms,air guns, bows and arrows,cross bows, spears,or gigs;or illegal
knives,weapons,or devices;or explosives of any type is prohibited on the Refuge when engaged
in a special use activity,unless authorized in writing by the RefugeManager.
11. Littering is prohibited.gjl materials brought into the Refuge shall be removed and properly
disposed.Drink cans, bottles,candy wrappers,toilet paper,and other garbage and refuse shall
not be left on the Refuge.
12. The Refuge Manager shall be contacted before any surface work is done.This includes
mowing, road or trail improvements,digging,clearing or trimming of brush or vegetation,
installation of structures,etc.
13. The use of herbicides and pesticides on Refuge property is prohibited, unless authorized in
writing by the Refuge Manager. Unrestricted,over-the-counter-type, insect repellents may be
used on or near the body and clothing to repel biting or stinging insects.
14. No permanent or semi-permanent markings shall be made on any Refuge building,structure,
gate, post,sign,fence,tree,vegetation, or soil by either marking, painting,cutting,scratching,
blazing, mowing,digging,or other destructive method, unless authorized in writing by the
Refuse Manager. When needed,only temporary, removable markers(e.g.,flagging tape, survey
stakes, metal/paper/plastic tags,etc.)shall be used to mark site locations, plots,etc.Safety signs,
informational signs,and any other signs required by law or regulation for the special use activity
being conducted,shall be posted as required,but only with prior authorization by the Refuge
2 RF-001 (Revised 01/26/2020)

Manager.All markers and signs shall be removed upon conclusion of the special use activity or
upon expiration or termination of the Permit.
15. No permanent or semi-permanent fences, buildings, shelters,docks, piers,or other structures
or facilities may be erected, built,or placed on the Refuge,unless authorized in writing by the
Refuge Manager. No machinery,equipment, supplies, or materials may be placed or stored on
the refuge, unless authorized in writing by the Refuge Manager.
16. All open fires are prohibited, unless authorized in writing by the Refuge Manager. Leaving an
authorized open fire unattended or not completely extinguished is prohibited. Setting on fire or
causing to be set on fire any timber, brush,grass,or other inflammable material, including camp
or cooking fires, is prohibited, unless authorized in writing by the Refuge Manager. The use of
cutting torches, arc welders, or any other open flame/sparking devices(which are required to
conduct the special use activity)shall be exercised with caution and only with prior authorization
from the Refuge Manager or Refuge Fire Management Officer. When use of these devices is
necessary,the operator(s)shall have immediate access to appropriate fire control eauioment
(e.g.,fire extinguishers,shovels,etc.)and immediate communication access to local emergency
services(e.g.,cellular telephone,two-wayradio, etc.). Tobacco smokers shall practice caution
P
when smoking;shall completely extinguish all matches,cigars,cigarettes, and pipes;and shall
dispose of same in a proper container(e.g.,a vehicle ashtray).
17. The Refuge Manager should be contacted immediately at(252)473-1132 upon discovery of
any wildfire,or any leak,spill,or break in a pipeline,power line,canal,or dike,or any other
accident or incident that has the potential to have an adverse impact on the soil,wildlife,or
plants in the area.Any unusual wildlife sightings or suspected illegal activities should be
reported to the Refuge Manager.
18. It is unlawful to disturb,destroy, injure,collect, or take any wildlife, plant, natural object,
mineral, cultural or historical feature,or public property on the Refuge,unless authorized in
federal collections or scientific use permit
writing by the Refuge Manager. State and ed
regulations and conditions also apply.
19. Wildlife shall not be harmed or harassed and disturbance shall be kept to a minimum,this
includes all snakes(poisonous and non-poisonous snakes),unless authorized in writing by the
Refuge Manager.State and federal collections or scientific use permit regulations and
conditions also apply.
20. Any research or monitoring activities involving ground disturbance may require historic
preservation consultation with the Regional Office and/or State Historic Preservation Office.
Researchers must obtain an Archeological Resource Protection Act permit from the Regional
Office prior to obtaining a special use permit from the refuge.
3 RF-001 (Revised 01/26/2020)

21. All researchers will be required to obtain appropriate State and Federal permits for the capture
and possession of protected species,for conducting regulated activities in wetlands, and for
other regulated activities. Researchers may also need to provide an assurance of animal care
form or an institutional animal approval form, if applicable.
22. Research collection activities must adhere to the Service's policy regarding disposition of biotic
specimens:
i. You may use specimens collected under this permit,any components of any specimens
(including natural organisms,enzymes,genetic materials or seeds),and research results
( g g Y
derived from collected specimens for scientific or educational purposes only,and not
for commercial purposes unless you have entered into a Cooperative Research and
Development Agreement(CRADA)with us. We prohibit the sale of collected research
P g
specimens or transfers to third parties for commercial purposes. Breach of any of the
terms of this permit will be grounds for revocation of this permit and denial of future
permits. Furthermore, if you sell or otherwise transfer for commercial purposes
collected specimens,any components thereof, or any products or research results
developed from such specimens or their components without a CRADA,you will pay us
a royalty rate of 20 percent of gross revenue from such sales. In addition to such royalty,
we may seek other damages and injunctive relief against you(USFWS 2016). U.S. Fish
and Wildlife Service. 2016. Director's Order No. 109,Amendment 11: Use of Specimens
Collected on Fish and Wildlife Lands. March 31, 2016.
23. The use or possession of traps, snares,or other passive (i.e., unattended)collection devices,
which are used to collect wildlife, is prohibited, unless authorized in writing by the Refuge
Manager. Each individual trap,snare,or passive collection device shall have a weather-resistant,
permanent tag attached with the Permittee's,authorized person's,and/or organization's name
legibly marked on the tag 2Lshall have the Permittee's,authorized person's,and/or
organization's name legibly marked, imprinted,or engraved on the trap,snare,ordevice.
24. For protection of threatened,endangered, rare, sensitive or commercially desirable species,
publication of specific collection locations(latitude/longitude) is prohibited.The Refuge Manager
(or designee)must approve the release of this information. Photographic materials
(photographs,digital images and videos)that document activities involving threatened and
endangered species permitted by this Permit(e.g., handling wildlife,entering refuge during
closed hours,collecting of any kind,entering a closed area,disturbing habitat, etc.)may not be
posted in any public forum (Internet, Facebook, HerpNation,etc.)unless authorized in writing by
the Refuse Manager and will result in the revocation of this Permit.
4 RF-001 (Revised 0 1/2 612 02 0)

25. Permittee may use photographic materials in official (University/public agency)publications,
including the Internet but must include a statement that the activity was conducted under the
provision of a Special Use Permit.All data collected on refuge lands will be shared with the U.S.
Fish and Wildlife Service.
26. Recreational and commercial uses of unmanned aerial vehicles(drones)are prohibited on the
refuge. Permitted research projects using UAS technology will be considered using Federal
Aviation Administration (FAA),Office of Aviation Services(OAS),and Department of Interior(DOI)
Aviation Policy guidelines.
27. All researchers are required to submit a final report to the refuge upon completion of their work.
If the study is long-term,an interim progress report will be required. Failure to provide progress
reports and/or final reports could result in the revocation of the current permit or jeopardize the
issuance of future permits.
28. Researchers who publish the work in peer-reviewed publications are to provide copies to the
refuge.All reports, presentations, posters,articles or other publications will acknowledge the
Refuge System and appropriate refuges as partners in the research, provided that the Service
does not otherwise deem it appropriate to issue a disclaimer.The acknowledgement recognizes
that the research could not have been conducted without the existence of the refuge and its
support and cooperation.The research organization/agency or personnel in conjunction with the
Service will retain the use and ownership of all data/reports.
5 RF-001(Revised 01126/2020)

FWS Form 3-1383-R(Rev. 03/2020) OMB Control No. 1018-0102
U.S. Department of the Interior Expires 01/31/2024
M.� RESEARCH AND MONITORING
f< SPECIAL USE PERMIT
�,,��;1• National Wildlife Refuge System
Refuge Name: For Official Use Only
v
Address: 85 Mattamuskeet Rd. Swanquarter, NC 27885 -approved Permit#:
Attn:(Refuge Official) Kendall Smith `>tation#:
E-Mail: [e-mail] >ermit Term:from
Phone#: [phone]
Note:We do not require ali information for each Research project.See instructions at the end of the notice and contact the refuge
to determine applicability of a particular item. Attach additional sheets if the text spaces provided are inadequate.
1 a) Identify the type of Permit you are applying for: New O Renewal O Modification O Other O
1 b)Have you applied, or do you intend to apply, to any other refuges for this same activity? O Yes O No
1 c)If yes,which refuges?
Applicant Information
2) Principal investigator: Jessica Frost 3) Is curriculum vitae or resume attached? Yes O No O
4a)Affiliation/Sponsoring Organization: BlueGreen Water Technologies/University of North Carolina (UNC)at Chapel Hill
4b)Relationship to affiliation/sponsoring organization(professor, staff, student,etc.): Scientific Director
5)Physical Address: 301 South Hills Village Ste LL200 #452
City/State/Zip: Pittsburgh, PA 15241
6)Mailing Address: (if different than above)
City/State/Zip:
7)Phone# [phone]) Fax#: 9) E-mail: [e-mail]
10)List known assistants/subcontractors/subpermlttees: (Only required if the assistants/subcontractors/subpermittees wit be operating on the refuge without
the permittee being present)
Name Address Phone#
Moshe Haret 2510 47th Street#209/Boulder/Colorado/80301 .972(0)52 3455 546
Mike Pelz Asheville,North Carolina 8283845146
Hans Paerl UNC Chapel Hill Institute of Marine Sciences/3431 Arendell Street/Morehead City/North Carolina/28557 2527266841 ext.133
Nathan Hall UNC Chapel Hill Institute of Marine Soences/3431 Arendell Street/Morehead City/North Carolina/28557 2527266841 ext.228
Chris Lee 2510 47th Street#209/BoulderlColorado/80301 3033357855
Trent Nelson 11011 Longistics Way#100/Raleigh/NC/27610 3364422680

FWS Form 3-1383-R(Rev.03/2020) OMB Control No 1018-0102
U.S.Department of the Interior Expires 01/31/2024
Project Information
11)Title of project: In-Situ Water Quality Monitoring
12) Is a full research proposal attached? Yes O No O
Note: Depending on the research and monitoring proje which you are requesting a permit,we may ask you for the following
project information(13-25)if it is not included in your research proposal,or if you have not provided a full research proposal with
this application.Please contact the specific refuge where the activity is being conducted to determine what information is required.
Attach additional sheets to the application if the text spaces provided are inadequate.
13) Describe project by specifically identifying timing,frequency,and how the project is expected to proceed:
They will use 30-50 floating autonomous water probe devices which are minimally invasive and measure
six key water variables (air temperature, water temperature, turbidity, chlorophyll-a, phycocyanin, ambient
light), GPS coordinates, and reports data to the science teams. It is solar powered, cellular connected,
and has self-cleaning sensors that collect data 24/7, updating every 30 minutes. Each probe device will
be visually checked and cleaned for any biofouling every two months or as needed per the device's
internal maintenance alert warning system. The duration of deployment will range 12-24 months,
dependent on quality of data collected and agreement among the Mattamuskeet National Wildlife Refuge,
the UNC and site stakeholders.
14) Specifically identify location(s)and/or attach a map for the project: (GPS location(s)preferred)
30-50 floating autonomous water probe devices will be temporarily deployed at strategic locations across Lake
• Mattamuskeet. BlueGreen will consult and come to an agreement with Mattamuskeet National Wildlife Refuge on
placement of probe devices.
15) Identify species or habitats being studied:
Data collected from all probe devices will contribute to the creation of a BlueGreen Intelligence Map(BIM)to display cyanobacterial biomass
over the entire surface area of Lake Mattamuskeet.The probe device is a minimally invasive and measures six key water variables(air
temperature,water temperature,turbidity,chlorophyll-a,phycocyanin,ambient light),GPS coordinates,and reports data to the science teams.
16) Purpose/hypothesis:
1)Using historical analysis to construct a precision-based treatment at select treatment areas of Lake Mattamuskeet.Selection of areas will be based on a unanimous decision
between the Responsible Parties.2)Calibrating Lake Guarde View to the unique setup of Lake Mattamuskeet in combination with in-water probe devices to nurture and
enhance Al capabilities in pursuit of improved remote monitoring and studying predictive modelling.3)Collecting data to better understand the phytoplankton behavior within
the entire Lake Mattamuskeet for comparative research-oriented analysis before,during and after Phase 2 Cyanobacterial Algal Bloom Treatment(see Section Phase 2 of
attached proposal).
17) Expected benefits of research/monitoring:
A coropeaherrewe analyst,of the desgnated area with the BlueGreen Intelligence Map?BIM)by.Collecting and analysing remote semen satellite imagery and avai0Me reslon,.al data surveying all inflow and ou!&,o of designated area waterways.autcwrerng for
c onthb,ltng hydrological aspects to understand pa dynamics of water flow and creme movements'.and modeling phyaut banners water movement.semo-erttJneed areas.shalow water areas.sewage systems Moan.near designated area,,and the nature at their
discharges.mines in the surety,area stakeholders'input geological features.macro-e0dog1cel ca,e deratbns.where applicable.2j A detennoalon of be most likely areas prone to cyanobacienel infettafon,source of cyanobectenal contaminator,scale of affected
area and eta intensify of the cyaebe tonal[Mums 7)sitespeofc data is integrated into are Al system wet estersove BkeGreen parameters to determine.
I When to treat the area
2.sow much BlueGreen product is needed to etirtrnate the toxic algal Monroe
2.Velma b strategically apply the product.
4 How ogen to repeat treatment to prevent recurrence
18) Briefly describe project history and context of research/monitoring project:
The North Carolina Collaboratory(Collaboratory)requested proposals from vendors capable of implementing an In-situ treatment of nutrient impaired surface waters to remediate and prevent
cyanobacterial harmful algal blooms in lakes and reservoirs of North Carolina.BlueGreen Water Technologies(herein referred to as BlueGreen)was awarded in July 2022 a contract by execution of
a University of North Carolina purchase order pursuant to BlueGreen's IFB response for the study and treatment of Harmful Algal Blooms(HABs)In select water bodies within North Carolina.
8lueGreen Water Technologies has invested years in populating a vast database—a distinct repository containing thousands of global BlueGreen assessments.Integrating this ever-growing
database with the constantly evolving artificial intelligence(All models.BlueGreen leverages data-based science to successfully mitigate algal blooms.
19) Briefly describe project's relationship to other research/monitoring projects either known of or conducted by the applicant:
N/A.
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FWS Form 3-1383-R(Rev. 03/2020) OMB Control No 1018-0102
U.S Department of the Interior Expires 01/31/2024
20)Identify the types of specimen collections to be taken(see specimen collection clause in the instruction section#20)or data to be collected
during the proposed project:
Theprobe device is minimallyinvasive and measures six keywater variables(air temperature,water temperature,
P P
turbidity, chlorophyll-a, phycocyanin, ambient light), GPS coordinates, and reports data to the science teams. It is solar
powered, cellular connected, and has selfcleaning sensors that collect data 24/7, updating every 30 minutes.
21) List other cooperators and institutions involved in the project
The North Carolina Collaboratory and the University of North Carolina at Chapel Hill.
22)Generally identify the anticipated timeline for analysis,write-up and publication:
Deployment December 2022
Monitoring:December 2022-December 2024
Analysis:Continuous for the duration of deployment
Write-up.As needed for the duration of deployment
Publication:Final Report upon completion of project December 2024
23) For research involving animals,attach an Assurance of Animal Care Form or an approval from an Institutional Animal Care and Use
Committee? Is a form or approval attached? O Yes O No O N/A
License/Insurance/Certifications/Permits
Note: Contact the specific refuge office where the research project is going to be conducted to determine if any type of license,
insurance, certification(s),or permit(s)will be required.We may process this Special Use Permit while the applicant obtains them.
Attach additional sheets to the application if the text spaces provided are inadequate.
24a)List and attach copies of any licenses you have for equipment operation(i.e., aviation or commercial boats), pesticide applications,
transporters)or others if required:
License Type Number Expiration Date (if applicable)
24b)List and attach copies of any insurance you have(i.e general liability,flight/grounding,contaminants.medical evacuation,or others if required:
Insurance Type Carrier Expiration Date (if applicable)
24c)List and attach copies of any certifications you have, such as rat free, hull inspections,CPR/First Aid, or others if required:
Certificate Type Expiration Date(if applicable)
24d)List and attach copies of any other Federal, State,or Tribal permits if required:
Permit Type Permit Number Expiration Date (if applicable)
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Logistics and Transportation Attach additional sheets if the text spacesprovided are inadequate.
9 p
p q
25a) Does project require personnel to stay overnight on the refuge? Yes O No 0
25b) If yes, how many? And list known personnel involved in overnight stay below:
List Names List Names List Names List Names
26)Specifically describe all major instrumentation/equipment/gear(i.e. use of drones)and materials used, if applicable or required:
30-50 floating autonomous water probe devices.
27a)Provide details and schedule for the installation of instrumentation:
These devices will be deployed in December 2022.
27b)Provide details and schedule for the removal of instrumentation:
The duration of deployment will range 12-24 months, dependent on quality of data collected and agreement among the Mattamuskeet
National Wildlife Refuge,the UNC and site stakeholders. Upon completion of the project or at the discretion of both BlueGreen Water
Technologies and Mattamuskeet National Wildlife Refuge,all floating probe devices will be removed, leaving no trace of the device.
27c) If instrumentation is permanent, describe need:
N/A
27d) If instrumentation requires a maintenance schedule,describe needs and schedule:
The devices will be visually checked and cleaned for any biofouling every two months or as needed(as determined by the device's internal maintenance alert warning system).
27e)Provide a data collection schedule:
These devices will automatically collect data 24/7, updating every 30 minutes.
28)Provide logistical arrangements for offsite transportation of samples:
The devices use cellular signal to upload all data to the Internet automatically.
29a)Provide detailed information on the logistics for onsite, intersite,and/or ship-to-shore transportation to or on the refuge,if required:
BlueGreen Water Technologies is responsible for installation and maintenance of in-situ probe devices.
BlueGreen will inform and consult with the Mattamuskeet National Wildlife Refuge on dates of
maintenance as well as vehicle information.
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FWS Form 3-1383-R(Rev. 03/2020) OMB Control No. 1018-0102
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29b)Provide descriptions. license plate and/or identification numbers of vehicles used for onsite transportation, if required:
Vehicle Type Plate/I.D./Registration# Vehicle Type Plate/I.D./Registration#
29c) Provide descriptions, license plate and/or identification numbers of vehicles used for intersite transportation, if required:
Vehicle Type Plate/I.D./Registration# Vehicle Type Plate/I.D./Registration#
29d)Provide descriptions, license plate and/or identification numbers of vehicles used for ship-to-shore transportation, if required
Vehicle Type Plate/I.D./Registration# Vehicle Type Plate/I.D./Registration#
30a) Is fuel cache needed? Yes O No O N/A 0 30b) Provide specific location(s)of fuel caches:(GPS coordinates preferred)
31)Attach Safety Plan if required. Is Safety Plan attached?Yes O No O N/A O
Work and Living Accommodations
32) Specifically describe onsite work and/or living accommodations, including spike camps:
N/A
33) Specifically describe on or offsite hazardous material storage or other on or offsite material storage space: (Including on and offsite fuel caches)
N/A
Sign,date,and print this form and return it to the refuge for processing.By signing this application,I agree my operations will
conform to the information I have provided in this application,and I understand that any deviations or changes to this information
must receive prior written approval.
34) Signature of Applicant: 9J)l�.r,.._ Date of Application:
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FWS Form 3-1383-R(Rev.03/2020) OMB Control No 1018-0102
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NOTICES
No Members of Congress or Resident Commissioner shall participate in any part of this contract or to any benefit that may arise from it, but this
provision shall not pertain to this contract if made with a corporation for its general benefit.
The Permittee agrees to be bound by the equal opportunity"nondiscrimination in employment"clause of Executive Order 11246.
PRIVACY ACT STATEMENT
Authority: The information requested is authorized by the National Wildlife Refuge System Administration Act (16 U.S.C. 668dd-ee)and the
Refuge Recreation Act(16 U.S.C.460k-460k-4).
Purpose: To collect the applicant's information to process permits allowing for: research and monitoring activities by students, universities.or
other non-FWS organizations.
Routine Uses: The information will be used by the refuge's administrative office for processing Research and Monitoring Special Use permits.
More information about the routine uses maybe found in the Systems of Records Notice, FWS-5 National Wildlife Refuge Special Use Permits.
Disclosure: Providing the information is voluntary. However, submission of information is required to process and approve research and
monitoring activity usage on the National Wildlife Refuge System.
PAPERWORK REDUCTION ACT STATEMENT
We are collecting this information subject to the Paperwork Reduction Act(44 U.S.C. 3501)to evaluate the qualifications. determine eligibility,
and document permit applicants and to respond to requests made under the Freedom of Information Act and the Privacy Act of 1974. The
information that you provide is required to obtain or retain a benefit: however,failure to provide all required information is sufficient cause for the
U.S. Fish and Wildlife Service to deny a permit. False, fictitious, or fraudulent statements or representations made in the application may be
grounds for revocation of the Special Use Permit and may be punishable by fine or imprisonment(18 U.S.C. 1001). According to the
Paperwork Reduction Act of 1995,an agency may not conduct or sponsor and a person is not required to respond to a collection of
information unless it displays a currently valid OMB control number. OMB has approved this information collection and assigned control
number 1018-0102.
ESTIMATED BURDEN STATEMENT
The public reporting burden for this information collection varies based on the requested specific refuge use. We estimate the relevant public
reporting burden for the Commercial Activity Special Use Permit Application form(Research and Monitoring)to average 5 hours per response,
including the time for reviewing instructions, searching existing data sources, gathering and maintaining the data needed, and completing and
reviewing the collection of information. Send comments regarding this burden estimate or any other aspect of this collection of information,
including suggestions for reducing this burden,to the Service Information Clearance Officer, U.S. Fish and Wildlife Service, 5275 Leesburg
Pike, MS: PRB(JAO/3W), Falls Church,VA 22041-3803,or via email to Info Coll fws.gov. Please do not send your completed form to this
address.
GENERAL CONDITIONS AND REQUIREMENTS
1) Responsibility of Permittee: We shall consider the pemnittee,by operating on the premises, to have accepted these premises with all
facilities,fixtures,or improvements in their existing condition as of the date of this permit.At the end of the period specified or upon earlier
termination,the permittee shall give up the premises in as good order and condition as when received except for reasonable wear,tear,or
damage occurring without fault or negligence. The permittee will fully repay the Service for any and all damage directly or indirectly resulting
from negligence or failure on his/her part, and/or the part of anyone of his/her associates, to use reasonable care.
2) Operating Rules and Laws: The permittee shall keep the premises in a neat and orderly condition at all times, and shall comply with all
municipal, county, and State laws applicable to the operations under the permit as well as all Federal laws, rules, and regulations governing
national wildlife refuges and the area described in this permit. The permittee shall comply with all instructions applicable to this permit issued
by the refuge official in charge. The permittee shall take all reasonable precautions to prevent the escape of fires and to suppress fires and
shall render all reasonable assistance in the suppression of refuge fires.
3) Use Limitations: The permittee's use of the described premises is limited to the purposes herein specified and does not, unless provided for
in this permit, allow him/her to restrict other authorized entry onto his/her area; and allows the U.S. Fish and Wildlife Service to carry on
whatever activities are necessary for: (1) protection and maintenance of the premises and adjacent lands administered by the U.S. Fish and
Wildlife Service; and (2)the management of wildlife and fish using the premises and other U.S. Fish and Wildlife Service lands.
4)Transfer of Privileges: This permit is not transferable, and no privileges herein mentioned may be sublet or made available to any person or
interest not mentioned in this permit. No interest hereunder may accrue through lien or be transferred to a third party without the approval of the
Regional Director of the U.S. Fish and Wildlife Service and the permit shall not be used for speculative purposes.
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5) Compliance: The U.S. Fish and Wildlife Service's failure to require strict compliance with any of this permit's terms, conditions, and
requirements shall not constitute a waiver or be considered as a giving up of the U.S. Fish and Wildlife Service's right to thereafter enforce any
of the permit's terms or conditions.
6) Conditions of Permit not Fulfilled: If the permittee fails to fulfill any of the conditions and requirements set forth herein,the U.S.Fish and Wildlife
Service shall retain all money paid under this permit to be used to satisfy as much of the permittee's obligation as possible.
7) Payments:All payment shall be made on or before the due date to the local representative of the U.S. Fish and Wildlife Service by a postal
money order or check made payable to the U.S. Fish and Wildlife Service.
8)Termination Policy: At the termination of this permit the permittee shall immediately give up possession to the U.S. Fish and Wildlife Service
representative, reserving, however, the rights specified in paragraph 11 below. If he/she fails to do so, he/she will pay the U.S. Fish and
Wildlife Service, as liquidated damages, an amount double the rate specified in this permit for the entire time possession is withheld. Upon
yielding possession, we will still allow the permittee to reenter as needed to remove his/her property as stated in paragraph 11 below.The
acceptance of any fee for the liquidated damages or any other act of administration relating to the continued tenancy is not to be considered as
an affirmation of the permittee's action nor shall it operate as a waiver of the U.S. Fish and Wildlife Service's right to terminate or cancel the
permit for the breach of any specified condition or requirement.
9) Revocation Policy: The Regional Director of the U.S. Fish and Wildlife Service may revoke this permit without notice for noncompliance with
the terms hereof, or for violation of general and/or specific laws or regulations governing national wildlife refuges, or for nonuse. It is at all times
subject to discretionary revocation by the Director of the Service. Upon such revocation the U.S. Fish and Wildlife Service, by and through any
authorized representative, may take possession of said premises for its own and sole use, and/or may enter and possess the premises as the
agent of the permittee and for his/her account.
10) Damages:The U.S. Fish and Wildlife Service shall not be responsible for: any loss or damage to property including but not limited to crops,
animals, and machinery; injury to the permittee or his/her relatives or to the officers, agents, employees, or any other(s)who are instructed to
be on the premises; the sufferance from wildlife or employees or representatives of the U.S. Fish and Wildlife Service carrying out their official
responsibilities. The permittee agrees to hold the U.S. Fish and Wildlife Service harmless from any and all claims for damages or losses that
may arise to be incident to the flooding of the premises resulting from any associated Government river and harbor, flood control, reclamation,
or Tennessee Valley Authority activity.
11) Removal of Permittee's Property: Upon the expiration or termination of this permit,if all rental charges and/or damage claims due to the U.S.
Fish and Wildlife Service have been paid,the permittee may.within a reasonable period as stated in the permit or as determined by the U.S. Fish
and Wildlife Service official in charge,but not to exceed 60 days, remove all structures, machinery, and/or equipment, etc., from the premises
for which he/she is responsible. Within this period the permittee also must remove any other of his/her property including his/her acknowledged
share of products or crops grown,cut, harvested, stored,or stacked on the premises. Upon failure to remove any of the above items within the
aforesaid period,they shall become the property of the U.S. Fish and Wildlife Service. .
INSTRUTIONS FOR COMPLETING APPLICATION
You may complete the application portion verbally, in person, or electronically and submit to the refuge for review. Note: Please read instructions
carefully as not all information is required for each activity. Contact the specific refuge where the activity will take place if you have questions
regarding the applicability of a particular item. We may add special conditions or permit stipulations to permit prior to approval.
la-1c) Identify if permit application is for new, renewal, or modification of an existing permit, whether or not you have or will be applying to
another refuge for the same activity, and for which refuge(s). Permit renewals may not need all information requested. Contact the specific
refuge headquarters office where the activity is going to be conducted to determine applicability of this requirement.
2-3) Provide principal investigator or applicant full name. Attach principal investigator's Curriculum Vitae or Resume, if required. Permit
renewals generally do not require a Curriculum Vitae or Resume if the project is a continuation of a previously issued permit being
conducted by the same investigator. Contact the specific refuge office to determine applicability of this requirement.
4-9) Provide investigator's physical and/or mailing address, phone, fax, e-mail, affiliation and/or sponsoring organization, and
relationship to affiliation or organization (title, professor, student. etc.).
10) Provide the names and addresses of assistants, subcontractors, or subpermittees. Vla may requie names and addresses if the assistants,
subcontractors or subpermittees will be operating on the refuge without the permittee being present. Volunteers. assistants, subcontractors. or
subpermittees accompanied by the permittee need not be identified.
11) Provide title of research or monitoring project.
12a-12b) Attach a full research or monitoring proposal, if required. Permit renewals generally do not require a project proposal if the project is a
continuation of a previously issued permit being conducted by the same investigator.Contact the specific refuge office to determine applicability
of this requirement.
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13) Provide detailed information on the activity, including timing, frequency, how the project is expected to proceed, etc. Permit renewals may not
need activity description, if the activity is unchanged from previous permit. Most repetitive research projects do not require an activity description
for each visit to the refuge. Contact the specific refuge office to determine applicability of this requirement.
14) Identify specific location(GPS coordinates preferred) if not a named facility,and/orattach a map ofthe location. Permit renewals may not require
a location if the project is essentially unchanged from the previous permit. Contact the specific refuge office to determine applicability of this
requirement.
15)Identify species or habitats being studied.
16-17) Specifically identify purpose or hypothesis of the research or monitoring project and describe expected benefits. Permit renewals may
not need to identify purpose or hypothesis if the project is a continuation of a previously issued permit being conducted by the same
investigator. Contact the specific refuge office to determine applicability of this requirement.
18) Briefly describe project history and context. Permit renewals should describe previous research activities as part of a previously issued
permit being conducted by the same investigator.Contact the specific refuge office to determine applicability of this requirement.
19) Briefly describe project's relationship to other research/monitoring projects either known of or conducted by the applicant, if applicable.
Include a brief statement of how the research or monitoring permit being applied for will add to or supplement other ongoing research or
monitoring on the same,or related, species or habitats. Contact the specific refuge office to determine applicability of this requirement.
20) Identify specimen collections to be taken or types of data to be collected. You may use specimens collected under this permit, any
components of any specimens (including natural organisms, enzymes, genetic materials or seeds), and research results derived from collected
specimens for scientific or educational purposes only. and not for commercial purposes unless you have entered into a Cooperative Research and
Development Agreement (CRADA) with us. We prohibit the sale of collected research specimens or transfers to third parties for commercial
purposes. Breach of any of the terms of this permit will be grounds for revocation of this permit and denial
of future permits. Furtheore. if you
sell or otherwise transfer for commercial purposes collected specimens, arty components thereof, or any products or research results developed
from such specimens or their components without a CRADA, you will pay us a royalty rate of 20 percent of gross revenue from such sales. In
addition to such royalty,we may seek other damages and injunctive relief against you. Permit renewals may not need to identify samples taken if
the project is a continuation of a previously issued permit being conducted by the same investigator. Contact the specific refuge office to
determine applicability of this requirement.
21) List other cooperators and institutions involved in the project, if applicable. Contact the specific refuge office to determine applicability of this
requirement.
22)Generally, identify the anticipated time line for analysis,write-up, and publication of project results. Include whether the project is a single, or
multiple year project. Identification of an actual publication where the results are printed is not necessary. However, applicants should include
the anticipated dissemination of project results. Contact the specific refuge office to determine applicability of this requirement.
23) Check box acknowledging a completed Assurance of Animal Care Form or an Institutional Animal Care and Use Committee (or
equivalent) that has granted approval has been completed,and has been submitted to refuge station,if required.Contact the specific refuge office
to determine applicability of this requirement.
24a-24d) Specifically identify types and numbers of licenses, insurance, certifications, and other State, Federal, or Tribal permits if required.
Contact the specific refuge headquarters office where the project is going to be conducted to determine applicability of these requirements,
and to coordinate the simultaneous applications of any of these requirements while this Special Use Permit is being processed.
25a-25b) Provide the number of and/or name(s)of any personnel required to stay overnight on the refuge, if applicable.
26) Identify all equipment(including drones) and materials that will be used, if required. Permit renewals may not require a list of equipment if
the project is essentially unchanged from a previously issued permit. Contact the specific refuge office to determine applicability of this
requirement.
27a-27e) Identify types and schedule(s) of installation of any instrumentation, data collection, and maintenance schedule of instrumentation, if
required. Permit renewals may not require a list of equipment if the project is essentially unchanged from a previously issued permit. However,
schedules of installation of any instrumentation, data collection, and maintenance schedule of instrumentation may still be required. Contact the
specific refuge headquarters office where the project is going to be conducted to determine applicability of this requirement.
28)Identify logistical arrangements for offsite transportation of samples taken, if applicable.
29a-29d) Describe and provide vehicle descriptions and license plate or identification numbers of all vehicles, including boats and airplanes, if
required. Motor vehicle descriptions are only required for permittee vehicle, and/or if the vehicle will be operated on the refuge without the
permittee being present. Motor vehicles that are accompanied by the permittee as part of a group (convoy) activity need not be identified if
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cleared in advance by refuge supervisor. Specifically describe ship-to-shore, intersite (between islands, camps, or other sites) and onsite
transportation mechanisms, and license plate or identification numbers, if required.
30a-30b) Identify specific location(s)of fuel cache(s) (GPS coordinates preferred), if required.
31a-31b)Attach safety plan, if required. Contact the specific refuge office to determine applicability of this requirement.
32) Specifically describe onsite work and/or living accommodations, if required. Include descriptions and locations (GPS coordinates preferred)
of spike camps or other remote work and/or living accommodations that are not part of the base of operations. Contact the specific refuge
office to determine applicability of this requirement.
33) Specifically describe onsite and offsite hazardous material storage, or other onsite material storage space (including on and offsite fuel
caches), if required. Contact the specific refuge office to determine if descriptions of hazardous material storage or other onsite material
storage are required.
34) Sign, date, and print the application. Click on the Print button to print the application (if using the fillable version). The refuge official will
review and, if approved,fill out the remaining information, sign, and return a copy to you for signature and acceptance.
THIS APPLICATION FORM IS NOT VALID AS A PERMIT
BUT MAY BE USED AS A REFERENCE DOCUMENT ATTACHED TO THE OFFICIAL PERMIT.
ONLY OFFICIAL REFUGE PERSONNEL MAY ASSIGN A VALID PERMIT NUMBER AND PERMIT TERM
TO THIS APPLICATION FORM AFTER THE PERMIT HAS BEEN APPROVED.
Page 9 of 9

