New Insecticides and Repellents for Use on Mosquitoes and Sand Flies

Ulrich R. BernierUSDA-ARS-CMAVE

Mosquito & Fly Research Unit352-374-5917

uli.bernier@ars.usda.gov

2010 DWFP-OPMRAFebruary 12, 2010Jacksonville NAS

Research Projects

1. Development of Novel Repellents and Insecticides -SCA with Alan Katritzky (UF-Chemistry).

2. Bite Protection of Current Uniforms and Improved Repellent−Treated Military Uniforms - Reimbursable with NSRDEC. Collaboration with Kamal Chauhan (IIBBL), CDR Schoeler (JAX-NECE).

3. Attraction−Inhibitors - Collaboration with Kamal Chauhan (IIBBL), CDR Szumlas, CDR Hoel, LT Furman (NAMRU-3).

Insecticide/Repellent Projects

• 1608 Carboxamides – carboxamide repellents • 1757 Acylpiperidines – best 7 repellents• 1665 Various insecticides – house fly• 1666 Various insecticides – house fly• 1668 Various insecticides – anopheleines• 1669 Various insecticides – Ae. taeniorhynchus• 1670 Various repellents – Ae. aegypti

Project # dataset used from USDA archives

USDA, Agriculture Handbooks No 69 (1954), No 340 (1967), Technical Bulletin No 1549 (1977).

Modeling (ANN, QSAR

Predictions

Synthesis

Bioassays

USDA archives

Research strategy

Gupta RK, Bhattacharjee AK (2007) Insect Repellents: Principles, Methods, and Uses, pp195–228.

Katritzky AR et al. PNAS US. 2008, 21, 7359

“Cloth patch” assay

UF IRB-01 636-2005

Acylpiperidines – top candidates from 1611 repellents (1757)

Complete protection time (CPT) of most active acylpiperidines vs DEET using “Cloth Patch” assays

Co

nce

ntr

atio

n

Pro

tect

ion

time

(day

s)

25 mol/cm2

2.5 mol/cm2

Treatment

Katritzky AR et al. PNAS US. 2008, 21, 7359

Acylpiperidines as mosquito repellents (1757)

Selected carboxamides vs DEETO

N1

O

N3

O

N5

O

N

2

O

N4

N

O

6

N

O

7

N

O

DEET

Carboxamides as mosquito repellents (1608)

Treatment

Co

nce

ntr

atio

n

25 mol/cm2

2.5 mol/cm2

Pro

tect

ion

time

(da

ys)

Complete protection time (CPT) for selected carboxamides and DEET at two concentrations using “Cloth Patch”

assays

N

O

12.0 days

O

N

15.0 days

O

N

13.0 days

O

N

22.0 days

5.0 days

25 and 2.5 mol/cm2

N

O

7.0 days2.5 days

DEET

Carboxamides as mosquito repellents (1608)

Selected carboxamides vs DEET

House Fly, Contact Insecticides (1665)

DatasetPredicted /similarity

#eval

100% Mortality,mmol/L

159 (228)Musca domestica

390/30 83,5-dichloroaniline: 4.6Carbaryl: 14.9Propoxur: 7.1

100 % mortality

NH2Cl

Cl

Con

cent

ratio

n (m

mol

/L)

P/1.5C/3.2

House fly, Residual Insecticides (1666)

Dataset Dataset from archive

Predicted /similarity

#Eval

100% Mortality,mmol/L

53 non-phosph. (out of 228)

Musca domestica 125/25 25

Ethyl cis-3-iodoacrylate: 0.075

Xanthene: 0.005Carbaryl: 0.0149Propoxur: 0.0071

O

100 % mortality

P/1.4 C/3

Con

cent

ratio

n (m

mol

/L)

0

0.005

0.01

0.015

0.02

Carbaryl Propoxur

Common Malaria MosquitoResidual Insecticides (1668)

ON

O

F

O N

O

Cl

Dataset from archivePredicted/similarity

#eval

100% Mortality,mmol/L

62 non-phosph. (241) Anopheles

230 28

2-(3-fluorophenoxy)-benzoxazole: 0.0130882-(4-chlorophenoxy)-benzoxazole: 0.006106

Carbaryl: 0.0149Propoxur: 0.0071

100 % mortality

P/1.2C/2.5

Con

cent

ratio

n (m

mol

/L)

Evaluation of Permethrin-Treated Military Uniforms

• August, 2003—After Monrovia, Liberia mission, USMC requested assistance of MFRU to transition to factory-treated uniforms.

