UNITED STATES ENVIRONMENT AL PROTECTION AGENCY 

WASHINGTON, D.C. 20460 

OCT 1 5 2015 

OFFICE OF CHEMICAL SAFETY 

ANO POLLUTION PREVENTION 

Stephen L. Dobson, Ph.D. 

Uni versity of Kentucky, Department of Entomology 

S-225 Agricultura l Science Center North 

Lexington, KY 40546-0091 

Subject: Experimental Use Pennit Issuance for Wolbachia pipientis, wAlbB Strain in Male Aedes 

aegypti WB l Strain Mosquitoes (Including Egg, Larva l, Pupal, and Adult Life Stages) in 

California 

Experimental Use Permit No.: 88877-EUP-2 

OPP Decision No.: 500458 

Effective Dates: Immediately until December 3 1. 2016 (pesticide applications and 

associated activities, e.g., collection of field data) 

Quantity Authorized: 2.400,000 male Aedes aegypti WBI Strain mosquitoes, weighing 

5.672 ounces (160.8 grams) and containing 5.672 x 10·

5 ounce (l.608 x 10·3 gran1) of 

active ingredient 

Acr es Involved: 840 acres 

Dear Dr. Dobson: 

On the basis of the information furn ished by you, the subject Experimental Use Penn it (EUP) under 

sectio n 5 of the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) is hereby issued for the 

pesticidal active ingredient Wolbachia pipientis, wAlbB Strain in ma le Aedes aegypti WB 1 Strain 

mosquitoes. The EUP, which is authorized only in the state of Cali fornia, will evaluate the active 

ingredient's effectiveness in suppressing and eliminating Aedes aegypti mosquitoes at particular sites in 

Fresno County. Shipment and/or use under this permit are subject to the provisions of 40 CFR Part 172. 

Prior to shipment and/or use of this material. you must consult with the pesticide regulatory officials of 

the states in which you will conduct your experimental program and obtain a state permit or license if 

such is required. Issuance of this Federal permit does not negate the need for pem1ission from individual 

states. Prior to initiating this experimental program in any state. you are required to notify the lead 

agency of the states in which you will conduct your experimental program of the specific testing 

program (when. where. how much, etc.). Failure to obtain that permission may result in revocation or 

modification of the EUP. 

You must provide notice to the state pesticide regulatory agency at least 72 hours prior to the application 

of this experimental pesticide product. You must also provide a copy of this authorization letter and the 
Page 2of3 

EPA EUP No. 88877-EUP-2 

OPP Decision No. 500458 

stamped, accepted EUP labeling to all cooperators, participants, and users prior to the initial pesticide 

application made in accordance with this EUP. The U.S. Environmental Protection Agency (EPA) will 

notify the relevant EPA Region of the approval of this EUP by sending copies of this EUP authori zation 

letter and the stamped, accepted EUP labelfng to·it. 

Based on the experimental program submitted, this product may be shipped for use under this EUP to 

the state of California for the next experimental period, effective immediately until December 31. 2016. 

Within 840 acres of Fresno County. you may release. monitor, and collect data on 2,400.000 male Aedes 

aegypti WB I Strain mosquitoes that weigh 5.672 ounces (oz) ( 160.8 grams (g)) and contain 5.672 x 10·5 

oz (1 .608 x I 0·

3 g) of Wolbachia pipientis, wAlbB Strain. 

You will immediately notify the EPA of any findings from the experimental uses that have a bearing on 

safety (i.e .. the EPA requires reporting of any adverse effects from the use of or exposure to pesticides). 

You will also keep records of production, distribution. and performance, and make the records available 

on request to any authorized officer or employee of the EPA. 

Prior to registration or amendment under FIFRA section 3, all data requirements must be satisfied (40 

CFR Part 158, Subpart V). 

The labeling submitted in connection with the application for the EUP is acceptable. and a stamped. 

accepted copy is enclosed for your records. This labeling must be used for all shipments under this EUP 

and must be in possession of the user at the time of pesticide appli cation. 

You must provide a final report at the conclusion of the experiment. This final report shall include all of 

the items set forth in 40 CFR § I 72.8(b)(2). In addition, you must perform the following testing and 

monitoring and include the results of this testing and monitoring in the final report: 

(1) Sample the mosquito population used to produce male wAlbB Strain Aedes aegypti mosquitoes 

for possible infection with pathogenic viruses such as dengue; Eastern equine encephalitis; West 

Nile virus; other arboviruses; and lymphatic, subcutaneous, and serious cavity fi lariasis. 

(2) Monitor the release of wAlbB Strain-infected Aedes aegypri female mosquitoes at the release 

sites. If polymerase chain reaction (PCR) is used to monitor for the wAlbB Strain, describe how 

these wAlbB Strain-infected female mosquitoes will be distinguished from the wild Aedes 

aegypti female mosquitoes that have mated with wAlbB Strain-infected Aedes aegypti male 

mosquitoes. In other words, describe how you ensure that the wild type female mosquitoes do 

not appear wAlbB Strain positive (using PCR) after fertilization with wAlbB Strain-infected 

male mosquitoes. 

(3) Sample released mosquitoes to confirm the rate of female mosquitoes released. 

( 4) Monitor population density for an impact of male mosquito releases through trapping of Aedes 

aegypti mosquitoes and additional mosquitoes that co-occur at the sites and that are collected in 

traps. 
Page 3of 3 

EPA EUP No. 88877-EUP-2 

OPP Decision No. 500458 

(5) Assess re lease male mosquito quality by holding a subset in a cage and monitoring 

survival/ft tness. 

(6) Mark a subset of release male mosquitoes with dust and then monitor for their recapture (markrelease-recapture). (This addresses questions of the release to indigenous male mosquito ratio 

and release male mosquito survival.) 

(7) Collect eggs from the field to examine for a reduction in egg hatch. 

(8) Monitor environmental conditions (temperature. wind speed, etc.) from a local National Oceanic 

and Atmospheric Administration station. 

If you have any questions regarding this permit. please contact Jeannine Kausch by phone at (703) 347-

8920 or via email at kausch.jeannine@epa.gov. 

Enc losure 

s~Zi~)· lf' 

Robert Mc ally. Director 

Biopesticides and Pollution 

Prevention Division (75 11 P) 

Office of Pesticide Programs 

cc: Patti TenBrook. EP J\ Region 9 (California)