DILUTION of ANVIL 10+10^Ò for the BOTTLE BIOASSAY

DRAFT DOCUMENT FOR REVIEW

PHEREC Technical Memorandum # 1

During September 2001, three Florida Mosquito Control Programs requested assistance in evaluating ANVIL 10+10^Ò as a mosquito adulticide. Workshops were conducted focusing on carrying out serial dilutions of ANVIL 10+10^Ò to determine the diagnostic concentration to be applied to the interior surface of each 250 ml Wheaton bottle following the CDC protocol as modified by Dukes and Greer that is posted to the PHEREC Web site http://pherec.org/bottleassay/bottlebioassay.html

This brief technical memorandum reports the results of those workshops.

IMPORTANT MESSAGE: These technical notes are intended as a provisional report of on-going collaborative work. The conclusions contained in this memorandum may be modified in the future, based on additional laboratory or field experiences. Please go to the PHEREC Web site for updates http://pherec.org Direct inquiries and/or comments to: Dr. Jack Petersen, drjack3@hotmail.com

OBJECTIVE: Determine the diagnostic dose of ANVIL 10+10^Ò for the bottle bioassay.

START with the LABEL. ANVIL 10+10^Ò contains 0.74 pounds of Technical SUMITHRIN^Ò per gallon.
0.74 lbs/gal. X 1 gal./3.785 liters X 453.6 grams/ 1 lb = 88.68 grams/liter
88.68 grams/liter X 1 liter/1000 ml = 0.08868 grams/ml.
There are 0.08868 grams of SUMITHRIN^Ò in each ml of ANVIL 10+10^Ò from the tank.
Convert grams to milligrams. 0.08868 g/ml X 1000 mg/gram = 88.68 mg/ml
PREPARE the STOCK solution by pipetting 1 ml of ANVIL 10+10^Ò into a 100 ml volumetric flask and filling to the 100 ml mark with A.C.S. acetone.
The STOCK solution contains 0.8868 mg SUMITHRIN^Ò/ 1 ml.
Convert milligrams to micrograms. 0.8868 mg/ml X 1000 ug/1 mg = 886.8 ug/ml
We used 1 ml, 2.5 ml, 5 ml and 10 ml of the STOCK solution to prepare TEST solutions.
TEST solutions were prepared by pipetting the STOCK solution into a volumetric flask and filling to the 100 ml mark.
One ml of TEST solution was pipetted into each Wheaton bottle for the bottle bioassay.

RESULTS

There was no difference between the 5 ml and 10 ml dosages, therefore increasing the dosage higher than 10 ml could not kill the mosquitoes any faster.
The 2.5 ml TEST dosage was intermediate between the lowest and highest dosages and gave a desirable mortality curve.
The 2.5 ml TEST dosage is the equivalent of 22.17 ug of SUMITHRIN^Ò per bottle.
2.5 ml STOCK/100 ML x 886.8 ug/ 1 ml = 22.17 ug per bottle.

CONCLUSIONS:

These preliminary results suggest that a dose of 22.17 ug of SUMITHRIN^Ò is a diagnostic dosage for the bottle bioassay.

This information supplements the final concentrations per bottle listed in Table 1 of the CDC protocol.

This information was obtained by testing Ochlerotatus taeniorhynchus field-collected in Pinellas and Monroe Counties. The conclusions stated in this technical memorandum may not apply to other mosquito species or to Oc. taeniorhynchus from other geographic locations.

Baseline data for susceptible Oc. taeniorhynchus are available by CLICKING HERE.

Abbreviations used in this document:

lbs = pounds

g = gram

mg = milligram

ug = microgram

l = liter

gal = gallon

1 gram (g) = 1000 milligrams (mg)

1 milligram (mg) = 1000 micrograms (ug)

Lot number: The research results reported here were obtained using ANVIL 10+10^Ò lot number: 001820-6-1-A.

Acknowledgement:

Three Florida Mosquito Control Districts participated in the work reported here: Citrus County MCD, Pinellas County MCD, and Keys MCD in Monroe County. Thank you to the directors and staff who made this work possible.

ANVIL 10+10^Ò is a registered trademark of Clarke Mosquito Control Products, Inc., Roselle, Illinois 60172.

SUMITHRIN^Ò is a registered trademark of Sumitomo Chemical Company, Ltd., Osaka, Japan
The chemical name of SUMITHRIN^Ò is d-phenothrin.

20 September 2001; revised 5 October 2001; revised 15 October 2001; revised 22 March 2002; revised 1 May 2002; revised 19 April 2004