6 BlueGreen DATE: TO:
November 22,2022 Kendall Smith
WATER TECHNOLOGIES
PHASE 1
RESEARCH TOPIC/TITLE: In-Situ Water Quality Monitoring
T PROJECT DESCRIPTION N
BACKGROUND
The North Carolina Collaboratory (Collaboratory)requested proposals from vendors capable
of implementing an in-situ treatment of nutrient impaired surface waters to remediate and
prevent cyanobacterial harmful algal blooms in lakes and reservoirs of North Carolina. This
Invitation for Bids (IFB)was solicited on behalf of the North Carolina General Assembly, as
provided in Section 8.18 of Session Law 2021-180, as amended by Section 2.17 of Session
Law 2022-6. BlueGreen Water Technologies (herein referred to as BlueGreen)was awarded
July 2022 a contract by execution of a University of North Carolina purchase order pursuant
to BlueGreen's IFB response for the study and treatment of Harmful Algal Blooms (HABs) in
select water bodies within North Carolina. Contract Reference: IFB3000010521.
BlueGreen Customer/Partner: University of North Carolina (UNC) at Chapel Hill
AIMS
• Using historical analysis to construct a precision-based treatment at select treatment
areas of Lake Mattamuskeet. Selection of areas will be based on a unanimous
decision between the Responsible Parties.
• Calibrating Lake Guard®View to the unique setup of Lake Mattamuskeet in
combination with in-water probe devices to nurture and enhance Al capabilities in
pursuit of improved remote monitoring and studying predictive modelling.
• Collecting data to better understand the phytoplankton behavior within the entire
Lake Mattamuskeet for comparative research-oriented analysis before,during and
after Phase 2 Cyanobacterial Algal Bloom Treatment(see Section Phase 2).
RESPONSIBLE PARTIES
BlueGreen Water Technologies is responsible for installation and maintenance of in-situ
probe devices. BlueGreen will inform and consult with the Mattamuskeet National Wildlife
Refuge, the UNC and site stakeholders throughout project progression. BlueGreen will share
raw data from probe devices with the Mattamuskeet National Wildlife Refuge, the UNC and
site stakeholders upon request.
METHODOLOGY
BlueGreen Water Technologies has invested years in populating a vast database —a distinct
repository containing thousands of global BlueGreen assessments. Integrating this ever-
CONTACT: .11r.
) OO
.%
1-866-269-2828 [e-mail] 301 South Hills Village Ste
www.bluegreenwatertech.com LL200#452,Pittsburgh,PA
15241

1
1

6 BlueGreen DATE: TO:
November 22,2022 Kendall Smith
WATER TECHNOLOGIES
growing database with the constantly evolving artificial intelligence (Al) models, BlueGreen
leverages data-based science to successfully mitigate algal blooms. To that end, 30-50
floating autonomous water probe devices will be temporarily deployed at strategic locations
across Lake Mattamuskeet. BlueGreen will consult and come to an agreement with
Mattamuskeet National Wildlife Refuge on placement of probe devices. The probe device is
a minimally invasive and measures six key water variables (air temperature, water
temperature,turbidity, chlorophyll-a, phycocyanin, ambient light), GPS coordinates, and
reports data to the science teams. It is solar powered, cellular connected, and has self-
cleaning sensors that collect data 24/7, updating every 30 minutes. Each probe device will
be visually checked and cleaned for any biofouling every two months or as needed per the
device's internal maintenance alert warning system. The duration of deployment will range
12-24 months, dependent on quality of data collected and agreement among the
Mattamuskeet National Wildlife Refuge, the UNC and site stakeholders. Upon completion of
the project or at the discretion of both BlueGreen Water Technologies and Mattamuskeet
National Wildlife Refuge, all floating probe devices will be removed, leaving no trace of the
device.
Data collected from all probe devices will contribute to the creation of a BlueGreen
Intelligence Map (BIM)to display cyanobacterial biomass over the surface area. The
methodology uses remote imaging-sourced waterbody evaluation capabilities. The
BlueGreen data science team and local field experts will collaborate to assess the specific
water problem. The data scientists will source up to three years of historic and current
satellite image data for the area in question. From these data, a BlueGreen Intelligence Map
(BIM) of the waterbody that details both the past and present algal bloom status will be
created, including depicting spread over the water surface and at depth in the water column.
Ultimately,this 3-dimensional time-sensitive map displays cyanobacterial biomass on the
surface area by time of year.
Expected outcomes, significance or rationale
• Comprehensive analysis of designated area with the BlueGreen Intelligence Map
(BIM).
• Collect and analyze remote sensing satellite imagery and available historical
data.
o Survey all inflow and outflow of designated area waterways.
u Account for contributing hydrological aspects to understand the dynamics of
CONTACT:
1-866-269-2828 [e-mail] 301 South Hills Village Ste
www.bluegreenwatertech.com LL200#452,Pittsburgh,PA
15241
1

l _-

6 BlueGreen DATE: TO:
November 22,2022 Kendall Smith
WATER TECHNOLOGIES
water flow and current movements.
o Model physical barriers,water movement, semi-enclosed areas, shallow
water areas, sewage systems, industries near designated areas and the
nature of their discharges, mines in the vicinity, area stakeholders' input,
geological features, macro-ecological considerations, where applicable.
• Determining the most likely areas prone to cyanobacterial infestation, source of
cyanobacterial contamination, scale of affected area, and the intensity of the
cyanobacterial blooms.
• Site-specific data is integrated into the Al system with extensive BlueGreen
parameters to determine:
1.When to treat the area.
2. How much BlueGreen product is needed to eliminate the toxic algal blooms.
3.Where to strategically apply the product.
4. How often to repeat treatment to prevent recurrence.
Timetable
Deployment: December 2022
Monitoring: December 2022-December 2024
Analysis: Continuous for the duration of deployment
Write-up:As needed for the duration of deployment
Publication: Final Report upon completion of project December 2024
CONTACT: ';;' 00
.v.
1-866-269-2828 [e-mail] 301 South Hills Village Ste
www.bluegreenwatertech.com LL200#452,Pittsburgh,PA
15241

ttA STq
41 -� UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
0 F WASHINGTON, D.C. 20460
0
~r4L PROtE��
OFFICE OF CHEMICAL SAFETY
AND POLLUTION PREVENTION
March 6, 2023
Brian Hogan
Agent for Bluegreen US Water Technologies, Inc.
do KRK Consulting LLC
5807 Churchill Way
Medina,0 44256
Subject: Labeling Notification per Pesticide Registration Notice(PRN)98-10—Delete statement
related to shallow water
Product Name: Lake Guard Oxy
EPA Registration Number: 93647-2
Application Date: 01/30/2023
OPP Case Number: 00428733
Dear Mr. Hogan:
The U.S. Environmental Protection Agency (EPA)is in receipt of your application for notification under
Pesticide Registration Notice(PRN)98-10 for the above referenced product. The Biopesticides and
Pollution Prevention Division(BPPD)has conducted a review of this request for its applicability under
PRN 98-10 and finds that the action requested falls within the scope of PRN 98-10.
The labeling submitted with this application has been stamped"Notification"and will be placed in our
records. You must submit one(1)copy of the final printed labeling with the modifications.
At this time,the U.S. Environmental Protection Agency (EPA)is not requiring a change to the
restricted-entry interval (REI)of 0 hours(for non-spraying methods)and 1 hour(for spraying methods)
as it appears on your labeling in the Agricultural Use Requirements Box. The EPA is currently
developing guidance and criteria for when it will allow a REI of less than 4 hours and might require a
change to the REI for this product in the future.
Should you wish to add/retain a reference to your company's website on your label,then please be
aware that the website becomes labeling under the Federal Insecticide, Fungicide,and Rodenticide Act
(FIFRA)and is subject to review by the EPA. If the website is false or misleading,the product will be
considered to be misbranded and sale or distribution of the product is unlawful under FIFRA section
I2(a)(I)(E). 40 CFR § 156.10(a)(5) lists examples of statements the EPA may consider false or
misleading. In addition,regardless of whether a website is referenced on your product's label,claims
made on the website may not substantially differ from those claims approved through the registration
process. Therefore, should the EPA find or if it is brought to our attention that a website contains false

Page 2 of 2
EPA Reg. No. 93647-2
OPP Case No. 00428733
or misleading statements or claims substantially differing from the EPA-approved registration,the
website will be referred to the EPA's Office of Enforcement and Compliance Assurance.
If you have any questions,please contact Susannah Powell via email at powell.susannahepa.gov.
Sincerely,
James Parker, Team Leader
Biochemical Pesticides Branch
Biopesticides and Pollution
Prevention Division (75 l 1 P)
Office of Pesticide Programs

NOTIFICATION
93847-2
03/0e/2023 LAKE GUARD® Oxy
Algaecide/Cyanobacteriocide
[Large Granules] [Small Granules] [Dust]
ACTIVE INGREDIENT
Sodium percarbonate 83.3%
OTHER INGREDIENTS . 16.7%
TOTAL 100.0% Certified to
NSF/ANSIiCAN 60
[UN 3378} MUL 33 mg/L
[CLASS 5.1]
KEEP OUT OF REACH OF CHILDREN
DANGER/ PELIGRO
Si usted no entiende la etiqueta, busque a alguien para que se la explique a usted en detalle.
(If you do not understand the label,find someone to explain it to you in detail.)
FIRST AID
If in eyes • Hold eye open and rinse slowly and gently with water for 15-20 minutes. Remove
contact lenses, if present, after the first 5 minutes, then continue rinsing.
• Call a poison control center or doctor for treatment advice.
If swallowed • Call a poison control center or doctor immediately for treatment advice.
• Have person sip a glass of water if able to swallow.
• Do not induce vomiting unless told to by a poison control center or doctor.
• Do not give anything to an unconscious person.
If on skin • Take off contaminated clothing.
• Rinse skin immediately with plenty of water for 15-20 minutes.
• Call a poison control center or doctor for treatment advice.
If inhaled • Move person to fresh air.
• If person is not breathing, call 911 or an ambulance, then give artificial respiration,
preferably mouth-to-mouth if possible.
• Call a poison control center or doctor for further treatment advice.
Have the product container or label with you when calling a poison control center, doctor, or going for
treatment. For non-emergency information concerning this product, call the National Pesticides
Information Center(NPIC) at 1-800-858-7378(NPIC Web site:www.npic.orst.edu). For emergencies,
call the poison control center 1-800-222-1222.
NOTE TO PHYSICIAN: Probable mucosal damage may contraindicate the use of gastric lavage.
EPA Reg. No. 93647-2
EPA Est. No.XXXXX-XX-X
Net Contents: 2, 5, 10,20, 25, 50, 500, 1,000, 2,000, 2,204 lbs.
Batch Code:
Page 1 of 5

PRECAUTIONARY STATEMENTS
Hazards to Humans and Domestic Animals
DANGER/PELIGRO
Corrosive. Causes irreversible eye damage and causes skin burns. May be fatal if swallowed. Harmful if
absorbed through skin or inhaled. Do not get in eyes, on skin, or on clothing. Avoid breathing dust. Wear
protective eyewear, such as goggles, face shield, or safety glasses. Wash thoroughly with soap and
water after handling and before eating, drinking, chewing gum, using tobacco or using the toilet. Remove
and wash contaminated clothing before reuse.
Environmental Hazards
This pesticide is toxic to birds. Do not apply this product or allow it to drift to blooming crops or weeds
while pollinating insects are actively visiting the area.
For container sizes 50 lbs. or greater: Do not discharge effluent containing this product into lakes,
streams, ponds, estuaries, oceans or other waters unless in accordance with the requirements of a
National Pollutant Discharge Elimination System (NPDES) permit and the permitting authority has been
notified in writing prior to discharge. Do not discharge effluent containing this product into sewer systems
without previously notifying the local sewage treatment plant authority. For guidance contact your State
refer to the product
Water Board or Regional Office of U.S. EPA. For additional information,
Material Safety Data Sheet.
Physical and Chemical Hazards
Oxidizing agent. Never use with other pesticides, cleaners, or oxidizing agents.
Personal Protective Equipment
Corrosive. Mixers, loaders. applicators, and other handlers must wear the following:
• Long-sleeved shirt and long pants
• Chemical resistant gloves
• Protective eyewear(googles or face shield)
• Shoes plus socks
Follow manufacturer's instructions for cleaning/maintaining PPE. If no such instructions for washables
exist, use detergent and hot water. Keep and wash PPE separately from other laundry.
USER SAFETY RECOMMENDATIONS
Users should remove clothing/PPE immediately if pesticide gets inside. Then wash thoroughly and put
on clean clothing.
Users should remove PPE immediately after handling this product. Wash the outside of gloves before
removing. As soon as possible, wash thoroughly and change into clean clothing.
DIRECTIONS FOR USE
It is a violation of Federal Law to use this product in a manner inconsistent with its labeling.
GENERAL APPLICATION RESTRICTIONS
Do not apply this product in a way that will contact workers or other persons. either directly or through
drift. Only protected handlers may be in the area during application. For any requirements specific to your
State or Tribe, consult the State or Tribal agency responsible for pesticide regulation.
This product is not intended as treatment against any public health organism for any use on this label.
Do not apply directly to treated,finished drinking water reservoirs or drinking water receptacles when the
water is intended for human consumption.
SPRAY DRIFT MANAGEMENT
Page 2 of 5

A variety of factors including weather conditions (e.g., wind direction, wind speed, temperature, and
relative humidity)and the method of application(e.g., ground or aerial)can influence pesticide drift. The
applicator must evaluate all factors and make appropriate adjustments when applying this product.
Wind Speed
Do not apply at wind speeds greater than 15 mph. Only apply this product if the wind direction favors on-
target deposition(approximately 3 to 10 mph), and there are no sensitive areas within 250 feet downwind.
Temperature Inversions
If applying at wind speeds less than 3 mph,the applicator must determine if a)conditions of temperature
inversion exist, or b) stable atmospheric conditions exist at or below nozzle height. Do not make
applications into areas of temperature inversions or stable atmospheric conditions.
Other State and Local Requirements
Applicators must follow all state and local pesticide drift requirements regarding application of hydrogen
peroxide compounds. Where states have more stringent regulations, they must be observed.
Equipment
All aerial and ground application equipment must be properly maintained and calibrated using appropriate
carriers or surrogates.
For aerial applications:
The boom length must not exceed 75% of the wingspan or 90% of the rotor blade diameter. Release
granules at the lowest height consistent with efficacy and flight safety. When applications are made with
a crosswind, the swath must be displaced downwind. The applicator must compensate for this
displacement at the up and downwind edge of the application area by adjusting the path of the aircraft
upwind.
GENERAL INFORMATION
Water bodies, such as impounded water, raw-water for drinking, lakes, ponds, reservoirs, waste water,
irrigation, drainage, aquaculture, conveyance ditches, pipes, canals, laterals, estuaries, bayous, lagoons,
and brackish and salt, sea and ocean water contaminated with algae/cyanobacteria can be treated with
Lake Guar&Oxy.
Do not apply when wind speed favors drift beyond the area intended for treatment.
stagnant. Do not use the product to treat ornamental fish(i.e.,fish that grow in home aquariums or
outdoor tanks for aesthetic purposes).
Water hardness, temperature of the water, type and cell density of algae/cyanobacteria to be controlled,
and the amount of water flow are to be considered in using Lake Guard Oxy to control
algae/cyanobacteria. Treated water resource should be monitored systematically for the presence of
harmful algae/cyanobacteria using adequate apparatuses. Begin treatment as soon algal/cyanobacterial
cell numbers reach 5,000-20,000 cells/mL (and below 10 pg chlorophyll-a/liter).
If treatment is delayed until algal/cyanobacterial cell numbers exceed 20,000 cell/mL(or equivalently,
above 10 pg chlorophyll-a/liter), an increase in the quantities of Lake Guar&Oxy will be required, as well
as in treatment frequency. Use caution when treating heavy blooms, as oxygen loss from mass
decomposition of dead algae/cyanobacteria can cause fish suffocation. See application rates section for
treating heaving blooms.
Always apply the granules off wind and let the wind and currents to carry them where
algae/cyanobacteria cell masses are concentrated. It is best to treat algae on a sunny day at morning
hours. If there is some doubt about the amount of Lake Guard'Oxy to apply, it is best to start with the
lower application rate. The quick and easy treatment application(minutes for applying a hundred Ib), and
Page 3 of 5

the visual results that would be apparent within 48-72 hours, enable the applicator to make a quick
adjustment on whether to increase the dose rate by 50-100% during the next treatment application, or
scale it back and use even less product under similar conditions. Since all water bodies are unique
aquatic ecosystems, and, therefore, differ from each other in treatment response and longevity. the Lake
Guard®Oxy application protocol offers the applicator a scalable dose rate that saves money and reduces
the chemical load introduced to the environment. For first time users, or when in doubt, applicators should
start with 0.5-5 lb/acre and adjust the dose rates (increase or decrease) by increments of 25-50%. and
over time can fine-tune the treatment application for each waterbody.
APPLICATION RATES
The best method by which to apply Lake Guard®Oxy granules to water is by broadcasting(dusting) it
over a well-defined contamination zone, at early bloom stages,when harmful algal/cyanobacteria
numbers are at 5,000 to 20,000 cells/mL(and below 10 pg chlorophyll-a/liter).
Determination of surface area to be treated
Determine the size of the infested area as follows: (1) in small infested reservoirs, under than 250 acres,
obtain surface area by measuring of regular shaped ponds or mapping of irregular ponds or by reference
to previously recorded engineering data or maps. (2) In water bodies larger than 250 acres (or smaller
ponds with a defined contaminated zone)outline the infested area by a combination of the following
instruments: microscopical count, pigment extraction, toxin evaluation, probes that detect specific
pigments that are known to serve as a correlated proxy for algae/cyanobacteria biomass, satellite
imaging, etc. NOTE: evaluation of the state of the infestation should be done by professional personnel.
Determination of the application rate
For control of harmful algae/cyanobacteria infestation it is essential to begin Lake Guard®Oxy treatment
when harmful algae/cyanobacteria cell numbers are in the range of 5,000-20,000 cell/mL(or below 10 pg
chlorophyll-a/liter). Apply 0.5-5 lbs./acre Lake Guard®Oxy at these algal/cyanobacterial cell-densities.
Always start with the lower rate. At higher infestation rates,when cyanobacterial cell density is between
20,000-100,000 cells/mL(or between 10-50 pg chlorophyll-a/liter) use 5-301bs./acre Lake Guard®Oxy.
If treatment is delayed until algal/cyanobacterial cell numbers exceed 100,000 cell/mL(or equivalently,
above 50 pg chlorophyll-a/liter), an increase in the quantities of the Lake Guard®Oxy wilt be required, as
well as in treatment frequency.Therefore, in heavy blooms.when cyanobacterial scum or aggregates are
visible to the naked eye (more than 100,000 cells/mL of algae/cyanobacteria or over 50 pg chlorophyll-
a/liter), treat with doses between 30-98 lbs./acre. If doses exceed 98 lbs./acre, treat no more than one-
half of the water area in a single application. Maximum single application rate allowed should not exceed
294 lbs./acre of the Lake Guard®Oxy. NOTE: when cyanobacterial aggregated could be seen with the
naked eye, the cyanobacterial cell density in the water is estimated to exceed 100,000 cells per ml.
When a single application dose is below 30 lbs./acre, minimum retreatment interval is 12 hours.When a
single application dose is between 30-98 lbs./acre, minimum retreatment interval is 24 hours. When a
single application dose exceeds 98 lbs./acre, the minimum retreatment interval is 48 hours.
APPLICATION METHOD
Apply Lake Guard®Oxy using equipment designed for granular dusting. Dusting can be done manually by
hand or by a boat or airplane, depending on the area of the zone and its proximity to reservoir's bank.
When a small duster is mounted on a properly equipped boat, application can be broadcast directly on
the water surface at the edge of the infested zone. Note that the direction of the wind is an important
factor-always dust off-wind. Do not use this method unless completely familiar with this type of
application.
Page 4 of 5

1

STORAGE AND DISPOSAL
Do not contaminate water,food, or feed by storage or disposal.
PESTICIDE STORAGE: Keep pesticide in original container. Do not use in food or drink containers.
PESTICIDE DISPOSAL: Pesticide wastes may be hazardous. Improper disposal of excess pesticide,
spray, mixture or rinsate is a violation of Federal Law. If these wastes cannot be disposed of by use
according to label instructions, contact your State Pesticide or Environmental Control Agency, or the
Hazardous Waste Representative at the nearest EPA Regional Office for guidance.
CONTAINER HANDLING: Nonrefillable container. Do not reuse or refill this container. Completely
empty bag into application equipment, then offer for recycling if available or dispose of empty bag in a
sanitary landfill or by incineration.
CONDITIONS OF SALE AND LIMITATIONS OF WARRANTY AND LIABILITY
Read the entire directions for use, conditions of warranties and limitations of liability before using this
product. If terms are not acceptable, return the unopened product container at once. By using this
product, user or buyer accepts the following Conditions, Disclaimer of Warranties and Limitations of
Liability. CONDITIONS: The directions for use of this product are believed to be adequate and must be
followed carefully. However, it is impossible to eliminate all risks associated with the use of this product.
Ineffectiveness or other unintended consequences may result because of such factors as weather
conditions, presence of other materials, or the manner of use or application, all of which are beyond the
control of BlueGreen Water Technologies. All such risks shall be assumed by the user or buyer.
DISCLAIMER OF WARRANTIES: To the extent consistent with applicable law, BlueGreen Water
Technologies makes no other warranties, express or implied, of merchantability or of fitness for a
particular purpose or otherwise, that extend beyond the statements made on this label. No agent of
BlueGreen Water Technologies is authorized to make any warranties beyond those contained herein or to
modify the warranties contained herein. To the extent consistent with applicable law. BlueGreen Water
Technologies disclaims any liability whatsoever for special, incidental or consequential damages resulting
from the use or handling of this product. LIMITATIONS OF LIABILITY: To the extent consistent with
applicable law, the exclusive remedy of the user or buyer for any and all losses, injuries or damages
resulting from the use or handling of this product, whether in contract, warranty, tort, negligence, strict
liability or otherwise, shall not exceed the purchase price paid or at BlueGreen Water Technologies'
election, the replacement of product.
Property rights are protected under the patent legislation of- USA Patent No. 10,729,138 and USA Patent
No. 10, 092, 005; Indian Patent no. 201617001647; Mexican Patent No. MX/a/2016/000199; South
African Patent No. 2016/00478; the Russian Federation Patent No. 2687929.
Manufactured for:
BlueGreen US Water Technologies, Inc.
Address: 301 South Hills Village Ste LL200#452 Pittsburgh, PA 15241
[e-mail]
[e-mail]
Page 5 of 5

Page 1 of 6
SAFETY DATA SHEET BGT-005
Prepared to OSHA,ACC,ANSI,WHSR.W`Hh11S.20201878/EU Standards I SOS Reoston 2 2 I SOS Revision Date- 3:12 2024
1. PRODUCT 8 COMPANY IDENTIFICATION
II Product Name: BLUEGREEN` ' LAKE GUARD' OXY
1.2 Chemical Name Sodium Percarbonate Mixture
1.3 Sperm,. NA
1 4 T.ade Names. BlueGreena Lake Guards'Oxv,EPA Reg No.93647-2
S Product Use. Algaecide/Biocide
8 Disrbuwrs*lame BlueGreen Water Technologies.Ltd
?s:nbucors Address• 16 HaMiktsoot Blvd,Modi in-Maccabim-Re'ut, Israel 7178096
=merge^ty=ixvie CHEMTEL [phone]/I+1 [phone]
•a Eusness Phdre Fay•E'-.ad: Tel [phone]//E-mail:infoabluegreenwatertech corn
2. HAZARDS IDENTIFICATION
= '3za^''lerVicaDor^ Prepared in accordance with UN Globally Harmonized standards. Intended to comply with OSHA 29 CFR 1910.1200_
Canadian WHMIS and Australian Work Health and Safety.
DANGER! MAY INTENSIFY FIRE - OXIDIZER. MAY BE HARMFUL IF SWALLOWED. CAUSES SERIOUS EYE
DAMAGE.
Classificatioa Ox Sol.2,Acute Tox.coral)5,Eye Dam. 1
- label Eemems Swat Word:DANGER
Hazard Statement%(H) H272-May intensify fire-oxidizer. H303-May be harmful if swallowed.
H318-Causes serious eye damage.
Precautionary Statement;:(P):P210-Keep away from heat,hot surfaces,sparks.open flames,and to .
other ignition sources No smoking P220 - Keep away from clothing and other combustible
materials-P264+P265-Wash hands and exposed skin areas thoroughly with soap and water after
handling. Do not touch eves. P280 - Wear protective gloves/ protective clothing/eye protection/
face protection. P301+P317 - IF SWALLOWED. Get medical help P305+P354+P338 - IF IN Fx
EYES:Immediately rinse with water for several minutes. Remove contact lenses if present and easy .�. �.
to do-continue nnsing.P317-Get medical help. P370+P378-In case of fire Use dry sand,dry
chemical, or alcohol-resistant foam for extinction. P501 - Dispose of contents/container to a
licensed treatment,storage,or disposal facility(TSDF)
2 3 Ot^" `"'a^"b KEEP OUT OF REACH OF CHILDREN.
3. COMPOSITION & INGREDIENT INFORMATION
EXPOSURE LSMITS IN AIR(myms)
ACfi1H NOHSC OSHA
PPm s>vm pvm
ES- ES- ES-
CHEMICAL NAMEIS) CM No. RTECS No EINECS No. % TLV STEL TWA STEL PEAK PEL STEL IDEA OTHER
SODIUM PERCARBONATE 15630-89-4 FG0750000 239-707-6 98 NA NA NE NF NE NA NA NA
Ox Sol 2.Acute Tox.(oral)4,Eye Dam 1 H272,H302,H318
PROPRIETARY INGREDIENTS NA INA INA 12 1 NA 1 NA 1 NF 1 NE I NF I NA I NA I NA
4. FIRST AID MEASURES
4 =rt1 a.i Ingestion: If ingested. do NOT induce vomiting. Rinse mouth with water. Never give anything by mouth to an
unconscious person. Contact the nearest Poison Control Center or local emergency number Provide an
estimate of the time at which the material was ingested and the amount of the substance that was
swallowed
Splashes are not likely, however, if product gets in the eyes. flush with copious amounts of lukewarm
water for at least 15 minutes lifting upper and lower lids,occasionally Get medical attention immediately.
k r: IF ON SKIN: Wash with plenty of soap and water Wash contaminated clothing before reuse.Wash off
immediately with soap and plenty of water for at least 15 minutes.If symptoms persist,call a physician.
inhalation. Remove victim to fresh air at once. If breathing difficult,administer oxygen. If breathing stops give artificial
respiration Keep person warm,quiet and get medical attention.
4 2 E"'is Ingestion: If product is swallowed.may cause nausea.vomiting and/or diarrhea.
)fig Severely irritating to the eyes Symptoms of overexposure may include redness, itching, irritation, and
watering.
Skim May be irritating to skin Symptoms of overexposure may include redness. itching, imtation or burning
sensation
Inhalation: Persons accidentally exposed to dust, particularly during loading and unloading, may experience some
effects including cough wheezing and upper respiratory tract irritation.

1
1
1
I

Page 2of6
SAFETY DATA SHEET BGT-005
Prepared to OSHA.ACC.ANSi,WHSR.WHMIS,2020/878/EU Standards I SOS Revision 2 2 I SDS Revision Date:3112t2024
4. FIRST AID MEASURES — cont'd
4 3 Symptoms of Oyerenoasore Ingestion. Severe irritation.nausea.abdominal pain.vomiting and/or diarrhea
Evers Overexposure in eyes may cause redness.itching.swelling and watering
Symptoms of skin overexposure may include redness,itching.and irritation of affected areas.
Inhalation: Coughing.imtation of mucous membranes and upper respiratory tract
4.4 Acute Health Effects Severe irritation to eyes Additionally,high concentrations of dusts can cause dizziness,headaches,and nausea.
4 5 c^ro^o meaith Effects Overexposure may trigger asthma-like symptoms in some sensitive individuals. May also induce skin sensitization and
respiratory hypersensitivity
4 9 Target Organs: Eyes,Respiratory System
4 7 P.eocai Cond5,ors Pre-existing disorders of the target organs(eyes.lungs). HEALTH 2
apyrr.ated by Exposure: FLAMMABILITY 0
1
PROTECTIVE EOUIPMENT X
EYES I LUNGS I
5. FIREFIGHTING MEASURES
=re&Eiaics+on Hazao: Oxidizing Oxygen released in thermal decomposition may support combustion or accelerate burning
when involved in a fire Contact with combustible material may cause fire Contact with flammables
may cause fire or explosions. May ignite combustibles (wood paper, oil, clothing, etc) Risk of
explosion if heated under confinement. May decompose explosively when heated or involved in a fire.
During a fire, irritating and toxic gases may be generated by thermal decomposition or combustion.
Hazardous combustion products include oxides of carbon(CO.CO2)and oxygen. Runoff may create
fire or explosion hazard
=.u^ear • Metnoas Use water. Do not use dry chemicals or foams. CO; or Halon may provide limited control. Flood fire
area with water from a distance Move containers from fire area if you can do it without nsk Cool iffr\;\
containers with flooding quantities of water until well after fire is out.
•TreigfOang Thor dw!s Firefighters should wear self-contained breathing apparatus and full firefighting turnout gear Use
personal protection equipment. Do not move cargo or vehicle if cargo has been exposed to heat.
oxidizer.May ignite combustibles(wood paper,oil,clothing,etc).Move containers from fire area if you
can do it without nsk. Fight fire from maximum distance or use unmanned hose holders or monitor
nozzles.ALWAYS stay away from tanks engulfed in fire For massive fire,use unmanned hose holders
or monitor nozzles,if this is impossible withdraw from area and let fire bum.
6. ACCIDENTAL RELEASE MEASURES
e i Spas: Ensure adequate ventilation- Avoid contact with eyes or clothing E.acuate personnel to safe areas. Keep people away
from and upwind of spill/leak ELIMINATE all ignition sources(no smoking,flares,sparks.or flames in immediate area).
Do not touch damaged containers or spilled matenal unless weanng appropriate protective clothing. See section 8 for
more information. Stop leak if you can do it without risk. Use persona! protective equipment as required. Keep
combustibles(wood.paper,oil,etc)away from spilled material DO NOT GET WATER INSIDE CONTAINERS
Ventilate the area.Refer to protective measures listed in Sections 7 and 8.
Stop leak if you can do it without risk.Cover with DRY earth,DRY sand or other non-combustible material followed with
plastic sheet to minimize spreading or contact with rain
With dean shovel place matenal into clean.dry container and cover loosely:move containers from spill area.Flush area
with flooding quantities of water Prevent product from entering drains. Cover powder spill with plastic sheet or tarp to
minimize spreading and keep powder dry
7. HANDLING & STORAGE INFORMATION
-; Wait&t iyg rime 0r ;.-es Use personal protection equipment Avoid contact'with eyes, or clothing Keep away from heat hot surfaces, sparks,
open flames, and other ignition sources. No smoking Do not eat, dnnk, or smoke when using this product. Remove
contaminated clothing and shoes Use with local exhaust ventilation Handle in accordance with good industrial hygiene
and safety practice.Avoid formation of dust and aerosols.Avoid breathing dusts from this product.Avoid excessive dust
generation Further, processing of solid materials may result in the formation of combustible dusts The potential for
combustible dust formation should be taken into consideration before additional processing occurs. Provide appropriate
exhaust ventilation at places where dust is formed Use in a well-ventilated location(e.g.local exhaust ventilation,fans).
After use,wash hands and exposed skin with soap and water Do not eat,drink,or smoke while handling product
Storage.g-13ndmir Keep container tightly closed in a dry and well-ventilated place Keep this material away from heat. sparks, and open
flame Open containers slowly on a stable surface. Store containers in a cool, dry location, away from direct sunlight,
other light sources,or sources of intense heat.Store away from incompatible matenals(See Section 10).
Srw a Frecautoos. Storage class(TRGS 510):5.1 B:Oxidizing Materials

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SAFETY DATA SHEET BGT-005
Prepared to OSHA.ACC.ANSI,WHSR.WHMIS.2020/878/EU Standards I SDS Revision 2 2 I SDS Revision Date: 3/12/2024
8. EXPOSURE CONTROLS & PERSONAL PROTECTION
8' E.pos,re Limas AC H NOH C 1 OS OTHER
oc^' rym' ES- ES- ES-
CHEMICAL NAMOSI TLV STEL TWA STEL PEAK PEL STEL IOLH
SODIUM PERCARBONATE Nil NA NF NF NF NA NA NA
a 2 verrilatwn s Enpeeenry Use local or general exhaust ventilation to effectively remove and prevent buildup of dust or vapors generated from the
Cor""6 handling of this product.Ensure appropriate decontamination equipment is available(e.g.,sink,safety shower,eye-wash
station) Periodic medical exams of exposed workers may be required.
Pesp:ratay Protecaoe No protective equipment is needed under normal use conditions. If exposure limits are exceeded or
irritation is experienced,ventilation and evacuation may be required Keep the exposure within legal limits.
In the worker's breathing zone and the general area, dusts must be kept below the TLVs and the
equivalent exposure must compute to less than one. Keep exposure as low as possible. When dusts are
generated, and respiratory protection is needed, use only protection authorized by 29 CFR §1910.134.
applicable U.S. State regulations, or the Canadian CAS Standard Z94.4-93 and applicable standards of
Canadian Provinces,EC member States,or Australia
=4 Eye P-tre•a^-, Always use protective eyewear (e.g., tightly sealing chemical safety goggles) when handling, or when
cleaning spills or leaks. Contact lenses pose a special hazard: soft lenses may absorb and concentrate V
irritants. If necessary. refer to U.S. OSHA 29 CFR §1910.133. Canadian standards, or the European
Standard EN 166
Wear suitable, impervious gloves. Gloves must be inspected prior to use. Use proper glove removal
technique (without touching glove's outer surface) to avoid skin contact with this product Dispose of ID
contaminated gloves after use in accordance with applicable laws and good laboratory practices Wash and
dry hands.
5 So y Proter.thOn Wear suitable protective clothing Long sleeved clothing. Chemical resistant apron Wear fire/flame
resistant/retardant clothing. If necessary, refer to appropriate standards of Canada, the E C member
_states,or U.S OSHA
9. PHYSICAL & CHEMICAL PROPERTIES
9 1 Aottea'ance Solid.white powder
• 9-2 Odor Odorless
g.3 OdorTnreshc.ld NA
94 pH 10.5
9 s Meit ng Po-n<=r.e g Fort Decomposition at T'140°C
9 6 etat Bating Pantaaing NA
Range
g% Fnasnpant >230°C
9 e Jppertower Fiarranabiitr NA
Lrnurs
g g Vapor Pressure. NA
g.1C vapor Density. (25°C)negligible
9 1+ ?eLative Density 1.93 gfcm3
a 1: sauoirty Soluble in water.solubility 140 Q/L
9 13 Farman Coencrent isag P,,;' NA
9 14 Autogrvnon Temperature: NA
9 18 Demrrpastim Temperature. NA
91e visoosi,y. NA
g 17 Ot11er!^iamation. Bulk density.700-1200 kgtm3
10. STABILITY & REACTIVITY
'c 1 srabety: May cause fire or explosion,strong oxidizer
c` Hsrardais:Decar"p°s4w Oxides of carbon,oxygen
Product::
10.3 Harydoue Pdymerwaoon Spontaneous polymerization can occur.
104 Conditions to Avoao Exposure or contact to extreme temperatures.heat.sparks,flames,incompatible chemicals,moisture
-c a i"onmpatae Substances. Organic material.Combustible matenal Hydrocarbons Strong acids. Strong bases. Strong oxidizing agents.

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Page 4of6
SAFETY DATA SHEET BGT•005
Prepared to OSHA,ACC.ANSI,WHSR.WHMIS.2020/878/EU Standards I SDS Revision 22 I SDS Revision Date:31122024
11. TOXICOLOGICAL INFORMATION
1,I Rote co'Entry Inhaaucn I YES I A;7sapnon I YES 1 ingestion I YES
t1.2 ToxiciryData LDso (oral, mice). > 5009 1 ng/kg, ATE LD5. (oral. northern bobwhite) = 2472 mg/kg, Causes serious eye irntatot
(OECD Test Guideline 405).
11.3 Acute To'ccy: Senous rmtation to eyes.
114 Chronic Toxicity: This matenal may aggravate any pre-existing skin condition(e.g..demnatitis).
11.5 Suspected Carcinogen: No
11.6 Reprod4ctveToincity This product is not reported to produce reproductive toxicity in humans.
stutagenrny. This product is not reported to produce mutagenic effects in humans.
Errnryotoxicty This product is not reported to_produce embryotoxic effects in humans.
Terxogen.oty. This product is not reported to cause teratogenic effects in humans
117 ;rrnancyo'Product General Nuisance Dusts. Nuisance dusts,which are essentially nontoxic and chemically non-irritating. Skin contact has
shown no problems other than possible drying and mechanical irritation. Eye contact can produce particulate irritation
Excessive inhalation can produce mild pulmonary irritation and possible non-disabling slight fibrosis of the lungs. The
product can cause allergic skin reactions(e.g,rashes,welts,dermatitis)upon prolonged or repeated exposure
., 5iO4o124ai Exposure indices NE
'1-rsician RecommendaWns Treat symptomatically.
12. ECOLOGICAL INFORMATION
,z 1 Enorvrxneatai Stability There is no specific data available for this product.
121 Eifece an iota tis&Anrtnais: Honey Bees (Apis mellifera)' At 48 hours, mortality in the control and test groups was 0% (Table 1). Because the
mortality of the test group was the same as the control group mortality(0%),there is no difference and the LD50 of Lake
Guard Oxy is considered greater than 100 lig a.i./bee(OECD Guideline 213)
Effete on Aquatic Life ECsc.(48h,static acute,Cenodaphnia dubia.water flea):>37.5 mg/L. Test EPA-821-R-02-012(2002)Method 2002.0
LCff,(Zebra fish),96h): > 100.0 mg/L,WGK i (slightly hazardous to water)
13. DISPOSAL CONSIDERATIONS
13.1 waste D,sposa/ Disposal should be in accordance with local,state,and federal regulations Dispose of in accordance with federal,state,
and local regulations.
Special Considerations. ' Offer surplus and non-recyclable solutions to a licensed disposal company Contact a licensed professional waste
disposal service to dispose of this material_
14. TRANSPORTATION INFORMATION
The basic description(ID Number,proper shipping name hazard class 8 division,packing group)is shown for each mode of transportation Additional descriptive information
may be required by 49 CFR,IATAiiCAO.IMDG and the CTDGR.
4 1 4P CFR(GNM: UN3378,SODIUM CARBONATE PEROXYHYDRATE,5.1,Ill,LTD QTY(IP VOL s 5.0 kg)
14: IATA(AWN: UN3378,SODIUM CARBONATE PEROXYHYDRATE,5.1,III,LTD QTY(IP VOL s 1.0 kg)
14.3 WOG(OCN). UN3378,SODIUM CARBONATE PEROXYHYDRATE,5.1,III,LTD QTY(IP VOL s 5.0 kg)
144 TDGR(Canadian GND) UN3378,SODIUM CARBONATE PEROXYHYDRATE,5 1,III,LTD QTY(IP VOL s 5.0 kg)
14.5 ADRtRiD 1Eu1: UN3378,SODIUM CARBONATE PEROXYHYDRATE,5.1,III,LTD QTY(IP VOL s 5.0 kg) 040Je1
14 a 3CT(MEr1Cor UN3378,CARBONATO DE SODIO PEROXIHIDRATADO,5 1,III,CANT.LTDA (IP VOL s 5.0 kg) S 1
14- ADGR tAusi UN3378,SODIUM CARBONATE PEROXYHYDRATE„5 I,III,LTD QTY(IP VOL s 5.0 kg)
14 a u.S CENS4-6:FORE't3N SCHEDULE B.2836.99.0000 or 2836 99.0007
TRADE
15. REGULATORY INFORMATION
1 SARA Repotting This product does not contain any substances subject to SARA Title III.Section 313 reporting requirements.
Ragureenerrs.
15: SARA TP- There are no specific Threshold Planning Quantities for the components of this product
15 3 TSCA inventory Status The components of this product are listed on the TSCA Inventory
154 CERCLA Reportable NA
Quantity
-
15 otner Feoerai Regderemens: NA
15 a Omer Canadian Regdations This product has been classified according to the hazard criteria of the Controlled Products Regulations(CPR)and the
SDS contains all the information required by the CPR. The components of this product are listed on the DSL/NDSL.
None of the components of this product are listed on the Pnonties Substances List
15 7 state RoguiatorY lnrormauoa No ingredients in this product, present in a concentration of 1.0% or greater, are listed on any of the foi)owmg state
criteria lists.California Proposition 65(CA65).Delaware Air Quality Management List(DE).Florida Toxic Substances List
(FL). Massachusetts Hazardous Substances List (MA), Michigan Critical Substances List (MI), Minnesota Hazardous
Substances List(MN), New Jersey Right-to-Know List(NJ), New York Hazardous Substances List(NY), Pennsylvania
Right-to-Know List(PA),Washington Permissible Exposures List(WA),Wisconsin Hazardous Substances List(WI).
_ Renu,remens NA

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Page 5 of 6
SAFETY DATA SHEET BGT-005
Prepared to OSHA,ACC,ANSI.WHSR,WHMIS.2020l878/EU Standards I SDS Revision 2.2 I SDS Revision Date:3/122024 _
16. OTHER INFORMATION
ie t der irlonn3ton DANGER! MAY INTENSIFY FIRE - OXIDIZER. MAY BE HARMFUL IF SWALLOWED. CAUSES SERIOUS EYE
DAMAGE.
Keep away from heat, hot surfaces, sparks, open flames, and other ignition sources. No smoking. Keep
away from clothing and other combustible materials.Wash hands and exposed skin areas thoroughly with
soap and water after handling. Do not touch eyes. Wear protective gloves/ protective clothing/ eye
protection/ face protection. IF SWALLOWED: Get emergency medical help. IF IN EYES: Immediately rinse
with water for several minutes. Remove contact lenses if present and easy to do - continue rinsing. Get
medical help. In case of fire:Use dry sand,dry chemical,or alcohol-resistant foam for extinction.
KEEP OUT OF REACH OF CHILDREN.
2 -gym&Gem. See last page of this Safety Data Sheet
.e 3 Dmciairner This Safety Data Sheet is offered pursuant to OSHA's Hazard Communication Standard, 29 CFR §1910.1200. Other
regulations must be reviewed for livability to thisproduct.To the best of Shi Mate's&BlueGreen Water
governmentpp p
Technologies Ltd.'s knowledge. the information contained herein is reliable and accurate as of this date; however,
accuracy.suitability or completeness is not guaranteed and no warranties of any type, either expressed or implied,are
provided. The information contained herein relates only to the specific product(s) If this product(s) is combined with
other materials, all component properties must be considered Data may be changed from time to time Be sure to
consult the latest edition.
' Pr'✓''e•, BlueGreen Water Technologies Ltd.
16 HaMtktsoot Blvd
Modfin-Maccabim-Re'ut,Israel 7178096
Tel:[phone]
Fax:[phone]
Email:info©bgtechs.com
https llbgtechs corn!
e: Pr�e-ea 5r ShipMate.Inc.
PO.Box 787
Sisters,Oregon 97759-0787 USA ShipMate
Tel:[phone]
{ )
Fax:[phone]
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SAFETY DATA SHEET BGT-005
Prepared to OSHA,ACC,ANSI,WHSR.WHMIS.2020/878/EU Standards I SOS Revision.2 2 I SDS Revision Date:311212024
DEFINITION OF TERMS
A.;arge number of abere,aoons to acronyms aoper on a SOS.Some of these:hat are commonly;used Include one beoamg:
GENERAL INFORMATION: NATIONAL FIRE PROTECTION ASSOCIATION:NFPA
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pa Pemrissbe Exposure Land 0 Min,mat-1,azagd F[aWAYm
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iLV 'hreshold LirM L'aL.e 2 Moderate Hazard
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Study Title
Acute Toxicity of Lake Guard Oxy from
BlueGreen Water Technologies
To the Water Flea(Ceriodaphnia dubia)
Under Static Test Conditions
Performed For
BlueGreen Water Technologies
301 South Hills Village Ste LL200#452
Pittsburg,PA 15241
Project Officer
Jessica Frost
Author
Janelle Mikulas,M.S.
Study Period
03 February 2023 to 05 February 2023
Performing Laboratory
STILLMEADOW
INCORPORATED
Environmental Toxicology Laboratory
12852 Park One Drive
Sugar Land,Texas 77478
Telephone: [phone]
Fax: [phone]
fT
Certificate Number T1 047043 52-22-1 5
Project Numbers
23-613-002

•
STATEMENT OF PROCEDURAL COMPLIANCE
I certify that this document and all attachments were prepared under my direction or supervision in
accordance with a system designed to assure that qualified personnel properly gather and evaluate the
information submitted: information contained herein is accurate and complete.
rJ
J�
Ja ikulS. Date
STATEMENT OF QUALITY ASSURANCE
The report and study data were audited to assure that the study was performed in accordance with
STILLMEADOW, Inc. Standard Operating Procedures and regulatory guidelines. This report is an
accurate reflection of the raw data.
Quality Assurance Auditor Date
23-613-002 BlueGreen Water Technologies
Page 2

EXECUTIVE SUMMARY
The objective of this study was to determine the acute toxicity of Lake Guard
Objective Oxy samples submitted by the BlueGreen Water Technologies to
Ceriodaphnia dubia.
Study Director Janelle Mikulas,M.S.
Test Type 48-Hour Static Acute
United States Environmental Protection Agency(EPA-821-R-02-012)(2002)
Test Method Method 2002.0 (Ceriodaphnia dubia).
Test
Dates (Times) 03 February 2023 (1220) to 05 February 2023 (1120)
Test Substance Lake Guard Oxy
Control/Dilution Receiving Water
Water
Test 15mg/L,30mg/L,45mg/L,C2
Concentrations
Source of Organisms STILLMEADOW,Inc. Culture Laboratory
Age of Organisms <24 hours
Test Test Organism Test Data EPA Criterion
Acceptability C. dubia 100% >_90%
(Control Survival)
Test Organism NOEC' LOEC2 IC50
Test Results
C. dubia 30 mg/L 45 mg/L 37.5 mg/L
'NOEC=No Observed Effect Concentration
2LOEC=Lowest Observed Effect Concentration
23-613-002 BlueGreen Water Technologies
Page 3

INTRODUCTION
The objective of this study was to determine acute toxicity to Ceriodaphnia dubia of Lake Guard Oxy
sample submitted by BlueGreen Water Technologies. This study is conducted in compliance with Texas
Pollution Discharge Elimination System (TPDES) permit requirements; and in accordance with Texas
Water Code Chapter 5, Subchapter R, Title 30 Texas Administrative Code Chapter 25 and the National
Environmental Laboratory Accreditation Program (NELAP), Certificate Number T104704352-22-15.
All original data, laboratory notebooks, and associated documentation are archived by the
STILLMEADOW, Inc. Environmental Toxicology Laboratory.
METHODS AND MATERIALS
Test Substance
One product sample provided by the sponsor as Lake Guard Oxy was used for testing.
Dilution Water
Dilution water was receiving water collected and prepared according to USEPA (2002) guidelines. An
additional control group was conducted concurrently consisting of 100% moderately hard synthetic
freshwater, designated C2. Initial characterization of the dilution and control water is given in Table 2.
Table 2. Initial water quality parameters of the control and dilution water tested.
Alkalinity Hardness Ammonia Total
Residual
P Sample
Batch/Sample# pH Conductivity
Receiving Water Collection (SU) CaCO) Ca O) (µmhos/cm) (3 Chlorine
(ma/L)
Q230021.01 (�6 Jan 23
145 EST) 7.3 30 165 620 1.07 0.621
Batch# Total
Alkalinity Hardness Ammonia
Date pH Conductivity Residual
Moderately Prepared (SU) (µmhos/cm)(mg/L OWLg/L m L
Hard SyntheticCaCO3) CaCO3) ( Chlorine
Freshwater N1 ) (mg/L)
QA23013 27 Jan 23 7.8 58 94 321 0.01 0.01
'Out of range
23-613-002 BlueGreen Water Technologies
Page 4

Procedures
Testing was carried out according to procedures specified in Methods for Measuring the Acute Toxicity of
Effluents and Receiving Water to Freshwater and Marine Organisms (EPA, 2002). The test organisms
were less than 24-hour-old C. dubia cultured at STILLMEADOW, Inc. In the test, 40 organisms (5
replicates of 8 organisms each) were exposed to each test concentration. Organisms were not fed during
the test.
The test was performed using moderately hard synthetic freshwater as an additional control (C2) and
receiving water as a control and diluent. The effluent concentrations tested were 15, 30 and 45 mg/L.
Temperature, pH,conductivity,and dissolved oxygen were measured before the addition of the organisms.
Aeration was not employed.
The test was terminated after 48±1 hours and a count of the surviving organisms was recorded. Dissolved
oxygen,conductivity,pH,and temperature in the old test solutions were also recorded.
DATA ANALYSIS
All data were analyzed according to the statistical flow charts outlined in the EPA acute testing manual
(USEPA 2002). Table 3 lists the methods that were used in the survival data analyses. ToxCalc® was
used for all hypothesis-testing evaluations. If a dose response occurred, ToxCaic® was used for point-
estimation techniques. All printouts of statistical results are included in Appendix B.
Table 3. Statistical methods used to analyze data for the toxicity test.
Endpoint Comparison Procedure
Transformation Arc Sine(y)'2
Normality Shapiro-Wilk's Test(aS0.01)
48-Hour NOEC Survival Homogeneity of Variances Cannot Be Confirmed
Reduction Relative to Control Steel's Many-One Rank Test
(a=0.05)
48-Hour LC50 Survival Not Applicable Linear Interpolation
23-613-002 BlueGreen Water Technologies
Page 5

RESULTS
Ceriodaphnia dubia
Survival in the receiving water control met EPA test acceptance criterion for acute toxicity test (z90%).
(Table 5).
Table 5. Percent survival of Ceriodaphnia dubia in the 48-hour toxicity test
Percent Survival
Time Replicate Omg/L 15mg/L 30mg/L 45mg/L C2
A 100 100 100 0 100
B 100 100 100 0 100
48 Hours C 100 100 100 0 100
D 100 100 100 0 100
E 100 100 100 0 100
48-Hour Mean _ 100 100 100 0 100
Standard Deviation 0.0 0.0 0.0 0.0 0.0
Coefficient of Variation(CVmg/L) 0.00 0.00 0.00 --- 0.00
STANDARD REFERENCE TOXICANT TESTS
STILLMEADOW, Inc. conducts routine standard reference toxicant testing using Ceriodaphnia dubia
obtained from STILLMEADOW, Inc. cultures. Sodium Chloride is used as the reference toxicant with
moderately hard synthetic freshwater as the dilution water; the test method followed is USEPA (2002).
A copy of STILLMEADOW, Inc.'s most recent standard reference toxicant control chart for this species
is presented in Appendix C.
STUDY DEVIATIONS
Total residual chlorine was out of range in sample Q230021.01. No other deviations from the prescribed
guidelines or standard operating procedures were identified during the study.
REFERENCES
EPA, 2002. Methods.for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater
and Marine Organisms, Fifth Edition, October 2002. EPA-823-R-02-012. Environmental
Monitoring and Support Lab. Cincinnati,Ohio. Fourth Edition. 293 pp.
Ives, Michael A. TOXCALCTM Version 5.0. 1994. TidePool Scientific Software. McKinleyville,California.
23-613-002 BlueGreen Water Technologies
Page 6

APPENDIX A
Chain of Custody,Check-In,and Chemistry Forms

CHAIN OF CUSTODY RECORD
ClientlPmject Name y Project Location
ee r"G to (� /"t mA tf is 1/cr-- ANALYSES
Project No. Field Logbook No.
�-3- 613- oar Sampler(Signature)(Signature) Chain of Custody Tape No.
Sample No./ Start Start Finish Finish Lab Sample Type of
Identification Date Time Date Time Number Sample REMARKS
- latiy I1.-i C.Sf Qa30o2l.0j
Relinquished by:(Signature) Date Tune Received by:(Signature) Date Time
Relinquished by:(Signature) Date lime Received by:(Signature) Date Time
Relinquished by:(Signature) Date Time Received by:(Signature) Date lime
Sample Disposal Method: Disposed of by:(Signature) Date Tune
ANALYTICAL LABORATORY
SnIIIiIjI111\4I .L\J)OW Environmental En�2��O �ebogg8
yy Laboratory
INCORPORATED 281-240-8828

AQR&I0999.02 page 1 of 1
STILLMEADOW, Inc. ENVIRONMENTAL TOXICOLOGY LABORATORY
Test Reference & Control Substance Receipt& Inventory Log
Project Number: L�9 3- u 15 - (7J 1 ! vt>2. Sample No.: a a3 DO•!. a
Substance Name: I U/ GvtKc J IA .yr+k.rker+Sponsor: 8/1, e 6"eeN
Description: L! Lr d No./Size Containers: O,✓e- V Z.Amount Received: / /i. Lot/Batch/Set No. '
Received By: rfr 7 Date/Time Received: / 3 7 D .27 (779r 43
Received From: Gri e.,--/• Condition: 6 so d
Shipper: STILLMEADOW,Inc.
PP Odisii) ci!�i (circle) Bill of Lading No.:
---
Storage Requirements: L
g �I 0�G Storage Location: 4--/
Comments: .----
Parameters Upon Arrival: / $-'fir' Init./Date: ,4' .27 T',- 2-3
Salinity(ppt): D.3 0 Meter#: 70 5-
PH(SU): 7 3 Meter#: 1/2
Temp(°C): 6 Meter#: rZo2
Refrigerant: Fro a Thawed None (circle) Ice Ca or Igloo Ice (circle)
DO(mg/L): 6 7 Y Meter#: S-6 Y
Cond.(µmhos/cm): 6 a 0 Meter#: 7 a f
Initial TRC: .—
Meter#: 7D 6 Reagent Lot#: Exp.
Amount Na2SO3 Added: -___"`�
Post Dechlorination TRLV0 Meter#:
Chemistry Sample List: Y oteRcircle)
Black Spots: Y orr(circle)
Foul Smell: Y or (circle)
OSG&Block: ....---
Preserved with Nitrogen: Y orrcircle)
Notes:
Si ature: � � s�
(A) IV)(In
' ' "" °-I cc,b0,1>Ait_ �n�crS 4 6Ui ivy , ,���
��'\ S nip 6 L ,M (, od lry/
STILLMEADOW,Inc.
Revision Date: August 2011

I
Sample Wet Chemistry Page i
Study: )7) - L I ) `3 0}1/3(;)\ Results To: A
Alkalinity `� Hardness .) Ammonia Nitrites Nitrates
Alkalinity(mg/L CaCO3)-pH meter#: ' 0,
Aliquot Titrant(mL) Dilution
Sample (mL) [0.02 N H2SO4] Factor Alkalinity Initials Time Date
Man./Lot#:1:,A,a./Milli"'
ga304,),I.1 :-)-‘(3 I ; :'h J 3 flf-) f',o, sgi7-c6Z
Hardness(mg/L CaCO3)—Water Hardness Buffer Man./Loth:1'„Q M•6‘"1-1-11/lndicator Man./Lot#: ',(,S/No y),o,,i),
Aliquot Titrant(mL) Dilution
Sample (mL) [0.01 M EDTA) Factor Hardness Initials Time Date
Man./Lot#:1•.1)a/1",0501
Ammonia (mg/L NH3N)—Meter#: 1 ,n I/
Salicylate Reagent Man./Lot#: IA hM N/A .";.A Cyanurate Reagent Man./Lot#: 1-4 acKd4i�c i
Sample Aliquot Initial Dilution Ammonia Initials Time Date
(mL) Concentration Factor
Nitrites(mg/L)—NitriVer 3 Nitrite Reagent AccuVac Ampuls Lot#:
Aliquot Conversion
Sample (mL) NO2-N Factor NO2- Initials Time Date
Nitrates(mg/L)—NitraVer 5 Nitrate Reagent AccuVac Ampuls Lot#:
Aliquot Conversion NO Initials Time Date
Sample (mL) NO3 N Factor s . .
Calculations Check: 4k a L 7
AQN/N0612.03 STILLMEADOW, Inc.
Revision Date: Sept 2012

APPENDIX B
Statistical Analyses

Ceriodaphnla Survival and Reproduction Test-48 Hr Survival
Start Date: 2/3/2023 Test ID: 23-613-002 Sample ID: Lake Guard Oxy
End Date: 2/5/2023 Lab ID' Q2300224.01,> Sample Type: product
Sample Date: Protocol: EPA-821-R-02-012 Test Species: CD-Ceriodaphnia dubia
Comments:
Conc-mg/L 1 2 3 4 5
control 1.0000 1.0000 1.0000 1.0000 1.0000
C2 1.0000 1.0000 1.0000 1.0000 1.0000
15 1.0000 1.0000 1.0000 1.0000 1.0000
30 1.0000 1.0000 1.0000 1.0000 1.0000
45 0.0000 0.0000 0.0000 0.0000 0.0000
Transform:Arcsin Square Root Rank 1-Tailed Isotonic
Conc-mglL Mean N-Mean Mean Min Max CV% N Sum Critical Mean N-Mean
control 1.0000 1.0000 1.3931 1.3931 1.3931 0.000 5 1.0000 1.0000
C2 1.0000 1.0000 1.3931 1.3931 1.3931 0.000 5
15 1.0000 1.0000 1.3931 1.3931 1,3931 0.000 5 27.50 18.00 1.0000 1.0000
30 1.0000 1.0000 1,3931 1.3931 1.3931 0.000 5 27.50 18.00 1.0000 1.0000
45 0.0000 0.0000 0.1777 0,1777 0.1777 0.000 5 0.0000 0,0000
Auxiliary Tests Statistic Critical Skew Kurt
Shapiro-Wilk's Test indicates normal distribution(p>0,01) 1 0.835
Equality of variance cannot be confirmed
The control means are not significantly different(p = 1.00) 0 2.306
Hypothesis Test(1-tail,0.05) NOEC LOEC ChV TU
Steel's Many-One Rank Test 30 45 36.7423
Linear Interpolation (200 Resamples)
Point mg!L SD 95%CL(Exp) Skew
IC05 30.750 0.000 30.750 30.750 #DIV/0!
IC10 31.500 0.000 31.500 31.500 #DIV/0!
IC15 32.250 0.000 32.250 32.250 #DIV/0! 1.0
IC20 33.000 0,000 33.000 33.000 #01V/01
IC25 33.750 0.000 33.750 33.750 #DIV/0! 0.9 .
IC40 36.000 0.000 36,000 36.000 #DIV/0! 0.8 -
1050 37.500 0.000 37.500 37.500 #DIV/0I 0 7 -
a 0.6 -
c
a 0.5-
0.4 -
re
0.3 -
0.2 -
0.1 -
0.0♦ 4
0 10 20 30 40 50
Dose mg/L
Page 1 ToxCalc v5.0.23 Reviewed by' J �
--104A/9221
(rYu0- --L.x tI.I. ^N et9.47,

Ceriodaphnia Survival and Reproduction Test-24 Hr Survival
Start Date: 2/3/2023 Test ID: 23-613-002 Sample ID: Lake Guard Oxy
End Date: 2/5/2023 Lab ID: Q23002P.01 Sample Type: product
Sample Date: Protocol: EPA-82'I-R-02-012 Test Species: CD-Ceriodaphnia dubia
Comments:
Conc-mg/L 1 2 3 4 5
control 1.0000 1.0000 1.0000 1.0000 1.0000
C2 1.0000 1.0000 1.0000 1.0000 1.0000
15 1.0000 1.0000 1.0000 1.0000 1.0000
30 1.0000 1.0000 1.0000 1.0000 1.0000
45 0.0000 0.0000 0.0000 0.0000 0.1250
Transform: Arcsin Square Root Rank 1-Tailed Isotonic
Conc-mglL Mean N-Moan Mean Min Max CV% N Sum Critical Mean N-Mean
control 1.0000 1.0000 1.3931 1.3931 1.3931 0.000 5 1.0000 1.0000
C2 1.0000 1.0000 1.3931 1.3931 1.3931 0,000 5
15 1.0000 1.0000 1.3931 1.3931 1.3931 0.000 5 27.50 17.00 1.0000 1.0000
30 1.0000 1.0000 1.3931 1.3931 1.3931 0,000 5 27.50 17.00 1.0000 1.0000
*45 0.0250 0.0250 0.2144 0.1777 0.3614 38.301 5 15.00 17.00 0.0250 0.0250
Auxiliary Tests Statistic Critical Skew Kurt
Shapiro-Wilk's Test indicates non-normal distribution(p<=0.01) 0.4971 0.868 3.24893 13.4118
Equality of variance cannot be confirmed
The control means are not significantly different(p= 1.00) 0 2.306
Hypothesis Test(1-tail, 0.05) NOEC LOEC ChV TU
Steel's Many-One Rank Test 30 45 36.7423
Linear Interpolation(200 Resamples)
Point mg/L SD 95%CL(Exp) Skew
IC05 30.769 0.020 30.744 30.823 0.9834
IC10 31.538 0.040 31.488 31.647 0.9834
IC 15 32.308 0.060 32.233 32.470 0.9834 1.0
IC20 33.077 0.080 32.977 33.293 0.9834 -
1C25 33.846 0.099 33.721 34.116 0.9834 0.9 ,
1C40 36.154 0.159 35.954 36.586 0.9834 0.8-
IC50 37.692 0.199 37.442 38.233 0.9834 0.7
0.6-
c
00,5-
N
a) 0.4-
0.3-
0.2 -
0.1
0.0♦
0 10 20 30 40 50
Dose mg/L
Page 1 ToxCalc v5.0.23 Reviewed bye i(�
'3,v+-. ^ ►..��

APPENDIX C
Standard Reference Toxicant Control Charts

Ceriodaphnia dubia Short-Term Chronic Standard Reference Toxicant Control Charts
LC25 (mg/L NaCI) Survival
3500.00 1
3000.00 - •
- 2500.00 — •
Z 2000.00 --— -
J )(
1500.00 — ■ LC25 Survival
Upper Limit
ID 1000.00
N �---Lower Limit
U 500.00 x Mean LC25
J
0.00
-500.00
�-- NNNNNNNNNN N N
NNNNNNNNNNN N N
0 Co 0 0 0 0 0 0 0 O 0 0 0
N NNNNN N NNNN N N
O O CO CO co O co 'et co co O 'a'
N N a- N r N r- N N
N - N C7 et 11) LC/ ti Oo Cr) O a-
IC25 (mg/L NaCI) Reproduction
2500
■
2000
cvv▪ 1500
Z
—J • ■ IC25 Growth
c) 1000 N — x iG •
-_-X—lt x
i
-Upper Limit
X
Lp •
Lower Limit
N 500 — — — —
. x Mean IC25
0
-500 • NNNNNNNN N NNNN
NNNNNNNNNNNN N
0 0 0 0 O 0 0 0 0 0 0 0 0
N N N N N N N N N N N N N
O 0 Co co Co co co d' co co O
• N N s- N r- N r- a- N N
N N c") 'et to tD t- CO C) 0 a-' e-
1 J
STILLMEADOW
INCORPORATED 2/20/2023

STILLMEADOW
INCORPORATED
TITLE
Lake Guard Oxy
Honey Bee,Apis mellifera, Acute Oral Toxicity Limit Test
TEST GUIDELINE
OECD 213
•
AUTHOR
Cole Younger,PhD
STUDY COMPLETION DATE
11 August 2023
PERFORMING LABORATORY
STILLMEADOW,Inc.
12852 Park One Drive
Sugar Land,TX 77478
LABORATORY STUDY ID
26312-23
PAGE 1 of 29
12852 Park One Drive ■ Sugar Land, Texas 77478 IN 281 240-8828 ■ Fax [phone]
www.stillmeadow.com

26312-23
Page 2 of 29
GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT
The following is a detailed description of all differences between the practices used in the study and
those required by United States Environmental Protection Agency FIFRA 40 CFR Part 160:
Section 160.31 (d)and 160.105(a)(b)(e)Characterization and stability information was not provided
to the testing facility.
Section 160.113(a)Mixture analysis was not performed.
The following is a detailed description of all differences between the practices used in the study and
those required by Organization for Economic Cooperation & Development Principles of GLP,
ENV/MC/CHEM(98) 17:
Section II,1.1(2)(p),6.1(1)and 6.2(2)(4)Characterization and stability information was not provided
to the testing facility.
Section II,6.2(5)Mixture analysis was not performed.
Study ///2/6 2-3
Director: /,� Date:
�`c/
Cole Younger,PhD
STILLMEADOW,Inc.
Sponsor: ` Date: " 'Z3
Name of Sig r e.Ssi cc. FlidDlr
Sponsor: BlueGreen Water Technologies
Submitter: '—y e� `� Date: i u� L141202-3
Name of Signer:(,7/J ASS i ca. 1 v i'
Submitter: BlueGreen Water Technologies
STILLMEADOW,Inc.

26312-23
Page 3 of 29
QUALITY ASSURANCE STATEMENT
Study Title: Lake Guard Oxy
Honey Bee,Apis mellifera,Acute Oral Toxicity Limit Test
The study report and data have been audited in accordance with Good Laboratory Practice Standards
and STILLMEADOW, Inc., Standard Operating Procedures (SOPs). The final report accurately
reflects the study data. The Quality Assurance Unit has not been involved in the actual conduct of this
study.
The Quality Assurance Unit performed a facility inspection on 25 Apr 23. All findings were reported
to Management,and the report and responses are kept in the Quality Assurance files.
The findings from any study inspections and audits were reported to the Study Director and
Management as follows:
Critical Phase Inspected Date Inspected Reported to Reported to
Study Director Management
Protocol Review 08 May 23 08 May 23 08 May 23
Observations 07 Jun 23 07 Jun 23 07 Jun 23
Report!Data Audit 22 Jun 23 22 Jun 23 22 Jun 23
44
Kristina Rodrigue,RQ LP Date
Director,Quality Assurance Unit
STILLMEADOW,Inc.
STILLMEADOW,Inc.

i
I

26312-23
Page 4 of 29
TABLE OF CONTENTS
Page
GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT 2
QUALITY ASSURANCE STATEMENT 3
SUMMARY 5
INTRODUCTION 5
SPONSOR INFORMATION 6
TEST ITEM 6
CONTROL SUBSTANCE 6
TEST SYSTEM 7
Insect Species 7
Insect Husbandry 7
PROCEDURES 8
Preparation of Test Bees 8
Dose Calculation and Preparation of Test Item 8
Preparation of Sucrose Solution 8
Preparation of Positive Control 8
Test and Control Substance Administration 8
Observations 9
Test Validity 9
LDso for Positive Control 9
Statistical Analysis 9
Evaluation of Results 9
RESULTS AND DISCUSSION 10
Data Not Used 10
Protocol Deviation 10
Test Validity 10
Control. Test item and Toxic Standard Consumption 10
LD50 for Positive Control 10
Clinical Signs of Toxicity, Mortality Observations and Evaluation 11
CONCLUSION 12
SIGNATURE 12
STUDY PERSONNEL 12
TABLES AND APPENDICES
Table I -Mean Number of Dead Honey Bees and Percent Mortality Summary 11
Table 2 - Food Consumption 13
Table 3 -Mortality, Observations and Percent Mortality 14
Appendix A -Signed Protocol 15
Appendix B-Sponsor Provided Product Label 21
Appendix C- Positive Control Certificate of Analysis 28
Appendix D- LD50 Analysis for Positive Control 29
STILLMEADOW.Inc.

26312-23
Page 5 of 29
SUMMARY
This study was designed to assess the acute oral toxicity potential of the test item, Lake Guard Oxy,
when administered in the diet of the adult worker honey bee, Apis mellifera, at 100 jig active
ingredient(a.i.)/bee. The test item and toxic standard were administered orally in a 50%sucrose
solution to honey bees for four hours,with a control group fed only sucrose solution. Observations for
mortality and clinical signs of toxicity were made at approximately 4, 24, and 48 hours after dosing in
the control group,test group,and positive control groups(0.025,0.25 and 2.5 jig/bee dimethoate).
Bees were offered 100 µL of the test substance solution with a nominal concentration of 20 µg
For a total concentration of 2000 jig a.i./l00 µL,giving a nominal concentration of 100 µg a.i./bee in a
cup of 20 bees.
The estimated Median Lethal Dose(LD50) for dimethoate at 24 hours was 0.307 jig/bee, which falls
within the validity requirement of 0.10-0.35 jig/bee.
At 48 hours, mortality for the control group, test group, and positive control groups (0.025, 0.25 and
2.5 jig/bee dimethoate), was 0%, 0%, 2%, 41%and 100%, respectively. The control and test groups
consumed an average of 0.119 g and 0.117 g per replicate,respectively,from the 100 jiL offered of the
appropriate substance during the study. The validity requirement of no more than 10%control mortality
was met with a control mortality of 0%. The test group mortality at 0%did not exceed control mortality,
therefore,the difference was not significant and the LD5o of Lake Guard Oxy is considered greater than
the nominal concentration of 100 jig a.i./bee.
INTRODUCTION
The objective of this study was to assess the acute oral toxicity potential of the test item, Lake Guard
Oxy, when administered orally to honey bees as part of their diet in accordance with Organization for
Economic Cooperation & Development Principles (OECD) Guideline 213. The study was also
conducted according to the approved protocol (included as Appendix A)and STILLMEADOW, Inc.,
SOPs. There were no deviations from the protocol that affected the quality or outcome of the study.
The study was initiated on 11 May 23, and the laboratory portion of the study was conducted from
06-08 Jun 23. All raw data, original protocol,original final report,any amendment(s),and a retained
test item sample are archived at STILLMEADOW,Inc. for a period of 15 years.
STILLMEADOW.Inc.

26312-23
Page 6 of 29
SPONSOR INFORMATION
Company Name: BlueGreen Water Technologies
Address: 3101 South Hills Village Ste LL200#452
Pittsburgh, PA 15241
TEST ITEM
Reference Identification: Lake Guard Oxy
Label Identification: BlueGreen WATER TECHNOLOGIES Lake Guard Oxy EPA Reg No.
93647-2
At Sponsor request,a copy of the Sponsor provided product label is included as Appendix B.
Date and Quantity Received: 11 May 23;332.8 g(GW)
Physical Description: White pellets
Storage: Room temperature
Purity&Composition: Not provided to testing facility
P
Stability: Not provided to testing facility
for characterization and stability,and the level of GLP compliance for that data,are the
Datagenerated ,
h' P
responsibility of the Sponsor. Records pertaining to identity, synthesis methods and location of
documentation are the responsibility of th a Sponsor.
CONTROL SUBSTANCE
Positive control
(toxic standard): Dimethoate(0.025,0.25 and 2.5 pg/bee)
CAS#60-51-5
(Mfr:Chem Service.Lot: 13914600,Exp: 31 Mar 2025)
The manufacturer's Certificate of Analysis for the positive control is included as report Appendix C.
STILLMEADOW,Inc.

26312-23
Page 7 of 29
TEST SYSTEM
Insect Species
Species/Strain/Source: Apis mellifera/Italian honey bee/STILLMEADOW, Inc.bee colony,
disease and pest-free with no previous pesticide exposure.
Justification of Species: The honey bee is the species required in the regulatory guidelines for
this study.
Quantity and Age on Day 0: 500 bees; 100 bees per group in five replicates of 20 bees each
Identification: Numbered cups with treatment identification
Acclimation: Acclimation is not applicable for honey bees.
Insect Husbandry
Housing: Disposable 16 ounce paper cup with a screen lid with 20 bees per cup.
Environmental Controls
Set to Maintain: Temperature 33 ±5°C Relative Humidity 50-70%
• Dark except when dosed or observations were made
Actual Temp/Rel. Humidity: 32°C/53 -55%
Handling: Only as much handling as necessary to conform to the test procedures
was allowed. The bees were shielded from excessive activity or other
disturbance during holding and testing.
Food/Source: 50:50 w/v: sucrose: dechlorinated (DC) water solution fed ad lihiium
after dosing period (sucrose: Mfr: Sigma Aldrich, Lot: SLC2154,
Exp: Apr 2027)
No contaminants were expected to have been present that would have interfered with or affected the
results of the study.
STILLMEADOW.Inc.

26312-23
Page 8 of 29
PROCEDURES
Preparation of Test Bees
Bees were fasted for approximately two hours before dosing.
Dose Calculation and Preparation of Test Item
To prepare a 20 µg a.i./µL solution, 0.20 g of test substance was dissolved into 10 mL of 50%sucrose
solution. The dosing solution was prepared according to the following calculation and was based on
100%test item concentration:
2000 pg 100,uL * 10(convert to mL) 20000 jug mL
20000 pg niL 1, 000, 000(convert to g) — 0.02 g'mL
Solution will he prepared in 10 mL; therefore 0.020 g x 10 mL —
0.20 g(200 mg)for every 10 ml.of 50%sucrose solution prepared
The solution was allowed to sit for approximately 1 hour before dosing to allow the active substance to
be released from the coating.
Preparation of Sucrose Solution
The 50%sucrose solution was prepared by dissolving 500 g of sucrose in 500 mL of sterile DC water.
Solution was prepared in the same manner as needed to be provided ad libitum.
Preparation of Positive Control
On day 0, three dose levels (0.025, 0.25, and 2.5 µg/bee) of the toxic standard, dimethoate, were
prepared by bringing 0.005 g of dimethoate to 10 mL with the 50% sucrose solution to make a
0.5 mg/mL stock solution,which was serially diluted to make 0.05 mg/mL and 0.005 mg/mL solutions.
The positive control solutions were dosed at 100 µL per cup.
Test and Control Substance Administration
On day 0, the honey bees were immobilized using CO2, randomly taken from the test container and
divided into five groups with five cups of 20 bees in each group. Group I was provided only the 50%
sucrose solution and served as the untreated control group. Bees in Group II were given the test item
at 2000 µg a.i./100 µL to equal 100 µg a.i./bee. Bees in Groups III-V were dosed with the toxic
standard,dimethoate,at 0.025,0.25,and 2.5 µg/bee,respectively,and served as the positive control.
For dosing, a micropipette tip containing 100 µL of the appropriate treatment was inserted in a hole in
the screen of each container. The tops of the micropipette tips were covered with parafilm to prevent
evaporation. The tips were weighed prior to dosing. After four hours,the micropipettes were removed
and reweighed, and the honey bees were fed 50%sucrose ad libitum for the remainder of the study.
STILLMEADOW,inc.

26312-23
Page 9 of 29
PROCEDURES(cont.)
Observations
Bee Groups 1-V were observed at approximately 4, 24, and 48 hours after dosing for mortality and
clinical signs of toxicity,particularly signs of intoxication(ataxia,lethargy,hypersensitivity,etc.). Any
dead bees were not removed from the paper cup until the end of the study and all bees were frozen and
disposed of at termination of study. Relative humidity and temperature were recorded at the first daily
observation time.
Test Validity
ty
The test is considered valid if mortality of the bees in the control group is less than 10%at the end of
the test and the 24 hour LD50 of the toxic standard falls within 0.10-0.35 gg/bee.
LDsn for Positive Control
The LD50 for dimethoate was determined by Rosiello, Essignmann and Wogan: Rapid and Accurate
Determination of the Median Lethal Dose and its Error with a Small Computer,Journal Toxic Environ
Health, 797-809, 1977 Computed on Microsoft Office 97 Visual Basic copyright 1997.
Statistical Analysis
Statistical comparisons at 4, 24, and 48 hours, between the control group and test group,could not be
performed as both groups' standard deviation was zero.
Evaluation of Results
If test group mortality did not exceed the controls and not more than 10%of control bees died during
the test,then the LD50 was considered greater than 100 µg a.i./bee.
Percent mortality was calculated at each observation point using the following formula:
(Number of dead bees/Beginning number of bees)x 100
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26312-23
Page 10 of 29
RESULTS AND DISCUSSION
Data Not Used
Data generated from 31 May 23 through 02 Jun 23 was not used due to a control failure. Test was
restarted on 06 Jun 23.
Protocol Deviation
• Due to a technician error, the positive control groups were dosed at a higher concentration;
however,an LD50 could still be calculated, and it was in range for validity requirements. This
deviation had no adverse effects on the outcome or integrity of the study.
Test Validity
The test was considered valid because control mortality(0%)was less than 10%and the 24 hour LD50
of the positive control was within 0.10-0.35 µg/bee,at 0.307 µg/bee.
Control,Test item and Toxic Standard Consumption
The amount of test item, sucrose solution or toxic standard consumed during the 4-hour dosing period
by the bees in each cup is presented in Table 2. The control and test groups consumed an average of
0.119 g and 0.117 g per replicate, respectively from the 100 µL offered of the appropriate substance
during the study. The toxic standard groups, at 0.025, 0.25 and 2.5 µg/bee, consumed an average of
0.085 g,0.086 g and 0.099 g per replicate,respectively,from 100 µL of toxic standard solution offered.
LD50 for Positive Control
The estimated LD5o for dimethoate at 24 hours was 0.307 µg/bee with 95% confidence levels
of 0.2293 -0.4507 µg/bee. The statistical analysis printout is included as Appendix D.
STILLMEADOW.Inc.

26312-23
Page 11 of 29
RESULTS AND DISCUSSION (cont.)
Clinical Signs of Toxicity, Mortality Observations and Evaluation
Clinical signs of toxicity and mortality observations are presented in Table 3. Statistical comparisons
at 4,24,and 48 hours,between the control group and test group,could not be performed as both groups'
standard deviation was zero.
At all observations, all bees in the control group and the test item group were NOA, meaning no
observed abnormalities. All of the surviving bees in the positive control group treated with 2.5 µg/bee
dimethoate seemed to be agitated at hour 4 and one bee in the positive control group treated with 0.25
µg/bee dimethoate was moribund at hour 4.
At 48 hours,mortality in the control and test groups was 0%(Table 1). Because the mortality of the
test group was the same as the control group mortality(0%),there is no difference and the LD50 of Lake
Guard Oxy is considered greater than 100 µg a.i./bee.
Table 1 -Mean Number of Dead Honey Bees and Percent Mortality Summary
Mean Dead per Cup/Total%Mortality
Group* Time Post-Dose
4 Hours 24 Hours 48 Hours
Control 0.0/0.0 0.0/0.0 0.0/0.0
Lake Guard Oxy 0.0/0.0 0.0/0.0 0.0/0.0
Dimethoate 0.025 µg/bee 0.0/0.0 0.4/2.0 0.4/2.0
Dimethoate 0.25 µg/bee 1.4/7.0 8.0/40.0 8.2/41.0
Dimethoate 2.5 µg/bee 9.8/49.0 19.6/98.0 20.0/100.0
Each group began with 100 honey bees on day 0.
STILLMEADOW,Inc.

26312-23
Page 12 of 29
CONCLUSION
This study was designed to assess the acute oral toxicity potential of the test item, Lake Guard Oxy,
when administered in the diet of the adult worker honey bee, Apis mellifera, at 100 µg active
ingredient(a.i.)/bee. The test item and toxic standard were administered orally in a 50%sucrose
solution to honey bees for four hours,with a control group fed only sucrose solution.
Bees were offered 100 µL of the test substance solution with a nominal concentration of 20 µg a.i./µL.
For a total concentration of 2000 µg a.i./100 AL,giving a nominal concentration of 100 µg a.i./bee in a
cup of 20 bees.
The estimated Median Lethal Dose(LD50) for dimethoate at 24 hours was 0.307 µg/bee, which falls
within the validity requirement of 0.10-0.35 µg/bee.
At 48 hours, mortality for the control group, test group, and positive control groups(0.025, 0.25 and
2.5 µg/bee dimethoate), was 0%, 0%, 2%,41%and 100%,respectively. The control and test groups
consumed an average of 0.119 g and 0.117 g per replicate,respectively,from the 100 µL offered of the
appropriate substance during the study. The validity requirement of no more than 10%control mortality
was met with a control mortality of 0%. The test group mortality at 0%did not exceed control mortality,
therefore,the difference was not significant and the LD50 of Lake Guard Oxy is considered greater than
the nominal concentration of 100 µg a.i./bee.
/ //‘
Cole Younger,PhD Date
Study Director
Entomologist,STILLMEADOW,Inc.
STUDY PERSONNEL
Technical Staff Technical Writers
Stephen Balestrier,BS Alexis Wiltshire, BS
Mariana Cortez,AAS Sharon Barrow,BS
Hugo Martinez
STILLMEADOW,Inc.

1
26312-23
Page 13 of 29
Table 2- Dose Consumption
Honey Bee Acute Oral Toxicity Limit Test
Test Substance: Lake Guard Oxy
Group 1-Vehicle Control Group IV-0.25 µg/bee Dimethoate
Cup Day 0 After 4 Hours Amount Cup Day 0 After 4 Hours Amount
Number Given(g) Remaining($) Consumed(ej Number Given(s) Remaining(g) Consumed(0
1 1.0999 0.9909 0.1090 1 1.0923 0.9646 0.1277
2 1.0889 0.9867 0.1022 2 1.0916 1.0658 0.0258
3 1.1000 0.9710 0.1290 3 1.0848 0.9465 0.1383
4 1.0992 0.9727 0.1265 4 1.1013 1.0949 0.0064
5 1.0777 0.9506 0.1271 5 1.0836 0.9513 0.1323
Mean 1.0931 0.9744 0.1188 Mean 1.0907 1.0046 0.0861
SD 0.0098 0.0158 0.0123 SD 0.0071 0.0702 0.0644
SE 0.0044 0.0071 0.0055 SE 0.0032 0.0314 0.0288
Group II- Lake Guard Oxy Group V-2.5 µg/bee Dimethoate
1 Cup Day 0 After 4 Hours Amount Cup Day 0 After 4 Hours Anwunt
Number Given(g) Remaining(g) Consumed(g) Number Given( Remaining(g) Consumed 0)
1 1.0383 0.923 0.1153 1 1.0880 0.9773 0.1107
2 1.0489 0.9216 0.1273 2 1.0853 0.9659 0.1194
3 1.0292 0.9013 0.1279 3 1.0492 0.9418 0.1074
4 1.0731 0.9525 0.1206 4 1.0836 0.9969 0.0867
5 1.0940 1.0012 0.0928 5 1.1094 1.0408 0.0686
Mean 1.0567 0.9399 0.1168 Mean 1.0831 0.9845 0.0986
SD 0.0265 0.0388 0.0144 SD 0.0216 0.0372 0.0206
SE 0.0119 0.0174 0.0064 SE 0.0097 0.0167 0.0092
Group III-0.025 µg/bee Dimethoate
Cup Day 0 After 4 Hours Amount
Number Given(g) Remaining( Consumed
1 1.1061 0.9640 0.1421
2 1.1184 0.9855 0.1329
3 1.1355 1.0065 0.1290
4 1.1183 1.1113 0.0070
5 1.0762 1.0601 0.0161
Mean 1.1109 1.0255 0.0854
SD 0.0220 0.0598 0.0677
SE 0.0099 0.0267 0.0303
SD.standard deviation;SE,standard error
STILLMEADOW.Inc.

1
I

1
26312-23
Page 14 of 29
Table 3- Mortality, Observations and Percent Mortality
Honey Bee Acute Oral Toxicity Limit Test
Test Substance: Lake Guard Oxy
Observations and Percent Mortality
Group I-Vehicle Control
Hours Number of Dead Bees Mean Dead Total Percent
Post-dose Cu, 1 Cu. 2 Cu, 3 Cu. 4 C • 5 .er Cu, Dead Mortali Observations
4 0 0 0 0 0 0.0 0 0.0% NOA
24 0 0 0 0 0 0.0 0 0.0% NOA
48 0 0 0 0 0 0.0 0 0.0% NOA
Mortality(%) 0.0% 0.0% 0.0% 0.0% 0.0% Total 0 0.0%
Group II-Lake Guard Oxy
Hours Number of Dead Bees Mean Dead Total Percent
Post-dose Cup 1 Cup 2 Cup 3 Cup 4 Cup 5 per Cup Dead Mortality Observations
4 0 0 0 0 0 0.0 0 0.0% NOA
24 0 0 0 0 0 0.0 0 0.0% NOA
48 0 0 0 0 0 0.0 0 0.0% NOA
Mortality(%) 0.0% 0.0% 0.0% 0.0% 0.0% Total 0 0.0%
Group Ill-0.025 pg/bee Dimethoate
Hours Number of Dead Bees Mean Dead Total Percent
Post-dose Cup I Cup 2 Cup 3 Cup 4 Cup 5 per Cup Dead Mortality Observations
4 0 0 0 0 0 0.0 0 0.0% NOA
24 0 1 I 0 0 0.4 2 2.0% Rest NOA
48 0 1 I 0 0 0.4 2 2.0% Rest NOA
Mortality(%) 0.0% 1.0% 1.0% 0.0% 0.0% Total 2 2.0%
Group IV-0.25 pg/bee Dimethoate
Hours Number of Dead Bees Mean Dead Total Percent
Post-dose Cu, I Cu. 2 Cu. 3 Cu. 4 Cu, 5 .er Cu. Dead Mortali Observations
4 6 0 0 0 I 1.4 7 7.0% I moribound cup
1,Rest NOA
24 16 0 19 0 5 8.0 40 40.0% Rest NOA
48 16 0 20 0 5 8.2 41 41.0% Rest NOA
Mortality(%) 16.0% 0.0% 20.0% 0.0°/e 5.0% Total 41 41.0%
Group V-2.5 pg/bee Dimethoate
Hours Number of Dead Bees Mean Dead Total Percent
Post-dose Cup I Cup 2 Cup 3 Cup 4 Cup 5 ,per Cup Dead Mortality Observations
a
4 14 6 I 20 8 9.8 49 49.0%
All remaining bees
seem agitated
24 20 20 20 20 18 19.6 98 98.0% Rest NOA
48 20 20 20 20 20 20.0 100 100.0% NA
Mortality(%) 20.0% 20.0% 20.0% 20.0% 20.0% Total 100 100.0%
NOA-No observable abnormalities;NA-Not Applicable
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26312-23
Page 15 of 29
Appendix A- Signed Protocol
STILLMEADOW
INCORPORATED
PROTOCOL FOR STUDY 26312-23
Study Title: Honey Bee,Apis mellffera,Acute Oral Toxicity Limit Test
(OECD 213)
Test Item: Lake Guard Oxy
Test Facility: STILLMEADOW,Inc.
12852 Park One Drive
Sugar Land,TX 77478
7/
Approved:
Cole Younger, Date
Study Director
STILLMEADOW,Inc.
Approved: /�Y G tn�- 0 8 u 01
Management Date
STILLL�,�MB��AD� OW,Inc.dev
Reviewed: % ;; 4te 0 a a`i
Kristina Rodrigue,RQ GLP Date
Quality Assurance Dr or
STILLMEADOW,Inc.
Sponsor: Bluet3reen Water Technologies
3101 South Hills Village Ste LL200#452
Pittsburgh,PA 15241
866-269-2828
jessica.frostRblueRreenwateItech.corn
1LcP.`-- 51o//40ya
Approved:
Jessica Frost Date
Scientific Director
12852 Park One Drive• Sugar Land,Texas 77478 in 281 240-8828 ■ Fax [phone]
www.stillmeadow.com
STILLMEADOW.Inc.

26312-23
Page 16 of 29
Appendix A-Signed Protocol (cont.)
Protocol for Study 26312-23
Page 2 of 6
PROTOCOL FOR STUDY 26312-23
A. GENERAL
1. Study Title; Honey Bee,Apis mellifera,Acute Oral Toxicity Limit Test
2. Purpose: To assess the acute oral toxicity potential of the test item when
administered to adult worker honey bees.
3. Methods Guidelines: The study will be conducted according to OECD Guideline 213.
4. Regulatory Compliance: This study will be conducted in compliance with Good Laboratory
Practice Standards of EPA FIFRA,40 CFR Part 160 and OECD
ENV/MC/CHEM(98)17.
The dose mixtures will not be verified according to the
requirements specified in 40 CFR 160.113(a) and
ENV/MC/CHEM(98)17 6.2(5).
In the event of a regulatory inspection,Regulatory Inspectors will
be provided with all study documentation requested. The Sponsor
will be notified of the inspection of their study.
All methods can be found in STILLMEADOW, Inc. Standard
Operating Procedures(SOPs).
5. Quality Assurance: The Quality Assurance Unit(QAU)will review the protocol. The
study information will be entered into the Master Schedule. In-
progress inspection(s)will be performed to ensure the integrity of
the study. Any deviations from SOPs, the Protocol or Good
Laboratory Practice Standards will be immediately reported to the
Study Director and Management. The raw data and report will be
audited,and a statement prepared and signed which will specify
the dates that the inspections were made,and findings reported to
Management and the Study Director.
6. Test Item; Lake Guard Oxy. Test item identification should include the
name,batch number and purity. The Sponsor should also provide
information regarding safety, stability, storage conditions and
disposal. The Sponsor assumes responsibility for purity,stability,
identity,synthesis methods and location of documentation.
7. Control Item: Dimethoate CAS#60-51-5.
8. Pr chedule; Proposed Experimental Start Date: 22 May 23
Proposed Experimental End Date: 25 May 23
Study duration: at least 48 hours, and may be extended to a
maximum of 96 hours after dosing.
9. Study Director: Cole Younger,PhD
(Rev:05May23) STILLMEADOW,Inc.
STILLMEADOW,Inc.

26312-23
Page 17 of 29
Appendix A-Signed Protocol(cont.)
Protocol for Study 26312-23
Page 3 of 6
A. GENERAL(cont.)
10. Experimental Summary__ The test item, and serial dilution concentrations of the toxic
standard will be administered orally to honey bees in a 50%
sucrose solution for four hours and the amount consumed will be
recorded. A group of bees will be fed the 50%sucrose solution
alone for comparison. The bees will be observed for mortality and
clinical signs of toxicity at --4, 24 and 48 hours after dosing.
Observations may be extended to 96 hours after dosing.
11. Protocol Amendments: Any alteration in the Protocol will he justified, approved by the
Study Director and recorded in writing.
12. 5sonsor Audits: The Sponsor may send an authorized representative to inspect the
test system and/or data on the STILLMEADOW, Inc. premises
during normal working hours.
B. EXPERIMENTAL DESIGN
1. Insects
a. Species/Source: Italian honey bee,Apis mellra,worker;STILLMEADOW,Inc.
bee colony, disease and pest-free with no previous pesticide
exposure.
b. Justification of Species: The honey bee is the species required in the regulatory guidelines
for this study.
c. Quantity: S00 bees; 100 bees per group in five replicates of 20 bees each.
Five groups:Control,Test and three toxic standard levels.
d. Age at Dosing: Young adult worker bees similar in age.
c. Identification: Replicates will be labeled according to treatment.
f. Acclimation and
Health Status: No acclimation is necessary. Normal appearance and behavior
will be factors used to select healthy bees from disease-free
colonies for testing.
2. Insect Husbandry
a. Exterior housing: Standard commercial honey bee hive
b. Indoor Chambers: Disposable cardboard containers with a screened lid
(Rev:05May23) STILLMEADOW,Inc.
STILLMEADOW,Inc.

26312-23
Page 18 of 29
Appendix A- Signed Protocol(cont.)
Protocol for Study 26312-23
Page 4 of 6
B. EXPERIMENTAL DESIGN(cont.)
llusbandr cont.2. Insect y(cont.)
c. Food: 50:50 w/v sucrose:dechlorinated (DC)water solution; available
ad libitum after test dose is removed.
d. Environment: Incubator temperature:33*5°C.
Target relative humidity:50-70%
Honey bees will be kept in dark except when dosing or making
observations.
e. Handling: Only as much as is necessary to conform to test procedures;
shielded from excessive activity or other disturbance during
holding and testing.
3. Group Allocation and
Randomization: Honey bees will be immobilized once using CO2,randomly taken
from the collection container and placed in one of five groups.
4. Preparation of Test Bees: Bees will be fasted for approximately two hours before dosing.
Any bees found moribund during this time will be rejected and
replaced by healthy bees fasted concurrently before starting test.
5. Dose Calculation and
Preparation: Dosing calculation will be based on 100%test item concentration.
To prepare a 2000 pg a.iJ100 µL solution,200 mg of test item
will be dissolved into 10 mL of 50%sucrose solution. According
to the following calculations:
2000 µg/100 pL* 10(convert to mL)-20000 µg/mL
20000 pg/mL/I,000,000(convert to g)=0.02 g/mL
Solution will be prepared in 10 mL;therefore 0.020 g x 10 mI,=
0.20 g(200 mg)for every 10 mL of 50%sucrose solution prepared
Per Sponsor information,the solution will be allowed to sit for one
hour(±15 minutes)before dosing to allow the active substance to
be released from the coating. Solution will be shaken prior to use.
6. Dose Level: A limit test of 100 µg a.i/bee will be conducted with the test item
being administered for four hours as a single dose level to five
replicates of 20 bees each. The bees will be observed for mortality
and clinical signs of toxicity at-4,24 and 48 hours. The dosing
solution will be prepared as a 2000 µg a.i./100 µL solution,based
on the Sponsor-provided nominal claim of active ingredient
concentration. A 100 µL aliquot of the 2000 µg a.i.solution is the
equivalent of offering 100 pg a.i.per bee.
7. Vehicle Selection: 50%sucrose will be used as the vehicle.
(Rev:05May23) STILI.MF,AROW,Inc.
" l ll t.3lt: DONN. Inc.

26312-23
Page 19 of 29
Appendix A- Signed Protocol(cont.)
Protocol for Study 26312-23
Page5of6
B. EXPERIMENTAL DESIGN(cont.)
8. Test Item
Administration: On Day 0,each container of bees will be randomly assigned to
either the test item, toxic standard or control sucrose solution
group. For dosing,a micropipette tip containing 100 µL of the
appropriate solution will be inserted in a hole in the screen for
each container. The top of the micropipette tips will be covered
with parafilm to prevent evaporation. The tips will be weighed
prior to dosing and at four hours to determine the amount
consumed. After four hours,honey bees will be fed 50%sucrose
and DC water ad libitum.
9. Untreated Control; An untreated control group will be conducted concurrently. The
untreated sucrose solution will be administered to five replicates
of 20 bees each as a single dose level equivalent to the amount
given to the test group.
10. Toxic Standard: A toxic standard (positive control) will be conducted
concurrently. Three dose levels(0.01,0.1,and 1.0 µg/bee)of the
toxic standard,dimethoate,will be administered to five replicates
of at least 20 bees. Toxic standard groups will receive the
dimethoate in the same manner as test and controls.
11. Qtiservations; All bees will be observed at—4,24 and 48 hours after dosing for
mortality and clinical signs of toxicity, particularly signs of
intoxication(ataxia,lethargy,hypersensitivity,etc). If mortality
in the test group is greater than 10%at 48 hours and the untreated
control mortality is<10%,observations will be extended to 72 and
96 hours. Any dead bees will not be removed until the end of the
study and all bees will be frozen and disposed of at the end of the
study. Relative humidity and temperature will be recorded at the
first daily observation time.
12. Test Validity: For the test to be considered valid,control mortality cannot exceed
10% at the end of the test and the 24 hour LDso of the toxic
standard falls within 0.10-0.35 µg a.i./bee.
13. evaluation of Results: If mortality does not significantly exceed the controls and not
more than 10%of control bees die during the test,the LDso will
be considered greater than 100 µg a.i./bee.
14. Test Item
Accountability: A comprehensive inventory of test item received and used will be
kept. The test item container(s)will be weighed when received at
this facility,and a record of all test item use will be maintained.
Test item and test item dosing solutions will be stored in the
original containers or equivalent, or in glass containers with
Teflon-lined caps.
(Rev:05May23) STILLMEADOW,Inc.
STILLMEADOW,Inc.

26312-23
Page 20 of 29
Appendix A- Signed Protocol(cont.)
Protocol for Study 26312-23
Page 6 of 6
B. EXPERIMENTAL DESIGN(cont.)
L 5. Disposal of Unused
Test Item: Unused test item will be disposed of at Sponsors expense after
termination of the study.
16. Safety Precautions: General safety precautions required by laboratory SOPS will be
followed.The Sponsor will supply basic toxicity data on the test item
to be used. However,since the toxicity of test items is often not well
characterized,this laboratory will be conservative in setting safety
procedures. The Sponsor or Sponsors Representative shall be
notified of any exposures requiring a physician's examination or care.
C. DATA MANAGEMENT
1. Records: The following records will be maintained during the study and
archived at STILLMEADOW,Inc.upon study termination.
a. Protocol and Protocol Amendments(if any)
b. Final report and amendments(if any)
c. Study correspondence
d. Bee procurement data
c. Test item receipt,identification as supplied by the Sponsor,
preparation,administration and disposition
f. Test insect information: number, species, source and hive
number.
g. Daily clinical signs and mortality,if any
h. Other pertinent data
2. Data Storage: All raw data, original protocol, original final report, any
amendment(s),and a retained test item sample will be archived at
STILLMEADOW,Inc.for a period of 15 years.
3. Data Reporting: The final report will include all data as described in the Good
Laboratory Practice Standards,Including:
a. Statement from the Quality Assurance Unit
b. Signature of the Study Director
c. A GLP Compliance Statement signed by the Study Director
d. Names of scientific personnel involved in the study
c. Dates of study initiation and termination
f. Identification, label information, description and storage of
the test item,and identification of the vehicle used
g. All pertinent honey bee information and observation methods
h. Description of the test procedures
i. Daily observations for mortality and clinical signs of toxicity
j. A copy of this Protocol
k. Any deviations and the impact,if any,on the study
4. Report Generation; A final report will be generated after completion of the laboratory
portion of the study.
(Rev:05May23) STILLMEADOW,Inc.
STILLMEADOW.Inc.

26312-23
Page 21 of 29
Appendix B-Sponsor Provided Product Label
UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
WASHINGTON. D.C. 20460
OFFL.E'FCHEi ,AL SAFE TY
Atal POW/nirr PREVENT IOU
March 6.2023
Brian Hogan
Agent for Bluegreen US Water Technologies.Inc.
c o'MI:Consulting LLC
5807 Churchill Way
Medina.0 44256
Subject: Labeling Notification per Pesticide Registration Notice(PRN)98-10—Delete statement
related to shallow water
Product Name:Lake Guard Oxy
EPA Registration Number:93647-2
Application Date:01 30'2023
OPP Case Number:00428733
Dear Mr.Hogan:
The U.S.Environmental Protection Agency(EPA)is in receipt of your application for notification under
Pesticide Registration Notice(PRY)98-10 for the above referenced product.The Biopesticides and
Pollution Prevention Division(BPPD)has conducted a review of this request for its applicability under
PRN 98-10 and finds that the action requested falls within the scope of PRN 98-10.
The labeling submitted with this application has been stamped-Notification"and will be placed in our
records.You must submit one(1)copy of the final printed labeling with the modifications.
At this time.the t.S.Environmental Protection Agency(EPA)is not requiring a change to the
restricted-entry interval(REI)of 0 hours(for non-spraying methods)and 1 hour(for spraying methods 1
as it appears on your labeling in the Agricultural Use Requirements Box.The EPA is currently
developing guidance and criteria for when it will allow a REI of less than 4 hours and might require a
change to the REI for this product in the finure.
Should you wish to add retain a reference to your company's website on your label.then please be
aware that the website becomes labeling under the Federal Insecticide.Fungicide.and Rodenticide Act
(FIFRAI and is subject to review by the EPA.If the website is false or misleading.the product will be
considered to be misbranded and sale or distribution of the product is unlawful under FIFRA section
121a)(11(E).40 CFR si 156.100)(5)lists examples of statements the EPA may consider false or
misleading.In addition,regardless of whether a website is referenced on your product's label.claims
made on the website may not substantially differ from those claims approved through the registration
process.Therefore.should the EPA find or if it is brought to our attention that a website contains false
STILLMEADOW.Inc.

26312-23
Page 22 of 29
Appendix B-Sponsor Provided Product Label (cont.)
Page 2 of 2
EPA Reg.No.93647-2
OPP Case No.00428733
or misleading statements or claims substantially differing from the EPA-approved registration,the
website will be referred to the EPA's Office of Enforcement and Compliance Assurance.
If you have any questions.please contact Susannah Powell via email at powell.susannah:aepa.gov.
Sincerely.
}
James Parker,Team Leader
Biochemical Pesticides Branch
Biopesticides and Pollution
Prevention Division(75l 1P)
Office of Pesticide Programs
STILLMEADOW.Inc.

26312-23
Page 23 of 29
Appendix B- Sponsor Provided Product Label (cont.)
NUTIHK.V1 IIM
„gym;; LAKE GUARD Oxy
Algaecide/Cyanobacteriocide
[Large Granu es][Small Granules][Dust]
ACTIVE INGREDIENT
Sodium percarbonate ......... 83.3%
OTHER INGREDIENTS 16.7%
TOTAL 100.0% ceroiea to
NSF+ANS6'CAN 60
[UN 3378}
[CLASS 5 1] MUL 33 mg/l
KEEP OUT OF REACH OF CHILDREN
DANGER/PELIGRO
Si ust*d no entiende la eti ueta.busque a al uien para quit se la explique a usted en detail*.
9 8
Of you do not understand the Zabel.find someone to explain it to you in detail.)
FIRST AID
If in eyes •Hold eye open and rinse slowly and gently with water for 15-20 minutes Remove
contact lenses.if present,after the first 5 minutes,then continue rinsing.
•Call a poison control center or doctor for treatment advice.
If swallowed • Call a poison control center or doctor immediately for treatment advice
• Have person sip a glass of water if able to swallow
• Do not induce vomiting unless told to by a poison control center or doctor
• Do not give anything to an unconscious person
If on stun •Take off contaminated clothing
• Rinse skin immediately wrth plenty of water for 15-20 minutes.
• Call a poison control center or doctor for treatment advice
If inhaled • Move person to fresh air
•If person is not breathing,call 911 or an ambulance then give artificial respiration.
preferably mouth-to-mouth if possible.
• Call a poison control center or doctor for further treatment advice.
Have the product container or label with you when calling a poison control center,doctor,or going for
treatment For non-emergency mformabon concerning this product.call the National Pesticides
Information Center iNPIC)at 1-800-858-7378 iNPIC Web site.www.nplc.orst.edu) For emergent es.
call the poison control center 1-800-222-1222
NOTE TO PHYSICIAN Probable mucosal damage may contraindicate the use of gastric lavage
EPA Reg No 93647-2
EPA Est No XXXXX-XX-X
Net Contents 2,5.10,20,25,50,500,1.000.2.000 2204 lbs
Batch Code
Page 1 of 5
STILLMEADOW,Inc.

26312-23
Page 24 of 29
Appendix B- Sponsor Provided Product Label (cont.)
PRECAUTIONARY STATEMENTS
Hazards to Humans and Domestic Animals
DANGER`PELIGRO
Corrosive.Causes irreversible eye damage and causes skin burns.May be fatal if swallowed Harmful if
absorbed through skin or inhaled.Do not get in eyes,on skin,or on clothing.Avoid breathing dust.Wear
protective eyewear.such as goggles face shield,or safety glasses.Wash thoroughly with soap and
water after handling and before eating,drinking,chewing gum. using tobacco or using the toilet.Remove
and wash contaminated clothing before reuse.
Environmental Hazards
This pesticide is toxic to birds Do not apply this product or allow it to drift to blooming crops or weeds
while pollinating insects are actively visiting the area.
For container sizes 50 lbs.or greater: Do not discharge effluent containing this product into lakes, •
streams,ponds estuaries oceans or other waters unless in accordance with the requirements of a
National Pollutant Discharge Elimination System(NPDES)permit and the permitting authority has been
notified in writing pnor to discharge.Do not discharge effluent containing this product into sewer systems
without previously notifying the local sewage treatment plant authority For guidance contact your State
Water Board or Regional Office of U.S.EPA.For additional information,refer to the product
Material Safety Data Sheet.
Physical and Chemical Hazards
Oxidizing agent Never use with other pesticides.cleaners or oxidizing agents
Personal Protective Equipment
Corrosive.Mixers.loaders,applicators.and other handlers must wear the following.
• Long-sleeved shirt and long pants
• Chemical resistant gloves
• Protective eyewear(googles or face shield)
• Shoes plus socks
Follow manufacturer's instructions for cleaning maintaining PPE If no such instructions for washables
exist.use detergent and hot water. Keep and wash PPE separately from other laundry
USER SAFETY RECOMMENDATIONS
Users should remove clothing/PPE immediately if pesticide gets inside.Then wash thoroughly and put
on clean clothing.
Users should remove PPE immediately after handling this product.Wash the outside of gloves before
removing As soon as possible,wash thoroughly and change into clean clothing.
DIRECTIONS FOR USE
It is a violation of Federal Law to use this product in a manner inconsistent with its labeling.
GENERAL APPLICATION RESTRICTIONS
Do not apply this product in a way that will contact workers or other persons either directly or through
drift.Only protected handlers may be in the area during application For any requirements specific to your
State or Tribe,consult the State or Tribal agency responsible for pesticide regulation
This product is not intended as treatment against any public health organism for any use on this label
Do not apply directly to treated,finished drinking water reservoirs or drinking water receptacles when the
water is intended for human consumption
SPRAY DRIFT MANAGEMENT
Page 2of5
STILLMEADOW.Inc.

26312-23
Page 25 of 29
Appendix B-Sponsor Provided Product Label (cont.)
A variety of factors including weather conditions(e.g.,wind direction,wind speed.temperature,and
relative humidity l and the method of application(e g..ground or aerial)can influence pesticide drift The
applicator must evaluate all factors and make appropriate adjustments when applying this product.
Wind Speed
Do not apply at wind speeds greater than 15 mph Only apply this product if the wind direction favors on-
target deposition(approximately 3 to 10 mph).and there are no sensitive areas within 250 feet downwind
•
Temperature Inversions
If applying at wind speeds less than 3 mph.the applicator must determine if a)conditions of temperature
inversion exist.or b)stable atmospheric conditions exist at or below nozzle height. Do not make
applications into areas of temperature inversions or stable atmospheric conditions
Other State and Local Requirements
Applicators must follow all state and local pesticide drift requirements regarding application of hydrogen
peroxide compounds Where states have more stringent regulations.they must be observed
Equipment
All aerial and ground application equipment must be properly maintained and calibrated using appropriate
carriers or surrogates
For aerial applications:
The boom length must not exceed 75%of the wingspan or 90%of the rotor blade diameter.Release
granules at the lowest height consistent with efficacy and flight safety When applications are made with
a crosswind,the swath must be displaced downwind The applicator must compensate for this
displacement at the up and downwind edge of the application area by adjusting the path of the aircraft
upwind
GENERAL INFORMATION
Water bodies,such as impounded water raw-water for drinking.lakes ponds.reservoirs,waste water,
irrigation.drainage.aquaculture. conveyance ditches pipes,canals.laterals,estuaries.bayous.lagoons,
and brackish and salt.sea and ocean water contaminated with algaercyanobactena can be treated with
Lake Guard'Oxy
Do not apply when wind speed favors dnft beyond the area intended for treatment
•
ciagaaat.—Do not use the product to treat ornamental fish(i.e.,fish that grow in home aquariums or
outdoor tanks for aesthetic purposes).
Water hardness.temperature of the water.type and cell density of algae:'cyanobactena to be controlled.
and the amount of water flow are to be considered in using Lake Guard*Oxy to control
algaercyanobacteria Treated water resource should be monitored systematically for the presence of
harmful algae:cyanobactena using adequate apparatuses.Begin treatment as soon algalicyanobacterial
cell numbers reach 5,000-20,000 cells/mL(and below 10 pg chlorophyll-after).
If treatment is delayed until algalicyanobacterial cell numbers exceed 20.000 celtlmL(or equivalently.
above 10 pg chlorophyll-a!liter), an increase in the quantities of Lake Guard*Oxy will be required.as well
as in treatment frequency Use caution when treating heavy blooms.as oxygen loss from mass
decomposition of dead algaercyanobactena can cause fish suffocation See application rates section for
treating heaving blooms.
Always apply the granules off wind and let the wind and currents to carry them where
algae'cyanobactera cell masses are concentrated.It is best to treat algae on a sunny day at morning
hours If there is some doubt about the amount of Lake Guard*Oxy to apply.it is best 10 start with the
lower application rate The quick and easy treatment application(minutes for applying a hundred lb(,and
Page 3 of 5
STILLMEADOW.Inc.

26312-23
Page 26 of 29
Appendix B- Sponsor Provided Product Label (cont.)
the visual results that would be apparent within 48-72 hours,enable the applicator to make a quick
adjustment on whether to increase the dose rate by 50-100%during the next treatment application or
scale it back and use even less product under similar conditions.Since all water bodies are unique
aquatic ecosystems,and therefore,differ from each other in treatment response and longevity,the Lake
Guards Oxy application protocol offers the applicator a scalable dose rate that saves money and reduces
the chemical load introduced to the environment For first time users.or when in doubt,applicators should
start with 0 5-5 Ib/acre and adjust the dose rates(increase or decrease)by increments of 25-50%.and
over time can fine-tune the treatment application for each waterbody.
APPLICATION RATES
The best method by which to apply Lake Guards Oxy granules to water is by broadcasting(dusting)it
over a well-defined contamination zone,at early bloom stages,when harmful algallcyanobacteria
numbers are at 5,000 to 20,000 cells(mL Iand below 10 pg chlorophyll-Miter).
Determination of surface area to be treated
Determine the size of the infested area as follows*.(1)in small infested reservoirs.under than 250 acres
obtain surface area by measunng of regular shaped ponds or mapping of irregular ponds or by reference
to previously recorded engineering data or maps (2)In water bodies larger than 250 acres tor smaller
ponds with a defined contaminated zones outline the infested area by a combination of the following
instruments:microscopical count,pigment extraction.toxin evaluation,probes that detect specific
pigments that are known to serve as a correlated proxy for algae/cyanobacteria biomass.satellite
imaging,etc.NOTE.evaluation of the state of the infestation should be done by professional personnel.
Determination of the aeelication rate
For control of harmful algae:cyanobacteria infestation it is essential to begin Lake Guards Oxy treatment
when harmful algae!!cyanobactena cell numbers are in the range of 5.000-20,000 cell/mL(or below 10 pg
chlorophyll-Miter) Apply 0 5 5 lbs.`acre Lake Guards Oxy at these algal/cyanobacterial cell-densities
Always start with the lower rate.At higher infestation rates.when cyanobactenal cell density is between
20.000-100,000 cells/mt.(or between 10-50 pg chlorophyll-a//liter)use 5-301bs.'acre Lake Guards Oxy.
If treatment is delayed until algal/cyanobacterial cell numbers exceed 100,000 cellimL(or equivalently,
above 50 pg chlorophyll-Miter),an increase in the quantities of the Lake Guards Oxy will be required,as
well as in treatment frequency Therefore,in heavy blooms,when cyanobacterial scum or aggregates are
visible to the naked eye (more than 100.000 cells/mL of algaei`cyanobacteria or over 50 ug chlorophyll-
Miter),treat with doses between 30-98 lbs acre.If doses exceed 98 lbs/acre_treat no more than one-
half of the water area in a single application Maximum single application rate allowed should not exceed
294 lbs/acre of the Lake Guards Oxy NOTE when cyanobacterial aggregated could be seen with the
naked eye,the cyanobacterial cell density in the water is estimated to exceed 100.000 cells per ml
When a single application dose is below 30 lbs'acre,minimum retreatment interval is 12 hours When a
single application dose is between 30-98 lbs./acre.minimum retreatment interval is 24 hours When a
single application dose exceeds 98 lbs./acre the minimum retreatment interval is 48 hours.
APPLICATION METHOD
Apply Lake Guard'Oxy using equipment designed for granular dusting.Dusting can be done manually by
hand or by a boat or airplane,depending on the area of the zone and its proximity to reservoir's bank
When a small duster is mounted on a properly equipped boat. application can be broadcast directly on
the water surface at the edge of the infested zone.Note that the direction of the wind is an important
factor-always dust off-wind.Do not use this method unless completely familiar with this type of
application.
Page 4 of 5
STILLMEA DOW.Inc.

26312-23
Page 27 of 29
Appendix B- Sponsor Provided Product Label (cont.)
STORAGE AND DISPOSAL
Do not contaminate water.food,or feed by storage or disposal
PESTICIDE STORAGE.Keep pesticide m original container Do not use in food or drink containers
PESTICIDE DISPOSAL Pesticide wastes may be hazardous Improper disposal of excess pesticide.
spray mixture or nnsate is a violation of Federal Law If these wastes cannot be disposed of by use
according to label instructions.contact your State Pesticide or Environmental Control Agency,or the
Hazardous Waste Representative at the nearest EPA Regional Office for guidance
CONTAINER HANDLING Nonrefillable container Do not reuse or refill this container Completely
empty bag into application equipment,then offer for recycling if available or dispose of empty bag in a
sanitary landfill or by incineration
CONDITIONS OF SALE AND LIMITATIONS OF WARRANTY AND LIABILITY
Read the entire directions for use.conditions of warranties and limitations of liability before using this
product If terms are not acceptable,return the unopened product container at once.By using this
product.user or buyer accepts the following Conditions.Disclaimer of Warranties and Limitations of
Liability.CONDITIONS:The directions for use of this product are believed to be adequate and must be
followed carefully However.a is impossible to eliminate all nsks associated with the use of this product.
Ineffectiveness or other unintended consequences may result because of such factors as weather
conditions,presence of other materials.or the manner of use or application.all of which are beyond the
control of BlueGreen Water Technologies All such risks shall be assumed by the user or buyer
DISCLAIMER OF WARRANTIES:To the extent consistent with applicable law.BlueGreen Water
Technologies makes no other warranties.express or implied,of merchantability or of fitness for a
particular purpose or otherwise,that extend beyond the statements made on this label No agent of
BlueGreen Water Technologies is authorized to make any warranties beyond those contained herein or to
modify the warranties contained herein.To the extent consistent with applicable law.BlueGreen Water
Technologies disclaims any liability whatsoever for special,incidental or consequential damages resulting
from the use or handhng of this product.LIMITATIONS OF LIABILITY:To the extent consistent with
applicable law,the exclusive remedy of the user or buyer for any and all losses,injuries or damages
resulting from the use or handling of this product,whether in contract.warranty.tort.negligence.strut
liability or otherwise,shall not exceed the purchase price paid or at BlueGreen Water Technologies'
election,the replacement of product.
Property rights are protected under the patent legislation of-USA Patent No 10,729,138 and USA Patent
No. 10.092.005:Indian Patent no.201617001647:Mexican Patent No.MX/a/2016/000199:South
African Patent No 2016100478:the Russian Federation Patent No.2687929.
Manufactured for:
BlueGreen US Water Technologies,Inc.
Address:301 South Hills Village Ste LL200#452 Pittsburgh,PA 15241
[e-mail]
[e-mail]
Page_5of5
STILLMEADOW.Inc.

r
26312-23
Page 28 of 29
Appendix C- Positive Control Certificate of Analysis
660 Tower Lane.P 0.Box 599•Wen Chester.PA 19331.0590
1•000-452.9994•I.610.6921026•Fa I.610-092.8719
latxt£ccra•1r099'1aas
CERTIFICATE OF ANALYSIS
Dimethoate
CATALOG NUMBER N-11758-100MG
LOT NUMBER 13914600
DATE CERTIFIED 03/07/22
EXPIRATION DATE 03/31/25
CAS NUMBER 60-51-5
MOLECULAR FORMULA C5H12NO3PS2
MOLECULAR WEIGHT 229 27
STORAGE Refrigerator storage(2-8°C)
HANDLING See Safety Data Sheet
INTENDED USE For laboratory use only.
Analytical Test Value
FT-IR SPECTROSCOPY CONFORMS TO STRUCTURE
%PURITY(GCGFID) 9A 0
Chem Service,Inc.guarantees the purity to be I!-0.5%deviation prior to the expiration date
shown cm the label and exclusive of any customer contamination.
•
•
•
Ccr6cd 8y. C l( (pP\) y iv? •
KI
K.r tu,a Jones tJ�
`r -m Chem Service,Inc Is accredited
COA Form �•. _� . :+ to 50AEC 170252017,
Revision 4.0(052023) .c c.w a a o 7...•.r c rr SO 17034 2016 and
certified to ISO 90012015
Print Date: 05/23/23
F.i.:r.
STILLMEADOW.Inc.

26312-23
Page 29 of 29
Appendix D- LDv,Analysis for Positive Control
24 Hours
LDH,Analysis•
Honey Bee,Apis mellifera,Acute Oral Toxicity Definitive Test
Study Number: 26312-23
Toxic Standard Dimethoate
Dose Level Number Number
pg/bee Dead Treated Mortality
2.5 98 100 98%
0.25 40 100 40%
0.025 2 100 2%
LDt 0.023 µg/bee
LDS 0.050 pg/bee
LDio 0.074 pg/bee
LD,s 0.102 pg/bee
95%lower confidence limit: 0.2293 pg/bee
I.11sa 0.307 pg/bee
95%upper confidence limit: 0.4507 pg/bee
LDu 0.9228 µg/bec
LD90 I.2739 pg/bee
LD95 1.8983 pg/bee
LD99 4.0373 pg/bee
Slope function(s)=3.02 with 95%confidence limits of 2.969 to 3.088.
Variance of Slope-0.9
Calculated X2=0.115 with 1 degrees of freedom.
Values for P=0.5 T= 12.7 X2" 3.84
•Rosiello,Essignmatm and Wogan: Rapid and Accurate Determination of the Median Lethal
Dose and its Error with a Small Computer,Journal Toxic Environ Health,797-809, 1977
Computed on Microsoft Office 97 Visual Basic copyright 1997
o�r, / 42 3 _Y't /TVA/c;3
5
C /c/ ra n s c
STILLMEADOW.Inc.

BLUEGREEN LAKE GUARD OXY: AN ACUTE ORAL TOXICITY STUDY WITH THE
NORTHERN BOBWHITE USING A SEQUENTIAL TESTING PROCEDURE
FINAL REPORT
EASTON STUDY NUMBER: 1004B-101
eSM PROJECT NUMBER: S23-201659
TEST GUIDELINE
OECD 223(2016)
AUTHORS
Patrick M. Hubbard,B.S.
Diana L. Temple, M.S.
STUDY INITIATION DATE: October 30,2023
STUDY COMPLETION DATE: February 23,2024
SPONSOR
BlueGreen Water Technologies
2233 NW 41'Street Suite 4200-D
Gainesville,Florida,32606
USA
TESTING FACILITY
Eurofms EAG Agroscience, LLC
8598 Commerce Drive
Easton, Maryland 21601
USA
Page 1 of 63

STUDY NO.: 1004B-101
-2
STATEMENT OF NO DATA CONFIDENTIALITY CLAIMS
No claim of confidentiality, on any basis whatsoever, is made for any information contained in this
document. I acknowledge that information not designated as within the scope of FIFRA sec. 10(d)(1)(A),
(B), or (C) and which pertains to a registered or previously registered pesticide is not entitled to
confidential treatment and may be released to the public, subject to the provisions regarding disclosure to
multinational entities under FIFRA sec. 10(g).
Company: BlueGreen Water Technologies
Company Agent: Jessica Frost
Title: Scientific Director
Signature puace-CLg_s Date 02/23/2024
THESE DATA MAY BE CONSIDERED CONFIDENTIAL IN COUNTRIES OUTSIDE THE UNITED
STATES.

STUDY NO.: 1004B-101
-3 -
GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT
SPONSOR: BlueGreen Water Technologies
TITLE: Bluegreen Lake Guard Oxy: An Acute Oral Toxicity Study with the Northern Bobwhite
Using a Sequential Testing Procedure
STUDY NUMBER: 1004B-101
STUDY COMPLETION: February 23,2024
This study was conducted in compliance with Good Laboratory Practice Standards as published
by the U.S. Environmental Protection Agency (40 CFR Part 160)(1989), which are compatible with the
Organisation for Economic Cooperation and Development (OECD) Principles of Good Laboratory
Practice(ENV/MC/CHEM(98) 17)with the following exceptions:
The characterization and stabilityof the test substance,under conditions of storage at the test site
g
were not determined in compliance with Good Laboratory Practice Standards.
A preliminary range-finding test was considered exploratory work and was not conducted
according to Good Laboratory Practices.
Annual analyses of feed and water for potential contaminants were not conducted according to
Good Laboratory Practice Standards, but were performed using a certified laboratory and standard U.S.
Environmental Protection Agency analytical methods.
STUDY DIRECTOR:
41 -d( Fib z 3 2 o 2-y
Patrick M. Hubbard,B.S. DATE
Senior Scientist
Eurofins EAG Agroscience,LLC
SPONSOR'S REPRESENTATIVE:
�L` 02/23/2024
�t DATE
SUBMITTER:
Intentially left blank Intentially left blank
DATE

STUDY NO.: 1004B-101
-4-
QUALITY ASSURANCE STATEMENT
This study was examined for compliance with Good Laboratory Practice Standards as published
by the U.S. Environmental Protection Agency (40 CFR Part 160)(1989), which are compatible with the
Organisation for Economic Cooperation and Development (OECD) Principles of Good Laboratory
Practice (ENV/MC/CHEM (98) 17). The dates of all audits and inspections and the dates any findings
were reported to the Study Director and Laboratory Management were as follows:
DATE REPORTED TO:
ACTIVITY DATE CONDUCTED STUDY DIRECTOR MANAGEMENT
Protocol October 31,2023 October 31,2023 November 1,2023
Capsule Preparation October 31,2023 October 31,2023 October 31,2023
Observations November 14,2023 November 14,2023 November 15,2023
Data&Draft Report February 1-5,2024 February 5,2024 February 5,2024
Final Report February 22,2024 February 22,2024 February 23,2024
All inspections are study based unless otherwise noted.
(t'
/-2.3 ma y
Darryl Anderso ,B.S. DATE
Quality Assurance Associate II
Eurofins EAG Agroscience,LLC

STUDY NO.: 1004B-101
- 5-
REPORT APPROVAL
SPONSOR: BlueGreen Water Technologies
TITLE: Bluegreen Lake Guard Oxy: An Acute Oral Toxicity Study With the Northern Bobwhite
Using a Sequential Testing Procedure
STUDY NUMBER: 1004B-101
STUDY DIRECTOR:
�-�� Fe6 2.3 20241
Patrick M.Hubbard, B.S. DATE
Senior Scientist
Eurofins EAG Agroscience,LLC
MANAGEMENT:
Diana L.Temple,M.S. DATE
Director of Avian Toxicology
Eurofins EAG Agroscience, LLC

STUDY NO.: 1004E-101
-6-
TABLE OF CONTENTS
Title Page
Statement of No Data Confidentiality Claims 2
Good Laboratory Practice Compliance Statement 3
Quality Assurance Statement 4
Report Approval 5
Table of Contents 6
Summary 8
Introduction 9
Objective 9
Experimental Design 9
Materials and Methods 10
Test Substance 10
Test Organisms 11
Identification 11
Avian Feed and Water 11
Dose Preparation and Dosing 12
Duration of the Test 12
Housing and Environmental Conditions 12
Observations 13
Animal Body Weights/Feed Consumption 13
Necropsy 13
Statistical Calculations 13
Results 14
Mortalities and Clinical Observations 14
Body Weight and Feed Consumption 14
Necropsy 15
Summary of Results 15
References 16

STUDY NO.: 1004E-101
- 7-
TABLE OF CONTENTS
(Continued)
TABLES, FIGURES,AND APPENDICES
Table 1. Cumulative Mortality from a Northern Bobwhite Acute
Oral Toxicity Study with Bluegreen Lake Guard Oxy 17
Table 2. Summary of Clinical Signs from a Northern Bobwhite Acute
Oral Toxicity Study with Bluegreen Lake Guard Oxy 19
Table 3. Gross Necropsy Findings from a Northern Bobwhite Acute
Oral Toxicity Study with Bluegreen Lake Guard Oxy 20
Appendix 1. Study Protocol 21
Appendix I1. Diet Formulation 44
Appendix III. Dosing Regime from a Northern Bobwhite Acute Oral
Toxicity Study with Bluegreen Lake Guard Oxy 45
Appendix IV. Daily Observations from a Northern Bobwhite Acute Oral
Toxicity Study with Bluegreen Lake Guard Oxy 47
Appendix V. Individual Body Weights(g)from a Northern Bobwhite Acute Oral
Toxicity Study with Bluegreen Lake Guard Oxy 59
Appendix VI. Individual Feed Consumption(g/bird/day)from a Northern Bobwhite Acute Oral
Toxicity Study with Bluegreen Lake Guard Oxy 61
Appendix V I I. Personnel Involved in the Study 63

STUDY NO.: 1004B-101
- 8-
SUMMARY
SPONSOR: BlueGreen Water Technologies
STUDY: Bluegreen Lake Guard Oxy: An Acute Oral Toxicity Study with the Northern
Bobwhite Using a Sequential Testing Procedure
STUDY NUMBER: 1004B-101
eSM PROJECT NUMBER: S23-201659
TEST SUBSTANCE: Bluegreen Lake Guard Oxy
TEST DATES: Hatch—November 12,2022
Acclimation Start—July 9,2023
Experimental Start(OECD)—October 30,2023
Experimental Start(EPA)—October 31,2023
Experimental Termination—December 11,2023
TEST ANIMALS: Northern bobwhite(Colinas rirginianus)
AGE TEST ANIMALS: 50-54 weeks of age at the time of dosing
SOURCE TEST ANIMALS: Woodlawn Acres Hatchery&Gamebirds
925 190th Ave
Fairmont, MN 56031
NOMINAL TEST CONCENTRATIONS:
Control and Limit: 0 and 2000 mg a.i./kg bw(body weight)
Stage 2: 392,497,630,800, 1014, 1286, 1632,2070,2625,&3330 mg a.i./kg bw
Stage 3: 1281, 1626,2065, 2622,and 3330 mg a.i./kg bw
RESULTS:
The acute oral LD50 value for northern bobwhite exposed to Bluegreen Lake Guard Oxy as a single oral
dose was calculated to be 2472 mg a.i./kg body weight, with 95% confidence that it is greater than
1921 mg a.i./kg body weight. The slope of the dose response curve was 6.39 and the Chi-Square value
was 4.9. There was no-mortality at or below the 1632 mg a.i./kg body weight level.