• May 20, 2006-March 09, 2007—”First Article Testing” of Marine Corps Combat Utility Uniforms (MCCUUs).

• 2008-2009—Evaluation of U.S. Army, USMC, U.S. Air Force, U.S. Navy uniforms.

• 2010—”First Article Testing” of Factory Treated Fire-Resistant Army Combat Uniforms (FT FRACUs).

“Bite Protection” AssayUF IRB-01 69-2006

Specimen(Co)-Type-Treatment

Air Perm(cfm)

Bite Protection (%±SE)

0x 20/25*x 50x

AA-MCCUU-Trouser 6.8 99.1 ± 0.3 97.5 ± 0.7 93.7 ± 1.0EA-MCCUU-Trouser 6.8 98.4 ± 0.5 99.2 ± 0.4 96.0 ± 0.8CO-MCCUU-Trouser 6.8 96.0 ± 0.7 98.3 ± 0.6 97.6 ± 0.6PR-MCCUU-Trouser 6.8 95.9 ± 0.9 96.8 ± 0.7 90.3 ± 1.7MCCUU-Tr WK Rev Ord 6.8 90.8 ± 2.0 n/a n/aAA-MCCUU- Blouse 10.4 99.1 ± 0.2 98.7 ± 0.3 97.9 ± 0.5EA-MCCUU- Blouse 10.4 98.5 ± 0.4 97.8 ± 0.6 97.5 ± 0.7CO-MCCUU- Blouse 10.4 97.7 ± 0.3 98.7 ± 0.2 98.5 ± 0.5PR-MCCUU- Blouse 10.4 97.4 ± 0.7 97.5 ± 0.6 96.4 ± 0.8ABU IDA* 10.8 94.5 ± 1.5 92.3 ± 1.6 82.8 ± 3.0Abrams V 23.0 97.0 ± 0.4 97.4 ± 0.4 97.4 ± 0.5Abrams Lite iCVC 37.5 95.6 ± 0.7 94.5 ± 0.9 84.9 ± 3.3FROG Sleeve 44.1 92.1 ± 1.4 92.9 ± 1.1 87.6 ± 1.8FROG Torso 114.0 71.3 ± 3.6 49.9 ± 4.6 49.3 ± 4.1

Summary of Bite Protection Evaluation of Twill Uniforms

(MCCUUs, FROG, ABUs, Abrams)

Summary of Bite Protection Evaluation of US Army Uniforms:

ACUs, FRACUs, Nomex

Specimen(Co)-Type-Treatment

Air Perm(cfm)

Bite Protection (%±SE)

0x 20/25*x 50x

ACU IDA* 6.1 99.9 ± 0.1 98.8 ± 0.5 95.1 ± 0.9

BI-ACU-FT 6.1 98.9 ± 0.4 99.5 ± 0.3 tbd

FRACU IDA 55.9 91.2 ± 0.9 n/a n/a

WK-FT-FRACU 55.9 95.9 ± 0.5 87.4 ± 1.9 74.9 ± 2.6

BO-FT-FRACU 55.9 92.1 ± 1.0 92.6 ± 1.2 76.3 ± 1.9

WK-S-FRACU Reform Bind 55.9 86.1 ± 1.2 83.8 ± 0.9 80.9 ± 1.2

BO-R2-FRACU Reform Bind 55.9 91.8 ± 0.9 87.2 ± 1.9 78.3 ± 2.5

Nomex iCVC/A2CU 96.5 89.0 ± 1.7 58.6 ± 3.4 48.9 ± 3.4

Comparison of Air Permeability (CFM) and“Bite Through” (%) of Twill and Ripstop Combat

Uniforms

0

10

20

30

40

50

60

70

80

90

100

MCCUTrouser

MCCUUBlouse

Abrams V Abrams Lite ACU S/7327 S/7326 S/7328 S/7320 DEF M

Bit

e T

hro

ug

h (

%)

Air Permeability %Bite Through

Air permeability (cfm)TwillRipstop

U

Evaluation of NWU Field Treatment:Hand Can, Spray Can, IDA Kit

(Collaboration with CDR Schoeler,JAX-NECE)

Bite protection evaluation initiated 18 Nov 2009-Initial (0x laundered specimens)

3 volunteers [control bite through 30.5 ± 7.7 %]

Hand Can (BP%) Spray Can (BP%) IDA Kit (BP%)

Ae. aegypti 100.0± 0.0 88.0 ± 3.7 98.8 ± 1.2

An. albimanus 96.6± 2.0 90.8 ± 1.0 100.0 ± 0.0

Treated Fabric Strips

• FRACU fabric obtained from Natick Soldier Center

• Single-ply fabric cut into (25.5 cm x 33.2 cm x 21.0 cm side) trapezoids (691 cm2 area) and sewn into sleeves.