STUDY NO.: 1004B-101
-9-
INTRODUCTION
This study was conducted by Eurofins for BlueGreen Water Technologies at the Eurofins avian
toxicology facility in Easton, Maryland. The test was conducted from October 31, 2023 to
December 11,2023. Raw data for all work performed at Eurofins and the fmal report will be filed by
study number in archives located on the Easton site.
OBJECTIVE
The objective of this study was to evaluate the acute toxicity of Bluegreen Lake Guard Oxy
administered orally to the northern bobwhite (Colinas virginianus). The protocol provided a sequential
testing procedure to optimize the placement of doses and to match the precision of the endpoint with the
precision required for hazard assessment. An LD5o value was to be calculated or it was to be demonstrated
that the LD50 value is above a limit dose, if appropriate and possible. The method had been designed to
minimize the numbers of birds used(1).
EXPERIMENTAL DESIGN
The test species, northern bobwhite, was chosen after conducting a non-GLP range-finder with
mallard at a 2000 mg a.i./kg dose level which resulted in regurgitation. This test was conducted
according to the approved protocol which was based on the sequential design OECD test guideline 223,
2016 (2). Based on available toxicity information, the test started with a limit test with a dose level of
2000 mg a.i./kg. The limit test resulted in 60%mortality. The results of the previous stages were used to
determine the levels for the subsequent stages(Table 1). Based upon the results of the limit test, Stage 2
was conducted with dose levels of 392,497, 630, 800, 1014, 1286, 1632,2070,2625,and 3330 mg a.i./kg
with one bird per level. Dose levels for Stage 3 were 1281, 1626, 2065, 2622, and 3330 mg a.i./kg with
two birds per level. Mixed sex northern bobwhite were randomly assigned to the control groups and
treatment pens using a random number sheet generated using Microsoft Excel.
Treatment Groups
Nominal Dose Pens per
Group (mg a.i./kg) Group Birds per Pen
1 0 5 1
2 2000 5 1
3 392,497,630,800, 1014, 1286, 1632, 1 1
2070, 2625,&3330
4 1281, 1626,2065,2622,&3330 2 1

STUDY NO.: 1004B-101
- 10-
Birds were acclimated to the study facility for 16-20 weeks and to the caging for at least
four weeks prior to the start of each stage. The birds were fasted for 16.3-16.6 hours prior to dosing. At
experimental start, a single dose of the test substance was orally administered by capsule into the crop of
each bird. Each bird was individually weighed and dosed on the basis of milligrams of active ingredient
of test substance per kilogram of body weight(mg a.i./kg). The control birds received an empty capsule
of the same size the birds in the treatment groups received.
From experimental start until termination, all birds were observed at least twice daily. A record
was maintained of all mortality, signs of toxicity, and abnormal behavior. Body weights were measured
individually on the day prior to dosing (Day -1) and on Days 3, 7, and 14 of each stage. Feed
consumption was determined by pen for approximately 24-hour intervals from Day 0 to 1, Day 1 to 2,and
Day 2 to 3. Average daily feed consumption was then determined from Days 3 to 7 and from Days 7 to
14.
MATERIALS AND METHODS
The study was conducted according to the procedures outlined in the protocol, "Bluegreen Lake
Guard Oxy: An Acute Oral Toxicity Study with the Northern Bobwhite Using a Sequential Testing
Procedure" (Appendix 1). The protocol was based on the sequential design OECD test guideline
223,2016 (2). The sequential approach improves the estimate of the toxicological response using fewer
birds. Birds were individually caged for improved welfare and estimation of the toxicological response.
In order to control bias, mixed sex adult birds were randomly assigned to pens using a random number
sheet generated using Microsoft Excel. No other potential sources of bias were expected to affect the
results of the study.
The test substance was administered orally by capsule. This route of administration was selected
because a precise amount of test substance can be administered for a specified body weight. This route of
exposure also correlates to acute exposure used in mammalian toxicology.
Test Substance
The test substance was received from BlueGreen Water Technologies on September 26, 2023 and
was assigned testing facility identification number 18601 upon receipt. The test substance was a solid
and was identified as: Lake Guard Oxy; Batch Number: TMP01851. The test substance had a reported

1
1

STUDY NO.: 1004B-101
- 11 -
puirty of 83.3% Sodium percarbonate and had an expiration date of April 2026. The test substance was
held under ambient and dark conditions in locked storage at the Eurofins facilities in Easton,Maryland.
Test Organisms
The northern bobwhite represents an ecologically significant and widely distributed species in the
United States and in certain areas of Europe. The bobwhite has demonstrated sensitivity to the effects
produced by known toxic chemicals and has proven to be a good laboratory species from which a large
amount of baseline data has been gathered.
All northern bobwhite(Colinas virginianus)were 50-54 weeks of age and appeared to be in good
health at experimental start. Birds of both sexes were randomly assigned,without regard to sex,to the test
pens. There were no mortalities among the birds potentially used for the study during at least the last
14 days of acclimation. The northern bobwhite ranged in weight from 198 to 243 grams at the time of
dosing. The birds were obtained from, and identified as northern bobwhite by, Woodlawn Acres
Hatchery& Gamebirds, Fairmont, MN 56031. During acclimation to the test facility and during the test,
test birds were maintained under conditions that would not facilitate reproduction. All birds were from the
same hatch,pen-reared and phenotypically indistinguishable from wild birds.
Identification
Each pen was identified with a unique alphanumeric designation,project number,and dose group.
Individual birds within each pen were uniquely identified by numbered and colored leg bands.
Avian Feed and Water
Throughout acclimation and testing, all test birds were fed a game bird ration formulated to
Eurofins specifications by Delmarva Feed and Farm Service, Kennedyville, MD (Appendix III). Feed,
was provided ad libitum during acclimation and during the test, except during the period of fasting prior
to testing and approximately one hour following dosing. Water, from the town of Easton public water
supply, was provided ad libitum. Beginning two days after arrival at the test facility, test birds were
given water soluble antibiotics in their drinking water for eight consecutive days. The birds received no
form of antibiotic medication during at least the last 14 days of acclimation or during the test. The
municipal water and basal ration used for avian studies are screened at least annually for pesticides and
metals. Specifications of acceptable levels of contaminants have not been established. However, no

STUDY NO.: 1004B-101
- 12-
analytes have been measured at levels that were expected to have an impact on the study. Reports for the
latest analyses are stored in the archives at the Eurofms EAG Agroscience,LLC Easton site.
Dose Preparation and Dosing
The test substance was dosed neat by gelatin capsule. The test dose and the LD50 value are
reported as milligrams of active ingredient of test substance per kilogram of body weight. The nominal
doses used in this study were 0, 392, 497, 630, 800, 1281, 1286, 1626, 1632, 2000, 2065, 2070, 2622,
2625, and 3330 milligrams of active ingredient of test substance per kilogram of body weight (mg
a.i./kg).
The birds were fasted for 16.3-16.6 hours prior to dosing. At experimental start, an empty
capsule coated with corn oil was orally intubated directly into the crop of each control bird. Birds at the
treatment levels were dosed in the same manner but with a capsule containing the appropriate amount of
the test substance. Each bird was individually weighed and dosed on the basis of milligrams of active
ingredient of test substance per kilogram of body weight(Appendix III).
Duration of the Test
The primary phases of this test and their durations were:
1. Acclimation to Test-Caging—4-6 weeks
2. Fasting—16.3-16.6 hours
3. Dosing Control and Limit—Day of Experimental Start—October 31,2023
4. Dosing Stage 2—November 14, 2023
5. Dosing Stage 3 —November 27,2023
6. Post-dosing Observation— 14 days
Housing and Environmental Conditions
During the test, birds were individually housed indoors in batteries of pens(GQF Manufacturing
Co.,GQF Model No. 0315). Each pen measured approximately 25 X 51 cm. The pens had sloping floors
that resulted in ceiling height ranging from 20 to 26 cm. External walls, ceilings, and floors of each pen
were constructed of coated wire mesh. Side walls were galvanized sheeting. Each test pen contained one
bird. Birds were randomly assigned to pens.
Birds were maintained at ambient room temperature and humidity. The continuously monitored
average temperature for this study was 19.8°C with a range 17.1-21.6°C. The continuously monitored
average relative humidity was 42% with a range of 16-70%. The air handling system in the study room

STUDY NO.: 1004B-101
- 13 -
was designed to vent up to 15 room air volumes every hour and replace them with fresh air. The air
handling system was on and functional during the test. The photoperiod(maintained by a time clock)was
approximately eight hours of light per day/16 hours dark during acclimation and throughout the test. The
light source was fluorescent lights that closely approximated the color spectrum of noonday sunlight. The
birds were exposed to an average of approximately 212 lux of illumination. Housing and husbandry
practices were conducted so as to adhere to guidelines established by the National Research Council(3).
Observations
During at least the last 14 days acclimation, all birds were observed at least daily. Birds
exhibiting abnormal behavior or physical injury were not used. Following dosing, multiple observations
were performed on Day 0 of the test, including a period of at least 120 minutes post-dosing with
particular attention being paid for signs of regurgitation. All birds were observed at least twice daily
during the remaining days of the test. A record was maintained of any mortality, signs of toxicity, and
abnormal behavior.
Animal Body Weights/Feed Consumption
Body weights were measured individually on the day prior to dosing (Day -1) and on Days 3, 7,
and 14 of each stage. Feed consumption was determined by pen for approximately 24-hour intervals from
Day 0 to 1, Day 1 to 2, and Day 2 to 3. Average daily feed consumption was then determined from Days
3 to 7 and from Days 7 to 14. Feed consumption was determined by measuring the change in weight of
the feed presented to the birds over a given period of time. The accuracy of feed consumption values may
have been affected by the unavoidable wastage of feed by the birds. Measured and calculated values
presented serve as estimates of feed consumption.
Necropsy
A gross necropsy was performed on each mortality. A gross necropsy was also performed on the
carcasses of each bird from the control group and from each surviving treatment bird at the end of each
stage of the test. A gross necropsy included,but was not limited to, a general examination of the exterior
of the bird and an examination of the thoracic and abdominal cavities, including cardiovascular and
respiratory systems,liver, spleen,gastro-intestinal tract,and urogenital system.
Statistical Calculations
Mortality data were used to calculate an LDso value using the computer program SEDEC (4).
The LD5o value and 95%confidence limits were calculated using the probit method.

STUDY NO.: 1004B-101
- 14-
RESULTS
Mortalities and Clinical Observations
No regurgitation was noted among the control birds or among any of the treatment birds. There
were no mortalities in the control group or at the 392,497,630,800, 1014, 1281, 1286, 1626, 1632,2065,
2070,and 2625 mg a.i./kg dose levels(Table I). There was 60%(3 of 5)mortality at the 2000 mg a.i./kg
level, 67% (2 of 3) at the 3330 mg a.i./kg level, and 100% (2 of 2) at the 2622 mg a.i./kg level. All
control birds were normal in appearance and behavior throughout the test. All birds from the 392, 497,
800, 1014, 1281, 1286 and 2070 mg a.i./kg levels were normal in appearance and behavior throughout the
test.
Five of the twenty-five treatment birds were noted with white froth around their mouth during
observations following dosing on Day 0 of the test. Signs of toxicity were noted at the 630, 1626, 1632,
2000, 2622, 2625, and 3330 mg a.i./kg levels. The signs of toxicity noted were ruffled appearance,
lethargy, lower limb weakness, loss of coordination, reduced reaction, depression and prostrate posture.
The mortalities that occurred were noted on Days 0, 1, 7, 9, and 12 of the test. A summary of clinical
signs and mortality is presented as Table 2. Daily observations are presented in Appendix IV.
Body Weight and Feed Consumption
From Day-1 to Day 3, all remaining birds in the treatment groups had a reduction in body weight
that was greater than the control group reduction. From Day 3 to Day 7, the birds in the 630, 1014, and
2070 mg a.i./kg levels and some of the remaining birds in the 1626, 2000, and 2065 mg a.i./kg levels
continued to have a weight loss that was greater than any control bird for that interval. From Day 7 to
Day 14, all remaining birds in the treatment groups, except one each in the 1626 and 2000 mg a.i./kg
levels,had gains in body weight that were comparable to or greater than the control group(Appendix VI).
From Day 0 to Day 3, at at least one interval, the remaining birds in the treatment groups had a
feed consumption measurement that was reduced when compared to the control group. From Day 3 to
Day 7, feed consumption continued to be reduced and less than any control bird (<11g)for some of the
remaining treatment birds. From Day 7 to Day 14, the only remaining treatment birds that still had a
reduced feed consumption measurement were one bird each at the 1626 and 2065 mg a.i./kg treatment
levels(Appendix VI).

STUDY NO.: 1004B-101
- 15-
Necropsy
A gross necropsy was performed on each mortality. All mortalities were noted with remarkable
findings. Some common findings were, loss of muscle mass, emaciated, prominent keel, whitish froth
around mouth, pale liver, pale spleen, pale kidneys, gelatinous film around crop, petechial hemorrhaging
in crop tissue,'scar tissue around crop,and a primarily empty gastro-intestinal tract(Table 3).
A necropsy was performed on all birds from the control group and all surviving birds from
gross
the treatment levels. There were no findings for any of the control birds necropsied at test termination. Of
the surviving treatment birds, two were noted with remarkable findings. The bird from pen A99 in the
1626 mg a.i./kg level was noted with a loss of muscle mass and the bird from pen A102 in the 2065 mg
a.i./kg treatment level was noted with a pale,mottled, and enlarged liver.
SUMMARY OF RESULTS
The acute oral LD50 value for northern bobwhite exposed to Bluegreen Lake Guard Oxy as
a single oral dose was calculated to be 2472 mg a.i./kg body weight,with 95%confidence that it is greater
than 1921 mg a.i./kg body weight. The slope of the dose response curve was 6.39 and the Chi-Square
value was 4.9. There was no-mortality at or below the 1632 mg a.i./kg body weight level.

STUDY NO.: 1004B-101
- 16-
REFERENCES
1. Organization for Economic Cooperation and Development. 1996. Final Report of the OECD
Workshop on Harmonization of 6"alidalion and Acceptance Criteria for Alternative Toxicological Test
Methods. 22-24 January, 1996. Solna,Sweden.
2. Organization for Economic Cooperation and Development. 2016. Avian Acute Oral Toxicity Test.
OECD Guideline for Testing of Chemicals. Guideline 223. July,2016.
3. National Research Council. 2011. Guide for the Care and Use of Laboratory Animals. 8h edition.
Washington,D.C. National Academy Press. 220 pp.
4. Springer,T.A., 10/5/2009. Sequential Design Calculator:A Tool for Use with OECD TG 223 Avian
Acute Oral Toxicity Test, Users Guide. SEDEC Version 1.3. OECD,Paris.

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STUDY NO.: 1004B-101
- 19-
Table 2
Summary of Clinical Signs from a Northern Bobwhite Acute Oral
Toxicity Study with Bluegreen Lake Guard Oxy
Nominal Dosage Clinical Signs Last Day with Day of Percent
Stage (mg a.i./kg) First Noted Clinical Signs Clinical Signs Mortality Mortality
Limit 2000 56 min 1,2,4,11,13,14 12 0,9,& 12 60
Stage 2 392 NA NA NA NA 0
497 NA NA NA NA 0
630 Day 7 11 Day 9 NA 0
800 NA NA NA NA 0
1014 NA NA NA NA 0
1286 NA NA NA NA 0
1632 Day 1 11 Day 2 NA 0
2070 NA NA NA NA 0
2625 Day 1 11 Day 2 NA 0
3330 3hr 24 min 4,11,13 Day 0 Day 0 100
Stage 3 1281 NA NA NA NA 0
1626 2 hr 17 min 11,14 14 NA 0
2065 NA NA NA NA 0
2622 3hr32min 4,5,11,14 6 1 &7 100
3330 3 hr 32 min 1,5,11,14 3 0 50
NA=Not applicable
*Clinical signs:
1 =depression.2=reduced reaction,4=loss of coordination,5=prostrate posture.11 =ruffled appearance.
13=lower limb weakness, 14=lethargy

STUDY NO.: 1004B-101
-20-
Table 3
Gross Necropsy Findings from a Northern Bobwhite Acute Oral
Toxicity Study with Bluegreen Lake Guard Oxy
Treatment groups(mg a.i./kg) 1626 2000 2000 2000 2065 2622 2622 3330 3330
Pen A99 A37 A40 A41 A102 A103 A104 A76 A106
Mortality no yes yes yes no yes yes yes yes
Loss of muscle mass X X X - - X - - -
Emaciated - X X - - - - - -
Prominent keel - X X - - X - - -
Whitish froth around mouth - - - X - - - X -
Liver pale - X X - X - - - -
Liver mottled - - - - X - - - -
Liver enlarged - - - - X - - - -
Spleen pale - X X - - X - - -
Kidneys pale - - X - - X - - •
Gelatinous film around crop - - - - - - X X X
Petechial hemorrhaging in crop tissue - - - - - - X X X
Scar tissue around crop - - X - - X - - -
Crop tissue scared with areas of - X - - - - - - -
necrosis
GI tract primarily empty - X X - - X - - -
Birds not listed above had no remarkable findings.

STUDY NO.: 1004B-101
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Appendix 1
Study Protocol

STUDY NO.: I004B-IOl
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PROTOCOL
BLLBGREEN LAKE GUARD OXY: AN ACUTE ORAL TOXICITY STUDY WITH
THE NORTHERN BOBWHITE USING A SEQUENTIAL TESTING PROCEDURE
OECD Guideline 223
Submitted to
BlueGreen Water Technologies
2233 NW 41'Street Suite#200-D,
Gainesville,Florida,32606 USA
Testing Facility
Eurotins EAG Agroscience,LLC
8598 Commerce Drive
Easton,Maryland 21601
1410)822-8600
October 24,2023

STUDY NO.: 1004B-101
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BLUEGREEN LAKE GUARD OXY: AN ACUTE ORAL TOXICITY STUDY WITH
THE NORTHERN BOBWHITE USING A SEQUENTIAL TESTING PROCEDURE
SPONSOR: BlueGreen Water Technologies
2233 NW 41"Street Suite k200-D,
Gainesville, Florida, 32606 USA
SPONSOR'S REPRESENTATIVE: Jessica Frost
[e-mail]
TESTING FACILITY: Eurofins EAG Agroscience, LLC
8598 Commerce Drive
Easton, Maryland 21601 USA
STUDY DIRECTOR: Patrick M. Hubbard, B.S.,Senior Scientist
Eurofins EAG Agroscience, LLC
LABORATORY MANAGEMENT: Diana Temple, M.S.
Director of Avian Toxicology
FOR LABORATORY USE ONLY
Proposed Dates:
Experimental Experimental
Start Date: Cc 4. 5/ i o z' Termination Date: No. f y,
Easton Study No.: 1004B-101 (eSM Project No.: S23-201659)
Test Concentrations: O .,,f Z.o oo
Test Substance No.: 18 Co/ Reference Substance No.:(if applicable): —'-
PROTOCOL APPROVAL.
STUDY DIRECTOR DATE
LABORATORY MANAGE NT DATE
SP SOR'S REP • DA '
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•
STUDY NO.: 1004B-101
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OBJECTIVE
The objective of this study is to evaluate the acute toxicity of a test substance administered orally
to the northern bobwhite(Colinas virginianus). This protocol provides a sequential testing procedure that
optimizes the placement of doses and matches the precision of the endpoint with the precision required
for hazard assessment. An LDso value will be calculated or it will be demonstrated that the LD,,,value is
above a limit dose,if appropriate and possible. The method has been designed in a way to minimize the
numbers of birds used.
SUMMARY
Dosages of the test substance will be administered to groups of northern bobwhite based on the
known or suspected toxicological properties of the test substance. Since the information required by
different hazard assessment schemes may vary considerably,two alternative tests may be performed:
• Limit-Dose test—this is the preferred test when toxicity is expected to be low and lethality is
unlikely at the limit dose. The limit dose must be adequate for assessment purposes,and it is
usually at least 2000 mg/kg body-weight or the maximum expected residue level(whichever
is higher). In the Limit-Dose test,five birds will be dosed at the limit dosage. The test may
be extended to a Dose-Response test depending on the outcome of the Limit-Dose test
(Figure 1).
• Dose-Response test—this is the preferred test when the slope of the dose response curve
andior the confidence interval is required in addition to an estimate of the LDso.
Doses for the first stage of the Dose-Response test may he based on range finding,the outcome of
a Limit-Dose test (Table I) or other available toxicity data(e.g. rat LDso value). A progression of
dosages,based upon response to the previous dosage,will be used for the Dose-Response test(Figure 2).
Dosed birds will be observed for toxicological responses for at least three days(or up to 14 days.based
upon response)in order to determine the next dosage. Dosages will be adjusted based upon the response
of each group of birds dosed. Depending on the needs for risk assessments the sponsor may decide not to
proceed with further steps. At least five untreated control birds from the same lot will be included in the
test. Control birds will be sham dosed with the same capsule(s)or carrier used with the test substance,
and will be maintained under the same conditions as treated birds. Sham dosing will be performed on the
same day as the first dosing with test substance(either with the Limit-Dose test,if it is performed.or with
Stage 1 of the Dose-Response test). Additional control birds may be added at a later date,if needed.
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MATERIALS AND METHODS
The methods,species used and route of administration are based upon procedures specified in the
OECD guideline 223.Avian Acute Oral Toxicity Test(1). For this study the northern bobwhite was
chosen as the test species after a non-GLP preliminary rangefinder was conducted with mallards and
regurgitation was noted. For a given northern bobwhite study, the route of administration may be
determined duringrange findingtests conducted accordingto Standard Operating Procedure(SOP)417.
g P g
In order to control bias,adult birds will be randomly assigned to pens. Any impact of testing multiple
dosage groups over a period of time will be mitigated by the use of birds from the same population and
the consistency of husbandry practices. No other potential sources of bias are expected to affect the
results of the study.
The test substance will be administered orally, by capsule or gavage. These routes of
administration were selected because a precise amount of test substance can be administered for a
specified body weight. In the case of granular formulations,this method best represents actual exposure
under field conditions. This route of exposure also correlates to acute exposure used in mammalian
toxicology.
Test Substance
Information on the characterization of test,control or reference substances is required by Good
Laboratory Practice(GLP)Standards and Principles. The Sponsor is responsible for providing the testing
facility with verification that the test substance has been characterized according to GLPs prior to its use
in the study. If verification of the GLP test substance characterization is not provided to the testing
facility.it will he noted in the compliance statement of the final report.
The Sponsor is responsible for all information related to the test substance and agrees to accept,
or give the testing facility authorization to dispose of any unused test substance and/or test substance
containers remaining at the end of the study.
Test Birds
The northern bobwhite(Celina% virginianier) represents an ecologically significant and widely
distributed species in the United States and in certain areas of Europe. The bobwhite has demonstrated
sensitivity to the effects produced by known toxic chemicals and has proven to be a good laboratory
species from which a large amount of baseline data has been gathered.
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The bobwhite will be obtained from and identified by Trace Pheasantry,Douglassville,PA 19518
or another reputable supplier. Each lot of birds is assessed by trained personnel upon receipt. If
mortalities that appear to he non-incidental occur during acclimation in birds received from a supplier,a
necropsy of representative individuals may be performed in order to determine the cause of death.
Deformed abnormal,sick or injured birds will not be used. Birds will not be used for a test if more than
5 percent of the total test population die during the 14-day acclimation period preceding the test.
All bobwhite will be from the same hatch,at least sixteen weeks of age and apparently healthy at
initiation of the test. Depending upon age and strain,birds will range in weight from 160-250 grans. All
birds within a given test will he indiscriminately selected with regard to sex. All test birds will be
acclimated to the test cages for at least 14 days prior to the initiation of the test and at least the 14 days of
acclimation records will be maintained with the study data. The birds used in the test will be uniquely
identified by numbered leg bands. Groups of test pens will be identified by study and dosage.
Treatment Regimes
Limit-Dose Test and Possible Extension to Dose-Response Test
Dosing will begin with the Limit-Dose Test,unless available information suggests that
the LD5„is below the limit dose(typically considered to be 2000 rng/'kg,unless a higher dose is
necessary to meet risk criteria). The Limit-Dose test design consists of dosing five animals at the
limit dose. Birds are given a single oral dose of the test substance(mg/kg body weight)into the
crop,and then observed for 14 days.
I. If no mortality occurs,it can be concluded at the 95%confidence level,that the LD,,,
is above the limit dose.
2. If only five birds were initially dosed and one treatment related death is observed,if
no signs of toxicity are observed in other hinds,then five more birds may be dosed at
the limit,or at the discretion of the study director,the test may proceed to Stage 2 of
the Dose-Response test design. If there is only one death in the total of 10 birds,then
it can be concluded that at the 95%confidence level,the LD,c is above the limit dose
and no further testing is required. If more than one bird in the total of 10 dies Urea
Stage 2 of the Dose-Response test is executed(Figure 2 and Table 3).
3. If the observed mortality is I out of 5 birds and there arc signs of toxicity in other
birds,or if there are between 2 and 4 mortalities among 5 birds,Stage 2 of the Dose-
Response test will be executed.
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5test(starting
If mortalityis complete(i.e.all birds have died),the
P Dose-Response
with Stage l) design will be used. If needed,additional dosing stages can begin
before the 14-day period of observation is complete.
To proceed from the Limit-Dose test to Stage 2 of the Dose-Response test method,an
initial estimate of the LD50 is needed to select doses at this stage. Working estimates for this use
arc given in Table 1. Using the working estimate of the LDan,the lowest and highest doses can be
calculated as described in the directions for Stage 2. The calculated high dose (hdose) will
generally need to be adjusted(reduced)to the highest level that is consistent with practicality and
animal welfare constraints. It should be noted that in some circumstances,the LDS,can not be
estimated without using doses above the limit dose. Because there are constraints on the use of
very high doses of test substance,it may not always he possible to estimate the LDS)for slightly
toxic substances.
Dose-Response Test
The Dose-Response test employs staged test designs where animals are dosed at different
times(stages)to allow the use of accumulated information to optimally position doses along the
dose response curve.
At each stage.one or more birds are given a single oral dose(mg,kg body weight)of the
test substance into the crop. All birds are observed for 14 days, but selection of doses for
subsequent stages is typically based on results after 3 days. This interval may be reduced if birds
quickly show signs of recovery or extended if delayed mortality is expected or observed. If
delayed mortality occurs in early stages after dose selection for second or third stages.placement
of doses at later stages will not be optimal. If the Limit-Dose test is not used as the 1st dosing
stage,each of at least four birds is given a different dose,selected to bracket the best available
estimate of the LD50 (e.g. based on the rat or other bird species' LDso or QSAR estimates).
Depending on the outcome of Stage 1,the doses for Stage 2 arc determined(see Table 2). The
process continues to Stage 1 and possibly Stage 4,in the Dose-Response test. Observations of
deaths that are clearly not treatment related(c.g.due to physical injury)should be excluded from
calculations.
Control Group
At least five untreated control birds from the same lot will be included in the test.
Control birds will be sham dosed with deionized water if the test substance is dosed neat or with
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the same capsules)or carrier used with the test substance,and will be maintained under the same
conditions as treated birds. Sham dosing will be performed on the same day as the first dosing
with test substance (either with the Limit-Dose test, if it is performed, or with Stage 1 of the
Dose-Response test). Additional control birds may be added at a later date, if needed(e.g. if
dosing volume or number of capsules changes to accommodate higher dosages). When
appropriate and possible and with Sponsor approval,a common control group may be used. If a
common control group is used,the original control data will be permanently maintained in the
archives at the Easton site as a facility record.
Duration of the Test
The primary phases of this test and their durations are:
1. Acclimation-At least 14 days.
2. Fasting—At least 15 hours.
3. Dosing—For each stage.a single dose on Day 0 of the stage.
4. Multiple Stages Except in the case of total mortality,a minimum of three days between
stages.
5. Post-dosing observation-At least 14 days for each dosing stage.
Animal Diet
All test birds will be fed a game bird ration formulated to the testing facility's specifications
(Appendix 1). Feed and water will be provided ad libitum during acclimation and during the test,except
during periods of fasting. The water source will be the Town of Easton public water supply. The birds
will be fasted for a minimum of Ii hours prior to dosing. The birds will receive no form of antibiotic
medication in the diet during the test.
Feed and water arc analyzed at least annually for selected metals. organics and pesticides in
accordance with the testing facility Standard Operating Procedures. Specifications for acceptable levels
of contaminants in rations for avian species have not been established. However,there are no levels of
contaminants reasonably expected to be present in the diet that are considered to interfere with the
purpose or conduct of the study.
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Dosage Preparation and Dosing
Choice of the method for presentation of the test substance will be determined prior to the start of
the study and. may be based upon pre-test method trials. The method chosen will be consistent for all
dosage groups and the control group. The test substance may be administered using a gelatin capsule or
capsules.Prior to dosing,each bird will be weighed. The appropriate amount of test substance for each
individual bird will be weighed and placed in a gelatin capsule(s). The capsule(s)will be administered to
the bird at the approximate level of the crop. A record of the amount of test substance in each capsule
given to each bird will be maintained in the raw data.
Alternatively,the test substance may be dosed neat or dissolved or suspended in a suitable diluent
(deionized water by preference.or table grade corn oil,0.5%carboxymethylcellulose or propylene glycol,
if solubility dictates). The concentration of the test substance in the diluent will be adjusted to give an
approximately constant volume to body weight ratio. At initiation of the tcst,a single dose of the test
substance in diluent will be orally intubatcd directly into the crop of each bird using a stainless steel
catmula. Each bird will be individually weighed and dosed on the basis of milligrams of test substance
per kilogram of body weight. The maximum amount of diluent used will not exceed 1%of the body
weight of the test birds.
Control birds will receive an empty gelatin capsule(s)of the same size utilized among the birds
receiving the test substance or a volume of diluent equal to the largest volume to body weight ratio
utilized among the birds receiving the test substance,as appropriate. All test substance calculations will
be based on the test substance as received or corrected to 100%active ingredient based on the information
provided by the Sponsor.
Dosing Solution Sampling and Analysis
If utilized,samples of the dosing solutions will nut be collected for chemical analysis.
Housing and Environmental Conditions
Housing and husbandry practices will be conducted so as to adhere to the principles established by
National Research Council(2). Test birds will be housed in battery breeding pens manufactured by GQF
Manufacturing Company(Model No.206 or 0330)or equivalent. Each pen has floor space that measures
approximately 25 X 51 cm. Floors arc sloping so that ceiling height will range from approximately 20 to 26
cm. External walls,ceilings and floors of each breeder pen are constructed of wire mesh while side walls
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are constructed of galvanized sheeting. Each pen will contain one bird. Pens will be labeled
alphanumerically and groups of test pens will be labeled by study numberis)and dosage level.
Birds will be maintained at an ambient temperature of 15-30°C. Ambient temperature and
relative humidity will be recorded at least twice daily. An 8 hours lighdl6 hours dark photoperiod will be
maintained. Light intensity will be measured at least once during the test in accordance with the testing
facility Standard Operating Procedures. The average will be based upon light intensity measurements in
each pen. Lighting will be provided by fluorescent lights that closely approximate noonday sunlight(noon-
day sun-4870,Chroma 50 or equivalent-5000 Kelvin').
Observations
During the test, all birds will be observed at least once daily for mortality and clinical signs.
Birds arc observed repeatedly on the day of dosing.with particular attention paid to signs of regurgitation.
and at least twice daily until test termination. If signs of toxicity or mortality persist to Day 14 of a given
stage,the observation period may be extended for that stage.
animal Body Weights
Individual body weights will be measured at initiation of the test stage and on Days 3.7,and 14
of the stage. In the case of capsule dosing,body weights may be taken on the day prior to dosing to allow
time for capsule preparation. If the stage is extended,body weight will be measured at stage termination.
If the extension is longer than seven days,body weight measurements will be performed weekly and at
stage termination. For body weight determination,birds arc caught by knowledgeable,trained personnel.
The key to minimizing stress on the bird is to capture the bird efficiently but gently.and to hold the bird
"caged"in the hand so as to not allow escape,but to minimize pressure.
Feed Consumption
Estimated feed consumption will be determined by pen for each dosage group and the control
group for approximately 24 hr intervals from Day 0 to Day I,Day I to Day 2 and Day 2 to Day 3. Feed
consumption will then be measured from Day 3 to Day 7 and from Day 7 to Day 14. If the stage is
extended,feed consumption will be measured at stage termination. If the extension is longer than seven
days, feed consumption measurements will be performed weekly and at stage termination. Feed
'General Electric Institute,General Electric Company,Ncla Park,Cleveland,Ohio.
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consumption will be determined by measuring the change in the weight of the feed presented to the birds
over a given period of time. Given the nature of the feed presented to the birds and their wasteful behavior,
the accuracy of feed consumption values may be affected by the unavoidable wastage of feed by the birds.
Necropsy
Necropsies will be performed under those circumstances where the study director feels that
necropsies may yield information relevant to the interpretation of the study. Typically,two or three birds
are examined from those dosage levels experiencing mortalities and all control mortalities are examined.
Gross necropsies will include,but are not limited to.a general examination of the exterior of the bird and
an examination of the thoracic and abdominal cavities,including cardiovascular and respiratory systems,
liver,spleen,gastro-intestinal tract,and urogenital system. Findings relevant to the interpretation of the
study will be reported.
Euthanasia of Test Birds
At test termination,test birds will be euthonized by using carbon dioxide gas or other appropriate
methods that ensure a humane death, according to recommendations in AVMA guidelines (3). The
method used will be documented in the raw data.
Statistical Calculations
An LDS„value along with 95%confidence limits will be calculated, when possible using data
accumulated from all test stages. The data will be analyzed.in order of preference,by probit analysis,
moving average or binomial probability(4.5,6,7). The choice of method used for calculating the LD,,
value will he based upon the mortality pattern observed(8,9,10).
RECORDS TO BE MAINTAINED
Records to be maintained for data generated at the testing facility will include,but not be limited
to:
1. The signed protocol.
2. Identification and characterization of the test substance,if provided by the Sponsor.
3. Dates of initiation and termination of the test.
4. Animal history.
5. Husbandry and environmental conditions.
6. Dosage calculation,preparation and administration.
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7. Body weight measurements.
8. Feed consumption measurements.
9, Daily observations.
10. Necropsy findings,if applicable.
I I. Statistical calculations.
12. The final report.
FINAL REPORT
A report of the results of the study will be prepared by the testing facility. The report will
include,but not be limited to,the following:
I. Name and address of the facility performing the study.
2. Dates on which the study was initiated and completed. It is the responsibility of the Sponsor to
provide the fmal date that data arc recorded for chemistry. pathology and/or supporting
evaluations that may be generated at other laboratories.
3. A statement of compliance signed by the Study Director addressing any exceptions to Good
Laboratory Practice Standards.
4. Objectives and procedures stated in the approved protocol.including any changes in the original
protocol.
5. Statistical methods employed for analyzing the data, when applicable. A description of the
transformations,calculations or operations performed on the data,a summary and analysis of the
data,and a statement of the conclusions drawn from the analysis. Results of the analysis of data
will include the calculated Ulm value.95 percent confidence limits, slope of the transformed
dose-response line,and the results of a goodness-of-fit test(e.g.X'test).
6. The test,control and reference substances identified by name,chemical abstracts number or code
number,strength,purity,and composition or other appropriate characteristics,if provided by the
Sponsor.
7. Stability and,when relevant to the conduct of the study,the solubility of the test,control and
reference substances under the conditions of administration.if provided by the Sponsor.
8. A description of the test system used. Where applicable,the final report shall include the number
of animals used,body weight range,source of supply,species(including scientific name),age,
and procedure used for identification.
9. A description of the dosage,dosage regimen.route of administration.and duration.
10. A description of the methods used,including but not limited to:
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a) Detailed description of the basal diet, including source,and supplements(if used). A
nutrient analysis of the diet will be included in the test report.
b) The number of dosages used, nominal dosage in each level, and number of birds per
dosage. Number of replicates per dosage,method and time of administration. Results of
range-finding test(if conducted).
c) Acclimation procedures and methods of assigning birds to test pens.
d) Frequency,duration and methods of observation. Description of signs of intoxication and
other abnormal behavior, including time of onset, duration,severity(including death),
and numbers affected at each dosage level and controls each day of the test period.
c) Description of housing conditions, including type. size and material of pen. room
temperatures,approximate test room humidity.photoperiod and lighting intensity.
0 Individual body weights (or means, extremes, and an estimate of variance) will be
reported for the beginning of the test and at least weekly thereafter.
g) Feed consumption by pen for Days 0-I, 1-2, 2-3. 3 to 7, 7 to 14 and any extension
periods.
h) Results of gross pathological examination,if performed.
11. A description of all circumstances that may have affected the quality or integrity of the data.
Anything unusual about the test, any deviation from these procedures, and any other relevant
information.
12. The name of the Study Director.the names of other scientists or professionals,and the names of
all supervisory personnel.involved in the study.
13. A description of the transformations, calculations, or operations performed on the data, a
summary and analysis of the data,and a statement of the conclusions drawn from the analysis.
14. The signed and dated reports of each of the individual scientists or other professionals involved in
the study.if applicable.
15. The location where all specimens,raw data,and the final report are to be stored.
16. A statement prepared by the Quality Assurance Unit listing the dates that study inspections and
audits were made and the dates of any findings reported to the Study Director and Management.
CHANGES TO FINAL REPORT
If it is necessary to make corrections or additions to a final report after it has been accepted such
changes shall be in the form of amendment by the Study Director. The amendment should clearly
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identify the part of the final report that is being added to or corrected and the reasons for the correction or
addition. Amendments shall be signed and dated by the Study Director.
CHANGES TO PROTOCOL
Planned changes to the protocol will be in the form of written amendments signed by the Study
Director and approved by the Sponsor's Representative. Amendments will be considered as part of the
protocol and will be attached to the final protocol. Any other changes will be in the form of written
deviations signed by the Study Director and filed with the raw data. All changes to the protocol will be
indicated in the final report.
GOOD LABORATORY PRACTICES
This study will be conducted and reported in compliance with Good Laboratory Practice
Standards as published by the U.S.Environmental Protection Agency(40 CFR Pact 1601(1989).which
are compatible with the Organisation for Economic Cooperation and Development(OECD)Principles of
Good Laboratory Practice(ENV/MC/CHEM(98) 17). A statement of compliance,signed by the Study
Director,will be included in the final report. The Sponsor will be responsible for compliance with Good
Laboratory Practices for procedures performed by other laboratories(e.g.,residue analyses or pathology).
Each study conducted by the testing facility is routinely examined by the testing facility Quality
Assurance Unit for compliance with Good Laboratory Practices,Standard Operating Procedures and the
specified protocol. Raw data for all work performed at the testing facility and the final report will be filed
by study number in archives located on the Easton site or at an alternative location to be specified in the
final report.
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REFERENCES
I. Organization for Economic Cooperation and Development. 2016. Avian Acute Oral Toxicity
Test. OECD Guidelines for the Testing of Chemicals. Guideline 223. July,2016.
2. National Research Council. 2011. Guide for the Care and Use of Laboratory Animals. gib
Edition.Washington,DC. National Academy Press. 220 pp.
3 Amercan Veterinary Medical Association. 2020. ALMA Guidelines for the Euthanasia of
Animals:2020 Edition. 121 pp.
4. Finney. D.J. 1971. Statistical Methods in Biological Assay. Second edition. Griffin Press,
London.
5. Thompson.W.R. 1947. Bacteriological Reviews. Vol II.No.2(June): 115-145.
6. Stephan. C. E. 1977. Methods for Calculating an LC50. Aquatic Toxicology and Hazard
Evaluations. Pages 65-84 in American Society for Testing and Materials,Pub.No.STP 634.
7. Stephan, C. E. 1978. U.S. EPA. Environmental Research Laboratory, Duluth, Minnesota.
Personal Communication.
8. Stephan,C.E. 1982. Personal Communication.Dated September 10,1982,Addressed to Lowell
Balmer.Chairman of ASTM Task Group on Calculating LCsos.
9. Springer,T.A., 10/512009. Sequential Design Calculator: A Tool for Use with OECD TG 223
Avian Acute Oral Toxicity Test, Users Guide. SEDEC Version 1.3. OECD,Paris.
10. The SAS System for Windows. 2002-2012. Version 9.4. SAS Institute Inc., Cary, North
Carolina.
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FIGURE I. LIMIT-DOSE TEST PROCEDURE
IDose 5 birds at limit dose
1 1
Outcome:0 death Outcome:1 death Outcome:2-4 deaths J Outcome:5 deaths
•
Sips of toxicity in Yes
survivors?
No
Choose
Dose 5 more birds
at brit
wir 7 7 V
Yes Nd Proceed to stage 2 of Proceed ro Sure I of
LDSO>Limit Mortality=I f 10 D -gap, DoetiRespoore tat
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F
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FIGURE 2. DOSE-RESPONSE PROCEDURE
Estimate 11330 based cm
prior knowledge Limit-Dose lest Ludt-Dose tact
(e.g.other studies) ar Complete mortality °r Partial morality
Stage I
4 doses;Third/dose
Control Nose=0.1414 x 1D50
(1f net mck l d in bdose-7.071 x LD50
hoot test)
5 birds stem chat I
waking LDso from Stage 1
Stage 2
LDSQoa ly 13 doses;I bird/dose
Stop after Stage 2 4 Wow-03425 x LDSO 4
Wow-231,x LD50
Waiting LD50 from Stags 1+2 Working LD50 from Stages 1 r2
when>1 reversals when<2 reversals
1
Stage 34 Stage 3b
2 doses;5 birds/dose 5 doses;;2 birds/dose
Nose-10(-t.autms.)x LD50 Idosa w 0.6205 x LDSO
hdose- O O.a1M4+1 x LDSO hdoso=1.6I 13 x LD50
v
Final 1D50 from Stages 1+2+3 Working LD50 from Stages 1+2+3b
when>1 reversal or>1 partial kill when Q reversals and Q pedal kills
Stare 4
1�, F LDS()from 5 doses;2 birdeldose
Cope,and madeac. ♦— kluse-0.6205 x LD50
tatorai Won-1.611 x LD50
PROTOCOL NO.: 1004"I02423/QLD-SEQ'I00P-409

STUDY NO.: 1004B-101
-38-
_17_
TABLE I
Working estimate of LDs0 for use in Stage 2 of the Dose-Response design
derived from mortality in a limit test at 2000 mg/kg
Mortality(°e1 10 20 30 40 50 60 70 80 90
Working
Estimate of 3606 2944 2541 2244 2000 1782 1574 1358 1 109
LD.)
PROTOCOL NO.: 1004/102423/QLD-SEQ;100P-409

p
STUDY NO.: I004B-I01
- 39-
-It;_
TABLE 2
Calculation of the working LDs,;computed from the four doses in Stage 1'1'
dosel dose2 dose3 dose4 Working LDso(Stage I)
O 0 0 0 (dose4 x dose5)'=
O 0 0 X (dose3 x dose4)'2
O 0 X 0 (dose2 x dose3 x dose4 x dose5)'4=(dose3 x dose4)'
O X 0 0 (closet x dose2 x dose4 x dose5)"=dose3
X 0 0 0 (dose°x dosel x dose4 x dose5)'4=Idose2 x dose3)'2
O 0 X X (dosc2 x dose3)'2
O X X 0 (dosel x dosc2 x dose4 x dose5)'4=dose3
X X 0 0 (dose°x dosel x dose4 x dose5)'4=(dose2 x dose3)'2
O X 0 X (dose!x dose2 x dose3 x dose4)''_(dose2 x dose3)'
X 0 X 0 (dose°x dosel x dose2 x dose3 x dose4 x dose5)'
—(dosc2 x dose3)'
i 4�
X 0 0 X (dose°x dose' x dose3 x dosc4) dose_2
O X X X (dose] x dose2)'=
X 0 X X (dose°x dosel x dose2 x dose3)'4=(dosel x dose2)'-
X X 0 X (dose°x dose!x dose3 x dose4)'4=dose2
X X X 0 (dose°x dosel x dose4 x dose5)'4=(dose2 x dose3►''
X X X X (dose°x dose])''
° Survival is represented by 0 and death by X.
'' Even though only tour doses(dose I through dose 4)are used in the test.values for dose 0 and dose 5
are used in the table. The values that should be used for these doses arc one step up or down from the
actual test doses. That is,dose 0=dose 1/step and dose 5=dose 4 x step. Dose 0 must be added to
the computation of the approximate LDso when mortality occurs at the lowest test dose,and dose 5 is
added when there is survival at the highest test dose.
PROTOCOL NO.: 1004/102423/QLD-SEQ/100P-109

STUDY NO.: 1004B-101
-40-
- IQ-
TABLE 3
The Dose-Response Testing Procedure Design
The following parameters are used in this section.
Idose the lowest dose used during a particular stage(mg/kg body weight).
hdose the highest dose used during a particular stage(mg/kg body weight).
Step the multiplication factor used in calculating the individual closes.
Stage I-Four doses equally spaced on a log scale around the initial estimate of the LD.,,,
The calculations for!dose,hdose,and step for Stage 1 are as follows:
(1) Calculate
/dose=dosel=0.1414 x(initial estimate of LD50)and
hdose=dose4=7.071 x(initial estimateof LD50)
(2)If hdose is greater than 3330 then set hdose=3330(may be set lower if limited by physical
constraints)and recalculate the lowest dose as!dose=hdose/50
(3)Calculate:step=50'
(4)Calculate the second and third doses:
dose 2=!dose x step.dose 3=dose2 x step=!dose x step x step
(5)Give one dose(dose),dose2,dose3,dose4)to each of the four birds.
(6)Observe birds for a period(typically three days), and note whether each bird is dead or alive.
Compute a working estimate of the LD50 as the geometric mean of the doses that produce a
transition from survival to death(sec Table 2).This working estimate of the LDs„will be used to
establish doses for Stage 2.
(7) Continue to observe the birds for eleven additional days(14 days in total).
Stake 2-Tett doses equally spaced on a lob scale around the working estimate of the Ulu obtained
from Stage I or a(link test with partial mortality
The calculations of/dose,hdose and step for Stage 2 arc as follows:
(1) Calculate: !dose=dose!=0.3425 x working LD50 and
hdasc=dasel0=2.919 x working LD50
(2) If hdose is greater than 3330 then set hdose-3330(may be set lower if limited by physical
constraints)and recalculate the lowest dose as!dose=hdose/8.5
(3) Calculate: step=(hdose I/dose)'
(4) Calculate the eight intermediate doses: dace; =/dose x step")•for i=2 to 9
(5) Give each of the ten birds one of the calculated doses.
PROTOCOL NO.: 1004/102423/QLD-SEQ;IOOP-i09

r
STUDY NO.. 1004B-101
-41 -
-20-
(6)If an LD,-only test is being conducted,observe the birds from a total of 14 days. Estimate the
LOso with the probit model using final data from Stage 1 and Stage 2.
(7) If an LD50-slope test is being run,observe the birds for a period(typically three days).and note
whether each bird is dead or alive.The number of reversals are counted and a probit model is
fitted to the combined mortality data from Stages I and 2 to obtain a working estimate of the LDs0
and,if possible.the slope. If two or more reversals are observed proceed to Stage 3a.otherwise
proceed to Stage 3b.
(8) Continue to observe the birds for an additional eleven days(14 days in total).
Stage 3a—Two doses equally spaced on a log scak around the working estimate of the Wa,obtained
from Stages I and 2
The calculations of/dose.hdose and step for Stage 3a arc as follows:
(I) Calculate: !dose=I0'"r O16'""'"g'"i0 'x working LD50
and hdose=10'''036"""t*"&°`'x working LD50
(2) If the estimate of slope from Stage 1 and 2 data is greater than fifteen or less than one,then set the
slope to fifteen or one respectively. If an estimate of the slope was not obtained from Stage I
and 2 data,assume it to be 5.
(3) If hdose is greater than 3330 then set hdose = 3330 (may be less if limited by physical
constraints)and recalculate the lowest dose as !dose=hdose I10'"'Z'''''
(4) Give each dose to five birds.
(5) Observe the birds for 14 days and note whether each bird is dead or alive. Fit a probit model to
the combined mortality data from Stages 1,2 and 3a,in order to obtain final estimates of both the
LI)so and the slope. The test is completed.
(6) If no probit model can be fitted,report this and estimate the LDx,as the geometric mean of the
concentrations on either side of the region where 50'%mortality occurs. Alternatively other
interpolation and moving average methods may be used to estimate the LD50.
Stage 3b—Five doses equally spaced on a log scale around the working estimate of the LDn,obtained
fmm Stages 1 and 2
The calculations of/dose.hdose and step for Stage 3b are as follows:
(I) Calculate: Idace=dose!=0.6205 x mt ot*ing LD50 and
hdose=dase5=1.6113 x working LD50
(2) If hdose is greater than 3330 then set hdose=3330(may be less if limited by physical constraints)
and recalculate the lowest dose as/dose=hdose/2.6
(3) Calculate: step=(hdose/►dose)'
(4) Calculate the three intermediate doses: dose!=Idose x step""",for i=2 to 4
(5) Give each dose to two birds.
(6) Observe the birds for a period(typically three days)and note whether each bird is dead or alive.
PROTOCOL NO.: 1004/102423/QLD-SEQ/100P-409

F
STUDY NO.: 1004B-101
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-21 -
Fit a probit model to the combined mortality data from Stages I, 2 and 3b in order to obtain a
working estimate of the LDN,and slope.
(7) If at least two reversals or at least two partial kills are observed and a working estimate of the
slope has been obtained then the study can be stopped. Continue observing the birds until the end
of the 14"day and lit a probit model to the combined data from Stages I.2 and 3b,in order to
obtain final estimates for both LD50 and slope. The test is completed.
(81 If conditions in 7 above are not satisfied proceed to Stage 4,but also continue observing the birds
until the end of the 14's day.
Stage 4—Five doses equally spaced on u log scale around the working estimate alike LDs obtained
front Stages 1,2 and 3b
The calculations of!dose.hdose and step for Stage 4 are as follows:
(I)Calculate: !dose=dose!=0.6205 x working LD50 and
hdose=dose5=1.6113 x working LD50
(2) If hdose is greater than 3330 then set hdose—3330(may be less if limited by physical constraints)
and recalculate the lowest dose as !dose=hdose 12.6
(3) Calculate: step=(hdose/ldoee)'
(4) Calculate the three intermediate doses: dosei=ldoee x step"-n,for i=2 to 4
(5) Give each dose to two birds.
(6) Observe the birds for 14 days and note whether each bird is dead or alive. Fit a probit model to
the combined data from Stages I,2,3b and 4 in order to obtain final estimates of both the LDsu
and the slope. The test is completed.
If at the end of Stage 4 no maximum likelihood estimate can be obtained report this and estimate the
LD50 as the geometric mean of the concentrations on either side of the region where 50%mortality
occurs.Alternatively other interpolation and moving average methods may be used to estimate the LDco.
PROTOCOL NO.: 1004/1024231QLD-SEQ/I00P-409

STUDY NO.: 1004B-101
-43 -
_22_
APPENDIX I. DIET FORMULATION
EUROFINS GAME BIRD RATION'
INGREDIENTS PERCENT("a)
Soy Bean Meal 51.15
Ground Corn Meal 36.03
Limestone 5.20
Alfalfa meal 2.50
Monocal 2.33
Soy Oil 1.50
Salt 0.4
L-Lysine 0.28
Poultry Special 0.28
DL Methionine 0.25
Choline Chloride 0.08
Total 100.0
MINERAL AND VITAMIN CONTENT
Calcium 2.60 "/o
Phosphorus 0.85 %
Chloride 0.30 %
Potassium 1.20 6/0
Magnesium 0.15
Sulfur 0.28 "%o
Sodium 0.15 %
Copper 13.00 mg/kg
Iron 44.00 mg/kg
Manganese 105.00 mg/kg
Selenium 0.28 mg/kg
Zinc 85.00 mg/kg
Vitamin A 10.40 iu/g
Vitamin D 3.60 iuig
Vitamin E 2.40 iu/g
Biotin 0.10 mg/kg
Choline 450.00 mg/kg
The guaranteed analysis is a minimum of 27%crude protein,a minimum of 3.7%crude fat and a maximum of 3.5%
crude fiber. Game bird ration supplied by Delmarva Feed and Farm Scrvice,Kennedyvillc,MD.
PROTOCOL NO.: 1004/102423/QLD-SEQ;I00P-409

r
STUDY NO.: 1004B-101
-44-
Appendix II
Diet Formulation
Eurofins Game Bird Ration'
INGREDIENTS PERCENT(%)
Soy Bean Meal 51.15
Ground Corn Meal 36.03
Limestone 5.20
Alfalfa meal 2.50
Monocal 2.33
Soy Oil 1.50
Salt 0.4
L-Lysine 0.28
Poultry Special 0.28
DL Methionine 0.25
Choline Chloride 0.08
Total 100.0
MINERAL AND VITAMIN CONTENT
Calcium 2.60 %
Phosphorus 0.85 %
Chloride 0.30 %
Potassium 1.20 %
Magnesium 0.15 %
Sulfur 0.28 %
Sodium 0.15 %
Copper 13.00 mg/kg
Iron 44.00 mg/kg
Manganese 105.00 mg/kg
Selenium 0.28 mg/kg
Zinc 85.00 mg/kg
Vitamin A 10.40 iu/g
Vitamin D 3.60 iu/g
Vitamin E 2.40 iu/g
Biotin 0.10 mg/kg
Choline 450.00 mg/kg
' The guaranteed analysis is a minimum of 27%crude protein,a minimum of 3.7%crude fat and a maximum of 3.5%
crude fiber. Supplied by Delmarva Feed and Farm Service,Kennedyville,MD

STUDY NO.: 1004B-101
-45-
Appendix Ill
Page 1
Dosing Regime from a Northern Bobwhite Acute Oral
Toxicity Study with Bluegreen Lake Guard Oxy
Experimental Amount of
Group Body Test
Stage (mg a.i./kg) Pen Weight(g) Substance(g)
Control 0 A32 230 NA
0 A33 198 NA
0 A34 231 NA
0 A35 229 NA
0 A36 237 NA
Limit 2000 A37 229 0.550
2000 A38 216 0.519
2000 A39 217 0.521
2000 A40 233 0.559
2000 A41 202 0.485
2 392 A67 214 0.101
497 A68 203 0.121
630 A69 232 0.175
800 A70 200 0.192
1014 A71 223 0.271
1286 A72 203 0.313
1632 A73 209 0.409
2070 A74 202 0.502
2625 A75 221 0.696
3330 A76 238 0.951
NA=Not applicable.Control birds received an empty capsule.
1

STUDY NO.: 1004B-101
-46-
Appendix 111
Page 2
Dosing Regime from a Northern Bobwhite Acute Oral
Toxicity Study with Bluegreen Lake Guard Oxy
Experimental Amount of
Group Body Test
Stage (mg a.i./kg) Pen Weight(g) Substance(g)
3 1281 A97 218 0.335
1281 A98 227 0.349
1626 A99 243 0.474
1626 A 100 222 0.433
2065 A 101 224 0.555
2065 A102 231 0.573
2622 A 103 207 0.652
2622 A104 203 0.639
3330 A105 204 0.816
3330 A106 232 0.927

1
STUDY NO.: 1004B-101
-47—
Appendix IV
Page 1
Daily Observations from a Northern Bobwhite
Acute Oral Toxicity Study with Bluegreen Lake Guard Oxy
Experimental Day of Test
Group 1w 1
(mg a.i./kg) Pen Sex 0816-0821' 0821-0900" 11917° 0945' 1022 1201 1300 1408 AM PM
O A32 M Dosed AN AN AN AN AN AN AN AN AN
0 A33 M Dosed AN AN AN AN AN AN AN AN AN
O A34 M Dosed AN AN AN AN AN AN AN AN AN
O A35 F Dosed AN AN AN AN AN AN AN AN AN
0 A36 M Dosed AN AN AN AN AN AN AN AN AN
2000 A37 M Dosed AN AN AN WF,11 WF,4 11 11,14 II 11
2000 A38 M Dosed AN AN AN AN AN AN AN AN AN
2000 A39 M Dosed AN AN AN AN AN AN SL11 AN AN
2000 A40 F Dosed AN AN WF,4,11 WF,4,11 WF,4 11 11,14 11 11
2000 A41 F Dosed AN 4,13 FD - - - - - -
"-Multiple observations were done follow ing dosing on Day 0 of the test.
'-No regurgitation seen from time of dosing to 1021.
(-)No data available due to mortality.
AN=appeared normal.FD=found dead,SL=slight(used as a modifier).WF=white froth around mouth,
4=loss of coordination.I I =ruffled appearance.13=lower limb weakness. 14=Iethargg

STUDY NO.: 1004B-101
-48—
Appendix IV
Page 2
Daily Observations from a Northern Bobwhite
Acute Oral Toxicity Study with Bluegreen Lake Guard Oxy
Experimental Day of Test
Group 2 3 4 5 6
(mg a.i./kg) Pen Sex AM PM AM PM AM PM AM PM AM PM
O A32 M AN AN AN AN AN AN AN AN AN AN
0 A33 M AN AN AN AN AN AN AN AN AN AN
O A34 M AN AN AN AN AN AN AN AN AN AN
O A35 F AN AN AN AN AN AN AN AN AN AN
O A36 M AN AN AN AN AN AN AN AN AN AN
2000 A37 M 11 AN 11 lI II 11 11,SL(2,4,14) 11,SL(2,4,14) 11,14 11,14
2000 A38 M AN AN AN AN AN AN AN AN AN AN
2000 A39 M AN AN AN AN AN SL11 SLIT 11 AN AN
2000 A40 F AN AN AN AN AN AN 11 11 11,14 11,14
2000 A4I F - - - - - - - - - -
(-)No data available due to mortality.
AN=appeared normal.SL=slight(used as a modifier).2=reduced reaction.4=loss of coordination. 11=ruffled appearance. 14=lethargy

STUDY NO.: 1004B-101
-49—
Appendix IV
Page 3
Daily Observations from a Northern Bobwhite
Acute Oral Toxicity Study with Bluegreen Lake Guard Oxy
Experimental Day of Test
Group 7 8 9 10
(mg a.i./kg) Pen Sex AM PM AM PM AM PM AM PM
0 A32 M AN AN AN AN AN AN AN AN
0 A33 M AN AN AN AN AN AN AN AN
0 A34 M AN AN AN AN AN AN AN AN
0 A35 F AN AN AN AN AN AN AN AN
0 A36 M AN AN AN AN AN AN AN AN
2000 A37 M 11,14 11,14 11,14 11,14 11,14 11,14 11,14 1,2,11
2000 A38 M 11 AN AN AN AN AN AN AN
2000 A39 M 11,14 11 11 11 11,14 11,14 11 AN
2000 A40 F 11,14 11,14 11,14 11,14 4,11,14,EU - - -
2000 A41 F - - - - - - - -
(-)No data available due to mortality.
AN=appeared normal,EU=cuthanized due to very thin condition and weight(1119g),
1 =depression.2=reduced reaction.4=loss of coordination, I I =ruffled appearance. 14=lethargy

zz• z 13
II O No tJ tJ No tJ I " -c (CD
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y I
c c,
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t II ZZ — I>
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tJ > > > > > > > - ,-. :'!
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o y tz N .A X
— " ZZ �— zZZZZ iz M O
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Zzo zzzzz �K ,
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�' ZZ zzzzz 1' 0
5
zz zzzzz K
cn
0
zz ' zzzzz '? z
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0
zz ' zzzzz
W
o I

1
STUDY NO.: 1004B-101
- 51 —
Appendix IV
Page 5
Daily Observations from a Northern Bobwhite
Acute Oral Toxicity Study with Bluegreen Lake Guard Oxy
Experimental Day of Test
Group 0' 1
(mg a.i./kg) Pen Sex 0810-0814" 0814-0945" 1035 1 138 1252 1340 1442 AM PM
392 A67 F Dosed AN AN AN AN AN AN AN AN
497 A68 M Dosed AN AN AN AN AN AN AN AN
630 A69 M Dosed AN AN AN AN AN AN AN AN
800 A70 F Dosed AN AN AN AN AN AN AN AN
1014 A71 F Dosed AN AN AN AN AN AN AN AN
1286 A72 M Dosed AN AN AN AN AN AN AN AN
1632 A73 M Dosed AN AN AN AN AN AN 11 II
2070 A74 F Dosed AN AN AN AN AN AN AN AN
2625 A75 M Dosed AN AN AN AN AN AN 11 Il
3330 A76 M Dosed AN WF WF,4,11,13 FD - - - -
-Multiple observations were done following dosing on Day 0 of the test.
b-No regurgitation seen from time of dosing to 1014.
(-)No data available due to mortality
AN=appeared nonnal.FD=found dead.WF=white froth around mouth.4=loss of coordination.
11 =ruffled appearance. 13=lower limb weakness

-I
STUDY NO.: 1004B-101
-52-
Appendix IV
Page 6
Daily Observations from a Northern Bobwhite
Acute Oral Toxicity Study with Bluegreen Lake Guard Oxy
Experimental Day of Test
Group 2 3 4 5 6
(mg a.i./kg) Pen Sex AM PM AM PM AM PM AM PM AM PM
392 A67 F AN AN AN AN AN AN AN AN AN AN
497 A68 M AN AN AN AN AN AN AN AN AN AN
630 A69 M AN AN AN AN AN AN AN AN AN AN
800 A70 F AN AN AN AN AN AN AN AN AN AN
1014 A71 F AN AN AN AN AN AN AN AN AN AN
1286 A72 M AN AN AN AN AN AN AN AN AN AN
1632 A73 M SL11 11 AN AN AN AN AN AN AN AN
2070 A74 F AN AN AN AN AN AN AN AN AN AN
2625 A75 M SL11 SL11 AN AN AN AN AN AN AN AN
3330 A76 M - - - - - - - - - -
(-)No data available due to mortality
AN=appeared normal.SL=slight(used as a modifier). I I =ruffled appearance

1
STUDY NO.: 1004B-101
-53—
Appendix IV
Page 7
Daily Observations from a Northern Bobwhite
Acute Oral Toxicity Study with Bluegreen Lake Guard Oxy
Experimental Day of Test
Group 7 11 9 10
(mg a.i./kg) Pen Sex AM PM AM PM AM PM AM PM
392 A67 F AN AN AN AN AN AN AN AN
497 A68 M AN AN AN AN AN AN AN AN
630 A69 M AN 11 11 II SLI 1 SLIT AN AN
800 A70 F AN AN AN AN AN AN AN AN
1014 A71 F AN AN AN AN AN AN AN AN
1286 A72 M AN AN AN AN AN AN AN AN
1632 A73 M AN AN AN AN AN AN AN AN
2070 A74 F AN AN AN AN AN AN AN AN
2625 A75 M AN AN AN AN AN AN AN AN
3330 A76 M - - - - - - - -
(-)No data available due to mortality
AN=appeared normal,SL=slight(used as a modifier), 11 =ruffled appearance

.
STUDY NO.. 1004B-101
-54—
Appendix IV
Page 8
Daily Observations from a Northern Bobwhite
Acute Oral Toxicity Study with Bluegreen Lake Guard Oxy
Experimental Day of Test
Group I I 12 13 14
(mg a.i./kg) Pen Sex AM PM AM PM AM PM AM PM
392 A67 F AN AN AN AN AN AN AN AN
497 A68 M AN AN AN AN AN AN AN AN
630 A69 M AN AN AN AN AN AN AN AN
800 A70 F AN AN AN AN AN AN AN AN
1014 A71 F AN AN AN AN AN AN AN AN
1286 A72 M AN AN AN AN AN AN AN AN
1632 A73 M AN AN AN AN AN AN AN AN
2070 A74 F AN AN AN AN AN AN AN AN
2625 A75 M AN AN AN AN AN AN AN AN
3330 A76 M - - - - - - - -
(-)No data available due to mortality
AN=appeared normal

STUDY NO.: 1004B-101
-55—
Appendix IV
Page 9
Daily Observations from a Northern Bobwhite
Acute Oral Toxicity Study with Bluegreen Lake Guard Oxy
Experimental Day of Test
Group 0' 1
(mga.i./kg) Pen Sex 0810-0814" 0922" 1031 1146 1243 1337 1448 AM PM
•
1281 A97 M Dosed AN AN AN AN AN AN AN AN
1281 A98 M Dosed AN AN AN AN AN AN AN AN
1626 A99 M Dosed WF WF,11 11 11 11 11 11 11
1626 A100 F Dosed AN AN AN AN AN AN AN AN
2065 A101 F Dosed WF WF AN AN AN AN AN AN
2065 A102 F Dosed AN AN AN AN AN AN AN AN
2622 A103 M Dosed AN AN 11 11 11 11 11,14 11
2622 A104 M Dosed AN AN 11 11,14 11,14 4,5,11,14 FD -
3330 AI05 M Dosed AN AN AN 11 SL11 SLIT 11,14 11,14
3330 A106 M Dosed AN AN 11 1,5,11 FD - - -
-Multiple observations were done following dosing on Day 0 of the test.
"-No regurgitation seen from time of dosing to 1014.
(-)No data Mailable due to mortality
AN=appeared normal.FD=found dead.SL=slight(used as a modifier).WF=white froth around mouth. I =depression.
4=loss of coordination,5=prostrate posture, 11=ruffled appearance. 14=Ietharny

STUDY NO.: 1004B-101
-56—
Appendix IV
Page 10
Daily Observations from a Northern Bobwhite
Acute Oral Toxicity Study with Bluegreen Lake Guard Oxy
Experimental Day of Test
Group 2 3 4 5 6
(mg a.i./kg) Pen Sex AM PM AM PM AM PM AM PM AM PM
1281 A97 M AN AN AN AN AN AN AN AN AN AN
1281 A98 M AN AN AN AN AN AN AN AN AN AN
1626 A99 M SLII SLIT 11 11 I1 11 SLIT SLI1 11 11
1626 A100 F AN AN AN AN AN AN AN AN AN AN
2065 A101 F AN AN AN AN AN AN AN AN AN AN
2065 A102 F AN AN AN AN AN AN AN AN AN AN
2622 A103 M 11 I 11,14 11,14 II 11,14 11 11 11 11,14
2622 A104 M - - - - - - - - - -
3330 A105 M SLII SL11 SLII SLII AN AN AN AN AN AN
3330 A106 M - - - - - - - - - -
(-)No data m ailable duc to mortality
AN=appeared normal,SL=slight(used as a modifier). 1 I =ruffled appearance. 14=lethargy

1
STUDY NO.: 1004B-101
-57—
Appendix IV
Page 11
Daily Observations from a Northern Bobwhite
• Acute Oral Toxicity Study with Bluegreen Lake Guard Oxy
Experimental Day of Test
Group 7 8 9 10
(mg a.i./kg) Pen Sex AM PM AM PM AM PM AM PM
1281 A97 M AN AN AN AN AN AN AN AN
1281 A98 M AN AN AN AN AN AN AN AN
1626 A99 M 11,14 11,14 11,14 11,14 11,14 11,14 11,14 11
1626 A I00 F AN AN AN AN AN AN AN AN
2065 A101 F AN AN AN AN AN AN AN AN
2065 A102 F AN AN AN AN AN AN AN AN
2622 A103 M FD - - - - - - -
2622 A104 M - - - - - - - -
3330 A105 M AN AN AN AN AN AN AN AN
3330 A106 M - - - - - - - -
(-)No data available due to mortality
AN=appeared normal,FD=found dead, 11=ruffled appearance. 14=lethargy

STUDY NO.: 1004B-101
-58—
Appendix IV
Page 12
Daily Observations from a Northern Bobwhite
Acute Oral Toxicity Study with Bluegreen Lake Guard Oxy
Experimental Day of Test
Group II 12 13 14
(mg a.i./kg) Pen Sex AM PM AM PM AM PM AM PM
1281 A97 M AN AN AN AN AN AN AN AN
1281 A98 M AN AN AN AN AN AN AN AN
1626 A99 M 11 11 11 11 11 11 SL11 SL11
1626 A100 F AN AN AN AN AN AN AN AN
2065 A 101 F AN AN AN AN AN AN AN AN
2065 A102 F AN AN AN AN AN AN AN AN
2622 A103 M - - - - - - - -
2622 A104 M - - - - - - - -
3330 A105 M AN AN AN AN AN AN AN AN
3330 A106 M - - - - - - - -
(-)No data m affable due to mortality
AN=appeared normal.SL=slight(used as a modifier), I I =ruffled appearance

r
STUDY NO.: 1004B-1 Ol
-59-
Appendix V
Page 1
Individual Body Weights(g)from a Northern Bobwhite
Acute Oral Toxicity Study with Bluegreen Lake Guard Oxy
Experimental
Group Total
Stage (m, a.i 1:g) Pen Day-1 Change Day 3 Change Day 7 Change Dav 14 Change
Control () A32 230 -6 224 0 224 6 230 0
i1 A33 198 -I 197 -1 196 5 201 3
o A34 231 -6 225 -6 219 3 222 -9
o A35 229 -10 219 -3 216 10 226 -3
u A36 237 -8 229 1 230 8 238 1
Mean 225 -6 219 -2 217 6 223 -2
SD 15 3 13 3 13 3 14 5
CV(%) 7 6 6 6
Limit 2000 A37 229 -33 196 -28 168 - - -
2000 A38 216 -15 201 -3 198 -7 191 -25
2000 A39 217 -30 187 -18 169 44 213 -4
2000 A40 233 -42 191 -56 135 - - -
2000 A41 202 - - - - - - -
Mean 219 -30 194 -26 168 19 202 -15
SD 12 II 6 22 26 36 16 15
CV(%) 6 3 15 8
2 392 A67 214 -16 198 -3 195 11 206 -8
497 A68 203 -12 191 I 192 9 201 -2
630 A69 232 -25 207 -23 184 26 210 -22
800 A70 200 -16 184 1 185 10 195 -5
1014 A 7I 223 -21 202 -7 195 12 207 -16
1286 A72 203 -23 180 3 183 11 194 -9
1632 A73 209 -20 189 3 192 14 206 -3
2070 A74 202 -26 176 -7 169 24 193 -9
2625 A75 221 -18 203 0 203 14 217 -4
3330 A76 238 - - - - - - -
(-)No data available due to mortality.
SD=standard deviation,CV=coefficient of variation

n
STUDY NO.: 1004E-101
-60-
Appendix V
Page 2
Individual Body Weights(g)from a Northern Bobwhite
Acute Oral Toxicity Study with Bluegreen Lake Guard Oxy
Experimental
Group Total
Stage (mg a.i./kg) Pen Day-1 Change Day 3 Change Day 7 Change Day 14 Change
3 1281 A97 218 -12 206 4 210 11 221 3
1281 A98 227 -26 201 3 204 15 219 -8
Mean 223 -19 204 4 207 13 220 -3
SD 6 10 4 1 4 3 1 8
CV(%) 3 2 2 1
1626 A99 243 -30 213 -22 191 -13 178 -65 1
1626 A100 222 -23 199 0 199 13 212 -10
Mean 233 -27 206 -11 195 0 195 -38
SD 15 5 10 16 6 18 24 39
CV(%) 6 5 3 12
2065 A101 224 -30 194 -20 174 13 187 -37
2065 A102 231 -21 210 6 216 8 224 -7
Mean 228 -26 202 -7 195 11 206 -22
SD 5 6 11 18 30 4 26 21
CV(%) 2 6 15 13
2622 A103 207 -47 160 - - - - -
2622 A104 203 - - - - - - -
Mean 205 -47 160 - - - - -
SD 3 - - - - - - -
CV(%) 1 - - -
3330 A105 204 -29 175 1 176 14 190 -14
3330 A106 232 - - - - - - -
Mean 218 -29 175 1 176 14 190 -14
SD 20 - - - - - - -
CV(%) 9 - - -
(-)No data available due to mortality.
SD=standard deviation,CV=coefficient of variation

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STUDY NO.: 1004E-101
-62-
Appendix VI
Page 2
Individual Feed Consumption(g/bird/day)from a Northern Bobwhite
Acute Oral Toxicity Study with Bluegreen Lake Guard Oxy
Estimated Mean Feed
Experimental Consumption
Pe
Group Grams/Bird/Day
Stage (mg a.i./kg) Pen Day 0-1 Day 1-2 Day 2-3 Day 3-7 Day 7-14
3 1281 A97 6 13 11 12 13
1281 A98 6 6 5 8 15
Mean 6 10 8 10 14
SD 0 5 4 3 1
1626 A99 4 4 4 1 5
1626 A100 5 6 6 10 15
Mean 5 5 5 5 10
SD 1 1 1 7 7
i
2065 AIOI 5 4 4 I 8
2065 A102 5 9 0 15 15
Mean 5 7 2 8 12
SD 0 4 3 10 5
2622 A103 5 5 1 - -
2622 A104 - - - - -
Mean 5 5 I - -
SD - - - - -
3330 A105 4 6 16 6 13
3330 A106 - - - - -
Mean 4 6 16 6 13
SD - - - - -
(-)No data available due to mortality.
SD=standard deviation.

r
STUDY NO.: 1004E-101
-63 -
Appendix VII
Personnel Involved in the Study
The following key personnel were involved in the conduct or management of this study:
(I) Diana L. Temple, M.S.
(2) Patrick M. Hubbard,B.S.
(3) Ryan J. Davis

TO:
BlueGreen DATE: Richard Rogers
WATER TECHNOLOGIES March 21,2023 Director
North Carolina
Department of Environmental Quality
March 21,2023
Via Electronic Mail
Re: BlueGreen US Water Technologies, LLC—Response to Technical Group Questions
Concerning Lake Guard® Oxy
Please find below BlueGreen's responses to the technical group's questions.
1. Full range Ceriodaphnia dubia(native species)chronic test using EPA Method following
EPA Method EPA-821-R-02-013.
Response: BlueGreen has selected Stillmeadow, Inc. to conduct the chronic toxicity testing
on Ceriodaphnia duhia following EPA Method EPA-821-R-02-013. Stillmeadow, Inc. is
AAALAC-accredited and certified with the National Environmental Laboratory Accreditation
Program(NELAP). Stillmeadow,Inc.is also certified by the Japanese Ministry of Agriculture,
Forestry and Fisheries(MAFF)in 1992.
2. Once a safe dosage for aquatic life is determined,will BlueGreen reassess minimum and
maximum amounts to be applied and can anticipated application rates be provided in
volumetric measurements(mg/L) to compare with toxicity information more easily?
Also, since application amounts may vary,describe evaluation,decision,and
notification process that will occur prior to treatment.
Response: Once a safe dosage for aquatic life is determined through chronic toxicity testing,
BlueGreen will reassess minimum and maximum amounts to be applied. A follow-up
discussion between stakeholders will need to address what formal decisions will be made
should mortality of Ceriodaphnia dubia be caused by Lake water alone(control group)without
the addition of any Lake Guard®Oxy(treatment group).
Dosage of treatment application will also strictly adhere to directions provided on the EPA
approved certified by NSF/ANSI/CAN-60 product label. The product label provides:
"If there is some doubt about the amount of Lake Guardr''Oxy to apply, it is
best to start with the lower application rate. Since all water bodies are unique
aquatic ecosystems and, therefore, differ from each other in treatment
response and longevity, the Lake Guard''Oxy application protocol offers the
applicator a scalable dose rate that saves money and reduces the chemical
load introduced to the environment. For first time users, or when in doubt,
applicators should start with 0.5-2.5 lbs acre and adjust the dose rates
(increase or decrease) by increments of 25-50%, and over time can,fine-tune
the treatment application for each waterhody."
CONTACT: 11b1..11O
.11,
1-866-269-2828 [e-mail] 301 South Hills Village Ste
www.bluegreenwatertech.com LL200#452,Pittsburgh,PA
15241

L

TO:
BlueGreen DATE: Richard Rogers
WATER TECHNOLOGIES March 21,2023 Director
North Carolina
Department of Environmental Quality
After considering the chronic toxicity testing results, dosage will be dependent upon the
concentration of cyanobacterial cells per volume of water (cell density) present at the time.
Application rate instnictions were taken from the Lake Guard® Oxy product label, which is
summarized in the table below. All rates in lbs/acre have been converted to mg/L. Again,
these application rates may be adjusted depending on the chronic toxicity testing results.
BlueGreen has also created a decision tree diagram (see below), which is for illustration
purposes only,provisional,and subject to change. The decision tree will be revised to include
consideration of the chronic toxicity testing results. The decision tree uses quantitative metrics
to make informed decisions to produce more favorable outcomes. Furthermore, the decision
tree diagram serves as a guide using steps to determine dosing amounts under various
cyanobacterial cell densities, chlorophyll-a concentrations, and cyanotoxins' concentrations.
These specific values were taken from World Health Organization. 2003. Guidelines for safe
recreational water environments, Volume 1, coastal and fresh waters (Guidelines on
recreational water quality: Volume 1 coastal and fresh waters(who.int)). While no regulation
exists at the federal and state level,EPA and many states recognize the WHO 2003 guidelines
and use its guiding principles to aid in human health risks to various forms of water pollution.
Lake Guard® OXY Dosage Instructions
Cyanobacteria cell density Cyanobacteria biomass Dose Amount Dose Conversion:(lbs/acre)to(mg/L)
(cells/mi) (Chia pg/L.) (lbs/acre) Dose Amount(mg/L or ppm)
5000-20000 10 0.5-5 0.18.1.85
20000-100000 to •,o ;-i0 1.85-1 1.10
>100000 >50 30-98 11.10-36.26
>98 46.26 ...
11111111111111111111111110111111eY (b_ <. wa( dy area)
294 Not to exceed 108.11
(MAXIMUM single application rate allowed)
CONTACT:
1-866-269-2828 [e-mail] 301 South Hills Village Ste
www.bluegreenwatertech.com LL200#452,Pittsburgh,PA
15241

I
I

TO:
0 B I u e G ree n DATE: Richard Rogers
W A T E R TECHNOLOGIES March 21,2023 Director
North Carolina
Department of Environmental Quality
DOSING DECISION TREE
LOW LEVEL "FREE" AMOUNT OF DOSING WILL STRICTLY ADHERE TO THE GUIDELINES
Cell count LOW DOSE Cell count YES PROVIDED BY THE LAKE GUARD OXY PRODUCT LABEL
5,000-20,000 cells/ml !!. <5,000 cells/mL __.�� NO DOSE FOR ILLUSTRATION PURPOSES ONLY
/
Microcystin-LR Microcystin LR PROVISIONAL AND SUBJECT TO CHANGE
<10 µg/L ND µg/L
Chlorophyl-a
<10 µg/L "FREE"
Cell count YES NO DOSE
YES <5,000 cells/mL �e"
MODERATE LEVEL LOW LEVEL —► Microcystin-LR
"FREE"
START Cell count Cell count ND µg/L LOW LEVEL Cell count
Cyanobacteria/ '2*-- 20,000 100,000 cells ml MODERATE DOSE YES
/ 5,000 20,000 cells/mL Cell count YES <5,000 cells/ml
Cyanotoxin Microcystin-LR ��� NO DOSE
Levels �� Microcystin-LR 5,00020,000 cells/ml Microcystin-LR
10-20 µg/L 40 µg/L NOr LOW DOSE Microcystin LR ND µg/L
Chlorophvll-a <10 µg/L
10-50 µg/L
"FREE"
Cell count
LOW LEVEL <5,000 cells/mL YES NO DOSE
Cell count LOW DOSE Microcystin-LR ��
HIGH LEVEL MODERATE LEVEL YES 5,000 20,000 cells/mL �� ND µg/L
', Cell count Cell count --O. Microcystin LR
100,000-10,000,000 cells/mi. HIGH DOSE 20,000-100,000 cells/mL <10 µg/L LOW LEVEL
Microcvstin-LR
Microcystin-LR
• 20 2,000 µg/I Cell count
10 20 µg/L NO YES
Chlorophyll-a �► MODERNE DOSE 5,000-20,000 cells/mt. �� NO DOSE
50 5,000 µg/L Microcystin-lR
<10 µg/l
MODERATE LEVEL "FREE"
VERY HIGH LEVEL YES LOW LEVEL
HIGH LEVEL Cell count YES
Cell count HIGHER Cell count ""'.."� Cell count Cell count
20,000-100,000 cells/mL MODERATE DOSE LOW DOSE -�'i'
>10,000,000 cells/mL DOSE 5,000-20,000 cells/ml <5,000 cells/mt. NO DOSE
100,00610,000,000 cells/mL Nlicrocvstin LR �'�
Microcystin-LR * Microcvstin-LR Microcystin-IR �"�' Microcystin-LR
>2,000 µg/l 10-20 µg/L <10
20-2,000 µg/L µB/l ND µg/L
Chlorophyll-a NO
>5,000 µg/L
HIGH DOSE 11/
CONTACT: %N. , r„ O
1-866-269-2828 [e-mail] 301 South Hills Village Ste
www.bluegreenwatertech.com 1 LL200#452,Pittsburgh,PA
15241

TO:
DATE: Richard Rogers
6 BlueGreen March 21.2023 Director
WATER TECHNOLOGIES North Carolina
Department of Environmental Quality
3. The Lake Guard Oxy product label states, "Do not apply to shallow water bodies less
than three feet deep". Presumably, almost all sites that have been treated contain
shoreline areas less than three feet deep. Please elaborate on the intent of this statement.
Response: The product label's recommendation to not apply the product to "shallow water
bodies less than three feet deep" was specifically requested by BlueGreen for marketing
u ses to differentiate between the consumer-grade versus professional-grade category.
P � �' P �'
There is no scientific basis to prohibit using the product in water bodies less than three feet
deep. This depth limitation is not necessary to protect public health, wildlife, aquatic
resources, or the environment. The product should not be applied haphazardly and should be
applied by a licensed applicator.
On January 30, 2023, BlueGreen submitted the Notification for Lake Guard® Oxy to EPA
requesting that this recommendation be deleted from the label as follows:
OLD: "Do not apply to shallow water bodies less than three feet deep,or to areas where surface
water is stagnant. Do not use the product to treat ornamental fish (i.e.,fish that grow in home
aquariums or outdoor tanks for aesthetic purposes)."
NEW:"
water Do not use the product to treat ornamental fish (i.e., fish that grow in home
aquariums or outdoor tanks for aesthetic purposes)."
BlueGreen's modification request was approved by the EPA on March 9,2023.
4. Please elaborate on the toxicity to birds which is indicated on the label. tinder what
conditions was it toxic (ingestion only)? Was it only toxic at certain application rates?
Were certain species or taxa more vulnerable?
Response: Sodium percarbonate("SPC")is a solid compound that,when dissolved in water,
releases hydrogen peroxide and soda ash (sodium carbonate). The U.S. National Organic
Standards Board concluded that "SPC is one of the most desirable compounds to use as a
cyanobacterial algaecide in food-fish ponds due to its environmentally-safe nature i.e.,no toxic
residuals are formed from the breakdown of SPC." A more detailed excerpt is provided below:
"SPC rapidly dissociates into hydrogen peroxide and sodium carbonate.
Hydrogen peroxide is the active component. Hydrogen-peroxide-based
products work by exogenously oxidizing organic matter. However, because
hydrogen peroxide is a natural byproduct of cell metabolism, present at a low
level in all living cells,and secreted by some bacteria,living organisms are
naturally subject to oxidative damage from both exogenous and endogenous
hydrogen peroxide(Apel and Hirt,2004). In their defense,living cells are
CONTACT: 1 1.
1-866-269-2828 [e-mail] 301 South Hills Village Ste
www.bluegreenwatertech.com LL200#452,Pittsburgh,PA
15241

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TO:
DATE: Richard Rogers
6 BlueGreen March 21.2023 Director
WATER TECHNOLOGIES North Carolina
Department of Environmental Quality
naturally protected from endogenous hydrogen peroxide by an antioxidant
protection system composed of three enzymes: superoxide dismutase, catalase
and glutathione peroxidase. Moreover two vitamins, vitamin C and vitamin E
also serve as intracellular antioxidant scavengers(MDEP, 2010). SPC is one of
the most desirable compounds to use as a cyanobacterial algaecide in food-fish
ponds due to its environmentally-safe nature i.e.,no toxic residuals are formed
from the breakdown of SPC. in its use for the purpose of disinfecting catfish
eggs, SPC was found to be efficacious, and contributed to the beneficial
increase of dissolved oxygen in the hatching trough (Small, 2009). The
breakdown products of SPC are water, oxygen and sodium carbonate,none of
which concentrate or persist in the environment or represent environmental
hazards."
The metabolism of animals includes the production of hydrogen peroxide through enzymatic
processes as a natural defense mechanism. Veterinarians will prescribe administering
hydrogen peroxide to dogs to induce vomiting from consuming contaminants jeopardizing the
health of the dog. Up to 30% concentration of hydrogen peroxide is considered safe, while
40% concentration is considered unsafe. Percent concentration of Lake Guard® Oxy from
BlueGreen is in the range of 27-30% of hydrogen peroxide. In addition, due to the nature of
the time-releasing Lake Guard®Oxy formula, the actual dosage of hydrogen peroxide that is
present in the water is a few orders of magnitude than the theoretical concentration of 27-30%,
as indicated at the model below in reference to Question#6.
Please also refer to the attached documents: Sodium Carbonate Peroxyhydrate TR 2014 and
MassachusettsDEP_Sodium carbonate peroxyhydrate& hydrogen peroxide_2010.
5. Given that the chemical is applied as a floating pellet what is the likelihood that it could
be perceived as food and consumed by other aquatic organisms, such as fish, mammals,
or reptiles?
Response: BlueGreen has never witnessed or experienced aquatic organisms consuming Lake
Guard® Oxy during trials or during thousands of commercial applications. On the contrary,
results within the Lake Guard® Oxy Efficacy Report March 2020 showed that the fish in the
study did not come close to the floating particles. In no case were fish seen nibling on or
otherwise being attracted to the floating product or affected by it. In addition, a safety trial
with the Israeli Ministry of Agriculture during 2022 also indicated that Lake Guard®Oxy was
safe for fish (a report in Hebrew from the Experimental Farm at Dor, Israel, is attached with
its translation to English).
One of two main reasons for that are that the dissolution of the Lake Guard® Formula to the
water is slow (as indicated in the model below in Question #6), and thereby the actual H202
concentration in the water at any moment is in the range of 10-6-10-7 ppm. Secondly,the fish
CONTACT: I �:=� 00
1-866-269-2828 [e-mail] 301 South Hills Village Ste
www.bluegreenwatertech.com 1.1200#452,Pittsburgh,PA
15241

1

TO:
DATE: Richard Rogers
6 BlueGreen March 21,2023 Director
WATER TECHNOLOGIES North Carolina
Department of Environmental Quality
mouth cavity contains nociceptors-a sensory receptor for painful stimuli and detects chemicals
in the water-which explains why they do not approach the white pallets and nibble upon them.
6. What is the release rate of the encapsulated Lake Guard Oxy?
Response: Lake Guard'Oxy(LGO)is a time-releasing active ingredient(H202)encapsulated
by an inert and biodegradable hydrophobic shell that floats on the water surface, migrating
together along the same wind/current direction as cyanobacteria. The formulation contains 2%
coating and 27%H202 and the degradation rate will be dependent on the turbidity(k)of water.
Turbidity is caused by particles suspended or dissolved in water that scatter light making the
water appear cloudy or murky. Particulate matter can include sediment - especially clay and
silt, fine organic and inorganic matter, soluble colored organic compounds, algae, and other
microscopic organisms. Turbidity can range from 0-1 NTU, where higher values represent
higher particulate matter in the water. Here BlueGreen presents the degradation rate of various
doses of LGO ranging from 10-500 pounds per acre-ft over time from 0 to 60 minutes and
under turbidity conditions of 0.1,0.5 and 0.9 NTU. Notably,the maximal dosage allowed by
the label for LGO is 298 lb/acre-ft, nevertheless, here BlueGreen used a 500 lb/acre-ft as an
overdose.
The equation for the rate of H202 degradation is:
Ct = Co * exp (—kt)
Where:
C,is the concentration at time t.
Co is the initial peroxide concentration
k is the turbidity coefficient
t is the time interval
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TO:
DATE: Richard Rogers
6 BlueGreen March21.2023 Director
WATER TECHNOLOGIES North Carolina
Department of Environmental Quality
se-06-
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'_GO(Pounds per acre fti
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TO:
A B I lJ P.Green DATE: Richard Rogers
March 21,2023 Director
WATER TECHNOLOGIES North Carolina
Department of Environmental Quality
C
a
LGO(Pounds per acre ft)
d _
x 10
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100
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a, 500
o`
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Time after treatment(min)
CONTACT:
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15241

TO:
DATE: Richard Rogers
B I u e G ree n March 21,2023 Director
WATER TECHNOLOGIES North Carolina
Department of Environmental Quality
122F-11-
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a LGO(Pounds per acre ft)
- 10
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Time after treatment(min)
As indicated in the graphics above, which does not consider the fact that hydrogen peroxide
dissociates faster under warmer temperatures, under more turbid conditions (i.e., k4,1.5 and k=0.9)
Lake Guard®Oxy dissolves in as little as 30 minutes and as much as 40 minutes,respectively. Under
less turbid conditions (i.e., k=0.1), Lake Guard® Oxy can take up to four hours to dissolve. It is
important to bear in mind that other"foreign"constituents in the water column,beyond BlueGreen's
control, can affect any chemical reaction. The rate of decomposition of H202 in its purest form
increases with rise in temperature, concentration, and pH, meaning H202 is unstable under alkaline
conditions. However, H202 shows the best stability with cool,dilute,and acidic solutions.
7. The efficacy table found in the Lake Guard Oxy Efficacy Report provided biomass and
percent biomass reductions for only cyanobacteria,were other alga present? If so, does
similar data exist?
Response: The focus of the Lake Guard Oxy Efficacy Report was only on cyanobacterial
biomass present. Lake Guard® Oxy, however, will also treat other algae with similar
characteristics. Former treatments using Lake Guard®Oxy have demonstrated the reduction
in cyanobacteria, opening a niche for other beneficial phytoplankton to fill that niche and
dominate.
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.;
,.
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15241

TO:
DATE: Richard Rogers
6 BlueGreen March 21,2023 Director
WATER TECHNOLOGIES North Carolina
Department of Environmental Quality
8. There are several questions about The Environmental Effect of the Use of Lake Guard
Oxy on Fish Population report:
a. Why was the decision made to artificially aerate a system intended to evaluate the
environmental effects of the use of Lake Guard on fish?
Response: All ponds were continuously aerated with a single exemption of one hour,
immediately after applying the Lake Guard® Oxy on the water surface. Fish health-
evaluation was conducted during the first hour after treatment when there was no aeration
and at the end of the trial by a scientist from the Central Laboratory of Fish Health.
b. Were all endpoints of this study made by visual observation?
Response: Yes.
c. Was histology performed on gill tissues to examine for oxidative damage?
Response: No. Fish were weighed and their behavior was observed over days.
Effect on Fish Populations
Despite this reference, post-treatment results using Lake Guard® Oxy in Lake Minneola,
Florida indicated that following the bloom treatment over a period of time of seven months,
there was an increase in biodiversity of fish according to FWC (see attached report).
9. Will there be any interim evaluation metrics and decision points?
Response: BlueGreen will rely on the chronic toxicity testing results, product label
instructions, and the decision tree for application rates and monitoring of concentration of
algal/cyanobacteria and for benchmarks. In addition, BlueGreen will be using in-water probe
devices that provide a continuous flow of data and relevant parameters,which can be remotely
accessed from anywhere in the world with WiFi connection to view a dashboard to view results
in near real-time. The in-water probe devices allow one to set up threshold alerts should any
parameter rise above a desired value. Lastly, BlueGreen will involve the stakeholders in
decisions and directions to be taken for the duration of the research project to determine best
management practices.
10. In previous treatments worldwide, have you ever observed any animal mortality that
could have occurred because of treatment?
Response: No. BlueGreen has never observed any animal mortality that could have occurred
as a result of treatment. There were no reported or observed mortalities in any of the
following former treatments using Lake Guard®Oxy: Lake Minneola Florida;a bird-
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API
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TO:
DATE: Richard Rogers
6 BlueGreen March 21.2023 Director
WATER TECHNOLOGIES North Carolina
Department of Environmental Quality
infested Zoo in Israel; drinking water reservoir in Setumo Dam, South Africa; Pahokee
Marina, Lake Okeechobee, Florida; C-43 and C-44 canals in Caloosahatchee and St. Lucie
Rivers,respectively; Roodeplaat Dam in South Africa; and Lake Anna, Virgina. In fact, as
measured by third parties in Lake Minneola and Setumo Dam, both experienced a positive
feedback loop because of treatment using Lake Guard® Oxy resulting in increases in
biodiversity and sizes of fish.
11. Is there any monitoring for negative effects planned during and following the treatment?
Response: Computationally, the in-water probe devices provide continuous measurements
that have a built-in alert set for specified thresholds, which will be constantly tracked by
BlueGreen. Visual assessments will rely on accounts from the staff, BlueGreen's
subcontractor conducting maintenance of in-water devices, site visits by the project manager
(Dr. Jessica Frost),and periodic flyovers by BlueGreen.
12. At Florida sites which have been treated previously,how long did improvements last,and
did the treatment have to be repeated?
Response: Each water body is unique,and there is no"one-size-fits-all"when managing poor
water quality. Therefore,results from previously treated areas in Florida cannot be correlated
to results that will be achieved at another water body. No negative effects occurred during or
after treatment at the Florida sites,and treated waters were improved across a few months to a
couple years. The Florida treatment sites, however, involved a one-time treatment and there
was not harmful algal bloom ("NAB")treatment plan in place that included subsequent spot
treatments to mitigate HABs.
Each water body has its own unique characteristics,much of which can affect the effectiveness
of an algaecide treatment or any kind of management practice. By performing due diligence
tasks, such as analyzing available historical data, analyzing trends in the water quality data,
gaining knowledge and understanding of biological processes within the ecosystem,including
the number of natural resources available for pelagic,epifaunal and infaunal communities,and
accounting for external factors that contribute to the quality of water, one can make better
informed decisions regarding management, which should include preventative measures.
Below and attached are a few examples of treatments using Lake Guard®Oxy in Florida.
*See attachment"Lake_Minneola-Algal-BloomRemediation-Apr202I.pdf"(SJRWMD)
Click following links:
SFWMD uses innovative technology to combat algae in Caloosahatchee River I South
Central Florida Life
Florida spent SI million on toxic algae in Caloosahatchee, Pahokee Marina(tcpalm.com)
CONTACT: %= 00
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I

TO:
DATE: Richard Rogers
BlueGreen March 21.2023 Director
WATER TECHNOLOGIES North Carolina
Department of Environmental Quality
13. Please provide examples of other localities where this approach has been applied and
been successful.
Response: The attached document, 12-2022_LACA_CMP Final_Report_Full_Version,
provides examples of successful treatment applications. In addition,please see Lake Anna
Civic Association - 2022 CMP Final Report (lakeannavirginia.org).
Mr. Harry Looney of LACA gave permission to share the above and offered:
"You are also welcome to share my contact info if they would like to reach out to me with
any questions. -Harry"
Harry Looney
Water Quality Project Officer,Lake Anna Civic Association
[phone]
www.lakeannavirginia.org
14. Are there other bodies of water in federal ownership where this chemical or precursor
chemicals(e.g.Pak-27)have been used?
Response: The short answer is no. However,the United States Army Corp of Engineers were
aware of treatment activities in and around Lake Okeechobee,Florida since USACE is the sole
operator of opening and closing gate structures that connect Lake Okeechobee to the
Caloosahatchee River via the C-43 canal and the St. Lucie River via the C-44 canal.BlueGreen
can only speak to and has tracking knowledge of Lake Guard® Oxy usage and not Pak-27,
which is widely distributed and applied throughout the United States by various applicators.
15. Will the in-water probes and sampling methodology comply with state-certified methods
established by 40 C.F.R. Part 136?
Response: The in-water probes measure the following:
Sensor Measurements
Parameter Range Resolution
0-50°C 0.1°C
0-50°C 0.1°C
0-200 NTU 0.1 NTU
0-200 ug/L or 0-2,000 RFU 0.1 pg/L or 0.1 RFU
0-1.500 pg/L or 0-750 RFU 0.1 Ng/L or 0.1 RFU
0-2.000 PAR 0.1 pE/m'/sec
All 2.5 m
Code of Federal Regulations Title 40 Chapter 1 Subchapter D Part 136 was drafted primarily
for industrial and municipal water. Since the EPA does not have a recognized Water Program
specific to surface water, states often adopt methods from Part 141 (applicable to potable
water) and Part 136 (applicable to non-potable water) for their surface water regulatory
program.
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1

TO:
DATE: Richard Rogers
6 BlueGreen March 21,2023 Director
WATER TECHNOLOGIES North Carolina
Department of Environmental Quality
In 40 C.F.R Part 136, Dr. Jessica Frost thoroughly searched for EPA requirements regarding
field parameters or field instruments for EPA Region 4,under which North Carolina is included
and found that under subsection 136.3 (see below) Table IB only addressed Turbidity (item
#73)out of the listed metrics in the above sensor measurements table. No other EPA approved
methods were found for field instruments measuring Chlorophyll-a, Phycocyanin or Ambient
Light. Additionally, there is no SOP reference for calibration of field instruments in EPA
Region 4.
"§ 136.3 Identification of test procedures.
(a) Parameters or pollutants, for which methods are approved, are listed together
with test procedure descriptions and references in Tables IA,IB,IC,ID,IE,IF,IG,and
IH of this section."
To that end and because the in-water probes collect only field parameters and do not collect
water samples for in-lab measurements in which EPA/APHA Standard Methods/AOAC,
ASTM,or USGS methodology would be applicable,BlueGreen does,however,abide by EPA
EQASOP-FieldCalibrat3 for EPA Region 1 Calibration of Field Instruments, Revision
Number: 3,Date: June 3, 1998, Revised March 23, 2017 (see""excerpt below).
``Instruments(e.g.,sonde)that monitor continuously over a period of time are calibrated
before deployment. When these instruments are recovered, the calibration is checked
to determine if any of them drifted out of calibration."
It is noteworthy to mention that Dr. Jessica Frost has certificates of TNI training from 2021,
was a chemist for the NELAP/NELAC certified lab of Microbac Laboratories,Inc.conducting
sample analysis of drinking water and wastewater.
Sincerely,
X ��.dt.
Jessica Frost
Scientific Director USA
Jessica Frost, Ph.D
BlueGreen US Water Technologies, LLC
Enclosure
cc: Mike Pelz
Hans Pearl,Ph.D
Nathan Hall, Ph.D
Kendall Smith, USFWS
End Document
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