• Chemicals: Permethrin on uniform plus additional fast-acting insecticides ( with Kamal Chauhan-IIBBL).

http://www.16mmdrive-infilms.com/pb/wp_66114d41/images/img296954970d2e00fb40.jpg

Evaluation of untreated and permethrin treated sleeves with a (20.5 cm x 2.6 cm) untreated, several volatile pyrethroids, or volatile

insectide-treated fabric strips affixed with Velcro® to the wrist regionMaterial was the US Army Fire Resistant Army Combat Uniform (FRACU)

(65% Rayon, 10% Nylon, 25% Para-aramid)

Treated FRACU (0.134 mg/cm2)/treated strip (0.158 mg/cm2)

DEET-protected hand

Absence of odour sensitivity

Absence/decrease of sense of smell

Lack of or decrease in olfaction, either permanent or temporary

On average used ~2 times of 100 million spoken words in English (Websters Online)

Anosmia (noun)

A typical repellent functions as a feeding deterrent or contact irritant. The surface is

unappealing to land on, remain in contact with, or feed through.

Adapted from: Bernier, U.R., Laboratory Research and Development of Attractants, Inhibitors and Repellents, Tech. Bull. Florida Mosq. Control Assoc., 2006, 9:9-16.

An attraction-inhibitor decreases host-finding by masking or cloaking the presence of kairomones – evident by anosmia exhibited by the insects.

?

Adapted from: Bernier, U.R., Laboratory Research and Development of Attractants, Inhibitors and Repellents, Tech. Bull. Florida Mosq. Control Assoc., 2006, 9:9-16.

Non-competitive assays: Addition of a small amount of inhibitor to an attractant blend or to human emanations

results in decreased attraction of Ae. aegypti

92.7

12.8

88.6

23.5

0

10

20

30

40

50

60

70

80

90

100

Att

ract

ion

(%

)

Blend Blend + I7 Hand Hand + I7Treatment

Data acquired 20-26 June 2000

Non-competitive assays: Candidate attraction-inhibitors combined with the

attractant blend (Aedes aegypti) Tukey’s HSD (P = 0.05) (F45,287 = 59.6, P < 0.001)

0%10%20%30%40%50%60%70%80%90%

100%

I1 I2 I3 I4 I5 I6 I7 I8 I9 I10

I11

I12

I13

I14

I15

I16

I17

I18

I19

I20

I21

I22

I23

I24

I25

I26

I27

I28

I29

I30

I31

I32

I33

I34

I35

ATTR I3

6I3

7I3

8I3

9I4

0I4

1I4

2I4

3I4

4I4

5

Treatment

Att

ract

ion

(%

)

Efficient Inhibition

Data acquired 2 May 2005 – 23 July 2007

ab

c

de

31.9%

Non-competitive assays: Response of Anopheles albimanus to a subset of

attraction-inhibitorsAttractant = L-lactic acid + methylene chloride

Tukey’s HSD (P = 0.05) (F14,95 = 15.2, P < 0.001)

0%10%20%30%40%50%60%70%80%90%

100%

I9 I8 I15 I3 I10

I21 I7 I2 I12 I1 I16

I13

I45

I24

ATT

R

Treatment

Att

ract

ion

(%

)

Efficient Inhibition

a

bc

d

Data acquired 9 May 2006 -11 September 2007

Evaluation of Selected Inhibitors in a Single-Tube Two-Port Olfactometer

Phlebotomus papatasi

Evaluation of Inhibitors with MM-X Traps in Aswan, EgyptCollaboration with NAMRU-3> 93% Phlebotomus papatasi

Release devices – designed for> 24 h protection

Comparison-2007 vs. 2006 total sand flies

Over 12 Trap Nights

CO2 GroundCO2 Up

CO2 + DBSCO2+Oct/NH3

Hair + CO2Octenol + CO2

CO2 onlyUnbaited

0

250

500

750

1000

1250

1500

1750

2000

2250

2500

Treatment

San

d F

lies

Cau

gh

t (N

um

ber

) 2006

2007

Technical StaffPhotos

Greg AllenJesse DurranceNatasha ElejaldeNathan NewlonMaia Tsikolia

Dan Kline, Jerry Hogsette--Honorary Members

Probability that I would not mention Dan Kline in a talk: